PMID- 6269944 TI - Dietary fiber and personality factors as determinants of stool output. AB - We examined effects of fiber on stool output, since this is one of the primary mediating variables for the hypothesized relationship between fiber and disease. Total neutral detergent fiber in the dietary fiber source was predictive of stool weight but not frequency. Substantial individual differences in stool output remained when dietary factors were controlled. Personality measures were used to predict stool weight and frequency independently of diet, and accounted for about as much variance in stool output as did dietary fiber. These results suggest that personality factors predispose some persons to low stool output. These individuals may benefit particularly from dietary fiber. PMID- 6269945 TI - Effect of lithium ingestion on water and electrolyte transport in rat intestine. AB - The effect of lithium ingestion on intestinal electrolyte and water transport was studied in adult Sprague-Dawley rats. We fed animals a lithium-supplemented diet for 1, 2, 4, or 16 wk before in vivo perfusion of the jejunum and colon. Lithium feeding did not alter jejunal transport of water, electrolytes, or glucose, However, at 4 and 16 wk (16-wk data given) the colon increased net water (168%), sodium (160%), and chloride (140%) absorptions, and the transmural potential difference (396%) as compared with control animals. In addition, the colon absorbed both bicarbonate and potassium against an unfavorable electrochemical gradient. The increased colonic sodium absorption was not associated with an increase in mucosal Na+, K+-ATPase activity. Furthermore, in lithium-fed animals deoxycorticosterone acetate stimulated mucosal Na+, K+-ATPase activity, but it did not further increase net sodium absorption. Neither jejunal nor colonic electrolyte transport was affected 24 h after being gavage-fed lithium. These results suggest that chronic lithium ingestion has a unique mechanism of action as other means of chronically increasing sodium absorption are associated with increased mucosal Na+, K+-ATPase activity. PMID- 6269946 TI - Inhibition of gastric emptying in humans by secretion, the octapeptide of cholecystokinin, and intraduodenal fat. PMID- 6269947 TI - High plasma HCO3-protects gastric mucosa against acute ulceration in the rat. PMID- 6269948 TI - Alteration of the granulomatous response in murine schistosomiasis by the chronic administration of captopril, an inhibitor of angiotensin-converting enzyme. AB - Captopril (SQ 14225) is an inhibitor of angiotensin I-converting enzyme (ACE). Mice with schistosomiasis form granulomas in the liver and intestines that have high ACE activity. We determined whether chronic oral captopril administration would alter the granulomatous response. Infected mice treated with captopril had a dose-dependent decrease in liver granuloma size. Chronic peroral administration of drug induced a sustained decrease in granuloma ACE activity and size which was reversible upon drug withdrawal. The drug was most effective during the acute phase of infection. Granuloma size in the colon, ileum, and ileal Peyer's patches was also decreased by treatment. Ileal granulomas were affected the least. Liver, colon, ileum, and ileal Peyer's patches demonstrated natural differences in size of response to schistosome eggs. Synchronous hypersensitivity granulomas, containing no ACE activity, were generated by the pulmonary embolization of methylated bovine serum albumin-coated, Sepharose beads into sensitized mice. Captopril treatment did not affect the size of these lesions. Sustained improvements in portal pressure, body and liver weight, and water consumption were seen in treated mice. We conclude that captopril induces sustained suppression of the granulomatous response in schistosome-infected mice, which results in decreased morbidity. Circumstantial evidence suggests that captopril influences inflammation through the competitive inhibition of ACE. PMID- 6269950 TI - Ligand-binding and autoradiographic studies of GABA-receptors in tissue culture. PMID- 6269949 TI - Prevention of cholesterol gallstones by lignin and lactulose in the hamster. AB - The effect on prevention of cholesterol gallstones by a nonfermentable type of fiber, lignin, and a fermentable fiber analogue, lactulose, was studied in hamsters fed an essential fatty acid deficient diet. Control animals had a high incidence of cholesterol gallstones (21 of 24) and lithogenic bile (lithogenic index 1.08). Animals fed lignin had significantly fewer gallstones (11 of 25), improved cholesterol saturation of gallbladder bile, and increased fecal bile acid excretion. Lactulose-fed animals had significantly fewer gallstones (12 of 24) but no significant change in cholesterol saturation of gallbladder bile or in fecal bile acid excretion. Serum cholesterol concentration was reduced, however, and fecal neutral steroid excretion was increased. Gallstones were completely prevented in animals fed both lignin and lactulose (0 of 22), but gallbladder bile cholesterol saturation was not significantly different from the lignin-fed group. Gallbladder bile mucopolysaccharide concentrations did not differ among groups. Lignin appears to prevent cholesterol gallstones in this model by improving cholesterol saturation of bile. The mechanism of action of lactulose is not yet clear. PMID- 6269951 TI - Cannabichromene and delta 9-tetrahydrocannabinol: interactions relative to lethality, hypothermia and hexobarbital hypnosis. PMID- 6269952 TI - The immaturity of interactions between GABA- and benzodiazepine binding sites in the frog spinal cord. PMID- 6269953 TI - Characterization of NaK-ATPase from vascular smooth muscle. PMID- 6269954 TI - Plasma concentration of corticosterone during Rana catesbeiana tadpole metamorphosis. PMID- 6269955 TI - The mode of action of the crustacean neurosecretory hyperglycemic hormone. I. Involvement of cyclic nucleotides. PMID- 6269956 TI - Occurrence and binding affinity of prolactin receptors in amphibian tissues. PMID- 6269957 TI - Aminergic involvement in the control of luteinizing hormone secretion in the domestic fowl. PMID- 6269958 TI - The nucleotide sequence and protein-coding capability of the transposable element IS5. AB - The nucleotide sequence of IS5, a bacterial insertion sequence, has been determined. It is 1195 bp long and contains an inverted terminal repetition of 16 bp with one mismatch. One open reading frame, spanning nearly the entire length of the element, could encode a polypeptide of 338 amino acids. Upon insertion into a DNA segment, IS5 causes a duplication of 4 bp. Based on seven examples, this site of insertion appears to be nonrandom, and the consensus target site sequence is C . T/A . A . G/A (or C/T . T . A/T . G on the opposite strand). The nucleotide sequences of IS5 insertions into the B and cim genes of bacteriophage Mu have allowed tentative identification of the protein-coding frames of B and cim. PMID- 6269960 TI - Restriction mapping of the DNA of the Streptomyces temperate phage phi C31 and its derivatives. AB - DNA of phi C31 propagated on Streptomyces lividans 66 contained no sites for the restriction enzymes BamHI, SalPI (=PstI) and XhoI; one for XbaI; three for HpaI; five for ClaI and KpnI; six for EcoRI; about 13 for HindIII; about 14 for BclI; and more than 15 for FspAI, HgiAI, SacI, SalGI and SmaI. A complete map of 20 sites (XbaI, HapI, ClaI, KpnI and EcoRI) was obtained using partial digestion and double digestion of DNA of the wild-type and deletion and insertion mutants. The total molecular size was estimated to be 41.2 kb. PMID- 6269959 TI - The nucleotide sequence of IS5 from Escherichia coli. AB - A 3-kb fragment of Haemophilus haemolyticus DNA which carries the HhaII restriction (r) and modification (m) genes has been cloned into the PstI site of pBR322 (Mann et al., 1978). When propagated in Escherichia coli, it was observed that spontaneous insertions of IS5 inactivated the restriction gene, producing r- mutants at a frequency of 10(-6). Electron microscopy, restriction-site mapping and sequence analysis of two r- plasmids have demonstrated the presence of IS5 at a single target site in both possible orientations. The complete nucleotide sequence of IS5 has been determined. It is 1195 bp long and has inverted terminal repeats of 16 bp. The target site for IS5 in this plasmid is 5'-CTAG. Approx. ten copies of IS5 were found to be present at about the same locations on the E. coli chromosome in various K-12 strains, using Southern hybridization analysis. PMID- 6269961 TI - Physical and genetic organization of the IncN-group plasmid pCU1. AB - A restriction endonuclease-cleavage map of the IncN group plasmid pCU1 was constructed. Deletion mutants of the plasmid were obtained by in vivo or in vitro methods. Comparison of the restriction maps of these mutants to that of pCU1 enables one to assign the known functions of the plasmid to particular regions on the plasmid DNA. For different enzymes, the number and distribution of restriction sites on pCU1 is compared to that of other IncN and related plasmids. PMID- 6269962 TI - Cloned genomic segments of Zea mays homologous to zein mRNAs. PMID- 6269963 TI - Transposition of a DNA fragment flanked by two inverted Tn1 sequences. AB - The 32 Md fragment (derived from plasmid RP4::Tn1) carrying the Kmr gene and flanked by two inverted Tn1 elements is capable of recA-independent translocation to other plasmids. We designated this new transposon Tn1755. In various crosses, frequencies of Tn1755 transposition to plasmids Co1B-R3, R15 and F'Co1VBtrp varied from 2.5 to 90% of the frequencies of Tn1 transposition. Tn1755 can integrate into various sites of the recipient plasmids. We failed to observe transposition of another RP4::Tn1 fragment flanked by two opposingly oriented Tn1 transposons and harboring the Tcr gene. Presumably, to form a new transposable structure, other features must also be of importance. PMID- 6269964 TI - [Differential diagnosis of peripheral vasomotor disorders of vibration and nonvibration origin]. PMID- 6269965 TI - [Health status of workers in the manufacture of artificial diamonds and diamond tools]. PMID- 6269966 TI - [Determination of free silicon dioxide in the dust from cement manufacture]. PMID- 6269967 TI - [Participation of cyclic AMP in the mechanism of the effect of LH on the steroidogenesis in the ovaries]. PMID- 6269968 TI - Loperamide: studies on its mechanism of action. AB - The effects of loperamide on net solute and water absorption, and prostaglandin E2 (PGE2) and cholera toxin-induced secretion were studied in the rat jejunum using an in vivo steady-state perfusion technique. Loperamide stimulated absorption of fluid, electrolytes, and glucose and reversed PGE2 and cholera toxin-induced secretion to absorption; this opiate analogue had no effect on cholera toxin stimulation of adenylate cyclase activity or the rise of tissue cyclic AMP (cAMP) concentrations. The opiate antagonist, naloxone, reduced the antisecretory effects of loperamide without affecting tissue levels of cAMP. These results indicate that loperamide inhibits PGE2 and cholera toxin-induced secretion, and that this phenomenon is independent of any direct effect that cholera toxin has on the adenylate cyclase system. The action of naloxone suggests, but does not prove, that loperamide exerts its effect via opiate receptors. PMID- 6269969 TI - A review of enzyme changes in serum and urine due to treatment with drugs (tuberculostatics, contraceptive medication, diagnostics and drugs in real diseases). PMID- 6269970 TI - [Effect of lithium carbonate on the function and enzyme content of the neutrophils in patients with granulocytopenia]. PMID- 6269971 TI - [Pharmacological studies on experimental nephritic rats (13). The development and aggravation of nephritis due to the repeated administration of liquoid (author's transl)]. AB - In order to elucidate the role of blood coagulation system in the development and aggravation of glomerulonephritis, liquoid (Liq) was repeatedly administered to normal or nephritic rats. When Liq 10 mg/kg was given i.v. daily X 22 to normal rats (group I), the urinary excretions of protein and N-acetyl-beta glucosaminidase and urea nitrogen content did not significantly change as compared with those in the normal control group. In rats given Liq 10 mg/kg i.v. either every 3 days X 8 (group III) or every day X 22 (group IV) from the 15th day after the i.v. administration of anti-rat glomerular basement membrane rabbit serum (AGS) [0.5 ml/150g body weight], these biochemical parameters were not significantly different from those of nephritic control rats given AGS only (group II). The deposits of fibrin or fibrinoids in glomeruli of groups I, II, III and IV, examined by a fluorescence antibody technique were evident in 2, 2, 8 and 10, respectively, out of 10 rats in each group, although the degree of deposition was slight. Under light microscopy, the adhesion between the glomerular capillary wall and Bowman's capsule, hypercellularity, crescent formation and hyalinization were demonstrated in a part of glomeruli, even in group I. concerning the influence of Liq in nephritic rats, the most prominent glomerular change was hyalinization. While in group II the hyalinization was evident in only 17% of glomeruli, in groups III and IV the hyalinization was 41 and 55%, respectively. Although no significant difference was seen between groups II and III regarding other glomerular changes except for hypercellularity, these changes in group IV increased as compared with those in group II. However, hypercellularity was less in groups III and IV than in group II. A slight occlusion of the glomerular capillary lumen was observed, even in group I. In nephritic groups, the degree of the capillary lumen occlusion in group IV was greater than that in group II. From these results, the acceleration of intraglomerular blood coagulation is considered to be a major factor in the development and aggravation of glomerulonephritis. PMID- 6269972 TI - Catabolite repression of different inducible enzymes in Escherichia coli and the effect of cAMP. AB - Simultaneous induction of two enzymes sensitive to catabolite repression does not lead to an additive decrease of the specific activity of the two. Exogenously added cAMP increases the specific activity of catabolically repressed enzymes, irrespective of whether the enzyme is induced separately or simultaneously with another enzyme. In the presence of 12 different substrates metabolized by inducible enzymes glucose does not bring about catabolite repression. Synthesis of cAMP is identical with that occurring under conditions when glucose brings about catabolite repression. PMID- 6269973 TI - Presence of ACTH-potentiating factors in rat anterior pituitary glands. AB - High molecular weight ACTH fractions, obtained through gel filtration of boiled rat anterior pituitary extract, induced a marked increase in corticosterone production from isolated rat adrenal cells in the presence of low concentrations of ACTH-(1-24). This indicates the presence of heat-stable factors augmenting the steroidogenic action of ACTH in the rat anterior pituitary. We also noted that these factors potentiated the activity not only of ACTH-1(1-24) but also of ACTH (1-8). The ACTH-potentiating factors in rat anterior pituitary extract are possibly present in heterogeneous forms according to their molecular weights (8,000, 10,000 and 15,000), their mobility in ion-exchange chromatography and their content in RIA-ACTH activity. Of these three forms, the former comigrated with biological ACTH activity. The remaining two forms were free of it. Since the effect of potentiating factors on modified ACTH-(1-9), shown to be less susceptible to proteolytic degradation from ACTH-(1-24), was similar to the effect on ACTH-(1-24), it is suggested that potentiation was not due to an inhibition of ACTH proteolysis. PMID- 6269974 TI - Identification of integrated hepatitis B virus DNA sequences in human hepatocellular carcinomas. AB - DNA extracts from hepatocellular carcinomas of 13 patients from South Africa were examined for hepatitis B virus (HBV) DNA sequences by molecular hybridization using [32P]-labeled recombinant, cloned, and purified HBV-DNA. Eight patients were HBV carriers as demonstrated by the presence of hepatitis B surface antigen (HBsAg) in their serum, and each of these patients and HBV-DNA sequences in hepatocellular carcinoma tissue. Five patients who were not HBsAg carriers, did not have HBV-DNA in their tumors. In DNA extracts from all tumors of patients who were HBsAg-positive, the HBV-DNA was integrated into the host genome. The integration pattern was unique for each tumor, but HBV-DNA bands of a given length were present in more than one specimen and in a human hepatocellular carcinoma cell line (PLC/PRF/5). These results suggest that integration of HBV DNA into the human genome occurs in conjunction with malignant transformation. PMID- 6269975 TI - Hepatitis B virus and hepatocellular carcinoma: molecular biology provides further evidence for an etiologic association. PMID- 6269976 TI - The Mallory body: structure, composition, and pathogenesis. PMID- 6269977 TI - Transferrin receptors and the uptake and release of iron by isolated hepatocytes. AB - Isolated rat hepatocytes accumulate iron from iron-transferrin by a process which is dependent on the temperature and on the transferrin concentration, and which is diminished by treatment of the cells with a proteolytic enzyme. These observations are consistent with a mechanism for iron uptake into hepatocytes involving the binding of iron-transferrin to a specific cell-surface receptor. Apotransferrin is also able to bind to the hepatocyte but the apparent binding constant is about 35 times lower than that observed for the binding of iron transferrin. The binding of apotransferrin to the cells is completely abolished by a low concentration of iron-transferrin. This suggests that the apotransferrin is binding weakly to the same receptor to which iron-transferrin binds and that there are not receptors on the surface of the hepatocyte specific for apotransferrin. In the absence of such specific-binding sites, apotransferrin may act as a passive acceptor of iron released from the hepatocyte. PMID- 6269978 TI - Presence of receptors for polymerized albumin in HBsAg-containing hepatocytes and hepatoma cell line. AB - Recent evidence suggests that hepatitis B virions (HBV) and HBsAg particles contain receptors for polymerized human serum albumin (pHSA). We studied, by immunohistochemical techniques, the relationship between HBsAg and pHSA receptors in liver tissue from eight patients with chronic HBV infection and in a human hepatocellular carcinoma cell line (PLC/PRF/5) producing HBsAg. Both parallel sections and double fluorescent antibody staining of liver tissue demonstrated that only HBsAg-containing hepatocytes expressed pHSA receptors. The receptors could not be demonstrated in eight HBsAg negative livers. Sequential studies of PLC/PRF/5 cells revealed that pHSA and HBsAg emerged simultaneously in the cytoplasm, on the cell surface, and in the supernatant culture media. These findings indicate that pHSA receptors are closely associated with HBsAg during its synthesis and secretion by hepatocytes and suggest that the receptors are HBV specific. PMID- 6269979 TI - In vitro effect of bile salts on rat liver plasma membrane, lipid fluidity, and ATPase activity. PMID- 6269980 TI - Regulation of the metabolism of plasma membrane proteins in hepatocytes. PMID- 6269981 TI - Woodchuck hepatitis and hepatocellular carcinoma: correlation of histologic with virologic observations. AB - The livers of 33 captive woodchucks were examined histologically in 30 biopsy and 10 autopsy specimens and the findings were correlated with serum determinations for woodchuck hepatitis virus (WHV), surface antigen (WHsAg) and antibody (anti WHs), and WHV DNA and DNA polymerase. The liver appeared normal in all 3 serum negative animals, 7 of 16 with indeterminate WHV status, and 1 of 4 with anti WHs, but not in 10 animals with WHsAg, WHV DNA, and DNA polymerase. Mild hepatic inflammation was found in 7 woodchucks with indeterminate status, 4 with anti WHs, and 2 with each marker of WHV infection. Significant inflammation was found in 2 of indeterminate status and 4 with every marker, whereas more severe lesions (2 of chronic active type) occurred, almost always in autopsy specimens, in 8 animals with every marker. Eight of 10 animals with all markers had orcein positive inclusions (Shikata's technique) and 6 had hepatocellular carcinoma associated with acute and chronic hepatic inflammation and, usually, neoplastic nodules in the noncarcinomatous parenchyma. Features distinguishing the woodchuck lesion from human hepatitis B disease were: association of carcinoma with acute hepatic inflammation (but not with cirrhosis) and DNA polymerase in the serum; transition to carcinoma from neoplastic nodules; conspicuous plasma-cellular reaction of hepatic inflammation, and hematopoietic cells in the tumor. Significant hepatic lesions in the woodchucks were regularly associated with serum WHsAg, WHV DNA, and DNA polymerase. In contrast to man, hepatocellular carcinoma in woodchucks was regularly associated with these markers of active viral replication. The nature of the orcein-positive inclusions requires elucidation, although they may assist in screening for similar viruses in other species. The woodchuck may help in the study of the relation between hepatocellular carcinoma and hepatitis B, including the possibility of cocarcinogenic factors. PMID- 6269982 TI - Granuloma collagenase and EDTA-sensitive neutral protease production in hepatic murine schistosomiasis. AB - Mice infected with Schistosoma mansoni represent a model for study of hepatic fibrosis in humans. Production of trypsin-activatable inactive collagenase and EDTA-sensitive neutral protease was measured in the culture medium in which granuloma explants or primary cultures were maintained. Collagenase production was maximal in granulomas obtained from liver of mice 8 weeks postinfection and was inhibited by Actinomycin D or cycloheximide, and enhanced by lymphocyte factor(s) or heparin. Isolated schistosome eggs did not release these enzymatic activities but did release EDTA-insensitive protease activity. Both enzymes were separated by ion-exchange chromatography and purified to homogeneity. Isolated collagenase had an isoelectric point of 6.2 and molecular weight of 60,000 and had the functional characteristics of a tissue collagenase. The specific activity of collagenase was 33 units per mg protein at an optimum pH 7.5 and lacked proteolytic activity against noncollagenous protein substrates. Isolated EDTA sensitive neutral protease had specific caseinolytic activity of 150 units per mg protein and gelatinolytic activity of 300 units per mg protein at an optimum pH 7.5; the enzyme lacked activity against undenatured collagen. Isoelectric point was pH 6.0. Protease activity was inhibited by known inhibitors of collagenases. Production and activation of EDTA-sensitive neutral protease and collagenase accompany increased collagen synthesis and content in the liver of mice 8 weeks postinfection with S. mansoni cercariae. Continued accumulation of liver collagen under these conditions suggests an insufficiency in collagenase activity relative to the increase in collagen synthesis. PMID- 6269984 TI - Hepatocyte bile secretion: current views and controversies. PMID- 6269983 TI - Ligandinemia in primary liver cell cancer in rat and man. AB - Ligandin was quantitated by radioimmunoassay in serum and, when possible, in tumors from patients with primary hepatocellular carcinoma, massive hepatic metastasis or nonhepatic primary neoplasms, and in rats and athymic (nu/nu) mice bearing transplantable ligandin-containing or nonligandin-containing rat hepatocellular carcinomas. Following transplantation of a ligandin-containing rat hepatocellular carcinoma in rats or athymic (nu/nu) mice, mean serum ligandin concentrations progressively increased and, within 4 months, exceeded normal serum ligandin concentrations by 10-fold. In 11 of 15 HBsAg negative patients with primary hepatocellular carcinoma, serum ligandin concentrations ranged from 82 to 551 (mean: 298) ng per ml; the mean ligandin concentration in the hepatic neoplasm was 32% of the ligandin concentration in normal liver. In 19 of 22 patients with extensive hepatic metastasis and in each of 20 patients with primary carcinomas without hepatic metastasis, serum ligandin concentrations were normal (7.0 +/- 4.0 ng per ml). PMID- 6269985 TI - A dilutional immunoperoxidase study of proliferative ductal lesions and carcinomata of the breast. AB - A dilutional immunoperoxidase study of carcinoembryonic antigen (CEA) reactivity of mild and severe epitheliosis as well as malignant ductal and lobular lesions of the breast was performed. Intraduct carcinoma with a cribriform and clinging patterns showed intracytoplasmic, glycocalyceal and intraluminal staining for CEA at higher dilutions of antiserum than cases with mild and severe epitheliosis. Also, many intraduct carcinomas associated with infiltration stain at a lower concentration of antiserum to CEA than pure intraduct lesions. Staining is seen at lowest concentrations in the infiltrating components. This suggests that as intraduct carcinoma becomes invasive, it loses some of its ability to store and possibly to synthesize CEA. The dilutional immunoperoxidase method could be applied routinely to cases of severe epitheliosis to differentiate them from intraduct carcinoma. However, relatively few cases were studied in this preliminary series and a further study is being carried out. The difficulties encountered using CEA as a tumour marker are outlined. PMID- 6269986 TI - A comparative study of referral success in first appointments made by patients and by staff. PMID- 6269987 TI - Epstein-Barr virus and human disease: immune responses determine the clinical and pathologic expression. PMID- 6269988 TI - The biochemical basis for genotyping 21-hydroxylase deficiency. AB - We describe three different forms of 21-hydroxylase deficiency-classical congenital adrenal hyperplasia (CAH), late-onset 21-hydroxylase deficiency, and cryptic 21-hydroxylase deficiency-and we present hormonal standards by which to assign the appropriate 21-hydroxylase deficiency genotype for these disorders. The late-onset and cryptic forms of 21-hydroxylase deficiency are biochemically indistinguishable, although patients with the late-onset disorder present with marked clinical symptoms (e.g. virilization) whereas patients with cryptic 21 hydroxylase deficiency are clinically asymptomatic. Our latest studies suggest that late-onset 21-hydroxylase deficiency, like the classical and cryptic 21 hydroxylase deficiencies, is also genetically linked to HLA, the major histocompatibility complex of man. Our biochemical findings provide evidence that a spectrum of 21-hydroxylase deficiencies exist in the population. PMID- 6269990 TI - Cell requirements for optimum proliferative responses of rat spleen cells. PMID- 6269989 TI - Fanconi's anemia: anomaly of enzyme passage through the nuclear membrane? Anomalous intracellular distribution of topoisomerase activity in placental extracts in a case of Fanconi's anemia. AB - In cells of Fanconi's anemia (FA) spontaneous breakage of chromosomes was first recognized by Schroeder et al. (1964). Sensitivity to bivalent alkylants has been found to be a constant feature, whereas low levels of several repair-related enzymes have been described in different FA cell lines. In a family with known FA, during a further pregnancy the prenatal diagnosis of the disease was made by cytogenetic analysis of amniotic cells. After birth the fresh placenta was extracted for further enzymologic analysis. An unusual distribution of DNA topoisomerase activity was noted: high in the cytoplasm and only a little activity in the nuclear sap. This contrasts with findings in normal placentae. Since amniotic cells, lymphocytes, and fibroblasts of this child exhibited both high spontaneous breakage of chromosomes and sensitivity to the bivalent alkylant, diepoxybutane, a correlation between the findings on cytogenetic and enzymologic levels is assumed. Whereas in other published cases, a true reduction of activities of enzymes involved in DNA replication and repair has been found, the present results suggest the interpretation that in our patient the genetic anomaly does not affect the level of synthesis of the enzyme itself, but the passage of the enzyme from the place of synthesis (the cytoplasm) to the substrate (inside the nucleus). A genetic anomaly of the nuclear membrane might be a possible explanation, or alternatively, a structural mutation of the enzyme at a site not affecting the catalytic activity, but affecting the membrane passage or intranuclear accumulation. Meanwhile, placentae of two other cases gave similar results, thus supporting our findings. PMID- 6269992 TI - Comparison of extracellular & intracellular level of cyclic AMP between the toxinogenic & non-toxinogenic strains of Vibrio cholerae. PMID- 6269991 TI - Chronic effects of drugs on liver histology & serum aminotransferases in rabbits maintained on high fat diet. PMID- 6269993 TI - Bilateral cryptophthalmos-syndactyly syndrome. PMID- 6269994 TI - Herpes zoster ophthalmicus in children (reports of two cases). PMID- 6269995 TI - Solid malignant tumours of infancy and childhood--a clinico-pathological study. PMID- 6269996 TI - Antibody response to three doses of conventional and double-dose OPV and to six doses of conventional OPV. PMID- 6269997 TI - Interaction of cytomegalovirus immune complexes with host cells. AB - After adsorption to host cells of a mixture of cytomegalovirus (CMV) and immunoglobulin G (IgG)-anti CMV, at least 99% of the surviving fraction consisted of infectious virus-antibody complexes which could be neutralized by anti-human IgG antibody. The virus-antibody complexes penetrated into cells and were uncoated more slowly than native virus. When the virus-antibody complexes were treated with anti-human IgG antibody before adsorption was allowed to take place, anti-human IgG antibody immune complexes were adsorbed to host cells less readily and uncoated less efficiently than were untreated complexes. Furthermore, after adsorption, anti-human IgG antibody-IgG-CMV complexes were further retarded in their penetration and uncoating. These observations suggest that neutralization of CMV by IgG-anti-CMV resulted from interference with the normal mechanisms of penetration and uncoating. Anti-human IgG antibody enhanced the degree of neutralization by augmenting the inhibitory effect of antibody on these stages of virus-host interaction and also reduced adsorption of the complex. PMID- 6269998 TI - Recovery of mice from herpes simplex virus type 2 hepatitis: adoptive transfer of recovery with immune spleen cells. AB - Young BALB/c mice inoculated intraperitoneally with herpes simplex virus type 2 develop focal necrotizing hepatitis. After infection, the livers of these mice show increasing virus titers, which reach a maximum on day 3 after infection; this is followed by a dramatic decrease in the amount of virus recovered on days 4 and 5. This decrease in virus content is accompanied by a progressive infiltration of the lesions with mononuclear leukocytes and an apparent resolution of the lesions. Adoptive transfer of immune spleen cells from mice infected 6 days earlier accelerated this process. When 50 x 10(6) to 100 x 10(6) immune spleen cells were transferred 24 h after infection, the inflammatory response and the clearance of virus from the livers were advanced by almost 2 days. As few as 12 x 10(6) immune spleen cells accelerated the healing process, whereas fewer immune cells, disrupted immune cells, or normal spleen cells did not have an effect. The protection conferred by herpes simplex virus type 2 sensitized immune spleen cells was specific since mouse cytomegalovirus- or vaccinia virus-sensitized immune spleen cells had no effect on the course of infection with herpes simplex virus type 2, whereas some cross-reactivity was observed between herpes simplex virus types 1 and 2. This model seems to be suitable for examining the immunological mechanisms that are active during recovery from visceral herpes simplex virus infections. PMID- 6269999 TI - Biological activities of crystalline pertussigen from Bordetella pertussis. AB - We studied various biological activities of crystalline pertussigen and found that in mice as little as 0.5 ng of pertussigen induced hypersensitivity to histamine, 8 to 40 ng induced leukocytosis, 2 ng increased production of insulin, 0.1 ng increased production of immunoglobulin E and immunoglobulin G1 antibodies to hen egg albumin, 9.5 ng increased susceptibility to anaphylactic shock, and 0.5 ng increased the vascular permeability of striated muscle. We also found that in Lewis rats 20 ng of pertussigen promoted the induction of hyperacute experimental allergic encephalomyelitis. Pertussigen given intraperitoneally was toxic to mice at a dose of 546 ng. Treatment of pertussigen with glutaraldehyde eliminated this toxicity. Mice immunized with 1,700 ng of detoxified pertussigen were protected against intracerebral challenge with 3 x 10(4) viable Bordetella pertussis cells. When as little as 0.5 ng of pertussigen was given intravenously to mice, the increased susceptibility of the animals to histamine could still be detected 84 days later. The biological properties of crystalline pertussigen indicate its similarity to leukocytosis-promoting factor, Islet-activating protein, late-appearing toxic factor, and mouse-protective antigen of B. pertussis. PMID- 6270000 TI - Effects of cell source, mouse strain, and immunosuppressive treatment on production of virulent and attenuated murine cytomegalovirus. AB - Murine cytomegalovirus pools from various in vitro and in vivo sources were compared for virulence in suckling mice in an effort to identify the conditions which were necessary for the production of virulent and attenuated viruses. Virus passaged in tracheal ring and salivary gland organ cultures, where virus is produced primarily by epithelial cells, was even more attenuated than virus passaged in mouse embryo fibroblasts. The attenuation observed after passage in all three of these in vitro systems did not appear to be due to defective interfering particles. We also found that virus produced in vivo in salivary glands became attenuated with time after infection. Virus harvested from salivary glands 5 to 6 weeks after infection was highly attenuated compared with both salivary gland-passaged virus harvested 2 to 3 weeks after infection and tissue culture-passaged virus. The attenuation of salivary gland-passaged virus with time was reversed when animals were treated with cyclophosphamide before the virus was harvested. A comparison of virus pools harvested from susceptible and resistant mouse strains indicated that the mouse strain had little effect on the virulence of the virus produced. When the various sources of virus tested in this study were ranked in terms of the virulence of the virus produced, salivary glands in intact mice either 2 to 3 weeks after infection or after cyclophosphamide treatment produced the most virulent virus, followed by mouse embryo fibroblast cultures, tracheal ring and salivary gland organ cultures, and, finally, salivary glands in intact mice 5 to 6 weeks after infection. PMID- 6270001 TI - Factors affecting the rate as which a trachoma strain of Chlamydia trachomatis establishes persistent infections in mouse fibroblasts (McCoy cells). AB - Chlamydiae from mouse fibroblasts (McCoy cells) persistently infected with a trachoma strain (G17) of Chlamydia trachomatis repeatedly established new persistent infections in wild McCoy cells more rapidly than did wild chlamydiae. The density and nutritional state of the host cell populations may also influence the rate of establishment of persistent infections. PMID- 6270002 TI - Distinctive ribonucleic acid patterns of human rotavirus subgroups 1 and 2. AB - The ribonucleic acid migration patterns of 7 subgroup 1 and 16 subgroup 2 human rotaviruses recovered from four geographic areas were compared. The subgroup 1 ribonucleic acid patterns had strikingly slower-moving segments 10 and 11, suggesting a correlation between the ribonucleic acid pattern and the subgroup specificity. PMID- 6270003 TI - Oxygen toxicity: etiology. PMID- 6270004 TI - Identification of luteinizing hormone-releasing factor and alpha subunit of glycoprotein hormones in ductal carcinoma of the mammary gland. AB - The immunoperoxidase technique was used to identify luteinizing hormone-releasing factor (LRF) and the alpha subunit of glycoprotein hormones in ductal carcinoma of the mammary gland. Eleven tumors were studied, using antisera against LRF and alpha subunit, and against the beta subunits of hCG, LH, FSH and TSH. Six of the tumors were both LRF- and alpha-positive and three were negative. One tumour was LRF-positive/alpha-negative, and one was LRF-negative/alpha-positive. None of the anti-beta subunit antisera gave positive staining, and normal breast tissue as well as chronic cystic mastopathy (five cases) were negative with all antisera tested. The results with the anti-alpha and anti-beta subunit antisera suggest that the material stained is free alpha subunit rather than any of the native glycoprotein hormones which share the common alpha subunit. PMID- 6270005 TI - Re-evaluation of a transforming strain of Epstein-Barr virus from the Burkitt lymphoma cell line, Jijoye. AB - The biological properties of Epstein-Barr virus (EBV) from a Burkitt lymphoma cell line, Jijoye, were examined. the synthesis of virus capsid antigen (VCA) and early antigen (EA) in Jijoye cells was markedly enhanced by shift-down of the temperature of incubation from 37 degrees C to 33 degrees C. Cultures of Jijoye cells at 33 degrees C released a large amount of transforming EBV (10(5.2) of 50% transforming doses/ml) into the culture fluid. However, no infectious virus was produced in all cultures at 37 degrees C during the course of this study. The EBV (Jijoye EBV) from Jijoye line was found to possess only transforming activity, but not EA-inducing activity. Jijoye EBV lacks adsorbing capacity to Jijoye cells, in contrast to P3HR-I EBV which can adsorb to Jijoye cells. The Jijoye cells were highly susceptible to superinfection with P3HR-I EBV as demonstrated by the induction of EA, VCA and infectious EBV. The EBV induced by the P3HR-I EBV superinfection of Jijoye cells has also transforming activity but neither EA inducing activity nor adsorbing capacity to Jijoye cells. PMID- 6270006 TI - Multiplicity-dependent biological and biochemical properties of Epstein-Barr virus (EBV) rescued from non-producer lines after superinfection with P3HR-1 EBV. AB - Superinfection of lymphoblastoid cells of EBV non-producer lines with non transforming P3HR-1 EBV leads to the rescue of transforming virus. At least part of the recovered molecules represent recombinant DNA between superinfecting P3HR 1 EBV and resident viral genomes (Fresen et al., 1979, 1980). With high titer stocks of superinfecting P3HR-1 EBV, viral particles with early antigen (EA) inducing properties can be rescued, indicating that under these conditions of infection input viral genomes may become replicated. Sequential blot analysis with 32P-P3HR1-EBV DNA of intracellular viral DNA synthesized following infection reveals a multiplicity-dependent pattern. High particle inputs lead to preferential synthesis of certain fragments, independent of infection of either EBV genome-free or EBV genome carrier cells. This accumulation of specific viral sequences indicates defective replication of P3HR-1 EBV DNA. PMID- 6270007 TI - Induction of the EBV cycle in B-lymphocyte-derived lines is accompanied by increased natural killer (NK) sensitivity and the expression of EBV-related antigen(s) detected by the ADCC reaction. AB - Human B-lymphocyte-derived lines were forced to enter the EBV-cycle by superinfection with the P3HR-1 substrain of EBV or sodium butyrate treatment. The induced cells were used as targets for natural killing (NK) and EBV-specific, antibody-dependent cellular cytotoxicity (ADCC). Two Burkitt lymphoma lines, Raji and Daudi, and one normal adult derived lymphoblastoid cell line, NAD-7, were comparable in their ADCC-sensitivity after induction, but only the Burkitt lymphoma-derived lines showed a major increase in NK-sensitivity. The superinfection-induced membrane change, responsible for both NK and ADCC sensitivity, is an early function of the viral cycle, correlated with the appearance of early antigens (EA). Indirect evidence indicates that the NK and ADCC target sites are different but this problem requires further investigation. Sodium butyrate induced an increased NK sensitivity and EBV-related ADCC sensitivity in the Burkitt lymphoma-derived P3HR-1 line. Lymphocyte effectors from different donors showed great differences in their NK and ADCC activity. Optimal ADCC could be demonstrated with effectors that were intermediate in their NK-activity. PMID- 6270008 TI - Evidence of separate pathways for viral and chemical carcinogenesis in C3H/StWi mouse mammary glands. PMID- 6270009 TI - Detection of natural killer cells in Japanese quails. AB - Spleen cells of normal quails exhibited cytotoxicity on normal embryo cells as well as on virus- and chemically transformed cells. Spleen cells responsible for the destruction of target cells were shown to be non-adherent lymphoid cells which lack surface immunoglobulin and are resistant to the treatment with anti thymocyte serum and complement. Spleen cells of quails which were injected with chicken amniotic fluid (ChAmF) showed no cytotoxic activity. Furthermore, the addition of ChAmF-treated spleen cells abolished the cytotoxic activity of untreated spleen cells. These results indicated the presence of a spleen-cell population in normal quails compatible with mammalian natural killer (NK) cells and the induction of suppressor cells directed to the NK cells by administration of ChAmF. PMID- 6270010 TI - Analysis of normal neoplastic human tissues for the tumor-associated protein p97. AB - We have used immunoprecipitation to test tumor biopsies and normal adult and fetal human tissues for p97, a tumor-associated protein. Five of nine melanoma biopsies contained p97 in low to very high levels. Three of seven non-melanoma tumors contained p97, but in smaller amounts. No p97 was detected in any of the normal adult tissues examined. The protein was, however, observed in samples of fetal colon and umbilical cord, and in one sample of fetal lung. One of two benign nevi contained high levels of p97, whole on benign angiofibroma was negative. We conclude that the presence of p97, in levels detectable by our method, appears to be characteristic of certain neoplastic and fetal tissues. PMID- 6270011 TI - Enhancement of outgrowth of EB virus-transformed cells from normal human peripheral blood by a tumor promoter, TPA. AB - Effect of 12-0-tetradecanoyl-phorbol-13-acetate (TPA) on the establishment of lymphoblastoid cell lines (LCL) following spontaneous tranformation of B cells by Epstein-Barr virus (EBV) from peripheral lymphocytes of healthy EBV-seropositive adults was examined. In the lymphocyte culture with TPA (0.5 ng/ml), the frequency of establishment of LCL increased and the time required for appearance of transformation decreased. A larger number of EBV-positive cells in the peripheral blood were detected in the assay by co-cultures with cord blood lymphocytes in the presence of TPA than in the absence of it. The enhancement of LCL establishment by TPA was largely abolished in the culture with EBV neutralizing antibody-containing human umbilical cord serum. This suggests that the enhancement by TPA may be caused mostly by transformation of coresident B lymphocytes infected with active virus induced from the EBV genome-positive cells by TPA. PMID- 6270012 TI - EBV infection of mitogen-stimulated human B lymphocytes. AB - B lymphocytes from human blood were treated with the mitogen Protein A (Staph, aureus), which induces DNA synthesis in, as well as differeniation of, the B lymphocytes. The cells were subsequently exposed to Epstein-Barr virus (EBV). EBV infected and mitogen-stimulated cells were detected simultaneously, by using a combination of immunofluorescence (for EBNA) and autoradiography (for DNA synthesis). In the initial phase (1-2 days) after addition of the mitogen, stimulated cells were as susceptible to infection as resting cells. Thereafter, their susceptibility decreased. This suggests that, although initiation of DNA synthesis does not seem to limit sensitivity to the viral infection, differentiation of the B cells might do so. The intensity and pattern of EBNA staining in prestimulated cells different from that of the controls. PMID- 6270013 TI - Naturally occurring humoral immunity to endogenous xenotropic and amphotropic type-C virus in the mouse. AB - Natural humoral cytotoxic antibodies from 13- and 18-month-old BALB/c mice showed a virus-specific complement-dependent activity against target cells productively infected with xenotropic, amphotropic or ecotropic type-C viruses. The cytotoxic activity was lowest against ecotropic virus-shedding cells. Serum obtained from mice less than 12 months old had no such reactivity. The cytotoxic reactivity was found to reside solely in the immunoglobulin M fraction which yielded reactivity comparable to the unfractionated sera concerning both titer and relative reactivities to the target cells infected with different type-C viruses. In hyperimmune mouse serum cytotoxic reactivity resided in IgG and IgM fractions. Examination of serum from individual, normal 18-month-old BALB/c mice revealed that 80-90% of them were cytotoxic against virus non-producer mink target cells expressing gp70 or gaggene product. Absorption of sera from 18-month-old normal BALB/c mice with cells shedding Class II or Class III xenotropic virus, amphotropic virus, Rauscher-MuLV, or Class 1 murine leukemia virus indicate a closer amphotropic-FMR viral subtype specificity of the natural cytotoxic immune response as compared to the amphotropic-xenotropic or amphotropic-ecotropic specificity. The incidence and the level of measured humoral cytotoxic activity was sustained in the tumor-bearing animal up to 28 months of age as compared to the background established in the 18-month-old animal. However, in the non-tumor bearing animal, the incidence and level of cytotoxic reactivity declined rather rapidly with aging. Te sustained cytotoxic reactivity of the serum from old tumored mice might be involved in the progression of the tumor. PMID- 6270014 TI - Difference in viral binding between two Epstein-Barr virus substrains to a spectrum of receptor-positive target cells. AB - Radio-labelled Epstein-Barr virus (EBV) was utilized in a direct binding assay to detect the presence of EBV receptors. The sensitivity of this method was affirmed by the detection of EBV-receptors on three EV-carrying cell lines that have previously been reported as receptor negative. Two laboratory substrains of EBV, derived from the cell lines B95-8 and P3HR-I (designated B and P virus respectively), were tested in the binding assay. The main repcptor prototype adsorbed both viral strains without apparent distinction. In contrast, two lines, a Swedish EBV-negative B-cell lymphoma (U698) and a virus non-producer subline of the receptor-negative P3HR-1 line, adsorbed P virus selectively but failed to adsorb B virus. PMID- 6270015 TI - Demonstration of the transferrin receptor in human breast cancer tissue. Potential marker for identifying dividing cells. AB - A transferrin receptor was demonstrated in tumor tissue from 10 patients with breast carcinoma and one patient with breast sarcoma. Binding studies were conducted by measuring the amont of 1251-transferrin binding to microsomal preparations of the tumor tissue. Elevated levels of specific transferrin binding were found in the tumors with a range of 11-35% of bound transferrin, whereas microsomes prepared from non-neoplastic breast tissue samples bound only 2.3% and 2.4% of the transferrin. Scatchard analysis of binding studies conducted with tissues from a breast cancer and from a breast sarcoma indicate that the receptor has a Ka = 9.0 x 10(8)M. The binding site is specific for transferrin, as studies show that non-radioactive transferrin displaced labelled transferrin, while human IgG and human albumin did not. The receptor-transferrin complex was precipitated from a detergent extract of the breast sarcoma with antiserum to human transferrin. Sodium dodecyl sulfate/polyacrylamide gel electrophoresis of the immunoprecipitate gave a polypeptide of M 90,000 daltons, which is of similar molecular weight found for the putative transferrin receptor in all of a series of human cultured cell lines previously examined. PMID- 6270016 TI - Host range and oncogenicity studies of the murine myeloma MOPC-315 type-C retrovirus. AB - The biological features of an endogenous type-C RNA tumor virus released by the murine plasmacytoma MOPC-315 were determined. The virus was found to be pure ecotropic and, according to its capability to infect both NIH/3T3 and BALB/3T3 cells, it was established as an NB tropic virus. This tropism became more pronounced as a consequence of multiple passage in balb/3T3 cells. In view of the ability of myeloma viruses to efficiently infect BALB/c cells, their possible role in myeloma induction is considered. PMID- 6270017 TI - In vivo formation of benzo(alpha)pyrene diol epoxide-deoxyadenosine adducts in the skin of mice susceptible to benzo(alpha)pyrene-induced carcinogenesis. AB - The hydrocarbon-deoxyribonucleoside adducts formed in mouse skin DNA have been determined following topical application of an initiating dose of benzo(a)pyrene to Swiss mice, a strain shown to be susceptible to benzo(a)pyrene-induced skin carcinogenesis. Several DNA-bound products were formed, of which the major one (60% of total adducts), in agreement with other workers' findings, was derived from reaction of (+/-) 7 beta, 8 alpha-dihydroxy-9 alpha, 10 alpha-epoxy-7,8,9,10 tetrahydrobenzo(alpha)pyrene (BDE) with the exocyclic aminogroup of deoxyguanosine. A further product (9-10% of total adducts), previously observed only after microsomal activation of benzo(a)pyrene, was observed and co chromatographed with a further metabolite of 9-hydroxybenzo(alpha)pyrene bound to an uncharacterized base in the DNA. Two otherr products (2-3% of total adducts) were also found in the in vivo studies which co-chromatographed with BPDE deoxyadenosine adducts and arose from cis and trans addition of the exocyclic amino group of deoxyadenosine to the 7R form, but not the 7S form, of BPDE. In contrast to this, the major in vitro deoxyadenosine-bound products, formed following reaction of BPDE with calf-thymus DNA, were derived from the 7S form of BPDE, suggesting either stereoselective formation or reaction of the 7R form of BPDE in mouse skin in vivo. Similar amounts of BPDE-deoxyguanosine and BPDE deoxyadenosine adducts, as well as those derived from further metabolism of 9 hydroxybenzo(alpha)pyrene were formed in three strains of mice reported to have widely differing susceptibilities to polycyclic hydrocarbon-induced skin carcinogenesis. The relevance of these different hydrocarbon-DNA adducts to carcinogenesis requires further investigation. PMID- 6270018 TI - Genetic resistance to herpesvirus-induced lymphoma at the level of the target cell determined by the thymic microenvironment. PMID- 6270019 TI - Mechanisms of enhancement of Epstein-Barr virus-induced transformation of peripheral blood lymphocytes by a tumor promoter, TPA, with special reference to lowering of cytotoxicity of T cells. AB - The mechanism of the previously observed enhancement of Epstein-Barr virus (EBV) induced cell transformation of human lymphocytes by 12-0-tetradecanoyl-phorbol-13 acetate (TPA) was studied. A concentration of TPA (0.5 ng/ml) was used. In cultures of human cord blood lymphocytes infected with EBV in the presence of TPA, a larger number of EBV-associated nuclear antigen (EBNA)-positive and/or DNA synthesizing cells was observed than in the absence of TPA. In the virus-infected lymphocyte cultures of EBV-seropositive adult donors, in vitro regression of transformation, which is known to be caused by T lymphocytes, was suppressed by TPA, EBV-specific and -non-specific cytotoxicity of T cells generated in mixed cultures of peripheral blood lymphocytes (PBL) of seropositive adults and autologous lymphoblastoid cell line (LCL) cells was markedly lowered by the presence of TPA in the cultures, However, TPA had little effect on proliferation of T cells in stimulated cultures, and addition of TPA to the reaction mixture for the cytotoxicity test did not lower the cytotoxicity. These results suggest that TPA has an inhibitory effect on T cells which suppress EBV-transformation. THe effect on T cells, together with the direct promoting effect on proliferation of EBV-transformed cells, may explain the outcome of enhancement of EBV-induced LCL establishment from PBL of seropositive donors by TPA previously observed. PMID- 6270020 TI - Characterization of a human papillomavirus from epidermodysplasia verruciformis lesions of a patient from Upper-volta. AB - A case of epidermodysplasia verruciformis in a patient from Upper-Volta is described. Slightly elevated, flat warts were observed on hands, feet, arms and legs, and pityriasis versicolor-like lesions were found mainly on the trunk. The patient showed no malignant tumors. Histological examination revealed hyperkeratosis, granulosis and moderate acanthosis with large, foamy, basophilic keratinocytes in stratum granulosum and stratum spinosum. Papillomavirus particles could be prepared from these lesions and were differentiated from known papillomavirus types by immune electron microscopy with monospecific antisera and by DNA-DNA hybridization. The viral DNA was characterized by cleavage with several restriction endonucleases and a physical map of the resulting fragments was established. The virus is designated as HPV 8. Preliminary seroepidemiologic studies with human sera indicate a rather wide distribution of HPV 8. Blot hybridization of DNA from human carcinomas with 32P-labelled virus DNA detected no HPV 8-specific sequences. PMID- 6270021 TI - Cytomegalovirus: detection in human colonic and circulating mononuclear cells in association with gastrointestinal disease. AB - The specificity and strength of the reported association between cytomegalovirus (CMV) and colonic adenocarcinoma were tested by analysis of a consecutive series of surgically resected colons for: CMV-DNA by a DNA-DNA reassociation kinetics (hybridization) procedure; latent virus by co-cultivation of fresh tissue with indicator fibroblasts; and CMV viral antigens by immunofluorescence. Ten of 13 cancer patients whose colonic tissue was able to be examined by all techniques showed evidence of active or prior CMV infection. Four cancer specimens were CMV DNA (hybridization)- positive; an additional specimen from a cancerous colon was culture-positive. In six instances, CMV DNA was detected in mucosal cells adjacent to colon adenocarcinoma. In tissue from one of three patients with ulcera tive colitis and two of seven patients with other non-neoplastic colonic disease, CMV DNA was also detected. No fresh colonic tissues were demonstrated to have CMV surface or nuclear antigens when examined by immunofluorescence. Culture of peripheral lymphocytes was positive for CMV in three of 14 cancer patients. A CMV specific defect in humoral immunity could not be documented in that most cancer patients, as well as cancer-free patients, exhibited circulating specific antibody to CMV and had a normal capacity for CMV-specific antibody-dependent cellular cytotoxicity. We conclude that CMV, probably in a latent form, is readily detectable in colonic cells of man, including those derived from malignant, pre-malignant and non-malignant tissues. Neither preferential replication in damaged tissue nor carriage of CMV by peripheral lymphocytes homing to gut appear to explain the presence of CMV in colon cells. PMID- 6270022 TI - Class-specific antibodies (IgG and IgA) to membrane antigens of Herpes simplex type 2-infected cells in patients with cervical dysplasia and neoplasia. PMID- 6270023 TI - Detection and characterization of viral genomes and search for tumoral antigens in two hamster cell lines derived from tumors induced by bovine papillomavirus type 1. PMID- 6270024 TI - Extracellular matrix proteins characterize human tumor cell lines. AB - Extracellular matrix proteins synthesized and secreted by adherent human tumor cell lines were analyzed using metabolic labelling with glycine and proline in the presence of ascorbate, polypeptide analysis and polyacrylamide gel electrophoresis, affinity chromatography, collagenase digestion, and immunofluorescence staining. The results showed a characteristic pattern of matrix proteins for each tumor cell type. Tumor cell lines of mesenchymal origin produced mostly interstitial types (I and II) of collagen and fibronectin. Carcinoma cell lines secreted only basement membrane proteins, type IV collagen, laminin and fibronectin, but not interstitial collagen. A melanoma and a rhabdomyosarcoma cell line produced type V of procollagen that has not previously been described in cell culture. Neuroblastoma cells were shown to be phenotypically heterogeneous also with respect to matrix protein production. We propose that the analysis of extracellular matrix proteins may serve as an adjunct in the classification of human tumors. PMID- 6270025 TI - Lack of induction of murine mammary tumor virus expression in cultured mammary glands treated with chemical carcinogens. PMID- 6270026 TI - Helper virus-dependent RSV rescue from two lines of hamster cells transformed in vivo by XC RSV. AB - Newborn hamsters were inoculated with chicken sarcoma tissue produced by XC RSV which had been rescued by transfection from XC cells. Tumours with a latency of up to 20 months appeared in all inoculated hamsters. Three tumour cell lines were successfully established for in vitro growth and cloned. The virus was rescued from the PR RSH-9 cell line by cell fusion with chicken fibroblasts and from the other two lines (PR RSH-12 and PR RSH-14) and clones only if chicken fibroblasts used for fusion had been preinfected with a helper virus. The last two cell lines evidently harboured a functional env gene since the the synthesis of transforming virus started soon after they were found with 16Q cells producing defective BH RSH(-) lacking the envelope glycoprotein. Produced viruses acquired subgroup C specificity of XC rsv originally used for transformation of these two cell lines. The nature of provirus alterations in PR RSH-12 and PR RSH-14 cells as well as the mechanisms underlying these changes are discussed. PMID- 6270027 TI - The effect of infection on T lymphocyte subpopulations: a preliminary report. AB - The effect of cytomegalovirus (CMV) infection on T-lymphocyte subpopulations (employing the monoclonal antibody OKT4 to delineate helper cells and OKT8 for suppressor-cytotoxic cells) was studied in 10 normal individuals with CMV mononucleosis and four renal transplant patients with symptomatic primary CMV disease. In both clinical settings, acute CMV infection is associated with a reversal of the normal helper/suppressor-cytotoxic T cell ratio, with both a relative and absolute decrease in T helper cells and an increase in T suppressor cytotoxic cells observed. With clinical recovery, there is a return of these cell populations toward normal ranges. In two patients, one with chronic CMV infection and one with chronic Epstein-Barr virus infection, the helper/suppressor cytotoxic cell ratio was found to be reduced four years after the onset of infection. Five patients with febrile illnesses were also studied: two with viral infection and one with a self-limited febrile illness had decreased helper/suppressor-cytotoxic cell ratios, and two with bacterial infection had increased ratios. Thus, delineation of T lymphocyte subpopulations may be of value not only in evaluating the immunologic effects of a particular infectious agent, but also in the clinical evaluation of the febrile patient. PMID- 6270028 TI - Papillomavirus and cutaneous malignancy. PMID- 6270029 TI - Polio and lameness in Yogyakarta, Indonesia. AB - This survey for residual paralysis among rural and urban children in Central Java shows an overall rate for polio lameness of 9/10 000 children. A recent rise in reported cases of acute poliomyelitis is reflected in paralysis rate among preschool children of 22/10 000. Three quarters of cases occur before the third birthday. A majority of children below age 3 are lacking antibody titres to one or more types of polio virus, demonstrating the need for mass immunisation. A simple questionnaire detected polio cases with a sensitivity of 84%. PMID- 6270030 TI - Hydrolysis of soybean trypsin inhibitor by the gamma subunit of 7SNGF and EGF-BP. AB - Although soybean trypsin inhibitor (STI) does not inhibit the esterase activity of either epidermal growth factor binding protein (EGF BP) or the gamma subunit of 7SNGF, it does behave as a substrate for proteolysis. Cleavage of the active site peptide bond of STI does occur when incubated in the presence of either EGF BP or the gamma subunit of 7SNGF. The hydrolysis id pH dependent with maximum proteolysis at pH 6.0-7.0. the newly formed C-terminal arginine residue in modified STI can be released by carboxypeptidase B digestion. Both enzymes are inhibited by low concentrations (2-4 microgram/ml) of the microbial protease inhibitors leupeptin and antipain. These inhibitors are specific for trypsin-like proteases. Since both enzymes can be found as part of high molecular weight complexes with growth factors these results confirm the hypothesis that they are involved during a postranslational modification event. PMID- 6270031 TI - Free radicals from X-irradiated single crystals of uridine-5'-phosphate disodium salt. AB - X-irradiation of single crystals of uridine-5'-phosphate (disodium salt) between 10 and 300 K as well as storage of irradiated crystals at 300 K produces at least seven different radical species. Between 10 and 77 K, the uracil base anion and a secondary alkoxy radical at the ribose-O3'-site are formed. The latter transforms into a C5'-centred alkylphosphate species between 110 and 130 K which in turn decays between 180 and 220 K under formation of a base 5-yl hydrogen addition radical. Irradiation at 300 K additionally produces the base-located 6-yl radical together with a radical tentatively assigned to the doubly protonated base anion. Storage of crystals for several months results in decay of most of these species leaving a radical possibly located at c5' of the ribose. The spectral parameters of these radicals are given and discussed. PMID- 6270032 TI - Globin-mediated redox processes in the interaction of hydroxyl radicals with ferrimyoglobin in aqueous solution. PMID- 6270033 TI - The influence of alkali metal chlorides on environmentally-linked behavioural stereotypies in the rat. AB - In a series of three experiments using rats, the chlorides of lithium, rubidium and caesium were compared with sodium chloride in respect of their effects on behavioural stereotypy. Two apparatuses were used, a modified hold-board and an open field. Caesium was found to increase the occurrence of environmentally linked behaviour in both types of apparatus. Lithium had effects in the opposite direction. Rubidium had effects like caesium in the modified hold-board test, but like lithium in the open field. The findings are discussed in terms of possible effects on central information processing mechanisms. PMID- 6270034 TI - The effects of lithium on the rate of recovery of rats from ethanol-induced narcosis. PMID- 6270035 TI - Cryopreservation of varicella-zoster virions without loss of structural integrity or infectivity. AB - Varicella-zosterer virions present in infected cells or in a cell-free state were freeze-dried without loss of structural integrity of infectivity. Generally, yields of greater than 5 log10 foci/ml (infected cells) or greater than 4 log10 PFU/ml (cell-free virus) were recovered from varicella-zoster virus-infected human melanoma cells both before and after lyophilization in phosphate-buffered media containing 0.1-1.0 M sucrose. Virus frozen in solutions lacking sugar had little or no residual infectivity after vacuum sublimation was completed. Visualization by electron microscopy demonstrated large numbers of enveloped virions in the virus preparations lyophilized in media containing sucrose; in marked contrast, virus subjected to freeze-drying in buffered solutions without sugar consisted mainly of naked nucleocapsids. Water analyses by Karl Fischer titration suggested that residual moisture retained by sugar prevented disenvelopment of the varicella-zost virion. PMID- 6270036 TI - Pseudohypoparathyroidism with normal phosphaturic response to exogenous parathyroid hormone administration. PMID- 6270037 TI - A survey of virus particles in infantile gastroenteritis using the electron microscope. PMID- 6270038 TI - 1980 Beaumont Lecture. Restriction enzymes as tools for genetic research. PMID- 6270039 TI - Cushing's disease, a pituitary disorder. A hypothesis. PMID- 6270040 TI - Louisa Burns Memorial Lecture 1980: The spinal cord -- active processor not passive transmitter. PMID- 6270041 TI - [Care of parents with prenatal fetal death]. PMID- 6270042 TI - The critical incident technique applied to the evaluation of the clinical practicum setting. PMID- 6270043 TI - Clinical instruction in nursing: a national survey. PMID- 6270044 TI - Self-actualization and professional socialization of nursing students in the clinical laboratory experience. PMID- 6270045 TI - Experience in posology. AB - We used an organized problem-solving method to resolve posology problems in integrated curriculum. A thorough assessment of the extent of problems revealed the need for a mathematical foundation (perhaps due to decreased mathematics SAT scores) to achieve a high level of accuracy with posology in preparation for professional nursing practice. Early curriculum intervention allowed for higher learning level achievement and sufficient time for demonstration of acquired skills. The implementation of a self-learning module in conjunction with small group intervention appeared to have an impact on students' level of knowledge as well as the application of knowledge. This was evidenced by an increased mean score from pre- to post-testing on a multiple-choice examination. Ongoing evaluations will measure students' ability to actualize these concepts in their clinical practice. Perhaps now is the time to identify posology within integrated curricula and examine it as a basis for pharmacology to better prepare all students for this vital nursing responsibility. PMID- 6270046 TI - Acute idiopathic polyneuropathy in the dog. AB - From among a large group of dogs with acute tetraparesis, we identified 10 dogs with a distinct peripheral nerve disorder. Prior to the onset of signs, all of the dogs had been healthy, and none was known to have been exposed to a neurotoxin or raccoon bite. Weakness, with hypoactive or absent segmental reflexes, became progressively worse for 1 to 21 days. Results of electromyography and nerve conduction studies invariably were compatible with a diagnosis of polyneuropathy that predominantly affected proximal nerve segments. Appearance of nerve biopsy specimens and the short time course for functional recovery suggested a demyelinative component to the disorder. The extent of recovery was variable but often rapid and complete in dogs that did not succumb to complications in the early period. Corticosteroid therapy did not demonstrably influence the outcome. This acute idiopathic polyneuropathy in the dog shares many clinical and pathologic features with idiopathic polyradiculoneuritis (Coonhound paralysis). PMID- 6270047 TI - Enzymatic adenylylation of spectinomycin by Acinetobacter calcoaceticus subsp. anitratus. AB - Spectinomycin (SPC) was inactivated in the presence of adenosine-5'-triphosphate and magnesium ion by an enzyme preparation made from Acinetobacter calcoaceticus subsp. anitratus GN12313 resistant to both SPC and streptomycin. The structure of the inactivated SPC was found to be the adenylylated product of the hydroxy group on C-9 of the actinamine moiety. PMID- 6270048 TI - The structures of component A1 (= LL-AB664) and component A2 (= LL-AC541), streptothricin-like antibiotics. PMID- 6270049 TI - Antipseudomonal beta-lactams. PMID- 6270050 TI - Occurrence of aminoglycoside-modifying enzymes in resistant strains of enterobacteria and Pseudomonas aeruginosa from several countries. PMID- 6270051 TI - Comparative in-vitro activity and mode of action of ceftriaxone (Ro 13-9904), a new highly potent cephalosporin. PMID- 6270052 TI - Susceptibility of Pseudomonas aeruginosa to cefoperazone, cefotaxime and moxalactam, with special reference to isolates resistant to aminoglycosides, carbenicillin and ticarcillin. PMID- 6270053 TI - In-vitro efficacy of cefotiam in combination with four aminoglycosides on staphylococci and Gram-negative bacteria. PMID- 6270054 TI - Temperature-programmed gas chromatographic determination of polychlorinated and polybrominated biphenyls in serum. AB - An analytical method was developed to quantitate polychlorinated and polybrominated biphenyls (PCBs and PBBs, respectively) in human serum. The method includes denaturation of the proteins in serum, extraction, adsorption chromatography, and gas chromatography with electron capture detection. The coefficients of variation for determining the in vivo bound PCBs and PBBs ranged from 11.7 to 29.8% and 7.1 to 14.0%, respectively. The method is capable of measuring 10 ng PCBs and PBBs/mL in 4 mL serum. PMID- 6270055 TI - Early initiation of deoxyribonucleic acid replication and shortening of generation time associated with inhibition of lateral wall formation by mecillinam. AB - The effects of mecillinam on the growth of rods of the pH-conditional morphology mutant MirM7 was studied. It has been found that mecillinam causes, coincident with transition to coccal shape, a balanced rise in the rate of viable count increase and the rate of macromolecular synthesis which lasts either until the cells enter a stationary growth phase or indefinitely, in the case of continuously diluted cultures. When the antibiotic is removed from cells which have already become coccoid, cells continue to grow at a faster rate until they resume the rod shape. No change in the per-cell rate of protein synthesis has been seen in untreated or mecillinam-treated cells before or after the change in growth rate. Studies with synchronously growing cells have shown that the antibiotic causes a shortening in the I period (initiation of deoxyribonucleic acid replication). Evaluation of the residual divisions in nalidixic acid treated, exponential-phase cells has shown that mecillinam also shortens the D period (cell division). It is proposed that, in strain MirM7, inhibition of lateral wall elongation by the antibiotic allows the initiation of a new septum, though inhibition is still in progress. The initiation of a new septum is, in turn, responsible for both the early inibition of deoxyribonucleic acid replication and accelerated division. In the parental strain, MirA12, as well as in other sensitive gram-negative rods which divide, become cocci, and stop dividing after addition of the antibiotic, inhibition of lateral wall formation activates a feedback mechanism which prevents insertion of new septa (Satta et al., J. Bacteriol. 142:43-51, 1980). Consequently, no early initiation of deoxyribonucleic acid replication is observed, and the last division allowed by the antibiotic occurs in due time. This negative control is missing in MirM7. PMID- 6270056 TI - Integration of the bacteriophage phi 3T-coded thymidylate synthetase gene into the Bacillus subtilis chromosome. AB - Transformation of Bacillus subtilis 168 Thy- auxotrophs with phi 3T deoxyribonucleic acid (DNA) to thymine independence was found to involve site specific recombination of phi 3T DNA sequences with their homologous counterparts in the bacterial chromosome. During the transformation, the phage phi 3T-encoded thymidylate synthetase gene, thyP3, was shown to integrate at two genetically distinct sites in the B. Subtilis 168 chromosome. The first site was identified to be in the bacterial thymidylate synthetase gene, thyA. The second site was in a prophage (SPB) known to be carried in the host genome. The frequency of the integration of the thyP3 gene at each of the two loci and some of the parameters affecting this frequency were studied. The common origin of the thyP3 and thyA genes and their molecular evolution are also reported. PMID- 6270057 TI - Adenosine 5'-triphosphate- yielding pathways of branched-chain amino acid fermentation by a marine spirochete. AB - The metabolic pathways utilized by an obligately anaerobic marine spirochete (strain MA-2) to ferment branched-chain amino acids were studied. The spirochete catabolized l-leucine to isovaleric acid, l-isoleucine to 2-methylbutyric acid, and l-valine to isobutyric acid, with accompanying CO(2) production in each fermentation. Cell extracts of spirochete MA-2 converted l-leucine, l-isoleucine, and l-valine to 2-ketoisocaproic, 2-keto-3-methylvaleric, and 2-ketoisovaleric acids, respectively, through mediation of 2-ketoglutarate-dependent aminotransferase activities. The branched-chain keto acids were decarboxylated and oxidized to form isovaleryl coenzyme A, 2-methylbutyryl coenzyme A, and isobutyryl coenzyme A, respectively, in the presence of sulfhydryl coenzyme A and benzyl viologen. The acyl coenzyme A's were converted to acyl phosphates by phosphate branched-chain acyltransferase enzymatic activities. Branched-chain fatty acid kinase activities catalyzed formation of isovaleric, 2-methylbutyric, and isobutyric acids from isovaleryl phosphate, 2-methylbutyryl phosphate, and isobutyryl phosphate, respectively. Adenosine 5'-triphosphate was formed during conversion of branched-chain acyl phosphates to branched-chain fatty acids. The results indicate that conversion of l-leucine, l-isoleucine, and l-valine to branched-chain fatty acids by spirochete MA-2 results in adenosine 5' triphosphate generation. The metabolic pathways utilized for this conversion involve amino acid amino-transferase, 2-keto acid oxidoreductase, phosphate acyltransferase, and fatty acid kinase activities. PMID- 6270058 TI - Restriction map of corynebacteriophages beta c and beta vir and physical localization of the diphtheria tox operon. AB - The BamHI, EcoRI, HindIII, and KpnI restriction endonuclease maps of corynebacteriophage beta c and beta vir were constructed. beta vir appeared to be identical to beta c, except for an approximate 1-kilobase deletion that removed a BamHI site, two KpnI sites, and three EcoRI restriction sites. The diphtheria tox operon was located by hybridizing in vitro 32P-labeled tox messenger ribonucleic acid to blots of endonuclease-digested beta deoxyribonucleic acids. The messenger ribonucleic acid probe was found to hybridize to a 2.1-kilobase region of the beta genome. Since approximately 1.9 kilobases is required to encode prodiphtheria toxin, the data presented strongly suggest that the tox operon of beta is monocistronic. PMID- 6270059 TI - Physical mapping of beta-converting and gamma-nonconverting corynebacteriophage genomes. AB - Deoxyribonucleic acids (DNAs) from wild-type and mutant strains of beta converting and gamma-nonconverting corynebacteriophages were isolated and physically characterized. The data obtained from DNA heteroduplexes, restriction enzyme banding profiles, and restriction maps reinforce the conclusion that beta and gamma phages are very closely related. The major physical differences seen in the DNA heteroduplexes are a small substitution bubble and one or two insertions which are present on the gamma phage genome. The insertions account for the differences in the genome sizes of beta and gamma phages, and with the substitution they are responsible for most of the differences in the restriction endonuclease profiles and maps of the corynebactriophage genomes, two special sites and the DNA fragments carrying them were identified. These were the cohesive (cos) sites and the specific attachment (attP) site of the vegetative phage genome. The behavior of these sites indicated that the transition of phage DNA from the vegetative to the prophage state involves the circularization of vegetative DNA through the cos sites and its integration into the bacterial chromosome via the attP site. The mechanism of corynebacteriophage integration was similar to that employed by Escherichia coli phage gamma. From the data assembled the physical and genetic maps of beta and gamma phage were oriented with respect to one another. The extensive similarity in their maps provides additional confirmation of a close evolutionary relationship. PMID- 6270060 TI - Genetic elements novel for Corynebacterium diphtheriae: specialized transducing elements and transposons. AB - It was shown in an accompanying paper (Buck and Groman, J. Bacteriol. 148: 131 142, 1981) that gamma-tsr-1 phage stocks produced by heat induction of lysogens are a mixture of two phages which differ in the content of their deoxyribonucleic acid (DNA). This difference is evidenced by the appearance of "heterogeneous" (HET) fragments in restriction enzyme digests of gamma-tsr-1 phage DNA. It was estimated that 20 to 80% of the phage in these lysates produced HET fragments. The appearance of HET fragments correlated with the appearance of a DNA insertion (DI-1) in the gamma phage genome as revealed in heteroduplexes of DNA from gamma tsr-1 and beta corynebacteriophages. The HET fragments were seen in DNA from heat induced lysates, but not in DNA from phage stocks produced by lytic infection. By DNA-DNA hybridization analysis it was shown that a fraction of gamma-tsr-1 phages from heat-induced lysates carried an insertion of bacterial DNA in the vegetative phage attachment site (attP), and that this insertion was responsible for the formation of HET fragments. Since the phage produced by this event carried a complete phage genome plus a small segment of bacterial DNA, they were called transducing elements. On the basis of these facts it was concluded that heat induced gamma-tsr-1 prophage was excised at an abnormal site at a very high frequency. Abnormal excision was highly specific, and the change in excision specificity occurred simultaneously with the spontaneous mutation of the phage to heat inducibility. From this and other data it was postulated that a mutation in the immune repressor was reponsible for an alteration in the specificity of the normal excision process. This distinguishes the mechanism of formation of gamma tsr-1 transducing elements from that employed by other phages. A second DNA insertion (DI-2) in the tox (diphtheria toxin) gene of gamma-tsr-1 and gamma-tsr 2 was also identified as an insertion of bacterial DNA. The DI-2 insertion had a stem-and-loop structure similar to that seen in heteroduplexes visualizing transposons or insertion elements. It seems likely that gamma wild-type phage, which is mutant for tox, was originally tox(+), but that transposition of bacterial DNA into the gene inactivated it. PMID- 6270061 TI - Identification of deoxyribonucleic acid restriction fragments of beta-converting corynebacteriophages that carry the gene for diphtheria toxin. AB - Deoxyribonucleic acid fragments bearing the gene for diphtheria toxin have been identified in restriction enzyme digests of deoxyribonucleic acids from beta converting and gamma-nonconverting corynebacteriophages. A combination of physical and genetic evidence has established that the Bam HI band C fragment of beta phage deoxyribonucleic acid, which carries the specific phage attachment site (Buck and Groman, J. Bacteriol. 148:131-142, 1981), also carries most, and probably all, of the gene for diphtheria toxin. A detailed restriction map of this tox-bearing Bam HI fragment has been developed, and the locations and orientation of the tox gene and the attP site within this fragment have been established. PMID- 6270062 TI - Folic acid and pterin deaminases in Dictyostelium discoideum: kinetic properties and regulation by folic acid, pterin, and adenosine 3',5'-phosphate. AB - Kinetic data obtained for deamination of pterin by the extracellular fraction from Dictyostelium discoideum yielded apparently linear Lineweaver-Burk plots for pterin. The Michaelis constant for pterin was 30 microM. The data for folic acid deamination yielded convex Lineweaver-Burk plots. Convex Lineweaver-Burk plots could result from the presence of two types of enzymes with different affinities. The data for folic acid deamination were analyzed mathematically for two types of enzymes. This analysis produced Michaelis constants for folic acid of 1.8 and 23 microM competition studies suggested that an enzyme with low affinity nonspecifically catalyzed the deamination of folic acid and pterin, whereas an enzyme with high affinity was a specific folic acid deaminase. A specific folic acid deaminase with high affinity appeared to be present on the surface of D. discoideum cells. The Michaelis constant for this enzyme was 2.6 microM. Cells growing in nutrient broth and cells starved in phosphate buffer released folic acid and pterin deaminases. The quantity of deaminase activities released by the cells appeared to be controlled by chemoattractants. Starving cells that were supplied with folic acid, pterin, or adenosine 3',5'-phosphate increased their extracellular folic acid and pterin deaminase activities to a larger extent than did cell suspensions to which no chemoattractants were added. Administration of folic acid or pterin to starving cells caused increases of the activity of extracellular adenosine 3',5'-phosphate phosphodiesterase and repressed increases of the activity of phosphodiesterase inhibitor. PMID- 6270063 TI - Homology among tet determinants in conjugative elements of streptococci. AB - A mutation to tetracycline sensitivity in a resistant strain of Streptococcus pneumoniae was shown by several criteria to be due to a point mutation in the conjugative omega (cat-tet) element found in the chromosomes of strains derived from BM6001, a clinical strain resistant to tetracycline and chloramphenicol. Strains carrying the mutation were transformed back to tetracycline resistance with the high efficiency of a point marker by donor deoxyribonucleic acids from its ancestral strain and from nine other clinical isolates of pneumococcus and by deoxyribonucleic acids from group D Streptococcus faecalis and group B Streptococcus agalactiae strains that also carry conjugative tet elements in their chromosomes. It was not transformed to resistance by tet plasmid deoxyribonucleic acids from either gram-negative or gram-positive species, except for one that carried transposon Tn916, the conjugative tet element present in the chromosomes of some S. faecalis strains. The results showed that the tet determinants in conjugative elements of several streptococcal species share a high degree of deoxyribonucleic acid sequence homology and suggested that they differ from other tet genes. PMID- 6270064 TI - Mutagenesis by insertion of drug resistance transposon Tn7 into a vibrio species. AB - A halotolerant, collagenolytic strain of Vibrio sp. was conjugated with an Escherichia coli strain carrying plasmid RP4. The plasmid was transferred to and maintained in the Vibrio and could be subsequently transferred in matings to suitably marked stains of the same species. After conjugation with an E. coli carrying the cointegrate plasmid RP4::Mu cts61::Tn7, Vibrio transconjugants were selected that carried Tn7 inserted into the bacterial chromosome. A large proportion of these transconjugants were auxotrophic, showing that plasmid suicide by Mu can be used to isolate Tn7-derived mutants in Vibrio. Approximately half of the auxotrophs isolate Tn7-derived mutants in Vibrio. Approximately half of the auxotrophs isolated were ilv mutants, all of which exhibited the same phenotype. Thus, although Tn7 insertion can induce auxotrophy, including trp, thy, his and ura, in Vibrio, there does appear to be a hot spot for integration in the ilv operon. PMID- 6270065 TI - Variation in Escherichia coli buoyant density measured in Percoll gradients. AB - Escherichia coli B/r cells, centrifuged to equilibrium in either self-generating or preformed gradients of Percoll, banded at an average density of 1.080 to 1.100 g/ml, depending on their growth rate and the temperature of centrifugation. Cells cultured in gradient material (70% Percoll) exhibited the same average density. At the various growth rates examined, the density of the individual cells in a steady-state population varied by less than 1% of the mean in E. coli strains B/r and B, as well as K-12. Electron microscope analysis of the density fractions of both fast- and slow-growing E. coli B/r populations suggested a small increase in density during cell constriction. PMID- 6270066 TI - Protein phosphorylation and dephosphorylation in liver plasma membrane. Effect of inorganic phosphate. AB - When a plasma membrane preparation isolated from rat liver was incubated with [gamma-32P]ATP and Mg2+, protein-bound 32P increased rapidly, followed by a gradual decrease. The time course suggested the existence of membrane-bound kinase(s) and phosphatase(s) phosphorylating and dephosphorylating endogenous proteins. The extent of phosphorylation was not affected by inclusion of cyclic AMP in the reaction mixture. The extent of the maximum phosphorylation was dependent on membrane concentration, owing to rapid hydrolysis of ATP by the membrane-bound ATPase activity. Thus, phosphorylation proceeded further on repeated addition of ATP. Both phosphorylation and dephosphorylation were stimulated by Mg2+, an effective rate of phosphorylation being obtained at 15 mM. Pi up to 20 mM stimulated phosphorylation with little effect on the rate of dephosphorylation. At higher phosphate concentrations, the maximum 32P incorporation decreased again, and at 100 mM, dephosphorylation was prevented significantly. Autoradiography after polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate and urea revealed six main phosphorylated bands, two of which (Band 3 and 5) were partly extractable with 1 M NaCl. In the presence of 100 mM Pi, very strong phosphorylation of Band 5 (about 23,000 daltons) was noted, and a new strongly labeled band (Band P, about 20,000 daltons) was observed. It was concluded that the phosphoproteins in the membrane may be turned over at different rates and high concentrations of Pi may affect the turnover rate of some phosphoproteins, probably through interference with the phosphatase. PMID- 6270067 TI - Phosphoprotein phosphatase associated with rat liver plasma membrane. Properties of phosphorylase phosphatase and phosphohistone phosphatase. AB - Plasma membrane isolated from rat liver contained activities of phosphoprotein phosphatase dephosphorylating [32P]phosphorylase a or [32P]phosphohistone. The properties of the membrane-bound phosphatase were examined using these exogenous substrates. The optimal reaction rate was at pH near neutrality. At concentrations as low as 0.1-1.0 mM, Mg2+ or Mn2+ slightly stimulated the activity for phosphorylase a or phosphohistone, respectively; at higher concentrations, they were inhibitory with both substrates. Co2+ was inhibitory with both substrates, while Ca2+ had no significant effect. The phosphatase activities were inhibited by ATP, ADP, or AMP; the extents of inhibition were in opposite order with the two substrates. Phosphorylase phosphatase activity was strongly inhibited by KF or Pi. Phosphorylase phosphatase activity could be completely solubilized by incubating the membrane with 0.5 M NaCl or trypsin, and this was associated with several-fold activation. While Vmax values were increased, Km values for phosphorylase a were not much affected by these treatments. Unlike the soluble phosphatase, freezing in the presence of mercaptoethanol or by precipitation with ethanol failed to activate or to solubilize the membrane-bound phosphatase. The molecular weights of the NaCl-and the trypsin-solubilized phosphatase were estimated on gel filtration to be about 42,000 and 32,000, respectively. The present results indicate that the phosphoprotein phosphatase associated with liver plasma membrane shares several properties in common with phosphatases from other sources reported, and that, like those in the soluble fraction, it may be bound to some inhibitory proteins. PMID- 6270068 TI - Mode of hydrolysis of diribonucleoside monophosphates by phosphodiesterase phosphomonoesterase of Fusarium moniliforme. AB - The products derived from the degradation of the sixteen possible diribonucleoside monophosphates (NpN') by Fusarium phosphodiesterase phosphomonoesterase were analyzed by means of thin layer chromatography. The analysis showed that NpN' was first cleaved into nucleoside N and 5'-nucleotide pN', which was then dephosphorylated to yield nucleoside N'. The dephosphorylation was fast when N' was adenosine or cytidine but slow when N' was guanosine or uridine. The cleavage reaction was followed by measuring the increase of absorbance due to hyperchromicity, and the kinetic constants, Km and kcat, were determined for the sixteen dinucleoside phosphates. The Km value was higher, for a given N, when N' was a pyrimidine nucleoside than when N' was a purine nucleoside. For a given N', uridine as N gave the highest Km value and adenosine gave the lowest one. The kcat value was the highest, for a given N, when N' was cytidine. For a given N', uridine as N gave by far the lowest kcat value. These results can be interpreted in terms of two binding sites on the enzyme with different base preferences. Comparison of kcat/Km values suggested that the base of nucleoside N plays an important role in determining whether a dinucleoside phosphate is a good substrate of the enzyme. The dinucleoside phosphates with uridine as N were found to be particularly poor substrates of the enzyme. PMID- 6270069 TI - Regulation of electron transfer by sidedness-dependent surface pH. Dependence of the rate of cytochrome c-555 reduction on H+ concentration in the surface region on the periplasmic side of photosynthetic membranes in whole cells, spheroplasts and chromatophores of Chromatium vinosum. PMID- 6270070 TI - A spin-label study of protein-lipid interaction in sarcoplasmic reticulum of rabbit skeletal muscle. AB - Whether or not the thermotropic change at about 18 degrees C in the physical state of Ca2+-ATPase protein molecules of sarcoplasmic reticulum membranes could be transmitted to lipids through protein-lipid interactions was investigated using a spin-label technique. Fatty acid spin labels were used to probe the bulk membrane lipids while long-chain spin labels attached at one end to the Ca2+ ATPase molecules through a covalent bond were used to monitor the boundary lipids. The results on the temperature-dependence of alkyl-chain flexibility of lipid molecules indicate that the change in the state of the protein molecules is accompanied by one of the boundary lipids, but not of the bulk lipids. PMID- 6270071 TI - Transcriptional activity of globin genes in uninducible variants of Friend leukemic cells. AB - Transcriptional activity of globin genes in inducible and uninducible Friend leukemic cells was studied by two different approaches in an attempt to identify the biochemical basis for the loss of inducibility. (1) Nuclei were isolated, and transcripts synthesized endogenously were purified by the Hg-UTP/sulfhydryl Sepharose method. Globin mRNA sequences were quantitated by hybridization with [32P]globin cDNA. The results showed that the transcription of globin genes was stimulated in DMSO-treated inducible cell nuclei, whereas there was no activation of globin gene transcription in nuclei of uninducible variants cultured with DMSO. (2) Nuclei were prepared from inducible and uninducible Friend cells cultured in the presence or absence of DMSO, and were then subjected to limited digestion with DNase I. DNA was isolated from the nuclei after about 10% of total DNA had become acid-soluble, and was hybridized with globin cDNA. Globin genes either in inducible or in uninducible Friend cells were specifically susceptible to DNase I irrespective of the addition of inducer. From these studies, it was concluded that globin genes of uninducible Friend cells retained the "active conformation" but were not actively transcribed even in the presence of inducers. PMID- 6270072 TI - RNA polymerase of influenza virus. I. Comparison of the virion-associated RNA polymerase activity of various strains of influenza virus. AB - A systematic and comparative study was performed on the polypeptide composition and the RNA polymerase activity associated with virions of various strains of influenza A virus, including four human and two avian viruses. Significant differences were found in the molecular weights of not only hemagglutinin (HA) but also both nucleoprotein (NP) and membrane protein (M), as determined by polyacrylamide gel electrophoresis under denaturing conditions. The results indicate that, among viruses sharing the same serotype determined by the surface proteins HA and NA (neuraminidase), considerable variations exist in the structure of viral proteins, including inner proteins. The relative contents of viral proteins also varied among these strains grown under similar conditions. The total content of three P proteins, the putative RNA polymerase subunits, was within the range between 1.1 and 2.2% of total viral proteins and roughly paralleled the virion-associated RNA polymerase activity. The virion-associated RNA polymerase of all the strains tested were stimulated by the same dinucleotide primers, ApG or GpG, indicating that the specificity of transcription initiation is conserved among wide varieties of influenza virus. PMID- 6270073 TI - Sulfated oligosaccharides isolated from the deamination products of heparins. AB - Porcine heparin, whale heparin, and a solvolyzed porcine heparin were deaminated, and sulfated oligosaccharides, compounds 3f, 4f, 3s, 4s, 5, 6, 7s, 10, 11f, 11s, and 13 were isolated from the deamination products by Dowex 1 x 2 (Cl- form) column chromatography and high voltage paper electrophoresis and/or gel filtration on Sephadex G-25. Based on the results of chemical, 1H and 13C NMR spectral analyses, and of Smith degradation, together with previous observations, the structures of these sulfated oligosaccharides are proposed to be as follows: compound 3f, IdUA(2S)alpha 1 leads to GlcNAc alpha 1 leads to 4GlcUA; compound 4f, IdUA alpha 1 leads to 4GlcNAc(6S) alpha 1 leads to 4GlcUA; compound 3s, IdUA(2S) alpha 1 leads to 4GlcNAc alpha 1 leads to 4 GlcUA beta 1 leads to 4a Man; compound 4s, IdUA alpha 1 leads to 4Glc NAc(6S) alpha 1 leads to 4 GlcUA beta 1 leads to 4aMan; compound 5, IdUA(2S) alpha 1 leads to 4aMan; compound 6, GlcUA beta 1 leads to aMan(6S); compound 7s, IdUA alpha 1 leads to 4aMan(6S); compound 10, IdUA(2S)alpha 1 leads 4GlcNAc(6S)alpha 1 leads to 4 GlcUA beta 1 leads to 4aMan; compound 11f, IdUA(2S) alpha 1 leads 4GlcNAc alpha 1 leads to 4GlcUA beta 1 leads to 4a Man (6S); compound 11s, IdUA alpha 1 leads to GlcNAc(6S) alpha 1 leads to 4GlcUA beta 1 leads to 4aMan(6S); compound 13, IdUA(2S) alpha 1 leads to 4aMan(6S). For ths sulfated disaccharides, the same results as those reported in our previous papers were obtained. On the other hand, the proportion of total sulfated tri- and tetrasaccharides from whale heparin was 1.9 times higher than that from porcine heparine, reflecting a higher content of GlcNAc in the former. Also, the yields of compound 11s from these two heparins were comparable to their anticoagulant activities. In addition, certain 2-O-sulfates on IdUA flanked with GlcNS(6X) (X=H or S) in the heparin molecule are suggested to be important for the activity. PMID- 6270074 TI - Exposure of aromatic residues of Streptomyces subtilisin inhibitor. A photo-CIDNP study. AB - Exposure of aromatic residues, Tyr 7, Tyr 75, Tyr 93, His 43, His 106, and Trp 8, was studied by laser-induced photo-CIDNP in the 1H NMR spectrum of Streptomyces subtilisin inhibitor at 360 MHz. Only Tyr 7 and Tyr 75 gave strong CIDNP signals, whereas the rest of the aromatic residues gave no detectable signals in the temperature range 25-55 degrees C. From the temperature dependence data, it is concluded that Tyr 7 is well exposed at all temperatures, whereas the exposure of Tyr 75 increases with temperature, in agreement with the conclusion obtained by other methods. Agreements and discrepancies between the conclusions derived from the CIDNP data and the results so far obtained by other methods are compared for all the aforementioned aromatic residues. PMID- 6270075 TI - A transient spin-state change during alkaline isomerization of ferricytochrome c. AB - Kinetic difference spectra during the alkaline isomerization of ferricytochrome c were obtained by the pH-jump method in the range of 540 to 655 nm. The spectrum of the transient intermediate, which appears during the course of the isomerization, was reproduced from the spectra. The intermediate showed an intense absorption band at 600 nm, indicating that it is a high spin or mixed spin species. This is in contrast to the stable neutral and alkaline forms which are low spin species. The transient spin-state change during the isomerization was also observed upon rapid oxidation of ferrocytochrome c at alkaline pH. PMID- 6270076 TI - Nuclear location of ribonuclease H and increased level of magnesium-dependent ribonuclease H in rat liver on thioacetamide treatment. AB - Rat liver nuclei were isolated in aqueous solutions of low ionic strength or anhydrous glycerol. The presence of ribonuclease H (RNase H) [EC 3.1.4.34] activity in the cytoplasm is due to extraction of the nuclear enzyme by buffer and inorganic salts. Two forms of RNase H were separated from rat liver nuclei by affinity chromatography using a DNA-cellulose column. When the RNase H in the wash solution of nuclei with 0.3 M sucrose and in nuclear solution extracted with 0.15 M NaCl were fractionated on a single-stranded DNA-cellulose column, two peaks corresponding to Mn2+- and Mg2+-dependent RNases H were eluted at 0.1 M and 0.2 M NaCl, respectively, and a peak having both RNase H activities was recovered in the wash-through fraction from the column. Among the enzymes from these two fractions in the nuclei, the activity of the Mg2+-dependent RNase H which binds to DNA-cellulose increased several-fold within 24 h of a single injection of thioacetamide. The activities of Mg2+-dependent RNase H extracted with higher salt solution from the nuclei and recovered in the flow-through fraction from the DNA-cellulose column and the Mn2+-dependent RNase H activities were relatively unaffected by an injection of thioacetamide. PMID- 6270077 TI - Methanol dehydrogenase of Methylomonas J: purification, crystallization, and some properties. AB - A methanol dehydrogenase [EC 1.1.99.8] was purified and crystallized from methanol-grown Methylomonas J (formerly Pseudomonas sp. J), an obligate methylotroph. Its molecular weight was estimated to be 135,000 by gel chromatography. The sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis revealed two bands and their molecular weights were approximately 60,000 and 10,000. The enzyme was relatively stable at pH 6 and 10, and was unstable at pH 8. The enzyme activity was lost at pH 4.0; however, the prosthetic group was not liberated from the enzyme. Its isoelectric point was pH 9.3. The visible-ultraviolet absorption, fluorescence, CD, and ESR spectra were measured. The amino acid composition was analyzed after separation of the two components. Primary alcohols and formaldehyde served as substrates. Phenazine methosulfate (PMS) was an effective electron acceptor of the enzyme in the presence of either NH4Cl or methylamine. The enzyme was inhibited partially by metal chelators and completely by Mn2+ and Co2+. PMID- 6270078 TI - Two cytochromes c of Methylomonas J. AB - Two kinds of c-type cytochromes, cytochrome c-551 (I), and cytochrome c-551 (II), were highly purified and crystallized from cell-free extract of methanol-grown Methylomonas J (formerly Pseudomonas sp. J) and their physiochemical and biochemical properties were studied. Cytochrome c-551 (I) had an absorption peak at 409 nm in the oxidized form and peaks at 417, 523, 551 nm, and a shoulder at 532 nm in the reduced form. The millimolar extinction coefficient of the alpha peak of the reduced form was 25.3. The isoelectric point was at pH 5.3 and its standard redox potential was 0.29 V at pH 7.0. The molecular weight was estimated to be 16,000. Cytochrome c-551 (II) had absorption maxima at 409 nm in the oxidized form, and at 416, 521, and 551 nm in the reduced form. The millimolar extinction coefficient of the alpha-peak of the reduced form was 22.4. The isoelectric point was at pH 4.3 and its standard redox form was 22.4. The isoelectric point was at pH 4.3 and its standard redox potential was 0.24 V at pH 7.0. The molecular weight was estimated to be 12,500. The two cytochromes were reduced by methanol dehydrogenase [EC 1.1.99.8] of this bacterium, and formaldehyde was detected as an oxidation product. Ammonium chloride was not essential for reduction of the cytochromes. No significant reduction of the cytochromes was observed by methylamine dehydrogenase isolated from methylamine grown cells or by 2,6-dichlorophenol-indophenol (DCPIP)-dependent aldehyde dehydrogenase of the methanol-grown cells. The reduced forms of the cytochromes were oxidized by blue copper protein of the methanol-grown cells. PMID- 6270079 TI - Studies on the Ca2+-activated neutral proteinase of rabbit skeletal muscle. I. The characterization of the 80 K and the 30 K subunits. AB - The structure of the calcium-activated neutral proteinase (CANP) from rabbit skeletal muscle was examined. The purified CANP was homogeneous as judged by disc gel electrophoresis, while it showed two bands (M.W.=80,000 (80 K) and 31,000 (30 K) on SDS-gel electrophoresis. After denaturation of CANP, each subunit was separated and could be renatured alone or in combination with the other subunit. The results indicate that all the machinary necessary for the proteolytic activity is present in the 80 K subunit, but for the full activity the existence of the 30 K subunit is required. PMID- 6270080 TI - Limited autolysis of Ca2+-activated neutral protease (CANP) changes its sensitivity to Ca2+ ions. AB - Ca2+-activated neutral protease (CANP) usually requires mM Ca2+ for activation. The sensitivity of CANP to Ca2+ is greatly enhanced by passing it through a casein-Sepharose column in the presence of Ca2+ ions. This conversion is ascribed to autolysis of CANP. The converted enzyme required 40 microM Ca2+ for 50% activation. Various properties of the converted enzyme were very similar to those of CANP-I, recently found in canine heart muscle. Names of "m-CANP" and "mu-CANP" are proposed for CANPs which require mM and microM order Ca2+ for inactivation, respectively. PMID- 6270081 TI - Regulation of Ca2+-pumping ATPase of heart sarcolemma by a phosphorylation dephosphorylation Process. AB - The Ca2+-ATPase of dog heart sarcolemma (1, 2) is affected by phosphorylation. As normally prepared, sarcolemmal vesicles are phosphorylated to a high degree, resulting in a relatively low additional incorporation of hydroxylamine resistant [32P]phosphate from [gamma-32P]ATP. The 32P incorporation is increased up to 20 fold by pretreating the vesicles with phosphorylase phosphatase and is inhibited by an inhibitor of cAMP-dependent protein kinases. The phosphatase treatment inhibits markedly the Ca2+-ATPase and the ATP-dependent Ca2+ uptake. The inhibition is more evident at relatively higher levels of free Ca2+ and is reversed by preincubation with ATP. The Ca2+-pumping activity is stimulated markedly by phosphorylase b kinase and inhibited by the (cAMP-dependent) protein kinase inhibitor. Both the protein kinase inhibitor and ethylene glycol bis(beta aminoethyl ether)-N,N,N',N'-tetraacetic acid prevent the rephosphorylation of sarcolemmal vesicles, but the effects are not additive. The Ca2+ dependence curve of the Ca2+ uptake in phospho- and dephosphorylated vesicles suggests that the phosphorylation might affect the efficiency of the enzyme (turnover rate) rather than its affinity for Ca2+. PMID- 6270082 TI - The isolation and identification of 5,6-trans-25-hydroxyvitamin D3 from the plasma of rats dosed with vitamin D3. Evidence for a novel mechanism in the metabolism of vitamin D3. PMID- 6270083 TI - Thrombin-induced platelet responses differ in requirement for receptor occupancy. Evidence for tight coupling of occupancy and compartmentalized phosphatidic acid formation. PMID- 6270084 TI - Identification of a nerve growth factor receptor protein in sympathetic ganglia membranes by affinity labeling. AB - Membranes from adult rabbit superior cervical ganglia, cross-linked to membrane bound 125I-labeled nerve growth factor (NGF) by the photoreactive agent hydroxysuccinimidyl-p-azidobenzoate, were found to contain two labeled components with apparent Mr = 143,000 and Mr = 112,000. At high concentrations of the cross linker, minor amounts of a Mr = 300,000 affinity labeled product were also observed. The affinity labeled species exhibit the characteristics expected of membrane receptors for NGF. The inhibition of specific 125I-NGF binding to membranes by increasing concentrations of unlabeled NGF parallels the inhibiton of the affinity labeling of these components. Insulin, insulin-like growth factor I, multiplication stimulating activity, and epidermal growth factor do not inhibit the affinity labeling reaction. Membrane preparations of various non neuronal tissues do not show any detectable specific cross-linking to 125I-NGF. The affinity labeled species of superior cervical ganglia are proteins and contain intrapeptide disulfide bridges compacting their molecular structure. Peptide mapping experiments indicate a close structural relationship between the Mr = 143,000- and the Mr = 112,000-labeled proteins, suggesting a transformation of the former into the latter by limited proteolysis. The results suggest that the Mr = 143,000 affinity labeled protein represents a native NGF receptor component. PMID- 6270085 TI - Differential effects of alkylation of methionine residues on the activities of pituitary thyrotropin and lutropin. AB - Methionine residues of the alpha and beta subunits of bovine lutropin (LH) and bovine thyrotropin (TSH) have been specifically alkylated with iodoacetic acid. The alpha subunit has been modified so that two of the four methionines are quantitatively alkylated (residues 8 and 33, in agreement with studies by Cheng, K.-W. (1976) Biochem. J. 159, 71-77). Reassociation of the modified alpha subunit with unmodified LH-beta or thyrotropin (TSH)-beta resulted in reconstituted hormones which differed markedly in their respective biological activities. The alpha-modified TSH was fully active in both radioligand receptor and in vivo assays, while the alpha-modified LH, because of lowered affinity for receptor, lost approximately 70% of its activity in its radioligand receptor assay. This observation is the first to show that modification of the alpha subunit leads to a differential loss of activity in one glycoprotein hormone versus another. Circular dichorism studies revealed no changes in conformation; thus, the data strongly support, for LH, a direct interaction of the common subunit with receptor. Methionine 32 in TSH-beta can be modified with retention of full activity under conditions where methionines 8, 9, and 58 are not modified. In contrast, previous work on the modification of lysine 42 in LH-beta which lies in an analogous domain implicates that residue in receptor interaction (e.g. Liu, W. K., Yang, K.-P., Nakagawa, Y., and Ward, D. N. (1974) J. Biol. Chem. 249, 5544 5550; Sairam, M. R., and Li, C.-H., (1975) ARch. Biochem. Biophys. 167, 534-539). These results further emphasize the probable importance of this domain in hormone specificity. PMID- 6270086 TI - Ribonucleotide reductase in cultured mouse lymphoma cells. Cell cycle-dependent variation in the activity of subunit protein M2. AB - Ribonucleotide reductase is responsible for the production of the deoxyribonucleotides required for DNA synthesis. The enzyme is composed of two dissociable subunits, proteins M1 and M2, which are inactive alone, but are fully active when combined. From mouse S49 T lymphoma cells we have isolated and separated the two subunits and used each for determining the activity of the complementary subunit in extracts from cells of different phases in the cell cycle. Treatment of S49 cells with cAMP analogs (e.g. Bt2cAMP) results in the protein kinase-dependent arrest of the cells in the G1 phase of the cell cycle. Ribonucleotide reductase (holoenzyme) activity fell in S49 cells treated for more than 16 h with Bt2cAMP but was unchanged during short term treatments. The activity of protein M2 was decreased in parallel to the overall activity of ribonucleotide reductase, while protein M1 activity changed less. Removal of bt2cAMP after 24 h exposure resulted in increased holoenzyme and protein M2 activities. Centrifugal elutriation of exponentially growing S49 cells separated cells into a 90% pure G1 cell population a mixture of G1 and early S phase cells and a 95% pure S phase/G2 cell population. The specific catalytic activity of protein M1 was the same in all these fractions while that of protein M2 was decreased 60% in the G1 cell population. These results demonstrate that the ribonucleotide reduction necessary for DNA synthesis is regulated in a cell cycle dependent fashion by the activity of the protein M2 subunit of ribonucleotide reductase. PMID- 6270087 TI - Specific and saturable binding of plasma fibronectin to thrombin-stimulated human platelets. PMID- 6270088 TI - Multiple autopoly(ADP-ribosyl)ation of rat liver poly(ADP-ribose) synthetase. Mode of modification and properties of automodified synthetase. PMID- 6270089 TI - The collagen substrate specificity of human skin fibroblast collagenase. PMID- 6270090 TI - Human skin fibroblast collagenase. Assessment of activation energy and deuterium isotope effect with collagenous substrates. PMID- 6270091 TI - The reaction of sulfhydryl groups of sodium and potassium ion-activated adenosine triphosphatase with N-ethylmaleimide. The relationship between ligand-dependent alterations of nucleophilicity and enzymatic conformational states. AB - The reaction between N-ethylmaleimide and (Na+ + K+)-ATPase, performed under ligand conditions which produce each of the kinetic states of the enzyme and their associated conformational forms, was examined through an analysis of the inhibition of enzymatic activity and the incorporation of radiolabeled reagent into the enzyme. The inactivation reactions displayed pseudo-first order kinetics with respect to the concentration of active enzyme, indicating that the loss of activity is associated with the alkylation of a unique sulfhydryl group. In the absence of enzyme phosphorylation, the nucleophilicity of this sulfhydryl group is affected primarily by the nature of the monovalent cation present and does not correlate with the conformational state. A method for determining the actual concentration and specific radioactivity of radiolabeled N-ethylmaleimide during the reaction with (Na+ + K+)-ATPase was developed, allowing the measurement of the total reactive sulfhydryl groups of native (Na+ + K+)-ATPase under conditions identical with those of the inactivation studies. The labeling of the enzyme complex is associated almost exclusively with the large polypeptide, which contains four sulfhydryl groups which react with this reagent. One of these residues is presumably the sulfhydryl responsible for inactivation of the enzyme. Two react stoichiometrically and rapidly with N-ethylmaleimide under all conditions. The nucleophilicity of the fourth sulfhydryl group is governed by the conformational state of the enzyme, but the alkylation of this residue does not result in loss of enzymatic activity. PMID- 6270092 TI - Alteration in the protein components of catecholamine-sensitive adenylate cyclase during maturation of rat reticulocytes. AB - The maturing rat reticulocyte was used as a model system in which to study developmental changes in the protein components of hormone-sensitive adenylate cyclase. Plasma membranes from rat erythrocytes display 10 to 20% of the adenylate cyclase activity and 30 to 50% of the beta-adrenergic receptors which are measured in membranes from rat reticulocytes, as noted by others. Reticulocyte membranes also display equal activities in response to (-) isoproterenol in the presence of either GTP or GTP gamma S, whereas erythrocyte membrane adenylate cyclase is twice as active in the presence of isoproterenol plus GTP gamma S as in the presence of isoproterenol plus GTP. We have studied this system in greater detail by developing or applying independent assays for the catalytic protein (C) and the guanine nucleotide-binding regulatory protein (G/F) of adenylate cyclase. C was assayed in membranes by its intrinsic Mn2+ stimulated activity. It was also measured by reconstituting membranes with saturating amounts of GTP gamma S-activated G/F, yielding an operationally defined Vmax for the catalyst. By either assay, reticulocytes display about 3 fold greater C activity than do erythrocytes. G/F was assayed by its ability to confer GTP gamma S-stimulated activity upon C (which was supplied by membranes of cyc- S49 lymphoma cells). This assay indicates that reticulocyte membranes contain about 3 times as much G/F as do erythrocyte membranes. Cholera toxin and [32P]NAD were used to [32P]ADP-ribosylate the 45,000- and 52,000-dalton subunits of G/F. Total incorporation of 32P into these subunits decreased 3- to 4-fold with reticulocyte maturation. The ratio of label in the 52,000-dalton peptide to that in the 45,000-dalton peptide decreased from 0.29 in reticulocyte membranes to 0.14 in erythrocyte membranes. The apparently coordinate decrease in the amounts of C, G/F, and beta-adrenergic receptors suggest that the stoichiometry between these components is maintained during maturation, and may account for the decrease in adenylate cyclase in the membranes. However, the qualitative changes in responsiveness to hormones in the presence of GTP or GTP gamma S may be related to loss or proteolysis of the 52,000-dalton subunit of G/F. PMID- 6270093 TI - Equine follicle-stimulating hormone. Purification, acid dissociation, and binding to equine testicular tissue. AB - A simple method of purification of equine follicle-stimulating hormone is described by which two forms of the hormone are obtained. The acid dissociation of the most active preparation was studied and a pKa of 5.8 was determined at 37 degrees C. This value is 2 pH units higher than that observed for pregnant mare serum gonadotropin suggesting that the binding areas between subunits are not identical in the two hormones. We also describe an homologous radioreceptor assay of equine follicle-stimulating hormone which is highly specific for this hormone in contrast to the heterologous systems described so far. The analysis of the properties of equine gonadotropins in homologous and heterologous radioreceptor assays suggests that all glycoprotein hormones share a common high affinity binding site and that specificity of binding is due to binding prohibition sites preventing fixation of each hormone to the receptors of the other glycoprotein hormones. This specific hindering is defined as "negative specificity." PMID- 6270094 TI - Manifold effects of sodium butyrate on nuclear function. Selective and reversible inhibition of phosphorylation of histones H1 and H2A and impaired methylation of lysine and arginine residues in nuclear protein fractions. AB - In addition to its known effect in suppressing the deacetylation of the nucleosomal core histones, sodium butyrate in the concentration range 0.5 to 15 mM causes a selective inhibition of [32P]phosphate incorporation into histones H1 and H2A of cultured HeLa S3 cells. No commensurate general inhibition of phosphorylation is seen in the non-histone nuclear proteins of butyrate-treated cells, but phosphorylation patterns are altered and 32P-uptake may be stimulated, as well as inhibited, depending upon the protein fraction analyzed. The degree of inhibition of histone phosphorylation in intact cells increases progressively as the butyrate concentration is raised from 0.5 to 15 mM. The effect is time dependent and fully reversible. Butyrate has no effect on the kinetics of phosphate release from previously phosphorylated histones of cultured cells, nor does it significantly alter the rate of dephosphorylation of 32P-labeled histone H1 by endogenous phosphatases in vitro. Despite the suppression of [32P]phosphate incorporation into histones H1 and H2A of butyrate-treated cells, Na-butyrate does not inhibit the in vitro activities of either type I or type II cyclic AMP dependent protein kinases, or the cAMP-independent H1 kinase associated with cell cycle progression. This suggests that the butyrate effect on histone phosphorylation in vivo is indirect and may involve an alteration in substrate accessibility or a modulation of systems affecting kinase activities. The poly(ADP)-ribosylation of HeLa histones is not inhibited by 5 mM Na-butyrate. Cells exposed to butyrate show an impaired methylation of lysine and arginine residues in their histones and nuclear hnRNP particles, respectively. PMID- 6270095 TI - Cloning and structural characterization of an estrogen-dependent apolipoprotein gene. AB - ApoVLDLII is a major polypeptide component of avian very low density lipoprotein. Its production in the liver of the maturing female chick is developmentally synchronized with vitellogenesis. The apoVLDLII gene is normally dormant in males but can be activated by the administration of estrogen. In these studies, we describe the isolation of three recombinant bacteriophages that contain apoVLDLII genes. The genes appear to be identical from preliminary restriction analyses, although heterogeneity in flanking sequences is evident. The structure of the gene has been characterized and its organization correlated with the structure of the apoVLDLII polypeptide. PMID- 6270096 TI - Structure of the chicken apo very low density lipoprotein II gene. AB - We describe two cloned genomic DNA fragments, both bearing the entire apo very low density lipoprotein II gene. Electron microscopy and restriction enzyme mapping showed that this gene is split into at least four coding sequences by three or more intervening sequences. A very short exon at the 5'-end of the gene is separated by a 1.5-kilobase intron from the second exon, which codes for the AUG initiation codon of the mRNA. PMID- 6270097 TI - Structure and expression of human globin genes introduced into mouse fibroblasts. AB - The human delta- and beta-globin genes, contained in a recombinant bacteriophage (lambda H beta G1), were introduced into mouse fibroblasts by cotransformation with a plasmid (chi 1) containing the herpes simplex thymidine kinase gene using the calcium phosphate precipitation technique. A molar ratio of lambda H eta G1 to chi 1 DNA of 3:1 was used. Four of the eleven stable transformants obtained contained intact delta- and beta-globin genes as determined by Southern blot analysis. To assess methylation in the segment of human DNA introduced into mouse cells, digestion with Hpa II or Msp I alone or with a second restriction enzyme was performed. The sites examined near the human delta- and beta-globin genes in transformed cells were not methylated. RNA extracted from the transformed cells was analyzed by RNA-cDNA hybridization; no more than 100 copies of human beta globin mRNA/cell were found. Although hypomethylation of sites surrounding expressed globin genes in erythroid cells has been described, this property is not sufficient to ensure a high level of expression in fibroblasts. PMID- 6270098 TI - Ca2+ control of actin filament length. Effects of macrophage gelsolin on actin polymerization. AB - Gelsolin complexes with calcium (gelsolin-Ca2+) binds to the ends of actin filaments to which monomers add preferentially during elongation. It forms a stable complex with actin in a low ionic strength solution which does not normally favor the polymerization of actin. Gelsolin-Ca2+ increases the rate of nucleation of actin which precedes polymerization, but decreases the rate of elongation of the filaments. The final average length of filaments formed in the presence of gelsolin-Ca2+ is shorter and the equilibrium monomer concentration increases relative to actin polymerized in the absence of gelsolin-Ca2+. Gelsolin Ca2+ also increases the number of actin filaments because the magnitude of the increase in monomer concentration is disproportionately small compared with the reduction in polymer length. In these respects, the population of actin filaments formed during polymerization in the presence of gelsolin-Ca2+ is similar to that resulting from the action of gelsolin on previously assembled actin filaments (Yin, H. L., Zaner, K. S., and Stossel, T. P. (1980) J. Biol. Chem. 255, 9494 9500). The calcium-dependent shortening of ects, the population of actin filaments formed during polymerization in the presence of gelsolin-Ca2+ is similar to that resulting from the action of gelsolin on previously assembled actin filaments (Yin, H. L., Zaner, K. S., and Stossel, T. P. (1980) J. Biol. Chem. 255, 9494-9500). The calcium-dependent shortening of ects, the population of actin filaments formed during polymerization in the presence of gelsolin-Ca2+ is similar to that resulting from the action of gelsolin on previously assembled actin filaments (Yin, H. L., Zaner, K. S., and Stossel, T. P. (1980) J. Biol. Chem. 255, 9494-9500). The calcium-dependent shortening of actin filaments is the primary mechanism for the dissolution of an actin gel by gelsolin. Therefore, the ability of gelsolin to produce short filaments irrespective of the initial state of assembly of the actin offers flexibility for controlling the network structure of the cytoplasm in which either the monomeric or polymeric form of actin molecules might predominate at different times. PMID- 6270099 TI - beta-Adrenergic regulation of protein phosphorylation and its relationship to exocrine secretion in dispersed rat parotid gland acinar cells. AB - The possible role of cyclic AMP-mediated phosphorylation events in the regulation of exocrine secretion after beta-adrenergic stimulation was examined in vitro in dispersed acinar cell aggregates from rat parotid gland. l-Isoproterenol, a beta adrenergic agonist, stimulated endogenous activity of cyclic AMP-dependent protein kinase, alterations in the 32P content of 3 parotid phosphoproteins (increased 32P in 2, Mr = 27,000 and 14,000; decreased 32P in the remaining, Mr = 13,600), and amylase secretion in a dose-dependent manner. All responses were half-maximal within a range of l-isoproterenol concentrations of approximately 4 X 10(-8) to 5 X 10(-7) M. Examination of the time course of these 3 processes revealed that by 30 s after addition of l-isoproterenol, significant elevations in cyclic AMP-dependent protein kinase activity and alterations in the 32P content of the 3 parotid proteins had occurred, whereas secretion of amylase from cells was first detected 1-2 1/2 min after hormonal stimulation. Dibutyryl cyclic AMP (2 mM) elicited the same changes in parotid protein 32P content as l isoproterenol. Our results support the concept of a role for cyclic AMP-regulated protein phosphorylation in the sequence of cellular events leading to exocrine protein secretion from the rat parotid gland following beta-adrenergic stimulation. PMID- 6270100 TI - Purification of the putA gene product. A bifunctional membrane-bound protein from Salmonella typhimurium responsible for the two-step oxidation of proline to glutamate. AB - In this paper we report the purification of a protein which is able to catalyze both the proline oxidase and the pyrroline-5-carboxylic acid dehydrogenase activities necessary for the oxidation of proline to glutamic acid. The purification involves the preparation of a crude membrane pellet, detergent solubilization, ammonium sulfate fractionation, and DEAE-chromatography. We are able to obtain an essentially pure preparation (greater than 95% pure) after only a 52-fold purification, demonstrating that the protein is a major protein in cells fully induced for proline utilization. Both proline oxidase and pyrroline-5 carboxylic acid dehydrogenase activities co-purity throughout our purification. Velocity sedimentation of the purified protein demonstrates that both proline oxidase and pyrroline-5-carboxylic acid dehydrogenase activities co-sediment. Early in the purification procedure we are able to detect two species of protein which have both proline oxidase and pyrroline-5-carboxylic acid dehydrogenase activities. Our procedure purifies only the larger molecular weight species. The purified protein is a dimer composed of identical 132,000-dalton subunits. Analysis of mutants defective for proline utilization demonstrate that the bifunctional enzyme is the putA gene product. PMID- 6270101 TI - Enzymatic properties of the purified putA protein from Salmonella typhimurium. AB - In the previous paper (Menzel, R., and Roth, J. (1981) J. Biol. Chem. 256, 9755 9761) we have described the purification of a protein, the putA gene product, which has both proline oxidase and pyrroline-5-carboxylic acid dehydrogenase activities. In this paper we demonstrate that these enzyme activities are distinct with respect to a number of characteristics. The oxidase activity proceeds by a ping-pong mechanism involving the reduction of an enzyme-bound flavin. The dehydrogenase activity utilizes an ordered reaction mechanism. PMID- 6270102 TI - Pardaxin, a hydrophobic toxin of the Red Sea flatfish, disassembles the intact membrane of vesicular stomatitis virus. AB - Reaction of vesicular stomatitis virus with pardaxin, the hydrophobic toxin of the Red Sea flatfish, resulted in a profound morphological change of many virions and dissociation of their membrane and nucleocapsid into components readily separable by density gradient centrifugation. The basic matrix protein and acidic pardaxin segregated largely with the high density nucleocapsid. The dissociated virion membrane formed lipoprotein vesicles which retained glycoprotein spikes and a certain amount of N protein but no appreciable amounts of other nucleocapsid proteins and little if any RNA. Iodination of the tyrosine residue of the glycoprotein tail fragment provided supporting evidence that the COOH terminus of the glycoprotein extends beyond the inner layer of the membrane into the interior of the virion. These data indicate that pardaxin may serve as a probe for studying the organization of viral membranes, and, hopefully, other biological membranes. PMID- 6270103 TI - ATP-citrate lyase kinase and cyclic AMP-dependent protein kinase phosphorylate different sites on ATP-citrate lyase. AB - ATP-citrate lyase was phosphorylated by highly purified cyclic AMP-independent protein kinase (ATP-citrate lyase kinase) or the catalytic subunit of cyclic AMP dependent protein kinase. Each kinase phosphorylated ATP-citrate lyase equally but the combination of both kinases increased ATP-citrate lyase phosphorylation additively. When ATP-citrate lyase was phosphorylated with each kinase and partially digested with either L-1-tosylamido-2-phenylmethyl chloromethyl ketone treated trypsin or Staphylococcus aureus protease followed by electrophoresis of the proteolytic products on sodium dodecyl sulfate-polyacrylamide gels or when the phosphorylated lyase was completely digested by these proteases followed by chromatography and electrophoresis, the results showed that the site phosphorylated by ATP-citrate lyase kinase was different from that phosphorylated by the catalytic subunit of cyclic AMP-dependent protein kinase. Only phosphoserine was found in lyase phosphorylated by the catalytic subunit of cyclic AMP-dependent protein kinase whereas phosphoserine and phosphothreonine were found in ATP-citrate lyase phosphorylated by lyase kinase. PMID- 6270104 TI - Identification of inhibitor-2 as the ATP-mg-dependent protein phosphatase modulator. AB - In previous reports (Yang, S. D., Vandenheede, J. R., Goris, J., and Merlevede, W. (1980) J. Biol. Chem. 255, 11759-11767; Vandenheede, J. R., Yang, S.-D., and Merlevede, W. (1981) J. Biol. Chem. 256, 5894-5900), we have described two slightly different purification procedures for the isolation of the inactive ATP Mg-dependent protein phosphatase (FC) which could be activated by a protein activator (FA) in the presence of ATP-Mg. Although the two procedures consistently produced FC preparations that showed very similar polyacrylamide gel patterns, the specific activity of the purified enzymes varied considerably. The preparations characterized by a rather low specific activity could be stimulated up to 10-fold when a titrated amount of inhibitor-2 was included during the FA and ATP-Mg-mediated activation. Inhibitor-2, only recently implicated as an inactivating protein for activated FC (Vandenheede, J. R., Goris, J., Yang, S. D., Camps, T., and Merlevede, W. (1981) FEBS Lett. 127, 1-3) has now been identified as a modulator protein, necessary for the reversible activation of the protein phosphatase. PMID- 6270105 TI - Cytochrome c oxidase in cytochrome c oxidase-deficient mutant strains of Neurospora crassa. AB - Three mutant strains of Neurospora crassa, previously characterized as cytochrome c oxidase deficient, were found to possess this respiratory enzyme. The mitochondrial cytochrome contents of logarithmic phase cells of wild type (74A) and the respiration-deficient strains mi-3, cya-4-23, and cya-5-34 were investigated with low temperature difference spectrophotometry; two of the strains were found to possess cytochrome a. Cytochrome c oxidase catalytic activity measurements performed on isolated mitochondria showed that all three mutant strains contained significant amounts of active enzyme which was subsequently found to be active in vivo. However, activity of the cyanide insensitive, alternate electron transport pathway was responsible for most of the oxygen consumed by the logarithmic phase cells of these mutant strains. Electrophoretic analysis of the polypeptide subunits of cytochrome c oxidase radiolabeled in vivo during logarithmic phase growth showed that, in contrast to previous reports, the respiration-deficient strains appeared to contain all seven subunits of the enzyme. PMID- 6270106 TI - Studies on cobalt myoglobins and hemoglobins. The interaction of molecular oxygen with leghemoglobin. AB - Cobalt-substituted leghemoglobin (CoLb) was prepared from cobaltous protoporphyrin IX and apo-leghemoglobin a, and its functional and spectroscopic properties were examined. The oxygen affinity of CoLb was pH-dependent. The partial oxygen pressure at half-saturation was decreased from 14.3 torr at pH 8 to 3.5 Torr at pH 3.5 with a pK value of 5.8 at 15 degrees C. The oxygen association and dissociation rates were determined by the temperature jump relaxation technique. The association rate was essentially pH-independent, and the relatively high oxygen affinity of CoLb was found to be due to the fast oxygen association rate in comparison with cobalt myoglobin. The oxygen dissociation rate was pH-dependent with a pK value of 5.7 at 15 degrees C, suggesting that the pH dependence of the oxygen dissociation rate is responsible for the pH-dependent change in the oxygen affinity of CoLb. The EPR spectrum of oxyCoLb exhibited a pH-dependent change with a pK value of 5.7 at 15 degrees C. The EPR spectrum at 77 K of oxyCoLb at neutral pH exhibited a small effect upon the replacement of H2O with D2O, but it was drastically altered in the acidic pH region. The deoxy EPR spectrum was pH-independent between pH 4 and 9. These observations indicate that in oxyCoLb, the bound oxygen is interacting with the distal amino acid group with a pK value of 5.7 and that this interaction is responsible for the increased oxygen affinity in the acidic pH region. PMID- 6270107 TI - Mitochondrial respiratory capacity and Na+- and K+-dependent adenosine triphosphatase-mediated ion transport in the intact renal cell. AB - Cellular oxygen consumption was monitored during stimulation and inhibition of the Na+- and K+-dependent adenosine triphosphatase in a suspension of intact tubules isolated from the rabbit renal cortex. Respiratory rates were compared to the ADP-stimulated respiratory rate (state 3 rate) obtained in mitochondria released directly from the renal tubules by digitonin shock. At 37 degrees C, in the presence of NADH-linked substrates and fats, isolated renal cells respire at 50 to 60% of the state 3 rate. Inhibition of the (Na+,K+)-ATPase with the cardiac glycoside, ouabain, results in a decline in respiration to 25 to 30% of the state 3 rate. Stimulation of the (Na+,K+)-ATPase produced as a result of nystatin mediated dissipation of plasma membrane Na+ and K+ gradients results in increased respiration with an oxygen consumption rate characteristic of optimal ATP synthesis (state 3). The relationship between metabolic substrate regimen, mitochondrial respiratory capacity, and cellular energy demand is examined in the context of these findings. PMID- 6270108 TI - Purification and properties of a topoisomerase from Ustilago maydis. AB - The lower eukaryote Ustilago maydis contains a topoisomerase that removes supercoils from negative and positive superhelical DNA. The enzyme may be a multimeric protein or an aggregate of polypeptides with a native molecular weight of 270,000 as estimated by gel filtration. No cofactors are required by the enzyme, but activity is enhanced by Mg2+. Dependence of activity upon enzyme concentration is not linear. Below a threshold level where topoisomerase cannot ordinarily be detected, addition of H1 histone sharply stimulates activity. ATP and a number of structural analogues inhibit the enzyme. PMID- 6270109 TI - Purine amplification of luteinizing hormone action in ovarian luteal cells. PMID- 6270110 TI - Pituitary gastrins. Different processing in corticotrophs and melanotrophs. AB - The anterior, intermediate, and neural lobes of porcine pituitaries contain different molecular forms of gastrin. In the anterior lobe, unsulfated large gastrins, component I, and gastrin-34 constitute the main forms. In contrast in the intermediate and neural lobes, gastrin-17, sulfated as well as unsulfated, predominates. Since component I and gastrin-34 are biosynthetic precursors of gastrin-17, the findings indicate that proteolytic processing and amino acid derivatization (sulfation) differs between the lobes. This difference, together with the localization of anterior lobe gastrin in corticotrophs and intermediate lobe gastrin in melanotrophs, emphasizes the close relation and parallelism of the biosynthesis of gastrin and corticotropin peptides. PMID- 6270111 TI - Uptake and 25-hydroxylation of vitamin D3 by isolated rat liver cells. AB - The physiological roles played by hepatocytes and nonparenchymal cells of rat liver in the metabolism of vitamin D3 have been investigated. Tritium-labeled vitamin D3 dissolved in ethanol was administered intravenously to two rats. Isolation of the liver cells 30 and 70 min after the injection showed that vitamin D3 had been taken up both by the hepatocytes and by the nonparenchymal liver cells. The relative proportion of vitamin D3 that accumulated in the nonparenchymal cells increased with time. Perfusion of the isolated rat liver with [3H] vitamin D3 added to the perfusate confirmed the ability of both cell types to efficiently take up vitamin D3 from the circulation. By a method based on high pressure liquid chromatography and isotope dilution-mass fragmentography it was found that isolated liver cells in suspension had a considerable capacity to take up vitamin D3 from the medium. About 2.5 fmol of vitamin D3 were found to be associated with each hepatocyte or nonparenchymal cell after 1 h of incubation. 25-Hydroxylation in vitro was found to be carried out only by the hepatocytes. The rate of hydroxylation was about the same whether the cells were isolated from normal or rachitic rats (3.5 and 4 pmol of 25-hydroxyvitamin D3 formed per h per 10(6) cells, respectively). The possibility that the nonparenchymal cells might serve as a storage site for vitamin D3 in the liver is discussed. PMID- 6270112 TI - Membrane transfer of alpha-tocopherol. Influence of soluble alpha-tocopherol binding factors from the liver, lung, heart, and brain of the rat. PMID- 6270113 TI - Fluorescence energy transfer studies of the interaction between adrenodoxin and cytochrome c. PMID- 6270114 TI - Analysis of 5' flanking sequences and intron-exon boundaries of the rat prolactin gene. PMID- 6270115 TI - Induction of phosphorylation of dolichol during embryonic development of the sea urchin. PMID- 6270116 TI - The respiratory chain of Bacillus alcalophilus and its nonalkalophilic mutant derivative. PMID- 6270117 TI - Identification of a renal receptor for parathyroid hormone by photoaffinity radiolabeling using a synthetic analogue. PMID- 6270118 TI - LDHk, a uniquely regulated cryptic lactate dehydrogenase associated with transformation by the Kirsten sarcoma virus. AB - A novel isozyme of lactate dehydrogenase is detected in various cells transformed by the Kirsten murine sarcoma virus (KiMSV). This isozyme, designated LDHk, is strongly inhibited by physiological concentrations of oxygen, in an apparently cooperative fashion. LDHk is inhibited by guanosine triphosphate and related compounds, in a noncompetitive fashion. LDHk is found with both 35,000- and 22,000-dalton subunits, although these probably cleave from a 57,000-dalton precursor. In studies utilizing a temperature-sensitive transforming gene mutant of the Kirsten sarcoma virus, we find in vivo expression of LDHk is also temperature-sensitive. In studies using either crude cell-free extracts or purified LDHk, we find the enzyme from cells infected with a temperature sensitive transforming gene mutant of KiMSV is thermolabile relative to that from wild type KiMSV-infected cells. PMID- 6270119 TI - Thyroglobulin interactions with thyroid membranes. Relationship between receptor recognition of N-acetylglucosamine residues and the iodine content of thyroglobulin preparations. AB - Bovine thyroglobulin has been subjected to sequential glycohydrolase treatment in order to define further the components of the carbohydrate chain which are important in binding of the glycoprotein to bovine thyroid membranes. Preparations of asialoagalactothyroglobulin exhibit the best binding, suggesting that exposed N-acetylglucosamine residues on the B carbohydrate chain of thyroglobulin play an important role in the interaction of thyroglobulin with the thyroid membranes. Enhanced binding of asialoagalactothyroglobulin to microsomal, lysosomal, and Golgi membranes, as well as to thyroid cells in culture, was also observed. Isopycnic rubidium chloride gradient centrifugation, a procedure used in the isolation of thyroglobulin molecules with a low iodine content, also isolates thyroglobulin molecules with a low sialic acid content and with an increased ability to interact with wheat germ agglutinin, a lectin which recognizes exposed N-acetylglucosamine residues. The studies further indicate that there is a correlation between iodine content, exposed N-acetylglucosamine residues, and the binding of thyroglobulin to thyroid membranes. PMID- 6270120 TI - Binding of thyroglobulin to bovine thyroid membranes. Role of specific amino acids in receptor recognition. AB - Bovine thyroglobulin was treated with increasing ratios of succinic anhydride, trinitrobenzene sulfonic acid, tetranitromethane, and N-acetylimidazole in an attempt to assess the role of lysine or tyrosine residues in binding to thyroid membrane receptors. Extensive succinylation results in dissociation to 12 S thyroglobulin with retention of a considerable portion of the three-dimensional structure. Only 25% of the lysine residues can be modified by trinitrophenylation without affecting inter-subunit interactions. Succinylation as well as trinitrophenylation increases the affinity of thyroglobulin for the membrane receptor by a factor of 2. The binding of thyroglobulin to the membrane was reduced after nitration of 30% of the tyrosyl residues with tetranitromethane. O Acetylation of 40-70% of the tyrosyl residues by N-acetylimidazole nearly abolished the ability of thyroglobulin to bind to the membrane. Removal of the O acetyl group with hydroxylamine restored the binding properties. The results indicate that tyrosyl residues play an important role in thyroglobulin interactions with thyroid membranes. PMID- 6270121 TI - Nucleotide assignment of alkali-sensitive sites in mouse mitochondrial DNA. AB - Mature, closed circular mouse mitochondrial DNA contains a significant number of ribonucleotides throughout the genome. Previous studies have implicated the two origins of DNA replication as preferred sites of ribonucleotide retention. We have analyzed the site specificity of ribosubstitution by direct sizing of alkali treated restriction fragments in comparison with the DNA sequence of untreated restriction fragments of cloned mouse mitochondrial DNA. These results have confirmed the observations that ribonucleotides are retained at the two origins of replication and are most likely remnants of RNA priming events associated with DNA replication. The map location of ribonucleotides at the light strand origin of replication has been refined to a triplet nucleotide (5'-CGG-3') in the light strand initiation region. This approach has demonstrated that all four deoxyribonucleotides are subject to ribosubstitution and no single base (or subset of the four bases) predominates. An examination of selected regions of the mitochondrial DNA genome including the putative coding region for cytochrome oxidase subunit III and regions containing the genes for tRNAPhe, tRNAVal, 12 S rRNA, and 16 S rRNA reveals preferred sites for ribosubstitution. These preferred sites do not relate in any obvious way to the functional aspects of these domains. In addition, the data indicate that every position in the DNA sequences examined can be ribosubstituted at a very low frequency. PMID- 6270122 TI - The mannose 6-phosphate receptor of Chinese Hamster ovary cells. Isolation of mutants with altered receptors. PMID- 6270123 TI - The mannose 6-phosphate receptor of Chinese hamster ovary cells. Compartmentalization of acid hydrolases in mutants with altered receptors. AB - The localization of acid hydrolases was examined in Chinese hamster ovary cells with defective mannose 6-phosphate receptors; these mutants had been shown to exhibit reduced uptake and altered binding of exogenously added acid hydrolase (Robbins, A. R., Myerowitz, R., Youle, R. J., Murray, G. J., and Neville, D. M., Jr. (1981) J. Biol. Chem. 256, 10618-10622). Cells were grown in the presence of [3H]mannose, alpha-L-iduronidase and beta-hexosaminidase were immunoprecipitated sequentially, electrophoresed on polyacrylamide gels containing sodium dodecyl sulfate, and detected by fluorography. About 55% of the alpha-L-iduronidase and beta-hexosaminidase synthesized by the mutants in 12 h was found in the growth medium; parental cells secreted only approximately 15%. The mutants also secreted 2 to 6 times more alpha-mannosidase, beta-glucuronidase, and alpha-L-fucosidase than the parent as determined by measurements of enzyme activity. Intracellular levels of these enzymes were reduced in the mutants. The mutants secreted acid hydrolases in the precursor forms, within the cells these enzymes resided in lysosomes and were processed normally; thus, the mutants appeared aberrant only with respect to distribution of hydrolases between intracellular and extracellular compartments. [35S]methionine-labeled beta-hexosaminidase and alpha L-iduronidase secreted by the mutants were taken up normally by both human fibroblasts and wild type CHO cells, and this uptake was inhibited by mannose 6 phosphate. Thus, the elevated secretion of acid hydrolases was not due to alteration of the mannose 6-phosphate recognition marker on the enzymes, but appears to result from alterations in the mannose 6-phosphate receptor. PMID- 6270124 TI - The regulatory role of cyclic AMP in hormone-induced of granulosa cell differentiation. PMID- 6270125 TI - Expression of the Streptomyces enzyme endoglycosidase H in Escherichia coli. AB - Endoglycosidase H is one of a large number of enzymes secreted by Streptomyces plicatus and other Streptomyces species. When the structural gene for this enzyme is introduced into Escherichia coli attached to the plasmid pBR-322 or Charon 4 phage, the enzyme is synthesized and is found in the periplasmic space, culture medium, and cells. Attachment of the UV-5 lac promoter to a site in the plasmid adjacent to the Streptomyces insert stimulates enzyme synthesis as much as 100 fold. This result demonstrates that transcription of the Streptomyces gene can be initiated from sequences outside of the Streptomyces insert. Initiation of transcription on a Streptomyces promoter is also a suggested but unproven possibility. In contrast to the situation in Streptomyces, where the enzyme has a molecular weight of 27,000, the enzyme made in E. coli has a molecular weight of approximately 30,000. Possible explanations for this difference in size are lack of cleavage of the Streptomyces secretion "signal sequence" in E. coli or protein "processing" by enzymes secreted into the medium by STreptomyces. PMID- 6270126 TI - The dnaB protein of Escherichia coli groPB mutants. PMID- 6270127 TI - Turnover of regulatory subunit of cyclic AMP-dependent protein kinase in S49 mouse lymphoma cells. Regulation by catalytic subunit and analogs of cyclic AMP. AB - Turnover of regulatory subunit (R) of type I cAMP-dependent protein kinase in intact S49 mouse lymphoma cells was studied using two-dimensional gel electrophoresis to analyze [35S]methionine label in R during label-chase experiments. R decays exponentially with a half-life of about 8.4 h in drug-free, wild type cells. In mutant cells lacking functional kinase catalytic subunit, R is about 10 times more labile than in wild type cells. 8-bromo-cAMP, isoproterenol, and cholera toxin destabilize R in wild type cells to an extent comparable to the "kinase-negative" mutation. In contrast, dibutyryl-cAMP stabilizes R in both wild type and kinase-negative cells. Sodium butyrate has no significant effect on R stability. These results are discussed in terms of R structure and the regulation of R expression. PMID- 6270128 TI - In vitro transcription of the histidine operon. Identification of the his promoter and leader and readthrough transcripts. PMID- 6270129 TI - A monoisozymic nucleoside diphosphate kinase capable of complete phosphorylation. AB - Nucleoside diphosphate kinase (EC 2.7.4.6) has been purified nearly 2000-fold from beef brain particulate material in 65% yield. The purification procedure described here takes advantage of the relatively unique complexation of the enzyme with blue dextran-Sepharose in the presence of a nonionic detergent. Unlike preparations of the enzyme previously described, the bovine brain enzyme is monoisozymic and each polypeptide chain can be phosphorylated by a nucleoside triphosphate substrate. Catalytically functional forms of the enzyme having lower pI values can be generated by proteolysis. PMID- 6270130 TI - EPR studies of the Mn(II) complex with elongation factor Tu and GDP Identification of oxygen ligands to Mn(II) by observation of 17O superhyperfine coupling. AB - The coordination sphere of Mn(II) in the complex with GDP and elongation factor Tu from Escherichia coli has been probed by EPR spectroscopy with 17O-labeled ligands. Inhomogeneous broadening in the EPR signals for Mn(II) due to unresolved superhyperfine coupling to the 17O nucleus was used to identify directly bound oxygen ligands. Results with GDP selectively enriched with 17O either in the alpha-phosphate or in the beta-phosphate revealed that GDP was a beta-monodentate ligand for Mn(II) in the complex with the protein. Results with 17O-enriched water showed that two water molecules are coordinated to the Mn(II). The EPR spectrum for the complex is characteristic of octahedral coordination for Mn(II). Hence, three ligands from the protein are required to complete the sextet of ligands. PMID- 6270131 TI - Effect of vitamin D status on cyclic AMP-dependent protein kinase activity and its heat-stable inhibitor in chick kidney. AB - Cyclic AMP-dependent protein kinase activity in supernatants of homogenates of kidneys from vitamin D-deficient chicks is decreased to 70% of the level measured in kidneys from normal chicks. Activity was restored to normal by oral administration of vitamin D or 1,25-dihydroxyvitamin D3 for 1 or 2 weeks. Both isozymes of cAMP-dependent protein kinase were reduced to the same extent by vitamin D deficiency. The decreased enzyme activity could not be accounted for by a shift to the particulate fraction nor by an increased requirement for cyclic AMP. A heat stable, trichloroacetic acid-precipitable, trypsin-labile inhibitor of protein kinase activity was identified and quantitated in kidneys from vitamin D-deficient chicks (16 to 26 units/mg of protein) and from those given vitamin D (2 to 6 units/mg of protein). The measured difference in inhibitor levels could not be attributed to differential stability in kidney homogenates from vitamin D deficient or -repleted chicks. The observed increase in inhibitor level with vitamin D deficiency is not sufficient to account for the decrease in cyclic AMP dependent protein kinase activity, suggesting that the total amount of this enzyme activity is reduced in vitamin D deficiency. PMID- 6270132 TI - Affinity labeling of cAMP-dependent protein kinase with p-fluorosulfonylbenzoyl adenosine. Covalent modification of lysine 71. AB - p-Fluorosulfonylbenzoyl 5'-adenosine (FSO2BzAdo) was shown previously to be an irreversible inhibitor of the catalytic subunit of cAMP-dependent protein kinase II from porcine skeletal muscle (Zoller, M. J., and Taylor, S. S. (1979) J. Biol. Chem. 254, 8363-8368). The catalytic subunit of porcine heart cAMP-dependent protein kinase was also inhibited following incubation with FSO2[14C]BzAdo, and inhibition was shown to result from the stoichiometric, covalent modification of a single lysine residue. The amino acid sequence in an extended region around the carboxybenzenesulfonyl lysine (CBS-lysine) was elucidated by characterizing both tryptic and cyanogen bromide peptides containing the 14C-modified residue. The sequence in this region was Leu-Val-Lys-His-Lys-Glu-Thr-Gly-Asn-His-Phe-Ala-Met Lys(CBS)-Ile-Leu-Asp-Lys-Glu-Lys-Val-Val-Lys-Leu-Lys-Gln-Ile. The covalently modified residue corresponded to lysine 71 in the overall polypeptide chain. Homologies to bovine heart catalytic subunit and to a site modified by FSO2BzAdo in phosphofructokinase are considered. PMID- 6270133 TI - Structure of the lutropin receptor on granulosa cells. Photoaffinity labeling with the alpha subunit in human choriogonadotropin. AB - Ovarian granulosa cells have specific receptors for lutropin and human choriogonadotropin (hCG). To investigate the nature of the receptor, several different photo-cross-linkable derivatives of radioiodinated hCG were incubated with porcine granulosa cells and hormone derivative-receptor complexes extracted with 0.5% Triton X-100. The extracted complexes appeared in a single peak on gel permeations and in rat zonal sedimentation studies. The estimated molecular weight was found to be 250,000-370,000 with an associated s value of 7.7 +/- 0.3. When Triton X-100 extracts were examined by gel electrophoresis, the autoradiograph revealed four bands. Three were conspicuous, corresponding for molecular weights of 96,000 +/- 5,000, 76,000 +/- 4,000, and 73,000 +/- 4,000, but a fourth band, corresponding to a molecular weight of 120,000 +/- 6,000, was less well defined. Reducing agents, 20 mM dithiothreitol and 2% beta mercaptoethanol, acid, and 3 M urea did not affect the production of the bands. Furthermore, the bands once formed were resistant to these agents, indicating 1) covalent nondisulfide bond formation in photoaffinity labeling and 2) noncovalent association of photoaffinity-labeled membrane components with the natural hormone receptor. PMID- 6270134 TI - Cyclic AMP-dependent protein kinase in mitochondria and cytosol from different sized follicles and corpora lutea of porcine ovaries. AB - cAMP-dependent protein kinase was examined in mitochondria and cytosol prepared from different-sized antral follicles and corpora lutea of porcine ovaries. In all ovarian tissues examined except small follicles, protein kinase-specific activity was significantly higher in mitochondria than in cytosol, with the highest to lowest activities being found in medium (4-6 mm) follicles, large (7 12 mm) follicles, corpora lutea, and small (1-3 mm) follicles, respectively. Using the photoaffinity analogue [32P]8-N3cAMP, two major cAMP binding proteins with Mr = 47,000 (the apparent regulatory subunit of protein kinase Type I) and 54,000-56,000 (Type II) were found in all ovarian preparations. Type II was predominant in the cytosol of all ovarian samples, with the cytosolic Type I to Type II ratio increasing from approximately 0.05 in small and medium follicles top approximately 0.20 in large follicles and corpora lutea. In contrast, ovarian mitochondrial preparations contained relatively more Type I than did cytosol, with the mitochondrial Type I to Type II ratio increasing from approximately 0.50 in small and medium follicles to 0.88 in large follicles and 2.96 in corpora lutea. Also, mitochondrial [4-14C]cholesterol conversion and 3 beta hydroxysteroid dehydrogenase/isomerase activities increased with follicle size and luteinization. These results suggest that Type I may play a role in the regulation of ovarian mitochondrial steroidogenesis. PMID- 6270135 TI - Specificity in the activation and inhibition by flavonoids of benzo[a]pyrene hydroxylation by cytochrome P-450 isozymes from rabbit liver microsomes. AB - The effect of flavone and 7,8-benzoflavone on the metabolism of benzo[a]pyrene to fluorescent phenols by five cytochrome P-450 isozymes obtained from rabbit liver microsomes was determined. Benzo[a]pyrene metabolism was stimulated more than 5 fold by the addition of 600 microM flavone to a reconstituted monooxygenase system consisting of NADPH, cytochrome P-450 reductase, dilauroylphosphatidylcholine, and cytochrome P-450LM3c or cytochrome P-450LM4. In contrast, an inhibitory effect of flavone on benzo[a]pyrene metabolism was observed when cytochrome P-450LM2, cytochrome P-450LM3b, or cytochrome P-450LM6 was used in the reconstituted system. 7,8-Benzoflavone (50-100 microM) stimulated benzo[a]pyrene metabolism by the reconstituted monooxygenase system about 10-fold when cytochrome P-450LM3c was used, but benzo[a]pyrene hydroxylation was strongly inhibited when 7,8-benzoflavone was added to the cytochrome P-450LM6-dependent system. Smaller effects of 7,8-benzoflavone were observed on the metabolism of benzo[a]pyrene by the cytochrome P-450LM2-, cytochrome P-450LM3b-, and cytochrome P-450LM4-dependent monooxygenase systems. These results demonstrate that the activating and inhibiting effects of flavone and 7,8-benzoflavone on benzo[a]pyrene metabolism depend on the type of cytochrome P-450 used in the reconstituted monooxygenase system. PMID- 6270136 TI - Generation of free radicals induced by nifurtimox in mammalian tissues. AB - Nifurtimox is reduced by rat liver microsomes to a nitro anion-free radical as indicated by ESR spectroscopy. This subcellular fraction gives a steady state radical concentration which is proportional to the square root of the protein concentration, suggesting that the nifurtimox anion radical is a necessary intermediate in the reduction and that the radical decays through a nonenzymatic second order process. The steady state concentration of the anion radical in the microsomal system is not decreased by superoxide dismutase or catalase, thus indicating that neither the superoxide anion nor hydrogen peroxide is an intermediary in the generation of the anion radical. The steady state concentration of the anion radical in the microsomal system is also not altered in the presence of metyrapone or CO and is decreased in the presence of NADP+ and p-chloromercuribenzoate. This observation suggests that the formation of nifurtimox anion radical is mediated through NADPH-cytochrome P-450 (c) reductase and not by the cytochrome P-450 system. In accordance with this interpretation, a model system consisting of NADPH and FMN-reduced nifurtimox to the nitro anion free radical. Nifurtimox anion radical generation is significantly stimulated by rat brain and testes homogenates. The enhanced free radical formation may be the basic cause of nifurtimox toxicity in mammals. PMID- 6270137 TI - 3,4-Dihydroxyphenylacetate 2,3-dioxygenase. A manganese(II) dioxygenase from Bacillus brevis. AB - 3,4-Dihydroxyphenylacetate 2,3-dioxygenase, an enzyme which catalyzes the extradiol cleavage of catechols, has been purified from Bacillus brevis. Like other extradiol-cleaving dioxygenases, this enzyme has a molecular weight of 140,000 with four subunits of 36,000 each. Unlike the other enzymes, this dioxygenase is not activated by added ferrous ion, not inhibited by cyanide or diethyldithiocarbamate, and not inactivated by H2O2. X-ray fluorescence and atomic absorption analyses show the enzyme to contain approximately 2 g atoms of manganese per mol of protein. EPR spectra are consistent with a manganese(II) center in an environment of low symmetry. This is the first report of an oxygen activating manganese enzyme. PMID- 6270138 TI - Stimulation of phosphatidic acid of calcium influx and cyclic GMP synthesis in neuroblastoma cells. AB - Phosphatidic acid added to the medium markedly elevated intracellular cyclic GMP content in cultured neuroblastoma N1E 115 cells. There was a significant elevation of cyclic GMP with 1 micrograms/ml and a maximum (70-fold) elevation with 100 micrograms/ml of phosphatidic acid. Other natural phospholipids did not increase, or increased only slightly, the cyclic GMP content in the cells. The elevation of cyclic GMP content by phosphatidic acid was absolutely dependent on extracellular calcium. Phosphatidic acid stimulated the influx of calcium into neuroblastoma cells 2- to 5-fold. The pattern of the calcium influx induced by phosphatidic acid was comparable to that of cyclic GMP elevation. The stimulation of calcium influx by phosphatidic acid was also observed in cultured heart cells, indicating that phosphatidic acid acts as a calcium ionophore or opens a specific calcium-gate in a variety of cell membranes. Treatment of neuroblastoma cells with phospholipase C increased 32Pi labeling of phosphatidic acid, stimulated the influx of calcium, and elevated the cyclic GMP content in the cells. Thus exogenous as well as endogenous phosphatidic acid stimulates the translocation of calcium across cell membranes and, as a consequence, induces the synthesis of cyclic GMP in the neuroblastoma cells. PMID- 6270139 TI - Phospholipid accumulation during the cell cycle in synchronous cultures of the yeast, Saccharomyces cerevisiae. AB - Phospholipid concentrations have been examined throughout successive cell cycles in synchronously growing cultures of the yeast, Saccharomyces cerevisiae. Total phospholipid phosphorus, as well as lecithin and phosphatidylethanolamine levels, exhibited stepwise increases during the cell cycle with step increments beginning just prior to new rounds of bud formation. Phosphatidylinositol and phosphatidylserine levels, on the other hand, showed what have been interpreted to be peak concentrations near the time of bud formation. Cardiolipin content varied considerably and was dependent upon the carbon source of the growth medium. Glucose-grown cells exhibited peak concentrations of cardiolipin near the time of bud formation, with marked decreases after this time. In contrast, galactose-grown synchronous cells exhibited stepwise increments in cardiolipin content, with step increases occurring near the time of new rounds of bud formation. Step or peak increases in cardiolipin, as well as all other phospholipids, were found to coincide with the time of stepwise increases in cytochrome c oxidase activity in these cells. No correlations were observed between the elaboration of mitochondrial membranes during the synchronous cell cycle and the observed patterns of phospholipid increase. PMID- 6270140 TI - Protein dye affinity chromatography using immobilized tetraiodofluorescein. AB - The chromatographic behavior of a heterogeneous protein mixture and of a series of homogeneous proteins on the immobilized dye tetraiodofluorescein has been observed and analyzed. Less than 6%, of the millimolar concentration of dye immobilized to a porous agarose matrix is accessible to protein. The affinity of a protein for immobilized dye is dramatically increased by insertion of apolar spacer atoms between the dye and the matrix. Dye columns constructed with a 9 atom spacer can be used to advantage for the retention and competitive elution of proteins not found previously amenable to dye chromatography. PMID- 6270141 TI - Large scale isolation and properties of subunits from bovine heart cytochrome oxidase. AB - The subunits of the cytochrome oxidase from bovine heart were isolated in large quantities suitable for amino acid sequence studies. The preparation of subunits III, IV, V, VI, and VII for sequence determination can be achieved without employing sodium dodecyl sulfate. The method presented essentially involves pyridine extraction, pH fractionation, ammonium sulfate fractionation, and various types of column chromatography. However, subunits I and II can be prepared only in the presence of sodium dodecyl sulfate by molecular sieve chromatography; subunit III can also be isolated in this manner. The separation of subunits is found to be hindered by phospholipids associated with the enzyme and therefore the phospholipid-depleted preparation is used as the starting material. The molecular weights of subunits I, II, III, IV, V, VI, and VII are 40,000, 21,000, 14,800, 13,500, 11,600, 9,500, and 7,600, respectively. These values are based on the results of the conventional Weber and Osborn method of gel electrophoresis in the presence of sodium dodecyl sulfate. The amino termini of subunits I and II have been determined as N-formylmethionine, and those of subunits III, IV, V, VI, and VII are alanine, alanine, serine, alanine, and an N acetyl-blocked residue, respectively. The carboxyl termini for subunits I to VII are lysine, leucine, lysine, histidine, valine, isoleucine, and valine, respectively. The complete amino acid sequence of some subunits has been published and that of other subunits will be reported elsewhere in collaboration with the Amino Acid Sequence Group of Cytochrome Oxidase at the University of Hawaii. PMID- 6270142 TI - In vitro transcription of the yeast alcohol dehydrogenase I gene by homologous RNA polymerase B (II). Selective initiation and discontinuous elongation on a supercoiled template. AB - A new in vitro approach is used to investigate the specificity of purified yeast RNA polymerase B (II). The template is supercoiled, the transcription is primed by a dinucleotide, and the transcripts are analyzed by polyacrylamide gel electrophoresis after synthesis in the absence of one nucleoside triphosphate. Under these conditions, two recombinant plasmids carrying the gene or part of the gene for yeast alcohol dehydrogenase I direct the synthesis of a very limited number of oligonucleotides. Elongation of these prelabeled oligomers, using unlabeled substrates, occurs in a discontinuous way. A major transcript of 200 nucleotides accumulates transiently. Southern hybridization shows that it is initiated about 1,400 bases upstream from the origin of the yeast alcohol dehydrogenase I gene. A minor start was identified, by a modified runoff experiment, at position -35 from the AUG initiation codon. The location of this site is related to the presumptive in vivo transcription starts. The selectivity disappears when the template is a truncated DNA. Then, initiation occurs predominantly at nicks introduced by the restriction enzymes. PMID- 6270143 TI - Dissociation kinetics of 125I-nerve growth factor from cell surface receptors. Acceleration by unlabeled ligand and its relationship to negative cooperativity. AB - The dissociation kinetics of 125I-labeled nerve growth factor (NGF) from dissociated cells of the 9-day chick embryo dorsal root ganglion were studied following preincubation for 2 h at 25 degrees C with 15 to 30 pM125I-NGF. Subsequent dilution in the presence of 4 nM NGF resulted in greater initial dissociation than occurred with dilution alone. This effect was largely limited to the first half-hour after addition of unlabeled NGF; thereafter, cell associated radioactivity declined at the same rate for both dilution conditions. In addition, dissociation kinetics did not follow a single first order rate law in the presence or absence of excess unlabeled NGF. These data are not consistent with a model of a single class of high affinity NGF receptors, whether or not they are noninteracting or negatively cooperative. Alternative models based on multiple noninteracting or negatively cooperative receptors might explain these results, but the current data do not allow clear discrimination among them. PMID- 6270144 TI - Membrane topology of beef-heart ubiquinone-cytochrome c reductase (complex III). AB - The membrane topology of ubiquinone-cytochrome c reductase (EC 1.10.2.2.) has been investigated with photoreactive lipid analogs (Bisson, R., and Montecucco, C. (1981) Biochem. J. 193, 757-763), both in its isolated form and when part of succinate-cytochrome c reductase (Complex II + III). These probes react specifically with those polypeptide chains exposed to lipids, thereby labeling them radioactively. Highly resolving gel electrophoretic conditions have been used to determine the patterns of labeling. Core protein I, cytochrome b, cytochrome c1, and polypeptides VI, VII, VIII, and IX contribute to the lipid protein boundary of Complex III. Evidence that the interaction between Complex II and Complex III involves their hydrophobic domains is also presented. PMID- 6270145 TI - Induction of delta-aminolevulinate synthase and cytochrome P-450 hemoproteins in hepatocyte culture. Effect of glucose and hormones. AB - Addition of glucose to cultured chick embryo hepatocytes caused a concentration dependent impairment of phenobarbital-mediated induction of delta-aminolevulinate (ALA) synthase resembling the "glucose effect" observed in rodents in vivo. This glucose effect occurred in the complete absence of extrahepatic factors such as serum and hormones. Fructose, glycerol, and lactate mimicked the inhibitory glucose effect on ALA synthase induction, whereas 2-deoxyglucose and 3-O methylglucose augmented the induction evoked by phenobarbital. 2-Deoxyglucose reversed the effect of glucose, glycerol, and lactate on ALA synthase induction suggesting that the glucose effect is mediated by free glucose or glucose 6 phosphate or a nonglycolytic metabolite of glucose 6-phosphate. The phenobarbital mediated induction of cytochrome P-450 hemoprotein(s) and its monooxygenase function were concomitantly diminished by glucose. However, this inhibitory effect or glucose was reversible by the addition of exogenous heme or ALA suggesting that the primary target of the glucose effect is ALA synthase induction and not synthesis of apocytochrome P-450. Glucagon and dibutyryl cAMP enhanced the induction of ALA synthase and cytochrome P-450 by phenobarbital and partially counteracted the glucose effect on both enzymes suggesting that the glucose effect may be mediated by changes in cAMP levels. Although insulin did not alter induction of ALA synthase, it impaired induction of cytochrome P-450 even in the presence of glucagon and cAMP. These data may be relevant for the treatment with glucose and heme of patients with "inducible" hepatic porphyria. PMID- 6270146 TI - Nucleotide sequence of rat alpha 1-acid glycoprotein messenger RNA. AB - The complete nucleotide sequence of rat alpha 1-acid glycoprotein (alpha 1-AGP) mRNA has been determined from cloned double-stranded cDNA. The coding portion of the mRNA was bounded at the ends by a 5'-untranslated region of 35 nucleotides in length and a 3'-untranslated region of 119 nucleotides in length. The 3' untranslated region contains the characteristic AAUAAA sequence ending 18 nucleotides from the 3'-terminal poly(A) segment. The 5'-region of the mRNA contains two in-phase AUG codons separated by 12 nucleotides. Comparison with the known NH2-terminal amino acid sequence of serum rat alpha 1-AGP suggests that the primary translation product of the mRNA contains an additional 14 or 18 amino acids that are not present in the mature form of the protein, which contains 187 amino acids. The inferred amino acid sequence of rat alpha 1-AGP and the known amino acid sequence of human alpha 1-AGP have several regions of identity clustered in the NH2-terminal portion of the proteins. The carboxyl-terminal regions show significantly less homology. Six potential asparagine glycosylation sites are found in the rat sequence, and four of these sites are in positions similar to known glycosylation sites in the human protein. Furthermore, three of these potential glycosylation sites are in a region that exhibits extensive amino acid sequence conservation, suggesting that this region may be important for the biological function of alpha 1-AGP. PMID- 6270147 TI - Accurate in vitro transcriptional initiation of the chick alpha 2 (Type I) collagen gene. AB - Chick genomic DNA containing the extreme 5' end of the alpha 2 (type I) collagen gene has been used as template in an in vitro HeLa cell transcription system. RNA polymerase II-dependent transcription initiates from a specific site on this DNA. The precise location of this site was determined by three types of experiments: sizing of in vitro-synthesized RNA runoff transcripts, comparing the sequence of the in vitro-made RNA transcripts with the structure of the DNA template, and identifying the first and second nucleotides of the in vitro-synthesized transcripts. Transcription was found to initiate 33 base pairs downstream from a canonical Goldberg-Hogness sequence (TATAAATA). This in vitro start site is the same as the initiation site of in vivo-synthesized collagen RNA. PMID- 6270148 TI - Plasmid-directed expression of Staphylococcus aureus beta-lactamase by Bacillus subtilis in vitro. AB - A plasmid carrying the Gram-positive Staphylococcus aureus PC1 beta-lactamase gene is active in directing a cell-free transcription and translation system from Bacillus subtilis. The major protein synthesized has been identified as the S. aureus beta-lactamase on the basis of peptide mapping. The protein is larger than the extracellular enzyme by about Mr = 3100. Significant in vitro translation of the beta-lactamase mRNA occurs in the absence of the initiation factor fraction as is characteristic of translation of mRNAs of Gram-positive origin. The 1250 base transcript that encodes the beta-lactamase and leader sequence has been mapped on the plasmid molecule. PMID- 6270149 TI - Calmodulin-activated cyclic nucleotide phosphodiesterase from brain. Changes in molecular size assessed by gel filtration and electrophoresis. PMID- 6270150 TI - Calmodulin-activated cyclic nucleotide phosphodiesterase from brain. Relationship of subunit structure to activity assessed by radiation inactivation. AB - The apparent target sizes of the basal and calmodulin-dependent activities of calmodulin-activated phosphodiesterase from bovine brain were estimated using target theory analysis of data from radiation inactivation experiments. Whether crude or highly purified samples were irradiated, the following results were obtained. Low doses of radiation caused a 10 to 15% increase in basal activity, which, with further irradiation, decayed with an apparent target size of approximately 60,000 daltons. Calmodulin-dependent activity decayed with an apparent target size of approximately 105,000 daltons. The percentage stimulation of enzyme activity by calmodulin decreased markedly as a function of radiation dosage. These observations are consistent with results predicted by computer assisted modeling based on the assumptions that: 1) the calmodulin-activated phosphodiesterase exists as a mixture of monomers which are fully active in the absence of calmodulin and dimers which are inactive in the absence of calmodulin; 2) in the presence of calmodulin, a dimer exhibits activity equal to that of two monomers; 3) on radiations destruction of a dimer, an active monomer is generated. This monomer-dimer hypothesis provides a plausible explanation for and definition of basal and calmodulin-dependent phosphodiesterase activity. PMID- 6270151 TI - Studies on the phosphorylation and synthesis of type I regulatory subunit of cyclic AMP-dependent protein kinase in intact S49 mouse lymphoma cells. AB - Phosphorylation and synthesis of type I regulatory subunit (RI) of cAMP-dependent protein kinase were studied using two-dimensional polyacrylamide gel electrophoresis of [35S]methionine-labeled proteins from intact S49 mouse lymphoma cells. [32P]Phosphate labeling, peptide mapping, and acid hydrolysis confirm that charge heterogeneity in RI results from phosphorylation of a single serine residue. In drug-free cells, phosphorylation proceeds to a steady state proportion of 90 to 95% of total RI with a half-time of about 25 min. The rate and steady state extent of RI phosphorylation are reduced by some, but not all, agents causing intracellular kinase activation. These results suggest that RI might assume different conformations in association with different amounts of cAMP or different analogs of cAMP. Endogenous kinase activation has no immediate effect on RI synthesis but leads to a moderate increase in RI synthesis after several hours; this induction occurs with all agents tested. Mutants of S49 cells lacking catalytic activity of cAMP-dependent protein kinase exhibit reduced phosphorylation and synthesis of RI. Comparative studies suggest that the phosphorylation of RI and its induction by kinase activation are fairly general phenomena; the extent of RI phosphorylation and the relative rate of RI synthesis are variable among cell types. PMID- 6270152 TI - Association of a 68,000-dalton protein with adrenocorticotropin-sensitive adenylate cyclase activity in Y1 adrenocortical tumor cells. AB - This report explores the biochemical basis for clonal variation in adrenocorticotropin (ACTH)-sensitive adenylate cyclase activity in the Y1 mouse adrenocortical tumor cell line. We demonstrate that the level of a specific protein, designated p68, is significantly correlated with the ability of adrenocorticotropin to stimulate adenylate cyclase activity among Y1 subclones (p = 0.004; r = 0.65). p68 was characterized by its molecular weight in sodium dodecyl sulfate polyacrylamide gels (Mr = 68,000) and by its isoelectric point as determined by two-dimensional gel electrophoresis (pI = 7.2). On two-dimensional gels, the protein migrated as a major spot with satellite spots 0.1 pH unit on either side. Homogenates and plasma membrane fractions from clones highly responsive to ACTH had large amounts of p68. In homogenates from highly responsive clones p68 represented 10 to 12% of the total protein. Homogenates and plasma membrane fractions from clones insensitive to ACTH were deficient in p68. In homogenates from the insensitive clones Y6 and OS3, p68 represented less than or equal 0.8% of the total protein. A somatic cell hybrid, formed by fusion of these two ACTH-insensitive clones recovered ACTH-sensitive adenylate cyclase activity and concomitantly expressed appreciable levels of p68. It is suggested that p68 may regulate the transfer of information from the occupied ACTH receptor ot the catalytic subunit of adenylate cyclase. PMID- 6270153 TI - Microfilament-mediated surface change in starfish oocytes in response to 1 methyladenine: implications for identifying the pathway and receptor sites for maturation-inducing hormones. AB - Oocytes of the starfish Pisaster ochraceus exhibit an early response to 1 methyladenine (the maturation-inducing hormone), which is described for the first time. In this response approximately 6,500 spikelike surface projections, much larger than microvilli, emerge transiently from oocytes stripped of their follicle cells and then treated with the hormone in vitro. Each spike contains a prominent bundle of microfilaments, possibly composed of actin. The distribution of spikes when follicle cells are only partially removed and the morphological details of the normal junctional association between follicle cells and oocytes suggest that 1-methyladenine-sensitive sites (receptor sites) can be identified with the approximately 6,500 postjunctional specializations that are part of the oocyte surface. This finding in turn is employed to construct a set of hypotheses concerning the route that 1-methyladenine normally takes from the follicle cells to an oocyte during stimulation of maturation; it is postulated that, for each oocyte, 1-methyladenine is transported along approximately 6,500 thin follicle cell processes, it is transmitted across the junctional gaps of an equivalent number of junctions between follicle cells and an oocyte, and then interacts with the postjunctional sites where 1-methyladenine receptors are thought to be clustered. Comparative aspects of this mode of intercellular communication are discussed. PMID- 6270154 TI - A role for calmodulin in the regulation of steroidogenesis. AB - TWO APPROACHES WERE USED TO STUDY THE POSSIBLE ROLE OF CALMODULIN IN THE REGULATION OF STEROID SYNTHESIS BY MOUSE ADRENAL TUMOR CELLS: trifluoperazine was used as an inhibitor of calmodulin and liposomes were used to deliver calmodulin into the cells. Trifluoperazine inhibits three steroidogenic responses to both ACTH and dibutyryl cyclic AMP: (a) increase in steroid production, (b) increased transport of cholesterol to mitochondria, and (c) increased side-chain cleavage by mitochondria isolated from cells incubated with ACTH or dibutyryl cyclic AMP. When calmodulin is introduced into the cells via liposomes, steroid synthesis is slightly stimulated. When calmodulin extensively dialyzed against EGTA, this stimulation is abolished. Ca(2+) introduced via liposomes was also without effect. However, when both calmodulin and Ca(2+) are introduced via liposomes (either in separate liposomes or in the same liposomes), steroid synthesis is stimulated. This stimulation does not occur when either anticalmodulin antibodies or EGTA is also present in the liposomes or when trifluoperazine is present in the incubation medium. Calmodulin and Ca(2+) presented together in liposomes to the cells stimulate transport of cholesterol to mitochondria, and side-chain cleavage activity is greater in mitochondria isolated from cells previously fused with liposomes containing calmodulin and Ca(2+) than in mitochondria from cells fused with liposomes containing buffer only. These observations suggest that calmodulin may be involved in regulating the transport of cholesterol to mitochondria, a process which is stimulated by ACTH and dibutyryl cyclic AMP and which may account, at least in part, for the increase in steroid synthesis produced by these agents. PMID- 6270155 TI - Microtubule-associated proteins (MAPs) and the organization of actin filaments in vitro. AB - When purified muscle actin was mixed with microtubule-associated proteins (MAPs) prepared from brain microtubules assembled in vitro, actin filaments were organized into discrete bundles, 26 nm in diameter. MAP-2 was the principal protein necessary for the formation of the bundles. Analysis of MAP-actin bundle formation by sedimentation and electrophoresis revealed the bundles to be composed of approximately 20% MAP-2 and 80% actin by weight. Transverse striations were observed to occur at 28-nm intervals along negatively stained MAP actin bundles, and short projections, approximately 12 nm long and spaced at 28 nm intervals, were resolved by high-resolution metal shadowing. The formation of MAP-actin bundles was inhibited by millimolar concentrations of ATP, AMP-PCP (beta, gamma-methylene-adenosine triphosphate), and pyrophosphate but not by AMP, ADP, or GTP. The addition of ATP to a solution containing MAP-actin bundles resulted in the dissociation of the bundles into individual actin filaments; discrete particles, presumably MAP-2, were periodically attached along the splayed filaments. These results demonstrate that MAPs can bind to actin filaments and can induce the reversible formation of actin filament bundles in vitro. PMID- 6270156 TI - A protein kinase bound to the projection portion of MAP 2 (microtubule-associated protein 2). AB - In previous work we have demonstrated that the microtubule-associated protein 2 (MAP 2) molecule consists of two structural parts. One part of the molecule, referred to as the assembly-promoting domain, binds to the microtubule surface and is responsible for promoting microtubule assembly; the other represents a filamentous projection observed on the microtubule surface that may be involved in the interaction of microtubules with other cellular structures. MAP 2 is known to be specifically phosphorylated as the result of a protein kinase activity that is present in microtubule preparations. We have now found that the activity copurifies with the projection portion of MAP 2 itself. Kinase activity coeluted with MAP 2 when microtubule protein was subjected to either gel- filtration chromatography on bio-gel A-15m or ion-exchange chromatography on DEAE- Sephadex. The activity was released from microtubules by mild digestion with chymotrypsin in parallel with the removal by the protease of the MAP 2 projections from the microtubule surface. The association of the activity with the projection was demonstrated directly by gel filtration chromatography of the projections on bio gel A-15m. Three protein species (M(r) = 39,000, 55,000, and 70,000) cofractionated with MAP 2, and two of these (M(r) = 39,000 and 55,000) may represent the subunits of an associated cyclic AMP- dependent protein kinase. The projection-associated activity was stimulated 10-fold by cyclic AMP and was inhibited more than 95 percent by the cyclic AMP-dependent protein kinase inhibitor from rabbit skeletal muscle. It appeared to represent the only significant activity associated with microtubules, almost no activity being found with tubulin, other MAPs, or the assembly-promoting domain of MAP 2, and was estimated to account for 7-22 percent of the total brain cytosolic protein kinase activity. The location of the kinase on the projection is consistent with a role in regulating the function of the projection, though other roles for the enzyme are also possible. PMID- 6270157 TI - Fluorescent low density lipoprotein for observation of dynamics of individual receptor complexes on cultured human fibroblasts. AB - The visible wavelength excited fluorophore 3,3'-dioctadecylindocarbocyanine iodide (Dil[3]) was incorporated into human low density lipoprotein (LDL) to form the highly fluorescent LDL derivative dil(3)-LDL. Dil(3)-LDL binds to normal human fibroblasts and to human fibroblasts defective in LDL receptor internalization but does not bind to LDL receptor-negative human fibroblasts at 4 degrees C or 37 degrees C. It is internalized rapidly at 37 degrees C by normal fibroblasts and depresses the activity of 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMG-CoA reductase) in a manner similar to that of LDL. It is prevented from binding to the LDL receptor by an excess of unlabeled LDL or by heparin sulfate. Identical distributions of dil(3)-LDL are observed on cells by either indirect immunofluorescence with fluorescein-labeled antibody or directly by dil(3) fluorescence. Upwards of 45 molecules of dil(3) are incorporated per molecule of LDL without affecting binding to the receptor. This labeling renders individual molecules visible by their fluorescence and enables the derivative to be used in dynamic studies of LDL-receptor motion on living fibroblasts by standard fluorescence techniques at low LDL receptor density. Observations with this derivative indicate that the LDL-receptor complex is immobilized on the surface of human fibroblasts but, when free of this linkage, undergoes a Brownian motion consistent with theory. PMID- 6270158 TI - Galactose-specific recognition system of mammalian liver: receptor distribution on the hepatocyte cell surface. AB - An isolated perfused liver system was used to study the distribution of asialoglycoprotein (ASGP) binding sites on rat hepatocyte cell surfaces. The number of surface receptors was quantitated by monitoring clearance of 125I labeled ligands from the perfusate medium under two conditions that blocked their internalization: low temperature (less than 5 degrees C) or brief formaldehyde fixation. The cell surface distribution of binding sites was visualized in the electron microscope with either asialoorosomucoid covalently coupled to horseradish peroxidase (ASOR-HRP) or lactosaminated ferritin (Lac-Fer), both of which were bound with similar kinetics and to similar extents as ASOR itself. At low temperature or after prefixation, ASGP binding sites were present over much of the sinusoidal cell surface, but were concentrated most heavily over coated pits. Quantitation of ligand distribution at 4 degrees C with Lac-Fer gave an approximately 70-fold greater density of ferritin particles over coated membrane than over uncoated regions. We obtained no evidence for gradual movement of ASGP receptors into or out of coated pits within the time-course of our experiments. Finally, the number and distribution of cell surface binding sites was unaffected by previous exposure to ASOR or by inhibition of endocytic vesicle-lysosome fusion and ASOR degradation at 16 degrees C. PMID- 6270159 TI - Transient activity of Golgi-like membranes as donors of vesicular stomatitis viral glycoprotein in vitro. AB - Previous reports demonstrated that the vesicular stomatitis viral glycoprotein (G protein), initially present in membranes of a Chinese hamster ovary mutant cell line (clone 15B) that is incapable of terminal glycosylation, can be transferred in vitro to exogenous Golgi membranes and there glycosylated (E. Fries and J. E. Rothman, 1980, Proc. Natl. Acad. Sci. U. S. A. 77:3870-3874; and J. E. Rothman and E. Fries, 1981, J. Cell Biol. 89:162-168). Here we present evidence that Golgi-like membranes serve as donors of G protein in this process. Pulse-chase experiments revealed that the donor activity of membranes is greatest at approximately 10 min of chase, a time when G protein has been shown to have arrived in Golgi stacks (J. E. Bergmann, K. T. Tokuyasu, and S. J. Singer, 1981, Proc. Natl. Acad. Sci. U. S. A. 78:1746-1750). Additional evidence that the G protein that is transferred to exogenous Golgi membranes in vitro had already entered the Golgi membranes in vivo was provided by observations that its oligosaccharides had already been trimmed, and that its distribution in a sucrose density gradient was coincident with that of enzymatic markers of Golgi membranes. The capacity of this Golgi-like membrane to serve as donor is transient, declining within 5 min after "trimming" in vivo as the G protein enters a "nontransferable" pool. The rapidity of the process suggests that both the "transferable" and "nontransferable" pools of G protein reside in Golgi-like membranes. PMID- 6270160 TI - Preferential phosphorylation of the 150,000 molecular weight component of neurofilaments by a cyclic AMP-dependent, microtubule-associated protein kinase. AB - Highly purified preparations of bovine brain and rabbit nerve root neurofilaments were found to be lacking in protein kinase activity when either histone FIIA or the neurofilaments themselves were used as acceptors. There was no augmentation of activity in the presence of cyclic AMP. Addition of microtubule proteins prepared by cycles of assembly and disassembly resulted in phosphorylation of histone, phosphorylation of tubulin and the microtubule-associated proteins, and phosphorylation of neurofilament subunits. The phosphorylation of neurofilaments was predominantly in the 150,000-dalton species and was completely cyclic AMP dependent. PMID- 6270161 TI - Control of cell volume in the J774 macrophage by microtubule disassembly and cyclic AMP. AB - We have explored the possibilities that cell volume is regulated by the status of microtubule assembly and cyclic AMP metabolism and may be coordinated with shape change. Treatment of J774.2 mouse macrophages with colchicine caused rapid microtubule disassembly and was associated with a striking increase (from 15-20 to more than 90 percent) in the proportion of cells with a large protuberance at one pole. This provided a simple experimental system in which shape changes occurred in virtually an entire cell population in suspension. Parallel changes in cell volume could then be quantified by isotope dilution techniques. We found that the shape change caused by colchicine was accompanied by a decrease in cell volume of approximately 20 percent. Nocodozole, but not lumicolchicine, caused identical changes in both cell shape and cell volume. The volume loss was not due to cell lysis nor to inhibition of pinocytosis. The mechanism of volume loss was also examined. Colchicine induced a small but reproducible increase in activity of the ouabain-sensitive Na(+), K(+)-dependent ATPase. However, inhibition of this enzyme/transport system by ouabain did not change cell volume nor did it block the colchicines-induced decrease in volume. One the other hand, SITS (4'acetamido, 4-isothiocyano 2,2' disulfonic acid stilbene), an inhibitor of anion transport, inhibited the effects of colchicines, thus suggesting a role for an anion transport system in cell volume regulation. Because colchicine is known to activate adenylate cyclase in several systems and because cell shape changes are often induced by hormones that elevate cyclic AMP, we also examined the effects of cyclic AMP on cell volume. Agents that act to increase syclic AMP (cholera toxin, which activates adenylate cyclase; IBMX, and inhibitor of phosphodiesterase; and dibutyryl cyclic AMP) all caused a volume decrease comparable to that of colchicine. To define the effective metabolic pathway, we studied two mutants of J774.2, one deficient in adenylate cyclase and the other exhibiting markedly reduced activity of cyclic AMP-dependent protein kinase. Cholera toxin did not produce a volume change in either mutant. Cyclic AMP produced a decrease in the cyclase-deficient line comparable to that in wild type, but did not cause a volume change in the kinase- deficient line. This analysis established separate roles for cyclic AMP and colchicine. The volume decrease induced by cyclic AMP requires the action of a cyclic AMP-dependent protein kinase. Colchicine, on the other hand, induced a comparable volume change in both mutants and wild type, and thus does not require the kinase. PMID- 6270162 TI - Hepatic binding and internalization of low density lipoprotein-gold conjugates in rats treated with 17 alpha-ethinylestradiol. AB - Receptor-mediated hepatic uptake of low density lipoproteins (LDL) conjugated to colloidal gold was studied by perfusion of livers from rats treated for 5 d with 17 alpha-ethinylestradiol. Estrogen treatment resulted in a marked decrease in serum lipid and lipoprotein concentrations. After 15 min of perfusion the conjugate was bound to the hepatic microvilli of both control and estrogen treated rats; the estrogen-treated rats showed an 8- to 11-fold greater number of membrane-bound conjugates. The conjugates were bound to the membrane receptor by the LDL particle because the gold granules were regularly displaced from the membrane by 20 +/- 3.2 nm, the diameter of LDL. Internalization of the conjugate, evident by gold particles in multivesicular bodies, occurred at coated pits at the base of the microvillus where coated vesicles containing a single gold-LDL conjugate were released. After 1 h of perfusion, the livers from the estrogen treated rats showed all phases of endocytosis and incorporation into multivesicular bodies of the conjugate. After 2 h of perfusion, there was congregation of gold-labeled lysosomes near the bile canaliculi. Gold-LDL conjugates were also observed to bind and be internalized by Kupffer cells and sinusoidal endothelium. These findings indicate that estrogen treatment induces hepatic receptors for LDL. The catabolic pathway of binding and endocytosis of the conjugate is similar to that seen in fibroblasts, although slower. Because gold-LDL conjugates were also present in the Kupffer and endothelial cells, the uptake of LDL by the liver involves the participation of more than a single cell type. PMID- 6270163 TI - F-actin aggregates in transformed cells. AB - Polymerized actin has been found aggregated into distinctive patches inside transformed cells in culture. The F-actin-specific fluorescent probe, nitrobenzoxadiazole-phallacidin, labels these F-actin aggregates near the ventral cell surface of cells transformed by RNA or DNA tumor viruses, or by chemical mutagens, or spontaneously. Their appearance in all eight transformed cell types studied suggests their ubiquity and involvement in transformation morphology. Actin patches developed in normal rat kidney (NRK) cells transformed by a temperature-sensitive mutant of Rous sarcoma virus (LA23-NRK) within 30 min after a shift from the nonpermissive (39 degrees C) to the permissive temperature (32 degrees C). Patch appearance paralleling viral src gene expression tends to implicate pp60src kinase activity in destabilizing the cytoskeleton. However, appearance of the actin aggregates in cells not transformed by retrovirus calls for alternative mechanisms, perhaps involving an endogenous kinase, for this apparently common trait. PMID- 6270164 TI - Expression of angiotensin-converting enzyme activity in cultured pulmonary artery endothelial cells. AB - Angiotensin-converting enzyme (EC 3.4.51.1) is a carboxyterminal dipeptidyl peptidase. The enzyme catalyzes the conversion of the decapeptide angiotensin I to the octapeptide angiotensin II. In addition, the enzyme catabolizes bradykinin. Because of these actions, the enzyme is of pivotal importance in blood pressure homeostasis. Numerous investigators have demonstrated the presence of the enzyme in association with endothelial cells but relatively little is known concerning the factors controlling the expression enzyme activity by endothelial cells in culture. We have demonstrated that endothelial cells in culture do not express significant amounts of enzyme activity until several days after growth ceases due to high cell density. This is important because it demonstrates a change in function with stage of growth in culture and a possible difference in functional capabilities between nondividing endothelial cells and cells that are dividing in response to injury. Since density-dependent expression of differentiated traits does not appear to be unique to endothelial cells an understanding of the mechanisms underlying this phenomenon may provide a general explanation for the expression of differentiated traits by cultured cells. PMID- 6270165 TI - DNA synthesis in primary cultures of adult rat hepatocytes in a defined medium: effects of epidermal growth factor, insulin, glucagon, and cyclic-AMP. AB - Epidermal growth factor (EGF) especially in combination with insulin and glucagon, has been shown to stimulate DNA synthesis in liver cells, both in the whole animal and in cell cultures. As a further development we have found that in primary monolayer cultures of freshly isolated adult rat liver parenchymal cells, in which contamination with nonparenchymal cells was negligible, DNA synthesis was substantially stimulated by these substances. In control cultures, incorporation of [3H]thymidine into DNA and labeling of nuclei in autoradiographs was low. The stimulation by EGF was enhanced by insulin and glucagon, whereas these hormones by themselves exhibited only limited activity. These observations were made in cultures of hepatocytes that were never exposed to serum, even during cell isolation and plating. Hence for stimulation of DNA synthesis under these conditions neither serum factors nor interactions with other types of cells or their products were required. The effects of glucagon were reproduced by substances that elevate intracellular concentration of cyclic-AMP, including cholera toxin, isoproterenol, and methylisobutylxanthine. These various substances, especially EGF, glucagon, or cyclic-AMP, altered the morphological characteristics of the cultures during early stages, promoting cellular spreading and aggregation. PMID- 6270166 TI - Anchorage-independent growth of normal calf lens epithelial cells and effect of SV40 transformation on their growth properties. AB - Calf lens epithelial cells cultured in vitro show growth properties usually associated with virally transformed fibroblasts. The lens cells are anchorage independent, forming colonies in agar, and show a low requirement for added mitogens. In dense culture they form multilayers and maintain a constant cell number by proliferation and shedding. Strains of lens cells transformed by SV40 virus have been obtained that show similar growth properties to the normal lens epithelium. The major effect of SV40 transformation is to increase the growth rate, final cell density and in vitro life-span of the lens cells and to inhibit the increase in size that occurs after 2-3 weeks of culturing the untransformed cells. PMID- 6270167 TI - Differences in oxygen-dependent regulation of enzymes between tumor and normal cell systems in culture. AB - Metabolic studies in tumor cells have indicated that bioenergetic regulatory mechanisms geared to acute changes in oxygen availability are abnormal. In the present studies we have examined bioenergetic adaptations to chronic oxygen depletion in culture maintained tumor cells in comparison to normal cell lines. Activities of two key glycolytic enzymes (pyruvate kinase (PyKI) and phosphofructokinase (PFK)) were measured in two tumor cell lines (fibrosarcoma (FS) and Hela) and two normal cell lines (rat lung fibroblasts (RLF) and WI-38) maintained in culture for up to 96 hours under aerobic (PO2 approximately 140) and hypoxic PO2 approximately 15) conditions. Exposure to low O2 tensions for 96 hours resulted in significant increases in PyKi and PFK in both RLF and WI-38, ut did not alter activities of these enzymes in either FS or HeLa cell systems. Activities of two enzymes involved in O2 metabolism (cytochrome oxidase (CyOx) and superoxide dismutase (SOD) were also measured in the two tumor cell lines and in RLF. chronic hypoxia significantly decreased the activities of CyOx and SOD in RLF cell systems but did not alter the activities of these enzymes in the tumor cells. In these studies, the tumor-derived cell lines do not demonstrate specific enzymatic responses to sustained oxygen depletion in vitro noted in normal cell systems, suggesting significant abnormalities in regulatory mechanisms geared to chronic changes in molecular O2. PMID- 6270168 TI - Cellular regulatory role of leukotriene B4: its effects on cation homeostasis in rabbit neutrophils. AB - We have found that exogenous leukotriene B4 modifies calcium homeostasis in rabbit neutrophils in a manner essentially analogous to that of the chemotactic peptide f-Met-Leu-Phe. Leukotriene B4 causes a rapid and dose-dependent increase in membrane permeability to calcium and a release of calcium from previously unexchangeable intracellular pool(s). The net result of these changes is to transiently elevate the intracellular level of exchangeable calcium. A stereoisomer of leukotriene B4 with greatly reduced secretory activity toward neutrophils (5S, 12S-di HETE) is essentially without effect on the rate of 45Ca uptake at concentrations equal to those that produce near maximal enhancement by leukotriene B4. Leukotriene B4, in addition to its effects on calcium metabolism, also increases the rate of 22Na influx into rabbit neutrophils. The relationships between the action of leukotriene B4 on calcium homeostasis and the neutrophil directed activities of arachidonic acid and its lipoxygenase metabolites are discussed PMID- 6270169 TI - Mechanism of synergistic induction of DNA synthesis by epidermal growth factor and tumor promoters. AB - Evidence is presented to support our previously proposed hypothesis that the hyperplastic effect of tumor promoters is related to their ability to alter existing physiological levels of growth factors in target tissues. Epidermal growth factor and phorbol ester tumor promoters acted synergistically at low (0.001-0.05 ng/ml) but not high (greater than 0.1 ng/ml) EGF concentrations to induce DNA synthesis in cultured Rat-1 fibroblast cells. The degree of synergism correlated with the tumor-promoting ability of the compound. The tumor promoters decreased 125I-EGF binding to cellular receptors in a dose-dependent manner that also correlated with the tumor-promoting ability of the compound. The inhibition of EGF binding by phorbol ester compounds resulted in a decrease in the amount of EGF degraded as compared to control cultures. At limiting EGF concentrations, the sparing of EGF degradation resulted in an increase in the amount of EGF remaining in the culture medium after 12 h of incubation and a concomitant increase in the amount of EGF bound to phorbol ester-treated cells at this time as compared to control cultures. The ability of a phorbol ester compound to alter EGF degradation and to stimulate DNA synthesis synergistically with EGF correlated with the tumor-promoting ability of the compound and occurred only a low EGF concentrations. PMID- 6270170 TI - Acromegaly and Cushing's syndrome associated with a foregut carcinoid tumor. AB - We report an 18-yr-old youth with a metastatic foregut carcinoid tumor, Cushing's syndrome, and hypersomatotropic gigantism. Administration of cyproheptadine caused a dramatic fall in urinary cortisol excretion and plasma ACTH levels associated with clinical remission of the Cushing's syndrome. GH secretion was not affected by cyproheptadine administration. Ectopic ACTH secretion was confirmed by RIA of tumor extracts and immunohistochemical demonstration of ACTH containing cells in hepatic metastases. There were two sources of GH production demonstrated in this patient. Ectopic secretion of GH by the carcinoid hepatic metastases was documented by both RIA and immunohistochemical techniques. A somatotrophic pituitary tumor was also present. The histological characteristics of this tumor suggest adenomatous hyperplasia rather than de novo neoplastic change as the likely mechanism of its pathogenesis. GH releasing factor-like activity was demonstrated in extracts of plasma and in extracts of the carcinoid tumor. We conclude that cyproheptadine exerted an effect on the ectopic ACTH producing cells but not on the ectopic GH-producing cells or on adenohypophyseal GH secretion. Production of a GH releasing factor-like activity by the carcinoid tumor may have caused the pituitary somatotrophic tumor. PMID- 6270171 TI - The control of steroidogenesis by human fetal adrenal cells in tissue culture. III. The effects of various hormonal peptides. AB - The effects upon production of cortisol and dehydroepiandrosterone (DHA) by human fetal adrenal cells in tissue culture were studied using commercial hCG (0.5 and 5 IU/ml), purified hCG (0.7-6.7 IU/ml), the alpha-subunit of hCG (200 and 1000 ng/ml), human GH (50 and 200 ng/ml), human PRL (0.1-100 ng/ml), alpha-MSH (0.1-10 ng/ml), corticotropin-like intermediate lobe peptide (200 ng/ml), human beta lipotropin (0.1 and 0.2 ng/ml), and beta-endorphin (100 ng/ml). Although each peptide was added to the culture medium in a concentration either similar to that observed in the fetal circulation or (where such information was not available) in amounts several times greater than those effective for ACTH in this system, none demonstrated any significant stimulation of steroid production. In particular, repeated studies with hCG showed that this hormone had no stimulating effect upon DHA production, neither in cultures of whole adrenals nor in cultures of separated fetal zone and definitive zone cells. Furthermore, none of these peptides showed a synergistic effect upon DHA production when they were added to cultures together with concentrations of alpha-ACTH-(1-24) (10(2)-10(3) pg/ml) previously demonstrated to represent the middle of the dose-response curve. Indeed, the only significant interactions with alpha-ACTH-(1-24) observed in these studies were a slight reduction in cortisol production produced by corticotropin-like intermediate lobe peptide and apparent inhibition of DHA production by beta-lipotropin and GH. The data do not lend credence to the suggestion that any of these peptides plays an important role in vivo in stimulating fetal adrenal steroidogenesis. PMID- 6270172 TI - Epinephrine deficiency in hypocorticotropic hypopituitary children. AB - In rats, adrenal medullary synthesis of epinephrine is impaired by ACTH deficiency and is not improved by replacement doses of glucocorticoid. We have evaluated plasma epinephrine and norepinephrine concentrations in 43 children, 8 15 yr old. These children were divided into 5 groups, with 6-10 per group: normals; children with isolated GH deficiency; hypopituitary children deficient in both GH and TSH; hypopituitary children deficient in GH, TSH, and ACTH; and short children without known organic disease. The deficiencies of ACTH and TSH were being treated with replacement doses of cortisol and T4. Plasma catecholamines were measured in the supine position at rest every other hour from 0800-1600 h, and after exercise in the standing position at 1000 h. Plasma norepinephrine levels, both at rest and after exercise, were normal in all four groups of short children. Resting and postexercise plasma epinephrine levels were reduced to 10-20% of normal in the hypocorticotropic hypopituitary patients, and were normal in the other three groups of short children. PMID- 6270173 TI - Effects of catechol estrogen infusions upon gonadotropin and prolactin concentrations in men. AB - To study the effects of catechol estrogens upon gonadotropin secretion, 2 hydroxyestrone (2-OHE1) and 2-hydroxyestradiol (2-OHE2) were administered iv to young adult men in a range of doses for 4 days. Blood samples were obtained for plasma LH, FSH, and PRL at 20-min intervals for 6 h before and at the end of the infusion period. 2-OHE1 had no effect upon gonadotropins or PRL in doses up to 1.6 mg/day; at 3.2 and 6.6 mg/day, it produced a slight suppression of LH and FSH, with no change in PRL. 2-OHE2 was generally ineffective at 100 micrograms/day, but doses from 200-800 micrograms/day suppressed gonadotropins, without changes in PRL. These infusions elevated 2-OHE1 and 2-OHE2 plasma levels to values comparable to those measured in late pregnancy. There were no associated effects upon blood pressure and only minimal changes in urinary catecholamine excretion. No effects that could be interpreted as antiestrogenic were observed. These results are consistent with the hypothesis that circulating catechol estrogens behave as weak estrogens in men. PMID- 6270174 TI - Control of proliferation of human fetal adrenal cells in vitro. AB - The influence of fibroblast growth factor (FGF) and epidermal growth factor (EGF) on the proliferation of cultured human fetal adrenal cells has been examined. Separated human definitive zone and fetal zone adrenal cells plated at low density in the presence of 10% serum and maintained on plastic culture dishes proliferated slowly. If the cultures were exposed to either FGF or EGF, the growth rate of the cells from each zone increased significantly. Half-maximal stimulation of cell proliferation for both zones occurred at a concentration of 3 X 10(-11) M for EGF and 8 X 10(-9) M for FGF. In addition, 125I-labeled EGF binding to both definitive and fetal zone cells demonstrated high affinity (Kd = 10(-9) M). To investigate the influence of an extracellular matrix (ECM) on cell proliferation, separated fetal adrenal cells maintained on plastic culture dishes were compared with cells maintained on a recently described ECM prepared from bovine corneal endothelial cells. Fetal adrenal cells maintained on the ECM had a significantly higher growth rate than cells maintained on plastic alone. These results demonstrate 1) the mitogenic role of EGF and FGF for human fetal adrenal cells, and 2) that the type of substrate upon which fetal adrenal cells are maintained has a profound influence on their proliferation. PMID- 6270175 TI - Impaired clearance of beta-lipotropin in uremia. AB - Immunoreactive ACTH and beta-lipotropin (beta-LPH) plasma concentrations are elevated in clinically stable chronic renal failure patients on hemodialysis (LPH: patients, 271.8 +/- 35.7 fmol ml-1; normal subjects; 6.6 +/- 0.5; ACTH: patients, 56.4 +/- 15.3; normal subjects, 19.4 +/- 1.7). To begin to study the etiology of such elevated levels, the MCR, apparent volume of distribution, and fractional rate of disappearance of synthetic human ACTH and highly purified human beta-LPH were determined in two clinically stable chronic renal failure patients on hemodialysis, after bolus simultaneous injection of both peptides. Biphasic disappearance curves were obtained for beta-LPH; triphasic for ACTH. The MCR of ACTH was within the range seen in normal subjects, whereas the MCR of beta LPH was less than one half the normal rate. The data indicate that a decrease in MCR (rather than an increase in pituitary secretory rate) may account for the higher plasma levels of beta-LPH in uremic patients. PMID- 6270176 TI - Hyperreninemic hypoaldosteronism in the critically ill: a new entity. AB - To define the changes in adrenal gland function during critical illness, we evaluated 28 severely ill patients with persistent hypotension who were hospitalized in a medical intensive care unit. The patients had increased plasma cortisol (mean +/- SE, 40.1 +/- 10.1 micrograms/dl). PRA was increased in all subjects (21.6 +/- 7.2 ng/ml.h); however, the plasma aldosterone concentration was inappropriately low in 18 of the subjects, with values ranging from 1-9 ng/dl, despite normal serum potassium concentrations (4.3 +/- 0.1 meq/liter) and increased concentrations of the aldosterone percursor, 18-hydroxycorticosterone. These 18 patients had hypotension associated with major infections and a high mortality rate (78%). Infusions of ACTH or angiotensin II were associated with a normal aldosterone response in only 2 of the 14 patients tested, also suggesting that the defect was probably at the level of the zone glomerulosa cell. Although infection was a common underlying illness, no other factors, such as dopamine administration, decreased angiotensin-converting enzyme activity, or increased aldosterone clearance, could be implicated as the cause of the phenomena. Thus, selective hypoaldosteronism in the presence of high renin levels exists in a substantial percentage of hypotensive critically ill patients. PMID- 6270177 TI - Recovery of hypothalamic-pituitary-adrenal function after long term suppression by aminoglutethimide and hydrocortisone. AB - Little data are available concerning recovery of adrenal function after prolonged inhibition of steroidogenesis by enzyme inhibitors. Aminoglutethimide (AG), a potent blocker of adrenal steroid biosynthesis, combined with physiological replacement doses of hydrocortisone (HC) is currently being used to treat women with metastatic breast carcinoma. We studied the time-course of recovery of hypothalamic-pituitary-adrenal function after prolonged drug therapy in 10 women. Fifteen hours after stopping AG-HC therapy, 0900 h serum cortisol levels were normal [12.9 +/- 3.4 (SEM) micrograms/100 ml], but increments observed after provocative stimulation were blunted. However, the cortisol responses to both insulin-induced hypoglycemia and cortrosyn stimulation normalized 36 and 42 h after stopping AG and HC therapy. In addition, the concentration of plasma ACTH peaked at 175 +/- 9.3 pg/ml 15 min after the nadir of hypoglycemia. Adrenal histology in two patients who died while on chronic AG and HC therapy showed hypertrophic cells with large amounts of finely vacuolated cytoplasm in the zona fasciculata but no other abnormalities. We conclude that recovery of the hypothalamic-pituitary-adrenal axis is complete within 36 h after discontinuing chronic AG and HC therapy. This is in contrast to the prolonged suppression observed after chronic therapy with pharmacological doses of glucocorticoids. PMID- 6270178 TI - Corticotropin/beta-lipotropin biosynthesis, processing, and release in Nelson's syndrome. AB - Biosynthesis and processing of ACTH/beta-lipotropin was studied in Nelson's syndrome pituitary tumor tissue grown in monolayer culture. Radiolabeled peptides were immunoprecipitated and fractionated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS/PAGE). Important findings include: 1) a virtual absence of 13K ACTH or 3.5K beta-endorphin production; 2) evidence indicating the presence of a 24-26K ACTH and beta-LPH containing intermediate (which implies a different order of processing from that reported in the mouse); 3) An extremely rapid rate of turnover and release of ACTH and beta-lipotropin (beta-LPH) similar to that of the mouse AtT20/D16v pituitary tumors. The latter finding is consistent with an intrinsic pituitary cell defect in the pathogenesis of this disorder. PMID- 6270179 TI - Characterization of a small molecular size urinary immunoreactive human chorionic gonadotropin (hCG)-like substance produced by normal placenta and by hCG secreting neoplasms. AB - Urine obtained from normal pregnant women as well as from patients with hCG secreting tumors frequently contains native hCG and free hCG subunits when separated on Sephadex G-100. In addition, a small amount of an immunoreactive, hCG-like, low molecular weight substance is usually observed in those chromatograms and represents less than 1% of the total immunoreactive hCG present. Two patients with widely metastatic hCG-secreting tumors excreted disproportionately large quantities of that low molecular weight substance, and that observation raised the possibility that this substance was a secretory and not a degradative product of the hCG molecule. The small immunoreactive hCG-like substance was subsequently characterized immunologically, biologically, and physically. The hCG fragment displayed a biphasic dose-response line in a homologous hCG RIA. The slope of the upper portion of the dose-response line was equal to that for native hCG, but the slope of the lower component of the dose response line was significantly different from that for hCG. The immunoreactive hCG substance cross-reacted with hCG beta but not with either hCG alpha or hCG beta carboxyl-terminus. The small molecular size immunoreactive hCG-like substance bound to Concanavalin A-Sepharose 4B and eluted with 0.2 M alpha-D methyl glucopyranoside, contained no significant intrinsic biological activity when tested in the in vitro Leydig cell bioassay and also failed to compete with labeled hCG for specific ovarian LH/hCG receptors. Consequently, that small urinary immunoreactive hCG substance behaved neither as a hCG agonist or antagonist. It exhibited a plasma half-life of 4.5 min when the appropriate Sephadex G-100 fractions were injected into immature female rats. The small molecular size immunoreactive hCG-like substance may be a secretory or breakdown product of hCG-secreting cells. PMID- 6270180 TI - Calmodulin in dispersed human parathyroid cells. AB - We investigated the presence and several of the properties of calmodulin in human parathyroid cells. Boiled extracts of such cell preparations contained a heat stable factor causing a 2- to 3-fold calcium-dependent stimulation of calmodulin deficient phosphodiesterase activity, which was parallel to that due to pure porcine calmodulin. This activation could be totally blocked by 10(-4) M trifluoperazine, with half-maximal inhibition at 3 X 10(-5) M, similar to the effects of this phenothiazine on porcine calmodulin. These results suggested the presence of calmodulin in human parathyroid cells. By comparison with known quantities of porcine calmodulin, human parathyroid cells contained 9-208 ng calmodulin/10(6) cells. The content of calmodulin in 3 normal parathyroid glands [65 +/- 8 (+/- SEM) ng/10(6) cells] did not differ significantly from that of 12 adenomas (61 +/- 16 ng/10(6) cells). Cells from 7 glands showing secondary hyperplasia, however, had significantly greater levels of calmodulin (164 +/- 11 ng/10(6) cells) than either normal cells or adenomas (P less than 0.001 and P less than 0.005, respectively). Extracts of human parathyroid cells caused half maximal stimulation of phosphodiesterase activity at 1.1-4.8 microM free calcium. The concentrations of calcium half-maximally activating phosphodiesterase (Ka) did not differ significantly for normal or abnormal cells (3.3 +/- 0.03 vs. 2.6 +/- 0.33; P greater than 0.3). Moreover, in 2 cases in which normal parathyroid tissue was obtained from patients with adenomas, the Ka values for calcium for the normal and abnormal cells were similar (3.3 vs. 2.5 and 3.4 vs. 2.5 microM, respectively). Finally, there was no significant correlation between either the content of calmodulin or the Ka for calcium and the set-point for calcium [the calcium concentration causing half-maximal inhibition of parathyroid hormone (PTH) release] or the maximal rate of PTH secretion for dispersed parathyroid cells. These results suggest that human parathyroid cells contain calmodulin, but provide no evidence for a role of this protein in the abnormal calcium-regulated PTH release in hyperparathyroidism. PMID- 6270181 TI - Effect of age on serum immunoreactive parathyroid hormone and its biological effects. AB - Immunoreactive parathyroid hormone (iPTH) levels, nephrogenous cAMP (ncAMP), and tubular maximum phosphate reabsorption (TmP) were measured in 10 young and 12 healthy volunteers. The fasting urinary calcium to creatinine ratio (Ca:Cr) was also quantitated as an index of bone resorption. Aging was attended by increased iPTH levels (6.9 +/- 0.8 vs. 3.4 +/- 0.4 mu leq/ml; P less than 0.01) as well as increased ncAMP levels (2.48 +/- 0.28 vs. 1.12 +/- 0.21 nmol/100 ml glomerular filtrate; P less than 0.005) and decreased TmP (2.9 +/- 0.2 vs. 4.1 +/- 0.2 mg/100 ml glomerular filtrate; P less than 0.005), indicating that the increased iPTH levels reflected the biological effects of the hormone. A significant positive correlation of iPTH and ncAMP and a significant negative correlation of iPTH and TmP were observed. The Ca:Cr was increased in the older volunteers (0.10 +/- 0.02 vs. 0.05 +/- 0.01; P less than 0.05). The elderly subjects had significantly decreased daily calcium ingestion, serum phosphate and albumin, and creatinine clearance. Our findings suggest that the increased biological effects of PTH in the elderly subjects may contribute to the increases in Ca:Cr and bone loss that occur with age. PMID- 6270182 TI - A met-enkephalin analog inhibits adrenocorticotropin secretion by cultured pituitary cells from a patient with Nelson's syndrome. AB - ACTH excretion by cultured nonenzymatically dispersed pituitary tumor cells from a patient with Nelson's syndrome was studied. Hormone release was suppressed by 74 +/- 6% by the addition of 1 microM of the met-enkephalin analog FK 33824, while naloxone (1 microM) stimulated ACTH release by 70 +/- 5%. Somatostatin, dexamethasone, bromocriptine, and cyproheptadine in a concentration of 1 microM each inhibited ACTH release by 25 +/- 2%, 35 +/- 2%, 52 +/- 2%, and 61 +/- 4%, respectively, while lysine vasopressin (0.1 microM) and dibutyryl cAMP (5 mM) stimulated ACTH release by 112 +/- 8% and 220 +/- 4%, respectively. In conclusion, it was shown that the stimuli mentioned above directly affect ACTH secretion by the pituitary tumor cells. The inhibitory action of the met enkephalin analog and the stimulatory action of naloxone on ACTH secretion make the presence of opiate receptors on this type of tumor likely. PMID- 6270183 TI - Effect of dibutyryl adenosine 3',5'-monophosphate administration on plasma concentrations of 1,25-dihydroxyvitamin D in pseudohypoparathyroidism type I. AB - The effect of dibutyryl cAMP on plasma concentrations of 1,25-dihydroxyvitamin D [1,25-(OH)2D] was studied in two patients with pseudohypoparathyroidism (PHP) type I, five normal adults, and three normal children as controls. In normal adults, plasma 1,25-(OH)2D tended to increase 3 h after the infusion of 2.5 mg/kg dbcAMP and was significantly increased after 6 h. In normal children, plasma 1,25 (OH)2D increased 3 h after the infusion, then gradually decreased; in the patients with PHP type I, it increased greatly from 6.8 and 11.2 pg/ml to 264.6 and 128.2 pg/ml 3 h after the infusion. These results suggest that the renal mechanism for the response to parathyroid hormone is intact distal to the renal adenylate cyclase in patients with PHP type I. PMID- 6270184 TI - Malignancy-associated hypercalcemia: evaluation with a cytochemical bioassay for parathyroid hormone. AB - Employing a cytochemical assay initially developed for measuring parathyroid hormone (PTH), bioactivity was assessed in 33 patients with malignancies. Initial studies in vitro were consistent with a role for cAMP as a second messenger in the bioassay. Cytochemical bioactivity was increased in the peripheral plasma of 10 of 16 hypercalcemic patients with elevated nephrogenous cAMP excretion, and mean levels were 10-fold higher in these patients than in 17 normocalcemic or hypercalcemic patients with normal or suppressed nephrogenous cAmP excretion, respectively. Plasma bioactivity, serum calcium, and nephrogenous cAMP excretion all fell to normal in 1 patient after tumor resection, and cytochemical bioactivity was demonstrable in the tissue culture medium in which the neoplasm was maintained. Gel chromatographic analysis revealed that a major component of plasma bioactivity eluted before rather than with PTH-(1-84) in patients with malignancy in contrast with that in patients with primary hyperparathyroidism. The studies, therefore, demonstrate the capacity of the cytochemical bioassay to measure increased activity in patients with malignancy, hypercalcemia, and elevated nephrogenous cAMP excretion; suggest that the material responsible for the activity differs from PTH-(1-84); and provide a sensitive detector system for further analysis of this material and its role in the pathogenesis of this disease. PMID- 6270185 TI - Presence of ectopic beta-adrenergic receptors on human adrenocortical cortisol producing adenomas. AB - A direct binding study of radioligand [3H]dihydroalprenolol (DHA), a potent beta adrenergic antagonist, was performed on the particulate fractions of four adrenocortical adenomas (three cortisol-producing adenomas and one aldosterone producing adenoma) and normal adrenal tissues. The effect of epinephrine on cortisol production was also evaluated in vitro from the cultured tumor cells from one cortisol-producing adenoma. Saturable binding of [3H]DHA to the tumor membranes was observed in two of three cortisol-producing adenomas, but not in the aldosterone-producing adenoma or in normal adrenal tissues. Scatchard analysis of equilibrium binding of [3H]DHA revealed a single class of binding sites on the tumor membranes; the apparent dissociation constant (Kd) was 1 nM in each, and the numbers of binding sites were 108 and 45 fmol/mg protein, respectively. Competition by adrenergic agents with [3H]DHA for binding sites on the membranes from one cortisol-producing adenoma revealed that (+/-)propranolol, a beta-adrenergic antagonist, was about 350-fold more potent than phentolamine, an alpha-adrenergic antagonist, suggesting the beta-adrenergic nature of receptor sites. In addition, stereospecificity was demonstrated by about 1000-fold greater affinity of (-)alprenolol than to (+)alprenolol, both of which are stereoisomers of the beta-adrenergic antagonist. Furthermore, production of cortisol from the cultured tumor cells prepared from the same adenoma was significantly stimulated by epinephrine in addition to ACTH. These data indicate that ectopic beta adrenergic receptor sites are present in some human adrenocortical tumors which may be functionally related to the activation of adenylate cyclase by catecholamines other than ACTH in those tumors, as previously demonstrated. The mechanism by which such altered cellular membrane characteristics occur in association with neoplastic alteration of the endocrine tissues remains unanswered. PMID- 6270186 TI - The zonal origins of the mineralocorticoid hormones in the 21-hydroxylation deficiency of congenital adrenal hyperplasia. AB - The 0800 h plasma concentrations of the mineralocorticoid hormones, 18 hydroxydeoxycorticosterone (18-OHDOC), deoxycorticosterone (DOC), corticosterone, 18-hydroxycorticosterone (18-OHB), and aldosterone, in six patients with nonsalt losing congenital adrenal hyperplasia revealed two groupings of these steroids: in one group, DOC, 18-OHB, and aldosterone were significantly elevated (P less than 0.001) at 51.7 +/- 18.0, 70.8 +/- 14.2, and 22.7 +/- 3.0 ng/dl, respectively; in the other group, corticosterone and 18-OHDOC were normal at 363.6 +/- 76.0 and 7.8 +/- 1.1 ng/dl, respectively. No significant increases in response to upright posture were observed in DOC, 18-OHB, or aldosterone. After a 1-h Cortrosyn stimulation test, the already elevated levels of DOC, 18-OHB, and aldosterone showed slight additional increases, but the normal levels of corticosterone and 18-OHDOC changed little within the normal unstimulated range. In these patients certain mineralocorticoid hormone patterns permit the identification of the zonal origins of steroids. The normal and fixed levels of 18-OHDOC and corticosterone, zona fasciculata steroids, are similar to those of cortisol and imply deficiency of formation and of their precursor, zona fasciculata DOC, a 21-hydroxylated steroid. Both the mineralocorticoid and glucocorticoid pathways distal to 21-hydroxylation are impaired in the zona fasciculata. However, the elevated and partially responsive levels of DOC, 18 OHB, and aldosterone imply that there is greater activation of 21-hydroxylation in the zona glomerulosa than in the zona fasciculata, with its normal fixed steroid levels, and that the elevated level of DOC is primarily from this zone. PMID- 6270187 TI - Characterization of calcitonin- and adrenocorticotropin-producing human cloned cell lines. AB - We have developed three human cloned cell lines that produce immunoreactive human calcitonin (ihCT) and ACTH (iACTH) as well as exhibit characteristics of cultured neural cells. Clones HMS-41/I, -78/2, and -98/2 were developed from cell lines HeLa AV3, MBA 9812 (bronchogenic carcinoma), and SW 267 (pheochromocytoma), respectively. Karyological analysis of both the parent and the cloned cell lines confirmed the identity of HeLa AV3 and MBA 9812. When grown in serum-free media designed for culturing neural cells, the patterns of production for both ihCT and iACTH varied among the clones. The multiple patterns of hormone production suggest that the mechanisms involved in the biosynthesis, processing, and secretion of these hormones differ among the clones. The clones contain neuron specific enolase and the putative neurotransmitters beta-alanine and gamma-amino butyric acid, and they respond to cAMP analogs by differentiating, as noted by the extension of neurites (except the HeLa-derived HMS-41/I). The iACTH extracted from cells and synthetic ACTH exhibited equivalent profiles upon isoelectric focusing. The forms of ihCT noted in cell extracts were similar to those observed in extracts of human tumor tissue. Our rabbit antiserum to hCT failed to detect ihCT in those cell extracts prepared for ACTH determination or in extracts of rat pituitaries, but it did detect CT in rat thyroids by both RIA and immunofluorescent procedures. We concluded that our antisera to hCT do not detect the precursor form of ACTH. The availability of these cloned cell lines provides model systems for examining the production of these peptide hormones and the concomitant expression of neural and endocrine characteristics. PMID- 6270188 TI - Calcitonin-like immunoreactivity in the pituitary vascular bed of man. AB - The calcitonin-like immunoreactivity (CT-like immunoreactivity) was measured in blood aspirated from the vascular bed of the anterior pituitary gland during transsphenoidal surgery in 33 patients with PRL-producing microadenomas, 2 patients with Cushing's disease, and 1 patient with metastatic breast cancer with a normal pituitary gland. The mean level of CT-like immunoreactivity in the pituitary vascular bed was 2-3 times higher than in peripheral blood (2.4 +/- 0.9 ng/ml vs. 0.69 +/- 0.19 ng/ml), and the difference was highly significant (P less than 0.001). However, the serum ACTH, hGH, TSH, PRL, and FSH in the pituitary vascular bed was 1000 times or higher than that found in the peripheral blood. The serum CT-like immunoreactivity levels in the pituitary bed in the two patients with Cushing's disease were similar to that found in other patients. Our investigations indicate: 1) CT-like immunoreactivity in man is higher in the blood obtained from the pituitary vascular bed than that found in the peripheral blood; 2) the serum CT-like immunoreactivity level in the pituitary vascular bed is much less than ACTH or the other hormones secreted by the pituitary gland; 3) there is no correlation between CT-like immunoreactivity and ACTH levels. PMID- 6270189 TI - Difference in antibody reactivity between complement fixation and immune adherence hemagglutination tests with virus antigens. AB - Complement-fixing (CF) immunoglobulin M antibody to infantile gastroenteritis virus (a rotavirus) did not show the reactivity of immune adherence hemagglutination (IAHA). Early immunoglobulin G CF antibody produced both in patients and in guinea pigs experimentally infected with Japanese encephalitis virus (a flavivirus) had weak reactivity in IAHA test. However, late antibody showed higher titers by IAHA than by CF. These results suggested that early antibodies with lower affinity are inefficient in the IAHA reaction. The implications of this study are: (i) a low ratio of IAHA/CF antibody titers in a serum suggests a recent rotavirus infection; (ii) the IAHA reaction is more type specific than the CF reaction for identifying the serotype of antigenically cross reacting viruses with hyperimmune sera. PMID- 6270190 TI - Rapid, simple method of preparing rotaviral double-stranded ribonucleic acid for analysis by polyacrylamide gel electrophoresis. AB - A procedure for extracting rotaviral double-stranded ribonucleic acid (RNA) directly from fecal and intestinal specimens collected from calves and pigs is described. This procedure provides a rapid, simple, reproducible method of obtaining rotaviral double-stranded RNA preparations suitable for electrophoretic analysis in polyacrylamide-agarose composite gels. The rotaviral genome electrophoretic migration pattern produced by double-stranded RNA extracted directly from a specimen by this procedure was qualitatively identical to the electrophoretic migration pattern obtained with double-stranded RNA extracted from purified rotavirus derived from the same specimen. Direct extraction of specimens containing porcine rotavirus-like virus by this procedure gave preparations that had electrophoretic migration patterns similar, but not identical, to the characteristic electrophoretic migration pattern of the rotaviral genome. Sufficient rotaviral double-stranded RNA could be extracted from 6 ml of fecal or intestinal specimen by this procedure to permit 15 or more electrophoretic assays. PMID- 6270191 TI - Enzyme immunoassays for measurement of cytomegalovirus immunoglobulin M antibody. AB - The diagnosis of congenital cytomegalovirus (CMV) infection is often accomplished by the detection of circulating antibody directed against CMV. We devised a method for measuring CMV-specific immunoglobulin M (IgM) based on the isolation of IgM antibody by reaction with a solid phase coated with antihuman IgM. The determination of IgM antibody specific for CMV was accomplished by the subsequent addition of CMV or control antigen and enzyme-labeled CMV antibody (solid phase IgM method). We compared the sensitivity and specificity of this method with those of a conventional form of solid-phase enzyme immunoassay in which CMV antigen is bound to the solid phase (solid phase-antigen method). Both assay systems were capable of detecting CMV-specific IgM antibody in the sera of 10 babies with documented CMV infection and in those of the mothers of 4 of these babies. The solid phase-IgM method yielded negative results in all 66 sera available from babies who did not have congenital CMV infection. On the other hand, the solid phase-antigen system yielded false-positive results in 12 (18%) of these sera. In addition, the solid phase-antigen system yielded false-positive results in 8 of 12 sera obtained from patients with demonstrable rheumatoid factor. However, the solid phase-IgM system yielded negative results for the rheumatoid sera, provided that appropriate control reactions were performed. The solid phase-IgM system is thus a specific and sensitive method for the determination of CMV IgM antibody. PMID- 6270192 TI - Direct effects of thyrotropin-releasing hormone, cyproheptadine, and dopamine on adrenocorticotropin secretion from human corticotroph adenoma cells in vitro. AB - In an attempt to delineate the mechanism and the site of action of cyproheptadine and dopaminergic agonists as well as hormones including thyrotropin-releasing hormone (TRH) and hydrocortisone, the effects of these substances on ACTH secretion from corticotroph adenoma cells in culture were examined. Dispersed cells of pituitary adenomas obtained at surgery from four patients with Nelson's syndrome and one subject with Cushing's disease formed a monolayer and actively secreted ACTH into the medium. When TRH (0.1 microM) was added to the medium, a significant increase in ACTH secretion was demonstrated by adenoma cells from two patients who responded to TRH preoperatively. Moreover, a dose-response relationship between TRH concentrations and ACTH secretion was observed. Incubation of cells with cyproheptadine (1 or 0.1 microM) resulted in a significant decrease in ACTH release, and inhibited stimulation produced by TRH in one experiment. This effect of cyproheptadine was blocked when equimolar concentrations of serotonin was coincubated, whereas serotonin by itself did not affect ACTH secretion. Dopamine (0.1 microM) lowered ACTH accumulation in the medium, which was blocked by the addition of haloperidol. When hydrocortisone was added to the culture, dose-dependent suppression of ACTH secretion was demonstrated. TRH at an equimolar concentration reversed this effect, but, failed to overcome the inhibition induced by a higher concentration of hydrocortisone in cells from one adenoma studied. Cultured normal corticotrophs obtained from a patient with metastatic breast cancer, on the other hand, did not show any response to these substances, except for hydrocortisone. We suggest that TRH, cyproheptadine, dopamine affect ACTH secretion in patients with ACTH-producing pituitary adenomas by their direct action on the adenoma. PMID- 6270193 TI - Effect of quinidine on the digoxin receptor in vitro. AB - To investigate the basis for a clinically important digitalis-quinidine interaction that is characterized by increases in serums digoxin concentrations when quinidine is administered to digoxin-treated patients, we have studied in vitro the interaction of quinidine with the digoxin receptor. Evidence has been obtained that quinidine is capable of decreasing the affinity for digoxin of cardiac glycoside receptor sites on purified Na,K-ATPase and on intact human erythrocyte membranes. As others have shown, quinidine is capable of inhibiting Na,K-ATPase activity, and evidence has been obtained in the current study that, while quinidine can reduce the affinity of the enzyme for digoxin, it is also capable of acting together with digoxin in inhibiting enzyme activity to a degree greater than the inhibitory effect of digoxin alone. The concentrations of digoxin and quinidine used in this study were considerably greater than their therapeutic serum concentrations. Nevertheless, these observations are consistent with the hypothesis that the increases in serum digoxin concentrations and the decreases in volumes of digoxin distribution observed clinically when quinidine is administered to digoxin-treated patients may reflect, at least in part, a decrease in the affinity of tissue receptors for digoxin. The possibility must also be considered that enhanced cardiac effects of digoxin may occur clinically as the result of an augmentation, by quinidine, of digoxin effects, which more than compensates for the modest reduction in digoxin binding. PMID- 6270194 TI - Familial dysbetalipoproteinemia. Abnormal binding of mutant apoprotein E to low density lipoprotein receptors of human fibroblasts and membranes from liver and adrenal of rats, rabbits, and cows. AB - Patients with familial dysbetalipoproteinemia (F. Dys.), also called familial type 3 hyperlipoproteinemia, are homozygous for a mutant allele, Ed, that specifies an abnormal form of apoprotein (apo) E, a prominent constituent of remnant lipoproteins derived from very low density lipoproteins (VLDL) and chylomicrons. Apo E is thought to mediate the removal of remnant lipoproteins from the plasma by virtue of its ability to bind to hepatic lipoprotein receptors. In F. Dys. patients, remnant-like lipoproteins accumulate, apparently because of delayed clearance by the liver. In the current studies, we show that the abnormal protein specified by the Ed allele (apo E-D) from some, but not all, patients with F. Dys. has a markedly deficient ability to bind to low density lipoprotein (LDL) receptors. Apo E was isolated from eight control subjects and nine patients with F. Dys. and incorporated into phospholipid complexes. The complexes were tested for their ability to compete with human 125I-LDL or rabbit 125I-beta-VLDL fo binding to LDL receptors in four assay systems: cultured human fibroblasts, solubilized receptors from bovine adrenal cortex, liver membranes from rats treated with 17 alpha-ethinyl estradiol, and liver membranes from normal rabbits. The apo E-D from six of the nine patients with F. Dys. showed binding affinities for LDL receptors that were reduced by greater than 98% in all receptor assays (group 1 patients). All of these group 1 patients were unequivocally of phenotype apo E-D/D by the criterion of isoelectric focussing. The apo E from the three other F. Dys. patients showed a near normal binding ability in all four of the receptor assays (group 2 patients). One of these group 2 patients appeared to have the apo E-D/D phenotype by isoelectric focussing. In the other two patients in group 2, apo E-D was the predominant protein (phenotype, apo E-D/D), but traces of protein in the region corresponding to normal apo E (apo E-N) were also present. The difference between group 1 and group 2 patients was also apparent when the apo E was iodinated and tested directly for binding to liver membranes from rats treated with 17 alpha-ethinyl estradiol. The 125I-labeled apo E from a group 2 patient, but not a group 1 patient, showed enhanced uptake when perfused through the liver of an estradiol treated rate, indicating that the receptor binding ability of apo E correlated with uptake in the intact liver. The current studies allow the subdivision of patients with F. Dys. into two groups. In group 1, the elevated plasma level of remnants appears to be due to a diminished receptor binding activity of the abnormal protein specified by the Ed allele; in group 2 patients, the cause of the elevated plasma level of remnants remains to be explained. PMID- 6270195 TI - A new polymorphism in the human beta-globin gene useful in antenatal diagnosis. AB - A new polymorphism in the beta-globin is described, using the restriction enzyme Asu I. A radioactive probe specifically representing the large intervening sequence (IVS 2) of the beta-globin gene has been used to detect this polymorphism. Normally, a 0.8-kilobase fragment containing beta-IVS 2 is generated by Asu I; however, a 1.0-kilobase fragment is seen in association with 18% of beta A-genes, and 38% of beta-thalassemia genes in an Israeli population studied. By contrast, the Asu I polymorphism has rarely been seen in blacks examined to date. An additional Asu I change is seen the the delta-globin gene with a delta-IVS probe. The beta-Asu I polymorphism is shown to be useful in the antenatal diagnosis of beta-thalassemia. PMID- 6270197 TI - Pigeon herpes infection: excretion and re-excretion of virus after experimental infection. PMID- 6270196 TI - A role for prostaglandins and thromboxanes in the exposure of platelet fibrinogen receptors. AB - Exposure of fibrinogen receptors by a variety of agonists is a prerequisite for platelet aggregation. Because the synthesis of prostaglandins and thromboxane A2 also occurs during platelet aggregation we wondered whether these agents participate in the exposure of platelet fibrinogen receptors. Therefore, we measured the binding of human 125I-fibrinogen to gel-filtered normal human platelets after prostaglandin and thromboxane synthesis had been inhibited by aspirin or indomethacin. The fibrinogen binding assay was performed at 37 degrees C but without stirring to prevent the formation of platelet aggregates. Platelet secretion, measured with [14C]serotonin, did not occur during the procedure. Aspirin or indomethacin inhibited fibrinogen binding stimulated by 10 microM epinephrine by 53%, and inhibited fibrinogen binding stimulated by 1-2 microM ADP by 37.1%. However, ADP at concentrations greater than 2 microM returned fibrinogen binding toward control values. Scatchard analysis demonstrated that aspirin decreased the number but not the affinity of the exposed fibrinogen receptors. To determine whether prostaglandins are capable of directly exposing fibrinogen receptors, prostaglandin H2 was used to stimulate platelets in the fibrinogen binding assay. Prostaglandin H2 exposed approximately 54,000 fibrinogen receptors/platelet and corrected the deficit in receptor exposure induced by aspirin. These studies demonstrate that platelet prostaglandins or thromboxane A2 can play a direct role in the exposure of platelet fibrinogen receptors. In addition, they suggest that the synthesis of prostaglandins and thromboxane A2 by stimulated platelets may be all that is required for optimal secondary platelet aggregation. PMID- 6270198 TI - Pigeon herpes infection: natural transmission of the disease. PMID- 6270199 TI - Theories and predictions of models describing sequential interactions between the receptor, the GTP regulatory unit, and the catalytic unit of hormone dependent adenylate cyclases. PMID- 6270200 TI - Comparison of cAMP-dependent protein kinase in normal and Rous sarcoma virus transformed chick embryo fibroblasts. AB - cAMP-dependent protein kinase was compared in normal and Rous Sarcoma Virus transformed chicken embryo fibroblasts. Total cAMP binding activity and cAMP dependent histone kinase activity were unaltered by RSV transformation. The apparent Km for activation of histone kinase activity by cAMP was 35 nM in both normal and transformed cells. Using 8-N3-cAMP photoaffinity labeling, normal and transformed cells were also found to contain equal quantities of a single 42,000 Mr regulatory sub-unit isoenzyme of A-kinase. This isoenzyme corresponded to the lower molecular weight isoenzyme of the two enzymes found in normal chicken skeletal muscle. Both avian isoenzymes were about 4,000 Mr smaller than the corresponding bovine type I and type II regulatory subunits. Rous Sarcoma Virus transformation does not directly alter the amount or activity of cAMP-dependent protein kinase. PMID- 6270201 TI - A case of porocarcinoma. PMID- 6270202 TI - Influences on medical education in the United States. PMID- 6270203 TI - Training female mental health professionals: sexual countertransference issues. PMID- 6270204 TI - Carroll L. Birch Award manuscript: the effects of cadmium on lymphocyte transformation. PMID- 6270205 TI - Syndactylism and its surgical repair. PMID- 6270206 TI - Autoradiographic determination of neurotransmitter receptor distributions in the cerebral and cerebellar cortices. AB - Light microscopic autoradiographical localization of drug and neurotransmitter receptors provides investigators with a tool to determine the sites of drug action with a high degree of anatomical resolution. In the cortex, where there is a distinct laminar organization, the differential distribution of receptors can be determined. We have found that differences in the density of specific receptors exist between layers of the cerebral cortex. Our observations include the localization of muscarinic cholinergic, benzodiazepine, opiate, neurotensin, histamine-H1, alpha-adrenergic, beta-adrenergic, and high-affinity GABA receptors in the cerebral cortex. Of this list of neurotransmitters only significant concentrations of benzodiazepine, GABA, and beta-adrenergic receptors were found in the cerebellar cortex. Receptor localizations such as these can be coupled with information from immunohistochemical studies to provide the basis for future experimentation in order to define neurotransmitter-specific pathways in the cerebral and cerebellar cortices. PMID- 6270207 TI - Autoradiography of diffusible substances: verification of the specificity of the localization by correlated physiological, biochemical, and pharmacological studies. AB - Recent advances in autoradiographic technique make it possible to localize quite a diverse class of soluble and/or diffusible substances. However since the molecules in question could move or be displaced at any time during preparation it is necessary to employ chemical or physiological tests to prove the validity of the localization. For two classes of compounds this can be done quite readily. 3H-ouabain, a cardiac glycoside that binds specifically to the Na pump, can be monitored for its effect on Na transport and the binding itself can be controlled by manipulating specific ligands known to modulate the ouabain-Na pump interaction. Acetylcholine can be readily analyzed by chemical means and its solubility characteristics allow one to extract it even from osmium-fixed embedded sections. Thus it is possible to monitor for the presence of acetylcholine in tissue through the processing for autoradiography and even chemically characterize small areas of tissue by microdissection of sectioned material. PMID- 6270208 TI - A new species of lysosome in rabbit polymorphonuclear leukocytes. AB - A species of lysosome containing an acid hydrolase activity, i.e., acid trimetaphosphatase (TMPase) was demonstrated in rabbit polymorphonuclear leukocytes (PMNs). The lysosome was less reactive near neutral pH and non reactive at alkaline pH. The structure appeared typically ovoid, unlike the "tubular lysosomes" reported by Oliver (J Histochem Cytochem 28:78, 1980) in several exocrine acinar cells and also observed in monocytes in this study. Neither was there Golgi activity, although there are probably few, if any, Golgi elements present in the mature neutrophil. It is interesting that the TMPase containing lysosome occurs in an inclusion the size of a specific granule at a greater frequency than the nonspecific acid phosphatase-containing lysosome. PMID- 6270209 TI - Virus diarrhoea associated with pale fatty faeces. AB - Steatorrhoea was a significant feature in an outbreak of rotavirus gastroenteritis which affected adults and infants in hospital. Fat globules or fatty acid crystals were obvious by light microscopy (LM) in faeces from 14 of 25 patients examined. Ten of the fatty stools and two of the remainder were very pale. By electron microscopy (EM) a rotavirus was seen in 11 of the 14 fatty faeces and in only two of 11 specimens without visible fat. In a further study of pale or fatty faeces 20 such specimens sent for laboratory examination from patients not involved in the hospital outbreak were compared microbiologically with a similar number which were neither pale nor fatty. Viruses were found by EM in 11 (55%) of the pale or fatty stools; eight rotaviruses, two astroviruses and an uncultivable adenovirus were seen; one further patient had acute jaundice. In contrast, no viruses were seen by EM in the twenty specimens which were normally pigmented and without evident fat. Steatorrhoea was significantly associated with rotavirus infection of the alimentary tract which usually presented as a fatty enteritis. We conclude that rotaviruses certainly, and other viruses possibly, can impede both the digestion of fat and the pigmentation of the faeces. Inspection and LM of faeces are easy. In acute enteritis a fatty or pale stool is an indication for virological examination. PMID- 6270210 TI - Immunofluorescent staining of nuclear antigen in lymphoid cells transformed by Herpesvirus papio (HVP). AB - An improved fixation method for antigen detection in lymphoblastoid cells is described. Herpesvirus papio nuclear antigen (HUPNA) could be stained in several transformed lymphoid cell lines by anti-complement immunofluorescence (ACIF). Antibody to HUPNA was detected in many human sera containing antibodies to Epstein-Barr virus capsid and nuclear antigen (EBNA). Rheumatoid arthritis sera showed a high incidence of both anti-EBNA and anti-HUPNA antibodies. PMID- 6270211 TI - Use of chelating agents as terminators of alkaline phosphatase activity in enzyme linked immunosorbent assay (ELISA) tests. AB - For a long time chelating agents have been known to be inhibitors of alkaline phosphatase activity. However, the use of chelating agents in stopping alkaline phosphatase activity in ELISA reactions is a novel application with several advantages over conventionally used acids or bases. PMID- 6270212 TI - Simultaneous isolation of bovine eosinophils and neutrophils on gradients of Percoll. AB - Bovine eosinophils and neutrophils have been isolated from venous blood using a two-step procedure which involves initial preparation of a granulocyte-rich fraction by centrifugation and osmotic shock followed by sensity gradient centrifugation on Percoll. Good yields of both cell types are obtained with purities of 90% or better. The isolated cells are viable and actively phagocytic. The effects of phagocytosis on the metabolism of carbon-1 of d-glucose and the incorporation of iodine have been measured and several differences between bovine cells and those of other species were noted. PMID- 6270213 TI - Activation of cAMP-dependent protein kinase in epidermis by the compounds which increase epidermal cAMP. AB - Pig epidermal slices were incubated with various compounds which increased epidermal cAMP (adenosine 3',5'-monophosphate), and the change in cAMP-dependent protein kinase activity ratio was studied by the method of Cherrington et al (J Biol Chem 251:5209-5218, 1976) with modification. Epinephrine (5 x 10(-5) M), histamine (10(-4) M) and adenosine (10(-3) M), potent agonists of epidermal adenyl cyclase, fully activated the protein kinase (PK) during an incubation of 30 to 45 seconds, that was much shorter than that required for maximal cAMP accumulation under the same conditions (5 min). With such a brief stimulus, the epidermal cAMP-PK system did not become refractory and responded to repeated stimuli. Prostaglandin E2 (PGE2) and isobuthylmethylxanthine (IBMX) and ethanol only partially activated the enzyme. Prostaglandin F2 alpha (PGF2 alpha) and theophylline which were much less effective in increasing epidermal cAMP, activated the enzyme to the same extent as PGE2 and IBMX respectively. These results suggest that protein kinase activation takes place in response to a cAMP increase in small locus of the cell. Such an increase in cAMP can be very small or even not measurable when measured as total cAMP in the tissue homogenate. Also, increases above this level may not be physiologic. It is concluded that measurement of cAMP-dependent protein kinase activity ratio is a more direct and more sensitive way to study the effect of compounds which act through cAMP mediated mechanisms. PMID- 6270214 TI - Role of myeloperoxidase and bacterial metabolism in chemiluminescence of granulocytes from patients with chronic granulomatous disease. AB - Phagocytosis of catalase-positive microbes by normal polymorphonuclear neutrophils results in increased metabolism as required for microbicidal action. Chemiluminescence is a product of the associated oxygenation reactions. Neutrophils from patients with chronic granulomatous disease are capable of phagocytizing catalase-positive microbes, but there is no associated respiratory burst, microbicidal action is greatly decreased, and chemiluminescence is not detected. However, these defective neutrophils can kill catalase-negative, H2O2 generating bacteria. In the present study, chemiluminescence by neutrophils from patients with chronic granulomatous disease after phagocytosis of H2O2-generating streptococci was detected. Acid extracts of myeloperoxidase from either control or patient neutrophils also yielded chemiluminescence in the presence of streptococci, but not in the presence of catalase-positive microbes. PMID- 6270215 TI - Antistaphylococcal activity of rifampin with other antibiotics. AB - The bactericidal activity of rifampin plus other antibiotics was examined with in vitro checkerboard dilutions against 15 strains of staphylococci, with time-kill studies, and with determinations of serum bactericidal activities (SBAs) in volunteers. In vitro synergism was found consistently only in inhibitory assays with rifampin plus minocycline. Time-kill studies with rifampin plus methicillin revealed enhanced killing at concentrations close to the MBC but reduced combined activity at higher concentrations. SBAs with rifampin, oxacillin, and the two drugs in combination were studied in three treatment regimens (regimen A: 2 g of oxacillin and 300 mg of rifampin; regimen B: 1 g of oxacillin and 450 mg of rifampin; and regimen C: 3 g of oxacillin and 450 mg of rifampin). The SBA against methicillin-sensitive Staphylococcus aureus was significantly lower with the two drugs in combination than with oxacillin alone in regimen A but not in regimen B. In vitro synergy with rifampin plus other antibiotics was not frequent. At high concentrations, the bactericidal activity of oxacillin was reduced by rifampin, but killing activity may be enhanced at low concentrations of oxacillin. PMID- 6270216 TI - Imidazole and polyene activity against chloroquine-resistant Plasmodium falciparum. PMID- 6270217 TI - Evaluation of enzyme immunoassays for the detection of human rotavirus. PMID- 6270218 TI - [Movement of protons and formation of electrochemical proton gradients in oxidative phosphorylation (author's transl)]. PMID- 6270219 TI - [Selective hypoaldosteronism (author's transl)]. PMID- 6270220 TI - [Clinical pictures of cases with protracted course in vibration disease (author's transl)]. PMID- 6270221 TI - [Ectopic hormone-producing tumors (author's transl)]. PMID- 6270222 TI - Concurrent measurement of plasma levels of vitamin D3 and five of its metabolites in normal humans, chronic renal failure patients, and anephric subjects. AB - Here we report the use of newly developed and established techniques for the determination of plasma levels of a broad spectrum of vitamin D3 metabolites, including vitamin D3 and 25OHD3-lactone, in normal humans, chronic renal failure patients, and anephric subjects. The methodology described consisted of methanol methylene chloride extraction, Lipidex-5000 chromatography with stepwise gradient elution, normal-phase HPLC with concave gradient elution, and sensitive ligand binding assays. The results of the study strongly suggest an extrarenal source(s) for 24,25(OH)2D3 and 25,26(OH)2D3 and indicate that both 25OHD3-lactone and 1,25(OH)2D3 may be produced solely in the kidney of the human. Significant reductions or nondetectable plasma levels of vitamin D3 in the renal disease patients may reflect abnormalities in the hepatobiliary-intestinal and/or cutaneous metabolism of vitamin D. PMID- 6270223 TI - [Doppler in the diagnosis of glomus tumours (author's transl)]. PMID- 6270224 TI - Lamellar inclusion bodies in lung-fish thrombocytes. PMID- 6270225 TI - Intracytoplasmic crystals of enterovirus 71 in cultured cells. PMID- 6270226 TI - Mast cell mediator release as a function of cyclic AMP-dependent protein kinase activation. AB - Stereo-specific perturbation of the IgE-receptor (shown in previous studies) produces a monophasic rise in cyclic AMP that peaks at 15 s and a depletion of cyclic AMP-dependent protein kinase that plateaus at 30-60 s. The previously observed linear relationship between the attenuation in the monophasic rise in cyclic AMP and the quantity of mediator release in the presence of incremental concentrations of the adenosine analogue 2',5',-dideoxyadenosine, DDA, which is known to inhibit adenylate cyclase, indicated a direct relationship between receptor perturbation, transmembrane activation of adenylate cyclase, and granule secretion. The role of cyclic AMP as a second messenger in this sequence is now apparent from the linear relationship between net percent mediator release and net percent activation of cyclic AMP-dependent protein kinase isoenzyme when IgE dependent activation of adenylate cyclase is suppressed by incremental quantities of DDA. There was a comparable percent activation of both types I and II mast cell cyclic AMP-dependent protein kinase isoenzymes with anti-IgE-induced activation and secretion, and there was a parallel suppression of the activation of both isoenzymes in the presence of DDA. Although these studies firmly link the activation of cytoplasmic cyclic AMP-dependent protein kinase to the IgE receptor initiated transmembrane activation of adenylate cyclase. they do not discriminate among the functions of the two isoenzymes. PMID- 6270227 TI - Induction of monocyte procoagulant activity by murine hepatitis virus type 3 parallels disease susceptibility in mice. AB - The in vitro induction of procoagulant activity (PCA) in murine peripheral blood mononuclear cells (PBM) by murine hepatitis virus type 3 (MHV-3) correlates with the disease susceptibility in three strains of mice. PBM from BALB/c mice, a strain in which MHV-3 infection results in fatal acute fulminant hepatitis, responds to the virus with a robust PCA response, whereas PBM from C3H/St mice, a strain which develops mild acute hepatitis followed by chronic hepatitis, only exhibit a modest PCA response. In contrast, PBM from A/J mice, a strain fully resistant to MHV-3, generate no increase in PCA above control levels. The induction phase of MHV-3 PCA is rapid, with an increase within 1-1.5 h, with maximum activity at 18h, and it precedes MHV-3 replication in either 17 CL1 cells, a fully permissive cell line, or in monocytes from these strains of mice. The PCA response of BALB/c PBM exceeds the response to any other known stimulus. No induction occurs upon direct stimulation of monocytes by MHV-3, but in the presence of lymphocyte collaboration, the PCA response is observed first at a lymphocyte:monocyte ratio of 2:1 and reaches a maximum as the lymphocyte:monocyte ratio approaches 4:1. This response appears to provide a functional marker for susceptibility to MHV-3 infection in inbred strains of mice and could be important in the pathogenesis of MHV-3-induced disease. PMID- 6270228 TI - Genetics of susceptibility for radiation-induced leukemia. Mapping of genes involved to chromosomes 1, 2, and 4, and implications for a viral etiology in the disease. AB - Susceptibility to radiation-induced leukemia in (A/J x B10)F2 mice is encoded for by genes in chromosomes 1, 2, and 4. The loci involved in chromosomes 1 and 4 are close to or similar to xenotropic virus inducibility locus on chromosome 1 and a locus-affecting expression of xenotropic MuLV envelope-related cell surface antigens. Radiation-induced leukemia-1 (Ril-1) on chromosome 2 plays an overriding influence in susceptibility to the disease. This locus might encode ecotropic viral-associated genetic information or might contain cellular sequences with oncogenic potential. These findings are of interest in view of the importance of recombinant viruses to leukemogenesis. Furthermore, it is intriguing that Ril-1 is located in a chromosomal site rich in thymus differentiation-specific loci. An explanation for tissue-specific activation of endogenous viruses is that activation of the virus in question is dependent on differentiation-specific steps. PMID- 6270229 TI - Functional characterization of synthetic leukotriene B and its stereochemical isomers. AB - Leukotriene B (LTB), a potent lipid chemotactic factor for neutrophils, is 5S,12R dihydroxy-6,14-cis,8,10-trans-eicosatetraenoic acid (Fig 1), based upon direct comparison of natural LTB with synthetic 5S,12R-dihydroxy-6,8,10,14 eicosatetraenoic acid (5,12-di-HETE) stereoisomers in three biological assays. Of the six synthetic stereoisomers evaluated, only the 5S,12R,6,14-cis,8,10-trans compound had chemotactic potency for human neutrophils in vitro that was comparable to that of natural LTB, with a concentration of 3 X 10(9-9) M eliciting a one-half maximum response. In contrast, the racemic mixture of 5R,12R and 5S,12S-6,10-trans,8,14-cis, the racemic mixture of 5S,12R- and 5R,12S-6,10 trans,8,14-cis, the 5S,12R-6,8-trans,10,14-cis, the 5S,12R-6,8,10-trans,14-cis, and the 5S,12S-6,8,10-trans,14-cis stereoisomers required concentrations of 3 X 10(-7) to 1 X 10(-6) M to elicit comparable responses. Only natural LTB and its synthetic counterpart elicited a local neutrophil infiltration when injected into the skin of the rhesus monkey at 10 ng and 100 ng per site. Natural and synthetic LTB at a concentration of 3 X 10(-8) M each provoked an EC25 contractile response of guinea pig pulmonary parenchymal strips in vitro, whereas the other four tested stereoisomers of 5,12-di-HETE were inactive at this concentration. Structure-function analyses suggest that the neutrophil chemotactic activity depends critically upon the C-1 to C-12 domain, including the stereochemistry of the 6-,8-,and 10-olefinic bonds and the presence of both hydroxyl groups. PMID- 6270230 TI - Short- and long-term decrements in toxicosis-induced odor-aversion learning: the role of duration of exposure to an odor. AB - Six experiments employed an odor-aversion paradigm to investigate the role of the duration of exposure to an odor in determining that odor's subsequent associability with illness. Rats were exposed to an odor at times T1 and T2, and the second of these exposures was followed by toxicosis. When the initial odor exposure was brief, the odor aversion was attenuated with a moderate T1-T2 interval of 3 hr (Experiment 1) but not with long intervals of 28 hr and 76 hr (Experiment 2). In contrast, when the initial odor exposure was long, the odor aversion was attenuated at a long T1-T2 interval (Experiment 3). With a T1-T2 interval of 24 hr, a brief initial exposure did not attenuate odor aversions when the context either remained the same or was changed from T1 to T2, whereas a long initial exposure attenuated such aversions when the context remained the same but not when the context was changed (Experiment 4). With a T1-T2 interval of 3 hr, a brief initial exposure attenuated odor aversions when the context remained the same or was changed from T1 to T2, whereas a long initial exposure attenuated such aversions when the context remained the same but not when the context was changed (Experiment 5). A brief exposure at T1, either with or without a subsequent context "extinction," attenuated odor aversions when the T1-T2 interval was 3 hr but not when this interval was 24 hr; a long initial exposure at T1, without a subsequent context "extinction," attenuated odor aversions when the T1-T2 interval was 4 hr and 24 hr but with a subsequent context "extinction" did not attenuate such aversions at either 4-hr or 24-hr T1-T2 intervals (Experiment 6). The results demonstrate that the duration of exposure to an odor determined whether that odor presentation caused short- or long-term decrements in odor conditionability and are discussed in terms of the relation between self- and retrieval-generated processes. PMID- 6270231 TI - A comparative study of the activity of lysosomal and main metabolic pathway enzymes in tissue biopsies and cultured fibroblasts from Dupuytren's disease and palmar fascia. On the pathobiochemistry of connective tissue proliferation, I. AB - Activities of ten main metabolic pathway enzymes and seven lysosomal enzymes were determined in specimens from human normal palmar fascia and Dupuytren's contracture. The activities of the enzymes tested are 2-3 times higher in fresh specimens of Dupuytren's contracture. There are no differences in the activity distribution patterns of both these specimens, or in the absolute activities calculated in relation of the glyceraldehyde-3-phosphate dehydrogenase activities. With the exception of adenylate kinase and pyruvate kinase, the activities of main metabolic pathway enzymes based on the DNA content showed significantly lower activities in Dupuytren's contracture tissue than in palmar fascia. Lysosomal enzymes exhibit no significant differences of activity in the respective specimens. However, the lysosomal enzyme activities of cultured fibroblasts are lower than the corresponding activities from tissue specimens. The enzyme activities per DNA content in cultured fibroblasts are 10-50 times higher than in tissue specimens. The enzyme activities in cultured fibroblasts decrease with age or density of the cells in culture. The increased metabolic activity of the diseased tissues in Dupuytren's contracture is due to the higher cell content of the afflicted portions of the tissue, but individual enzymes show no qualitative changes in activity and there are no increases of enzyme activity per cell (DNA). PMID- 6270232 TI - Urinary excretion of cyclic guanosine 3'.5'-monophosphate in children with malignant tumours. AB - The present study shows an increased urinary cyclic guanosine 3'.5'-monophosphate (cyclic GMP) excretion rate in children of all age groups bearing malignant tumours or lymphomas. The incidence of increased cyclic GMP excretion was highly significant (79%). Follow-up studies of up to three years have revealed that during periods of remission of malignant disease the urinary cyclic GMP excretion drops to near normal values, whereas recurrences are accompanied by a new increase of cyclic GMP excretion. PMID- 6270233 TI - Cyclic AMP in mammalian follicle cells and oocytes during maturation. AB - Intact ovarian follicles and isolated oocytes were cultured for 30 sec to 18 hr in the presence of gonadotrophins, and the cyclic AMP content in the follicle and oocyte was measured. The basal content of cyclic AMP in ovine oocytes before gonadotrophic stimulation was 6.3 +/- 0.7 fmol/oocyte or 8 microM. There was no fall in oocyte cyclic AMP concentration as an immediate response to the gonadotrophins, but at 1, and 12-18 hr after stimulation, the concentration in both the oocytes and follicle cells was considerably elevated. There was no comparable increase in intracellular cyclic AMP in oocytes denuded of follicle cells before culture, even when both gonadotrophins and phosphodiesterase inhibitors were included in the medium. We conclude that the signal which initiates oocyte maturation in mammals differs from that of amphibia, where an early fall in intracellular cyclic AMP is essential for the resumption of meiosis. Moreover, cellular interactions within the mammalian follicle are necessary for the characteristic periods of increased cyclic AMP in oocytes during maturation. PMID- 6270234 TI - [Neural tumors of the nasal fossae]. PMID- 6270235 TI - Tetrodotoxin block of sodium channels in rabbit Purkinje fibers. Interactions between toxin binding and channel gating. AB - Tetrodotoxin (TTX) block of cardiac sodium channels was studied in rabbit Purkinje fibers using a two-microelectrode voltage clamp to measure sodium current. INa decreases with TTX as if one toxin molecule blocks one channel with a dissociation constant KD approximately equal to 1 microM. KD remains unchanged when INa is partially inactivated by steady depolarization. Thus, TTX binding and channel inactivation are independent at equilibrium. Interactions between toxin binding and gating were revealed, however, by kinetic behavior that depends on rates of equilibration. For example, frequent suprathreshold pulses produce extra use-dependent block beyond the tonic block seen with widely spaced stimuli. Such lingering aftereffects of depolarization were characterized by double-pulse experiments. The extra block decays slowly enough (tau approximately equal to 5 s) to be easily separated from normal recovery from inactivation (tau less than 0.2 s at 18 degrees C). The amount of extra block increases to a saturating level with conditioning depolarizations that produce inactivation without detectable activation. Stronger depolarizations that clearly open channels give the same final level of extra block, but its development includes a fast phase whose voltage- and time-dependence resemble channel activation. Thus, TTX block and channel gating are not independent, as believed for nerve. Kinetically, TTX resembles local anesthetics, but its affinity remains unchanged during maintained depolarization. On this last point, comparison of our INa results and earlier upstroke velocity (Vmax) measurements illustrates how much these approaches can differ. PMID- 6270236 TI - In situ electron microscopical observation of cells infected with herpes simplex virus. AB - Transport and release of herpes simplex virus (HSV) in an African green monkey kidney cell line (CV-1) was followed by electron microscopy for up to 24 h p.i. Transmission electron microscopy and scanning electron microscopy were employed. For the former approach, electron microscopical autoradiography of whole cultured cells and in situ thin section techniques were used. The following new observations were made. (1) Except in peripheral parts of the cells, where the cytoplasmic membrane could make a ruffling movement relatively freely, virus particles were found only on the dorsal surface of the cell and not on the surface facing the substratum. By observation of thin section in situ, it was confirmed that the virus particles within intracytoplasmic vacuoles were apparently released by a reverse phagocytic process from the cell surface adjacent to microvillus projections. (2) Progeny virus particles in the nucleus moved to the cell surface within 2 h after maturation. PMID- 6270237 TI - Detection of simian virus 40 T-antigen-related antigens by a 125I-protein A binding assay and by immunofluorescence microscopy on the surface of SV40 transformed monolayer cells. AB - Simian virus 40 (SV40)-transformed cells express the SV40-specific tumour transplantation antigen (TSTA) on the cell surface and the SV40-coded tumour antigen in their nuclei. TSTA is defined by SV40-specific transplantation immunity, whereas T-antigen (T-Ag) can be detected serologically by indirect immunofluorescence. Both antigens, however, are derived from the A gene of SV40. We therefore analysed SV40-transformed cells for the presence of serologically detectable T-Ag-related molecules. Such antigens could not be detected on the surface of living SV40-transformed cells in monolayers. However, after a short formaldehyde fixation it was possible to stain the cell surfaces of SV40 transformed cells with sera from rabbits immunized with purified SDS-denatured T Ag, but not with sera from hamsters bearing SV40-induced tumours. T-Ag-related antigens could be detected with both types of antisera by applying a more sensitive 125I-protein A assay. The T-Ag specificity of the binding of hamster SV40 tumour sera was demonstrated be a 125I-IgG-blocking assay in which preincubation of formaldehyde-fixed SV40-transformed cells with rabbiet anti-SDS T-Ag serum inhibited the binding of hamster SV40 tumour serum by about 70%. The localization of T-Ag-related antigens on the outside of plasma membranes of formaldehyde-fixed cells was shown by an anti-SDS-T-Ag serum-specific binding of fluorescein isothiocyanate-labelled Staphylococcus aureus to the cell surface. Out results are consistent with the hypothesis that SV40 T-Ag-related antigens are involved in the formation of TSTA. PMID- 6270238 TI - Intermediate size papovavirus particles in pregnancy urine. AB - By electron microscopy of negatively stained urinary sediments, papovavirus particles of size intermediate between papillomavirus and polymavirus have been detected in the urine of a pregnant woman. The excretion of size variants of these viruses and the absence of detectable papillomas in any of the women who were found to excrete only papillomavirus represent new findings. PMID- 6270239 TI - Herpes simplex virus infection in human monocyte cultures: dose-dependent inhibition of monocyte differentiation resulting in abortive infection. AB - Monocyte-enriched cultures of human blood leukocytes were exposed to herpes simplex virus (HSV) at different multiplicities of infection (m.o.i., from about 10 to 0.001 p.f.u./cell). Highest maximum progeny virus titres were invariably obtained with low initial m.o.i., i.e. those between 0.01 and 0.0001 p.f.u./cell, while little if any infectious progeny was produced in cultures inoculated with the highest virus concentrations. By the time of maximum virus production, i.e. 5 to 7 days after inoculation, monocytes in the uninfected cultures had mostly differentiated to macrophages. This differentiation was partially inhibited in cultures initially exposed to the higher concentrations of HSV. Synthesis of HSV antigens was detected by indirect immunofluorescence both in the high m.o.i. cultures and in the productively infected cultures. By this criterion, a maximum of 10 to 15% of all adherent cells became infected in both culture types. It is suggested that the higher doses of HSV, by inhibiting cellular maturation, also prevent the subsequent completion of its own infectious cycle. PMID- 6270240 TI - Lack of correlation between pp60src kinase activity and transformation parameters in cells infected with temperature-conditional mutants of Rous sarcoma virus. AB - The pp60src kinase activity of cells infected with temperature-conditional mutants of Rous sarcoma virus (RSV), which induce only a partial transformation, was compared to the pattern of transformation parameters induced by these mutants. The tsGI251-infected cells were thermosensitive for hexose transport, fibrinolysis, morphological alterations and pp60src kinase activity, but cold sensitive for loss of growth control. The tsGI253-infected cells were similar to tsGI251-infected cells except that they were only slightly cold-sensitive for loss of growth control and had a very low pp60src kinase activity which was temperature-insensitive. Thus, the pp60src kinase activity measured in vitro with IgG as a substrate did not correlate in a consistent way with any of the measured parameters of transformation. PMID- 6270241 TI - Effect of novobiocin and other DNA gyrase inhibitors on virus replication and DNA synthesis in herpes simplex virus type 1-infected BHK cells. AB - The four known inhibitors of the bacterial DNA gyrase (nalidixic acid, oxolinic acid, novobiocin and coumermycin A) were investigated with respect to their effect on the growth of uninfected BHK cells and the yield of virus from herpes simplex virus type 1 (HSV-1)-infected BHK cells. High concentrations of nalidixic acid and oxolinic acid (about 10 mM) were needed for 50% inhibition of cellular and viral multiplication with less than fourfold preferential inhibition of virus over cell growth. Novobiocin and coumermycin were effective at lower molar concentrations and the amount needed for 50% inhibition was 10-fold higher for cell growth than for virus yield. At 5 x 10(-4) M, novobiocin inhibited DNA synthesis in uninfected cells to approx. 20% of non-treated controls, while virus DNA in infected cells was almost completely inhibited (approx. 1% of controls). Residual cellular DNA synthesis in infected cells was rather insensitive (approx. 90% of controls) to this concentration of novobiocin. PMID- 6270242 TI - High yield growth and purification of human parainfluenza type 3 virus and initial analysis of viral structural proteins. AB - Structural proteins from a large-plaque variant (LPV) of human parainfluenza type 3 virus were analysed by electrophoresis on Laemmli-type polyacrylamide gels. High virus concentrations were obtained by growth in BS-C-1 cells cultivated on microcarrier beads. Purification of the virus in composite equilibrium gradients of potassium tartrate:glycerol resulted in 25% recovery of input infectivity and a preparation containing less than 0.08% of input host cell protein and RNA. Parainfluenza type 3 virus equilibrated at a density of 1.20 g/ml in these gradients. Analysis by polyacrylamide gel electrophoresis of 3H-glucosamine labelled virus taken from peak gradient fractions revealed 8 or 9 major virion peptides, ranging in mol. wt. from 17 x 10(3) to 125 x 10(3) (17K to 125K), two of which were glycoproteins. The sum of the estimated mol. wt. of these peptides, 501.5K to 570.5K, does not exceed the estimated genomic potential of other paramyxoviruses. PMID- 6270243 TI - Action of nucleotide derivatives on translation in encephalomyocarditis virus infected mouse cells. AB - The infection of animal cells by encephalomyocarditis (EMC) virus lead to a drastic change in membrane permeability towards low mol. wt. compounds. Addition of the nucleotide analogue GppCH2p to the culture medium resulted in a specific inhibition of protein synthesis in EMC virus-infected 3T6 cells. This inhibition was not observed when GTP or ATP were present nor in control mock-infected 3T6 cells. The induction of membrane leakiness after viral infection was not specific for 3T6 cells, as it was also detected in mouse L cells, hamster BHK-21 cells and monkey CV1 cells. The inhibitory action produced by GppCH2p in virus-infected cells was fully reversed upon addition of fresh medium. Moreover, analysis of the proteins synthesized after medium replacement showed a preferential synthesis of cellular proteins. The presence of zinc ions resulted in an inhibition of the cleavage of large viral polypeptide precursors to mature viral proteins. Under these conditions, membrane leakiness as measured by GppCH2p, was not observed. However, this seems to be an effect of zinc ions themselves on the membrane, because inhibition of mature protein formation by other means, such as the presence of amino acid analogues, did not prevent inhibition of translation by GppCH2p in virus-infected cells. Addition of the cap analogues 7mGppp and 2'-O' mGppp, resulted in specific stimulation of viral protein synthesis in EMC virus infected 3T6 cells. On the other hand, the presence of 7mGp had no effect on translation. We propose that a specific capping of viral mRNA takes place in the presence of these compounds, and leads to increased stability and greater efficiency in the translation of viral mRNA. PMID- 6270244 TI - Comparative mitogenic effects of herpes simplex virus and mycoplasma on murine lymphocytes. AB - Previous work indicated that herpes simplex virus type 1 (HSV-1) is a mitogen for mouse spleen cultures, as monitored by uptake of 3H-thymidine. We observed variable responses of mouse spleen cultures to different viral preparations. The variable responses, which did not follow normal dose-response relationships, were not due to HSV-1 strain differences, altered response kinetics or the presence or absence of defective viral particles, but to mycoplasma contamination of viral stocks. Mycoplasma-free (MF) HSV-1 stocks were prepared by transfection of MF NHF cells with HSV-1 DNA. MF HSV-1 infection of spleen cultures resulted in a five- to sixfold stimulation of DNA synthesis and stimulation of a polyclonal antibody response. Heat treatment (56 degrees C for 1 h) and antibiotics were used to distinguish mycoplasma and HSV-1-induced spleen culture mitogenic responses. The mycoplasma-induced mitogenic activity was found to be heat labile and sensitive to gentamicin and chloramphenicol. In contrast, the HSV-1 induced response was not affected by gentamicin or chloramphenicol and heat treatment resulted in only a 50% loss of activity. PMID- 6270245 TI - The detection of Epstein-Barr virus receptors utilizing radiolabelled virus. AB - Epstein-Barr virus (EBV) was labelled with 3H-thymidine and purified about 1000 fold from the culture medium by ultracentrifugation on 5 to 30% dextran gradients. The presence of the virus was monitored by radioactivity and Epstein Barr virus-determined nuclear antigen (EBNA) induction in sensitive indicator cells (Ramos). Peaks for both activities occurred in the 17 to 18% dextran fractions. Unlabelled virus recovered in the peak fraction was labelled with 125I. Both thymidine and 125I-labelled purified virus bound quantitatively to receptor-positive Burkitt lymphoma-derived cell lines but not to EBV-receptor negative T-lymphocyte-derived cell lines. Thymidine-labelled virus that was allowed to bind to Raji cells was present in the interior of briefly trypsinized cells after 3 h incubation at 37 degrees C. The results provide a convenient method for detecting the EBV receptor by radioactively labelled virus. PMID- 6270246 TI - Temperature-sensitive mutants in two distinct complementation groups of herpes simplex virus type 1 specify thermolabile DNA polymerase. PMID- 6270247 TI - Morphological components of herpesvirus. III. Localization of herpes simplex virus type 1 nucleocapsid polypeptides by immune electron microscopy. AB - Herpes simplex virus type 1 (HSV-1) nucleocapsids were observed in the electron microscope after their reaction with IgG's purified from the sera of rabbits immunized with the individual nucleocapsid polypeptides. The combining sites of NC1, the major capsid protein (mol. wt. 154K), were distributed over the entire capsid surface. This result provides further evidence that NC1 represents the major hexamer constituent. NC2 (mol. wt. 50K) was less widely distributed and appeared to be located at capsid vertices; that antigen may be a constituent of the pentamers or of peripentameric hexamers. One or both of NC3 and NC4 (mol. wt. 40K and 38K) were also located all over the capsid, possibly at positions interior to those of NC1. One or both may represent the intercapsomeric fibrils, hexamer-associated protein or material associated with the pericore. The locations of the other nucleocapsid polypeptides could not be determined. PMID- 6270248 TI - Genetic variation of neurotropic and non-neurotropic murine coronaviruses. AB - The murine coronavirus strains MHV JMH, MHV 1, MHV 2, MHV 3 and MHV A 59 were tested for their neurovirulence in weanling rats. The strain JHM was found to be highly neurovirulent for weanling rats, whereas the other strains were not, or only slightly, neurovirulent. MHV 1 caused no lesions in weanling rats. The other strains (MHV 2, MHV 3 and MVH A59) induced predominantly subclinical infections in weanling rats as demonstrated by an increase of antibodies and inflammatory lesions in the liver. Analysis of these strains by cross-neutralization revealed variable degrees of antigenic relationship between these viruses which were not related to their neurovirulence. However, when these strains were compared by analysing the T1-RNase-resistant oligonucleotides of virion RNA, the highly neurovirulent strain JHM was found to differ significantly in its nucleotide sequence from the other less-neurovirulent strains. PMID- 6270249 TI - Effect of mutation on the virulence in mice of a strain of foot-and-mouth disease virus. AB - Twenty-eight mutations, representing mutation in five different polypeptide coding regions of the foot-and-mouth disease genome, were examined for their effect on the virulence of the virus for suckling mice. Five types of mutation were examined: temperature-sensitive (ts), electrophoretic (e), co-variant temperature-sensitive and electrophoretic (ts/e), guanidine-resistant (gs+) and putative co-variant guanidine-resistant and electrophoretic (gs+/e). All the ts mutations and three out of the 11 non-ts mutations produced some reductions in virulence. In the majority of cases this reduction in virulence was shown to co vary with the mutation. No correlation was observed between the site of a mutation or its 'cut-off' temperature and the extent of the reduction in virulence. Studies of the growth in vivo of a small selection of ts mutants suggested that for most mutants their reduced virulence was a trivial effect of their slow growth rate. With one exception they all eventually grew to parental virus levels, the resulting virus being temperature-sensitive and the disease indistinguishable from that caused by the parental virus. The one exception was an avirulent ts mutant which only grew to one-thousandth the titre of the parent virus. This mutant did not cause disease and was therefore considered to be the only avirulent mutant. Its mutation was in the coat protein-coding region of the genome, probably the region coding for VP3. PMID- 6270250 TI - Cellular synthesis and modification of murine hepatitis virus polypeptides. AB - Mouse L fibroblasts infected with mouse hepatitis virus, MHV3, and radiolabelled with 35S-methionine, contained, in addition to the three major structural polypeptides, p24, p56 and p180, two additional ones, p22 and p50. Of these total five polypeptides, only three, p22, p56 and p180, were labelled in infected cells during a 2 min 35S-methionine pulse and are, therefore, presumed to be immediate translation products. Pulse-chases and chymotryptic peptide mapping experiments showed apparent precursor-product relationships between p56 and p50 and between p22 and p24. Protein synthesis in infected cells was synchronized at the initiation stage by pre-exposure to hypertonic medium. Using a 0.5 min pulse-10 min chase sequence, to limit incorporation of 35S-methionine to stretches of approx. 100 amino acids adjacent to translational initiation sites, it was found that all three polypeptides, p22, and p56 and p180 contained radiolabel. It is thus apparent that translation of the three major structural proteins (or precursors) is initiated independently rather than at a single site as in the cases of other positive-strand RNA viruses such as polio or Semliki Forest virus. PMID- 6270251 TI - Isolation and characterization of feline rotavirus. AB - Feline rotavirus was detected by electron microscopy in the faecal samples of a cat, and was propagated in an established cell line of foetal rhesus monkey kidney, MA104, cell cultures. Morphologically, feline rotavirus was indistinguishable from known rotaviruses. Complete particles showed a characteristic "spoke-like' arrangement of inner capsomers surrounded by an outer layer. Intracytoplasmic inclusion bodies in different sizes and shapes were produced in infected MA104 cells. Reproducible clear-cut plaques were produced by feline rotavirus in MA104 cells under the overlay of carboxymethylcellulose in the presence of trypsin. Feline rotavirus was distinct from human, canine, bovine, porcine and simian rotaviruses by the plaque reduction neutralization test. Feline rotavirus, like canine and simian rotaviruses, was found to be less dependent upon trypsin than human, bovine, porcine, chicken and turkey rotaviruses. A seroepidemiological survey (September 1979 to August 1980) showed that 20 out of 61 (32.8%) randomly sampled hospitalized cats at the Cornell Veterinary Teaching Hospital in Ithaca, New York had antibody titres against feline rotavirus. Oral inoculation of cats with feline rotavirus did not produce any clinical disease, but most cats did mount an immune response to the virus following inoculation. PMID- 6270252 TI - Isolation and characterization of a temperature-sensitive uncoating mutant of pseudorabies virus. AB - During the course of characterizing a series of temperature-sensitive mutants of pseudorabies virus, we found one (designated tsL) that did not produce cytopathic changes in rabbit kidney cells at the non-permissive temperature (41 degree C). Although the mutant adsorbed to and penetrated the cells in a normal fashion, virus RNA was not synthesized at 41 degree C in the infected cells. However, if the cells were first incubated at the permissive temperature (32 degree C), virus RNA synthesis occurred at the non-permissive temperature. This occurred even if, during the incubation period at 32 degree C, the expression of viral functions was prevented by treatment with an inhibitor of protein synthesis. The DNA in tsL virions did not appear in the cell nucleus at 41 degree C, and full, non enveloped nucleocapsids could be recovered from the cytoplasm of tsL virus infected cells. These results show that the nucleocapsids of tsL remain intact at the non-permissive temperature and that tsL is an uncoating mutant. PMID- 6270253 TI - The nature of the DNA associated with incomplete particles of adenovirus type 2. AB - The nature of the DNA in incomplete particles (IP) synthesized by adenovirus type 2 and the ts4 mutant which accumulates such particles were analysed by agarose gel electrophoresis, restriction endonuclease cleavage and blot hybridization techniques. IP DNA consisted of a heterogeneous population of subgenomic-size DNA (IPSD1) and smaller molecules ranging from about 1000 base pairs to 200 base pairs (IPSD2). IPSD1 from ts4 was more heterogeneous than that from wild-type (wt), but both contained sequences from all parts of the viral genome. IPSD2 contained heterogeneous cellular sequences and viral sequences from the left 4.4% of the genome. An endonuclease activity associated with IP and virions was capable of digesting viral or cellular DNA to IPSD2-like fragments suggesting a possible origin for these molecules. PMID- 6270254 TI - Establishment and propagation of a bovine leukaemia virus-producing cell line derived from the leukocyte of a leukaemic cow. AB - A cell line (LB59Ly) derived from the leukocytes of a leukaemic cow was established as a monolayer, which spontaneously released large amounts of a retrovirus. This virus was found to be indistinguishable from the bovine leukaemia virus (BLV) produced by the commonly used high-producing heterologous cell line (FLK-BLV). Like the latter, its reverse transcriptase activity was greater in the presence of Mg2+ cations than in the presence of Mn2+ cations; its polyacrylamide gel electrophoresis pattern showed the presence of gp51, p24, p15, p12 and p10, and the antigenicity of the two major proteins completely cross reacted with those of BLV from FLK-BLV cells. The virus was infectious and induced early and late polykaryocytosis, the specificity of which was demonstrated by use of specific anti-BLV sera. PMID- 6270255 TI - The relationship between genomic RNAs of polycythaemic forms of spleen focus forming Friend virus and its helper virus. AB - We have studied the relationship between Friend spleen focus-forming virus (SFFV) and its helper lymphoid leukaemia virus (LLV) by comparing RNase T1 fingerprints of genomic RNAs. Our data indicate that about 70% of the SFFV sequence is a perfect copy of parts of the helper genome. We conclude that our SFFV and LLV isolates have co-evolved very closely and that SFFV-specific sequences are not identical in different Friend virus isolates. PMID- 6270256 TI - The spontaneous and induced synthesis of Epstein-Barr virus antigens in Raji cells immobilized on surface coated with anti-lymphocyte globulin. AB - Immobilization of Raji cells on surface coated with anti-lymphocyte globulin (ALG) at low cell densities lead to the synthesis of Epstein-Barr virus (EBV) early antigen (EA) in up to 5% of the cells. At higher cell densities the percentage of antigen-positive cells decreased and at confluency no antigen synthesis was observed. Addition of iododeoxyuridine (IdUrd) to low density cultures increased the expression of EA to 20%, whereas in confluent cultures the cells could not be induced to synthesize EA. Treatment of cells in suspension with ALG failed to induced EA synthesis and did not potentiate the effect of IdUrd. Immobilized Raji cells proved to be suitable targets for superinfection with EBV derived from P3HR1 cultures. PMID- 6270257 TI - Varicella-zoster virus infection of diploid and chemically transformed guinea-pig embryo cells: factors influencing virus replication. AB - Factors influencing the replication of varicella-zoster virus (VZV) in guinea-pig embryo cells were evaluated using both diploid cells (GPEC) and a chemically transformed cell line (GPT). Wild-type and vaccine strains of VZV were successfully isolated and serially propagated in GPEC prepared from early gestation whole embryos (less than 2 cm in length). Low passage GPEC (less than or equal to 5 subcultivations) were more susceptible to VZV infection than high passage GPEC (greater than 5 subcultivations), and guinea-pig cells were consistently less permissive than human diploid cells. Cell-free virus was produced from VZV-infected GPEC cultures by sonication and peak yields of 10(3) p.f.u./ml were obtained. In addition, we report the isolation and propagation of VZV, as well as production of cell-free virus, in GPT. Both GPEC and GPT cells were less susceptible to VZV infection than human cells. However, viral replication was enhanced by incubation of VZV-infected GPT cultures at 32 degree C rather than 36 degree C. PMID- 6270258 TI - Transcription and translation of the herpes simplex virus type 1 thymidine kinase gene after microinjection into Xenopus laevis oocytes. AB - The hybrid plasmid pTK1 consists of the herpes simplex virus type 1 (HSV-1) BamHI p fragment, which contains the thymidine kinase (TK) gene, inserted into the vector pAT 153. When pTK1 DNA was microinjected into nuclei of Xenopus laevis oocytes, functional HSV-1-specific TK was produced, showing that transcription and translation of the gene occurred. Investigation of pTK1-specific RNA by "Southern' blot hybridization revealed that all regions of the hybrid plasmid were transcribed by RNA polymerase II, but sequences present in TK mRNA were most highly represented in stable transcripts. PMID- 6270260 TI - Comparison of the immediate early polypeptides of human cytomegalovirus isolates. PMID- 6270259 TI - Infection of mouse ectoplacental cone cells with murine cytomegalovirus. AB - In order to understand the mechanism of congenital cytomegalovirus (CMV) infection, we studied the effect of murine cytomegalovirus (MCMV) on murine ectoplacental cone (EPC) cells in vitro. Cytopathic effects (c.p.e.) were seen in many MCMV-infected EPC cell cultures 5 to 7 days after exposure to MCMV. The infected cells showed intranuclear inclusions characteristic of CMV infection when stained with May-Grunwald-Giemsa. Culture fluids collected from MCMV infected EPC cells after 4 or more days of culture caused c.p.e. when co cultivated with mouse embryo fibroblasts (MEF). Employment of the anti-complement immunofluorescence (ACIF) test detected MCMV-specific antigens, in situ hybridization localized MCMV DNA and electron microscopy detected the presence of the viral particles in the MCMV-infected EPC cells. Thus, exposure of EPC cells to MCMV in vitro resulted in a productive infection. PMID- 6270261 TI - A hybridoma cell line secreting antibody to poliovirus type 3 D-antigen: detection in virus harvest of two D-antigen populations. AB - A mouse hybridoma cell line (Mo56) secreting IgG antibody to poliovirus type 3 D antigen was obtained by fusion of a mouse myeloma cell line with spleen cells from mice immunized with Saukett virus. The monoclonal antibody was specific for Saukett virus strains in virus-neutralization and single-radial-diffusion tests. In immunoprecipitation tests the monoclonal antibody reacted with intact infectious virus particles (155S) and with a previously undescribed 70S poliovirus particle with D-antigenic reactivity. PMID- 6270262 TI - Host restriction property of a vesicular stomatitis virus mutant isolated from carrier cultures. AB - Carrier cultures of L cells infected with wild-type vesicular stomatitis virus (VSV0) were initiated without the use of defective-interfering particles or homologous interferon. The cloned viruses recovered from such carrier cultures after passage 21 were characterized as temperature-sensitive. Furthermore, these clones of the mutant showed restricted replication at permissive temperature in HEp-2 cell cultures as compared to the wild-type VSV0. This restrictive replication of the mutant in HEp-2 cells was not due to a defect in the expression of virion-associated primary transcriptase activity in vivo, but due to the marked reduction in virus-specific characterized may govern the synthesis of mutant virus-specific amplified RNA in HEp-2 cells. PMID- 6270263 TI - Modification and exploitation of a poliovirus-induced membrane complex by superinfecting ME virus. AB - The replication of Mouse Elberfeld (ME) virus was accelerated when HEp-2 cells were mixedly infected with poliovirus in the presence of guanidine. The latent period of the replication of ME virus was shortened by 3 h when cells were preinfected for at least 2 h with poliovirus and inhibited by guanidine. Simultaneous infection with poliovirus and ME virus resulted in a shortening by 1 h of the latent period of ME virus replication. The accelerated replication of ME virus was shown to be due to modification and exploitation of a membrane complex induced by poliovirus in the presence of guanidine; on superinfection ME virus successively modified this poliovirus-induced complex of 470S ("light' complex) into a "heavy' complex of 700S specific for ME virus. PMID- 6270264 TI - Effect of enzymes on the attachment of influenza and encephalomyocarditis viruses to erythrocytes. AB - Encephalomyocarditis (EMC) and influenza viruses attach to human erythrocytes causing haemagglutination of the cells. Sialoglycoproteins, containing predominantly glycophorin A, from these cells behave as soluble virus receptors and inhibit haemagglutination by both viruses. Removal of 43% of the sialic acid from erythrocytes with neuraminidase prevented their haemagglutination by EMC virus loss of 40% of glycophorin sialic acid destroyed its inhibitory properties against this virus. However, about 80% of the sialic acid had to be removed from erythrocytes or from glycophorin to achieve the same results for influenza virus. Trypsin treatment of erythrocytes or glycophorin had little effect on haemagglutination or inhibition involving either virus, although the glycopeptides released contain up to 70% of the total sialic acid, and despite the fact that glycophorin was drastically reduced in size as shown by SDS- polyacrylamide gel electrophoresis. It is concluded that not all of the sialic acid present in erythrocyte sialoglycoprotein receptors is involved in attachment of EMC or influenza viruses and that the attachment sites on erythrocytes for these viruses are not identical. PMID- 6270266 TI - Nucleotide sequences of the joint between the L and S segments of herpes simplex virus types 1 and 2. AB - The a sequence of herpes simplex virus (HSV) is present as a direct repeat at the genomic termini and also in inverted orientation at the joint between the L and S segments. DNA sequences have been determined for the joint regions of the genomes of HSV-1 and HSV-2, and relative to these sequences the genomic termini are in both cases located close to a short direct repeat of 17 to 21 base pairs (bp) at the b-a and a-c junctions. The HSV-1 joint region contains three separate tandem direct reiterations of short sequences, (12, 16 and 17 bp in strain 17) and we conclude that size heterogeneity in the a and c sequences is due to variable copy numbers of these repeated units. It is likely that a considerable part of the HSV 1 joint region does not code for polypeptide. PMID- 6270265 TI - Conversion of herpetic lesions to malignancy by ultraviolet exposure and promoter application. AB - Many lines of evidence exist associating herpes simplex virus (HSV) with the development of carcinoma, but much of this evidence is anecdotal or associative in nature and does not prove a cause and effect. The purpose of this research was to investigate the oncogenic potential of HSV type 2 (HSV-2) in vivo. A mouse model for lip carcinogenesis was designed to combine HSV-2 infection, u.v. exposure and tetradecanoyl-phorbol-acetate (TPA) application. HSV-2 inoculation on to abraded mouse lips was capable of causing vesicular ulcerative lesions which healed completely after 10 to 14 days. Ultraviolet irradiation of the HSV lesion site daily for 6 min at 42 ergs/mm2/s on days 3 to 6 post-infection caused hyperkeratosis, acanthosis and dysplasia to develop in several lips; while the same u.v. exposure by itself failed to alter the histologic appearance. The addition to repeated TPA application to the HSV+ u.v. regimen promoted tumour emergence. Thirty-two of 156 Balb/c mice developed tumours. Although the majority were papillomas, six were squamous cell carcinomas. These tumour-bearing mice had increased HSV-specific antibody titres. HSV antigens were shown to be present in outgrowths from explanted tumours as well as in tumour biopsies by immunoperoxidase staining with HSV-specific antisera. It is suggested that the in vivo u.v.-irradiated HSV acted as the inducer and TPA as the promoter, analogous to the classical two-stage carcinogenesis model. PMID- 6270267 TI - Epstein-Barr virus-induced proteins. IV. Characterization of an EBV-associated phosphopolypeptide. AB - Cells of the Epstein--Barr virus (EBV) non-producing lines NC37 and Raji were induced by the tumour promoter TPA and were labelled with 32P. The analysis by immunoprecipitation with human VCA+EA+ sera revealed that a major phosphopolypeptide with a mol. wt. of 58 000 was specifically precipitated. In addition, a phosphopolypeptide of the same size was the dominant phosphopolypeptide detected in EBV-producing B95-8 and P3HR-1 cells as well as in P3HR-1-superinfected NC37 cells. This phosphopolypeptide seemed to be EBV specific, because it was not detectable in TPA-treated, but EBV genome-negative, Ramos and BJAB cells. Synthesis of this phosphopolypeptide was inhibited in TPA treated NC37 cells by treatment with Ara-C. VCA+EA+ sera were found to be more immunoreactive with this phosphopolypeptide than VCA+EA- or EBNA+ sera. Based on these results this phosphopolypeptide may have some immunological relatedness to the EBV-specific early antigen (EA) complex defined by immunofluorescence. PMID- 6270268 TI - Cytopathic effects in mouse neuroblastoma cells during a non-permissive infection with a mutant of vesicular stomatitis virus. AB - Morphological changes were extensive following infection of murine neuroblastoma N-18 cells with a temperature-sensitive (ts) mutant of vesicular stomatitis virus (VSV), G31 (complementation group III), and incubation at 39 degrees C, a non permissive condition for virion maturation. Incubation for 24 h after infection resulted in extensive morphological degeneration of mitochondria with over 80% from that in uninfected cells. Janus green B supravital staining, was reduced by 81% from that in uninfected cells. Cellular ATP levels were reduced by 50% 12 h after infection. Mitochondrial degeneration still occurred in infected cells after the inactivation of lysosomes with chloroquine. Extensive cell fusion and cytoplasmic vacuole formation also occurred during the non-permissive infection with ts G31. Loss of plasma membrane integrity was not the cause of vacuole formation since 90% of the cells were able to exclude trypan blue 24 h after infection, nor were the vacuoles the result of inactivation of the mitochondria since cyanide-poisoned cells did not form vacuoles. The cytopathic alterations observed in N-18 cells during the non-permissive infection of N-18 cells with ts G31 did not occur during the non-permissive infection of N-18 cells with ts G11 (I), ts G41 (IV), or u.v.-inactivated ts G31. However, the non-permissive infection with ts O45 (V) led to mitochondrial degeneration and cytoplasmic vacuole formation, but no cell fusion occurred. These results are discussed in light of the ultrastructural features previously observed in the central nervous system of mice infected with ts G31 and cells in culture infected with wild-type VSV. PMID- 6270269 TI - Kinetics of inhibition of papovavirus DNA synthesis by superinfection with adenovirus 2 and non-defective adenovirus 2-simian virus 40 hybrid viruses. AB - Simian Virus 40 (SV40) DNA synthesis is inhibited in monkey cells by superinfection with adenovirus 2 (Ad2) and various non-defective Ad2-SV40 hybrid viruses. Similarly, BKV (a human papovavirus) DNA synthesis is inhibited in human cells by superinfection with Ad2. Kinetic studies indicate that inhibition begins during the early phase of the Ad2 lytic cycle. Superinfection with Ad2 does not significantly alter the formation of SV40 T antigen. Superinfection with Ad2 late in SV40 lytic cycle is less efficient in the inhibition of SV40 DNA synthesis, and the onset of Ad2 DNA synthesis is delayed, compared to superinfection early in the SV40 lytic cycle. These findings suggest that the Ad2 and SV40 genomes may compete to bind an early AD2 protein which is essential for Ad2 replication, but which blocks SV40 replication. PMID- 6270270 TI - Comparison of T antigen-associated host phosphoproteins from SV40-infected and transformed cells of different species. AB - Simian virus 40 (SV40)-infected and -transformed cell contain, in addition to the virus-coded tumour antigens, one or more 48K to 56K host tumour antigens. At least part of this class of host proteins exists as a fast-sedimenting complex with the SV40 large T antigen. The host proteins associated with the large T antigen in SV40-transformed monkey, mouse and human cells and SV40-infected monkey cells were compared by two-dimensional gel electrophoresis and V8 partial proteolysis peptide mapping. Although these proteins differed slightly in apparent mol. wt. and peptide pattern, they migrated identically in isoelectric focusing gels. These results suggest that the cellular proteins associated with large T antigen in different hosts are very closely related to each other. Despite their similarities, the 55K proteins from different host cells form complexes of different stabilities with large T antigen, as judged by spontaneous dissociation of the complexes during storage, and the fractions of the 55K cellular protein and large T antigen found free and in the complexed form in each different host cell. PMID- 6270271 TI - Terminal structure of reovirus RNAs. AB - S1 nuclease analysis and 3' terminal sequencing of the reovirus genome double stranded RNAs and in vitro produced viral mRNAs has been used to establish the viral mRNA is a full length copy of the genome template. Sequence determination at the 3' end of the genome minus strand has by transposition allowed determination of the 5' terminal sequences of the viral mRNAs. In all but one case an AUG codon which could function in initiation of protein synthesis has been found within the determined sequence. The 3' ends of the plus strands from all genome segments were found to have a common sequence. The implications of these results on the mechanism of virus replication are discussed. PMID- 6270272 TI - Isolation from chickens of a rotavirus lacking the rotavirus group antigen. AB - A virus, designated 132 virus, was isolated from the faeces of chickens in chick embryo liver cell cultures. The morphology and morphogenesis of 132 virus were indistinguishable from that of rotaviruses. The nucleic acid of 132 virus had the nuclease resistance of double-stranded RNA, and was separated by polyacrylamide gel electrophoresis into 11 segments with mol. wt. ranging from 2.07 x 10(6) to 0.20 x 10(6). SPF chickens were susceptible to oral infection with 132 virus, which replicated in the villous epithelial cells of the small intestine. 132 virus was therefore a rotavirus by morphological, biochemical and biological criteria. However, by immunofluorescence it was not possible to demonstrate an antigenic relationship between 132 virus and known avian and mammalian rotaviruses, indicating that 132 virus does not possess the group antigen shared by all previously characterized rotaviruses. This finding has implications for the diagnosis of rotavirus infections by serological tests. PMID- 6270273 TI - Proteolytic cleavage of VP1 in 'A' particles of coxsackievirus B3 does not appear to mediate virus uncoating by HeLa cells. AB - 'A' particle of Coxsackievirus B3 were generated from native virus by heating and purified by sucrose gradient centrifugation. These particles were found to be similar to 'A' particles formed by elution from cellular receptors of HeLa cells. Electrophoretic analysis of [35S]methionine-labelled 'A' particles revealed that treatment of the particles with chymotrypsin resulted in the cleavage of VP1 and the formation of a cleavage product which migrated between VP2 and VP3. Analysis of the protease-treated material on sucrose gradients revealed a ribonuclease sensitive particle which sedimented more slowly than an 'A' particle. This particle apparently degraded to release the viral RNA, thereby providing an in vitro model for protease-mediated uncoating of 'A' particles. The subviral particles of Coxsackievirus B3 were found to be immunoprecipitable with heterotypic Coxsackievirus group B antisera, thereby providing a method for the recovery of products produced in the cell early in infection. Infected cells which had been treated to remove unreacted virus were disrupted, an the lysates were reacted with heterotypic antisera. Analysis of the precipitated material revealed that no cleavage products were formed and no polypeptides were lost. Therefore, it appears that proteolysis is not involved in the uncoating of Coxsackievirus B3 in infected cells. PMID- 6270274 TI - An intracellular interaction between a temperature-sensitive mutant and the original wild-type HVJ (sendai virus) is responsible for the establishment and maintenance of HVJ persistent infection. AB - In order to understand the selective survival of temperature-sensitive (ts) mutants in persistent infection by HVJ (Sendai virus), an intracellular interaction between a ts clone (HVJ cl.14) isolated from HVJ carrier G2 cells and the original wild-type virus (HVJo) was studied. HVJ cl.14 differed from HVJo mainly in its ts property at 39 degrees C, weak cytopathogenicity and faster electrophoretic mobility of P protein (P77K), but showed similar trypsin activated growth to that of HVJo. When LLCMK2 cells were simultaneously infected with HVJo and HVJ cl.14 at 32 degrees C, synthesis of HVJo-derived P protein (P79K) was inhibited with concomitant reduction of cytopathic effect (c.p.e.) and more dominant growth of HVJ cl.14 was observed. For the analysis of progeny viruses in these mixed infections, another mutant of HVJo designated HVJe which formed plaques activated only by elastase was isolated and employed instead of HVJo. At 39 degrees C, HVJ cl.14 was rescued by coinfected HVJe at about 900- to 13 000-fold over single infection. This recovery was also shown by sequential synthesis of HVJ cl.14-derived P protein (P77K) following the earlier synthesis of HVJo-derived P polypeptide (P79K) in the mixed infection at 39 degrees C. However, the u.v. inactivation of HVJe or HVJ cl.14 resulted in a loss of their activity on rescue or on c.p.e. reduction, suggesting the necessity of protein synthesis by opposite viruses for these interactions. The mechanisms involved in the predominant growth of the ts mutant and concomitant reduction of c.p.e. seemed to provide a general explanation for the preferable persistence of the ts mutant in the HVJ carrier cells. PMID- 6270275 TI - Temperature-sensitive HVJ (sendai virus) with altered P polypeptide derived from persistently infected cell lines. AB - HVJ isolated from culture fluids of G2, THEL and GM2 cells persistently infected with HVJ (G2-HVJ, THEL-HVJ and GM2-HVJ) were characterized in comparison with wild-type HVJ (HVJo). Viral structural proteins were analysed by 10% SDS polyacrylamide gel electrophoresis and its was found that only the P polypeptides of all the HVJ clones isolated from G2-HVJ cells had a smaller size mol. wt. of 77 000 (77K), than that of HVJo with a mol. wt. of 79 000 (79K). One of six clones from THEL-HVJ cells and one of ten clones from GM2-HVJ cells exhibited the same migration pattern of P polypeptide as that of the clones from G2-HVJ cells. However, the other structural proteins were not different from those of the wild type virions. All the clones from these carrier cultures were temperature sensitive and were blocked in early step(s) required for RNA synthesis. These results indicate that some mutations(s) associated with P polypeptides could occur during the course of HVJ persistent infection in cell cultures. PMID- 6270276 TI - Correlation of in vitro properties and tumourigenicity of hamster kidney cells transformed by BK virus. AB - Hamster kidney cells transformed by BK virus (HKBK cells) were studied at low passage (about 30 subcultures) after transformation and at high passage (about 130 subcultures) after transforming. Several in vitro properties (rescue of virus, presence of T antigen, colony formation on plastic, in soft agar, on monolayers of normal cells, and serum dependency of growth) and tumourigenicity of these cells for newborn and adult hamsters were compared. HKBK cells at low passage showed high levels of T antigen, the growth properties of normal cells and low tumour-producing ability, while HKBK cells at high passage showed low levels of T antigen, the growth properties of transformed cells and high tumour producing ability. These results may indicate that cellular transformation by BK virus is initiated, but not maintained, by the expression of genes regulating T antigen, and that the maintenance of the transformed state is due to complicated cellular and viral gene interactions occurring during the course of cell life. PMID- 6270277 TI - Interactions between polymerized human albumin, hepatitis B surface antigen, and complement: II. Involvement of Clq in or near the hepatitis B surface antigen receptor for polyalbumin. AB - A species-specific receptor for polymerized human albumin (PHALB) has been reported on hepatitis B surface antigen (HBsAg)-carrying particles. Our previous observations that human Clq also binds PHALB in a species-restricted manner led us to investigate the possibility that HBsAg-associated Clq is involved in the PHALB receptor on HBsAg particles. The temperature, ionic strength, and pH requirements necessary for binding of PHALB to both Clq and HBsAg were compared and found to be similar. Normal human serum and purified Clq inhibited the PHALB HBsAg interaction; the inhibition was markedly reduced in heat-inactivated and Clq-depleted serum. Heat-denatured or reduced and alkylated Clq failed to inhibit the PHALB-HBsAg binding. Moreover, human Clq was found to be present in purified preparations of HBsAg and the quantity detected paralleled the degree of PHALB HBsAg binding. While anti-Clq inhibited the PHALB-HBsAg interaction, anti-Clr, - Cls, -C3, and -Ig were not inhibitory. Collagenase treatment of purified HBsAg reduced both PHALB-binding activity and the degree of HBsAg-associated Clq. These observations provide evidence that HBsAg-associated Clq is involved in or near the HBsAg-binding site for PHALB. PMID- 6270278 TI - Prevalence of antibody to human calicivirus in general population of northern Japan. AB - Serum specimens from children and adults living in Saporo, Japan, were tested for antibody against human calicivirus by immune electron microscopy (IEM), using virus-rich faecal extracts as the source of antigen. Of 83 serum specimens tested, 49 (59%) were positive for calicivirus antibody. Age-related prevalence of antibody to calicivirus was as follows: 23% (3/13) in the 0-5-month-old group, 30% (6/20) in the 6-23-month-old group, 65% (13/20) in the 2-5-year-old group, and 90% in school children (18/20) and adults (9/10). As for IEM antibody ratings scored from 0 to 4, almost all positive sera from older infants and preschool children scored 3 to 4. Antibody scores were rather more scattered in school children. The results indicated that caliciviral infection is prevalent in younger children in this part of Japan. PMID- 6270279 TI - Immunogenicity of A/USSR (H1N1) subunit vaccine in unprimed young adults. AB - A clinical and serological study was performed on 267 of 636 volunteers vaccinated against Argentine hemorrhagic fever with the XJCl3 attenuated strain of Junin virus seven to nine years earlier, in order to determine their long-term evolution. This study included a clinical examination, a chest roentgenogram, an electrocardiogram, and the following laboratory determinations: white and red cell count, number of platelets, hematocrit, hemoglobin, sedimentation rate (Katz index), urea, nitrogen, glucose concentration, cholesterol, GOT, GPT, gamma GT, alkaline phosphatase, cholinesterase, and total bilirubin. Neutralization reactions were performed to determine presistence of antibody levels. All clinical and laboratory findings were within normal limits, excluding a long-term pathology attributable to the virus. Of 165 tested sera, 153 (90.3%) had detectable levels of neutralizing antibodies, and the rest had no antibodies after this time. Although these people live in the endemic area, it is considered that only the 9% that had increased antibody levels had suffered a reinfection during the seven- to nine-year period, which acted as a booster. This figure aproximately coresponds to the subclinical infection value found in the region. In the rest, the persistence of antibodies is attributed to the immunization achieved with the vaccine employed. PMID- 6270280 TI - Propagation of hepatitis A virus in human embryo fibroblasts. AB - human diploid fibroblasts and human primary liver cell carcinoma cells (PLC/PRF/5) were infected with hepatitis A virus (HAV) adapted to growth in cell culture or derived directly from human stool. Viral antigen was expressed in PLC/PRF/5 cells 28 days after infection with cell culture-adapted HAV, and 50 days after infection with virus from human stool. In human fibroblasts the periods until first expression of viral antigen were 90 and 210 days, respectively. During further passages of HAV in fibroblasts the time of first appearance of antigen decreased to about 28 days. Biophysical properties of HAV extracted from infected fibroblasts were comparable to those of HAV derived directly from human stool. Immunofluorescence studies showed that the antigen was located exclusively within the cytoplasm of the infected fibroblasts. Kinetics of antigen production indicated that an equilibrium between virus multiplication and cell metabolism was reached in persistently infected fibroblasts. PMID- 6270281 TI - Effects of adenosine 3',5'-cyclic monophosphate on thermoregulation in both rats and rabbits. AB - The effects of intraventricular administration of dibutyryl adenosine 3',5' cyclic monophosphate (db cyclic AMP) on the thermoregulatory responses of unanesthetized rats and rabbits to different ambient temperatures (Ta) were assessed. Administration of db cyclic AMP (10-60 mM) produced dose-dependent hypothermia in both rats and rabbits at Ta 2-22 degrees C. The hypothermia in response to db cyclic AMP was due to decreased metabolic heat production and cutaneous vasodilatation. There was no change in respiratory evaporative heat loss. In contrast, in the heat (30-32 degrees C), db cyclic AMP administration produced dose-dependent hyperthermia in these animals. The hyperthermia was due to increased metabolism (due to muscular shivering) and decreased heat losses. The reduction in heat losses was shown by a decrease in both cutaneous circulation and respiratory evaporative heat loss. The data demonstrate that the thermoregulatory responses induced by central administration of db cyclic AMP are Ta-dependent. PMID- 6270282 TI - Studies on erythrocyte membranes of patients with Huntington's disease. AB - Several red cell membrane properties and activities of membrane-bound enzymes were investigated in blood samples of patients with Huntington's disease. (Na(+)+K(+)) ATPase activity and cell deformability appeared to be normal, in contradiction to preceding reports from other laboratories. With other techniques sensitive to relatively small changes in membrane structure, no abnormalities were found in Huntington's disease red cell membranes. These investigations do not support the concept that a generalised membrane abnormality is present in Huntington's disease. PMID- 6270283 TI - Viruses, Parkinsonism and Alzheimer's disease. PMID- 6270284 TI - Postsynaptic localization of alpha 2-adrenergic receptors in rat submandibular gland. AB - Norepinephrine is known to inhibit its own release from presynaptic nerve terminals through alpha 2-adrenergic receptors, which presumably have a presynaptic localization. alpha 2-Adrenergic receptors (as determined by [3H]clonidine binding) appear in rat submandibular gland membranes following reserpine treatment. These alpha 2 receptors seem to be localized postsynaptically, based on the following evidence. (1) Partial destruction of the presynaptic nerve terminals with 6-hydroxydopamine did not decrease the density of alpha 2-adrenergic receptors following subsequent reserpine administration. (2) Duct ligation, which results in atrophy of the gland, markedly decreased the density of the receptors following subsequent reserpine administration. (3) Surgical denervation resulted in the appearance of high levels of alpha 2 adrenergic receptors. (4) The changes in alpha 2 receptors paralleled the changes in postsynaptic beta-adrenergic receptor binding (as determined by [3H]dihydroalprenolol). While these results establish the existence of postsynaptic alpha 2-adrenergic receptors in an innervated tissue, the concomitant presence of a low density of presynaptic alpha 2 receptors has not been eliminated. PMID- 6270285 TI - Picrotoxinin and diazepam bind to two distinct proteins: further evidence that pentobarbital may act at the picrotoxinin site. AB - alpha-[3H]Dihydropicrotoxinin (DHP) and [3H]diazepam binding proteins were solubilized from rat brain membranes with 1% Lubrol-Px. Gel filtration of the Lubrol-solubilized fraction revealed that [3H]DHP and [3H]diazepam bind to two distinct peaks with apparent molecular weights of 185,000 and 61,000, respectively. The signal-to-noise ratio of [3H]DHP binding to 185,000-dalton fractions was improved significantly. [3H]DHP bound to the 185,000-dalton fraction with two binding constants. Muscimol and pentobarbital, while enhancing [3H]diazepam binding to membrane and crude Lubrol-solubilized fractions, failed to enhance [3H]diazepam binding to the 61,000-dalton fraction. Pentobarbital inhibited the binding of [3H]DHP to the 185,000-dalton fraction with an IC50 value of 60 +/- 12 micro M. The binding of [3H]DHP alos was inhibited by several depressant and convulsant drugs which affect gamma-aminobutyric acid (GABA) mediated transmission. These results provide strong evidence that picrotoxinin and diazepam bind to two distinct proteins and that pentobarbital may act at the picrotoxinin-sensitive site of the benzodiazepine . GABA receptor . ionophore complex. PMID- 6270286 TI - The action of nerve growth factor and dibutyryl adenosine cyclic 3':5' monophosphate on rat pheochromocytoma reveals distinct stages in the mechanisms underlying neurite outgrowth. AB - The clonal rat pheochromocytoma, PC12, responds to nerve growth factor (NGF) and dibutyryl adenosine cyclic 3':5'monophosphate (dbc AMP) by the elevation of cellular protein and RNA levels in all three parameters that are additive or greater than the sum of those caused by either agent alone, indicating that the mechanisms by which the two agents act to produce these changes are distinct. The concentration of dbcAMP required for half-maximal stimulation of these changes is also different for each, while NGF is active in all instances at 10(-11) M. PC12 cells initially generate neurites slowly in response to NGF and, at the same time, develop the capacity to regenerate neurites rapidly, a process termed priming. The cells, however, are not primed by dbcAMP nor does it influence the ability of NGF to prime them. Time lapse cinematography demonstrates that both NGF and dbcAMP each have unique effects on cellular morphology. The latter produces' rapid, unstable neurite initiation but does not promote sustained neurite extension. In contrast, NGF has immediate effects at the cell surface with no neurite initiation but later produces sustained neurite outgrowth. Utilizing dbcAMP, it is possible to dissect four morphological response of PC12 cells to NGF. Three of these may be mechanistically closely tied to occupancy of the NGF plasma membrane receptor. PMID- 6270287 TI - Uterine activity: a potential cause of false-positive Meckel's scans. PMID- 6270288 TI - Role of pyridoxal kinase in vitamin B6 uptake by Escherichia coli. AB - Escherichia coli KG980, a vitamin B6 auxotroph derived from wild strain K12, concentrated exogenous pyridoxal in an energy-dependent manner, and the effects of energy sources and inhibitors on pyridoxal uptake, compared with those on proline uptake indicated that the energy required was in the form of phosphate bonds and not of membrane potential. The vitamin taken up was primarily present as pyridoxal 5'-phosphate and pyridoxamine 5'-phosphate intracellularly, and energy depletion decreased the accumulation as the phosphorylated derivatives but not as unaltered pyridoxal itself. This finding suggested that the intracellular phosphorylation, which was known to require ATP, was essential for the concentrative uptake of the vitamin. The suggestion was confirmed by the following evidence. 1) Pyridoxal oxime inhibited pyridoxal uptake by decreasing the intracellular phosphorylation without affecting the entry of pyridoxal across the cell membrane. 2) A pyridoxal-kinase deficient mutant (HN1) derived from the strain KG980 showed a low ability to take up pyridoxal because of the failure to accumulate it effectively as phosphorylated derivatives. The carrier-mediated nature of pyridoxal uptake, previously suggested by saturation kinetics, was further supported by the present finding that 4'-deoxypyridoxine inhibited pyridoxal uptake competitively, decreasing the intracellular appearance of unmetabolized pyridoxal. It is therefore most likely that pyridoxal enters the cells by facilitated diffusion and is accumulated by conversion to phosphorylated derivatives. Similar results on the uptake of pyridoxine and pyridoxamine are also presented. PMID- 6270290 TI - Odontodysplasia, report of a case. PMID- 6270289 TI - Nitrogen balance and hepatic gluconeogenesis in rats fed on diets containing various proportions of carbohydrate and fat. AB - This study was conducted to investigate the effects of various proportions of dietary carbohydrate and fat in diet on protein and carbohydrate metabolism. The growth rate, nitrogen balance, urinary and serum urea levels, and the activities of key ureogenic and gluconeogenic enzymes in the livers were examined in the weanling and growing rats. The rats were raised on a 20% casein diet containing various proportions of carbohydrate and fat, viz. 30% and 50%, 50% and 30%, 60% and 20% or 70% and 10% as calorie percent, respectively, for 10 days. For both weanling and growing rats, the growth rate was unaffected by the alteration in the proportions of carbohydrate and fat in the diets. However, in the rats fed on the 30% carbohydrate-50% fat diet, the urinary excretion of nitrogen and urea were reduced in both groups and these findings were reflected in the reduced serum urea level. Arginase activity decreased. In contrast, glucose-6-phosphatase activity was enhanced in the animals of the 30% carbohydrate-50% fat diet group as compared to the other groups. These results suggest that a low carbohydrate high fat diet causes the reduction of urea formation and the enhancement of glucose formation at a fixed level of protein in the diets in weanling as well as in growing rats. PMID- 6270291 TI - [Studies on expression of the differentiated phenotype of chondrocytes in culture. 1. Effects of parathyroid hormone and cyclic AMP on the activity of ornithine decarboxylase and expression of the differentiated phenotype of chondrocytes in culture]. PMID- 6270292 TI - [Possible existence of enkephalin neurons in the striopallidal pathway in rat brain]. PMID- 6270293 TI - The ultrastructural features of malakoplakia. AB - The ultrastructural features of a case of disseminated malakoplakia are described and compared with available published cases. Generally macrophages were involved (although characteristic inclusions were seen in association with some plasma cells) and contained three types of cellular inclusions. These were (1) the phagolysosomes (2) and intermediate structure sharing features of the phagolysosome and the Michaelis-Gutmann (MG) body and (3) the MG body. All inclusions were delineated by unit membranes and their matrices (except those of large MG bodies) composed of membranous whorls and loops. It is suggested that these inclusions represent stages in the development of the MG body. Septate junctional complexes were observed between phagolysosomes and small MG bodies. These complexes may represent either a mechanism of organelle fusion or an abnormality of molecular organisation of the limiting unit membrane. PMID- 6270294 TI - The enzyme histochemistry of lymphoid and non-lymphoid cells of the human palatine tonsil: a basis for the study of lymphomas. AB - The distribution of various hydrolytic enzymes has been determined in 27 human palatine tonsils by means of conventional enzyme histochemical techniques, and lysozyme (muramidase) activity has been localised in eight tonsils by the unlabelled antibody peroxidase-antiperoxidase complex (PAP) method. The enzyme activities of various cell types are compared and the effect of various methods of fixation and processing discussed. The results suggest that arrangement of various histiocytic cells types within the tonsillar follicles and crypt epithelium is related to the processing of antigen. The PAP method for lysozyme demonstrates a smaller population of cells than is demonstrated by the alpha naphthyl acetate esterase (ANAE) method. T-cells are demonstrated by the presence of dot-like ANAE activity in their cytoplasm. Large numbers of the lymphocytes of this type were located in the T-dependent areas of the tonsil, and are frequent beneath the crypt epithelium. The efferent lymphatic vessels appeared to contain an almost pure population of T-cells. The immunohistochemical method for lysozyme did not differentiate between T- and B-cell areas, dot-like activity being absent. As in other workers' studies on non-lymphoid cells of the murine spleen, several types of glass-adherent cells have been identified in short-term cell cultures from the human tonsil. True dendritic cells and branching macrophages differ in several ways (as in the mouse spleen). The tonsil is considered to be a useful control "reactive" lymphoid organ, to act as a baseline tissue in an extended study of morphology and enzyme histochemistry in the lymphomas. PMID- 6270295 TI - Evidence that cannabidiol does not significantly alter the pharmacokinetics of tetrahydrocannabinol in man. AB - The pharmacokinetics of delta 9-tetrahydrocannabinol (THC) administered intravenously was evaluated in four subjects after oral administration of placebo and 1500 mg of cannabidiol (CBD) according to a crossover design. The cannabidiol pretreatment had no apparent effect on THC pharmacokinetics, yet there may have been minimal effect on the formation and excretion of metabolites. The total (metabolic) blood clearance of THC averaged 17.4 ml/min/kg without CBD and 20.9 ml/min/kg with CBD, and was probably hepatic blood flow limited. The apparent steady-state volume of distribution averaged 9.86 (with CBD, 10.54) liters/kg. Irrespective of CBD pretreatment, the renal clearance of THC metabolites ranged from 17 ml/min after approximately 1 hr to 1.13 ml/min 3.5 days after dosing with THC. The apparent terminal halflife for metabolites averaged 8.2 days. PMID- 6270296 TI - The effect of chronic lithium treatment on rat pineal N-acetyltransferase rhythm. AB - Lithium chloride administration to rats for 5 weeks caused a significant decrease in dark-induced activity of rat pineal N-acetyltransferase (E.C.2.3.1.5.). This effect is not observed after 3 days or 3 weeks of treatment. Furthermore, chronic lithium treatment suppressed the amplitude and may have delayed the peak of the diurnal cycle of N-acetyltransferase activity. In vitro, various concentrations of lithium chloride (2, 4 and 10 microM) did not affect N-acetyltransferase activity. Beta adrenergic receptor binding studies with [3H]dihydroalprenolol indicated a decrease in number of pineal beta adrenergic receptors in rats treated chronically with lithium. These results are consistent with a lithium induced subsensitivity of pineal beta adrenergic receptors and may explain the effect of lithium on pineal-mediated cyclic behaviors. PMID- 6270297 TI - Relationship between cyclic guanosine 3':5'-monophosphate formation and relaxation of coronary arterial smooth muscle by glyceryl trinitrate, nitroprusside, nitrite and nitric oxide: effects of methylene blue and methemoglobin. AB - The purpose of the present study was to determine time course relationship between cyclic GMP accumulation and relaxation in bovine coronary artery and evaluate the effects of recently identified inhibitors, methylene blue and methemoglobin, on these relationships. Arterial strips were suspended in specially mounted tissue baths which permitted continuous recording of isometric tension until rapid freeze-clamping for subsequent determination of cyclic GMP levels by radioimmunoassay. Relaxation and cyclic GMP levels were measured in submaximally contracted strips at zero time (untreated) or 5-sec to 5-min intervals after exposure to 0.5 microliter of nitric oxide, 1 microM glyceryl trinitrate, 1 microM sodium nitroprusside of 1 mM sodium nitrite in the absence or presence of 10 mM methylene blue or 1 microM methemoglobin. Cyclic GMP accumulation preceded onset of relaxation elicited by nitric oxide and glyceryl trinitrate and temporally correlated with relaxation induced by sodium nitroprusside and sodium nitrite. Methylene blue simultaneously inhibited cyclic GMP accumulation and relaxation induced by all four relaxants. In contrast to methylene blue, methemoglobin abolished cyclic GMP accumulation and relaxation elicited by nitric oxide without altering responses to glyceryl trinitrate, sodium nitroprusside and sodium nitrite. These findings are consistent with and strongly support an involvement of cyclic GMP formation in vascular smooth muscle relaxation elicited by nitrogen oxide-containing vasodilators. PMID- 6270298 TI - Studies on the interaction of propranolol and tetrodotoxin on dV/dtmax of canine Purkinje fiber action potentials. PMID- 6270299 TI - The binding and analgesic properties of a sigma opiate, SKF 10,047. PMID- 6270300 TI - Evidence against the presence of presynaptic inhibitory adenosine receptors in the cat nictitating membrane. PMID- 6270301 TI - Electrophysiological response of cerebellar Purkinje neurons to leukotriene D4 and B4. AB - The biological effects of leukotrienes B4 and D4, two recently characterized products of arachidonic acid metabolism, were studied on the guinea-pig ileum in vitro and on rat cerebellar Purkinje cells in vivo. Leukotriene D4 induced contraction of the ileum and a prolonged excitation of Purkinje cells. Both of these effects were antagonized by the "slow reacting substance" end-organ antagonist, FPL 55712. Leukotriene B4 had no central electrophysiological effects at the doses used and no effects on the guinea-pig ileum at doses below 0.2 microM. Taken together with previous studies, these data suggest that leukotrienes with slow reacting substance activity may modulate the excitability of central neurons as well as peripheral smooth muscle tone. PMID- 6270302 TI - Alpha adrenergic receptor subtypes in rabbit uterus: mediation of myometrial contraction and regulation by estrogens. PMID- 6270303 TI - Endogenous prostacyclin contributes to the efficacy of a thromboxane synthetase inhibitor for preventing coronary artery thrombosis. PMID- 6270305 TI - Adenosine regulation of cyclic 3',5'-adenosine monophosphate formation in rat spinal cord. AB - Adenosine stimulates the formation of cyclic 3',5'-adenosine monophosphate (cyclic AMP) in rat spinal cord tissue slices in a concentration-dependent manner with maximal accumulation (30 pmol/mg of protein) at a concentration of 1 mM (Ec50 50 microM). 2-Chloroadenosine (EC50 1 microM) produced a maximal accumulation to 50 pmol/mg of protein. The adenosine antagonists, theophylline and isobutylmethylxanthine, block the increase, and the phosphodiesterase inhibitor, RO 20-1724, potentiates the increase in cyclic AMP accumulation. Adenosine deaminase eliminated the adenosine-dependent increase. Cyclic AMP accumulation was also enhanced by ATP, ADP and 5'-AMP. However, the stimulation due to these nucleotides was dependent upon conversion to adenosine. The increase in cyclic AMP accumulation was more than additive when adenosine was combined with norepinephrine. This potentiation effect is blocked by theophylline, isobutylmethylxanthine and alpha adrenergic antagonists. Additional experiments revealed that only postsynaptic alpha adrenergic agonists were capable of potentiating the response to adenosine. The interaction is concentration dependent, is also observed with phosphorylated nucleotides of adenosine and is blocked specifically by alpha receptor antagonists. Receptor binding assays revealed that adenosine does not alter the number of alpha adrenergic receptors. Both the alpha receptor and adenosine-stimulated cyclic AMP accumulation were Ca++-dependent. These results suggest that adenosine-dependent cyclic AMP formation in rat spinal cord is mediated through two types of receptors, one of which is independent, and the other coupled to the alpha adrenergic receptor. PMID- 6270304 TI - Alpha adrenergic-mediated renin release is prostaglandin-dependent. AB - The contribution of the alpha adrenergic receptor activation to the sympathetic mediated renin release was examined by utilizing intrarenal phenylephrine infusion in beta adrenergically blocked anesthetized mongrel dogs. Increasing doses of intrarenal phenylephrine resulted in a dose-dependent decrease in renal blood flow, a dose-dependent decrease in urinary excretion of sodium and a stimulation of renal renin release which was not dose-dependent. The effect of intrarenal phenylephrine on renin release was totally abolished by pretreating the dogs with indomethacin, even though neither the vasoconstriction nor the decrement in urinary sodium excretion was altered by the drug. In addition, intrarenal phenylephrine did not elicit renal renin release in dogs with kidneys that were rendered nonfiltering. These data are consistent with the hypothesis that exogenous renal alpha adrenergic stimulation in the dog contributes to renin release through the prostaglandin system and that the stimulation of the macula densa mechanism is responsible for the major effect of phenylephrine on renin release. PMID- 6270306 TI - Renal alpha-1 and alpha-2 adrenergic receptors: biochemical and pharmacological correlations. AB - [3H]Dihydroergocryptine, a nonselective alpha adrenergic antagonist, the alpha-1 selective antagonist, [3H]prazosin and the alpha-2 selective antagonist, [3H]yohimbine, were used to study binding sites in rat renal membranes. To establish a correlation between binding and a biological function, the ability of alpha adrenergic agents to stimulate or inhibit vasoconstriction was quantified in vitro using an isolated perfused kidney preparation. Binding with each radioligand was rapid, saturable and specific. Moreover, the order of potencies of a variety of adrenergic agents, determined by competitive inhibition studies, suggested that the binding of each radioligand was to sites with alpha adrenergic specificity. The total number of binding sites in these rat renal membranes. determined with [3H]dihydroergocryptine (Bmax, 212 fmol/mg of protein; KD, 12.8 nM) was approximately equal to the sum of binding site concentrations determined with the alpha-1 and alpha-2 selective radioligands (Bmax, 57 and 170 fmol/mg of protein; KD, 0.85 and 20 nM, respectively). However, the alpha receptor mediating renal arteriolar vasoconstriction appeared to be of the alpha-1 subtype as there was a close correlation between the in vitro results and the binding data determined with [3H]prazosin (r = 0.93). In addition, in both the functional and [3H]prazosin binding studies, unlabeled prazosin ws 5 to 40-fold more potent than the nonselective antagonist, phentolamine, and 400- to 1500-fold more potent than the alpha-2 antagonist, yohimbine. These studies suggest that rat renal plasma membranes contain binding sites with both alpha-1 and alpha-2 adrenergic receptor specificity, in a ratio of approximately 1:3. Despite the preponderance of alpha 2 receptors, the alpha receptor mediating renal vasoconstriction appears to be of the alpha-1 type. PMID- 6270307 TI - Heavy metal inhibition of p-aminohippurate transport in flounder renal tissue: sites of HgCl2 action. AB - p-Aminohippurate (PAH) uptake by teased flounder renal tubules was concentrative, Na-dependent, dependent on aerobic metabolism and inhibited by competitor organic anions and ouabain. Dose-response data for ouabain inhibition of PAH transport and tubule Na,K-adenosine triphosphatase (ATPase) activity were identical. In ouabain-treated tubules, reductions in PAH uptake correlated with alterations in total tissue Na and K even though both were not affected during the first 5 to 10 min of ouabain exposure. No such delay was found with HgCl2 and, as with ouabain, reductions in transport correlated well with alterations in tissue Na and K. Tissue respiration data indicated that low concentrations of HgCl2 rapidly affected the Na,K-ATPase. With higher HgCl2 concentrations, intracellular metabolic sites appeared to be affected. Transport studies with ouabain-poisoned tubules and plasma membrane vesicles indicated that the energetically uncoupled PAH carrier(s) were affected by HgCl2, but carrier sensitivities to Hg were lower than for the Na,K-ATPase. The data indicate that HgCl2 inhibits PAH transport primarily by reducing ion gradients that drive PAH transport. Both inhibition of Na,K-ATPase and increases in membrane permeability to Na and K appear to be involved. PMID- 6270308 TI - Alpha and beta adrenergic effects of the stereoisomers of dobutamine. AB - Dobutamine and its stereoisomers were evaluated for alpha and beta adrenergic activities in vitro. The racemate and the (--)-isomer were found to be potent partial agonists of alpha adrenergic receptors in rat aorta. The (+)-isomer lacked alpha agonist activity. The affinities of (+/-)-, (+)- and (--)-dobutamine for the alpha adrenergic receptor were high (--log KB values of 7.01, 7.02 and 7.07, respectively) and not significantly different from one another. These data indicate that (+)- and (--)- dobutamine bind equally to the alpha adrenergic receptor, however, subsequent to binding, only the (--)-isomer is capable of activating the receptor and eliciting an alpha adrenergic response. The (+) isomer, which has the same affinity as the (--)-isomer but which lacks agonist activity, is a potent competitive alpha blocker. Both stereoisomers of dobutamine were agonists of beta adrenergic receptors of cat papillary muscle and right atria. In contrast to the alpha adrenergic effects, the more potent isomer at the beta adrenergic receptor was (+)- dobutamine. The isomeric activity ratio for the stereoisomers of dobutamine was approximately 1 log unit in favor of the (+) isomer with respect to both inotropic and chronotropic responses. Dose-response curves to the racemate were always situated between the stereoisomers, approximately 2-fold to the right of (+)-dobutamine. These results indicate that the stereochemical requirements of alpha and beta adrenergic receptors are opposite for the stereoisomers of dobutamine with the alpha receptor favoring the (--)-isomer and the beta receptor favoring the (+)-isomer. PMID- 6270309 TI - Fluoride inhibition of the hydro-osmotic response of the toad urinary bladder to antidiuretic hormone. AB - The hydro-osmotic response of the toad bladder to antidiuretic hormone and cyclic AMP was inhibited by the methoxyflurane metabolite, fluoride. The osmotic transfer of water in the absence of hormone was unaffected by fluoride as was the hydroosmotic response due to hypertonicity of the serosal bathing media. Osmotic water movements across N-ethylmaleimide-"fixed" vasopressin or cyclic AMP stimulated bladders were likewise unchanged by fluoride, suggesting that fluoride is exerting an action subsequent to the endogenous formation of cyclic AMP but before the final effector mechanism. Fluoride increased intracellular cyclic AMP concentrations even in the presence of added hormone. Fluoride suppressed calmodulin activity and prevented its activation of phosphodiesterase. Fluoride had no effect on oxygen consumption of toad urinary bladder cells but reduced lactate formation and anerobic metabolism. This decrease in the glycolytic energy source did not contribute to the inhibition of the hormonal response since 2 deoxyglucose was without effect on hormonal mediated osmotic-water flow. It is postulated that the fluoride-induced polyuria after methoxyflurane anesthesia may be due in part to the ability of fluoride to interfere with calcium and calmodulin-initiated processes (other than phosphodiesterase activity) that may occur in the stimulus-reabsorption coupling response of antidiuretic hormone. PMID- 6270310 TI - Kinetics of sodium channel modification by the insecticide tetramethrin in squid axon membranes. PMID- 6270311 TI - Cyclic AMP, morphine, met-enkephalin and neuronal firing. AB - Extracellular recordings were made of spontaneous neuronal firing and of nociceptive stimulus-evoked neuronal firing in the mesencephalic reticular formation of the rat. Microiontophoretically administered morphine and met enkephalin blocked the nociceptive stimulus-evoked neuronal firing of some neurons in the mesencephalic reticular formation; naloxone antagonized the effects of morphine and met-enkephalin. In neurons in which morphine and met enkephalin blocked the nociceptive stimulus-evoked firing, the microiontophoretic administration of dibutyryl cyclic AMP (cAMP), 8-bromo cAMP and Ro 20,1724 (a phosphodiesterase inhibitor) consistently (96%) reversed this blockade of evoked firing. The effects of dibutyryl cAMP were specific, because butyrate and 5'-AMP, possible metabolites of the cAMP analog, reversed the blockade by morphine and met-enkephalin of the nociceptive stimulus-evoked neuronal firing much less frequently (29%). These result support the hypothesis that the occupation of opiate receptors triggers an inhibition of the enzyme, adenylate cyclase, as a mechanism of action. The cAMP analogs excited the spontaneous firing in only 60% of the neurons in which they reversed the opioid blockade of pain-evoked firing. This suggests that the mechanism of action of the cAMP analogs on spontaneous firing may differ from that on pain-evoked firing. PMID- 6270312 TI - Pre- and postsynaptic alpha adrenoceptor selectivity studies with yohimbine and its two diastereoisomers rauwolscine and corynanthine in the anesthetized dog. AB - The selectivity of yohimbine and its two diastereoisomers rauwolscine and corynanthine for pre- and postsynaptic alpha adrenoceptors has been investigated in the anesthetized dog. Antagonism of the inhibitory effect of clonidine on the tachycardia produced by electrical stimulation of the ansa subclavia was used as a measure of presynaptic alpha-2 adrenoceptor blockade. Inhibition of the diastolic pressor response to phenylephrine in ganglion and beta blocked dogs was used as a measure of postsynaptic alpha-1 adrenoceptor blockade. All three of the isomers reduced, and at higher doses reversed, the inhibitory effect of clonidine Yohimbine and rauwolscine were equipotent in this respect and were approximately 100-fold more potent than corynanthine. However, all the isomers were equipotent as antagonists of the diastolic pressure response to phenylephrine. Yohimbine and rauwolscine were approximately 30 times more potent as alpha-2 adrenoceptor than alpha-1 adrenoceptor antagonists, whereas corynanthine was 10-fold more potent at alpha-1 adrenoceptors than at alpha-2 adrenoceptors. These results are in broad agreement with those previously reported from in vitro experiments showing yohimbine and rauwolscine to be preferential alpha-2 adrenoceptor antagonists and corynanthine to be a preferential alpha-1 adrenoceptor antagonist. It is concluded that the high affinity of the antagonists yohimbine and rauwolscine for alpha-2 adrenoceptors is responsible for their selectivity because at the level of blockade of postsynaptic alpha-1 adrenoceptors both isomers were equipotent with corynanthine. PMID- 6270313 TI - Functional opiate receptors in the guinea-pig ileum: their differentiation by means of selective tolerance development. AB - A differentiation of multiple opiate receptors in the electrically stimulated longitudinal muscle-myenteric plexus preparation of the guinea-pig ileum is described employing the technique of tolerance development of selective opiate receptors. Apparently, mu- and kappa-opiate receptors are clearly differentiated in this tissue, since highly tolerant mu-receptors (80-fold) coexist with kappa receptors of almost normal sensitivity and vice versa. Attempts to demonstrate delta-receptors failed, as there was complete cross-tolerance between mu- and delta-agonists. It is proposed that delta-agonists bring about their action via mu-receptors in this preparation. Although mu- and kappa-receptors have been demonstrated, neither of them appear to represent a uniform population. PMID- 6270314 TI - Effects of troleandomycin administration on cholesterol 7 alpha-hydroxylase activity and bile secretion in rats. AB - Repeated administration of troleandomycin increased bile flow but decreased the biliary secretion of bile acids in rats. The increased bile flow was associated with a parallel increase in the biliary clearance of [14C]erythritol. Analysis of the relationship between bile flow and bile acid secretion indicated that, for any given rate of bile acid secretin, bile flow was higher in troleandomycin treated rats than in control rats. The increased bile flow was associated with an increased activity of Na+,K+-adenosine triphosphatase in liver plasma membranes. The decreased bile acid secretion into bile was associated with a similar decrease in the bile acid pool size, a decreased bile acid synthesis rate and a decreased activity of microsomal cholesterol 7 alpha-hydroxylase. The concentration of bile acids in serum, the hepatic extraction ratio of [3H]taurocholate and its biliary transport maximum were not modified. It is concluded that repeated administration of troleandomycin increases the canalicular bile acid-independent flow but decreases the activity of cholesterol 7 alpha-hydroxylase, the synthesis, the pool size and the biliary secretion rate of bile acid in rats. PMID- 6270315 TI - Effects of rubidium on contractility and sodium pump activity in guinea-pig ventricle. AB - Inhibition of the Na+-K+ active transport system has been postulated to be one mechanism through which myocardial contractility is increased. Rubidium is one substance which has been shown to increase the contractility of guinea-pig atria and inhibit the activity of the isolated Na+,K+-adenosine triphosphatase of guinea-pig ventricle. A reexamination of these results confirmed the positive inotropic effect of rubidium on guinea-pig atria and demonstrated that this effect on contractility is accompanied by a decrease in both resting potential and action potential duration. However, it was also found that rubidium produced a transient negative inotropic effect in guinea-pig ventricle. The latter response was closely paralleled by a transient shortening of action potential duration. A concentration of rubidium maximally effective in decreasing contractility (2.0 mM) had no effect on the slow response action potential or contraction. RbCl (0.1 mM) had no effect on cyclic adenosine 3':5'-monophosphate levels of the ventricle or atrium. RbCl did inhibit active transport in the ventricle, as evidenced by a significant reduction in the electrogenic contribution on the active transport system to the maximal diastolic membrane potential during high-frequency drive. These results demonstrate that RbCl has different effects on the contractility of atrial and ventricular muscle. They also suggest that inhibition of the sodium pump is not necessarily accompanied by an increased force of myocardial contraction. PMID- 6270316 TI - Interaction between impulse-flow and delta 9-tetrahydrocannabinol within the septal-hippocampal cholinergic pathway of rat brain. AB - The role of the cholinergic cell bodies of the medial septum in the transsynaptic reduction of nerve activity elicited by delta 9-tetrahydrocannabinol (THC) in the septal-hippocampal cholinergic pathway was assessed by measuring the ability of delta 9-THC to reduce sodium-dependent high affinity choline uptake in rat hippocampus 2 hr after electrolytic destruction of the cholinergic cell bodies in the septum. delta 9-THC did not reduce choline uptake in rats with electrolytic lesions of the cholinergic cell bodies in the septum. Because lesioning of cholinergic cell bodies itself reduces impulse-flow in cholinergic neurons, this observation has two interpretations: either delta 9-THC acts transynaptically at the cholinergic cell bodies in the septum, or alternatively, requires impulse flow in the septal-hippocampal cholinergic pathway in order to reduce its activity. To test if delta 9-THC required impulse-flow in order to reduce the activity or the septal-hippocampal cholinergic pathway, the ability of delta 9 THC to reduce choline uptake in hippocampus of rats with cingulate bundle transections, another surgical procedure which reduces impulse-flow iun the septal-hippocampal cholinergic pathway, was measured. In cingulate bundle transected rats, delta 9-THC did not reduce choline uptake in hippocampus. Taken together with the effects of delta 9-THC in septal-lesioned rats, this observation favors the interpretation that delta 9-THC requires impulse-flow in the septal-hippocampal cholinergic pathway in order to reduce the activity of this pathway. PMID- 6270317 TI - Enhancement of the 5-HT neurotransmission by antidepressant treatments. AB - The hypothesis of an etiopathogenic role of 5-HT and that of a mediation by the 5 HT system in the effect of antidepressant treatments have often been confused. Little unequivocal evidence exists for a 5-HT deficit in depression. However, several recent animal and clinical data suggest that the 5-HT system might contribute to the therapeutic effect of various antidepressant treatments. Long term administration of tricyclic antidepressant (TCA) drugs induces a sensitization of rat forebrain neurons to iontophoretically-applied 5-HT. Repeated electroconvulsive shocks result also in an increased sensitivity of forebrain 5-HT receptors. However, chronic administration of a new antidepressant drug, zimelidine, a potent and long-lasting 5-HT uptake blocker, fails to modify 5-HT receptor sensitivity. These results suggest that enhancement of 5-HT neurotransmission obtained via either pre- or postsynaptic mechanisms might determine the antidepressant effect of these treatments. In a recent clinical study, we observed that lithium administration to TCA-resistant depressive patients induced a rapid relief of depression. It is possible that the presynaptic enhancing effect of lithium on the 5-HT system might unveil the TCA induced sensitization of the postsynaptic 5-HT receptors. Most depressed patients exhibit marked diurnal variations of mood. Preliminary experiments in rats revealed that the responsiveness of hippocampal neurons to iontophoretically applied 5-HT is enhanced in the evening. Similar diurnal variations of 5-HT receptor sensitivity might occur in human brain and be related to diurnal variation of mood in depression. Since normal individuals do not show these fluctuations of mood, it is proposed that the "mood regulating system" might become 5-HT dependent in depressed patients. PMID- 6270319 TI - [Kinetic study of serotonin metabolism in the suprachiasmatic nucleus of the rat: neuroendocrine incidence (author's transl)]. AB - The suprachiasmatic nucleus (SCN), which is implicated in the regulation of several cyclic neuroendocrine rhythms, displays a conspicuous and apparently specific serotoninergic innervation. Our study was intended to establish more precise correlations between the incidence of serotonin (5 HT) metabolism in the activity of the SCN, and neuroendocrine rhythms. For this purpose, castrated female rats, having subcutaneous implants of estradiol, were studied. These animals display very marked circadian fluctuations in plasma levels of the pituitary hormones ACTH, LH and PRL; a relatively well-synchronized increment in all these hormones occurs between 11.00 and 15.00. Punches were obtained to determine the endogenous content of 5 HT, measured by a radioenzymatic technique, simultaneously in SCN and median eminence (ME). An index of SCN activity was determined from in vivo SCN 2-deoxy (1-14C) glucose (DG) uptake; the retention was estimated on radioautographs. Endogenous level of 5 HT displayed a marked circadian rhythm with a peak between 12.00 and 15.00 in the SCN; 5 HT levels were constant throughout the day in the ME. 14C-DG uptake was greater at 15.00 than at 9.00. However, after PCPA treatment or raphe lesions, the uptake of 14C-DG was the same at 9.00 and 15.00. Taken together, these results strongly suggest that in our model: (1) SCN displays a rhythm of activity in the light period; (2) SCN displays specific rhythms in the content of 5 HT; (3) the SCN rhythm of activity must be under an inhibitory control of 5 HT, since the lowest metabolic level is increased at 9.00 by inactivation of 5 HT system; (4) the close relationships between the initial release phase of several pituitary hormones, the increase of metabolic activity in the SCN and the higher level of the 5 HT in the nucleus suggest that 5 HT terminals in the SCN play an important part in the control of cyclic hormonal secretion. PMID- 6270318 TI - [Are bulbo-spinal serotonergic systems involved in the detection of nociceptive messages? (author's transl)]. AB - Intensely noxious peripheral stimuli of the anaesthetized rat produce two changes in the activity of convergent dorsal horn units: the segmental neuronal pool is activated, while all other convergent neurones are inhibited. These Diffuse Noxious Inhibitory Controls (DNIC) are highly potent (60-80% inhibition) and suppress all convergent neuronal activity, whether spontaneous or evoked by noxious or nonnoxious stimuli. On the other hand, they have no effect on other dorsal horn cell types, including noxious-only and proprioceptive units. The "DNIC" circuits include at least one supraspinal relay since DNIC is not seen in spinal animals. Furthermore, they are greatly reduced by lesions of the Nucleus Raphe Magnus (NRM). It has been shown that this nucleus massively projects onto the spinal cord, in particular onto the dorsal horn, and that stimulation of the NRM results in convergent unit inhibition of the same degree of magnitude as with DNIC. The role of serotonergic mechanisms in DNIC can be demonstrated pharmacologically: pCPA pre-treatment (3 daily I.P. injections, 300 mg/kg) or cinanserin (4 mg/kg I.V.) both result in a potent decrease (50-80%). We have proposed that the nociceptive message from the convergent units could result in a contrast between activity of the activated segmental pool and silence of the remaining convergent units. If this hypothesis can be verified, then some raphe nuclei and brain stem serotonergic pathways may function as filters in the detection of nociceptive messages, allowing extraction of information from somatic background activity including the firing from peripheral mechanoreceptors. While superficially paradoxical in fact our hypothesis fits well with the observation of profound analgesia following NRM stimulation: indeed, this hypothetical contrast would be completely eliminated by NRM stimulation since both neuronal pools would then be inhibited. PMID- 6270320 TI - Cyclic nucleotides and calcium in human lymphocytes induced to divide. AB - Intracellular concentrations of cyclic adenosine 3'-5' monophosphate (cAMP) and cyclic guanosine 3'-5' monophosphate (cGMP) were measured in human lymphocytes induced to divide by the addition of lectins, 12-O-tetra-decanoylphorbol-13 acetate (TPA) and the calcium ionophore A 23187. cGMP levels rose within minutes without concomitant alterations in cAMP concentration. The cAMP and cGMP levels rose during the prereplicative and replicative phases respectively. Under calcium depleting conditions, both the fluctuations in cyclic nucleotide levels and the increase in [3H[ thymidine incorporation into DNA were abolished, suggesting a role for calcium ions in the regulation of lymphocyte proliferation. PMID- 6270321 TI - [Autoradiographic and cytochemical localization of (Na+-K+)-activated adenosine triphosphatase in the acini of dog salivary gland (author's transl)]. AB - The distribution of Na pump sites (Na+-K+ ATPase) in the acinar cells of dog submandibular gland was demonstrated by light and electron microscopical radioautography of 3H-ouabain binding sites and electron microscopical ATPase cytochemistry. The grains of 3H-ouabain by light microscopical radioautography were localized to the basal parts of acini and/or the striated ducts, and a small quantity of the grains was also present on the luminal parts of acini. The grains of 3H-ouabain by electron microscopical radioautography and the reaction products of ATPase were found to be localized on the basolateral plasma membrane of both serous and mucous cells, while slightly on the microvilli of the luminal plasma membranes. The present evidence that the distribution of ATPase on the acinar cells determined by the cytochemistry is well concomitant with that of 3H-ouabain binding sites on the acinar cells by the radioautography, suggests that the above mentioned ATPase is Na+-K+ ATPase, a Na pump. The relationship of the distribution of the Na+-K+ ATPase and the cation transport of the plasma membranes in the acinar cells of the dog submandibular gland are discussed. PMID- 6270322 TI - Malignant haemangiopericytoma presenting as an insulinoma. PMID- 6270323 TI - Survival of clonogenic murine epithelioma cells exposed to 52 MeV proton beam produced from the cyclotron at INS. PMID- 6270324 TI - Inactivation of cultured human tumor cells irradiated by cyclotron neutrons and protons. PMID- 6270325 TI - Cross placental transfer of 198Au-colloid in near term rats. PMID- 6270326 TI - Effects of daily treatment with a radioprotector WR-2721 on Ehrlich's ascites tumors in mice : suppression of tumor cell growth and earlier death of tumor bearing mice. PMID- 6270327 TI - The regulation of progesterone receptor by 17 beta estradiol and tamoxifen in the Zr-75-1 human breast cancer cell line in defined medium. AB - The regulation of progesterone receptor by 17 beta estradiol and tamoxifen in the ZR-75-1 human breast cancer cell line in defined medium is described. ZR-75-1 cells maintained in serum free hormone supplemented medium minus estradiol lack progesterone receptor activity. Readdition of estradiol to these cells leads to a marked stimulation of progesterone receptor activity (0 to greater than 100 fmols of specifically bound progesterone per million cells). Tamoxifen (10(-6)M-10( 8)M) does not stimulate progesterone receptor activity in this cell line. The presence of progesterone receptor activity is not directly related to growth. Withdrawal of insulin in the continued presence of estradiol has no effect on progesterone receptor concentration although net cell growth ceases. Conversely, withdrawal of estradiol in the continued presence of insulin induces a cessation of net cell growth accompanied by a loss of all progesterone receptor activity within 3-5 days. PMID- 6270328 TI - Plasma membrane enzymes in BALB/c lymphomas with either T or B cell properties, I. 5'-Nucleotidase. AB - Comparison of membrane bound 5'-nucleotidase activity has been made in crude extracts and plasma membrane fractions from Abelson virus transformed lymphomas, IgM, IgG and IgA producing plasmacytomas and from thymomas with different surface antigen markers. 5'Nucleotidase activity was characterised by the following criteria : 1) optimal pH for enzyme activity, 2) specificity of 5'AMP as a substrate at the optimal pH, 3) specific inhibition of the enzyme by alpha beta methylene ADP, 4) inhibition by EDTA. 5'-Nucleotidase was found to be present on the following B cells : Abelson virus transformed lymphomas, plasmacytomas and LPS-stimulated blasts from nu/nu spleens. The enzyme was also found in one thymoma Lut 13, but it was absent from all the other thymomas studied. A possible relationship between 5'-Nucleotidase and purine metabolism in lymphocyte subpopulations is suggested by the results. PMID- 6270329 TI - Inhibition of macrophage phagocytosis after contact with ingestible particles. PMID- 6270330 TI - Whole blood luminol-dependent chemiluminescence. PMID- 6270331 TI - Rabbit synovial tissue in culture: influence of the inflammatory state in vivo on the secretion of collagenase. AB - Primary rabbit adherent synovial cell monolayer cultures were prepared from animals with antigen-induced synovitis. Primary cultures derived from chronically inflamed tissues released elevated levels of collagenase into the culture media compared to cultures derived from either normal or acute inflammatory tissues. No active of latent collagenase activity was detected in media of cultures derived from normal tissues grown under identical conditions. Cultures of acute inflammatory tissues secreted 1/4 to 1/10 the amount of collagenase when compared to cultures from chronically inflamed tissues. Collagenase secreted by cultures from inflammatory tissues was detected only in latent form. Fetal bovine serum enhanced the trypsin activation of latent collagenase produced by these cultures. PMID- 6270332 TI - Coexistence of rheumatoid arthritis in married couples: a search for etiological factors. AB - Epstein-Barr virus (EBV) and the genetic marker HLA-DR4 both appear to play a role in the etiology of rheumatoid arthritis (RA). Antibodies to viral capsid antigen and the early antigen complex of the EB virus as well as the HLA phenotype were studied in 6 families. Both spouses had classical or definite RA but none of their offspring, all adult, had any evidence of RA. At least 1 spouse of each couple had DR4 and this phenotype was found in 4 of the 8 offspring. All parents and their offspring had been infected with EBV. This study supports the association of DR4 and RA. The presence of DR4 in 4 normal adult offspring was not associated with the absence of primary infection or enhanced levels of EBV specific antibodies. PMID- 6270333 TI - Calcium pyrophosphate crystal formation in model hydrogels. II. Hyaline articular cartilage as a gel. AB - We studied calcium pyrophosphate crystal formation in an in vitro cartilage system. Two parallel troughs were excavated in tibial plateau articular cartilage obtained postmortem. One well was filled with solid sodium pyrophosphate, the other with calcium chloride. After incubation for 24 h at either 10 degrees C or 37 degrees C the precipitate band between the troughs was analyzed for the size and nature of crystals present. In subsequent experiments, the cartilage was pretreated by laceration, contusion, trypsin or hyaluronidase denaturation. We found that cartilage denaturation resulted in formation of larger crystals but that the crystal product in all experiments was identical, alpha CaNa2P2O7.4H2O a nonphysiologic crystal. PMID- 6270334 TI - Neuroarthropathy (Charcot joints) in familial amyloid polyneuropathy. AB - Two patients with familial amyloid polyneuropathy (FAP) of Andrade type developed neuroarthropathy (Charcot joints). In 1 patient both knees were affected and in the other patient both ankles. This complication added markedly to the already severe disability and limitation of movement caused by the underlying illness. To the best of our knowledge this is the first description of Charcot joints in FAP. Neuroarthropathy in FAP is probably more common than previously suspected and should be sought in any case of FAP with joint disease. PMID- 6270335 TI - A structure-activity relationship study of spirolactones. Contribution of the cyclopropane ring to antimineralocorticoid activity. AB - A series of spirolactones containing a cyclopropane ring in the molecule was examined for its effects on the mineralocorticoid receptor. The results were compared with those of a similar series of spirolactones in which the cyclopropane ring was replaced by a double bond. Insertion of a double bond or an alpha-cyclopropane ring into the 1,2 or the 6,7 position leads to a reduction in the binding affinity. The pi-bonding system of the beta-cyclopropane ring at C-6 and C-7 does not promote binding to the receptor. The presence of the 6 beta, 7 beta-cyclopropane ring may deter metabolic activation to account for the enhanced in vivo activity. PMID- 6270336 TI - Calcium pyrophosphate deposition disease: a commonly unrecognized entity. PMID- 6270337 TI - Nucleotide sequence of the kanamycin resistance transposon Tn903. PMID- 6270338 TI - Patterns of integration of viral DNA in adenovirus type 2-transformed hamster cells. PMID- 6270339 TI - A reassessment of the structure of Paracoccus cytochrome c-550. PMID- 6270340 TI - Generation and characterization of condensation aerosols of benzo[alpha]pyrene. AB - Condensation aerosols of benzo[a]pyrene (BAP) with particle sizes ranging from 0.1 to 2 micrometers (aerodynamic diameter) were produced and studied. These aerosols were generated in a glove box by direct vaporization of BaP and homogeneous condensation of the vapor. The aerosol concentration ranged from 50 to 700 micrograms/l with aerosol production rates up to 15 mg BaP per minute. The effects of vaporization temperature and flow rate of diluting air on the particle size distribution and aerosol output were studied. The BaP aerosol was produced with relatively constant mass concentration and particle size distribution for more than 5 h. The aerosol was physicochemically and thermally stable. Data on the in vitro dissolution of BaP particles in aqueous solvents and in different dissolution systems suggested that the organic BaP particle does not dissolve in simple aqueous solvents. Proteins, surfactants, or ethyl alcohol enhanced the rate of dissolution of BaP. The rate of dissolution of BaP particles was inversely proportional to particle size. PMID- 6270341 TI - Estimating the hazards of less hazardous cigarettes. II. Study of cigarette yields of nicotine, carbon monoxide, and hydrogen cyanide in relation to levels of cotinine, carboxyhemoglobin, and thiocyanate in smokers. AB - Yields of chemical constituents such as tar, nicotine, CO, and HCN defined by smoking machines are commonly assumed to provide a reasonable indication of the relative hazard associated with smoking a given brand of cigarette. Results reported here suggest that this assumption should be carefully reexamined. A total of 240 subjects, representing a wide range of smoking and brand characteristics, were recruited for an investigation of possible relations between brand yields and exposure (levels of carboxyhemoglobin, breath CO, plasma cotinine, plasma thiocyanate, and saliva thiocyanate). Exposure was highly correlated with consumption (number of cigarettes per day), but their was no correlation between any estimate of exposure and brand yield when level of consumption was held constant. In addition, a comparison of levels of carboxyhemoglobin and plasma thiocyanate for 16 smokers of "low-hazard" and 15 smokers of "high-hazard" cigarette brands revealed little difference between the two groups, even though average cigarette yields differed as much as 2- to 3 fold. A possible explanation for the results may be that current values for average puff volume, duration, and interval differ significantly from those used in programming smoking machines, particularly in the case of brands with low nicotine delivery. PMID- 6270342 TI - Erythrocyte lead-binding protein after occupational exposure. II. Influence on lead inhibition of membrane Na+,K+-adenosinetriphosphatase. AB - Membrane Na+,K+-adenosinetriphosphatase in erythrocytes from three groups of industrially exposed Pb workers (without toxicity, with toxicity associated with high blood Pb levels, and with toxicity associated with low blood Pb levels) was inversely correlated with Pb in the membrane fraction but not significantly correlated with total erythrocyte Pb. This difference was attributable to the proportion of erythrocyte Pb bound to hemoglobin and a Pb-binding protein of molecular weight 10,000. PMID- 6270343 TI - Characterization of polyoma mutants with altered middle and large T-antigens. AB - The viable polyoma mutants dl1013, dl1014, and dl1015 produced shortened middle and large T-antigens. In mouse 3T3 cells, dl1013 and dl1014 grew at normal rates, and dl1015 grew at a reduced rate. dl1015 behaved phenotypically as a double mutant, with deficiencies both in the stimulation of the host cell and the replication of viral DNA. Only the former defect could be complemented by the ts a mutant, which produced a normal middle T-antigen and a temperature-sensitive large T-antigen. This result suggests that middle T-antigen is involved in the induction of cellular DNA synthesis. Of the three mutants, dl1015 alone failed to transform rat fibroblasts to growth in semisolid medium. This defect could not be complemented by the ts-a mutant. Determination of the base sequences of the mutant DNAs showed that dl1013 and dl1014 had overlapping deletions of 21 and 9 base pairs, respectively, and that the dl1015 deletion of 30 base pairs was contiguous to the other mutations on their 3' sides. Analyses of the mutant t antigens showed that all three mutants produced shortened middle T-antigens, whereas only dl1015 large T-antigen was detectably reduced in size. PMID- 6270344 TI - Arrangement of integrated viral DNA sequences in cells transformed by adenovirus types 2 and 5. AB - The organization of the viral DNA sequences in 15 adenovirus-transformed cell lines was analyzed by the Southern blotting procedure. The site of adenovirus integration in the cellular genome was found not to be unique, and the viral DNA sequences involved in integration were not confined to a specific region of the adenovirus genome. Several cell lines showed simple integration patterns that demonstrated the presence of large continuous stretches of viral DNA. In four cell lines, containing sequences from both molecular ends of the viral genome, the left- and right-hand-terminal sequences appeared to be linked to each other. PMID- 6270345 TI - Biochemical analysis of the p30's of N-, B-, and B leads to NB-tropic murine leukemia viruses of BALB/c origin. AB - Previous analysis of the virion proteins of an N- and a B-tropic type C virus of BALB/c mice, of 16 N-tropic recombinants (XLPN viruses) between these viruses, and of eight NB-tropic viruses derived from the B-tropic virus suggested that among these closely related viruses N-, B-, or NB-tropism was associated with the electrophoretic mobility of p30 on sodium dodecyl sulfate-polyacrylamide gels, and thus that p30 might determine this phenotype. To obtain further evidence for the association of structural markers of p30 with N-, B-, or NB-tropism, we have analyzed the p30's of these same viruses by using two-dimensional tryptic peptide mapping and slab gel isoelectric focusing. The results of these analyses suggest that (i) a single peptide unique to the N-tropic virus p30- is present in the p30 of all N-tropic recombinants; (ii) a single peptide unique to the B virus p30 is not present in p30's of the N-tropic recombinants, and this peptide is also absent in p30's of NB-tropic viruses derived from the B-tropic virus; and (iii) p30's of NB-tropic viruses possess a new tryptic peptide not found in the p30 of their B-tropic virus progenitors, and this new peptide is not found in the p30 of the N-tropic virus of BALB/c or the XLPN viruses. These results are consistent with the possibility that p30 may determine the N-, B-, or NB-tropism of murine leukemia viruses. In addition, these studies indicate that some of the N-tropic recombinants have experienced recombination within the p30 gene. PMID- 6270347 TI - Transcriptional and translational mapping of a 6.6-kilobase-pair DNA fragment containing the junction of the terminal repetition and unique sequence at the left end of the vaccinia virus genome. AB - The penultimate EcoRI fragments from the left and right ends of the vaccinia virus genomes were cloned in phage lambda. Heteroduplex analysis and comparison of restriction fragments indicated that the inverted terminal repetition extended 780 base pairs (bp) beyond the EcoRI site or about 9,800 bp from each end of the genome. Detailed physical, transcriptional, and translational maps of the 6,600 bp left penultimate EcoRI fragment were prepared so as to extend previous maps of the 9,000-bp terminal EcoRI fragment. Polypeptides with molecular weights of 6,000 (6K polypeptide), 13,000, 19,000, 21,000, and 60,000 were synthesized in a reticulocyte cell-free system programmed with immediate early RNA (made in the presence of cycloheximide) or early RNA (made in the presence of cytosine arabinoside) and selected by hybridization to immobilized recombinant DNA. A 22K polypeptide was detected as a translation product of late RNA that hybridized to this DNA fragment. A variety of biochemical procedures were used to size and map the mRNA's. Of the five messages that hybridized to this 6,600-bp EcoRI fragment, only the one for the 21K polypeptide was encoded within the inverted terminal repetition and hybridized to the rightward-reading DNA strand. (Three additional early polypeptides were encoded within the first 9,000 bp of the inverted terminal repetition.) The remaining early polypeptides were encoded within the unique portion of the penultimate EcoRI fragment and were transcribed from the leftward-reading strand. Additional high-molecular-weight early RNAs of unknown function were also detected; however, there was no evidence indicating that mature mRNA's were spliced. PMID- 6270346 TI - Cell killing by avian leukosis viruses. AB - Infection of chicken cells with a cytopathic avian leukosis virus resulted in the detachment of killed cells from the culture dish. The detached, dead cells contained more unintegrated viral DNA than the attached cells. These results confirm the hypothesis that cell killing after infection with a cytopathic avian leukosis virus is associated with accumulation of large amounts of unintegrated viral DNA. No accumulation of large amounts of integrated viral DNA was found in cells infected with cytopathic avian leukosis viruses. PMID- 6270348 TI - Extension of the transcriptional and translational map of the left end of the vaccinia virus genome to 21 kilobase pairs. AB - Physical, transcriptional, and translational maps of an EcoRI fragment located between 15,800 and 20,600 base pairs from the left end of the vaccinia virus genome were prepared. Major polypeptides with molecular weights of 14,000 (14K polypeptide), 32,000 and 38,000 were synthesized in a reticulocyte cell-free system programmed with immediate early RNA made in the presence of cycloheximide and selected by hybridization to lambda recombinant DNA containing the EcoRI fragment. With early RNA made in the presence of cytosine arabinoside, an inhibitor of DNA replication, the polypeptide pattern was similar except for quantitative differences in which less 38K polypeptide was detected as a translation product. With late RNA, isolated 6 h after infection without inhibitors, only traces of the early translation products were found and a new 40K polypeptide was detected. The size of the mRNA's for the 14K, 32K, and 38K polypeptides were determined to be approximately 760,880, and 1,150 nucleotides, respectively, by several independent procedures. Several large early RNAs not shown to code for any additional translation products were also detected. The size of the late message for the 40K polypeptide varied from 920 to 3,100 nucleotides. This heterogeneity appeared to be a general property of vaccinia virus late mRNA's. No evidence of RNA splicing was obtained by analysis of RNA DNA hybrids after nuclease S1 treatment. Further analyses using separated recombinant DNA strands and restriction fragments indicated that all mRNA's were encoded by the leftward-reading DNA strand and at least two were overlapping. Since early and late mRNA's were encoded by the same DNA strand, the possibility of temporal regulation by transcriptional strand switching was eliminated. In conjunction with previous studies, a transcriptional map of the left 20,600 base pairs of the vaccinia virus genome was derived. PMID- 6270349 TI - Physical mapping of drug resistance mutations defines an active center of the herpes simplex virus DNA polymerase enzyme. AB - The genome structures of herpes simplex virus type 1 (HSV-1)/HSV-2 intertypic recombinants have been previously determined by restriction endonuclease analysis, and these recombinants and their parental strains have been employed to demonstrate that mutations within the HSV DNA polymerase locus induce an altered HSV DNA polymerase activity, exhibiting resistance to three inhibitors of DNA polymerase. The viral DNA polymerases induced by two recombinants and their parental strains were purified and shown to possess similar molecular weights (142,000 to 144,000) and similar sensitivity to compounds which distinguish viral and cellular DNA polymerases. The HSV DNA polymerases induced by the resistant recombinant and the resistant parental strain were resistant to inhibition by phosphonoacetic acid, acycloguanosine triphosphate, and the 2',3' dideoxynucleoside triphosphates. The resistant recombinant (R6-34) induced as much acycloguanosine triphosphate as did the sensitive recombinant (R6-26), but viral DNA synthesis in infected cells and the viral DNA polymerase activity were not inhibited. The 2',3'-dideoxynucleoside-triphosphates were effective competitive inhibitors for the HSV DNA polymerase, and the Ki values for the four 2',3'-dideoxynucleoside triphosphates were determined for the four viral DNA polymerases. The polymerases of the resistant recombinant and the resistant parent possessed a much higher Ki for the 2',3'-dideoxynucleoside triphosphates and for phosphonoacetic acid than did the sensitive strains. A 1.3-kilobase-pair region of HSV-1 DNA within the HSV DNA polymerase locus contained mutations which conferred resistance to three DNA polymerase inhibitors. This region of DNA sequences encoded for an amino acid sequence of 42,000 molecular weight and defined an active center of the HSV DNA polymerase enzyme. PMID- 6270350 TI - src Genes of ten Rous sarcoma virus strains, including two reportedly transduced from the cell, are completely allelic; putative markers of transduction are not detected. AB - The src genes of different Rous sarcoma virus (RSV) strains have been reported to be highly conserved by some investigators using RNA-cDNA hybridization, whereas others using oligonucleotide, peptide, and serological analyses have judged src genes to be variable in 30 to 50% of the respective markers. Moreover, distinctive src oligonucleotides and peptides of so-called recovered RSVs (rRSV's) whose src genes were reported to be experimentally transduced from the cell are thought to represent specific markers of host-derived src sequences. By contrast, we have pointed out previously that these markers may represent point mutations of parental equivalents. Here we have compared the src-specific sequences of eight RSV strains and of two rRSV's to each other and to a molecular clone of the src-related chicken locus. Our comparisons are based on RNase T(1) resistant oligonucleotides of RNA hybridized to src-specific cDNA, which was prepared by hybridizing RSV cDNA with RNA of isogenic src deletion mutants, or to a cloned cellular src-related DNA. All of the approximately 20 src oligonucleotides of a given RSV strain were recovered by src-specific cDNA's of all other RSV strains or by cellular src-related DNA. The number of oligonucleotides varied slightly with the length of the src deletion used to prepare src-specific cDNA, thus providing a measure for src deletion mutants. Our data indicate that the src genes of all RSV strains tested, including the two reportedly transduced from the cell, are about 98% conserved and completely allelic with only scattered single nucleotide differences in certain variable regions which are subject to point mutations. Hence, based on the src oligonucleotide markers analyzed by us and others, we cannot distinguish between a cellular and viral origin of rRSV's. However, the following are not compatible with a cellular origin of rRSV's. (i) The only putative oligonucleotide marker which is exclusively shared by the two rRSV's studied and which differs from a parental counterpart in a single base was not detectable in cellular src-related DNA. (ii) The number of different allelic src markers observed by us and others in rRSV's was too large to derive from one or two known cellular src-related loci. (iii) The known absence of linkage of the cellular src-related locus with other virion sequences was extended to all non-src oligonucleotides, including some mapping directly adjacent to src. This is difficult to reconcile with the claim that transformation-defective, partial src deletion mutants of RSV which contain both, one, or, as we show here, possibly no src termini nevertheless transduce at the same frequencies, even though homologous, single or double illegitimate recombinations would be involved. Given (i) our evidence that src genes are subject to point mutation under selective conditions similar to those prevailing when rRSV's were generated and (ii) the lack of absolute evidence for the clonal purity of the transformation-defective, partial src deletion mutants of RSV used to generate rRSV's, we submit that the src genes of rRSV's could have been generated by cross-reactivation of nonoverlapping src deletions or mutation of src variants possibly present in transformation-defective, partial src deletion mutants of RSV. To prove experimental transduction, unambiguous markers need to be identified, or it would be necessary to generate rRSV's with molecularly cloned transformation-defective, partial src deletion mutants of RSV. Although our evidence casts doubt on the idea that specific src sequences of rRSV's originated by transduction, the close relationship between viral src and cellular src-related sequences argues that src genes originated at one time in evolution from the cell by events that involved illegitimate recombination and deletion of non-src sequences that interrupt the cellular src locus. PMID- 6270352 TI - Lack of AKR ecotropic provirus amplification in AKR leukemic thymuses. AB - A DNA fragment from the 3' region of a molecularly cloned AKR ecotropic provirus was identified to be specific for the AKR ecotropic murine leukemia virus (MuLV). This selected DNA fragment was used to analyze the integrated MuLV proviruses in normal and leukemic tissue DNAs of AKR mice. In comparison with a DNA fragment from the 5' region of the cloned AKR genome or one representing the entire genome, this selected probe hybridized to only a few MuLV proviruses. By comparing transformed and nontransformed tissue DNAs, it appeared that no amplification of proviral sequences related to the AKR ecotropic MuLV had occurred in thymomas of AKR mice during the development of leukemia in these animals. Analysis of the AKR ecotropic MuLV proviruses revealed a significant degree of polymorphism for these sequences among individuals in the AKR/J strain of mouse. PMID- 6270351 TI - Genomes of murine leukemia viruses isolated from wild mice. AB - The genomes of murine leukemia viruses (MuLV) isolated from wild mice have been studied. Detailed restriction endonuclease maps of the 8.8-kilobase (kb) unintegrated linear viral DNAs were derived for five ecotropic and five amphotropic MuLV's from California field mice, for Friend MuLV, and for one ecotropic and one xenotropic MuLV from Mus musculus castaneus. In general, the California MuLV's were similar in their leftward 6 kb (corresponding to the leftward long terminal repeat [LTR], gag, and pol) and rightward 1 kb (7.8 to 8.8 kb, corresponding to p15E and the rightward LTR). For the region spanning 6.0 to 7.7 kb (which includes the sequences that encode gp70) the amphotropic MuLV's shared few enzyme sites with the ecotropic MuLV's, although the California ecotropic MuLV's were highly related to each other in this region, as were the amphotropic MuLV's. Cross-hybridization studies between amphotropic and California ecotropic MuLV DNAs indicated that they were not homologous in the region 6.3 to 7.6 kb; the California ecotropic viral DNAs cross-hybridized in this region to AKR ecotropic MuLV. When the California viral DNAs were compared with AKR ecotropic viral DNA, many differences in enzyme sites were noted throughout the genome. The U3 regions of the wild mouse LTRs showed partial homology to this region in AKR MuLV. The LTR of Moloney MuLV was highly related to that of the California MuLV's, whereas the LTR of Friend MuLV appeared to be a recombinant between the two types of LTRs. The M. musculus castaneus isolates were most closely related to ecotropic and xenotropic MuLV's isolated from inbred mice. One amphotropic MuLV DNA was cloned from supercoiled viral DNA at its unique EcoRI site in pBR322. Viral DNAs with one and two LTRs were isolated. After digestion with EcoRI, DNAs of both types were infectious. It is concluded that ecotropic and amphotropic MuLV's differ primarily in the region which encodes gp70. PMID- 6270353 TI - Detection and cloning of murine leukemia virus-related sequences from African green monkey liver DNA. AB - By using low-stringency nucleic acid hybridization conditions and specific subgenomic segments of the AKR ecotropic provirus as probes, murine leukemia virus (MuLV)-related sequences were detected in African green monkey (AGM) liver DNA. The MuLV-reactive segments present in restricted AGM DNA ranged from 1.9 kilobases (kb) to greater than 10 kb in size. On the basis of this finding, a 17 kb segment was cloned from a partial EcoRI AGM library in lambda Charon 4A which shared nearly 5 kb of homology with AKR ecotropic MuLV DNA. The MuLV-related sequences detected in restricted preparations of AGM DNA or present in the cloned monkey DNA reacted with probes mapping 2.0 to 7.0 kb from the 5' terminus of the AKR ecotropic provirus. The AGM clone also contained repeated sequences that flanked the MuLV-related segment. Labeled, subgenomic, MuLV-reactive segments of the monkey clone hybridized to multiple restriction fragments of AGM liver DNA, indicating the presence of several copies of the MuLV-related sequences. PMID- 6270354 TI - DNA gyrase inhibitors block development of Bacillus subtilis bacteriophage SP01. AB - SP01 development was inhibited by nalidixic acid and novobiocin in the sensitive host Bacillus subtilis 168M. Inhibition by novobiocin was prevented by a Novr mutation in the cellular DNA gyrase gene. Nalidixic acid inhibition persisted in hosts carrying a Nalr gyrase, but could be overcome by phage mutation. We conclude that SP01 requires for its development subunit B of the host DNA gyrase, but replaces or modifies subunit A. PMID- 6270355 TI - Phosphorylation of the Abelson murine leukemia virus transforming protein. AB - The Abelson murine leukemia virus transforming gene product is a phosphorylated protein encoded by both viral and cellular sequences. This gene product has an amino-terminal region derived from the gag gene of its parent virus and a carboxyl-terminal region of (abl) derived from a normal murine cellular gene. Using a combination of partial proteolytic cleavage techniques and antisera specific for gag and abl sequences, we mapped in vivo phosphorylation sites to different regions of the protein. Phosphoproteins encoded by strain variants and transformation-defective mutants of Abelson murine leukemia virus with defined deletions in the primary sequence of the abl region were compared by two dimensional limit digest peptide mapping. Specific phosphorylation pattern differences for wild-type and mutant proteins probably represented deletions of specific phosphate acceptor sites in the abl region. An in vitro autophosphorylation activity copurified with the Abelson murine leukemia virus protein from transformation-competent strains. A peptide analysis of such in vitro reactions demonstrated that these phosphorylation sites were restricted to the amino-terminal region, and the specific sites appeared to be unrelated to the sites found on proteins phosphorylated in vivo. Thus, the autophosphorylation reaction probably correlates with an activity important in transformation, but the specific end product in vitro bears little resemblance to its function in vivo. PMID- 6270356 TI - Proteolytic enhancement of rotavirus infectivity: molecular mechanisms. AB - The polypeptide compositions of single-shelled and double-shelled simian rotavirus particles were modified by exposure to proteolytic enzymes. Specifically, a major outer capsid polypeptide (VP3) having a molecular weight of 88,000 in double-shelled particles was cleaved by trypsin to yield two polypeptides, VP5* and VP8* (molecular weights, 60,000 and 28,000, respectively). The cleavage of VP3 by enzymes that enhanced infectivity (trypsin, elastase, and pancreatin) yielded different products compared to those detected when VP3 was cleaved by chymotrypsin, which did not enhance infectivity. The appearance of VP5* was correlated with an enhancement of infectivity. Cleavages of the major internal capsid polypeptide VP2 were also observed. The VP2 cleavage products had molecular weights similar to those of known structural and nonstructural rotavirus polypeptides. We confirmed the precursor-product relationships by comparing the peptide maps of the polypeptides generated by digestions with V-8 protease and chymotrypsin. The remaining rotavirus structural polypeptides, including the outer capsid glycoproteins (VP7 and 7a), were not altered by exposure to pancreatic enzymes. Cleavage of VP3 was not required for virus assembly, and specific cleavage of the polypeptides occurred only on assembled particles. We also discuss the role of cleavage activation in other virus specific biological functions (e.g., hemagglutination and virulence). PMID- 6270357 TI - Thymidine kinase not required for antiviral activity of acyclovir against mouse cytomegalovirus. AB - Previous studies of herpesvirus infections have indicated that a virus-specified thymidine kinase is required for the initial phosphorylation of acyclovir [acycloguanosine or 9-(2-hydroxyethoxymethyl)guanine] in the formation of acycloguanosine triphosphate. The latter compound accumulates in infected cells and competitively inhibits the viral DNA polymerase. We found that mouse cytomegalovirus, which does not express a thymidine kinase, was sensitive to the antiviral effects of acyclovir at a 50% inhibitory dose of approximately 0.23 microM. Acyclovir was equally effective against mouse cytomegalovirus in normal 3T3 cells and in 3T3 cells deficient in cellular thymidine kinase. Furthermore, the activity of acyclovir could not be reversed by excess thymidine, which easily reversed the antiviral activity of acyclovir against herpes simplex virus. Using a high-pressure liquid chromatography technique that easily detected acycloguanosine triphosphate in cells infected with herpes simplex virus, we could not detect acycloguanosine triphosphate in mouse cytomegalovirus-infected cells. These experiments demonstrated that the activity of acyclovir against mouse cytomegalovirus is not dependent on a thymidine phosphorylation pathway. Additional experiments are underway to determine whether acycloguanosine triphosphate is produced by another pathway in concentrations sufficient to inhibit mouse cytomegalovirus DNA polymerase. PMID- 6270358 TI - Nonstructural proteins of herpes simplex virus. II. Major virus-specific DNa binding protein. AB - The major herpes simplex virus type 2 DNA-binding infected cell-specific polypeptides 11 and 12 have been purified to homogeneity from extracts of virus infected cells. Monospecific antiserum to the purified protein has been made and used to examine virus temperature-sensitive mutants for defects in the synthesis of the protein and to probe virus DNA synthesis in isolated chromatin. The purified protein acted directly on a polydeoxyadenylic acid-polydeoxythymidylic acid helix, reducing its melting temperature. The results indicated that the protein functions in virus DNA synthesis. PMID- 6270360 TI - Unintegrated viral DNA sequences in a hamster tumor induced by bovine papilloma virus. AB - A fibrosarcoma was induced in a hamster by bovine papilloma virus type 2 (BPV2). The content of BPV2 DNA sequences was measured by DNA-DNA and cRNA-DNA hybridizations. The tumor contained approximately 300 BPV2 genome equivalents per cell. Southern blot hybridization indicated that the viral DNA was in free form, the entire genome most likely being present. In situ hybridization with BPV2 cRNA showed that multiple genome copies were present in each cell. Neither virus particles nor virus coat antigens could be detected in the tumor. A cell line was established from the fibrosarcoma, and the cells contained multiple copies of the BPV2 genome. The latter was in free form, and all of the DNA sequences appeared to be present in multiple copies and in all cells. An extensive search failed to reveal the presence of virus or viral antigens. PMID- 6270361 TI - Demonstration of avian sarcoma virus-coded pp60src in vivo and the anti-pp60src immune response in chickens. AB - The avian sarcoma virus (ASV)-coded transforming protein pp60src was originally detected in vitro in ASV-transformed avian and mammalian cells in experiments involving mammalian antisera to ASV-induced tumors. It is demonstrated here that pp60src is also expressed in vivo in ASV tumors of chickens. Furthermore, the existence of the endogenous pp60src in all chicken cells does not impair the immune response to exogenous pp60src in the chicken. Whereas chicken antibodies can bind to pp60src, they do not serve as substrates for the protein kinase activity of this transforming protein. PMID- 6270359 TI - Construction and isolation of a transmissible retrovirus containing the src gene of Harvey murine sarcoma virus and the thymidine kinase gene of herpes simplex virus type 1. AB - We constructed lambda recombinants containing the Harvey murine sarcoma virus genome and the thymidine kinase (tk) gene of herpes simplex virus type 1 linked to each other. The tk gene was located in a position downstream from both the long terminal repeat and the src gene of Harvey murine sarcoma virus. The DNAs of the lambda recombinants were used to transfect NIH3T3 mouse fibroblasts in order to obtain Harvey murine sarcoma virus DNA-induced foci of transformed cells. The transformed foci were superinfected with a helper-independent retrovirus, and new individual retrovirus were isolated from the superinfected foci. The new viruses could induce focus formation on NIH3T3 cells and could convert NIH3T3(TK-) cells into TK+ cells by carrying the herpes simplex virus type 1 tk gene into the TK- cells. From virus-infected cells, we isolated nonproducer foci on NIH3T3 cells and TK+ transformants on NIH3T3(TK-) cells containing one such new viral genome coding for the dual properties. The new retroviral sequence in the nonproducer cells could be rescued into virus particles at high titers by superinfection with a helper-independent retrovirus. A hybridization analysis indicated that the recombinant virus contained both the Harvey murine sarcoma virus src sequence and the tk gene sequence in a single RNA species approximately 4.9 kilobases long. We concluded that retroviruses can be used as true vectors for genes other than genes that lead to oncogenesis. PMID- 6270362 TI - Growth and persistence of polyoma early region deletion mutants in mice. AB - Replication of early region deletion mutations in polyoma in the kidneys of mice was in most cases reduced by 10- to 10,000-fold, as compared with wild-type polyoma, and the mutants failed to produce the persistent infection observed with wild-type polyoma. PMID- 6270363 TI - Genome analysis of MG virus, a human papovavirus. AB - The genome of MG virus, a variant of the human papovavirus BK virus, consists of two molecules, M1 and M2, M1 and M2 have deletions which correspond to 0.33 to 0.55 map unit and 0.77 to 0.85 map unit, respectively, from the EcoRI site on the BK virus genome. Restriction enzyme analysis of the DNAs of these viruses revealed many alterations in both DNA species. Both M1 and M2 DNAs have three recognition sites for EcoRI. M1 has a single recognition site for HindIII and five sites for PvuII. M2 has a single recognition site for PuvII and three sites for HindIII. The sites of these and several other restriction enzymes on each DNA molecule were mapped after the cloning of M1 and M2 DNAs into pBR322 at their unique HindIII and PvuII sites, respectively. PMID- 6270364 TI - Selective host range restriction of goat cells for recombinant murine leukemia virus and feline leukemia virus type A. AB - We isolated a strain of normal goat fibroblasts which was uniquely selective in that it allowed the replication of xenotropic murine leukemia virus but not polytropic recombinant murine leukemia virus. In addition, feline leukemia virus type A replication was severely diminished in these goat cells, whereas feline leukemia virus type B and feline endogenous RD114-CCC viruses replicated efficiently. No other known cells exhibit this pattern of virus growth restriction. These goat cells allow the study of xenotropic murine leukemia virus in mixtures which also contain recombinant murine leukemia virus and may be helpful in eliminating feline leukemia virus type which often coexists in feline sarcoma or leukemia virus mixtures with other feline leukemia virus types. PMID- 6270365 TI - In vitro transcription catalyzed by heat-treated human rotavirus. AB - The in vitro characteristics of human rotavirus transcription have been examined. The virus has an associated RNA polymerase activity which was activated after a heat shock treatment. The enzyme required the presence of the four ribonucleoside triphosphates and a divalent cation (Mg2+), and it required an optimum pH of 8.5. The polymerase was activated by monovalent salts and inhibited by Na PPi. The addition of actinomycin D, alpha-amanitin, or rifampin did not inhibit the polymerase activity. After thermal shock of the virus, at least eight different RNA species were synthesized which may correspond to independent transcripts. Transcription also requires a hydrolyzable form of ATP. Analogs such as beta,gamma-imido ATP or beta,gamma-methylene ATP were inhibitory, whereas others, such as the beta-gamma-imido or methylene analogs of CTP, UTP, or GTP, were not inhibitory. This suggests that ATP is related to reactions other than polymerization, probably to initiation or elongation of RNA molecules, as has been described for vesicular stomatitis virus or vaccinia virus. PMID- 6270367 TI - Sequence complexity of circular Epstein-Bar virus DNA in transformed cells. AB - A simplified procedure, based on several methods previously used to isolate circular DNA molecules from bacteria, was derived for the preparation of covalently closed circular viral DNA molecules from large quantities of lymphocytes transformed by Epstein-Barr virus. The protocol can be applied both to virus nonproducer lines and to lines containing cells activated to virus production. Sufficient amounts o highly purified viral DNA of intracellular origin were obtained from B95-8 and Raji cells to allow direct visual analysis of their sequence complexities after cleavage with EcoRI and separation of fragments by gel electrophoresis. No major differences in complexity were observed between circular DNA and linear virion DNA from B95-8 cells. The fragment patterns observed in this fashion agree well with those detected by conventional blotting and hybridization methods. The procedure can also be used as an analytical method to assay for small amounts of circular Epstein-Barr virus DNA molecules in other transformed cells. In this connection, no circular Epstein-Barr virus DNA was detected in Namalva cells. PMID- 6270366 TI - In vitro translation of avian myeloblastosis virus RNA. AB - Avian myeloblastosis virus (AMV)-infected cells contain two viral mRNA's, a genome-sized 34S (7.5-kilobase) mRNA and a 21S (2.5-kilobase) subgenomic mRNA, which contains the AMV-specific sequences (myb sequences). We found that AMV virions packaged both the 7.5-kilobase full-length genomic RNA and the 2.5 kilobase subgenomic RNA. In vitro translation of AMV virion RNA sized by sucrose density gradient centrifugation yielded 76,000-, 56,000-, 48,500-, 47,000-, and 32,000-dalton products. The 76,000-dalton protein was coded for by RNA throughout the gradient, but the peak of activity was at 34S to 35S. The 56,000-, and 48,500 , and 32,000-dalton proteins were encoded in a 21S RNA, and 47,000-dalton protein was encoded in an RNA of approximately 24S. The 76,000-dalton protein was identified as Pr76gag, based upon immunoprecipitation with specific antiserum and the presence of the 19* dipeptide. 7-Methylguanosine triphosphate inhibited the syntheses of Pr76gag and the 56,000-, 48,500-, and 32,000-dalton proteins, but not the synthesis of the 47,000-dalton protein. The 56,000-, 48,500-, 47,000-, and 32,000-dalton proteins were not immunoprecipitated by anti-gag, anti-reverse transcriptase, or anti-gp85 antiserum. Two-dimensional peptide maps of the 56,000 and 48,500-dalton proteins indicated that they were unique. In vitro translational products of myeloblastosis-associated virus 1 were also analyzed to aid in the identification of the AMV myb gene product(s); the translational products analyzed included Pr76gag, p60env, and a 56,000-dalton polypeptide which apparently was not identical to the 56,000-dalton AMV translational product, as determined by two-dimensional peptide mapping. Our data indicated that one of these proteins (56,000, 48,500, or 32,000 daltons) may represent the product of the AMV myb gene and, therefore, the putative transforming protein(s) of AMV. PMID- 6270369 TI - Identification of a virus-specific polypeptide associated with a transforming fragment (BglII-N) of herpes simplex virus type 2 DNA. AB - The BglII N fragment of herpes simplex virus type 2 (HSV-2) DNA (approximately 0.58 to 0.63 map unit) was examined for encoded products. Using plasmid pGZ59, which consists of BglII-N cloned in pAT153, in conjunction with hybrid arrested translation, mRNA selection, and in vitro protein synthesis, we found that the major translated product of this region has an approximate molecular weight of 37,800. By further mapping, coding sequences for this polypeptide were located within the region of BglII-N representing approximately 0.58 to 0.61 genome map unit. To demonstrate immunological specificity, we used staphylococcal A protein immunoprecipitation with rabbit anti-HSV-1 or HSV-2 sera and antigens from HSV-1 or HSV-2 total mRNA translated in vitro and BglII-N-selected mRNA. The results show that the 37,800-dalton polypeptide has HSV-2 immunological specificity, as it is precipitated with anti-HSV-2 sera but not with anti-HSV-1 or control sera. PMID- 6270368 TI - Restriction map of the single-stranded DNA genome of Kilham rat virus strains 171, a nondefective parvovirus. AB - We constructed a physical map of Kilham rat virus strains 171 DNA by analyzing the sizes and locations of restriction endonuclease-generated fragments of the replicative-form viral DNA synthesized in vitro. BglI, KpnI, BamHI, SmaI, XhoI, and XorII did not appear to have any cleavage sites, whereas 11 other enzymes cleaved the genome at one to eight sites, and AluI generated more than 12 distinct fragments. The 30 restriction sites that were mapped were distributed randomly in the viral genome. A comparison of the restriction fragments of in vivo- and in vitro-replicated replicative-form DNAs showed that these DNAs were identical except in the size or configuration of the terminal fragments. PMID- 6270370 TI - Characterization of bacteriophage N3 DNA. AB - The DNA from Haemophilus influenzae temperate phage N3 was characterized by centrifugation and by electrophoresis after nuclease digestion. The double stranded DNA, with a mass of 25.8 X 10(6) daltons, had single-strand cohesive ends. Strand association through cohesion was reduced by heat and removed by S1 nuclease digestion. N3 DNA contained five EcoRI, one KpnI, two SacI, six XbaI, and four XhoI cleavage sites. The cohesive end segments were identified by heating the digests before electrophoresis. This was the first step in the construction of the physical maps of this DNA. PMID- 6270371 TI - DNA synthesis and template activity in a mutant of polyoma virus with altered expression of late viral proteins. AB - The 3049 strain of polyoma virus overproduces late RNA and proteins. The synthesis and accumulation of virus DNA was measured in cells infected with 3049 and a wild-type virus, lpS, to assess the possible role of gene dosage in this phenomenon. The rate of incorporation of [3H]thymidine into the DNA of 3049 and lpS virus was shown to be identical. The number of genome copies per cell, determined by DNA-DNA reassociation kinetics, was found to be similar in either whole cell or Hirt extracts of cells infected with the two viruses. Viral transcription complexes were isolated by Sarkosyl extraction, and the specific transcriptional activity was measured by incorporation of [3H]UTP. The incorporation of [3H]UTP and the DNA content of transcription complexes were indistinguishable. These results suggest that overproduction of late RNA (and protein) by the 3049 virus is not due to either an increased number of viral genomes per cell or an increased fraction of genomes involved in transcription. These and other data (unpublished) support the conclusion that the altered phenotype is due to a posttranscriptional mechanism. PMID- 6270372 TI - Viral gene expression in polyoma virus-transformed rat cells and their cured revertants. AB - We have studied transcription of integrated viral DNA sequences in a variety of ts-a polyoma virus-transformed rat cells and cured revertants (which had undergone excision of variables amounts of integrated viral DNA) to characterize the structure of viral mRNA's produced in these lines under conditions in which integrated DNA is stable. Our results indicate that cells containing intact early region sequences, either in single-copy or tandem insertions, produce mRNA's indistinguishable from those observed early in lytic infections; sequences complementary to the polyoma late region were not transcribed from integrated viral DNA. Cured revertants no longer encoded full-length early mRNA's , but produced viral transcripts whose 3' ends mapped at an alternative early region polyadenylic acid attachment site at 99 map units or extended in to flanking host sequences. The phenotype of these revertant cells correlated with the abundance of these transcripts, suggesting that the transforming function(s) of polyoma virus controls the cellular phenotype in a dose-dependent manner. Unexpected results were obtained from studies of cells containing tandem repeats of defective viral DNA in which the polyadenylic acid attachment signal at 25.8 map units and surrounding sequences were deleted. In these cases, polyadenylated mRNA's were observed that contained sequences complementary to the early strand of the polyoma late region. These mRNA's (some larger than 8 kilobases) originated at the viral early promoter, extended into the late region, and continued into the early region of the contiguous repeat in the tandem. The multimeric mRNA's produced contained defective early regions in tandem with late region sequences. S1 analysis indicated that whereas the 5' early region sequences of readthrough transcripts were spliced in the usual manner, internal early region repeats were either unspliced or used only one of the small early region splices. When deletions in the viral readthrough transcripts were observed. This suggests that sequences nearby the AAUAAA sequence at 26 map units may control transcription termination of the polyoma early region. PMID- 6270373 TI - Differences between poliovirus empty capsids formed in vivo and those formed in vitro: a role for the morphopoietic factor. AB - Empty capsid species formed from the self- and extract-mediated assembly of poliovirus type 1 14S particles in vitro and procapsids isolated from virus infected cells were subjected to isoelectric focusing in charge-free agarose gels. The empty capsid formed in the self-assembly reaction had an isoelectric point (pI) of 5.0, whereas procapsids and extract-assembled empty capsids focused at pH 6.8. Unreacted 14S particles focused at pH 4.8 to 5.0. The sedimentation coefficient (s20,w) and density of the empty capsid species were also determined. Procapsids had a density in CsCl of 1.31 g/cm3, whereas empty capsids formed by self- or extract-mediated assembly had a density of 1.29 g/cm3. Both extract assembled empty capsids and procapsids had an s20,w of 75S, whereas self assembled empty capsids had an s20,w of 71S. Self-assembled empty capsids were not converted to pI 6.8 empty capsids by incubation with poliovirus-infected HeLa cell extracts. The dissociated polypeptides of self-assembled empty capsids (pI 5.0) and procapsids (pI 6.8) behaved identically when analyzed by isoelectric focusing in the presence of 9 M urea and by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. These results suggest that infected cell extracts possess a factor that influences the final conformation of the empty shell (pI 6.8, 75S) formed from 14S particles and that this influences is exerted at the initiation step or during the polymerization reaction. A small amount of this activity (less than or equal to 20% of infected extracts) was detected in uninfected cells; the significance of this remains unknown. PMID- 6270374 TI - Evidence that the UV endonuclease activity induced by bacteriophage T4 contains both pyrimidine dimer-DNA glycosylase and apyrimidinic/apurinic endonuclease activities in the enzyme molecule. AB - We performed experiments to determine whether the phage T4-induced UV endonuclease activity is a single protein containing both pyrimidine dimer-DNA glycosylase and apyrimidinic endonuclease activities. The UV endonuclease activity is induced by the denV gene and codes for the glycosylase activity. We obtained several kinds of evidence that the protein containing the glycosylase activity also contains the apyrimidinic endonuclease activity. After chromatography on DEAE-cellulose, the two activities copurified during phosphocellulose chromatography and Sephadex G-100 chromatography, with a constant ratio of activities across the activity peaks. On Sephadex G-100 columns the molecular weights of the two activities agreed within 2,500 or less. When an extract of cells infected with the T4 V1 mutant was purified in exactly the same way as an extract of cells infected with T4 V1+, neither glycosylase nor apyrimidinic endonuclease activity was detected in the normal elution position of the T4 UV endonuclease activity. The glycosylase and apyrimidinic endonuclease activities were induced with similar kinetics, which were characteristic of immediate early rather than delayed early enzymes. This correlated well with the presumed major role of these activities in repairing thymine dimers in parental DNA before DNA replication begins. Finally, glycosylase and apyrimidinic endonuclease activities were lost in parallel during incubation of the enzyme at 46 degree C. Our results indicated that both of these enzyme activities are contained in the same enzyme molecule and, probably, in the same polypeptide. PMID- 6270375 TI - den V gene of bacteriophage T4 codes for both pyrimidine dimer-DNA glycosylase and apyrimidinic endonuclease activities. AB - Recent studies have shown purified preparations of phage T4 UV DNA-incising activity (T4 UV endonuclease or endonuclease V of phage T4) contain a pyrimidine dimer-DNA glycosylase activity that catalyzes hydrolysis of the 5' glycosyl bond of dimerized pyrimidines in UV-irradiated DNA. Such enzyme preparations have also been shown to catalyze the hydrolysis of phosphodiester bonds in UV-irradiated DNA at a neutral pH, presumably reflecting the action of an apurinic/apyrimidinic endonuclease at the apyrimidinic sites created by the pyrimidine dimer-DNA glycosylase. In this study we found that preparations of T4 UV DNA-incising activity contained apurinic/apyrimidinic endonuclease activity that nicked depurinated form I simian virus 40 DNA. Apurinic/apyrimidinic endonuclease activity was also found in extracts of Escherichia coli infected with T4 denV+ phage. Extracts of cells infected with T4 denV mutants contained significantly lower levels of apurinic/apyrimidinic endonuclease activity; these levels were no greater than the levels present in extracts of uninfected cells. Furthermore, the addition of DNA containing apurinic or apyrimidinic sites to reactions containing UV-irradiated DNA and T4 enzyme resulted in competition for pyrimidine dimer-DNA glycosylase activity against the UV-irradiated DNA. On the basis of these results, we concluded that apurinic/apyrimidinic endonuclease activity is encoded by the denV gene of phage T4, the same gene that codes for pyrimidine dimer-DNA glycosylase activity. PMID- 6270376 TI - Simian virus 40 early mRNA's contain multiple 5' termini upstream and downstream from a Hogness-Goldberg sequence; a shift in 5' termini during the lytic cycle is mediated by large T antigen. AB - We have used primer-directed synthesis, separation, and sequencing of cDNA's to identify and localize the 5' termini of simian virus 40 early mRNA's. We have examined polyadenylated RNAs obtained from whole cytoplasm and polysomes of two transformed lines and from the cytoplasm of infected cells early and late in the lytic cycle, and we have attempted to correlate the results of our cDNA analyses with recent analyses of early cap structures. We have found that early mRNA's from transformed cells have three principal 5' termini, at residues 5,150, 5,154, and 5,155, with terminal transcribed sequences of CU, GC, and GG, respectively. These termini lie 21 to 26 nucleotides downstream from the early Hogness-Goldberg sequence. Transformed cell early mRNA's also contain a series of less abundant 5' termini that are copied from DNA sequences as far as 80 nucleotides downstream and a minimum of 70 to 75 nucleotides upstream from the Hogness-Goldberg sequence. The templates for the upstream 5' termini and the late simian virus 40 mRNA's overlap by a minimum of 60 to 65 nucleotides. Early mRNA's isolated from cells early in infection contain the same three principal 5' termini and downstream minor 5' termini as transformed cell mRNA's, but they lack 5' termini upstream from the Hogness-Goldberg sequence. With the onset of the late lytic phase, there is a progressive decreases in the utilization of the three principal 5' termini and additional downstream 5' termini and a progressive increase in the utilization of four major termini at residues 5,190 to 5,194, which are 10 to 15 nucleotides upstream from the Hogness-Goldberg sequence. With the onset of the late lytic phase, there is a progressive decrease in the utilization of the three principal 5' termini and additional downstream 5' termini and a progressive increase in the utilization of four major termini at residues 5,190 to 5,194, which are 10 to 15 nucleotides upstream from the Hogness-Goldberg sequence. This shift is evident in cells infected with a tsA mutant at the permissive temperature, but is aborted by growth at or shift-up to a restrictive temperature. Thus, this shift is mediated by the gene A product, large T antigen. We present two models, which are mutually exclusive, to account for the role of T antigen in the early-late shift. One involves transcription late in infection on a new DNA template synthesized during DNA replication. The second involves inhibition of initiation of early transcription at residues 5,150 to 5,155 and other downstream sites and a shift of transcription initiation principally to the upstream sites as a result of the binding of T antigen to two sites on simian virus 40 DNA downstream from the Hogness-Goldberg sequence. PMID- 6270377 TI - Herpes simplex virus types 1 and 2 completely help adenovirus-associated virus replication. AB - In addition to adenoviruses, which are capable of completely helping adenovirus associated virus (AAV) multiplication, only herpesviruses are known to provide any AAV helper activity, but this activity has been thought to be partial (i.e., AAV DNA, RNA, and protein syntheses are induced, but infectious particles are not assembled). In this study, however, we show that herpes simplex virus type 1 (HSV 1) and type 2 (HSV-2) are in fact complete AAV helpers and that AAV type 2 (AAV2) infectivity yields can approach those obtained when coinfections are carried out with a helper adenovirus. AAV helper activity was demonstrated in KB cells with two HSV-1 strains (11124 and 17MP) and an HSV-2 strain (HG52). Each herpesvirus supported AAV2 multiplication with comparable efficiency. AAV2 multiplication was similarly efficient in HSV-1 coinfections of HeLa cells, whereas lower yields were obtained in HEp-2 and primary human embryonic kidney cells. HSV-1 also supported AAV1 multiplication in HeLa cells but, at corresponding multiplicities of infection, AAV1 grew less efficiently than AAV2. Comparisons of the time courses of AAV2 DNA, RNA, and protein syntheses after coinfection with either adenovirus type 5 or HSV-1 revealed that, in each case, the onset of synthesis and attainment of maximal synthesis rate occurred earlier in coinfections with HSV-1. These findings demonstrate the linkage of AAV macromolecular synthesis to an event(s) in the helper virus cycle. Aside from this temporal association, helper-related differences in AAV macromolecular synthesis were not apparent. PMID- 6270378 TI - Genetic structure, transforming sequence, and gene product of avian sarcoma virus UR1. AB - We analyzed the genetic structure and gene products of the newly isolated avian sarcoma virus UR1, which recently has been shown to be replication defective and to contain no sequences homologous to the src gene of Rous sarcoma virus. The sizes of the genomic RNAs of UR1 and its associated helper virus, UR1AV, were determined to be 29S and 35S (5.9 and 8.5 kilobases), respectively, by gel electrophoresis and sucrose gradient sedimentation. RNase T1 oligonucleotide mapping of purified viral RNAs indicated that UR1 RNA contains eight unique oligonucleotides in the middle of the genome and shares four 5'-terminal and three 3'-terminal oligonucleotides with UR1AV RNA. The unique sequences of UR1 and Fujinami sarcoma virus were found to be closely related to each other by molecular hybridization of UR1 RNA with DNA complementary to the unique sequence of Fujinami sarcoma virus RNA, but minor differences were found by oligonucleotides fingerprinting. In the regions flanking the unique sequences, UR1 and Fujinami sarcoma viral RNAs contain distinct oligonucleotides, which are shared with oligonucleotides of the respective helper viral RNAs. Cell transformed with UR1 produce a single 29S RNA species which contains a UR1 unique sequence; this species is most likely the mRNA coding for the transforming protein. In UR1-transformed cells, a phosphoprotein fo 150,000 daltons (p150) was detected by immunoprecipitation with antiserum against gag proteins. p150 was associated with a protein kinase activity that was capable of phosphorylating p150 itself, immunoglobulin G of antiserum, and a soluble substrate, alpha casein. This enzyme transferred phosphate exclusively to tyrosine residues of substrates in vitro, but p 150 labeled in vivo with 32P contained both phosphoserine and phosphotyrosine. The in vitro kinase reaction was not affected by the presence of cyclic AMP or cyclic GMP and strongly preferred Mn2+ over Mg2+. Thus, the properties of UR1 protein are almost identical to those of Fujinami sarcoma virus protein. PMID- 6270379 TI - Some biological properties of two new avian sarcoma viruses. AB - The new avian retroviruses UR1 and UR2 were isolated from spontaneous tumors of chickens by cocultivation of tumor material with susceptible chicken embryo fibroblasts. In vitro, UR1 induced formation of small foci of round and fusiform cells. On the other hand, cells infected by UR2 assumed an extremely elongated morphology. In vivo, both viruses induced fibrosarcomas and myxosarcomas with short latencies. Infectivity assays with and without mitomycin C showed that both viruses were defective for replication, but transformed nonproducing cell clones were obtained only with UR1. UR1-infected transformed nonproducing clones did not release particles detectable by reverse transcriptase assays, and fusion of transformed nonproducing cells with quail cells chronically infected with Rous sarcoma virus (a Bryan strain) failed to rescue infectious virus. This suggested that UR1 does not code for functional envelope glycoproteins. In this regard, UR1 appeared to be similar to Fujinami, PRCII, and Y73 viruses. The helper viruses of partially purified stocks of UR1 and UR2 appeared to belong to subgroup A, but these helper viruses were distinguishable from each other, as shown by host range experiments and neutralization tests. Hybridization studies with DNA complementary to the src gene of Rous sarcoma virus and RNAs extracted from both UR1 and UR2 showed no homology between the genomes of the new isolates and the transforming gene of Rous sarcoma virus. PMID- 6270380 TI - BK virus-transformed inbred hamster brain cells. I. Status of the viral DNA and the association of BK virus early antigens with purified plasma membranes. AB - Inbred LSH hamster brain cells were transformed in vitro by the GS strain of BK virus (BKV), and transplantable tumors classified as undifferentiated glioblastomas were induced in the syngeneic host. The viral status in the transformed cells, designated LSH-BR-BK, was established. About 46 genome equivalents per cell of viral DNA was detected, with the majority of sequences in a free form. The transformed cells expressed large quantities of tumor (T) antigen as well as surface (S) antigen as demonstrated by indirect immunofluorescence. Sixty-three percent of tumor-bearing hamsters produced high titer antibodies against T, whereas 3 of 14 (21%) hamsters also produced antibodies against the BKV-specific S antigen. Furthermore, the relatedness of BKV early gene products, including T, S, and tumor-specific transplantation antigen, was established by the production of a rabbit antiserum against highly purified plasma membranes of LSH-BR-BK cells and by the induction of a BKV specific tumor-specific transplantation antigen response by these plasma membranes in the syngeneic host. PMID- 6270382 TI - Focus formation and neoplastic transformation by herpes simplex virus type 2 inactivated intracellularly by 5-bromo-2'-deoxyuridine and near UV light. AB - The induction of focus formation in low serum and of neoplastic transformation of Syrian hamster embryo cells was examined after the expression of herpes simplex virus type 2 functions. Syrian hamster embryo cells infected at a high multiplicity (5 PFU/cell) with 5-bromo-2'-deoxyuridine-labeled herpes simplex virus type 2 (11% substitution of thymidine residues) were exposed to near UV light irradiation at various times postinfection. This procedure specifically inactivated the viral genome, while having little, if any, effect on the unlabeled cellular DNA. Focus formation in 1% serum and neoplastic transformation were observed in cells exposed to virus inactivated before infection, but the frequency was enhanced (15- to 27-fold) in cells in which the virus was inactivated at 4 to 8 h postinfection. Only 2 to 45 independently isolated foci were capable of establishing tumorigenic lines. The established lines exhibited phenotypic alterations characteristic of a transformed state, including reduced serum requirement, anchorage-independent growth, and tumorigenicity. They retained viral DNA sequences and, even at relatively late passage, expressed viral antigens, including ICP 10. PMID- 6270381 TI - Phosphorylation patterns of tumour antigens in cells lytically infected or transformed by simian virus 40. AB - The phosphorylation sites of simian virus 40 (SV40) large tumor (T) antigens have been analyzed by partial proteolysis peptide mapping and phosphoamino acid analysis of the resulting products. At least four sites were found to be phosphorylated. An amino-terminal part of the molecule contained both phosphoserine and phosphothreonine. One phosphothreonine residue was located in the proline-rich carboxy-terminal end of the molecule, either at position 701 or at position 708. The mutant dl 1265, which is defective in adenovirus helper function, lacked this phosphorylation site. In addition, the carboxy-terminal part of the molecule contained phosphoserine at a more central position. T antigen-associated proteins of SV40-transformed cell (nonviral T; 51,000 to 55,000 daltons) also contained multiple phosphorylation sites involving at least two serine residues in mouse antigens and an additional threonine residue in rat, human, and monkey antigens. The latter residue and at least one phosphoserine residue were located near one terminus of the human NVT molecule. We did not find any evidence for phosphorylation of tyrosine residues in any of the multiple species of either large T or nonviral T molecules. Several forms of large T antigens were extracted from both SV40-transformed and SV40-infected permissive and nonpermissive cells, and their phosphorylation patterns were compared. No evidence was found for a different phosphorylation pattern of T antigen in transformed cells. PMID- 6270383 TI - Two virus-specific rna species are present in cells transformed by defective leukemia virus OK10. AB - OK10 is a defective leukemia virus which shares some biological and biochemical properties of avian myelocytomatosis virus (MC29). We investigated the pattern of transcription of OK10 in both quail and chicken cells. In both cell types, OK10 produced two polyadenylated RNA species of 8.6 and 3.5 kilobases, which both contained sequences derived from the 5' end of the genome and also the presumed transforming gene (myc). This is a novel form of expression for defective leukemia viruses of the MC29 subgroup and may indicate that there is an as-yet unidentified protein produced in OK10-infected cells which may be involved in transformation. PMID- 6270384 TI - Varicella-zoster virus vaccine DNA differs from the parental virus DNA. AB - The DNAs of a varicella-zoster virus vaccine and its parental virus were compared by CsCl buoyant density centrifugation and restriction enzyme cleavage analysis. The varicella-zoster virus vaccine DNA showed a heterogeneous buoyant profile and altered restriction enzyme cleavage patterns. These changed properties are probably the result of the accumulation of virus containing defective varicella zoster virus DNA during extensive cell culture passage of the vaccine virus. PMID- 6270385 TI - Regulation of herpesvirus macromolecular synthesis: temporal order of transcription of alpha genes is not dependent on the stringency of inhibition of protein synthesis. AB - Operationally, alpha genes of herpes simplex virus 1 were defined on the basis of the observations that they are the earliest genes expressed in the infected cell and that the transcription, processing, accumulation of the mRNA's in the infected cell cytoplasm can take place in the presence of inhibitors of protein synthesis, such as cycloheximide. In these studies, we translated in vitro the viral mRNA's extracted from cells infected maintained in the presence of cycloheximide, emetine, or anisomycin. Inasmuch as all the major alpha proteins (no. 0, 4, 22, and 27) were translated, we conclude that the transcription of all previously defined alpha genes is independent of the stringency of inhibition of protein synthesis and that pre-alpha genes cannot be detected in such experiments. PMID- 6270386 TI - Aberrant glycoprotein mRNA synthesized by the internal deletion mutant of vesicular stomatitis virus. AB - The internal deletion mutant (DI-LT) derived from the heat-resistant strain of vesicular stomatitis virus synthesized an aberrant polyadenylated mRNA in vivo and in vitro. No normal glycoprotein message could be detected among the in vivo transcription products. The abnormal RNA contained a transcript of the partially deleted polymerase gene covalently linked to the 3' end of the glycoprotein message. The polyadenylate is located at the 3' end of the molecule and is most probably encoded by the remnant polymerase gene polyadenylation signal. This aberrant RNA may be synthesized because of either a failure to terminate transcription at the end of the glycoprotein gene or an inability to process an abnormal polycistronic precursor. PMID- 6270387 TI - Quantitative of murine mammary tumor virus-related RNA in mammary tissues of low- and high-mammary-tumor-incidence mouse strains. AB - Lactating mammary glands and hormonally induced mammary tumors of BALB/c mice from three geographically separated breeding colonies were examined by molecular hybridization, using murine mammary tumor virus (MuMTV) cDNA representing the entire viral genome to determine the amount of MuMTV-related RNA expressed in these tissues. The RNA extracted from these tissues by the classical sodium dodecyl sulfate-pronase, phenol-chloroform procedure (method 1) contained barely detectable levels of MuMTV-related sequences. In contrast, both normal lactating mammary glands and hormonally induced mammary tumors of these mice were found to contain approximately one to two copies of the MuMTV genome per cell by using a new procedure in which the RNA was extracted with guanidine derivatives (method 2). No significant differences in the MuMTV-related RNA content of the BALB/c mammary tissues were observed regardless of their colony of origin. Our results suggest that expression of MuMTV RNA does not change in BALB/c mammary glands during transformation to a malignant state and that MuMTV expression does not play a role in tumorigenesis in these mice. In view of the increased recovery of MuMTV-related RNA from BALB/c mice with method 2, we compared the level of MuMTV RNA expression in lactating mammary glands and mammary tumors of other mouse strains, including C57BL/6 and RIII, using both extraction methods. Yields of MuMTV-related RNA from mammary tissues increased by as much as 35- to 40-fold, using method 2 as compared with method 1. Therefore method 2, involving guanidine derivatives, appears to be method of choice for MuMTV-related RNA extraction from the mammary tissues of certain strains of mice, particularly those expressing relatively low levels of MuMTV RNA. PMID- 6270389 TI - Congenital mesoblastic nephroma: relationship to other renal tumors of infancy. AB - Although nephroblastomas (Wilms tumor) do occur in the newborn most of the solid renal tumors in this age group constitute a distinctly different pathologic entity, congenital mesoblastic nephroma. While a more cellular histologic variant of congenital mesoblastic nephroma has been recognized recently, which occasionally may follow a malignant course, the tumor with conventional histology has never been reported to metastasize and follows a benign course when treated by adequate local excision alone. We report our 50-year experience with 11 cases of this tumor (9 cases with conventional histology and 2 with cellular variant histology). Typically, the tumor presented in the newborn as a large asymptomatic mass. Local invasion was never seen and the tumors were resected without difficulty. Grossly, the tumors usually resembled a uterine "fibroid" and, unlike nephroblastomas, rarely exhibited hemorrhage or necrosis. Microscopically, the conventional histologic congenital mesoblastic nephroma was composed of sheets of spindle-shaped cells. The cellular variant of congenital mesoblastic nephroma exhibited increased cellularity and a higher mitotic index. The relationship of congenital mesoblastic nephromas to other neonatal renal tumors is discussed and a unifying schema of neoplasia in infantile renal tumors is proposed. PMID- 6270388 TI - Endogenous type C retroviral sequences of mice are organized in a small number of virus-like classes and have been acquired recently. AB - We studied endogenous type C virus-related sequences of mice by annealing Moloney murine leukemia virus DNA to agarose gel blot transfers of uninfected mouse cell DNA which had been cleaved with restriction enzymes. We found that many of the endogenous murine leukemia virus-related sequences in mice consist of two organizational classes that are integrated into many different loci. Both of these classes resemble standard murine leukemia virus proviral DNA in both size and sequence organization. All lines of inbred mice examined contained both organizational classes, as did feral isolates of Mus musculus domesticus. However, a related Asian subspecies, Mus musculus molossinus, contained different organizational classes of endogenous murine leukemia virus-related sequences. Among inbred strains and feral isolates of M. musculus domesticus, the murine leukemia virus-related sequences were present at different loci. This suggested that most of these sequences were acquired relatively recently during subspeciation and inbreeding. PMID- 6270390 TI - Carcinoembryonic antigen. A useful monitor of therapy of small cell lung cancer. AB - In 85 patients with small cell cancer of the lung, the degree of carcinoembryonic antigen (CEA) elevation correlated with the extent of disease. Normal CEA levels (less than 2.5 ng/mL) occurred in 55% of patients with limited disease compared with 12% of patients with extensive disease. All patients with CEA levels greater than 50 ng/mL had liver involvement. Serial CEA levels were useful in monitoring response to chemotherapy and in predicting relapse. All patients with CEA values greater than 5.0 ng/mL before start of treatment had a fall in CEA level with response to treatment and a rise in CEA level when resistance to chemotherapy developed. In patients with CEA levels of less than 5.0 ng/mL at the start of treatment, rising values reflected the onset of recurrent disease in 10 or 21 patients who had relapses. The CEA levels used in conjunction with standard roentgenograms and organ scanning studies can aid in the initial staging of small cell cancer of the lung, and in monitoring treatment. PMID- 6270391 TI - [Antibacterial activity of cefotaxime against main bacteria isolated from clinical samples (author's transl)]. PMID- 6270392 TI - [Studies on cefotaxime (HR 756) concentration in human biliary tract and clinical effect for acute or subacute cholecystitis with cholelithiasis (author's transl)]. AB - A new antibiotic drug of cephalosporin with marked resistance to beta-lactamase, cefotaxime (HR 756) for parenteral use in 8 patients with acute or subacute cholecystitis with cholelithiasis. Cefotaxime was administrated by intravenous injection or drip infusion at a daily dose of 1-4 g. Clinical response was excellent in 1 case, good in 7 cases, and fair or poor was none. Clinical adverse effect was not recognized. Cefotaxime in a dose of 1 g was given intravenously during operation to those same 8 patients. Tissue specimens of different places were taken from removed organs. The materials of A-bile and B-bile were subsequently taken at intervals. Determination of cefotaxime concentration was performed according to the bioassay method with Micrococcus luteus ATCC 9341 strain. Cefotaxime concentrations in the A-bile increased gradually until 1 hour after the intravenous administration. Cefotaxime was observed in the B-bile through the gallbladder wall after the intravenous injection. PMID- 6270393 TI - [Reproduction studies on ceftizoxime sodium in rats (author's transl)]. PMID- 6270394 TI - [Clinical studies on cefotaxime (CTX, HR 756) in obstetrics and gynecology (author's transl)]. AB - In vivo transfer and therapeutic efficacy of a new cephalosporin derivative, cefotaxime, which is stable against beta-lactamase hydrolysis, have studied in gynecology field. The following results have been obtained. (1) The level of cefotaxime transferred to uterus artery and to uterus was higher than its MIC against majority of Gram-negative bacilli, such as E. coli. (2) Transfer of this drug to umbilical blood was also satisfactory. (3) This drug as demonstrated its efficacy in treating 8 infection cases refractory to CET, CEZ and ABPC, out of which 3 had 'excellent' and 5 had 'good' results. (4) No side effect was evidenced in any of our patients. In conclusion, this drug has satisfactory tissue transfer as well as sufficient safety and excellent efficacy in treatment of gynecological infection cases. PMID- 6270395 TI - [Absorption and excretion of cefotaxime and its levels in uterine arterial blood, female internal genital organ tissue and pelvic cavity fluid (author's transl)]. AB - Following results were obtained from intravenous single administration of cefotaxime 2 g by measuring its levels in uterine arterial blood, elbow venous blood, ovary, oviduct, several sites in uterine tissue, and in the fluid excreted in the dead space formed by radical hysterectomy. (1) Uterine artery and elbow vein blood levels revealed marked increase immediately after administration followed by gradual reduction at slow rate. (2) Peak levels in female internal genital organ tissue were achieved 12 approximately 19 minutes after cefotaxime administration, and ranged between 5 to 8 mcg/g independently from measurement site. In cervix uteri, portio vaginalis high concentration of 4.9 approximately 5.7 mcg/g was achieved within ten minutes after drug administration. (3) In the fluid excreted in the dead space formed by radical hysterectomy cefotaxime levels were maintained around 15 mcg/ml level throughout 5 approximately 6 hours following its administration, and gradually decreased at slow rate later. PMID- 6270396 TI - [Clinical studies of cefotaxime (CTX, HR 756) for infectious diseases in the field of obstetrics and gynecology (author's transl)]. AB - Ten cases of gynecological infection were treated successfully by a new cephalosporin derivative, cefotaxime (HR 756, CTX), which proved excellent in 4 cases and good in 6 cases. No adverse reaction was found to be induced by administration of this drug. Post-administration results were all normal for both renal and hepatic function tests. It is extremely difficult to accurately assess bacteriological efficacy of a drug in treatment of female genital infections. Nevertheless, we determined its MICs for a few number of strains we could isolate from clinical cases. Its MIC against 4 strains of B. fragilis inoculated at 10(8) cells/ml was 25 approximately 50 mcg/ml and was 1.56 approximately 3.13 mcg/ml at 10(6) cells/ml inoculation. Its MIC against 2 strains of Ps. cepacia was 12.5 mcg/ml at 10(8) cells/ml inoculation and 6.25 mcg/ml at 10(6) cells/ml inoculation. These MICs suggest that this drug is a very promising antimicrobial agent for treatment of gynecological infections, as been already indicated by clinical results of this drug. PMID- 6270397 TI - [Laboratory and clinical studies on cefotaxime in obstetrics and gynecology (author's transl)]. AB - The following results have been obtained in our basic and clinical studies to examine a new cephalosporin derivative, cefotaxime (CTX). For basic study purpose, we injected 1 g of CTX to 6 patients who had received simple total hysterectomy and measured its level in elbow vein, uterine artery, corpus uteri, cervix uteri, endometrium, oviduct and ovary. Any definitive conclusion should not be drawn from these results, since they have too big variation. However, slightly higher levels were achieved cervix uteri and oviduct than in other organs in a same patient. For clinical evaluation, we tried this drug to 12 cases of female genital infections and 91.7% response rate was obtained. Our studied population includes 8 cases of adnexitis, 2 with pelvioperitonitis, 1 with panperitonitis, and 1 patient with BARTHOLIN abscess. Organisms were detected only in 3 cases, including S. aureus in one patient, Micrococcus sp. in one, and mixture of Klebsiella sp. and B. fragilis in 1. Puncture of DOUGLAS pouch was performed in 3 cases after the completion of therapy and all had negative results. No side effect was observed, and no significant difference was noted between pre- and post-therapy examinations of peripheral blood, hepatic function and renal function. PMID- 6270398 TI - [Clinical studies of cefotaxime, administration in intravenous drip infusion, for infectious diseases in the field of obstetrics and gynecology (author's transl)]. AB - Therapeutic efficacy and safety of a new cephalosporin for injection, cefotaxime (CTX) have been examined in gynecological infection cases. CTX has demonstrated in this study excellent therapeutic efficacy in infections with aerobes only, anaerobes only and with mixture of aerobes and anaerobes. No side effect has been observed in our patients who had been administered total dose of 8 approximately 28 g during the period of 4 approximately 7 days. PMID- 6270399 TI - [Bacteriological and clinical studies on cefotaxime in obstetrics and gynecology (author's transl)]. AB - Fundamental and clinical studies of cefotaxime (CTX), a new semisynthetic cephalosporin antibiotic were carried out in the field of obstetrics and gynecology. The following results were obtained. 1. CTX was almost equally active to SCE-1365 and less active than cefotiam (CTM) and cefazolin (CEZ) against S. aureus, much more active than these 3 antibiotics against E. coli, more active than the 3 antibiotics against Enterobacter, equally active to SCE-1365 and a little weaker than CEZ against anaerobic Gram positive cocci, and superior to SCE 1365 and CEZ against Bacteroides. 2. CTX was more stable to B. fragilis-producing beta-lactamase than CET, CEZ and cefoperazone (CPZ). 3. The concentrations of CTX transferred to the female genital organs after CTX 1 g d. i. were sufficiently effective against facultative or strict anaerobic bacteria mainly isolated from obstetrical and gynecological infections. 4. CTX was administered to 10 patients with genital infections. CTX was excellent in 1 case, good in 8 cases and poor in 1 case. The response rate of CTX was 90%. Bacteria, isolated in 5 cases before CTX treatment, were eradicated in 4 cases. The bacteriological effectiveness of CTX was thus 80%. 5. No side effect attributable to CTX was observed. PMID- 6270400 TI - [Fundamental and clinical studies on cefotaxime in the field of obstetrics and gynecology (author's transl)]. AB - Fundamental and clinical studies were made on cefotaxime (HR-756, CTX), a new cephalosporin antibiotic, with the following results. 1. Cefotaxime was given to 22 patients. Efficacy was excellent in 3 cases, good in 17 cases and poor in 2 cases. No side effects were observed in any cases. 2. Following a single intravenous injection of 1.0 or 2.0 g, the transfer of CTX to the internal genital organs was found to be good. The transfer of CTX to exudate of the dead space of pelvis was also good. 3. The above data demonstrated that CTX is a safe and effective drug. PMID- 6270401 TI - [Fundamental and clinical studies on cefotaxime in the field of obstetrics and gynecology (author's transl)]. AB - Fundamental and clinical studies were made on cefotaxime (CTX), a new cephalosporin antibiotic. The following results were obtained: 1. Antibacterial activity: At a concentration of 3.13 mcg/ml, CTX inhibited the growth of 90.2% of 92 strains of Gram-negative rods and 80.0% of 15 strains of Gram-positive cocci. 2. Concentrations of CTX in body fluids and genital organs after 2 g i.v.: (1) CTX level in pus reached the peak (5.6mcg/ml) at 2 hours after administration. (2) Mean CTX levels in the pelvic space exudate reached the peak (28.0 mcg/ml) at 2 hours after administration. (3) CTX levels in the uterine appendages and uterus reached the peak (8.9 and 4.5 mcg/g, respectively) at 100 to 280 minutes after administration. 3. CLINICAL RESULTS: CTX was excellent in 7 of 13 cases and good in the remaining 6 cases. The response rate to CTX was 100%. 4. The bacteriological effect: The bacteriological effect of CTX was also 100%. Bacteria were eradicated in 7 of the 10 cases where organisms were demonstrated before CTX treatment. Partial reduction bacteria was observed in the remaining 3 cases. 5. No side effect attributable to CTX was observed. PMID- 6270402 TI - [Transfer of cefotaxime to the pelvic organs (author's transl)]. AB - The new antibiotic cefotaxime (HR 756, CTX) has been proved to be clinically effective against infections observed in the field of obstetrics and gynecology. The present study was intended to investigate the transfer of CTX to the internal genital organs and the dead pelvic space. The results were obtained as follows: 1. The concentrations of CTX transferred to the uteri and its appendages after CTX 1 g intravenous injection were sufficiently effective against the major pathogens (Gram-negative and anaerobic bacteria) demonstrated in the field of obstetrics and gynecology. 2. The concentrations of CTX transferred to the dead space of the pelvis were effective against almost all of the Gram-negative bacteria for 6 to 12 hours after CTX 1 g or 2 g intravenous injection. CTX was thus proved to be very effective for the prevention and treatment of infections of the dead pelvic space. PMID- 6270403 TI - [Laboratory and clinical studies on cefotaxime in obstetrics and gynecology (author's transl)]. AB - We have conducted basic and clinical examination of a new cephalosporin derivative, cefotaxime (CTX). The average level of transfer measured at various locations in uterus tissue and adnexa 15 to 105 minutes (53.4 minutes in average) after intravenous administration of this drug ranged from 1.5 mcg/g (myometrium) to 3.3 mcg/g (portio vaginalis). It was distributed in cervix uteri and portio vaginalis at highest concentration, followed by oviduct, tunica serosa and ovary, and to myometrium at lowest concentration. The same pattern of distribution was observed in transfer ratio of various location in uterus tissue to uterus arterial blood. The drug was transferred at very high level in pelvic cavity fluid after total hysterectomy throughout 3 postoperative hours. Five cases of gynecological infections receiving in total 10 to 42 g of CTX demonstrated 'excellent' results in 2 cases, and 'good' in 3 cases. Out of these, 3 cases did not respond to other antibiotic. Neither side effect nor clinical test abnormality was observed, except for one case, in which mild increase of GOT and GPT occurred. Based on the results of basic and clinical studies as described above, this drug is considered to have excellent efficacy and safety. PMID- 6270404 TI - [Clinical experience with cefotiam in the field of pediatrics (author's transl)]. AB - A clinical study on cefotiam (CTM) was performed in the field of pediatric infection, and the following results were obtained: The number of cases studied were six including 2 cases of U.T.I., 1 case of vaginitis and vulvitis, 1 cervical lymphadenitis, 1 absence of thigh and coxitis. Clinical responses to CTM were excellent in 3 cases, good in 1 cases but there was 1 exception who dropped out because of concurrent use of other antibiotics. As for bacteriological responses, CTM eliminated the pathogens E. coli and P. mirabilis from U.T.I., Staph. epidermidis and P. morganii, from vaginitis and vulvitis, H influenzae and Str. pneumoniae from cervical lymphadenitis and Staph. aureus from abscess of thigh. No side effect was observed except 1 case who showed an elevation of GOT, GPT and LDH. It is uncertain, however, wether this abnormality was resulted from the use of CTM or not, because the other antibiotics were used also. PMID- 6270405 TI - [Subacute toxicity study of cefotiam in three-week old beagle puppies (author's transl)]. AB - A subacute (5-week) subcutaneous toxicity study of cefotiam (CTM) was carried out using 9 three-week old Beagle puppies. The puppies were assigned to one of three groups, each containing three. Puppies in group I (control) were given physiological saline; puppies in group II and III were given 300 mg/kg/day of CTM and cefazolin (CEZ), respectively. No behavioral abnormalities were seen in puppies in each group. The changes, considered to be drug-related, were histopathological changes at the sites of injection, which consisted inflammatory cellular infiltration, hemorrhage and hyperplasia of fibroblast in connective tissue of skin and skeletal muscle. In terms of severity, CTM was rather more irritating than CEZ. Except the histopathological changes described above, there observed no abnormalities which were considered to be related to CTM. PMID- 6270406 TI - [Clinical studies of cefotiam in the field of pediatrics (author's transl)]. AB - The therapeutic effects of cefotiam (CTM) in the pediatric field were studied, and the following results were obtained: 1. Absorption and excretion: After 30 minutes intravenous drip infusion of 250 mg (14.7 mg/kg) of CTM to a 5-year old child weighing 17 kg, the serum levels and urinary excretion were measured. The serum levels were 47.8 mcg/ml at 30 minutes after administration, 23.9 mcg/ml after 1 hour, 12.5 mcg/ml after 1 hour and 30 minutes, 5.2 mcg/ml after 2 hours and 30 minutes, 0.3 mcg/ml after 6 hours and 30 minutes. Excretion rate in the urine was 48.8% at 6 hours after administration. 2. CLINICAL RESULTS: CTM was administered at a daily dose of 35 to 91 mg/kg b.i.d. to q.i.d. by intravenous or intravenous drip injection for 2 to 6.5 days to six patients i.e. two with bronchopneumonia, one with acute bronchitis, one with acute enteritis, one with impetigo and atopic dermatitis one with acute tonsillitis and acute cervical lymphadenitis. The responses were excellent in 2 cases and good in 4 cases. The response rate was 100%. No side effect was observed. PMID- 6270408 TI - [A clinical study of cefotiam in pediatric field (author's transl)]. AB - Cefotiam is a new semisynthetic cephalosporin antibiotic. The treatment with this drug was carried out in 12 cases of pediatric infection. The clinical response was observed in 92% of the cases. Cefotiam was found to be active against a variety of Gram-positive and Gram-negative bacteria. Neither marked side effects nor laboratory abnormalities were found with this treatment. PMID- 6270407 TI - [Studies on a new cephalosporin, cefotiam, in the pediatric field (author's transl)]. AB - The studies on cefotiam (CTM) administered by intravenous drip infusion in pediatric field made the following results: 1. The serum levels after administration of cefotiam 25 mg/kg, 63 mg/kg by intravenous was maintained high until 2 hours after and kept to be able to measure after 5 hours, and the half life was 0.9-1.7 hours. 2. The drug transferred actively about 1/5 of serum level into the intrathoracic pus. The excretion rate in urine was about 80% of the drug administered. 3. The clinical effective rate after the administration of CTM 50 70 mg/kg for 4 to 29 days was 95% in 21 patients with different five diseases. 4. No side effect and abnormality in laboratory findings were observed. PMID- 6270409 TI - [Clinical results of cefotiam in the field of pediatrics (author's transl)]. AB - We have administered cefotiam intravenously to 23 pediatric patients. The daily dose was 13-210 mg/kg. The clinical responses were excellent and good in 20 cases. Excluding 1 case with infectious mononucleosis, the efficacy rate of 90.9% (20/22 cases) was achieved As for side effect, diarrhea and eosinophilia were observed one each in 2 cases. In 2 cases, half lives were 24 and 53 minutes, urinary recovery rates were 88.3 and 91% over 6 hours. PMID- 6270410 TI - [Clinical experience with cefotiam (author's transl)]. AB - Cefotiam (CTM) were clinically applied to 6 cases and the following results were obtained: 1. The patients were aged 1 year in 5 cases and 7 years in 1 case. They comprised three cases of acute tonsillitis and 1 case each of phlegmon, bronchopneumonia and cervical abscess. CTM was administered in a daily dose of 50 115 mg/kg t.i.d. by an 1-hour intravenous drip infusion for 4 to 8 days. 2. The causative bacteria were S. aureus in 3 cases and H. parainfluenzae, K. pneumoniae and unknown pathogen in each 1 case. The organisms were eradicated in all cases except for only 1 case of S. aureus. 3. Overall responses were good in 4 cases and fair in 2 cases. 4. As for the abnormal laboratory findings, slight increase of GOT and GPT was observed in 1 case. Any other side effects were not observed. PMID- 6270411 TI - [Basic and clinical studies of cefotiam in the field of pediatrics (author's transl)]. AB - A new synthetic cephalosporin, cefotiam (CTM) was studied from the basic and clinical aspects, and the following results were obtained: 1. Bacteriological study: The bacterial activities of CTM against the clinical isolates of S. aureus, S. pyogenes, E. coli, Klebsiella sp. and Proteus sp. were compared with those of CEZ, CER, ABPC and GM. (1) As for S. aureus and S. pyogenes, the antibacterial actions of the conventional cephalosporins were slightly more potent than those of CTM. (2) However, CTM had the antibacterial actions which were most potent Proteus sp. among the above 4 antibiotics and more potent against E. coli and Klebsiella sp. than the above 2 conventional cephalosporins. 2. Pharmacokinetic study: The peak serum levels of CTM were comparatively low with 19.30 +/- 1.66 mcg/ml 30 minutes after a bolus injection of 20 mg/kg and 25.85 +/- 4.32 mcg/ml just after a drip infusion of 20 mg/kg. The half-life of the serum levels was 0.98 hrs. and 0.87 hrs., respectively. The 4 hours urinary excretions in six patients ranged from 38.6 to 64.9%, showing a slightly wide variance. 3. Clinical study: The clinical response was good in 23 out of total 25 cases, i.e. 14 cases with respiratory tract infection, 7 with urinary tract infection and 4 with skin and soft tissue infection. The response rate was 92%. Also, neither side effects nor abnormal laboratory findings was observed. PMID- 6270412 TI - [Clinical experience with cefotiam in pediatric infections (author's transl)]. AB - Cefotiam, one of the new cephem antibiotics, was used in 14 cases with pediatric infections: (10 cases with respiratory tract infections, 2 with urinary tract infections, each 1 with purulent meningitis + sepsis and acute appendicitis). The patients were aged between 15 days and 9 years old. The drug was, a rule, given at a daily dose of 50 mg/kg to 100 mg/kg q.i.d. by bolus intravenous injection. The duration of treatment was between 3 and 38 days. The treatment produced the following clinical responses: Out of the 10 cases, good response in 7 with respiratory tract infections, fair in 1 and poor in the remaining 2. The responses in urinary tract infections were excellent in 1 and good in the other case. An apparently clear response was obtained in 1 case with purulent meningitis + sepsis due to K. pneumoniae. Also, an excellent response was seen in 1 case with acute appendicitis. The response rate including fair response was 85.7%. The suspected pathogens isolated from 5 cases (S. aureus: 1. strain, H. influenzae: 1, K. pneumoniae 2, E. coli: 1) were eliminated after CTM administration. Good clinical responses were also obtained in these cases. No side effect was observed. Mild elevation of GOT and GPT was noted during the treatment in 1 case. It is unclear, however, if CTM was associated with this side effect or not. P. aeruginosa, Serratia appeared after superinfection in 1 case. PMID- 6270413 TI - [Clinical evaluation of cefotiam therapy in children (author's transl)]. AB - Cefotiam (CTM) was evaluated for its safety and efficacy in children. Twenty-six patients were treated with 40 to 200 mg/kg per day of CTM by intravenous administrations. The diagnosis of the patients were acute pharyngitis (2), acute bronchitis (1), pneumonia (4), empyema (2), urinary tract infection (2), typhoid fever (1), acute enterocolitis (2), partially-treated purulent meningitis (1), and suspected septicemia in neuroblastoma (1); and the remaining ten patients were considered to have nonbacterial infections. The pathogens recovered were Streptococcus pyogenes (1), Streptococcus pneumoniae (1), Staphylococcus aureus (4), Haemophilus influenzae (4), Escherichia coli (1), enteropathogenic Escherichia coli (1), Salmonella typhi (1), and Campylobacter jejuni (1). All but two patients of bacterial infections were cured after the CTM therapy, and the rate of efficacy was 87.5%. Diarrhea (3), urticaria (1), transient elevation of GOT and GPT (1), and transient eosinophilia (3) were found to be associated with the CTM therapy. However, no severe adverse reactions were encountered. Half life of the serum CTM level was 0.93 +/- 0.13 hours, and excretion into the urine was rapid. CSF concentration obtained 1 hour after an intravenous injection of 21 mg/kg of CTM in a case with inflamed meninges was 1.5 mcg/ml, and the CSF/serum ratio was 9.0%. From these data, CTM appears to be a safe and effective antibiotic when used in children with susceptible bacterial infections. PMID- 6270414 TI - [Influence of cefotiam (SCE-963, CTM) on bowel flora (author's transl)]. AB - The influence on bowel flora of CTM was studied in 5 children who were taking normal diet. 1) In the cases following no diarrhea, administration of CTM caused no significant changes in bowel flora. In the cases following diarrhea, administration of CTM caused a fall in coliform, BEP group, Lactobacillus and Peptostreptococcus. However, after the administration was discontinued, the reduced bowel flora was returned to the normal range within a few days. 2) No overgrowth of bowel flora by Pseudomonas, Clostridium difficile or Candida was observed. 3) One strain of nontoxigenic Clostridium difficile was observed in the case of following no diarrhea. 4) As for the side effects, diarrhea was observed in 2 cases. BEP group: Nonspore making anaerobic Gram positive rods of Bifidobacterium, Eubacterium, Propionibacterium. PMID- 6270415 TI - [Fundamental and clinical studies of cefotiam in the field of pediatrics (author's transl)]. AB - The fundamental and clinical studies on cefotiam (CTM) were performed in the field of pediatrics, and the following results were obtained: 1. The peak MIC's of CTM against Gram negative rods such as E. coli, Klebsiella, Salmonella, Proteus were 1.56 mcg/ml. The MIC distribution against S. aureus was almost equal to the conventional cephalosporin antibiotics. The MICs against P. pseudomonas and Serratia were over 400 mcg/ml. 2. The mean serum levels of CTM after bolus intravenous injection of 25 mg/kg were 59.9 mcg/ml after 15 min., 30.0 mcg/ml after 30 min., 15.6 mcg/ml after 1 hour. 3. Administration of CTM to 6 pediatric patients produced the clinical responses which were good in all 6 cases and the bacterial effects of eradication in 3 cases and superinfection in the 2 cases in the 5 cases from whom the organism were isolated. No side effect was observed. From the above results, it is considered that a bolus injection of CTM 25 mg/kg t.i.d. to q.i.d. is a safe and useful treatment for pediatric cases. PMID- 6270416 TI - [Cefotiam preparations]. PMID- 6270417 TI - [Evaluation of cefotiam in pediatric field (author's transl)]. AB - Studies on the antibacterial activity, absorption and excretion and also clinical investigation in the field of pediatrics have been carried out with cefotiam (SCE 963, CTM), a new cephalosporin antibiotic. 1) The MICs of CTM against the following clinical isolates were measured and compared with those of CEZ: S. aureus (81 strains), E. coli (27) and K. pneumoniae (27), with CTM being inferior by 1 tube in S. aureus, being superior by 2 to 3 tubes in E. coli and by about 2 3 tubes in K. pneumoniae. 2) Absorption and excretion. After intravenous one shot injection at dose levels of 10 mg/kg and 20 mg/kg, the peak in the serum concentration was shown in the 15-minute value with 18.1 and 36.6 mcg/ml for 10 mg/kg and 20 mg/kg, respectively. The half-life in ;the serum was 1.14 and 0.61 hours, respectively. In the case of 1-hour intravenous drip infusion at a dose level of 10 mg/kg, it was 14.3 mcg/ml, with 0.98-hour half-life in the serum. The recovery rates from the urine within 0 to 6 hours were 50.6% and 66.2% in the case of intravenous one shot injection at dose levels of 10 mg/kg and 20 mg/kg, respectively, with 71.1% in the case of the 1-hour intravenous drip infusion. 3) Two to 3 hours after intravenous one shot injection of CTM in H. influenzae meningitis every 4 hours at a dose level of 62.5 mg/kg at one time, the cerebrospinal fluid concentration of CTM was only 2.12 to 10.0 mcg/ml, and this fact suggests that CTM is a useful cephalosporin for treating purulent meningitis. 4) CTM was administered in 19 clinical cases, with the results being: excellent in 4 out of 4 cases of bronchitis; excellent in 5 and good in 1 out of 6 cases of pneumonia; excellent in 3 cases of pyelitis; good in purulent parotitis, purulent meningitis and bacterial pericarditis; and excellent in peritonsillar abscess, purulent osteomyelitis and staphylococcal scalded skin syndrome (S.S.S.S.). No side effects have been observed in all cases. As for abnormal laboratory findings, 3 cases of eosinophilia were seen. PMID- 6270418 TI - [Clinical studies on cefotiam in pediatric infections (author's transl)]. AB - Our investigation of cefotiam in pediatric infection produced the following results: 1. Cefotiam was administered intravenously by one shot or drip infusion in 20 patients with infectious diseases. These diseases consisted of 13 pneumonia, 3 upper respiratory tract infections, 3 pyelonephritis, 2 other urinary tract infections and one purulent meningitis. Cefotiam was effective in all cases. 2. Transient elevation in serum GOT, GPT, Al-P and LDH was observed in 3 cases. But other side effect was not noted in any cases. PMID- 6270419 TI - [Basic and clinical studies of cefotiam in pediatric field (author's transl)]. AB - The basic and clinical studies of cefotiam (CTM) in pediatric infections were carried out, and the following results were obtained: 1. The antibacterial activity of CTM against S. aureus was equal or slightly less than that of cefazolin (CEZ). Those of CTM against E. coli and K. pneumoniae were eight times more active than those of CEZ. 2. CTM 20 mg/kg was administered wither by 30 minutes or 1 hour intravenous drip infusion. The peak serum levels were obtained at the end of each drip infusion, with the mean peak levels being 44.8 and 41.4 mcg/ml respectively. The serum levels at 1.5 and 2 hours after drip infusion were 2.8 and 2.2 mcg/ml respectively, and at 3.5 and 4 hours after drip and 4 hours after drip infusion were 0.3 and 0.7 mcg/ml respectively. The half lives were 0.62 and 1.15 hours, respectively. The mean urinary excretion over 6 hours were 52.8% in ;the 30 minutes drip infusion group and 42.6% in the 1 hour drip infusion group. 3. Clinical efficacy was evaluated in sixteen cases suffering from tonsillitis (4 cases), pneumonia (4), bronchitis (2), cervical lymphadenitis (2), purulent meningitis (2), suppurative arthritis (1) and suspected sepsis (1). Good and excellent responses were obtained in 15 of 16 cases (93.8%). Bacteriological response in the form of eradication was noted in 4 of 6 cases. Side effect observed was rash in 1 case, and laboratory abnormalities were elevation of BUN in 1 case and elevation of GPT in 2 cases. PMID- 6270420 TI - Clinical evaluation of cefotaxime in the treatment of purulent meningitis in children. PMID- 6270421 TI - [Clinical evaluation of cefotiam in children (author's transl)]. AB - Clinical trial of cefotiam was made in children and the following results were obtained. 1. Pharmacodynamic studies of the drug in CSF of experimental staphylococcal meningitis in rabbits showed a CSF/serum ratio of T 1/2 of 3.52, which was relatively high, but a percentage of CSF/serum ratio of AUC of only 3.42% up to 3 hours, suggesting a low efficiency of passage of the drug into CSF. 2. Blood concentrations of the drug were determined in children after an intravenous bolus injection of 20 mg/kg and were 46 mcg/ml (15 min.) and 26 mcg/ml (30 min.), T 1/2 being 40.8 min., Urinary recovery rate was 91.3% up to 4 hours in one patient and 61.9% up to 6 hours in another, respectively. 3. Thirteen patients with the following 14 episodes of infections were treated with cefotiam; urinary tract infection (5 cases), pneumonia (5), empyema (1), tonsillitis (1) and suspected sepsis (2). An overall efficacy rate was 100%, i. e., excellent in 12, good in 2 and no failure. No adverse reactions were clinically discernible and only laboratory abnormalities were transient or slight elevations of transaminase levels in 2 patients. 4. Based on the above results, it was concluded that cefotiam is a potent new antibiotic in the treatment of bacterial infections. Spectrum and antibacterial activity of the drug suggest that the drug particularly indicated for pneumonia. PMID- 6270422 TI - [Changes in food habits among the Japanese and neoplasms in the large intestine]. PMID- 6270423 TI - [Histological pattern and prognosis in colorectal neoplasms]. PMID- 6270424 TI - [Histopathological diagnosis of biopsy of endometrium (author's transl)]. PMID- 6270425 TI - [Angiographic evaluation of hepatocellular hepatoma for hepatic resection (author's transl)]. PMID- 6270426 TI - [Lung tumor: gallium 87 citrate and thallium 201 chloride (author's transl)]. PMID- 6270427 TI - [Clinical evaluation of 67Ga-citrate scintigraphy for pulmonary carcinomas (author's transl)]. PMID- 6270428 TI - [Late complication of radiation treatment of malignant lymphomas; especially on a consideration of adjoining therapy fields (author's transl)]. PMID- 6270429 TI - [Necrolytic migratory erythema in glucagonoma syndrome (author's transl)]. PMID- 6270430 TI - [RI angiography with in vivo labeled 99mTc-pertechnetate RBC in the specific diagnosis of cavernous hemangioma of the liver (author's transl)]. PMID- 6270431 TI - [Potassium transport in the rat intestine. The role of Na-K-ATPase (author's transl)]. PMID- 6270432 TI - [Studies on in vivo labeling rate of red blood cells eith 99mTc-pertechnetate in radionuclide angiography (author's transl)]. PMID- 6270433 TI - Electrophysiological studies on plasticity of cerebellothalamic neurons in rats following neonatal hemicerebellectomy. AB - Changes in the cerebello-thalamo-cerebral projection following hemicerebellectomy were investigated in 16 rats by laminar field potential analysis of cerebello cerebral responses in the cerebral cortex and by unitary recording of antidromic responses of cerebellar nuclear neurons to stimulation of the thalamus. In contrast to the occurrence of an exclusively contralateral cerebello-cerebral response in intact rats, a significant bilateral cerebello-cerebral response could be evoked in all 8 rats hemicerebellectomized before 6 days of age (early hemicerebellectomy). On the other hand, in all 5 rats operated on at 8 to 15 days of age (late hemicerebellectomy) and in the 3 rats operated on in adulthood, the cerebello-cerebral response was evoked only contralaterally just as in intact rats. Unitary recordings of antidromic responses of cerebellar nuclear neurons revealed that bilateral or ipsilateral cerebellothalamic projection neurons were remarkably more numerous in early hemicerebellectomized rats than in controls (P less than 0.001). Such neurons were less frequently found in late hemicerebellectomized rats, but they were still more numerous than in controls (P less than 0.005). Absence of the ipsilateral cerebello-cerebral response in late hemicerebellectomized rats indicates that even though bilateral or ipsilateral cerebellothalamic projection neurons may exist in these rats more numerously than in intact rats, they may not be sufficient in number or in efficiency to mediate a detectably response. PMID- 6270434 TI - Effect of procaine amide on the membrane currents of the sino-atrial node cells of rabbits. AB - Using small rabbit sino-atrial node preparations, the effects of procaine amide in concentrations from 0.01 to 2 mg/ml on the membrane potentials currents were studied by both current-clamp and voltage-clamp experiments. Procaine amide in concentrations over 0.1 mg/ml reduced the peak of the action potential, maximum diastolic potential and the maximum rate of depolarization. The action potential duration was prolonged, the resting potential was decreased and the heart rate was reduced. In the voltage-clamp experiments, procaine amide (0.1 mg/ml) reduced the slow inward current (is), the outward current (iK) and the inward current activated by hyperpolarization (ih). The major effect, however, was the reduction of the outward current. Sine the degree of the steady-state activation of iK and its time constant were unchanged, the observed reduction of iK could have been caused by a reduction of iK. PMID- 6270435 TI - [The role of the functional maturation of alveolar macrophages and alveolar lining materials in lung defenses (author's transl)]. PMID- 6270436 TI - [Eighteen years experience of lung cancer resection at the National Cancer Center Hospital, Tokyo (author's transl)]. PMID- 6270437 TI - [Lung cancer: problems in the treatment of advanced lung cancer (author's transl)]. PMID- 6270438 TI - [A case of hypersensitivity pneumonitis presumably due to Thermoactinomyces vulgaris with associated uveitis and serum-angiotensinconverting enzyme elevation (author's transl)]. PMID- 6270439 TI - [Mucinous adenocarcinoma of the renal pelvis with other two cancers in urologic organs, triple cancer (author's transl)]. PMID- 6270441 TI - Growth characteristics of feline panleukopenia virus in synchronized kitten kidney cells. PMID- 6270440 TI - Synergistic effect of Sendai virus on Mycoplasma pulmonis infection in mice. PMID- 6270442 TI - Evidence on horizontal transmission of bovine leukemia virus due to blood-sucking tabanid flies. PMID- 6270443 TI - [99mTc-pyrophosphate scintigraphic indices at different times in acute myocardial infarct]. AB - The incorporation rate of 99mTc-pyrophosphate into the myocardium was studied in 123 patients with acute myocardial infarction at different stages of the disease. The capacity of the damaged myocardium to accumulate the labelled pyrophosphate decreased with its healing. The accumulation character of the radiopharmacologic preparation also changed. The revealed phenomena are explained by the features of the 99mTc-pyrophosphate concentration mechanisms of the damaged areas of the myocardium. during the necrosis phase pyrophosphate is accumulated in the irreversibly damaged myocardial cells. The incorporation of 99mTc-pyrophosphate into the myocardium at the late stages of the disease are explained by both the presence of the necrotized cells in the myocardium and possible binding of the radiopharmacologic preparation to collagen. PMID- 6270444 TI - [Dynamics of 5'-nucleotidase activity in the blood plasma in experimental myocardial infarct]. AB - The authors studied in the bloodflow the appearance of the damaged external cellular membranes of the heart during the development of experimental myocardial infarction. The appearance of the external cellular membranes was judged by the activity of their specific marked 51-nucleotidase. It was established that myocardial infarction leads to increased activity of 51-nucleotidase in the blood plasma, which is retained for about 1 month. The maximum of activity of 51 nucleotidase reflects the most intensive resorption of the damaged cardiac tissue, and is seen from 3 to 7 days after the accident. During these time limits thrombotic complications are especially dangerous. PMID- 6270445 TI - Angiotensin receptors in glomeruli differ from those in renal arterioles. AB - Angiotensin receptors in afferent and efferent arterioles and in the glomerulus are strategically located to influence renal perfusion and glomerular function. With the size of isolated glomeruli as the index, we have demonstrated identical dose-response relationships for graded concentrations (10(-13) to 10(-3) g/liter) of angiotensin II (AII) and angiotensin III (AIII). An octapeptide analogue (saralasin 10(-6) to 10(-2) g/liter) was equally effective at blocking glomerular responses to both AII and AIII, but two heptapeptide analogues (des-asp, 8-ile AII and des-asp, 8-gly AII; 10(-6) to 10(-2) g/liter) failed to block responses to either agonist. The relative influence of octapeptide and heptapeptide analogues on GFR was examined in anesthetized dogs with partial occlusion of the thoracic inferior vena cava. In 18 dogs, caval occlusion reduced renal blood flow (35%), GFR (29%), and arterial pressure (13%). Saralasin (300 to 3000 ng/kg/min, i.v.) and des-asp, 8-ile AII (100 to 3000 ng/kg/min, i.v.) increased renal blood flow by 0.41 +/- 0.11 and 0.62 +/- 0.11 ml/g/min, respectively, but only the octapeptide induced a concomitant increase in GFR (octapeptide: delta GFR = 0.11 +/- 0.03 ml/g/min; heptapeptide: delta GFR = -0.08 +/- 0.07 ml/g/min; P less than 0.025). As octapeptide and heptapeptide analogues were equally effective on renal blood flow in this and in previous studies, but only the octapeptide was effective in isolated glomeruli and in increasing GFR in the intact animal, we conclude that renal vascular and glomerular receptors differ. Furthermore, the glomerular receptor may be the more important in modulating the glomerular functional response to angiotensin. PMID- 6270446 TI - Phosphatase activity along the nephron of mice with hypophosphatemic vitamin-D resistant rickets. PMID- 6270447 TI - Responsiveness of plasma renin and aldosterone in diabetes mellitus. PMID- 6270448 TI - [Cell membrane characteristics and biological behavior of virus transformed cells (author's transl)]. AB - The transformation of a normal into a tumor cell is not caused by a single molecular event, but is the consequence of several simultaneous or consecutive molecular processes, which lead to a variety of changes in the structure and the metabolism of the cell. Investigations with the Rous sarcoma virus show that a single gene is primarily responsible for these changes that is coding for a phosphoprotein which, however, is multifunctional. The biochemical and biologic events which initiate and maintain the transformed status of the cell involve mainly the cytoplasma membrane. At both the outer and the inner surface of the cell membrane dramatic changes occur which influence the cell structure, permeability of the cytoplasma membrane, and the intracellular metabolic pathways. Most probably, these transformation-associated events are also involved in cell proliferation under physiologic conditions. In the tumor cell, however, they are not further regulated physiologically, with the consequence of an uncontrolled and incessant cell division. PMID- 6270450 TI - [Treatment and prognosis of superior vena caval syndrome in small-cell anaplastic carcinoma of the lung (author's transl)]. AB - One hundred and forty nine consecutive patients with small-cell anaplastic carcinoma of the lung were studied retrospectively. 19 patients (13%) presented with superior vena caval obstruction, 11 further patients (7%) developed this complication during the course of their disease. 16 patients presenting with superior vena caval syndrome were treated with chemotherapy alone and 13 patients responded. Due to suboptimal chemotherapy for current standards 7 of the 13 successfully treated patients (54%) relapsed after a median of 4.5 months. 3 patients were effectively retreated with another chemotherapy combination, 4 patients with radiation therapy. Superior vena caval obstruction is a frequent complication of small-cell anaplastic carcinoma of the lung. It can be treated effectively with combination chemotherapy. PMID- 6270451 TI - [Blood thyrotropin, thyroxine and adenosine cyclic monophosphate in peptic ulcer]. PMID- 6270449 TI - Receptor dysfunctions in human disease. AB - The characterization of binding parameters of hormones and drugs to specific receptors at various human cell types have introduced an interesting approach for the evaluation of pathogenic mechanisms involved in endocrine and metabolic disorders. The dysregulation of cellular receptors in those disease include changes in receptor number, changes in binding affinity and production of antibodies against receptor molecules. It can be concluded from these observations that altered receptor physiology may be of important value for abnormalities in cellular recognition and control mechanisms which can be observed in neoplastic, inflammatory, immune and developmental diseases. PMID- 6270452 TI - [Role of cyclic nucleotides in the regulation of immunity in Salmonella infections]. PMID- 6270454 TI - ["Rearing" of prematurely born and full-term infants]. PMID- 6270453 TI - [Effect of radiations with varying linear energy transfer on the skin of mice]. AB - Experiments were carried out to measure the time and severity of the radiation reaction of the skin of mice exposed to X- and gamma-radiations, protons with energies of 645 and 50 MeV as well as accelerated helium ions at doses of 200 to 4000 rad. It was found that relative biological effectiveness coefficients of 645 and 50 MeV protons were 1.0 and those of helium ions were 1.3 for the skin reaction at early and late stages of observation. No significant difference in the time of manifestation of radiation-induced skin lesions as related to radiations with various LET was detected. PMID- 6270455 TI - [An example of rehabilitation thanks to team work]. PMID- 6270456 TI - [Operating room nurses in Switzerland]. PMID- 6270457 TI - [Health 80. Survey of households in Geneva]. PMID- 6270459 TI - [Therapeutic strategy]. PMID- 6270458 TI - [Articular chondrocalcinosis]. PMID- 6270460 TI - [Nursing care of a dying patient with chronic respiratory insufficiency]. PMID- 6270461 TI - [Neurotic depression in children of "internally rejecting" mothers]. PMID- 6270462 TI - [The meaning of the diagnosis cancer]. PMID- 6270463 TI - [What does "mental health" mean?]. PMID- 6270464 TI - [Informating the laryngectomized patient and his family]. PMID- 6270465 TI - [Handicapped love]. PMID- 6270466 TI - [Death of my mother]. PMID- 6270467 TI - Lethal pneumonia in guinea pigs associated with a virus. AB - No bacteria were observed in an epizootic of lethal pneumonia in guinea pigs. Necrotic bronchitis and bronchiolitis with basophilic intranuclear inclusion bodies in bronchial epithelial cells were characteristic. Although adenovirus infection of guinea pigs has not previously been reported, histological findings paralleled those found in adenovirus infections of other animals including man. Virus particles found by electron-microscopical examination of the lung tissue closely resembled adenoviruses. The disease seemed to have a low contagiousness, a low morbidity (about 0.7%), but an acute course and a high mortality (100%). PMID- 6270468 TI - Hormone receptors in male breast carcinoma. AB - It is well established that approximately 60% of female breast cancers contain estrogen receptors (ER+). The hormonal receptor status of male breast cancers remains relatively unknown. Tumor tissue from 14 patients has been reviewed at the University of Oregon Health Sciences Center. Specimens of all 14 patients were ER+ with a range of 5.5 to 374 fmol/mg protein. Eight of 12 patients had tumors which also contained progesterone receptors (PR+) with a range of 42 to 1852 fmol/mg protein. Nine patients were treated by modified radical mastectomy, two by radical mastectomy, one by simple mastectomy, and two by biopsies only. Twelve patients remain free of disease with a mean follow-up of less than 2 years. The two patients who developed metastases both responded to endocrine ablation by orchiectomy and subsequent adrenalectomy. These data suggest that male breast cancer is highly endocrine sensitive and that endocrine ablation can play an integral part in those men unfortunate enough to develop disseminated disease. PMID- 6270469 TI - Studies on the role of oral contraceptive use in the etiology of benign and malignant liver tumors. PMID- 6270470 TI - Heme utilization by rat liver tryptophan pyrrolase as a screening test for exacerbation of hepatic porphyrias by drugs. AB - Rat liver tryptophan pyrrolase plays a versatile and unique role among hepatic hemoproteins in relation to heme utilization. The depletion of pyrrolase heme in the experimental porphyria produced by 3,5-diethoxycarbonyl-1,4-dihydrocollidine is potentiated by joint administration of any one of 19 drugs known to exacerbate the human disease, but not by any of 13 nonexacerbators. These findings form the basis of a screening test for drug exacerbation of hepatic porphyrias; the conditions, details, and requirements of which are described. PMID- 6270471 TI - Patterns of cyclic nucleotides and related enzymes in normal and Gross-virus transformed rat thymocytes. PMID- 6270472 TI - A case of lymphedema of the lower limbs as a result of metastatic spread to the lymphatic vessels of the dermis. PMID- 6270473 TI - Indirect caudal lymphography using a new water-soluble contrast agent--animal experimental studies in pigs. PMID- 6270474 TI - Identifying hospital patients who need early discharge planning for special dispositions: a comparison of alternative techniques. PMID- 6270475 TI - Respiratory infection and airway reactivity. AB - The data reviewed demonstrate that viral and mycoplasma infections induce a spectrum of functional abnormalities in airways. Acute virus infections cause wheezing illnesses in both children and adults. Changes in peripheral airway function during infection with similar organisms are observed in other subjects, usually normal adults. The pathogenesis of these responses is unclear. Pathologic data do show that infection with these pathogens damages the airway epithelium. These changes appear to increase permeability of the respiratory epithelium to protein antigens and consequently may contribute to increased frequency of attacks in asthmatic subjects. In addition, increased mucosal permeability may enhance delivery of inhaled drugs to effector sites in airway walls to induce exaggerated bronchoconstrictor responses in clinical challenge situations. Whether changes in the epithelium during infection, inducing greater antigen entry into the interstitium, results in subsequent development of specific allergy is not known and requires further study. PMID- 6270476 TI - [Zonal psoriasis]. AB - Two cases of Zonal Psoriasis are reported. The lesions cleared after topical chrysarobin. The onset and the course of this unusual form point out a neurogenic relationship. PMID- 6270477 TI - Treatment of vitreous opacities in a case of familial amyloidotic polyneuropathy by vitreous surgery. PMID- 6270478 TI - The sublabial approach for extensive nasal and sinus resection. PMID- 6270479 TI - [Angiotensin converting enzyme (ACE) in the serum of some respiratory patients (author's transl)]. PMID- 6270481 TI - Characteristics of [3H]1 alpha, 25-(OH)2D3 binding to nuclear fractions from rat pituitary adenoma GH3 cells. PMID- 6270480 TI - Comparison of the anticonvulsant effects of opioid peptides and etorphine in rats after icv administration. PMID- 6270482 TI - Inhibitory and stimulatory action of vasopressin on the secretion of corticotrophin in rats : structure-activity study. PMID- 6270483 TI - Asbestos-elicited catecholamine secretion from isolated bovine adrenal chromaffin cells. PMID- 6270484 TI - Effect of beta-endorphin on the steroid production of isolated zona glomerulosa and zona fasciculata cells. PMID- 6270485 TI - Influence of cyclic GMP on rodent aggressive behavior. PMID- 6270487 TI - Effect of chronic alcohol administration on the hormonal sensitivity of isolated perfused rat liver. PMID- 6270486 TI - Improved high-affinity binding of 3H-apomorphine with a subcellular membrane fraction of mammalian brain tissue. PMID- 6270488 TI - Soluble gamma-aminobutyric acid and benzodiazepine receptors from rat cerebral cortex. PMID- 6270489 TI - The sensitivity of opiate receptors and ligands to short wavelength ultraviolet light. PMID- 6270490 TI - A radioreceptor assay for opiate drugs in human cerebrospinal fluid and plasma. PMID- 6270491 TI - Multiple benzodiazepine receptors and their regulation by gamma-aminobutyric acid. PMID- 6270492 TI - Withdrawal symptoms in morphine-dependent rats intracerebroventricularly injected with ACTH1-24 and with beta-MSH. PMID- 6270493 TI - Phospholipid synthesis in human embryo fibroblasts infected with herpes simplex virus type 2. AB - The effect of herpes simplex virus type 2 infection on the synthesis of phospholipids in human embryo fibroblasts was determined at temperatures permissive (35 C) or nonpermissive (42 C) for virus replication. Incorporation of [32P]i was decreased by herpes simplex virus type 2 infection after 6 hr, which corresponds to the time of initiation of progeny virus production. No differences were observed in the relative incorporation of [32P]i phospholipid classes. In another series of experiments cells were labeled with [3H]ethanolamine before infection and with [14C]ethanolamine after infection. The incorporation of [14C]ethanolamine was also decreased after 6 hr of infection. When choline was substituted for ethanolamine, a similar, although less pronounced, decrease in incorporation was seen in infected cells compared to mock-infected cells. During abortive infections at 42 C, incorporation of [3H]thymidine into cellular DNA was stimulated, but the incorporation of phospholipid precursors was decreased. Total phospholipid composition and phospholipid acyl group composition were not changed appreciably during abortive or productive infection, regardless of whether the cells were labeled before or after infection. In conclusion, these data indicated that, during herpes simplex virus type 2 infection, the incorporation of lipid precursors into phospholipid was decreased. The stimulation of cellular DNA synthesis previously observed during abortive infection at 42 C was not paralleled by a detectable stimulation of total phospholipid synthesis. Neither productive nor abortive infection resulted in significant phospholipid compositional changes in the host cell; however, both resulted in a marked inhibition of phospholipid synthesis. PMID- 6270495 TI - [Morphological assessment of the effectiveness of variants in preoperative gamma therapy of rectal cancer]. PMID- 6270494 TI - Effect of Ca++ on triphosphoinositide extraction in fusing myoblasts. AB - Ca++-dependent degradation of triphosphoinositide has been postulated to regulate levels of membrane-bound Ca++ and to generate a 1,2-diacylglycerol fusogen in cell fusion. Triphosphoinositide metabolism was therefore studied during Ca++ induced fusion of cultured chick embryo myoblasts. Using a frequently cited extraction procedure, it was found that apparent Ca++-dependent triphosphoinositide degradation was actually due to inhibition of extraction. A new procedure using the ion-pairing reagent tetrabutylammonium sulfate was developed which was unaffected by Ca++ and gave 2- to 20-fold greater extraction of triphosphoinositide than existing procedures. With this procedure, no changes in triphosphoinositide metabolism were found during myoblast fusion. PMID- 6270496 TI - A mathematical pattern of the evaluation of biological aggressiveness of silica dusts. PMID- 6270497 TI - The ultrastructure of osteoclasts in microphthalmic mice. AB - Osteoclasts in microphthalmic (mi/mi) mice were significantly smaller than in their heterozygous (mi/+) or normal (+/+) littermates as measured by electronmicroscopic morphometry. Clear zones were also significantly smaller and normal ruffled borders were absent. The cytoplasm was not fully differentiated. No differences were seen between osteoclasts from mi/+ and +/+ mice. The number of osteoclasts was not different in mi/mi, mi/+ or +/+ mice. Lysosomal enzymes and collagenase content of bones in the three genotypes did not show marked differences. PMID- 6270498 TI - The pathogenesis of infantile malignant osteopetrosis: bone mineral metabolism and complications in five infants. AB - Bone mineral metabolism was studied in five infants aged 8 to 22 months with severe osteopetrosis. There were findings consistent with biochemical osteomalacia. These included hypocalcemia, hypophosphatemia, high serum acid phosphatase and alkaline phosphatase activity, high levels of serum parathyroid hormone, and high urinary cyclic AMP. Serum 1,25(OH)2 vitamin D3 level was high in the one patient tested. Radiographs in all infants revealed rachitic changes in the metaphyses. However, dense bones on radiographs, calcium balance studies, and radio-calcium absorption studies demonstrated markedly positive calcium balance. Iliac crest bone biopsies showed increased quantity of woven bone with abundant numbers of osteoclasts, excessive amounts of osteoid, myelofibrosis, and a decreased number of Howship's lacunae. The wide bands of unmineralized osteoid did not take up tetracycline. In vitro bone resorbing activity due to osteoclast activating factor from cultured stimulated leukocytes was normal. Bone turnover however, was now as evidenced by low urinary hydroxyproline levels. We interpret these findings as indicating there is decreased bone remodeling and resorption in spite of increased humoral stimuli and osteoclasts. Since calcitonin levels were normal for age, the most likely cause of the impaired bone remodeling sequence was defective osteoclast function. We postulate that there may be a common genetic defect in phagocyte cells, including monocytes, neutrophils and osteoclasts, which accounts for the abnormalities of mineral metabolism and previously reported hematologic, neurologic, and infectious complications. PMID- 6270499 TI - The effect of fasting on liver receptors for prolactin and growth hormone. AB - The effects of 3 day fasting on liver prolactin and growth hormone receptors have been investigated in male and female rats. Fasting caused a significant fall in serum immunoreactive insulin (67% decrease), while receptor-reactive somatomedin fell 82% when measured in whole serum and by 72% when measured in serum fractions following gel chromatography at low pH. Tracer ovine prolactin binding to liver microsomal membranes was reduced by 55% on fasting in females, but unchanged in males. Tracer bovine growth hormone binding fell significantly in both sexes. Analysis of competitive binding curves showed the decrease binding to be due to a loss of prolactin receptors in females, and of high affinity (but not low affinity) growth hormone receptors in males and females. Significant correlations were seen between serum insulin and tracer prolactin (females) and growth hormone (males and females) binding to liver membranes. Correlations between serum insulin and liver high affinity growth hormone binding sites were particularly significant (r = 0.899 in females, r = 0.910 in males). It is proposed that the hypoinsulinemia of fasting causes a loss of high affinity growth hormone receptors in the liver, which could result in a relative hepatic resistance to growth hormone and a decreased hepatic generation of somatomedin. PMID- 6270500 TI - Meal stimulation of cortisol secretion: a protein induced effect. AB - Cortisol and ACTH secretion was studied in 52 healthy subjects who were fasted of fed various diets: standard, high fat, high carbohydrate, high protein. Subjects fed high protein diet (4 gm/kg body weight) showed significant increases in cortisol both at 30 and 60 min after the 1200 hr meal and 30 min after the 1600 hr meal. Increases in cortisol, of a smaller magnitude, were also seen after both the 1200 and 1600 hr meals in each of the diets with 1 gm protein/kg body weight (standard, high fat, high carbohydrate). ACTH was significantly increased following the 1200 hr and 1600 hr meals with the high protein diet. We conclude that dietary protein plays an important role in meal stimulated cortisol release. PMID- 6270501 TI - An IS4-encoded protein is synthesized in minicells. AB - A protein of Mr 47,000 is synthesized in Escherichia coli minicells, when these harbor a multicopy plasmid carrying IS4 in either orientation and between different flanking sequences. The protein corresponds to the sequence predicted from the known DNA sequence of IS4, as shown by partial N-terminal radiolabel protein sequence analysis. Its apparent molecular weight, however, as determined from its electrophoretic mobility in SDS polyacrylamide gels, is smaller than predicted. When compared with other plasmid-encoded proteins, the IS4-encoded protein is synthesized in minicells in small amounts. Its synthesis has not been detected in a DNA-dependent cell-free system. PMID- 6270502 TI - DNA sequence homology in Rhizobium meliloti plasmids. AB - Plasmids were recovered by an alkaline procedure from six symbiotically effective strains of Rhizobium meliloti of diverse geographical origin, reported to harbour only one middle-size large plasmid (ranging from 89 to 143 Megadaltons). Each purified plasmid was digested with eight restriction endonucleases; cleavage patterns were very complex: only KpnI and XbaI gave a limited number of bands. Fingerprints were very different, whatever the restriction enzyme or the geographical origin of the strains. However, Southern DNA-DNA hybridizations revealed that the plasmids showed homologous sequences having a high thermal stability. We gave evidence that some of these sequences are common to all the plasmids of R. meliloti. The biological function of these common sequences is unknown. Hybridization with cloned nitrogen fixation (nif) genes from Klebsiella pneumoniae had demonstrated that nif genes were not located on the middle -- size plasmids of R. meliloti studied in this paper. PMID- 6270503 TI - Isolation of an Hfr donor of Pseudomonas aeruginosa PAO by insertion of the plasmid RP1 into the tryptophan synthase gene. AB - A derivative of the IncP-1 plasmid RP1, temperature-sensitive for maintenance, was inserted into the Pseudomonas aeruginosa chromosome by selection for a plasmid marker (carbenicillin resistance) at non-permissive temperature. In one strain, PAO 1000, the plasmid was stably integrated in the trpA, B gene cluster mapped at 27 min, as shown by the following evidence. (i) Trp+ transductants lost all plasmid markers. (ii) Cleared lysates of PAO 1000 showed no plasmid band typical of the autonomous RP1 in agarose gel electrophoresis. (iii) No transfer of carbenicillin resistance by PAO 1000 was detectable. (iv) PAO 1000 mobilised the chromosome from an origin at, or very near, the plasmid insertion site with high frequency (recovery of proximal markers greater than or equal to 10(-3) per donor). Matings on the plate with and without interruption of conjugation showed that chromosome transfer was unidirectional. (v) Recombinants from PAO 1000 mediated crosses did not inherit plasmid markers or the trpA, B mutation. A derivative of PAO 1000 was obtained which had lost the Hfr property and all plasmid markers except carbenicillin resistance. This strain (PAO 1001), when carrying the autonomous RP1 plasmid, was capable of unidirectional chromosome mobilisation like PAO 1000, but with 50-fold lower efficiency. We propose that integration of the temperature-sensitive RP1 plasmid in PAO 1000 occurred via transposition of Tn1, the element specifying carbenicillin resistance. PMID- 6270505 TI - The cloning of Aspergillus nidulans mitochondrial DNA in Escherichia coli on plasmid pBR322. PMID- 6270504 TI - Restriction endonuclease cleavage map of pKM101: relationship to parental plasmid R46. AB - A detailed restriction endonuclease cleavage map of the plasmid pKM101 has been constructed. pKM101 plasmids containing individual Tn5 insertions were used to facilitate the ordering of restriction fragments generated by enzymes cleaving pKM101 at multiple sites. By restriction enzyme analysis, pKM101 (35.4 kilobases) appears to have arisen from its clinically-isolated parent by deletion of a single DNA region which codes for three of the four drug resistances carried by R46. PMID- 6270506 TI - The replication of Escherichia coli chromosome can be initiated by integrated ColE1-type plasmids. PMID- 6270508 TI - Isolation of an Hswl Nav4 influenza virus from a tufted duck (Aythya fuligula) in Japan. AB - A hemagglutinating agent was isolated from a tufted duck captured in Lake Shinji, Shimane Prefecture, Japan, and identified as influenza A by double immunodiffusion tests with the antiserum to influenza A virus ribonucleoprotein. The hemagglutinin in the isolate was antigenically related to that of A/New Jersey/8/76 but was not identical with it. The neuraminidase antigen of this Hswl subtype was closely related to that of A/turkey/Ontario/6118/68 and was shown to be Nav4 subtype. After experimental infection of 5-week-old SPF-chickens with the isolate, virus was recovered from various organs including the brain, despite the absence of signs of disease. PMID- 6270507 TI - Effects of conjugated equine oestrogens and oestriol on blood clotting, plasma lipids and endometrial proliferation in post-menopausal women. AB - The effects on blood-clotting factors, plasma lipids and endometrial proliferation of two types of oral oestrogen therapy (a) cyclical therapy with 0.625 mg/day conjugated equine oestrogens for 3 wk followed by 1 wk without treatment, and (b) continuous therapy with 2 mg/day oestriol) were investigated in a 5-mth study involving 22 post-menopausal women. Assays were performed in order to measure blood-clotting factors (activity of the factor II-VII-X complex, anti-thrombin III, euglobulin lysis time) and plasma lipids (total cholesterol, triglycerides, lipoprotein electrophoresis) before treatment and after 3 and 4 mth of treatment. The only significant change found to have occurred was a decrease in the beta: alpha lipoproteins ratio; this was greater in the group treated with conjugated equine oestrogens, but was also statistically significant in the group treated with oestriol. Endometrial proliferation was investigated indirectly by means of the medroxyprogesterone acetate (MPA) test before treatment and after 5 mth of treatment. The response to MPA at the end of 5 mth of treatment in patients previously unresponsive, showed endometrial proliferation to have occurred during treatment with conjugated equine oestrogens but not during treatment with oestriol. PMID- 6270509 TI - Phenotypic change of an SV40-transformed mouse macrophage line, BB-W-531-2 induced by different cultural methods. AB - A simian virus 40 (SV40)-transformed macrophage clone which was established from BALB/cAnN mouse bone marrow cells was used to study the effect of different cultural conditions on the expression of macrophage properties. The macrophage clone, BB-W-531-2 line, expressed and maintained the macrophage properties, immune phagocytosis of Fc- and complement receptors, under the growth-inhibiting conditions of confluent density and of cultivation on bacteriologic dishes with reduced adhesiveness. However, the cells lost their ability to express the macrophage properties dependent upon cell density after repeated culture splits in the growing phase. These cells regained that ability when they were cocultured with cells having macrophage properties. These results suggest that there is a possible correlation between reduced multiplication and the expression of macrophage properties, and that macrophage properties which have been suppressed or blocked may be induced by diffusible factor(s) produced by macrophages. PMID- 6270510 TI - Differentiation of herpes simplex virus type 1 and type 2 by intranuclear tubular structures in clinical infection. PMID- 6270512 TI - On the chemical and structural basis of membrane transducer and receptor systems. AB - Evolution was based initially on polynucleotide systems. From these there developed the polynucleotide-based protein biosynthetic system. Membrane transducers are required by the most primitive cells and must have appeared at a very early stage. They would therefore also be expected to involve polynucleotides. There is substantial biochemical evidence for polynucleotide involvement in membrane transducers. There does not appear to be any reasonable alternative. Many biologically active substances show good matching to a polynucleotide receptor site that gives a good account of receptor function. There appears then to be no doubt that polynucleotides provide the basis of membrane transducers. PMID- 6270511 TI - Maturity-onset diabetes mellitus--toward a physiological appropriate management. AB - "Eumetabolic" therapies of diabetes are defined as those which promote and potentiate a normal physiological pattern of insulin activity--in contrast to the less subtle effects of exogenous insulin or sulfonylureas. Effective eumetabolic therapy would provide better metabolic control and reduce the risk of secondary complications, would be more convenient to administer, and would be especially appropriate for preventive use in the early stages of diabetes. Possible components of a eumetabolic therapy include: aspirin, as a potentiator of glucose stimulated insulin secretion; GTF, to directly enhance the efficacy of insulin; weight loss, exercise, and fasting, to help reduce tissue resistance to insulin; mitochondrial "metavitamins", to optimize the oxidative disposal of excess substrate; a high-fiber, low-fat diet, which appears superior to traditional diabetic diets as a promoter of glucose tolerance. Following a prolonged fast, obese diabetics show substantial improvement in most parameters of insulin function--an effect which is to some degree independent of weight loss; long-term remission of diabetes may be possible if the benefits of therapeutic fasting are conserved by appropriate eumetabolic measures. PMID- 6270513 TI - Coronary artery vasospasm and emotional stress: a hypothetical link to atherosclerosis. AB - A large body of evidence suggests that psychosocial behavior may contribute to the formation of coronary heart disease. It is proposed that emotionally-induced coronary vasospasm leads to the development of coronary atherosclerosis. This hypothesis is based on two observations: a) coronary vasospasm is present at the site of organic lesions and may be antecedent to the development of the atherosclerotic plaque, and b) emotional stress results in an alpha adrenergically mediated increase in coronary vascular resistance. PMID- 6270514 TI - A biochemical basis for the actions of lithium on behaviour and on immunity: relapsing and remitting disorders of inflammation and immunity such as multiple sclerosis or recurrent herpes as manic-depression of the immune system. PMID- 6270515 TI - Neurotoxicity of folates: implications for vitamin B12 deficiency and Huntington's chorea. AB - Recent work has shown that several folates interact with excitatory kainic acid receptors in the mammalian brain and appear to have agonist activity at these receptors. Since kainic acid is a potent neurotoxin it is possible that folates share this toxicity and that high levels of folates result in neuronal damage. Levels of methyltetrahydrofolate are markedly elevated in vitamin B12 deficiency, a disease associated with neuronal destruction. We propose that this destruction occurs as a result of a neurotoxic action of methyltetrahydrofolate. Injection of kainic acid into the basal ganglia of experimental animals produces a pattern of damage similar to that found in patients dying of Huntington's chorea. It is possible that the underlying defect in this disease resides in the pathways of folate metabolism such that a neurotoxic excess of folates accumulates in the central nervous system. Such a disease process might be arrested by antifolate drugs. PMID- 6270516 TI - Hypothesis: the bronchial Kulchitsky (K) cell as a source of humoral biologic activity. AB - The bronchial Kulchitsky cells are scattered specific cells which lie close to the basement membrane of the bronchi and bronchioles. Electron microscopy reveals that they contain electron-dense granules similar to that seen in cells with a known endocrine function. In addition, experimentally induced degranulation of the K cell suggestive of a secretory process, and the likelihood that these cells are precursors of small cell lung carcinoma (which often elaborates humoral substances) has led to the postulate that this bronchial cell serves a humoral role of either a paracrine or endocrine nature. We have found that the bronchial K cell of man contains a calcitonin-like polypeptide which, immunologically and chemically, is not dissimilar to the hormone produced by the C cells of human thyroid. This finding may help explain the persistence of serum immunoreactive calcitonin (iCT) after total thyroidectomy, the fact that thyroidectomized man does not manifest any profound alteration of calcium metabolism, and why small cell cancer of the lung is frequently associated with hypercalcitonemia. In addition, the finding of K cell hyperplasia in chronic bronchitis and emphysema may explain the occurrence of hypercalcitonemia in patients with these diseases and some lung cancers of cell types other than the small cell variety. Further studies are needed to elucidate the role of K cell iCT, and to determine what other hormones might also be elaborated by this diffuse system of bronchial cells. PMID- 6270517 TI - Rabbit behavioral model of marijuana psychoactivity in humans. AB - In a genetically unique colony of tetrahydrocannabinol-seizure susceptible (THC SS) rabbits, nonfatal convulsions are elicited by delta 9THC, the major psychoactive ingredient of marijuana. The major characteristics of cannabinoid produced psychoactivity (the "high") in humans, e.g., dose-effect relationships, specificity of response to only psychoactive cannabinoids, tolerance development, EEG correlates, and delta 9THC-cannabidiol interactive effects, are also characteristics of cannabinoid-induced behavioral convulsions in the rabbits. Because of these and other theoretical and practical considerations, it is hypothesized that the THC-SS rabbit represents a novel laboratory animal model of marijuana-induced psychoactivity in humans. PMID- 6270518 TI - On the molecular structure of the ion channel associated with the acetylcholine receptor: a beta-barrel? PMID- 6270519 TI - Probable mechanism for the loss of Barr body in human female tumor with special reference to breast cancer. AB - One of the X-chromosomes by a random inactivation process condenses to form X chromatin (Barr body) in early embryonic life. Once this occurs, it is final and fixed for that cell and all its descendants (1,2). However, numerous investigators have observed extreme variations in Barr body frequency in tumour cells. For example, Sohval and Gains (3) reported an absence of the characteristic Barr body pattern of 19 of the 27 teratomas from females and Moore and Barr (4) observed significant variation in Barr body counts in breast cancer cells. Furthermore, a good correlation was made between the frequency of Barr body and prognosis (5,6,7,8,9). From a retrospective study, it was shown that tumours with low Barr body frequency (BBF) had a significant correlation with blood vessel invasion (BVI) and poor prognosis (10). But the reason why patients with low BBF in the tumor with BVI get early recurrence is not known. In this paper an attempt has been made to suggest a mechanism which may be involved in reducing the BBF with high malignant potentiality. PMID- 6270520 TI - Polymyalgia rheumatica and giant cell arteritis--rational diagnosis and treatment predicated and disordered prostaglandin metabolism. AB - We suggest that polymyalgia rheumatica with giant cell arteritis (PR-GCA) is an arachidonic acid metabolites mediated disease which can be diagnosed more accurately and monitored more precisely for therapeutic benefits by the serial determinations of the major urinary prostaglandin F, serum urinary lysozymes, serum acid phosphatase, and serum angiotensin converting enzyme rather than by the erythrocyte sedimentation rate, and, when necessary by temporal artery biopsy. The pathogenetic role proposed for prostaglandins (PG) and, even more precisely perhaps, the leukotrienes in this disease is consistent with the several published clinical observations that non-steroidal anti-inflammatory drug treatment produces in some cases a therapeutic paradox of symptomatic relief with concurrent, if clinically silent, progression of the arteritis, even to blindness. Furthermore, the impressive response of PR-GCA to low maintenance dose steroid therapy, a clinical conundrum for decades, is rationally explained on the basis of depressed or obstructed PG metabolism early on in the metabolic cascade. These views warrant clinical evaluation, confirmation or correction in whole or in part, and may increase our understanding of PR-GCA. PMID- 6270521 TI - Loss of delta-6-desaturase activity as a key factor in aging. AB - Aging is characterized by a wide variety of defects, particularly in the cardiovascular and immune systems. Cyclic AMP levels fall, especially in lymphocytes. Delta-6-desaturase (D6D) levels have been found to fall rapidly in the testes and more slowly in the liver in aging rats. D6D is an enzyme which converts cis-linoleic acid to gamma-linolenic acid (GLA). Other factors which inhibit D6D activity are diabetes, alcohol and radiation, all of which may be associated with accelerated aging. In meat eaters or omnivores which can acquire arachidonic acid from food, the main consequences of D6D loss will be deficiencies of GLA, dihomogamma-linolenic acid (DGLA) and prostaglandin (PG) E1. PGE1 activates T lymphocytes, inhibits smooth muscle proliferation and thrombosis, is important in gonadal function and raises cyclic AMP levels in many tissues. It is a good candidate for a key factor lost in aging. Moderate food restriction, the only manoeuvre which consistently slows aging in homoiotherms, raises D6D activity by 300%. Other factors important in regulating D6D and the conversion of GLA to PGE1 are zinc, pyridoxine, ascorbic acid, the pineal hormone, melatonin, and possibly vitamin B3. GLA administration to humans has been found to lower blood pressure and cholesterol, and to cause clinical improvement in patients with Sjogren's syndrome, scleroderma and alcoholism. These diseases are associated with some features of accelerated aging. The proposition that D6D loss is not only a marker of aging but a cause of some of its major manifestations is amenable to experimental test even in humans. The blocked enzyme can be by-passed by giving GLA directly. PMID- 6270522 TI - Isolation of enterovirus 70 from patient with acute hemorrhagic conjunctivitis-- Key West, Florida. PMID- 6270523 TI - Trends in veterinary immunology. Developments in immunoprophylaxis. PMID- 6270524 TI - Diagnosis of feline leukemia virus infections. PMID- 6270525 TI - Rota-coronavirus vaccination of pregnant cows. PMID- 6270526 TI - Interactions of ligands with morphine and enkephalin receptors are differentially affected by guanine nucleotide. PMID- 6270527 TI - The interaction of ervatamine and epiervatamine with the action potential Na+ ionophore. PMID- 6270528 TI - Calcium-dependent blockade of cardiac cyclic AMP accumulation by batrachotoxin and veratridine. PMID- 6270529 TI - Studies on the relationship of gamma-aminobutyric acid-stimulated diazepam binding and the gamma-aminobutyric acid receptor. PMID- 6270530 TI - Ornithine decarboxylase induction in liver- and hepatoma-derived cell cultures: no detectable differences between control and 3-methylcholanthrene-treated cells. PMID- 6270532 TI - Xanthine derivatives that selectively inhibit cyclic GMP hydrolysis potentiate cardiac contractile effects of isoproterenol but not those of bethanecol. PMID- 6270531 TI - Selective phosphodiesterase inhibition and alterations of cardiac function by alkylated xanthines. PMID- 6270534 TI - The effect of alprenolol on the beta-receptor and adenylate cyclase activity in rabbit heart membranes. PMID- 6270533 TI - Metabolism of 9-beta-D-xylofuranosyladenine by the Chinese hamster ovary cell. PMID- 6270535 TI - Differential affinities of 5-(2-halogenovinyl)-2'-deoxyuridines for deoxythymidine kinases of various origins. PMID- 6270536 TI - Stereochemical considerations in the binding of nonsteroidal estrogens to the estrogen receptor. PMID- 6270537 TI - Regulation of myometrial adrenoreceptors and adrenergic response by sex steroids. PMID- 6270538 TI - Cholera toxin-stimulated cyclic AMP accumulation in glial tumor cells: modulation by Ca2+. PMID- 6270539 TI - N-demethylchlordimeform: a potent partial agonist of octopamine-sensitive adenylate cyclase. PMID- 6270541 TI - Irreversible inactivation of the opiate receptors in the neuroblastoma x glioma hybrid NG108-15 by chlornaltrexamine. PMID- 6270540 TI - Recognition of insulin-releasing fuels by pancreatic B-cells: alpha ketoisocaproic acid is an appropriate model compound to study the role of B-cell metabolism. PMID- 6270542 TI - Quinidine interactions with Myxicola giant axons. PMID- 6270543 TI - Glutamate and aspartate agonists structurally related to ibotenic acid. AB - This mini-review describes a noval class of excitatory heterocyclic amino acid. The selective interactions of these synthetic amino acids with the central glutamic acid (GLU) and aspartic acid (ASP) receptors have been established on the basis of microelectrophoretic techniques using glutamic acid diethyl ester (GDEE) and alpha-aminoadipic acid (alpha-AA) as selective antagonists for GLU and ASP, respectively. The parent compound, ibotenic acid (IBO) preferentially activates ASP receptors, but elongation of the side chain of IBO afforded homoibotenic acid (homo-IBO), a GLU agonist. The introduction of bulky substituents into the heterocyclic ring of homo-IBO resulted in a dramatic increase in potency. Alteration of the position of the side chain in IBO to give alpha-amino-5-methyl-3-hydroxy-4-isoxazoleacetic acid (AMAA), preserved the ASP agonism. However, elongation of the side chain of AMAA gave alpha-amino-5-methyl 3-hydroxy-4-isoxazolepropionic acid (AMPA), which is a very powerful neuronal excitant with selective interaction with the GLU receptors. None of the new compounds are inhibitors of the binding of 3H-kainic acid (3H-KAIN) to rat brain membranes, indicating that the mechanism of action of these compounds is different from that of the neurotoxic compound KAIN. The described compounds may be important tools in future investigations of the physiological role and the mechanism of action of ASP and GLU in the central nervous system. PMID- 6270544 TI - GABA agonists. Development and interactions with the GABA receptor complex. AB - This review describes the development of GABA receptor agonists with no detectable affinity for other recognition sites in GABA-mediated synapses. The key compounds are THIP, isoguvacine, and piperidine-4-sulphonic acid (P4S), developed via extensive structural modifications of the potent but not strictly specific GABA agonist muscimol. The structural parameters, which have to be considered in the design of GABA agonists are discussed on the basis of the structures and biological activities of these GABA agonists and a number of related compounds. A model, which summarizes our present knowledge of the structure of the postsynaptic GABA receptors complex, is presented, and the interaction of GABA agonists with various sites in this complex is discussed. Of particular interest are the effects of GABA agonists on the binding of diazepam to the benzodiazepine binding site, assumed to be a structural unit of the GABA receptor complex. While rigid molecules like THIP are capable of activating the GABA receptors, a certain degree of conformational mobility of GABA agonists apparently is a prerequisite for stimulation of diazepam binding in vitro at 0 degree C. The findings suggest that GABA receptor functions involve conformational changes of certain elements, including the attempts to develop GABA agonists with desirable pharmacokinetic and toxicological characteristics. While muscimol is a toxic compound, THIP is well tolerated by animals, and in contrast to isoguvacine, THIP penetrates into the brain after systemic administration to animals, a difference which can be explained on the basis of their protolytic properties. The attempts to develop pro-drugs of isoguvacine capable of penetrating the blood-brain barrier with subsequent decomposition in the brain tissue to isoguvacine are described. PMID- 6270546 TI - Hippocampal glutamate receptors. AB - For years, the hippocampus has been the privileged domain of anatomists and electrophysiologists for investigating various neurobiological processes. The present review deals with recent work which shows that this structure is also well suited to study the role of glutamate as a neurotransmitter and more particularly the characteristics of glutamate receptors and their possible involvement in hippocampal function. After a brief description of the main anatomical features of the hippocampus, we attempt a critical evaluation of the electrophysiological studies of hippocampal glutamate receptors. We then describe the properties of Na-independent 3H-glutamate binding sites in hippocampal membranes, and discuss the possibility that these binding sites are related to postsynaptic glutamate receptors. Finally we show that these binding sites are extremely labile and that hippocampal membranes possess various mechanisms which regulate their number. In particular we develop the idea that the calcium stimulation of 3H-glutamate binding in hippocampal membranes may be the mechanism by which electrical activity regulates the number of glutamate receptors at hippocampal synapses and thus induces long-lasting changes in synaptic transmission. PMID- 6270548 TI - Protein kinase activity tightly bound to liver polysomes. AB - Rat liver polysomes washed with 0.5-1.5 M KCl at 37 degrees C keep a constant protein kinase activity revealed only by auto-phosphorylation of ribosomal proteins. The enzyme catalyzes the transfer of the gamma-phosphate group from ATP to serine (75%) but also to threonine residues (25%). It is released when polysomes are dissociated into subunits using centrifugation through a sucrose gradient containing a high K+/Mg++ ratio. Its properties have been compared with those of the two other enzymatic activities which are, in contrast, washed out during salt treatment of polysomes. After release upon polysome dissociation, this third activity is able to phosphorylate histone II A. Protection of the enzyme in the polysome structure against salt treatment, suggests that it is located at the junction of the two subunits. PMID- 6270547 TI - Protein synthesis in artemia salina. Eucaryotic elongation factor eEF-Ts is a transphosphorylase. AB - It is thought that eucaryotic elongation factor eEF-Ts catalyzes the replacement of GDP for GTP on eucaryotic elongation factor eEF-Tu. We have found that eEF-Ts displays a strong nucleoside diphosphate phosphotransferase activity. This transferase activity resides in a dimer molecule of a subunit molecular weight close to 30,000. The transfosforylating activity of eEF-Ts results in a stimulatory effect of ATP, GTP, UTP and CTP on protein synthesis provided that GDP is present. The specificity for guanine nucleotides in protein synthesis resides only in eEF-Tu. PMID- 6270545 TI - Pharmacology of GABA-mediated inhibition of spinal cord neurons in vivo and in primary dissociated cell culture. AB - In this paper it is shown that the postsynaptic GABA-receptor chloride ion channel complex is composed of several functional subunits. There are probably at least two stereospecific locations on the receptor for GABA-binding and both must be occupied to obtain an increase in chloride conductance. The interaction between these sites is uncertain but there could be either positive cooperativity between the sites or only a requirement that both sites are occupied without occupation of either site affecting the affinity for GABA of the other site. There is a chloride conductance channel coupled to the GABA receptor which opens for an average of 20 msec and has an average conductance of 18 pS. The GABA coupled chloride channel may or may not have the same composition as the glycine coupled chloride channel. In addition to the GABA-recognition site and the chloride ion channel, GABA-receptors must have additional binding sites or modulator sites where drugs can bind to modify GABA activation of the GABA receptor. The convulsant PICRO binds to a site which is independent of the GABA site and PICRO reduces GABA responses. Barbiturates and benzodiazepines augment GABA-responses without reducing GABA-binding and thus they must bind to a modulator site independent of the GABA recognition site. Whether or not this is the same site as the PICRO binding site is uncertain. Thus, the GABA-receptor chloride ion channel complex is composed of at least: 1) two GABA-binding sites; 2) a chloride ion channel; 3) a convulsant binding site (PICRO-binding site) and 4) an anticonvulsant binding site. This organization serves several obvious purposes. First, since two GABA-molecules are required to activate GABA-coupled chloride ion channels, the dose-response relationship for GABA is sigmoidal and steep. Thus minor shifts in GABA affinity will produce large alterations in GABA responses and the GABA receptor can be easily modulated. Second, since the receptors has binding sites for convulsant and anticonvulsant compounds which decrease and increase GABA-responses, GABAergic inhibition can easily be modulated. PMID- 6270550 TI - The activity of ketoconazole in clinical isolates of candida albicans cultured in a mycelium promoting medium. PMID- 6270549 TI - Membrane defects in paramyotonia congenita with and without myotonia in a warm environment. AB - Three patients with paramyotonia congenita and 3 control persons were biopsied for an in vitro investigation of the sarcolemmal membrane parameters and of the contractile properties of paramyotonic muscle. At 37 degrees C, paramyotonic muscle fibers had normal resting potentials, but on cooling to 27 degrees C they depolarized. Depolarization to -60 mV caused spontaneous activity, and further depolarization to -40 mV caused inexcitability. Depolarization could be prevented by the application of tetrodotoxin, a finding suggesting a defect in the Na channels. Analysis of the membrane current densities using voltage clamps with 3 microelectrodes revealed that in paramyotonic patients at 37 degrees C all component conductances were normal, except for a decreased Cl conductance in the patient who had myotonia in a warm environment. At 27 degrees C, the Na and Cl conductances were abnormally high. The K conductance was always normal. The results explain the clinical symptoms of weakness and paralysis. Potassium- and caffeine-contracture experiments gave normal results. The clinical symptom of paramyotonic stiffness, therefore, has not been explained by these studies. PMID- 6270551 TI - Coxsackie B viruses and autoimmune diabetes. PMID- 6270552 TI - Autonomic abnormalities and autoantibodies to beta-adrenergic receptors. AB - We identified autoantibodies to beta 2-adrenergic receptors in the plasma of three apparently normal subjects, four patients with allergic asthma, one subject who was "preallergic" (at risk of allergy), and one patient with cystic fibrosis. Although these antibodies appeared to be heterogeneous, they shared the ability to affect binding of [125]protein A to calf-lung membranes, to inhibit beta adrenergic ligand binding to calf-lung bet-adrenergic receptors, and to precipitate solubilized calf-lung beta-adrenergic receptors in an indirect immunoprecipitation assay. The presence of autoantibodies to beta-adrenergic receptors in these subjects correlates with abnormal autonomic responsiveness characterized by alpha-adrenergic and cholinergic hypersensitivity and beta adrenergic hyposensitivity. These findings suggest that autoantibodies to beta adrenergic receptors may play a part in the development of ment of autonomic abnormalities. PMID- 6270554 TI - Autoantibodies and beta-adrenergic receptors. PMID- 6270553 TI - Preliminary observations of the effects on breast adenocarcinoma of plasma perfused over immobilized protein A. PMID- 6270555 TI - Does perfusion with treated plasma cure cancer? PMID- 6270556 TI - Defective EBV-specific suppressor T-cell function in rheumatoid arthritis. AB - Several lines of evidence, including high antibody titers to Epstein-Barr virus (EBV)-associated antigens and rapid transformation of B cells into lymphoblastoid cells lines, suggest an association between EBV and rheumatoid arthritis. When lymphocytes from normal immune donors were infected with EBV in culture, they produced an exponentially increasing number of immunoglobulin-secreting cells for eight to 10 days. Thereafter, there was a marked late suppression of their response, mediated by immunoregulatory T cells; by 12 days in culture, this suppression averaged 90 per cent. Lymphocytes from 20 EBV-immune patients with rheumatoid arthritis also responded with increasing production of immunoglobulin secreting cells, but the late suppression expected in immune donors was absent. Tests of several other T-cell functions in these patients gave normal results, suggesting a more restricted defect in suppressor-T cell function relating specifically to EBV. Since EBV persists in host B cells and thus represents a potential stimulus for immunoglobulin production, this persistence, along with a specific regulatory T-cell defect, may contribute to many of the immune abnormalities underlying rheumatoid arthritis. PMID- 6270558 TI - Erythrocyte sodium-potassium-ATPase and sodium transport in obesity. PMID- 6270557 TI - A cyproheptadine-reversible defect in ACTH control persisting after removal of the pituitary tumor in Cushing's disease. AB - We studied two phases of cortisol feedback suppression of ACTH in nine patients who had had adrenalectomy for Cushing's disease. Four had been treated by adrenalectomy alone and presumably had ACTH-secreting pituitary tumors. Five others were studied two or more years after transsphenoidal removal of an ACTH secreting microadenoma. In both groups, cortisol-ACTH feedback during the first 30 minutes of cortisol infusion was abnormal; plasma ACTH fell only 2.7 +/- 2.6 per cent (mean +/- S.E.), as compared with 28.0 +/0 10.1 per cent in five hypoadrenal controls (p less than 0.01). The fall in ACTH during the second phase of cortisol infusion was similar in the patients and the controls. Cyproheptadine corrected the feedback abnormality occurring during the first phase in both groups of patients with Cushing's disease; ACTH fell by 24.4 +/- 4.8 per cent (P less than 0.005). Persistence of a cortisol-ACTH feedback abnormally after removal of the pituitary tumor in Cushing's disease, as well as the correction by cyproheptadine, suggests that higher centers have a role in the pathophysiology of Cushing's disease. PMID- 6270559 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercise. Case 47-1981. Fatal cerebral disorder after removal of parotid tumor. PMID- 6270560 TI - Deficiency of T helper cells in transient hypogammaglobulinemia of infancy. AB - We studied 17 patients with transient hypogammaglobulinemia of infancy to define the immunologic defect responsible for this disorder. The number of circulating B cells in these patients was normal, as was the ability of the B cells to synthesize immunoglobulins when stimulated with Epstein-Barr virus, a direct B cell activator. However, the capacity of the B cells to synthesize IgG in response to pokeweed mitogen, a T-cell-dependent B-cell activator, was depressed. Experiments with cultured lymphocytes indicated that excess suppressor-cell activity was not present in these patients, but that their T cells were deficient in providing help to B cells from their normal parents. A numerical deficiency in T4-positive (T4+) helper cells was found. Patients who had recovered from the disorder had a normal number of T4+ helper cells. Our results indicate that a numerical, as well as a functional, deficiency in helper T cells underlies the deficiency in IgG production in transient hypogammaglobulinemia of infancy. PMID- 6270562 TI - Revival of neuronal electrophysiology. PMID- 6270563 TI - Making potassium run uphill. PMID- 6270561 TI - A variant of chronic granulomatous disease: deficient oxidative metabolism due to a low-affinity NADPH oxidase. PMID- 6270564 TI - Passing messages downstream. PMID- 6270565 TI - Calcium helps to send out signals. PMID- 6270566 TI - Towards understanding the acetylcholine receptor. PMID- 6270567 TI - Demethylation of CpG sites in DNA of early rabbit trophoblast. PMID- 6270568 TI - Cell-surface expression of influenza haemagglutinin from a cloned DNA copy of the RNA gene. AB - By replacing either the eight early or the late genes of SV40 with a cloned copy of the influenza virus haemagglutinin gene we have constructed recombinant viruses which, in infected cells, express large quantities of haemagglutinin. This glycoprotein, over 10(8) molecules of which are produced per cell, is identical in molecular weight to authentic influenza virus haemagglutinin, accumulates at the cell surface and displays haemabsorbing activity. PMID- 6270569 TI - Insertion of DNA activates the cryptic bgl operon in E. coli K12. PMID- 6270570 TI - Analysis of the terminus region of the Bacillus subtilis chromosome. PMID- 6270571 TI - Autoregulation of androgen production in a primary culture of rat testicular cells. PMID- 6270572 TI - Induction of ovulation in vitro by LH and catecholamines in hens is mediated by alpha-adrenergic receptors. PMID- 6270573 TI - Potassium depletion decreases the number of 3H-ouabain binding sites and the active Na-K transport in skeletal muscle. PMID- 6270574 TI - Persistence of non-integrated viral DNA in bovine cells transformed in vitro by bovine papillomavirus type 2. PMID- 6270575 TI - Enkephalin as a transmitter for presynaptic inhibition in sympathetic ganglia. PMID- 6270576 TI - Transcriptional regulation of the prolactin gene by ergocryptine and cyclic AMP. PMID- 6270578 TI - Demonstration of a syncarcinogenic effect of endogenous MuLV, rescued from MNU induced leukemias, and suboptimal MNU doses in mice. AB - It was demonstrated that the combined action of activated endogenous MuLV isolated from MNU-induced leukemias and suboptimal doses of MNU yields a significantly high incidence of leukemias and tumors. Treatment with one agent alone shows low effects only. The findings were discussed in view of the mechanisms in which viruses and chemical carcinogens interact in a syncarcinogenic processes. PMID- 6270577 TI - [Urachal carcinoma]. PMID- 6270579 TI - Establishment and characterization of permanent cell lines from patients with acute and hairy cell leukemia. AB - Four cell lines were established from peripheral blood of patients with leukemia. All lines express the Epstein--Barr virus nuclear antigen (EBNA) and therefore should be classified as lymphoblastoid cell lines. However, one of the lines UHKT 5 established from a patient with acute myelomonocytic leukemia has some features not typical for lymphoblastoid cell lines. The cells resemble to macrophages with the phagocytic ability for yeasts, strong alpha-naphthylacetate esterase positivity in phagocytizing cells and unusually long villi. Another line UHKT-7 established from a patient with hairy cell leukemia expresses the same isotype of membrane immunoglobulins as the original hairy cells. PMID- 6270580 TI - In vitro studies of a co-carcinogenic effect of vaccinia and herpes group viruses. AB - The results of studies of a co-carcinogenic effect of two human infectious viruses in tissue culture are reported here. Viable vaccinia virus actively replicating in the cells of primary BALB/c tissue culture and in a number of continuous murine cell lines has been shown to induce in them expression of major structural p30 protein of murine retroviruses. Vaccinia virus has been also shown to cause biochemical transformation of murine cells. Evidence for the capacity of herpes simplex virus type 2 to induce malignant transformation of BALB/3T3 murine cell line has been obtained and confirmed by transplantation to mice. Transformed cell clones did not contain complete infectious herpes simplex virus but were resistant to superinfection with this virus. N-tropic endogenous murine retrovirus of C type with buoyant density in the saccharose density gradient of 1.18 g/cm3 and a reverse transcriptase activity was expressed in the transformed cells. In the virion structure six proteins typical of these viruses with the prevalence of p30 have been demonstrated. Competitive radioimmunoassay revealed a very high level of virus production: p30 level reached 7500 ng p30 R-MuLV per mg of viral protein. Specificity of this results was shown in control experiments. PMID- 6270581 TI - The expression of virus-specific nucleotide sequences in Rous sarcoma cells. AB - Poly(A)-enriched virus-specific mRNAs (v-mRNA) are revealed in the subcellular fractions (nuclei, mitochondria, polyribosomes and cytosol) from chicken and hamster Rous sarcoma cells. The qualitative and quantitative differences in v mRNA populations from permissive and non-permissive tumors are shown. The sedimentation analysis of v-mRNAs allowed to ascertain the presence of three molecular classes of v-mRNA in nuclei and cytoplasm of chicken Rous sarcoma cells (35S, 28S and 22S, and 35S, 26-28S and 20S, respectively). While two size classes (33S and 20S) are shown in hamster Rous sarcoma nuclei, only 24S v-mRNA is revealed in the cytoplasm of its tumor tissue. Thus, the oncovirus gene expression is restricted in the non-permissive tumor cells, unlike of the permissive chicken Rous sarcoma cells, both in the level of transcription, and in the processing and transfer v-mRNAs from nucleus to cytoplasm. The virus gene function peculiarities in the tumors of birds and mammals are discussed. PMID- 6270582 TI - Calculation of reactivity indices for benz(a)anthracene and benzo(a)pyrene and their approximative models of complex with enzyme epoxidase. AB - By theoretical methods of quantum mechanics an interaction between carcinogenic hydrocarbons -- benz(a)anthracene, benzo(a)pyrene -- and various groups simulating an effect of enzyme oxidase has been studied. If accord is to be achieved with formation of epoxides of these compounds in vivo according to the bay region theory (i. e. in A region) an interaction between the enzyme involving an electron-acceptor group and the bay region of hydrocarbon under simultaneous steric hindrance of the K region is necessary. Another possibility of obtaining the highest reactivity in the A region is to assume the formation of the radical cation as a first intermediate in the interaction with enzyme. PMID- 6270583 TI - Activity of the pituitary-adrenal system in rat fetuses subjected to encephalectomy in early or late stages of pregnancy. AB - The corticostimulating activity of the fetal rat hypophysis was studied on day 21, in fetuses deprived of their hypothalami by total removal of the brain (encephalectomy) in early (day 16) or late (day 19) stages of gestation. On day 19, the fetal manipulation was performed either on intact or adrenalectomized dams. The data show that: (1) Compared to controls, encephalectomy does not alter pituitary ACTH content in fetuses of intact mothers but that pituitary ACTH content does not significantly increase in encephalectomized fetuses of adrenalectomized mothers whereas it does in the littermate control fetuses. (2) Compared to controls, circulating ACTH levels are reduced by encephalectomy on both day 16 and day 19; an increase in ACTH levels occurs in both fetal groups in response to maternal adrenalectomy. Decapitation removes this fetal source of ACTH. (3) Adrenal glands are not as much atrophied by encephalectomy as by decapitation on day 16, whereas adrenal weight is decreased to the same extent by encephalectomy and by decapitation on day 19. However, the marked increase in circulating ACTH in both control and encephalectomized fetuses caused by maternal adrenalectomy is probably the cause of the significant increase in adrenal weight in these groups. (4) Adrenal corticosterone content in all the groups but not always adrenal weight, is a fairly accurate reflection of the ACTH levels measured. The results suggest that: (1) Pituitary ACTH content is not a sensitive measure of dynamic changes in synthesis and secretion in rat fetuses. (2) Changes in basal plasma ACTH in response to fetal encephalectomy show the presence of both an autonomous corticotropic function and a cephalic regulation of the corticotropic function from day 16 of fetal life and beyond. Pituitary autonomy seems higher in earlier than in later stages of gestation. (3) There is feedback of corticosterone into the pituitary of fetal rats; a marked increase in ACTH release is caused by the removal of the maternal corticosteroids. PMID- 6270585 TI - Effect of intraventricular beta-endorphin and morphine on hypothalamic-pituitary adrenal activity and the release of pituitary beta-endorphin. AB - The effects of intraventricular (i.v.t.) morphine sulfate (MS) and beta-endorphin (beta-EP) on pituitary-adrenal activity and the release of pituitary beta-EP were studied in rats. Pituitary-adrenal activity was monitored by measuring plasma corticosterone (CS) levels. 45 min after i.v.t. injection, both MS and beta-EP caused dose-related increases in plasma CS, with beta-EP being approximately ten times more potent on a molar basis. MS injected i.v.t. at 0.3, 1.0, 3.0 and 10.0 microgram did not cause a significant reduction in pituitary immunoreactive (i.r.) beta-EP, but did cause an increase in plasma i.r. beta-EP at 3 microgram of MS. beta-EP injected i.v.t. at 1.5 microgram caused a reduction of pituitary i.r. beta-EP. Since i.v.t.-injected beta-EP may have contributed to the measured plasma i.r. beta-EP, a nonimmunoreactive analog (Des-Asn20-beta c-EP) was used to assess the change in plasma i.r. beta-EP. 5 microgram of DES-Asn20-beta c-EP injected i.v.t. caused increases in plasma i.r. beta-EP and CS, as well as a 40% reduction in pituitary i.r. beta-EP. The concomitant intraperitoneal (i.p.) injection of naloxone HCl (10 mg/kg) significantly blocked the increase in plasma CS induced by 5 microgram of beta-EP. When naloxone HCl, 10 mg/kg was injected alone, a significant increase in plasma CS was found. The results indicate that i.v.t. beta-EP is more potent than MS in causing the release of pituitary ACTH and beta-EP. These findings are consistent with a role for brain endorphins in the regulation of CRF release. PMID- 6270584 TI - Plasma norepinephrine, epinephrine and dopamine responses to intracerebral administration of a met-enkephalin analog, D-ala2-met-enkephalinamide, in rats. AB - Intracisternal (i.c.) administration (25 microgram or 42.6 nmol) of the potent met-enkephalin analog, D-ala2-met-enkephalinamide (DALA) in adult male rats increased plasma concentrations of norepinephrine and epinephrine for more than 90 min. Intra-arterial (i.a.) administration of the same or a fourfold larger dose was without any effect. I.c. administration of human beta-endorphin (25 microgram or 7.25 nmol) produced much greater plasma responses of all three catecholamines than seen with DALA. An i.a. dose of naloxone, which had been shown previously to inhibit the plasma catecholamine responses to beta-endorphin, failed to inhibit plasma catecholamine responses to DALA. Also, naloxone alone failed to alter the basal plasma concentrations of the three catecholamines. The data are consistent with an effect of DALA acting at a presently unknown brain site to increase central sympathetic outflow, thus increasing plasma concentrations of norepinephrine and epinephrine. It appears probable that this effect of DALA is mediated at a different opioid receptor and perhaps a different brain site than the effect of beta-endorphin to increase central sympathetic outflow. PMID- 6270586 TI - Superior cervical ganglionectomy depresses norepinephrine uptake, increases the density of alpha-adrenoceptor sites, and induces supersensitivity to adrenergic drugs in rat medial basal hypothalamus. PMID- 6270588 TI - A steroid derivative, R 5135, antagonizes the GABA/benzodiazepine receptor interaction. PMID- 6270587 TI - Effect of neonatal treatment with monosodium glutamate on the secretion of alpha MSH, beta-endorphin and ACTH in the rat. AB - Plasma alpha-MSH, beta-endorphin and ACTH were measured at 60 days of age in rats which had been injected during the neonatal period with monosodium glutamate (MSG). Although the arcuate nucleus tuberoinfundibular dopaminergic and cholinergic system was lesioned by the MSG, no change in circulating alpha-MSH, ACTH and corticosterone levels was observed under basal conditions, after ether stress of adrenalectomy. In contrast, a moderate, but significant decrease in plasma beta-endorphin was noticed after MSG treatment. PMID- 6270589 TI - Interactions between cholinergic drugs, gamma-aminobutyric acid and cyclic guanosine monophosphate on picrotoxin-induced convulsive-seizure threshold. PMID- 6270591 TI - [What is meant by minimal carcinoma of the breast and what should be done]. PMID- 6270590 TI - Exsanguination from rupture of an occult hepatoma in a comatose patient: case report. PMID- 6270592 TI - [Solitary metastases of the lung]. PMID- 6270593 TI - [Modern surgical staging of lung cancer]. AB - The estimation of the extension of a lung cancer is actually made according to the rules of TNM system. On the basis of the reports of 100 patients who underwent thoracotomy and were staged according to this system after hystological examination of resected specimen (pTNM), the authors consider some not yet clear aspects of this staging. In particular they underline the wide difference between clinical and post-histological staging; the high rate of nodal involvement, if the surgeon always perform a radical excision of the lymph nodes; the further need of accuracy for the data N2 and T3; the role of the anatomo-pathologist for the correct staging pTNM. PMID- 6270594 TI - [Non-hypoglycemic extra-pancreatic nesidioblastoma. Observations on a case]. PMID- 6270595 TI - [Possible use of dietetic fibers in blood lipid disorders in nephropathic and dialyzed patients]. PMID- 6270596 TI - [Local fixed antigens and experimental nephritis in the rat]. PMID- 6270597 TI - Doubly projecting neurons of the dorsal column nuclei. AB - In a sample of 1700 neurons recorded from the dorsal column nuclei of the cat, 44 (2.6%) were found to send an axon down the dorsal spinal cord. Fully 70% of these caudally projecting neurons also sent an axon to the ventral thalamus. Nearly all had small cutaneous receptive fields distally on the forelimb and displayed response properties similar to other neurons of the dorsal column nuclei. Most were isolated along the lateral and medial margins of the cuneate nucleus rostral to the obex, and many were excited or inhibited by pericruciate cerebral stimulation. A few clearly were excited monosynaptically from the contralateral cerebral cortex at a latency that required the largest pyramidal tract fibers. These neurons probably comprise an important subset that regulates the flow of sensory information in spinal and brainstem somatic sensory pathways. PMID- 6270598 TI - Nucleus laminaris of the torus semicircularis: projection to spinal cord in reptiles. AB - Neurons in nucleus laminaris of the torus semicircularis were retrogradely labeled following application of horseradish peroxidase (HRP) to the cervical spinal cord in two lizards (Gekko gecko and Iguana iguana) and a turtle (Pseudemys scripta). Different patterns of cell labeling were seen among the species studied and may be related to the distinctive differences in head and body movements seen in these animals during defensive, aggressive and social behaviors. PMID- 6270599 TI - Descending projections to the cervical spinal cord in the developing kitten. AB - The distribution of neurons filled by retrograde axonal transport of horseradish peroxidase from the cervical enlargement is described in kittens prior to and following the time of appearance of mature alpha-motoneuron responses to motor cortical stimulation (at 107-111 days gestational age; about 41 days postnatally). Cortex and brainstem reconstructions of the distributions of filled neurons demonstrate a well-defined, discrete projection from cortical area 4 to spinal cord segments C3 to C8, both in mature and immature (20 and 24 days postnatal) animals. In addition, appropriate rubrospinal, reticulospinal and vestibulospinal projections were present at all ages studied. PMID- 6270600 TI - Projection of tooth pulp afferents to the brainstem and to the cortex in the cat. AB - The projection of tooth pulp afferents in the spinal trigeminal nucleus (N.V.sp.) and the effect of dorsolateral medullotomy on cortical potentials evoked by electrical stimulation of Adelta tooth pulp nerve fibres were studied in cats. It was confirmed that antidromic responses were recorded in the tooth pulp nerve to stimulation of the ipsilateral but not the contralateral N.V.sp.[11]. Dorsolateral medullotomy at the level of the obex did not alter the cortical potentials induced by single pulses applied to the tooth pulp. It is concluded that Adelta tooth pulp afferents project exclusively to the ipsilateral trigeminal nucleus and that the impulse transmission to the cortex persists after transection of the pars caudalis of the N.V.sp. PMID- 6270601 TI - Two types of dopaminergic nerve terminals in the rat neostriatum. An ultrastructural study. AB - In the neostriatum 7.7% of all boutons absorbed alpha-methylnoradrenaline, thus staining their synaptic vesicles profiles granular. These presumably monoaminergic synapses were divided into two types according to ultrastructural features. Type A was composed of a relatively large axon terminal and a dendritic spine with postsynaptic membrane thickening. The axon terminal of type B, that is a bouton en passant, was smaller than the postsynaptic profile, while the synaptic contact was symmetrical. Following nigral coagulation, both types of synapse degenerated. It is concluded that both types of synapse are dopaminergic. PMID- 6270602 TI - Adrenaline neurons in the rostral ventrolateral medulla innervate thoracic spinal cord: a combined immunocytochemical and retrograde transport demonstration. AB - Adrenaline-containing neurons in the C1 group of the ventrolateral rostral medullary reticular formation which project to the thoracic spinal cord were identified by a combined retrograde transport immunocytochemical technique. No other medullary catecholamine neurons, including the A1 and A2 noradrenaline cells, project to thoracic spinal cord. These data, taken with results of other studies of spinal catecholamine innervation, suggest a segmental segregation of projections to spinal cord by dopaminergic, noradrenergic, and adrenergic neurons. PMID- 6270604 TI - Descending inhibitions from the nucleus raphe magnus and adjacent reticular formation to the dorsal horn of the rat are not antagonized by bicuculline or strychnine. AB - The effects of iontophoretically applied bicuculline and strychnine on brainstem and segmental-evoked inhibitions of dorsal horn cells have been examined in the rat. Both antagonists failed to reduce the descending inhibitory influences at concentrations which selectively blocked responses to GABA or glycine and segmentally-derived inhibitions of the same neurones. These results suggest that neither GABA nor glycine is involved in brainstem inhibition of rat dorsal horn cells. PMID- 6270603 TI - Separate populations of cholecystokinin and 5-hydroxytryptamine-containing neuronal cells in the rat dorsal raphe, and their contribution to the ascending raphe projections. AB - Successive immunohistofluorescent staining for cholecystokinin and 5 hydroxytryptamine (5-HT) was combined with fluorescent retrograde axonal tracing to analyze ascending projections from the rat dorsal raphe nucleus. Cholecystokinin-like immunoreactive cell bodies were situated both rostral and caudal to the 5-HT-immunoreactive perikarya in the dorsal raphe. Only the 5-HT immunoreactive neurones could be retrogradely labelled after injections of fluorescent axonal tracers into various forebrain areas. PMID- 6270605 TI - The cortical projections of the thalamic intralaminar nuclei, as studied in cat and rat with the multiple fluorescent retrograde tracing technique. AB - Two retrograde fluorescent tracers were injected in two different areas of the cerebral cortex in rats and in cats. In all the experiments many single labeled cells and only some double labeled ones were seen in the thalamic intralaminar nuclei. The present results suggest that the diffusely distributed intralaminar cortical projections mainly consist of axons of separate cells, and only to a minor extent of axon collaterals of the same cells. PMID- 6270607 TI - Presence of 1,25-dihydroxyvitamin D3 receptor in rat pituitary. PMID- 6270606 TI - Two types of synaptic facilitation recorded in pyramidal cells of in vitro hippocampal slices from guinea pigs. AB - PyramidaL cells in CA1 hippocampus show two types of increase in the excitatory postsynaptic potential (EPSP) during paired pulse stimulation of the afferent fibres in stratum radiatum. For 15-300 ms interstimulus intervals, both the initial slope and the peak amplitude of the EPSP are increased, which is mainly due to an increased excitatory synaptic current. For 300 ms-2 s intervals, the predominant change is an increase in the peak amplitude. The latter type of facilitation is caused by a decrease in a superimposed inhibitory postsynaptic potential (IPSP). PMID- 6270608 TI - [Effect of thyroxine on the activity of a series of mitochondrial enzymes during the process of development]. AB - The activity of alpha-glycerophosphate dehydrogenase and succinate dehydrogenase suffers almost no changes during the chick embryogenesis, whereas the activity of cytochrome oxidase increases by 20--25%. The activity of these enzymes in the embryos exceeded that in the adult birds. The injection of thyroxine during embryogenesis did not affect the activity of the enzymes under study but in chickens stimulated the activity of mitochondrial enzymes. The increase of activity of alpha-glycerophosphate dehydrogenase, cytochrome oxidase and succinate dehydrogenase was also noted in young rats. PMID- 6270609 TI - Fibrous histiocytomas of the oral mucosa. AB - The oral lesions reported in this article fall into the first category of benign fibrous histiocytomas analogous to those which occur on sun-exposed skin surfaces of young objects. The ages of the patients, the histologic features and the history of traumatic episodes, as well as their biologic behaviors on follow-up, are all features compatible with those benign lesions which occur in the skin. According to the information presented in these cases, there is stronger evidence that the lesions of the oral mucosa, like their dermatologic counterparts, are representative of reactive inflammatory processes rather than neoplastic processes. It is of considerable interest that in both cases reported here the patients were children whose lesions developed following significantly severe traumatic episodes. Also of interest is the finding that the lesions healed with no recurrences or complications, even though in one of the cases it was not completely removed. These findings are in agreement with other reports of such lesions which have occurred in the head and neck regions of children and young adults. From the over-all information obtained in the literature review regarding the biologic behavior of benign fibro-histiocytic lesions, the collected data seem to indicate that lesions of the skin and superfacial mucosal surfaces which occur in children and young adults are proliferative, reactive lesions and infection, or irradiation. Systemic, visceral, or deep-seated lesions in the lower extremities appear to be true neoplasms and their prognoses must be considered as guarded. PMID- 6270610 TI - Glomus tumor (golomangioma) of the tongue. A light and electron microscopic study. AB - A rare case of an intraoral glomus tumor on the ventral surface on the tongue was studied by light and electron microscopy. With light microscopy, the tumor was composed of dilated vascular channels surrounded by "epithelioid" glomus cells exhibiting large round nuclei and abundant eosinophilic cytoplasm. Bodian stains showed significant staining for nerve fibers among the tumor cells. Electron microscopic study demonstrated that the tumor cells had ultrastructural features which were similar to both smooth muscle cells and endothelial cells, suggesting that a "transitional" cell may give rise to glomus tumor. The literature on oral glomus tumors is briefly reviewed. PMID- 6270611 TI - [Reserve capacity of hypophyseal trophic hormones. Plasma ACTH levels and ACTH reserve in adrenal cortex diseases]. PMID- 6270612 TI - [Changes in angiotensin converting enzyme activity in bronchial asthma during physical exertion]. PMID- 6270613 TI - [Case of laryngeal myoblastoma]. PMID- 6270614 TI - Epstein-Barr virus infection and neoplasia. AB - Once acquired, Epstein-Barr virus (EBV), a latent virus, remains in the body for what appears to be the lifetime of the human host. Circumstantial data suggest EBV is involved in clinical disease including malignancies far more often than previously recognized. A serologic test for early antigen (EA) is more specific for diagnosing active EBV disease than the monospot or heterophile test. A case study of active Epstein-Barr infection is reported showing persistently elevated early antigen titers prior to and following malignant transformation. PMID- 6270615 TI - Prostaglandin E2, collagenase, and protease levels in culture of rat tumor. AB - The production of collagenase, protease, leucine aminopeptidase, and prostaglandin E2 by rat tumor increased with the growth of tumor. The syntheses of collagenase, protease, and leucine aminopeptidase were significantly inhibited by indomethacin. Inhibition of collagenase, protease, and leucine aminopeptidase syntheses was overcome by the addition of prostaglandin E2 at a concentration equivalent to the endogenous level of prostaglandin E2 in rat tumor. These findings suggested that prostaglandin E2, possibly derived from tumor cells, was involved in stimulating the tumor connective tissue stroma to produce collagenase, protease, and leucine aminopeptidase, which promoted tumor cell invasion. PMID- 6270616 TI - Recurrent benign mixed tumor of lacrimal gland: report of a case with intracranial extension. AB - A 66-year-old man with benign mixed tumor of the lacrimal gland underwent an excisional biopsy in 1966. Three recurrences subsequently developed, which resulted in an orbital exenteration. In 1979 he came to us. We discovered radiographic evidence and surgical confirmation of recurrent disease involving the lesser wing of the sphenoid and dura in the floor of the anterior cranial fossa. The natural history and management of this tumor is discussed and the relevant literature is reviewed. PMID- 6270617 TI - Angiofibroma: a rare case in an elderly female. PMID- 6270618 TI - [Basic principles of hypoxia control during shock]. PMID- 6270619 TI - [Presence of intracellular collagen fibrils in systemic lymph node neoplasms]. PMID- 6270620 TI - [Structural and ultrastructural findings concerning pancreatic neoplasms associated with the WDHA syndrome (watery diarrhea-hypokalemia-achlorhydria syndrome)]. PMID- 6270621 TI - [Case contribution to our knowledge of syncytial endomyometritis]. PMID- 6270622 TI - [Malignant mesenchymal tumors of the larynx]. PMID- 6270623 TI - Release of vasopressin by the fetal lamb during premature parturition induced with corticotropin. AB - These studies were implemented to assess the relationship between fetal vasopressin secretion and the progression of parturition as well as the contribution of specific stimuli to vasopressin release during labor. In chronically catheterized fetal lamb preparations, labor was induced by infusion of adrenocorticotropin (12.5 mg/kg/hr) to seven fetuses at 130 +/- 1 day of gestation. Before labor, fetal plasma vasopressin concentrations were 2.1 +/- 1.4 pg/ml and remained low (5.3 +/- 3.4 pg/ml) during prodromal and early phases of labor, but rose significantly in the active and expulsive phases (39.6 +/- 27.5 and 173.3 +/- 152.9 pg/ml) to reach peak values at delivery (584.2 +/- 433 pg/ml) and decrease by 30 min after birth (359.8 +/- 90.0 pg/ml). At delivery, fetal plasma vasopressin concentrations were strongly correlated (P less than 0.001) with hormone values obtained during the latter phases of labor. Fetal arterial pH and oxygen tension was inversely correlated with plasma vasopressin (P less than 0.01). No similar correlations were found with arterial PCO2, K, Na, Cl, osmolality, or packed cell volume. Unexpectedly, we observed a significant (P less than 0.001) and progressive decrease in fetal oxygen tension during the induction process. Other characteristics of adrenocorticotropin-induced parturition seemed to mimic those of spontaneous labor. PMID- 6270624 TI - [Current theories on the etiology of Crohn disease]. PMID- 6270625 TI - The immune system in slow disease due to persistent submicrobial (viral) infection. PMID- 6270626 TI - Facilitation of acetylcholine release from cardiac parasympathetic nerve endings. Effect of stimulation pattern and Mn ions. AB - The influence of the stimulus intervals and the effect of Mn ions on facilitation of acetylcholine (ACh) release from parasympathetic nerve terminals were studied in quiescent guinea-pig auricles by electrophysiological methods. A maximum facilitation occurs at intervals of about 50ms. The half time of decay of facilitation after a conditioning stimulus is about 500ms. When conditioning trains of stimuli were applied, a second much longer lasting component of facilitation was found (t1/2 congruent to 4s). Mn ions, after exerting an inhibitory effect, cause an increase of ACh release, the development of which is dependent on frequent stimulation of the nerve fibres. This potentiation is accompanied by an apparent loss of facilitation. A further increase in ACh release occurs when superfusion is changed from Mn containing to normal Tyrode's solution. The decay to the control level displays a half time of about 20 min and can also be accelerated by frequent stimulation of the parasympathetic nerve fibres. It is suggested that Mn ions not only inhibit a Ca inward current but may also act on intracellular Ca2+ bindings sites in the nerve terminal. When these sites are blocked even a reduced Ca influx can be more effective in the process of transmitter release. PMID- 6270627 TI - Absorption of Na, H ions and short chain fatty acids from the sheep colon. AB - The interrelationship between the absorption of Na, H ions and short chain fatty acids (SCFA) and the secretion of HCO3 in the sheep colon was studied using an in vivo perfusion technique. 1. Under control conditions net SCFA absorption was 62.9 +/- 6.1 mmol . h-1, net Na absorption 49.8 +/- 7.6 mmol . h-1. HCO3 accumulated in the colon lumen at a rate of 5.1 +/- 2.9 mmol . h-1. 2. When 30, 50, or 70% of the Na in the perfusion solution were replaced by Li, Na transport was gradually inhibited and even abolished when 70 mmol . 1(-1) of Li were present. SCFA absorption also decreased, however, to a lower degree. 3. Net Na transport was higher in the presence than in the absence of SCFA. This difference disappeared when Na transport was inhibited by Li. 4. HCO3 accumulation in the colon lumen significantly increased with increasing Li concentrations up to a maximum of 21.7 +/- 7.1 mmol . h-1 when 50% of Na were replaced by Li, however, only in the presence of SCFA. 5. Neither increasing Li concentrations nor the presence or absence of SCFA influenced the H ion disappeared from the perfusion solution. 6. These results indicate that net absorption of Na and SCFA in sheep colon are closely linked to the availability of H ions. PMID- 6270628 TI - Catecholamine-induced vasodilator metabolite release from guinea-pig hearts is not due to increased myocardial activity. AB - Guinea-pig isolated hearts were perfused in series, the donor heart perfusate supplying the recipient heart. Isoprenaline increased the rate and force of contraction and oxygen consumption of donor hearts and produced a coronary vasodilatation. This was accompanied by the release of vasodilator metabolite as demonstrated by vasodilatation of the recipient heart, the beta-adrenoceptors of which were antagonized by propranolol. During arrest of donor hearts by either carbachol or application of a fibrillating current, isoprenaline still released vasodilator metabolite and increased oxygen consumption but without changes in rate or tension. This release was prevented by beta-adrenoceptor blockade. It is concluded that the sympathomimetic-induced coronary vasodilatation is mediated via the release of a vasoactive metabolite, the trigger for which is not the concomitant mechanical hyperactivity. PMID- 6270629 TI - Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches. AB - 1. The extracellular patch clamp method, which first allowed the detection of single channel currents in biological membranes, has been further refined to enable higher current resolution, direct membrane patch potential control, and physical isolation of membrane patches. 2. A description of a convenient method for the fabrication of patch recording pipettes is given together with procedures followed to achieve giga-seals i.e. pipette-membrane seals with resistances of 10(9) - 10(11) omega. 3. The basic patch clamp recording circuit, and designs for improved frequency response are described along with the present limitations in recording the currents from single channels. 4. Procedures for preparation and recording from three representative cell types are given. Some properties of single acetylcholine-activated channels in muscle membrane are described to illustrate the improved current and time resolution achieved with giga-seals. 5. A description is given of the various ways that patches of membrane can be physically isolated from cells. This isolation enables the recording of single channel currents with well-defined solutions on both sides of the membrane. Two types of isolated cell-free patch configurations can be formed: an inside-out patch with its cytoplasmic membrane face exposed to the bath solution, and an outside-out patch with its extracellular membrane face exposed to the bath solution. 6. The application of the method for the recording of ionic currents and internal dialysis of small cells is considered. Single channel resolution can be achieved when recording from whole cells, if the cell diameter is small (less than 20 micrometer). 7. The wide range of cell types amenable to giga-seal formation is discussed. PMID- 6270630 TI - [Computed tomographic angiography (CT angio) of hepatic tumor: experimental and clinical studies (author's transl)]. PMID- 6270631 TI - [Neuropathy induced by amiodarone]. PMID- 6270632 TI - [Pneumocystis carinii pneumonia in patients with lung neoplasms]. PMID- 6270633 TI - Purification and characterization of human converting enzyme (kininase II). AB - We purified peptidyl-dipeptidase (converting enzyme, EC 3.4.15.1) to homogeneity from the membrane fraction of human lung and for comparison, from human and hog kidney. The membrane-bound lung enzyme was purified 1800-fold with 19% yield, and the kidney enzyme 640-fold with 10% yield. The specific activities with Bz-Gly His-Leu were 81 mumol/min/mg for the lung and 65 for the kidney enzyme. The lung enzyme was homogeneous in gel electrophoresis with Mr = 155,000 and Sw,20 = 8.0 in ultracentrifugation. Antibodies elicited against lung or kidney enzyme cross reacted with enzyme from other organ, but not with the hog enzyme. In isoelectric focusing both human enzymes had a major form with a pI of 5.2. The lung preparation also contained more acidic forms (pI = 4--5), which were eliminated by treatment with neuraminidase. Lung and kidney converting enzyme hydrolyzed bradykinin, angiotensin I, and enkephalins and had similar kinetic constants. Bradykinin was the best substrate, as indicated by its kcat/Km, but Met5 enkephalin had the highest turnover number. The hydrolysis of Bz-Gly-His-Leu was inhibited by captopril (SQ 14225) competitively, and by Keto-ACE, a non-peptide derivative of Bz-Phe-Gly-Pro, non-competitively. PMID- 6270634 TI - Hypothermic and antipyretic effects of centrally administered ACTH (1--24) and alpha-melanotropin. AB - ACTH (1--24) and alpha-melanotropin (alpha-MSH), peptides previously shown to influence body temperature when administered centrally and to occur naturally in brain regions important to temperature control, were injected intracerebroventricularly (ICV) in rabbits. The peptides in doses of 1.25, 2.5 and 5.0 micrograms produced dose-related hypothermias in a 23 degrees C environment, and greater decreases in body temperature when the experiments were repeated in the cold (10 degrees C), but the largest dose had no effect on temperature in the heat (30 degrees C). These results indicate that the peptides do not reduce the central set-point of temperature control. Rather, they appear to selectively inhibit heat conservation and production responses. Five microgram of ACTH reversed vasoconstriction and inhibited rises in temperature caused by leukocytic pyrogen (LP) given IV and ICV. The same dose of alpha-MSH also reduced fever produced by IV and ICV LP, but the reduction was not as great as after ACTH. Both peptides (5 micrograms) also reduced temperature rises and vasoconstriction caused by ICV PGE2. ACTH reduced d-amphetamine-induced hyperthermia without altering vasoconstriction which suggests that this peptide can reduce temperature rises by inhibiting heat production alone. One of the most important findings was that the peptides are antipyretic in that they reduce fever at doses (0.25 microgram, ICV) that do not affect normal temperature. The powerful effects of these peptides on resting body temperature, hyperthermia and fever, together with their presence in brain tissue important to temperature control, suggest that the endogenous central peptides participate in thermoregulation, perhaps by limiting fever and influencing normal temperature. PMID- 6270635 TI - Neuromuscular response of the immature rat of ACTH/MSH 4--10. AB - The adrenocorticotropin fragment ACTH/MSH 4--10 (0.1 ug/kg IP) effectively modulates the neuromuscular responses of 9 to 15 day old rats. Muscle (extensor digitorum longus) contraction amplitude is increased, fatigue is delayed and muscle half-relaxation time is shortened during 20 min of continuous in situ stimulation of a branch of the deep peroneal nerve (square wave shocks 10 Hz, duration 0.5 msec, strength supermaximal). No effect on contraction time is seen. There is no facilitation or change in any contraction parameter in rats older than two weeks (16 to 40 days) indicating that these older animals, like normal adult rats, are unaffected by the peptide. Immature rats, however, are even more sensitive than hypophysectomized adult rats [29] to the ameliorative action of ACTH/MSH 4-10. This early sensitivity to ACTH/MSH 4--10 corresponds to important developmental changes occurring in nerve and muscle during the most critical period in postnatal development, the first two weeks. PMID- 6270636 TI - Action of selected serotonin antagonists on hyperthermia evoked by intracerebrally injected beta-endorphin. AB - Methergoline, an antagonist of cerebral serotonin receptors, has been shown to significantly reduce the rise in rectal temperature (Tre) produced by the intracerebral microinjection of beta-endorphin. In this study the role of serotonin in the increase in Tre elicited by beta-endorphin was further examined using three additional serotonin antagonists. beta-Endorphin was administered twice to rats using a crossover design in which half of the animals were first pretreated with the vehicle solution and half with the antagonist. Serotonin antagonists used were: methergoline, methysergide, cinanserin and cyproheptadine. Although methergoline did cause a marked reduction in the beta-endorphin-induced rise in Tre, neither methysergide, nor cinanserin, nor cyproheptadine produced a marked reduction in the hyperthermia. Since methergoline also interacts with the dopamine receptor, the effect of a dopamine antagonist, haloperidol, on the endorphin-evoked response was also examined. Haloperidol failed to attenuate the rise in Tre. The reason for the apparent discrepancy in the action of these serotonin antagonists is unclear. Further research may reveal distinct subpopulations of serotonin receptors at which these antagonists exert differential effects. PMID- 6270637 TI - [Pneumotropic viruses in chronic respiratory tract diseases in children]. PMID- 6270638 TI - [Preliminary evaluation of cisplatin in the treatment of malignant testicular neoplasms other than seminomas]. PMID- 6270639 TI - [Cytochemical studies of peripheral blood granulocytes after occupational exposure to pesticides]. PMID- 6270640 TI - [Pathogenesis and diagnosis of idiopathic hypercalciuria]. PMID- 6270641 TI - [Regulation of cyclic AMP and cyclic GMP-dependent protein kinase activity]. PMID- 6270642 TI - [Proteolytic enzymes of the blood platelets]. PMID- 6270643 TI - Insulinoma masquerading as carotid transient ischaemic attacks. AB - A patient who initially presented with classical internal carotid territory transient ischaemic attacks, on further investigations was found to have an insulinoma, the removal of which completely alleviated the symptoms. This appears to be an unusual form of presentation of a rare tumour. The possible mechanisms of focal neurological disturbances during hypoglycaemia are discussed. PMID- 6270644 TI - [Distribution of hypothalamic cholinergic and adrenergic system controlling hypophyseal-adrenal system function]. AB - It has been shown on rat experimental models that the cholinergic systems, participating in hypothalamic regulation of the hypophyseal adrenocortical function, are localized in the posterior hypothalamus whereas alpha-adrenergic systems in the posterior and anterior hypothalamus, and beta-adrenergic systems in the ventromedial hypothalamus. PMID- 6270645 TI - [Effect of the adrenocorticotropic hormone on peripheral blood levels of hydrocortisone, testosterone and their precursors in baboons]. AB - The time-course of changes in hydrocortisone, 17-hydroxypregnenolone, 17 hydroxyprogesterone, dehydroepiandrosterone and testosterone levels in the peripheral blood of male sacred baboons was studied after exogenous ACTH injection by radioimmunoassay. It was shown that the hydrocortisone secretion character following ACTH injection correlates with the change of other adrenal steroids studied: 17-hydroxypregnenolone, 17-hydroxyprogesterone and dehydroepiandrosterone. Maximum concentration growth of the above steroids is seen 2 hours after ACTH injection and is 90, 80, 210 and 60%, correspondingly. Dynamics of testosterone response does not correlate with the adrenal steroid concentration change. Testosterone content is increased by 160% 30 minutes after ACTH injection, reaching 60% of the initial level following 6 hours. PMID- 6270646 TI - [Role of viruses in the etiology of endocrine diseases]. PMID- 6270647 TI - [Hormonal regulation of vitamin D transformation into active metabolites]. PMID- 6270648 TI - [Function of the neuromotor apparatus in diabetes mellitus depending on metabolic compensation]. AB - Electromyography was used to show that metabolism decompensation in diabetes mellitus induces reversible muscular changes, manifesting in the decreased tendon reflex and motor response amplitudes. These indices of neuromotor apparatus function return to normal in diabetes compensation. Meanwhile, diabetes compensation does not lead to rapid recovery of the affected nervous conduction and spinal motoneuronal excitability. PMID- 6270649 TI - [Various adrenergic reactions in experimental hyperthyroidism]. AB - Three adrenergic responses, i. e. noradrenaline calorigenic effect, positive isoproterenol chronotropic action and seminiferous duct contraction after noradrenaline use was studied in experimental albino male rats. It was shown that hyperthyroidism provokes intensification of the beta-adrenergic responses examined (noradrenaline calorigenic effect and positive isoproterenol chronotropic effect); beta-adrenoreceptor sensitivity remained unchanged. Experimental hyperthyroidism was accompanied by a decrease in alpha1 adrenoreceptor sensitivity without changing the intensity of this adrenergic response. PMID- 6270650 TI - [Functional activity of mitochondria in experimental hypoparathyroidism]. AB - Respiration and oxidative phosphorylation of the brain and hepatic mitochondria, as well as the activity of succinate dehydrogenase, succinate-cytochrome-c reductase and cytochrome oxydase were studied in experimental hypoparathyrosis. Activated respiration and decreased effectiveness of the brain and hepatic mitochondrial phosphorylation and enhanced succinate dehydrogenase activity are seen during hypoparathyrosis development. It is suggested that alterations in mitochondrial functional activity are caused not only by the changed blood calcium ion number, but also by hypoxia, developing in experimental hypoparathyrosis. PMID- 6270651 TI - [Regulation of aldosterone secretion]. PMID- 6270652 TI - Mechanism for transcriptional action of cyclic AMP in Escherichia coli: entry into DNA to disrupt DNA secondary structure. AB - Binding analysis with purified bacterial receptor distinguishes two structural domains in cyclic AMP (cAMP). The first, the cyclic phosphate and furanose, constitutes a binding domain. This region is bound tightly to the receptor. The rest of cAMP is not bound; the adenine moiety of cAMP is exposed. Unlike binding, activity of cAMP requires the adenine moiety. To be active, cAMP must have in domain II the base adenine--specifically, its Watson--Crick atoms N-1 and N-6. Analysis of indoleacetic acid, a compound able to replace cAMP at the L-arabinose operon, indicates a similar distinction between binding and active domains. To be active, the indole must have substitution (carboxyl or amide) electronically comparable to the cAMP N-1 and N-6. On this basis, we propose a detailed mechanism for action of cAMP (or indoleacetic acid) in Escherichia coli. We propose that the exposed adenine of cAMP enters into the DNA. The adenine's N-1 and N-6 form hydrogen bonds to a thymine in DNA. This interaction destabilizes the DNA. It enhances transcription. Marked similarities indicate an identical mechanism for the steroid hormones in eukaryotes. PMID- 6270653 TI - Discrete-length repeated sequences in eukaryotic genomes. AB - Two of the four repeated DNA sequences near the 5' end of the silk fibroin gene hybridize with discrete-length families of repeated DNA. These two families comprise 0.5% of the animal's genome. A repeated sequence with a conserved length has also been found in the short class of moderately repeated sequences in the sea urchin. The discrete length, interspersion, and sequence fidelity of these moderately repeated sequences suggests that each has been multiplied as a discrete unit. Thus, transposition mechanisms may be responsible for the multiplication and dispersion of a large class of repeated sequences in phylogenetically diverse eukaryotic genomes. The repeat we have studied in most detail differs from previously described eukaryotic transposable elements: it is much shorter (1300 base pairs) and does not have terminal repetitions detectable by DNA hybridization. A simple technique for identifying such discrete-length repeated sequences is described. PMID- 6270654 TI - Solubilization of human platelet alpha-adrenergic receptors: evidence that agonist occupancy of the receptor stabilizes receptor--effector interactions. AB - The alpha-adrenergic receptors of human platelet membranes can be directly identified by both a radiolabeled agonist, [3H]epinephrine, and a radiolabeled antagonist, [3H]yohimbine. Digitonin solubilizes a binding component from the membrane that is indistinguishable from the alpha-receptor identified in the native platelet membrane as assessed by (i) order of potency of agonists and antagonists and (ii) affinity of the receptor for [3H]-yohimbine and competing antagonists. However, the solubilized receptor demonstrates a reduced affinity for agonists and a loss of the ability of guanine nucleotides to modulate receptor affinity for agonists. Prelabeling of human platelet membranes with [3H] epinephrine results in a guanine nucleotide-sensitive agonist-receptor complex that sediments more rapidly in sucrose gradients than do unoccupied or antagonist occupied receptors. Thus, agonist occupancy of the alpha-receptor prior to membrane solubilization may promote or stabilize receptor interaction with effector components in the membrane, one of which may be the GTP regulatory protein responsible for modulation of receptor affinity. PMID- 6270655 TI - Recombinant bacteriophages containing the integrated transforming provirus of Gardner--Arnstein feline sarcoma virus. AB - The integrated DNA provirus of the Gardner-Arnstein (GA) strain of feline sarcoma virus (FeSV) was molecularly cloned in a bacteriophage lambda vector. The cloned DNA fragment is 14.4 kilobase pairs long and contains a 6.7-kilobase provirus flanked by cellular sequences derived from nonproductively transformed mink cells. Transfection of mouse NIH/3T3 cells with the cloned DNA fragment induced foci of transformation at efficiencies of 10(4) focus-forming units/pmol of sarcoma virus DNA. Restriction endonuclease mapping and heteroduplex analyses were used to compare the GA-FeSV provirus with that of Snyder-Theilen (ST)-FeSV, a second strain that contains homologous transformation-specific sequences (v fes). Both viruses have the general structure 5'-gag-fes-env-c region-3', each having retained portions of the feline leukemia virus (FeLV) gag and env genes. In addition to segments shared by the two sarcoma viruses, GA-FeSV contains 1.7 kilobases of extra sequences not found in ST-FeSV. Of these, at least 400-500 base pairs located near the 5' end of v-fes encode a portion of the GA-FeSV polyprotein; the remaining 1.2 kilobases are derived from the FeLV env gene but do not appear to encode any detectable product related to the FeLV envelope glycoprotein. The close homology of the v-fes sequences shows that GA- and ST FeSV were formed by recombination of FeLV with similar portions of a cat cellular gene (c-fes). PMID- 6270656 TI - Transposon Tn3 encodes a site-specific recombination system: identification of essential sequences, genes, and actual site of recombination. AB - The bacterial transposon Tn3 encodes a site-specific recombination system. The recombination requires the product of tnpR, a gene previously identified as a repressor of the transposase. This recombination is site specific and takes place somewhere within the sequence C-G-A-A-A-T-A-T-T-A-T-A-A-A-T-T-A-T-C but requires at least one additional sequence outside this. The phenotype of mutations in this recombination system suggests that transposition proceeds by a mechanism in which cointegrates are intermediates. PMID- 6270657 TI - Energy-dependent reversal of the cytochrome oxidase reaction. AB - Energization of isolated rat liver mitochondria with ATP under conditions in which cytochrome c is poised in a highly oxidized state shifts the state of cytochrome oxidase (cytochrome c oxidase; ferrocytochrome c:oxygen oxidoreductase, EC 1.9.3.1) from fully oxidized to two new spectroscopically distinguishable states depending on the applied phosphorylation potential and redox potential at cytochrome c. Both new states are spectrally similar or identical to two previously described intermediates in the reaction between reduced enzyme and O2. The data suggest that the energy-dependent transitions are due to reversed electron transfer from water to ferricytochrome c linked to accumulation of intermediates of O2 reduction at the catalytic heme a3/copper center. Titrations with redox potential indicate that each transition is a one electron step, a finding that would identify the second observed compound as enzyme-bound peroxide or its equivalent. This is consistent with this compound being spectrally identical to "Compound C," previously described as the reaction product between half-reduced oxidase (two electrons) and O2. On the basis of these data a catalytic scheme of O2 reduction is proposed for the heme a3/copper center of cytochrome oxidase. PMID- 6270658 TI - New segment-coupling method for peptide synthesis in aqueous solution: application to synthesis of human [Gly17]-beta-endorphin. AB - The thiolcarboxyl peptide [17-thiolglycine]-beta-endorphin-(1-17) (I) was synthesized by the solid-phase method. Reaction of peptide I with citraconic anhydride gave citraconyl-[Lys(Cit9,GlyS17]-beta-endorphin-(1-17) (Ia). Peptide Ia was coupled to another synthetic peptide, [Lys(Cit)19,24,28,29]-beta-endorphin (18-31), by reaction with silver nitrate--N-hydroxysuccinimide in water. All citraconyl groups were removed in aqueous acetic acid, and [Gly17]-beta-endorphin was isolated in 30-40% yield. The synthetic analog had 10% analgesic potency and 59% opiate--receptor binding activity when compared with human beta-endorphin. PMID- 6270659 TI - Phosphorylation of ribosomal protein S6 in avian sarcoma virus-transformed chicken embryo fibroblasts. AB - Protein phosphorylation was examined in whole cell extracts from normal and avian sarcoma virus-transformed chicken embryo fibroblasts. The addition of serum or epidermal growth factor to serum-starved normal cells resulted in increased 32P labeling of a Mr 30,000 protein. In extracts from cells transformed by a temperature-sensitive mutant of Schmidt-Ruppin virus, subgroup A, and grown at the permissive temperature, the protein was phosphorylated regardless of serum starvation. This Mr 30,000 protein was shown to be ribosomal protein S6, and the effects of avian sarcoma virus transformation on S6 phosphorylation were further investigated. The ability to phosphorylate S6 in the absence of serum was found to be temperature sensitive when S6 preparations from the temperature-sensitive mutant-infected cells incubated at permissive and nonpermissive temperatures were compared. Cells transformed by the parent virus (Schmidt-Ruppin, subgroup A) maintained the ability to phosphorylate S6 in the absence of serum when incubated at either temperature. Phosphoserine was the only phospho-amino acid detected in acid hydrolysates from phosphorylated S6 preparations. PMID- 6270660 TI - Novel opiate binding sites selective for benzomorphan drugs. AB - The simultaneous addition of [D-Ala2, D-Leu5]-enkephalin and morphiceptin at concentrations at which 98% of enkephalin (delta) and morphine (mu) receptors are occupied only partially inhibits the binding of [3H]diprenorphine to rat brain membranes. These conditions, furthermore, do not affect the curves for displacement of [3H]diprenorphine binding by unlabeled diprenorphine. These data suggest that [3H]diprenorphine binds to a third subtype of opiate binding site, which has high affinity for diprenorphine but very low affinity for mu and delta agonists. The [3H]-diprenorphine binding observed in the presence of morphiceptin and [D-Ala2, D-Leu5]enkephalin exhibits high affinity for several benzomorphan drugs in the chemical family of 6,7-benzomorphan (e.g., cyclazocine, ethylketocyclazocine, SKF 10047, UM 1072, oxilorphan, etc). Because of its selectivity for most benzomorphan drugs, this putative receptor site is tentatively referred to as a benzomorphan binding site. Its regional distribution in rat brain is similar to that of morphine (mu) receptors but differs from that for enkephalin (delta) receptors. The content of benzomorphan binding sites in rat brain is only one-half to one-third that of morphine receptors. The relative affinities of various opioids to morphine enkephalin, and benzomorphan binding sites are also described. PMID- 6270661 TI - Site-specific interaction of DNA gyrase with DNA. AB - DNA gyrase, in the presence of the inhibitor oxolinic acid, can induce double strand DNA breakage at specific sites. The sequences at several sites have been determined. In addition, the structure of complexes formed between DNA gyrase and restriction fragments containing an oxolinic acid-promoted cleavage site has been examined by DNase protection methods. DNA gyrase protects more than 120 base pairs of DNA against pancreatic DNase in a region surrounding the cleavage site. Protection is observed both in the presence and absence of oxolinic acid. Protected DNA flanking the cleavage site contains DNase I-sensitive sites spaced on the average 10 or 11 base pairs apart. This result supports the view that, in the DNA gyrase--DNA complex, the DNA is largely wrapped on the outside of the enzyme. PMID- 6270662 TI - Molecular cloning and partial characterization of unintegrated linear DNA from gibbon ape leukemia virus. AB - We have cloned the complete genome of an oncogenic primate retrovirus, the San Francisco isolate of gibbon ape leukemia virus, in a lambda phage vector. DNA sequence analysis and restriction endonuclease mapping of the inserted linear provirus demonstrated 9-base pair inverted repeats at its ends, flanking direct terminal repeats 470 base pairs in length. The (-) strong stop region of this DNA showed surprisingly low sequence homology to that of another gibbon ape leukemia virus isolate from an animal with similar disease. Analysis of the clone also revealed the terminal phosphate configuration of the linear provirus. The recombinant phage is suitable for direct use as a hybridization probe to detect homologous retroviral sequences in human cell lines. PMID- 6270664 TI - Radioimmunochemical measurement of the transferrin receptor in human trophoblast and reticulocyte membranes with a specific anti-receptor antibody. AB - A radioimmunoassay was developed to directly assay the presence of transferrin receptors in human tissues. Antisera developed in a goat against purified human placental transferrin binding protein was purified by fractional sodium sulfate precipitation and adsorption against Sepharose-bound transferrin to remove trace anti-transferrin activity. The antisera immunoprecipitates a Mr 94,000 peptide on 125I-iodinated syncytial trophoblast membranes from placentae. This polypeptide has been identified previously as the transferrin binding protein of the placenta [Wada, H. G., Hass, P. E. & Sussman, H. H. (1979) J. Biol. Chem. 254, 12629 12635]. A standard curve using purified 125I-iodinated placental transferrin receptor and various amounts of the purified noniodinated receptor is sensitive from 5 to 900 ng. A reticulocyte-enriched membrane ghost preparation (5% reticulocyte) gives a value of 9.5 micrograms of receptor per mg of protein. Normal erythrocyte membrane ghosts show binding (0.57 micrograms of receptor per mg of protein) proportional to the amount of reticulocytes normally present in blood (0.5-1.0%). In other tissues in which the transferrin receptor binding has been reported, purified syncytial trophoblastic membranes are found to have 34.5 micrograms of receptor per mg of protein, and BeWo cells, a choriocarcinoma cell line, are found to have 15.7 micrograms of receptor per mg of protein. In contrast, normal breast tissue, which has no demonstrated transferrin binding, contains only 0.18 micrograms of receptor per mg of protein by this method. PMID- 6270663 TI - A nucleotide change at a splice junction in the human beta-globin gene is associated with beta 0-thalassemia. AB - beta 0-Thalassemia is a heterogeneous group of disorders associated with absence of beta-globin. In a survey of DNAs from patients with beta 0-thalassemia of diverse ethnic origins, a change at the splice junction at the 5' end of the large intervening sequence (IVS 2) of the human beta-globin gene has been found in one patient of Italian and another two of Iranian ethnic origins. The enzyme Hph I recognizes a change at this site and generates a large-than-normal fragment of DNA, which hybridizes specifically to a beta-globin IVS 2 probe. No other changes in beta-globin gene DNA structure or organization are detectable by extensive restriction endonuclease analysis. The enzyme HinfI which recognizes a sequence beginning three nucleotides from the 5' end of the IVS 2 splice junction, produces normal fragments and localizes the defect to a G-G-T sequence at the 5'-end IVS 2 splice junction. This sequence is known to be remarkably conserved in all globin genes from many species and in most other genes examined to date. Thus, in at least some beta 0-thalassemia patients, the beta 0 thalassemia defect is associated with a nucleotide change at a splice junction. These patients provide unique examples of naturally occurring defects in splice junctions of eukaryotic genes associated with absence of specific gene function. PMID- 6270665 TI - Rapid purification of a high-affinity plasminogen activator from human blood plasma by specific adsorption on fibrin/Celite. AB - A preparation of fibrin precipitated over a solid Celite (diatomaceous earth) matrix that selectively binds 50-70% of the plasminogen activator present in human blood plasma is described. Affinity chromatography of plasma on fibrin/Celite followed by gel filtration led to a 29,000-fold purification of the plasminogen activator. The activator, referred to as the high-affinity plasminogen activator, is characterized by its ability to be strongly adsorbed by fibrin. Smaller amounts of other plasminogen activators and essentially all plasminogen were not bound to fibrin. The high-affinity plasminogen activator is a single-chain unstable protease with a molecular weight of 65,000-70,000. The high-affinity plasminogen activator has a low specific activity (500 CTA units/mg) compared to tissue or urine plasminogen activators (100,000-200,000 CTA units/mg) (CTA, Committee on Thrombolytic Agents). PMID- 6270666 TI - Sequences of the ssb gene and protein. AB - We have determined the sequences of the ssb gene and protein of Escherichia coli. The coding region of ssb is 534 base pairs and is preceeded and followed by dyad symmetries of 39 base pairs and 27 base pairs, respectively. The promoter for ssb is close to that for uvrA and these two genes are transcribed in opposite directions: ssb clockwise and uvrA counterclockwise on the standard E. coli genetic map. The DNA helix-destabilizing protein encoded by the ssb gene (single strand binding protein) contains 177 amino acids and has a calculated molecular weight of 18,873. Although there is no extensive sequence homology among the three helix-destabilizing proteins whose sequences are now known, both the E. coli and bacteriophage T4 DNA helix-destabilizing proteins do contain an acidic, alpha-helical region at their carboxy termini that may be functionally homologous. The remainder of the E. coli helix-destabilizing protein can be divided into two apparent domains on the basis of its amino acid sequence. The amino-terminal region (residues 1-105) contains 79% of the charged residues (27 out of 34 total) in the protein and is predicted to have a high degree of secondary structure (alpha helix and beta pleated sheet). In contrast, the region including residues 106-165 contains only two charged amino acids and is devoid of alpha helix or beta pleated sheet. PMID- 6270667 TI - Somatomedin receptor of human placenta: solubilization, photolabeling, partial purification, and comparison with insulin receptor. AB - Using a recently isolated human basic somatomedin (basic SM) similar to insulin like growth factor I (IGF-I), we studied both the photoaffinity-labeled and unlabeled basic-SM receptor solubilized from human placental cell membranes. Unlike the result with the insulin receptor, high yields of soluble basic-SM binding activity are obtained with Triton X-100. The soluble basic-SM receptor retains high-affinity (Kd approximately 0.3 nM) peptide-specific binding of basic SM, similar to the binding present in particulate placenta membranes; the receptor exhibits a comparatively low affinity for insulin (Kd approximately 3 microM). On Sepharose 6B, like the crude soluble insulin receptor, the basic-SM receptor migrates as a species with an apparent Stokes radius of 7.2 nm; unlike the insulin receptor, the basic-SM receptor does not, under similar conditions, yield a smaller binding species (apparent Stokes radius 3.8 nm). Upon photoaffinity labeling with 125I-labeled basic SM, one principal specifically labeled constituent is detected. Upon gel electrophoresis in the presence of 2 mercaptoethanol, the photolabeled constituent, like the insulin receptor, migrates as a species with an apparent molecular weight of about 140,000; in the absence of reducing agent, a molecular weight greater than 240,000 is observed. Lectin-agarose affinity chromatography yields a 30-fold purification both of the basic-SM-binding activity and the photolabeled constituent. Anti-insulin receptor antibody does not appear to precipitate the basic-SM receptor. We conclude that the basic-SM receptor of human placenta is a glycoprotein, remarkably similar to (an isoreceptor) but distinct from the insulin receptor previously characterized in this tissue. PMID- 6270668 TI - Characterization of a membrane-associated receptor from bovine liver that binds phosphomannosyl residues of bovine testicular beta-galactosidase. AB - A receptor that binds the phosphomannosyl recognition marker of bovine testicular beta-galactosidase (beta-D-galactoside galactohydrolase, EC 3.2.1.23) was isolated from bovine liver membranes. The receptor was extracted from crude plasma membrane preparations with Triton X-100 and immunoprecipitated as a receptor--beta-galactosidase complex with anti-beta-galactosidase. The receptor was dissociated from the precipitate with mannose 6-phosphate, labeled with 125I, and purified on a beta-galactosidase-Sepharose 4B affinity matrix. A quantitative binding assay employing anti-beta-galactosidase and IgGsorb (formalin-fixed Staphylococcus aureus) was devised to study the binding of 125I-labeled receptor to beta-galactosidase. Maximal binding of receptor to enzyme occurred at pH values between 5.7 and 6.5. Divalent cations were not required for binding. The values of the dissociation constant obtained for beta-galactosidase varied between 200 nM observed with "lower uptake" forms and 20 nM for "higher uptake" forms of the enzyme. A number of phosphorylated monosaccharides were tested as inhibitors of binding of enzyme to receptor; mannose 6-phosphate and fructose 1 phosphate served as inhibitors and exhibited Ki values of 0.064 mM and 0.24 mM, respectively. The receptor has a subunit molecular weight of 215,000. Similar receptors were also demonstrated in Triton X-100 extracts of human skin fibroblasts, Chinese hamster ovary cells, and rat hepatocytes. These cell types are known to assimilate lysosomal enzymes containing covalently bound mannose 6 phosphate residues. PMID- 6270670 TI - Reciprocity or near-reciprocity of highly coupled enzymatic processes at the multidimensional inflection point. AB - It has recently been demonstrated that coupled enzymatic processes may possess, for a particular choice of the state variables, multidimensional inflection points in thermodynamic force-flow space. The conditions for reciprocity in the linear region near such a reference state, which may be far from equilibrium, are of considerable interest. It is shown by examining the associated Hill diagrams that all cycles in which a given pair of forces act contribute a corresponding pair of symmetrical terms to the Jacobian matrix characterizing perturbations about this stationary state. To the extent that these cycles dominate--i.e., to the extent that the system is highly coupled--reciprocity or near-reciprocity will be obeyed. This would be expected to be the case in most biological systems. PMID- 6270669 TI - A Rous sarcoma virus provirus is flanked by short direct repeats of a cellular DNA sequence present in only one copy prior to integration. AB - The Rous sarcoma virus (RSV)-transformed rat cell line RSV-NRK-2 contains a single complete RSV provirus. We have obtained recombinant lambda clones that contain both ends of the RSV provirus and the flanking rat sequences. The provirus is integrated in unique DNA and is present in only one of the two homologous chromosomes. The rat sequences into which the RSV provirus integrated were also cloned from the RSV-NRK-2 cell line. The sequences of the regions involved in the recombination event have been determined and compared. Our data suggest that, compared with the sequence of viral DNA in the large circular form of unintegrated viral DNA, the provirus lacks two base pairs at each end and that the provirus is flanked by a six-base-pair direct repeat of cellular DNA. This six-base-pair repeat was apparently created during the integration event because this sequence was present only once at the integration site before the provirus was inserted. A survey of eight other independent RSV transformed rat cell lines demonstrates that, in agreement with earlier results, the RSV proviruses have entered different segments of rat cell DNA. We have also determined the sequence of a second virus DNA-host cell DNA junction from a second RSV-transformed rat cell line (RSV-NRK-4) and find that there are no obvious similarities between the two integration sites or between the integration sites and the termini of viral DNA. PMID- 6270671 TI - Three-dimensional structure of the complex of skeletal muscle actin and bovine pancreatic DNAse I at 6-A resolution. AB - The structure of rabbit skeletal muscle actin complexed with bovine pancreatic DNase I has been determined by x-ray crystallographic methods at 6-A resolution. The analysis was based on a new orthorhombic crystal form, space group P212121, with one complex in the asymmetric unit. Six isomorphous heavy-atom derivatives yielding an overall figure of merit of 0.72 have been used to calculate the electron-density map. Molecular models for actin and DNase I derived from this map have dimensions 67 X 40 X 37 A and 50 X 50 X 40 A, respectively. The actin molecule is elongated and consists of a larger and a smaller domain, each containing density regions resembling a central beta-pleated sheet surrounded by alpha-helices. The highest electron-density peak is found in the cleft between the two domains, perhaps indicating the bound ATP. Observed crystal contacts between actin molecules and a model for the F-actin filament are discussed. Two high-affinity Ca2+-binding sites which also bind Ba2+ have been located at the surface of the DNase I molecule. PMID- 6270672 TI - Proton and hydroxide ion permeability of phospholipid vesicles. AB - The apparent permeability of H+ through phospholipid bilayers was determined by measuring H+ efflux from large unilamellar phospholipid vesicles with internal space buffered at pH 4. The value obtained is about 10(-9) cm/sec at room temperature, five orders of magnitude lower than was recently reported for the combined permeability for H+ and OH- [Nichols, J. W. & Deamer, D. W. (1980) Proc. Natl. Acad. Sci. USA, 77, 2038-2042]. The apparent permeability measured in this way is the sum of contributions from the movement of H+ and of uncharged species (HCl or HNO3) in equilibrium with anions in the solution. There is evidence that the uncharged species make the dominant contribution and that the permeability coefficient for H+ per se is no larger than 5 X 10(-12) cm/sec. An attempt to measure OH- permeability by use of vesicles buffered at pH 10 did not give a conclusive result because the vesicle walls appeared to be damaged by exposure to this pH. An apparent permeability coefficient of about 10(-7) cm/sec was estimated for undamaged membranes. PMID- 6270673 TI - Insulin synthesis in a clonal cell line of simian virus 40-transformed hamster pancreatic beta cells. AB - A clonal hamster beta cell line (HIT) was established by simian virus 40 transformation of Syrian hamster pancreatic islet cells. Cytoplasmic insulin was detected in all cells by indirect fluorescent antibody staining, and membrane bound secretory granules were observed ultrastructurally. Acidified-ethanol extracts of HIT cell cultures contained hamster insulin as determined by radioimmunoassay, radioreceptor assay, and bioassay. One subclone at passage 39 contained 2.6 micrograms of insulin per mg of cell protein. [3H]Leucine-labeled HIT insulin and proinsulin were identical to islet-derived proteins when compared by NaDodSO4/polyacrylamide gel electrophoresis of immunoprecipitates. HIT cell insulin secretion was stimulated by glucose, glucagon, and 3-isobutyl-1 methylxanthine. Insulin secretion at optimal glucose concentration (7.5 mM) was 2.4 milliunits per 10(6) cells per hr. Somatostatin and dexamethasone markedly inhibited HIT insulin secretion. The HIT cell line represents a unique in vitro system for studying beta cell metabolism and insulin biosynthesis. PMID- 6270674 TI - In vitro synthesis and posttranslational uptake of cytochrome c into isolated mitochondria: role of a specific addressing signal in the apocytochrome. AB - Administration of the thyroid hormone 3,3,5'-triiodo-L-thyronine (T3) to rats leads to a marked increase in hepatic levels of mRNA for cytochrome c. Messenger RNA prepared from the free polysomes of T3-treated rats directed the in vitro synthesis of a polypeptide which only differed in amino acid sequence from mature cytochrome c in that it contained an NH2-terminal methionine. The in vitro product was incorporated specifically into purified rat liver mitochondria and became inaccessible to added trypsin when the mitochondria were added after translation was completed. Horse heart apocytochrome c, but not the holocytochrome, could compete with the in vitro synthesized polypeptide for its uptake into mitochondria. This suggests that the primary structural features of apocytochrome c, which serve as an addressing signal for mitochondria, are masked after the acquisition of heme and that this process occurs in the mitochondria. The addressing signal seems to be contained in a specific segment of the cytochrome polypeptide because only one fragment generated by CNBr cleavage of horse apocytochrome c, extending from residue 66 to the carboxy end of the molecule, could compete with the in vitro product for its transfer into mitochondria. PMID- 6270675 TI - Functional unit of the low density lipoprotein receptor of fibroblasts: a 100,000 dalton structure with multiple binding sites. AB - The low density lipoprotein (apoprotein B,E) receptors of fibroblasts bind plasma lipoproteins that contain either the B (apo-B) or E (apo-E) apoproteins. These include the low density lipoproteins (LDL) containing apo-B and certain high density lipoproteins containing apo-E (e.g., the cholesterol-induced apo-E HDLc). The same receptor binds both LDL and apo-E HDLc, but the apo-E HDLc bind with much higher affinity. This higher affinity is due to the binding of apo-E HDLc to multiple receptor sites. One possible structural model for this receptor is that each apo-B,E receptor possesses a single binding site. Thus, the multiple binding of apo-E HDLc would require the recruitment of four independent receptors to bind to a single apo-E HDLc particle. A second model is that each receptor unit possesses multiple binding sites capable of binding one apo-E HDLc particle or four LDL particles. This study characterizes the apo-B,E receptors in situ in the membranes of fibroblasts by radiation inactivation. This technique allows one to determine the functional size of the receptor on the basis of the amount of radiation required to inactivate the structure. The larger the molecular size of the structure, the less ionizing radiation (generated by a linear accelerator) that will be required to abolish the receptor--ligand interaction. The functional size of the apo-B,E receptor is 106,000 Mr as determined with both LDL and apo-E HDLc binding after radiation inactivation. Furthermore, the data derived from radiation inactivation and Scatchard analysis indicate that each apo-B,E receptor has multiple binding sites and that each functional receptor unit, capable of binding one apo-E HDLc or four LDL particles, has a molecular weight of 106,000. PMID- 6270676 TI - Biological activity of cloned retroviral DNA in microinjected cells. AB - Avian retroviral DNA molecules that had been cloned from infected cells by using recombinant DNA techniques were microinjected into either uninfected chicken embryo fibroblasts (CEF) or CEF transformed by the envelope glycoprotein deficient Bryan strain of Rous sarcoma virus [RSV(--)cells]. Retroviral DNA injected into RSV(--) cells directed transcription of envelope mRNA, which was then able to complement the RSV(--) env deficiency and promote the production of infectious transforming virus. The retroviral DNA also directed the production of fully infectious virus after injection into uninfected cells or RSV(--) cells. Virus production began within 3--4 hr after microinjections. When 100 DNA molecules per cell were injected, almost all injected cells produced infectious virus. As the number of injected molecules per cell was decreased, a corresponding decrease was observed in the number of cells that produced infectious virus. DNA injected into the cytoplasm was 1/50th to 1/10th as effective in virus production as DNA molecules injected into the nucleus. DNA molecules containing one or two tandem copies of the viral long terminal repeat were equally effective in virus production. PMID- 6270677 TI - Cyclic AMP: a mitogenic signal for Swiss 3T3 cells. AB - Addition of cholera toxin (100 ng/ml) to quiescent cultures of Swiss 3T3 cells acts synergistically with serum (2-4%), insulin, phorbol esters, epidermal growth factor, and fibroblast-derived growth factor to stimulate DNA synthesis. In the presence of insulin, cholera toxin caused a dose-dependent increase in cumulative [3H]thymidine incorporation into acid-insoluble material and in the intracellular cyclic AMP (cAMP) level. The dose--response curves for the two processes were similar. Furthermore, addition of 1-methyl-3-isobutylxanthine (15--500 microM) or of 4-(3-butoxy-4-methoxybenzyl)-2-imidazolidinone (5--100 microM), both of which are potent inhibitors of cyclic nucleotide phosphodiesterase which are potent inhibitors of cyclic nucleotide phosphodiesterase activity, stimulated DNA synthesis and increased cAMP levels in Swiss 3T3 cells. These compounds strikingly potentiated the effect of cholera toxin on DNA synthesis and on cAMP levels. When quiescent Swiss 3T3 cells were exposed to cholera toxin (100 ng/ml) and insulin at 10 micrograms/ml (4- to 7-fold increase in cAMP level) or to these agents and 1-methyl-3-isobutyl xanthine at 50 microM (35-fold increase in cAMP level), DNA synthesis began after a lag of 16 hr. These results indicate that cAMP acts as a mitogenic signal for Swiss 3T3 cells and differ from the widely held view that cyclic AMP inhibits the proliferation of fibroblast cells. PMID- 6270678 TI - Temperature-sensitive RNA polymerase mutants of a picornavirus. AB - Temperature-sensitive (ts) RNA polymerase mutants of a picornavirus are reported. Two foot-and-mouth disease virus (FMDV) mutants designated ts 22 and ts 115 have been characterized. As judged by isoelectric focusing, both have charge alterations in P56a, the FMDV RNA polymerase protein. Virus RNA synthesis in cells infected with the mutants is severely impaired at the nonpermissive temperature. RNA polymerase purified from baby hamster kidney cells infected with these mutants exhibits a marked ts transcribing activity in vitro. Spontaneous revertants of both mutants have P56a polypeptides that are indistinguishable from the parental proteins on the basis of charge. The revertants regain the ability to synthesize virus RNA in vivo at the nonpermissive temperature. RNA polymerase purified from the revertants remains transcriptionally active at the nonpermissive temperature. PMID- 6270679 TI - Specific transcriptional initiation in vitro on murine type C retrovirus promoters. AB - We have investigated the ability of molecularly cloned murine type C retroviral DNA to direct accurate initiation of RNA synthesis when added to cell-free extracts. Two different cloned proviruses were used. The first was derived from an integrated molecule of AKR murine leukemia virus and contains adjacent host information. The origin of the second was an unintegrated permuted copy of Harvey murine sarcoma virus. We found that the leukemia virus cloned provirus, as predicted by structural considerations, contained two functional RNA polymerase II promoters located in the U3 region present at either end of the molecule. These promoters initiate transcription at equal rates in vitro. We also found that the permuted sarcoma virus clone contained an RNA polymerase II promoter in the U3 region. Removal of viral sequences 49 bases upstream of the in vitro sarcoma virus initiation site by restriction cleavage results in loss of specific transcription, indicating a role for this information in in vitro promotion. The 5' ends of in vitro and in vivo viral RNA were compared by nuclease mapping techniques and found to be identical. Based on this evidence, we conclude that murine retroviral genomes contain sufficient information to initiate transcription independent of any host information in vitro and that these viral promoters are probably also active in vivo. In addition to the promoter in U3, Harvey murine sarcoma virus contains a second promoter in vitro that initiates near the 5' boundary of the transformation-specific (src) region of the virus. Initiation by this promoter was insensitive to low levels of alpha-amanitin, and the RNA transcript could be terminated to yield a 340-nucleotide product. PMID- 6270680 TI - Ubiquitous cell-surface glycoprotein on tumor cells is proliferation-associated receptor for transferrin. AB - A murine monoclonal antibody (OKT9) raised against human leukemic cells binds to a wide variety of leukemia and tumor cell lines and to a minority of leukemia cells taken directly from patients. Fetal thymus and liver are strongly reactive as are some normal, immature hemopoietic cells and activated lymphocytes. Reactivity with OKT9 appears to correlate with proliferation status in both normal and malignant populations. Biochemical analysis indicates that this structure is a approximately equal to 180,000-dalton glycoprotein with two disulfide-bonded subunits of approximately equal to 90,000-daltons. Isolation of the transferrin receptor from a T-cell line (MOLT-4) indicates that it also has a dimeric approximately equal to 180,000-dalton structure. Radio-labeled transferrin bound to its receptors can be specifically precipitated by the monoclonal OKT9, although the latter does not bind transferrin itself, indicating that the antigenic structure defined by this antibody is likely to be the transferrin receptor. PMID- 6270681 TI - Subunit structure of cell surface proteins: disulfide bonding in antigen receptors, Ly-2/3 antigens, and transferrin receptors of murine T and B lymphocytes. AB - The surface proteins of lymphocytes from spleen and thymus and several cultured lymphoid tumor lines were radioiodinated in situ, solubilized with Triton X-100, and examined for the presence of disulfide-bonded subunits by two-dimensional (intact, reduced) NaDodSO4/polyacrylamide gel electrophoresis. [Hynes, R. O. & Destree, A. (1977) Proc. Natl. Acad. Sci. USA 74, 2855-2859]. Few lymphocyte surface proteins were found to consist of disulfide-bonded subunits, and the most prominent of these could be identified. In normal B lymphocytes and B-lymphoma cells, IgD or IgM (or both) were the major disulfide-bonded proteins, and these were easily detectable, even without immunoprecipitation. In contrast, analysis of thymocytes and T-lymphoma cells did not reveal any protein resembling immunoglobulin in its chain structure. The major labeled thymocyte membrane protein consisting of disulfide-bonded subunits was identified as the Ly-2/3 antigen. It appeared to contain disulfide-bonded homodimers of Mr 35,000 (alpha 2) noncovalently associated with a second pair of homodimers of Mr 30,000 (beta 2). Peptide mapping showed these polypeptides to be homologous. A third disulfide bonded homodimer, which was heterogeneous in apparent Mr, appeared to be part of the Ly-2/3 complex. All cultured T- and B-lymphoma lines examined were found to possess a major surface protein that appeared to be a disulfide-bonded homodimer of a polypeptide of Mr 95,000. This protein was identified as the receptor for transferrin. It is suggested that the presence of two or more subunits in cell surface receptors renders their ligand functionally bivalent, making ligand induced receptor aggregation possible. PMID- 6270682 TI - Membrane attack complex of complement: distribution of subunits between the hydrocarbon phase of target membranes and water. AB - Membrane destruction by complement is effected by the membrane attack complex (MAC) which is the dimer of a fusion product of the complement proteins C5b, C6, C7, C8, and C9. Phospholipid bilayer vesicles were used as target membranes for the MAC and its intermediate complexes. The subunits of these membrane-bound complexes were explored as to their relative exposure to the hydrocarbon phase of the lipid bilayer and to water surrounding the lipid vesicles. Protein exposed to the aqueous phase was labeled with 125I; protein exposed to the hydrocarbon phase was labeled by using tritiated azido phospholipids and irradiation. Analysis of the membrane-bound MAC showed that subunits C5b, C8 beta, and C9 were exposed to the aqueous phase. The subunits C8 alpha-gamma and C9 were primarily in contact with the hydrocarbon phase. C6 and C7 were little exposed to either phase, suggesting that these proteins are inaccessible within the MAC. Analysis of the intermediate complexes showed that C5b was the subunit most exposed to water in membrane-bound C5b-7, and C5b and C8 beta were the water-exposed subunits in C5b 8. Subunit exposure to the hydrocarbon phase of the lipid bilayer changed during MAC assembly. Whereas all three subunits of C5b-7 carried the phospholipid photolabel; most of the label was bound to the C8 subunit in C5b-8 and to C9 in the MAC. It is proposed that contact with the hydrocarbon core of membranes is established by C5b-7 through each of its subunits, by C5b-8 through C8, and by the MAC through C8 and, particularly, C9. PMID- 6270683 TI - Evidence against extrapancreatic insulin synthesis. AB - Labeled and unlabeled insulin in acid/ethanol tissue extracts can be concentrated up to 100-fold by using a hydrophobic adsorption technique. After adsorption to and elution from an octadecylsilyl silica column, insulin is recovered in yields greater than 75%. By using this method of concentration, insulin in brain tissues of three of four fed rats and one rabbit was found to be less than 20% of plasma concentration. The kidney is the only extrapancreatic organ in which insulin is observed to be markedly above plasma levels. Porcine-insulin-like material was not detectable in guinea pig tissues (less than 0.02 ng/g). It is concluded that insulin is not synthesized in brain or other extrapancreatic tissues and that other mammalian insulins are not found in guinea pig tissues. PMID- 6270684 TI - Contractile activities of structural analogs of leukotrienes C and D: role of the polar substituents. AB - Twenty-three structural analogs of the leukotriene components of slow reacting substance of anaphylaxis (SRS-A), in which the polar regions of the leukotriene were systematically modified, were tested for their contractile activities on guinea pig pulmonary parenchymal strips and guinea pig ileum. The structural modifications allowed evaluation of the separate contributions of the four polar units in the C-1 to C-6 region of the SRS-A leukotrienes to smooth muscle spasmogenic activity. The free NH2-terminal amino group of the S-linked peptide was necessary for full activity, and its deletion or substitution reduced activity by more than one but less than two orders of magnitude. A similar level of importance was apparent for the free glycine carboxyl group. In contrast, a free eicosanoid carboxyl at C-1 is not required for full activity on the airway and for substantial activity on the ileum. A role for the C-5 hydroxyl is indicated by the inactivity of the one available 5-desoxy analog. Nucleophilic, divalent sulfur is not critical to leukotriene D (LTD) activity, in that one sulfoxide had substantial function. The conformational relationship between the eicosanoid and peptide moieties of LTD is of considerable importance in that epimers at the C-5 or C-6 position were less active than LTD by more than two orders of magnitude. Several lines of evidence suggest that the relative geometrical arrangement of the C20 chain and the peptide unit is important to activity, consistent with the existence of a true receptor for LTD. PMID- 6270685 TI - Monoclonal antibodies that demonstrate specificity for several types of human lung cancer. AB - Monoclonal antibodies with selectivity for human lung cancer were produced by immunizing BALB/c mice with an established line of human small cell lung cancer (NCI-H69) and fusing the mouse spleen cells to mouse myeloma line X63-Ag8.653. The resulting hybrid cells were initially screened by immunoautoradiography for production of antibodies that would react with NCI-H69 and another small cell lung cancer line (NCI-H128) but not its autologous B-lymphoblastoid line (NCI H128BL). Stable monoclonal antibody-producing lines were isolated by repeated cloning. Three independently derived monoclonal antibodies, designated 525A5, 534F8, and 538F12, were found to react with three of the major types of human lung cancer (small cell, adenocarcinoma, and squamous carcinoma). They did not react with bronchioloalveolar and large cell lung cancers, myeloma, lymphomas, leukemias, osteogeneic sarcoma, mesothelioma, hypernephroma, malignant melanoma, simian virus 40-transformed human fetal lung cells, skin fibroblast lines, human B-lymphoblastoid lines, human erythrocytes, and rodent cells. Interestingly, these antibodies also bound to three out of three human neuroblastomas and two out of three breast cancers but failed to react with mouse neuroblastoma and rat pheochromocytoma. The monoclonal antibodies reacted with human small cell lung cancer tumors obtained at autopsy, but had insignificant reactions with normal human lung, liver, spleen, and skeletal muscle. We conclude that monoclonal antibodies have been generated that react with common antigenic determinants expressed on several human lung cancer types, neuroblastoma, and some breast cancers, but are not detectable by our current assays on a variety of other human tumors or normal adult human tissues. Such antibodies are of potential clinical and biological importance. PMID- 6270686 TI - Monoclonal antibodies against beta nerve growth factor and their effects on receptor binding and biological activity. AB - Two hybrid cell lines, MC beta-1 and MC beta-2, secreting monoclonal antibodies against mouse submaxillary gland beta nerve growth factor (beta NGF), were produced by interspecies hybridization of spleen cells from rats immunized with beta NGF and mouse myeloma cells. The antibodies secreted by the two hybridomas are of the IgG1 subclass and bind staphylococcal protein A. The equilibrium dissociation constant of the beta NGF--antibody complex was determined for the MC beta-1 antibodies in solid phase and in solution. On protein A-coated surfaces the Kd is 3 X 10(-10) M, 2 orders of magnitude lower than the Kd 2 X 10(-8) M obtained in solution. The antigenic site recognized by MC beta-1 antibodies is present on each protomer of the beta NGF dimer, and the binding affinity of the second antibody molecule is similar to that of the first. The MC beta-1 antibodies inhibit neurite outgrowth from sensory neurons. Because this inhibition directly correlates with the inhibition of binding to the higher affinity beta NGF receptors, it suggests that beta NGF complexed with two antibody molecules does not bind to the receptor and is biologically inactive. PMID- 6270688 TI - Seesaw signal processing in pineal cells: homologous sensitization of adrenergic stimulation of cyclic GMP accompanies homologous desensitization of beta adrenergic stimulation of cyclic AMP. AB - Studies of the adrenergic regulation of cyclic GMP in the pineal gland show that (-)-norepinephrine stimulates cyclic GMP primarily in pineal cells, rather than in nerve endings as previously thought. The response exhibits the interesting and unusual characteristic of homologous sensitization: It is maintained by neural stimulation and disappears gradually as a consequence of depressed neural stimulation, due to denervation or decentralization of the superior cervical ganglia or to constant light. The response is restored in intact animals that had been in a constant-light environment when they are returned to a normal light cycle and in ganglionectomized animals by norepinephrine treatment. These findings are especially interesting because the pineal adrenergic--cyclic AMP stimulus--response system exhibits homologous desensitization. The occurrence of homologous sensitization of a cyclic GMP response and desensitization of a cyclic AMP response, which we term seesaw signal processing, in the same tissue or cell has intriguing implications. It provides a mechanism through which the qualitative nature of a multicomponent response can be modified. Such a mechanism could play a role in signal processing by neural or neuroendocrine tissues that release two or more extracellular messages. PMID- 6270687 TI - Purification of the saxitoxin receptor of the sodium channel from rat brain. AB - The saxitoxin (STX) receptor has been purified 740-fold from rat brain by a combination of ion exchange chromatography, wheat germ agglutinin chromatography, and sedimentation on sucrose gradients to a specific activity of 1488 pmol/mg of protein. The best fractions were estimated to be 47% pure from their specific activity or 66% pure on the basis of NaDodSO4 gel electrophoresis. Two polypeptides, alpha (Mr approximately equal to 270,000 +/- 10,000) and beta (Mr approximately equal to 38,300 +/- 2000) (mean +/- SD) copurify with STX binding activity. Two polypeptides of the same apparent Mr are specifically covalently labeled by photoreactive derivatives of 125I-labeled scorpion toxin in rat brain synaptosomes and are likely to be identical to alpha and beta. The solubilized STX receptor has a Mr of 316,000 +/- 63,000, limiting its composition to one alpha polypeptide and one or more beta polypeptides per soluble receptor. Our results suggest that the alpha and beta polypeptides contain both the STX binding site and the scorpion toxin binding site of the mammalian sodium channel. PMID- 6270689 TI - Characterization and partial purification of solubilized active opiate receptors from toad brain. AB - Opiate receptors have been solubilized from toad brain membranes in active form by using digitonin. Between 40% and 50% of the stereospecific binding activity present in toad brain membranes is recoverable in the ultracentrifugal supernatant of digitonin extracts. Binding of opiates to the solubilized receptor is enhanced 4- to 5-fold by decreasing digitonin concentration to 0.1% or less prior to binding. The solubilized receptor is similar to the membrane-bound receptor in its affinity for various ligands and its sensitivity to heat, trypsin, and N-ethylmaleimide. Moreover, the sodium effect seen in membrane-bound receptor is retained in the solubilized preparation. Both membrane-bound and soluble toad receptors show weak binding of enkephalins, suggesting that they are predominantly of the mu type. The solubilized opiate receptor has an approximate molecular weight of 350,000-400,000. Purification of up to 20-fold has been achieved by gel filtration on Sepharose CL-6B. PMID- 6270690 TI - Anatoxin-a interactions with cholinergic synaptic molecules. AB - Anatoxin-a, a bicyclic amine isolated from blue-green alga, binds to the nicotinic acetylcholine receptor of Torpedo electric tissue, thereby inducing conformational changes in the postsynaptic receptor--ion channel complex as evidenced by alterations in the binding of radiolabeled ligands to the complex. Anatoxin-a binds to the acetylcholine recognition site (Kd = 0.1--0.2 microM) as indicated by its competitive inhibition of specific [3H]acetylcholine and d [3H]tubocurarine binding, Anatoxin-a stimulates the binding of three physiologically identified "ion channel blockers," [3H]perhydrohistrionicotoxin, [3H]phencyclidine, and [3H]phencyclidine methiodide. The 50% effective doses for these effects range from 0.14 to 0.28 microM. Incubation of Torpedo membranes with anatoxin-a before addition of a radiolabeled channel probe produces a time- and concentration-dependent attenuation of the binding compared to the situation in which anatoxin-a and the probe are added simultaneously. The time course for the elaboration of this decrease corresponds to electrophysiological measurements of anatoxin-a-induced desensitization of neuromuscular junction responses. In these nicotinic actions, anatoxin-a is about as potent as acetylcholine. Anatoxin a has relatively low affinity for the muscarinic acetylcholine receptors of rat brain, inhibiting 3-[3H]quinuclidinyl benzilate binding (10(-10) M) by 50% at concentrations between 10 and 20 microM. In contrast to classical muscarinic agonists, anatoxin-a displays little regional selectivity in its binding, and its receptor affinity is unaltered by alkylation of the neural membranes with N ethylmaleimide. PMID- 6270692 TI - Phosphorylation and dephosphorylation as a mechanism for metabolic control within the red blood cell. PMID- 6270691 TI - Immunotherapy with dialyzable leukocyte extracts and studies of their antigen specific (transfer factor) activity. PMID- 6270693 TI - The hematology and biochemistry of canine postnatal anemia. AB - In conclusion, postnatal changes in the 2,3-DPG concentration of the canine erythrocyte are directly related to changes in erythrocyte 1,3-diphosphoglycerate concentration. There is not apparent differential regulation of 2,3-DPG mutase or phosphatase activities. Glucose consumption and lactate production of the canine erythrocyte do not change during postnatal development, thus, overall rate controlling reactions in the upper portion of the glycolytic pathway do not regulate the changes in 1,3-diphosphoglycerate. The analysis of intermediate and enzyme levels below the 2,3-DPG shunt indicates that pyruvate kinase functions as a controlling sink reaction. Variation in the pyruvate kinase step causes changes in the erythrocyte phosphoenolpyruvate levels. Since the enolase, phosphoglyceromutase and phosphoglycerate kinase reactions are at equilibrium, changes in phosphoenolpyruvate concentrations are paralleled by changes in the concentrations of 1,3-diphosphoglycerate and 2,3-DPG. We propose that the rise in erythrocyte 2,3-DPG, during the first 60 days of postnatal age, results from a decline in the levels of the fetal or M2 isozyme of pyruvate kinase. The subsequent decline in erythrocyte 2,3-DPG to normal adult levels may result from a developmental change in pyruvate kinase isozymes. The changes in erythrocyte glycolysis following birth increase the oxygen transporting efficiency of hemoglobin at the higher oxygen tensions of the neonatal environment. This provides an initial reserve of blood oxygen transport capacity which suppresses erythrocyte production, resulting in the condition defined as postnatal anemia. There is no apparent defect in postnatal erythropoiesis. PMID- 6270694 TI - In vitro influences of adrenaline on erythrocyte metabolism and on oxygen affinity of hemoglobin. PMID- 6270695 TI - Red cell anion channel blockade: extracellular modulation of internal membrane function. PMID- 6270696 TI - Cyclic AMP metabolism in P. berghei infected murine red cells. PMID- 6270697 TI - Hemoglobin and the red cell membrane: increased binding of polymorphic hemoglobins and measurement of free radicals in the membrane. PMID- 6270698 TI - De novo analysis of alpha-aminoalkanesulfonic acids. PMID- 6270699 TI - Lithium increases selective attention in rats. AB - Both 0.15 and 1.50 mEq/kg lithium chloride were found to increase inspective exploration (i.e. behaviors directed at the close examination if discrete stimuli in a new environment) in rats. In addition, 1.50 mEq/kg LiCl decreased general activity, irrespective of the characteristics of the experimental situation. The dose-independent increase in inspective exploration was interpreted as increased selective attention to salient stimuli which provide information about the environment. This hypothesis predicts that both doses of lithium should decrease distractibility by irrelevant stimuli. This prediction was confirmed in a second experiment, in which the ability of a tone to suppress drinking by thirsty rat was reduced by both doses of lithium. A two-factor model of lithium's action in rats is proposed. Lithium produces a dose-independent improvement of selective attention to stimuli which provide detailed information about the environment. In addition, lithium exerts dose-related effects on activity levels: a mild increase in activity with dose in the 0.15-0.20 mEq/kg range and a decrease in activity with doses of 0.50 mEq/kg and higher. PMID- 6270701 TI - Cholesterol, bile acid and bile salt adsorption to bran in vitro. PMID- 6270700 TI - Effects of polyphloretin phosphate (PPP) and quinterenol of cyclic nucleotide phosphodiesterase activities. PMID- 6270702 TI - Effect of theophylline and imidazole on cyclic AMP and cyclic GMP levels in active sartorius muscle of Rana esculenta. PMID- 6270703 TI - Alpha and beta sympathetic responses to isoproterenol by the isolated rat vas deferens. PMID- 6270704 TI - Effect of converting enzyme and angiotensin II inhibitors in chronic one kidney bipolar ligated hypertension. PMID- 6270706 TI - Cardiovascular responses to the intravenous and the intracerebroventricular administration of lithium chloride. PMID- 6270705 TI - Cyclic AMP mediator of gastric mucosa cytoprotection during fatty acids damaging. PMID- 6270707 TI - Plasma viscosity: relation to age, atherogenic diet and suloctidil. PMID- 6270708 TI - [Effects of synthetic neuropeptides in psycho-organic brain syndrome. Results with ACTH4-10 and ACTH4-9-analog (author's transl)]. AB - The effects of ACTH4-10 (Org O163) and of the ACTH4-9 analog (Org 2766) were tested in 20 male patients suffering from a mild to moderate cerebroorganic impairment. Patients were aged between 51-72 years (mean 60.9). Org 2766 was given in dosages of 0.05 mg, 0.5 mg and 5 mg, Org O163 in a dosage of 30 mg. The investigation was based on a randomized incomplete crossover design. By means of psychological tests the effects of the synthetic neuropeptides on memory, state of well-being, attention, vigilance and psychomotor function were investigated. The statistical analysis of the results did not reveal any effects of both substances on the functions tested. These results are in agreement with those of other studies in man using similar methods and acute administration of ACTH4-10 and/or Org 2766. In such studies only effects of reactive inhibition of attention and motivation could be demonstrated consistently. PMID- 6270709 TI - Ethanol increases opioid activity in plasma of normal volunteers. AB - The effect of acute ethanol administration on plasma levels of beta-endorphin immunoreactivity and opioid activity was measured in 4 normal volunteers. 60 min following ethanol consumption opioid activity levels, measured by radioreceptorassay, increased significantly with peak rises of more than 400%; levels of beta-endorphin-immunoreactivity did not change significantly. These results are compatible with the effect of the opiate-antagonist naloxone, reversing ethanol-induced coma. PMID- 6270710 TI - Kinetics of drug action and equilibrium results at the node of Ranvier. PMID- 6270711 TI - Junctional intercellular communication: the cell-to-cell membrane channel. PMID- 6270712 TI - Enhanced acquisition and retention of passive avoidance behaviour in adult rats given a single transuterine ACTH injection while fetuses. AB - The long-term effects of a single ACTH injection administered to fetuses by transuterine way, on the ability to learn and retain a noxious significance were studied. The investigation was carried out on 115 adult rats of either sex. During the experiment the authors examined the range of inborn avoidance behaviour (light avoidance), as well as the ability to develop and retain a passive avoidance response. The ACTH-treated group did not present a significant difference of light avoidance behaviour as compared to controls, showing nevertheless a significantly higher learned passive avoidance and a delayed extinction of the noxious significance. The role of pituitary-adrenal hormones in the mechanism of avoidance behaviour during ontogenetic development is discussed. PMID- 6270713 TI - Buried microvascular free flaps for reconstruction of soft-tissue defects. PMID- 6270714 TI - "Staying power"--absorbable vs. nonabsorbable. AB - An experiment was designed to compare the "holding power" or "staying power" of absorbable (polyglycolic acid and polyglactin 910) and nonabsorbable (nylon) suture. The aim of this experiment was to determine what provides the lasting strength of the bond between soft tissues that are approximated or plicated. When correcting the rectus diastasis during abdominoplasty, we used nylon sutures in 15 patients and absorbable synthetic sutures in 15 other patients. We then marked the closed folds of the rectus sheath with small metal vascular clips. Two days later and approximately 6 months after operation an upright anteroposterior abdominal x-ray was taken and the position of the metal clips was compared in the test groups. Although there was usually slight separation of the clips after 6 months, no significant difference between the two groups was noted, thereby indicating that holding power is not related to type of suture material but more likely to fibroplasia. PMID- 6270715 TI - [Lung cancer in Slovenia and its epidemiological trends. (Study for the period from 1953 to 1976) (author's transl)]. PMID- 6270716 TI - The clinical approach to the evaluation of diarrhea. AB - Diarrhea may present as an acute transient condition that is hardly more than an inconvenience or as an acute devastating condition that decimates whole armies and cities. In places in which it is endemic, chronic diarrhea may be equally devastating; however, chronic diarrhea is usually the problem of an individual patient. A clinician may require the most extensive analysis for proper appraisal of chronic diarrhea. A thoughtful, thorough history and physical examination will often serve to narrow the number of tests required to reach a satisfactory diagnosis. Rational therapy may then be quickly determined from many existing references. PMID- 6270717 TI - Studies on the disposition of Li+ in the guinea-pig and rat. AB - Rats and guinea-pigs were given LiCl acutely (2 mmol/kg IP or intragastrically) or chronically (daily doses 0.6-4 mmol/kg) and plasma, erythrocytes, kidney, liver and brain were analysed for Li+. Generally, after acute LiCl, tissue Li+ levels followed changes in plasma Li+ levels. However, brain Li+ concentrations changed more slowly and in the rat, but not in the guinea-pig, paralleled erythrocyte Li+ concentrations. Li+ was absorbed more slowly from the gastrointestinal tract of the rat. After chronic LiCl, the erythrocyte:plasma Li+ ratio was about 0.1 in the guinea-pig and about 2 in the rat. Relative Li+ tissue concentrations were as follows: guinea-pig, plasma greater than kidney greater than liver greater than brain greater than erythrocyte;rat much greater than erythrocyte = brain greater than liver = plasma. A Na+-dependent Li+ efflux was demonstrated in the erythrocytes of the guinea-pig and human, but not the rat. This process was inhibited by phloretin (0.2 mM), but not frusemide (2.0 mM). The marked differences in the activity of the erythrocyte Na+-dependent Li+ countertransport process in the guinea-pig and rat could extend to other tissues and explain the observed interspecies differences in tissue Li+ distribution. PMID- 6270718 TI - Methods for evaluating effects of opioid peptides in laboratory animals. PMID- 6270719 TI - Drug treatment and family intervention during the aftercare treatment of schizophrenics. PMID- 6270720 TI - [Free-radical formation in biomacromolecules as affected by neutrons and gamma radiation]. PMID- 6270721 TI - [Effect of a neurodystrophic process on cAMP phosphodiesterase activity in the myocardium and gastrocnemius muscle in the irradiated rabbit]. PMID- 6270722 TI - [Neuronal injuries of the rat cerebral cortex after exposure to protons of varying energies]. PMID- 6270723 TI - [Radiodiagnostic procedure in congenital arterio-venous malformations (author's transl)]. PMID- 6270724 TI - Further observations on the arthropathy of calcium pyrophosphate crystal deposition disease. AB - The arthropathy of calcium pyrophosphate dihydrate (CPPD) crystal deposition disease is distinctive and may affect lumbar spinal and sacroiliac joints, as well as appendicular joints. Subchondral pseudocysts that are a hallmark of the disease have a variable appearance, but often occur as a typical cluster of subchondral, coalescent lucencies with smudged, sclerotic margins. Structural joint collapse with fragmentation of cartilage and bone may occur and appear to be related, at least in some cases, to antecedent pseudocysts. Characteristic intra-articular osteochondral bodies are often extensive and may affect multiple joints; their pathogenesis is discussed. Articular synovial calcification is common and may be due to calcium hydroxyapatite, as well as CPPD, particularly if advanced degenerative changes are present. Recognition of the radiologic features may be encountered in CPPD crystal deposition disease is important for differential diagnosis. PMID- 6270725 TI - Ultrasonography of alcoholic liver disease with histological correlation. AB - In alcoholic liver disease (fatty infiltration, alcoholic cirrhosis), the liver is diffusely abnormal on ultrasound. Changes in size, dilatation of the hepatic veins, and ascites may also occur. The authors conducted a histological correlation of these abnormalities in 22 alcoholic patients and 16 controls, grading the changes on a scale of 0 to 4+ for fat, fibrosis, and necrosis and noting tumor whenever present. Ultrasound detected abnormality in 21 cases (sensitivity = 95%) and correctly identified 15 controls (specificity = 94%). Of the 5 tumors seen, 4 hepatomas were detected and biopsied and 1 metastatic squamous-cell carcinoma was missed. Applications of commercially available A-scan module are considered and its limitations discussed. With the exception of minimal change (1+ fat or fibrosis), ultrasound detected many of the pathological changes seen in alcoholic liver disease. PMID- 6270726 TI - A noninvasive method for measuring portal venous/total hepatic blood flow by hepatosplenic radionuclide angiography. AB - Radionuclide angiography was used to generate first-pass radioactivity vs. time curves for the left heart, right hepatic lobe, right lung, spleen, and both kidneys following rapid intravenous injection of 20 mCi (740 MBq) of 99mTc pertechnetate. Seven normal subjects were examined as well as 57 cirrhotic patients, who also underwent angiographic grading of portal venous perfusion. For analysis, two time points were identified: (a) t0, when 99mTc first entered the liver (the initial rise of either curve); and (b)tc, when 99mTc was maximal in abdominal organs (the renal peak). Analysis was based on the slopes of the two phases of the hepatic curves t0 + 7 seconds and Tc + 7 seconds; this time selection permitted analysis of all curves. The hepatic perfusion index (HPI) = slope (tc + 7 secs)/slope (t0 + 7 secs) + slope (tc + 7 secs). The mean HPI for the normal subjects was 66% +/- 7; for the cirrhotic patients with angiographic Grades I, II, III, and IV, the HPI was 52% +/- 9, 37% +/- 6, 15% +/- 7, and 3% +/ 4, respectively. Correlation between HPI and angiography was significant (p less than 0.001). This method offers a readily available, rapid, relatively inexpensive, and quantitative method of grading the ratio of portal venous to total hepatic blood flow. PMID- 6270728 TI - The Drash syndrome: male pseudohermaphroditism, nephritis, and Wilms tumor. AB - We report the radiographic and clinical findings of five infants and children who had the combination of male pseudohermaphroditism, progressive nephritis, and Wilms tumor. The components of this syndrome are so striking that the radiologist can often suggest the diagnosis months to years before all three components are apparent, if proteinuria, hypertension, or an abdominal mass develop in any child with male pseudohermaphroditism. Since the progressive renal failure probably can be managed by dialysis or renal transplantation in some cases, prompt recognition and treatment of the Wilms tumor may permit the child to live a long and relatively normal life. PMID- 6270727 TI - 97Ru-DMSA for delayed renal imaging. AB - Dimercaptosuccinic acid (DMSA) was labeled with 97Ru both with and without the addition of SnCl.2H2O. The tin-containing preparation was found to induce higher cortical deposition of 97Ru-DMSA than the tin-free preparation. Visualization of the renal cortex was excellent 4 to 48 hours after injection in normal dogs with renal insufficiency. It is concluded that 97Ru-(Sn+2)-DMSA is a potentially useful renal imaging agent when delayed scintigraphy is necessary because of decompensaton of the kidneys. PMID- 6270729 TI - Efficacy of whole lung tomography in diagnosing metastases from solid tumors in children. AB - Whole lung, frontal tomography is compared with frontal chest radiography in the diagnosis of pulmonary metastasis in children with cancer. Of 79 patients studied, 65 had a total of 195 routine tomograms. In only 2.7% of studies did tomography yield new information, and in only 1% was an additional lesion visualized. In 27 patients, tomography was performed after an abnormality was seen on the chest radiograph. New information was obtained in 15 (32%) of those 47 studies; additional nodules were identified on six occasions (four metastatic, two inflammatory), but only once was treatment altered. In one study, tomography localized a nodule seen only on a lateral chest radiograph, and in eight studies it excluded metastasis following an abnormal frontal chest radiograph. PMID- 6270730 TI - Ventricular dysrhythmia: membrane basis or of currents, channels, gates, and cables. PMID- 6270731 TI - Cellular electrophysiology of ventricular and other dysrhythmias: studies on diseased and ischemic heart. PMID- 6270732 TI - Decoding the pattern of protein evolution. PMID- 6270733 TI - [Changes in amino acid and sugar transport activity associated with malignant transformation (author's transl)]. PMID- 6270734 TI - [Energy (ATP) reproduction and its application by bioreactor (author's transl)]. PMID- 6270735 TI - [Involvement of glucocorticoid and epidermal growth factor in the development of palate in human embryo (author's transl)]. PMID- 6270736 TI - Dissociation of beta-adrenergic stimulation of renin secretion and prostacyclin synthesis in the rabbit kidney. AB - The effect of inhibition of prostaglandin (PG) synthesis with indomethacin on basal and isoproterenol-stimulated renin secretion was examined in the isolated perfused rabbit kidney. 6-keto PGF1 alpha' the stable metabolite of prostacyclin, was measured in urine by radioimmunoassay using 125I labelled histamine coupled to 6-keto PGF1 alpha as ligand. The level in urine, prior to isolation and perfusion of the kidney, was 10.7 +/- 5.6 ng/min, and this was reduced to 0.32 +/ 0.25 ng/min (P less than 0.05) in rabbits treated with 2.0 mg/kg of indomethacin. Renin release was markedly stimulated by intrarenal infusion of isoproterenol (0.1 microgram/min) but urinary 6-keto PGF1 alpha did not change. These responses were not affected by indomethacin treatment. Renal perfusion pressure, perfusate flow rate and consequently renal vascular resistance, remained relatively constant during the course of perfusion and were unaltered by indomethacin treatment. These results therefore do not support a role for PGs, and in particular prostacyclin, in the renin response to beta-adrenergic stimulation with isoproterenol. PMID- 6270737 TI - Effect of PGE1 on PGF2 alpha-induced luteolysis in nonbred ewes. AB - Fifty-six ewes were used to study the effects of PGE1 or PGE2 plus PGF2 alpha given into the perivascular space of the ovarian vascular pedicle on luteal function of nonbred ewes. All ewes receiving PGF2 alpha had reduced levels of plasma progesterone and unoccupied receptors for LH at 24 hr after treatment regardless even if they received PGE1 or PGE2 concomitantly. Levels of plasma progesterone in ewes receiving only PGF2 apha were reduced further at 48 hour. Plasma progesterone and unoccupied receptors for LH of ewes receiving PGE2 + PGF2 alpha were maintained at 48 hr at levels seen at 24 hr after treatment, while progesterone in ewes receiving PGE1 + PGF2 alpha at 48 hr returned to levels seen in controls at 48 hr and unoccupied receptors for LH were three fold greater than controls. PMID- 6270738 TI - The effect of prostaglandin synthetase inhibitors on renal function: studies in normal rats during sodium or water diuresis and in rats with chronic renal insufficiency. AB - The effect of acute infusion of the prostaglandin synthetase inhibitors - meclofenamate or indomethacin - was examined in awake rats. Studies were performed in normal rats undergoing either sodium or water diuresis and in salt replete rats with chronic renal insufficiency. Prostaglandin synthetase inhibitors had no effect on renal plasma flow, glomerular filtration rate or fractional excretion of sodium in any of the groups. Absolute urinary excretion rates for sodium and potassium decreased only in the normal, salt-replete rats. In contrast, prostaglandin synthetase inhibitors consistently decreased urinary flow and osmolar clearance under all experimental conditions studied. In the normal, salt-replete rats the fall in urine flow was preceded by an increase in urinary excretion of cyclic AMP. These results show that inhibitors of prostaglandin synthesis enhance the ability of the kidney to reabsorb water. This effect may be secondary to increased cyclic AMP generation and to increased urea recirculation resulting in higher urea accumulation in the renal medulla. PMID- 6270739 TI - Prostacyclin stimulates the adenylate cyclase system of human thyroid tissue. AB - Since Prostacyclin (PGI2) is a major product of arachidonic acid metabolism in the human thyroid, we have studied the effects of PGI2 on cAMP accumulation in human thyroid slices and cultured thyrocytes. In both systems, PGI2 caused a dose and time-dependent increase of cAMP accumulation with higher potency and efficacy than PGE2. Two optically active isomers of 5,6-dihydro-PGI2, i.e. stable synthetic analogs of PGI2, had qualitatively similar effects to PGI2. The relative potency ratio between the alpha- and beta- isomer as well as their potency compared to PGI2 were substantially similar to their potency in inhibiting human platelet aggregation. In thyroid slices, PGI2 and its stable analogs had a greater effect than TSH in causing cAMP accumulation; however, in contrast to TSH, this effect was not associated with increased iodothyronine release except at maximal PGI2 concentrations. TSH had no detectable effect on thyroidal PGI2 synthesis and release. In cultured thyrocytes the effects of PGI2 and its stable analogs were considerably less than those obtained with TSH and required higher concentrations. Such a discrepancy was not found in the case of PGE2. These findings suggest the existence of a specific PGI2-responsive adenylate cyclase system in human thyroid cells other than thyrocytes, of possible physiologic significance. PMID- 6270740 TI - Prostaglandin E2 elevation of cyclic-AMP in granuloma macrophages at various stages of inflammation: relevance to anti-inflammatory and immunomodulatory functions. AB - In the carrageenin-induced granuloma of rats the inflammatory tissue growth and macrophage invasion on the one hand and the cyclic-AMP content of the macrophages on the other, display opposite directional changes. Macrophages, isolated from this tissue at different stages of inflammation, were used to examine the effect of prostaglandin E2 on intracellular levels of c-AMP. It appears that during infiltration of the macrophages into the inflammatory tissue, the sensitivity of adenylate cyclase to activation by PGE2 increases. Arguments are presented that these observations made in vitro, are in direct relevance to the previous described anti-inflammatory effect of PGE on granuloma tissue in vivo. PMID- 6270741 TI - Effects of prostaglandin E1, I2 and isoproterenol on the tissue cyclic AMP content in longitudinal muscle of rabbit intestine. AB - Effects of prostaglandin E1(PGE1) and prostaglandin I2(PGI2) on the mechanical activity and tissue cyclic AMP content of the longitudinal muscle of rabbit intestine were examined, comparing that of the tissue cyclic AMP content. Isoproterenol caused a relaxation and increased tissue cyclic AMP content. PMID- 6270742 TI - Leukotriene B4: an inflammatory mediator in vivo. AB - Leukotriene B4 (LTB4 isomer III), which promotes the movement and aggregation of leucocytes in vitro also stimulates the chemo-attraction of leucocytes and their adherence to vascular endothelium in vivo. These effects were observed directly in the hamster cheek pouch preparation and on histological examination of sections from the rabbit mesentery. Intravenous injection of LTB4 (isomer III) into the rabbit resulted in a profound but transient neutropenia. Intradermal injection of LTB4 (isomer III) in the rabbit produced a rapid accumulation of neutrophils and this effect was also observed in skin chambers applied over abrasions on the rabbit back or the human forearm. PMID- 6270743 TI - Stimulus-specificity of the chemotactic deactivation of human neutrophils by lipoxygenase products of arachidonic acid. AB - Preincubation of human neutrophils with chemotactic concentrations of 5(S) hydroxy-eicosatetraenoic acid (5-HETE) or 5(S), 12(R)-dihydroxy-6,14 cis-8,10 trans-eicosatetraenoic acid (leukotriene B4) induces a state of preferential chemotactic unresponsiveness to the homologous factor, termed deactivation, and less suppression of the responses to other chemotactic stimuli. The ratio of the concentrations required for maximal chemotactic deactivation of neutrophils to that which stimulates chemotaxis optimally is greater for 5-HETE and leukotriene B4 than for peptide and protein factors. In contrast to other chemotactic factors, 5-hydroperoxy-eicosatetraenoic acid (5-OOHETE) induces neutrophil chemotactic deactivation that is independent of the nature of the subsequent stimulus and is more slowly reversible after elimination of the fluid-phase deactivating factor. The unique characteristics of the chemotactic deactivation of human neutrophils by 5-OOHETE may be attributable in part to its endogenous metabolism to potent deactivating factors or to covalent derivatization of subcellular structures of the neutrophils by the highly reactive 5-OOHETE. PMID- 6270744 TI - 9 alpha-homo-9,11-epoxy-5,13-prostadienoic acid analogues: specific stable agonist (SQ 26, 538) and antagonist (SQ 26, 536) of the human platelet thromboxane receptor. AB - A newly synthesized 9 alpha-homo-9,11-epoxy-5,13-prostadienoic acid analogue, SQ 26, 536, (8(R)9(S)11(R)12(S)-9 alpha-homo-9,11-epoxy-5(Z), 13(E)-15S hydroxyprostadienoic acid) inhibited arachidonic acid (AA)-induced platelet aggregation with an I50 value of 1.7 microM. SQ 26,536 did not inhibit prostaglandin (PG) synthetase activity of bovine seminal vesicle microsomes or thromboxane (Tx) synthetase activity of lysed human blood platelets. SQ 26,536 also inhibited platelet aggregation induced by epinephrine (secondary phase), 9,11-azoPGH2 and collagen but did not inhibit the primary phase of epinephrine induced aggregation or ADP-induced platelet aggregation. SQ 26,538 (8(R)9(S)11(R)12(S)-9 alpha-homo-9,11-epoxy-5(Z),13(E)-15R-hydroxyprostadienoic acid), a 15-epimer of SQ 26,536, induced platelet aggregation with an A50 value of 2.5 microM. SQ 26,536 competitively inhibited SQ 26,538-induced platelet aggregation with a Ki value of 3 microM. Neither indomethacin, a PG synthetase inhibitor, nor SQ 80,338 (1-(3-phenyl-2-propenyl)-1H-imidazole), a Tx synthetase inhibitor, inhibited SQ 26,538- or 9,11-azoPGH2-induced platelet aggregation. These data indicate that SQ 26,536 and SQ 26,538 are stable antagonist and agonist, respectively, of the human blood platelet thromboxane receptor. PMID- 6270745 TI - [Differential diagnosis between hereditary fructose intolerance and fructose-1,6 diphosphatase deficiency]. PMID- 6270746 TI - [Clinical incidence of bovine brucellosis in the north Ivory Coast]. PMID- 6270747 TI - [Note on a trypanocidal treatment trial in the control of primary infection in Baoule calves]. PMID- 6270748 TI - [Use of body measurements to estimate liveweight in local taurine breeds of the Ivory Coast]. PMID- 6270749 TI - [Milking and milk production in sedentary village herds in the north Ivory Coast]. PMID- 6270750 TI - [Cardiac gammagraphy with technetium pyrophosphate: positive image of acute myocardial infarct]. PMID- 6270751 TI - [The concept of ventilation-perfusion. A model for teaching]. PMID- 6270753 TI - [Inter- and intra-regional factors influencing the distribution of ventilation perfusion ratios]. PMID- 6270754 TI - [Distribution of ventilation-perfusion ratios using the inert gases method]. PMID- 6270752 TI - [Distribution of ventilation/perfusion ratios]. PMID- 6270755 TI - Nutritional aspects of bread and flour. Report of the Panel on Bread, Flour and other Cereal Products, Committee on Medical Aspects of Food Policy. PMID- 6270756 TI - [Mode of action of catecholamine on the myocardial cells (author's transl)]. PMID- 6270757 TI - [Coronary vasospasm and function of adrenoceptors (author's transl)]. PMID- 6270758 TI - [Cervico-uterine amebiasis. Characteristics of 15 cases and review of the literature]. PMID- 6270759 TI - [Preliminary study of Azolla sp., an aquatic fern with agronomic potential in the humid tropics of Mexico]. PMID- 6270760 TI - [New method of detecting gelatinase activity combined with tests for nitrates or MR-VP]. PMID- 6270761 TI - [Cytomegalovirus infection in healthy adults. Clinical, anatomo-pathological, hepatic and virological study of 6 cases]. PMID- 6270762 TI - [The VIP peptidergic system. Role in pathology and therapeutics (author's transl)]. PMID- 6270763 TI - [Ionic movements in pancreatic islet cells (author's transl)]. PMID- 6270764 TI - [Viruses and cancer (author's transl)]. PMID- 6270766 TI - [Primary hyperparathyroidism (author's transl)]. PMID- 6270765 TI - [Calmodulin, an intracellular calcium mediator (author's transl)]. PMID- 6270767 TI - [Morphological variations of bronchopulmonary cancers]. PMID- 6270768 TI - [Dysautonomia and behavioural disorders from periventricular metastasis (author's transl)]. AB - In a patient presenting dysautonomia a severe orthostatic hypotension revealed the presence of a subependymal metastatic infiltration from a large cell anaplastic bronchial carcinoma. Clinical pharmacological studies were conducted. A paradoxical sinus bradycardia during orthostatism, and an absence of bradycardia after clonidine injection, were suggestive of a central origin of the dysautonomia. At post-mortem, metastatic infiltration involved the walls of the lateral ventricles, and the floor of the IIIrd and IVth ventricles, from where it invaded structures responsible for cardiovascular control. Correlations could be established between the results of the clinical pharmacological tests and the pathological findings. Also present were a) anterograde amnesia dating from the onset of the orthostatic hypotension, probably due to compression of the anterior pillars of the trigone by tumour invasion of the septum; b) behavioural disorders with absence of spontaneous movement and speech, indifference and docility, probably arising from destruction of the septum. PMID- 6270769 TI - [Atypical ceroid-lipofuscinosis in the adult or atypical spinocerebellar degeneration with intracytoplasmic inclusions? (author's transl)]. PMID- 6270770 TI - [Diagnosis of bronchopulmonary cancer by cytological examination of the sputum. Cytohistological evaluation]. PMID- 6270771 TI - [Correlations between conventional allergometry and the Palmer types in interpretation of tuberculin skin tests]. AB - In view of differentiating post-vaccinal allergy from that developing after tuberculosis infection, the authors have investigated comparatively intradermal reactions to tuberculin according to classical allergometry, and to the 4 types described by Palmer-Edwards in a group of 1549 students and 211 patients. All the subjects were tested with 2 units of PPD IC 65. It was noted that the type I and II Palmer reactions corresponded, in the majority of the cases, to intense, or hyperergic ID reactions, while the types III and IV of Palmer corresponded to slightly positive, or to negative ID reactions. The presence of the post-vaccinal scar does not appear to influence significantly the type of the reaction, but the presence of active specific lesions is accompanied by reactions of type I and II of the Palmer classification. It is concluded that although the Palmer types do not allow to make a clear differentiation between the two variants of allergy they provide however useful indications for the interpretation of IDR in a more sophisticated manner, as compared with the conventional allergometry. PMID- 6270772 TI - [Improving the Lowenstein-Jensen medium]. AB - The authors have carried out some modifications of the formula of the Lowenstein Jensen medium in view of a better cultivation of the Koch bacilli. The changes consisted in the elimination of glycerin, addition of glucose, or sodium pyruvate and glucose, or in the addition of fractions of cultures of mycobacteria. The material seeded on this medium consisted in 20 wild strains of tubercle bacilli (M. tuberculosis) showing both drug sensitivity and drug resistance, and 167 samples of sputum obtained from patients with confirmed tuberculosis or from subjects suspected of tuberculosis. The possibility to identify the effects of sodium pyruvate and of glucose by increasing their concentration to 0,5 g% was also studied with another 68 products. The results demonstrated that addition of 0,25 g% of sodium pyruvate, and of 0,25 g% of glucose, as well as elimination of glycerin from the Lowenstein-Jensen medium stimulated the development of the Koch bacilli from pathological products, and increased the frequency of positive cultures. The development of colonies 1 to 2 weeks earlier and the obtention of more numerous eugenic colonies. PMID- 6270773 TI - [Difficult diagnoses in pneumology]. PMID- 6270774 TI - [Evolution of serum parameters of iron metabolism and of the natural resistance of tuberculosis patients under treatment]. AB - The authors have studied the evolution of serum parameters of iron metabolism in patients with tuberculosis during the standard chemotherapy. The results are discussed, in the perspective of a model of natural resistance depending on iron. No strict correlation was found between the bacteriostatic capacity of the serum and the process of saturation with iron of transferrin, probably due to participation in the bacteriostatic effect of other factors beside transferrin. The results have demonstrated that the imbalance between the serum values of copper (that are increased), and those of iron (that are low) tends to become normal during the recovery, as evaluated by a negativation of the sputum cultures. These results suggest that the use of a ratio between copper and iron in the blood serum as a method for assessing the recovery of the balance in the tuberculous organism during therapy is justified. PMID- 6270775 TI - [Current status of tuberculosis in the Socialist Republic of Romania and the prognosis for the next 5-year period (general report)]. PMID- 6270776 TI - [Short-term anti-tubercular chemotherapy in the light of current trends]. PMID- 6270777 TI - The etiologic role of M. bovis in the pulmonary and urogenital tuberculosis. PMID- 6270778 TI - An ultrastructural and cytoenzymological study on the folliculo-papillary thyroid carcinoma. AB - Authors studied ten cases of folliculo-papillary thyroid carcinoma, using ultrastructural methods and cytochemical techniques for peroxidase and acid phosphatase activities. The most striking ultrastructural features were: disappearance of secretory granules or apical vesicles; nuclei with an irregular shape containing stage I and II inclusions; two types of cells: with dilate endoplasmic sacks filled with a granular material, or rich in closely packed mitochondria, some of them showing dystrophic lesions with round, dense bodies or tightly packed parallel filaments and numerous phagolysosomes. The peroxidase activity was present as black DAB precipitates in the nuclear envelope and sometimes around colloid droplets, or was completely absent from some cells. The acid phosphatase activity was found as unhomogeneous Pb precipitates inside the lysosomes. The ultrastructural and cytoenzymological pattern suggests marked alteration of the synthesis, storage and secretion of the thyroid hormones. Keeping in mind that results of radio-iodine treatment depend mainly on the histologic type of the thyroid carcinoma, the cooperation between radiotherapeutist and pathologist is emphasized. PMID- 6270780 TI - Demonstration of a melanocyte-stimulating action of a pineal peptide extract. PMID- 6270779 TI - [Effect of cyproterone acetate on the secretion ACTH, cortisol and testosterone, and on the urinary elimination of 17-OHCS and 17-CS in hirsute females]. PMID- 6270781 TI - [Salivary gland cylindromas. Retrospective study of 65 cases (author's transl)]. AB - A retrospective study of prognostic, clinical, and histological features in 65 cases of salivary gland cylindromas treated in the Hopital Salpetriere, Paris, was conducted. Vital prognosis was related to the initial size of the tumour, the presence of pain, and evidence of cellular anomalies or invasion of neighbouring structures on histological examination. Metastatic spread is related to peritumoural extension, and local recurrence has a prognostic value because of its relationship to later metastatic involvement. PMID- 6270782 TI - Familial pyrophosphate arthropathy. Occurrence and Crystal Identification. AB - Hereditary pyrophosphate arthropathy has been observed in three Swedish families and in a few other caucasian populations. The inheritance is most probably autosomal dominant with a variable penetrance. The most severe cases have been found in homozygotes among isolates of immigrants in Slovakia and Chile. Studies on genetic and etio-pathogenetic factors in hereditary pyrophosphate arthropathy, and the utilization of new diagnostic techniques for crystal identification, are important approaches towards a further understanding of the disease. PMID- 6270783 TI - Measurement of soluble pyrophosphate in plasma and synovial fluid of patients with various rheumatic diseases. AB - Soluble pyrophosphate was measured in the plasma and synovial fluid of various groups of patients and in the plasma of two control groups. The two control groups consisted of 13 healthy subjects and 19 patients suffering from benign lumbar back pain. The other group of patients had rheumatoid arthritis (RA) (14 plasma and 19 synovial fluid examinations), osteoarthrosis (OA) (19 plasma and 26 synovial fluids) and articular chondrocalcinosis (ACC) (27 plasma and 43 synovial fluids). The level of soluble pyrophosphate in the plasma was 3.5 mumol/l in healthy subjects, 4.0 mumol/l in patients with lumbar back pain, 4.1 mumol/l in individuals having OA and 3.5 mumol/l in the group suffering from RA as well as for those with ACC. The differences between these values are not significant statistically. In the synovial fluid the values were 4.6 mumol/l for the group with RA, 12.7 mumol/l for those with OA and 34.2 mumol/l in the group having ACC. If a normal distribution of these values is assumed and the average values and standard deviations recalculated for each group after elimination of cases more than 3 standard deviations above the mean, then we obtain 9.8 mumol/l for the group with OA and 23.8 mumol/l for those with ACC. The difference between the group with RA and that with OA is highly significant (p greater than 0.0001). Even more significant is the difference between the group with RA and ACC (p less than 0.0005). The difference between the OA and the ACC is also highly significant (p less than 0.001). On the basis of these observations various mechanisms leading to the pyrophosphage crystal deposition disease are discussed. PMID- 6270784 TI - Evaluation of quartz crystals in rat lungs by X-ray diffractometry. AB - The NIOSH (National Institute of Occupational Safety and Health)-recommended procedure of determining quartz through X-ray diffractometry was applied to rat lungs. Using a compressed air nebulizer inside a closed chamber, 150 male Sprague Dawley rats were exposed to monodisperse particles of quartz for varying time intervals. The rats were subsequently sacrifice by cervical dislocation and their lungs were removed for ashing inside a muffle furnace at 600 degrees C for 4 hr. The ashed materials were then filtered onto silver membranes and scanned by an / ray diffractometer. The results, expressed in MG SiO2 were compared to standard values derived from PVC filters. The values suggested that the NIOSH method could be used by biochemists to quantify minerals and crystalline structures in organic materials. PMID- 6270785 TI - Basic mechanisms of 'calcium antagonists' (slow channel blockers). PMID- 6270786 TI - Electrophysiology and antiarrhythmic effects of calcium antagonists. PMID- 6270787 TI - Calcium antagonists (calcium channel blockers). PMID- 6270788 TI - Multiple opiate receptors: alcohol selectively inhibits binding to delta receptors. AB - The addition of ethanol or other aliphatic alcohols to rat brain membranes strongly inhibits binding of enkephalins at concentrations at which little inhibition of opiate alkaloids is seen. Inhibition is reversible, and potency increases with chain length of the alcohol. The results suggest that delta receptors are considerably more sensitive to alcohols than mu receptors. This is the first demonstration of selective inhibition of one of the postulated classes of opiate receptors by a reagent that is not a ligand for the receptor. PMID- 6270789 TI - Fc and C3b receptors on pulmonary endothelial cells: induction by injury. AB - Receptors for the activated third component of complement and for the Fc portion of immunoglobulin G are not expressed by apparently normal bovine pulmonary endothelial cells, but are expressed when the cells are exposed to white cell lysates or are infected with influenza or cytomegalovirus. The unmasking of these latent receptors may contribute to the pulmonary inflammatory response characteristic of, for example, anaphylaxis and to those lung diseases characterized by the deposition of immune complexes. PMID- 6270790 TI - Differentiation of respiratory and abortigenic isolates of equine herpesvirus 1 by restriction endonucleases. AB - Viruses classified by immunologic criteria as equine herpesvirus 1 cause respiratory disease and abortion in horses. Restriction endonuclease analyses of the DNA's of viruses from animals with respiratory disease and from aborted fetuses show that the patterns for respiratory viruses, while similar to each other, are entirely different from the patterns for fetal viruses. It is therefore proposed that the DNA restriction endonuclease patterns of fetal and respiratory viruses analyzed in this study be designated as prototypic of equine herpesvirus 1 and 4, respectively. PMID- 6270791 TI - Brain aging correlates: retardation by hormonal-pharmacological treatments. AB - Mid-aged rats were either adrenalectomized and chronically maintained, or left intact and treated daily for a 9- to 10-month period with a potent analog of the peptide adrenocorticotropin (residues 4 to 9), which has some stimulant properties, or with the neural stimulant pentylenetetrazole. All three treatments reduced hippocampal morphologic correlates of brain aging (neuronal loss, glial reactivity). The pentylenetetrazole and peptide treatments also improved reversal learning. These results suggest that certain endogenous peptides, with stimulant properties, may also exert long-term, trophic effects on brain structure and function. PMID- 6270792 TI - Hypothyroidism elicits electrophysiological noradrenergic subsensitivity in rat cerebellum. AB - discharge rats of Purkinje neurons were compared in control and hypothyroid adult rats. Purkinje neurons in hypothyroid rats fired significantly faster and were less sensitive to iontophoretically applied norepinephrine than those in control rats. The subsensitivity of the Purkinje neurons appeared to be primarily due to an alteration in the beta-receptor--adenylate cyclase complex, because the sensitivity of these cells to locally applied N6-monobutyryl adenosine 3'-5' monophosphate (N6 cyclic AMP) did not change significantly. The sensitivity of the Purkinje neurons to norepinephrine could be restored in hypothyroid rats by administration of triiodothyronine. PMID- 6270793 TI - Increased pyrophosphate in fibroblasts and lymphoblasts from patients with hereditary diffuse articular chondrocalcinosis. AB - The metabolic and genetic factors leading to deposition of calcium pyrophosphate crystals in cartilage of patients with chondrocalcinosis are not well understood. Analysis of cultured fibroblasts and lymphoblasts from 12 affected members of a large kindred showed a mean concentration of intracellular inorganic pyrophosphate two times greater than that in cells from unaffected family members or normal, unrelated volunteers. Increased intracellular pyrophosphate may, therefore, be a biochemical marker for the heterozygous expression of the chondrocalcinosis gene. PMID- 6270794 TI - [Hairy cell leukemia. I.--Clinical, biological and evolutive study on twenty eight cases (author's transl)]. AB - A clinical and laboratory evaluation of 28 patients with hairy cells leukemia is performed. Twenty-two had splenomegaly and all but one had a pancytopenia with 5 to 70% of hairy cells in blood. A tartrate-resistant acid phosphatase activity was positive in the hairy cells of 11 patients of 14 studied. In all patients a myelofibrosis and a leukemic infiltration were found in a bone-marrow biopsy of iliac crest. Hemodilution by splenomegaly, mild hemolysis and dyshematopoiesis were observed in 10 patients by a 51Cr or 59Fe isotopic exploration. In seven cases an immunological study of the hairy cells was performed, a high percentage of the leukemic cells of these 7 patients had polyclonal surface Ig but without resynthesis of monoclonal S Ig which is a feature usually associated with B lymphocytes. In the blood of these patients normal T and B lymphocytes were decreased. A splenectomy was done in 12 patients (43%) always for severe pancytopenia Splenectomy was not randomised. Spleen weights ranged from 1 085 to 3 600 g. In splenectomised patients the level of hemoglobin, segmented cells and thrombocytes was significantly higher after surgery. The survival rate is better in the splenectomised group (median survival 57 months) than in the non splenectomised group (median survival 19 months). Infectious diseases were frequent in all patients but less after splenectomy. Fourteen patients died, 8 owing to pancytopenia. PMID- 6270795 TI - [Hairy cell leukemia. II.--Critical review of literature. Part. 1:diagnosis and evolution (author's transl)]. AB - Hairy cell leukemia (in french: leucemie a tricholeucocytes) is a distinctive form of chronic leukemia with splenomegaly, pancytopenia, presence of a particular circulating mononuclear cell, myelofibrosis and leukemia infiltration of the bone marrow. Generally, a tartrate resistant acid phosphatase activity is found in the leukemic cells. Hairy cell leukemia is the main leukemia in which the treatment is first surgical. Splenectomy has a good result for survival median and infectious risk. The disease is still mysterious for the origin of the leukemic cell; monocyte, B lymphocyte or in few cases T lymphocyte, even though a spectrum of immunological patterns pleads for the B lymphocyte origin of the leukemic cell. PMID- 6270796 TI - [Hairy cell leukemia. II.--Critical review of literature. Part 2: the hairy cell origin (author's transl)]. AB - Hairy cell leukemia (in french: leucemie a tricholeucocytes) is a distinctive form of chronic leukemia with splenomegaly, pancytopenia, presence of a particular circulating mononuclear cell, myelofibrosis and leukemic infiltration of the bone marrow. Generally, a tartrate resistant acid phosphatase activity is found in the leukemic cells. Hairy cell leukemia is the main leukemia in which the treatment is first surgical. Splenectomy has a good result for survival median and infectious risk. The disease is still mysterious for the origin of the leukemic cell; monocyte, B lymphocyte or in few cases T lymphocyte, even though a spectrum of immunological patterns pleads for the B lymphocyte origin of the leukemic cells. PMID- 6270797 TI - [Prognosis of adult non-Hodgkin's lymphomas. Retrospective study of sixty-six patients (author's transl)]. AB - The authors studied the prognosis of 66 adults NHL according to: histological, clinical, biological and evolution criteria. The patients were treated by polychemotherapy or polychemo-and radiotherapy. Three years actuarial survival compared by the X2 test permits to consider only two independent prognostic factors: histologic type and complete remission due to treatment. PMID- 6270799 TI - [A study of the effects of procyanidol oligomers on capillary resistance in hypertension and in certain nephropathies (author's transl)]. AB - The effects of procyanidol oligomers on capillary resistance disorders in hypertensive and diabetic patients were studied. The drug was administered in a daily dose of 150 mg. Concordant results were obtained in two investigations as follows: during an open trial in 28 patients, capillary resistance (measured by the technique of Lavollay) rose from 15,4 +/- 1,8 cm Hg to 18,1 +/- 3,2 cm Hg (p less than 0,0005); and during a double blind trial versus placebo (25 patients) capillary resistance rose from 14,6 +/- 0,98 cm Hg to 18 +/- 3,35 cm Hg (p less than 0,005) in the treated group, while no significant variation was observed in the placebo group (15,5 +/- 1,30 cm Hg versus 14,7 +/- 1,3 cm Hg initially). The drug was well tolerated. PMID- 6270798 TI - [Congenital virilizing adrenal hyperplasia due to 11 beta-hydroxylase deficiency. Study of eleven cases (author's transl)]. AB - Eleven cases of congenital virilizing adrenal hyperplasia are studied. This study leads to point out some peculiar characteristics: virilization of external genitalia in girls is most usually important; arterial hypertension is delayed, usually after some years; plasma androgens and desoxycortisol are highly elevated; plasma 17-hydroxyprogesterone is slightly elevated and this may be misleading; good results of treatment which must preferably use hydrocortisone since plasma cortisol is sometimes low; long-term risks in treated children include chronic hypertension in both sexes, advanced puberty in girls and pubertal gynecomastia in boys. PMID- 6270800 TI - [Montgomery xanthomatosis (author's transl)]. AB - A typical case of Montgomery xanthomatosis is reported by the authors. Out of this case and of a hundred other ones gathered from the literature, they outline the main features of the disease. It is revealed as being stereotyped and particularly the prognosis is usually good. Whether the ultra structural study may show Langerhans corpuscules or not, it brings us a material to the discussing of border-line cases (xanthomatosis with diabetes insipidus, Hand-Schuller Christian disease with cutaneous lesions similar to Montgomery's. PMID- 6270801 TI - [Hyperuricosuria and urinary lithiasis in paraplegic patients (author's transl)]. AB - One hundred and thirteen urinary stones fron spinal cord injured patients contain urates in 65,5% of cases. In a prospective study of 21 paraplegic patients, an early increase of uricosuria was found as well as a late and moderate elevation of uricemia. The origin of these changes of the metabolism of uric acid is discussed. Therapeutic consequences from these results are studied. PMID- 6270802 TI - [Mycotic aortic aneurysms (author's transl)]. AB - Mycotic aortic aneurysms were detected in four men aged between 47 and 73 years. The infective nature of the lesion was obvious in one case, while in the other three cases the diagnosis was suspected after radiological examination and confirmed by pathological and bacteriological investigations of the aneurysm wall. Radiological signs which suggest the presence of an infective aneurysm include rupture of the aneurysm, a rapidly progressing course, and unusual locations or morphology. Diagnosis is important as therapy differs from that of other types of aneurysm. PMID- 6270803 TI - [Headache affecting the pigmented zones of Ota's naevus: clinical study in four cases (author's transl)]. AB - Four women, aged between 23 and 69 years, who had developed Ota's naevi either during the menarche (3 cases) or the first pregnancy (1 case), complained of headache in the frontal and orbital zones affected by the naevi. These headaches bore no relationship to those normally described as occurring in this region. They increased in severity, at the same time as the intensity of the pigmentation of the naevus, in each patient during the menstrual periods, and did not respond to conventional analgesics. PMID- 6270804 TI - [Appraisal of oncological and cosmetic results of the surgical treatment of 138 baso-cell carcinomas (author's transl)]. AB - The series concerning the treatment of baso-cell carcinoma often present large numbers, but the treatment has often been made according to different modalities, and several operators of varying experience have been involved. Also the cosmetic result has never been evaluated, which is strange since it is the face which is mostly concerned (96%). We are presenting only 138 reviewed lesions, but operated by one operator according to the same technique: surgical excision, immediate repair, frozen section being exceptional. The local flaps were favored, the distant flaps and full thickness grafts being made only when no local flap was available. The size and the site of the lesions (mainly centrofacial) is the same as in other series. The recurrence ratio with 3 years minimum of follow-up is 2,1%. The cosmetic result was evaluated by the patients and the non-operative author. The excellent results (no visible scar, no deformity) amounted to 61,4%, the good ones to 31,4% and are due essentially to local flaps. These results are discussed. PMID- 6270805 TI - [Association between mycosis fungoides and myasthenia gravis (author's transl)]. AB - Myasthenia gravis characterized by physical examination, electrophysiologic abnormalities, ephedronium test, without thymoma, associated in a 47 year-old man, with mycosis fungoides. Myasthenia gravis began two years after the onset of mycosis fungoides, and the patient died of infection four years after the development of the cutaneous lymphoma. At autopsy, there was no visceral involvement. Myasthenia gravis and lymphoma rarely coexist; the occurrence of myasthenia gravis shortly after the onset of mycosis fungoides suggests that the two disorders may be linked by immunological abnormalities. PMID- 6270806 TI - [Serial infections with two herpesviruses after renal transplantation: virological and ultrastructural features (author's transl)]. AB - A fatal case of active viral infection in a transplant patient is reported. On the second month post transplantation, Cytomegalovirus (CMV) infection occurred. It was attested by clinical and radiological symptoms, transient increase of transaminases and seroconversion with CMV antigen. Patient apparently recovered, but on the third month mucocutaneous Herpesvirus hominis (HVH) infection started. It was rapidly complicated by hepatitis and encephalitis which are directly responsible for the fatal outcome. A strain of HVH type 1 was easily isolated from oral and genital lesions. Immunofluorescence with HVH antiserum was positive on post mortem sample of liver biopsy Ultrastructural changes and intranuclear Herpesvirus like particles were demonstrated in liver. Etiology of hepatitis and management of immunosuppressive therapy are discussed. PMID- 6270807 TI - [Primary syphilitic proctitis associated with liver involvement (author's transl)]. AB - A primary syphilitic proctitis histologically simulating a Crohn's disease, has been found in a 34 year-old-man, who is married and father of three children. He presents also a biological and histological liver involvement. He completely recovered wih antibiotics. Review of the literature is done. Syphilitic serology studies and treponems research in rectal discharges should be done in case of anorectal lesions, histologically benign, but of uncertain etiology, if the patients is known as homosexual or not. PMID- 6270808 TI - [Non lymphoid cells of the peripheral lymphoid organs. Recent morphological progress and current understanding (author's transl)]. AB - Peripheral lymphoid organs have two groups of non lymphoid cells: that of macrophages which belongs to R. van Furth's system of mononuclear phagocytes and the one made of elements for long labelled "reticular cells". The use of electron microscopy has permitted a good understanding of these elements and a recognition of ubiquitous cells (fibroblasts, endothelial cells) and specific cells to the lymph node which seem to play a very important role in the immune reactions (interdigitated cells, reticulum cells, dendritic cells). We describe their light and electron microscopical features and histoenzymatic characters, and present the actual concept of their origin and function. The latest discoveries on their classification are reviewed and have permitted to distinguish three categories of non lymphoid cells: first the group of reticulum cells (reticulum cells and dendritic cells), second that of mononuclear phagocytes (macrophages and interdigitated cells) and third that of endothelial cells. Other elements of questionable existence or insufficiently known do not appear in this classification: dark cells, Steinman and Cohn's dendritic cells and cells containing Birbeck's granules. PMID- 6270809 TI - [Haemodynamic and metabolic effects of exercise test in diabetic patients with arteritis treated or not with nicergoline (author's transl)]. AB - A randomised crossover study in eleven diabetic patients with arteritis compared the effects of nicergoline (2,5 mg i.v.) or placebo on haemodynamic and metabolic parameters after exercise tests. Haemodynamic modifications after effort following placebo administration were typical: raised systolic blood pressure, and increased heart rate and myocardial oxygen requirements (systolic BP x heart rate). Modifications after similar effort following nicergoline involved an increase in systolic B.P. only, heart rate and myocardial oxygen requirements remaining unchanged. Blood lactic acid levels after effort and treatment were significantly higher (p. less than 0.01) than after effort without treatment. Overall metabolic and haemodynamic results demonstrate an increase in effort tolerance in diabetic patients with arteritis after nicergoline, this having been previously observed in healthy subjects. PMID- 6270811 TI - [The progeny of women with treated congenital hypothyroidism (author's transl)]. AB - The progeny of 11 women with treated congenital hypothyroidism was studied. They had 17 pregnancies that led to 2 spontaneous abortions, 2 children dead in the first month after birth, and 13 living children. These 13 children have a normal psychomotor development. One of them has a lingual ectopic thyroid as her mother, one has a congenital cardiac defect, one has a minor anomaly of toes. The authors add a previously reported familial case of hypothyroidism in a mother and her four children. PMID- 6270810 TI - [Aromatization of androgens (author's transl)]. AB - Aromatization of androgens into estrogens is an early phenomenon during phylogenesis and ontogenesis which occurs in most tissues. The mechanism which is responsible for this activity and for it's competition with 5 alpha reduction includes hypothalamo-pituitary receptors, genetic induction of sexual differentiation, binding proteins, and sulfo and glyco-conjugation. Aromatization is determinant before, during, and after ovulation, as well as in developing pregnancy. In the testis, in spite of it's importance, it's role is unknown. It's action is essential in the sexual differentiation of the brain. Aromatization in the muscle and the adipose tissue is not negligible under normal and pathological conditions. PMID- 6270812 TI - [Treatment of primary hypothyroidism in adults : dosage schedule and biological surveillance (author's transl)]. AB - Hypothyroidism in a disease with a poor prognosis, especially as the disorder is often unrecognised, or is treated empirically and without adequate supervision. Modern biological tests can establish the diagnosis, even in subclinical cases, and can also ensure close supervision of dosage to avoid long-term adverse effects of too low or too high dosage regimens. The authors prefer dl thyroxine (dl T4) to other currently employed products : wide variations in triiodothyronine (T3) activity in thyroid extracts, short abrupt action of T3, frequent intolerance to combined T4/T3. The dl T4 (or T4 at half the dosage), a prohormone active after transformation into T3 in the organism, appears to be much more convenient in use. Dosage, usually overestimated, should be an average of 165-200 microgram of 1 T4 (5 to 6 drops of dl T4). It should be progressively reduced as a function of age (approximately 16 micrograms per decade), falling to 66 microgram/day (2 drops) over 80 years of age. Dosage must be temporarily reduced by 50 p. cent or more during acute affections. When first instituting treatment it is imperative to administer the product in stages - of two weeks in the young and of one month in elderly or debilitated patients - because of the long half-life (7 days) of T4. Initial dosage should be half the predicted final dosage in the young, and even less in the elderly. If these rules are respected, the association of beta-blocking agents, in particular, becomes of less and less value. Supervision by repeated biological tests is essential, clinical investigations detecting subclinical over - or under - compensated forms with difficulty. Modern biology has completely changed the classical rules, and often permits reduction of dosage and subsequent suppression of many adverse reactions. Equilibrium is reached when TSH levels become normal and T3 levels within normal limits. This often leads to T4 levels slightly higher than normal, as the concentrations of T3, the only active compound, are derived from plasma T4 and not, as in normal subjects, from the plasma T4 (70 p. cent) and the thyroid (30 p. cent). The TRH test, to detect markedly subclinical biological forms of hypo - or hyper - thyroidism, is only rarely necessary. PMID- 6270813 TI - [Puberty and adult height in congenital virilizing adrenal hyperplasia (author's transl)]. AB - A study of 52 pubertal patients having been treated for virilizing adrenal hyperplasia (38 females and 14 males) leads to emphasize the importance of the precocity and accuracy of treatment. In patients with delayed treatment, mainly when they are girls, an advanced or true precocious puberty will increase the advance of bone age and reduce adult height. But, whatever be the time of onset of treatment and of pubertal achievement, no important or sustained sexual improvement was found in patients receiving a well-controlled treatment. PMID- 6270814 TI - [Thyroiditis : diagnostic and therapeutic features (excluding Graves disease and myxedema) : 13 case-reports (author's transl)]. AB - The incidence of De Quervain's thyroiditis is probably underestimated, since a radioiodine uptake study is not performed in all cases. An isolated cold nodule can reveal Hashimoto's thyroiditis. In other cases, the diagnosis is suggested by otalgia and positive tests for antimicrosomial antibodies. Surgery is often necessary because of the possible association with cancer ; the procedure should be extensive since spontaneously the outcome is often thyroid insufficiency. PMID- 6270815 TI - [Hypophosphatemia in total parenteral nutrition (author's transl)]. AB - There is a hypophosphoremic syndrome first discovered without parenteral nutrition (TPN) then on patients with TPN. It appears preferentially in patients suffering from denutrition and receiving an important caloric intake, particularly with glucose. Clinical picture associates psychic, neurologic and respiratory disorders. Phosphatemia is often below 10 mg/l or 0,33 mmol/l, with decrease or even disappearance of phosphatemia. The mechanism is still baby known. During TPN, it occurs an intra-cellular passage of phosphate whose consequences are anomalies in red blood cell metabolism, with decrease of tissular oxygenation, in white cell function, platelet function, central nervous system, muscle, liver and acid-base equilibrium. Treatment must be first preventive with careful control of phosphatemia and systematic intake of phosphates. Curative treatment associates correction of hypophosphatemia and simultaneous decrease of caloric intake. PMID- 6270816 TI - [Non-invasive hemodynamic evaluation of male impotence (author's transl)]. AB - Since male impotence may be caused by a variety or non vascular factors, more precise techniques for identifying potential operative candidates must be developed. Our protocol includes nocturnal penile plethysmography, Doppler velocity at 8 levels, penile systolic pressure, electromagnetic blood flow measurement, penile reactive hyperemia. This non-invasive protocol applied on 21 patients aims to select those who would benefit from direct penile revascularization procedures. We are studying the venous emptying rate for more complete physiologic approach. PMID- 6270817 TI - [Tiapride and rehabilitation (author's transl)]. AB - Common obstacles to rehabilitation of patients having suffered severe trauma are lability of attention and relational difficulties with the physician. Major psychopharmaceutic agents often hinder adequate motor function. Tiapride was used in twelve patients with psychological and/or motor difficulties slowing rehabilitation. The average daily dosage was 600 mg orally (divided in three doses of two tablets each), during seven weeks. In three patients, the difficulties disappeared completely. The major problems disappeared in six patients and were improved in one. The treatment failed in two cases. Tolerance was excellent in spite of the poor condition of many of the patients. The only side-effects were marked drowsiness in one patient and negligible drowsiness or insomnia in four. PMID- 6270818 TI - [Effects of trichlorethylene and its metabolites on the rate of sister chromatid exchange. In vivo and in vitro study on the human lymphocytes (author's transl)]. PMID- 6270819 TI - [Synthetic progestogens (author's transl)]. AB - Synthetic progestogens are divided into three major groups of steroids which derive from the basic steroid nucleus. Several routes of administration can be used. The characteristics (progestomimetic, progestational, and hormonal) of the different progestogens are variable. The different compounds are therefore of therapeutic value in specific indications. For some of these a study of the literature was carried out. We studied : anomalies of the menstrual cycle (involving chronology, menstruation...); the premenstrual syndrome with or without benign mastopathy ; menopause (premenopause and established menopause). The differences in the tolerance of the different compounds in use are recalled, in connexion with contra-indications and major and minor side-effects. The wide differences between the various compounds can make the choice of the less than right progestogen greater than difficult. PMID- 6270820 TI - [Effects of progestogens on the metabolism of lipids and carbohydrates. Practical consequences (author's transl)]. AB - Estrogens which are one of the components of contraceptive less than pills greater than are incriminated in many cardiovascular accidents. These occur as a result of metabolic disorders (involving lipids and carbohydrates), of modifications in coagulation factors, etc. The possible influence of progestogens was ignored for a long time. The widespread use of these compounds, prescribed for contraception as well as during hormonal replacement therapy for absolute or relative luteinic insufficiency, makes careful monitoring of lipid and carbohydrate metabolism imperative. This position is strengthened by a preliminary review of the literature which leads to several conclusions concerning lipid and carbohydrate metabolism. PMID- 6270821 TI - [Insulin allergy (author's transl)]. AB - Severe allergies to insulin are uncommon. In most cases, allergy is merely a mild and transient reaction at the site of injection. Immediate reactions (generalized or localized) originate in an overproduction of specific IgE anti-insulin antibodies. Local delayed reactions are mainly due to substances other than insulin (adventitious contaminants, foreign proteins, zinc) and fit Gell and Coombs' description of type IV allergic reactions. Diathesis and the history of previous discontinued insulin therapy are important to consider. Management differs according to the severity of allergic reactions. Methods range from simple observation to desensitization. Monocomponent pork insulin is useful in a number of cases. Human synthetic insulin may possibly be responsible for allergies similar to those which occur with monocomponent insulin. PMID- 6270822 TI - [Two observations of Loa loa filariasis. Value of the diethylcarbamazine test (author's transl)]. PMID- 6270823 TI - [A clinical trial of a new class IV corticosteroid for topical use. Fifty-two case-reports (author's transl)]. AB - A study of the activity of a new corticosteroid for topical use, betamethasone in a propylene glycol vehicle, is reported. The trial was open and included 52 cases. These were mainly severe and chronic cases of atopic dermatitis, psoriasis and intractable prurigos resulting from various causes. The overall results were quite satisfactory, with 80% of good or very good responses. Itching was the feature most frequently improved by this treatment. If atopic dermatitis and psoriasis reacted well the response was particularly satisfactory in lichen planus. Evidence indicating an important effect on adrenal gland function is pointed out. This effect was demonstrated either by decreased serum cortisol levels or by decreased urinary excretion of steroids. This point should be carefully monitored, especially when treating patients with extensive skin disease. Therapy should be discontinued very progressively. PMID- 6270824 TI - [Rare abdominal and peritoneal localizations of hydatid cysts. Report on two cases (author's transl)]. AB - Rarely observed isolated peritoneal hydatid cysts, without initial hepatic lesions or later development in other viscera, were detected in two patients. The diagnosis of hydatidosis confined to the mesocolon was established pre operatively in the first case, enabling mebendazole treatment to be instituted. In the second case, however, pre-operative diagnosis was not possible, demonstrating the sometimes misleading appearances found in this affection. Aetiopathogenic and diagnostic features are briefly discussed. Therapy raises problems, and though good results have recently been obtained with benzimidazole derivatives, surgical excision in currently the only curative treatment available. Medical treatment (with benzimidazole derivatives) is indicated only in cases with multiple cysts inaccessible to surgery, or a complementary therapy to facilitate excision, reduce risks of propagation during operation, and prevent recurrences. PMID- 6270825 TI - [Current advances in cancer immunotherapy (author's transl)]. AB - Immunotherapy is still considered as an important therapeutic method in oncology. Experimental basis do exist which support the individualization of several distinct conditions of immunomodulation, some of them being associated with a therapeutic benefit. Numerous immunomodulation molecules or derivatives are now available for clinical investigations. However, effects of immunomodulators on the immune systems which depend on several unknown factors are actually not always fairly correlated with antitumoral effects. Clinical trials are therefore needed in order to demonstrate that immunotherapy remains a valuable antitumoral treatment. The authors review here the available data obtained from recent clinical studies in man. They attempt to establish the limits of immunotherapy as well as its future possibilities. PMID- 6270826 TI - [Management of hypo-uricemia (author's transl)]. AB - Hypo-uricemia (under 120 mumol/L) is not uncommon (1% of the population). Two main mechanisms are responsible (abnormal synthesis and anomalies in renal excretion), but there are a great number of possible etiologies. Except for states of stress or distress (30%), etiology focuses on drugs (60%), mainly aspirin. Malignant disease, particularly myeloma and broncho-pulmonary carcinoma which can be revealed by hypo-uricemia, are much less frequent. Genetically determined cases are uncommon. The diagnostic approach for finding the etiology should be physiopathological and focused on a simple test : uric acid clearance. PMID- 6270827 TI - Electron microscopy in the diagnosis of the bone marrow disorders of the erythroid series. PMID- 6270828 TI - [Kidney tumors in children]. PMID- 6270829 TI - [Kidney neoplasms in the adult]. PMID- 6270830 TI - [Extraction of drugs from biological material using kieselguhr]. PMID- 6270831 TI - [Role of the cytomegalovirus in the pathology of pregnancy, of the fetus and of newborn infants]. PMID- 6270832 TI - [Treatment of colonic diverticulosis and irritable colon with wheat bran]. PMID- 6270833 TI - [Treatment of catatonic states with adrenocorticotropic hormone]. PMID- 6270834 TI - Treatment of uncomplicated gonorrhea with cefotaxime. AB - Patients with uncomplicated gonorrhea due to beta-lactamase-negative Neisseria gonorrhoeae were treated with cefotaxime (1.0 g given im in a single dose) or with aqueous procaine penicillin G (APPG; 4.8 x 10(6) units given im) plus probenecid (1.0 g given orally). Genital or rectal gonococcal infection was cured in 51 (93%) of 55 patients given cefotaxime and in 23 (96%) of 24 patients given APPG plus probenecid. Gonococcal isolates from homosexual men were less susceptible to cefotaxime (geometric mean MIC, 0.021 microgram/ml) than were strains isolated from heterosexual men (geometric mean MIC, 0.012 microgram/ml; P less than 0.05). Genital infection with Chlamydia trachomatis persisted in four of eight patients given cefotaxime and was first detected after treatment in three others. Of 23 men with gonococcal urethritis who were treated with cefotaxime and followed for 11--30 days, ten (43%) developed postgonococcal urethritis; five of these were associated with chlamydial infection. Administration of cefotaxime or APPG caused equal pain, but cefotaxime was better tolerated because of the need for only one injection. Cefotaxime and APPG plus probenecid are comparable for treatment of uncomplicated genital and rectal infection with beta-lactamase-negative N. gonorrhoeae. PMID- 6270836 TI - Insulin, glucagon and glucose in the regeneration response of the liver. AB - The effects of intraduodenal glucose load on the hepatic uptake of insulin, glucagon and glucose after a 70 per cent hepatectomy were studies in anesthetized dogs. Dogs without a hepatectomy served as the control study. Data were derived from plasma concentrations in the portal vein, aorta and hepatic vein with simultaneous portal vein and hepatic artery plasma flow measurements. The concentrations of glucose in the arterial blood of dogs after a hepatectomy were less than those for the controls throughout the study, while insulin and glucagon concentrations showed no differences between the groups. Hepatic uptakes of insulin, glucagon and glucose per gram of liver perfused were significantly greater in the hepatectomy group and occurred because the amounts of these substances reaching the liver remnant were the same as those for the controls. The increased uptake of insulin and glucagon after partial hepatectomy may reflect increased binding of these hormones to the liver cell receptors, by which hepatic regeneration is induced. Increased glucose uptake could serve as the substrate for the accelerated anabolic processes. PMID- 6270835 TI - Requirement of the human chromosome 11 long arm for replication of herpes simplex virus type 1 in nonpermissive Chinese hamster x human diploid fibroblast hybrids. AB - Somatic cell hybrids between Chinese hamster (CH) lung cells (V79/380-6), nonpermissive for productive infection by herpes simplex type 1 (HSV-1), and permissive human diploid cells support productive HSV-1 infection as long as they retain human chromosome 11. Human chromosome 3 has been reported to complement nonpermissivity in (CH) Don cells (1). Intraspecies hybrids between Don/a3 and V79/380-6 cells, however, did not support HSV-1 replication, indicating lack of complementation. The block in both nonpermissive CH cell lines was determined to involve a step beyond replication of the parental viral DNA. In cell hybrids between nonpermissive Don/a23 cells and human fibroblasts containing a t(11;15) (p11;p12) translocation, HSV-1 production was dependent solely on the presence of either human chromosome 11 or the der(11) (p11 leads to qter) translocation product containing the long arm of chromosome 11. Chromosome 3 was excluded by a discordancy rate of 59%. We conclude that the long arm of human chromosome 11 carries one or more genes coding for host functions necessary for the production of progeny HSV-1 DNA. PMID- 6270837 TI - [Scintigraphy with 99mTc-pyrophosphate in patients with scapulohumeral periarthritis]. PMID- 6270838 TI - Recovery of plasma corticotrophin and cortisol levels after three-week course of prednisolone. AB - Patients with chronic airflow obstruction were given a three-week course of prednisolone 40 mg per day. The basal plasma cortisol level and response to tetracosactrin were depressed after such a course. Basal plasma cortisol and corticotrophin (ACTH) levels were measured on five consecutive days after three weeks of treatment with prednisolone and were found to rise simultaneously to control levels within three days. Pituitary and adrenal functions were depressed for four days after short high dose courses of corticosteroids and patients may be at risk if they encounter stress during this time. PMID- 6270839 TI - Intrathoracic chemodectoma with noradrenaline secretion. PMID- 6270840 TI - [Hemodynamic and coronary changes induced by various slow-channel inhibitors in chronic coronary insufficiency: nifedipine, verapamil and bepridil]. PMID- 6270841 TI - [Methodology and results of a post-marketing drug study in diarrhoeal disease of children (author's transl)]. PMID- 6270842 TI - Heparin opposes prostanoid and non-prostanoid platelet inhibitors by direct enhancement of aggregation. PMID- 6270843 TI - Factors influencing shear-induced platelet alterations: platelet lysis is independent of platelet aggregation and release. PMID- 6270844 TI - [Field experience using an inactivated vaccine in Aujeszky's disease vaccination of cattle (author's transl)]. AB - In the autumn of 1979, approximately, 400 animals on twenty farms were inoculated twice against Aujeszky's disease using an inactivated vaccine in a practice in the Achterhoek (province of Gelderland). The animals vaccinated were mainly yearlings and heifers on farms on which cattle and pigs could not be effectively separated. Adverse reactions to vaccination, other than a slight to moderate increase in body temperature, were not observed. Outbreaks did not occur in vaccinated cattle during the 1979-1980 winter season; on the other hand, Aujeszky's disease was observed in the pigs on a number of farms. The serological effects of vaccination were followed up in forty-one animals on sixteen farms. Very high antibody titres were attained, particularly after the second vaccination. Antibody titres continued to be recorded after twelve months in a number of the animals followed. PMID- 6270845 TI - Preference for drinking water containing dimercaptosuccinic acid by rats intoxicated with methylmercury. PMID- 6270847 TI - Particle size distribution of hydrocyanic acid in gari, a cassava-based product. AB - A reciprocal relationship was observed between the cyanide content of gari and particle size. Hydrocyanic acid (HCN) content was positively correlated (r = 0.62) with sugar content but the correlation with starch content was poor (r = 0.33). From both the nutritional and toxicological standpoints, it would appear that larger particles size in gari is beneficial. PMID- 6270846 TI - The effect of long term treatment with disulfiram on content of cytochrome P-450 and on benzo(a)pyrene monooxygenase activity in microsomes isolated from the rat small intestinal mucosa. AB - Disulfiram (DS) was administered perorally once a day to rats for 30 days to investigate the effects on cytochrome P-450 content and benzo(a)pyrene (BP) monooxygenase activity in microsomes isolated from the small intestinal mucosa. 50 mg or 100 mg DS/kg body weight caused a dose-related increase in BP monooxygenase activity, whereas the content of cytochrome P-450 was increased at the higher dose only. Similar absorption characteristics of cytochrome P-450 and turnover rates for BP on the basis o f cytochrome P-450 was observed among the different microsomal preparations. The addition of DS or diethyldithiocarbamate (DDTC) to incubates of intestinal microsomes inhibited BP monooxygenase activity. Microsomes isolated from DS-treated rats were however less sensitive to in vitro inhibition by DS. PMID- 6270848 TI - Vitamin D in plasma: quantitation by a nonequilibrium ligand binding assay. AB - The concentration of vitamin D was determined in human and bovine plasma samples under various physiological and nonphysiological conditions using a nonequilibrium ligand binding assay. Prior to ligand binding analysis the vitamin D in the plasma organic extracts was purified using chromatographic procedures involving Lipidex-5000 and high performance liquid chromatography. The use of a nonequilibrium assay system greatly increased the sensitivity of our assay allowing for a minimum volume of the initial plasma sample. The vitamin D levels in plasma responded to increased sun exposure as well as to the intoxication with vitamin D3. Analysis of a plasma sample from a vitamin D-deficient patient revealed that lipid interference was not a factor in this ligand binding assay. PMID- 6270849 TI - Steroids as potential modulators of angiotensin receptors in bovine adrenal glomerulosa and kidney. AB - To test the hypothesis that there is feedback inhibition of adrenal angiotensin receptors by substances released in response to the peptides, we measured binding of labeled angiotensins in the presence of various steroids. Approximately half of the 70 steroids tested inhibited binding of labeled angiotensin II and III to intact and broken cells from bovine adrenal glomerulosa and kidney, but the concentrations required for inhibition were relatively high. The most potent inhibitors were 3 alpha, 5 beta tetrahydroaldosterone and tetrahydrodeoxycorticosterone (ID50 = 8 x 10-5 M). Kinetic analysis showed that inhibition was mostly competitive. among steroids whose reduced congeners were tested, potency increased in the sequence: parent steroid less than 5 alpha dihydroderivative less than 5 beta dihydro derivative less than 3 alpha, 5 beta tetrahydro-derivative. Tetrahydrodeoxycorticosterone inhibited aldosteronogenesis by intact cells at concentrations that inhibited angiotensin binding. Steroids differentially inhibited binding of labeled angiotensins in II and III, and discriminated between receptors in adrenal glomerulosa and kidney. The results provide additional evidence for heterogeneity of angiotensin receptors, and lead to the prediction that any normal or pathological inhibition of angiotensin receptors by steroids will be mediated by reduced derivatives. PMID- 6270850 TI - Hormone mediated changes in monoamine stores and regulation of enzymes of biosynthesis and metabolism in the rat adrenal gland. Influence of progesterone, estradiol, ACTH and testosterone administration. AB - The influence of daily injections, for 7 days, of 17-hydroxyprogesterone caproate (8 mg/100 g body weight pro die), of estradiol-17 beta 17-undecylate (1.2 mg/100 g body weight pro die) of testosterone enanthate (2 mg/100 g body weight pro die) and of ACTH (4 IU/100 g body weight pro die) to male Wistar rats, weighing 200 g, were investigated. Monamine storage and regulation of enzymes phenylethanolamine N-methyl transferase (PNMT), monoamine oxidase (MAO) and catechol-O-methyl transferase (COMT) were studied in the adrenal glands. Results from testosterone treated rats were especially significant : the hormone administration decreased the stores of dopamine and norepinephrine (epinephrine precursors), stimulated PNMT and decreased metabolism of epinephrine by MAO and COMT. However, the adrenal content of epinephrine remained in the normal range. These findings suggest that uptake of epinephrine is accelerated in peripheral tissues by testosterone. PMID- 6270851 TI - Double antibody enzyme immunoassay of cortisol in bovine plasma. AB - A double antibody enzyme immunoassay of plasma cortisol was established using beta-galactosidase from E. coli as the tracer. Cortisol-21-hemisuccinate was conjugated with beta-galactosidase using a water-soluble carbodiimide. An antibody raised in the rabbit against cortisol-21-hemisuccinate-bovine serum albumin was used as the first antibody and anti-rabbit gamma-globulin goat serum was used as the second. The sensitivity of the present enzyme immunoassay was 25 pg/tube. The method satisfied general reliability criteria regarding specificity, accuracy and precision. Plasma cortisol levels estimated by the enzyme immunoassay was highly correlated (r = 0.99, P less than 0.005) with the levels measured by radioimmunoassay. This method of enzyme immunoassay was found to be of practical application to the routine assay of plasma cortisol. PMID- 6270852 TI - Primary cultures of Leydig cells from rat, mouse and pig: advantages of porcine cells for the study of gonadotropin regulation of Leydig cell function. AB - Primary cultures of interstitial cells were prepared from the testis of mice, rats, and pigs. The cells were grown in a defined medium supplemented with low (0.1%) serum and insulin, transferrin and epidermal growth factor. Comparisons of the interstitial cell cultures from the three species were made for plating efficiency, cell survival, maintenance of hCG receptors and maintenance of steroidogenic responsiveness to hCG. The porcine cultures had a higher plating efficiency and higher hCG receptor levels per cell than Leydig cells from either rodent. Additionally, the porcine cells showed an increase in testosterone (T) production with hCG stimulation throughout their lifespan in culture while the rodent cultures showed a decrease in T stimulation with time with no stimulation by day 6 in culture. These data indicate that species differences exist in hCG receptor concentrations per cell, the maintenance of hCG receptors and steroidogenic response in culture. The initial high survival, purity and continued functional response of porcine interstitial cell cultures make them a superior system for the study of gonadotropin regulation of Leydig cell function. PMID- 6270853 TI - Regulation of thymocyte proliferation: effects of L-alanine, adenosine and cyclic AMP in vitro. AB - Thymocyte proliferation in culture was studied by measuring the mitotic activity at hourly intervals during two periods, 0-13 and 18-30 h after the start of incubation. In particular, we investigated the effect of increased levels of cyclic AMP which are generated during preparation of the cells due to influence of endogeneously released adenosine. This cycle AMP surge, which could be blocked by combined treatment with adenosine deaminase and theophylline, caused a transient inhibition of proliferation at a stage 4 h prior to mitosis, suggesting a cyclic AMP-sensitive step in that part of the cell cycle. The temporary rise of cyclic AMP did not trigger any resting cells to proliferate. Addition of adenosine deaminase plus theophylline stimulated mitotic activity in the 18-30 h interval, possibly by blocking the influence of continuously released adenosine. The growth of thymocytes was partly synchronous, which was independent of the early cyclic AMP peak. About half of the cycling cell population was shown to be critically alanine dependent. L-alanine deprivation inhibited these cells at a stage in the cell preparation, release of prostaglandins and changes in cell viability could not explain the synchronous growth. The selective elimination of late S- and G2-phase cells during preparation of thymocytes is offered as a possible explanation instead. PMID- 6270854 TI - [Oncogen localization in the DNA composition of simian adenoviruses. II. The identification in SV20 virus DNA of a specific fragment possessing transforming activity]. AB - Simian adenovirus 20 DNA was specifically cleavered by restriction endonucleases EcoRI, BamHI, XbaI and HindIII. The transformation activity of the DNA digest was investigated. BamHI, XbaI, and HindII DNA digests were able to transform the primary rat kidney cell culture (Wistar) as well as the native SV20 DNA. The transforming activity was revealed in a specific fragment of the viral DNA, obtained after the treatment of the DNA with BamHI (fragment B), with molecular weight 5.4 x 10(6) dalton. This fragment is located in the left end of the viral genome. The lack of cell transformation by the EcoRI-hydrolysate of viral DNA may serve a proof of the extremely left position of the oncogene in the viral genome, since of EcoRI-fragment chips off a fragment with molecular weight 3 x 10(5) dalton fr om the left side of DNA molecule. PMID- 6270855 TI - [Effect of cytochrome c on the activity of lysosomal enzymes in the rat liver under hypobaric hypoxia]. AB - Under conditions of three-hour hypobaric hypoxia the total activity of acid phosphatase and DNase in the rat liver somewhat lowers. The activity of free enzymes increases by 27 and 37%, and that of bound ones decreases by 42 and 24 %, respectively. Cytochrome c being introduced to hypoxic animals, the total activity of the enzymes does not significantly change. Under these conditions the activity of free acid phosphatase increases by 16%, and bound one decreases by 24 %. The activity of free acid DNase somewhat rises (by 12%) and that of bound one lowers (by 15%). A preliminary administration of cytochrome c to the organism prevents the development of pronounced changes in the activity of the studied lysosomal enzymes in the liver under grave hypoxia. PMID- 6270856 TI - [Structural and functional features of membranes with different cholesterol content]. AB - The paper deals with the present-day ideas of the structure and functions of membranes with different cholesterol content and relative sterols. The data on the molecular bases of lipids and sterols interaction in model systems, monolayers and liposomes, are obtained using 1H-, 13C-, 31P-NMR, EPR investigations, methods of differential scanning calorimetry and X-ray diffraction. The methods for obtaining biomembranes with the different content of cholesterol and its effect on the membrane structure and functions are discussed. Reasons are analyzed of principal differences in behaviour of biomembranes with different content of cholesterol and respective model systems. The nature of cholesterol binding with phospholipids, proteins and its asymmetric distribution in biomembranes is considered. The cholesterol effect on the erythrocyte membrane charge density and possible changes in structural and functional properties of the membranes are demonstrated through the author's own experimental data. It is concluded that the cholesterol effect on structure and functions of the membranes is determined to a considerable extent by their phospholipid composition, localization and phase behaviour of phospholipids and sometimes depends on the membrane charge density. PMID- 6270857 TI - [Alkaline phosphomonohydrolases of Pacinian corpuscles]. AB - Research on quantitative determination of activity of alkaline phosphatase (EC) 3.1.3.1) and 5'-nucleotidase (EC 3.1.3.5) of Pacinian corpuscles was performed. Two enzymes are shown to be different, which are distinguished by the pH optimum and substrate concentration. Alkaline phosphatase is found to be more active in splitting glycerophosphates. The alkaline phosphatase activity towards beta glycerophosphate and alpha-naphthylphosphate is caused by various enzyme molecular forms with different electrophoretic mobility. The phosphatase and 5' nucleotidase are shown to be activated by Mg2+ and Ca2+. PMID- 6270858 TI - Ultrastructural histochemistry of infantile digital fibromatosis. AB - A case of infantile digital fibromatosis was studied by light and electron microscopy histochemistry. Using two different acidic solutions of phosphotungstic acid at varying pHs, the round inclusions characteristic of this tumor were shown to have a high protein content with little or no carbohydrates. The histochemical reactivity of the inclusions was similar to that of the cytoplasmic microfibrils in the tumors cells and consistent with the idea that both the inclusions and the microfibrils represent actin. There is, however, no definite proof that the tumor cells are myofibroblasts. At the present time, this tumor should be viewed as a peculiar expression of deranged assembly or metabolism of filamentous proteins or both. PMID- 6270859 TI - Coin lesion of the lung in a 19-year-old male. PMID- 6270860 TI - [Benign Herpes simplex virus infection in a new born infant]. PMID- 6270861 TI - [Anterior interosseous nerve syndrome. A review with 2 case histories]. PMID- 6270862 TI - [Granulated guar-gum (Slocose) for non-insulin dependent diabetic patients]. PMID- 6270863 TI - [Mucinous carcinoma of the skin]. PMID- 6270864 TI - [Pyarthron with Clostridium perfringens after injections of a steroid preparation]. PMID- 6270865 TI - [Control of the parallel release of ACTH, beta-endorphin + beta-LPH and their fragments in cultured adenohypophyseal rat cells]. PMID- 6270866 TI - [Nuclear T3 receptors in the brain and cerebellum of developing rats]. PMID- 6270867 TI - ["Oat cell" carcinoma of the lung: an ambiguous diagnosis, treatment to be reconsidered]. PMID- 6270868 TI - [Treatment experience with retrorectal cysts]. PMID- 6270869 TI - [Blood kinins in diseases of the abdominal organs]. AB - Under investigation were 151 patients with peptic ulcer (68), postresective disorders (8), non-incarcerated hernias (20), chronic gastritis (11), appendicitis (7), cholecystitis (5) and healthy people (32). The ulcerous disease, especially when complicated, is accompanied by increased activity of the kallikrein-kinin system. Certain deviations are observed in postresective disorders. These disorders were found to increase within the first day after operations on the stomach and in less degree after other interventions. Later these symptoms tended to normalize. The kinin activity was influenced by the severity of the operation trauma and anesthesia. Sufficient preoperative preparation and use of kontrikal resulted in less disturbance in the kinin system. PMID- 6270871 TI - Ultrastructure of macrophages and karyolytic bodies in small intestinal villi of macaque monkeys and baboons. AB - Macrophages in the lamina propria at the tips of small intestinal villi in 41 of 51 macaque monkeys were filled with eosinophilic, autofluorescent, periodic acid Schiff-positive globules, hematoxyphilic and Feulgen-positive granules, vacuoles, and iron and lipochrome pigments. The Feulgen-positive granules were seen ultrastructurally in macrophages of nine of 15 clinically normal macaques and baboons. Four of the 15 had similar granules in the intercellular spaces of the epithelium. Ultrastructurally, the eosinophilic globules were electron-dense phagolysosomes; the Feulgen-positive granules resembled nuclei of lymphocytes in various stages of pyknosis. Cytoplasmic organelles enclosed in membrane-bound vacuoles were present in the intercellular spaces of the epithelium of one monkey. Similar organelles were phagocytized by macrophages in another monkey. Feulgen-positive granules have been reported in villi of normal rodents. In all other species, including man, degenerating nuclei called "karyolytic bodies" and other evidence of enterocyte or lymphocyte degeneration have been considered abnormal concomitants of irradiation and antimitotic therapy. The significance of the findings in monkeys is not known, but they may represent a subclinical disease process. PMID- 6270870 TI - An ultrastructural study of spinal nerve roots and dorsal root ganglia in aging rats with spontaneous radiculoneuropathy. AB - The spinal nerve roots and dorsal ganglia of 104- to 135-week-old rats with spontaneous radiculoneuropathy were examined by light and electron microscopy. Demyelination was common in myelinated fibers of various diameters of both ventral and dorsal roots. The most striking alteration was wide distention of myelin sheaths, which extended throughout the entire internode. The spaces formed between separated lamellae frequently were invaded by macrophages. Subsequent vesicular degeneration of myelin seemed to be mediated by invading macrophages. These processes caused complete myelin destruction, but most axons showed no degenerative changes except for obvious reduction in diameter. Occasionally, there were clumping and partial degradation of neurofilaments and ruptured axolemma in the severely demyelinated axons. A few fibers also were undergoing wallerian-type degeneration, perhaps secondary to the severe demyelinative changes. Remyelinating fibers in various phases of repair were coexistent with markedly demyelinated ones. Demyelinative changes described above also developed within some of these remyelinated internodes. There were no remarkable changes in neurons of the dorsal root ganglia, though accumulation of lipofuscin was common. Our findings suggest that the changes in the nerve roots are essentially a primary segmental demyelination in aging rats with radiculoneuropathy. PMID- 6270872 TI - Ultrastructure of a feline extraskeletal giant cell tumor (malignant fibrous histiocytoma). AB - A subcutaneous extraskeletal giant cell tumor (malignant fibrous histiocytoma) was excised repeatedly from a 9-year-old Domestic Shorthair cat. Ultrastructurally, the mass was composed of fibroblast-like, histiocyte-like, and multinucleated giant cells, and some undifferentiated cells and mononuclear cells intermediate between the fibroblast-like and histiocyte-like cells. Fibroblast like cells were characterized by abundant well-developed rough endoplasmic reticulum, relatively smooth cytoplasmic membranes, few lysosomal structures, and finely granular chromatin. Histiocyte-like cells resembled immature macrophages. The cell membranes had many villous projections. Rough endoplasmic reticulum varied in quantity. Lysosomes were numerous. Multinucleated giant cells had characteristics of both the fibroblast-like and histiocyte-like cells. No viral particles were seen. PMID- 6270873 TI - Pox virus keratitis in a rabbit. PMID- 6270875 TI - Serology in feline leukaemia virus infections. PMID- 6270874 TI - Short communications. PMID- 6270876 TI - A malignant tumor arising from interdigitating cells; light microscopical, ultrastructural, immuno-and enzyme-histochemical characteristics. AB - A tumor in a 37 years old male is described in which the tumor cells appeared to be derived from interdigitating cells normally found in the T-cell area of lymph nodes. The patient presented with superior vena caval obstruction due to a mediastinal mass, followed by lymph node enlargement and skin lesions leading to death within 4 months. The tumor cells lacked immune markers for lymphocytic cells. They showed Ia-like antigens and high adenosine triphosphatase activity, while acid phosphatase and alpha-naphthyl acetate esterase activity was absent. Their fine morphology was strikingly similar to that of interdigitating cells. A combination of these data led us to the conclusion that this tumor represents a specific subtype of the tumors derived from the mononuclear phagocyte system, namely a sarcoma of interdigitating cells. PMID- 6270877 TI - Ultrastructure of adamantinoma of long bones. AB - Electron microscopic examination of three adamantinomas of long bones revealed structures usually observed in both mesenchymal and epithelial tumours. Tumour cells showing high alkaline phosphatase activity frequently tended to line clefts in connective tissue, resembling endothelial cells. The long processes of such cells showed fenestration. In areas showing fibre production the tumour cells were in close relationship to collagen fibrils. The latter were found, together with microfibrils, between the processes and above the basement membranes. The tumour cells were interconnected by desmosomes with tonofilaments and contained numerous bundles of microfilaments. All three cases revealed tiny intracytoplasmic inclusions resembling Weibel-Palade endothelial bodies. In addition, some of the structures in the lumena contained definite acid mucosubstances. A squamous cell pattern was present in only one of the three specimens. The coincidence of divergent structures in a single specimen has led us to the conclusion that the so-called adamantinoma of long bone might be possibly related to tumours of mesodermal or mesectodermal origin. PMID- 6270878 TI - Genetic heterogeneity among Saint Louis encephalitis virus isolates of different geographic origin. PMID- 6270879 TI - Heterogeneity of infection-associated particles in foot-and-mouth disease virus. PMID- 6270880 TI - Mapping of RNAs transcribed from adenovirus type 12 early and VA RNA regions. PMID- 6270881 TI - Mu-specific properties of lambda phages containing both ends of Mu depend on the relative orientation of Mu end DNA fragments. PMID- 6270882 TI - Characteristics of avian sarcoma virus strain PRCIV and comparison with strain PRCII-p. PMID- 6270883 TI - Two polyoma virus gene functions involved in the expression of the transformed phenotype in FR 3T3 rat cells. I. Localization of a transformation maintenance function in the proximal half of the large T coding region. PMID- 6270884 TI - Two polyoma virus gene functions involved in the expression of the transformed phenotype in FR 3T3 rat cells. II. The presence of the 56K middle-T protein in the cell membrane is not sufficient for maintenance. PMID- 6270885 TI - Activation of the alternative complement pathway by enveloped viruses containing limited amounts of sialic acid. PMID- 6270886 TI - Restriction of replication of Rous sarcoma virus mediated by the src region of the genome: analysis of the formation and integration of viral DNA and the effects of conditional and nonconditional mutations in src. PMID- 6270887 TI - the relationship of conformational changes in the Sendai virus nucleocapsid to proteolytic cleavage of the NP polypeptide. PMID- 6270888 TI - DNA topoisomerase activity associated with Rous sarcoma virus. PMID- 6270889 TI - 5-Methoxymethyldeoxyuridine-resistant mutants of herpes simplex virus type 1. PMID- 6270890 TI - Xenotropic type C virus expression in murine thymomas induced by radiation or 3 methylcholanthrene. PMID- 6270891 TI - Cleavage sites in the polypeptide precursors of poliovirus protein P2-X. PMID- 6270892 TI - Lipid-protein interactions between the surface glycoprotein of vesicular stomatitis virus and the lipid bilayer. PMID- 6270893 TI - Continuous production of Rous sarcoma virus free of transformation defective virus in clones of established RSV-transformed quail cells. PMID- 6270894 TI - Analysis of restriction endonuclease fragments of intracellular Epstein-Barr virus DNA and isoenzymes indicate a common origin of the Raji, NC-37, and F-265 human lymphoid cell lines. PMID- 6270895 TI - Expression of cell surface Friend virus gp70 does not block reinfection by ecotropic murine leukemia viruses. PMID- 6270896 TI - A virion-associated glycoprotein essential for infectivity of herpes simplex virus type 1. PMID- 6270897 TI - Identification of a primate cytomegalovirus group-common protein antigen. PMID- 6270898 TI - Isolation via transfection of feline leukemia viruses from DNA of naturally occurring feline lymphomas. PMID- 6270899 TI - A neutralizing monoclonal antibody against poliovirus and its reaction with related antigens. PMID- 6270900 TI - The effects of adenosine and guanosine on the replication of Sindbis and vesicular stomatitis viruses in Aedes albopictus cells. PMID- 6270901 TI - Partial purification of SV40 A protein and a related cellular protein from permissive cells. PMID- 6270902 TI - Alternative interactions of the SV40 A protein with DNA. PMID- 6270903 TI - Binding of dephosphorylated A protein to SV40 DNA. PMID- 6270904 TI - Structure of the genome of equine herpesvirus type 1. PMID- 6270905 TI - [Activity of various intracellular hydrolases in experimental inflammation]. AB - Activity of lysosomal hydrolases, cathepsins A and D, acid phosphatase, was induced in inflamed rat hypodermic tissue. Simultaneously, cytoplasmic enzymes leucine amino-peptidase and alkaline phosphatase were activated in the tissue. At the same time, activity of thiol-dependent enzymes/cathepsins B and C/ was decreased in all preparations studied. The data obtained suggest that both lysosomal and membrane-bound hydrolases participated in development of inflammation. Besides, proliferation of the hypodermic tissue appears to effect, by means of mediator and metabolite systems, on activity of intracellular enzymes in other tissues studied, which were not related directly to the impairment. PMID- 6270906 TI - [Changes in the structuro-functional properties of erythrocytes in vascular diseases of the brain]. AB - In patients with various impairments of brain circulation a decrease in deformation and osmotic stability of erythrocytes were observed. These findings correlated with the decrease in activity of Na+, K+- and Ca2+-ATPases from the cell membranes. At the same time, the Mg/+-ATPase activity was markedly increased. Conductivity for K+, Na+ and Ca2+ was increased in lipid bilayer of erythrocytes obtained from the patients with brain vascular pathology. PMID- 6270907 TI - [cAMP-dependent phosphorylation of myocardial troponin in experimental myocardial infarction]. AB - cAMP-dependent phosphorylation of troponin, content of cAMP and the rate of the protein kinase complex activation were studied in dog heart muscle under conditions of experimental myocardial infarction. Incorporation of 32P into troponin I as well as the content of the cyclic nucleotide were shown to decrease in the impaired muscles as compared with the normal heart muscle. In experimental myocardial infarction the rate of the protein kinase complex dissociation appears to be altered as suggested by the fact that adrenaline stimulated dissimilarly the activity of cAMP-dependent protein kinase in vitro in presence and in absence of cAMP both in the intact and necrotized muscles. PMID- 6270908 TI - [Biochemical disorders in hereditary retinal degeneration: changes in cyclic nucleotide phosphodiesterase activity and rhodopsin concentration in the retinas of Campbell rats]. AB - A correlation was noted between alterations in activity of cGMP phosphodiesterase /PDE/ and in content of rhodopsin /R/ from rats of the Campbell strain with hereditary retina degeneration as well as from control animals of the Wistar strain. Dynamics of alterations in the PDE/R ratio was similar to both in impaired and healthy animals within the first 35 days of life. These data suggest that the decrease of the PDE activity in retina of the Campbell rats occurred due to destruction and removing of outer layer of photoreceptor cell segments, i.e. it was of the secondary nature rather than of primary pathogenetic importance. After 1.5 month of the life retina aqueous extract activated PDE of cyclic nucleotides in impaired rats, but did not cause inhibition as it was observed in healthy animals. The phenomenon appears to be due to various localization of PDE inhibitors and activators in retina as well as to destruction of the layer of rod outer segments in retina of impaired rats. PMID- 6270909 TI - [Inhibition of chromatin autolysis in the process of isolating and incubating rat liver cell nuclei]. AB - Dependence of endogenous nuclear nuclease/nucleases/ activity on pH value and ion composition of the isolation and incubation media was studied. Change in buffer pH value up to pH 9.0 and addition of Ca2+ instead of Mg2+ significantly inhibited the endogenous nuclease activity in rat liver nuclei. On the basis of these data the procedure for isolation of nuclei developed by Shaveau was modified. Almost complete inhibition of the endogenous nuclease activity was achieved during destruction of chromatin DNA from the isolated nuclei in presence of exogenous nuclease/micrococcal/. PMID- 6270910 TI - [Use of fertilizers and the quality of wheat protein]. AB - The fractional and amino acid composition, as well as efficacy, net utilization and nitrogen retention ratios of protein from wheat grown under the conditions of introducing different fertilizers into the soil were studied in experiments on rats. It was found that the biological value of protein from wheat grown with the use of organic and organomineral fertilizers which were introduced into the soil on the basis of a gradual increase in their amounts appeared to exceed that from wheat harvested in the control area. Mineral fertilizers provided a higher content of protein, with its quality being virtually unchanged. The differences in the biological value confirm the respective changes in the content of protein and its fractions in the blood serum of experimental animals which received over a month the same wheat as a constituent of the balanced diets. The absence of good agreement between the chemical composition of protein and its biological value allows a conclusion about the desirability of research into the biological value of food wheat with due regard for the fertilizers introduced. PMID- 6270911 TI - [Acupuncture and various theoretical indications for its use]. PMID- 6270912 TI - [Radioisotope study of liver function in chronic pneumonia complicated by bronchial asthma]. PMID- 6270913 TI - [Morphogenesis of human rotavirus in a cell culture]. AB - Morphogenesis of human rotavirus was studied by primary green monkey kidney cell culture by electron microscopy, at the levels of the 5th and 12th passage in the cell culture. The initial stages of rotavirus morphogenesis were shown to be associated with cytoplasmic inclusions. Nucleoid or core particles formed in the periphery of the inclusions were transported by budding to endoplasmic reticulum cavities where the final stages of virion maturation occurred. The loss of the outer membrane seems to be a component part of this process. Increasing number of virus passages in the cell culture exerted a certain effect on morphogenetic processes in the infected cells. PMID- 6270914 TI - [Modeling of persistent HeLa cell infections caused by different Japanese encephalitis virus clones]. AB - The authors developed two models of persistent infections of a HeLa cell clone with mildly pathogenic clones of the Nakayama and Peking I strains of Japanese encephalitis virus. The distinguishing features of these models included the noncytocidal nature of the infectious process, predominance of the small-plaque phenotype, further decrease of the infectious properties of the persisting viruses. Altogether during the observation period 84 subpassages of each of the two systems were made without any visible signs of cell degeneration. PMID- 6270915 TI - [Role of genetic disorders in the etiology of nephroblastoma]. PMID- 6270916 TI - The pattern of ectopic hormone production in lung cancer. AB - Pulmonary cancers produce many hormonal polypeptides. There is a tumor-specific pattern to the appearance of abnormal adrenal function and inappropriate secretion of vasopressin, which are frequently found in small cell undifferentiated carcinoma but occur only very rarely, if at all, in squamous tumors. Humoral hypercalcemia, on the other hand, occurs almost entirely in squamous tumors and is rarely if ever seen in small cell or large cell tumors or in adenocarcinoma. In contrast, "big ACTH" and beta lipotropin are found in the plasma and tumor extracts of lung cancers of all types. Calcitonin and the beta chain of human chorionic gonadotropin are also found in the plasma of a considerable portion of patients with all histological types of lung cancers. PMID- 6270918 TI - The role of radiation therapy for carcinoma of the lung. AB - A large proportion of patients with carcinoma of the lung may benefit from the use of radiation therapy. Operable patients have not been shown to benefit from preoperative irradiation, but postoperative irradiation has improved survival in those found to have involvement of hilar or mediastinal lymph nodes. Radiation therapy is the only potentially curative treatment for patients who are inoperable, but do not have distant metastasis. Control of the local tumor is very dependent upon dose-fractionation-time relationships. Patients who are relatively asymptomatic, i.e., they have a high performance status, are curable if treated promptly with radiation therapy. Small cell carcinoma requires both radiation therapy and chemotherapy. The optimal method of combining the two modalities is yet to be determined, but prophylactic cranial irradiation is necessary to control microscopic metastases that are not affected by systemic chemotherapy, and thoracic irradiation is necessary to give the highest probability of control of the primary tumor. Prophylactic cranial irradiation has also been shown to reduce the frequency of brain metastasis in patients with squamous carcinoma, large cell carcinoma, and adenocarcinoma; it may become more important in these cell types when more effective chemotherapy is developed. PMID- 6270917 TI - In vitro study of the biology of small cell carcinoma of the lung. AB - WE HAVE DEVELOPED THREE TYPES OF EXPERIMENTAL SYSTEMS FOR THE STUDY OF SCCL: (1) serially heterotransplanted tumors in athymic nude mice; (2) continuous, clonable cell cultures; and (3) direct clonogenic assays for tumor specimens. These systems have their own individual advantages, applications, and limitations, but these are interrelated and complementary. The study of these systems has greatly aided our understanding of the biology of SCCL, and its relationship to other lung cancers and the APUD cell system. In addition, new markers for SCCL have been identified, such as a creatine kinase and its BB isoenzyme (CK-BB). These cellular markers may have clinical applications, as serum levels of CK-BB are an indicator of tumor burden. Assays for clonogenic tumor cells may permit selection of optimal drug combinations for the treatment of individual tumors. Variant cultures having the morphology of SCCL, but lacking some or all of the other features, have been identified. While our systems have been used primarily for biological studies, they have clinical applications for both diagnostic and therapeutic purposes. PMID- 6270919 TI - Causes of treatment failure and death in carcinoma of the lung. AB - Studies of patterns of failure and causes of death have been undertaken based upon the WHO histopathologic classification. In a randomized trial of thoracic irradiation +/- chemotherapy (hydroxyurea and CCNU), patterns of failure did not seem to differ by cell type; the largest group was "death without progression." A subsequent clinical trial of thoracic irradiation +/- cranial irradiation permitted a more detailed evaluation. Patients with squamous cell carcinoma had a higher rate of local failure than distant metastasis. Those with small cell carcinoma had a lower local failure rate and a high rate of distant spread. Patients with adenocarcinoma and large cell carcinoma had the lowest local failure rate, but had a high rate of distant metastasis. In 300 consecutive patients with autopsies, 75 percent with squamous carcinoma died of complications of the thoracic tumor and only one-quarter had extrathoracic dissemination; 30 percent with small cell carcinoma died of local tumor complications and 70 percent had carcinomatosis; 40 percent of patients with adenocarcinoma and large cell carcinoma died of intrathoracic complications, and 55 percent had distant metastases. Half the patients with small cell carcinoma, large cell carcinoma, and adenocarcinoma had brain metastases at autopsy. Future clinical trials should emphasize better control of the most common sites of failure. PMID- 6270921 TI - Efficiency of conventional versus marker technique for shortening the time in digestion trials. AB - Investigation was carried out to determine the digestibility of the main nutrients (DM, CP and energy) in digestion trials using some roughages in order to reduce the collection period and to study the suitable time of the marker technique and its application. This study revealed the following results: (1) The average recovery of chromic oxide in feces were 99.71 +/- 3.75% in the 3rd day (after administrating the marker), 95.17 +/- 1.03% in the 5th day and 96.53 +/- 2.35% in the 7th day. It could be concluded that this marker is indigestible and could be used as an ideal one. (2) There were no significant differences between the digestibility values with the conventional method and those of the two short period techniques (the first three days and the first five days collection periods). The percentage differences were not exceeding 6% except some deviations with CP digestibility in one trial (hay : rice straw mixture). These two methods seemed to be reliable in practice. (3) The same differences were found between the digestion coefficients, which conducted from the conventional method, and those of the marker technique using three times for collection except high deviations were found with one trial in the last two procedures. Using marker technique appeared promising from the 3rd day up to the 7th day after its administration. PMID- 6270920 TI - [Aggregation and separability of the shikimate pathway enzymes in yeasts]. AB - Gel filtration was employed to estimate the molecular weights and to determine possible physical aggregation of enzymes [5-dehydroquinate synthase (DHQ synthase), 5-dehydroquinase (DHQase, EC 4.2.1.10), shikimate: NADP oxidoreductase (EC 1.1.1.25), shikimate kinase (EC 2.7.1.71), 3-enolpyruvylshikimate 5-phosphate synthase (EPSP synthase)] in the shikimate pathway in eleven species of yeasts. The five enzymes were not aggregated in extracts of Hansenula henricii, H. fabianii, H. anomala, Candida utilis, Pichia guilliermondii, and Lodderomyces elongisporus. Two enzymes (DHQase and shikimate:NADP oxidoreductase) were not separable by this method and by ion exchange chromatography, and we conclude that they exist as an aggregate in these yeasts. Evidence is presented for an enzyme aggregate containing five activities, with a molecular weight of approximately 280,000 in Rhodosporidium spaerocarpum, Rh. toruloides, Rhodotorula rubra, Saccharomycopsis lipolytica, and Saccharomyces cerevisiae. Similarities between the enzymes in the shikimate pathway of plants, bacteria, and other fungi and those of investigated yeasts are discussed. PMID- 6270922 TI - [Alcohol and central and peripheral nervous system disorders]. AB - It belongs to the definition of chronic alcoholism that also clinically established lesions of the organs can be proved. As to the nervous system the case in question are multilocular defects which concern the central and peripheral nervous system. The local accentuation, the developmental speed and the intensiveness of the alcoholic defect of the cellular metabolism lead to different courses. The suggestion of their individuality must at the same time take into consideration the character of chronic alcoholism which on principle affects the whole nervous system. PMID- 6270923 TI - [The diagnostic value of M-mode-, two-dimensional echocardiography, and computed tomography in comparison to the results of cardiac catheterization in the diagnosis of tumors of the heart (author's transl)]. AB - 12 patients with cardiac tumors (7 myxomas of the left, 2 of the right atrium, 2 ventricular, 2 intramural tumors and 1 epicardial cyst) were examined by M-Mode (ME) and two-dimensional (2DE) echocardiography, computertomography (CT) and heart catheterization (HK) with angiography (A). The results were compared with intraoperative findings. There were no false negative results with 2DE and CT, but in 1 case the size of an intramural tumor was underestimated by 2DE. With ME all atrial myxomas were diagnosed, but 2 ventricular and 2 intramural tumors as well as the epicardial cyst were overlooked. Also with HK and A, the 2 intramural tumors and the epicardial cyst were not recognized well. The results indicate that the diagnosis of cardiac tumors can be made with great accuracy by noninvasive methods. The sensitivity of the single methods depends on the location of the tumors. PMID- 6270925 TI - [Diagnostic significance of calcifications in the mammogram]. AB - Since microcalcifications occur in mammary carcinoma in a quite specific manner and above all very early, i.e. long before an infiltrative focal shadow can be recognized in the mammogram, the microcalcifications have proved to be a significant and in many cases the only early symptom of small, clinically still occult mammary carcinomas. On the basis of microcalcifications, we are so far been able to detect a total of 220 clinically unequivocal occult small and very small mammary carcinomas with an average tumor size of 6 X 8 mm, and were thus able to apply the necessary and decisive early therapy. Today, microcalcifications enable us to detect to an increasing extent even preinvasive stages of mammary carcinoma and also to verify them by biopsy. PMID- 6270924 TI - [Determination of platelet aggregation using the KZM-1-R meter (aggregation inducers: ADP, collagen, ristocetin, ristomycin, adrenaline. Therapy control by use of acetylsalicylic acid)]. PMID- 6270926 TI - [Cyclic AMP- and Ca-dependent phosphorylation of functionally significant proteins in presynaptic structures]. PMID- 6270928 TI - [Cocarcinogenic action of vaccinia and herpes simplex viruses in tissue culture]. PMID- 6270929 TI - [Intensive combined chemotherapy and radiation therapy of small cell lung cancer]. PMID- 6270927 TI - [Role of adrenoreceptors in regulating the efficiency of adrenergic synaptic transmission]. PMID- 6270930 TI - [Colorectal carcinoma in patients under 35 years of age (author's transl)]. PMID- 6270931 TI - [Liver tumours in pregnancy, following previous intake of hormonal contraceptives (author's transl)]. AB - Liver tumours developed in two women during pregnancy which followed discontinuation of hormonal contraception. Liver cell adenoma developed in a secundipara, 24 years of age, and its rupture during puerperium led to death by bleeding, after surgical intervention had failed. Hepatocellular carcinoma was recorded from a secundipara, 22 years of age, and surgically treated, following premature delivery. Problems of diagnosis and the need for interdisciplinary cooperation are discussed and compared with literature data. The pathogenesis of liver tumours in the wake of exogenous hormone inflow and in the context of subsequent pregnancy is another topic of discussion. PMID- 6270932 TI - [Comparative serological studies of blood and milk in the diagnosis of enzootic bovine leukosis]. PMID- 6270933 TI - [Glycogen content and glucose-6-phosphatase activity in the ligated frog ischiatic nerve]. AB - Glycogen content and glucose-6-phosphatase activity were determined in the both proximal and distal parts of the ligatured ischiadicus nerve of the frog. The results show that endogenous glycogen stores (or derived from them glucose) can move along the nerve and can be utilised in ischiadicus nerve metabolism. These data support our earlier findings that glycogen biosynthesis depends on glucose-6 phosphatase activity. PMID- 6270934 TI - [Phosphatase-negative Staphylococcus epidermidis]. AB - As a result of the study of a number of additional characteristics, more than a half of the cultures, formerly identified as phosphatase-negative S. epidermidis according to the scheme of the International Subcommittee on the Taxonomy of Staphylococci and Micrococci, have been shown to be more similar to S. Hominis and S. captis on account of their properties. The study of correlation between the main differentiating characteristics of S. epidermidis has shown that it is poorly pronounced. For this reason the decision to classify a phosphatase negative culture with S. epidermidis should be based on the study of additional characteristics which differentiate this species not only from S. saprophyticus, but also from other phosphatase-negative staphylococci. PMID- 6270935 TI - [Mechanism of the antimicrobial action of myeloperoxidase. The role of adsorption of the enzyme on the target cell surface]. AB - Myeloperoxidase (MPO), obtained from the granular fraction of bovine neutrophils at low concentrations (3 nm), exerted antibiotic influence on E. coli. After treatment with the MPO3 - H2O2 - Cl- system the number of living cells in the suspension dropped to 4.7% of the initial value. The preliminary treatment of MPO with antibodies at high concentrations, thus preventing MPO from being adsorbed on the surface of bacterial cells without suppressing its enzymatic catalytic activity, resulted in the considerable decrease of its bactericidal effect. These results suggest that the adsorption of MPO on the surface of target cells is the essential condition of its antimicrobial action. PMID- 6270936 TI - [Effect of reflexotherapy on central and peripheral hemodynamics in patients with the radicular syndrome of lumbar osteochondritis]. AB - The central and peripheral hemodynamics was examined in patients with the radicular syndrome of lumbar osteochondrosis who were treated by reflexotherapy methods. In addition to the direct favourable effect of microacupuncture and classical acupuncture on the circulatory system a longer duration of the positive effect of microacupuncture was revealed. An interconnection between the favourable changes of the patient's state and the improvement of the systemic and local circulation was noted. PMID- 6270937 TI - [Status of active and passive cation transport through cerebral synaptic membranes during seizures]. PMID- 6270938 TI - [Opiate receptors of peripheral blood lymphocytes (effect of morphine and naloxone on human T-lymphocytes)]. AB - Using the method of spontaneous rosette formation the authors studied the effects of morphine and naloxone (a specific morphine antagonist) on T-lymphocytes of man peripheral blood. Treatment of the lymphocytes with morphine led to a diminution of the number of rosette-forming cells. Naloxone did not influence the rosette formation process, but eliminated the specific effect of morphine. The opiate receptors on the thymus-dependent lymphocytes may be an instrument of hormonal regulation of cell interaction processes in individual lymphocyte populations. PMID- 6270939 TI - [Recently registered drugs]. PMID- 6270940 TI - Two cases of 17 alpha-hydroxylase deficiency--one combined with complete gonadal agenesis. AB - Two cases of 17 alpha-hydroxylase deficiency are described. Both patients had primary amenorrhoea, total lack of female secondary sexual characteristics, slight hypertension and hypokalaemia. One patient was of male genotype (male pseudohermaphrodite), and in addition this patient had complete gonadal agenesis. The other patient was of female genotype. In both patients the level of plasma corticosterone was markedly increased, whereas the concentration of plasma cortisol was very low and plasma aldosterone low within the normal range. Furthermore, the plasma ACTH level was significantly increased and the plasma renin activity around the lower normal limit. The urinary excretion of corticosterone metabolites was markedly increased, whereas the excretion of both cortisol metabolites and tetrahydroaldosterone was decreased. The patients had no symptoms of glucocorticoid deficiency. Treatment with dexamethasone 0.5 mg daily completely suppressed the abnormal corticosterone production and normalized both blood pressure and serum potassium. In addition, the patient of male genotype has received sequential therapy with oestrogen and gestagen for 3 years, but so far no development of the secondary sexual characteristics has occurred. PMID- 6270941 TI - Virilization of a post-menopausal woman by a testosterone-secreting Leydig cell type adrenal adenoma. PMID- 6270942 TI - Characterisation of the cyclic AMP response to thyrotrophin in monolayer cultures of normal human thyroid cells. AB - The cyclic AMP response to thyrotrophin (TSH) has been investigated in cells prepared from human thyroid tissue obtained during surgery for subtotal laryngectomy, and maintained under in vitro conditions as primary monolayer cultures. When cells were incubated with 1.0 mU TSH/ml, a maximal level of intracellular cyclic AMP was reached after 20 min of incubation in the presence of 0.5 mM 3-isobutyl-1-methyl xanthine (MIX). This level of cyclic AMP was sustained for at least 2 h. Half-maximal stimulation of cyclic AMP was produced by TSH doses of between 1 and 5 mU/ml. In a study of a series of eight groups of monolayer cultures, each derived from a single, different thyroid gland, the mean stimulation of cyclic AMP given by 50 mU TSH/ml was 37.8-fold greater than in non stimulated cell monolayers. Significant stimulation of 50 microunits TSH/ml was observed in some monolayers and the precision of measurement of TSH was better than 15% over the TSH dose range 0.2-1.0 mU/ml. The magnitude of the cyclic AMP response to TSH was unaffected by the presence in the incubation medium of 20% (v/v) normal human serum. A cyclic AMP response to TSH was still demonstrable in cells that had been maintained for a period of 22 days in monolayer culture, although the response was reduced in comparison with that given by 4-5 day old cultures. PMID- 6270943 TI - Development and regulation of the prostaglandin F2 alpha receptor in the rat ovary. AB - Ovaries of adult rats specifically bind PGF2 alpha while those of immature animals do not. Induction of luteinization by hCG in juvenile animals, however, results in specific binding of PGF2 alpha. It is suggested that luteal cells are the only cell type of the ovary, which is endowed with specific receptors for PGF2 alpha. The number of PGF2 alpha binding sites varies during the ovarian cycle. Most free receptors are detectable in early pro-oestrus, least in the oestrus stage. The oscillation of receptors disappears after inhibition of prostaglandin synthesis by indomethacin. Therefore the apparent cyclic variation of prostaglandin receptors must be ascribed to occupancy of the receptors by varying amounts of endogenous prostaglandin F2 alpha. PMID- 6270945 TI - A case of hemolytic anemia due to erythrocyte pyrimidine 5'-nucleotidase deficiency. AB - A propositus, the offspring of a first-cousin marriage, was presented with severe hemolytic anemia, splenomegaly, jaundice, and growth retardation. Marked basophilic stippling of erythrocytes was shown by Wright's stain. Erythrocyte 5' nucleotidase activity was found markedly decreased, whereas red blood cell glucose-6-phosphate dehydrogenase activity was elevated as the reduced glutathione level. His growth and anemia improved following splenectomy. His sister was also similarly affected. PMID- 6270944 TI - Distribution of prostaglandin E2 and prostaglandin F2 alpha receptors in human myometrium. AB - Human myometrium was studied for specific binding of PGE2 and PGF2 alpha. PGF2 alpha-binding was almost undetectable, but specific binding sites for PGE2 with high affinity (KD = 2.7 +/- 0.4 x 10(-9) M were demonstrated. The binding capacity for PGE2 exhibited a topically different distribution pattern with the highest values in the central parts and low to undetectable levels in the cervical region. Binding characteristics were analyzed by receptor kinetics, revealing a homogeneous receptor population. Binding capacity in uteri obtained from post-menopausal women was of the order of 900-940 fmol/mg protein. Oestrogen pre-treatment and pregnancy were associated with a 3-fold reduction of the PGE2 binding capacity. PMID- 6270946 TI - [Ultrastructural studies and neutrophil myeloperoxidase determination in rats with acute 2,4-dichlorphenoxyacetic acid poisoning]. AB - The aim of the work was to study changes in the ultrastructure and activity of different developing forms of cells in the neutrophil series in rat bone marrow during acute poisoning with 2,4-dichlorphenoxyacetic acid. The results of the study enabled determination of the influence of this herbicide on the process of granulogenesis in the neutrophils to be achieved. Disturbances of granulogenesis appeared in some cells of the experimental group II (160 mg of 2,4-D-acid/kg of body weight) and in most cells in the experimental group III (400 mg of 2,4-D acid/kg). The disturbances manifested themselves mainly as a decrease in the number of primary and secondary granules in different developing forms of the neutrophil series. This decrease was associated with appearance of numerous cytoplasmic vacuoles. The results show that primary granules participate in phagocytosis. PMID- 6270947 TI - [Effect of mitogens on the changes in intracellular cyclic AMP concentration in the lymphocytes of healthy persons and patients with lymphoblastic leukemia]. AB - In the lymphocytes of 20 healthy subjects and 17 patients with chronic lymphatic leukaemia (CLL) the cAMP level was determined before and at different time periods during cell incubation with mitogens (phytohaemagglutinin--PHA, concanavalin A--Con A, and pokeweed mitogen PWM). The experiments demonstrated that PHA-induced lymphocyte proliferation was associated in healthy subjects during the first 10--20 minutes of incubation with a rise in the intracellular cAMP concentration, while the level of cAMP decreased systematically during 30 minutes of incubation with Con A and PWM. Incubation of lymphocytes of the patients with CLL during 30 minutes with PHA, Con A and PWM caused no changes in the low cAMP level in these cells. The obtained results suggest that PHA activation of lymphocytes in healthy subjects is connected with changes in the intracellular concentration of cAMP. PMID- 6270949 TI - [Pathomorphological study on the process of the generation of an experimental ovarian tumor by intra-spleen ovarian autografting in a rat (author's transl)]. PMID- 6270948 TI - [Effects of exogenous gonadotropin on the generation and growth of an ovarian tumor by intra-spleen autografting in a rat (author's transl)]. PMID- 6270950 TI - Cortisol, glucose, and hemodynamic responses to surgery after naloxone administration. AB - The adrenocortical, hyperglycemic, and hemodynamic responses to cholecystectomy or vagotomy were studied in 16 patients under halothane and N2O/O2 anesthesia. The patients were randomly divided into two groups: eight patients received naloxone in doses used clinically (2.5 microgram/kg i.v.) just before induction of anesthesia, while eight subjects received placebo. The results showed insignificant differences in plasma concentrations of cortisol and glucose between groups. Nor did blood pressure, heart rates, and inspired halothane concentrations differ significantly between groups. Thus, inhibition of opiate receptors and endorphins by naloxone in an otherwise clinically effective dosage does not influence the adrenocortical, hyperglycemic, or hemodynamic responses to surgical stress. We therefore conclude from our data that opiate receptors and endorphins are not involved in the initial phase of the endocrine-metabolic responses to surgery nor are they part of the neural mechanisms mediating stress induced analgesia. PMID- 6270951 TI - A comparative histochemical mapping of acid phosphatase, 5-nucleotidase and non specific esterase in the olfactory bulbs of rabbit and hedgehog. AB - The paper deals with comparative account of the distribution of acid phosphatase 5-nucleotidase and non-specific esterase in the olfactory bulbs of rabbit and hedgehog. The acid phosphatase is observed in all the neurons of both the animals. However, the concentration of the enzyme is higher in hedgehog as compared to rabbit. The intensity of 5-nucleotidase in all the layers of olfactory bulb of hedgehog is stronger than in the rabbit. Intensity of non specific esterase is quite higher in the olfactory bulb of rabbit than of hedgehog. Along with comparison the distribution of these enzymes have been correlated with their role in the olfactory senses. PMID- 6270952 TI - [Cerebral angiophotoscintigraphy in the screening of cerebral vascular diseases]. PMID- 6270953 TI - [Familial glomus tumors: clinical and genetic aspects]. PMID- 6270954 TI - A "reverse" solid-phase radio-immuno-assay for IgM-antibodies to hepatitis A virus. AB - A "reverse"solid-phase radio-immuno-assay for IgM antibodies to hepatitis A virus (HAV) was developed. Anti-human IgM immunoglobulins were bound on the wells of polyvinylchloride microtiter plates. Serum specimens were incubated in the anti human IgM coated wells and bound IgM antibodies were then assayed for antigen specificity by subsequent incubations with HAV antigen and 125I-labelled human anti-HAV IgG. The test showed a high sensitivity and specificity for anti-HAV IgM antibodies. No false-positive reactions were observed either in the sera from patients with hepatobiliary disorders other than HAV infection or in the sera containing both rheumatoid factor and anti-HAV IgG antibodies. In acute HAV infections specific IgM antibodies were present already in the first specimens taken within a few days after the onset of jaundice. The persistence of the IgM antibodies was from 4 to 6 months. IgM antibody titers up to 1,000,000 were observed in the acute phase of HAV infection. In routine diagnostic work the titration of the sea was not necessary, since a reliable qualitative result was obtained by testing the sera in a single dilution of 1:100. A similar "reverse" immuno-assay principle may be adaptable for the diagnostic determination of IgM antibodies to different viral and microbial antigens. PMID- 6270956 TI - Effect of age and route of administration on LD50 of lithium chloride in the rat. AB - Male inbred rats were used to determine the lethal dose of lithium chloride. The rats were 3 weeks, 6 weeks, 3 months and 6 months old, and lithium chloride was given intraperitoneally, subcutaneously and orally. In relation to age, LD50 was significantly higher in rats of 6 weeks after oral administration than LD50 in rats of 3 and 6 months. No differences were found following intraperitoneal and subcutaneous administration. In relation to route of administration, LD50 was significantly higher in rats of 3 and 6 weeks after oral administration than LD50 after intraperitoneal and subcutaneous administration. This difference was not found in rats of 3 and 6 months. It is concluded that age and route of administration are of significance for LD50 of lithium chloride in the rat. PMID- 6270955 TI - Effect of cGMP derivatives on contraction relaxation cycle, release of norepinephrine and protein kinase activity in guinea pig vas deferens. AB - Contractions of the guinea pigs vas deferens were induced by field stimulation and the effects of derivatives of cGMP were tested on the contraction-relaxation cycle. Relaxation was induced by 8-Br-cGMP and O2'-mb-cGMP. On the contrary, db cGMP and N2-mb-cGMP potentiated the contraction. The relaxant effect of 8-Br-cGMP was not mimicked by 8-Br-5'GMP, instead the latter caused the opposite effect. Since neither 8-Br-cGMP nor db-cGMP affected the release of norepinephrine on field stimulation the cGMP derivatives induced their effects at a postsynaptic site. 8-Br-cGMP was found to be 100-fold more potent than cGMP to simulate protein kinase activity in crude extracts from guinea pig was deferens. N2-mb cGMP and O2'-mb-cGMP were approximately equipotent to cGMP while db-cGMP was less active. 8-Br-5'-GMP did not stimulate the protein kinase. It is suggested that the results support the hypothesis that cGMP acts as a relaxant in the smooth muscle of the vas deferens. The potentiation caused by db-cGMP and N2-mb-cGMP on contractions could be dependent on the substitution in the guanine moiety of the molecules with the butyryl group. Their effect was thus probably unrelated to a specific cGMP effect. PMID- 6270957 TI - Hormonal and neurogenic adrenergic control of the fluid transfer from skeletal muscle to blood during hemorrhage. AB - During hemorrhage net transcapillary absorption of interstitial fluid from skeletal muscle into the intravascular space compensates effectively for the blood loss. This absorption of fluid is mainly linked to decrease of the capillary hydrostatic pressure (Pc), as caused by reflex adrenergic re-adjustment of the ratio of pre- to postcapillary resistance (ra/rv). The present study demonstrates the existence of both a neurogenic and a humoral component in the adrenergic control of the fluid transfer for skeletal muscle to blood. In the early period of bleeding (less than 5 min) reflex activation of the vasomotor fibres contributed significantly to the fluid absorption. The subsequent, main part of the fluid gain from the extra- to the intravascular space was due to the action of the blood-borne catecholamines. Both the neurogenic and the hormonal control of the fluid absorption process was mainly linked ot beta-adrenergic inhibition of vascular smooth muscle tone. This control was effected via two mechanisms, viz. by a relatively larger beta-adrenergic dilation of post- than precapillary resistance vessels, leading to adjustment of ra/rv and thereby to decrease of Pc, and via beta-adrenergic dilation of "precapillary sphincters' leading to increased capillary surface area available for fluid exchange. PMID- 6270958 TI - Further studies on beta-adrenergic control of transcapillary fluid absorption from skeletal muscle to blood during hemorrhage. AB - During hemorrhage fluid is absorbed from the extravascular space of skeletal muscle into the intravascular compartment in order to compensate for the blood loss. In a previous report (Lundvall & Hillman 1978) this process was found causally linked to reflex, beta-adrenergic control of the capillary hydrostatic pressure (via adjustment of pre-/postcapillary resistance) and of the capillary exchange area (via adjustment of "precapillary sphincter' tone). The present study extends the experimental evidence for such beta-adrenergic control of plasma volume in bleeding, showing that it operates for prolonged periods of time and in a graded manner over a wide range of hemorrhagic hypovolemia. The investigation also provides information about the quantitative importance of the fluid gain from muscle to blood for plasma volume restoration in hemorrhage. Thus, during moderate bleeding (10 ml x kg b.wt.-1) the cumulative (45 min) fluid gain from muscle to blood averaged 1.4 ml x 100 g muscle. -1 This implies, when calculated for the total muscle mass, that about 2/3 of the shed blood volume will be replaced by interstitial fluid within less than one hour. PMID- 6270959 TI - Development and validation of a radioimmunoassay for beta-endorphin-related peptides. AB - A radioimmunoassay method suitable for measuring levels of B-endorphin, B lipotropin and proopiocortin in tissue and plasma extracts was developed and the method was evaluated by using 3 independently prepared antisera. Of the several variables tested the choice of assay buffer and test tubes and the purification of tracer were found to be the most critical in the successful performance of B endorphin radioimmunoassay. The prevention of degradation of tracer during incubation was also necessary when crude tissue extracts or plasma were assay. The sensitivities of the assays obtained with the 3 antisera utilized (Bendo 2, K2 and RB 100) were 1, 2.8 and 4 fmol B-endorphin per tube. All the antisera crossreacted equimolarly with B-lipotropin. The method was employed to measure the levels of B-endorphin immunoreactivity in rat pituitary and plasma by separating the different immunoreactants by gel filtration. It was found that both pituitary and plasma contain significant amounts of proopiocortin, B lipotropin and B-endorphin, the molar proportions being 10:33:57 in pituitary and 15:15:17 in plasma, respectively. Both anterior and posterior lobes of rat pituitary were found to contain all the three immunoreactants. However, anterior lobe contained mostly the larger molecules, while posterior lobe was rich in B endorphin. No absolute levels of the immunoreactants could be obtained due to the heterologous system used. Moreover the elution pattern of the immunoreactivity was found to be dependent on the conditions used for elution in gel filtration: higher proportion of the immunoreactivity eluted like B-endorphin when the elution was done in dissociating conditions (6 M urea) compared to elution with ordinary buffers. PMID- 6270960 TI - [Beta-lactamase inhibitors. I. Synthesis and biological properties of penicillanic acid derivatives]. PMID- 6270961 TI - Sensitising effect of misonidazole on NHIK 1922 cells irradiated in vitro or as solid tumour in athymic nude mice. AB - The sensitising effect of the hypoxic cell sensitiser misonidazole (MIS) on NHIK 1922 cells irradiated either as single cells in vitro or as tumours in vivo in air-breathing and in dead mice was investigated with subsequent determination of in vitro colony-forming ability. In absence of MIS no significant difference was found between the dose-response curves after irradiation of tumours in dead mice and of cells in suspension under hypoxic conditions. After irradiation of tumours in air-breathing mice the same enhancement ratio of 1.4 was found whether MIS was administered in doses of 250 or 1000 micrograms/g body weight. In dead mice the enhancement ratio was 1.3 and 2.6 after 250 and 1000 micrograms/g, respectively. The sensitising effect of MIS for cells irradiated in suspension under extremely hypoxic conditions corresponded to that found in dead mice when a comparable concentration of MIS was used. PMID- 6270962 TI - Postmenopausal bone mineral loss after treatment for malignant gynaecologic tumours. AB - Bone mineral content after treatment for malignant gynaecologic tumours was examined in women of fertile age. The treatment given was intracavitary radium applications followed by bilateral oophorectomy. In women who received oestrogen substitution the bone mineral content increased by one per cent a year, whereas nonsubstituted women lost 3 per cent of their initial bone mineral content per year. The findings suggest that oestrogen deficiency is the predominant cause of bone mineral loss initiated by this therapy. PMID- 6270963 TI - Long-term effect of postoperative irradiation on the immune functions in patients with mammary carcinoma. AB - The effect of postoperative irradiation on the immune functions of 13 patients with breast carcinoma is reported, using as parameters the peripheral blood lymphocyte count, percentages of E and EAC rosette forming cells, and lymphocyte proliferative responses to PHA, Con A and PPD. After irradiation the number of peripheral blood lymphocytes was reduced during 8 months. The percentages of E and EAC rosette forming cells were not altered during the observation time of 7 to 36 months. In the proliferative responses of lymphocytes to PHA, Con A and PPD, the postoperative irradiation caused a decrease which, regarding PHA and Con A, lasted up to 8 months and then recovered. The decrease in the proliferative responses to PPD was stronger and lasted during the whole observation time. In the mitogenic responses of patients with recurrent or disseminating disease no difference could be demonstrated as compared with the patients living recurrence free. PMID- 6270964 TI - Effect of exogenous angiotensin-II on local blood flow in kidneys with neoplasm. Experiments in the rat. AB - The effect of angiotensin-II on the local renal blood flow and arterial blood pressure was investigated in the rat with the H2 gas washout technique. Increasing intravenous infusion rates gave a decreasing blood flow and increasing blood pressure, renal vascular resistance being close to proportional to the log of infusion rate. In kidneys with experimental neoplasm, the flow reduction was proportionally less in the tumor than in intact renal tissue. The respective flow levels were maintained for 10 to 45 min without tendency to change. PMID- 6270965 TI - Influence of 90Sr-contaminated milk on the ovaries of foetal and young mice. AB - A dose of 185 kBq (5 microCi) 90Sr was intravenously injected into pregnant CBA mice on the 19th day post coitum. Four groups of female young were examined when aged 56 days: (1) non-contaminated females (controls), (2) only exposed in utero, (3) only having suckled contaminated milk and (4) exposed to 90Sr in utero and having suckled contaminated milk. The effect of the nuclide was expressed as a comparison of the number of remaining cells in the ovaries, totally and in different stages of development, between 90Sr-contaminated young and controls. Young mice that suckled contaminated milk had a reduced number of cells compared with the controls. The ovaries of in utero contaminated females were injured to a less severe degree if the individuals were given noncontaminated milk, compared with those suckling contaminated dams. Minute changes of the body burden of 90Sr were especially reflected in the number of young oocytes, being the most radiation sensitive stage during the ovarian development. PMID- 6270966 TI - Adjuvant radiation therapy compared with cyclic chemotherapy in patients with mammary carcinoma. I. Changes of blood lymphocyte subpopulations. AB - Peripheral lymphocyte subpopulations were examined in 37 women with breast carcinoma who were postoperatively randomized to adjuvant local radiation therapy or cyclic treatment with 5-Fluorouracil, Methotrexate and Chlorambucil. Both T and non-T lymphocyte counts were reduced but the latter subpopulation was reduced to the highest relative extent in both groups of patients. Following radiation therapy the non-T cells repopulated more rapidly than the T cells, whereas the repopulation seemed to be the reverse in patients treated with chemotherapy. PMID- 6270967 TI - Electrets for use as dosimeters in radiography. AB - A compact device for the manufacturing of electrets and for the measurement of the dose to the skin of the patient in radiography has been developed and described. The surface potential of a polarized polymer electret decreases when it is exposed to radiation. The measurement of the variation in surface potential appears to be a reliable method for the evaluation of the dose. The electret does not disturb the examination nor the radiographic image as it can be given tissue equivalent properties. PMID- 6270968 TI - Electron beam dose planning using discrete Gaussian beams. Mathematical background. AB - A general method for computerized electron beam dose planning is proposed based on the use of a square or hexagonal grid or matrix of discrete Gaussian elementary beams. Using this method the distribution of absorbed dose in an electron beam of arbitrary cross-section incident on an irregular patient surface can be calculated taking the influence of tissue inhomogeneities into account. The derived expressions for the absorbed dose in narrow and broad uniform beams are compared with experimental data and Monte Carlo calculations. Good agreement is obtained over the clinically significant portion of the absorbed dose distribution at a fairly wide grid spacing of about 0.5 cm. PMID- 6270969 TI - Invasive squamous cell carcinoma of the uterine cervix. II. Reproducibility of a histopathologic malignancy grading system. AB - A histopathologic malignancy grading system for invasive squamous carcinoma of the uterine cervix has previously been presented. The grading results in a total malignancy point value for every patient (range 8-24). This score has been proven to have a significant prognostic value for the individual patient. The inter observer reproducibility of this system has been examined by two pathologists, on the pretreatment biopsies of 100 patients and the intra-observer reproducibility on 190 patients (observer one) and on 100 patients (observer two). These biopsies were estimated twice and three times, respectively, without knowledge of former results and at an interval of 2 to 6 months. The inter-observer correlation coefficient was 0.54, the intra-observer correlation coefficients 0.94 and 0.74, respectively. PMID- 6270970 TI - Megavolt electron irradiation for localized mycosis fungoides. AB - A retrospective analysis was carried out to evaluate the effectiveness of small field megavolt electron irradiation for localized mycosis fungoides. Only local field electron beam therapy was employed for limited disease reserving total skin electron irradiation for multiple lesions or diffuse disease covering at least 25 per cent of the entire body surface. Of the 14 patients with limited disease treated between 1964 and 1973 with the local field technique, 10 patients (71 per cent) are alive without evidence of disease at a minimum of 5 years. In contrast, of 200 patients with extensive cutaneous disease who received total skin electron irradiation, only 16 (8 per cent) were considered cured. It is concluded that early localized mycosis fungoides is potentially curable, and that limited field electron beam therapy with a relatively low total dose is adequate to obtain excellent response. PMID- 6270971 TI - Irradiation of malignant tumors of the parotid gland. AB - The treatment results in a series of 30 patients with parotid gland carcinoma are reported. Postoperative irradiation to the parotid bed and regional lymph node appears to improve the result of treatment. No difference in the results or complications was found between the group of patients treated by external irradiation or external irradiation combined with radium implantation. Irradiation of residual disease was more effective than irradiation of the recurrences. PMID- 6270972 TI - Hodgkin's disease irradiated with the inverted-Y technique. AB - Thirty-four patients with Hodgkin's disease were treated with the inverted-Y technique. Target absorbed dose was 40 Gy given in a split-course schedule to all but one patient. Only one recurrence occurred. Seventeen patients remained symptom-free after treatment and another 9 were rescued by further therapy. Eight patients died. All 7 patients in stages I and II are alive. In stage III A more patients with upper abdominal disease remained symptom-free after treatment than patients with lower abdominal disease. Of 7 patients in stage III B, 5 died. Actuarial survival at 10 years was significantly better for patients without systemic symptoms. Radiation side effects were mild. One serious complication occurred, acute gastrointestinal ulceration in the patient given the total dose in one series. No permanent symptom-producing side effects from liver, kidneys, spinal cord or bone marrow occurred. PMID- 6270973 TI - Temporary ischaemia induced by degradable starch microspheres. Possible thrombogenic effects in vivo and in vitro. AB - Possible thrombogenic effects of degradable starch microspheres were investigated. Controlled temporary small intestinal ischaemia ws induced by injection into the superior mesenteric artery in cats. Arterial flow consistently recovered after ischaemia. No consumption of blood platelets, fibrinogen, or Factor VIII was observed. Aggregation of human platelets was not influenced by microsphere exposure, and platelet retention in starch microsphere columns was minimal. No thrombosis was detected in feline small intestinal vessels in vivo nor did starch surfaces induce adhesion or aggregation of human platelets in vitro. Thus, no evidence of thrombotic hazards was found by inducing temporary intestinal ischaemia by starch microspheres. PMID- 6270974 TI - Tocopherol in irradiation of experimental neoplasms. Influence of dose and administration. AB - The influence of tocopherol, administered intramuscularly in different single doses, on the effect of local irradiation of a transplanted rat sarcoma was investigated. Tocopherol in doses of 5, 25 or 50 mg/100 g body weight enhanced significantly the tumour growth retardation induced by irradiation. Tocopherol in a dose of 100 mg/100 g body weight as a placebo preparation had no similar effect. The influence of tocopherol administered intramuscularly or orally in identical doses was similar. PMID- 6270975 TI - Cure rate after 131I therapy for hyperthyroidism. AB - The cure rate of hyperthyroidism after 131I therapy was analysed in 4 473 patients, aged between 14 and 91 years, treated between 1951 and 1975. Despite unchanged treatment principles, the cumulative incidence of cure increased for each 5-year period. Within the first 4 months 34 per cent of the patients with diffuse thyroid glands treated during 1951-1955 were cured, as compared with 59 per cent of those treated 1971-1975, i.e. an increment by a factor of 1.7. The corresponding figures for patients with nodular thyroid glands were 23 and 54 per cent, respectively, or a factor of 2.3. PMID- 6270976 TI - Radiation therapy in the management of malignant thymic tumors. AB - A review of 20 patients with malignant thymic tumors is presented. A simplified classification is used avoiding the term thymoma, which is considered misleading. Most of the patients were treated by a combination of surgery and radiation therapy while 7 were treated by irradiation alone. Postoperative radiation therapy is indicated when the tumor is incompletely removed. This is also advised in invasive tumors even when they are considered completely resected. Irradiation alone of inoperable tumors should be carried out with high doses (55 to 60 Gy) to decrease the risk of local recurrence. PMID- 6270977 TI - Clinico-pathologic correlation in non-Hodgkin's lymphoma. IV. Analysis of patients with clinically localized disease. AB - A retrospective analysis of 140 patients with non-Hodgkin's lymphoma in clinical stage I or II classified according to a modified LUKES & COLLINS scheme was performed. Three major groups were found according to cell type, with different clinical features: (1) Small cell lymphomas with a relatively favourable survival in spite of high relapse rates. (2) Large cell lymphomas with lower relapse rates, but short time between relapse and death, and unfavourable survival. (3) Mixed small/large cleaved follicular centre cell lymphoma which was most favourable with respect to relapse and survival. Nodular lymphoma had the same overall relapse rate as diffuse lymphoma, but had a significantly longer survival. Tumours stage I were associated with significantly longer relapse-free survival and survival than stage II. The importance of separating the majority of non-Hodgkin's lymphomas into three main groups according to cell type is emphasized. These major groups require different clinical approaches in terms of staging and treatment. PMID- 6270978 TI - Role of bone scans in the initial assessment of operable patients with breast carcinoma. AB - The yield and the prognostic significance of initially performed bone scintigraphy in 387 patients with operable mammary carcinoma are reported. The mean follow-up time was 52 months. In 12 patients (3.1%) the scintigraphy was considered to indicate bone metastases but in only 3 of these bone metastases later developed. The low information obtained has resulted in exclusion of bone scintigraphy in the initial assessment of clinically operable patients with mammary carcinoma. PMID- 6270979 TI - 14C leucine uptake in rat tissues at different times after irradiation. AB - The uptake of 14C leucine administered at different intervals after irradiation, but always 4 and 8 h before the animals were killed, has been evaluated in tissues with different proliferative activity and protein synthesis. The results have demonstrated an increased uptake and a more rapid elimination of the tracer after irradiation. In the small intestine a lower amount of TCA insoluble fraction was observed when the morphologic injury was evident, while protein synthesis significantly increased during the initial phase of appearance of the injury and mainly during the recovery phase of epithelial cells. Kidney and plasma had levels higher than controls at all intervals. PMID- 6270980 TI - Comparison between two radiation protective substances in irradiated mice. Effect on splenic haemopoiesis. AB - The radiation protective action of 2,2'-Dithiobis(N-[(1-adamantyl)-methyl] acetamidine)-dihyrochloride (S-75) and cysteamine was compared in splenectomized and non-splenectomized mice. Cysteamine was found to have better and more general protection properties. Several indications of a specific effect of S-75 on the spleen were observed. It is suggested that the protection properties of S-75 should be tested in another laboratory animal not having such a marked splenic haemopoiesis as the mouse. PMID- 6270981 TI - Microcephaly, mental retardation and chromosomal aberrations in a girl following radiation therapy during late fetal life. AB - A human foetus was heavily irradiated in the thirtieth to the thirty-third week due to carcinoma of the uterine cervix of the mother. Irradiation after 20 weeks of pregnancy is thought not to produce severe abnormalities. However, the child showed microcephaly, mental retardation, stunted growth, microphthalmus, retinal degeneration, cataract and defective dentition. Cytogenetically the frequencies of both chromatid and chromosome breaks were increased. PMID- 6270982 TI - Quality control in mammary radiography. AB - The doses, the image qualities and the factors affecting these were determined for all equipments used for mammary radiography in Finland. It was found that the quality of the equipment tested varied greatly. This is particularly well illustrated by the 5 000-fold difference between the highest and lowest surface doses. Large differences in image quality were also found. PMID- 6270983 TI - Generation of superoxide and hydrogen peroxide during interaction of nitrite with human hemoglobin. AB - Generation of superoxide and hydrogen peroxide during interaction of nitrite with human hemoglobin was detected by chemiluminescence of luminol. Luminol chemiluminescence was inhibited by the addition of superoxide dismutase (SOD) and catalase. Methemoglobin formation induced by nitrite was also inhibited by the addition of SOD and catalase. The mechanism of methemoglobin formation by nitrite was discussed in regard to the oxidation of hemoglobin by superoxide and hydrogen peroxide as generated by the interaction of nitrite with hemoglobin. PMID- 6270984 TI - Blood monocytes and serum and bone marrow lysozyme in sarcoidosis. AB - Serum lysozyme (LZM) concentrations were correlated to the number of neutrophils and monocytes in patients with sarcoidosis and nongranulomatous diseases. In sarcoidosis patients with an increased activity of serum angiotensin converting enzyme (ACE), a positive correlation was noted between LZM and blood monocytes. In sarcoidosis patients with normal ACE activity, as well as in patients with non granulomatous diseases, a correlation was found between blood neutrophils and LZM, but not between blood monocytes and LZM. LZM was found in bone marrow plasma and in serum in a ratio of 1.5 to 1. Sarcoidosis patients had 30% higher LZM levels than healthy controls. The concentration of LZM in bone marrow plasma did not correlate to detectable granulomas in bone marrow specimens. The positive correlation between blood monocytes and LZM in patients with clinically active sarcoidosis is possibly due to recruitment of bone marrow monocytes for the granuloma formation. PMID- 6270985 TI - Correlation between disease activity, one-year prognosis, and angiotensin converting enzyme in untreated sarcoidosis. AB - The correlation between S-angiotensin-converting enzyme (SACE) and disease activity was examined in 185 observation periods in 85 untreated sarcoidosis patients. An agreement between SACE and chest roentgenographic changes was found in 42% of the observations, although less frequently in patients with erythema nodosum. The most convenient interval between enzyme measurements seems to be 3 months in patients with active disease, but more frequent check-ups are advised in patients with erythema nodosum. In 24 patients followed for one year after diagnosis, SACE correlated fairly well to the clinical course, and the pattern of enzyme variation may be a prognostic indicator in sarcoidosis. PMID- 6270986 TI - Influence of calcium infusion on urinary cyclic AMP and phosphate in hyperparathyroidism. AB - Urinary cyclic AMP (cAMP) and phosphate were measured before and after calcium infusion in 12 patients with operatively verified primary hyperparathyroidism (PHP) and in 12 healthy persons. In normal subjects infusion of calcium caused a reduction in urinary cAMP directly correlated to the preinfusion values and inversely correlated to the serum calcium concentration determined as albumin corrected serum calcium. In normal subjects with high normal albumin-corrected serum calcium the infusion of calcium caused no or only a small depression in the urinary excretion of cAMP. Changes in phosphate excretion were not correlated to the calcium concentration. Four of the 12 hyperparathyroid patients showed normal relative suppression in urinary cAMP after the infusion of calcium, and 5 had normal suppression of phosphate excretion. It is concluded that some patients with PHP retain calcium-sensitive secretion of PTH, and that the classical calcium infusion test is of doubtful value in the diagnosis of PHP. PMID- 6270987 TI - Update on pancreatic polypeptide as a specific marker for endocrine tumours of the pancreas and gut. AB - Serum levels of pancreatic polypeptide (PP) were determined in 31 patients with endocrine tumours, localized in the pancreas (n = 16), small intestine (n = 13) and respiratory tract (n = 2). Two further patients with laxative abuse were investigated. Elevated serum levels in the peripheral circulation were noted in 56% of the patients with pancreatic tumours, in 60% of those with gut or bronchial tumours and in both patients with laxative abuse. Our study suggests that the elevated PP concentrations found in connection with endocrine gastrointestinal tumours originate from non-tumour pancreatic PP cells and only occasionally from the tumour. PP is a valuable tumour marker for endocrine gastrointestinal tumours, although it is not specific for pancreatic tumours. Certain inflammatory diseases, renal function and consumption of laxatives must be considered when evaluating elevated PP levels. Serum PP seems to be of limited value for evaluating the response to cytotoxic therapy. PMID- 6270988 TI - HCG-alpha and HCG-beta subunits as tumour markers during therapy in a case with so-called "non-functioning" islet cell tumour. AB - A 61-year-old man with a malignant endocrine pancreatic tumour, so-called "non functioning" islet cell tumour, is described. The tumour consisted of enterochromaffin-like cells with positive immunocytochemistry for gastrin, glucagon and VIP, but neither of these or other peptides were elevated in the circulation. Elevated serum levels of HCG-alpha and HCG-beta subunits were found. They seemed to be valuable tumour markers during cytotoxic therapy. PMID- 6270989 TI - Scintigraphic assessment of heterotopic cardiac transplants. AB - Patients receiving heterotopic ("piggyback") cardiac transplants, when the patient's own and transplanted donor hearts are connected in parallel, present special problems in determining their relative contributions to total cardiac function. Three patients who had transplants because of intractable heart failure were studied using first pass and gated equilibrium technetium-99m-labeled blood pool scintigraphy. In one patient, thallium-201 myocardial perfusion scans were obtained. These nuclear cardiology techniques provided anatomic and functional information noninvasively that proved helpful in patient management. PMID- 6270990 TI - CT of soft-tissue tumors. AB - Computed tomography was performed on 52 patients with soft-tissue tumors, 36 malignant and 16 benign. Major advantages with CT include the possibility of differentiating between lipoma and other tumors and the ability to demonstrate the transverse location of a lesion (intra- or extracompartmental). It was not possible to relate attenuation values to histologic type or grade of malignancy. CT was found superior to conventional radiography in demonstrating additional bony destruction of the pelvis and spine. The risk of overestimating the size of high grade malignant tumors because of accompanying edema is discussed. CT should precede angiography in the investigation of soft-tissue tumors, and angiography may primarily be reserved for those lesions where vascular relationships are not adequately demonstrated by CT. PMID- 6270991 TI - Infarction of renal tumors using isobutyl-2 cyanoacrylate and lipiodol. AB - Percutaneous transcatheter embolization of the renal artery in clear cell carcinoma was performed in nine patients. The occlusive material used was a suspension of 1 ml of isobutyl-2-cyanoacrylate in 3 ml of Lipiodol. Six patients were operated on 2-5 days after the embolization, and in three patients the procedure was palliative. In one of the latter, follow-up arteriography 4 months later showed that the artery was still occluded. There were no complications attributed to the glue or to oil emboli in this series. PMID- 6270992 TI - Follow-up of Wilms tumor: comparison of CT with other imaging procedures. AB - In a retrospective review, computed tomography (CT) was compared to a "routine" combination of other diagnostic imaging procedures used for follow-up evaluations of 13 children being treated for Wilms tumor. The examined variables were diagnostic accuracy, expense, and duration of examination. Results from 13 patients indicated that CT most accurately answers diagnostic queries pertinent to follow-up evaluation of Wilms tumor: the presence and extent of bilateral renal tumors, local recurrence, contralateral renal hypertrophy, and metastasis to liver or lungs, For diagnosing pulmonary metastases, CT was superior to conventional chest radiography both in sensitivity (4/4 vs. 2/4) and specificity (9/9 vs. 6/9). In depiction of liver metastases, CT (3/3) was superior to liver scintigraphy (2/3). The extent of bilateral Wilms tumors was better defined by CT than by urography. In no instances were the alternative diagnostic imaging studies found to be more accurate than CT for detection of recurrent tumor. Average cost for a CT examination ($344) is considerably less than the cost for a routine combination of the other imaging studies ($594). Examination time and diagnostic radiation does are also reduced using CT. Pending larger comparison studies, CT is recommended as the primary diagnostic method for follow-up evaluation of patients with Wilms tumor. PMID- 6270993 TI - Perfusion scan in pulmonary vascular/lymphangitic carcinomatosis: the segmental contour pattern. PMID- 6270994 TI - Cleansing the colon in gallium-67 scintigraphy: a prospective comparison of regimens. AB - Colonic accumulation of gallium-67 frequently complicates the interpretation of gallium-67 scintigrams. Although various modes of cleansing the colon prior to scintigraphy have been suggested, there is controversy over their efficacy and none have been tested prospectively. Three hundred nine patients undergoing gallium-67 scintigraphy were randomly assigned to one of four cleansing regimens: (1) a high fiber diet (78 patients); (2) castor oil (76); (3) milk of magnesia and cascara (76); and (4) not preparation (79). Patient compliance rates for the four regimens were 17%, 32%, 36%, and 46%, respectively. After noncompliant patients were excluded, gallium-67 scintigrams were graded for colonic activity on a scale of 0-3 by three independent, experienced observers. Gallium-67 activity in the colon was significantly less after administration of castor oil than after no preparation (p = 0.047). A high fiber diet also resulted in a substantial reduction of colonic activity when compared with no preparation; the difference, however, was not statistically significant (p = 0.083). Regimen 3 did not produce significantly better results than regimen 4 (p = 0.42). A major impediment to the success of any cleansing regimen seems to be poor compliance of patients. PMID- 6270995 TI - Immunocomplexes and primary lung cancer. AB - Circulating immune complexes (CIC) were found in 47% of 71 patients before treatment, in a horizontal study of the presence of CIC in primary lung cancer (PLC). The precipitation technique in PEG was used, and C1q, IgG and IgM fractions in the precipitate were assayed. Especially in epidermoid carcinoma, CIC were found in 1/6 cases with stage 1, 2/9 with stage II and 21/34 with stage III. The frequency was significantly higher in stage III cases, and showed a significant tendency to increase with progression of the clinical stage. If stage parameters are considered separately, this tendency correlates significantly (p less than 0.002) with the amount of lymph node involvement (N) but not to the tumor (T) size nor to the presence of metastases (M). Analysis of CIC behavior in comparison with the single fractions showed that C1q and/or IgG were elevated in 40% of cases. The presence of C1q and/or IgG associated with IgM is more characteristic of stage N2, while the presence of CIC with the fractions C1q and/or IgG not associated with IgM is more frequent in stage N1. There was a lower survival rate at 12 months in patients with CIC as compared to those without (p less than 0.05). PMID- 6270996 TI - Blood pressure and diet in normotensive volunteers: absence of an effect of dietary fiber, protein, or fat. AB - In the course of four controlled experiments on the effect of specific dietary components on cardiovascular risk factors, the effects on blood pressure of various sources of dietary fiber, of type and amount of dietary fat, and of animal versus plant were measured in young normotensive volunteers. In each of the four experiments a group of 50 to 75 healthy student volunteers received a control diet for 11/2 to 21/2 wk. They were then randomized into subgroups which received various test diets for periods ranging from 4 to 12 wk. In each experimental one group received the control diet throughout the whole experimental period. Diets differed between groups in one dietary component only. All foodstuffs were weighed out individually according to each person's energy needs. Body weights and Na intake were controlled. Initial blood pressures were about 120 mm Hg systolic and 70 mm Hg diastolic. Both systolic and diastolic blood pressure decreased during the test period in all four experiments on almost every diet, including the control diets, by about 0 to 5 mm Hg. However, changes in blood pressure over the test period were never significantly different between the test groups and the control groups. Thus, none of the investigated dietary factors had a demonstrable effect on blood pressure in young normotensive persons. PMID- 6270997 TI - Predicting weight loss success among obese clients in a hospital nutrition clinic. AB - A retrospective study of 63 obese patients seen in a hospital outpatient nutrition clinic was reported. The majority suffered from one or more other medical problems. The relationship of amount of and rate of weight loss to various factors was examined. The multivariate analysis revealed that either the amount or rate of weight loss was greater in the patients who were more overweight initially, white, male, young, single, older at age of onset of obesity, visited the clinic for a longer period of time but less frequently. Conversely, the amount or rate of weight loss was less in those patients receiving conjugated estrogens, oral contraceptives, propranolol, thioridazine HCl, and those who were receiving behavior modification as treatment. PMID- 6270998 TI - Alterations of fecal steroid composition induced by changes in dietary fiber consumption. AB - The short-term effects of high carbohydrate diets of normal foods either high or low in dietary fiber on fecal steroids and fiber was assessed in eight healthy young men. Each subject consumed in random order for 4 days a diet containing 59 g (high fiber) or 21 g (low fiber) neutral detergent fiber. After a 9-day rest period he consumed the other diet. Analysis of random fecal samples during their usual diet and after 4 days of each experimental diet showed an increased in primary bile acids from less than 4 to 32% of total bile acids, and a decreases of coprostanol from 76% (control diet) or 64% (low fiber diet) to 45% of total neutral sterol after the high fiber diet. Fecal fiber concentration doubled after the high fiber diet. We conclude that 4 days of high fiber diet is sufficient to cause a large increase in primary and decrease in secondary fecal steroids. Such changes have implications for prevention of arteriosclerosis and cancer of the colon. PMID- 6270999 TI - The effect of rolled oats on blood lipids and fecal steroid excretion in man. AB - Rolled oats (125 g daily) were substituted for breakfast cereals and wheat flour in the metabolically controlled diets of 10 subjects for 3 wk. Fat and energy intakes in the 2-wk control periods before and after the oat period were adjusted by addition of an oil with a similar fatty acid composition to the lipid in the oats. Plasma total cholesterol concentrations were reduced in seven of 10 subjects, but over the whole group the mean reduction of 8% was not significant (0.05 less than p less than 0.01). High-density lipoprotein cholesterol concentrations and plasma triglyceride levels were unchanged. Fecal fat excretion was increased by 47% (p less 0.005) and fecal bile acid excretion by 35% (p less than 0.01) but neutral steroid excretion was unchanged on the oatmeal diet. PMID- 6271000 TI - The effect of dietary citrus pectin on the excretion of human fecal neutral and acid steroids and the activity of 7alpha-dehydroxylase and beta-glucuronidase. AB - The purpose of this study was to investigate the effects of citrus pectin on human fecal neutral and acid steroid excretion and beta-glucuronidase and 7alpha dehydroxylase activity. Eight healthy male subjects (age 20 to 27 yr) were used in a switchback design with or without 15 g citrus pectin added to a mixed low fiber diet. There were three successive 18-day periods preceded by a 4-day adjustment period. Half of the subjects followed a pectin-nonpectin-pectin protocol and the other half followed a nonpectin-pectin-nonpectin protocol. Fecal samples were collected throughout the study under anaerobic conditions. Compared to the control diet, mean fecal weight, percentage moisture, transit time, and fecal fat for both groups of subjects were not significantly different by analysis of variance when subjects were fed pectin diet. Mean beta-glucuronidase activity was increased (35%) when subjects were fed the pectin. Mean 7alpha dehydroxylase activity showed no definite trend. Mean neutral steroid concentration was slightly decreased (8%) when the pectin diet was fed but total excretion was unchanged. End of period neutral steroid concentration was decreased 9% and total excretion was decreased 3.5%. Mean acid steroid concentration was not changed but total excretion was increased (11%) on the pectin diet. End of period acid steroid concentration and excretion was increased 6% on the pectin diet. This study shows that there were interrelationships between dietary pectin, neutral and acid steroid metabolism, and bacterial enzyme activity. PMID- 6271001 TI - Fecal bulk, energy intake, and serum cholesterol: regression response of serum cholesterol to apparent digestibility of dry matter and suboptimal energy intake in rats on fiber-fat diet. AB - Two experiments were conducted in the rat to determine the relationships of serum cholesterol (SC, mg/dl), apparent digestibility of dry matter (DDM, %), and digested energy intake (DE, kcal/day) at suboptimal level of energy. The energies in diet and feces were determined by calorimetry. DE as percentage of the National Research Council requirement (DE%) was suboptimal (70 to 85%). The experiments had four to five isofibrous diets, and no fiber diets, supplemented with 0.2% crystalline cholesterol (CChol). Animals in experiment 1 were fed varying amounts of feed with 18% coconut oil in the diets where as these in experiment 2 were given fixed amounts of feed with either 6 or 18% oil. The following regressions (p less than 0.001) for SC were found: experiment 1: 1157.7 -5.97 DDM +105.5 CCI -1.48 CCI2 (r2 0.35), where CCI = CChol, mg/day; 1888.4 -2.66 DE +120.97 CCI -1.62 CCI2 (r2 0.37). Experiment 2: 762.99 -6.15 DDM 0.8 fat cal % -0.87DE% (r2 0.31), where fat cal % = fat calories % of DE. Data indicate that at suboptimal energy intake, SC was inversely related to (1) DDM, (2) fat cal, and (3) total energy intake. Liver cholesterol lowering effect of the dietary fiber was also observed. The above findings help to elucidate various conflicting reports related to diet and blood cholesterol. PMID- 6271003 TI - Influence of dietary iron deficiency on hemoglobin, myoglobin, their respective reductases, and skeletal muscle mitochondrial respiration. AB - Male weanling rats were fed a control diet (46 ppm iron) or an iron-deficient diet (11 ppm iron) for 7 wk to determine the influence of iron deficiency on heme proteins and skeletal muscle mitochondrial respiration. At the end of 7 wk, the hemoglobin in the blood of the iron deficient rats was 35% less and skeletal muscle myoglobin was 20 to 37% less than in the control animals. The concentration of myoglobin in the heart was not appreciably diminished by iron deficiency. Cytochrome c concentration was 20% less in the heart and 35% less in the mixed-fiber gastrocnemius in the iron-deficient animals. Iron deficiency did not influence the activity of metmyoglobin reductase in either heart or skeletal muscle. There was about 30% more methemoglobin reductase activity in the red blood cells of the iron-deficient animals, which resulted in methemoglobin levels that were so low as to be virtually unmeasurable. In the iron-deficient rats, skeletal muscle mitochondrial respiration with either pyruvate-malate or palmitylcarnitine as substrate was 17 to 20% less than in the control animals. This study demonstrates that dietary iron deficiency of sufficient severity to reduce blood Hb and skeletal muscle myoglobin or cytochrome c also results in an impaired skeletal muscle oxidative capacity. The study also illustrates the preferential utilization of iron, not only between tissues, but within tissues, and tissue specific adaptive responses to iron deficiency. PMID- 6271002 TI - Limits of predicting gastrointestinal transit time from other measures of bowel function. AB - The two aims of these experiments were first, to examine the relationships between mean daily stool weight, mean weight per stool, frequency of defecation, dye transit time, dietary fiber intake, and transit time of small radiopaque pellets, and second, to determine the ability of these measures of bowel function, singly or in combination, to predict gastrointestinal transit time of the pellets. Variables were observed simultaneously in 13 healthy women consuming controlled low and high cellulose diets. All except one of the correlation coefficients between the measures of bowel function were significant. Stepwise regression analysis retained log stool weight, fiber and dye transit to predict log of the mean transit time of all pellets (R2 = 71.3); the same three variables were selected to predict log of the transit time of 80% of the pellets (R2 = 59.4). Mean daily stool weight explained about 50% of the variation for both measures of transit, while dye transit and dietary fiber explained the rest. Log transformation of either stool weight or pellet transit time or both variables improved the prediction of about 10%. These results suggest that prediction of pellet gastrointestinal transit time from other measures of bowel function may be limited and is influenced significantly by the fiber level of the diet. PMID- 6271004 TI - Detection of herpes simplex virus (HSV) type-1 IgG and IgM antibodies by enzyme linked immunosorbent assay (ELISA). AB - A sensitive enzyme linked immunosorbent assay (ELISA) is described for detection of herpes simplex virus (HSV) type 1, IgG and IgM antibodies. The antigen consisted of a crude extract of HSV infected human foreskin fibroblast cells. Specific horseradish peroxidase conjugated antisera were used to detect total immunoglobulin, IgG and IgM antibodies bound to viral antigen. The substrate was a solution of 5-aminosalicylic acid and hydrogen peroxide, which yielded a readily visible endpoint. Results obtained by the ELISA method were compared with the micro-complement fixation (CF) method on 36 sera. ELISA was shown to be at least 10-20 fold more sensitive than CF, with a correlation coefficient of 0.752 (p less than .001). HSV antibodies in these sera were mainly IgG, although IgM antibodies could also be detected by ELISA. HSV antibodies were not found in 16 cerebrospinal fluids from patients without HSV encephalitis (HSVE). ELISA appears to be a rapid, sensitive, and specific method for demonstration of IgG and IgM HSV antibodies. It may have possible application in the diagnosis of HSVE. PMID- 6271005 TI - IgA nephropathy associated with bronchial small-cell carcinoma. AB - Two cases of patients with a bronchial small-cell carcinoma coexisting with IgA nephropathy are reported. The relationship of the bronchial malignancy to IgA nephropathy is discussed. PMID- 6271006 TI - Pathogenetic aspects of hepatitis A virus infection in enterally inoculated marmosets. AB - Experimental hepatitis A virus (HAV) infection was studied in marmosets after enteral (intragastric) inoculation with special reference to the primary sites of HAV replication and immunopathology of the disease. The experiment was carried out using 28 Saguinus mystax negative for antibody to HAV (anti-HAV) and with statistically uniform baseline values of serum isocitrate dehydrogenase (SICD) activity. Each animal was infected with 1 ml of a 15% w/v stool suspension that was derived from marmosets infected with the third or fourth passage of the MS-1 strain of HAV. The incubation period measured by the first significant SICD elevation was 32 days in 11 of 13 marmosets. The animals were sacrificed 2, 4, 7, 11, 14, 18, 23, 28, and 32 days after inoculation and 1, 4, 8, 14, 21, 28, and 35 days after SICD elevation. HAV antigen, immunoglobulins, complement, and fibrin were identified in the liver, eight segments of the gastrointestinal tract, lymphoid system, and kidneys. HAV antigen was found only in the cytoplasm of hepatocytes and in gallbladder bile. These findings indicated that the liver was the sole and primary site of virus replication. Combined immunomorphologic and histopathologic observations also revealed that HAV antigen localization was associated with the sites of hepatocellular damage. There was no immunomorphologic evidence for humoral immune clearance of HAV antigen in the liver, lymphoid system, or kidneys. PMID- 6271007 TI - Varicella. PMID- 6271008 TI - Vitamin D metabolism in idiopathic infantile hypercalcemia. AB - A 9-month-old boy who had the mild form of idiopathic infantile hypercalcemia was observed for 18 months. During the initial hypercalcemic stage, the serum concentration of 25-hydroxyvitamin D was normal. Urinary levels of cyclic adenosine monophosphate (cAMP) were low, and the serum concentrations of the dihydroxyl metabolites of vitamin D were appropriate to the high serum calcium concentration, with low 1,25-dihydroxyvitamin D and relatively high 24,25- and 25,26-dihydroxyvitamin D levels. Throughout the study period, there was a close positive correlation between the magnitude of the urinary cAMP excretion and the serum level of 1,25-dihydroxyvitamin D. The results indicate that excessive vitamin D intake leading to high serum levels of 25-hydroxyvitamin D are not decisive factors in the pathogenesis of idiopathic infantile hypercalcemia. PMID- 6271009 TI - Vitamin D metabolite concentrations in vitamin D deficiency. Are calcitriol levels normal. AB - The levels of vitamin D metabolites were measured in three children with a decreased dietary intake of calcium and vitamin D and sun exposure. All three children had hypocalcemia, hypophosphatemia, and elevated alkaline phosphatase activities. Two children had rickets, aminoaciduria, and elevated immunoreactive parathyroid hormone (iPTH) concentrations. The concentrations of vitamins D2 and D3, 25-hydroxyvitamins D2 and D3 (25-OH-D2 and 25-OH-D3), and 24,25 dihydroxyvitamin D (24,25-[OH]2D) were reduced. Nonetheless, the levels of calcitriol (1,25-[OH]2D) were normal. The combination of hypocalcemia, hypophosphatemia, and increased iPTH concentrations should result in supranormal calcitriol concentrations. Moreover, the ratio of PTH to calcitriol is significantly higher than in normal subjects. Accordingly, in patients with vitamin D deficiency and "normal" calcitriol values, the synthesis of this compound may be reduced. The evaluation of vitamin D deficiency should include the measurement of all metabolites. PMID- 6271012 TI - Stevens-Johnson syndrome associated with adenovirus conjunctivitis. PMID- 6271010 TI - Transient adrenogenital syndrome due to exposure to danazol in utero. AB - We describe a premature female infant exposed in utero to danazol during the first trimester of pregnancy. She was first observed in the newborn period with marked degree virilization and clinical findings suggestive of salt-losing congenital adrenal hyperplasia. This was supported by the high plasma levels of 17 alpha-hydroxyprogesterone and adrenocorticotropic hormone and low plasma cortisol level. Levels of testosterone, androstenedione, 11-deoxycortisol, and renin were also elevated. An excessive increase in the levels of 17 alpha hydroxyprogesterone and 11-deoxycortisol to corticotropin administration associated with impaired increase in plasma cortisol level strongly suggests a partial block in the 21-hydroxylation of 17 alpha-hydroxyprogesterone. However, the high levels of 11-deoxycortisol also suggest a block of the steroid 11 beta monooxygenase. A year later she was found to have normal basal levels of the adrenal steroids and normal response to corticotropin administration, pointing out the transitory nature of these abnormalities. It may be hypothesized that danazol produced a transitory block of the steroid 21- and 11 beta-monooxygenases in this child. PMID- 6271011 TI - Are female sex hormones teratogenic? AB - An analysis of available epidemiologic data leads the present reviewers to conclude that the use of exogenous hormones during human pregnancy has not been proved to cause developmental abnormality in nongenital organs and tissues. This conclusion is further supported by the animal laboratory data. The preponderance of evidence at this writing indicates a lack of causal association between hormonal use during pregnancy and nongenital malformation of the offspring. The quality of the epidemiologic data does not, at this time, permit a definitive conclusion that sex hormones during pregnancy may not, under as yet to be defined conditions, have some adverse effect on human prenatal development. If there are increased risks of nongenital malformations associated with the administration of certain sex steroids, the risks are very small, may not be causal, and are substantially below the spontaneous risk of malformations. In spite of the present degree of uncertainty, the clinical, epidemiologic, and laboratory data do permit the formulation of a rational approach to handling problems related to sex steroid usage and exposure in pregnant women. PMID- 6271013 TI - Immunopathology of natural and experimental lymphomas induced by wild mouse leukemia virus. AB - Naturally occurring lymphomas of Lake Casitas (LC) wild mice, and the lymphomas induced by LC murine leukemia virus (MuLV) in Swiss mice from the National Institutes of Health, displayed remarkably similar gross, microscopic, and functional characteristics. They spared the thymus, arose primarily in the splenic red pulp, became leukemic, and were comprised of stem cells lacking classic T- and B-cell markers. Cytoplasmic and surface immunoglobulin were undetectable in 34 of 35 spontaneous LC lymphomas and in any of ten LC MuLV induced lymphomas in NIH Swiss mice. Assays for immunoglobulin secretion, complement (C'3) and Fc receptors, Thy 1.1,2 antigens, Ly 1,2 antigens, and erythroid and myeloid markers were negative on all of the spontaneous and experimental lymphomas. Cell lines were derived from five spontaneous lymphomas of LC mice. Three lines were characterized as null cells, one line as B cells, and one line as macrophages. All cell lines were diploid. The wild mouse spontaneous lymphomas, and lymphomas experimentally induced by LC MuLV in laboratory mice, provide a useful model for childhood acute lymphoblastic leukemia and for study of the early steps of B-lymphocyte differentiation. PMID- 6271014 TI - Secretory component: a glandular epithelial cell marker. AB - Secretory component (SC) has been demonstrated to be produced by both normal and malignantly transformed glandular epithelial cells. By an indirect immunofluorescent technique, this study surveys tumors of varied cellular origin in order to determine the reliability of SC as a marker for tumor cells derived from glandular epithelium. Both primary and metastatic tumors of glandular epithelial origin demonstrated SC fluorescence, while nonglandular epithelial tumors did not. This observation was extended to live single-cell preparations, which demonstrated intense cell-surface fluorescence only when glandular epithelial tumors cells were examined. Additionally, fixed, cytocentrifuged, single-cell preparations of glandular epithelial tumors demonstrated cytoplasmic SC fluorescence. When breast carcinoma was examined, all cases demonstrated SC, regardless of the degree of differentiation. This assay appears to have useful clinical application in that the finding of SC provides indication of the glandular epithelial origin of a malignantly transformed cell. PMID- 6271015 TI - Lung injury induced by antibody fragments to angiotensin-converting enzyme. AB - Rabbits given goat anti-rabbit angiotensin-converting enzyme antibodies or derived antibody fragments develop rapidly fatal pulmonary edema. Endothelial cell injury is manifested by bleb formation and the disintegration of cell membranes. Platelets are found along the injured endothelium and leukocytes block capillary lumens. The pathologic features are similar when immune IgG, F(ab')2, or Fab are given. In vitro studies of complement activation show that solubilized, purified angiotensin-converting enzyme alone activates C1, with consumption of C4 and C3. Addition of immune IgG plus converting enzyme enhances this activation. F(ab')2 plus enzyme enhances only C3 consumption, while Fab with enzyme produces no additional complement utilization. Thus, while complement activation may be involved in the pathogenesis of injury induced by IgG or F(ab')2, the mechanism of Fab-induced endothelial injury remains unclear. PMID- 6271017 TI - Sendai virus infection in genetically resistant and susceptible mice. AB - The pathogenesis of Sendai virus infection in genetically resistant (C57Bl/6) and susceptible (DBA/2) nonimmune adult mice was investigated. Rising serum complement-fixation (CF) antibody titers were delayed in DBA/2 mice as compared with C57Bl/6 mice. C57Bl/6 mice developed descending desquamative endobronchiolitis, and DBA/2 mice developed descending proliferative endobronchiolitis and bronchogenic alveolitis. Peribronchiolar lymphoid cuffs that formed in C57Bl/6 mice were thicker and more densely populated than those of DBA/2 mice. Immunofluorescence demonstrated viral antigens confined to the epithelial lining of conducting airways in C57Bl/6 mice but extending to alveolar corner cells in DBA/2 mice. Studies with a transmission electron microscope confirmed that Type II pneumocytes were infected only in DBA/2 mice. IgG containing cells selectively accumulated along the airways of both strains, but fewer were recruited by DBA/2 mice. These results suggest that genetic resistance to Sendai virus is expressed through the immune system. PMID- 6271016 TI - Role of extracellular calcium and neutrophil degranulation responses to 1-O-alkyl 2-O-acetyl-sn-glycero-3-phosphocholine. AB - The rabbit polymorphonuclear neutrophil degranulation response to 1-O-alkyl-2-O acetyl-sn-glycero-3-phosphocholine depends on extracellular calcium. In the absence of this bivalent cation, neutrophil suspensions pretreated with cytochalasin B responded to the lipid by releasing minimal amounts of lysozyme and beta-glucuronidase. Incremental increases in extracellular calcium over a range of 20-200 microM led to increasing amounts of lipid-stimulated enzyme release. In contrast, extracellular magnesium neither supported nor enhanced the degranulation responses. Verapamil (25-200 microgram/ml), a calcium channel blocker, inhibited degranulation. Neutrophil suspensions exposed to the phosphocholine stimulus rapidly took up radiolabeled extracellular calcium. The kinetics of this calcium uptake were similar to the kinetics of enzyme release, and the amount of calcium taken up correlated closely with the amount of released lysozyme and beta-glucuronidase. Finally, in a dosage which blocked degranulation, verapamil inhibited calcium uptake. Thus, the rapid association of extracellular calcium with the neutrophil may mediate, at least in part, the degranulating actions of the phosphocholine stimulus. PMID- 6271019 TI - Endogenous opioid activity and beta-endorphin immunoreactivity in CSF of psychiatric patients and normal volunteers. AB - The authors measured total opioid activity by radioreceptor assay in the CSF of 41 normal subjects and 89 unmedicated psychiatric patients, including schizophrenic, schizoaffective, depressed, and manic diagnostic groups. Schizophrenic men had significantly lower levels of opioid activity than the normal men, although these levels did not significantly differ from levels of other male patients. The authors observed higher opioid activity during mania than during depression in paired samples for 4 manic-depressive patients. beta Endorphin immunoreactivity in a subsample of the same subjects was no different in the patient group than in the normal group, suggesting that the differences in CSF opioid activity between schizophrenic men and normal patients may be related to opioids other than beta-endorphin. PMID- 6271020 TI - [Hypophyseal-adrenal system function in normally proceeding pregnancy]. PMID- 6271018 TI - Herpesvirus infection enhances cholesterol and cholesteryl ester accumulation in cultured arterial smooth muscle cells. AB - In our previous experiments, atherosclerosis similar to that in humans was reproducibly induced in both normocholesterolemic and hypercholesterolemic specific-pathogen-free (SPF) chickens by infection with Marek's disease herpesvirus (MDV). In contrast, uninfected chickens fed either relatively cholesterol-poor or cholesterol-supplemented diets did not develop this arterial disease. In experiments reported here, the hypothesis that infection of arterial smooth muscle cells (SMCs) with MDV would enhance lipid accumulation in these cells was tested. The number of MDV-infected SMCs with lipid stained with oil red O was assessed, and the lipid content of these cells was quantitated chemically by chromatographic and fluorometric analyses. These data were compared to those of uninfected control cells and, in the case of chemical analyses, were also compared to SMCs infected with a second avian herpesvirus, turkey herpesvirus (HVT). Results indicate the following: 1) The percentage of MDV-infected SMCs containing stainable lipid was significantly greater than the percentage of uninfected SMCs; 2) Increased total lipid accumulation was observed in MDV infected SMC, particularly cholesterol (CH) and cholesteryl esters (CEs), as compared with uninfected or HVT-infected cells; 3) The types of CEs and nonesterified fatty acids (NEFA) accumulating in MDV-infected cells (particularly saturated types of CEs and NEFAs) were significantly different than those in uninfected or HVT-infected SMCs. These qualitative and quantitative differences in lipid content between infected and uninfected SMCs suggest that infection with MDV results in altered intracellular lipid metabolism. Results support the hypothesis that lipid accumulation in arteries of normocholesterolemic chickens may result from MDV infection acting at the cellular level to induce lipid accumulation that resembles that in human atheroarteriosclerosis. PMID- 6271021 TI - [Myometrial histochemical study of parturients with primary atomic labor]. PMID- 6271022 TI - [Possible participation of viral and mycoplasmal infections in inducing pregnancy complications]. PMID- 6271024 TI - [Burkitt's lymphoma]. PMID- 6271023 TI - [Haemodynamic effect of naloxone during electrostimulation analgesia with special reference to the role of endorphin (author's transl)]. AB - 10 persons undergoing cardiac surgery in electro-stimulation analgesia (ESA) with controlled ventilation were given 5 microgram/kg bodyweight and 10 microgram/kg body-weight respectively of naloxone during the operation and the effects of the drug on the circulation were registered over a period of 10 minutes. The observations do not support the view that endorphin plays a part in ESA. Other possible modes of action are discussed. PMID- 6271025 TI - Brain cyclic-AMP and possible mechanisms of cerebrovascular dilation by anesthetics in rats. AB - Barbiturates decrease both cerebral metabolic rate (CMR) and cerebral blood flow (CBF) and thereby preserve CBF and CMR coupling. However, many inhalation agents and ketamine uncouple CBF and CMR by virtue of inducing disproportionate increases in CBF. Adenosine 3',5'-monophosphate (cyclic-AMP) reportedly mediates uterine and aortic smooth muscle relaxation by halothane and may also mediate CBF CMR uncoupling by anesthetics. Therefore, the authors studied the effect of various doses of halothane and ether on brain cyclic-AMP and compared them with the effects of ketamine and pentobarbital anesthesia. Both halothane 1.5 and 2.0 per cent and ether, 2.5 to 7.5 per cent (inspired concentrations) increased whole brain cyclic-AMP above unanesthetized levels (1,046 +/- 75 pmol/g wet brain) in a dose-related manner. Ketamine, 150 mg/kg, ip, increased brain cyclic-AMP twofold, whereas an apparent increase by 60 mg/kg pentobarbital, ip, was not significant. These results show a dose-related effect of ether and halothane on brain cyclic AMP levels and suggest a causal relationship to their cerebrovasodilatory effects. PMID- 6271026 TI - Anesthesia for patients with insulinoma treatment with oral diazoxide. PMID- 6271027 TI - Respiratory distress and beta-endorphin-like immunoreactivity in humans. AB - Beta-endorphin-like immunoreactivity was determined in the plasma of twenty patients suffering from hypoxia of various etiologies and in twenty healthy adult volunteers who served as controls. Mean beta-endorphin-like immunoreactivity in the hypoxic patients was 53.2 +/- 5.5 (SEM) pg/ml, as compared to the volunteer subjects in whom the mean level was 6.2 +/- 1.9 pg/ml (P less than 0.01). Significant negative correlations were present between both arterial pH (r = 0.85; P less than 0.01) and arterial PO2 (r = -0.80; P less than 0.01) and beta endorphin-like immunoreactivity. These findings seem to lend support to the hypothesis that hypoxia and acidosis represent stressful conditions which may stimulate the release of beta-endorphin in humans. PMID- 6271028 TI - Introduction and reisolation of selected gram-positive bacteria from fermented edible wastes. AB - A fermentation process using Lactobacillus acidophilus added to edible food wastes was evaluated for its bactericidal action on selected gram-positive organisms. The Lactobacillus fermentation converts food wastes into an animal feed ingredient. In this study, 5 gram-positive bacteria of zoonotic importance were individually tested. These organisms were: Group E Streptococcus, Erysipelothrix rhusiopathiae, Clostridium perfringens, Corynebacterium pseudotuberculosis, and Listeria monocytogenes. For each experiment, Lactobacillus was first mixed into ground waste; one of the test organisms was then inoculated and mixed. This mixture was divided among eight 5.5-L containers and incubated (duplicates) at 5 C, 10 C, 20 C, and 30 C for 96 hours. Internal waste temperature, reduction-oxidation, and pH were monitored. Waste samples were taken initially and at subsequent 24-hour periods. Qualitative and quantitative recoveries of the test bacteria were attempted for each sample. Group E Streptococcus was reisolated in increasing numbers at all temperatures throughout the fermentation period. Erysipelothrix rhusiopathiae was recovered throughout the 96-hour period at 5 C; at 10 C it was recovered at 24 hours but not at 48 hours. Erysipelothrix rhusiopathiae was killed by 24 hours at 20 C and 30 C fermentation temperatures. Clostridium perfringens survived the entire test period at 5 C, 10 C, and 20 C; it was killed by 72 hours at 30 C. Neither Corynebacterium pseudotuberculosis nor Listeria monocytogenes was reisolated at any temperature at any time. PMID- 6271029 TI - Correlation of blood lymphocyte nuclear pocket prevalence with enzootic bovine leukemia p24 viral antibody titers. AB - The quantitative relationship between radioimmunoassay serotiters to p24 enzootic bovine leukemia viral antigen and the prevalence of lymphocytic nuclear pockets (LNP) was studied in 554 clinically normal dairy cows and bulls. Of the animals, 69% were seronegative. Of the seronegative animals, 44% had no observed LNP, whereas 93% of the seronegative animals had less than 1% LNP. However, 2.3% of the seronegative animals had substantially increased (greater than 2%) numbers of LNP, and conversely, 24% of the seropositive animals had no observed LNP. Statistical evaluation of the data, using contingency table and regression methods, provide significant (P less than 0.0001) evidence for a positive association between LNP prevalence and p24 bovine leukemia viral serotiters. Although significant, the association was weak, and hence LNP number had little value for predicting serotiter. PMID- 6271030 TI - Silicosis in silica flour workers. AB - In July 1979, the health of 86 current and ex-workers at 2 silica mining and milling operations in southern Illinois was examined using a respiratory questionnaire, spirometry, and chest radiographs. None of 25 current workers with less than 1 yr of exposure to silica dust had radiographic evidence of silicosis. For 61 current workers and ex-workers with 1 or more yr of exposure, chest radiographs showed 16 (26%) with simple silicosis and 7 (11%) with progressive massive fibrosis. Of these 23, 8 with simple silicosis and 3 with progressive massive fibrosis began work after the first Mine Safety and Health Administration inspection in 1973. These data and a review of federal dust inspection results between 1973 and 1979 showed that these cases of silicosis could have been prevented by effecting compliance with the existing dust standard. PMID- 6271031 TI - Electrodiagnosis in neuromuscular disorders. AB - Nerve conduction and stimulation studies and needle electromyography are useful in objectively verifying the presence of disease of nerve and muscle. How pathologic alterations in muscle and nerve produce the electrical abnormalities detected by these combined techniques is explored to guide the clinician in distinguishing whether a neuromuscular disorder involves the motor neuron, peripheral nerve axon or myelin, neuromuscular synapse, or muscle; whether the problem is focal or generalized; and whether there is evidence of ongoing disease or repair. PMID- 6271032 TI - Varicella-zoster virus infection among cancer patients. PMID- 6271033 TI - [Negativity of scintiscanning with technetium-99m-pertechnetate in cases of Meckel's diverticulum with heterotopic gastric mucosa]. PMID- 6271034 TI - [Angio-immunoblastic lymphadenopathy and neurological manifestations (author's transl)]. AB - Cases of associated angio-immunoblastic lymphadenopathy (AIL) and peripheral neuropathies have been rarely reported in the literature. Three such cases are described, in two men and one woman aged 79, 59, and 45 years respectively. Diagnosis of AIL was confirmed by lymph node biopsy in the two latter patients during their lifetime, and in the first case from examination of cervical nodes at autopsy. All three patients presented neurological disorders, of the polyradiculoneuritis type in the first case, multiple neuritis of the lower limbs followed by radiculalgia in the second, and myalgia and neuralgia with neurogenic signs in the EMG in the third case. No evidence of a toxic, metabolic, or infections aetiology was found, histological examination of nerve and muscle specimens demonstrated localised AIL lesions in one case, and discrete lymphoplasmocytic infiltration of the peripheral nervous system in the other two patients. PMID- 6271035 TI - [Polyneuropathy complicating Horton's disease (author's transl)]. AB - A patient diagnosed as having Horton's disease presented a complex neurological picture dominated by sensory-motor neuropathies of all four limbs, one year after the appearance of signs of temporal arteritis. No other etiological factor, apart from the Horton's disease, was discovered, and the causal relationship between this disease and the neuropathy is discussed. The possibility of dysimmunity factors being involved in Horton's disease is raised and the resulting therapeutic implications discussed. PMID- 6271036 TI - [Neurologic manifestations of Horton's giant cell arteritis]. PMID- 6271037 TI - [Isolation and characterization of a beta-lactamase-specifying plasmid in a strain of "neisseria gonorrhoeae" (author's transl)]. AB - GCL3, a beta-lactamase-producing, penicillin resistant strain of Neisseria gonorrhoeae isolated in Toulouse (France), and GCL51, a penicillin susceptible strain, were examined for their plasmid content. Agarose-gel electrophoresis following or not ultracentrifugation in a cesium chloride-ethidium bromide gradients, revealed, for both strains a 3.9-kilobase (kb) (2.6 megadaltons) cryptic plasmid. Penicillin resistant strain GCL3 also contained a 37 kb (24.5 megadaltons) and a 7.3 kb (4.8 megadaltons) plasmid. Transformation studies showed that the gene responsible for beta-lactamase production was carried by the 7.3-kb plasmid. E. coli cells transformed for ampicillin resistance by plasmid DNA from GCL3, contained a single 7.3 kb-plasmid. The restriction endonuclease cleavage map obtained for this plasmid indicated that it is closely related to a penicillin R plasmid previously described in a strain isolated in the USA. The first isolation in the Midi-Pyrenees area of a beta-lactamase-producing strain proved the necessity of a rigorous surveillance of N. gonorrhoeae strains isolated in the world. PMID- 6271038 TI - [Hormonal profile in anorexia nervosa under control. Correlations with various somatic factors]. AB - In order to precise the hormonal pattern of Anorexia Nervosa (AN) at its state phase, we assessed serum estradiol (E2), triiodothyronine (T3), LH and FSH, the maximum increments of their response to 100 microgram LHRH (delta LH and delta FSH) and the ratio of these increments (delta LH/delta FSH). These data were compared with those obtained in 10 normal women assessed at the early follicular phase (Student's test). Furthermore in 20 cases, we assessed LH and FSH during 5 days after a 20 mg IV injection of PREMARIN. E2, T3, LH and FSH are often decreased in AN. On the average, this decrease is very significant for the 3 first ones (p less than 0.001), and slightly significant for FSH (p less than 0,05). The LHRH-response is variable, with an inversion of the LH/FSH ratio as before the puberty in 56% of the cases. On the average, FSH-response is increased (p less than 0,05), and LH-response is normal. The response is delayed for the both gonadotropins. After PREMARIN, the 2 gonadotropins rarely clearly decrease (twice for LH and 5 times for FSH). There is a LH peak only in 3 cases. In order to specify the origin of these anomalies, we searched correlations between 11 hormonal data and some somatic parameters : weight, time, menstrual antecedents. We found 26 statistically significant correlations (12 times p less than 0,05- 5 times p less than 0,001- 9 times p less than 0,001). The following correlations are significant : E2 and LH, E2 and the maximal decrease of LH after PREMARIN (negative correlation), T3 and FSH, delta LH, and delta FSH, duration of the amenorrhea and T3, FSH and delta FSH, the weight and delta LH, the weight and delta LH/delta FSH, weight/height and delta LH/delta FSH, the menstrual antecedents and FSH. So, somatic factors strongly influence the hormonal pattern at the AN-state phase. The weight-decrease, the T3-decrease, and above all the E2 decrease which is independent of the previous ones seem to be the determining elements of the AN-hormonal pattern at this phase. PMID- 6271039 TI - Lysosomal abnormalities in cultured schwann cells from a patient with peripheral neuropathy and continuous muscle fiber activity. AB - Schwann cell cultures were established from a sural nerve containing large membrane-bound vacuoles in its Schwann cells, obtained from a patient with neuropathy and continuous muscle fiber activity. Cultured Schwann cells contained many large membrane-bound vacuoles, presumably lysosomes, resembling those present in the biopsied nerve. The acid phosphatase reaction was excessive in the patient's cultured Schwann cells but practically negative in normal cultured Schwann cells. This study indicates that the patient's neuropathy is primary dysschwannian with abnormal lysosomes as a major abnormality. PMID- 6271040 TI - Physiology and biochemistry of aerobic hydrogen-oxidizing bacteria. PMID- 6271041 TI - Evolutionary significance of accessory DNA elements in bacteria. PMID- 6271042 TI - Etiological implications on Kaposi's sarcoma. AB - In attempts to encompass multiple events responsible for the development of cancer in man, with a particular interest to the possible involvement of viruses, multidisciplinary studies have been conducted on KS during 1971-1979. There is accumulating evidence associating this malignancy with CMV. African KS patients have high antibody titers to CMV, but they are not significantly different from controls of the same regions (Ag-Ab complexes?). A specific serologic association of European and American KS with CMV was established. CMV-related antigens can be demonstrated in 22% of tumor biopsies and comparably in early passages of cell culture derived from them. Recently, CMV-DNA sequences have been detected in tumor biopsies. Furthermore, a CMV, strain K9V, has been isolated in tissue culture from a KS culture. Cells infected in vitro with K9V have shown some of the characteristics of cells transformed previously by other oncogenic CMV strains. In vivo K9V has induced a fatal disease in old world monkeys, mainly involving the lymphoid system. Moreover, a cluster type of occurrence of KS was detected in the West Nile District of Uganda, an area with as high incidence of KS as BL; and finally, there is a significantly higher incidence of a second primary tumor in KS patients, particularly of the lymphoreticular system. Whether CMV plays a role in the development of KS, a complex neoplasia which involves different cell types cannot be claimed by these results only. However, similarities between CMV and EBV, the latter being closely associated with BL and nasopharyngeal carcinoma are worthy of consideration. PMID- 6271043 TI - [Revision of the structure of ristomycinic and actinoidinic amino acids]. AB - The structures of ristomycin and actinoidine amino acids described earlier were revised. Crystalline derivatives of the amino acids and the products of their oxidation were prepared. The study on the spectral properties of the compounds showed that ristomycin and actinoidine amino acids had the structures of 3-(2' hydroxy-5'-glycyl-phenoxy)-4-methyl-5-hydroxyphenylglycine and 2-(2'-hydroxy-5' glycyl-phenyl)-3,5-dioxyphenylglycine respectively. They did not differ from deaminodicarboxylic acids prepared with ristocetin, vancomycin and actionoidine. PMID- 6271044 TI - [Response, to polymyxin, of plague microbe strains isolated from various natural foci and of their mutants with a decreased requirement in nutritional factors]. AB - Investigation of the response to polymyxin of 65 strains of Pasteurella isolated from various foci and 46 their back mutants showed that all of them were usually highly resistant to polymyxin (MIC 200--500 micrograms/ml). The Pasteurella strains isolated in the Gissaro-Darvaz natural focus, Turkey and Congo were highly sensitive to polymyxin (MIC 10--25 micrograms/ml). Single cultures highly sensitive to the antibiotic were detected among the polymyxin-resistant strains. Polymyxin-sensitive mutants of these cultures with lowered requirements in the growth factors were obtained. PMID- 6271045 TI - Comparative inhibition of influenza and parainfluenza virus replication by ribavirin in MDCK cells. AB - Actinomycin D inhibited the yield of influenza virus hemagglutinin from MDCK cells infected at high multiplicity, but had little effect on the yield of parainfluenza virus hemagglutinin. In similar hemagglutinin yield experiments, ribavirin was only slightly more active (threefold) against influenza virus than against parainfluenza virus replication. In plaque inhibition experiments, ribavirin depressed influenza virus plaque formation by 50% at a concentration of approximately 3 micrograms/ml, whereas the corresponding figure for parainfluenza viruses was threefold higher. The concentration of ribavirin demonstrating anti influenza activity was indistinguishable for that inhibiting host cell growth. It is concluded that, unlike actinomycin D, ribavirin is unlikely to have a major effect on the provision of host cell 5-germinal methylated cap structures and the subsequent priming of influenza messenger ribonucleic acid synthesis. PMID- 6271046 TI - Aminocyclitol-modifying enzymes specified by chromosomal genes in Staphylococcus aureus. AB - A genetic analysis of aminocyclitol resistance in two strains of Staphylococcus aureus was performed. Resistance of strain FK170, isolated in Zurich, was due to the production of a 3'-phosphotransferase [APH(3')]. Strain 5532, isolated in London, produced a 2"-phosphotransferase [APH(2")] and a 6'-N-acetyltransferase [AAC(6')]. Plasmid deoxyribonucleic acid (DNA) was isolated by isopycnic centrifugation from the two parent strains, as well as from susceptible variants and from resistant transductants of both strains. Comparative analysis of plasmid and from resistant transductants of both strains. Comparative analysis of plasmid DNA by centrifugation in sucrose gradients revealed that strain FK170 harbored a 2.7-megadalton tetracycline R-plasmid and a 36-megadalton cryptic plasmid. Strain 5532 contained an 18.5-megadalton penicillinase plasmid. No evidence for plasmid location of the markers for aminocyclitol resistance could be obtained. PMID- 6271047 TI - Effect of subinhibitory concentrations of mecillinam on the serum susceptibility of Escherichia coli strains. AB - For serum-resistant clinical isolates of Escherichia coli were grown in the presence of various subinhibitory concentrations of mecillinam or pivmecillinam and then exposed to the bactericidal action of human serum. All strains became more serum susceptible as a result of pregrowth in medium containing mecillinam, but the concentration of antibiotic needed to produce the effect varied according to the strain being used. Production of ovoid or round cells was a prerequisite for sensitization to serum. Growth in the presence of mecillinam did not alter the response to serum of a serum-susceptible E. coli strain. PMID- 6271048 TI - Activity of moxalactam and cefotaxime alone and in combination with ampicillin or penicillin against group B streptococci. AB - The activities of moxalactam and cefotaxime, alone and combined with ampicillin or penicillin, against 40 isolates of group B streptococci were assessed by using the microtiter broth dilution, checkerboard, and time-kill techniques. Penicillin and cefotaxime were bactericidal for all isolates at concentrations of 0.06 micrograms/ml or less. Ampicillin was slightly less active. Moxalactam was bactericidal for all strains at concentrations of 4 to 8 micrograms/ml. The ampicillin- moxalactam combination was partially synergistic for 60% of the isolates tested; the ampicillin-cefotaxime combination was partially synergistic for 35% of these isolates. No instances of antagonism were observed. In time-kill evaluations, ampicillin (3.0 micrograms/ml) and penicillin (0.75 micrograms/ml) effected 2.5 to 3.5 log10 reductions in numbers of colony-forming units. The addition of 4 micrograms of cefotaxime per ml or 8 to 16 micrograms of moxalactam per ml to penicillin or ampicillin did not alter killing kinetics. Moxalactam and cefotaxime neither enhanced nor decreased the activity of ampicillin or penicillin against group B streptococci. PMID- 6271049 TI - Treatment of gonorrhea: comparison of cefotaxime and penicillin. AB - Ninety-seven patients with 118 sites infected with Neisseria gonorrhoeae were treated with a single dose of either procaine penicillin G (4.8 x 10(6) U) or cefotoxime (1.0 g) intramuscularly. Only the penicillin group took 1 g of probenecid orally. The numbers of infected sites in each treatment group were as follows: penicillin-urethra, 37; rectum, 9; cervix, 8; and pharynx, 4; cefotaxime urethra, 42; rectum, 9; cervix, 5; and pharynx, 4. The cure rates in each treatment group were 100%. No adverse reactions were noted in either group. beta Lactamase-positive N. gonorrhoeae strains were not found. Ninety-five percent of clinical isolates were inhibited by less than or equal to 0.007 micrograms of cefotaxime and less than or equal to 0.25 micrograms of penicillin per ml. In this study cefotaxime was as effective as procaine penicillin in the treatment of uncomplicated gonorrhea. PMID- 6271050 TI - Inhibition of mouse LM cell replication by trifluorothymidine: role of cytosolic deoxythymidine kinase. AB - The effects of trifluorothymidine (5-trifluoromethyl-2'-deoxyuridine, F3dThd) on the replication of three mouse cell lines, LM929, Ltk- (and LM929 derivative devoid of cytosolic deoxythymidine [dThd] kinase activity), and Ltk- c139 (a Ltk- derivative which expresses herpes simplex virus type 1-specified dThd kinase subsequent to biochemical transformation with ultraviolet-irradiated herpes simplex virus type 1), have been investigated. Complete inhibition of Ltk- cell growth required a 10,000-fold higher concentration of F3dThd (1.0 mM) than was required to completely inhibit LM929 and Ltk- c139 cell growth. The plating efficiency of exponentially dividing Ltk- cells after exposure to F3dThd (10 microM) for 24 h was 63% as compared to 3% for exponentially dividing LM929 cells. Stationary LM929 cells (confluent cultures held for a 6-day period in serum-reduced medium) with reduced dThd kinase specific activity and deoxyribonucleic acid biosynthesis level exhibited a plating efficiency similar to that of exponentially dividing Ltk- cells after exposure to F3Thd (1.0 mM) for 24 h. In addition, treatment of exponentially dividing LM929 and Ltk- cells with F3dThd (10 microM) for 24 h resulted in approximately an 80% and 25% reduction in deoxyribonucleic acid biosynthesis, respectively. These data indicated a requirement for cytosolic dThd kinase in the expression of F3dThd-induced cytotoxicity. F3dThd was shown to be a linear competitive inhibitor with respect to dThd for affinity-purified LM929 cytosolic dThd kinase. The Km(app) for dThd and Ki(app) for F3dThd with the cytosolic dThd kinase were 2.4 and 3.8 microM, respectively. PMID- 6271051 TI - In vitro and in vivo morphological response of Klebsiella pneumoniae to cefotiam and cefazolin. AB - The effect of cefotiam and cefazolin on the ultrastructure of Klebsiella pneumoniae DT-S in vitro and in experimental pneumonia in mice was examined by electron microscopy. The action of both cephalosporins against K. pneumoniae DT-S was bactericidal, and a dose response in the action was definite. At the minimal inhibitory concentration of each cephalosporin, filamentation of the cells was induced and the cytoplasm became sparse during the course of incubation. With elevation of the concentration of the cephalosporins, spheroplasts were formed; they subsequently collapsed. In the lungs of mice, the infecting organisms localized in the alveolar space, and each cell was connected by a threadlike material. A fibrous matrix, located on the cell surface of the infecting organisms, was observed in ultrathin sections. By administration of each cephalosporin to mice, several morphological changes, similar to those noted in vitro, were observed in the infecting organisms. PMID- 6271052 TI - In vitro susceptibility of Haemophilus influenzae and neisseria gonorrhoeae to Ro 13-9904 in comparison with other beta-lactam antibiotics. AB - Ro 13-9904 has high in vitro activity, as does cefotaxime, against 57 Haemophilus influenzae and 60 Neisseria gonorrhoeae strains, including 5 and 11 beta lactamase-producing strains in each group, respectively. PMID- 6271053 TI - Selective inhibition of herpesvirus deoxyribonucleic acid synthesis by acycloguanosine, 2'-fluoro-5-iodo-aracytosine, and (E)-5-(2-bromovinyl)-2' deoxyuridine. AB - The selectivity of inhibition of herpesvirus deoxyribonucleic acid synthesis by acycloguanosine, 2'-fluoro-5-iodo-aracytosine, and (E)-5-(2-bromovinyl)-2' deoxyuridine was determined by isopycnic banding of (32)P-labeled deoxyribonucleic acid from herpesvirus-infected and uninfected cells. PMID- 6271054 TI - Effect of acyclovir on latent herpes simplex virus infections in trigeminal ganglia of mice. AB - The inhibition by acyclovir of the in vitro reactivation of herpes simplex virus from latently infected ganglion explant cultures is dependent on the continuous presence of this drug. Administration of acyclovir subcutaneously, orally, or by continuous perfusion to mice with established latent infections did not eliminate latent virus from the trigeminal ganglia. PMID- 6271055 TI - Alterations of immunoreactive beta-endorphin in the third ventricular fluid in response to electrical stimulation of the human periaqueductal gray matter. AB - Immunoreactive beta-endorphin in the third ventricular fluid was measured in response to electrical stimulation of the periaqueductal gray matter in 8 patients with intractable pain during rostral mesencephalic reticulotomy for pain relief. In all patients, marked increase of immunoreactive beta-endorphin was observed. On the other hand, in cases of electrical stimulation of the zona incerta performed during stereoencephalotomy, in 5 patients with involuntary movement, immunoreactive beta-endorphin in the third ventricular fluid did not show any significant change. The authors conclude that the increase of immunoreactive beta-endorphin on electrical stimulation of the periaqueductal gray matter is not a nonspecific response to brain stimulation but a specific response in regard to cerebral localization of endorphins. Direct correlation between pain relief and periaqueductal gray stimulation is also questioned. PMID- 6271056 TI - Transformation of bile acids by Clostridium perfringens. AB - Thirty-five strains of Clostridium perfringens were examined for their ability to transform bile acids, both in growing cultures and by washed whole cells. All of the strains oxidized the 3 alpha-hydroxy group to an oxo group, and all except three converted the same alpha-hydroxy group into a beta-configuration. The oxidative 3 alpha-dehydrogenation was barely detectable under anaerobic cultural conditions but was clearly demonstrated in an aerated system using washed whole cells, with a pH optimum between 7.0 and 9.0. The epimerizing reaction amounting to 10 to 20% conversion was observed in anaerobic cultures and also with resting cells, irrespective of oxygen supply. Both reactions were carried out with seven conventional 3 alpha-hydroxy bile acids, thus producing a series of 3-oxo and 3 beta-hydroxy derivatives that could be examined for gas-liquid chromatographic and mass spectrometric behavior. No evidence for the occurrence of 7 alpha- and 12 alpha-hydroxysteroid dehydrogenase activities among the test strains was found. A highly potent deconjugating hydrolase was elaborated by all of the strains. PMID- 6271057 TI - Detection of viruses in drinking water by concentration on magnetic iron oxide. AB - Discharge of raw domestic wastes containing human enteric viruses into water courses, consumption of untreated water from canals, streams, and shallow wells in villages, and cross-contamination of water in the distribution system because of intermittent water supply in urban areas continue to cause widespread outbreaks of infectious hepatitis in India. To detect a low number of viruses in 50- to 100-liter samples of water, a method was developed with magnetic iron oxide as the virus adsorbent. Poliovirus-seeded dechlorinated tap water, adjusted to pH 3.0 and 0.0005 M AlCl3, was filtered through a 10-g bed of iron oxide sandwiched between two AP20 prefilter pads held in a 142-mm-diameter, stainless steel holder. Virus was eluted from iron oxide by recirculating three times a 100 ml volume of 3% beef extract, pH 9.0. The eluate was reconcentrated to 5 ml by adjusting to pH 3, adding 1 g of iron oxide, stirring for 30 min, and eluting the readsorbed virus with 5 ml of beef extract, pH 9.0. Virus recovery varied from 60 to 80%. Using the above method, we took a survey of drinking water at three locations in Nagpur during 1976 and found the presence of virus in 7 of 50 samples. The quantity of virus recovered ranged from 1 to 7 plaque-forming units per 30 to 60 liters. Virus was detected in some samples even with residual chlorine. No coliforms were detected in the virus-positive samples. PMID- 6271058 TI - Inactivation of poliovirus by chloramine-T. AB - Since concern has recently been expressed about the presence of genotoxic substances due to chlorination of water and wastewater, chloramine-T (CAT) is proposed as an alternative disinfectant to chlorine. The viricidal properties of chlorine and CAT were compared. Kinetics of inactivation of poliovirus type 2 by chlorine and CAT in chlorine demand-free water were investigated by using a kinetic apparatus. Inactivation of the virus by chlorine and CAT occurred in two steps. The initial linear part of the inactivation curve followed a pseudo-first order reaction with the virus. An obvious dose-response relationship was demonstrated with CAT. The rate of inactivation of the virus by CAT was faster in acid medium than in alkaline medium. Inactivation kinetic studies were performed at different temperatures, and the kinetic, Arrhenius, and thermodynamic parameters were evaluated. The rate of inactivation of poliovirus type 2 by chlorine was faster than that by CAT under identical conditions. A mechanism for the viral inactivation in acid conditions was proposed which led to a rate equation consistent with the experimental results. The results indicate that CAT may be an effective viricide against poliovirus type 2 in an acid medium. PMID- 6271059 TI - Growth of Peptococcus and Peptostreptococcus: effect of variations of culture media on efficiency of recovery. AB - Reference strains and clinical isolates of Peptococcus and Peptostreptococcus spp. were evaluated for their growth response in supplemented thioglycolate-yeast extract media. Supplements used included various combinations of hemin, menadione, sodium bicarbonate, and Tween 80. Parallel studies were done to compare the efficiency of recovery of viable cells grown in thioglycolate-based media and Wilkins-Chalgren broth and agar. In addition, the effects of age of the medium and medium storage on viable cell yields for reference strains were determined. Reference strains grown in freshly prepared thioglycolate-yeast extract medium supplemented with sodium bicarbonate produced a 10-fold greater increase in the number of viable cells recovered after 24 h of incubation than did the same organisms cultivated in Wilkins-Chalgren medium. The efficiency of recovery of organisms when either mid-logarithmic- or mid-stationary-phase cells were used to prepare standardized inocula was similar. The results suggest that thioglycolate-yeast extract medium supplemented with sodium bicarbonate is more productive than Wilkins-Chalgren medium for the cultivation of anaerobic gram positive cocci and may represent a suitable alternative for antimicrobial susceptibility testing of these organisms. PMID- 6271060 TI - 1 alpha, 25-difluorovitamin D3: an inert vitamin D analog. PMID- 6271061 TI - Inhibition of rabbit liver fructose 1,6-biphosphatase by AMP: effect of temperature and physiological concentrations of cations and anions. PMID- 6271062 TI - The covalent structure of pig kidney fructose 1,6-bisphosphatase: sequence of the 60-residue NH2-terminal peptide produced by digestion with subtilisin. PMID- 6271064 TI - The effect of tunicamycin on secreted glycosidases of Aspergillus niger. PMID- 6271063 TI - Binding proteins for vitamin D metabolites: serum carriers and intracellular receptors. PMID- 6271065 TI - Mutual regulation between mitochondrial ATPase and respiratory chain activities. PMID- 6271066 TI - The effect of diabetes, nutritional factors, and sex on rat liver and kidney mevalonate kinase, mevalonate-5-phosphate kinase, and mevalonate-5-pyrophosphate decarboxylase. PMID- 6271067 TI - Substrate specificity of Gaucher spleen phosphoprotein phosphatase. PMID- 6271068 TI - Preparation of highly radioactive homogeneous 32P-labeled hydroxymethylglutaryl coenzyme A reductase from rat liver. PMID- 6271069 TI - Biochemical properties of A-homovitamin D derivatives: 1,4-dihydroxy-3-deoxy-A homo-19-nor-9, 10-secocholesta-5,7-dienes. PMID- 6271071 TI - EPR studies of a nonphotosynthetic mutant of Rhodospirillum rubrum. PMID- 6271070 TI - Characterization of bacteriophage T4 and D RNA, a low-molecular-weight RNA of unknown function. PMID- 6271072 TI - Circular dichroism spectra of some lower homologs of bradykinin potentiating peptide 9 alpha. PMID- 6271073 TI - Studies of the kinetics of oxidation of cytochrome c by cytochrome c oxidase: comparison of the reactions of purified and membrane-bound oxidase. PMID- 6271074 TI - Regulation of C4 photosynthesis: mechanism of activation and inactivation of extracted pyruvate, inorganic phosphate dikinase in relation to dark/light regulation. PMID- 6271075 TI - Rabbit liver fructose 1,6-bisphosphatase: labeling of the active and allosteric sites with pyridoxal 5-phosphate and sequence of a nonapeptide from the active site. PMID- 6271077 TI - Pigmented eccrine acrospiroma. Report of a case. PMID- 6271076 TI - Therapeutic considerations in recessive dystrophic epidermolysis bullosa. PMID- 6271078 TI - [Nuclear inclusions in malignant melanoma of the uvea]. PMID- 6271079 TI - Agricultural chemical use and congenital cleft lip and/or palate. AB - A case-control study utilizing vital records and ecologic, surrogate exposure measures was conducted in Iowa and Michigan. The study hypothesis anticipated an excess risk of clefts among fetuses exposed during the peak agricultural chemical use period (April through November) coincident with their first trimester of gestation. To examine this hypothesis, multiple regression techniques were used to aid identification of potential confounders; additional analyses, stratified on the potential confounders, were performed using two chemical exposure indices. The major findings of these analyses suggest: (1) an agricultural chemical effect (using the multiple exposure index) controlling for season of conception; (2) no independent effect of season of conception (thus the null hypothesis is not rejected); and (3) little chemical/season interaction. These results imply that if exposures to agricultural chemicals are, in fact, risk factors for clefts, an expanded model that accounts for multiple pesticidal exposures may be more sensitive than consideration of season of exposure, as originally hypothesized. PMID- 6271080 TI - A wire reclamation incinerator as a source of environmental contamination with tetrachlorodibenzo-p-dioxins and tetrachlorodibenzofurans. AB - The authors investigated an outbreaks of unusual illnesses in humans and horses residing within 1.3 km of a wire reclamation incinerator. The study included site visits; medical and veterinary examinations; analyses of furnace ash, fly ash, soil, and biologic samples for air residues. Tetrachlorodibenzo-p-dioxins (TCDDs) and tetrachlorodibenzofurans (TCDFs) were discovered in furnace ash, fly ash, soil, horse fat, and horse liver samples. PMID- 6271081 TI - Inhibition of simian virus 40 growth in a kidney cell line by an endogenous inhibitor of renal cell proliferation. PMID- 6271082 TI - [Acute respiratory syndromes caused by viruses in a pediatric department. Clinical and epidemiological studies of a series of cases admitted in the period from February to May, 1979]. AB - In the period between February and May 1979, in concomitance with a sharp increase in admittance to the pediatric Division of the Parma General Hospital for acute respiratory infections, we carried out a virologic and serologic investigation on hospitalized children aged 21 days-12 years. The investigation was carried out on 137 patients with respiratory diseases: 33 with bronchiolitis, which occurred mainly (32/33) between February and March in children that were prevalently (30/33) in the first year of life; 27 with upper respiratory tract infections; 54 with mid respiratory tract infections; 23 with pneumonia. Forty eight controls (without respiratory infections) were also investigated. In the months of February and March there was a high circulation of respiratory syncytial virus (RSV): throughout the entire period there was a lower incidence of Adenovirus infections and more uniformly distributed. PMID- 6271083 TI - Prognosis in stage II (T1N1M0) breast cancer. AB - As part of a detailed study of prognostic factors in breast cancer, we have analyzed the ten year survival rates of 524 patients with primary invasive carcinomas 2.0 cm or less in diameter (T1). This report describes the subset of 142 patients (27%) who had metastases only in axillary lymph nodes (T1N1M0). All the patients were treated initially by at least a modified radical mastectomy. Factors associated with a significantly poorer prognosis were: axillary lymph node metastases suspected on clinical examination; perimenopausal menstrual status at diagnosis; tumor larger than 1.0 cm; prominent lymphoid reaction; infiltrating duct or lobular rather than medullary, colloid and tubular carcinoma; and blood vessel invasion. When compared with those patients with negative nodes (T1N0M0), the patients with one or more lymph node metastases had a significantly poorer prognosis. Generally, survival rates tended to diminish as the number of involved lymph nodes increased. In this respect, comparison of patients with one-three and four or more nodal metastases provided a significant discrimination of prognostic groups in the entire series. However, for patients with disease limited to Level I, the same discrimination was obtained comparing those with one-two and three or more positive nodes. In the subset with a single lymph node metastasis, the size of the metastasis (micro or less than or equal to 2 mm vs macro or greater than 2 mm) was not significantly related to prognosis. Lymph node metastases were significantly less frequent among tumors smaller than 1 cm and special tumor types (medullary, colloid, lobular and tubular). However, no factor proved to be a reliable predictor of the presence of axillary metastases for the single largest group consisting of patients with infiltrating duct carcinoma 1-2 cm in diameter. PMID- 6271084 TI - Isolation and partial characterization of active polysomes from calcified and matrix-containing tissues. AB - Polysomes were isolated from calcified and matrix-containing tissues, such as rat calvaria, rat chondrosarcoma and chick embryos. The method of isolation involves preliminary swelling of the tissues in hypotonic buffer containing heparin and cycloheximide. After homogenization, differential centrifugation is used to separate membrane-bound and non-bound polysomes. Each fraction is rehomogenized in the presence of detergent (Triton X-100) and potassium ions (0.25M). Polysomes are harvested by centrifugation through sucrose cushions in the continued presence of high levels of potassium ions and heparin. Total, non-bound, and membrane-bound polysomes prepared in this manner are equally active in protein synthesizing activity in an heterologous cell-free system. The distribution between non-bound and membrane-bound polysomes in the 12 day old chick embryo is 43 and 57 per cent respectively. Sucrose gradient profiles of polypeptide chains on polysomes labeled in organ culture correlate well with the protein synthetic activity of the isolated polysomes. Much of the protein synthetic activity is devoted to collagen. Polysomal fractions obtained from sucrose gradients show preferential incorporation of 3H-proline and nearly 60 per cent of trichloroacetic acid precipitable material is susceptible to collagenase digestion. Products of synthesis are also substrates for collagen specific enzyme, prolyl hydroxylase. The method described in this communication overcomes the inherent difficulties in obtaining active polysomes from calcified and matrix containing tissues. PMID- 6271086 TI - Some histochemical and experimental studies on the adrenal gland of a percoid fish, Trichogaster fasciatus (Bloch & Schneider). PMID- 6271085 TI - Special capabilities of the sexes: differences in the molecular repertoire of mammal brain, heart and skeletal muscle. AB - World records for men's and women's (anaerobic) 200 meter dash are 19.67 and 21.9 seconds respectively. Here the transphosphorylation mechanisms are operative and not the glycolytic anaerobic and aerobic ones. The purpose of this paper is to try to find molecular differences during the neuro-muscular sexual differentiation in the Wistar rat. Creatine kinase (CPK: EC 2.7.3.2) as model transphosphorylase and the protein synthesis pattern, determined from the post 125,000 x g cytosol fraction of the brain, heart and masseter muscle, were chosen as molecular monitors. The CPK specific activity profiles showed differences among tissues and sexes. In rats with similar weights between 200 and 320 g, the epigenetic control of protein synthesis in the studied tissues, reflected genome differential activation depending on sex and age. The electrophoretograms differed in regard to the number and position of the protein bands of homologous tissues of different sexes. As an original finding, against the generally accepted criterion (the brain fundamentally synthesizes BB-CPK isoenzyme), MB-CPK and MM-CPK isoenzymes appeared in the brain of both sexes. The molecular sexual differences in the studied tissues determine functional and anatomical differences. PMID- 6271087 TI - [Determination of normal pressure in 15756 African school children from 11 to 18 years old in the Ivory Coast. Comparative study of an African group and an European group]. AB - Blood pressure readings of 15756 coloured African schoolchildren aged between 11 and 18 years old were studied. The measurements were casual readings. The study group was not perfectly representative of the school population. However, from the practical point of view, the data may be used in the whole Ivory Coast school population in the 11 to 18 years age range. The distribution of the average readings with respect to age and sex is indicated on Table II. Abnormally high blood pressure was defined as readings greater than the average BP + 2 standard deviations, to the nearest 0,5 cm Hg (to take the practical problems of BP measurement into account). Under these conditions, all boys and girls between 11 and 18 years of age with BP greater than or equal to 150/90 (except boys of 17-18 years of age for whom the limit was 155/95) were considered to be hypertensive. According to these criteria the prevalence of hypertension in coloured African schoolchildren was: -systolic hypertension: 4,86% boys and 4,03% girls; diastolic hypertension: 4,20% boys and 5,09% girls; -systolic or diastolic hypertension: 6,77% boys and 7,54% girls; -systolic and diastolic hypertension: 2,29% boys and 1,57% girls. In fact, these would be children "at risk" rather than hypertensive. The true number of hypertensive children would only become apparent on follow-up. A comparison with coloured African and white European schoolchildren in the same schools in Abidjan showed that the average BP readings and prevalence of hypertension was higher in the coloured population but the difference was not always statistically significant. The small number of European children made comparison difficult. The data obtained was compared to previously reported series but it was difficult to draw conclusions because of differences in methodology. PMID- 6271088 TI - [Importance of radioreceptor assays in determining the mechanism of action of hypotensive alpha-adrenergic compounds]. AB - In rat central nervous system, 3H-prazosin and 3H-clonidine bind to two different classes of alpha-adrenoceptors: - 3H-prazosin binds to alpha1-adrenoceptors with high affinity (KD 25 degrees C = 0.30 nM) with a number of binding sites of 81.6 fmoles/mg of protein. -3H-clonidine binds to alpha 2-adrenoceptors with high affinity (KD 25 degrees C - 2.1 nM) with a number of binding sites of 190.0 fmoles/mg of protein. The radioreceptor assay may help to determine rapidly in vitro the neurochemical profile of new antihypertensive compounds. Thus S3341 will exert its hypotensive action at the level of alpha 2-adrenoceptors as clonidine (Catapressan*) but with a specificity 2-3 times greater. PMID- 6271089 TI - Receptor sensitivity and the mechanism of action of antidepressant treatment. Implications for the etiology and therapy of depression. AB - Considerable evidence suggests that the acute effects of antidepressant treatments on brain norepinephrine (NE) and serotonin (5-HT) systems cannot account fully for their delayed therapeutic action. This review evaluates the effects of long-term antidepressant treatment on biogenic amine metabolism and on various indexes of presynaptic and postsynaptic receptor function. In contrast to variable effects on NE and 5-HT turnover and on presynaptic receptor sensitivity almost all long-term antidepressant treatments produce consistent alterations in a number of measures of postsynaptic amine receptor sensitivity. Long-term treatment has been found to reduce beta-adrenergic sensitivity while enhancing responses to serotonergic and alpha-adrenergic stimulation, suggesting that modulation of receptor sensitivity may be a mechanism of action common to tricyclic antidepressants, "atypical" antidepressants, monoamine oxidase inhibitors, and electroconvulsive therapy. These findings provide support for hypotheses of amine receptor abnormalities in depression and indicate the need for expanded studies of amine receptor function in patients. PMID- 6271090 TI - Leukocyte migration inhibitory factor (LMIF) release by human colonic lymphocytes. AB - Lamina proprial lymphocytes (LPL), isolated by an EDTA-collagenase technique from patients with various colonic diseases, were investigated for LMIF release in vitro. On stimulation with the preparation of Kunin antigen, macrophage-depleted LPL from patients with severely or moderately active ulcerative colitis showed LMIF release which was significantly greater than that observed using LPL from patients with mild colitis or from those with other diseases of the large bowel, including Crohn's disease. Results similar to those obtained with LPL were found with the corresponding peripheral blood lymphocytes (PBL) stimulated by the preparation of Kunin antigen. In contrast, nonspecific stimulation in vitro with Concanavalin A showed no differences in LMIF releases by the LPL or PBL in the various disease groups. It is suggested that hypersensitivity to Kunin antigen may have pathogenic significance in ulcerative colitis. PMID- 6271091 TI - Immunohistochemical identification of factor VIII-related antigen in the intravascular bronchioloalveolar tumor of the lung. PMID- 6271092 TI - [Synthesis of 3,3'disubstituted hexestrol derivatives and their binding affinity to cytoplasmatic estrogen receptors (author's transl)]. PMID- 6271093 TI - [Effect of various enzyme-substrate ratios on trypsin activity in the presence of trace elements]. AB - In vitro experiments were conducted with trypsin and Na-Benzoyl-l-arginine-p nitroanilid (L-BAPA) as substrate. The aim of the work was to explore the influence of additional Cu2+, Zn2+, Co2+, Ni2+ and Fe3+ ions in the reaction mixtures on the trypsin activity at different enzyme-substrate ratios (E : S = 1 : 100; 1 : 50; 1 : 10; 1 : 5). The reaction conditions were chosen according to the steady-state conditions of the tryptic hydrolysis. The trypsin activity increased by adding small amounts (4.5 X 10(-7) mol Cu2+/l) and decreased by adding higher amounts of Cu2+ ions. There was no influence of the E : S ratio. The tryptic activity was only partly influenced by the substrate concentration when the other trace elements were added to the reaction mixtures. These results may explain in part the improved protein digestibility which is reported when 250 mg Cu/kg are added to pig feeds. PMID- 6271094 TI - Nephrotoxicity of cefotiam (CGP 14221/E) in rats and rabbits. AB - Cefotiam (CGP 14221/E; SCE 963), a semisynthetic cephalosporin, was administered as a single dose by i.v. injection to rats l(up to 1.8 g/kg body-weight) and rabbits (up to 1.7 g/kg body-weight). Cephaloridine served as positive control (1.0 and 0.75 g/kg in rats; 0.3 and 0.28 g/kg in rabbits). The animals were sacrificed 24 h after injection and the kidneys preserved for routine histology and enzyme histochemistry (alkaline phosphatase, aminopeptidase, succinate dehydrogenase, esterase). Serum samples (Na+, K+, Cl-, urea, creatinine, LDH, alkaline phosphatase) and 24-h urine (Na+, K+, Cl-, urea, creatinine, protein, LDH, aminopeptidase) were analysed before and 24 h after injection. Minimal, irregularly scattered, degenerative changes in the proximal tubules which were not dose-dependent in degree were observed in rat kidneys following cefotiam injection. A slight dose-dependent degeneration in up to 50% of proximal tubular cells with loss of brush-border membrane enzyme activity was observed in rabbit kidneys. In both animal species the ability to concentrate urine was retained and urea and creatinine serum levels hardly affected. Following cefotiam injection a dose-dependent 4-fold excretion of urinary protein but not of LDH was observed in rabbits only. By contrast, cephaloridine caused extensive degeneration and necrosis in up to 90% of proximal tubular cells in both rats and rabbits, which was accompanied with formation of enzymically active hyaline casts, loss of urine concentrating capacity of the kidney, elevated serum levels of urea and creatinine and an increased urinary excretion of LDH (60-fold in rats, 20-fold in rabbits) and protein (3-fold in rats, 10-fold in rabbits). Histochemistry and electron microscopy of rabbit kidneys suggested a loss of microvilli from proximal tubule cells by endocytosis and thus degeneration following injection of large doses of cefotiam, whereas cell disruption and necrosis prevailed after cephaloridine. The action of cefotiam on the proximal tubule cells is, therefore, not only quantitatively but possibly also qualitatively different from that of cephaloridine. Semiquantitative evaluation of tubular injuries in alkaline phosphatase-stained kidney sections and measurements of LDH and protein content in 24-h urine samples were advantageous in determining the quantity and the quality of nephrotoxic effects. PMID- 6271095 TI - The genetics of aphthovirus. Brief review. PMID- 6271096 TI - Antigenicity of mouse hepatitis virus strain 3 subcomponents in C57 strain mice. AB - C57 strain mice were inoculated intraperitoneally with denatured mouse hepatitis virus strain 3 particles and virus surface projection, membrane and ribonucleoprotein subcomponents, obtained from detergent treated purified virus preparations. All immunised animals developed high levels of serum antibody directed against the respective antigens, detectable by enzyme-linked immunosorbent assay. Mice that had been immunised with denatured virus particles or surface projections were protected against infection with mouse hepatitis virus strain 3, whereas immunisation with virus membrane or ribonucleoprotein subcomponents failed to protect mice against virus challenge. PMID- 6271097 TI - Persistent reactivable latent herpes simplex virus infection in trigeminal ganglia of mice treated with antiviral drugs. AB - Latent reactivable infection was established with HSV in mouse trigeminal ganglion. A number of antiviral drugs (IUDR, acycloguanosine, Ara-A, PAA and PFA), effective against acute CNS infection with HSV, failed to influence the latent infection once established. Neither lipophilic properties of PFA derivatives nor the combination of Ara-A and acycloguanosine improved the drug effects on HSV latency. PMID- 6271098 TI - Isolation of cytopathic porcine rotavirus in cell roller culture in the presence of trypsin. AB - Cytopathic porcine rotavirus was isolated in roller tube cultures of MA-104 cells. Faeces of a piglet suffering from diarrhoea, were treated with trypsin which was also added to the maintenance medium. Using stationary cultures, virus was not isolated from the same materials. The cytopathic effect was clearly observed after 8 serial passages and the virus titer at the 14th passage level was 10(7) median tissue culture infective doses per ml, or higher. At the 27th passage, rotavirus particles were seen by negative contrast electron microscopy at a density of 1.36 to 1.38 g/cm3 in CsCl centrifugation gradients. There are partial cross-neutralization between the Lincoln strain of bovine rotavirus and porcine rotavirus from pigs or tissue cultures. Gnotobiotic piglets were inoculated with pig-passaged virus and viral antigen was detected in epithelial cells of small intestinal villi by immunofluorescence. The highest titer of virus was detected in faeces 72 hours after inoculation. The cell culture adapted virus which produce a cytopathic effect was designated the strain S 80 of porcine rotavirus. PMID- 6271099 TI - Stabilization of isometric DNA viruses against thermoinactivation by lowered ionic strength. AB - The thermostability of isometric DNA viruses increases, if the ionic strength is diminished before heating. When unpurified virus material from cell cultures is heated under conditions (temperature, time) which lead to a reduction in infectivity by 3 to 4 log10, this loss in titer is 1 to 3 log10 less, if the ionic strength is decreased by diluting the material in dist. water (1:100) before heating. A dilution in Eagle's MEM (1:100) or a previous dialysis against water does not have the same effect. This property was found in the following members of 4 DNA virus families: adenovirus 5, herpesvirus 1, SV 40 and bovine parvovirus 1. In contrast, members of pox- (vaccinia) and picornaviruses (polio 2, coxsackie B5) were found to be less thermostable under conditions of low ionic strength. Reovirus 3 showed no difference in inactivation. The observed effect may be of practical importance for heat disinfection of viruses and for their persistence in the environment. PMID- 6271100 TI - Structural characterization of the major internal protein of the SKA 21-3 virus. Brief report. PMID- 6271101 TI - Biological and biochemical investigations on five European isolates of Mamestra brassica nuclear polyhedrosis virus. AB - Five multiply enveloped European isolates of Mamestra brassicae nuclear polyhedrosis virus (Oxford, German, French, Dutch and Danish) were found to be very closely related serologically using the enzyme linked immunosorbent assay (ELISA) double antibody sandwich method and immunodiffusion. By SDS polyacrylamide gel electrophoresis of viral proteins and restriction endonuclease analysis of DNA using seven enzymes there appeared to be two variants as the Oxford and German isolates were distinct from the other three. The German isolate was shown to be more susceptible to Nonidet P40 detergent treatment affecting some nucleocapsid structural polypeptides which also reduced antigenicity in gel immunodiffusion plates. In bioassays of polyhedra, the Dutch isolate showed a higher LD50 than the other viruses although this was not statistically significant. PMID- 6271102 TI - Replication of porcine enteric adenoviruses in vivo. AB - The replication of an enteropathogenic porcine adenovirus in the intestinal epithelial cells of naturally and experimentally piglets has been studied by transmission electron microscopy (TEM) and by immunoperoxidase (Ip) staining of paraffin sections. Three types of intranuclear inclusion bodies were observed. Viral particles appeared to be assembled from electron dense crescents which seemed to originate from type II intranuclear inclusion bodies. Virus accumulated in the nucleus of infected cells. It formed paracrystalline arrays in the electron dense centre of the nucleus but was dispersed in the peripheral electron translucent zone. Virus was released from the cells after rupture of the nuclear and plasma membranes. The results are compared with published data on the replication of adenoviruses in cell cultures. PMID- 6271103 TI - Replication of poliovirus and measles virus in cultures of human lymphoblastoid and of Burkitt lymphoma cell lines. AB - Poliovirus type 1 replicated in 4 different human lymphoblastoid cell lines (LBL) transformed in vitro by EBV virus or isolated from cases of mononucleosis. Maximum virus titers were reached 2--4 days after inoculation. There was a decrease in percentage of viable cells in the infected cultures but a considerable fraction of cells was not destroyed by virus replication. A persistent low grade replication of virus was observed and demonstrable during 56 days after inoculation in one LBL cell line. Presumably a small fraction of cells supporting virus replication is continuously recruited from refractory cells. No virus propagation was demonstrable in 4 Burkitt lymphoma (BL) cell lines. Measles virus grew efficiently in both types of cell lines. Using indirect immunofluorescent technique poliovirus and measles virus antigen could be demonstrated in the cytoplasm of LBL cells in parallel with disappearance of F actin containing microvilli. PMID- 6271104 TI - Oncogenic activity of the BK type of human papova virus in newborn Wistar rats. AB - The oncogenic potential of the human papova virus BK (= BKV) has been examined in newborn Wistar rats. 12 of 37 animals inoculated with BKV s.c. and 7 of 40 animals inoculated with BKV i.c. developed tumors of various histological types. The latency periods ranged from 6 to 18 months. The BKV etiology of tumors was supported by detection of BKV T antigen in cells of established tumor lines by means of indirect immunofluorescence. No tumors appeared in animals given injections of saline instead of virus. BKV T antibodies were detected in sera of 100 per cent of animals bearing tumors but no in sera of control animals. Only 3 of 77 BKV infected and none of the control animals developed spontaneous tumors (adenomas). PMID- 6271105 TI - Genetic resistance to lethal flaviviral encephalitis. III. Replication of Banzi virus in vitro and in vivo in tissues of congeneic susceptible and resistant mice. AB - The replication of Banzi virus, flavivirus, was compared in vitro and in vivo in tissues of congenic mice genetically resistant (C3H/RV) or genetically susceptible (C3H/He) to lethal infection. Ultrastructural changes in brains of resistant and susceptible adult mice following intracerebral or intraperitoneal inoculation of virus also were compared. Banzi virus replicated equally well in monolayer cultures of infant and adult brain, stimulated and non-stimulated macrophages and embryonic cells from both strains of mice. Similarly, no significant differences were found between strains in virus growth in brain, spleen or thymus of peripherally-inoculated infant mice. In intracerebrally inoculated adult mice, virus titers in brains of resistant mice were consistently lower than in susceptible mice. Visualization of virus particles was dependent on virus concentration in tissues. The changes in brain tissues of both strains of mice were similar, differing only in the time of onset which was noted two days later in C3H/RV mice than in C3H/He mice. These results indicate that, in the case of Banzi virus, the phenotypic expression of genetically-determined resistance of lethal flavivirus infection cannot be attributed primarily to the ability of host cells to support virus replication. PMID- 6271107 TI - Food borne disease. PMID- 6271106 TI - Nucleoside triphosphate phosphohydrolase activities in African swine fever virus. AB - African swine fever virus contains nucleoside triphosphate phosphohydrolase activity which releases 32P phosphate from gamma-32P ATP at a rate of about 1 mumol/h mg of virus protein. The hydrolase activity is slightly stimulated by adding nucleic acids to the reaction mixture and under conditions of RNA synthesis. A study of the rate of ATP hydrolysis at different concentrations of ATP suggests the existence of two phosphohydrolase activities with apparent Km values of about 0.04 and 1 mM. PMID- 6271108 TI - Subclinical neuropathy associated with chronic obstructive pulmonary disease: possible pathophysiologic role of smoking. AB - Twenty of 23 patients with chronic obstructive pulmonary disease (COPD) showed electrophysiologic evidence of peripheral nerve dysfunction. Abnormalities of sensory nerve conduction were most common, affecting the sural nerve (20 subjects), ulnar nerve (11), radial nerve (eight), and median nerve (seven). Six subjects had impairment of both sensory and motor nerve function, with the common peroneal being the most frequently affected motor nerve. Clinical signs of neuropathy were found in four patients. Cigarette smoking, expressed as pack years, was correlated significantly with the electrophysiologic abnormalities. These findings indicate that subclinical polyneuropathy commonly occurs in association with COPD and that this COPD-related neuropathy is correlated with cigarette consumption. From these data we suggest that a substance or substances in cigarette smoke, such as nicotine, taken on a long-term basis, may be toxic to peripheral nerves. PMID- 6271109 TI - Surgical experience with nasopharyngeal angiofibroma. AB - Recent reports have suggested that radiotherapy may be preferable to other forms of treatment of juvenile nasopharyngeal angiofibroma. There are, however, potentially serious short- and long-term complications associated with the use of radiotherapy in the head and neck. For the past three years, ten consecutive patients with juvenile nasopharyngeal angiofibroma have been treated at The Cleveland Clinic Foundation with an approach that permits accurate removal with minimal complications. With this method of treatment, intraoperative blood loss, the necessity for blood transfusion, and length of hospital stay have been greatly decreased. There have been no substantive complications and no recurrences to date. If further experience with this approach to management in a larger series of patients has the same results, it would seem that radiotherapy should be relegated to a secondary position in the treatment of juvenile nasopharyngeal angiofibroma, except for cases wherein intracranial extension would prevent total excision. PMID- 6271111 TI - The effect of chronic acid/base disturbances on renal amino acid clearances in the rat. AB - Since the few data available concerning the effect of acid/base disturbances on renal amino acid reabsorption were conflicting, and there were sound theoretical reasons for an effect, we have studied the clearance of endogenous amino acids in the rat in vivo under control conditions and after induction of either metabolic acidosis or alkalosis by administration of NH4Cl or NaHCO3, respectively. The effectiveness of treatment was assessed by examination of plasma and urinary levels of HCO3, Cl, Na and K. It was found that the renal clearance of amino acids, measured during acidosis or alkalosis, did not differ from those found under control conditions, the majority of values being less than 1% of the glomerular filtration rate. Thus, the amino acid reabsorptive mechanism appears unaffected by changes in the pH of the glomerular filtrate and/or by changes in tubular hydrogen ion secretion which would accompany such disturbances. These data are thus in agreement with findings during acidosis in man and in both acidosis and alkalosis in the dog. The findings are contrary to earlier reports from in vitro studies in the rat, and suggest the presence of severe functional impairment in the isolated perfused kidneys used in these earlier studies where very large changes in amino acid clearance were obtained. PMID- 6271110 TI - Possible involvement of parotid beta-adrenergic receptors in the etiology of sialadenosis. AB - The concentration of beta-adrenergic receptors was determined in rat and human parotid glands, in normal tissue as well as after sympathetic denervation of the rat, and in human sialadenosis. Receptor levels were clearly elevated after denervation of the rat and in sialadenosis. The possible implications of these findings for the etiology of human sialadenosis are discussed. PMID- 6271112 TI - Comparative studies on dihydroorotate dehydrogenase from P. berghei and the mouse reticulocyte. AB - Kinetic parameters on dihydroorotate dehydrogenase (DHO-DHase) from the rodent malarial parasite, Plasmodium berghei, have been determined. This enzyme, the fourth in de novo pyrimidine biosynthesis, is particulate and is absent in the mature mammalian red cell. The Km of the substrate, dihydroorotate, was determined to be 23 microM and the Ki values for a number of substrate analogues have been determined. The most potent inhibitor was dihydroazaorotate (Ki, 3 microM), 5-azaorotate (Ki, 20 microM) and other pyrimidine analogues. The activity of the enzyme was also affected by a number of respiratory chain inhibitors. As the P. berghei infection is accompanied by reticulocytosis, a comparative study of DHO-DHase in mouse reticulocytes was also carried out. The general properties of the enzyme from these sources were similar to those of the parasite enzyme. However, significant differences in the response of the two enzymes to various inhibitors were observed and could provide a rational basis for the development of chemotherapeutic agents active against the parasite. PMID- 6271113 TI - Isolation of haemagglutinating adeno-like virus related to virus 127 from an Australian poultry flock with an egg drop syndrome. AB - An egg drop syndrome within Australian broiler poultry is described. The syndrome was characterised by delayed onset of laying, a lower peak in egg production and a drop in egg production shortly after reaching peak production. Antibody to virus 127 was detected in 102 of 106 fowl serums tested. Two haemagglutinating viruses were isolated from one affected flock and one was subjected to further study. It was adenovirus-like on electron-microscopic examination and haemagglutination was not inhibited by a specific antiserum to Newcastle disease virus. An antiserum was raised in White Leghorn fowl against the isolate and this antiserum was found to cross-react with virus 127, a prototype virus of Egg Drop Syndrome 76. PMID- 6271114 TI - Isolation of bluetongue virus serotype 20 from the semen of an experimentally infected bull. PMID- 6271115 TI - A serological survey for antibodies to bluetongue virus in Papua New Guinea. PMID- 6271116 TI - Experimental transmission of avian reovirus and avian adenovirus through embryonated eggs. PMID- 6271117 TI - Kidney, liver, erythrocyte membrane Na, K-adenosine triphosphatase in protein energy malnourished rats. PMID- 6271118 TI - The determination of glycerol concentration and specific radioactivity in rat blood. PMID- 6271119 TI - The relationship between sodium transport and Na+, K+ ATPase in human erythrocytes. PMID- 6271120 TI - Determination of adenosylhomocysteine in urine of immunodeficient children. PMID- 6271121 TI - Inorganic pyrophosphate: D-fructose-6-phosphate 1-phosphotransferase in mung beans and its activation by D-fructose 1,6-bisphosphate and D-glucose 1, 6 bisphosphate. PMID- 6271122 TI - Periodate oxidation studies in the elucidation of the structures of sialic acid containing oligosaccharides. PMID- 6271123 TI - Loss of endogenous nuclear protein A24 lyase activity during chicken erythropoiesis. PMID- 6271124 TI - Transferrin and iron uptake by human lymphoblastoid and K-562 cells. PMID- 6271125 TI - Effects of adrenalectomy and hydrocortisone on DNA synthesis in the rat anterior pituitary gland. PMID- 6271126 TI - Carboxyl terminal tyrosine metabolism of alpha tubulin and changes in cell shape: Chinese hamster ovary cells. PMID- 6271127 TI - Immature red cells have ferritin receptors. PMID- 6271128 TI - Regulation of fructose 2,6-P2 concentration in isolated hepatocytes. PMID- 6271129 TI - Enhancement of ornithine decarboxylase and Na+, K+ ATPase in osteoblastoma cells by intermittent compression. PMID- 6271130 TI - Evidence for a reactive sulfhydryl in the DNA binding domain of the 1,25 dihydroxyvitamin D3 receptor. PMID- 6271131 TI - Effect of 17 beta-estradiol and testosterone on guanosine 3', 5'-cyclic monophosphate in the rat adrenal cortex. PMID- 6271133 TI - Effect of vanadate on the renal accumulation of p-aminohippurate in the rabbit kidney tubules in vitro. PMID- 6271132 TI - Prostacyclin receptors of a neuronal hybrid cell line. Divalent citations and ligand-receptor coupling. PMID- 6271134 TI - Inhibition of gamma-aminobutyric acid stimulated [3H]diazepam binding by benzodiazepine receptors ligands. PMID- 6271135 TI - Inhibition of rat heart superoxidase dismutase activity by diethyldithiocarbamate and its effect on mitochondrial function. PMID- 6271136 TI - Alterations of regional gamma-aminobutyric acid receptors in morphine tolerant mice. PMID- 6271137 TI - Plasma cyclic AMP in the morphine-tolerant rat. PMID- 6271138 TI - Effects of ethanol and lead ingestion on urinary sodium excretion and related enzyme activity in rat kidney. PMID- 6271139 TI - Possible nuclear protein kinase regulation of homologous ribonucleic acid polymerases from small dense nuclei of mouse brain during morphine tolerance dependence. Involvement of cyclic 3',5'-adenosine monophosphate. PMID- 6271140 TI - Biological activities and modes of action of 9-alpha-D-arabinofuranosyladenine and 9-alpha-D-arabinofuranosyl-8-azaadenine. PMID- 6271141 TI - Depression of hepatic cytochrome P-450-dependent mixed function oxidases during infection with encephalomyocarditis virus. PMID- 6271142 TI - Resistance of a separated form of canine ureteral phosphodiesterase activity to inhibition by xanthines and papaverine. PMID- 6271143 TI - Studies in the metabolism of carcinogenic polycyclic heteroaromatic compounds. I. The hepatic microsomal metabolism of 7-methylbenz[c]acridine. PMID- 6271144 TI - Mechanisms of stimulation of rat cardiac muscle by 5-hydroxytryptamine. PMID- 6271145 TI - The effect of organophosphate poisoning on plasma cyclic AMP in rats. PMID- 6271146 TI - [In vitro studies on antacids/Third communication (author's transl)]. AB - With an automated method, four antacid gel formulations were tested for antacid in vitro activity. In addition, some elements involved in the buffer effect were assayed in the solution parallel to the in vitro test. Of the four formulations, one showed only insufficient buffer effect. Compared with the common proton/metal ion exchange of antacid buffering, experimental data of another formulation suggest a partially different proton binding mechanism. PMID- 6271147 TI - [Persistence of transplacental antibodies against rotavirus in children less than 6 months of age]. AB - The persistence of antibodies against rotavirus was studied in the sera of 54 recently born infants up to six months of age; likewise, in the sera of their mothers. Serum positivities were found to be similar in both, showing 96% for those with antibodies and 94.4% for the latter. The percentage of infants with antibodies dropped gradually to the 4th month of age and since then rises in the titre of antibodies began to appear in four infants, which indicated there had been rotavirus infections. These studies lead to believe that in spite of the presence of serum antibodies, rotavirus multiply in the enteric tract and do not bring about serum antibodies, but perhaps, they stimulate the production of coproantibodies, which do not allow in the future the implantation of these viruses in the intestine. PMID- 6271148 TI - [Proline hydroxylase activity and collagen levels in the lungs of chick embryo during ontogeny]. AB - The tropocollagen in the lungs of chicken embryos increases rapidly between the 13th and 16th day of development, then keeps constant levels and equal ones to those of the adult until the birth. The proline hydroxylase activity is highest in the lung of the embryos of 13 days old, diminishes rapidly between the 13th and 16th day, after that moderately but constantly until the birth, when it is equal to the levels found in the adult. The procollagen has a behaviour comparable to the one described for the proline hydroxylase. PMID- 6271149 TI - [Adrenergic control of pancreatic glucagon secretion]. AB - In order to characterize the adrenergic control of pancreatic A cell, the effect on the glucagon secretion of three sympathomimetic substances (epinephrine, isoproterenol, phenylephrine) and two adrenergic blockers (propranolol and phentolamine) have been separately examined by the isolated perfused rat pancreas. The study was performed in basal state and during glucagon hypersecretion induced by arginine or glucopenia. Epinephrine and isoproterenol infusion determined a prompt an sustained glucagon release both in the basal state and during glucagon hypersecretion. The effect of phenylephrine infusion was slight. In the presence of propranolol, glucagon secretion induced by metabolic stimulus was significantly depressed. The glucagon secretion in the same experimental conditions was insignificantly enhanced by phentolamine. Finally propranolol infusion reverse the glucagon secretion induced by phenylephrine. In conclusion the pancreatic glucagon secretion in our model of study is clearly induced by B adrenergic receptor stimulation. PMID- 6271150 TI - [NAD glucohydrolase from bovine retina]. AB - The NAD glycohydrolase activity in the retina is very low compared with the one found in the brain. Therefore the retina extracts ahve a sufficiently high NAD level so that they are able to form "in vitro" lactate from glucose in anaerobiosis in presence of only ATP, Mg++ and glucose. The NAD glycohydrolase has been found in the retina in a great extent in the microsomes. PMID- 6271151 TI - [Effect of heroin administration on MHV-3 virus experimental hepatitis in mice]. AB - Administration of heroin (5mg/Kg/day) in mice for a period of time sufficient to induce dependence and continued during the experiment did not increase susceptibility to MHV-3 virus infection, did not cause more serious forms of hepatitis and did not increase mortality with respect to the controls. PMID- 6271152 TI - [Effect of pentazocine administration on MHV3 virus experimental hepatitis in mice]. AB - Mice treated with 15mg/kg/day pentazocin and infected with MHV-3 virus after 7 days did not show increased susceptibility to MHV-3 virus infection, did not develop more serious forms of hepatitis and mortality did not increase with respect to the controls. Drug administration was continued for the duration of the experiment. PMID- 6271153 TI - [The production of enzymes inactivating various antibiotics obtained from Gram negative bacteria recently isolated in the province of Chieti]. AB - The Authors have studied the ability of producing enzymes blocking various antibiotics by 274 Gram-negative germs isolated in the years 1978-80 in the Province of Chieti (Italy). The results have showed that a high percentage of examined strains presents a polyresistance towards penicillins, cephalosporins, chloramphenicol and amino-glucosides producing in the same time beta-lactamase, acetyl-transferases and phospho-transferases. PMID- 6271154 TI - [Findings of the importance of dietetic fiber in the regulation of cholesterolemia]. AB - We studied the variations of some lipid indexes in cholesterolemia, lipidemia and triglycerides in relation to a diet rich in bulk. Seven adult male subjects in good health were tested for cholesterol, lipid and triglycerides before and after a two week period of a dialy diet of 40 g of whole wheat bran containing about 10g of fiber. At the some time the lipid, cholesterol and triglycerides limits were measured in 3 groups of rabbits. The first groups was subjected to a normal diet, the second group was subjected to a diet of bleached flour and vitamins, and the third group had a diet similar to the second group but with the addition of 40 g whole wheat bran. The results of our investigation demonstrated a statistically significant decrease of cholesterol lipid and triglycerides levels the special diets at the some time we did not see in increase in the third group of rabbit. Our study seems to indicate that dietetic fibers exercise a vital role in regulating the concentration of plasma lipids even if the subject continues to consume a diet rich in fats. From this we see to importance of the dietetic fibers as a protective factor in the prevention of ateriosclerotic. PMID- 6271155 TI - Effect of beta-adrenergic receptor blockade with propranolol on the response of plasma catecholamines and renin activity to upright tilting in normal subjects. AB - 1 Relationship between plasma catecholamines (measured as noradrenaline and adrenaline) and plasma renin activity (PRA) were examined at rest and during passive head-up tilting for 30 min in nine normal subjects, before and after treatment with propranolol 160 mg daily for 7 days. 2 Noradrenaline (NA) and adrenaline (A) increased substantially after tilting for 15 min. There were no changes in PRA. After 30 min tilting, NA remained elevated, whereas A had returned to resting levels. A significant increase in PRA was apparent at 30 min. Pulse rate and diastolic blood pressure increased progressively during tilting. Systolic pressure did not change. 3 Treatment with propranolol reduced pulse rate and systolic blood pressure at rest and during tilting. Resting catecholamine concentrations and the response of NA to tilting were unaffected. In contrast, treatment prolonged the A response leading to significantly higher levels after 30 min tilting. Propranolol reduced PRA in six of the nine subjects and prevented the increase with tilting observed before treatment. PMID- 6271156 TI - Development of tolerance to oral salbutamol in the third trimester of pregnancy: a study of circulatory and metabolic effects. AB - 1 The purpose of the study was to investigate whether a development of tolerance does occur or not to salbutamol-induced cardiovascular and metabolic effects after oral long term treatment in pregnancy. 2 Twenty-three women in late pregnancy were given an oral dose of 4 mg salbutamol in the morning after an overnight fast. Ten of the women had not been exposed to beta-adrenoceptor agonists earlier in their pregnancy (group A). Thirteen of the women had been treated with salbutamol in a dose of 4 mg four times a day for 12 to 33 days preceding the study (group B). 3 Blood samples were collected every 30 min for 120 min and analyzed for cyclic AMP, insulin, C-peptide, glucose, lactate, glycerol, non-esterified fatty acids (NEFA) and 3-hydroxybutyrate (3-HB). Heart rate and blood pressure were recorded simultaneously. 4 In group A there was significant increase in plasma cyclic AMP and increased glycogenolysis, lipolysis and insulin secretion. In this group there were also significant cardiovascular effects, that is, an increase of heart rate and decrease of diastolic blood pressure. 5 In group B the effects of salbutamol were significantly less pronounced, indicating a development of tolerance. 6 We conclude that the metabolic side-effects are not sufficient reason to withhold beta-adrenoceptor agonists to healthy pregnant women with threatened premature labour, since tolerance rapidly develops to these actions. PMID- 6271158 TI - Malignant tumours of liver and lung with midline granuloma in an industrial worker. PMID- 6271157 TI - Lymphocyte cyclic AMP production in the elderly: the effects of prostaglandin E1. PMID- 6271159 TI - Minor histocompatibility antigens do not enhance BALB/c anti-SV40 TASA response. PMID- 6271160 TI - Hydrogen peroxide and superoxide release by alveolar macrophages from normal and BCG-vaccinated guinea-pigs after intravenous challenge with Mycobacterium tuberculosis. AB - The release of H2O2 and .O2- by alveolar macrophages from normal and BCG vaccinated guinea-pigs was measured 3 h, 3 days and 6 days after i.v. challenge infection with Mycobacterium tuberculosis H37Ra. Vaccination did not affect the release of H2O2 or .O2- form macrophages that were removed from guinea-pigs 3 h after i.v. infection and tested as monolayers without a phagocytic stimulus. However, macrophages that were removed from vaccinated animals on the third and sixth days after i.v. infection released progressively more than macrophages that were removed after 3 h. This was not seen with cells from i.v.-infected normal animals. Exposure of macrophage monolayers to phorbol myristate acetate (PMA) and opsonized H37Ra caused increased release of H2O2 and .O2-. There was no difference in the response to PMA either between macrophages from normal animals and those from vaccinated animals or between macrophages taken 3 h, 3 days and 6 days after i.v. infection. Thus the response with PMA gave no indication of the development of local immunity. In contrast, with H37Ra as a phagocytic stimulus in vitro the amounts of H2O2 and .O2- released per cell-associated bacillus increased with the time elapsed since i.v. infection. This increase was greater with the macrophages from vaccinated animals than those from normal animals. The results support the hypothesis that H2O2 production by macrophages is involved in killing M. tuberculosis in vivo. PMID- 6271162 TI - Herpes simplex virus and eczematous skin. PMID- 6271161 TI - Enhanced monocyte and neutrophil cytotoxicity and normal cyclic nucleotide levels in severe psoriasis. AB - Eighteen patients with active psoriasis were investigated for antibody-dependent cell-mediated cytotoxicity (ADCC) mediated by monocytes and neutrophil leukocytes. Patients with extensive psoriatic lesions showed increased ADCC whereas patients with minimal psoriasis had normal monocyte and neutrophil function. After clinical remission the ADCC became normal. No stimulatory factors in psoriatic serum could be demonstrated. The increased monocyte and neutrophil cytotoxicity in severe psoriasis is not explained by altered cyclic nucleotide levels as cAMP and cGMP levels were normal in psoriatic monocytes and neutrophils showing both increased and normal ADCC. Our results indicate that increased ADCC is secondary to the psoriatic activity. PMID- 6271163 TI - Morphology and kinetics of erythropoiesis in haemoglobin H disease. AB - Some aspects of erythroblast function have been investigated in two Thai patients with haemoglobin H disease. Electron microscope and high resolution autoradiographic studies revealed that (1) some erythroblasts and marrow reticulocytes contained branching intracytoplasmic inclusions and (2) the presence of such inclusions did not necessarily prevent the entry of cells either from the G1 phase into the S phase or from the G2 phase into mitosis. The distribution of early polychromatic erythroblasts in the different stages of the cell cycle was determined by the technique of combined Feulgen microspectrophotometry and 3H-thymidine autoradiography and found to be essentially normal. It is concluded that although the presence of an excess of alpha-chains in the beta-thalassaemia syndromes has been previously shown to be associated with a gross impairment of erythroblast proliferation, the presence of an excess beta-chains in HbH disease is not. PMID- 6271164 TI - Chlamydia trachomatis and herpes simplex virus IgA antibodies in cervical secretions of patients with cervical atypia. AB - The association of Chlamydia trachomatis (CT) and herpes simplex virus (HSV) with malignant or premalignant changes in the cervix uteri was studied by determining immunoglobulin A (IgA) antibodies in the cervical secretions of 28 women with inflammatory, 28 with dysplastic, 7 with malignant changes of the uterine cervix, and 26 healthy controls. In cervical secretions IgA antibodies to CT were found in 24 of 35 (69%) patients with malignant or premalignant changes, in 11 of 28 (39%) with cervicitis and in 3 of 26 (12%) controls. IgA antibodies to HSV were found in 10 of 35 (28%) patients with malignant atypic or dysplasia but in none of the women with cervicitis or the controls. The highest frequency of antibodies was found in the patients with cervical carcinoma. Serum IgA antibodies to CT and HSV were found equally on the patients and the controls. Our results suggest that in patients with cervical atypic, local IgA CT antibody production occurs. Whether this association is aetiological or coincidental can not be concluded from this study. PMID- 6271165 TI - Carcinofibroma--a variant of the mixed Mullerian tumour. Case report. AB - An endometrial neoplasm composed of both stromal and epithelial elements, in which only the epithelial element appeared malignant, is reported. Only two similar cases have previously been described. In view of the rarity of these neoplasms, little is known about their behaviour and this presents problems of management particularly in a young patient. PMID- 6271166 TI - A comparison of the efficiency of human embryo intestine, Hep 2 cells, and human embryo kidney cells for the primary isolation of ophthalmic viruses. AB - A comparison has been made of the efficiency of 3 cell systems, human embryo kidney (HEK), Hep 2, and a continuous line of human embryo intestine (HEI), for the isolation of ophthalmic viruses. A total of 40 herpes simplex, 51 adenoviruses, and 2 vaccinia viruses were isolated from 323 specimens. HEK cells were found to be the optimal system, 85 out of 93 (91%) of the viruses being detected in these cells alone. However, HEK cells are difficult to obtain, and therefore the use of a combination of the continuous cell lines HEI and Hep 2 is recommended as an alternative. 89% of the viruses were detected by this combination. The use of either HEI or Hep 2 cells alone was unsatisfactory. PMID- 6271167 TI - Purification of the two major forms of rat pituitary corticotropin using only reversed-phase liquid chromatography. PMID- 6271168 TI - Isolation and characterization of corticotropin- and melanotropin-related peptides from the neurointermediary lobe of the rat pituitary by reversed-phase liquid chromatography. AB - A novel procedure utilizing reversed-phase high-performance liquid chromatography for the extraction and purification of peptides from biological tissues has been applied to the isolation of corticotropin-like intermediary lobe peptide (CLIP) and alpha-melanocyte-stimulating hormone (alpha-MSH) from the neurointermediary lobe of the rat pituitary. The isolation and characterization of two major forms of CLIP and two major forms of alpha-MSH are described. The isolated peptides have been identified by using enzymatic digestions and peptide mapping. The main form of ClIP is a peptide which has been modified by phosphorylation of the serine residue at position 31. This is the first peptide of endocrine origin reported to be modified in such a manner. A non-phosphorylated form of CLIP was also present at lower concentrations. The main form of alpha-MSH was found to be N,-O-diacetyl-alpha-MSH, with the more familiar mono-N-acetyl-alpha-MSH present to a much smaller extent. Thus, in the rat neurointermediary lobe, the two main corticotropin-related peptides present are mostly in modified forms which are the result of posttranslational modifications. It is only by the use of methodology such as that described in this paper that small alterations in peptide structure may be identified. PMID- 6271169 TI - Intracellular adenosine 3',5'-phosphate binding proteins in Dictyostelium discoideum: partial purification and characterization in aggregation competent cells. AB - Three adenosine 3',5'-phosphate (cAMP) binding proteins were separated and partially purified from cytoplasmic extracts of Dictyostelium discoideum cells developed to aggregation competence. Two species, A and B, representing respectively 50% and 20% of the total activity, bind cAMP with very rapid kinetics and high specificity. Species A (Kd = 7.5 nM) is a monomeric protein of 36 000 daltons with a sedimentation coefficient of 2.3 S. Species B, which binds cAMP with positive cooperativity, also displays a high affinity for the ligand (Kd = 3.2 nM). This protein is present in the extracts as an equilibrium between monomeric, dimeric, and tetrameric forms with respective sedimentation coefficients of 2.4, 4.5, and 6.5 S; binding of cAMP to the monomer induces the appearance of the multimeric forms. A third cAMP binding protein (species C, Kd - 9.5 nM) was characterized as a larger protein (Mr 190 000, sedimentation coefficient of 9.2 S) which also binds adenosine and adenosyl derivatives. Species C represents 30% of the activity in the extracts and resemble the "adenosine analogue binding proteins" described in mammalian cells. The relevance of the properties of these proteins to the developmental process of D. discoideum amoebas is discussed. PMID- 6271171 TI - Isolation and characterization of the amino and carboxyl proximal fragments of the adenosine cyclic 3' ,5'-phosphate receptor protein of Escherichia coli. AB - The cyclic AMP receptor protein (CRP) is a positive and negative regulatory protein for gene expression in Escherichia coli. The protein has been cleaved proteolytically to determine the relation between CRP structure and function. In the presence of sodium dodecyl sulfate (NaDodSO4), chymotrypsin dissects CRP into two stable fragments of molecular weight 9500 (9.5K) and 13 000 (13K). After removal of NaDodSO4, the two fragments are resolved by Bio-Rex 70 chromatography in 6 M urea. Analyses of the terminal amino acids released from each fragment and cyanogen bromide cleavage products indicate that the 9.5K fragment is amino proximal in CRP while the 13K fragment is carboxyl proximal. Notable features of amino acid composition are the relatively high amount of arginine and methionine in the 13K fragment and the retention in the 9.5K fragment of the two tryptophans present in the CRP subunit. Following isoelectric focusing in 8 M urea, the 9.5K fragment, 22.5K CRP, and 13K fragment migrate to pH 5.5, 8.3, and 10.3, respectively. While CRP is a cAMP-stimulated DNA binding protein, the 13K fragment binds to DNA in the presence and absence of cAMP. The 9.5K fragment associates to form dimers and decamers. These data are consonant with a model in which the DNA binding domain is present in the carboxyl proximal region of CRP while the amino proximal region contains the subunit-subunit interaction sites and much of the cAMP binding domain. PMID- 6271170 TI - Thermodynamic and kinetic examination of protein stabilization by glycerol. AB - The effect of concentrated glycerol on the thermal transitions of chymotrypsinogen and ribonuclease has been examined by differential spectrophotometry at 293 and 287 mm, respectively. It was found that for both proteins addition of glycerol raises the transition temperature, the increase in Tm being greater for ribonuclease than for chymotrypsinogen. This increase in the free energy of denaturation appears to reflect primarily a decrease in the entropy change. Analysis in terms of the Wyman linkage equation shows that, for both proteins, the exclusion of glycerol from the protein domain increases on denaturation i.e., the chemical potential of glycerol becomes even more positive when the protein unfolds relative to the native structure. This provides the thermodynamic stabilization free energy. Results of the kinetic examination of the slow unfolding reaction are consistent with the concept that the preferential exclusion of glycerol is related, at least in part, to enhanced solvent ordering. PMID- 6271172 TI - Modification of bovine pancreatic ribonuclease A with the site-specific reagent 4 arsono-2-nitrofluorobenzene. Spectrophotometric titration of arsononitrophenyl ribonuclease A derivatives. AB - The 4-arsono-2-nitrophenyl chromophore can serve as a versatile spectrophotometric probe of the surface structure of proteins. Values of pK1' and pK2' for the arsonic acid ionizations are near 3 and 8, respectively, and the presence of nearby positive and negative charges produces substantial alterations in the spectral response of the probe. Changes in the extinction at the wavelength of maximum difference are 30-50% of the extinction coefficients, epsilonmax, for each ionization of the arsonic acid moiety. The titration of 41 (4-arsono-2-nitrophenyl)ribonuclease A indicates that the arsonate dianion binds near the active-site histidine residues. With protonation of a carboxylate side chain in the acidic region, presumably aspartic acid-121, the active site is disrupted. The 41-(4-arsono-2-nitrophenyl) group interacts to a greater degree with the histidine-119 side chain than it does with the histidine-12 residue. Interactions of uridine or 3'-cytidylic acid with the ligand-binding region of 41 (4-arsono-2-nitrophenyl) ribonuclease A modify the spectrophotometric response extensively. 3'-Cytidylic acid binds 41-(4-arsono-2-nitrophenyl) ribonuclease A with an affinity 300 times less than that for native ribonuclease A and 17 times lower than that for 41-(2,4-dinitrophenyl) ribonuclease A. The arsononitrophenyl chromophore is responsive to changes in the active site of ribonuclease A induced by such perturbants as ligand binding, chemical modification, and both acid and thermal denaturation. PMID- 6271173 TI - Regulation of rat liver phosphoenolpyruvate carboxykinase (GTP) messenger ribonucleic acid activity by N6, O2'-dibutyryladenosine 3',5'-phosphate. AB - N6,O2'-Dibutyryladenosine 3',5'-phosphate (Bt2cAMP) induces the synthesis of the gluconeogenic enzyme, phosphoenolpyruvate carboxykinase (GTP) (EC 4.1.32), in rat liver by increasing the activity of messenger ribonucleic acid (mRNA) coding for this enzyme (mRNAPEPCK) more than 20-fold (from less than 0.01% to greater than 0.20% of total mRNA activity) as determined by using in vitro translation systems which measure only active mRNAPEPCK. The increase in mRNAPEPCK activity could result from increased synthesis, increased processing, or decreased inactivation rates. Actinomycin D and cordycepin inhibit mRNAPEPCK induction by 89% and 70%, respectively, a result that indicates a requirement for ongoing RNA synthesis but that does not distinguish which of these steps is regulated by cAMP. We have employed a kinetic approach, not involving RNA synthesis inhibitors, to determine the half-life of mRNAPEPCK both during a period of deinduction following glucose feeding and during a subsequent induction by Bt2cAMP. An estimated half-life of 20 +/-5 min during both of these periods indicates that Bt2cAMP has no effect on the rate of inactivation of mRNAPEPCK. We conclude that Bt2cAMP effects the increase in activity of mRNAPEPCK by promoting its synthesis or processing. PMID- 6271174 TI - T7 ribonucleic acid polymerase-promotor interactions. PMID- 6271175 TI - Immobilization of a spin-labeled fatty acid chain covalently attached to Ca2+ ATPase from sarcoplasmic reticulum suggests an oligomeric structure. PMID- 6271176 TI - Titration of the phase transition of phosphatidylserine bilayer membranes. Effects of pH, surface electrostatics, ion binding, and head-group hydration. AB - The dependence of the gel-to-fluid phase transition temperature of dimyristoyl- and dipalmitoylphosphatidylserine bilayers on pH, NaCl concentration, and degree of hydration has been studied with differential scanning calorimetry and with spin-labels. On protonation of the carboxyl group (pK2app = 5.5), the transition temperature increases from 36 to 44 degrees C in the fully hydrated state of dimyristoylphosphatidylserine (from 54 to 62 degrees C for dipalmitoylphosphatidylserine), at ionic strength J = 0.1. In addition, at least two less hydrated states, differing progressively by 1 H2O/PS, are observed at low pH with transition temperatures of 48 and 52 degrees C for dimyristoyl- and 65 and 68.5 degrees C for dipalmitoylphosphatidylserine. On deprotonation of the amino group (pK3app = 11.55) the transition temperature decreases to approximately 15 degrees C for dimyristoyl- and 32 degrees C for dipalmitoylphosphatidylserine, and a pretransition is observed at approximately 6 degrees C (dimyristoylphosphatidylserine) and 21.5 degrees C (dipalmitoylphosphatidylserine), at J = 0.1. No titration of the transition is observed for the fully hydrated phosphate group down to pH less than or equal to 0.5, but it affinity for water binding decreases steeply at pH greater than or equal to 2.6. Increasing the NaCl concentration from 0.1 to 2.0 M increases the transition temperature of dimyristoyphosphatidylserine by approximately 8 degrees C at pH 7, by approximately 5 degrees at pH 13, and by approximately 0 degrees C at pH 1. These increases are attributed to the screening of the electrostatic titration-induced shifts in transition temperature. On a further increase of the NaCl concentration to 5.5 M, the transition temperature increases by an additional 9 degree C at pH 7, 13 degree C at pH 13, approximately 7 degree C in the fully hydrated state at pH 1, and approximately 4 and approximately 0 degree C in the two less hydrated states. These shifts are attributed to displacement of water of hydration by ion binding. From the salt dependence it is deduced that the transition temperature shift at the carboxyl titration can be accounted for completely by the surface charge and change in hydration of approximately 1 H2O/lipid, whereas that of the amino group titration arises mostly from other sources, probably hydrogen bonding. The shifts in pK (delta pK2 = 2.85, delta pK3 = 1.56) are consistent with a reduced polarity in the head-group region, corresponding to an effective dielectric constant epsilon approximately or equal to 30, together with surface potentials of psi congruent to -100 and -150 mV at the carboxyl and amino group pKs, respectively. The transition temperature of dimyristoylphosphatidylserine-water mixtures decreases by approximately 4 degree C each water/lipid molecule added, reaching a limiting value at a water content of approximately 9-10 H2O/lipid molecule. PMID- 6271177 TI - Linear relations between proton current and pH gradient in bacteriorhodopsin liposomes. AB - The dependence of proton movement across the membrane of bacteriorhodopsin liposomes on the pH gradient was investigated. Under the appropriate experimental conditions, endogenous proton (or hydroxyl) leakage, proton movement catalyzed by protonophore or nigericin, and light-driven proton translocation depend linearly on the pH gradient. This justifies the use of linear proton flux vs. protonmotive force relations in a recent mosaic thermodynamic description of ion translocation in bacteriorhodopsin liposomes [Westerhoff, H. V., Scholte, B. J., & Hellingwerf, K. J. (1979) Biochim. Biophys. Acta 547, 544-560]. Since bacteriorhodopsin liposomes are a model system for all biological energy transducing systems in which proton pumps are involved, these findings also explain linear relations between proton flux and protonmotive force observed in and postulated for those systems. In cases where the membrane potential is not clamped at a low value, an initial phase of rapid proton movement occurs, followed by a phase of slower proton movement. The rate of proton movement during the slow phase is again linear with the pH gradient. Such a linear relation is not observed for the fast phase. Since the rapid proton movement phase is also observed in liposomes without bacteriorhodopsin, it is not due (only) to dissociation of scalar protons from bacteriorhodopsin. We suggest that during the initial phase of proton movement, the proton flux is not yet electrically compensated by the fluxes of other ions. PMID- 6271178 TI - Reactions of nitric oxide with tree and fungal laccase. AB - The reactions of nitric oxide (NO) with the oxidized and reduced forms of fungal and tree laccase, as well as with tree laccase depleted in type 2 copper, are reported. The products of the reactions were determined by NMR and mass spectroscopy, whereas the oxidation states of the enzymes were monitored by EPR and optical spectroscopy. All three copper sites in fungal laccase are reduced by NO. In addition, NO forms a specific complex with the reduced type 2 copper. NO similarly reduces all of the copper sites in tree laccase, but it also oxidizes the reduced sites produced by ascorbate or NO reduction. A catalytic cycle is set up in which N2O, NO2-, and various forms of the enzyme are produced. On freezing of fully reduced tree laccase in the presence of NO, the type 1 copper becomes reoxidized. This reaction does not occur with the enzyme depleted in type 2 copper, suggesting that it involves intramolecular electron transfer from the type 1 copper to NO bound to the type 2 copper. When the half-oxidized tree laccase is formed in the presence of NO, a population of molecules exists which exhibits a type 3 EPR signal. A triplet EPR signal is also seen in the same preparation and is attributed to a population of the enzyme molecules in which NO is bound to the reduced copper of a half-oxidized type 3 copper site. PMID- 6271179 TI - Valyl-tRNA synthetase from baker's yeast. Ligand binding properties and stability of the enzyme-bound adenylate. PMID- 6271181 TI - Reliability of nitroxide spin probes in reporting membrane properties: a comparison of nitroxide- and deuterium-labeled steroids. AB - The reliability for the study of membrane properties of the steroid nitroxide spin probe, 3-doxylcholestane, was tested by comparison of analogous data for the deuterated steroid, cholesterol-3 alpha-d. Good agreement between the two probes was found for the dependence of their order parameters on variation of temperature or cholesterol concentration in egg phosphatidylcholine bilayers. This finding is contrasted with the results of a previous study of fatty acid probes where poor agreement was found for the spectral responses of nitroxide- and deuterium-labeled species. The angular dependence of the ESR spectra of nitroxide-labeled probes in oriented multibilayer films was examined to determine if the probes were oriented in a tilted fashion in the bilayer. The 3 doxylcholestane probe and a doxylstearic acid labeled at position 14 orient with their long molecular axes perpendicular to the bilayer plane. In contrast, the stearic acid probe nitroxide labeled at position 5 does not appear to orient in such a fashion. We suggest that the behavior of the latter probe reflects the difficulty of inserting a bulky nitroxide group into a highly ordered region of the bilayer rather than an inherent tilting of the phospholipid acyl chains. On the basis of the comparisons between various types of probes, some suggestions are made concerning the choice of ESR spin probe to obtain reliable information in membrane studies. PMID- 6271180 TI - 5'-Nucleotidase from rat heart. AB - 5'-Nucleotidase has been extracted from rat heart and purified to apparent homogeneity. The enzyme is a glycoprotein. Gel electrophoresis in the presence of sodium dodecyl sulfate indicates that the apparent molecular weight of the subunit is 74 000 at several different gel concentrations. Cross-linking of the native enzyme with dimethylpimelimidate followed by gel electrophoresis shows that the enzyme is a dimer. The enzyme hydrolyzes all nucleoside 5' monophosphates tested. A comparison of Vmax/Km for 14 different substrates shows that AMP is the best substrate. The enzyme shows lowest Km values for AMPS, AMP, isoAMP, GMP, and IMP. It shows no activity with nucleoside 2'- and 3' monophosphates, sugar phosphates, and p-nitrophenyl phosphate, even when tested at high enzyme concentrations. The optimum activity of the enzyme occurs at pH 7.5 with AMP as substrate. Above this pH, buffer ions affect the activity in a complex manner, a second optimum being observed under some conditions. Magnesium ions activate the enzyme above pH 7.5 in the presence of some buffer ions but not of others. Magnesium ions show only a slight activation when the reaction is run in diethanolamine buffer, pH 9.5, at 30 degrees C; the activation in this buffer is considerably greater when the reaction is run at 37 degrees C. The enzyme is strongly inhibited by free ADP, maximum inhibition occurring below pH 6. The ADP inhibition is diminished as the pH is raised above 6, becoming negligible above pH9. The enzyme is inhibited by EDTA. The inhibition is partially reversed when the EDTA is removed from the enzyme by gel filtration. This as well as other evidence indicates that the enzyme contains a tightly bound metal ion. PMID- 6271182 TI - Lipid--protein multiple binding equilibria in membranes. AB - Phospholipids at the lipid--protein interface of membrane proteins are in dynamic equilibrium with fluid bilayer. In order to express the number of binding sites (N) and the relative binding constants (K) in terms of measurable quantities, the equilibrium is formulated as an exchange reaction between lipid molecules competing for hydrophobic sites on the protein surface. Experimental data are reported on two integral membrane proteins, cytochrome oxidase and (Na,-K) ATPase, reconstituted into defined phospholipids. Electron spin resonance measurements on reconstituted preparations of beef heart cytochrome oxidase in 1,2-dioleoyl-sn-3-phosphatidylcholine containing small quantities of the spin labeled phospholipid 1-palmitoyl-2-(14-proxylstearoyl)-sn-3-phosphatidylcholine (PC*) gave a linear plot of bilayer/bound PC* vs. the lipid/protein ratio as predicted by the theory, with K congruent to 1 and N = 40 (normalized to heme aa3). This demonstrates that the spin-label moiety attached to the hydrocarbon chain does not significantly perturb the binding equilibria. In the second experimental system, (Na,K)-ATPase purified from rectal glands of Squalus acanthias was reconstituted with defined phosphatidylcholines as the lipid solvent and spin-labeled phospholipids with choline or serine head groups (PC*, PS*) as the solute. The (Na,K)-ATPase has a larger number of lipid binding or contact sites (N = 60-65 per alpha 2 beta 2 dimer) and exhibits a detectably larger average binding constant for the negatively charged phosphatidylserine than for the corresponding phosphatidylcholine. These results show that a multiple equilibria, noninteracting site binding treatment can account for the behavior of lipids exchanging between the protein surface and the lipid bilayer. Selective sites among a background of nonselective sites are experimentally detectable as a change in the measured relative binding constant. PMID- 6271183 TI - Lipid dependence of glucose-6-phosphate phosphohydrolase: a study with purified phospholipid transfer proteins and phosphatidylinositol-specific phospholipase C. PMID- 6271184 TI - Interaction of hemagglutinating virus of Japan with erythrocytes as studied by release of a spin probe from virus. PMID- 6271185 TI - Synthesis and assembly of cytochrome c oxidase in synchronous cultures of yeast. AB - Yeast cells growing synchronously in glucose medium accumulate in the cytosol, the cytosolically made subunits of cytochrome oxidase, during the G1 and early-S phases. The mitochondrially made subunits, on the other hand, are detected only after the mid-S phase. The cytosolically synthesized subunits are integrated into the membrane after the mid-S phase. PMID- 6271186 TI - pH-induced conformational transitions of ferricytochrome c: a carbon-13 and deuterium nuclear magnetic resonance study. PMID- 6271187 TI - Guanidine hydrochloride induced unfolding of yeast iso-2 cytochrome c. AB - The properties of the guanidine hydrochloride induced unfolding transition of iso 2 cytochrome c (iso-2) from Saccharomyces cerevisiae have been investigated by using kinetic and equilibrium techniques and have been compared with previously published studies of horse cytochrome c, which differs from iso-2 by 46% in amino acid sequence. Measurements of absorbance in the ultraviolet and visible spectral regions as a function of guanidine hydrochloride concentration give superimposable equilibrium transition curves with a midpoint of 1.15 M at pH 7.2 and 20 degrees C. A two-state analysis of the equilibrium data gives a Gibbs free energy of unfolding of 3.1 kcal/mol at 20 degrees C in the absence of denaturant. This agrees well with the predicted difference in stability between S. cerevisiae iso-2 and horse cytochrome c estimated from the free energies of transfer of buried hydrophobic groups. Three kinetic phases associated with folding can be detected throughout most of the transition zone. Two of the phases are detected by stopped-flow mixing experiments. The third phase is over within the mixing time of the flow experiments but is detectable by temperature jumps. At 20 degrees C, pH 7.2, the slowest phase (T1) is in the 20-100-s time range, the middle phase (T2) is in the 0.1-3-s range, and the fastest phase (T3) is on the order of 1 ms. For the reactions observed in the stopped flow (T1 and T2), a simplified three-state mechanism can be used to predict quantitatively the relative amplitudes of the phases and the equilibrium unfolding curve from the observed time constant data. Previously this same mechanism has been successful in describing the folding reactions of horse cytochrome c [Hagerman, P. J. (1977) Biopolymers 16, 731]. We suggest that the qualitative features of protein folding reactions may be conserved among homologous proteins. PMID- 6271188 TI - Synthesis of nucleoside 3'-(S-alkyl phosphorothioates) and their use as substrates for nucleases. AB - The synthesis of cytidine, uridine, guanosine, and adenosine 3'-(S-methyl phosphorothioates) by treatment of the 2',5'-di-O-(4-methoxytetrahydropyran-4 yl)ribonucleosides with 2-(methylthio 4H-1,3,2-benzodioxaphosphorin 2-oxide is described. These nucleotide analogues are stable compounds both in the solid state and the neutral aqueous solution. All four of these compounds are degraded by RNase T2 to the parent nucleotides and methanethiol. In addition, cytidine and uridine 3'-(S-methyl phosphorothioates) are substrates for bovine pancreatic ribonuclease and guanosine 3'-(S-methyl phosphorothioate) is a substrate for RNase T1 and RNase U1. When used in conjunction with a chromophore-producing reagent, nucleoside 3'-(S-methyl phosphorothioates) provide a means for direct kinetic measurement of ribonuclease activity over a wide pH range (pH 2-9). The reactivities of these substrates with ribonucleases are compared to the reactivities of other synthetic substrates as well as a number of natural substrates. The utility of ribonucleoside 3'-(S-methyl phosphorothioates) as substrates for the assay of ribonucleases is discussed. PMID- 6271189 TI - Purification and some properties of a deoxyribonucleic acid endonuclease endogenous to rat liver chromatin. AB - A deoxyribonucleic acid (DNA) endonucleolytic activity has been purified from a 0.3 M KCl extract of rat liver chromatin by a combination of selective precipitation and ion-exchange and gel filtration chromatography. The purified protein has a molecular weight of 35 000 as determined by Sephadex G-200 gel filtration and sodium dodecyl sulfate-acrylamide gel electrophoresis. The nuclease activity is stimulated by the addition of Mg2+ and thus may represent the Mg2+-activated DNase endogenous to chromatin. The purified enzyme has the ability to make both single-strand nicks and double-strand cuts in DNA. PMID- 6271190 TI - Effect of amphotericin B on membranes: a spin probe study. AB - The effect of the polyene antibiotic amphotericin B on the electron paramagnetic resonance spectra of lipid probes intercalated in model membranes was examined. When the antibiotic was added to the aqueous phase, no spectral effects occurred. However, when the antibiotic was incorporated during membrane preparation, changes in spectral parameters suggested the appearance of a new phase. The spectral changes do not necessarily corroborate the pore models proposed previously for amphotericin B in membranes. With a spin probe that partitions between water and membrane, an interaction between the amphotericin B and probe is observed. This interaction does not occur in the membrane, but in the aqueous phase, between the probe and the aggregated antibiotic. Some of the equilibria involving the antibiotic appear to be achieved slowly. PMID- 6271191 TI - Isolation and partial characterization of membrane protein constituents of human neutrophil receptors for chemotactic formylmethionyl peptides. AB - Plasma membranes of human neutrophils were solubilized in buffer containing a nonionic detergent and applied to a formylmethionylleucylphenylalanine (fMet-Leu Phe)-Sepharose column that was washed and eluted with the chemotactic peptide fMet-Leu-Phe. Analysis of the eluate by filtration on Bio-Gel P150 in sodium dodecyl sulfate (NaDodSO4) buffer and by NaDodSO4-polyacrylamide gel electrophoresis revealed three predominant membrane proteins of approximate molecular weight 94 000 (MP-1), 68 000 (MP-2), and 40 000 (MP-3), of which MP-2 accounted for 74--93% of the total protein. Purified MP-1 and MP-2 contained an above average content of hydrophobic amino acids, while MP-2 and MP-3 had an above average content of acid and/or amide amino acids and a below average content of basic amino acids. MP-2 and MP-3, but not MP-1, bound [3H]fMet-Leu-Phe in equilibrium dialysis chambers. Both MP-2 and MP-3 exhibited high-affinity sites with a valence of 0.2--0.3 and mean KA values of 9 x 10(8) and 2 x 10(7) M 1, respectively, and low-affinity sites with a valence of 0.3--0.5 and mean KA values of 3 x 10(7) and 2 x 10(6) M-1 (n = 3). The specificity of the binding of fMet-Leu-Phe was suggested by the failure of MP-2 and MP-3 to bind lipid chemotactic factors and to adhere to a Sepaharose column to which had been coupled chemotactic fragments of the fifth component of complement. A series of synthetic formylmethionyl peptides exhibited the same rank order of potency as inhibitors of the binding of [3H]fMet-Leu-Phe by MP-2 and as stimuli of neutrophil chemotaxis. Membrane proteins isolated by fMet-Leu-Phe-Sepharose affinity chromatography may represent constituents of specific human neutrophil receptors for chemotactic peptides. PMID- 6271192 TI - Influence of ether linkage on the lamellar to hexagonal phase transition of ethanolamine phospholipids. PMID- 6271193 TI - Electron paramagnetic resonance studies and insulin-like effects of vanadium in rat adipocytes. PMID- 6271194 TI - Mechanisms of hydrolysis of adenosine 5'-triphosphate, adenosine 5'-diphosphate, and inorganic pyrophosphate in aqueous perchloric acid. AB - The acid-catalyzed hydrolysis of adenosine 5'-triphosphate (ATP) has been found to give rise both to adenosine 5'-diphosphate (ADP) and inorganic phosphate and to adenosine 5'-phosphate (AMP) and inorganic pyrophosphate. Kinetic and isotope studies on the mechanism of hydrolysis of ATP therefore depend on a knowledge of the mechanism of hydrolysis of the polyphosphate products, ADP and inorganic pyrophosphate. The latter reactions have been studied over the acidity range 1--5 M perchloric acid at 25 degrees C while the more complex problem of the hydrolysis of ATP has been followed at a single acidity (3 M perchloric acid). The positions of bond fission have been determined for both ATP and ADP. PMID- 6271195 TI - Spin-label studies of head-group specificity in the interaction of phospholipids with yeast cytochrome oxidase. PMID- 6271196 TI - The absorbance coefficient of beef heart cytochrome c1. AB - Isolated cytochrome c1 contains endogenous reducing equivalents. They can be removed by treating the protein with sodium dithionite followed by chromatography. This treatment has no effect on the reaction with cytochrome c, nor does it alter the optical spectrum, or the polypeptide or amino acid composition of the protein. Both the titration of dithionite-treated ferrocytochrome c1 with potassium ferricyanide and the anaerobic titration of dithionite-treated ferricytochrome c1 with NADH in the presence of phenazine methosulphate lead to the same value for the absorbance coefficient of cytochrome c1: 19.2 mM-1 . cm-1 at 552.4 nm for the reduced-minus-oxidised form. This value was also obtained when the haem content was determined by comparing the spectra of the reduced pyridine haemochromes of cytochrome c and cytochrome c1. Comparison of the optical spectra of cytochrome c and cytochrome c1 by integration shows equal transition moments for the transitions in the porphyrin systems of both proteins. A set of equations with which the concentration of the cytochromes aa3, b, c and c1 can be calculated from one reduced-minus-oxidised difference spectrum of a mixture of these proteins. PMID- 6271197 TI - Oxidation of sulphide by cytochrome aa3. AB - The effectiveness of H2S as an inhibitor of cytochrome c oxidase increase (Ki decreases) with sulphide concentration. A spectroscopic change in cytochrome aa3 is induced aerobically by sulphide at the same rate as that calculated for inhibition. The initial spectroscopic product is not inhibited, but an 'oxygenated' (oxyferri) form of the enzyme. Stoichiometric sulphide addition to cytochrome aa3 under anaerobic conditions produces another low-spin form of the enzyme; subsequent admission of oxygen gives rise to the 607 nm compound. At high enzyme levels sulphide itself acts as a substrate measured polarographically, with an oxygen uptake proportional to the amount of sulphide added. Binding of sulphide to ferric enzyme probably causes reduction at the oxygen-sensitive a3-Cu centre, which is followed aerobically by reoxidation to the oxyferri state via the 607 nm intermediate. A stable sulphide complex is formed only after the reduction of cytochrome a; but once formed this inhibited species is retained if cytochrome a is reoxidized. PMID- 6271198 TI - Inhibition of cytochrome c oxidase function by dicyclohexylcarbodiimide. AB - Dicyclohexylcarbodiimide (DCCD) reacted with beef heart cytochrome c oxidase in inhibit the proton-pumping function of this enzyme and to a lesser extent to inhibit electron transfer. The modification of cytochrome c oxidase in detergent dispersion or in vesicular membranes was in subunits II-IV. Labelling followed by fragmentation studies showed that there is one major site of modification in subunit III. DCCD was also incorporated into several sites in subunit II and at least one site of subunit IV. The major site in subunit III has a specificity for DCCD at least one order of magnitude greater than that of other sites (in subunits II and IV). Its modification could account for all of the observed effects of the reagent, at least for low concentrations of DCCD. Labelling of subunit II by DCCD was blocked by prior covalent attachment of arylazidocytochrome c, a cytochrome c derivative which binds to the high-affinity binding site for the substrate. The major site of DCCD binding in subunit III was sequenced. The label was found in glutamic acid 90 which is in a sequence of eight amino acids remarkably similar to the DCCD-binding site within the proteolipid protein of the mitochondrial ATP synthetase. PMID- 6271199 TI - Isolation and purification of the cytochrome oxidase of Azotobacter vinelandii. AB - A membrane-bound cytochrome oxidase for Azobacter vinelandii was purified 20-fold using a detergent-solubilization procedure. Activity was monitored using as ascorbate-TMPD oxidation assay. The oxidase was 'solubilized' from a sonic-type electron-transport particle (R3 fraction) using Triton X-100 and deoxycholate. Low detergent concentrations first solubilized the flavoprotein oxidoreductases, then higher concentrations of Triton X-100 and KCl solubilized the oxidase, which was precipitated at 27-70% (NH4)2SO4. The highly purified cytochrome oxidase has a V of 60-78 microgatom O consumed/min per mg protein. TMPD oxidation by the purified enzyme was inhibited by CO, KCN, NaN3 and NH2OH; NaNO2 (but not NaNO3) also had a potent inhibitory effect. Spectral analyses revealed two major hemoproteins, the c-type cytochrome c4 and cytochrome o; cytochromes a1 and d were not detected. The Azotobacter cytochrome oxidase is an integrated cytochrome c4-o complex, TMPD-dependent cytochrome oxidase activity being highest in preparations having a high c-type cytochrome content. This TMPD-dependent cytochrome oxidase serves as a major oxygen-activation site for the A. vinelandii respiratory chain. It appears functionally analogous to cytochrome a+a3 oxidase of mammalian mitochondria. PMID- 6271200 TI - Structural studies on the cytochrome c oxidase proton pump using a spin-label probe. AB - We report studies in which we have used N-(2,2,6,6-tetramethylpiperidyl-l-oxyl) N' -cyclohexylcarbodiimide, a spinlabel analogue of N,N' dicyclohexylcarbodiimide, to investigate the structural aspects of the cytochrome c oxidase proton pump. We establish that the spin label binds to the reconstituted enzyme at the same site as does N,N' -dicyclohexylcarbodiimide, i.e., within subunit III. ESR studies of the bound spin label indicate that its binding site is situated in an apolar region of the enzyme, though close to its surface. The binding of the spin label to the free oxidase is different form that with the reconstituted enzyme, leading to spin-spin exchange between the bound probe molecules. From this and the fact that N,N' -dicyclohexylcarbodiimide binds to subunits III and IV in the free oxidase, we conclude that these two subunits are at the most 20 A apart. PMID- 6271201 TI - The hydrophobic nature of the pig intrinsic factor receptor in the intestine. AB - The pig intestinal intrinsic factors receptor has been isolated and dissociated into its alpha and beta subunits. The beta subunit was found to be more hydrophobic than the alpha subunit. In a detergent solution only the alpha subunit was accessible to digestion with papain. The whole isolated receptor was introduced into artificial single bilayer liposomes where is apparently was randomly oriented. Liposomes containing the receptor were digested with papain and the polypeptide segments that stayed in the lipid fraction were extracted and analyzed using sodium dodecyl sulfate polyacrylamide gel electrophoresis. Four species were found with Mr values of 23 000, 45 000, 70 000 and 86 000. PMID- 6271202 TI - Properties of the large ion-permeable pores formed from protein F of Pseudomonas aeruginosa in lipid bilayer membranes. AB - The incorporation of porin protein F from the outer membrane of Pseudomonas aeruginosa into artificial lipid bilayers results in an increase of the membrane conductance by many orders of magnitude. The membrane conductance is caused by the formation of large ion-permeable channels with a single-channel conductance in the order of 5 nS for 1 M alkali chlorides. The conductance has an ohmic current vs. voltage relationship. Further information on the structure of the pore formed by protein F was obtained by determining the single-channel conductance for various species differing in charge and size, and from zero current potential measurements. The channel was found to be permeable for large organic ions (Tris+, N(C2H5)4+, Hepes-) and a channel diameter of 2.2 nm could be estimated from the conductance data (pore length of 7.5 nm). At neutral pH the pore is about two times more permeable for cations than for anions, possibly caused by negative charges in the pore. The consistent observation of large water filled pores formed by porin protein F in model membrane systems is discussed in the light of the known low permeability of the Ps. aeruginosa outer membrane towards antibiotics. It is suggested that this results from a relatively low proportion of open functional porin protein F pores in vivo. PMID- 6271203 TI - Evidence against Na+-pump mediation of Ca2+-activated K+ transport and diuretic sensitive (Na+/K+)-cotransport. PMID- 6271204 TI - In vitro phosphorylation of the red blood cell cytoskeleton complex by cyclic AMP dependent protein kinase from erythrocyte membrane. AB - Hydrosoluble proteins extracted from human erythrocyte ghosts by dialysis at low ionic strength and alkaline pH contain a cyclic AMP-dependent protein kinase which phosphorylates in vitro the cytoskeleton components in crude extracts. Spectrin components 1 and 2, actin and protein band 4.1 undergo this cyclic AMP dependent endogenous phosphorylation together with low molecular weight peptides solubilized with the cytoskeleton in hydrosoluble extract. However, pure spectrin and purified erythrocyte G-actin were not phosphorylated by purified cyclic AMP dependent protein kinase from erythrocyte membrane. Purified G-actin when added free to crude extract does not undergo phosphorylation by the cyclic AMP dependent protein kinase present in this extract. In contrast, purified cyclic AMP-dependent protein kinase added either to crude extract or to the purified cytoskeleton complex (spectrin, actin and protein band 4.1), phosphorylates spectrin, actin and protein band 4.1. We can conclude that (1) cyclic AMP dependent phosphorylation of red cell cytoskeleton occurs in vitro only when the cytoskeleton components are in a complexed form; (2) red cell actin, like other cellular actins, may be phosphorylated by cyclic AMP-dependent protein kinase but only in the oligomeric form and not in the G form. PMID- 6271205 TI - Sphingolipid-induced enhancement of receptor-mediated uptake of low density lipoproteins in normal and receptor-deficient human skin fibroblasts. AB - (1) The receptor mediated endocytosis of homologous LDL by human skin fibroblasts can be significantly enhanced by prior incubation of the cells with sphingolipids. Gangliosides GM1 or GD1a, their desialylated derivatives and sphingosine stimulate binding and uptake to LDL by up to 40% of normal values. The effect is observed in normal fibroblasts, LDL receptor deficient fibroblasts or in tunicamycin-treated cells with a reduced number of functional receptors but is dependent on the time of preincubation of the cells and the concentration of the sphingolipid in the medium. (2) Detailed studies on the ganglioside effect revealed, that cell bound gangliosides intensify the LDL-induced suppression of [14C] acetate incorporation into cholesterol. (3) The receptor dependence and relative receptor specificity of the sphingolipid effect is evident from the fact that (a) after complete suppression of receptor synthesis gangliosides fail to stimulate uptake of LDL, that (b) fatty acids or lipids not containing sphingosine are without effect and that (c) the receptor specific internalisation of alpha 2-macroglobulin or epidermal growth factor is not influenced by exogenous sphingolipids. PMID- 6271206 TI - A simple method for the isolation of basolateral plasma membrane vesicles from rat kidney cortex. Enzyme activities and some properties of glucose transport. AB - A procedure for preparing basolateral membrane vesicles from rat renal cortex was developed by differential centrifugation and Percoll density gradient centrifugation, and the uptake of D-[3H] glucose into these vesicles was studied by a rapid filtration technique. (Na+ + K+)-ATPase, the marker enzyme for basolateral membranes, was enriched 22-fold compared with that found in the homogenate. The rate of D-glucose uptake was almost unaffected by Na+ gradient (no overshoot). PMID- 6271207 TI - The effect of cytochrome oxidase on lipid chain dynamics. A nanosecond fluorescence depolarization study. AB - Molecular motions in membranes composed of purified cytochrome oxidase (EC 1.9.3.1) and synthetic lipid (L-alpha-dimyristoylphosphatidylcholine or L-alpha dioleoylphosphatidylcholine) at various ratios were investigated with a lipophilic fluorescent probe 1,6-diphenyl-1,3,5-hexatriene. Nanosecond fluorescence depolarization kinetics of the probe showed that the rod-shaped probe molecules perform a fast wobbling motion (restricted rotation) in all membranes studied, presumably reflecting the motion of lipid acyl chains. At temperatures where the pure lipid was in the liquid-crystalline phase, presence of cytochrome oxidase reduced the angular range of the wobbling motion, whereas its rate, the wobbling diffusion constant, was unaffected. On the other hand, incorporation of the protein into lipid in the gel phase resulted in the increase in the wobbling diffusion constant while the range of the wobbling motion remained the same. A time-dependent view of lipid dynamics that accounts for the above findings, as well as the results of recent electron spin resonance and nuclear spin resonance studies of protein-lipid interactions, is proposed. PMID- 6271208 TI - Effects of antidiuretic hormone on kinetic and energetic determinants of active sodium transport in frog skin. AB - The effects of antidiuretic hormone (ADH) on the rate of transepithelial active Na transport JaNa and the rate of suprabasal O2 consumption of Jsbr were studied in paired hemiskins of frog. Within some 30 min following administration of ADH both JaNa and Jsbr increased to near-maximal levels and then remained stable for at least an hour. On symmetric perturbation of the transepithelial electrical potential delta psi at 6-min intervals, the dependence of JaNa and Jsbr on delta psi was near-linear, both in control and experimental hemi-skins. The stability and near-linearity of the system permitted systematic analysis of the parameters of linear non-equilibrium thermodynamic (NET) and electrical equivalent circuit (EC) formulations. ADH (100 mU/ml) stimulated two of the three NET phenomenological L coefficients, as well as A, the affinity (negative Gibbs free energy) of a metabolic reaction driving transport. Observations at partially depressed levels of transport indicated that the effects of kinetic and energetic factors are to some extent discrete. EC analysis showed stimulation of the amiloride-sensitive conductance Ka, but not of the apparent electromitive force of Na transport 'ENa'. Similar effects were produced by 10 mU/ml of ADH or by 10 mM dibutyryl cyclic AMP, although less marked effects on the L coefficients were noted with the lower concentration of hormone. It is suggested that, in contrast to EC analysis, the NET formulation distinguishes between kinetic and energetic determinants of transport, supporting a dual mechanism of action of ADH. PMID- 6271209 TI - Electron paramagnetic resonance studies on the fluidity and surface dynamics of egg phosphatidylcholine vesicles containing gangliosides. AB - The influence of different gangliosides (GM1, GD1a, GT1b) on the fluidity and surface dynamics of phosphatidylcholine small unilamellar vesicles was studied by electron paramagnetic resonance. 5- and 16-nitroxystearic acid, sounding respectively the region close to the surface and that close to the hydrophobic core of the vesicle, were employed as spin-label probes. The signals released by 5-nitroxystearic acid showed that the presence of gangliosides reduced the mobility of the hydrocarbon chains around the probe. The effect increased by increasing ganglioside concentration, and diminished from GM1 to GD1a and GT1b. The decrease of membrane fluidity was also monitored by the 16-nitroxystearic acid probe. On addition of Ca2+ the fluidity of ganglioside-containing vesicles (as signalled by the 5-nitroxystearic acid probe) promptly decreased, therefore returning slowly to the original value. It is suggested that gangliosides cause strong side-side head group interactions on the bilayer surface--between ganglioside oligosaccharide chains and between ganglioside and phosphatidylcholine polar portions--which lead the lipid chains to assembly in a more rigid fashion. The influence of Ca2+ is interpreted as due to lateral phase separation in the vesicle membrane. This phenomenon can be related to the formation or stabilization of ganglioside clusters on the vesicle surface. PMID- 6271210 TI - The effects of membrane lipid order and cholesterol on the internal and external cationic sites of the Na+-K+ pump in erythrocytes. AB - cholesterol depletion alters the apparent affinity of the internal cationic sites and the maximal translocation rate but not the affinity of the external cationic sites of the Na+-K+ pump in human erythrocytes. To test whether these effects were mediated by a direct cholesterol-internal site interaction or by a change in membrane lipid order, the effects of five fluidizing amphiphiles (chlorpromazine, imipramine, benzyl alcohol, sodium oleate and sodium benzenesulphonate) on the kinetic parameters of the Na+-K+ pump were determined. The cholesterol removal and all the agents used induced dose-response decreases in membrane lipid order as measured by fluorescence polarization or ESR. Positive and neutral amphiphiles mimicked the effects of cholesterol removal on the affinity of the internal sites of the pump and to a lesser extent on the maximal translocation rate. Anionic amphiphiles had no effect on internal sites, probably because they distributed preferentially within the outer leaflet on the membrane. These results indicate that cholesterol controls the affinity of the internal sites of the Na+-K+ pump by altering the membrane lipid order. In contrast, neither cholesterol depletion nor the agents used altered the affinity of the external sites of the Na+-K+ pump. This difference in sensitivity to membrane lipids order suggests that internal and external cationic sites, although borne by the same protein, are in different lipid environments. PMID- 6271211 TI - The fraction of phosphatidylinositol that activates the (Na+ + K+)-ATPase in rabbit kidney microsomes is clearly associated with the enzyme protein. AB - 1. Extensive treatment of rabbit kidney microsomes with phosphatidylinositol specific phospholipase C under various conditions never resulted in more than 75% hydrolysis of the substrate. 2. The non-degraded fraction of the phosphatidylinositol (10-12 nmol per mg microsomal protein) could be recovered only by an acidic extraction procedure. 3. The (Na+ + K+)-ATPase activity found in those membranes was not affected by this treatment. 4. Complete degradation of phosphatidylinositol could be easily achieved when the phospholipase was applied to rat liver microsomes which do not contain any detectable (Na+ + K+)-ATPase activity. 5. It is concluded that in rabbit kidney microsomes a close association exists between the (Na+ + K+)-ATPase and that fraction of the phosphatidylinositol that is directly involved in the maintenance of its activity. PMID- 6271212 TI - A food dye, erythrosine B, increases membrane permeability to calcium and other ions. AB - A widely used food additive erythrosine B, which has been implicated in minimal brain dysfunction in children was examined for its ability to increase membrane permeability to calcium ions. Planar phospholipid bilayer membranes become permeable to calcium, potassium and chloride ions and when erythrosine B is added to the aqueous phase at concentrations which were used by others to demonstrate effects on neuromuscular preparations. The observed increase in permeability to Ca2+ was of sufficient magnitude that equivalent effects on cells would seriously tax the systems which maintain low cytoplasmic Ca2+ levels. The permeability increase in the lipid bilayer membrane is time dependent and increases with erythrosine B concentration raised to a high power (4 to 7). This indicates that the permeability pathway is generated by the cooperative action of a number of erythrosine molecules. This permeability increases dramatically with increasing transmembrane voltage indicating that cells or organelles bearing potentials across their membranes should be particularly sensitive to the dye. We propose that the neurological effects of erythrosine stem from the increased Ca2+ permeability. PMID- 6271213 TI - Biphasic effect of orthophosphate on the (Na, K)-pump of human red cells. AB - Orthophosphate (Pi) can both stimulate and inhibit the (Na, K)-pump in red cells. At concentrations below 0.5 mmol/l cells, Pi stimulated the pump, but at higher concentrations Pi was inhibitory. The stimulation was demonstrated in intact cells by preincubation with inosine (which leads to a reduction in cellular Pi concentration), and then by incubating cells in media with various Pi concentrations (which relieved the inhibition caused by inosine). In inosine treated cells there was an inverse relationship between the hematocrit during measurement of the fluxes and inhibition of the (Na, K)-pump; this was also a reflection of cellular Pi, which was lower in inosine-treated cels at low hematocrit. The stimulation of the (Na, K)-pump by Pi below 0.5 mmol/l cells was an indirect effect, due to synthesis of ATP by membrane-bound glycolytic enzymes, which required the appropriate substrates (in addition to Pi). This was shown by studies on inside-out vesicles made from red cell membranes. In the absence of the other substrates, Pi was inhibitory to Na transport in the vesicles. Above 0.5 mmol/l cells Pi was inhibitory to Na transport, both in inside-out vesicles and in intact cells. The mechanism of inhibition, probably a direct effect on the (Na, K)-pump, was not determined, though product inhibition seemed likely. The dependence on Pi of abnormal modes of Na transport by the pump (uncoupled Na efflux and Na/Na exchange) at low Pi concentrations was less than the dependence of normal Na/K exchange. This was attributed to a requirement by the abnormal modes of a lower rate of synthesis of ATP or a lower ATP concentration. PMID- 6271214 TI - Fluctuation analysis of Na+ channels modified by batrachotoxin in myelinated nerve. AB - (1) Single myelinated nerve fibers of Rana esculenta were treated with the steroidal alkaloid batrachotoxin, and Na+ currents and Na+-current fluctuations were measured near the resting potential under voltage-clamp conditions. Between test pulses the fibres were held at hyperpolarizing membrane potentials. (2) The spectral density of Na+-current fluctuations was fitted by the sum of a 1/f component and a Lorentzian function. The time constant tau c = 1/(2 pi fc) obtained from the corner frequency fc of the Lorentzian function approximately agreed with the activation time constant tau m of the macroscopic currents. (3) The conductance gamma of a single Na+ channel modified by batrachotoxin was calculated from the integral of the Lorentzian function and the steady-state Na+ current. At the resting potential V = 0 we obtained gamma - 1.6 pS, higher gamma values of 3.2 and 3.45 pS were found at V = --8 and --16 mV, respectively. (4) The conductance of a modified Na+ channel is significantly lower than the values 6.4 to 8.85 pS reported in the literature for normal Na+ channels. Hence, our experiments are in agreement with the view that batrachotoxin acts in an 'all-or none' manner on Na+ channels and creates a distinct population of modified channels. PMID- 6271215 TI - Evidence for convertible forms of soluble uterine cyclic nucleotide phosphodiesterase. AB - The cyclic nucleotide phosphodiesterase (3':5'-cyclic nucleotide 5' nucleotidohydrolase, EC 3.1.4.17) systems of many tissues show multiple physical and kinetic forms. In contrast, the soluble rat uterine phosphodiesterase exists as a single enzyme form with non-linear Lineweaver-Burk kinetics for cyclic AMP (app. Km of approx. 3 and 20 microM) and linear kinetics for cyclic GMP (app. Km of approx. 3 microM) since the two hydrolytic activities are not separated by a variety of techniques. In uterine cytosolic fractions, cyclic AMP is a non competitive inhibitor of cyclic GMP hydrolysis (Ki approx. 32 microM). Also, cyclic GMP is a non-competitive inhibitor of cyclic AMP hydrolysis (Ki approx 16 microM) at low cyclic GMP/cyclic AMP substrate ratios. However, cyclic GMP acts as a competitive inhibitor of cyclic AMP phosphodiesterase (Ki approx 34 microM) at high cyclic GMP/cyclic AMP substrate ratios. When a single hydrolytic form of uterine phosphodiesterase, separated initially by DEAE anion-exchange chromatography, is treated with trypsin (0.5 microgram/ml for 2 min) and rechromatographed on DEAE-Sephacel, two major forms of phosphodiesterase are revealed. One form elutes at 0.3 M NaOAc- and displays anomalous kinetics for cyclic AMP hydrolysis (app. Km of 2 and 20 microM) and linear kinetics for cyclic GMP (app. Km approx. 5 microM), kinetic profiles which are similar to those of the uterine cytosolic preparations. A second form of phosphodiesterase elutes at 0.6 M NaOAc- and displays a higher apparent affinity for cyclic AMP (app. Km approx. 1.5 mu) without appreciable cyclic GMP hydrolytic activity. These data provide kinetic and structural evidence that uterine phosphodiesterase contains distinct catalytic sites for cyclic AMP and cyclic GMP. Moreover, they provide further documentation that the multiple forms of cyclic nucleotide phosphodiesterase in mammalian tissues may be conversions from a single enzyme species. PMID- 6271216 TI - Regulatory properties of lysine-sensitive aspartokinase under equilibrium conditions. AB - The regulatory properties of the lysine-sensitive aspartokinase (ATP : L aspartate 4-phosphotransferase, EC 2.7.2.4) have been studied under equilibrium conditions by determining the effects of modifiers on the rate of equilibrium isotope exchange between ADP and ATP. The extent of inhibition by lysine, leucine or phenylalanine is almost independent of substrate concentration but is influenced by the substrate/product ratio. Inhibition by a given concentration of inhibitor is increased when the ADP/ATP ratio is increased indicating a regulatory interaction between end products and cellular energy metabolism. Lysine inhibition is cooperative under equilibrium conditions and the parameters of the Hill equation are nearly identical to those obtained in initial velocity studies. A cooperative heterotropic interaction between lysine and leucine is also observed by the ATP-ADP exchange assay just as it is in initial velocity assays. Thus, the regulatory features of aspartokinase that are observed in initial velocity studies are also manifest under equilibrium conditions as revealed by equilibrium isotope exchange rates. PMID- 6271217 TI - Dissociation and reassociation of prolyl 4-hydroxylase subunits after cross linking of monomers. AB - 1. Incubation of prolyl 4-hydroxylase (prolyl-glycyl-peptide, 2-oxoglutarate : oxygen oxidoreductase (4-hydroxylating), EC 1.14.11.2) with H2O2 leads to a decrease of 50% in the specific activity of enzyme tetramers, followed by dissociation into inactive dimers in which the monomers are covalently cross linked by S-S bridge formation. 2. Incubation of the enzyme with K3Fe(CN)6 leads to a comparable decrease in activity of enzyme tetramers. Addition of urea leads to dissociation into inactive dimers with similarly cross-linked monomers. 3. Removal of the dissociating agent leads to reassociation of cross-linked dimers to tetramers and to about 50% reactivation. The enzyme is further reactivated by preincubation with dithiothreitol. 4. Dissociation of the enzyme with dithiothreitol, urea or LiCl, or at low pH (4.15) produces inactive monomers, which could not be reassociated. PMID- 6271218 TI - Properties of a collagenolytic enzyme from Bipalium kewense. AB - A collagenolytic enzyme from the land planarian Bipalium kewense has been purified by preparative isoelectric focusing. The enzyme has a molecular weight of 47,000 +/- 2,000 and appears to be dimeric. It has an isoelectric point of 4.6 +/- 0.1 and a high content of acidic amino acids. The amino acid composition of the Bipalium collagenase is similar to that of human skin fibroblast collagenases but clearly different from previously reported collagenolytic proteases from other invertebrates, Uca pugilator and Hypoderma lineatum. In its action on guinea-pig collagen, the enzyme produces distinct products, at low incubation temperatures, different from those produced by vertebrate and other invertebrate collagenolytic enzymes. These products have glycine as their N-terminal amino acids. As determined by viscosity measurements, the Bipalium collagenase is more active on invertebrate, earthworm, collagen than it is on the vertebrate, Type I guinea-pig skin, collagen. The Bipalium collagenase differs from both bacterial and vertebrate collagenases as well as from invertebrate, collagenolytic serine proteases. PMID- 6271219 TI - Spectral properties of myeloperoxidase and its ligand complexes. AB - The effects of ligands with various field strengths on the optical absorption spectrum of myeloperoxidase have been investigated. As is the case with other hemoproteins, the Soret peak in the optical absorption spectra at 77 K moves to longer wavelengths when strong-field ligands are present, whereas binding of such ligands as chloride and fluoride, which stabilize the high-spin state, shows the opposite effect. With a ligand of intermediate field strength, such as azide, the optical spectrum is not affected at room temperature, but lowering of the temperature results in the formation of the low-spin form of the enzyme. Similarly, in native myeloperoxidase a spin state equilibrium is found in which the low-spin state is favoured at high ionic strength and displays corresponding changes in the optical spectra. From the ligand- and the temperature-induced changes in the optical spectra of the ferric enzyme it is concluded that the band at 620-630 nm is an alpha band of the low-spin heme iron species, whereas the bands at 500 and 690 nm are probably 'charge-transfer' bands of the heme with the iron in the high-spin state. PMID- 6271220 TI - An alkaline metallo-proteinase in the human uterine cervix an changes in its activity by cervical ripening. AB - Human uterine cervix at term pregnancy was found to contain an alkaline metallo proteinase by use of a synthetic substrate, 2,4-dinitrophenyl-L-Pro-L-Gln-Gly-L Ile-L-Ala-Gly-L-Gln-D-Arg. The enzyme (with a molecular weight of 3.8 . 10(4)) was most active around pH 9.2 toward casein and N alpha-benzoyl-DL-Arg-rho nitroanilide. [14C]-Gelatin and proteoglycan subunit were also substrates for the enzyme, but [14C]collagen was not. In particular, the enzyme digested gelatin 70 times faster than the novel neutral proteinase in the cervix. Although EDTA was a potent inhibitor, 1,10-phenanthroline, human serum, diisopropylfluorophosphate and elastatinal had no effect on the enzyme. Alkaline proteinase in term pregnant cervices was significantly higher than in non-pregnant ones. PMID- 6271221 TI - Subcellular localization and properties of pyridoxal phosphate phosphatases of human polymorphonuclear leukocytes and their relationship to acid and alkaline phosphatase. AB - Using a novel fluorimetric assay for pyridoxal phosphate phosphatase, human polymorphonuclear leucocytes were found to exhibit both acid an alkaline activities. The neutrophils were homogenised in isotonic sucrose and subjected to analytical subcellular fractionation by sucrose density gradient centrigfugation. The alkaline pyridoxal phosphate phosphatase showed a very similar distribution to alkaline phosphatase an was located solely to the phosphasome granules. Fractionation experiments on neutrophils treated with isotonic sucrose containing digitonin and inhibitor studies with diazotised sulphanilic acid and levamisole further confirmed that both enzyme activities had similar locations and properties. Acid pyridoxal phosphate phosphatase activity was located primarily to the tertiary granule with a partial azurophil distribution. Fractionation studies on neutrophils homogenised in isotonic sucrose containing digitonin and specific inhibitor studies showed that acid pyridoxal phosphate phosphatase and acid phosphatase were not the result of a single enzyme activity, Neutrophils were isolated from control subjects, patients with chronic granulocytic leukaemia and patients in the third trimester of pregnancy. The specific activities (munits/mg protein) of alkaline pyridoxal phosphate phosphatase an alkaline phosphatase varied widely in the three groups and the alterations occurred in a parallel manner. The specific activities of acid pyridoxal phosphate phosphatase and of acid phosphatase were similar in the three groups. These results, together with the fractionation experiments and inhibition studies strongly suggest that pyridoxal phosphate is a physiological substrate for neutrophil alkaline phosphatase. PMID- 6271222 TI - Iron-sulphur clusters in fumarate reductase from Vibrio succinogenes. AB - (1) The fumarate reductase complex from Vibrio succinogenes contains one FAD molecule, one [4Fe-4S]3+(3+,2+) and one [2Fe-2S]2+(2+,1+) cluster per enzyme molecule. Both clusters can be partly reduced by succinate. In the presence of excess Na2S2O4 and fumarate, the [2Fe-2S] cluster is completely oxidized, whereas the other cluster is largely reduced. (2) The [2Fe-2S] cluster is localized in the Mr, 31,000 subunit. The EPR spectrum of the reduced cluster in the isolated subunit differs slightly in line width, but not in g-value, from the spectrum of reduced, intact enzyme complex. The demonstrates that the immediate environment of th cluster is little perturbed by dissociating this subunit from the FAD containing Mr 79,000 subunit. The temperature dependence of the power-saturation behaviour has, however, greatly decreased in the isolated subunit, the saturation at 11 K of the paramagnetic cluster being much less than in the enzyme complex. Moreover, the temperature dependence of th power-saturation behaviour of this cluster in the enzyme is greater with succinate as reducing agent, than with dithionite. (3) The [4Fe-4S] cluster is located on the Mr 79,000 subunit. This cluster is unstable in air when the subunit has been dissociated from the enzyme complex. PMID- 6271223 TI - Human erythrocyte cytosol phosphatidyl-inositol-bisphosphate phosphatase. AB - A phosphatidyl-myoinositol-4,5-bisphosphate phosphohydrolase (phosphatidyl inositol-bisphosphate phosphatase, EC 3.1.3.36) was detected in human erythrocytes and partially purified from the cytosol. Hemoglobin was removed by (NH4)2SO4 fractionation and chromatography on CM-Sepharose CL-6B. A 27,000-fold purification was achieved following gel filtration,, ion-exchange chromatography and hydrophobic chromatography. Although the preparation was not homogeneous, the molecular mass of the enzyme was estimated to be 105,000 by gel filtration. The activity was stabilized by a non-ionic detergent (Triton X-100). The enzyme was active with PI-P2 and, to a lesser extent, myo-inositol 1, 4, 5-trisphosphate but not with PI-P nor a variety of other lipid and non-lipid phosphate esters. In the presence of both cationic and non-ionic detergents, the effects of divalent cations were independent of substrate concentration. Mg2+ was required ('apparent' Km = 12 muM). The 'apparent' Km for the substrate was 0.27 mM and the specific activity was 765 +/- 191 (S.D.) nmol/min per mg protein. Inhibition by Ca2+ ('apparent' Ki = 50 microM) was competitive with Mg2+. Neomycin was an inhibitor at 10(-6) - 10(-4) M but only in the absence of Triton X-100. The phosphatase was inhibited by hemoglobin at concentrations higher that 1% (w/v) and by agents which react with sulfhydryl groups, but was unaffected by dithioerythritol and F-. PMID- 6271224 TI - New synthetic inhibitors of C1r, C1 esterase, thrombin, plasmin, kallikrein and trypsin. AB - p-Guanidinobenzoate derivates were prepared and their inhibitory effects on trypsin, plasmin, pancreatic kallikrein, plasma kallikrein, thrombin, C1r and C1 esterase were examined. Among the various inhibitors tested, 6'-amidino-2 naphthyl-4-guanidinobenzoate dihydrochloride, 4-(beta-amidinoethenyl)phenyl-4 guanidinobenzoate dimethanesulfonate and 4-amidino-2-benzoylphenyl-4 guanidinobenzoate dimethanesulfonate were the most effective inhibitors of trypsin, plasmin, pancreatic kallikrein. plasma kallikrein and thrombin and they strongly inhibited the esterolytic activities of C1r and C1 esterase, and then strongly inhibited complement-mediated hemolysis. PMID- 6271225 TI - Human fetal brain cells in culture. Increase in GM2 ganglioside after SV-40 transformation. PMID- 6271226 TI - Receptor-mediated gonadotropin action in the ovary. Demonstration of acute dependence of rat luteal cells on exogenously supplied steroid precursor (sterols) for gonadotropin-induced steroidogenesis. AB - Incubation of luteal cells with human, horse and rat sera, but not bovine sera resulted in enhanced basal and hCG-stimulated progesterone accumulation. The stimulatory effect of human or rat sera on basal, hCG- or 8 Br-cyclic AMP-induced progesterone synthesis in luteal cells was evident within 15-30 min after incubation, reaching a maximum after 3-4 h. The stimulatory effects of hCG and/or sera were blocked by inhibitors of RNA and protein synthesis. Similarly, lysosomotropic agents, chloroquine (100 microM) and ammonium chloride (10 mM), partly blocked the steroidogenic response of luteal cells to hCG and/or human or rat sera. Incubation of cells in the presence of 2-deoxyglucose, sodium azide and phenylmethylsulfonyl fluoride resulted in partial inhibition of progesterone secretion in response to hCG or sera. Fractionation of human or rat sera into various lipoprotein fractions demonstrated that LDL and HDL most effectively supported and potentiated the steroidogenic response to hCG. Lipoprotein deficient serum, however, did not alter gonadotropin-induced steroid production. Incubation of luteal cells with increasing concentrations of h-LDL and h-HDL enhanced both basal and hCG-mediated steroidogenesis in a dose-related manner, although very high concentrations of these lipoproteins were inhibitory. Further, [3H]cholesterol from [3H]cholesteryl linoleate-LDL was incorporated into luteal cell progesterone and the extent of this incorporation was enhanced by hCG. Addition of excess unlabeled h-LDL, h-HDL, as well as r-HDL, drastically reduced the incorporation of radioactive label into progesterone. These studies suggest that (a) serum potentiation of steroidogenesis was due to presence of lipoproteins, mainly LDL and HDL, and (b) the lipoprotein-bound cholesterol is delivered into the luteal cells and utilized for steroidogenesis. PMID- 6271227 TI - Uptake of lipophilic carcinogens by plasma lipoproteins. Structure-activity studies. AB - This report describes the interaction between plasma lipoproteins and two hydroxylated metabolites of benzo[a]pyrene, 3-hydroxybenzo[a]pyrene and benzo[a]pyrene-7,8-dihydrodiol, which differ significantly in lipophilicity. When incubated with plasma, the metabolites of benzo[a]pyrene exhibit a decreasing distribution into the ultracentrifugal lipoprotein fraction (d less than or equal to 1.20) and an increasing distribution into the albumin-rich fraction (d greater than 1.20) as the degree of hydroxylation of the metabolite increases. At saturation, uptake of benzo[a]pyrene by VLDL, LDL and HDL correlates with lipoprotein and total-lipid volume. Uptake of hydroxylated derivatives per lipoprotein total-lipid volume, in general, decreases with increasing hydroxylation. Contrary to this trend, HDL uptake of 3-hydroxybenzo[a]pyrene at saturation is significantly higher than its uptake of benzo[a]pyrene. Uptake of benzo[a]pyrene-7,8-dihydrodiol per total-lipid volume by all of the lipoprotein classes at saturation is considerably lower than their uptake of 3 hydroxybenzo[a]pyrene. Factors in addition to lipid solubility substantially alter lipoprotein uptake of the metabolites. PMID- 6271228 TI - Characterization of phosphatidylserine synthase from Saccharomyces cerevisiae and a mutant defective in the enzyme. AB - The membrane fraction of exponentially growing cells of Saccharomyces cerevisiae was found to exhibit phosphatidylserine synthase activity. The enzyme was solubilized by Triton X-100 and chromatographed on a Sepharose 6B column. The enzyme had a pH optimum between 8.0 and 8.5. The apparent Km values for CDPdiacylglycerol and L-serine were 0.12 and 13 mM, respectively. Triton X-100 stimulated the enzyme. Mg2+ or Mn2+ was required for the activity. Ca2+ was inhibitory at relatively low concentrations. The enzyme was highly specific to L serine. Labeling experiments showed that the enzyme synthesized phosphatidylserine by transferring the phosphatidyl moiety to L-serine. A mutant of S. cerevisiae defective in phosphatidylserine synthase was isolated. The strain required ethanolamine for its growth. Ethanolamine could be substituted by choline or high concentrations of L-serine. The mutant showed normal levels of CDPdiacylglycerol-inositol 3-phosphatidyltransferase and phosphatidylethanolamine methyltransferase activities. PMID- 6271229 TI - Effects of dibutyryl cyclic AMP and theophylline on rat pancreatic phospholipids in vitro. Ca2+-sensitive decrease in phosphatidylinositol and cycloheximide sensitive increase in phosphatidic acid. AB - We evaluated the effects of dibutyryl cyclic AMP and theophylline on rat pancreatic phospholipid metabolism in vitro. Dibutyryl cyclic AMP decreased mean phosphatidylinositol concentration by 30%, increased phosphatidic acid and phosphatidylglycerol concentrations by 90 and 25%, respectively, and increased [32P]phosphate incorporation into phosphatidic acid and phosphatidylinositol several-fold. Theophylline provoked similar changes in phosphatidic acid and phosphatidylinositol concentrations, and both stimulatory agents enhanced amylase and insulin secretion. Effects of dibutyryl cyclic AMP on amylase secretion and phospholipid levels were dependent on Ca2+. Cycloheximide blocked induced increases in phosphatidic acid, but did not diminish phosphatidylinositol breakdown or amylase secretion. Contrary to previous postulations, the present findings suggest: (a) cyclic AMP provokes large-scale phosphatidylinositol breakdown in the pancreas; (b) this phosphatidylinositol breakdown is dependent on Ca2+; and (c) phosphatidylinositol breakdown may contribute to exocytosis. In addition, it appears that a labile protein is required for synthesis of phosphatidic acid from 1,2-diacylglycerol and ATP. PMID- 6271230 TI - Diethylstilbestrol treatment modulates the enzymatic activities of phosphatidylcholine biosynthesis in rooster liver. AB - The effect of diethylstilbestrol injection on the activities of phosphatidylcholine biosynthetic enzymes in rooster liver has been determined. Choline kinase activity was stimulated within 4 h after the first hormone injection. By the third day enzyme activity reached 5.47 nmol . min-1 . mg-1 protein compared to control values (1.83 nmol . min-1 . mg-1 protein) which were unchanged during the the experiment. CTP : phosphocholine cytidylyltransferase activity was unaffected until Day 3 when its activity was 50% that of control values. When assayed in the presence of exogenous phospholipid, no significant change was noted in cytidylyltransferase activity. The activity of CDPcholine : 1,2-diacylglycerol phosphocholinetransferase was not altered by the hormone injections. The activity of phosphatidylethanolamine-N-methyltransferase gradually increased so that by Day 3, the enzyme activity was elevated 2-fold (0.12 to 0.24 nmol methyl group transferred per mg microsomal protein). These results are consistent with earlier in vivo studies (Vigo, C. and Vance, D.E. (1981) Eur. J. Biochem., in the press) that indicated a stimulation of phosphatidylcholine biosynthesis via CDPcholine during the first 2 days of diethylstilbestrol injection and inhibition on the third day. PMID- 6271231 TI - Topography of glycerolipid synthetic enzymes. Synthesis of phosphatidylserine, phosphatidylinositol and glycerolipid intermediates occurs on the cytoplasmic surface of rat liver microsomal vesicles. AB - The topography of glycerolipid biosynthetic enzymes within the transverse plane of rat liver microsomal vesicles was investigated: (1) by use of the impermeant inhibitor, mercury-dextran; (2) by use of proteases; and (3) by determining whether the enzyme activities are latent. The seven enzyme activities investigated (dihydroxyacetone-phosphate acyltransferase, acyldihydroxyacetone phosphate oxidoreductase, phosphatidic acid : CTPcytidyltransferase, CDPdiacylglycerol : inositol phosphatidyltransferase, 2-monoacylglycerol acyltransferase, diacylglycerol kinase, and the serine base exchange enzyme) function in phosphatidylinositol and phosphatidylserine synthesis and at intermediate levels in glycerolipid synthesis including steps of ether lipid synthesis. Mercury-dextran inhibited four of these enzymes greater than 60% in intact microsomal vesicles. One or more of the proteases employed (chymotrypsin, trypsin and pronase) inactivated each of the seven enzyme activities in intact microsomal vesicles. These two approaches indicate that each of these enzymes has important domains located on the cytoplasmic surface of microsomal vesicles. These enzyme activities could be assayed in intact microsomal vesicles. None appeared to be highly latent, indicating that substrates have free access to active sites. One substrate for each of these enzymes had been shown previously to be unable to cross the microsomal membrane. These data indicate that the active sites of these enzymes are located on the cytoplasmic surface of microsomal vesicles. It is concluded that the synthesis of phosphatidylserine and phosphatidylinositol, intermediates of ether lipid formation and other intermediates of glycerolipid synthesis occur asymmetrically on the cytoplasmic surface of the endoplasmic reticulum. These findings and our previous investigations on the topography of seven enzymes of triacylglycerol, phosphatidylcholine and phosphatidylethanolamine biosynthesis (Ballas, L.M. and Bell, R.M., Biochim. Biophys. Acta 602, (1980) 578-590) indicate that the synthesis of the major cellular glycerolipids occurs asymmetrically on the cytoplasmic surface of the endoplasmic reticulum. PMID- 6271232 TI - Uptake of chylomicron remnants by the native LDL receptor in human monocyte derived macrophages. AB - Human chylomicron remnants were taken up by cultured human monocyte-derived macrophages. Competition studies using 125I-labeled and unlabeled lipoproteins demonstrated that the remnant particles were not taken up by the modified LDL (acetyl LDL) receptor in these cells, which also contain a receptor for native LDL. The data thus suggest that the apolipoprotein E- and B-containing remnant particles are mainly taken up through an extra-hepatic E and B receptor (the classical LDL receptor pathway) in macrophages as is the case in cultured human skin fibroblasts. PMID- 6271233 TI - Catabolism of low-density lipoprotein by fibroblasts cultured in medium supplemented with saturated or unsaturated free fatty acids. AB - The increase in the degree of unsaturation of phospholipid fatty acids that occurred when fibroblasts were grown in medium supplemented with linoleic acid was associated with an increase in the degradation of LDL by both receptor mediated and receptor-independent pathways. There was not a corresponding increase in the number of surface receptors for LDL. PMID- 6271235 TI - Characterization of phospholipids and localization of some phospholipid synthetic and subcellular marker enzymes in subcellular fractions from rabbit lung. PMID- 6271234 TI - Hormonal mediation of rat heart lipoprotein lipase activity after fat feeding. AB - The effect of acute fat feeding on the response of two fractions of lipoprotein lipase in heart was explored. In rats, previously fasted, lipoprotein lipase activity released into the perfusate by heparin increased approximately 50% 4 h after fat feeding. The lipase activity remaining in the heart tissue after heparin perfusion showed no significant difference. When rats maintained ad libitum were intubated with glucose 2 h before the fat dose, a relatively larger increase (5-10-fold) in the heparin-releasable lipase activity was observed. The capacity of these hearts to hydrolyze 14C-labeled chylomicrons was also increased 4-5-fold over the controls. Fat ingestion has been reported to elevated plasma corticosteroid levels in rats. When adrenalectomized rats were fed fat, no significant changes in the heparin-releasable lipase activity were observed Hydrocortisone and corticotropin treatment increased the heparin-releasable lipase activity to the same degree as observed with fat feeding. These data suggest that the increase in heart lipoprotein lipase activity following fat feeding is mediated via corticosteroids. PMID- 6271237 TI - Biosynthesis of sphingomyelin from erythro-ceramides and phosphatidylcholine by a microsomal cholinephosphotransferase. AB - Mouse liver microsomes were shown to be active in the synthesis of sphingomyelin from ceramide and phosphatidylcholine in a reaction independent of CDPcholine. The conversion was not inhibited by calcium chelating reagents, and no evidence for the involvement of phospholipase C activity in the transformation could be adduced. Activity was also demonstrated in monkey liver and heart microsomes. Mouse brain microsomes produced a sphingomyelin analogue, tentatively identified as ceramide phosphorylethanolamine, but not sphingomyelin. Both [14C]ceramide and [G-14]phosphatidylethanolamine were precursors of the brain product, while phosphatidylcholine was inactive. Progress in the partial characterization of the liver enzyme is also described. PMID- 6271236 TI - Calcium-dependent turnover of brain polyphosphoinositides in vitro after prelabelling in vivo. AB - Rat brain phospholipids were labelled in vivo by an intraventricular injection of 32P. The radioactivity was found to accumulate predominantly in limbic structures, particularly hippocampus and diencephalon. A rapid and high specific labelling of the inositol phospholipids and phosphatidic acid was observed. The rate of incorporation into a crude myelin fraction was similar to that into a mitochondrial/synaptosomal fraction although phosphatidyl-myo-inositol 4,5 diphosphate was especially enriched in myelin. Upon incubation in vitro high specific labelling of the inositol phospholipids and phosphatidic acid was observed. The rate of incorporation into a crude myelin fraction was similar to that into a mitochondrial/synaptosomal fraction although phosphatidyl-myo inositol 4,5-diphosphate was especially enriched in myelin. Upon incubation in vitro high specific labelling of the inositol phospholipids and phosphatidic acid was observed. The rate of incorporation into a crude myelin fraction was similar to that into a mitochondrial/synaptosomal fraction although phosphatidyl myo-inositol 4,5-diphosphate was especially enriched in myelin. Upon incubation in vitro of the brain fraction after 2 h prelabelling in vivo, both phosphatidyl myo-inositol 4-phosphate and phosphatidyl-myo-inositol 4,5-diphosphate rapidly lost their radioactivity. Half of the labile fraction of the incorporated 32P was removed within 2 min. None of the other phospholipids changed in the 30 min in vitro incubation period. The metabolism of the polyphosphoinositide proceeded at a lower rate when the temperature was lowered, and was Ca2+-dependent. Further subcellular fractionation revealed that purified synaptosomes and myelin contained highly labelled phosphatidyl-myo-inositol 4-phosphate or phosphatidyl myo-inositol 4,5-diphosphate. Mitochondria contained highly labelled phosphatidyl myo-inositol but no phosphatidyl-myo-inositol 4-phosphate or phosphatidyl-myo inositol 4,5-diphosphate. ACTH1-24 did not inhibit the in vitro dephosphorylation of prelabelled polyphosphoinositide, confirming previous findings that the peptide affects the polyphosphoinositide kinases and not the respective phosphatases. PMID- 6271238 TI - Kinetic and equilibrium studies of alkaline isomerization of vertebrate cytochromes c. AB - Equilibria and kinetics of alkaline isomerization of seven ferricytochromes c from vertebrates were studied by pH-titration and pH-jump methods in the pH region of 7-12. In the equilibrium behavior, no significant difference was detected among the cytochromes c, whereas marked differences in the kinetic behavior were observed. According to the kinetic behavior of the isomerization, the cytochromes c examined fall into three classes: Group I (horse, sheep, dog and pigeon cytochromes c), Group II (tuna and bonito cytochromes c) and Group III (rhesus monkey cytochrome c). The kinetic results are interpreted in terms of the sequential scheme: Neutral form in equilibrium with fast Transient form in equilibrium with slow Alkaline form where the neutral and alkaline forms are the species stable at neutral and alkaline pH, respectively, and the transient form is a kinetic intermediate. From comparison of the primary sequences of the seven cytochromes c and the classification of these cytochromes c, it is concluded that the amino acid substitution Phe/Tyr at the 46-th position has a major influence on the kinetic behavior. In Group II and III cytochromes c, the ionization of Tyr 46 is suggested to bring about loosening of the heme crevice and thus facilitate the ligand replacement involved in the isomerization. PMID- 6271239 TI - Electron paramagnetic resonance studies of Pseudomonas cytochrome c peroxidase. AB - The EPR spectrum at 15 K of Pseudomonas cytochrome c peroxidase, which contains two hemes per molecule, is in the totally ferric form characteristic of low-spin heme giving two sets of g-values with gz 3.26 and 2.94. These values indicate an imidazole-nitrogen : heme-iron : methionine-sulfur and an imidazole-nitrogen : heme-iron : imidazole-nitrogen hemochrome structure, respectively. The spectrum is essentially identical at pH 6.0 and 4.6 and shows only a very small amount of high-spin heme iron (g 5--6) also at 77 K. Interaction between the two hemes is shown to exist by experiments in which one heme is reduced. This induces a change of the EPR signal of the other (to gz 2.83, gy 2.35 and gx 1.54), indicative of the removal of a histidine proton from that heme, which is axially coordinated to two histidine residues. If hydrogen peroxide is added to the partially reduced protein, its EPR signal is replaced by still other signals (gz 3.5 and 3.15). Only a very small free radical peak could be observed consistent with earlier mechanistic proposals. Contrary to the EPR spectra recorded at low temperature, the optical absorption spectra of both totally oxidized and partially reduced enzyme reveal the presence of high-spin heme at room temperature. It seems that a transition of one of the heme c moieties from an essentially high-spin to a low spin form takes place on cooling the enzyme from 298 to 15 K. PMID- 6271240 TI - Amino acid sequence determination of the ADP,ATP carrier from beef heart mitochondria. The sequence of the C-terminal acidolytic fragment. AB - The primary structure of the C-terminal region (94 residues) of the ADP,ATP carrier of beef heart mitochondria is described. CNBr cleavage results in a large peptide (CB1) with Mr 22 000 and several small peptides (CB2 to CB8). Peptide separation was achieved by gel chromatography with 80% formic acid or with an ethanol/formic acid mixture. The amino acid sequence of the small CNBr peptides was determined by solid-phase techniques. Hydrolysis in formic acid cleaves the carrier protein into an Mr 23 000 fragment (A1) with the blocked N-terminus and an Mr 10 000 fragment (A2) starting with proline. The alignment of two CNBr fragments was possible by degradation of A2 by solid-phase methods for 34 steps. The remaining CNBr fragments were arranged by sequencing the tryptic peptides of citraconylated A2. PMID- 6271241 TI - Fluorescent labeling of the carbohydrate moieties of human chorionic gonadotropin and alpha 1-acid glycoprotein. AB - A method for covalent attachment of a fluorescent molecule to the carbohydrate moieties of glycoproteins is described. The glycoproteins were oxidized with periodate under mild conditions selective for sialic acid (Van Lenten, L. and Ashwell, G. (1971) J. Biol. Chem. 246, 1889--1894). The resulting aldehydes were condensed with either dansylhydrazine, dansylethylenediamine, or fluoresceinamine followed by reduction with NaCNBH3 and NaBH4. Conjugates prepared with dansylhydrazine were found to be insufficiently stable for spectroscopic analysis, whereas the primary amines produced stable conjugates whose fluorescence polarization (P) was constant for several hours at 37 degrees C. The degree of labeling correlated roughly with the sialic acid contents of the vaious glycoproteins. Very little covalent incorporation was observed with albumin (which is devoid of carbohydrate) or with asialo alpha 1-acid glycoprotein. Exclusion chromatography in the presence of a dissociating agent was sometimes required to remove significant amounts of noncovalently adsorbed dye. Fluorescent labeled alpha subunits of human chorionic gonadotropin were shown to recombine normally with native beta subunits. However, the labeling procedure appeared to compromise the ability of the beta subunits to recombine. Electrophoretic analysis produced evidence of covalent cross-linking between subunits following periodate oxidation of the intact gonadotropin. The possibility that primary amine groups of the protein compete with added fluorescent amines for reaction with periodate-generated aldehydes is discussed. PMID- 6271242 TI - Application of high--performance liquid chromatography to abnormal hemoglobin studies. Characterization of hemoglobins D in Ivory Coast and description of a new variant hb Cocody (beta 21 (B3) Asp leads to Asn). PMID- 6271243 TI - Cell adhesion-dependent differences in endogenous protein phosphorylation on the surface of various cell lines. AB - Endogenous phosphorylation of intact cells was studied with four mouse, hamster and human cell lines using [gamma-32P]ATP and [gamma-32P]GTP as exogenous substrates. With all four cell lines distinct differences in the phosphoprotein patterns could be demonstrated for cells grown in suspension culture compared to cells grown in monolayers. Two major, apparently ubiquitous phosphoproteins with molecular weights of 135 000 (128 000 in HeLa cells) and 105 000, representing up to 60% of total phosphorylation, were phosphorylated only in cells grown in suspension. These phosphoproteins and the kinase(s) were located on the surface of the suspension cells. Evidence showed that phosphorylation was apparently not a true endogenous reaction, that rather it occurred by cell-cell collision, showing exponentially increasing 32P incorporation with increasing cell population density. Phosphorylation of pp135 and pp105 was established with ATP as well as with GTP and was not dependent on cyclic nucleotides cyclic AMP, cyclic GMP and cyclic CMP. The substrate-attached cells of all four cell lines have protein kinases on the cell surface. The lack of pp135 and pp105 phosphorylation may be due to the fact that these phosphoproteins are not expressed at all on the surface of substrate-attached cells or that these phosphoproteins are already fully phosphorylated. PMID- 6271244 TI - Evidence for protein self-association induced by excluded volume. Myoglobin in the presence of globular proteins. AB - The fluorescence polarization of fluorescent derivatives of hemoglobin and myoglobin was measured as a function of the concentration of added polymers (PEG 6 000, PEG-20 000) and globular proteins (lysozyme, ribonuclease A, beta lactoglobulin). The results indicated that the effective size and shape of 1 anilino-9-naphthalene sulfonate myoglobin are unaltered in the presence of up to 25 g/dl poly(ethylene glycol), whereas they are significantly altered in the presence of comparable concentrations of other proteins. The results are consistent with the hypothesis that in the presence of high concentrations of added protein, 1-anilino-9-naphthalene sulfonate myoglobin self-associates to form a dimer similar in size and shape to 1-anilino-9-naphthalene sulfonate hemoglobin. PMID- 6271245 TI - Temperature-dependent and transformation-associated changes in polyamine metabolism in normal and Rous sarcoma virus-infected chick embryo fibroblasts. AB - Tertiary cultures of chick embryo fibroblasts infected and transformed by the wild-type Rous sarcoma virus, when actively growing at 35 degrees C, had higher putrescine levels than the respective uninfected cells. Transformed cells also had much higher specific activity of ornithine decarboxylase (EC 4.1.1.17) than the normal fibroblasts. At 41 degrees C the difference in putrescine levels between the normal and the transformed cells was less marked, and both cell types showed a relative accumulation of spermine. Cultures infected with the NY68 mutant virus, which is temperature-sensitive for transformation, showed at 41 degrees C normal cell morphology and intermediate polyamine patterns, while at 35 degrees C a transformed phenotype was found in both aspects. In shift-down experiments a change towards the permissive temperature pattern of polyamine metabolism was evident within 2-3 h. Difluoromethylornithine, a specific and irreversible inhibitor of ornithine decarboxylase efficiently reduced the enzyme activity as well as the levels of both putrescine and spermidine in all culture types and temperatures. Incubation of Rous sarcoma virus-transformed cells with 3 mM difluoromethylornithine for 36 h did not affect the maintenance of the transformed state. Likewise, when NY68-infected cultures were exposed to difluoromethylornithine at 41 degrees C for 12 h and then shifted down to 35 degrees C, the appearance of the transformed morphology took place concomitantly with that of the control cultures without respective changes in the polyamine levels. This suggests that the transformation-associated pattern of polyamines in chick embryo fibroblasts is not a prerequisite for morphological transformation of these cells. PMID- 6271246 TI - Regulation by human chorionic gonadotropin of protein kinases in rabbit endometrium. AB - The effect of human chorionic gonadotropin (CG) on protein kinase levels has been studied in rabbit endometrium. A good correlation was observed between alterations of total protein kinase activity and DNA content in the tissue. The level of type I cyclic AMP-dependent protein kinase had slightly increased by 6 h after human CG treatment, and then decreased later. In contrast, the type II enzyme gradually increased after 3 days. The increase of the activity at 7 days was mainly due to that of type II enzyme, and the ratio of type II to type I enzyme increased. The activity pattern at the later stage closely resembled that of treatment with progesterone. PMID- 6271247 TI - Glucocorticoids and aspirin inhibit and cyproterone acetate enhances the androgenic response in mouse kidney. AB - Glucocorticoids and aspirin antagonize the androgenic response in mouse kidney, but not in ventral prostate or seminal vesicles. These agents impeded the testosterone-mediated increase in kidney weight, cytochrome c oxidase, and lysosomal hydrolases and urinary excretion of lysosomal hydrolases and proteins. They also attenuated the testosterone-induced decrease in enzyme latency and membrane stability of kidney lysosomes. In contrast, the antiandrogen cyproterone acetate is weakly androgenic in kidney and potentiates testosterone-induced lysosomal enzymuria and proteinuria (synandrogenic effect). PMID- 6271248 TI - Evidence for two types of beta-adrenergic-sensitive adenylate cyclase activities in bovine cerebellum. AB - A latent, as well as an expressed form of adenylate cyclase coupled to beta adrenergic receptors is present in intact crude synaptosomal preparations from bovine cerebellum. The latent adenylate cyclase activity was assayed in Krebs Ringer buffer by [3H]adenine labeling and was found to be coupled to a beta 1 like adrenergic receptor. The externally accessible adenylate cyclase assayed in the same medium with [3H]ATP was stimulated via beta 2-adrenergic receptors. PMID- 6271249 TI - Theophylline reduces poly (ADP-ribose) synthetase from chick embryo liver nuclei. AB - The effect of theophylline on poly(ADP-ribosyl)ation was investigated. The poly(ADP-ribose) synthetase activity in vitro was markedly reduced in the liver nuclei prepared from theophylline-treated chick embryo. This reduction was not due to the enzyme inhibition by theophylline contamination in the nuclear fraction. The hydroxyapatite column chromatographic analysis of [3H]adenosine labelled poly(ADP-ribose)molecules formed in vivo revealed that the in vivo formation of poly(ADP-ribose)molecules was also decreased by theophylline administration. The theophylline-induced reduction of poly(ADP-ribose) synthesis was not due to either low NAD levels or to a decrease in the chain length of the poly(ADP-ribose) molecule, rather this reduction was derived from a decrease in the number of poly(ADP-ribose) molecule. Possible mechanisms related to reduction of poly(ADP-ribose) synthesis in vivo are discussed. PMID- 6271251 TI - Levels of collagenolytic activity, beta-glucuronidase, and collagen prolyl hydroxylase in paws from rats with developing adjuvant arthritis. AB - The collagenolytic activity associated with insoluble collagen fibers separated from homogenates of inflamed paws from rats with adjuvant arthritis was quantitated using EDTA-sensitive solubilization of hydroxyproline as a measure of activity. Approximately 60% of the solubilized hydroxyproline was associated with dialyzable products. The level of collagenolytic activity in the paws increased with time after the induction of adjuvant arthritis and paralleled to a large extent the development of inflammation in both the adjuvant injected (right) hind paw and in the non-injected, contralateral paw. By day 26, the level of free collagenolytic activity in the injected paw had increased to a level 30 times normal while that in the contralateral paw had increased to a level 10 times normal. Treatment of the residues from the injected paws with trypsin resulted in the activation of a latent collagenolytic activity which, on day 26, accounted for approximately 50% of the total activity. The elevated level of collagen prolyl hydroxylase in the inflamed paw suggested that the rate of collagen synthesis was also increased. The activity of beta-glucuronidase increased in the inflamed paw with time after the induction of adjuvant arthritis while that of cathepsin G was elevated as compared to normal in paws removed, 5 but not 22 days after the induction of adjuvant arthritis. The inflamed paw of the adjuvant rat may represent a useful system in which to study the role of collagenolytic enzymes in the destruction of connective tissue by inflammatory lesions. PMID- 6271250 TI - Dynamics of calmodulin and cyclic AMP phosphodiesterase in plasma membranes of rat livers and ascites hepatomas. AB - 1. Plasma membranes from ascites hepatoma cells (AH-7974, AH-130) contained much smaller amounts of calmodulin (about half) and cyclic AMP phosphodiesterase (about one-third) compared to plasma membranes of rat livers. 2. Some of calmodulin molecules in liver plasma membranes were released by repeated washing. The 'washed' liver plasma membranes showed the presence of specific binding sites for externally added calmodulin molecules (bovine brain) (N = 140 pmol/mg protein, Kd = 7.9 . 10(-8) M). The calmodulin content of AH-7974 plasma membranes was not reduced by repeated washing. The binding of calmodulin to the 'washed' AH 7974 plasma membranes was only of nonspecific nature with negative cooperativity. 3. Plasma membranes (liver and AH-7974) appeared to contain both calmodulin dependent and calmodulin-independent phosphodiesterase, but the stimulation by externally added Ca2+ plus calmodulin was rather small. Externally added calmodulin-dependent phosphodiesterase (bovine brain) was bound more to 'washed' liver plasma membranes than to 'washed' AH-7974 plasma membranes. Newly bound phosphodiesterase appeared to be more sensitive to the stimulation by Ca2+ plus calmodulin in 'washed' hepatoma plasma membranes than in 'washed' liver plasma membranes. 4. Preincubation of 'washed' plasma membranes (liver and hepatoma) with calmodulin did not affect the binding of phosphodiesterase, but the sensitivity of phosphodiesterase to the stimulation by Ca2+ plus calmodulin in hepatoma plasma membranes was lost. PMID- 6271252 TI - Effects of melittin on adipocyte metabolism unrelated to lysophospholipid accumulation. AB - Melittin addition to rat or hamster adipocytes resulted in inhibition of lipolysis, cyclic AMP accumulation and glucose oxidation. Low concentrations of melittin were not insulin-like with respect to either stimulation of glucose metabolism or inhibition of lipolysis. Higher concentrations of melittin lysed adipocytes. In the presence of melittin, cellular phospholipids were released to the medium and hydrolyzed with little accumulation of lysophospholipids. Only in adipocytes incubated with melittin contaminated with phospholipase A2 was any appreciable accumulation of lysophospholipids seen and this was in the medium. These data suggest that the toxic effects of melittin on adipocytes are not due to the accumulation of lysophospholipids but rather to the loss of membrane phospholipids or alterations in membrane proteins. PMID- 6271253 TI - Extracellular folate deaminase of Dictyostelium discoideum. AB - Folate deaminase released from cells of Dictyostelium discoideum is heterogeneous with respect to molecular weight and stability at 60 degrees C. The most heat stable component isoelectrofocuses in a broad band at approx. pH 6. The Km value of this component for folate is approx. 7 x 10(-7)M and Mr approx. 40 000. The major portion if not all of the deaminase binds to immobilized concanavalin A and lentil lectin. Extracellular folate deaminase has a pH-optimum of approx. pH 6.0. This is higher than that of lysosomal enzymes, which are also glycoproteins released into the extracellular medium. PMID- 6271254 TI - Role of sulfhydryl oxidation in adipocyte plasma membrane surface in the response of adenylate cyclase to isoproterenol and glucagon. PMID- 6271255 TI - Iron transfer form transferrin to ferritin mediated by polyphosphate compounds. AB - We have studied iron transfer from transferrin to ferritin in the presence of ATP, GTP, ADP, AMP and 2,3-diphosphoglycerate. These compounds, with the exception of AMP, can release iron from transferrin at pH 7.4 and form a stable Fe(III)-phosphate complex. From these complexes, only a limited number of Fe(III) atoms can be incorporated into ferritin. Ascorbate enhances iron transfer from transferrin to ferritin at the beginning of the process but subsequently inhibits further iron deposition in ferritin. PMID- 6271256 TI - Modulation by retinoic acid and corticosteroids of collagenase production by rabbit synovial fibroblasts treated with phorbol myristate acetate or poly(ethylene glycol). PMID- 6271257 TI - The roles of extracellular and intracellular calcium in lysosomal enzyme release and superoxide anion generation by human neutrophils. PMID- 6271258 TI - [Interrelationship of polyphosphate metabolism and levorin biosynthesis in Streptomyces levoris]. AB - The effect of inorganic phosphate on biosynthesis of the polyene antibiotic levorin by Streptomyces levoris was studied. At phosphate concentration of 4.0 mM levorin biosynthesis is repressed by 90%, resulting in an increase of ATP and a condensed inorganic polyphosphates content in the producer cells. At phosphate concentration optimal for levorin production (0.04 mM) the level of intracellular ATP sharply falls by the beginning of the steady-state phase of the producer growth and that of polyphosphates decreases 3-6-fold. The inorganic phosphate exerts different effects on polyphosphate metabolism enzymes, such as polyphosphate: D-glucose-6-phosphotransferase, polyphosphate phosphohydrolase, tripolyphosphate phosphohydrolase, pyrophosphate phosphohydrolase, alkaline and acid phosphatase. The strongest effect of phosphate excess is observed in the case of polyphosphate: D-glucose-6-phosphotransferase, whose activity decreases 2 5-fold. The activity of this enzyme was shown to be correlated with the antibiotic accumulation. The data obtained are indicative of interrelationship between the polyphosphate metabolism and levorin biosynthesis. PMID- 6271259 TI - [Interaction between free fatty acids and Ca2+-dependent ATPase from sarcoplasmic reticulum membranes]. AB - The interaction between free fatty acids and Ca2+-dependent ATPase, an intrinsic protein of sarcoplasmic reticulum membranes, was studied with relevance to the changes in membrane permeability induced by free fatty acids. It was found that only unsaturated fatty acids increase the permeability of reticulum membranes for Ca2+, this effect being completely reversible. The increase in the membrane permeability by fatty acids is coupled to a generation of a channel for Ca2+ efflux under effect of Ca2+-dependent ATPase. The interaction between fatty acids and Ca2+-dependent ATPase was demonstrated by the protein fluorescence and electron paramagnetic resonance methods, using spin-labelled fatty acid derivatives. A model demonstrating the increase of sarcoplasmic reticulum membrane permeability for Ca2+ in the presence of the fatty acid-Ca2+-dependent ATPase complex is proposed. PMID- 6271260 TI - [Substrate specificity of Ca2+,Mg2+-dependent DNAase from sea urchin (Strongylocentrotus intermedius) embryos]. AB - Ca2+,Mg2+-dependent DNAse from sea urchin embryos is specific to the secondary structure of substrates irrespective of the nature of activating cations. The enzyme does not split synthetic single-stranded oligo and polynucleotides, such as d(pTpTpTpCpC), d(pGpGpTpTpT). d(pApApTpTpC), d(pGpApApTpTpC), d(pA)5-poly(dT), d(pApApTpTpC)-poly(dT), poly(dA) and poly (dT) and hydrolyses the double-stranded substrates poly d(AT), poly (dA) . poly (dT) and highly polymerized DNA. Native double-stranded DNA from salmon and phage T7 is split by the enzyme at a higher rate than that of denaturated DNA of salmon and single-stranded DNA of phage M13. The high rate of poly(dA) . poly(dT) and poly d(AT) hydrolysis and the stability of poly(dG) . poly(dC) to the effect of the enzyme suggest a certain specificity of the enzyme to the nature of nitrogenous bases at the hydrolyzed phosphodiester bond of the substrate. PMID- 6271261 TI - [Protein modulator of cyclic nucleotide-dependent protein kinases from the shrimp Palaemon adspersus]. AB - A thermostable protein modulator of cyclic nucleotide-dependent protein kinases was isolated and purified from the tissue of the shrimp Palaemon adspersus. The molecular weight, N-terminal amino acid and amino acid composition of the protein modulator were determined and its effects on cAMP-dependent protein kinase depending on substrate and cation concentrations were investigated. The protein modulator under study is a low molecular weight (15 000) acid protein, pI-435, which exerts both strong activating and inhibitory effects on cAMP-dependent protein kinase in the reactions of phosphorylation of protamine and histones, respectively. PMID- 6271262 TI - [Determination of molar content of creatine kinase in heart mitochondria by SH reagents]. AB - The maximal content of mitochondrial isoenzyme of creatine kinase (CK) in rat heart mitochondria does not exceed 12.5 moles per mole of ATP-ADP translocase. This value was obtained by titration of mitochondrial CK activity in aged mitochondria by 5,5'-dithiobis-(2-nitrobenzoate) (DTNB) and 2,4 dinitrofluorobenzene (DNFB) and by a more complex and accurate method. The essential thiol groups of membrane-bound mitochondrial CK (and its enzymic activity) can be specifically protected by phosphocreatine (12 mM) + ADP (1-5 mM) against inactivation by DTNB. Mitochondria with protected SH-groups of CK and with groups inactivated by DTNB were repeatedly incubated with DTNB under identical conditions and the number of additionally reacted sulfhydryl groups and the changes in CK activity were measured. The differences in the number of additionally reacted SH-groups correlated with the changes in the CK activity, which made it possible to calculate the molar ratios of mitochondrial CK to cytochrome c oxidase and ATP-ADP translocase (2.16 +/- (0.4): 1:2, respectively). PMID- 6271263 TI - [Biosynthesis of early enzymes induced by bacteriophage T4]. AB - The biosynthesis of early enzymes of DNA-ligase, RNA-ligase, DNA-polymerase, polynucleotide kinase, exonuclease A induced by bacteriophage T4amN82 was studied. The maximal activity of DNA-ligase was observed at the 60th min after the infection, while that of the other enzymes was revealed at the 90th min and reached 4, 45, 529, 120 and 78 units per mg of protein for DNA-ligase, RNA ligase, polynucleotide kinase, DNA-polymerase and exonuclease A, respectively. Bacteriophage T4amN82 induced the maximal biosynthesis of the tested enzymes, when E. coli B-23 cells were grown in medium I containing trypton bacto ("Merck", West Germany) and a yeast extract ("Difco", USA). Similar events were observed when E. coli B-23 cells infected with phage T4amN82 were grown in a medium (II) with casein hydrolysate and yeast extract. Ultrasonication used for the disruption of E. coli B-23 cells infected with bacteriophage T4 had no effect on the enzyme activities. PMID- 6271264 TI - [Circular dichroism of cAMP-dependent protein kinase from pig brain]. AB - The circular dichroism spectra of phosphorylated and non-phosphorylated forms of cAMP-dependent protein kinase from pig brain and those of the catalytical and regulatory subunits of the enzyme were studied. The percentage of the secondary structure components of the subunits was calculated. cAMP was shown to cause conformational changes of the enzyme. The conformation of the cyclic nucleotide within the cAMP--regulatory subunit complex was established. It was assumed that the conformation of the cAMP molecule during enzyme activation is subjected to inversion. PMID- 6271265 TI - [Effect of nitrite on cytochrome oxidase]. AB - Nitrite causes changes in the optical and EPR spectra of cytochrome oxidase from heart and alters the spectral, redox and basic properties of cytochrome c. No utilization of nitrite by cytochrome oxidase was observed. However, nitrite inhibits the superoxide dismutase and oxidase activities of the enzyme. Changes in the properties of cytochrome oxidase were observed under effect of some products of nitrite reduction, e. g. nitric oxide, hydroxylamine, hydrazine; nitrate has no effect on the optical and EPR spectra or on the enzyme activity. PMID- 6271266 TI - [Serratia marcescens endonuclease. Properties of the enzyme]. AB - Some physico-chemical properties of endonuclease (EC 3.1.4.9) from Serratia marcescens were studied and the amino acid composition of the enzyme was determined. The protein molecule was shown to contain one SH-group and one S-S bond, which renders it different from the well studied nuclease (EC 3.1.4.7) from Staph. pyogenes. The conditions for reconstitution of the S-S-bond by dithioerythritol for quantitative estimation of cysteine residues of the endonuclease molecule were selected. The N-terminal amino acid was found to be threonine. The UV spectra for the enzyme are typical for proteins; A 0,1% 1cm,280nm is 1.46, epsilon 25 degrees 280nm,pH7,4 is 47292 M-1 cm-1. The sedimentation coefficient in phosphate buffer sW, 20 degrees is 3.4 S, pI is 6.5 and 7.5. PMID- 6271267 TI - [Some properties of the reaction catalyzed by protein kinase bound to cardiac sarcolemma]. AB - A highly purified sarcolemmal fraction from rat heart consisted of closed inside out oriented vesicles and possessed high activities of Na+, K+-ATPase, adenylate cyclase and creatine kinase. Contaminations of sarcolemmal preparation by other membranous fractions were practically absent. This sarcolemmal fraction contained protein kinase tightly bound to the membrane. Substrates of the phosphorylation reaction catalyzed by this protein kinase were either endogenous sarcolemmal protein (proteins) with molecular weight of 11500 or exogenous protein--histone, type II. Phosphorylation of the endogenous but not of the exogenous substrate was completely independent of cyclic AMP. A kinetic analysis of the sarcolemmal protein kinase reaction with Mg[gamma-32P]ATP and histone as substrates revealed that the kinetic mechanism of this reaction is characterized by the following kinetic parameters: Km (Mg-ATP) = 12.1 microM; Km (histone) = 0.47 mg/ml; Ki (Mg ADP) = 15.6 microM. A comparison of experimental results to literary data allows to suggest that the sarcolemmal enzyme is virtually soluble protein kinase tightly bound to the sarcolemma. PMID- 6271268 TI - [Interaction of the non-histone protein PS1 with some chromatin components]. AB - The binding of non-histone protein from mouse spleen chromatin located in the sites highly sensitive to micrococcal nuclease and DNA-ase I, to DNA and histones was studied. The binding of the DNA-protein complexes to nitrocellulose filters demonstrated the absence of protein binding to DNA. A highly selective binding of protein PS1 to histones H1 and H2A and to one of the non-histone proteins (presumably HMG 14) was revealed. It is concluded that protein PS1 is incorporated into chromatin by the protein-protein interactions. PMID- 6271269 TI - [Role of cytostatic agents in the pathogenesis of neuro- and myopathies]. PMID- 6271270 TI - [Nonspecific phosphodiesterases of human leukocytes, blood platelets and serum]. AB - Phosphodiesterases from blood cells and serum can be subdivided in several groups according to substrate specificity, optimum pH and effects of inhibitors: 1) Acidic phosphodiesterase activities were not inhibited by EDTA, represented the whole p3'T hydrolysing activity, but only a part of the activity hydrolysing the other substrates (p5'T was not hydrolysed at acidic pH). This acid phosphodiesterase activity was high in white blood cells and platelets but very low in serum. 2) Neutral phosphodiesterase activity was prevalent in leucocytes when BpP and BMP were used as substrates. 3) Alkaline phosphodiesterase activity was characterized by substrate specificity at optimum pH and distribution in cells and serum: in serum there are phosphodiesterases hydrolysing all checked substrates (p3'T excepted) at optimum pH 9.0, whereas in blood cells alkaline phosphodiesterase activities are very low for all substrates (excepted for p Phi Pn and p5'T). In each cell and serum we have determined, for all phosphodiesterase activities, the linearity of activity of versus time and versus protein concentration, the effect of substrate and effector concentration and the heat stability. PMID- 6271271 TI - Gating current and potassium channels in the giant axon of the squid. AB - Gating current (Ig) underlying Na-channel activation is large enough to enable resolution of components both preceding and paralleling Na conductance (gNa) turn on. For large depolarizations (beyond +20 mV), an additional "slow phase" of Ig is observed during a time when Na activation is already complete, but when K channel opening is just becoming detectable. If Na- and K-channel gating are similar, the slow kinetics and long delay for K activation predict that K channel Ig must be relatively small and slow. Externally applied dibucaine almost totally blocks gNa and greatly reduces the fast (Na channel) Ig without altering gK or the Ig slow phase. The slow phase of Ig depends in part of the presence of functional K channels. Selective diminution in amplitude of the slow phase is consistently observed after a 30-min perfusion with both external and internal K free media, a procedure which destroys nearly all K channels. This decrease of Ig amounts to approximately 10% of the total charge movements at +40 to +80 mV, with gating charge and K channels disappearing in a ratio of less than 1 e- per picosiemens of gK. These findings are consistent with the idea that part of the Ig slow phase represents gating current generated by the early steps in K-channel activation. PMID- 6271272 TI - Isolation by restriction endonuclease digestion and base-specific affinity chromatography of rat-embryo DNA sequences disproportionately enriched in virogenic bromodeoxyuridine. AB - Control and bromodeoxyuridine-containing rat-embryo-cell DNA were digested by the restriction endonucleases Hpa II and Msp I and were subsequently analyzed by agarose-gel electrophoresis as well as DNA-affinity chromatography. By the former technique, it appeared that no substantial differences existed between the two DNA samples with respect to the amount or distribution of methylcytosine. On the other hand, it was obvious following base-specific DNA chromatography that the virogenic analog was markedly concentrated in particular nucleotide sequences which demonstrated a proportionately greater affinity for the (A-T)-specific adsorbent irrespective of digestion by either restriction endonuclease. PMID- 6271273 TI - "Enterolglucagon': a specific effect on gastric glands isolated from the rat fundus. Evidence for an "oxyntomodulin' action. AB - A newly isolated porcine glucagon-like biologically active intestinal peptide ("enteroglucagon') has been tested for its ability to stimulate the cyclic AMP system present in different tissue preparations from the rat: membranes prepared from liver or from isolated fundic glands as well as intact glands isolated from either the fundus or the antrum. Enteroglucagon (EG) was about 10 times less potent than pancreatic glucagon (G) in liver membranes, whereas it was about 20 times more potent than G in fundic membranes as well as in fundic glands, where it acts at doses as low as 3 X 10(-10) M. EG and G were practically ineffective in antral glands. It is concluded that EG is an "under-glucagon' in rat liver, whereas it is a "super-glucagon' in rat fundic glands. Accordingly, we propose to call this peptide "oxyntomodulin'. PMID- 6271275 TI - The use of the restriction endonuclease DdeI for mapping relataed DNAs. AB - Restriction endonuclease mapping of BK virus strain GS using DdeI has allowed us to correct and extend the previously published map. PMID- 6271274 TI - Molecular events in leukocyte chemotaxis: their possible roles in processing the chemical signal. PMID- 6271276 TI - Analysis of a restriction endonuclease map for bovine papillomavirus type 2 DNA. AB - A physical map was constructed for bovine papillomavirus type 2 DNA by the use of restriction endonucleases. A comparison between the genomes of bovine papillomavirus types 1 and 2 in regard to location and number of cleavage sites of seven enzymes is also presented. This comparison revealed similarities between the two genomes. PMID- 6271277 TI - A severe and a mild potato spindle tuber viroid isolate differ in three nucleotide exchanges only. AB - Fingerprint analyses of two potato spindle tuber viroid (PSTV) isolates causing severe and mild symptoms, respectively, in tomato exhibited defined differences in the RNase T1 and RNase A fingerprints. The complete sequencing of the mild isolate and the comparison of its primary structure with the previously established one of the pathogenic type strain revealed that oligonucleotides CAAAAAAG, CUUUUUCUCUAUCUUACUUG, and AAAAAAGGAC in the 'severe' strain are replaced by CAAUAAG, CUUUUUCUCUAUCUUUCUUUG, AAU, and AAGGAC in the 'mild' strain. Thus, three nucleotide exchanges at different sites of the molecule may change a pathogenic viroid to a practically non-pathogenic isolate. The possible correlation between the secondary structure in a defined region of the PSTV molecule and its pathogenicity for tomato is discussed. PMID- 6271278 TI - A method for sequencing single-stranded cloned DNA in both directions. AB - A DNA sequencing method has been developed whereby DNA that has been cloned in a single-stranded bacteriophage vector can be sequenced from both ends. The method involves first making a minus-strand sense template from a single-stranded insert in the vector M13mp2 using a flanking primer, and then sequencing the synthesized template using the dideoxynucleotide termination method (Sanger et al., 1977, 1980) with a second primer. Special conditions are described under which the first primer is easily removed after making the template, and sequencing in the opposite direction can be done in the normal way (Sanger et al., 1980) without separating the double strands. This method renders it possible to read up to twice the amount of sequence data from a long insert and also to check short inserts by producing complementary sequence patterns. PMID- 6271279 TI - Dissections and reconstructions of genes and chromosomes. Nobel lecture, 8 December 1980. PMID- 6271280 TI - Investigations on myelination in vitro: thyroid hormone receptors in cultures of cells dissociated from embryonic mouse brain. AB - Brain cells dissociated from 15-day-old embryonic mice and grown in culture contain both cytosolic and nuclear receptors for L-3,5,3'-triiodothyronine (L T3). KD values for L-T3 of 3.05 X 10(-9) M and 4.2 X 10(-9) M were determined with the cytosolic and nuclear receptors respectively. These cultured cells, which are suitable for studying the regulation of myelination by T3 in vitro, display a high specificity for L-T3 in that the receptors for L-T3 do not bind D T3, D-thyroxine, L-diiodothyronine, or DL-thyronine, and bind only small amounts of L-thyroxine. PMID- 6271281 TI - Alignment of cloned amiE gene of Pseudomonas aeruginosa with the N-terminal sequence of amidase. AB - A restriction enzyme map was constructed for 5.1-kb fragment of Pseudomonas aeruginosa DNA inserted into plasmid pBR322. Restriction enzyme sites were matched to the N-terminal amino acid sequence of amidase to obtain alignment of the amiE gene within the cloned fragment. PMID- 6271282 TI - Characterization of Alu family repetitive sequences which flank human beta-type globin genes. AB - Heteroduplex mapping has determined the size, location, and orientation of three Alu family sequences from the human beta-type globin gene cluster. Two of these sequences have the same orientation. One (231 bp long) is 2 kb to the 5' side of the B gamma-globin gene and the other (222 bp) is l kb 5' to the pseudo-beta-l globin gene. The third (300 bp), 3-4 kb 3' to the pseudo-beta-l-globin gene, has the opposite orientation. Their orientations relative to five previously characterized Alu sequences from this cluster have been established. One of these, 2.5 kb 5' to the epsilon-globin gene, was shown by Southern blot hybridization to be similar but not identical to other family members, whereas the region separating it from a neighbouring inverted repeat is not widely distributed in the human genome. PMID- 6271283 TI - Specific membrane receptors for diferric-transferrin in cultured rat skeletal myocytes and chick-embryo cardiac myocytes. AB - A classical approach was used to determine whether myocyte cultures exhibited diferric-transferrin binding phenomena consistent with the presence of specific, high-affinity membrane receptors. Experiments were performed using a continuous cell line, designated L-6, derived from rat skeletal muscle, as well as primary cultures of chick-embryo cardiac myocytes. Rat transferrin isolated from pooled serum was used in experiments involving L-6 cells, and ovotransferrin isolated from hen's egg white was used with the chick-embryo cardiac myocytes. The data from equilibrium binding experiments, corrected for nonspecific binding, and analyzed by Scatchard analysis indicated that there were approximately 2 x 10(5) transferrin receptors per L-6 myocyte and 2 x 10(4) ovotransferrin receptors per cardiac myocyte present, under the conditions used for the equilibrium binding experiments. Whereas the L-6 myocytes grew exponentially under the assay conditions, the cardiac-myocyte cultures were in a non-dividing state. It is thought that the differences in receptor number per cell reflect changes arising form the differing ion demand made by the cells, under these two growth conditions. It is clear that myocytes acquire iron from diferric (ovo)transferrin in a process that involves high-affinity, specific binding to membrane receptors. PMID- 6271284 TI - Investigation of electron-transfer reactions of proteins by electrochemical methods. PMID- 6271285 TI - A transport model for the (Na+/K+)ATPase in liposomes including the (Na+/K+) channel function. AB - (Na+/K+)ATPase liposomes of various degrees of reconstitution are formed by varying the amount of phosphatidylcholine added to the soluble (Na+/K+)ATPase before vesicles are formed by cholate removal. In the presence of ATP, the reconstituted sodium pump effectuates (Na+/K+) antiport. In the absence of ATP, the reconstituted sodium pump forms a (Na+/K+) channel. The stable plateaus formed by (1) the active Na+ transport, (2) the active K+ transport, (3) the 'passive' Na+ flux, and (4) the 'passive' K+ flux are determined in the optimally and the partially reconstituted liposomes. The activities of all four vectorial functions vary in a tightly correlated fashion, suggesting that they are mediated by the same transport-active configuration of (Na+/K+)ATPase. A transport model which includes the active and the passive (Na+/K+) fluxes mediated by the sodium pump in liposomes is outlined. PMID- 6271286 TI - [Spin probe study of the effect of cholera toxin on enterocyte brush border membranes]. AB - The physical state of the enterocyte brush border membranes during stimulation of cholera diarrhea was examined by electron paramagnetic resonance with the use of spin probes localized in different areas of the lipid bilayer and the spin label covalently bound with protein SH-groups. In vivo experiments have disclosed a slight increase in bilayer fluidity during cholera intoxication. In experiments in vitro the binding of cholera toxin with the enterocyte brush border membranes on addition of 3',5'-AMP (up to 5 X 10(-5) M) did not affect the physical state of the lipid of protein areas of the membrane. PMID- 6271287 TI - [Effect of cAMP on hexokinase and glucose-6-phosphate dehydrogenase isoenzymes in different white rat tissues]. AB - Preincubation of liver and skeletal muscle cytosol obtained from Wistar rats in the presence of cAMP, ATP and MgCl2 inhibited hexokinase. The decrease in the total enzymatic activity was a consequence of inhibition of types 2 and 3 hexokinase. The activity of type 1 hexokinase was unchanged. Under the similar conditions there was a lowering of the total activity of glucose-6-phosphate dehydrogenase which arose from inhibition of types 1 and 2 of the enzyme. The activity of other glucose-6-phosphate dehydrogenase isozymes remained unchanged. Preincubation of adrenal supernatant with cAMP, ATP and MgCl2 did not alter the activity of both enzymes. This is likely to be due to the absence of the respective cAMP-dependent proteinkinases and to the features of the isozyme spectrum. The results obtained indicate that hexokinase and glucose-6-phosphate dehydrogenase can be attributed to the group of enzymes that are regulated by the "phosphorylation-dephosphorylation" mechanism. PMID- 6271288 TI - [Synthesis of a high-capacity immunosorbent based on a cellulose suspension]. AB - A method is suggested for preparation of an immunosorbent on the basis of cellulose suspension. Antigen was coupled to periodate-oxidized cellulose via aldehyde groups. Optimal conditions (time of oxidation, amount of an oxidant, quantity of protein antigen added) for immunosorbent synthesis were determined. The amount of antibodies bound to the immunosorbent was approximately equal to the weight of the immunosorbent (up to 950 mg of antibodies per 1 g sorbent). Thus each molecule of antigen coupled to cellulose bound 5-8 molecules of antibodies. PMID- 6271290 TI - [Characteristics of cyclic nucleotide phosphodiesterase and calmodulin in the small intestinal mucosa of rabbits]. AB - Some aspects of the functioning of phosphodiesterase of cyclic nucleotides and calmoduline of the small intestinal mucosa were examined. It was shown that Ca2+ ions virtually have no effect on phosphodiesterase activity of the mucosal homogenate. Cytosol chromatography on a DEAE-cellulose column makes it possible to find an activating effect of calmoduline on phosphodiesterase. Minor quantities of this form of the enzyme might activate the total rate of cAMP hydrolysis with Ca2+ ions for not more than 10%. The author describes calmoduline from the intestinal mucosa as a low-molecular, thermostable protein of acid nature, whose effect can be detected only in the presence of Ca2+ ions. PMID- 6271289 TI - [Effect of benz(a)pyrene and its non-carcinogenic analog 1,2-benzopyrene on the Na, K-ATPase activity of homogenates of different mouse organs and rat heart membrane preparations]. AB - The carcinogenic agent benz(a)pyrene inhibits Na, K-ATPase activity of organ homogenates where it is capable of inducing tumors. It causes no significant changes in target organ homogenates. PMID- 6271291 TI - [Effect of ACTH4-7 and lysyl-vasopressin on the activity of a generator of pathologically enhanced excitation]. AB - It was shown in experiments on random-bred rats that ACTH4-7 and lysine vasopressin promoted the maintenance of the activity in the generator of pathologically enhanced excitation (PEE) created in the anterior horns of the lumbosacral segments of the spinal cord. The effect described manifested clinically in the prolongation of the hypertonus maintenance, and increased electrical activity in the muscles of the appropriate hind limb as compared with the same parameters in the control animals. The effect seen is discussed from the standpoint of the influence of the peptides under consideration on the generator of PEE as on a peculiar form of the pathological memory. PMID- 6271292 TI - [Effect cold adaptation to the ouabain-sensitive component of kidney respiration in rats]. AB - In the course of intermittent cold adaptation (0 degrees C, for 16 weeks) the rats demonstrated an increase in the rate of oxygen consumption by a suspension of kidney pieces. The increase of oxygen consumption by 60% is ouabain-sensitive. Together with the data on Na, K-ATPase activation this indicates the main role of the Na-pump in the increased respiration of the kidneys on cold adaptation. PMID- 6271293 TI - [Cytoplasmic membrane damage and thromboplastinemia in hypercholesterolemia]. AB - Experiments on 67 rabbits were performed to examine the injured external cell membranes of the vascular wall, appearing in the blood flow in the course of the development of alimentary hypercholesterolemia. The time of the appearance in the blood plasma of the external cell membranes was judged from the activity of their specific marker, 5'-nucleotidase. An abrupt increase in 5'-nucleotidase activity was disclosed in the blood plasma at the height of hypercholesterolemia. The latter circumstance served as an objective criterion for injury to ther external cell membranes and might be regarded as a risk factor in thrombus formation. PMID- 6271294 TI - [Activation succinate dehydrogenation in rat liver by noradrenaline, cAMP and acute cooling]. AB - Administration of noradrenaline in a dose of 11 mumol/kg activates succinate dehydrogenase (SDH) in the rat liver. On incubation of liver homogenate with noradrenaline (10(-6) M), cAMP (10(-6) M) and NaF (10(-2) M), a nonhormonal activator of adenylate cyclase, SDH was found to be activated by 20--30% in all the cases. The effects of noradrenaline and cAMP did not sum up. It seems likely that SDH activation by catecholamines is mediated by cAMP. Acute cooling (--17 degrees C) stimulates SDH via endogenous catecholamines, since this effect is completely reversed by the beta-adrenolytic inderal. PMID- 6271295 TI - [Effect of cAMP of glycolysis and glycogenolysis in the liver and adrenals of white rats]. AB - The rates of glycolysis and glycogenolysis an the rate of lactate formation from glucoso-6-phosphate (G-6-Ph) in the liver were reduced during stress (starvation). On the contrary, these activities in the adrenals were increased. The rates of lactate formation from fructose diphosphate remained unchanged in both organs. The results obtained attest to the inhibition in the liver and activation in the adrenals of phosphorylase, hexokinase and phosphofructokinase. The degree of hexokinase inhibition in the liver depended on the presence of cAMP, ATP and MgCl2 in the incubation medium and was a consequence of enzymatic phosphorylation. Unlike 2', 3'-AMP, the inhibitory effect of CAMP was highly specific. The protein inhibitor of protein kinase completely reversed the inhibitory effect of cAMP on hexokinase. In the adrenals, cAMP slightly increased the rates of glycolysis and lactate formation from G-6-Ph because of allosteric effects of cAMP. The activation rather than inhibition of glycolysis in the adrenals during stress is probably caused by the absence in this tissue of cAMP dependent protein kinase which phosphorylates hexokinase. PMID- 6271296 TI - [Effect of peritol on the activity of the hypothalamo-hypophyseo-adrenal system]. AB - A study was made of the activity of the hypothalamo-hypophyseo-adrenal system under the effect of peritol (ciprohepatadin), an antagonist of serotonin receptors. Experiments were carried ot on male rats in which the blood content of corticosterone, aldosterone and ACTH was determined 3, 6 hours and 14, 20 and 30 days after peritol administration (twice a day). Pronounced inhibitory effect on the hypothalamo-hypophyseo-adrenal system was found to be attainable on a more frequent exposure (every 3 hours) to peritol with a purpose of maintaining the optimal drug concentration in various body tissues. The latter circumstance gives rise to peritol effect which is realized through both the central mechanisms (ACTH content) and as a result of direct action on steroid output. PMID- 6271297 TI - [Effect of manganese and verapamil on electrical and contractile responses of pulmonary artery smooth muscle induced by noradrenaline and histamine]. AB - Action of noradrenaline and histamine on the resting potential, membrane resistance and contractility of rabbit pulmonary artery muscle cells was investigated in normal and Ca-blockers (manganese and verapamil)-containing Ringer-Lock solutions. It was shown that catecholamine and histamine induced depolarization by different mechanisms. Thus, noradrenaline action is accounted for by the decreased membrane permeability to potassium ions, while the histamine induced depolarization is a consequence of sodium and, probably, chlorine permeability. The contraction induced by the transmitters is activated primarily by the extracellular calcium ions entering the cells by two ways: via chemosensitive Ca-channels activated by adrenergic and histaminergic receptors or via potential-dependent slow Ca-channels activated by the transmitter-induced membrane depolarization. It is not excluded that during activation of muscle cells by the transmitters part of calcium is release from both intramembrane and intracellular stores. PMID- 6271299 TI - [Age and ultrastructural features of adrenal capillaries in rats following ACTH administration and in insulin shock]. AB - Electron microscopy was used to study age-associated features of blood capillaries of adrenal cortical substances and adrenal medulla in adult and old rats during ACTH administration and insulin shock. It has been revealed that as a result of an increase in adrenal function of the adult rats, endotheliocytes of cortical substance and medulla developed ultrastructural changes promoting the intensification of transcapillary metabolism. The old rats showed pathological changes in the endotheliocyte ultrastructure, responsible for the development of capillary insufficiency of the adrenals during stimulation of their secretory activity. PMID- 6271298 TI - [Mechanism of action of collagen on processes of regeneration]. AB - The effect of collagen, its hydrolysate and glycin on metabolism of cyclic nucleotides in wound tissue was studied in experiments on animals. Collagen was found to reduce the cAMP level in muscles of the wound fundus, while the concentration of cGMP remained unchanged. Collagen hydrolysate induced unidirectional changes in cyclic nucleotides, whereas glycin opposite ones. The basal activity of cAMP phosphodiesterase was not changed. The mechanism of the stimulatory effect of collagen on wound healing is discussed. PMID- 6271300 TI - [Response of identified mollusk neurons to treatment with certain metabolic regulators during different oxygen supply conditions]. AB - The time course of redox systems of some identified Helix pomatia neurons and their bioelectrical activity during exposure to cAMP and prostaglandin E2 under normoxia and anoxia was studied by means of a complex technique on the basis of contact vital microscopy. It was discovered that neuronal response to the substances was uncertain under varying conditions of neuronal oxygen supply. In the presence of preliminary exposure to cAMP and prostaglandin E2 the character of shifts in redox systems and bioelectrical activity occurring in the neurons during oxygen deprivation was to be changed. PMID- 6271301 TI - [Changes in the levels of 3',5'-cAMP and 3',5'-cGMP in brain and heart tissues after exposure to toxins of Buthus eupeus scorpion venom]. AB - Toxins isolated from Buthus eupeus scorpion stimulated the generation of cAMP and cGMP in mouse cerebellar and forebrain slices, as well as in isolated guinea-pig heart. Pharmacological analysis of neurotropic and cardiotropic effects of toxins has ascertained the relationship between the increased level of cyclic nucleotides and catecholaminergic and cholinergic effects of toxins. PMID- 6271302 TI - [Early changes in retinal 5'-nucleotidase activity in hereditary retinal degeneration]. AB - Activity of soluble cGMP phosphodiesterase (PDE) and of two membrane enzymes, 5' nucleotidase and Na,K-ATPase, was studied in the developing retina of rats with inherited retinal degeneration. It was found that by day 10 of life, the content of 5'-nucleotidase in the afflicted rats was significantly reduced as compared with controls. This difference was unchanged throughout the subsequent animals' life. Na,K-ATPase activity in the afflicted and normal animals was the same. Within the first 45 days of life, PDE calculated with respect to the rhodopsin content was not different as regards both the afflicted and normal rats. When calculated with respect to protein, the changes in PDE corresponded with the reported data. The data obtained allowed a suggestion to be made that changes in 5'-nucleotidase in inherited retinal degeneration are disease-specific. They are accounted for by changes in the enzymes of nonphotoreceptor retinal membranes. The changes in PDE may be regarded as secondary, correlating with variation in the number of the photoreceptor membranes. PMID- 6271303 TI - [Factors modulating production of antibodies against Clostridium perfringens in mammals]. AB - Immunogenic properties of Clostridium perfringens type A toxoid preparations obtained by different methods are described. As regards immunogenicity for guinea pigs and man, toxoid obtained by the detoxification of preliminarily purified alpha-toxin (experimental toxoid) compares favourably with preparations obtained by the detoxification of alpha-toxin in a culture fluid. It was shown in experiments on guinea pigs that immunogenicity of experimental toxoid rises with the increase in the degree of purification of alpha-toxin used for detoxification. The least purified preparations turned out to be the most immunogenic for mice. In Cl. perfringens culture fluid, factor (factors) was found and separated, which reduced the immunogenicity of the respective toxoid in experiments with immunization of guinea pigs and increased it in mice. Activation of antibody-formation was disclosed to be nonspecific. The existence of this factor(s) should be taken into account in the development of vaccines and methods for evaluating their immunogenic properties. PMID- 6271304 TI - [Ability of components of pertussis bacteria to induce delayed-type hypersensitivity to DNA]. AB - Bordetella pertussis vaccine contains antigens which are capable to evoke delayed type hypersensitivity (DTH) reactions to deoxyribonucleic acid, except specific antipertussis response in mice. This phenomenon signifies the possibility of the autoimmune process. The purified derivatives of B. pertussis maintain specific activity but do not stimulate the expression of DTH to DNA. PMID- 6271305 TI - [Gene mutation induction by benz(a)pyrene following metabolic activation]. AB - Chinese hamster cells were made use of to study the effect of low doses of benz(a)pyrene on gene mutations during metabolic activation. The HGRPT mutant clones were selected on the basis of their resistance to 8-azaguanine. The raising of benz(a)pyrene concentration (from 5 g/ml) was followed by the enhanced mutagenic effect which was approximately 7-10 times greater than the spontaneous level at a concentration of 20-25 micrograms/kg. PMID- 6271306 TI - [Cell cycle and proliferative pool of human tumor strains transplanted into athymic mice]. AB - Autoradiography was used to study the cell cycle periods and proliferative pools of human tumor strains (Ewing's sarcoma, melanoma, Wilms' tumor, renal carcinoma) transplanted in nude mice. The duration of cell cycles was found to be as follows: 22.2 h for Ewing's sarcoma; 16.3 h for melanoma; 22.8 h for Wilms' tumour; 28.2 h for renal carcinoma, with the proliferative pools being equal to 90.1; 89.9; 71.7 and 21%, respectively. The shorter duration of the melanoma strain cycle is primarily accounted for by the reduced S phase. PMID- 6271308 TI - [Mechanisms of the toxic effect of benz(a)pyrene and its derivatives in tissue culture]. AB - Study in cell cultures of toxic action and metabolism of benz(a)pyrene and its derivatives substituted in the 6th position has shown that the toxic effect of the compounds depends on the substituent moiety. No correlation has been disclosed between toxicity and metabolism intensity. It is suggested that toxic action of benz(a)pyrene and its 6-substituted is determined by epoxide formation in the 4th-5th position. PMID- 6271307 TI - [Possible participation of endogenous MMTV virus in chemical carcinogenesis of mouse mammary glands]. AB - A study was made of the effect of immunization with formalinized MMTV on mammary tumor incidence in BALB/c mice exposed to dimethylbenzanthracene. In mice immunized with MMTV, the incidence of mammary tumours was significantly lower and the latent period longer as compared with the control group immunized with MULV. The indirect fixed immunofluorescence technique has shown that tumour cells in both groups express gp52 and p27 antigens. The possible involvement of endogenous MMTV into chemical carcinogenesis in murine mammary glands is discussed. PMID- 6271309 TI - [Autoradiographic study of hormonal regulation of proliferating golden hamster uterus cells during postnatal ontogeny]. AB - Study of the effects of octestrol, 17-hydroxyprogesterone caproate and tamoxifen on uterine cell proliferation in golden hamsters aged one to twenty days has shown the existence in the uterine epithelium of two cell populations with varying sensitivity to sex hormones. The cells of uterine glands being formed by day 12 of postnatal ontogenesis were found to be insensitive to the proliferation inhibiting effect of progesterone and little sensitive to the proliferation stimulating action of the estrogen. The epithelial cells lining the uterine cavity show a high sensitivity to sex hormones. The absence of changes in proliferative activity of the epithelial cells exposed to tamoxifen allows a suggestion that ontogenetic changes in the proliferative activity of the uterine cells do not depend on the presence of endogenous hormones. PMID- 6271310 TI - Long-term suspension cultures of human cord blood myeloid cells. AB - A method is described that allows for the routine long-term (greater than 3 mo) growth of normal, immature human myeloid cells in liquid suspension culture. The techniques employ cell separation, utilization of special growth conditions (RPMI with hydrocortisone and vitamin D), and cord blood as the source of leukocytes. These cultures are composed predominantly of immature myeloid cells that have grown for over 3 mo in culture but eventually terminate as differentiated, mature granulocytes and monocyte-macrophages. Application of these techniques to other sources of fresh human leukocytes (bone marrow and adult peripheral blood) resulted in only short-term proliferation of myeloid cells. The techniques described can be used for the routine expansion of immature myeloid and monocytoid cell populations, particularly from newborns, and should be useful for systematic studies of normal myeloid-monocytoid cell growth and differentiation and providing normal control cells in studies employing human leukemic myeloid cells. PMID- 6271311 TI - Opsonized zymosan-stimulated granulocytes-activation and activity of the superoxide-generating system and membrane potential changes. AB - Phagocytic cells generate superoxide in response to stimulation by opsonized particles. A continuous assay for opsonized zymosan-stimulated granulocyte superoxide production shows that there is a lag time between the addition of particles and the onset of detectable superoxide production. Superoxide production is preceded by membrane potential depolarization. Neither superoxide production nor membrane depolarization occurs in granulocytes from patients with chronic granulomatous disease. The extent of activation by opsonized zymosan is affected by the dose of zymosan from 0.5 to 4.5 mg/ml, but the time necessary for activation (lag time) is not. Similarly, the extent of depolarization but not the time necessary for attaining maximum depolarization is concentration-dependent. Effects of temperature, divalent cations, 2-deoxyglucose, cyanide, and N-ethyl maleimide on superoxide production are similar for granulocytes treated with soluble stimuli and with opsonized zymosan. Thus, zymosan stimulates granulocytes to generate superoxide and undergo membrane depolarization in a manner similar to that elected by soluble stimuli. PMID- 6271312 TI - Spin label study of erythrocyte deformability I. Electron spin resonance spectral change under shear flow. AB - A spin labeling method in electron spin resonance spectroscopy (ESR) is applied for the first time to study the deformability of human red blood cells (RBC). ESR measurements of a RBC suspension incubated with a fatty acid spin label were performed, using a narrow-gap flat ESR sample cell under various flow shear stresses (tau). Remarkable changes were observed in ESR spectra with tau, indicating that RBC are oriented in such a way that the greater part of the membrane surface is aligned parallel to the ESR cell walls. The diamide-treated, hardened RBC, in which the biconcave discoid shape remains intact under no shear stress, exhibit a smaller ESR spectral change with tau than the intact, demonstrating that the present method can be used to assess the deformation of RBC occurring with flow orientation. In particular, the relative amplitude of an ESR difference spectrum may be used as a measure of the elongation of RBC. The conclusion is further supported by experiments using glutaraldehyde-treated or heat-denatured RBC. All these ESR results are in good agreement with the corresponding results obtained by several different methods. The present spin labeling technique is thus proven to be applicable for evaluating RBC deformability. PMID- 6271313 TI - Alpha-adrenoreceptor-related dissociation constants and intrinsic efficacies of stereoisomers of epinephrine. AB - THe alpha-adrenoreceptor-related dissociation constant and intrinsic efficacy of the stereoisomers of epinephrine were determined on the rabbit aortic strip. (-) Epinephrine showed higher affinity than (+)-epinephrine or the desoxy analogue epinine. Efficacy of (+)-epinephrine relative to (-)-epinephrine was low (relative efficacy, er = 0.44). Epinephrine had about the same affinity and efficacy as (+)-epinephrine. When the alpha-adrenoreceptors were protected against dibenamine by an equimolar concentration of the isomers, (-)-epinephrine was found to be more effective than (+)-epinephrine or epinine. These results suggest that the stereoisomers of catecholamines differ not only in affinity but also in intrinsic efficacy. PMID- 6271314 TI - Detection of a togavirus-like agent in bovine leukocytes. PMID- 6271315 TI - Experiments with a combined vaccine for poultry. PMID- 6271316 TI - Effects of some pesticides on enzyme activities in an organic soil. PMID- 6271317 TI - A comparison of cardiac reactivity and beta-adrenoceptor number and affinity between aorta-coarcted hypertensive and normotensive rats. AB - 1 The effects of noradrenaline (NA) and isoprenaline on isolated atria from aorta coarcted hypertensive rats (AHR) at early (6 day) and chronic (28 day) stages of hypertension were studied and compared with time-matched, sham-operated, normotensive rats (SNR). The number and affinity of beta-adrenoceptor ((-)-[3H] dihydroalprenolol binding sites) were also studied in cardiac membranes prepared from these animals. 2 Six and 28 days after complete ligation of the abdominal aorta between the two renal arteries, rats became hypertensive with significantly greater arterial blood pressures than time-matched SNR. 3 At both stages of hypertension, the atrial inotropic or chronotropic effects of NA and isoprenaline from hypertensive rats were similar to time-matched SNR. Moreover, no differences in atrial reactivity were observed between the early and chronic stages of hypertension. 4 Irrespective of the stage of hypertension, cardiac membranes from the AHR contained the same number of beta-adrenoceptors as time-matched SNR. In addition, the receptor affinity for the radioligand within each group was equivalent. However, the chronic stage hypertensive rats and their time-matched controls contained fewer beta-adrenoceptors and these receptors had greater affinity for the radioligand when compared with cardiac membranes from rats at the early stage of hypertension and their controls. 5 The observed equivalent chronotropic and inotropic responses to NA and isoprenaline between the hypertensive and normotensive rats in both stages of hypertension may be explained in terms of similar receptor number and receptor binding affinity. 6 The reduced number of beta-adrenoceptors with greater binding affinity in day 28 normotensive or hypertensive rats may be a compensatory mechanism for these animals to maintain normal cardiac function with increasing age. PMID- 6271318 TI - Sympathetic mechanisms in diet-induced thermogenesis: modification by ciclazindol and anorectic drugs. AB - 1 The sympathetic noradrenergic activation of brown adipose tissue and the biochemical mechanisms involved in diet-induced thermogenesis were studied in rats. 2 A close correlation was found between brown adipose tissue Na+, K+ adenosinetriphosphatase (Na+, K+-ATPase) activity in vitro and in vivo measurements of resting oxygen consumption (VO2). The effects of noradrenaline on in vitro NA+, K+-ATPase activity in brown adipose tissue and in vivo VO2 could be mimicked by a variety of agents. These included beta-adrenoceptor agonists and agents known to induce the release of noradrenaline or inhibit the noradrenaline uptake process. The pharmacological evidence suggests that dopaminergic mechanisms may also be involved in the control of thermogenesis. 3 Amphetamine did not increase VO2 in rats without causing associated increases in locomotor activity. Ciclazindol at doses of 3-30 mg/kg intraperitoneally increased VO2 but did not appear to increase locomotor activity or evoke any other signs of CNS stimulation including lengthening of time to sleep onset or stereotypy. Separation of metabolic and CNS effects occurred only at the lowest dose of mazindol used (0.3 mg/kg i.p.). These results are probably a reflection of (a) the relative abilities of these drugs to inhibit brain and brown adipose noradrenaline uptake processes and (b) the relatively high accumulation of ciclazindol in brown adipose. 4 Of the drugs tested, only ciclazindol was a more potent inhibitor of the noradrenaline uptake system in brown adipose tissue (BAT) than in brain. Kinetic analysis also revealed that the actions of ciclazindol on the NA uptake system and Na+, K+-ATPase in BAT differed from those of mazindol. 5 These findings suggest that ciclazindol may produce an energy wasting effect in rodents without causing overt CNS stimulation; the implications of these findings in terms of human obesity are discussed. PMID- 6271319 TI - The action of loperamide in inhibiting prostaglandin-induced intestinal secretion in the rat. AB - 1 The mechanisms by which loperamide inhibits the intestinal secretion induced by prostaglandin E2 were investigated in rat jejunum. 2 In vivo loperamide prevented prostaglandin-induced fluid secretion but did not reduce the associated rise in the transintestinal potential difference. 3 In intestinal sheets the electrical response to prostaglandin E2 was enhanced in the presence of loperamide. 4 The ionic basis of these changes was determined by measuring Na+ and Cl- fluxes across intestinal sheets. Loperamide did not reduce the prostaglandin-induced increase in net Cl- secretion, although it prevented the inhibition of mucosal-to serosal Na+ movement. 5 Loperamide does not alter cyclic adenosine 3',5' monophosphate (cyclic AMP) levels by a direct action at the enterocyte, since in isolated enterocytes neither basal nor prostaglandin-stimulated cyclic AMP levels were affected by the drug. PMID- 6271320 TI - Two types of gamma-aminobutyric acid receptor on embryonic sensory neurones. AB - 1 Embryonic sensory neurones of the chick grown in dissociated cell culture respond to application of low concentrations of gamma-aminobutyric acid (GABA) with a change in resting membrane resistance (R(in)) and/or a change in action potential duration (APD) (Dunlap & Fischbach, 1978; Choi & Fischbach, 1981). Intracellular microelectrode recording techniques were employed to determine if these two effects are mediated by the same, or different, GABA receptors.2 Cells responded, for the most part, with a change in either R(in) or APD, but 10% of the cells exhibited both effects. In the latter cells the two responses were clearly distinguishable as discussed below.3 The proportion of neurones exhibiting a GABA-induced decrease in R(in) declined during the first week in vitro while the proportion exhibiting a decrease in APD increased during that time.4 The two effects were pharmacologically distinct. Muscimol, a GABA analogue, produced only the change in R(in) (ED(50) = 5.5 muM) while baclofen, another analogue of GABA, produced only the change in APD (ED(50) = 1 muM). The analogues were approximately equipotent with GABA. Bicuculline, a GABA antagonist, blocked the muscimol-induced change in R(in) (but not the baclofen induced change in APD) in a dose-dependent fashion with an ID(50) = 0.7 muM.5 The time courses of the two effects were different. The change in APD resulting from a brief application of GABA (or baclofen) was prolonged relative to the rapid return to control associated with the GABA- (or muscimol-) induced change in R(in).6 Desensitization of the two responses exhibited separate time courses. In the continual presence of the agonists, GABA- and muscimol-induced decreases in R(in) completely desensitized in ca. 10 s while GABA- and baclofen-induced decreases in APD persisted undiminished throughout a prolonged (1 min) application of the drugs and returned to control only after cessation of application.7 It is concluded that embryonic chick sensory neurones in culture exhibit two types of GABA receptor that differ in their functional and pharmacological properties. Implications of these results are discussed. PMID- 6271322 TI - In vivo receptor binding of the opiate partial agonist, buprenorphine, correlated with its agonistic and antagonistic actions. AB - 1 In order to gain more insight into the mechanisms behind the actions of opiate partial agonists, an analysis of the dual agonist/antagonist properties of the partial agonist, buprenorphine, was made in conjunction with in vivo binding studies on the drug in the rat. 2 Buprenorphine revealed a bell-shaped dose response curve for antinociception peaking at approx. 0.5 mg/kg subcutaneously. It antagonized morphine antinociception at doses which normally have agonistic effects and produced maximum antagonistic effects at doses above those having prominent agonistic activity. The withdrawal precipitating potency of buprenorphine as measured in highly morphine-dependent rats was present at doses above those having agonistic activity. The entire dose-response curve for buprenorphine was shifted symmetrically to the right by the opiate antagonist, naltrexone. 3 The dose-dependent occupation of receptors in vivo by buprenorphine seemed to be almost complete over the agonist dosage range; almost no further receptor occupation over the antagonist range was seen. 4 The possibility is discussed that site-to-site receptor interactions leading to cooperativity of effect may be the best explanation of these results. PMID- 6271321 TI - Receptor mechanisms for 5-hydroxytryptamine in rabbit arteries. AB - 1 Previous investigations into the vascular actions of biogenic amines implicated in migraine have shown that the contractile effects of both 5-hydroxytryptamine (5-HT) and noradrenaline (NA) in the rabbit ear artery are mediated by a direct sympathomimetic action at alpha-adrenoceptors, while in the rabbit aorta, 5-HT and NA act on pharmacologically distinct receptors. The purpose of the present investigation was to determine whether the absence of 5-HT receptors in rabbit ear arteries is characteristic of distributing arteries in general, or is confined to particular regional circulations.2 Agonist-antagonist interactions were studied in various rabbit vascular preparations (common carotid, external carotid and femoral arterial strips, and perfused ear arteries) by determining pA(2) values for pizotifen and phentolamine against 5-HT- and NA-induced contractile responses.3 In common carotid and femoral arteries, pizotifen was a potent competitive antagonist of 5-HT, but weak against NA. The converse applied to phentolamine. In external carotid and ear arteries, pizotifen was a weak competitive antagonist of both 5-HT and NA, whereas phentolamine was a potent competitive antagonist of both. Cocaine did not influence pA(2) values against NA.4 5-HT and NA were of similar potency in common carotid and femoral arteries, but 5-HT was much less potent than NA in external carotid and ear arteries.5 The results indicate that rabbit common carotid and femoral arteries contain both D type 5-HT receptors and alpha-adrenoceptors, as does the aorta. However, external carotid arteries, like ear arteries, do not contain specific 5-HT receptors. The action of 5-HT in the external carotid artery is mediated by alpha-adrenoceptors; this is a direct sympathomimetic action since it was not inhibited by cocaine or reserpine-pretreatment.6 The absence of 5-HT receptors in the rabbit extracranial circulation may limit the usefulness of this species as a model for research relating to migraine. PMID- 6271324 TI - New modalities in cancer treatment: heavy charged particles. AB - Heavy charged particles represent the ultimate that the physicist can contribute to the development of radiation sources for therapy. Of the heavy charged particles, protons are the least expensive to accelerate and can be manipulated to give a sharply defined high-dose volume with a rapid fall-off of dose outside the target area. The biological properties of protons do not differ significantly from X or gamma rays. Negative pi mesons require an elaborate accelerator for their production. Pions offer the possibility of concentrating energy, some of it densely ionizing with a reduced oxygen enhancement ratio and elevated biological effectiveness within the designated tumour volume, while minimizing the dose of sparsely ionizing radiation to the normal tissues traversed. High-energy heavy ions offer the greatest flexibility and allow localized dose distributions and also, with the higher Z particles, a substantial reduction of the oxygen enhancement ratio can be achieved. The ultimate choice of particle depends upon what turns out to be the most important factor in radiotherapy--an improved localization of dose or a reduction in the dependence of cell killing on the presence of molecular oxygen. PMID- 6271323 TI - The calcium antagonistic effects of cyproheptadine on contraction, membrane electrical events and calcium influx in the guinea-pig taenia coli. AB - 1 The ability of cyproheptadine (Cph) to inhibit membrane translocation of calcium in smooth muscle was investigated by studying the drug's action on contraction, electrical activity and calcium influx in the guinea-pig taenia coli.2 Cph >/= 10(-6)M reduced the amplitude of normal spontaneous contractions and concurrently decreased the number of action potentials occurring with each slow-wave of depolarization (sucrose-gap recordings). These inhibitory effects of Cph were antagonized by increasing the medium [Ca] three fold to 7.68 mM.3 Intracellular recordings showed that Cph >/= 2 x 10(-6)M decreased the amplitude and extended the duration of the action potential. These effects were only partially reversible in normal medium whereas large overshooting action potentials were again seen in 7.68 mM Ca medium.4 High frequency mechanical activity was produced by inclusion of veratridine 5 x 10(-6)M in the perfusate. Low concentrations of Cph (>/= 10(-7)M) reduced the amplitude of such contractions at a faster rate than they did normal spontaneous contractions.5 At concentrations between 10(-7) and 10(-6)M, Cph fully reduced the tonic component of contractions elicited in 112 mM isotonic KCl whilst having little or no effect on either (i) the initial phasic KCl contraction or (ii) the ;repolarization contracture' normally produced on wash-out of the KCl or (iii) the spontaneous contractions before and after KCl treatment. In contrast, at Cph 2 x 10(-6)M, the repolarization contracture, as well as the isotonic KCl contraction, was totally blocked whereas spontaneous contractions were still unaffected. Progressively higher Cph concentrations inhibited all components of this contractile cycle.6 Dose-response curves for the rate of drug-induced relaxation of tonic contractures produced in hypertonic 42.7 mM high-potassium medium, showed the calcium antagonistic potency of Cph to be intermediate between that of chlorpromazine and D600. The minimum Cph concentration for effect lay between 1 and 5 x 10(-7)M, and the effects of Cph 2 x 10(-6)M (approximately the ID(50)) were totally antagonized by 12.8 mM Ca.7 By means of a lanthanum wash procedure, Cph >/= 2 x 10(-6)M was found to decrease the (45)Ca uptake occurring into strips of taenia coli in normal medium, although the maximum effect (at Cph 10(-5)M) amounted to only 25% inhibition of the uptake occurring into control strips (also found with D600). The increased uptake occurring in hypertonic 44.7 mM high potassium medium was inhibited in a dose-dependent manner by Cph 1 x 10(-7)M.8 The results are consistent with an action of Cph in reducing the flow of Ca(2+) through voltage-dependent Ca channels in the smooth muscle cell membrane. It is suggested that the interaction of Cph molecules with such sites is dependent upon membrane potential as well as drug concentration. PMID- 6271325 TI - Ultrasound in hepatoma. PMID- 6271326 TI - Therapeutic adrenal venous infarction in ACTH-dependent Cushing's syndrome. PMID- 6271327 TI - Thiol radioprotection in vivo: the critical role of tissue oxygen concentration. PMID- 6271328 TI - Toxic hepatis during ketoconazole treatment. PMID- 6271329 TI - Shipyard eye. PMID- 6271330 TI - Hepatitis and ketoconazole therapy. PMID- 6271331 TI - Time for a change of MIND. PMID- 6271332 TI - The effects of neuroleptics on the GABA receptor of cat primary afferent neurons. AB - Butyrophenones (haloperidol and pimozide) at low concentrations (0.05-1.0 micro M) inhibited the GABA-induced depolarization of cat primary afferent neurons, while at high concentrations (greater than 10 micro M) they enhanced the GABA depolarization. These actions of the butyrophenones were not accompanied by any measurable change in the dissociation constant of the GABA-GABA receptor interaction, whereas their inhibitory and facilitatory influences on the GABA depolarization were associated with reduction and increase in the cell membrane resistance, respectively. Further analysis showed that the reduction of membrane resistance by low concentrations of butyrophenones was brought about by an increased sodium and potassium conductance and that the increase in membrane resistance by a high concentration of these drugs was caused by a reduced sodium and potassium conductance. In contrast to the butyrophenones, a typical phenothiazine derivative (chlorpromazine) at concentrations of 0.1-100 micro M did not affect the GABA-depolarization. The results suggest that butyrophenones do not mimic the action of GABA as originally proposed by Janssen, but alter the GABA-induced depolarization indirectly by modifying the electrically excitable portion of the cell membrane. Chlorpromazine, a phenothiazine, on the other hand, has no recognizable effects on either the GABA-receptor membrane or the electrically excitable membrane. PMID- 6271333 TI - Regulation of GABA receptor binding to synaptic plasma membrane of rat cerebral cortex: the role of endogenous phospholipids. AB - Triton X-100 treatments produced an extensive depletion of proteins and phospholipids and a marked increase of [3H] GABA binding on synaptic plasma membranes (SPM). Maximal [3H]GABA binding was obtained with three Triton X-100 treatments (+ 174% with respect to control). Phospholipase C, which removes only the phospholipid polar head, induces a 40% increase of [3H]GABA binding only after treatments resulting in extensive protein depletion. In reconstitution experiments phosphatidylethanolamine, the largest phospholipid removed, induced a 30-35% inhibition of [3H]GABA binding in Triton X-100 treated membranes; in contrast phosphatidylserine and phosphatidylcholine did not produce significant changes. The reconstitution of phospholipase C-treated SPM preparations with exogenous phosphatidylethanolamine, phosphatidylserine, phosphatidylcholine or phosphoethanolamine and 1,2-dipalmitoylglycerol, products of phospholipase C activity, did not yield significant changes. This evidence, which argues against a direct role of phospholipids on the regulation of GABA binding, should, however, suggest that the GABA binding component of the receptor site is a lipoprotein or a lipid-depending protein. PMID- 6271334 TI - High affinity binding of an enkephalin analog in the cerebral ganglion of the insect Leucophaea maderae (Blattaria). AB - Specific high affinity binding sites for a synthetic enkephalin analog, DALA (D Ala2-Met-enkephalinamide) were demonstrated in the cerebral ganglia of adults and nymphs of the insect Leucophaea. In both sexes binding was monophasic and saturable with respect to the concentration of the radioligand used. The same amount of brain tissue from adult females had 30% more binding sites for DALA than that from adult males. By contrast, no sex-related difference in binding site density per mg protein was observed in the brains from immature (nymphal) specimens. The results strongly suggest the presence, in this invertebrate, of opiate receptors that appear to be confined to certain areas of the nervous tissue. PMID- 6271335 TI - Cytoplasmic alkalization reduces calcium buffering in molluscan central neurons. AB - The effect of raised cytoplasmic pH (pHi) on intracellular concentration ([Ca2+]i) transients following calcium influx during membrane depolarization was studied in identified neurons in the abdominal ganglion of Aplysia californica. The pHi was monitored with pH-sensitive microelectrodes. Sea water containing 15 mM NH4Cl at pH 7.7 elevated pHi about 0.35 pH units from the normal level of 7.17. These cells have an estimated buffering power of about 60 mM/pH unit. Calcium influx was elicited by depolarizing pulses under voltage clamp and [Ca2+]i transients were monitored with the photoprotein aequorin or the metallochromic dye arsenazo III. Aequorin photo-emissions increased by 21--131% (mean, 48%) and arsenazo III absorbance changes accompanying depolarization increased by 9--33% (mean, 20%) after 30 min in NH4+, corresponding roughly to a 14% increase in [Ca2+]i transients. Calcium-dependent potassium tail currents following a depolarizing pulse were somewhat slower and 4--91% (mean, 38%) large in NH4+. The magnitude and time- and voltage-dependence of the membrane calcium conductance was studied using calcium tail currents following depolarizing pulses. The calcium current was unaffected by NH4+, so the enhanced [Ca2+]i transients must reflect reduced calcium buffering at high pHi. Either reduced cytoplasmic calcium binding or slowed active extrusion of calcium may be responsible for this effect. PMID- 6271336 TI - Baclofen selectively inhibits excitatory synaptic transmission in the hippocampus. AB - The effect of baclofen was investigated on mossy fiber, Schaffer collateral and perforant path synaptic transmission in hippocampal slices. Baclofen completely inhibits mossy fiber and Schaffer collateral synaptic transmission with an IC50 of 3.8 microM. The lateral perforant path is insensitive to baclofen, while the response in the medial zone was partially blocked. Baclofen does not appear to act in a GABA-like manner. PMID- 6271337 TI - Electrophysiologic determination of projections from ventromedial hypothalamus to midbrain central gray: differences between female and male rats. PMID- 6271338 TI - Inhibition from ventral tegmental area of nucleus accumbens neurons in the rat. AB - The role of the ventral tegmental area (VTA), which is rich in dopamine containing cell bodies, on nucleus accumbens (Acc) neurons was examined. In Acc neurons receiving input from parafascicular nucleus (Pf) of thalamus, VTA conditioning stimulation produced an inhibition of spike generation with Pf stimulation. In contrast, VTA conditioning stimulation did not affect Acc neurons receiving input from limbic structures such as the amygdala nucleus and hippocampus. PMID- 6271339 TI - Melatonin increases cGMP and decreases cAMP levels in rat medial basal hypothalamus in vitro. AB - Measurement of cyclic nucleotide accumulation in rat medial basal hypothalamus (MBH) incubated in vitro with methoxyindoles indicated that melatonin (10(-8) M or greater) increased significantly cGMP and depressed cAMP levels. Only the effect on MBH cAMP was shared by 5-methoxytryptophol which exhibited a greater activity than melatonin. 6-Fluoromelatonin (10(-7) M) increased cGMP, whereas the effect of 6-hydroxymelatonin was not significant. Both 6-fluoro- and 6 hydroxymelatonin (10(-5) M) depressed MBH cAMP accumulation while only 6 fluoromelatonin affected it at 10(-7) M concentrations. These data suggest that melatonin and 5-methoxytryptophol affect differently cyclic nucleotide content of MBH at physiological concentrations. PMID- 6271340 TI - Chloride-dependent stimulation of GABA and benzodiazepine receptor binding by pentobarbital. AB - Pentobarbital reversibly increased Na+-dependent GABA binding to bovine cerebral cortex membranes. This effect was chloride-dependent. Kinetic analysis of [3H]GABA binding revealed an increase of the apparent number of binding sites. Pretreatment of the membranes with Triton X-100 extremely diminished the effect of pentobarbital on both [3H]GABA and [3H]flunitrazepam binding. PMID- 6271341 TI - Reticular formation influence on neuronal transmission from perforant pathway through dentate gyrus. AB - Electrical stimulation of the perforant pathway discharges granule cell synchronously, giving rise to a characteristic evoked potential in the granule cell layer termed here the evoked action potential or EAP. In freely moving rats, we applied 3 pulses of low intensity electrical stimulation to the medullary reticular formation prior to the application of the perforant path pulse. The effect of prior reticular formation stimulation was a marked augmentation of the normal EAP response to the perforant path stimulus. The augmentation was dependent on the behavioral state of the experimental animal (it occurred during slow-wave sleep but not during still, alert behavior) and was eliminated by anesthetic agents. The latency of EAP augmentation effect (minimum effective time interval between application of the reticular formation stimulus and the perforant path pulse) was 13--18 msec. In order to localize the sites in the medullary reticular formation from which EAP augmentation could be elicited, threshold currents for producing the effect were determined during dorso-ventral penetrations of a reticular formation stimulating electrode. EAP augmentation was elicited at low stimulus currents from a relatively broad region of the reticular formation. It was also noted that reticular formation stimulation which produced EAP augmentation always elicited one or more motor responses of the neck, back, face or vibrissae. Subsequent investigation of the pathways underlying these motor responses suggested that the effect of reticular formation stimulation on granule cell excitability was mediated by a polysynaptic pathway, the first segment of which was a projection to cells of nucleus gigantocellularis of the caudal medulla. PMID- 6271342 TI - Sensitization of group IV muscle receptors to bradykinin by 5-hydroxytryptamine and prostaglandin E2. AB - The aim of the study was to find out whether endogenous substances with a presumed sensitizing action on nociceptors alter the chemical excitability of muscle receptors with unmyelinated afferent fibres. In anaesthetized cats, the discharges of single group IV units in response to intra-arterial injections of bradykinin were evaluated quantitatively and the influence of 5-hydroxytryptamine (5-HT) and prostaglandin E2 (PGE2) on the response magnitude determined. Both 5 HT and PGE2 enhanced the bradykinin sensitivity of most of the muscle receptors, i.e. the receptors were sensitized to bradykinin by 5-HT and PGE2. Units that were activated by bradykinin before administration of the sensitizing chemicals showed an increase in response magnitude; in receptors not responding to the standard dose of bradykinin an activation often occurred after chemical sensitization. It is occurred that under the influence of elevated tissue levels of 5-HT and PGE2 the afferent impulse activity induced in group IV muscle receptors by bradykinin will be higher. As the substances used are released together from pathologically altered organs and since many of the group IV muscle receptors are considered to be nociceptive, the chemical sensitization of these receptors to the algesic agent bradykinin might play a role in the production of pain in an inflamed or injured muscle. PMID- 6271343 TI - Neurophysiological consequences of presynaptic receptor activation: changes in noradrenergic terminal excitability. AB - Experiments were carried out to explore the view that activation of presynaptic receptors on the terminals of noradrenergic neurons is accompanied by alterations in their excitability to direct electrical stimulation. Antidromic action potentials evoked from frontal cortex of urethane anesthetized rats were recorded extracellularly from nucleus locus coeruleus. The threshold current necessary to evoke antidromic action potentials varied as a result of infusion of adrenergic agonists and antagonists into frontal cortex within 50 micrometer of the stimulating electrode. Local infusion of the alpha-adrenergic agonist clonidine produced a marked decrease in terminal excitability, while the alpha-antagonist phentolamine produced an increase in terminal excitability and was shown to reverse the effect of the agonist. Infusion of the beta-adrenergic agonist isoproterenol was without effect, although the beta-antagonist propranolol resulted in a decrease in terminal excitability. Infusions of potassium increased excitability of locus coeruleus terminals. Terminal excitability was seen to vary inversely with the rate of spontaneous or high frequency stimulation-induced firing of locus coeruleus neurons. From these observations, it may be inferred that activation or blockade of alpha-adrenergic presynaptic receptors results in changes in polarization and/or conductance of the noradrenergic synaptic endings. These results are discussed with respect to phenomena associated with the possible presynaptic regulation of neurotransmitter release. PMID- 6271344 TI - The relationship between morphine analgesia and the activity of bulbo-spinal serotonergic system as studied by tolerance phenomenon. AB - The effect of various doses of acute morphine on both analgesia and 5 hydroxytryptamine (5-HT) synthesis in the brain and the spinal cord has been studied in rats rendered tolerant by chronic administration of the analgesic. In morphine-tolerant rats, the incorporation of tritiated-L-tryptophan (TRP) in the brain and the spinal cord was higher than in non-tolerant rats, but there was no significant difference in the synthesis rate of the newly formed 5-HT between the two groups. An acute dose of morphine (10 mg/kg) which induced a powerful analgesia and a large increase in 5-HT synthesis in non-tolerant rats, did not produce analgesia nor changes in 5-HT synthesis in tolerant rats. Higher acute doses of morphine which restored analgesia in tolerant rats, induced a discrete increase in [3H]TRP incorporation and a marked increase in 5-HT synthesis in the spinal cord of these animals. The same doses significantly increased [3H]TRP incorporation in the forebrain but did not modify 5-HT synthesis. These results show that tolerance to morphine is associated with a decrease in the effects of the drug on 5-HT synthesis in the spinal cord and the brain and tend further support to the hypothesis that an enhancement of 5-HT synthesis in the spinal cord, induced independently of modifications of the availability of TRP, is associated with the analgesic effect of morphine. PMID- 6271345 TI - Central nervous system myelin proteins and glycoproteins in vertebrates: a phylogenetic study. AB - CNS myelin was isolated by a conventional method from a wide range of vertebrate classes and analyzed by SDS-PAGE for proteins (Coomassie blue) and glycoproteins (concanavalin A (Con-A)-peroxidase). Mammalian, avian and reptilian myelin shared similar protein patterns (basic protein, BP; intermediate protein, DM-20; proteolipid protein, PLP; Wolfgram protein, W). Amphibians lacked DM-20 but were characterized by specific activities of 2',3'-cyclic nucleotide 3' phosphodiesterase (CNP) higher than those of the other classes examined. The Con A-binding profiles were similar in the high molecular weight (HMW) regions of the classes listed above, while the typical myelin proteins in the low molecular weight (LMW) regions were devoid of Con A-binding properties. In teleost myelin a putative BP band ran well ahead of rat small basic protein (SBP), whereas the region corresponding to rat PLP was covered by several closely spaced bands, most of which bound Con A. In elasmobranch myelin, apart from bands corresponding to BP, Con A-binding glycoproteins were detected migrating in the region of rat DM 20 and PLP as well as with mammalian PNS P0 protein. Cyclostomates yielded only very small amounts of material in the myelin preparation and displayed undifferentiated Coomassie blue- and Con A-binding in the HMW region, while typical LMW myelin proteins were absent. These results demonstrate that CNS myelin from bony and cartilaginous fishes is characterized by containing several major Con A-binding proteins of low molecular weight. This is in striking contrast to myelin from phylogenetically higher classes. PMID- 6271348 TI - Extracellular calcium and action potentials of soma and dendrites of hippocampal pyramidal cells. AB - Calcium activity ([Ca2+]0) and focal potentials were recorded from dendritic and cell body layers of CA1 and CA3 regions of slices of hippocampal tissue. Responses were evoked by focal stimulation of afferent fiber bundles. In order to evoke a detectable decrease of [Ca2+]0, stimulation had to be intense enough to cause postsynaptic discharge of action potentials; evoking EPSPs alone was insufficient. Responses of [Ca2+] were consistently greater in stratum pyramidale than in stratum radiatum. It is concluded that in hippocampal tissue activation of soma and proximal dendritic membranes is the most important contribution to the decrease of [Ca2+]0 in response to afferent stimulation. PMID- 6271347 TI - Neurons in the frontal cortex of the rat carry multiple opiate receptors. AB - Acute desensitization to the inhibitory action of iontophoretically applied opiate alkaloids and opioid peptides was used to investigate the possibility of multiple opiate receptors located on single neurons in the frontal cortex of rats. Such short term exposure resulted in adaptive processes which were similar to tolerance to and dependence on opiate agonists occurring after chronic treatment. Neurons desensitized to methionine-enkephalin (ME) or D-Ala2,D-Leu5 enkephalin (DADL) became subsensitive to morphine, whereas cells desensitized to morphine remain sensitive to the inhibitory action of the opioid peptides. This lack of cross-desensitization may suggest the existence of multiple opiate receptors on the same cell. PMID- 6271349 TI - An autoradiographic map of (3H)diprenorphine binding in rat brain: effects of social interaction. AB - (3H)Diprenorphine binding was analyzed autoradiographically in the brains of 33 day old rat pups. A photographic atlas of diprenorphine binding in the coronal plane is provided to highlight the dispersion of opioid receptor systems through the brain. To determine whether brain opioid release may be induced by social interactions, half the animals were sacrificed following a 30 min period of social interaction while the other half were sacrificed following 30 min of social isolation. Opioid binding was higher in isolate-tested animals than socially-tested ones, suggesting that social interaction may promote endogenous brain opioid release. PMID- 6271346 TI - Solubilization of the glycine receptor from rat spinal cord. AB - Glycine receptors, as detected by glycine-displaceable [3H]strychnine binding, were solubilized from a membrane fraction of rat spinal cord by the non-ionic detergent Triton X-100. The solubilized material retained its high affinity for [3H]strychnine and exhibited the typical pharmacological properties of the membrane-bound glycine receptor. On sucrose density gradients, the solubilized receptor had a sedimentation coefficient of 8.3 +/- 0.4 S. Gel exclusion chromatography on Sepharose 6 B in the presence of phosphatidylcholine gave a Stokes radius of 7.3 +/- 0.3 nm. PMID- 6271350 TI - [Study on the circulation of enterovirus 71 in the children population of Slovakia (author's transl)]. PMID- 6271351 TI - [Correlation between renal compensatory hypertrophy and urinary sodium excretion in the ACTH treated rat]. AB - In the rat, renal compensatory hypertrophy (RCH) is enhanced by the administration of ACTH (beta 1-24 corticotrophin) given at a dose of 18 micrograms/100 g. body weight/d. after uninephrectomy. In the rats drinking tap water, the enhancement of RCH by ACTH occurs without any significant change in urinary Na+ excretion. But, in the animals drinking a NaCl solution (9 g/l), ACTH determines a polyuria with hypernatriuria and a more striking increase of RCH positively correlated with the urinary Na+ excretion. PMID- 6271352 TI - [Effects of the injection of kainic acid into the medial hypothalamus on the secretion of corticolipotropic hormones in the rat]. AB - alpha-MSH secretion was significantly lowered in Rats after lesion of the mediobasal hypothalamus with microinjections of kainic acid. beta-Endorphin release after ether-stress was also reduced. In opposition, no significant change in basal and ether-stress ACTH secretion was observed. PMID- 6271353 TI - [Ontogenesis of the production of cyclic AMP mediated by histamine H2 receptors in glands isolated from gastric mucosa of the rat]. PMID- 6271354 TI - [Original hypothesis on the specificity of the binding of glycoprotein hormones to their receptors]. AB - From the data we have obtained in the last few years concerning the interaction of gonadotropins with their receptors in vitro we develop a coherent model of the mechanism responsible for the binding specificity of these hormones. In this model we postulate that there is structural separation between the sites responsible for the high affinity of this class of hormones for their receptors and the sites responsible for the specificity of binding of these hormones. The high affinity site would be common to all the glycoprotein hormones and would be mainly formed by the alpha subunit which is common to all these hormones. The specific sites would act by inhibiting the binding of each hormone to the receptors of the others. These sites would be borne mainly by the specific beta subunits. We have introduced the term "negative specificity" to describe this mechanism. PMID- 6271355 TI - Role of osteoblasts in hormonal control of bone resorption--a hypothesis. PMID- 6271356 TI - Calcium-mediated enhancement of the cyclic AMP response in cultured bone cells. AB - We have examined the influence of extracellular Ca2+ on cyclic AMP metabolism in an osteoblast-enriched population of bone cells isolated from the calvaria of rat fetuses. The cyclic AMP response to stimulators of cyclic AMP formation (PTH and PGE2), but not basal cyclic AMP levels, increased progressively as the extracellular Ca2+ concentration was raised from 0.2 to 4.0 mM. The response to changes in extracellular Ca2+ were rapid (within 3.5 min), and the level of responsivity that characterized each Ca2+ concentration persisted for at least 6 h when the Ca2+ concentration was kept constant. The effect of Ca2+ spanned the entire time course of PTH action, was not accompanied by altered excretion of cyclic AMP from the cells, and was evident at low as well as at high hormone concentrations. Ca2+ augmented the action of PTH in the presence as well as in the absence of cyclic AMP phosphodiesterase inhibitors, and failed to decrease cyclic AMP phosphodiesterase activity in the short term. Mn2+ and, to a smaller degree, Ba2+ substituted for Ca2+ in promoting the cyclic AMP response to PTH. Verapamil, an inhibitor of Ca2+ penetration, blunted the Ca2+-mediated increments in the cyclic AMP response, and the divalent cation ionophore A23187 enhanced these increments. These results indicate that Ca2+ and other cations are positive effectors of the stimulated cyclic AMP response in isolated bone cells. Accumulation into an as yet unknown cellular compartment may be required for the cation effect. The data are most consistent with enhancement of adenylate cyclase reactivity as the mode of cation action. PMID- 6271358 TI - Oligo(A)-stimulated Tetrahymena rDNA synthesis in vito. AB - The synthesis of Tetrahymena rDNA has been examined using purified DNA polymerase and partially purified preparations of homologous replication enzymes (fraction IV). DNA synthesis with purified DNA polymerase alone was less than that with fraction IV enzymes. This suggested that there were additional factors in fraction IV other than DNA polymerase which contributed to or enhanced rDNA synthesis in vitro. Neither hybridization of rDNA with Tetrahymena ribosomal RNA nor preincubation of rDNA with homologous or heterologous RNA polymerase served to stimulate in vitro synthesis by fraction IV enzymes. However, when rDNA was hybridized with oligoriboadenylate, DNA synthesis using fraction IV was stimulated approximately 4- to 4.5-fold over 150 min of incubation, relative to a similarly treated but unhybridized rDNA control. Using oligoriboadenylate hybridized EcoR1 and HindIII restriction fragments of rDNA to localize the synthesis most of the in vitro synthesis occurred with a 2.4 X 10(6) Mr fragment encompassing the centre of the rDNA molecule. The approach of hybridizing a synthetic homooligoribonucleotide primer to double-stranded DNA should prove to be of general applicability in designing similar template-primers in other systems for the purpose of isolating replication proteins. PMID- 6271357 TI - Decreased 1,25-dihydroxycholecalciferol and increased 25-hydroxy- and 24,25 dihydroxycholecalciferol in tissues of rats treated with thyroxine. AB - The effect of thyroxine on the metabolism of vitamin D was investigated in rats. Vitamin D depleted rats were repleted by injections of radiolabelled cholecalciferol or 25-hydroxycholecalciferol (25OHD3). After 3 weeks, a state of hyperthyroidism was induced by daily injections of L-thyroxine for 21 days. The lipid extracts of the Plasma and tissues were analyzed by successive Sephadex LH 20 and high pressure liquid chromatography. The concentrations of 25OHD3 and of 24,25-dihydroxycholecalciferol (24,25 (OH)2D3) were significantly higher and those of cholecalciferol and of 1,25-dihydroxycholecalciferol (1,25(OH)2D3) were significantly lower in the plasma and tissues of animals treated with thyroxine than in controls. The present study suggests that alterations in the metabolism of vitamin D may be involved in the disturbances of calcium metabolism observed in hyperthyroidism. PMID- 6271359 TI - Hydrolysis of triolein, cholesterol oleate, and 4-methylumbelliferyl stearate by acid and neutral ester hydrolases (lipases) from pigeon adipose tissue: effect of cAMP-dependent protein kinase. PMID- 6271360 TI - The incorporation of choline into disaturated phosphatidylcholine in the microsomal, lamellar body, and surfactant fractions of hamster lung tissue. AB - The specific activity of disaturated phosphatidylcholine in microsomes and lamellar bodies prepared from hamster lung tissue and in surfactant obtained by lung lavage was determined at various times following the intraperitoneal administration of [Me-3H]choline. The highest specific activity of disaturated phosphatidylcholine in the lung microsomes was attained 1 h after the administration of [3H]choline; thereafter, the specific activity declined. The specific activity of disaturated phosphatidylcholine in lamellar bodies increased steadily for 12 h after [3H]choline administration. The specific activity in lamellar bodies ater 12 h exceeded the maximum specific activity achieved in the microsomal fraction (p less than 0.005). The specific activity of the disaturated phosphatidylcholine in the alveolar lavage increased after an initial lag period of approximately 3 h, attaining the same specific activity as that of the lamellar bodies at the 12-h time point. The reported results are discussed in relation to the biosynthesis, storage, and secretion of the disaturated phosphatidylcholine associated with the lipoprotein, surfactant. PMID- 6271361 TI - Factors affecting the 16 alpha-hydroxylation of estrone 3-sulfate by guinea pig liver microsomes. AB - The estrone 3-sulfate 16 alpha-hydroxylase of guinea pig liver microsomes has been demonstrated to be sensitive to CO. A CO/O2 ratio of 0.64 caused 50% inhibition of activity. Since inhibition was also obtained in the presence of 2 diethylaminoethyl-2,2-diphenylvalerate . HCl it seems likely that the hydroxylase is a cytochrome P450 containing system. A fourfold increase in enzyme activity was brought about by 40 mM Mg2+ or Ca2+ while the same concentration of Mn2+ resulted in a twofold increase. Lesser increases were seen with Na+ or K+ and complete inhibition was obtained in the presence of Fe2+, Cu2+, or EDTA. When assayed in the presence of detergent concentrations sufficiently small to guard against cytochrome P450 destruction, it was found that Cutscum, Triton X-100, and Triton N-101 each caused greatest inhibition of enzyme activity. Lesser inhibition was apparent in the presence of Miranol H2M, cholate, or deoxycholate. The nonionic detergent, Brij 35, caused least inhibition of all and, when hepatic microsomes were treated higher concentrations of Brij 35, about 80% of protein and over 95% cytochrome P450 were to be found in the 100 000 X g supernatant. Microsomal activity was more stable when stored at -20 degrees C in buffer containing glycerol, EDTA, and dithiothreitol than in buffer alone. Under best conditions only 10% of the hydroxylase activity was lost in one week. PMID- 6271362 TI - The phosphatidyl-myo-inositol-4,5-biphosphate phosphatase from Crithidia fasciculata. AB - The phosphatase which specifically removes one phosphate group from phosphatidyl myo-inositol 4,5-bisphosphate was purified up to 6000-fold from the cytosol of the protozoan Crithidia fasciculata. Lipoproteins which interfere with the purification were precipitated by reducing the pH to 4.5. The enzyme was isolated from the supernatant by ammonium sulfate fractionation, gel filtration (Sepharose CL-6B), ion-exchange chromatography (DEAE-Sepharose CL-6B), and hydrophobic chromatography on detergent-saturated phenyl-Sepharose CL-4B. The preparations had specific activities of 44-110 mumol . min-1 . mg protein-1 with phosphatidyl myo-inositol 4,5-bisphosphate, but were inactive with a variety of lipid and nonlipid phosphate esters. The enzyme was stable in the presence of salt and exhibited a relative mass of 117 000. It formed larger aggregates in the absence of salt and was dissociated into monomers of relative mass 57 000 by sodium dodecyl sulfate. Addition of Triton X-100 to the assay mixture reduced the dependence upon moderation of the charge of the substrate by cetyltrimethylammonium bromide. In the presence of both detergents the Mg2+ dependence of the enzyme was reduced (Km for Mg2+ = 40 microM) while the "apparent" Km for the substrate was unchanged at 240 microM. Substrate precipitation at higher Mg2+ concentrations was eliminated. PMID- 6271363 TI - Cell-free ammonia-oxidizing system of Nitrosomonas europaea: general conditions and properties. AB - Conditions essential for the preparation of active ammonia-oxidizing extracts of Nitrosomonas europaea were studied. The extracts were unstable during storage and required specific assay conditions for ammonia oxidation. Bovine serum albumin, spermine, or MgCl2 was required for ammonia oxidation and the concentration of phosphate determined the relative effectiveness of each activator, i.e., albumin being most effective in 0.1 M phosphate and spermine or MgCl2 at lower phosphate concentrations. Kinetic experiments showed a partial reduction of cytochrome c preceding the initiation of oxygen consumption due to ammonia oxidation. PMID- 6271364 TI - A partial resolution and reconstitution of the ammonia-oxidizing system of Nitrosomonas europaea: role of cytochrome c554. AB - The cell-free ammonia-oxidizing system of Nitrosomonas europaea was resolved into three major fractions: a membrane fraction containing cytochrome a1 and c-type cytochromes, a fraction with hydroxylamine-cytochrome c reductase and a cytochrome c fraction. The ammonia-oxidizing activity was reconstituted by the combination of these three fractions. The activity was more consistently reconstituted by adding Nitrosomonas cytochrome c554 to the membrane fraction. The hydroxylamine-cytochrome c reductase activity of the membrane fraction increased with the addition of cytochrome c554, but the oxidation of hydroxylamine to nitrite required a further addition of cytochrome c552. The ammonia oxidation by the membrane plus cytochrome c554 was affected by the concentration of phosphate and the addition of bovine serum albumin, spermine, or MgCl2. PMID- 6271365 TI - Effect of delta 9-tetrahydrocannabinol on cyclic nucleotides in synchronously dividing Tetrahymena. AB - Cyclic nucleotide levels were determined in division-synchronized Tetrahymena and the effect of delta 9-tetrahydrocannabinol (THC) on the cyclic nucleotide levels was studied. In non-drug-treated division-synchronized cells, there was no statistically significant variation in the level of cAMP and cGMP during the G2 period, preceding the first division. During the free running cell cycle (the interval of time between the first and second synchronous division) the twofold increase in the level of cAMP was statistically significant; however the variation in the level of cGMP was not statistically significant. THC caused a lowering of cAMP and cGMP levels throughout the 4-experimental treatment. The suppression of cAMP and cGMP levels altered the cyclic nucleotide pattern of the cell cycle. The cAMP pattern was changed particularly in the G2 period preceding the first synchronous division, and immediately after division during the free running cell cycle. THC treatment caused division delays of approximately 8-15 min in the onset of the first and second synchronous division. However, the duration of the free running cell cycle (110-120 min) was unchanged. The suppression of cyclic nucleotide levels resulting from THC treatment is discussed in relation to delays in the division schedule. PMID- 6271367 TI - The specific capsular polysaccharide of Streptococcus pneumoniae type 9V. AB - The specific capsular polysaccharide produced by Streptococcus pneumoniae type 9V (American type 68) is composed of D-glucuronic acid (1 part), D-galactose (1 part), 2-acetamido-2-deoxy-D-mannose (1 part), D-glucose (2 parts), and O-acetyl (1.6 parts). Methylation, periodate oxidation, optical rotation, and nuclear magnetic resonance studies, and partial hydrolysis showed that the polysaccharide is an unbranched high molecular weight linear polymer of a partially O-acetylated pentasaccharide repeating unit having the structure indicated below. (Formula: see text). PMID- 6271366 TI - Pulmonary phosphatidic acid phosphohydrolase: further studies on the activities in rat lung responsible for the hydrolysis of membrane-bound and aqueously dispersed phosphatidate. AB - Rat lung cytosol and microsomal fractions both contain phosphohydrolase activity towards membrane-bound phosphatidic acid (PAmb) and aqueously dispersed phosphatidic acid (PAaq) which cannot be explained through contamination with the other fraction. The phosphohydrolase activities with PAaq demonstrated Km and Vmax values which were more than an order of magnitude greater than those observed with PAmb and with vesicles prepared from the lipids extracted from [32P]PA-labelled microsomes. The PAaq-dependent activities in both fractions were stimulated by preparing mixed liposomes with phosphatidylcholine. The PAmb dependent activities in rat lung microsomes and cytosol were markedly stimulated by high concentrations of Triton X-100 and Nonidet P-40. The PAmb- and PAaq dependent activities in the microsomes were stimulated by deoxycholate. Although no difference was observed in the inhibition profiles of the PAmb- and PAaq dependent activities of the cytosol in the presence of various mercurials, the PAmb-dependent activity in the microsomes was somewhat more susceptible than the PAaq-dependent activity. The PAmb-dependent activities in both fractions were more susceptible to inhibition by iodoacetamide. These results support the view that separate rat lung enzymes were involved in the hydrolysis of PAmb and PAaq. The relative abilities of rat lung cytosol and microsomes to hydrolyse PA endogenously generated on the microsomes were compared using relative concentrations of cytosol corresponding to the levels in intact rat lung. During the initial period (5-10 min) the cytosol phosphohydrolase activity was more effective than the microsomal activity. At later stages (10-20 min), the rates were comparable. PMID- 6271368 TI - Effects of a methyl-deficient diet on rat liver phosphatidylcholine biosynthesis. AB - To produce a severe choline-methionine deficiency, a synthetic L-amino acid diet, free of choline, methionine, vitamin B12, and folic acid and supplemented with guanidoacetic acid, a methyl group acceptor, was fed to female rats for 2 weeks. The in vitro activity of liver microsomal phosphatidylethanolamine methyltransferase was stimulated twofold when compared with basal diet controls. The activity of choline phosphotransferase was depressed by 86%; thus, the contribution of the methyltransferase in the overall synthesis of phosphatidylcholine apparently increased. However, measurement of the in vivo methylation of phosphatidylethanolamine by incorporation of [1,2-14C]ethanolamine into phosphatidylcholine indicates that the methylation pathway is markedly depressed in methyl deficiency. Hepatic concentrations of the methyltransferase substrate, S-adenosylmethionine, and the inhibitory metabolite, S adenosylhomocysteine, were significantly altered such that an unfavorable environment for methylation was present in the deficient animal. The ratio of substrate to inhibitor was depressed from 5.2:1 in the controls to 1.7:1 in the livers of methyl-depleted rats. Control of transmethylation in accordance with the availability of substrates, phosphatidylethanolamine, or S adenosylmethionine, and the level of S-adenosylhomocysteine is discussed. PMID- 6271369 TI - The relationship between variations in levels of serotonin acetyltransferase activity and cGMP content in cultured chick pineal glands. PMID- 6271370 TI - Brown adipose tissue mitochondria of cold-acclimated rats: change in characteristics of purine nucleotide control of the proton electrochemical gradient. PMID- 6271371 TI - Metabolism of cytoplasmic triacylglycerol in cultured aortic smooth muscle cells. AB - A turnover of cytoplasmic triacylglycerol was studied in cultured rat, rabbit, and bovine aortic smooth muscle cells. Cytoplasmic triacylglycerol was labeled with [3H]glycerol in the presence of oleic acid in the medium and its loss from the cell was studied in the presence of carrier glycerol. Multiple additions of Isuprel or dibutyryl cyclic AMP during the chase period did not enhance the loss of labeled triacylglycerol. The rate of hydrolysis of cellular triacylglycerol was unchanged in the absence or in the presence of 100 microM chloroquine. Modulation of cellular cholesterol content by addition of low density lipoprotein or high density apolipoprotein--sphingomyelin liposomes did not affect the residence time of the cellular triacylglycerol. We conclude that cytoplasmic triacylglycerol in cultured aortic smooth muscle cells is metabolized by an extralysosomal enzyme which is neither catecholamine responsive nor affected by modulation of cellular cholesterol. PMID- 6271372 TI - Synaptosomal Na+,K+-ATPase as a membrane probe in studying the in vivo action of morphine. AB - The activity of membrane-bound Na+,K+-ATPase was used as a metabolic probe to study the effects of morphine in vivo in rat brain synaptosomes. Arrhenius plots were generated to study an induced perturbation within the membrane. In acute studies 0.5-h postmorphine, the drug was without effect on the basal activity of the enzyme. With dopamine-stimulated Na+,K+-ATPase morphine decreased the apparent transition temperature and specific activity of the enzyme while there was a slight stimulation in its activation energy. An increase in these parameters was observed in samples taken from animals withdrawn from the drug for 48 h. These results strongly suggest the possible involvement of the membrane phospholipids as transducer which mediates the observed biphasic effect of the drug on enzyme activity. PMID- 6271373 TI - Antagonism of presynaptic adenosine receptors by theophylline 9-beta-D-riboside and 8-phenyltheophylline. AB - Theophylline 9-beta-D-riboside and 8-phenyltheophylline were evaluated as presynaptic adenosine receptor antagonists in the rat vas deferens in vitro. Stimulation of presynaptic adenosine receptors, which results in an inhibition of the twitch response to electrical field stimulation, was achieved with 2 chloroadenosine, an adenosine analogue that appears not to be a substrate for the adenosine transport system. The presynaptic inhibitory action of 2 chloroadenosine was antagonized by theophylline (10 and 100 microM) and by 8 phenyltheophylline (10 microM) but not by theophylline 9-beta-D-riboside (100 microM). It is concluded that the addition of a ribose moiety to theophylline does not enhance the antagonist potency of the molecule but actually renders the compound inactive. However, 8-phenyltheophylline is approximately three times more potent than theophylline at presynaptic adenosine receptors. PMID- 6271374 TI - Effects of tonin on the response to norepinephrine by the aortic strip of the hypertensive rat. AB - The response to norepinephrine (NE) of arterial smooth muscle from two types of experimental hypertensive rats was investigated. Aortic strips from one-kidney, one-clip hypertensive animals were less responsive to NE than those from their normotensive controls but strips from one-kidney, one-clip hypertensive animals showed no difference from their corresponding controls. The contractility in response to NE was the same in all groups. These results suggest that the mechanisms responsible for lesser reactivity in the one-kidney hypertensive group are not a consequence of elevated blood pressure itself but may be related to changes in the intrinsic sensitivity of aortic smooth muscle. Tonin potentiated the contraction induced by NE in aortic strips from hypertensive and normotensive rats. This effect was more pronounced in the one-kidney, one-clip hypertensive animals, so that although the aortic smooth muscle from these animals is less reactive to NE, the decreased reactivity can be more than compensated by the presence of tonin. The mechanism of potentiation is not yet clear but the fact that Saralasin did not inhibit it suggests that angiotensin II is not generated in situ. PMID- 6271375 TI - Detection from rat pituitary of beta-lipotropin and materials containing opiatelike activity by combined enzymatic radioreceptor assay. AB - Tonin, a proteolytic enzyme isolated from rat submaxillary gland, was allowed to react upon ovine beta-lipotropin (beta-LPH) at 37 degrees C at a variety of pH values and for different lengths of time. Opiatelike activity generated by the reaction was assessed using a radioreceptor assay for beta-endorphin with rat brain homogenate. [3H]naloxone, and beta-endorphin as receptors, tracer, and hormone standard, respectively. Cleavage of beta-LPH with tonin produced a 10 fold increase in opiatelike activity as compared with beta-LPH alone. Digestion of beta-LPH with other enzymes such as renin, cathepsin D, trypsin, and chymotrypsin produced much less opiatelike activity. beta-Endorphin and methionine-enkephalin were not cleaved by tonin. Using this new assay, we were able to detect beta-LPH and materials containing opiatelike activity from rat pituitary extracts after gel chromatography. It is more specific and more sensitive than trypsin digest. PMID- 6271376 TI - Insulin and glucagon in rats with islet cell tumors induced by small doses of streptozotocin. AB - Plasma insulin and glucagon responses to oral glucose loading were examined in rats with islet cell tumors induced by a single intravenous injection of streptozotocin (30 or 40 mg/kg body weight). Twenty-four macroscopic and six microscopic tumors occurred in 21 rats. In 15 of 21 tumor-bearing rats, there was exaggerated insulin release in response to oral glucose. Plasma glucose levels did not rise with the oral glucose load and were comparable to those seen in normal animals. Hence these rats are described as having "responsive tumors." In six rats with "nonresponsive tumors" there was no insulin response and the plasma glucose levels rose. No significant differences in plasma levels were observed between the two groups. Nonresponsive tumors as well as responsive tumors contained a significant amount of extractable insulin (17.68 +/- 8.60 and 35.07 +/- 10.05 mg/g wet weight, respectively) and detectable amounts of immunoreactive glucagon (1.47 +/- 0.61 and 2.24 +/- 0.67 micrograms/g wet weight, respectively). These results suggest that a small dose of streptozotocin produces two types of islet cell tumors. One is insulin producing and insulin secreting whereas the other is insulin producing but not insulin secreting. PMID- 6271377 TI - Pressor effect of tonin in anephric animals. AB - Tonin was injected intravenously to normal rats without effect on blood pressure. Twenty-four hours after bilateral nephrectomy, tonin produced a dose-dependent pressor effect in rats which was abolished by the angiotensin antagonist [Sar1 Ala8]-angiotensin II. Vascular response to angiotensin II was slightly increased after nephrectomy. Plasma angiotensin II increased significantly after injection of tonin and disappeared biexponentially with a half-life of less than 1 min for the fast component and 9 min for the slow component. The change in plasma angiotensin II correlated with the elevation in mean blood pressure. No difference in inhibitory power of plasma on tonin activity could be shown between intact and nephrectomized rats. In vitro, the initial velocity of generation of angiotensin II by tonin acting on plasma increased after addition of semipurified rat renin substrate and was significantly greater in plasma of nephrectomized rats. In nephrectomized rabbits, but not in intact ones, a dose-dependent pressor effect was produced by tonin. These data demonstrate the in vivo production of angiotensin II by tonin in an animal model with elevated substrate levels. Together with the in vitro data, these results suggest a role for substrate concentration in the expression of tonin enzymatic activity in vivo. PMID- 6271378 TI - Inhibition of the n-formylmethionyl-leucyl-phenylalanine induced respiratory burst in human neutrophils by adrenergic agonists and prostaglandins of the E series. AB - Exogenous prostaglandins E1 and E2 and L-isoproterenol potently inhibited the production of superoxide anions by human neutrophils activated in vitro by n formylmethionyl-leucyl-phenylalanine (FMLP). An estimated ID50 of 50 nM was found for all three agents while L-epinephrine and prostaglandin F2 alpha were 10 and 100 fold, respectively, less active. Inhibition occurred whether these agents were added before, together with, or after the addition of the tripeptide to cell suspensions. Cells treated with dibutyryl adenosine 3',5'-monophosphate also expressed reduced rates of superoxide synthesis thus suggesting that the hormonal inhibitors acted indirectly by stimulating membrane bound adenylate cyclase. PMID- 6271379 TI - 2-phosphoglycerate phosphatase and serine biosynthesis in Veillonella alcalescens. AB - The constituent enzymes for the phosphorylated and nonphosphorylated serine biosynthetic pathways in Veillonella alcalescens were identified and included phosphoserine phosphatase, 3-phosphoglycerate dehydrogenase, glycerate dehydrogenase, phosphoserine aminotransferase, and serine-pyruvate aminotransferase. Cell extracts of the organism were also found to cause the specific dephosphorylation of 2-phosphoglycerate. The phosphatase was purified 39 fold by manganese chloride precipitation, ammonium sulfate precipitation, and DEAE-cellulose chromatography. Sephadex G-200 gel filtration data established an apparent molecular weight of 50000 for the enzyme. The 2-phosphoglycerate phosphatase had a pH optimum of 5.5 and was distinct from phosphoglyceromutase. Assays conducted with the purified enzyme on a number of other phosphorylated intermediates indicated that the phosphatase was most specific for 2 phosphoglycerate. Glucerate, hydroxypyruvate, and serine inhibited the enzyme, whereas succinate stimulated activity. Veillonella 2-phosphoglycerate phosphatase is the first such enzyme to be described in a prokaryote and is probably involved in glycerate generation for the nonphosphorylated serine biosynthetic pathway. PMID- 6271380 TI - Experimental porphyric neuropathy: a preliminary report. AB - An experimental model for the study of porphyric neuropathy is presented. Injection of either tetraphenyl-porphinesulfonate (TPPS), hematoporphyrin derivative (HpD), or delta-amino-levulinic acid (ALA) into mice resulted in markedly decreased motor nerve conduction velocity (MNCV). THe MNCV returned to normal within one week following the injection of large doses of ALA, and within three weeks following the injection of close to lethal doses of HpD, but there was no recovery of nerve function within 50 days following injection of substantially smaller doses of TPPS. Ultrastructural examination of motor nerves at various times following TPPS injection revealed the gradual development of structural abnormalities. Ultrastructural examination of the same nerves after a single dose of either ALA or HpD failed to demonstrate any abnormalities. The present observations call for precaution as to the use of TPPS as photosensitizer in human cancer treatment. PMID- 6271381 TI - Evaluation of alternating chemotherapy and sites and extent of disease in extensive small cell lung cancer. AB - Sixty-six patients were entered into a prospective, randomized clinical trial evaluating the use of alternating noncross-resistant chemotherapy in patients with extensive small cell carcinoma of the lung. Sixty-five were evaluable. One regimen utilized cyclophosphamide, VP-16, vincristine, cisplatin alternating with doxorubicin (Adriamycin) and DTIC (CVVP-AD). The second regimen utilized doxorubicin, VP-16, vincristine, and cisplatin alternating with cyclophosphamide and DTIC (AVVP-CD). There was no statistically significant difference between the two chemotherapeutic programs in terms of regression rate, time to progression, or survival. Overall regression rate for CVVP/AD was 91% (29/32) including five complete regressions (CRs). For AVVP-CD, the total regression rate was 82% (27/33) including nine CRs. Combined, the overall regression rate was 86% with a CR rate of 22%. Time to progression for CVVP-AD and AVVP-CD was 28 and 26 weeks, respectively. The median survival time of CVVP-AD and AVVP-CD regimens was 40 and 42 weeks, respectively. Prognostic variables significantly correlated with survival were performance score and extensive liver metastases at diagnosis. Correlations between initial sites of disease led to the observation that patients with no central nervous system (CNS) metastases at diagnosis were more likely to have more extensive liver and lung involvement. Further analysis revealed the lung to be the most common site of first progression (46%) and liver second (28%). Patients with extensive involvement of the liver or lung progressed sooner in these sites than those with a lesser tumor extent. At some point in the study, 40% of the patients experienced CNS metastases. The efficacy of these two alternating regimens is comparable to most current regimens reported in extensive SMCLC. Whether cyclophosphamide or doxorubicin is used first seemed to make little difference. The alternate noncross-resistant regimen was rarely effective in producing tumor regression following initial progression. PMID- 6271383 TI - Laryngeal carcinoid tumor: light and electron microscopic studies. AB - A case of laryngeal carcinoid in a Japanese 76-year-old man is reported. This neoplasm is extremely rare in the larynx and the case is the fourth so far reported in the world medical literature. Light and electron microscopic studies revealed argyrophilia and the presence of neurosecretory granules that had not been demonstrated in the previous cases. A review of the medical literature showed that all laryngeal carcinoids were pathologically malignant. The relationship of carcinoid tumor to oat cell carcinoma of the larynx is discussed, and the suggestion is made that a similar relationship exists between them as between carcinoid tumors and oat cell carcinomas of the lung. PMID- 6271382 TI - Herpes simplex infection in acute myelogenous leukemia and other hematologic malignancies: a prospective study. AB - To better define the frequency and clinical characteristics of herpes simplex virus (HSV) infection in adult patients with acute myelogenous leukemia (AML), the authors prospectively studied 29 patients undergoing remission induction chemotherapy with twice weekly throat wash cultures for an average of 25.3 days. Ten seropositive patients (34.5%) shed HSV at least once. Eight patients were asymptomatic. Two episodes of herpes labialis were severe and persistent, but no visceral dissemination was observed. Reactivation of HSV infections in AML patients presumably with marked immunosuppression occurs, but less frequently and more benignly than has been suggested. Daunomycin and cytosine arabinoside, which can inhibit HSV replication, may have accounted for this lower frequency of reactivation. PMID- 6271384 TI - Small cell lung cancer: analysis of treatment factors contributing to prolonged survival. AB - Seventy-one consecutive patients with small cell lung cancer (SCLC) were treated in the Radiation Oncology Branch of the National Cancer Institute using six different radiation regimens in combination with cyclophosphamide, vincristine and doxorubicin chemotherapy. Patients treated with concurrent chemotherapy irradiation (CT-RT) experienced better local tumor control than patients treated with sequential CT-RT. Maximum survival with minimum toxicity occurred in the group given a three-week course of concurrent CT-RT. Although concurrent therapy appeared more toxic than sequential therapy, it also appeared to result in more effective tumor control. Precise details of the timing of CT and RT represent important variables in study design of combined modality therapy for SCLC. Carefully controlled clinical trials should be undertaken to define the optimal timing and sequencing of CT-RT, as well as the optimal dose of RT. PMID- 6271385 TI - Doxorubicin-hexamethylmelamine therapy of small cell carcinoma of the lung. AB - Eighteen patients with small cell carcinoma of the lung treated with doxorubicin hydrochloride and hexamethylmelamine are presented. Fifteen of these patients had extensive disease at presentation. Four patients in this group died after one or fewer courses of chemotherapy. The median duration of survival for the entire group of patients is 15 months. Six patients are alive from 18 to 56 months without evidence of disease. Drug toxicity was minimal and well tolerated, which permitted this regimen to be given in an outpatient setting. PMID- 6271386 TI - Advanced ovarian carcinoma: a prospective and randomized clinical trial of cyclophosphamide versus combination cytotoxic chemotherapy (Hexa-CAF). AB - Fifty-seven evaluable patients with advanced ovarian carcinoma were randomized to receive either a combination of hexamethylmelamine, cyclophosphamide, methotrexate and 5-fluorouracil (Hexa-CAF) or high-dose cyclophosphamide alone given intravenously intermittently. Objective responses were seen in 62% of patients receiving cyclophosphamide alone, and 36% of patients in the Hexa-CAF regimen, this difference being statistically significant (P less than 0.05). The median duration of objective response (10 months vs. 9 months) and the median survival (11 months vs. 10 months) were greater in the cyclophosphamide group, but these differences were not statistically significant. It is concluded that there is no therapeutic advantage for the Hexa-CAF protocol over the alkylating agent used alone. PMID- 6271387 TI - Primary radiation treatment of colloid carcinoma of the breast: a case report. AB - A case report is presented of a patient with a colloid carcinoma of the breast, stage T3 N0 M0, treated definitively with external and interstitial radiation. Three-year follow-up has shown no local recurrence, with complete regression and no development of regional or distant metastases. The prognostic significance of colloid carcinoma and the use of radiation in its treatment are discussed. PMID- 6271388 TI - Glycogen-rich clear cell carcinoma of the breast: a light and electron microscopic study. AB - A glycogen-rich clear cell carcinoma arose in the breast of a 49-year-old woman. Light microscopic examination of the neoplasm revealed both intraductal papillary growth and stromal invasion. Electron microscopic examination demonstrated neoplastic cells that contained massive quantities of nonmembrane-bound particulate glycogen and that formed numerous acini. Apically, these cells formed microvilli; laterally they formed tight junctions and desmosomes. Morphologic features of this neoplasm are similar to those of the fetal breast and to some other clear cell carcinomas arising elsewhere in the body. PMID- 6271389 TI - Glycosaminoglycans in human lung cancer. AB - The quantitative changes of glycosaminoglycans in tumor tissue of human lung cancers (2 squamous cell carcinomas, 4 adenocarcinomas and 5 small cell carcinomas) were studied. The total amount of glycosaminoglycans in human lung cancer tissues increased 1.4 to 4 times in comparison with that in normal lung tissues. The increase in tissue content of glycosaminoglycans was accompanied by an increase in the chondroitin sulfate level in every histologic type of lung cancer, as well as by a marked increase in hyaluronic acid level in squamous cell carcinomas, and a moderate increase in its level in small cell carcinomas. The concentrations of dermatan sulfate and heparan sulfate in lung cancer tissues did not show any significant changes compared with those in normal lung tissues. The increase in total amount and changes in the composition of glycosaminoglycans in human lung cancer tissue were closely related to the histologic type of the tumor. PMID- 6271390 TI - Ectopic hormone production by endocrine tumors: localization of hormones at the cellular level by immunocytochemistry. AB - Clinical and laboratory data, histologic, electron microscopic and immunocytochemical findings of the tumors of eight patients suffering from Cushing's syndrome and of one patient with hypercalcemia are described. The unlabeled antibody enzyme method was used for the detection of insulin, glucagon, somatostatin, pancreatic polypeptide, corticotropin, beta-lipotropin, calcitonin, parathyroid hormone, and gastrin. Ectopic Cushing's syndrome was caused by pancreatic endocrine tumors, medullary thyroid carcinoma, a bronchial, a gastric and a thymic carcinoid, and a carcinoid of the mediastinum. Hypercalcemia in one patient was related to a pancreatic endocrine tumor. After surgery the clinical symptoms disappeared in two patients, but persisted or relapsed in five patients. ACTH-immunoreactivity could be demonstrated in six of eight tumors; calcitonin immunoreactivity was found in the tumor of the patient suffering from hypercalcemia. ACTH-immunoreactivity could be localized to secretory granules by immunoelectron microscopy, and the presence of ACTH and beta-LPH in the same tumor cells could be shown in one pancreatic tumor. A combination of production of orthotopic and ectopic hormones was found in one, and secretion of two ectopic hormones was detected in another pancreatic endocrine tumor. PMID- 6271391 TI - The diagnostic dilemma of the "small round cell neoplasm": catecholamine fluorescence and tissue culture morphology as markers for neuroblastoma. AB - Small round cell neoplasms, which include neuroblastoma, Ewing's sarcoma, embryonal rhabdomyosarcoma, oat cell carcinoma, and lymphoma, are often confused with one another histologically. The advent of successful but distinct therapeutic approaches for different neoplasms has increased the need for precise diagnosis. The use of techniques ancillary to routine histologic or ultrastructural analysis allows better definition of the specific tumor type. Four cases of small cell tumor are described in which the initial working diagnosis was incorrect. The application of two diagnostic procedures for neuroblastoma was of great value to clarifying the proper diagnosis. These tests were a rapid fluorescence assay for intracellular catecholamines and a tissue culture assay for neurite outgrowth. Both of these methods are highly sensitive for neuroblastoma and distinguish neuroblastoma from other small round cell neoplasms. Their use confirmed this unsuspected diagnosis in three cases and excluded neuroblastoma in the fourth case. PMID- 6271392 TI - Induction of DNA repair synthesis in isolated rat hepatocytes by 5-diazouracil and other DNA damaging compounds. PMID- 6271393 TI - The effect of N-nitroso-2-methoxy-2,6-dimethylmorpholine on endocrine and exocrine pancreas of Syrian hamsters. AB - N-Nitroso-2-methoxy-2,6-dimethylmorpholine (MeNDMM), a cyclic derivative of the proposed proximate pancreatic carcinogen N-nitroso(2-hydroxypropyl) (2 oxopropyl)amine (HPOP), is shown to have an almost selective cytotoxic effect on pancreatic beta-cells when a single high dose is given to Syrian hamsters. Hence in this aspect its effect is comparable to that of streptozotocin, which has a glucose moiety similar to the MeNDMM structure. However, contrary to the effect of streptozotocin, low single (subdiabetogenic) doses of MeNDMM led to the development of pancreatic ductular and mixed ductular-insular neoplasms; only 1 animal also had islet cell adenoma. It therefore seems that MeNDMM possesses an affinity for both endocrine and exocrine pancreatic tissue. Other target tissues of MeNDMM were the forestomach, intra- and extrahepatic bile ducts, liver, kidneys and vagina. The tumors of these organs appeared in various incidences, partially in relation to dose and/or survival time. The possible mechanisms of the MeNDMM effect upon the endocrine and the exocrine pancreas is discussed. PMID- 6271394 TI - Removal of benzo[a]pyrene from cells by various components of medium. AB - Benzo[a]pyrene is removed from cells in culture by various additions to the medium. During post-treatment incubation, WI-38 fibroblasts were incubated with a low density, very low density and high density lipoproteins, delipidated or complete serum or plasma, or serum albumin. The time course of removal was followed. Increasing concentrations of lipoproteins resulted in increasing percentages of removal of benzo[a]pyrene from cell membranes. The most efficient addition was 10% complete human plasma. These results indicate that benzo[a]pyrene remains at or close to the plasma membrane for at least several hours and readily redistributes to medium components. PMID- 6271395 TI - Purine pathway enzyme abnormalities in acute lymphoblastic leukemia. AB - The status of three purine pathway enzymes, adenosine deaminase, 5'-nucleotidase, and purine nucleoside phosphorylase, was evaluated in the leukemic cells of patients with acute lymphoblastic leukemia and correlated with routine immunological cell surface markers. A distinct pattern of enzyme activity was noted in T-lymphoblasts which have significantly higher adenosine deaminase activity (p less than 0.02) and lower 5'-nucleotidase (p less than 0.001) and purine nucleoside phosphorylase (p less than 0.01) activities than do non-T, non B lymphoblasts. This enzyme pattern is similar to that observed in normal human thymocytes but is not shared by the mature, normal T-lymphocytes of peripheral blood, suggesting that it may reflect the differentiation status of malignant T lymphoblasts. These findings, which confirm the biochemical heterogeneity of acute lymphoblastic leukemia, may provide an avenue for selective chemotherapy of this disease. PMID- 6271396 TI - Cytochemical markers of differentiation in acute leukemia. PMID- 6271397 TI - Evaluation of an intermittent schedule of mitolactol in advanced non-small cell lung cancer. PMID- 6271398 TI - Does fluorescent analogue of propranolol, 9-amino-acridine-propranolol, binding sites really show the beta-adrenoreceptors? PMID- 6271399 TI - Inhibition and transgalactosylation studies on yeast alpha-galactosidase. PMID- 6271400 TI - Experimental studies on differentiation of cells originated from human neural crest tumors in vitro and in vivo. PMID- 6271401 TI - Effects of stimulators and inhibitors on arachidonic acid metabolism in hepatoma. PMID- 6271402 TI - Enhancement of DNA synthesis by biliverdin in a non-transformed liver cell strain. PMID- 6271403 TI - Epidermal growth factor stimulates DNA-synthesis of astrocytes in primary cerebellar cultures. AB - The capability of epidermal growth factor (EGF) to stimulate DNA-synthesis in neural cells was investigated in primary cultures of early postnatal mouse cerebellum. At concentrations of 10(-8)M, EGF stimulates DNA synthesis in astrocytes, which were identified immunocytologically by the cell type-specific marker, glial fibrillary acidic (GFA) protein. Astrocytes express cell-surface receptors for EGF as can be shown by binding of [125 I]-labeled EGF to live monolayer cultures. In the presence of 10% horse serum, EGF stimulates DNA synthesis by a factor of about two-fold. Stimulation by EGF over control values is approximately 4-fold in the presence of 1% serum and 6-to 10-fold in the absence of serum. Absolute numbers of astrocytes are increased after more prolonged action of EGF. DNA-synthesis in neurons or oligodendroglia is not significantly stimulated by EGF. EGF enhances cell survival of serum-deprived cerebellar cultures. Fibroblast growth factor does not increase DNA-synthesis in astrocytes under the conditions used in this study. PMID- 6271404 TI - Localization of sialic acid-containing hormones in GTH cells and ACTH cells of the rat anterior pituitary. AB - The localization of sialic acid-containing substances in the rat anterior pituitary gland has been studied by light and electron microscopy, using a peroxidase-labeled lectin (limulus polyphemus agglutinin: LPA) which binds specifically to sialic acid residues. LPA stains two types of anterior pituitary cells: (1) round or ovoid cells which are also positively stained with anti-hCG (GTH cell), and (2) small, stellate cells which are unstained with anti-hCG (ACTH cell). All of the LPA-positive cells can be distinguished from TSH cells which are identified by the use of anti-hTSHbeta. On ultrathin sections directly stained with LPA using the postembedding method, the reaction is confined to the secretory granules in GTH cells, and ACTH cells. Of two types of secretory granules in GTH cells, the larger one is intensely stained, whereas the smaller type shows only weak staining with LPA. Since follicle-stimulating hormone (FSH) is known to have high sialic acid contents, the results suggest possible detection of FSH with a technique other than immunohistochemistry. Furthermore, if the sialic acid-containing substances in GTH cells represents FSH, then these results support the hypothesis that LH cells and FSH cells are one cell type. PMID- 6271405 TI - Role of histamine and cyclic nucleotides in implantation in the rabbit. AB - The effect of histamine on cAMP and cGMP levels in day 6 (144 h post coitum) rabbit blastocysts was determined. Histamine at 200 microM and 1000 microM concentrations stimulated the increased formation of cAMP in vitro, whereas stimulation of cGMP occurred only in the presence of 1000 microM histamine. Furthermore, intrauterine injection of RMI-12330 A (50 microgram or 500 microgram/uterine horn), an inhibitor of adenylyl cyclases, on day 5 of pregnancy interrupted embryo development and implantation of the embryo. The drug was also effective in reducing the cAMP level in the endometrial cells in vitro. A relationship between histamine and cyclic nucleotide changes in embryo development and implantation is suggested. PMID- 6271406 TI - Effects of butylated hydroxyanisole on the aryl hydrocarbon hydroxylase of rats and mice. PMID- 6271407 TI - Formation of denatured erythrocytes by exposure to a superoxide radical generating system of xanthine oxidase. PMID- 6271408 TI - [Snakes endemic to plantations of the Ivory Coast]. AB - Authors describe the taxonomic composition of snake population in some plantations of Ivory Coast. In these areas, hazards of snake bites has been studied. They show that in extensive farming ecological conditions are strongly changed inducing modifications into the snake populations. In rustic farming area ecological conditions do not change with those of the brush or village surrounding. Snake bites in some extensive farming are ten times more frequent than in village or rustic plantations around them. This is linked to the snakes and farmers densities which are different in each plantation. Lethal effect is weak as in envenomations declared during any other activities. PMID- 6271409 TI - Reflux discharges recorded from rat submandibular ganglion cells in vivo. PMID- 6271410 TI - Reflex discharge with sporadic firing pattern recorded from a singly innervated cell of rat submandibular ganglion in vivo. PMID- 6271411 TI - Modulation of the covalent binding of aryl hydrocarbon metabolites to DNA in vitro after treatment of rats and mice with trans-stilbene oxide. AB - The effect of trans-stilbene oxide (TSO) induction on the microsome-catalyzed binding of polycyclic aromatic hydrocarbon metabolites to DNA was investigated using two rodent species (Sprague-Dawley rat and C57BL/6N or NMRI Swiss mouse) and 2 different binding substrates (benzo[a]anthracene). It was determined that TSO exerts 2 separate effects on polycyclic aromatic hydrocarbons - it increases the rate of oxidation at the K-region of the molecule due to its induction of specific monooxygenases, and it increases the rate of deactivation of epoxide intermediates by induction of microsomal epoxide hydrolase activity. The importance of these individual effects were determined by inducing monooxygenase activity with BP, altering region specificity and inducing epoxide hydrolase (EH) activity with TSO, assessing the combined inductive effects of TSO and BP, inhibiting EH with 1,1,1-trichloropropene oxide, or increasing its activity by the addition of pure enzyme. This study shows that these effects are similar for both substrates examined, and that the effect of TSO on the binding to DNA of highly carcinogenic bay-region diol-epoxides is multi-faceted, due to its multiple inductive effects. PMID- 6271412 TI - Metabolism of benzo[a]pyrene in rat prostate glands following 2,3,7,8 tetrachlorodibenzo-p-dioxin exposure. PMID- 6271413 TI - Lung and liver cell-mediated mutagenesis systems: specificities in the activation of chemical carcinogens. AB - A liver and lung cell-mediated-V79 cell mutagenesis system using intact cells as metabolic activation systems was employed to study the relative ability of cells from these organs to activate chemical carcinogens. Primary cultures of liver and lung cells from male Sprague Dawley rats were used to metabolically activate the chemicals and the mutation of Chinese hamster V79 cells to ouabain resistance used to detect mutagenic intermediates. 7,12-Dimethylbenz[a]anthracene and 3 methylcholanthrene, were more active in the lung system than in the liver cell system. Benzo[a]pyrene (B[a]P) was inactive in the liver cell-mediated system but mutagenic to V79 cells in the lung cell-mediated system. Dimethylnitrosamine (DMN) was inactive in the presence of liver cells. Aflatoxin B1 was mutagenic in the liver cell-mediated system, but only weakly mutagenic in the lung cell mediated system. Because the mutagenicities of DMN and B[a]P were organ-specific, the metabolism of these carcinogens in the two primary cell systems was investigated. DMN was metabolized by liver but not by lung cells, possibly accounting for its lack of mutagenicity in the lung cell system. B[a]P was extensively metabolized by both cell types, but quantitative differences were observed when the metabolic products were analyzed by high pressure liquid chromatography. Comparing total organic and water soluble metabolites, lung cells produced similar amounts of 7,8- and 9,10-diols but little 4,5-diol, while liver cells produced equivalent total amounts of the three diols. Lung cells produced twice the amount of B[a]P glucuronide conjugates as liver cells, while liver cells produced twice the amount of B[a]P sulfate conjugates as lung. The data suggest that intact cells from various organs can be used as metabolic activating systems in vitro assays and that studies into organ specificity can be investigated by this approach. PMID- 6271414 TI - DNA methylation and methylase levels in normal and malignant mouse hepatic tissues. AB - The status of DNA methylation, as measured by the 5-methylcytosine content of nuclear DNA, was examined in normal livers and in chemically induced or spontaneous primary hepatocellular carcinoma (PHC) arising in three strains of mice. The DNA from spontaneous tumors of genetic origin in C3H mice and also from acetylaminofluorene, chlordane, or 3'-methyl-4-dimethylaminoazobenzene-induced tumors in C57Bl and B6C3 mice was undermethylated compared to the levels in background and normal liver samples. The DNA methylase activities from normal liver, background liver, and PHC were assayed in C3H mice to determine whether the observed genomic undermethylation is related to a dysfunction of this enzyme and were compared to the rates of DNA synthesis in these tissues. Since DNA methylase levels from tumor nuclei were elevated compared to background, it is concluded that the undermethylation found in the tumor genomes of this system is not due to inactivation nor a significant deficiency of the activity of this enzyme relative to the demand in tumors for methylation of de novo synthesized DNA. PMID- 6271416 TI - Metabolism of benzo[a]pyrene by brain microsomes of fetal and adult rats and mice. Induction by 5,6 benzoflavone, comparison with liver and lung microsomal activities. AB - Using brain, lung and liver microsomes as the enzyme source in in vitro assays, benzo[a]pyrene (B[a]P) metabolism was studied in fetuses and dams of mice (C57B1/6) and rats (WAG). Separation and quantitation of B[a]P metabolites were performed by h.p.l.c. Microsomal preparations were tested for cytochrome P-450 dependent O-dealkylation of 7-ethoxycoumarin and epoxide hydrolase activities. Another parameter measured included the conjugation of 1-chloro-2,4 dinitrobenzene to glutathione by cytosolic glutathione-S-transferase activity. The induction of B[a]P metabolism was studied after treatment of animals with 5,6 benzoflavone (BF). Mixed function oxygenase, epoxide hydrolase and glutathione-S transferase activities were transplacentally inducible after dams were treated with BF. Metabolic activation of B[a]P by fetal brain microsomes was lower in both species than that by fetal lung and liver microsomes, but it was higher in fetuses than in adults. All metabolites of B[a]P increased after BF treatment; the production of 7,8-dihydro-7,8-dihydroxybenzo[a]pyrene (7,8-dihydrodiol B[a]P) was higher in brain microsomes from BF-treated rats than that in mice. In stimulated rats, the formation of 7,8-dihydrodiol B[a]P by fetal brain microsomes were higher than that by fetal lung microsomes, whereas in mice, the opposite was observed. These data suggest that initiation could occur in utero, and partially explain the species-specific differences in susceptibility to transplacental tumorigenesis by polycyclic aromatic hydrocarbons by differences in biotransformation in the target organ. PMID- 6271417 TI - Loss of the glomerular contractile response to angiotensin in rats following myohemoglobinuric acute renal failure. AB - To examine the state of the glomerulus in rats protected from acute renal failure (ARF) by prior insult, we measured the contractile responses of isolated glomeruli to angiotensin II (AII) and dibutyryl cyclic AMP (DBcAMP). In healthy rats, both agents induced a dose-related fall in glomerular diameter (P less than 0.001). Saralasin, the angiotensin antagonist, blocked the glomerular response to AII totally and to DBcAMP partially. Two weeks following ARF induced with 50% glycerol (10 ml/kg, im), azotemia had reversed and the nephrotoxic effect of mercuric chloride (4.7 mg/kg, sc) was blunted, as anticipated. AII did not reduce glomerular size, but the response to DBcAMP was sustained at this time. To determine the specificity of the loss of the glomerular response to AII, we also assessed the effects of an AII infusion (1 microgram/kg/min) on blood pressure and renal blood flow. There was a substantial rise in blood pressure (control, 104 +/- 11.8 mm Hg; AI, 131.0 +/- 5.3 mm HG; P less than 0.001) and fall in renal blood flow (control, 2.45 ml/g per min; AII, 0.81 +/- 0.10 ml/g per min; P less than 0.025). Vascular responsiveness to AII was preserved at a time when glomeruli were totally unresponsive and rats were resistant to ARF. The loss of glomerular contractility may, in part, account for the protection from ARF seen in this model: if so, glomerular abnormalities may play a pathogenetic role. PMID- 6271415 TI - Correlation between preneoplastic lesions in rat embryo lung treated with B[a]P or CSC in organ culture and tumour production in vivo. AB - The purpose of this study was to follow the evolution of normal and tested rat embryo lungs in organ culture and to classify the induced lesions in relation to the in vivo tumour histogenesis. Lungs from 15 day old Wistar rat embryos were maintained in organ culture in M 199 medium and 20% horse serum and treated with benzo[a]pyrene (B[a]P) or cigarette smoke condensate (CSC). Explants were studied 30 days of culture. The control explants display a prevalence of epithelial tissue along with a reduction in connective tissue, whereas treated explants show similar morphological alterations after both B[a]P and CSC. The percentage of the observed lesions depends on the strain of rats used. Alterations are classified into preneoplastic stages according to Shabad's pattern developed for rat lung tumours in vivo. This classification permits the assessment and quantitation of the carcinogenic effect of a substance. To prove that lung explants may undergo neoplastic transformation after treatment in vitro, normal and carcinogen-treated lung organ explants were dispersed and, after cell subculture, injected i.p. into isogenic rats. Tumours were obtained after only 5 subcultures with treated cells, whilst with the controls, tumours were obtained after 21 subcultures. PMID- 6271418 TI - Angiotensin I recovery from plasma incubated with organic reagents. AB - (1) In order to explore the possible mechanism that organic reagents used in the incubation step of the plasma renin activity (PRA) analysis act as a angiotensinase inhibitors we did angiotensin I (AI) recovery studies from plasma with such reagents. The organic acids and their mean difference in percent recovery of AI as compared to that for hydrochloric acid (HCI) are respectively as follows for one and a three hour incubation time: maleic (0.7%, 4.5%); and potassium hydrogen phthalate (KHphthalate) (7.2%, 9.6%). The tris(hydroxymethyl)aminomethane (Tris) organic acid salts and their mean difference in percent recovery of AI as compared to that for Tris-HCl are as follows for a one hour incubation: Tris-acetylsalicylate (3.6%), Tris phenoxyacetate (3.6%), Tris-benzoate (2.6%), and Tris-salicylate (4.9%). (2) Of the reagents studied KHphthalate after a three hour incubation produced a statistically significant difference from the HCl reagent. The recovery data for all the organic reagents suggested that the primary mechanism of action was not that of an angiotensinase inhibitory one. PMID- 6271419 TI - Heterogeneity of corticotropin-immunoreactive compounds in human body fluids. AB - Using reversed-phase "high-performance" liquid chromatography and hydrophilic ion pairing reagents, we studied the chromatographic profiles of corticotropin immunoreactivities in human plasma, cerebrospinal fluid, and urine. The chromatographic system we applied provides excellent separation of peptide molecules, with reproducibility (CV) of retention times better than 0.5%, and makes feasible the subsequent immunological quantitation of eluted fractions. Profiles of corticotropin immunoreactivities exhibited several chemically different compounds in each of the three body fluids studied. A considerable portion of these immunoreactive compounds is more hydrophilic and probably of lower molecular mass than the human corticotropin 1-39 molecule itself. These hydrophilic molecules are more predominant in urine and plasma than in cerebrospinal fluid. PMID- 6271420 TI - Colorimetry of angiotensin-I converting enzyme activity in serum. AB - This simple, accurate, and reproducible colorimetric method for determining the activity of angiotensin-I converting enzyme is based upon colorimetry of the quinoneimine dye produced from the substrate p-hydroxyhippuryl-L-histidyl-L leucine by action of this enzyme through the following series of reactions. The enzyme acts on the substrate to yield p-hydroxyhippuric acid and L-histidyl-L leucine. The former is then hydrolyzed in the presence of hippuricase to produce p-hydroxybenzoic acid and glycine. Finally, oxidative coupling of p hydroxybenzoic acid with 4-aminoantipyrine is catalyzed by peroxidase in the presence of hydrogen peroxide, producing a quinoneimine dye, the concentration of which is measured at its absorbance maximum at 505 nm to evaluate the activity of ACE. The Km value for the above-mentioned substrate is 0.32 mmol/L, the optimum pH is 8.3. Results by the present method and Cushman and Cheung's method (Biochem. Pharmacol. 20: 1637, 1971) correlate closely (r = 0.986). The within run CV is 2.93%. PMID- 6271421 TI - Adaptation of a NADH-linked serum 5'-nucleotidase procedure to a small benchtop centrifugal analyzer. PMID- 6271422 TI - An extraordinarily long survival of a patient with inoperable cancer of the breast. PMID- 6271423 TI - Serum apoprotein A-I and A-II levels in liver diseases and cholestasis. AB - Serum apoprotein A-I and A-II levels were determined by electroimmunoassay in patients with liver diseases and cholestasis. Significant decreases in apoprotein A-I and A-II levels were observed in such patients. The decreases were especially pronounced in the early phase of acute hepatitis and cholestasis. The decreases in A-II levels were more prominent than the decreases in A-I in severe hepatic dysfunction or cholestasis. Accordingly, the A-I/A-II ratio showed no change in the convalescent phase of acute hepatitis or chronic hepatitis but increased significantly in the early phase of acute hepatitis, cirrhosis of the liver, hepatoma, and cholestasis. The results suggested the existence of a high density lipoprotein with an abnormal apoprotein composition or a more profound decrease of HDL3 than of HDL2 in severe hepatocellular dysfunction of cholestasis. PMID- 6271424 TI - Urinary excretion patterns of cyclic AMP and cyclic GMP following thermal injury. AB - Measurement of 24-h urinary cyclic AMP and cyclic GMP levels in 19 patients for up to nine days after thermal injury has revealed differences in cyclic nucleotide excretion patterns between "severe" and "mild to moderate" groups of burned patients, classified according to a predictive index of burn mortality. Cyclic AMP excretion fell significantly from a high initial level in the "severe" group, but showed no significant change in the "mild to moderate" group. Differences in cyclic AMP excretion between the two groups of patients were only significant on the second day following burn injury. There was no correlation between cyclic AMP output averaged for each patient over the first nine days and percentage body surface area of the burn or the predictive index. In contrast to the results for cyclic AMP excretion, cyclic GMP output rose significantly over the 9-day period in severely burned patients, while levels reached a plateau on days four to six for the moderate to mild group. When cyclic GMP excretion was averaged over the first nine days for each patient, a significant correlation with both percent body surface area of burn and the predictive index of burn mortality was found. The tissue source or sources which are responsible for increased cyclic GMP excretion are as yet unknown. PMID- 6271425 TI - Do conventional plasma cyclic nucleotide phosphodiesterase inhibitors really work in all situations? PMID- 6271426 TI - Altered endocrine response to partial thyroidectomy in propranolol-prepared hyperthyroid patients. AB - The endocrine response to partial thyroidectomy in a group of twenty hyperthyroid patients prepared with propranolol alone was compared to that of a matched control group of ten euthyroid patients. In propranolol-prepared patients the glucose response to surgery was reduced (P less than 0.05) for up to 4 h post operatively and biochemical hypoglycaemia was noted in one patient. Both thyroxine and triiodothyronine (T3) fell significantly, associated with a marked rise in reverse T3. Growth hormone levels were higher (P less than 0.05) both pre and post-operatively in propranolol-prepared patients, whereas prolactin levels, although similar pre-operatively, were lower (P less than 0.05) in these patients post-operatively. Cortisol and ACTH levels were lower (P less than 0.05) both before and following thyroidectomy in propranolol-prepared patients. These results suggest that the endocrine response to surgical stress is markedly altered in propranolol-prepared hyperthyroid patients. PMID- 6271427 TI - [The islet cell tumors of the pancreas (author's transl)]. PMID- 6271428 TI - Different populations of rheumatoid factor idiotypes induced by two polyclonal B cell activators, pokeweed mitogen and Epstein--Barr virus. PMID- 6271429 TI - Cell-mediated immunity to homologous basement membrane (type IV) collagen in C57BL/6 mice. PMID- 6271430 TI - Experiences with recurring ventricular catheter obstructions. AB - Seven cases are presented of recurrent ventricular catheter obstruction, an infrequent but serious problem in the management of hydrocephalus. Plugging of the catheter was caused by detritus rather than by choroid plexus. A retrospective analysis of 214 shunt revisions indicated that obstruction both by detritus and by choroid plexus were incidental phenomena in the great majority of the cases - the former type occurring mainly within the first month, the latter between 3 and 6 months after operation. It is suggested that the ultimate cause of recurrent catheter obstruction by detritus may be destruction or ablation of the ependymal lining of the ventricles (e.g. by an inflammatory process), which easily provides debris to the catheter. In 4 of the cases insertion of the catheter into the opposite ventricle prevented further recurrence, when reinsertion into the same ventricle had been ineffective. In one case, with scarcely dilated ventricles, external decompression was helpful. In patients with compartmentalisation of the ventricles (substantiated in one of our patients) one might consider removal of the septa via craniotomy before the insertion of the catheter. PMID- 6271431 TI - Acute psycho-organic reactions after cervical myelography using metrizamide. PMID- 6271432 TI - Organic psychosyndrome correlated with high density of grey matter on CT following metrizamide cervical myelography. PMID- 6271433 TI - Congenital fibre type disproportion. AB - Four children with congenital fibre type disproportion were described. It was shown that their type 1 fibres were at least 12% smaller than the type 11 fibres. There was no increase in the terminal innervation ratio (TIR), but a decreased number of terminal knobs was observed in the biopsy of one child. The distribution of fibre types in the biopsy of another child bears out the notion that the abnormalities as seen in the biopsy can be traced back to the spine. PMID- 6271434 TI - Cranial subdural metastases: a clinicopathological study. AB - During a period of 10 years, 2782 brains were examined at post-mortem and in 17 cases metastases were attached to the dura mater, 8 presenting as multiple nodules 6 as a single nodule and 3 as diffuse dural thickening. The primary growths were, in equal proportions sarcomas, epitheliomas and lymphoid tumours. In contrast to the frequent location of metastases in the axial skeleton (9 cases), brain involvement was rarely observed (2 cases). The clinical findings were non-specific. The difference in biological behaviour between subdural and intracerebral metastases is stressed. PMID- 6271435 TI - Migraine with interparoxysmal sharp complexes in the EEG: clinical features and response to anticonvulsant drugs. AB - The clinical profile and therapeutic response of 25 patients with migrainous headaches with interparoxysmal electroencephalographic (EEG) abnormalities in the form of sharp complexes are discussed. The clinical features of these patients consisted of male preponderance, early age of onset in males, long history of illness in females, very low incidence of aura and family history of migraine, frequent severe paroxysms, poor response to routine anti-migraine drugs and good response to anti-convulsant drugs. It is speculated that the EEG abnormalities in migraine may have a congenital basis which finds expression in vasomotor instability through hypothalamus and autonomic nervous system. PMID- 6271436 TI - Arteriovenous malformations of the choroid plexus operative technique. PMID- 6271437 TI - Visualization of cutaneous hemangioma with Tc-99m tagged red blood cells. AB - Scintigraphy with Tc-99m labeled red blood cells (RBCs) was used to evaluate a patient with a large cutaneous hemangioma. The usefulness of this procedure when combined with arteriography is discussed. PMID- 6271438 TI - Ulcerative colitis and a bleeding polyp detected on Tc-99m-pertechnetate abdominal scintigraphy. AB - Two children with rectal bleeding were diagnosed as having ulcerative colitis and a bleeding colonic polyp respectively using abdominal scanning with Tc-99m pertechnetate. Early flow studies are recommended with careful attention paid to the amount of time abnormal areas of activity are seen on the abdominal scan. PMID- 6271439 TI - Assessment of cardiac wall motion with the ejection fraction image: a comparison with contrast left ventriculography. AB - Twenty patients with ischemic heart disease were studied with biplane contrast left ventriculography and gated bloob pool scans. An ejection fraction (EF) image was calculated from each gated blood pool scan. The EF image and contrast ventriculograms were divided into three regions and seven segments respectively. The sites of asynergy observed in each study were compared. Segments two, three and six of the contrast ventriculogram corresponded to the anteroseptal and inferoapical regions of the EF image, but it was difficult to differentiate between these segments on the EF image. Segments three and four corresponded to the inferoapical region and segments five and seven corresponded to the posterolateral region. Diffuse asynergy with a low EF (less than 30%) causes a large defect on the EF image. The mean regional EF obtained from the EF image correlated well with the EF calculated from the left ventricular volume curve (n = 50, r = 0.94). PMID- 6271440 TI - "Mottled" liver scan in giant hepatomegaly due to intrasinusoidal metastasis of small cell lung cancer. PMID- 6271441 TI - Single-pass ventricular function studies. PMID- 6271442 TI - Renal function and imaging studies. PMID- 6271443 TI - Gallium scanning in inflammatory and neoplastic conditions. PMID- 6271444 TI - Optimizing the performance and interpretation of bone scans. PMID- 6271445 TI - Radionuclide imaging in the era of computed tomography. PMID- 6271446 TI - False positive dynamic imaging of the cerebral circulation due to a congenital anomaly. AB - False positive dynamic imaging of the cerebral circulation simulating vascular occlusive disease resulted from a clinically insignificant congenital variation in several patients at our institutions. The anomaly consisted of the asymmetrical caliber of the internal carotid and proximal anterior cerebral arteries. The 1 mm difference in the diameter of the internal carotid arteries was sufficient to be appreciated on dynamic imaging. The frequency of this anomaly was found to be 5% in a series of 200 bilateral carotid angiograms. PMID- 6271447 TI - Prescribing in pregnancy. Corticosteroids. PMID- 6271448 TI - Radiographic findings and etiologic diagnosis in ambulatory childhood pneumonias. AB - The chest roentgenograms of 128 consecutive ambulatory children with radiologic pneumonia were read independently and without clinical information by a faculty general pediatrician (Ped), a pediatric radiologist (R-P) and a general radiologist (R-G). The films were classified as normal, indicative of a viral or bacterial process, or indeterminate. Readings were compared with results of viral titers and bacterial cultures. Agreement between any two observers in classifying films, measured by unweighted Kappa, while statistically significant (p less than 0.001) for any pair, was low. There was no significant difference between the agreement scores of Ped/R-P, Ped/R-G, and R-P/R-G. Twenty-one patients had fourfold viral titer increases (N = 16) or positive bacterial cultures of blood or pulmonary aspirate (N = 5). The sensitivity of viral readings for titers increases varied from 19% to 68% depending on observer type; the sensitivity of bacterial readings for positive bacterial cultures varied from 60% to 80%. The three observers agreed on a correct reading in only three children with viral and three with bacterial pneumonia. Because of poor observer agreement and appreciable false-negative errors when viral and bacterial readings were compared to titer increases and positive bacterial cultures, respectively, we conclude that radiographic findings are poor indicators of etiology diagnosis in ambulatory childhood pneumonias and, of themselves, are an insufficient data base for making therapeutic decisions. PMID- 6271450 TI - Characterization of collagenase, other metallo-proteinases and an inhibitor (TIMP) produced by human synovium and cartilage in culture. AB - 1. Explants of human synovium from normal, rheumatoid and osteoarthritic subjects produced proteinases in culture that could degrade connective tissue macromolecules at neutral pH. 2. The proteinases detected in the media were predominantly of the class requiring metal ions for activity, and occurred in either partially or wholly latent forms. Activation could be achieved by treatment with either trypsin or the organo-mercurial, 4-aminophenylmercuric acetate. 3. Explants of both normal and osteoarthritic synovium produced an inhibitor of collagenase in the early days of culture, similar to that previously described for rabbit tissues. In contrast, no free inhibitor could be detected in the media of rheumatoid synovial cultures. 4. Explants of normal human articular cartilage produced latent collagenase and free inhibitor in culture. 5. These findings suggest that the activity of tissue-derived metallo-proteinases may be controlled by locally synthesized inhibitors, by mechanisms analogous to those already described for rabbit model systems. Changes in the synthesis and degradation of inhibitor could be important in human diseases in which destruction of connective tissue is a prominent feature. PMID- 6271449 TI - Production of collagenase and inhibitor (TIMP) by normal, rheumatoid and osteoarthritic synovium in vitro: effects of hydrocortisone and indomethacin. AB - 1. The amounts of latent and active collagenase and of collagenase inhibitor (TIMP) produced by two normal, three rheumatoid and two osteoarthritic synovial specimens in culture were compared. Normal synovia produced TIMP, but little latent enzyme. Rheumatoid synovia produced higher levels of total collagenase activity than normal, of which up to 50% in one sample was present in the medium in an active form, whereas no specific inhibitory activity due to TIMP was detectable. The amounts of collagenase and TIMP produced by osteoarthritic synovia were more variable and appeared to reflect the degree of inflammation in the tissue at the time of initiating the cultures. 2. Concentrations of TIMP were usually higher in the culture media of normal, rheumatoid and osteoarthritic synovia when hydrocortisone was present. Correspondingly, amounts of total collagenase were reduced. Production of prostaglandin E (PGE) were inhibited in a dose-dependent manner by hydrocortisone. 3. Indomethacin had no consistent effect on the production of TIMP by rheumatoid and osteoarthritic synovia, although it tended to depress production of collagenase. The production of TIMP by normal synovia was depressed by indomethacin. No PGE was detectable in the media when indomethacin was present. 4. These results are consistent with those from previous animal studies, and we conclude that the balance between production of collagenase and TIMP may be critical in determining the extent of the destructive processes in arthritis. The ability of hydrocortisone to suppress production of collagenase and to increase free TIMP concentration, as well as to inhibit synthesis of prostaglandin, may explain in part how the drug exerts its therapeutic effects in patients with rheumatoid arthritis. PMID- 6271451 TI - Some general questions about movable elements and their implications. PMID- 6271452 TI - Structural analysis of Tn5. AB - Nucleotide sequences have been determined for the 1.5-kb inverted repeats of Tn5 and for their junctions with the central unique region and with host DNA. The primary findings stemming from this analysis are: 1. Integration of Tn5 is accompanied by the duplication of 9 bp of host DNA. 2. Loss of Tn5 occurs by crossover between short, homologous nucleotide sequences near the junction between Tn5 and host DNA. 3. The IR sequences contain long, open translational reading frames that may code for transposase proteins. 4. The two IR sequences differ by a single-base change. This alteration accounts for the two functional differences observed between IRL and IRR: It shortens the reading frame for the transposase gene in IRL, and it improves the efficiency of a promoter for the nearby Km-resistance gene. 5. The NH2 terminus of the structural gene for the Km resistance gene maps at the very left border of the unique region, i.e., very close to the end of IRL. These results support the view that the inverted repeats of Tn5 stem from two identical copies of an originally independently moving DNA element. In the transposon, one of these, IRL, seems to have evolved toward a close physical and functional linkage with the antibiotic-resistance gene. PMID- 6271453 TI - Trans-acting genes of bacteriophages P1 and Mu mediate inversion of a specific DNA segment involved in flagellar phase variation of Salmonella. PMID- 6271454 TI - Insertion, excision, and inversion of Tn5. PMID- 6271455 TI - Analysis of the structure and function of the kanamycin-resistance transposon Tn903. PMID- 6271456 TI - Identification of a sex-factor-affinity site in E. coli as gamma delta. PMID- 6271457 TI - Internal rearrangements of IS2 in Escherichia coli. PMID- 6271458 TI - Activation of gene expression by IS2 and IS3. PMID- 6271459 TI - IS1-promoted events associated with drug-resistance plasmids. PMID- 6271460 TI - Intramolecular transposition of beta-lactamase sequence and related genetic rearrangements. PMID- 6271461 TI - Analysis of the functional components of the phase variation system. PMID- 6271462 TI - Detection of replicational inceptor signals in IS5. PMID- 6271463 TI - Genes are things you have whether you want them or not. PMID- 6271464 TI - Transposition immunity. PMID- 6271465 TI - Regulation of Tn5 transposition. PMID- 6271466 TI - Mutants of Escherichia coli affected in the processes of transposition and genomic rearrangements. PMID- 6271467 TI - Isolation of polA mutation that affects transposition of insertion sequences and transposons. PMID- 6271468 TI - Substitution of silent bacterial genes by a bacteriophage lambda variant carrying IS1. PMID- 6271469 TI - Studies on transposition mechanisms and specificity of IS4. PMID- 6271470 TI - Genetic organization of Tn10 and analysis of Tn10-associated excision events. PMID- 6271471 TI - Recombination involving transposable elements: on replicon fusion. PMID- 6271472 TI - Genetic and sequencing studies of the specificity of transposition into the lac region of E. coli. PMID- 6271473 TI - Tn3 encodes a site-specific recombination system: identification of essential sequences, genes, and the actual site of recombination. PMID- 6271474 TI - Tn3: transposition and control. PMID- 6271475 TI - Genesis and natural history of IS-mediated transposons. AB - The natural genesis of IS1-mediated transposons containing the genetic determinant cat for chloramphenicol resistance is documented. First, the small plasmid pBR325 containing the cat gene served as a target in IS1-mediated transpositional cointegration with the genome of bacteriophage P1, which was the source of the IS1. From the resulting pBR325:P1 plasmids, pBR325::IS1 segregants were isolated. Upon growth of a phage lambda derivative in the presence of this plasmid, rare plaque-forming lambda Cmr specialized transducing phages were formed. In each of six independent lambda Cmr isolates studied, the cat gene was carried between flanking IS1 elements. In one case, these IS1 elements were in the same orientation; in the other five cases, they were in opposite orientation. All of these IS1-cat-IS1 structures transposed as units to the genome of phage P1 15, pointing to stable maintenance of the transposon. However, appropriate selection allowed us to follow the decay of these transposons. Models to explain the genesis of transposons with directly and inversely repeated IS elements are discussed, as well as the evolutionary implications of these mechanisms. PMID- 6271476 TI - Transposon-specified, site-specific recombination systems. PMID- 6271477 TI - Mechanism of insertion and cointegration mediated by IS1 and Tn3. PMID- 6271478 TI - Mechanism of bacteriophage Mu DNA transposition. PMID- 6271479 TI - Transposition studies using a ColE1 derivative carrying bacteriophage Mu. PMID- 6271480 TI - Two pathways in bacteriophage Mu transposition? PMID- 6271481 TI - Transposition of bacteriophage Mu: properties of lambda phages containing both ends of Mu. PMID- 6271482 TI - Genetic study of Mu transposition and Mu-mediated chromosomal rearrangements. PMID- 6271483 TI - Specificity of bacteriophage Mu integration into DNAs of different origins. PMID- 6271484 TI - DNA gyrase: site-specific interactions and transient double-strand breakage of DNA. PMID- 6271486 TI - Instability of palindromic DNA in Escherichia coli. PMID- 6271485 TI - In vitro study of illegitimate recombination: involvement of DNA gyrase. PMID- 6271487 TI - Strand exchange in lambda integrative recombination: genetics, biochemistry, and models. AB - We have asked, "What is the mechanism of strand exchange during site-specific recombination of phage lambda?" Crosses carried out in vivo have shown that the recombination joint can be extended rather than flush and that the four-strand breaks and rejoinings needed to from a recombinant can occur asynchronously. Crosses carried out in vitro have shown that all the nucleotides at the site of crossover are conserved during recombination, as are most or all of the superhelical turns present in the substrate molecules. We have presented new data showing that topoisomerase activity of Int protein relaxes DNA by making transient single-strand, rather than double-strand, breaks in the phosphodiester back-bone. These findings are incorporated into a model for strand exchange that has as its central intermediate a four-strand structure. PMID- 6271488 TI - Regulation of the integration-excision reaction by bacteriophage lambda. PMID- 6271489 TI - Transposons encoding trimethoprim or gentamicin resistance in medically important bacteria. PMID- 6271490 TI - A study of the dissemination of Tn1681: a bacterial transposon encoding a heat stable toxin among enterotoxigenic Escherichia coli isolates. PMID- 6271491 TI - Basis of transposition and gene amplification by Tn1721 and related tetracycline resistance transposons. PMID- 6271492 TI - Hitchhiking transposons and other mobile genetic elements and site-specific recombination systems in Staphylococcus aureus. PMID- 6271493 TI - Evidence for conjugal transfer of a Streptococcus faecalis transposon (Tn916) from a chromosomal site in the absence of plasmid DNA. PMID- 6271494 TI - Inverted-repeat nucleotide sequences in Escherichia coli and Caulobacter crescentus. PMID- 6271495 TI - ISR1: an insertion element isolated from the soil bacterium Rhizobium lupini. AB - The insertion element ISR1 was isolated from the soil bacterium R. lupini. In this strain, ISR1 shows a very strong affinity to plasmid RP4. It causes RP4 mutations at the strikingly high frequency of 10(-2) to 10(-1), either by the integration itself or by generating deletions. In E. coli, ISR1 seems to be inactive. No evidence could be obtained for a promoter site on ISR1 or for an ISR1-encoded protein. Our results indicate, however, an ISR1-specific termination signal for either transcription or translation. PMID- 6271496 TI - IS1-mediated DNA rearrangements. PMID- 6271497 TI - Genetic organization of Tn5. PMID- 6271498 TI - Alpha-adrenergic receptor function in schizophrenia, affective disorders and some neurological diseases. PMID- 6271499 TI - Cyclic nucleotides in anatomical subdivisions of the bovine lens. AB - Adenosine 3', 5'-cyclic monophosphate (cAMP) and guanosine 3', 5'-cyclic monophosphate (cGMP) have been measured in whole and anatomically subdivided areas of bovine lenses. The highest values of cAMP and cGMP (2.843 pmoles/mg protein and 0.0139 pmoles/mg protein respectively) were found in the epithelium. In contrast, the lowest values of cAMP and cGMP (.007 pmoles/mg protein and .001 pmoles/mg protein respectively) were detected in the central nucleus. Values for the cyclic nucleotides in the whole tissue more closely resembled those of the whole nucleus. PMID- 6271500 TI - Complement-dependent cytotoxicity in rats bearing human adenovirus type 12 induced primary retinoblastoma-like tumor in the eye. AB - Using an animal model of retinoblastoma in inbred rats and cultured human adenovirus type 12-induced retinoblastoma-like tumor cells (RAO 188), complement dependent cytotoxicity was determined by measuring release of 3H-uridine labelled RNA. Sera from rats in which tumors did not grow after adenovirus type 12 inoculation had higher cytotoxicity against RAO 188 cells than sera from rats bearing primary adenovirus type 12-induced retinoblastoma-like tumor. These results showed that the rat which could raise antibodies against adenovirus type 12-induced retinoblastoma-like tumor cells did not allow the tumor growth in the eye after virus inoculation. PMID- 6271501 TI - Ultrastructural cytochemistry of the ocular lens. AB - Ultrastructural and cytochemical investigations of ocular lens have been difficult due to limited applicability of currently available techniques for the preparation of this tissue for such studies Several modifications in currently available techniques are being made in order to preserve the fine structure of the lens. In this report we present a modified method which provides good preservation of lens fine structure and allows cytochemical localization of enzymes in this tissue. Brief freezing permits preparation of thin slices of the lens which are subsequently incubated in the appropriate media for enzyme cytochemistry at an ultrastructural level. We have used this modified method for the localization of Na-K-dependent ATPase and the results are presented in this report. PMID- 6271502 TI - Herpes simplex virus type 2 and cervical cancer. PMID- 6271503 TI - Perinatal listeriosis. Tolerance of a clinical isolate of Listeria monocytogenes for ampicillin and resistance against cefotaxime. AB - A strain of Listeria monocytogenes isolated from a case of early onset, generalized, fatal, perinatal listeriosis in a premature infant was found to be tolerant for ampicillin and resistant against cefotaxime. A laboratory control strain of L. monocytogenes and Staphylococcus aureus control strain ATCC 25923 proved tolerant for both beta-lactam antibiotics, respectively. The combinations of ampicillin with gentamicin and cefotaxime with gentamicin resulted in additive bactericidal activity against both strains of L. monocytogenes. Based on these findings and those of the literature, it is suggested that cases of systemic listeriosis be treated with a combination of a beta-lactam antibiotic, preferably ampicillin, with an appropriate aminoglycoside, such as gentamicin. PMID- 6271504 TI - [Clinical analysis of 24 cases of alveolar cell carcinoma of bronchiolar origin (author's transl)]. PMID- 6271505 TI - Indirect and direct stimulation of fatigued human muscle. PMID- 6271506 TI - Glycolytic and oxidative energy metabolism and contraction characteristics of intact human muscle. AB - It is proposed that glycolytic rate may be measured as lactate accumulation after electrical stimulation of the quadriceps femoris muscle under anaerobic conditions. The ratio of glucose 6-phosphate to lactate is an internal monitor of the glycolytic pathway. The phosphocreatine/lactate ratio links glycolysis and the creatine kinase reaction and could be used to distinguish abnormalities in energy metabolism. The rate of resynthesis of phosphocreatine after stimulation when the circulation is restored should be measure of oxidative phosphorylation. The relaxation rate seems to be a mechanical index of the metabolic state of the muscle. PMID- 6271507 TI - Developmental influences on vascular structure and function. AB - Blood vessels with apparently similar structures show remarkable functional heterogeneity. Differences exist in the nature and extent of their innervation, synaptic architecture, receptor characteristics, excitation coupling systems, capacity for intrinsic tone, contractility, elasticity and calcium-pool dependence, to mention only a few variables. The underlying basis of these differences is unknown. Similarity between the distribution of particular features in the vascular bed and the early patterns of embryological development suggest that some specific functional characteristics are determined during the process of gastrulation, if not before. Examples are given of receptor characteristics and tissue sensitivity that seem to reflect the different mesenchymal origins of particular vessels. Studies on vessels from immature fetal lambs confirm that individuality of specific vessels is established early. Interruption of sympathetic nerve traffic influences the function of vascular smooth muscle cells in a complex manner which is expressed differently at different ages. If the findings of experiments on the ear artery of the rabbit reveal a general principle seen in the circulation, then the level of sympathetic nerve traffic during growth would be expected to influence both qualitatively and quantitatively the structure and reactivity of the adult circulation. By this effect the sympathetic nervous system exerts a long-term influence on blood pressure. The characteristics of a particular blood vessel in the adult depend on many factors such as the level of blood pressure and the amplitude of the pulse wave which are not discussed in this chapter. These studies show that the character of the mature vessels also reflects influences that occur during early development and growth. PMID- 6271508 TI - Na-K-ATPase in erythrocyte ghosts is not a marker for primary hypertension. AB - In search for a diagnostic marker for essential hypertension, we investigated the activity of the ouabain sensitive Na-K-ATPase and the ouabain insensitive ATPase in erythrocyte ghosts of 57 patients with essential hypertension, 12 patients with renal hypertension, 6 patients with Cushing's syndrome and 4 patients with primary hyperaldosteronism. Na-K-ATPase-activity was increased in patients with essential hypertension and in patients with renal hypertension compared with controls with a considerable overlap. Na-K-ATPase-activity was increased in all patients with Cushing's syndrome but was not different from the control group in patients with Conn's syndrome. Ouabain-insensitive ATPase-activity was similar in all patients and in normotensive controls. The serum of patients with essential hypertension did not exhibit an ouabain-like ATPase-inhibiting activity when incubated with erythrocyte ghosts of normotensive controls. In our hands, determination of the Na-K-ATPase-activity in erythrocyte ghosts cannot be used as a diagnostic marker for essential hypertension. PMID- 6271509 TI - Na countertransport and cotransport in human red cells: function, dysfunction, and genes in essential hypertension. AB - We describe in this paper studies on the modes of operation of ouabain insensitive sodium transport systems in red cells of normotensive and hypertensive patients. We have extensively investigated the properties of Na countertransport and cotransport in order to clarify whether they are two different proteins or one transport protein with two modes of operation. Several criteria of discrimination between the two pathways are described: They differ in their affinity for Na and Li, sensitivity to several inhibitors, changes in cell volume, and chloride replacement by nitrate. We propose that there are two different transport systems. We have found elevated countertransport in red cells of hypertensive patients in France and in the United States. However, the cotransport system was found elevated in patients in Boston but reduced in patients in Paris. Studies of the modes of operation of the Na-K cotransport system indicate that it can promote K accumulation using an inward sodium gradient. This mode might be more efficient than Na extrusion at the physiological level of Na and K gradients. We interpret our findings of elevated Na-K cotransport in American hypertensive patients as an increased number of transport units functioning as K accumulators. It remains to be determined whether the reduced affinity for internal sodium of the outward cotransport is a defective outward cotransport or else a modulation of this transport system to favor K accumulation. PMID- 6271510 TI - Is a circulating sodium transport inhibitor involved in the pathogenesis of essential hypertension? AB - There is controversy about the relationship between sodium intake and the prevalence of high blood pressure. Part of this controversy relates to how an increase in sodium intake could cause an increase in peripheral resistance. We have put forward the following hypothesis. In essential hypertension there is an inherited defect of the kidney's ability to excrete sodium which becomes increasingly obvious the greater the sodium intake. This difficulty in sodium excretion by the kidney increases the concentration of a circulating sodium transport inhibitor that affects sodium transport across many cell membranes. In the kidney the inhibitor adjusts urinary sodium excretion back towards normal so that sodium balance is near that of normal subjects on the same intake of sodium. In the arteriolar smooth muscle the inhibition of sodium transport across the cell wall causes a rise in intracellular sodium concentration which, in turn, raises the intracellular calcium concentration and thus increases vascular reactivity. This hypothesis also proposes that the abnormalities of sodium transport in circulating cells in vivo are directly due to the increased secretion of the circulating sodium transport inhibitor. Evidence supporting this hypothesis is discussed. Firstly, it is pointed out that there is much evidence which suggests that there is a circulating inhibitor of Na+-K+-ATPase, the level of which is related to sodium intake and that the level of this inhibitor appears to be increased in many hypertensives. Secondly, the finding that normotensive white cells incubated in the plasma of hypertensive patients develop the same decrease in the Na+-K+-ATPase dependent sodium transport as the hypertensives own white cells also suggests that hypertensives have an increase in a circulating Na+-K+-ATPase inhibitor. PMID- 6271511 TI - Cation fluxes and (Na+ + K+)-activated ATPase activity in erythrocytes of patients with essential hypertension. AB - Various claims, partially conflicting, have been made in recent years for abnormalities in cation pumps and fluxes in erythrocytes of patients with essential hypertension. In view of the obvious significance of such abnormalities for diagnostic purposes, and possibly for our understanding of the pathophysiology of essential hypertension, we have investigated these claims. We have determined the following parameters of erythrocytes from essential hypertensives and normotensives: 1. (Na+ + K+)-ATPase activity and the Km values for Na+, K+ and ATP; 2. ouabain-sensitive fluxes of Na+ and K+ in Na+-enriched cells after cold treatment and after treatment with p chloromercuribenzenesulphonate; 3. furosemide-sensitive, ouabain-insensitive cotransport efflux of Na+ + K+. No significant differences were observed, except for a slight decrease in the ouabain-sensitive K+ influx after cold treatment in hypertensives. Hence, we conclude that determination of these parameters in erythrocytes does not seem to be useful for the diagnosis of essential hypertension. PMID- 6271512 TI - A genetic approach to the geography of hypertension : examination of Na+ - K+ cotransport in Ivory Coast Africans. AB - Outward Na+ - K+ cotransport in erythrocytes from essential hypertensive Caucasian subjects was found to be excessively low (Co -) compared to normotensives (Co +) carefully selected for their negative family history of hypertension. Since the frequency of essential hypertension varies widely among different populations and is particularly high in certain coloured peoples, we compared erythrocyte Na+ - K+ cotransport in normotensive and hypertensive subjects in Paris (France) and in Abidjan (Ivory Coast) to seen whether defective cotransport was related to high blood pressure in the African group as well. Of the 66 French unselected normotensives investigated, 26 (39%) were Co - whereas 14 of the 18 Ivory Coast unselected normotensives (79%) were Co -. 64 (80%) of the 80 essential hypertensives examined in France were Co -, but the proportion of Co - subjects among the Ivory Coast hypertensives was even higher. In addition, both hypertensives and normotensives in the African groups often had undetectable outward Na+ effluxes, a rare finding in the French subjects. We suggest that the high incidence of abnormal Na+ - K+ cotransport in the Ivory Coast series may reflect a genetic propensity to hypertension in this population, and that consequently, Na+ - K+ erythrocyte cotransport measurements might prove useful in defining geographic variations in hypertension. PMID- 6271513 TI - Furosemide and bumetanide-sensitive Na+ fluxes in erythrocytes from genetically hypertensive rats (SHR). AB - Na+ and K+ fluxes from erythrocytes of SHR and WKY control rats have been studied. Fluxes were measured in both fresh and sodium-loaded (after exposure to ouabain or PCMBS) erythrocytes. Pump activity appears higher and furosemide or bumetanide sensitive Na+ efflux lower in SHR compared to WKY suggesting a similar abnormality in SHR genetic hypertension as in human hypertension. PMID- 6271514 TI - Enzyme kinetic studies in cell populations using fluorogenic substrates and flow cytometric techniques. PMID- 6271515 TI - Perianal Paget's disease associated with cloacogenic carcinoma: report of a case. AB - A case of perianal Paget's disease associated with a cloacogenic carcinoma is presented. There was no clinical evidence of the Paget's disease, which was confined to a very small area. The points of particular interest were (1) the association of Paget's disease with a cloacogenic carcinoma, and (2) the dissimilarity between the pagetoid cells and the carcinoma cells. The histologic differences between the two cell types support the view that perianal Paget's disease and underlying carcinomas represent a multicentric reaction to an unknown stimulus. PMID- 6271516 TI - Mucinous adenocarcinoma developing in chronic anal fistula: report of two cases and review of the literature. AB - Mucinous adenocarcinoma developing in a chronic anal fistula is a rare tumor of the anus of which there are less than 150 reported cases. There has been some debate as to whether the fistula is the source of the tumor, or whether the fistula is the presenting feature of a slow-growing, indolent carcinoma. Two recent cases seen at our hospital are presented, along with a review of the literature and what we feel to be strong evidence that the fistula and associated anal glands are indeed the source of this unusual tumor. PMID- 6271518 TI - Marihuana and sex: a critical survey. AB - Marihuana usage is associated with a life-style that involves earlier and more frequent sexual activity. Marihuana usage does not affect human male testosterone levels significantly, but does adversely affect sperm production. Animal studies have not found consistent changes in weights of male sexual organs but have corroborated the adverse effects of cannabinoid compounds on sperm production. The biological significances of these effects on sperm production are unclear, however, since there is no evidence that human marihuana users or male animals given cannabinoid compounds are less fertile or are at risk for dominant lethal mutations. Cannabinoid compounds reliably inhibit ovulation in animals and are associated with depressed luteinizing hormone (LH) levels in both female and male animals. The decreased LH levels appear to be due to both hypothalamic and ovarian sites of action. Treatment with cannabinoid compounds is also associated with lower testosterone levels in male and lower prolactin levels in female animals. Effects on progesterone levels are inconclusive. Cannabinoid compounds do not possess estrogenic activity. Despite some consistencies in the data in virtually every study conducted with animals, there has been a basic confounding between direct drug action and secondary effects resulting from drug-induced decreases in food and water consumption and attendant weight loss. Almost all of the adverse effects of cannabinoid exposure on reproductive organs can be attributed to these secondary effects. PMID- 6271519 TI - A critical review of the safety and antiemetic efficacy of delta-9 tetrahydrocannabinol. AB - Over the past six years, both government institutions and private enterprise have expressed great interest and activity in the isolation of chemical constituents from the Cannabis sativa plant and synthesis of novel cannabinoid compounds with potential medicinal uses. Evaluation of the safety and antiemetic efficacy of delta-9-tetrahydrocannabinol, the primary psychoactive cannabinoid component of marijuana, comprises one of several very active areas of cannabinoid clinical research. The results of clinical studies of the safety and antiemetic efficacy of delta-9-tetrahydrocannabinol in patients receiving cancer chemotherapy are critically evaluated. Deficiencies in current knowledge of the clinical pharmacology of delta-9-tetrahydrocannabinol are discussed in the context of antiemetic drug evaluation. PMID- 6271517 TI - Complete regression of hepatocellular adenoma after withdrawal of oral contraceptives. AB - A 28-year-old woman who complained of mild abdominal pain was found to have a large liver tumor. Angiography and needle biopsy established the diagnosis of hepatocellular adenoma. The tumor was felt to be unresectable because of its size, and no treatment was given other than withdrawal of oral contraceptives. Subsequent hepatic scintiscans documented compete resolution of the tumor over a 12-month period. PMID- 6271520 TI - A controlled study of hepatitis core antibody (HBcAb) in primary hepatocellular carcinoma and liver cirrhosis in Kenya. PMID- 6271521 TI - Cytogenetic effects on the eastern mudminnow, Umbra pygmaea, exposed to ethyl methanesulfonate, benzo[a]pyrene, and river water. PMID- 6271522 TI - [Diagnostic and therapeutic problems in atypical carpal tunnel syndromes]. PMID- 6271523 TI - [Electrology in functional re-education]. PMID- 6271524 TI - In vitro effects of parathyroid hormone on kidney cortical slices: cAMP responses and concomitant inhibition of the Na+ gradient-dependent uptake of phosphate by brush border membrane vesicles isolated from the renal slices. AB - Mouse renal cortical slices were incubated with parathyroid hormone (30 U/ml) for 2 min. Brush border membrane vesicles isolated from the treated slices had a decreased Na+ gradient-dependent uptake of phosphate. Concomitantly, the hormone elicited the activation of adenylate cyclase, the increase in tissue level of cAMP, and the enhancement of cAMP-dependent protein kinase. PMID- 6271525 TI - Delayed puberty induced by chronic suppression of prolactin release in the female rat. AB - To study the effect of PRL deficiency on the onset of puberty, PRL release was chronically inhibited by treating immature female rats with the dopaminergic receptor agonist, bromoergocriptine (CB-154). The resulting alterations in the time of puberty onset, and in other associated parameters, such as serum levels of pituitary hormones, ovarian responsiveness to gonadotropins and ovarian hCG receptor content were then evaluated. CB-154 was provided in the drinking water from day 22 onward at the concentration of 20 and 100 micrograms/ml. The treatment resulted in almost complete suppression of serum PRL levels throughout the entire period studied (day 22 to first diestrus). In contrast, serum GH, FSH, and LH levels were not depressed. Likewise, pulsatile release of FSH was not affected and only a subtle alteration in pulsatile LH release was apparent. The onset of puberty, as determined by the age at vaginal opening, and at first diestrus after the first estrus and by the presence of corpora lutea at sacrifice (first diestrus), was markedly delayed in the hypoprolactinemic (HPO) rats. This inhibitory effect of CB-154 was completely prevented by concomitant administration of PRL. Ovarian weight was significantly decreased in HPO rats at the three ages studied (27, 32, and 36 days of age). By day 36, 50% of the control animals had already ovulated, as compared with only 9% of the HPO rats. Microscopic examination of ovaries from HPO rats revealed a retarded follicular development. In vitro ovarian progesterone response to hCG studied at day 32 and 36 of age was reduced in the HPO rats. Uterine growth was also depressed in HPO rats, the ovaries of which, when incubated in vitro, failed to show the prepubertal increase in estrogen response to hCG seen in control rats between day 32 ad 36. Aromatase activity, as measured by the in vitro release of estradiol from ovaries incubated in the presence of an excess of androgen substrate, was depressed in HPO rats. hCG receptor content in the ovaries from HPO rats (counts per min [125]hCG bound per micrograms DNA) was also lower than that of control animals at day 32 and 34 but not at day 36. However, at this later time the hCG receptor content per milligram of ovary was still significantly reduced in HPO rats. The results support the view that PRL plays an important role in the process of ovarian development that leads to the onset of puberty in the female rat and that this effect is, at least in part, exerted through a positive influence of PRL on ovarian LH receptor content. PMID- 6271526 TI - Pituitary gonadotropin-releasing hormone receptors on proestrus: effect of pentobarbital blockade of ovulation in the rat. PMID- 6271527 TI - Internalization and degradation of human chorionic gonadotropin in ovine luteal cells: kinetic studies. PMID- 6271528 TI - Antibodies to purified luteinizing hormone receptor localize the receptor at the luteal cell surface. AB - Antiserum to purified LH (hCG) receptor was raised in rabbits and used to localize the receptor with the peroxidase-antiperoxidase complex method in pseudopregnant rat ovary. The receptor was purified using immobilized antibodies to hCG as an immunoaffinity using immobilized antibodies to hCG as an immunoaffinity matrix for solubilized receptor-hCG complex. The unoccupied receptor was eluted from the column by acetic acid and used to immunize rabbits. The antibodies produced inhibited binding of [125I]iodo-hCG to pseudopregnant rat ovarian particles in a dose-dependent manner, mainly by reducing the number of available binding sites. The antireceptor serum gave a positive peroxidase staining at the luteal cell periphery on ovarian sections bearing unoccupied receptors, while no staining was seen on spleen, kidney, or liver sections. No reaction was seen on ovarian sections when the receptors were saturated with hCG in vivo, while the anti-hCG serum gave a distinct peripheral reaction in luteal cells on these sections. This fact and the idea that the antibodies produced did not bind to purified receptor-[125I]iodo-hCG complex suggest that the hormone binding site and the antibody-binding site on the receptor are very near each other or identical. The immunocytochemical findings suggest that the majority of the LH (hCG) receptors in luteal cells are located at the cell surface. PMID- 6271529 TI - Kinetic aspects of cycloheximide-induced reversal of adrenocorticotropin effects on steroidogenesis and phospholipid metabolism in rat adrenal sections in vitro. AB - We examined the rate of cycloheximide-induced reversal of ACTH effects on steroidogenesis and phospholipid metabolism in adrenal sections in vitro. In the absence of cycloheximide, ACTH treatment elicited sustained increases in steroidogenesis (approximately 3- to 5-fold) and adrenal concentrations (approximately 2-fold) of phosphatidic acid, phosphatidylinositol, and polyphosphoinositides. These increases in phospholipids were not dependent on steroidogenesis, since they were also apparent in aminoglutethimide-blocked adrenal sections. The addition of cycloheximide 30 min after ACTH treatment reversed the stimulatory effects of ACTH on steroidogenesis and phospholipids, and the rates of decay for both processes were identical, viz 28 min. This relatively slow turnover in adrenal sections in vitro contrasts with a much more rapid turnover observed previously in vivo and presently in dispersed adrenal cells in vitro; thus, it appears that an artefact of the in vitro adrenal section system stabilizes the putative labile protein. More importantly, the identical rates of decay for steroidogenesis and phospholipids suggest that the same labile protein is required for both phospholipid metabolism and steroidogenesis. These findings are in keeping with our postulate that the labile protein is required for phospholipid metabolism, which, in turn, is required for steroidogenesis. PMID- 6271530 TI - Decreased thyroidal response to thyrotropin in diabetic mice. AB - The effect of diabetes mellitus on the synthesis and secretion of thyroid hormone ws investigated in mice with streptozotocin-induced diabetes. Thyroid glands were labeled in vivo with 131I for 2 h. In control animals, TSH stimulated the synthesis of PB127I and 131I-labeled iodothyronines and simultaneously decreased the proportion of 131I-. These effects of TSH were not observed in diabetic animals but were demonstrable in diabetic animals treated with insulin. For studies of hormone secretion, labeled thyroid glands were cultured in vitro in medium containing 1 mM mononitrotyrosine. The rate of the hydrolysis of labeled thyroglobulin was measured as the proportion of 131I-labeled iodotyrosines and 131I-labeled iodothyronines recovered at the end of culture and was used as an index of thyroid secretion. TSH in vivo stimulated the rate of thyroglobulin hydrolysis for 6 h, with a peak occurring after 2 h. The diabetic mice had a diminished response to TSH, which improved on treatment with insulin. The addition of TSH and insulin to the culture medium significantly increased the rate of thyroglobulin hydrolysis in glands of diabetic mice over that resulting from the addition of dibutyryl cAMP alone. The generation of thyroidal cAMP in response to TSH was higher in diabetic mice than in controls. The rise in plasma T4 and T3 2 h after the administration of TSH was less in diabetic mice than in control mice or diabetic mice treated with insulin. Our studies, therefore, indicate that the thyroidal response to TSH is decreased in diabetes mellitus. The defect appears to be at a step beyond the generation of cAMP. PMID- 6271531 TI - The effect of vitamin D restriction and repletion on bone apposition in the rat and its dependence on parathyroid hormone. AB - The regulatory role of vitamin D in bone formation and its interaction with parathyroid hormone (PTH) were analyzed in rats in vivo. The bone apposition rate was determined by measuring the distances between tetracycline lines deposited at 48-h intervals. Vitamin D restriction was associated with a decrease in the bone apposition rate and was paralleled by the expected decrease in serum 25 hydroxyvitamin D3 levels but not by changes in 1,25-dihydroxyvitamin D3 levels. Vitamin D3 repletion restored the apposition rate to normal in parathyroid-intact animals but had no effect in parathyroidectomized animals. Vitamin D repletion of PTH-repleted parathyroidectomized animals restored the bone apposition rate. These results indicate that vitamin D or vitamin D metabolites are required for the expression of the effect of PTH on bone apposition. Conversely, vitamin D repletion only affects the bone formation rate in the presence of PTH, thus suggesting the requirement for PTH-dependent vitamin D metabolism. PMID- 6271532 TI - Triiodothyronine augments the number of membrane-bound (Na+-K+)-adenosine triphosphatase units, but does not affect the sedimentation properties of plasma membrane components. AB - We have previously demonstrated that T3 enhanced the de novo synthesis of renal cortical (Na+-K+)-dependent ATPase (NaK-ATPase) in the rat. A purified membrane fraction obtained from successive centrifugation of renal cortical crude homogenate was used in the above studies. To rule out a possible effect of T3 on plasma membrane properties, such as the sedimentation characteristic of NaK ATPase, we have presently observed T3-dependent increases in the activity and number of NaK-ATPase units in a crude homogenate of rat renal cortex. The following results were obtained which substantiate a specific effect of T3 on the membrane-bound NaK-ATPase system. 1) Compared to the hypothyroid state, 43% and 44% increases in the activity and number of NaK-ATPase units, respectively, were observed in crude renal cortical homogenate of T3-treated hypothyroid rats. 2) In comparison with the crude homogenate prepared from hypothyroid rats, 4.8- and 113 fold increases in the specific activity of NaK-ATPase were obtained in the L and J fractions, respectively. Increases of similar relative magnitude in the L and J fractions were also shown in T3-treated hypothyroid and euthyroid rats. 3) No difference in the recovery of the number of NaK-ATPase units was observed from successive steps of the purification under different thyroid states. 4) Treatment of renal cortices from hypothyroid, T3-treated hypothyroid, and euthyroid rats with deoxycholate increased to the same extent NaK-ATPase activity and phosphorylated intermediate formation. PMID- 6271533 TI - Uterine prostaglandin concentrations in sheep during late pregnancy and adrenocorticotropin-induced labor. AB - Concentrations of prostaglandin E (PGE), PGF, and 6-oxo-PGF1 alpha (the hydrolysis product of PGI2) were measured by RIA in different areas of myometrium, endometrium, cotyledons, amnion, and chorioallantoic membrane taken from chronically catheterized sheep. The animals were 130 days pregnant and had been treated for 70 h with continuous intrafetal infusions of either ACTH-(1-24) or saline. Maternal and fetal plasma samples were collected at 8-h intervals beginning 48 h before the start of infusion, and maternal plasma progesterone, estrone, and estradiol and fetal plasma cortisol concentrations were estimated by RIAs. As suggested by a significant fall in maternal plasma progesterone and rises in maternal plasma estrone and fetal plasma cortisol concentrations, sheep receiving intrafetal ACTH were in early labor at the time tissue samples were taken for PG determinations. For myometrium, analysis of variance indicated a significant effect of uterine area on PG concentrations. Concentrations of 6-oxo PGF1 alpha were elevated toward the cervical end of the myometrium in ACTH treated animals. PGF and PGE concentrations were higher at the tubal and cervical ends of the myometrium in both groups of animals. In the endometrium of treated animals, concentrations of PGE, but not PGF or 6-oxo-PGF1 alpha, were elevated significantly. For cotyledons and chorioallantois, concentrations of PGF and 6 oxo-PGF1 alpha, but not PGE, were elevated significantly in ACTH-treated animals. PG concentrations were not elevated in amnion taken from ACTH-treated animals. For both groups, the areas of highest PG concentrations were cotyledons and chorioallantois, suggesting that these may be major sites of synthesis. PMID- 6271534 TI - Effects of prostaglandins on adenosine 3',5'-monophosphate content and adenylate cyclase activity in dispersed bovine parathyroid cells. PMID- 6271535 TI - In vivo demonstration of receptors in rat liver to the amino-terminal region of parathyroid hormone. AB - The specific binding of the amino-terminal region of parathyroid hormone (PTH) to liver has been assessed by in vivo radioautography employing a biological active 125I-labeled synthetic bPTH analog ([Nle-8,18, Tyr-34]bPTH-(1-34) amide) as a probe. Two minutes after intrajugular injection of the labeled analog into rats, free hormone was separated from that bound to cells by intracardiac perfusion with buffer (30 sec), followed by perfusion-fixation with glutaraldehyde. By light and electron microscope radioautography, the distribution of silver grains over the liver sections revealed localization over the periphery of hepatocytes as well as over endothelial cells. In simultaneous control experiments, the unlabeled synthetic active fragment bPTH-(1-34) and unlabeled intact native bPTH (1-84) significantly inhibited binding of the labeled analog to liver hepatocytes but not to endothelial cells. Greater uptake of the 125I-labeled bPTH analog was found in rat liver (28% of the injected dose) than in kidney, bone, or other organs examined. In parallel experiments using purified plasmalemma fractions from hepatocytes, both bPTH-(1-34) and intact bPTH-(1-84) demonstrated a dose dependent activation of adenylate cyclase which was less than that of glucagon but greater than that of epinephrine. The combined results support the concept of the liver as a target organ for the amino-terminal biologically active region of PTH. PMID- 6271536 TI - Regulation of thyroid adenylate cyclase: guanyl nucleotide modulation of thyrotropin receptor-adenylate cyclase function. AB - The role of guanyl nucleotides in regulating the hormonal responsiveness of adenylate cyclase was studied in thyroid plasma membranes. Guanyl-5'-yl imidodiphosphate [Gpp(NH)p] alone stimulated the enzyme. At a low ATP concentration (0.2 mM), TSH alone had little or no effect, but when added with Gpp(NH)p, it resulted in a dose-dependent increase of enzyme activity. Kinetic studies revealed that Gpp(NH)p alone stimulated adenylate cyclase activity only after a 10-min lag. TSH abolished the lag, resulting in an apparent increase in activity and a lowering of the activation constant for Gpp(NH)p. GTP caused an initial increase in activity at 2 min, followed by a gradual decline below basal levels. This inhibition was not prevented by TSH. Further examination revealed that GDP caused inhibition of Gpp(NH)p-stimulated activity in a competitive manner, suggesting that conversion of GTP to GDP may be responsible for the time dependent decay seen with GTP. To study the effects of guanyl nucleotides on coupling of the TSH receptor to adenylate cyclase, plasma membranes were preactivated with saturating amounts of Gpp(NH)p and washed extensively to remove unbound Gpp(NH)p. Incubation of preactivated membranes with either Gpp(NH)p or TSH gave no further stimulation of adenylate cyclase. However, Gpp(NH)p plus TSH produced 30% more stimulation. In contrast, the addition of TSH plus GDP to preactivated membranes led to a dose-dependent decrease in activity. Furthermore, promotion of further stimulation by TSH plus Gpp(NH)p was competitively inhibited by GDP, in the same manner as untreated membranes. This suggested that the dual action of TSH, i.e, stimulation and inhibition, is governed and mediated by specific guanyl nucleotides. Analysis of guanyl nucleotides effects on the binding of [125I[iodo-TSH to membranes revealed that although some inhibition of binding exists, this effect is 1) unrelated to the concentration effect of nucleotides on adenylate cyclase, and 2) not specific for guanosine phosphates. Scatchard analysis of TSH binding in the presence of guanyl nucleotides demonstrated that even at high concentrations, GTP had no effect on the high affinity, low capacity receptor for TSH. These results suggest that GDP or Gpp(NH)p plays no role in modulating TSH-receptor interaction. Thus, guanyl nucleotide regulation of TSH action appears to be limited to adenylate cyclase. PMID- 6271537 TI - The ontogeny of the thyrotropin-thyroid axis in early bovine embryos. AB - T4 and T3 formation and their response to TSH, cAMP, and prostaglandin E2 (PGE2) stimulation were studied by RIA in cultured bovine fetal thyroids from 130 embryos of 1.2-25.0 cm crown-rump length (CRL). T4 and T3 were found in all of the freshly isolated glands studied, and their concentrations increased significantly (P less than 0.05) during a 24-h incubation of glands from fetuses with a CRL greater than 8.0 cm. The release of T4 (nanograms per mg tissue), but not of T3, increased consistently with CRL (r = 0.64; P less than 0.05). The addition of TSH (0.5 mU) to the culture medium induced a 2- to 3-fold increase in the secretion of T4, but not of T3, by glands of fetuses with a CRL of 3.0-25.0 cm (P less than 0.05). Dibutyryl cAMP (10(-4) M) and PGE2 (10(-4) M) had comparable effects. A combination of TSH and theophylline (1 mM) or of TSH and dibutyryl cAMP significantly enhanced the T4 released by cultured tissue into the medium (P less than 0.05) over that induced by either agonist, but the combined effect was not fully additive. The total PGE2 in the tissue and medium was not changed by the addition of 0.5 mU TSH, and indomethacin had no effect on TSH induced T4 secretion. The data show that thyroids of fetuses that have reached a CRL of 3.0 cm have the enzymatic capacity to produce both T4 and T3, and T4 is the dominant product formed. While exogenous PGE2 at high concentrations stimulates fetal T4 secretion, it does not mediate the actions of TSH and cAMP on the fetal thyroid. PMID- 6271538 TI - Biosynthesis of proteoglycans by rat granuloma cells cultured in vitro: modulation by gonadotropins, steroid hormones, prostaglandins, and a cyclic nucleotide. AB - Rat ovarian granulosa cells were isolated from immature female rats 48 h after stimulation with 5 IU PMS gonadotropin and then maintained in culture. The effects of ovine (o)FSH, oLH, hCG, testosterone, 17 beta-estradiol, progesterone, prostaglandins E1, E2, F1 alpha, F2 alpha, N,O'-dibutyryl cAMP, and theophylline on proteoglycan synthesis by the granulosa cells in vitro were examined using [35S]sulfate as a precursor. oFSH, oLH, hCG, prostaglandins E1 and E2, N,O' dibutyryl cAMP, theophylline and testosterone stimulated the production of 35S labeled proteoglycans compared to control cultured. 17 beta-Estradiol, progesterone, and the prostaglandins F1 alpha, and F2 alpha, showed no stimulation. Two major species of proteoglycan are synthesized and secreted into the medium: 1) a population with large hydrodynamic size (Kav = 0.30 on Sepharose CL-2B); and 2) a population with relatively small size (Kav = 0.60 on Sepharose CL-2B). The stimulatory effect of the hormones was accounted for entirely by a net increase in synthesis of the smaller proteoglycan population. The hormones exert their stimulatory effects on proteoglycan synthesis at concentrations similar to those required for other in vitro biological responses of granulosa cells. PMID- 6271539 TI - Postnatal maturation of gonadotropes in the male rat pituitary. AB - The postnatal maturation of gonadotropes was studied with the use of electron microscopic-immunocytochemical strains for the beta-chains of LH and FSH on serially sectioned fields. A third serially sectioned field was stained with antiserum to ACTH-(17-39). Morphometric studies on semithin and ultrathin plastic sections stained for LH and FSH showed a 200-300% increase in the percentages of LH and FSH cells during the first week of postnatal life. In the 1- to 2-week-old rats, the percentage of gonadotropes in the population was greater than that in the adult. Analysis of the hormone content of serially sectioned gonadotropes showed that storage patterns similar to those in the adult were reached by the seventh day of life. In the neonatal group, 66% of the serially sectioned cells contained both FSH and LH, 20% contained only LH, and 14% contained only FSH. In the 7- to 15-day-old rats, the percentages of gonadotropes containing both hormones increased to 79% while 10-11% of the cells contained only LH or FSH. This is similar to the findings in the adult rat population (75% LH and FSH cells, 14.2% LH cells, and 11% FSH cells). The fields stained for ACTH showed that 35-80% of the gonadotropes contained ACTH-like immunoreactivity during neonatal development, with the highest values seen in the 7- to 15-day-old rats. In the normal adult rats, 2-10% of the gonadotropes contain ACTH activity. Cells containing only ACTH were also seen in all age groups examined. The morphological analysis showed that immature gonadotropic cells contained scattered storage granules, arranged either at the cell periphery or concentrated near the nucleus. They were small, irregularly shaped, and often exhibited a high nucleo cytoplasmic ratio. They were difficult to distinguish from corticotropes. More mature cell types were found after 1 week of age. These were ovoid and contained numerous large and small granules, features similar to the adult type I cell. The granules, however, were more irregularly shaped (pleomorphic). Immature forms were observed in the gonadotrope population as late as 15 days of age. Most gonadotropes from the 20- to 25-day-old rats resembled those in the adult population. Our findings support the physiological studies which show that the first week of life is an important time for functional and morphological maturation of the gonadotrope population. They also support the finding that immature gonadotropes are larger, more numerous, and more heterogeneous than those in the adult male rat population. Finally, we propose that a subpopulation of gonadotropes may serve as a stem cell or may be involved in the development of adrenal-gonadal interactions. PMID- 6271540 TI - Influence of vitamin D on parathyroid hormone-induced adenosine 3',5' monophosphate production by bone cells isolated from rat calvariae. PMID- 6271541 TI - Characterization of cholecystokinin receptors on rat pancreatic membranes. PMID- 6271542 TI - Opioid regulation of prolactin secretion: evidence for a specific role of beta endorphin. AB - Previous studies have shown that exogenously administered opioids, including beta endorphin, stimulate prolactin release. The fact that naloxone has been shown to lower baseline and stress-induced serum prolactin in rats suggests that endogenous opioids may participate in prolactin regulation, but does not specify which opioid is involved. In a previous study we showed that intravenously administered anti-beta-endorphin antiserum had no effect on either baseline or stress-induced prolactin release in rats. In the present study we have repeated our earlier experiments, but have given the antiserum into the cerebral ventricles rather than intravenously. Significant lowering of baseline serum prolactin, to 56.6% +/- 10.6% (SEM) that of controls (p less than 0.005), was noted. Also noted was blunting of the stress-induced prolactin rise, to 69.2% +/- 6.7% that of controls (p less than 0.002). These results indicate that beta endorphin is specifically involved in both the tonic and stress-mediated release of prolactin. Because the magnitude of prolactin lowering with antiserum was at least as great as what we had earlier observed with naloxone, they suggest that beta-endorphin is the major and possibly the sole opioid involved in prolactin regulation. They also indicate that the previous lack of effect of anti-beta endorphin antiserum on serum prolactin was due to its failure, after intravenous administration, to achieve effective concentration at critical controlling sites within the brain. PMID- 6271543 TI - Serum lipoproteins increase testosterone production in hCG-desensitized Leydig cells. AB - The effects of high density and low density lipoproteins (HDL and LDL) on testosterone synthesis by Leydig cells from rats treated three days earlier with 50 IU human chorionic gonadotropin (hCG) or from saline-injected control rats were investigated. HDL caused a significant, dose-dependent increase in both basal and hCG-stimulated testosterone production by Leydig cells from hCG-treated rats, but had no marked effect on Leydig cells from control rats. The serum testosterone concentration of hCG-treated rats was three times higher than in controls. Basal testosterone production by Leydig cells from hCG-treated rats was elevated two-fold in comparison to controls, while hCG-stimulated testosterone production was significantly lower than in controls. Both basal and hCG stimulated testosterone production by Leydig cells from hCG-treated rats were significantly increased by the addition of HDL or LDL. Furthermore, HDL restored the hCG-stimulated testosterone production to the maximal amount produced by hCG stimulated Leydig cells from control animals. When equivalent amounts of HDL or LDL cholesterol (140 microgram/ml) were added to the incubation, HDL caused a greater increase in testosterone production than did LDL. These results suggest that hCG-treatment, in vivo, caused a prolonged stimulation of the Leydig cells which resulted in depletion of the cholesterol available for steroidogenesis, since the addition of cholesterol in the form of HDL or LDL caused an increase in the testosterone production of these Leydig cells. PMID- 6271544 TI - The biosynthesis, intracellular processing, and secretion of parathormone. PMID- 6271545 TI - Relationships between circulating and intracellular thyroid hormones: physiological and clinical implications. PMID- 6271547 TI - Gonadotropin-releasing hormone receptors: characterization, physiological regulation, and relationship to reproductive function. PMID- 6271546 TI - Gonadotropin-releasing hormone action in the pituitary: a three step mechanism. PMID- 6271548 TI - Coagulation of human plasma by asbestos fibers. PMID- 6271549 TI - Manifestations of cellular immunity in the rat after prolonged asbestos inhalation. II. Alveolar macrophage-induced splenic lymphocyte proliferation. PMID- 6271550 TI - Biological reactivity of calcium silicate composites--in vitro studies. PMID- 6271551 TI - Oxygen activation and tetrapyrroles. PMID- 6271552 TI - Control of synthesis of wall teichoic acid in phosphate-starved cultures of Bacillus subtilis W23. AB - CDP-glycerol pyrophosphorylase, CDP-ribitol pyrophosphorylase and poly(ribitol phosphate) synthetase activities have been measured in cultures of Bacillus subtilis W23 as they became phosphate-starved either in batch culture or during changeover from potassium limitation to phosphate limitation in a chemostat. The results indicated that repression of synthesis of all three enzymes occurred at the onset of phosphate starvation and that this was accompanied by inhibition of inactivation of CDP-glycerol pyrophosphorylase and poly(ribitol phosphate) synthetase. These results show that the initial response to phosphate starvation involves more than inhibition of one enzyme as proposed by Glaser and Loewy [Glaser L. and Loewy, A. (1979) J. Biol. Chem. 254, 2184-2186]. Synthesis of both linkage unit and poly(ribitol phosphate) are inhibited independently. PMID- 6271553 TI - Biochemical characterization of skin fibroblasts derived from WHHL-rabbit, a notable animal model for familial hypercholesterolemia. PMID- 6271554 TI - The origin of metastatic heterogeneity in tumors. PMID- 6271555 TI - Variant translocation in a non endemic case of Burkitt's lymphoma: t (8;22) in an Epstein--Barr virus negative tumour and in a derived cell line. PMID- 6271556 TI - Activation of the glycogenolytic cascade in human polymorphonuclear leucocytes by different phagocytic stimuli. AB - Human polymorphonuclear leucocytes were found to respond to activation by immunoglobulin opsonized latex particles and to complement opsonized zymosan particles with a rapid transient increase in cAMP concentration, dissociation of the cAMP dependent protein kinase, activation of glycogen phosphorylase and glycogen break down. However, since phosphorylase kinase was not activated, the activation of phosphorylase is believed to be secondary to non-covalent activation of phosphorylase kinase by Ca2+. Activation by the soluble stimulator phorbol myristate acetate resulted in activation of phosphorylase and glycogen break down, whereas no changes in cAMP concentration, protein kinase activity, or phosphorylase kinase activity were observed. The activation of phosphorylase is ascribed to an increase in cytosolic Ca2+ concentration. The response to stimulation by zymosan was strongly inhibited by ethylene glycol-bis-(beta aminoethyl ether)-N,N1-tetraacetic acid, which did not affect stimulation by either latex particles or phorbol myristate acetate. The same differential effect of ethylene glycol-bis(beta-aminoethyl ether)-N,N1-tetraacetic acid was observed when the response of the cells was measured as increase in oxygen consumption and activation of the hexose monophosphate shunt. PMID- 6271557 TI - Cytotoxic T cell recognition of Epstein-Barr virus-infected B cells. II. Blocking studies with monoclonal antibodies to HLA determinants. AB - Monoclonal antibodies specifically binding common determinants on all HLA-A, B anc C antigen molecules blocked the lysis of EB virus-transformed target cells by EB virus-specific cytotoxic T cells reactivated in vitro. Blocking was mediated through binding of the antibodies to the target rather than to the effector cells and was maximal (75 to 85% inhibition of lysis) at saturating antibody concentrations. A similar blocking effect was also shown by a monoclonal antibody binding to beta 2-microglobulin, a molecule physically associated with HLA-A, B and C antigens on the target cell surface, but not by a monoclonal antibody binding to the non-HLA-associated leukocyte-common antigen. Saturating concentrations of monoclonal antibodies specific for common determinants on all HLA-DRw antigen molecules either had no effect at all upon EB virus-specific T cell cytotoxicity or caused a slight, but nonspecific, inhibition. The results demonstrate unequivocally that HLA-A, B and C antigens on the target cell surface are indeed the polymorphic elements which impose genetic restriction upon EB virus-specific cytotoxic T cell function. PMID- 6271558 TI - Binding and metabolism studies with [3H](D-Ala2,Leu5)enkephalinamide and [3H](D Ala2,Pro5)enkephalinamide: evidence for a selective interaction of (D Ala2,Pro5)enkephalinamide with mu-receptors. AB - We have compared the binding characteristics of [3H](D-Ala2,Leu5)enkephalinamide and [3H](D-Ala2,Pro5)enkephalinamide on a washed homogenate from mouse brain. The maximum number of binding sites for [3H](D-Ala2,Leu5)enkephalinamide was twice that obtained with [3H](D-Ala2,Pro5)enkephalinamide. Hill slopes of the displacement curves for opiates and (D-Ala2,Pro5)enkephalinamide obtained against [3H](D-Ala2,Leu5)enkephalinamide were considerably lower than that of (D Ala2,Met5)enkephalinamide and (D-Ala2,Leu5)enkephalinamide. In contrast, Hill slopes for morphine, ketocyclazocine and (D-Ala2,Leu5)enkephalinamide were similar for the displacement of [3H](D-Ala2,Pro5)enkephalinamide. These results are discussed and interpreted in terms of a selective interaction for (D Ala2,Pro5)enkephalinamide with mu-receptors and a similar affinity of (D Ala2,Leu5)enkephalinamide for mu- and delta-receptors. Since metabolism studies did not show any difference in the degradation rate of these two enkephalin analogs, the discrepancy between in vitro and in vivo activity cannot result from the catabolism processes. Another possible explanation is proposed. PMID- 6271559 TI - Alpha 2-adrenoreceptors on human platelets: selective labelling by [3H]clonidine and [3H]yohimbine and competitive inhibition by antidepressant drugs. AB - [3H]Clonidine and [3H]yohimbine have been used to characterize alpha 2 adrenoreceptors on human platelets. At 25 degrees C binding was rapid (t 1/2 of association, 1.8 and 2.7 min) and reversible (t 1/2 of dissociation, 0.5 and 8.2 min). The binding sites for [3H]clonidine and ]3H]yohimbine showed the specificity required for an alph 2-adrenoreceptor. The rank order of potency of inhibitors of [3h[clonidine binding was clonidine greater than yohimbine greater than phenylephrine greater than prazosin and of [3H]yohimbine binding was yohimbine greater than clonidine greater than phenylephrine greater than prazosin. Scatchard analysis of [3H]yohimbine binding indicated the existence of a single population of noninteracting sites (KD = 3.0 nM; Bmax = 188 fmol/mg protein). The high-affinity binding of [3H-clonidine had a lower affinity and a lower number of sites (KD = 5.0 nM; Bmax = 35 fmol/mg protein). [3H]Clonidine binding also showed evidence of a second site of much lower affinity and greater number (KD = 18.6 nM; Bmax = 77 fmol/mg protein) in 40% of the normal population. In vitro, antidepressant drugs competed with [3H]clonidine and [3H]yohimbine for the platelet alpha 2-adrenoreceptor. The rank order of potency of inhibitors of [3H]clonidine binding was mianserin greater than amitriptyline greater than iprindole greater than desipramine and of [3H]yohimbine binding was mianserin greater than amitriptyline greater than desipramine greater than iprindole. The inhibition constants (Ki) of adrenergic drugs and of various antidepressant drugs in competing with [3H]clonidine were correlated with the inhibition constants of these drugs in competing with [3H]yohimbine (r = 0.970; P less than 0.001) which suggests that both radioligands labelled the same alpha 2-adrenoreceptor on the human platelet. The inhibition of binding induced by all antidepressant drugs was competitive. In contrast, long-term administration of tricyclic antidepressant drugs to patients was recently found to be associated with a decrease in the number of binding sites for [3H]clonidine on platelet membranes. The present results indicate that both [3H]clonidine and [3H]yohimbine are useful tools for the quantification of alpha 2-adrenoreceptors on blood platelets and suggests that the specific binding of radiolabelled alpha 2-adrenoreceptor ligands to human platelet membranes might be used to monitor changes in alpha 2 adrenoreceptors during tricyclic antidepressant drug treatment. PMID- 6271560 TI - Strain differences in opiate receptors in mouse brain. AB - Various opiate ligands were bound to brain membranes of mice of the Recombinant Inbred System. The specific binding of low levels of [3H]naloxone, [3H]dihydromorphine and [3H]ethylketocyclazocine was disturbed in a similar fashion among the inbred strains, and in a pattern different from that observed for [3H](D-Ala2,D-Leu5)-enkephalin. The results indicate that the inbred strains differ in mu- and delta-type binding and support the concept of multiple opiate receptors in mouse brain. PMID- 6271561 TI - Suppressive effect of cholecystokinin and its related peptides on beta-endorphin induced catalepsy in rats. AB - The effects of the C-terminal octapeptide of cholecystokinin (CCK-8) and its related peptides on the onset and duration of beta-endorphin-induced catalepsy on injection of the peptides into the lateral ventricle were investigated in male rats. The onset of catalepsy was delayed to some extent by nonsulfated CCK-8 and CCK-7 but CCK-8 and caerulein were ineffective. Naltrexone and caerulein significantly shortened the duration of catalepsy, but CCKs were less effective to shorten it. Pentagastrin had no effect on either parameter. PMID- 6271562 TI - Captopril impairs the vascular smooth muscle contraction mediated by postsynaptic alpha 2-adrenoceptors in the pithed rat. PMID- 6271563 TI - Correlations between sister chromatid exchange frequencies and replicon sizes. A model for the mechanism of SCE production. PMID- 6271564 TI - Glycopeptides prepared from mouse cerebrum inhibit protein synthesis and cell division in baby hamster kidney cells, but not in their polyoma virus-transformed analogs. PMID- 6271565 TI - Presence of double stranded regions of viral RNA in infected cells. AB - Ribonuclease treatment of rhinovirus-infected human embryo lung cells after cell disruption reveals that double stranded RNA is present in the preparation before nucleic acids are extracted with phenol. This shows that the hydrogen bonding between complementary molecules of viral RNA which occurs in infected cells is not a result of the extraction of RNA with phenol. PMID- 6271566 TI - Increased urinary excretion of cyclic nucleotides in X-linked hypophosphatemic (Hyp) mice. AB - Hyp mice, a model for human X-linked hypophosphatemia, had elevated urinary cyclic AMP, cyclic GMP, and magnesium excretion compared to normal mice. The data suggest a renal origin of the urinary cyclic nucleotides. No significant differences in plasma cyclic AMP and cyclic GMP were observed between genotypes. PMID- 6271567 TI - Effect of taurine on mitogen response of human lymphocytes. AB - Taurine selectively inhibits the phytohemagglutinin-stimulated incorporation of 3H-thymidine by human cultured lymphocytes (50% inhibition by 12.5 mM taurine). Decreasing effects of taurine on Na-K ATPase activity calcium accumulation by lymphocytes might be responsible for its action on cell proliferation. PMID- 6271568 TI - [Substances with antiviral activity. XIX. Amidinohydrazones of formylesters and formylthioesters with aromatic N-heterocyclic substrates]. AB - Some formylthioester and formylester amidinohydrazones with aromatic N heterocyclic moieties, were synthesized and screened in vitro against the MP mutant of herpes simplex virus type 1 [HSV-1 (MP)] and parainfluenza type 3 virus, HA-1/CR-8 strain. The amidinohydrazone of 1-phenyl-4-formyl-3 (ethylthio)carbonylpyrrole (I) was the most active compound. PMID- 6271569 TI - Relationship between the binding of dicyclohexylcarbodiimide and the inhibition of H+-translocation in submitochondrial particles. PMID- 6271570 TI - Atractyloside and bongkrekic acid sites in the mitochondrial ADP/ATP carrier protein. An appraisal of their unicity by chemical modifications. PMID- 6271571 TI - Formation of two-dimensional crystals in pure membrane-bound Na+,K+-ATPase. PMID- 6271572 TI - Phosphorylation of the 100 000 Mr Ca2+-transport ATPase by Ca2+ or cyclic AMP dependent and -independent protein kinases. PMID- 6271573 TI - Gonadotropin-induced phosphorylation of endogenous proteins in the Leydig cell. PMID- 6271574 TI - An enzymatic approach for localization of oligodeoxyribonucleotide binding sites on RNA. Application to studying rRNA topography. PMID- 6271575 TI - Involvement of tyrosyl residues in the binding of benzodiazepines to their brain receptors. PMID- 6271576 TI - A K+-selective conductance sensitive to cholinergic antagonists obtained by the fusion of axonal membrane vesicles to planar bilayers. PMID- 6271577 TI - Characterization of bovine ceruloplasmin. PMID- 6271578 TI - Selective extraction of biologically active F-glycoprotein from dithiothreitol reduced Sendai virus particles. PMID- 6271579 TI - Relaxed specificity of endonuclease BamH1 as determined by identification or recognition sites in SV 40 and pBR 322 DNAs. PMID- 6271580 TI - Variations in luteinizing hormone-releasing hormone receptors in pituitary cells from immature and mature cycling female rats. PMID- 6271581 TI - The energy-dependent unmasking of -SH groups in the mitochondrial ADP/ATP carrier, and its prevention by nigericin. PMID- 6271582 TI - Effects of arachidonate on cultured pig thyroid cells and their stimulation by thyrotropin. PMID- 6271583 TI - 19F-nuclear magnetic relaxation by superoxide dismutase as an enzymic method for the detection of superoxide ion. PMID- 6271585 TI - Restriction endonuclease BamH1 interaction with a synthetic duplex containing half-size recognition sequences. PMID- 6271584 TI - Effect of indomethacin on the binding of the chemotactic peptide formyl-Met-Leu Phe on human polymorphonuclear leukocytes. PMID- 6271586 TI - Binding of LP(a) to the low density lipoprotein receptor of human fibroblasts. PMID- 6271587 TI - Pyruvate kinase phosphatase. PMID- 6271588 TI - Determination of the free-energy difference of the adenine nucleotide translocator reaction in rat-liver mitochondria using intra- and extramitochondrial ATP-utilizing reactions. PMID- 6271589 TI - Protein A24 lyase is an isopeptidase. PMID- 6271590 TI - Selective disorganisation of biochemical function in B cells of islets of Langerhans infected by EMC-M virus in tissue culture. PMID- 6271591 TI - The fate of VPg during in vitro translation of poliovirus RNA. PMID- 6271592 TI - The coordination environment of mitochondrial cytochromes b. PMID- 6271593 TI - Binding of [3H]ethyl-beta-carboline-3-carboxylate to brain benzodiazepine receptors: effect of drugs and anions. PMID- 6271594 TI - Incorporation of ATP synthetase into long-term stable liposomes of a polymerizable synthetic sulfolipid. PMID- 6271595 TI - Oudemansin, strobilurin A, strobilurin B and myxothiazol: new inhibitors of the bc1 segment of the respiratory chain with an E-beta-methoxyacrylate system as common structural element. PMID- 6271596 TI - Accessibility of metaphase chromosomes from Chinese hamster ovary cells to DNase II. PMID- 6271597 TI - Heterogeneity and properties of opiate receptors. PMID- 6271598 TI - Effects of opioid peptides on thermoregulation. PMID- 6271599 TI - ACTH and alpha-melanotropin in central temperature control. AB - Adrenocorticotropin (ACTH) and alpha-melanotropin (alpha-MSH) occur in brain tissue known to be important to temperature control. These peptides cause hypothermia if they are injected centrally in sufficient doses, but they do not act on the central set point of temperature control. Instead they appear to inhibit central pathways for heat conservation and production. In addition to their hypothermic capability, these peptides are antipyretic when given centrally in doses that have no effect on normal body temperature. ACTH has previously been associated with fever reduction in both clinical and experimental studies, and it may be that endogenous central ACTH is important for limitation of maximal fever. The hypothermic and antipyretic effects of ACTH do not depend on stimulation of the adrenal cortex because they are also observed in adrenalectomized rabbits. Nor is the antipyretic effect limited to the rabbit inasmuch as a comparable effect has been demonstrated in the squirrel monkey. The two peptides may be involved in central mediation of normal thermoregulation and fever, perhaps limiting the febrile response and other rises in body temperature by acting as neurotransmitters or neuromodulators in central thermoregulatory pathways. PMID- 6271600 TI - [Effect of acetylcholine and acetylcholinesterase inhibitors on rhythmic activity of amphibian lymphatic center motoneurons]. PMID- 6271601 TI - [Facilitation of transmission at neuromuscular synapses after "collisions" between endplate potentials and muscle action potentials induced by intracellular stimulation]. PMID- 6271602 TI - Genetic control of Rous sarcoma regression in inbred lines of chickens. AB - The fate of tumours induced by PR-RSV-C was investigated in highly inbred chicken lines congenic at the B complex, the major histocompatibility complex of the chicken. The results demonstrated that Rous sarcoma regression in congenic CB and CC lines was controlled by a gene within the B region of the chromosome. Another congenic pair of the WA and WB lines also differed in the ability to regress RSV induced tumours and a gene within the B complex also appeared to have a significant role in the regression of Rous sarcomas. PMID- 6271603 TI - Effects of cyclic nucleotides on growth activity of V79-1A cells. AB - We studied the effects of cyclic nucleotides and theophylline on growth activity of Chinese hamster V79-1A cells after extracellular application. Cyclic AMP inhibited the growth activity of V79-1A cells only at extracellular concentrations higher than 10-3 M and dibutyryl cAMP only at extracellular concentrations higher than 10-4 M. Theophylline inhibited V79-1A cell growth at concentrations higher than 10-4M. We demonstrated that cAMP or dibutyryl cAMP applied together with theophylline exerted a synergistic inhibitory effect on V79 1A cell growth. The synergistic inhibitory effect of cAMP and cGMP was found, too. Cyclic AMP at extracellular concentration of 10-10M was able to stimulate the growth of V79-1A cells slightly. Cyclic CMP slightly stimulated the proliferation of V79-1A cells at a concentration of 10-8 M. Cyclic GMP applied alone was not demonstrated as V79-1A cell growth stimulator. However, 10-6 M cGMP applied together with 10-5 M theophylline exerted a small but recognizable stimulatory effect. PMID- 6271604 TI - Location of the gene responsible for Rous sarcoma regression in the B-F region of the B complex (MHC) of the chicken. AB - We studied the fate of tumours induced by PR-RSV-C and B77-C in inbred chicken lines CB, CC and CB.R1. Rous sarcomas regressed in the CB and CB.R1 lines which are identical in the B-F region of the B complex. In contrast, progressive tumour growth was observed in the CC line which differed in the B-F region from the CB and CB.R1 lines and was identical in the B-G region with the CB.R1 line. These results suggest that the gene responsible for Rous sarcoma regression is located in the B-F region of the B complex. PMID- 6271605 TI - Effect of ampicillin or amoxycillin administration on plasma and urinary estrogen levels during normal pregnancy. AB - Urinary estrogen excretion and plasma levels of conjugated and unconjugated 17 beta-estradiol and estriol were measured in 10 healthy, pregnant women before, during and after a 5-day course of orally administered ampicillin (2 g/day) or the same dose of amoxycillin. On the second and third day of ampicillin treatment the plasma level of conjugated estriol and the urinary estrogen level were lower than pretreatment values, but then returned to basal values despite continuing treatment. Plasma concentrations of conjugated and unconjugated 17 beta-estradiol and of unconjugated estriol did not show any significant change during treatment. Hormone changes in women on amoxycillin were qualitatively similar to but smaller and less consistent than those induced by ampicillin. The results indicate (1) that the use of amoxycillin instead of ampicillin in pregnancies that require estrogen monitoring of feto-placental function, has little clinical advantage, and (2) that the adverse effect of therapeutic doses of ampicillin (and amoxycillin) on estrogen levels during pregnancy may be less important than is commonly assumed. PMID- 6271606 TI - A sensitive and specific binding assay for cyclic GMP-dependent and cyclic AMP dependent protein kinases in rat liver. PMID- 6271607 TI - The role of Zn2+ in pyridoxal phosphate mediated regulation of glutamic acid decarboxylase in brain. PMID- 6271608 TI - Metabolism of exogenous uridine 5'-triphosphate, adenosine 5'-triphosphate and pyrophosphate by alkylsulfatase-producing bacteria. PMID- 6271609 TI - Studies on superoxide dismutase from cod (Gadus morhua) liver. PMID- 6271610 TI - Detection and properties of collagenase in ovarian follicle wall of domestic fowl (Gallus domesticus). PMID- 6271611 TI - Adrenergic agents as possible regulators of liver regeneration. PMID- 6271612 TI - Dissociation kinetics of the thyrotropin-receptor complex. Characterization of a slowly dissociable component. AB - The kinetics of the dissociation of thyrotropin (TSH) from human thyroid plasma membranes were studied in an attempt to further understand the molecular dynamics of the TSH--receptor interaction. Dissociation of bound [125I]TSH from thyroid plasma membranes was a biphasic process consisting of rapidly and slowly dissociable components, RDC and SDC, respectively. The dilution induced dissociation of bound [125I]TSH was enhanced by the addition of excess TSH (DEC). DEC was proportional to the dose of unlabeled TSH and its magnitude increased linearly with temperature. These results are in contrast to those reported for the kinetics of [125I]insulin dissociation. The functional significance of DEC remains largely unexplained. It was found that the fraction of SDC was dependent upon time of association in a temperature-dependent and apparently saturable process. It could not be attributed to alterations in the electrophoretic, immunologic or binding properties of [125I]TSH. Furthermore, no correlation was observed between generation of SDC and change in the Scatchard profile of TSH binding, in contrast to studies on growth hormone. These data suggest that, like some other polypeptide hormones, binding of TSH to its receptor does not proceed according to laws describing simple, rapidly reversible, bimolecular reactions. Furthermore, bound TSH undergoes a receptor-mediated conversion from a rapidly to a slowly dissociable state with time of incubation. PMID- 6271613 TI - Taste aversion following backward conditioning procedures in preweanling and adult rats. AB - Laboratory rats, 18 and 90 days old, received an intraperitoneal injection (2% body weight) of .15M lithium chloride or .9% saline 10 or 30 min before 15-min access to 12% sucrose. Additional control groups received LiCl injection followed by tap water access. Testing with a 2-bottle choice procedure revealed reliable aversion effects for both age groups at each toxicosis-flavor interval. Adult rats showed reliably greater persistence of aversion following training with the 10- than with the 30-min interval. Rat pups showed no reliable differences in aversion across training intervals. Reliably greater aversion effects occurred for adults than for pups following training at the 10-min interval. Following training at the 30-min interval a similar reliable age effect occurred on Test Trial 1; but from Trial 2 onward the magnitude of aversion was similar for pups and adult rats. PMID- 6271614 TI - Regulation of visual attention in offspring of female monkeys treated chronically with delta 9-tetrahydrocannabinol. AB - Visual attention was studied in a group of rhesus monkey infants whose mothers received daily oral treatment with low levels of delta 9-tetrahydrocannabinol (THC, 2,4 mg/kg/day) prior to and during pregnancy and throughout lactation (3.5 postnatal months). Attention was measured at 1 and 2 years of age in a standardized test situation in which animals looked at projected slides. In comparison with controls (offspring of untreated mothers), THC offspring directed more attention at slides on the 1st trial of a session. The THC offspring also engaged in relatively longer individual periods of attention on the 1st vs 2nd trials of the session and during the 1st vs 2nd session in which a given slide was presented. Further experiments varying novelty and complexity of visual stimuli suggested that changes in visual attention of THC offspring can be characterized as a failure to limit the response to novel stimuli. PMID- 6271615 TI - The endothelial cells of large and small blood vessels. AB - Recent progress on the isolation and culture of capillary endothelial cells has allowed comparison of the biochemical and physiologic properties of endothelial cells from large and small blood vessels. These cells share certain common features including monolayer formation, production of Factor VIII antigen, production of prostacyclin, and presence of Weibel-Palade bodies. Endothelial cells from capillaries, however, differ from the endothelial cells of large arteries and veins in their nutritional requirements and in their responses to growth and migration stimuli. The differences found between endothelial cells from large and small blood vessels underscore the importance of using cells derived from vessels of appropriate size when studying the macro- and microangiopathies associated with diabetes. PMID- 6271616 TI - Herpsevirus-induced atherosclerosis. PMID- 6271617 TI - Regulation of glucose metabolism in pancreatic islets. AB - We evaluated the possible role of islet glucokinase in controlling the rate of islet glucose metabolism, and thereby the rate of glucose-induced insulin release. The activities of glucokinase, hexokinase, P-fructokinase, and glyceraldehyde-P dehydrogenase were quantitated in sonicated or isotonically homogenized islet preparations using pyridine nucleotide-dependent fluorometric assays. In sonicates, about 1/4 of the islet glucose phosphorylating activity was due to an enzyme with kinetic properties similar to glucokinase; 3/4 of the activity was due to hexokinase. The procedure for determining islet glucokinase activity was improved by centrifuging isotonic islet homogenates at 12,000 x g. The supernatant fraction was enriched for glucokinase. About 1/2 of the glucose phosphorylating activity in this fraction was due to glucokinase and 1/2 was due to hexokinase. The glucokinase activity in islet homogenates was !23 of the activity of hexokinase, 1/40 of the activity of P-fructokinase, and 1/400 of the activity of glyceraldehyde-P dehydrogenase. Detailed concentration dependency curves of glucose and mannose utilization were also obtained with intact isolated pancreatic rat islets. Glucose and mannose usage in islets was governed by two superimposed hyperbolic systems differing in Km and Vmax. A high Km system (Km for glucose 11 mM and for mannose 21 mM) predominated. A low Km system (Km for glucose 215 and for mannose 530 microM) contributed about 15% to the total activity. The available data with intact islets could be rationalized by the existence of two distinct hexose phosphorylating enzymes with differing capacities and kinetic properties. These enzymes, tentatively identified as glucokinase and hexokinase, could coexist in the same cell or could be distributed among different cell types. The possible physiologic significance of these results is discussed, emphasizing the idea of dual control of glycolysis and insulin release by glucokinase and hexokinase. An earlier proposal that glucokinase serves as glucoreceptor of beta-cells [J. Biol. Chem. 243:2730 (1968)] is greatly strengthened by the present studies. PMID- 6271618 TI - Effect of sucrose overfeeding on Na,K-ATPase-mediated 86Rb uptake in normal and ob/ob mice. AB - When normal mice have their usual chow diets supplemented by free access to a solution of 10% sucrose, their caloric intake increases by about 30%. We have used this model to explore the effects of sucrose overfeeding on Na,K-ATPase mediated cation transport. After 5 days of sucrose supplementation, Na,K-ATPase mediated K uptake is increased by 88% in liver slices and 26% in intact soleus muscles of these animals. Ob/ob mice are hyperphagic on an ad libitum chow diet, and they display even greater increments in caloric intake than do thin controls when similarly allowed access to sucrose. Despite hyperphagia while on a chow diet, Na,K-ATPase-mediated K uptake by liver slices of ob/ob mice is not significantly different from that of their thin littermates. In addition, Na,K ATPase-mediated K uptake into liver slices of ob/ob mice does not significantly increase while on sucrose supplements. These findings demonstrate the influence of dietary factors on Na,K-ATPase-mediated ion transport in liver and muscle of normal mice, and suggest that ob/ob mice may have an impairment in such dietary control. These observations suggest an important role for nutritional factors in the overall regulation of monovalent cation transport, and may also have relevance for our understanding of the cellular mechanisms of dietary thermogenesis. PMID- 6271619 TI - Simultaneous inhibition of insulin and somatomedin-C binding to cultured IM-9 lymphocytes by naturally occurring antireceptor antibodies. AB - A 53-y4-old male patient with insulin-resistant diabetes was found to have circulating inhibitors of both insulin and somatomedin-C binding. Serum obtained from the patient at the time of initial presentation inhibited 50% of both 125I insulin and 125I-SM-C binding to IM-9 lymphocytes at dilutions of 1:150. Spontaneous improvement in the patient's diabetic state was associated with a simultaneous and equal decrease in the serum inhibitory titers for both radioligands. Scatchard analysis indicated that the observed serum-induced decrease in both insulin and SM-C binding was due to decreased receptor affinity, with no alteration in receptor number. The serum inhibitors of both insulin and SM-C binding were precipitated equally by Staph-A and also by 40% ammonium sulfate, suggesting they were immunoglobulins. The observation of naturally occurring autoantibodies against both the insulin and SM-C receptors suggests a structural homology between the two receptors. PMID- 6271620 TI - Effects of a converting enzyme inhibitor and angiotensin antagonists on the rat pressor effect of angiotensin-like substance formed by goosefish Stannius corpuscles and homologous plasma. PMID- 6271621 TI - Insertions of the transposon Tn1 into the Pseudomonas aeruginosa chromosome. AB - The transposon Tn1 has been translocated to the chromosome of Pseudomonas aeruginosa from plasmid R18, following hydroxylamine mutagenesis of the plasmid. Twelve insertions were mapped to six distinct sites distal to 55 min of the origin of chromosome transfer by the plasmid FP2. These map locations were confirmed by host chromosome mobilization tests mediated by plasmids R18 or R91 5, due to Tn1 homology between plasmid and host chromosome. All the Tn1 chromosomal inserts were retransposable to other plasmids (Sa, R931 and R38). The behavior of Tn1 in P. aeruginosa was very similar to its behavior in Escherichia coli with respect to regional specificity, orientation of insertion and in serving as regions of homology for host chromosome mobilization by plasmids. This last property has permitted the demonstration that Tn1 on R18 and R91-5 is in opposite orientation with respect to the origin of transfer (oriT) of the two plasmids. PMID- 6271622 TI - Mating-type differentiation by transposition of controlling elements in Saccharomyces cerevisiae. AB - The nonfunctional mutation of the homothallic gene HML alpha, designated hml alpha, produced two mutant alleles, hml alpha-1 and hml alpha-2. Both mutant clones were mixed cultures consisting of a mating-type cells and nonmating haploid cells. The frequencies of the two cell types were different, and a few diploid cells able to sporulate were found in the hml alpha-2 mutant. Conversions of an a mating-type cell to nonmater, and vice versa, were observed in both mutants. The conversion of an a mating phenotype to nonmating is postulated to occur by alteration of the a mating type to the sterile mating-type allele in the hml alpha-1 mutant. In tetrad dissection of prototrophic diploids that were obtained by rare mating of hml alpha-1 mutants with a heterothallic strain having the MATa ho HMRa HMLa genotype, many mating-deficient haploid segregants were found, while alpha mating-type segregants were observed in a similar diploid using an hml alpha-2 mutant. The mating-type-deficient haploid segregants were supposed to have the sterile alpha mating-type allele because the nonmating genetic trait always segregated with the mating-type locus. Sporogenous diploid cells obtained in the hml alpha-2 mutant clone had the MATa/MAT alpha HO/HO HMRa/HMRa hml alpha-2/hml alpha-2 genotype. These observations suggested that the hml alpha-1 allele produces a transposable element that gives rise to the sterile alpha mating type by transposition into the mating-type locus, and that the hml alpha-2 allele produces an element that provides alpha mating-type information, but is defective in the structure for transposition. PMID- 6271623 TI - [Genetic control of plasmid R6K conjugativity]. AB - The regions determining conjugation ability of plasmid R6K were localized by means of deletion mutants obtained in vitro and in vivo and Tra- mutants induced by integration of transposons Tn5, Tn7 and Tn9 into different DNA sites of the conjugative deletion mutant pAS3. At least 13 genes were found to be involved in the genetic control of R6K conjugativity, on the basis of genetic, restriction and heteroduplex studies. They were mapped within the two DNA regions having the total molecular weight of about 10-11 md. A transposon-like structure with a replication function has been located between them. PMID- 6271624 TI - Recombination between satellite phage P4 and its helper P2. I. In vivo and in vitro construction of P4: :P2 hybrid satellite phage. PMID- 6271625 TI - Recombination between satellite phage P4 and its helper P2. II. In vitro construction of a helper-independent P4: :P2 hybrid phage. PMID- 6271626 TI - Two-dimensional display of lambda and Escherichia coli restriction fragments separated by Hg-Cs2SO4 centrifugation and gel electrophoresis. AB - EcoRI restriction fragments derived from the DNA of bacteriophage lambda and Escherichia coli were fractionated by density gradient centrifugation of their mercury complexes in Cs2SO4 and subsequent electrophoresis on a horizontal agarose-gel slab. In this two-dimensional display, lambda fragments were resolved into six components and E coli fragments into more than 108 components. Bacterial chromosome regions contiguous to lambda prophage integrated at different sites were amplified by induction, and the EcoRI fragments were subjected to the two dimensional analysis. As expected, the sets of amplified fragments were clearly different among the various lysogens. The approximate genome region affected by induction was estimated as one-tenth of the whole chromosome. PMID- 6271627 TI - Molecular cloning of the SUF2 frameshift suppressor gene from Saccharomyces cerevisiae. AB - A genetic approach to the molecular cloning of frameshift suppressor genes from yeast is described. These suppressors act by suppressing +1 G:C base-pair insertion mutations in glycine or proline codons. The cloning regimen involves an indirect screen for yeast transformants which harbor a functional suppressor gene inserted into the autonomously replicating "shuttle" vector YEp13, followed by transfer of the hybrid plasmid from yeast into Escherichia coli. Using this procedure a 10.7-kb DNA fragment carrying the SUF2 frameshift suppressor gene has been isolated. This suppressor acts specifically on +1 G:C insertions in proline codons. When inserted into an integrative vehicle and reintroduced into yeast by transformation, this fragment integrates by homologous recombination in the region of the SUF2 locus on chromosome III. A large proportion of the fragment overlaps with another cloned DNA segment which carries the closely linked CDC10 gene. The SUF2 fragment carries at least two tRNA genes. The SUF2 gene and one of the tRNA genes are located on a 0.85-kb restriction fragment within the 10.7-kb segment. A method is also described for the isolation of DNA fragments carrying alternative alleles of the SUF2 locus. Using this procedure, the wild-type suf2+ allele has been cloned. PMID- 6271629 TI - Cloning of a recA-like gene of Proteus mirabilis. AB - A gene of Proteus mirabilis that can substitute for functions of the recA gene of Escherichia coli has been cloned into the plasmid pBR322, using shotgun experiments. The recA-like gene (recAp.m.) has been localized by restriction mapping within a 1.5-Md PstI fragment that is a part of two cloned HindIII fragments of the chromosome of P. mirabilis. The restriction map of the recAp.m. gene differs from that of the recA gene of E. coli. Functionally, the recombinant plasmids containing the recAp.m. gene restore a nearly wild-type level of UV resistance to several point and deletion mutants in the recA gene of E. coli. PMID- 6271628 TI - The plasmid cloning vector pBR325 contains a 482 base-pair-long inverted duplication. AB - The plasmid pBR325 is a cloning vector constructed in vitro by addition of the chloramphenicol resistance (Cmr) gene of an IS1-flanked transposon to pBR322 (Bolivar, 1978). It is a 5 995 bp plasmid carrying no sequence originating from IS1. DNA-sequence data suggest that its Cmr segment was derived from a Cm transposon longer than Tn9. The plasmid pBR325 carries between the Cmr and Tcr genes a 482 bp sequence which duplicates, in the opposite orientation, a section pf pBR322 located at the end of the tcr gene. The same structure was found in pBR328, a deletion derivative of pBR325 (Soberon et al., 1980). The possible implications of this inverted duplication on cloning experiments are discussed. PMID- 6271630 TI - Restriction maps of plasmids pUB110 and pBD9. PMID- 6271631 TI - Clone banks of the mung bean, pea and spinach chloroplast genomes. AB - All but one of the PstI restriction fragments from mung bean, pea, and spinach chloroplast DNAs have been stably cloned into pBR322. Large fragments (15-54 kb) were cloned at low efficiencies which decreased with increasing fragment length. However, plasmids containing fragments above 25-30 kb were too unstable to be useful. In particular, pBR322 derivatives containing the largest mung bean and spinach fragments (34 kb and 54 kb, respectively) are extremely unstable and rapidly delete parts of the plasmid sequence. The PstI fragments of mung bean chloroplast DNA which cover the 34-kb PstI fragment have been cloned into pACYC177. After a search of several thousand recombinants we were unable to recover a clone containing a 12.2-kb pea chloroplast PstI fragment and suggest that some property of its sequence may be inimical to the cloning process. The identity of the cloned fragments to native chloroplast DNA restriction fragments is demonstrated by restriction analysis and the ability to construct detailed restriction maps of the mung bean and pea chloroplast genomes. PMID- 6271632 TI - One-step gene amplification by Mu-mediated transposition of E. coli genes to a multicopy plasmid. PMID- 6271633 TI - Plasmid vectors for high-efficiency expression controlled by the PL promoter of coliphage lambda. PMID- 6271634 TI - A plasmid cloning vector for the direct selection of strains carrying recombinant plasmids. PMID- 6271635 TI - Plasmid cointegrates and their resolution mediated by transposon Tn3 mutants. PMID- 6271636 TI - The nucleotide sequence surrounding the promoter region of colicin E1 gene. AB - The nucleotide sequence of 570 bp, covering the N-terminal portion of the colicin E1 gene, was determined. The sequence of the N-terminal four amino acids of the colicin E1 protein, determined by manual Edman degradation, agreed with that predicted from the nucleotide sequence. From analysis of the 5'-terminal sequences of RNAs synthesized in vitro, the promoter and operator regions of the colicin E1 gene were assigned. These data indicate the existence of two promoters, one of which is located in the coding region for colicin E1. DNA sequence homology of 16 bp was found between the putative operator regions of the colicin E1 and recA genes. PMID- 6271637 TI - Expression of a cloned Saccharomyces cerevisiae gene (URA1) is controlled by a bacterial promoter in E. coli and by a yeast promoter in S. cerevisiae. AB - The expression of a cloned yeast URA1 gene in Escherichia coli and in Saccharomyces cerevisiae was studied. In E. coli, only one orientation of the cloned yeast DNA segment inserted into the bacterial vector (pBR322) allows URA1 expression. Moreover, the permissive orientation changes with the cloning site. The absence of URA1 expression in E. coli can be corrected by the spontaneous integration into the cloned yeast DNA of a 0.9-kb bacterial DNA. Several copies of such a bacterial IS element have been detected in the host E. coli genome. The results strongly suggest that, in E. coli, transcription of the yeast URA1 needs a prokaryotic promoter for its initiation. In S. cerevisiae, the expression of non-chromosomally cloned URA1 does not depend on the orientation of the cloned fragment. Furthermore, it remains under the control of a nuclear regulatory gene (pprX-1) which constitutively enhances the expression of URA1 as well as URA3 at the transcriptional level. Therefore, in S. cerevisiae, transcription of non chromosomally cloned URA1 involves a physiological yeast promoter cloned along with the structural part of the gene. PMID- 6271638 TI - Transposition of a tetracycline-resistance determinant (Tn1523) and cointegration events mediated by the pIP231 plasmid in Escherichia coli. AB - A 10.8-kb transposable DNA sequence conferring resistance to tetracycline resides on the IncY Escherichia coli plasmid pIP231. This sequence, designated Tn1523, was shown to insert into different sites of the replicons of the IncY prophage P1Cm c1.100 and the IncI1 plasmid pIP112. This process is not dependent on the host recombination system recA. Genetic results indicate that Tn1523 transposition involves the formation of a cointegrate intermediate, either between pIP231 and P1Cm c1.100, or between pIP231 and pIP112. These intermediates were found to be resolved into donor and recipient plasmids, each harboring a copy of the Tn1523 transposon. A stable structure formed by fusion of the pIP231 plasmid with the pIP112 plasmid was also observed. This event occurs in the absence of the bacterial recA gene product and seems to involve a site-specific reciprocal recombination between "IS-like" elements. PMID- 6271640 TI - Modification of the bacteriophage vector M13mp2: introduction of new restriction sites for cloning. AB - The construction of two new derivatives of the bacteriophage cloning vector M13mp2 is described. One derivative, mWJ22, contains a new HindIII site while the other, mWJ43, contains a new BamHI site. These new sites were both introduced at the EcoRI site at amino acid five of the 145 amino acid-long fragment of Escherichia coli beta-galactosidase within the phage. The new restriction sites do not disrupt the blue color detection system of M13mp2; therefore insertion of cloned fragments results in colorless plaques on indicator plates for the new derivatives. PMID- 6271639 TI - DNA sequence of the att region of coliphage 434. AB - Phages lambda and 434 are related phages that insert at the same site on the Escherichia coli chromosome. A 5.9-kb SalI-BamHI fragment derived from phage 434 was shown to hybridize to a 0.5-kb probe carrying attP-lambda. A 0.8-kb Bam HI TaqI fragment subcloned into pBR327 was used for sequencing. The sequence of the 500 bp around the insertion site is given here, Comparison of the lambda and 434 sequence shows that the following regions are conserved: the coding sequence for the integrase protein (only 162 bp have been sequenced corresponding to the carboxy terminus), the 15-bp common core at the insertion site, and the three integrase-binding sites flanking the insertion site. The lambda and 434 sequences diverge radically to the left of base-197, suggesting that DNA to the left of that point plays no specific role in insertion or its regulation. PMID- 6271641 TI - Human ribosomal RNA gene spacer sequences are found interspersed elsewhere in the genome. AB - A cloned EcoRI fragment containing human 18 S rRNA gene sequences was used to screen a gene library to obtain a set of 8 overlapping cloned DNA segments extending into the non-transcribed spacer region of the human ribosomal RNA gene cluster. 19.4 kb of the approx. 43-kb rDNA repeat was obtained in cloned form and mapped with restriction endonucleases. None of the clones obtained extended into 28 S rRNA sequences. A 7-kb region of non-transcribed spacer DNA shared in common between five independent clones was subjected to comparative restriction digests. It was estimated that sequences among the five different spacer isolated varied by not more than 1.0%, if all the observed differences are assumed due to point mutation. HaeII-restriction fragments from within this same 7-kb region contain sequences carried not only within the tandem repeats of the gene cluster but interspersed elsewhere in the genome. Some of these sequences correspond to the Alu family of highly repeated interspersed sequences. PMID- 6271642 TI - Variants of a cloned synthetic lactose operator. II. Chloramphenicol-resistant revertants retaining a lactose operator in the CAT gene of plasmid pBR325. PMID- 6271643 TI - The identification and partial characterization of a plasmid containing the gene for the membrane-associated hydrogenase from E. coli. AB - Escherichia coli DNA was digested with restriction endonuclease PstI and ligated into the PstI site of plasmid pBR322. Recombinant plasmids that were constructed in this manner were used to transform E. coli H61, a mutant with a decreased level of hydrogenase activity. Complementation of this hydrogenase mutation identified a bacterial clone carrying the gene for the membrane-associated E. coli hydrogenase in plasmid pBL101. In E. coli minicells, the pBL101 DNA directed the synthesis of a protein of a size corresponding to that of the precursor of the E. coli membrane-associated hydrogenase, which appears to contain an uncleaved leader peptide. A restriction map of the cloned DNA was determined for 14 endonucleases. PMID- 6271645 TI - Dispensable sequences and packaging constraints of DNA from the Streptomyces temperate phage phi C31. AB - Deletion mutants of the temperate Streptomyces phage phi C31 were selected by two methods: resistance to the chelating agent sodium pyrophosphate, and plating of a phi C31::pBR322 hybrid phage on Streptomyces albus G to obtain large plaque mutants. The deletions defined a 7.7 kilobase (kb) segment of the phi C31 genome which is inessential for plaque formation, in addition to a shorter segment including the repressor gene. Analysis of deletions and insertions suggested that the minimum size of the phi C31 genome allowing plaque formation is 37.5 kb (91% of the wild-type length of 41.2 kb), and the maximum is at least 42.4 kb (103%). These results indicate that it should be possible to introduce up to 10 kb of foreign DNA into a suitably developed phi C31 vector. PMID- 6271644 TI - Characterization of the nuclear ribosomal DNA of Euglena gracilis. AB - A phage lambda recombinant library containing Euglena gracilis genomic DNA was screened for nuclear rDNA sequences. A recombinant phage was isolated that contained an 11.5-kb nuclear rDNA sequence. The 11.5-kb insert was mapped with restriction endonucleases and was shown to represent a complete rDNA repeat unit that carried the genes for the 19S, 25S, 5.8 S and 5 S cytoplasmic rRNAs. The 2000 rDNA repeat units per haploid genome are organized in the form of identical tandem repeats. PMID- 6271646 TI - Cloned fragments of human adenovirus type-12 DNA. AB - The following restriction endonuclease fragments of human adenovirus type 12 (Ad12) DNA have been cloned in plasmid or bacteriophage lambda vectors using standard protocols: the EcoRI-A*, -B, -D, -E, and -F fragments, the BamHI-B, -C, D, -F, -G, -H, and -I fragments, the HindIII-F and -I fragments, and the PstI-A, D, -F, -G, and -H fragments. The EcoRI-A* fragment comprises the right terminal 5 kb of Ad12 DNA including the terminal 143 bp. PMID- 6271647 TI - [Standards for chemical substances depending on the type of soil]. PMID- 6271648 TI - [Selective sampling of carbon monoxide on a zeolite adsorbent]. PMID- 6271649 TI - [Apropos the article by N. Ia. Ianyshevaia, I. A. Chernichenko, N. V. Balenko and N. A. Pavlova, " The Effect of Exposure Duration and size of a Single Benzo(a) pyrene Dosage on the Nature and Intensity of the Carcinogenic Reaction"]. PMID- 6271650 TI - [Parabiosis: the basis for general biological reactions to the harmful action of toxic substances]. PMID- 6271651 TI - Pressure changes after balloon distension of the colon wall in diverticular disease. AB - Balloon distension of the distal colon in diverticular disease does not produce the pressure change in response to increasing volumes that occurs in normal subjects. This phenomenon, though modified at first by resection, is not abolished by resection nor by myotomy or bran. This adaptive quality of the wall of the colon to balloon distension possibly reflects a structural change in its wall, as the phenomenon was present in the colon in diverticular disease in vitro as well as in vivo. PMID- 6271652 TI - Cytomegalovirus inclusions in the gastroduodenal mucosa of patients after renal transplantation. AB - Biopsies collected from gastroduodenal mucosa during endoscopic examination of 20 patients having undergone renal transplantation and subsequent immunosuppressive therapy showed cytomegalovirus (CMV) inclusion bodies in nine cases. CMV antibody titres were tested in all patients before and after the transplant procedure. Not all patients exhibited viraemia-related symptoms at the time of endoscopy. No correlation was found between the presence of CMV-type cells within the gastroduodenal mucosa, endoscopic and histological findings, the duration of the transplant, and the dosage of immunosuppressive drugs. The duodenum seems to be the elective site of CMV. The involvement of gastric mucosa seems to represents a worsening of the illness. Eight of nine patients with positive biopsies for CMV inclusion had negative pretransplant antibody titres to CMV. All nine patients were seropositive after transplantation and showed seroconversion. Five of 11 recipients with negative biopsies for CMV inclusion bodies, were seronegative before transplantation. Seroconversion occurred in five patients after the transplant; the other six had no rise in antibody titres. The lack of pre transplant CMV antibody titre and its subsequent increase after transplantation identifies a greater risk of developing post-transplant CMV infection. PMID- 6271653 TI - Recurrent gestational trophoblastic disease. PMID- 6271655 TI - [Hormone receptors: mode of action and clinical importance]. PMID- 6271654 TI - Trophoblastic pseudotumor-evidence of malignant disease potential. PMID- 6271656 TI - [Growth inhibiting therapy in osteochondrosis tibiae - Blount disease]. AB - Therapy with estrogens was inaugurated in a girl with osteochondrosis tibiae in order to accelerate the fusion of epiphyseal lines. This therapy can be regarded as additional therapy but orthopedic therapy is necessary once growth of the patient has stopped. PMID- 6271657 TI - [Myelography. Contrast media, indications, complications]. PMID- 6271658 TI - [Urologic emergencies in children]. PMID- 6271659 TI - Effects of thyroid hormone and adrenalectomy on [Na+ + K+]ATPase in the ob/ob mouse. AB - The absence of the thyroid stimulation of hepatic [Na+ + K+]ATPase in obese (ob/ob) mice has been investigated. A wide range of tri-iodothyronine (T3) concentrations failed to enhance the hepatic [Na+ + K+]ATPase of ob/ob mice made hypothyroid by methimazole treatment but glycerophosphate dehydrogenase activity was maximally stimulated at low doses of T3. Adrenalectomy increased the basal activity of [Na+ + K+]ATPase to levels found in lean control mice and restored the response of this enzyme to T3. Body weight gain was unaffected by the induction of hypothyroidism in either lean or ob/ob mice. It is concluded that the adrenal steroids play an important role in the expression of [Na+ + K+]ATPase activity in the ob/ob mouse. PMID- 6271660 TI - Further characterization of human basic-somatomedin: comparison with insulin-like growth factors I and II. AB - Our basic-somatomedin (SM) was further compared with insulin-like growth factors (IGF) I and II. Basic-SM and IGF revealed similar sulfation factor (SF) activity in cartilage, insulin-like activity(ILA) in adipocytes, and receptor binding activity to adipocytes and placental cell membranes.IGF-II revealed less SF activity but more ILA than basic-SM. Comparison of SM and insulin in terms of ILA and binding activity to adipocytes suggested that adipocytes have separate insulin and SM receptors and that the ILA of SM is mediated through the SM receptors. These studies also suggest that the receptors for acidic-neutral group of SM mediate the ILA of SM, whereas the growth promoting effects of SM are mediated via receptors for the basic group of SM. PMID- 6271661 TI - Degradation during storage of high molecular weight immunoreactive insulin (HWIRI) and proinsulin (PLC) in extracts of Benign insulinomas and adjacent pancreatic tissue. PMID- 6271662 TI - Gonadotropic hormone binding to human ovarian tumors. AB - A variety of normal human ovarian tissues as well as benign and malignant human ovarian neoplasms have been examined in vitro for the presence of gonadotropin binding sites and gonadotropin-stimulable cAMP production, in order to determine whether gonadotropic hormones have an effect on ovarian tumors. Binding of either FSH or hCG, or both, was demonstrated in several tumors from each of the histologic subgroups of primary ovarian tumors, including epithelial, sex cord stromal, and germ cell types. Cyclic AMP stimulation was found in one tumor of sex cord-stromal origin, in two of epithelial origin, in two of germ cell origin, and in one tumor-like condition, pregnancy luteoma. Additional biochemical studies are needed to characterize receptor sites and to relate them to histologic features, growth behavior in vivo, and serum gonadotropin levels. PMID- 6271663 TI - Ultrastructural study of osteosarcomas. AB - The ultrastructural examination of four osteosarcomas (osteogenic, undifferentiated, and pleomorphic) is described. There are three types of tumor cells. Most of the cells are held in contact by desmosome-like tight junctions; they are atypical osteoblasts with cytoplasmic processes, dilated rough endoplasmic reticulum, mitochondria carrying calcific inclusions, lipid droplets surrounded by glycogen, and intracellular fine filamentous fibers. Other cells exhibiting varying degrees of osteoblastic maturity are also seen with multilobed nuclei, a clear cytoplasm, and straight bordered membranes. The last type is chondroid with abundant deposits of glycogen, lipid droplets, and undilated rough endoplasmic reticulum. The matrix is composed of fibrils without periodicity, scattered and deteriorated collagen fibers, and focal calcium deposits of hydroxyapatite crystals as in embryonal bone, dentine, or callus bone. PMID- 6271664 TI - An unusual pelvic tumor with benign glandular, sarcomatous, and Wilms' tumor-like components. AB - We present a pelvic tumor occurring in a 23 year old woman in which a Wilms' tumor-like element was predominant. The presence of a benign glandular component surrounded by small spindle and round malignant cells paralleled that encountered in adenosarcoma of the uterine body and cervix. The possible histogenesis is discussed. Clinically the recurrent tumor showed evidence of a response to chemotherapy. Treatment was complicated by petit mal seizures, for which adriamycin was responsible. The patient exhibited no evidence of disease 22 months after the first operation. PMID- 6271665 TI - Possible role of genetic cell variants in the viral induction of mouse mammary tumors. AB - Out of three attempts to induce neoplasia in normal C57Bl mammary epithelial cells with the mouse mammary tumor virus (MuMTV) only one presented signs of tumorigenicity. Immunofluorescence showed that virus synthesis took place in all three sublines but tumorigenicity as detected by cell aggregation viability (CAV) and transplantation into syngeneic mice failed to occur in two of them. By comparison, cells from a BALB/c spontaneous mammary tumor that do not express MuMTV were 100% tumorigenic, whereas cells from a BALB/cfC3H tumor with a 95% virus-producing cell population had a normal CAV and were tumorigenic only in 60% of the test animals. This lack of correlation suggested that many of the virus producing cell were not neoplastic and that neoplasia might occur under virus stimulation only if a restricted population of genetic cell variants existed. Accelerated tissue culture passages of virus-free C57Bl and BALB/c normal mammary cells resulted in their spontaneous neoplasia at Passages 23 and 50, respectively; when duplicated cells cryopreserved in early passages were revived and cultivated in the same manner, neoplasia occurred at Passages 27 and 58. The similarity of the passage numbers appears to confirm the existence of genetic cell variants among the normal cell population. PMID- 6271666 TI - Characterization of the bone marrow cell population expressing an AKR murine leukaemia-virus gp71-like antigen. AB - The murine bone marrow population mainly responsible for the unique competing activity of this lymphoid organ in the competition radioimmunoassay (cRIA) for AKR murine leukaemia viruses gp71 has been partially purified and characterized. That population could be distinguished by the following properties: (i)gp71+ cells (cell population expressing the AKR MuLV gp71 antigen) are mainly medium density cells, according to their distribution in the layers of a discontinuous bovine serum albumin (BSA) gradient; (ii) The gp71 positive cells are not mature lymphocytes; (iii) This cell population is found both in the FcR+ and FcR- compartments and is not included in the complement (C') receptor-positive population; and finally, (iv) the gp71+ cells are phagocytic cells and an adherent cell population according to its capacity to adhere to plastic surfaces, nylon wool and Sephadex G-10. PMID- 6271667 TI - Specific changes in hypothalamic alpha-adrenoceptors in young spontaneously hypertensive rats. AB - Changes in the activity of hypothalamic and brain-stem adrenergic neurons have been reported in young spontaneously hypertensive rats (SHR) prior to the development of hypertension. We have measured central alpha- and beta adrenoceptor concentrations in 4-week-old SHR and Wistar-Kyoto (WKY) controls by direct radioligand binding studies using [3H]prazosin (alpha 1), [3H]clonidine (alpha 2), and [125I]iodohydroxybenzlpindolol (beta). The concentration of alpha 2-adrenoceptors was significantly elevated in the hypothalamus of the SHR, 156.9 +/- 10.4 compared with WKY, 119.4 +/- 10.0 fmole/mg protein (n = 7, mean +/- SEM, p less than 0.0125). Alpha 2-adrenoceptor concentrations in both the brain stem and cerebral cortex were similar in the two groups of animals. The increase in hypothalamic adrenoceptors was specific for alpha 2-adrenoceptors, since similar concentrations of alpha 2- and beta-adrenoceptors were found in this region. PMID- 6271668 TI - Acute and chronic intrarenal alpha- and beta- adrenergic receptor stimulation of renin release in the conscious dog. AB - The effect of continuous intrarenal infusion of norepinephrine, isoproterenol, and methoxamine on renin release was studied in the uninephrectomized conscious dog. Chronic intrarenal infusion of norepinephrine produced a biphasic curve of plasma renin activity (PRA) and a sustained 25 mm Hg increase in mean arterial pressure (MAP). The initial increase in PRA peaked at 3 hours, after which PRA returned to control levels. Alpha- or beta-adrenergic antagonists did not attenuate the initial rise in PRA. The PRA increased again after 48 hours of chronic intrarenal norepinephrine infusion and remained elevated thereafter. The second rise in PRA was increased by 30% with alpha-adrenergic blockade. Chronic intrarenal isoproterenol administration produced a similar increase in PRA, which peaked at 3-5 hours and then returned to control levels. In contrast to norepinephrine, chronic isoproterenol administration did not result in a second increase in PRA. At the end of the chronic isoproterenol infusion period, beta adrenergic receptor refractoriness was demonstrated, as PRA did not increase significantly in response to a fourfold increase in the dose of isoproterenol. An increase in PRA was produced by acute intrarenal infusion of methoxamine. This increase in PRA was blocked by phentolamine, suggesting a vascular alpha adrenergic receptor-mediated release of renin. PMID- 6271669 TI - Cation fluxes and Na+-K+-activated ATPase activity in erythrocytes of patients with essential hypertension. AB - Recently it has been claimed that the active potassium influx in erythrocytes of patients with essential hypertension would be increased. In view of the diagnostic and possibly therapeutic potential of this claim, we have determined the Na+-K+ activated ATPase activity and the affinity of the enzyme for Na+, K+, and ATP in membranes isolated from erythrocytes of hypertensive (with and without medication) and normotensive subjects. Subsequently, the active (ouabain sensitive) sodium and potassium fluxes and their ratios have been determined after treatment of intact erythrocytes either with cold or with p chloromercuribenzene-sulfonate (PCMBS). Finally, in view of a subsequent claim that the furosemide-sensitive, ouabain-insensitive cation fluxes would be greatly reduced in erythrocytes of patients with essential hypertension, we have determined these fluxes in choline chloride medium containing ouabain with and without furosemide. For none of these parameters has any significant difference between hypertensive and normotensive subjects been found except for a decrease in the ouabain-sensitive K+ influx after cold treatment in hypertensives. This is also true for the hypertensive subjects who had a known hypertensive parent. It is concluded that the results do not support a role of Na+-K+ activated ATPase or the furosemide-sensitive cation carrier in the pathogenesis of essential hypertension, and that ouabain-sensitive and furosemide-sensitive cation flux determinations in erythrocytes do not seem to be useful for the diagnosis of this condition. PMID- 6271670 TI - Left ventricular hypertrophy in rats with renovascular hypertension. Alterations in cardiac function and adrenergic responses. AB - Performance of the hypertrophied left ventricle was studied by determination of the inotropic response to different stimuli in renal hypertensive rats (two kidney, one clip Goldblatt, RHR, n = 13) and matched sham-operated controls (NR, n = 11). A model was developed to determine maximal pressure development (transient aortic ligation), maximal pumping ability (rapid transfusion, 2 ml/30 sec), and responses to beta stimulation (isoproterenol, 0.01 to 0.10 micron g/kg/min), using dP/dt/P40 as a load-independent index of contractility. With rapid blood transfusion, RHR developed a higher ventricular systolic pressure (211.5 +/- 10.1 mm Hg vs 194.0 +/- 9.3 (SE), p less than 0.001) but at the expense of higher end-diastolic pressure (LVEDP) (12.2 +/- 1.1 mm Hg vs 7.7 +/- 1.0, p less than 0.02). The maximal response of dP/dt/P40 to isoproterenol was diminished in RHR (29.5 +/- 3.2 sec-1 vs 49.6 +/- 5.2, p less than 0.01) whereas the maximal developed pressure (MDP) was greater in RHR than in NR (239.2 +/- 7.5 mm Hg vs 197.0 +/- 3.9, p les than 0.001). A positive correlation was found between MDP and ventricular weight (r = 0.846, p less than 0.001) in contrast with the negative correlation found between ventricular weight and maximal dP/dt/P40 response to isoproterenol (r = 0.677, p less than 0.001). Thus, cardiac hypertrophy in RHR allowed higher developed ventricular pressures but at the expense of higher LVEDP; at the same time, however, the ability to increase contractility in response to beta adrenergic stimulation was decreased. The contrast in results obtained using different tests of cardiac function indicates the need for a multifactorial approach. It also suggests a subtle transformation in this hypertrophy of the pattern of cardiac adaptation to the increased load. PMID- 6271671 TI - Long-term converting enzyme inhibition and sympathetic nerve function in hypertensive humans. AB - Orthostatic hypotension is uncommon during oral converting enzyme inhibition, even when combined with salt depletion. To assess the mechanisms responsible for the cardiovascular homeostasis in this condition, we studied the blood pressure (BP), heart rate (HR), total plasma catecholamines (CA), and plasma renin activity (PRA) responses after 20 minutes of 60 degrees head-up tilt in four groups of hypertensive patients. Group 1 included seven untreated patients; Group 2, eight patients on converting enzyme inhibitor (captopril) therapy; Group 3, six patients on diuretic therapy and Group 4, 15 patients on combined captopril and diuretic therapy. Long-term converting enzyme inhibition alone or in combination with diuretics resulted in reduction of mean arterial pressure (MAP) associated with a marked increase in PRA and fall in plasma aldosterone concentration (PAC). Pronounced increases in HR and plasma CA on tilt were observed in all groups. In Groups 1, 2, and 3, BP was maintained during tilt; in Group 4, three patients fainted between 5 and 15 minutes while the other 12 had a normal response to tilt. Plasma catecholamines increased more significantly after 15 and 20 minutes of tilt, more in Groups 3 and 4 than in Group 1, while no differences in HR response were observed among groups. Results suggest that sympathetic compensatory mechanisms are adequate in the majority of patients to maintain BP during converting enzyme inhibition even when combined with salt depletion. In a few who exhibited orthostatic hypotension, a vasovagal attack seemed to be responsible for bradycardia and fall in BP. PMID- 6271672 TI - Active and inactive kallikrein released by kidney slices from normotensive and hypertensive rats. AB - The relative rates of release of active and inactive kallikrein were investigated in the rat kidney by studying in vitro kidney slices. The slices were kept in Warburg flasks in a Krebs-Ringer solution for 15 up to 60 minutes, and their kallikrein content and the kallikrein-like activity released by them were measured at different times. Renal kallikrein concentration from two-kidney, one clip hypertensive rats was less than in the sham-operated controls (p less than 0.01). Active kallikrein released into the bathing medium was 4.7 +/- 0.3 ng/mg/hr in normotensive rats (n = 10) and 3.3 +/- 0.3 in two-kidney hypertensive animals (n = 12); the difference between both groups was statistically significant (p less than 0.01). Inactive kallikrein is between 60% and 70% of the total kallikrein released by kidneys of normal and hypertensive rats. Thus, under the conditions of these experiments, the kidney slices were able to release a kallikrein-like enzyme in an active and an inactive form. The rate of active kallikrein released from slices of hypertensive animals was lower than in the normotensive controls. PMID- 6271673 TI - Kallikrein and prekallikrein on the basolateral membrane of rat kidney tubules. AB - Basolateral membrane (BLM) enriched fraction was isolated from homogenized rat kidney cortex by differential centrifugation. We also obtained a fraction enriched in plasma membrane (PM). The morphology of the isolated BLM fragments was studied by transmission and freeze fracture electron microscopy. The relative specific activity of Na+-K+-ATPase was enriched 7-fold, while that of marker enzymes for PM, endoplasmic reticulum, and lysosomes was lower than in the crude homogenate. There was a 10-fold difference in the ratios of activities of Na+-k+ ATPase to Mg2+-ATPase in the BLM and in the PM enriched fractions. Kallikrein activity was determined with S-2266 substrate and by radioimmunoassay of kinin released. It was low in the BLM fraction prior to adding detergent, but Triton X 100 increased the activity 12 to 16-fold. Both free trypsin and Sepharose 4B bound insoluble trypsin increased kallikrein activity 2- to 3-fold in both the membrane-bound and soluble fractions, probably by activating a prekallikrein. The results were interpreted that the kallikrein studied originated from the distal tubular BLM. PMID- 6271674 TI - Effects of tonin on the adrenal secretion in the rat. AB - Tonin infused intravenously at a rate of 12 micron g/kg/min for 2 hours stimulated aldosterone and corticosterone secretion in conscious rats. In pentobarbital-anesthetized rats treated with dexamethasone and morphine, tonin stimulated aldosterone secretion, but corticosterone was unaffected. Tonin did not stimulate steroidogenesis in isolated rat adrenal glomerulosa cells unless substrate, in the form of "stripped" rat plasma, was added, in which case large amounts of immunoreactive angiotensin II were generated. These results suggest that tonin may generate an angiotensin peptide in vivo, which in turn stimulates aldosterone secretion. ACTH is secreted in response to tonin infusion, and contributes to the adrenal response. PMID- 6271675 TI - Properties of calcium ionophore-induced generation of superoxide anion by human neutrophils. AB - Exposure of human neutrophils to the calcium ionophore, A23187, eventuates in a time- and concentration-dependent generation of superoxide anion (O2-) in the presence but not absence of extracellular calcium. The selective requirement for calcium is demonstrated by the observation that magnesium caused a dose-related inhibition of A23187-stimulated O2- generation. Preincubation of neutrophils with cytochalasin B prior to their interaction with A23187 results in a significant enhancement of O2- production. The activity of the O2--generating system was maximum at 37 degrees C and was significantly curtailed at lower and higher temperatures. A23187-induced O2- generation was inhibited by the sulfhydryl reagents. N-ethyl maleimide (NEM) and iodoacetic acid (IA), and by the metabolic inhibitor 2-deoxy-D-glucose (2-DG) in the absence of glucose and cytochalasin B. Cyanide was inactive. Therefore, A23187 represents a useful pharmacologic probe for investigating the divalent cation and metabolic requirements of the neutrophil O2--generating system. PMID- 6271676 TI - Functional and metabolic properties of alveolar macrophages in response to the gas phase of tobacco smoke. AB - The effect of whole tobacco smoke and the gas phase of tobacco smoke on the metabolism and phagocytic ability of alveolar macrophages was monitored over a 30 day exposure period. It was demonstrated that both the gas phase and whole tobacco smoke induced a weight loss in exposed rats. Alveolar macrophage oxygen consumption was markedly increased by both exposure regimens. Superoxide generation was not affected by whole tobacco smoke exposure but was increased in response to the filtered gas phase. Hexose monophosphate shunt activity was not altered by either treatment. When metabolic alterations were seen in response to the separate exposures, they were seen only after a phagocytic challenge to the macrophage and not when the cell was unchallenged. Neither whole tobacco smoke nor the gas phase had any significant effect on the ability of alveolar macrophages to phagocytize a viable challenge of Staphylococcus aureus. Our results suggest that many of the metabolic and functional effects of tobacco smoke on alveolar macrophages can be attributed to the gas-phase component of whole tobacco smoke. PMID- 6271677 TI - Mitogen- and viral antigen-induced transformation of lymphocytes from normal mink and from mink with progressive or nonprogressive Aleutian disease. AB - Peripheral blood lymphocytes (PBL) from mink with progressive Aleutian disease (AD) were shown to be significantly less responsive to phytohemagglutinin, concanavalin A, and pokeweed mitogen than were PBL from normal mink and from mink with a nonprogressive form of AD. Response to the virus of AD was significantly greater in PBL cultures from mink with nonprogressive AD than in those from normal mink or mink with progressive AD. After experimental infection with AD virus, mink PBL were responsive to viral antigen only transiently. These findings suggest that lymphocyte responsiveness as indicated by transformation induced by mitogens or viral antigen may be an important aspect of host response to infection with the parvovirus of AD. PMID- 6271678 TI - Production of exoenzyme S by clinical isolates of Pseudomonas aeruginosa. AB - Exoenzyme S differs from toxin A and diphtheria toxin in that it does not adenosine diphosphate (ADP)-ribosylate elongation factor-2, but rather catalyzes the transfer of the ADP-ribose moiety of nicotinamide adenine dinucleotide to a number of different proteins in extracts of eucaryotic cells. Polyoma-transformed BHK-21 cells were isolated which were resistant to diphtheria toxin and toxin A. Extracts from these cells are ADP-ribosylated by exoenzyme S but not toxin A or diphtheria toxin, providing an assay which distinguishes between S and A activities. A total of 124 clinical isolates of P. aeruginosa were analyzed for production of toxin A and exoenzyme S. Exoenzyme S production was detected in 38% of the strains, whereas 80% of the strains produced toxin A. PMID- 6271679 TI - Specific lymphocyte blastogenic responses in children with cytomegalovirus and herpes simplex virus infections acquired early in infancy. AB - Cell-mediated immune responses in 27 infants and children with cytomegalovirus (CMV) infection acquired between birth and 1 year of age were compared with responses in 13 children who had neonatal herpes simplex virus (HSV) infection. Infection was asymptomatic in 25 of 27 CMV-infected children; the 13 patients with HSV infection were all ill as newborns. The median age when studied was 46 months for children infected with CMV and 24 months for those infected with HSV. We measured lymphocyte transformation responses (LTRs) to CMV antigens in the former group and to HSV type 1 (HSV-1) (and in six cases to HSV-2) in the latter group, with the results expressed as a stimulation index. Based on the results in seropositive and seronegative adult control subjects, stimulation indexes of greater than or equal to 3 were considered indicative of a positive LTR. Among the CMV-infected children, a positive LTR was observed in 0 to 13 assays performed before 1 year of age, 3 of 8 assays performed between 1 and 4 years of age, and 9 of 15 assays performed over 4 years of age. In contrast, a positive LTR to HSV-1 was seen in 15 to 18 assays performed in children under 1 year of age and in 14 of 16 assays performed in survivors of neonatal HSV infection older than 1 year. Six HSV-2-infected patients were tested simultaneously 13 times with HSV-1 and HSV-2 antigens. Those patients under 6 months of age responded similarly to each antigen, whereas those who were older had significantly higher LTRs to HSV-2. Children with CMV infection that was acquired early had persistently diminished specific LTRs. In contrast, after neonatal HSV infection, LTRs to HSV were present even in infancy and became more specific for the infecting type with increasing age. PMID- 6271680 TI - Heterogeneity of rabbit endogenous pyrogens is not attributable to glycosylated variants of a single polypeptide chain. AB - Rabbit endogenous pyrogens were of about the same molecular size, but showed considerable heterogeneity of their isoelectric points. We attempted to show that this heterogeneity was attributable to variable glycosylation of a single polypeptide chain. When peritoneal exudate cells were stimulated to make pyrogens in the presence of 2-deoxy-D-glucose, there was a relatively trivial suppression of pyrogen release, and analysis by isoelectric focusing showed parallel inhibition of secretion of all the forms of endogenous pyrogen. When cells were stimulated in the presence of 3H-labeled amino acids and 14C-labeled glucosamine or glucose, the purified pyrogens were labeled with 3H but not with 14C. Macrophage membrane preparations were made which contained glycosyl transferases and could transfer sugar residues from sugar nucleotides to deglycosylated fetuin. These macrophage membrane preparations did not transfer sugars to the pI 7.3 endogenous pyrogen. Treatment of endogenous pyrogens with neuraminidase or with periodate produced no evidence suggesting that the pyrogens were glycosylated. Last, endogenous pyrogens did not bind to any of four lectins with different carbohydrate specificities. This evidence suggests that the heterogeneity of rabbit endogenous pyrogens is not attributable to glycosylation and must have some other cause. PMID- 6271681 TI - Use of monoclonal antibody directed against herpes simplex virus glycoproteins to protect mice against acute virus-induced neurological disease. AB - Monoclonal antibodies HCl and HD1, directed against herpes simplex virus type 1 (HSV-1) glycoproteins gC and gD, respectively, were evaluated for their ability to passively immunize mice against acute virus-induced neurological disease after footpad inoculation with HSV-1 or herpes simplex virus type 2 (HSV-2). Control virus-infected mice receiving a single intraperitoneal injection of normal serum died within 7 to 10 days after the spread of virus from footpad to spinal cord and brain. However, a single intraperitoneal injection of either HCl or HD1 antibody protected mice from neurological illness and death when administered to HSV-1 (strain HTZ)-infected mice at either 2 h before virus challenge or at 24 h after virus inoculation. To determine the in vivo specificity of the antibodies, passive transfer studies were performed with mice infected with the MP strain of HSV-1, a mutant of HSV-1 (mP) which is defective in the production of glycoprotein gC. Whereas HD1 antibody decreased the incidence of neurological illness in MP- and mP-infected mice, HCl antibody, which protected mP-infected animals, failed to protect mice infected with the MP strain. When HD1 antibody was administered to HSV-2 (strain G)-infected mice at either 2 h before virus challenge or at 6 h (but not 24 h) after virus inoculation, 100% of the infected animals receiving HD1 antibody survived. In contrast, 100% of HSV-2 (strain G) infected animals passively immunized with HCl antibody developed neurological illness and died. These results provide in vivo evidence that the HSV-induced glycoprotein gC expresses type-specific antigenic determinants, whereas glycoprotein gD expresses type-common determinants. PMID- 6271682 TI - Congenital infection of mice with Japanese encephalitis virus. AB - Transplacental transmission of Japanese encephalitis virus (JEV) when given intraperitoneally was demonstrated in pregnant mice as shown by isolation of the virus from placenta and fetal tissues. Furthermore, JEV could be isolated from the brain, liver, and spleen of newborn mice. The effect of JEV at different periods of gestation in pregnant mice was demonstrated for the first time, and the consequences of maternal infection on fetuses and neonates were studied. JEV infection during the 1st week of gestation caused a significantly higher number of fetal and neonatal deaths (66%) than during the 3rd week of gestation (13.8%). The number of abortions, stillbirths, and neonatal deaths was higher in infected mothers than in controls. No congenital abnormalities were found in any of the newborn mice. Sera obtained from 5-week-old health mice delivered by mothers infected during the 3rd week of gestation contained JEV hemagglutination inhibiting and immunoglobulin M antibodies. The results of these preliminary experiments show the usefulness of mice as a model for further elucidation of JEV infection during pregnancy and its effects on the fetus. PMID- 6271683 TI - Parainfluenza 3 virus: plaque-type variants lacking neuraminidase activity. AB - Virus clones lacking detectable neuraminidase activity (SC-YN and M-YN) as well as those possessing it (LT-910N and LT-YN) were isolated from bovine strains of parainfluenza 3 virus. LT-910N and LT-YN viruses produced large turbid plaques in MDBK cells, and SC-YN virus produced small clear plaques. Incorporation of a bacterial neuraminidase in agar overlay medium made SC-YN virus form large turbid plaques, whereas it made M-YN virus form large clear plaques. However, M-YN virus formed only pinhole plaques or no plaques in the absence of neuraminidase. The exogenous neuraminidase had little effect on the plaque formation of LT-910N and LT-YN viruses. M-YN virus induced extensive syncytial formation, and SC-YN virus produced less extensive syncytial formation. The exogenous neuraminidase enhanced replication of SC-YN and M-YN viruses and reduced syncytial formation by these viruses. The enzyme had little effect on replication and cytopathic effect of LT 910N and LT-YN viruses. The reason for these effects of the exogenous neuraminidase is discussed. PMID- 6271684 TI - Studies of immune responses during recovery from Sindbis virus encephalitis in selectively reconstituted, thymectomized, lethally irradiated mice. AB - Recovery from acute Sindbis virus encephalitis was studied in selectively reconstituted, thymectomized, irradiated mice. Intact mice cleared virus from the brain by day 10 and developed inflammation along with immunoglobulin M (IgM) and IgG antibodies 4 to 6 days after infection. Unreconstituted mice died by day 11, with virus still present but decreasing, no detectable antibody, and no evidence of inflammation. Mice reconstituted with bone marrow cells alone produced only IgM antibody and no inflammation, but cleared virus by day 14. Mice reconstituted with sensitized T cells alone cleared virus by day 10, produced very small amounts of antibody, and developed a prompt inflammatory response. PMID- 6271685 TI - Stimulation of Clostridium perfringens enterotoxin formation by caffeine and theobromine. AB - In the presence of 100 micrograms of caffeine per ml or 200 micrograms of theobromine per ml, sporulation of Clostridium perfringens NCTC 8679 rose from less than 1 to 80 or 85%. Enterotoxin concentration increased from undetectable levels to 450 micrograms/mg of cell extract protein. Heat-resistant spore levels increased from less than 1,000 to between 1 X 10(7) and 2 X 10(7)/ml. These effects were partially reversible by the addition of adenosine or thymidine. In the case of NCTC 8238, caffeine and theobromine caused a three- to fourfold increase in the percentages of cells possessing refractile spores and a similar increase in enterotoxin concentration. Heat-resistant spore levels, however, were unaffected. Inosine was ineffective in promoting sporulation in NCTC 8679. PMID- 6271686 TI - In vivo reactivation of herpes simplex virus in rabbit trigeminal ganglia: electrode model. AB - The rabbit provides an excellent model for the study of ocular herpes because herpetic keratitis in the rabbit eye resembles human disease in its clinical features and in its propensity for spontaneous recurrence. This paper presents a method for the electrical induction of multiple episodes of in vivo reactivation of latent HSV-1 infection with peripheral shedding of virus. Physiological levels of current delivered via an electrode implanted over the trigeminal ganglion of latently infected animals has enabled us to modify and synchronize virus shedding in preocular tear film and to cause multiple episodes of reactivation in a single animal. For this reason, the model is well suited for antiviral efficacy testing and provides an excellent opportunity for investigation of virus-host cell interactions in latent and recurring herpetic disease. PMID- 6271687 TI - Shigella sonnei plasmids: evidence that a large plasmid is necessary for virulence. AB - Virulent form I Shigella sonnei strains contain a 120-megadalton plasmid that is absent in their form II derivatives, which are always avirulent and devoid of O side chains. In the present study, 165 biochemical and antibiotic traits were assessed, but no experimentally useful phenotype could be associated with this large form I plasmid. Therefore, the form I plasmids of several S. sonnei strains were tagged with the antibiotic resistance transposons Tn3, Tn5, or Tn10. Transposon-tagged form I plasmids were not self-transmissible, but could be mobilized by the plasmid R386. Form II S. sonnei transconjugants for the form I plasmid acquired both virulence and the ability to synthesize form I antigen, establishing that these properties are plasmid mediated. Further studies indicate that this 120-megadalton form I plasmid is physically unstable in any of several host bacteria and suggest that it is a member of the FI incompatibility group. Also, two commonly observed, small plasmids of S. sonnei, of 3.2 and 3.9 megadaltons, were shown to encode either colicin E1 production or resistance to streptomycin and sulfonamide, respectively. PMID- 6271688 TI - Evidence that the spontaneous blastogenesis of lymphocytes from bovine leukemia virus-infected cattle is viral antigen specific. AB - Cattle lymphocytes cultured for 3 days were found to spontaneously incorporate thymidine (3STI). Under optimal conditions of culture, the median magnitude of 3STI activity in lymphocytes from bovine leukemia virus (BLV)-infected cattle was higher than that of BLV-free cattle, but the ranges of the values overlapped. However, the 3STI activity of most BLV-infected cattle was specifically inhibited by serum containing BLV antibodies, whereas the 3STI activity of BLV-free cattle was not. The 3STI inhibitor copurified with immunoglobulin, and its activity could be absorbed with BLV. Rabbit anti-BLV serum inhibited 3STI, but rabbit anti BLV p25 did not. These results indicate that BLV infection induces or expands a BLV-specific lymphocyte population. Spontaneous blastogenesis may be indicative of an immune response which controls virus spread. PMID- 6271690 TI - Pharmacolinetic and pharmacodynamic data analysis of theophylline for three different drug forms. AB - Theophylline, which is widely used in asthmatics, should be maintained within narrow plasma limits to achieve maximum therapeutic benefit with minimum adverse effects. For the evaluation of drug input schemes it is essential to know the pharmacokinetic parameters and their variation for different dosage forms. Apart from absorption and elimination rates, an appropriate dosage schedule is determined by the extent of bioavailability. The objectives of the study presented here are the assessment of bioavailability, the estimation of pharmacokinetic parameters, and the investigation of the relationship between plasma drug levels and cardiac side effects. Additionally, it was checked whether drug levels in saliva could be used for monitoring drug levels in plasma. The time course of theophylline in plasma and saliva was measured in twelve healthy volunteers on different drug forms: intravenous injection as well as oral application of a fast and a slow release form of theophylline. Pharmacokinetic parameters were estimated using a three-compartment model for the distribution of theophylline. The extent of bioavailability was estimated via the area under the curves and by numerical evaluation of the invasion function. By simultaneous measurements of heart rate and c-AMP levels the drug response was monitored. The results show the following: i) The slow and fast release dosage forms do not differ with respect to extent of bioavailability, whereas bioavailability rates are slower for the sustained release form. ii) Apart from the initial phase (less than or equal to 1 h) plasma and saliva drug levels run approximately parallel. However, the mean concentration ratio has a large variance. iii) The time courses of heart rate are closely related to the time courses of drug level. iv) When the sustained release form is used for a long-term therapy, it is possible to evaluate dosage schedules and dosage regimens that reduce cardiac side effects caused by the fast release form. This was shown by computer simulation carried out on the basis of pharmacokinetic parameters that were estimated from the experimental curves. PMID- 6271689 TI - Serotypes and antimicrobial susceptibility of Streptococcus pneumoniae in West Germany. AB - Two-hundred-and-six strains of Streptococcus pneumoniae were isolated in eight centers in West Germany. The prevalent serotypes were: 19, 3, 6, 7, 23 and 15. Seventy-five percent of the strains tested were antigenically identical to the pneumococcal types included in the 14-valent vaccine Pneumovax. Susceptibility testing revealed resistance to tetracycline (11% of the isolates), co-trimoxazole (7%) and chloramphenicol (2%). Seven percent of the isolates were relatively resistant to penicillin (MIC 0.1-1.0 mg/l). PMID- 6271691 TI - The analgesic effect of calcitonin in humans: studies on the role of opioid peptides. AB - The possibility that the analgesic action of calcitonin could be due to variations in beta-endorphin levels in both peripheral blood and cerebrospinal fluid was studied. A total of 40 microgram of synthetic salmon calcitonin was injected i. v. in four male volunteers undergoing minor operations under rachianesthesia. Beta-Endorphin was determined in blood and cerebrospinal fluid samples that were collected before and at regular intervals after calcitonin injection. No statistically significant variation in beta-endorphin levels was observed either in peripheral blood or cerebrospinal fluid. Since the injected calcitonin was free of contaminants (demonstrated by both high pressure liquid chromatography and radioimmunoassay of endorphin made on the injected preparation) and since an increase was observed in the immunoreactive calcitonin in cerebrospinal fluid following calcitonin i. v., self-analgesic action may be hypothesized, even though action through prostaglandins or other systems cannot be excluded. PMID- 6271692 TI - The detection of beta-adrenoceptors on murine lymphocytes. PMID- 6271693 TI - Activation of cyclic AMP-dependent protein kinase activity during LPS stimulation of macrophage tumor cell line, J774.1. PMID- 6271694 TI - The stimulation of rat and mouse macrophages by cyclomunine after in vitro and in vivo administration. PMID- 6271695 TI - Photochemistry of pyrimidine bases as studied by e.s.r. and spin-trapping. AB - The direct photoexcitation of pyrimidine bases in D2O solutions yields free radicals which could be conveniently identified by spin-trapping with 2-methyl-2 nitrosopropane. Most of the radicals formed were attributed to D-addition to one end of the 5,6 double bond. However, orotic acid and iso-orotic acid yielded N(3) centred free radicals, formed by homolytic cleavage of the N-H bond. No indication could be found for a free radical involvement in the photocleavage of cyclobutane-type pyrimidine dimers. PMID- 6271696 TI - Hydrogen-deuterium exchange in gamma-irradiated polycrystalline DL-alanine: a spin-trapping and e.s.r. study. AB - The hydrogen-deuterium exchange reactions in gamma-irradiated DL-alanine in the solid state were investigated by spin-trapping and electron spin resonance (e.s.r) using selectively deuterated DL-alanine. Subsequent to gamma-radiolysis at 30 degrees C, polycrystalline DL-alanine was dissolved in aqueous solutions of 2-methyl-2-nitrosopropane and the extent of H-D exchange of the deamination radicals was followed by e.s.r. After formation of the deamination radicals, four exchange reactions were found to occur between the radicals and the surrounding undamaged molecules. The first reaction, which occurs between the hydrogens of the C-2 carbon of the radicals and those of the methyl groups of the neighbouring molecules, can be followed at room temperature. The three other reactions could be conveniently monitored in gamma-irradiated polycrystalline alanine at 110 degrees C. The first of the other three reactions takes place between the methyl hydrogens of the radicals and the C-2 hydrogens of nearby molecules, while the remaining processes involve exchange between the hydrogen atoms of the amino group and those on the C-2 and C-3 carbon atoms of the deamination radical. PMID- 6271697 TI - 2,2,5,5-tetramethyl-1-pyrrolidinyl-oxy-3-carboxylic acid (PCA) as a potential radiosensitizer iv vivo. PMID- 6271698 TI - Changes in electrophoretic mobility of erythrocytes exposed to ionizing radiations of various LET. PMID- 6271699 TI - Radiation-induced free radicals in solid 5-halouracil derivatives: single crystals of 1-methyl-5-bromouracil. AB - Single crystals of the 5-halogen substituted uracil derivative 1-methyl-5 bromouracil have been irradiated at 77 and 295 K and studied by electron spin resonance spectroscopy. At 295 K a hydrogen abstraction radical (I) from the methyl group was observed, together with a modified 5-yl radical (II). The latter species seem to be formed by addition of a non-interacting residue (probably CH3) to C6. At 77 K, radical I is observed together with two bromine-centered species. One of these (III) exhibits only bromine hyperfine interaction (maximum splitting of 515 G) and is assigned to a complexed cation radical. The third 77 K resonance could be due to a beta-bromine radical R'-C5HBr-C6H-R'. A computer program able to calculate powder spectra of radicals with dominant halogen interaction is described. The results are discussed in relation to radiation damage processes in the isomorphous crystals of 1-methylthymine. PMID- 6271700 TI - E.s.r. and spin-trapping studies of dihydropyrimidines. gamma-Radiolysis in the polycrystalline state and U.V. photolysis in aqueous solution. AB - gamma-Radiolysis in the polycrystalline state and U.V. photolysis in aqueous solution at 220 nm of several dihydropyrimidines and their derivatives have been investigated by spin-trapping and electron spin resonance. 2-Methyl-2 nitrosopropane was used as the spin-trap. The spin-adducts of the 6-yl radicals obtained fall into two categories. Those from dihydro-1-methyluracil, dihydro-6 methyluracil, dihydro-1-ethyluracil and dihydro-1-methylcytosine exhibit a beta nitrogen hyperfine coupling constant (alpha beta N) equal to or less than 2.0 G while the ones fom dihydro-orotic acid, dihydrouracil and dihydrothymine showed much larger alpha beta N values (greater than 3.3 G). Dihydrouridine gives radicals characteristic of both the dihydropyrimidine ring and the sugar moiety. The same radicals were obtained by gamma-radiolysis or U.V. photolysis. For all the 5-yl radicals obtained by U.V. photolysis, a direct photoexcitation mechanism is proposed. PMID- 6271701 TI - Comparison of injected and oral polio vaccine for booster immunization during the 1979 polio outbreak in Lancaster County, Pennsylvania. AB - During a 1979 outbreak of poliomyelitis in Lancaster County, Pennsylvania, we investigated the necessity for and the response to booster vaccination of hospital personnel. The immune response of hospital employees who received booster injections of Salk vaccine (n = 38) was compared with that of residents in the surrounding community who received Sabin trivalent OPV boosters (n = 43). Serum neutralizing antibody titers to the three polio serotypes were measured before and after booster immunization. Results indicated that 38% of the subjects in both groups had low initial antibody titers. Salk vaccine was in all circumstances as effective or better than Sabin vaccine in increasing neutralizing IgG antibody titers. PMID- 6271702 TI - Ocular lesions associated with dissemination of type 2 herpes simplex virus from skin infection in newborn rabbits. AB - The subcutaneous inoculation of the backs of New Zealand white rabbits 17 to 34 hr old with 10(3) 50% tissue culture infection dose (TCID50) of type 2 herpes simplex virus (HSV-2) induced cutaneous lesions within 24 hr, foci of disseminated infection in many organs (including the eye) on day 3 and thereafter, and the death of the animals on day 5 with infection of the central nervous system. Infectious HSV-2 could be isolated from the mononuclear cells and plasma of the peripheral blood, indicating the active role of both elements in the dissemination of the virus. Infectious HSV was also recovered from the corresponding sensory ganglia of the skin lesion (the cervicothoracic ganglia) as early as 2 days after the subcutaneous inoculation of the virus. About 40% of the animals developed ocular consisting of retinal folds with or without degenerative changes, Iritis and choroiditis also developed in some eyes. Infectious HSV-2 could be isolated from 33% of the eyes on days 4 and 5. Thus the newborn rabbit may serve as a suitable experimental animal for the study of HSV-2-induced chorioretinitis in the human newborn. PMID- 6271704 TI - Estimation of mouse mammary tumor virus gp52 abundance by protein A assay: a comparison of viral antigen expression on C3H and GR mammary tumor cells. AB - An adherent cell isotopic staphylococcal protein A test (ISPAT) was used to estimate the abundance of mouse mammary tumor virus (MMTV) gp52 cell surface antigen (CSA)on mammary tumor cells. Protein A assays were first utilized, not strictly as a means of antigen detection, but as a means to determine the kinetics of dexamethasone-mediated changes in gp52 cell surface expression. Results indicated increases in gp52 CSA within 4 h of dexamethasone treatment and maximal levels of antigen expression within 12-24 h after treatment. Comparison of gp52 determinants on dexamethasone-stimulated mammary tumor cells demonstrated a greater abundance on C3H Mm5mt/cl than on GR-MMTV cultures. Parallel antigen assays with gp52 and Rauscher murine leukemia virus (R-MuLV) antisera demonstrated that GR-MMTV cells expressed fewer C-type determinants and thus a more preferential expression of gp52 determinants than other cell lines tested. The gp52/R-MuLV binding ratio was only 2.4:1 for C3H cultures in contrast to 5.7:1 for GR cultures. In addition, a comparison of gp52 expression on viral producer and nonproducer cells provided a quantitative estimate of the extent of expression which occurred in a retrovirus nonproducer culture. Results of ISPAT demonstrated that 75% of the gp52 detected on producer cells was present on nonproducer cultures. Comparison of the expression of gp52 CSA and the release of gp 52 into culture fluids during hormone treatments demonstrated that both assays (ISPAT and gp 52 radioimmunoassay) detect and quantitate coordinate changes in the expression and release of MMTV gp52. In antibody excess, protein A assays provided quantitative estimates of CSA abundance not offered by alternative methods of CSA detection. PMID- 6271703 TI - Immunology of pyelonephritis in the primate model. II. Effect on immunosuppression. AB - Nonobstructive pyelonephritis was produced in the rhesus monkey (Macaca mulatta) by means of retrograde inoculation of Escherichia coli to the point of pyelotubular backflow. The effects of treatment with cyclophosphamide or azathioprine were determined. Commensal renal viruses were not activated by the immunosuppression, and thus did not complicate our results. Both cyclophosphamide and azathioprine prolonged the bacteriuria and produced more severe pathology. Cyclophosphamide decreased the leukocytic response and partically suppressed the antibody response. The increased amount of acute pyelonecytic response and partially suppressed the antibody response. The increased amount of acute pyelonephritic lesions after treatment with this drug suggest that the antibody and inflammatory responses may be important protective mechanisms, particularly regarding prevention of abscesses. In contrast, azathioprine did not decrease the leukocytosis nor the antibody responses, but resulted in decreased in vitro responsiveness of lymphocytes to mitogens. The increased severity of chronic pyelonephritic lesions after azathioprine treatment suggests that the cellular immune response also may be an important protective mechanism during late stages of the disease. The results thus indicate that the immune response is protective and is not directly responsible for the chronic scarring of pyelonephritis. PMID- 6271705 TI - Argentine hemorrhagic fever: a biologic marker. AB - Mortality rates and viral replication in blood and brains of Wistar rats between 6 h and 26 days of age inoculated with two strains of Junin virus of different virulence were compared. Viral growth curves in brains showed no differences between strains. However, differences in mortality rates were significant among rats between 1 and 3 days of age. When the intracerebral (i.c.) route was used, high mortality rates were induced by the attenuated XJC13 strain and low mortality rates were induced by the pathogenic XJ strain. On the other hand, when the intraperitoneal or subcutaneous route was used, mortality rates were reversed: low for the attenuated strain and high for the pathogenic one. The use of different doses of each virus and the application of various routes of inoculation in 2-day-old rats showed that 10(3) TCID50 by the i.c. route resulted in the greatest difference in mortality rates. PMID- 6271706 TI - Evidence for papillomavirus genus-specific antigens and DNA in laryngeal papilloma. AB - Four juvenile laryngeal papillomas (JLPs) were examined for papillomavirus genus specific structural antigens and polynucleotide sequences employing reagents derived from bovine papillomavirus type 1 (BPV-1). Three of four JLPs contained intranuclear antigens in the granular layer of differentiating epithelial cells of formalin-fixed, paraffin-embedded papillomas assayed by the peroxidase antiperoxidase reaction using an antiserum prepared against purified detergent disrupted BPV-1. Examination of these JLPs for viral sequences indicated two tumor DNA preparations contained polynucleotide sequences which hybridized to an in vitro labeled BPV-1 DNA probe under nonstringent conditions. These sequences were detectable after gel electrophoresis and immobilization of restriction endonuclease-digested JLP DNAs onto nitrocellulose membranes. Specific hybridization of probe to digested JLP DNA was visualized as bands in autoradiograms coincidental with open circular and linearized papillomavirus DNA. Hybridization was extensively reduced when JLP DNA preparations were prehybridized to an unlabeled papillomavirus DNA heterologous to the probe. These results provide direct evidence for the association of a papillomavirus in JLP. PMID- 6271707 TI - Detection of specific IgA antibodies in serum of kidney transplant patients with recurrent cytomegalovirus infection. AB - 59 sera of 10 immunosuppressed renal allograft recipients who experienced recurrent cytomegalovirus (CMV) infection were analyzed by enzyme-linked immunosorbent assay (ELISA) for CMV IgA antibodies and by the complement-fixation (CF) test. A significant rise of CF titer was evident 4-53 weeks post transplantation. 9 patients produced CMV IgA in high titers at about the time of CF antibody rise was observed. 1 patient did not produce IgA antibodies to CMV. In 3 of the 9 patients, specific CMV IgA antibodies were detected before the rise if CF titer was demonstrated. CMV IgA antibodies were found to persist for as long as 6 weeks post-transplantation. The potential application of ELISA detection of CMV-specific IgA antibodies as an early indication of CMV infection in kidney transplant patients is discussed. PMID- 6271708 TI - Promotion of comprehensive care to the psychiatric aftercare client. PMID- 6271709 TI - The enhancing effect of cytochalasin A on concanavalin A-induced superoxide anion release from polymorphonuclear leukocytes. PMID- 6271710 TI - A cytophotometric measurement of DNA in murine hepatocytic nuclei during cytomegalovirus infection. AB - The amount of Feulgen-DNA in the nuclei of normal murine hepatocytes as well as those from livers on the third and fifth day after lethal, cytomegalovirus infection was assessed by scanning cytophotometry. The technique enabled the measurement of the increasing content of DNA in hepatocytic nuclei during the course of the disease. The results suggest that although some of this increase may be due to cellular DNA synthesis, most of it is due to viral DNA replication. Lastly, the megalohepatocytes which characterise this disease reflect viral replication in predominantly diploid hepatocytes. PMID- 6271711 TI - Factors determining temporal pattern of isobaric supersaturation. AB - It is possible to produce a transient supersaturation or undersaturation in tissues and blood by sequentially breathing gases with different equilibration rates. If the ambient gas pressure is sufficiently high, the induced supersaturation can produce vascular bubbles. By means of the classical perfusion dependent model of inert gas elimination, which assumes that the effects of diffusion are minimal, the magnitude of the total inert gas pressure can be predicted. If, however, the effects of diffusion cannot be ignored, the supersaturation could be substantially larger. This paper estimates the effects of diffusion in a Krogh cylinder on the supersaturation produced by suddenly changing the inert gas partial pressure in the blood. The results of these estimates indicate that diffusion plays a role in this transient supersaturation only in long Krogh cylinders with high blood flows. The effects of diffusion are further reduced by the finite time necessary to switch the inert gases in arterial blood. The conclusions are supported by experiments that measure vascular bubble production after a switch of the inert portion of the inspired gas. These experiments further show that the formation of vascular bubbles after such a switch cannot be entirely explained by the different diffusion constants of the gases used. PMID- 6271712 TI - Acute pulmonary hemodynamic effects of intravenous copper sulfate: role of alpha adrenergic system. AB - We investigated the acute pulmonary hemodynamic effects of intravenous copper sulfate (CuSO4) infusion and its mechanism of action in six groups of conscious sheep (total 40). After 300 mg CuSO4 alone, mean pulmonary artery pressure (Ppa) increased from 10.3 to 22.5 Torr and pulmonary artery wedge pressure (Ppaw) from 3.5 to 7.6 Torr, whereas systemic arterial pressure (Psa) increased from 95 to 102 Torr. Cardiac output (Qp) decreased from 4.7 to 3.3 l/min. Pulmonary vascular resistance (PVR) and systemic vascular resistance (SVR) increased to 320 and 160% of base line, respectively. The hemodynamic changes correlated well with serum copper, which increased from a base-line value of 0.12 to 3.5 mg/dl after the CuSO4. Serum dopamine beta-hydroxylase increased from 3.2 U/l before CuSO4 injection to 5.7 after its administration, signifying activation of adrenergic nervous system. H1-histamine receptor blockade with chlorpheniramine failed to prevent the effects of CuSO4. Pretreatment with methysergide, a serotonin antagonist, partially attenuated the effects of CuSO4. Phenoxybenzamine, an alpha adrenergic receptor blocker, and 6-hydroxydopamine, a catecholamine depleting agent, completely blocked the effects of CuSO4. beta-Adrenergic receptor blockade with propranolol enhanced the effects of CuSO4. We conclude, that, in conscious sheep, acute infusion of CuSO4 caused a marked reversible increase in PVR with a slight transient increase in SVR, and this pulmonary hypertension was produced by stimulation of the alpha-adrenergic nervous system. PMID- 6271714 TI - ACTH and cortisol responses to hypoxia in dogs. AB - To determine the role of endogenous ACTH and hyperventilation in the adrenocortical response to hypoxia, pentobarbital-anesthetized dogs equipped with left adrenal venous cannulas for measurement of cortisol secretion rate (CSR) and arterial cannulas for measurement of plasma ACTH were exposed to 20 min of normoxia (group I), spontaneous ventilation, normocapnic hypoxic hypoxia (group II), controlled ventilation, normocapnic hypoxic hypoxia (group III), controlled ventilation, normocapnic carbon monoxide hypoxia (group IV), or hypoxic hypoxia with elevated carboxyhemoglobin (group V). Group I showed no change in ACTH and CSR. Groups II and III greatly increased CSR whereas only group III increased ACTH significantly. Group IV greatly increased ACTH whereas CSR increased but less than group III. Group V showed a significant increase in ACTH but no significant CSR response. In addition, 5 U of ACTh were infused in several animals from groups I, III, and IV. Exogenous ACTH caused increases in CSR that were larger in group I than groups III and IV. The data are consistent with the hypothesis that ACTH in arterial blood is not the sole controller of CSR during hypoxic stress. PMID- 6271713 TI - Autonomic mechanisms of training bradycardia: beta-adrenergic receptors in humans. AB - To address the autonomic mechanisms underlying the bradycardia of physical training in human subjects, we performed a cross-sectional study comparing the heart-rate responses to graded doses of isoproterenol in 7 elite marathon runners and 7 age-matched controls, and a longitudinal study in 12 normal volunteers of the effects of 6 wk of intense physical training on lymphocyte beta-adrenergic receptors identified by l-[3H]dihydroalprenolol. We observed no significant differences between marathoners and controls in the dose of isoproterenol that produced a 25-beat/min increment in heart rate, either in the absence (1.9 +/- 0.6 vs. 2.5 +/- 0.6 microgram; P, 0.509) or in the presence of cholinergic blockade (4.4 +/- 1.3 vs. 3.1 +/- 0.4 microgram: P, 0.320). Likewise, we observed no effects of physical training on lymphocyte beta-adrenergic receptors in terms of receptors number (53 +/- 11 vs. 56 +/- 10 fmol/mg protein) or receptor affinity (Kd 4.0 +/- 0.7 vs. 3.6 +/- 0.7 nM) (P, 0.9178). Although our data cannot exclude reduced chronotropic sensitivity to catecholamines as contributing to lowered heart rate in some highly conditioned individuals, these results are consistent with the hypothesis that altered neuronal input to the sinus node is usually a more important mechanism of training bradycardia. PMID- 6271716 TI - Synthesis and structure-activity relationships of 7 beta-[2-(2-aminothiazol-4 yl)acetamido]cephalosporin derivatives. V. Synthesis and antibacterial activity of 7 beta-[2-(2-aminothiazol-4-yl)-2-methoxyiminoacetamido]-cephalosporin derivates and related compounds. AB - In order to improve the antibacterial activity of 7 beta-[2-(2-aminothiazol-4 yl)acetamido]-cephalosporins new derivatives having a methoxyimino moiety in the 7-acyl side chain and related compounds were synthesized. Of these, 7 beta-[2-(2 aminothiazol-4-yl)-(Z)-2-methoxyiminoacetamido]-cephalosporins were found to possess excellent activity against a variety of Gram-positive and Gram-negative bacteria including beta-lactamase-producing strains. An extensive study of structure-activity relationships led to the selection of 7 beta-[2-(2 aminothiazol-4-yl)-(Z)-2-methoxyiminoacetamido]-3-[(1-methyl-1H-tetrazol-5-yl) thiomethyl]-ceph-3-em-4-carboxylic acid, SCE-1365, for further biological and clinical evaluation. PMID- 6271717 TI - Synthesis and structure-activity relationships of 7 beta-[2-(2-aminothiazol-4 yl)acetamido]cephalosporin derivatives. VI. Alternative syntheses of 7 beta-[2-(2 aminothiazol-4-yl)-(Z)-2-methoxyiminoacetamido]cephalosporin derivatives. PMID- 6271715 TI - Respiratory depression produced by activation of GABA receptors in hindbrain of cat. AB - Respiratory responses to activation of gamma-aminobutyric acid (GABA) receptors in the hindbrain were measured in chloralose-anesthetized cats using a Fleisch pneumotachograph. GABA receptors were activated by intracisternal injections of muscimol and GABA. Muscimol (0.05--6.65 micrograms) administered to seven animals caused a depression of respiratory activity with apnea occurring in each animal. Before apnea occurred, a decrease in tidal volume was observed (from 25.7 +/- 0.9 to 14.7 +/- 1.1 ml). Respiratory rate and inspiratory and expiratory durations were unchanged. GABA (0.05--12.15 mg) administered to five animals produced the same effect as muscimol on respiratory activity. Apnea produced by both agents was reversed by intracisternal administration of the GABA-receptor antagonist drug, bicuculline. Administration of bicuculline to four naive animals increased tidal volume (from 31.3 +/- 1.7 to 36.5 +/- 0.7 ml) but had no effect on either respiratory rate or inspiratory duration. These results indicate that activation of GABA receptors causes respiratory depression and suggest that GABA may be an important neurotransmitter in CNS neural pathways involved in regulating respiratory activity. PMID- 6271718 TI - Modification of aminoglycoside antibiotics by clinical isolates of Streptococcus faecalis. PMID- 6271719 TI - Relevance of in vitro antibacterial activities of cefotiam and cefazolin to their therapeutic effects on experimental pneumonia caused by Klebsiella pneumoniae DT S in mice. AB - The MICs of cefotiam and cefazolin against K. pneumoniae DT-S were unaffected by the inoculum size and were 0.1 and 1.56 micrograms/ml, respectively. Bactericidal and bacteriolytic activities of the cephalosporins were more potent in bacterial concentrations of 10(7) colony-forming units (CFU)/ml than in concentrations of 10(8) CFU/ml. Both activities of cefotiam were more markedly influenced by bacterial concentrations than those of cefazolin. Therapeutic activity of cefotiam was about 9 approximately 15 times as potent as that of cefazolin in experimental pneumonia caused by K. pneumoniae DT-S in mice, and this finding was in accordance with the ratio of in vitro antibacterial activities of the two cephalosporins as judged by the MICs or the bactericidal and bacteriolytic activities in bacterial suspension of 10(7) CFU/ml. The range of concentrations of cefotiam which induced cell filamentation in vitro, was wider than that of cefazolin. This difference, however, was not reflected on the therapeutic activities of the two cephalosporins in the model infection. In the pneumonic lungs, definite therapeutic doses of both cephalosporins (80 mg of cefotiam per kg and 640 mg of cefazolin per kg) produced mainly bacteriolysis of the challenge organisms. PMID- 6271720 TI - Effect of nutritional status on swine adipose tissue lipolytic activities. AB - Young swine (28 days of age) were fed an isocaloric and isonitrogenous diet with either a high fat or a low fat content for 3 to 4 weeks. The adipose tissue lipolytic rate was higher in the group fed the high fat diet. However, there was no effect of diet on the activities of several of the enzymes controlling the lipolytic process, i.e., adenylate cyclase, phosphodiesterase and hormone sensitive lipase. No effect of diet on the activity of lipoprotein lipase was detected. Fasting for 72 hr, but not for 24 or 48 hr, caused an increase in the lipolytic rate. There was also a decrease in cell size after a 72-hr fast (P greater than .05) such that the increased rate was not significant when the data were expressed on a cell basis. Inexplicable transient changes in adenylate cyclase activity, as well as a decrease in the activity of the low affinity phosphodiesterase (doubtful physiological significance), were detected during starvation. Starvation depressed the adipose tissue lipoprotein lipase activity but had no effect on the hormone-sensitive lipase activity. PMID- 6271721 TI - A study of rapid and simplified confirmatory tests for Clostridium perfringens. PMID- 6271722 TI - Susceptibility to mecillinam and other antibiotics of 28 O-serotypes of Yersinia enterocolitica. PMID- 6271723 TI - Pharmacokinetics of ketoconazole in normal subjects. PMID- 6271724 TI - Inhibition of clonal growth of Giardia lamblia and Entamoeba histolytica by metronidazole, quinacrine, and other antimicrobial agents. PMID- 6271725 TI - Comparative activity and beta-lactamase stability of carbenicillin, ticarcillin, azlocillin, mezlocillin and cefotaxime. PMID- 6271726 TI - Comparison of the beta-lactamase stability and the in-vitro activity of cefoperazone, cefotaxime, cefsulodin, ceftazidime, moxalactam and ceftriaxone against Pseudomonas aeruginosa. PMID- 6271727 TI - Antibacteroides and beta-lactamase inhibitory activities of moxalactam. PMID- 6271728 TI - Synthesis of teichoic acid by Bacillus subtilis protoplasts. AB - Protoplasts of Bacillus subtilis W23 readily synthesized ribitol teichoic acid from nucleotide precursors in the surrounding medium. With cytidine diphosphate ribitol they made poly(ribitol phosphate), presumably attached to lipoteichoic acid carrier; when cytidine diphosphate-glycerol and uridine diphosphate-N acetylglucosamine were also present a 10-fold increase in the rate of polymer synthesis occurred, and the product contained both the main chain and the linkage unit. Synthesis was inhibited by trypsin or p-chloromercuribenzenesulfonate in the medium, and we concluded that it occurred at the outer surface of the membrane. During synthesis, which was also achieved readily by whole cells after a brief period of wall lysis, the cytidine phosphate portion of the nucleotide precursors did not pass through the membrane. No evidence could be obtained for a transphosphorylation mechanism for the translocation process. It is suggested that reaction with exogenous substrates was due to temporary exposure of a protein component of the enzyme complex at the outer surface of the membrane during the normal biosynthetic cycle. PMID- 6271729 TI - Molecular cloning of gene xylS of the TOL plasmid: evidence for positive regulation of the xylDEGF operon by xylS. AB - The xylDEGF operon and the regulatory gene xylS of the TOL plasmid found in Pseudomonas putida mt-2 were cloned onto Escherichia coli vector plasmids. A 9.5 kilobase fragment, derived from the TOL segment of pTN2 deoxyribonucleic acid, carried the xyl genes D, E, G, and F, which encode toluate oxygenase, catechol 2,3-oxygenase, 2-hydroxymuconic semialdehyde dehydrogenase, and 2-hydroxymuconic semialdehyde hydrolase, respectively. The enzymes were noninducible unless a 3 kilobase PstI fragment, derived also from the TOL segment, was provided in either cis or trans. The PstI fragment appeared to contain the regulatory gene xylS, which produced a positive regulator. The regulator was activated by m-toluate or benzoate, but not by m-xylene or m-methylbenzyl alcohol. the map positions of xylG and xylF were also determined. PMID- 6271730 TI - New nalidixic acid resistance mutations related to deoxyribonucleic acid gyrase activity. AB - In Escherichia coli K-12 mutants which had a new nalidixic acid resistance mutation at about 82 min on the chromosome map, cell growth was resistant to or hypersusceptible to nalidixic acid, oxolinic acid, piromidic acid, pipemidic acid, and novobiocin. Deoxyribonucleic acid gyrase activity as tested by supercoiling of lambda phage deoxyribonucleic acid inside the mutants was similarly resistant or hypersusceptible to the compounds. The drug concentrations required for gyrase inhibition were much higher than those for cell growth inhibition but similar to those for inhibition of lambda phage multiplication. Transduction analysis with lambda phages carrying the chromosomal fragment of the tnaA-gyrB region suggested that one of the mutations, nal-31, was located on the gyrB gene. PMID- 6271731 TI - Lipid alterations in cell envelopes of polymyxin-resistant Pseudomonas aeruginosa isolates. AB - The lipid composition of cells of Pseudomonas aeruginosa strains resistant to polymyxin was compared with the lipid composition of cells of polymyxin-sensitive strains as to their content of readily extractable lipids (RELs), acid extractable lipids, the fatty acid composition of RELs, and the contents of various phospholipids in the REL fraction. The polymyxin-resistant strains had an increased content of RELs, but a decreased phospholipid content. The REL fraction from the polymyxin-resistant strains had an increased content of unsaturated fatty acids accompanied by a decreased content of cyclopropane fatty acids as compared with the fatty acid composition of RELs from polymyxin-sensitive strains. The phosphatidylethanolamine content was greatly reduced in the polymyxin-resistant strains, whereas the content of an unidentified lipid, thought to be a neutral lipid lacking either a phosphate, free amino, or choline moiety, was greatly increased. Cell envelopes of the polymyxin-resistant strains contained reduced concentrations of Mg2+ and Ca2+ as compared with the cell envelopes of polymyxin-sensitive strains. It appears that polymyxin resistance in these strains is associated with a significant alteration in the lipid composition and divalent cation content of the cell envelope. PMID- 6271732 TI - Cloning of immunity and structural genes for colicin V. AB - The colicin V immunity and structural genes of plasmid pColV-B188 were cloned into the vectors pMB9, pBR322, and pMK16. Both genes are closely linked and can be isolated on a 900-base-pair deoxyribonucleic acid fragment. Insertion of the transposon Tn5 into this cloned sequence led to the construction of a mutant plasmid which conferred colicin V immunity, but not the ability to produce this colicin. Analysis of the products determined by these cloned genes in cells has led to the conclusion that the polypeptide involved in immunity has a molecular weight of about 6,500, whereas the colicin has a molecular weight of approximately 4,000. PMID- 6271733 TI - Effects of growth substrate and respiratory chain composition on bioenergetics in Acinetobacter sp. strain HO1-N. AB - The relationship between respiratory chain composition and efficiency of coupling phosphorylation to electron transport was examined in Acinetobacter sp. strain HO1-N. Cells containing only cytochrome o as a terminal oxidase displayed the same stoichiometries of adenosine 5'-triphosphate synthesis and proton extrusion as cells which contained both cytochromes o and d as terminal oxidases. In addition, CO inhibition and photo-relief of cytochromes o or d did not alter the efficiency of energy coupling. These findings indicate that adenosine 5' triphosphate synthesis is coupled to electron transport through both cytochromes o and d in Acinetobacter. PMID- 6271734 TI - Regulation of fatty acid degradation in Escherichia coli: analysis by operon fusion. AB - Fusion of the lacZ gene coding for beta-galactosidase to the fadA,B and fadE operons was accomplished by using the phage Mu d (Apr lac). In such fusion strains, beta-galactosidase was induced by long-chain fatty acids and repressed by glucose, as is the normal pattern of control for the enzymes of the fad regulon. The level of induction seen was approximately 10-fold for both the fadA and fadE operons. These results demonstrate that the previously observed regulation of both the fadA and fadE operons is at the transcriptional level. When an insertion mutation in the fadR (repressor) gene was introduced into the fusion strains, beta-galactosidase was produced constitutively. A series of fatty acids of different chain lengths were tested as inducers. Acids of chain lengths of 10 carbon atoms or less failed to induce, those of 12 carbon atoms induced partly, and those of 14 or more carbon atoms induced fully. Imidazole was found to counteract the glucose repression of the fadA operon as recently demonstrated for the ara operon. PMID- 6271736 TI - Induction of tryptophanase in short cells and swarm cells of Proteus vulgaris. AB - Tryptophanase was noninducible in swarm cells of Proteus vulgaris despite transport of the inducer tryptophan. Further, cyclic AMP, which stimulated increased levels of tryptophanase in short cells, had no effect on swarm cells. PMID- 6271735 TI - Characterization of a macrolide, lincosamide, and streptogramin resistance plasmid in Staphylococcus epidermidis. AB - A strain of Staphylococcus epidermidis was transduced to erythromycin resistance, and all of the transductants exhibited the macrolide, lincosamide, streptogramin B resistance phenotype. Curing and antibiotic disk studies also indicated that these resistances were controlled by a single plasmid determinant and were constitutive. Agarose gel electrophoresis of plasmid deoxyribonucleic acid (DNA) from donor, cured, and transduced strains showed that a single plasmid was responsible. This plasmid, designated pNE131, was examined for sequence homology to two other plasmids, pE194 and p1258, from Staphylococcus aureus, which also code for erythromycin resistance. DNA from plasmids pNE131 and pE194 hybridized with one another, but no extensive homology to pI258 with either pNE131 or pE194 was found. Restriction endonuclease digests of pNE131 and pE194 showed no common fragments. However, sequence homology was localized to the nucleotides in pE194 that code for the 29,000-dalton protein responsible for erythromycin resistance. pNE131 was calculated to have 2,220 base pairs and is the smallest naturally occurring plasmid with a known function yet reported in S. epidermidis. PMID- 6271737 TI - Suppression of superprecipitation of contractile proteins from chicken gizzard muscle by preincubation in the presence of adenosine triphosphate without Ca2+. AB - Superprecipitation of reconstituted actomyosin composed of smooth muscle myosin, skeletal muscle actin and smooth muscle native tropomyosin was studied. When the actomyosin solution was preincubated in the presence of ATP and the absence of Ca2+, or in the relaxed state, superprecipitation was markedly suppressed. The extent of suppression was correlated with the inhibition of the phosphorylation of the 20,000-dalton light chain of smooth muscle myosin. This is consistent with the theory that the interaction of smooth muscle actomyosin is regulated by the phosphorylation of myosin light chain through a system of myosin light chain kinase and phosphatase. However, further studies showed that the myosin light chain kinase and phosphatase system could not explain the present suppression of superprecipitation, even if a cyclic AMP-dependent protein kinase system was also involved. A new regulatory factor should be taken into account in the regulation of smooth muscle actomyosin interaction. PMID- 6271738 TI - Binding of calcium to lysozyme and its derivatives. AB - Bindings of calcium to lysozyme and its derivatives were studied by UV difference spectroscopy at various pH's. The binding constant was ca. 40 m-1 at around neutral pH. The binding caused proton release from lysozyme and did not inhibit the binding of tri-N-acetylglucosamine to lysozyme. In the presence of 0.2 M Ca2+, lysozyme showed 26% of the activity of the free enzyme toward hexa-N acetylglucosamine but the cleavage pattern was similar to that of the free enzyme. Thus, calcium was predicted to bind near the catalytic carboxyls to cause inhibition of lysozyme activity. It was found from the results of protease digestion that calcium binding shifted the native-denatured transition in lysozyme toward the native state. PMID- 6271740 TI - Presence of an inactive protein immunologically analogous to cytochrome c oxidase in the inner membrane of sweet potato root mitochondria. AB - A protein, which was immunoreactive to antibody against cytochrome c oxidase, was found in the mitochondrial membrane fraction of sweet potato root tissue. The protein was associated relatively weakly with the mitochondrial inner membrane as compared with cytochrome c oxidase. It exerted no cytochrome c oxidase activity and contained no heme a. The protein was purified by phenyl-Sepharose column chromatography and polyacrylamide gel electrophoresis. The molecular weight of its polypeptide chain was 57,000. In addition, the protein decreased during aging of tissue slices. It is therefore not improbable that the protein is a precursor of cytochrome c oxidase composed of only the subunits of cytoplasmic origin, since aging of tissue slices has been shown to result in an increase in the enzyme activity which is inhibited by chloramphenicol but not by cycloheximide. PMID- 6271739 TI - Mechanism of increase in cytochrome c oxidase activity in sweet potato root tissue during aging of slices. AB - The mechanism of an increase in cytochrome c oxidase [EC 1.9.3.1] activity during aging of sliced sweet potato root tissue was investigated with antibiotics and antibody to the purified enzyme. 1. The increase in cytochrome c oxidase activity was inhibited by chloramphenicol but not by cycloheximide. 2. Cytochrome c oxidase purified from wounded tissue was identical with that from intact tissue as judged by the subunit composition, sedimentation velocity, absorption spectrum, antigenicity, and activity per heme a. 3. An increase in the amount of cytochrome c oxidase protein took place during aging of slices. 4. Sweet potato cytochrome c oxidase consists of five subunits. When slices were aged in the presence of [3H]leucine, the three larger subunits (I, II, and III) of cytochrome c oxidase were labeled, while no radioactivity was incorporated into the other two subunits, IV and V. The results indicate that the increase in cytochrome c oxidase activity is due to an increase in the amount of the enzyme protein. We propose that excess amounts of subunits derived from the cytoplasm of the enzyme are present in intact tissue and are assembled with subunits of mitochondrial origin to form the holoenzyme after wounding of tissue. PMID- 6271741 TI - An affinity adsorbent, 5'-adenylate-aminohexyl-sepharose. I. Purification and properties of two forms of RNase U2. AB - An affinity adsorbent, 5'-adenylate-aminohexyl-Sepharose 4B, was prepared by the periodate oxidation of AMp followed by coupling and condensation with amino-hexyl Sepharose 4B. RNase U2, a purine-specific RNase, was specifically bound to this adsorbent at pH 4.5 and eluted critically at pH 5.9 in the presence of 1 M NaCl, corresponding to the pH dependence of the binding of 2'-AMP to RNase U2. By using this affinity chromatography as a main tool, a simplified and effective purification method for RNase U2 was established with a high yield of 58%. Another form of RNase U2 with low specific activity, named RNase U2-B, was eluted at a slightly higher pH from this adsorbent. RNase U2-B was indistinguishable from the original enzyme (RNase U2-A) in base specificity, affinity for ApA, molecular weight and amino acid composition, but was clearly different in specific activity, molecular activity for ApA, isoelectric point and conformation of molecule. This affinity adsorbent is also effective for the detection or isolation of small amounts of base-specific RNases in crude cell extract. PMID- 6271742 TI - An affinity adsorbent, 5'-adenylate-aminohexyl-sepharose. II. Purification and characterization of multi-forms of RNase T2. AB - RNase T2 bound to an affinity adsorbent, 5'-adenylate-aminohexyl-Sepharose 4B, specifically at pH 4.5. The colorless enzyme was eluted only by the simultaneous addition of 2'(3')-AMP (1 mM) and NaCl (greater than 1 M) at pH 4.5. By applying this affinity chromatography to the purification of RNase T2, pure enzyme with a specific activity of 60 was obtained in only four steps and the yield was about 10 times higher than that of the previous purification method. This enzyme preparation was found to be heterogeneous in molecular weight and was separated into two fractions on Sephadex G-75 gel filtration. As the smaller enzyme with a molecular weight of 36,000 was identical with RNase T2 in every property examined, we tentatively designated the larger one with an apparent molecular weight of 80,000 as high molecular weight RNase T2 (RNase T2-L). RNase T2-L was still heterogeneous and was separated into five fractions, RNases T2-L 1-5, by repeated Sephadex G-150 gel filtration. The amino acid and carbohydrate analyses revealed that each of these fractions has a protein moiety in common with RNase T2 and the heterogeneities were due to the carbohydrate content, mainly galactose content. PMID- 6271743 TI - Crystallization of phosphatidylserine bilayers induced by lithium. AB - Utilizing differential scanning calorimetry and x-ray diffraction, 1,2 dimyristoyl-L-glycero-3-phospho-L-serine (DMPS) was shown to form hydrated bilayer membrane structures exhibiting a gel leads to liquid crystalline transition at 39 degrees C (delta H = 7.2 kcal/mol). Addition of up to molar concentrations of the alkali halides NaCl, KCl, Rl Cl, and CsCl produced relatively minor changes in DMPS bilayer structure or stability. For example, in the presence of 0.5 M NaCl, the transition temperature (Tc = 42 degrees C) and transition enthalpy (delta H = 7.0 kcal/mol) show only minor changes. In marked contrast, addition of LiCl results in "'crystallization" of the DMPS bilayer membrane structure. At 0.5 M LiCl, the crystalline DMPS exhibits a bilayer gel leads to liquid crystal transition at 89 degrees C accompanied by a high enthalpy change, delta H = 16.0 kcal/mol. Thus, Li+ induces an isothermal crystallization of DMPS bilayers, the hydrocarbon chains adopting a more ordered packing mode than the "hexagonal" arrangement of the gel state. In view of the widespread use of lithium in the treatment of manic-depressive illness, we also raise the possibility that interaction of Li+ with anionic membrane phospholipids could play a role in its pharmacological action. PMID- 6271744 TI - Evidence that viral transforming gene products and epidermal growth factor stimulate phosphorylation of the same cellular protein with similar specificity. AB - Treatment of A-431 human epidermoid carcinoma cells with epidermal growth factor (EGF) was shown to enhance the phosphorylation of a Mr = 34,000 protein. Because the phosphorylation of an analogous protein is enhanced in various cell lines transformed by Rous sarcoma virus (RSV) (Erikson, E., and Erikson, R. L. (1980) Cell 21, 829-836), we characterized the phosphorylation of the A-431 Mr = 34,000 protein under these two conditions in order to determine whether there are common pathways between viral transformation and EGF stimulation. The results of tryptic phosphopeptide mapping and phosphoamino acid analysis showed that the Mr = 34,000 protein was phosphorylated in an identical manner by the EGF-stimulated protein kinase activity and by the protein kinase activity of the RSV transformation specific protein or of its normal cell homolog. Although the specific protein kinase that phosphorylates the Mr = 34,000 protein under conditions of EGF stimulation is not yet identified, these studies demonstrate that at least one consequence of EGF stimulation is identical with one of the consequences of viral transformation. PMID- 6271745 TI - Purified vesicular stomatitis virus contains an enzyme activity that synthesizes cytidylyl (5'-3') guanosine 5'-triphosphate in vitro. AB - An enzyme activity that synthesizes cytidylyl (5'-3') guanosine 5'-triphosphate (pppGpC) in vitro has been identified in purified vesicular stomatitis virus. The activity is discernible after a lag period which is reduced in length with increasing virus concentration. The lag is eliminated by addition of pppGpC or ppGpC which are effective primers and stimulate dinucleotide synthesis linearly. The requirements of the reaction with respect to MgCl2, NaCl, and temperature are similar to those for viral mRNA synthesis in vitro. The activity, together with the viral L and NS proteins, is removed from virions by treatment with 0.8 M NaCl. The particulate fraction from infected cells that contains the transcribing subviral ribonucleoprotein particles also contains the enzyme activity. The corresponding fraction from uninfected cells does not, indicating that the activity is mediated by virus-specific proteins. Possible functions of the dinucleotide in the life cycle of the virus are discussed. PMID- 6271746 TI - Plant prolyl hydroxylase recognizes poly(L-proline) II helix. AB - Substrate specificity of a prolyl hydroxylase from Vinca rosea suspension cultured cells was studied using synthetic oligo(L-proline)s and their t butyloxycarbonyl derivatives (Pron and Boc-Pron; n = 2-8) as peptidyl substrates. All peptides with a residual number of 5 or greater served as substrates in the enzyme reaction at 30 degrees C, after the preincubation of the enzyme and peptides at 0 degrees C prior to addition of cofactors and cosubstrate. Under the same conditions, the hydroxylation of Pro5 reached a plateau within 10 min, but that of Boc-Pro8 and poly(L-proline) increased linearly up to 40 min. If the preincubation temperature was raised to 30 degrees C, only Pro5 among the peptides was unable to serve as a substrate. The optimum temperature of the enzyme was 30 degrees C toward Boc-Pro8 and poly(L-proline) but it decreased to 15 degrees C using Pro5. These data suggest that the enzyme can bind Pro5 only at low temperature. Poly(L-proline) and Boc-Pron (n greater than or equal to 5) in aqueous solution are known to have a left-handed helical structure (poly(L proline) II helix). Moreover, Pro5 was indicated as forming this helix below 10 degrees C. Accordingly, the enzyme recognizes the poly(L-proline) II helix, that is, the secondary structure of a substrate rather than the primary structure. PMID- 6271747 TI - Removal of the surface-bound human choriogonadotropin results in the cessation of hormonal responses in cultured Leydig tumor cells. AB - The relationship between the binding and internalization of human choriogonadotropin and the stimulation of cAMP and steroid production was studied in cultured Leydig tumor cells. It was found that removal of the surface-bound hormone results in a rapid cessation of both cAMP and steroid production. We propose that the surface-bound hormone is responsible for the activation of steroidogenesis and that hormone internalization is involved in the deactivation of this process. PMID- 6271748 TI - Persistent activation of steroidogenesis in adrenocortical cells by photoaffinity labeling of corticotropin receptors. AB - Photolysis of rat adrenocortical cells in the presence of the photoreactive derivative [(2-nitro-5-azidophenylsulfenyl)Trp9]-adrenocorticotropic hormone (2,5 NAPS-ACTH) at 24 degrees C resulted in persistent activation of corticosterone production. The basal rate of steroidogenesis became maximal when photolysis was performed at 24 degrees C but remained the same as that of control cells when irradiation was performed at 0 degrees C. No increase in basal rate was observed with dark controls or cells photolyzed with [(2,4 dinitrophenylsulfenyl)Trp9]ACTH, a photoresistant analog of the hormone. Prephotolyzed 2,5-NAPS-ACTH failed to induce persistent activation. Both ACTH and 2,4-(dinitrophenylsulfenyl)Trp9-ACTH blocked the photo-induced activation of steroidogenesis elicited by 2,5-NAPS-ACTH. Under photolysis conditions which caused the basal rate of steroidogenesis to become maximal, a 3-fold increase in the basal rate of cAMP formation was observed. PMID- 6271749 TI - Calmodulin-dependent cyclic nucleotide phosphodiesterases in the immature rat testis. PMID- 6271750 TI - Inhibition of purified human and herpes simplex virus-induced DNA polymerases by 9-(2-hydroxyethoxymethyl)guanine triphosphate. Effects on primer-template function. AB - The inhibition of highly purified herpes simplex virus (HSV)-induced and host cell DNA polymerases by the triphosphate form of 9-(2-hydroxyethoxymethyl)guanine (acyclovir; acycloguanosine) was examined. Acyclovir triphosphate (acyclo-GTP) competitively inhibited the incorporation of dGMP into DNA, catalyzed by HSV DNA polymerase; apparent Km and Ki values of dGTP and acyclo-GTP were 0.15 microM and 0.003 microM, respectively. HeLa DNA polymerase alpha was also competitively inhibited; Km and Ki values of dGTP and acyclo-GTP were 1.2 microM and 0.18 microM, respectively. In contrast, HeLa DNA polymerase beta was insensitive to the analogue. The "limited" DNA synthesis observed when dGTP was omitted from HSV or alpha DNA polymerase reactions was inhibited by acyclo-GTP in a concentration dependent manner. Prior incubation of activated DNA, acyclo-GTP, and DNA polymerase (alpha or HSV resulted in a marked decrease in the utilization of the primer-template in subsequent DNA polymerase reactions. This decreased ability of preincubated primer-templates to support DNA synthesis was dependent on acyclo GTP, enzyme concentration, and the time of prior incubation. Acyclo-GMP terminated DNA was found to inhibit HSV DNA polymerase-catalyzed DNA synthesis. Kinetic experiments with variable concentrations of activated DNA and fixed concentrations of acyclo-GMP-terminated DNA revealed a noncompetitive inhibition of HSV-1 DNA polymerase. The apparent Km of 3'-hydroxyl termini was 1.1 X 10(-7) M, the Kii and Kis of acyclo-GMP termini in activated DNA were 8.8 X 10(-8) M and 2.1 X 10(-9) M, respectively. Finally, 14C-labeled acyclo-GMP residues incorporated into activated DNA by HSV-1 DNA polymerase could not be excised by the polymerase-associated 3',5'-exonuclease activity. PMID- 6271751 TI - Effect of self-association on activity of an ADP-ribosyltransferase from turkey erythrocytes. Conversion of inactive oligomers to active protomers by chaotropic salts. PMID- 6271752 TI - Reconstitution of the voltage-sensitive sodium channel of rat brain from solubilized components. AB - The voltage-sensitive sodium channel of rat brain synaptosomes was solubilized with sodium cholate. The solubilized sodium channel migrated on a sucrose density gradient with an apparent S20,w of approximately 12 S, retained [3H]saxitoxin ([3H]STX) binding activity that was labile at 36 degrees C but no longer bound 125I-labeled scorpion toxin (125I-ScTX). Following reconstitution into phosphatidylcholine vesicles, the channel regained 125I-ScTX binding and thermal stability of [3H]STX binding. Approximately 50% of the [3H]STX binding activity and 58% of 125I-ScTX binding activity were recovered after reconstitution. The reconstituted sodium channel bound STX and ScTX with KD values of 5 and 10 nM, respectively. Under depolarized conditions, veratridine enhanced the binding of 125I-ScTX with a K0.5 of 20 microM. These KD and K0.5 values are similar to those of the native synaptosome sodium channel. 125I-ScTX binding to the reconstituted sodium channel, as with the native channel, was voltage dependent. The KD for 125I-ScTX increased with depolarization. This voltage dependence was used to demonstrate that the reconstituted channel transports Na+. Activation of sodium channels by veratridine under conditions expected to cause hyperpolarization of the reconstituted vesicles increased 125I-ScTX binding 3-fold. This increased binding was blocked by STX with K0.5 = 5 nM. These data indicate that reconstituted sodium channels can transport Na+ and hyperpolarize the reconstituted vesicles. Thus, incorporation of solubilized synaptosomal sodium channels into phosphatidylcholine vesicles results in recovery of toxin binding and action at each of the three neurotoxin receptor sites and restoration of Na+ transport by the reconstituted channels. PMID- 6271753 TI - The role of fructose 2,6-bisphosphate in regulation of fructose-1,6 bisphosphatase. PMID- 6271754 TI - The regulatory component of adenylate cyclase. Purification and properties. AB - The regulatory component (G/F) of adenylate cyclase, which has been purified previously, contains three putative subunits with molecular weights of 52,000, 45,000, and 35,000 (Northup, J. K., Sternweis, P. C., Smigel, M. D., Schleifer, L. S., Ross, E. M., and Gilman, A. G. (1980) Proc. Natl. Acad. Sci. U. S. A. 77, 6516-6520). The published procedure has been modified to reduce the time required for preparation and to increase the yield. Application of the improved procedure allows purification of .5 to 1.0 mg of purified G/F from 1.5 kg of frozen rabbit liver. Greater than 95% of the protein observed on sodium dodecyl sulfate polyacrylamide gels is found in the three bands mentioned above. Purified G/F has the following properties: 1. Hydrodynamic measurements in cholate indicate that purified hepatic G/F has a molecular weight of about 70,000. If G/F is activated with either fluoride or GTP analogs, its apparent molecular weight is reduced to 50,000. 2. The measurement of G/F by reconstitution with the catalytic moiety of adenylate cyclase is dependent on the concentrations of both G/F and catalytic moiety. This interaction is consistent with a model derived from a simple bimolecular binding equilibrium. 3. Purified G/F can be activated by fluoride and guanine nucleotide analogs in a Mg2+-dependent reaction. The rate of activation by guanine nucleotides is markedly stimulated by high concentrations of Mg2+, indicating a site of action of divalent metallic cations on G/F. 4. The 52,000- and 45,000-dalton polypeptides can be partially resolved by heptylamine-Sepharose chromatography. G/F fractions that are enriched in the 52,000-dalton protein reconstitute hormone-stimulated adenylate cyclase activity more efficiently and are activated by GTP analogs more rapidly than are fractions that are essentially free of this polypeptide. The 35,000-dalton protein is present in all cases. PMID- 6271755 TI - Cyclic Nucleotide-independent protein kinases from rabbit reticulocytes. Identification and characterization of a protein kinase activated by proteolysis. PMID- 6271756 TI - Perturbation and structural organization of the two intrachain cAMP binding sites of cAMP-dependent protein kinase II. PMID- 6271757 TI - The effect of methylation on cytochrome c fragment complementation. AB - The heme propionates of heme peptide 1-25 from horse heart cytochrome c were methylated in acidified methanol. The methylated heme peptide combines with apocytochrome c in a single second order kinetic phase whose rate constant is within a factor of four of that measured for the unmethylated heme peptide. The dissociation constants for the two complexes are also within a factor of four. The methylated and unmethylated complexes exhibit similar fluorescence quenching, far ultraviolet dichroic spectra, and catalytic activities. The methylated complex has a Tm about 10 degrees C lower than that of the unmethylated complex. The charge transfer band of the methylated complex occurs at 720 nm instead of 695 nm as observed for the unmethylated complex. It is proposed that methylation of the outer heme propionate causes a small steric effect which alters the geometry of the methionine 80 ligation with the heme iron. PMID- 6271758 TI - Similarities and differences between the fibronectins of normal and transformed hamster cells. AB - Fibronectins from normal and virally transformed hamster cells were compared by several criteria. The fibronectin from transformed cells was similar to that from normal cells in being an intact dimeric glycoprotein with the ability to bind to gelatin, activated thiol-Sepharose, and cells. No evidence was found for proteolytic cleavages or abnormalities in disulfide bonding of transformed cell fibronectin. This fibronectin was also shown to be active in promoting cell attachment, elongation, and alignment. Therefore, the fibronectin produced by transformed cells is not defective. However, it was shown that the transformed cells were partially deficient in their capacity to bind fibronectins from either normal or transformed cells. This deficiency has implications for the significance of the loss of fibronectin on oncogenic transformation. Partial proteolysis of the fibronectins from normal and transformed cells gave rise to the same fragments. However, the glycosylated fragments from transformed cell fibronectin appeared somewhat larger than those from normal cell fibronectin. Analysis of fibronectin glycopeptides showed that transformation leads both to more branches per core and to a higher sialylation of the asparagine-linked complex carbohydrate side chains. PMID- 6271759 TI - Formation and turnover of triglyceride-rich vesicles in the chick liver cell. Effects of cAMP and carnitine on triglyceride mobilization and conversion to ketones. AB - Refractile cytoplasmic vesicles are formed in less than 10 h when chick liver cell monolayers are incubated with serum-free medium containing 0.9 mM oleate. These vesicles are identical in microscopic appearance to those formed in monolayers by de novo fatty acid synthesis (Tarlow, D. M., Watkins, P. A., Reed, R. E., Miller, R. S., Zwergel, E. E., and Lane, M. D. (1977) J. Cell Biol. 73, 332-353), but require about one-seventh the incubation time to achieve comparable size. After release from the cells by lysis in hypotonic medium, the vesicles can be isolated by flotation at 27,000 X g. Electron microscopy reveals that the isolated vesicles are rimmed by a membrane. Analysis of vesicles isolated from cells labeled with [14C]oleate or [14C]acetate showed that greater than 95% of their 14C content was in the form of triglyceride and that most cellular [14C]triglyceride was contained in the triglyceride-rich vesicles. Exposure of cells to dibytyryl-cAMP after removal of oleate from the medium caused the disappearance of triglyceride-rich vesicles within 36 h. In the absence of cyclic nucleotide, the vesicles persist. Consistent with this morphological change, dibutyryl-cAMP caused a 5.5-fold activation of the apparent rate of mobilization of cellular [14C]triglyceride from cells previously labeled with [14C]oleate. L-( -)-Carnitine alone had no effect; however, when added with dibutyryl-cAMP, cellular triglyceride mobilization was activated 7.4-fold. Although [14C]triglyceride was the principal 14C-labeled product secreted in the absence of cyclic nucleotide and comprised 90% of the total, [14C]acetoacetate and [14C] beta-hydroxybutyrate became major products when cells were treated with dibutyryl cAMP. Thus, dibytyryl-cAMP activated ketogenesis from cellular [14C]triglyceride by 200-fold and when added with L-(--)-carnitine, by 400-fold. Cells containing triglyceride-rich vesicles labeled with [2-glyceryl-3H]triglyceride were generated by incubation with medium containing [2-3H]glycerol. A comparison of the rates of loss of cellular [1-oleoyl-14C- and [2-glyceryl-3H]triglyceride revealed that substantial re-esterification, i.e. recycling, of 14C-fatty acid released by lipolysis occurred. Under conditions where recycling of 3H label ws minimal, it was determined that 15% of the cellular [2-glyceryl-3H]triglyceride was secreted "en bloc," i.e. without prior lipolysis. En bloc secretion was not affected by dibutyryl-cAMP. The rate of lipolysis of vesicle-associated [2 glyceryl-3H]triglyceride was increased 2.2-fold in the presence of dibutyryl cAmP. Chloroquine markedly inhibited the dibutyryl-cAMP-dependent lipolysis suggesting the participation of lysosomes in the mobilization of triglyceride rich vesicles. Mechanisms are presented which could account for the effects of cAMP and carnitine on the turnover of vesicle triglyceride both at the level of lipolysis and the utilization of the released fatty acids by mitochondria... PMID- 6271760 TI - The role of mRNA competition in regulating translation. III. Comparison of in vitro and in vivo results. AB - Competition of encephalomyocarditis virus, reovirus, and L-cell mRNAs for a message-discriminatory component was studied in vitro. The data were analyzed qualitatively to determine the relative initiation efficiencies among the various mRNAs. The effects of potassium chloride concentration, magnesium acetate concentration, and m7G methylation on mRNA competition in vitro were also studied. These results were correlated with translation rates in vivo for the same mRNAs, to determine if the sites of competition in vitro and in vivo are the same. It was found that under a particular set of magnesium acetate and potassium chloride concentrations, the order of mRNA initiation efficiencies was the same both in vivo and in vitro, suggesting that the same limiting message discriminatory factor is regulating initiation rates in both cases. This can only be accomplished in a competitive situation when RNA is in molar excess relative to the discriminatory component. PMID- 6271761 TI - The nucleotide sequence of rabbit embryonic globin gene beta 3. AB - The nucleotide sequence of a rabbit embryonic globin gene, beta 3, has been determined from 161 base pairs (bp) on the 5' side of the mRNA cap site to 209 base pairs beyond the 3' poly A addition site. The 5' and 3' ends of mRNA from both embryonic globin genes beta 3 and beta 4 have been determined by an S1 protection assay. Sequences that are highly conserved in the 5' flanking region of eukaryotic structural genes, AATAAAA and CCAAT, are located -25 to -31 nucleotides and -81 to -85 nucleotides, respectively, before the cap site. The CCAAT sequence is duplicated at -108 to -112 nucleotides, as it is in the human fetal gamma-globin genes. Small (124 bp) and large (817 bp) intervening sequences are located between codons 30 and 31 and between 104 and 105, respectively. The sequence AATAAA precedes the predominant poly(A) addition site by 19 nucleotides. Although rabbit globin gene beta 3 is transcribed and translated almost exclusively in embryonic erythrocytes, it shares striking homology with the human gamma-globin genes which are expressed in erythrocytes from fetal liver. The evolutionary conservation of rabbit beta 3 and human gamma correlates well with their similar chromosomal positions in the two genes families. PMID- 6271762 TI - Biochemical evidence for functional heterogeneity of cardiac sarcoplasmic reticulum vesicles. AB - Two subpopulations of cardiac sarcoplasmic reticulum vesicles were resolved functionally, based on their sensitivities to the drug ryanodine. These two subpopulations of sarcoplasmic reticulum vesicles, termed ryanodine-sensitive and ryanodine-insensitive, were separated by preloading crude cardiac microsomes with Ca2+ oxalate in the presence of ATP, followed by sucrose density gradient centrifugation. Ryanodine-insensitive vesicles accumulated most of the Ca2+ oxalate during the preload, and constituted the densest subfraction recovered from the sucrose gradient. These ryanodine-insensitive vesicles exhibited the highest density of Ca2+ pumps, and accounted for 10 to 15% of the total protein in crude cardiac microsomes. Ryanodine-insensitive vesicles continued to transport substantial amounts of Ca2+ after isolation. Ryanodine-sensitive vesicles accumulated negligible Ca2+ during the preload, and were recovered from the lower density regions of the sucrose gradient. On a milligrams of protein basis, these vesicles were present in 7-fold excess over ryanodine-insensitive vesicles. Ryanodine-sensitive vesicles transported low amounts of Ca2+ under normal incubation conditions, but 3 X 10(-4) M ryanodine strikingly increased their Ca2+ uptake 5- to 10-fold. Ca2+ uptake by ryanodine-sensitive vesicles was uniquely regulated by Ca2+ ion concentration. Elevation of the ionized Ca2+ concentration from 2 to 4 microM increased Ca2+ uptake by these vesicles greater than 5-fold, but had no effect on their Ca2+-dependent ATPase activity. These ryanodine- and Ca2+ concentration-dependent effects were apparent for only ryanodine-sensitive vesicles. Sodium dodecyl sulfate polyacrylamide gel electrophoresis revealed distinct differences in polypeptide staining between ryanodine-sensitive and ryanodine-insensitive vesicles, confirming by an independent method that the two populations of vesicles were different. These data provide the first biochemical evidence for functional and structural heterogeneity of cardiac sarcoplasmic reticulum vesicles. PMID- 6271763 TI - Evidence for two functional gal promoters in intact Escherichia coli cells. AB - We have used an S1 mapping assay to demonstrate that the mRNA transcripts of the Escherichia coli galactose operon found in intact E. coli cells with a defect in adenylate cyclase or the cyclic AMP receptor protein contain at their 5' end about five nucleotides more than the gal mRNA molecules made in wild type cells. The same difference between gal RNA synthesized in vitro in the absence of cyclic AMP or cyclic AMP receptor protein and gal RNA made in the presence of these factors is detected by this assay. Our results strongly suggest that the same two overlapping promoters, which we previously identified by in vitro transcription of gal DNA fragments, also control the expression of the galactose operon in intact cells. The intracellular levels of cyclic AMP determine which promoter is utilized. PMID- 6271764 TI - Mosaic structure and mRNA precursors of uteroglobin, a hormone-regulated mammalian gene. AB - The synthesis of uteroglobin in the rabbit uterus is induced by progesterone and is repressed by estrogen which has an over-riding effect over the inducer. The dual hormonal control system offers an excellent model for studying hormonal regulation of mammalian gene expression. Using a full-length uteroglobin cDNA clone as a specific hybridization probe, recombinant lambda phages containing the entire chromosomal uteroglobin gene have been isolated from a rabbit genomic DNA library. Electronmicroscopic analysis of hybrid molecules formed between the chromosomal uteroglobin gene and uteroglobin mRNA indicated the presence of 2 intervening sequences within this gene. The mosaic structure of the uteroglobin gene has been substantiated by detailed restriction mapping and Southern hybridization. The gene is 3.0 kilobases in length to code for a mature mRNA of 465 nucleotides. Northern hybridization of poly(A)-containing RNA from 4-day pregnant rabbit uterus with the full-length cDNA clone revealed the presence of uteroglobin mRNA precursors. The size of the largest precursor RNA species detected by the cDNA clone is the same as the entire chromosomal uteroglobin gene. The fidelity of the precursor RNAs was established by their ability to hybridize with specific intervening sequence probes. Thus the entire uteroglobin gene is expressed into primary RNA transcripts, which are subsequently processed into mature mRNA molecules by splicing. PMID- 6271765 TI - Monoclonal antibodies to the low density lipoprotein receptor as probes for study of receptor-mediated endocytosis and the genetics of familial hypercholesterolemia. AB - Monoclonal antibodies directed against the low density lipoprotein (LDL) receptor have been prepared by immunization of mice with a partially purified receptor from bovine adrenal cortex. Spleen cells from the mice were fused with the Sp2/0 Ag14 line of mouse myeloma cells. The most extensively studied monoclonal antibody, designated immunoglobulin-C7, reacts with the human and bovine LDL receptor, but not with receptors from the mouse, rat, Chinese hamster, rabbit, or dog. 125I-labeled monoclonal antibody binds to human fibroblasts in amounts that are equimolar to 125I-LDL. In fibroblasts from 6 of 8 patients with the receptor negative form of homozygous familial hypercholesterolemia, which have less than 5% of normal LdL binding, the amount of monoclonal antibody binding was also less than 5% of normal. Fibroblasts from the other two receptor-negative homozygotes bound an amount of monoclonal antibody that was much greater than expected on the basis of LDL binding, suggesting that these two patients produce a structurally altered receptor that binds the antibody, but not LDL. In normal fibroblasts, the receptor-bound monoclonal antibody was taken up and degraded at 37 degrees C at rapid rate similar to that for LDL. Fibroblasts from a patient with the internalization defective form of familial hypercholesterolemia bound the monoclonal antibody, but did not internalize or degrade it. The current data demonstrate the usefulness of monoclonal antibodies as probes for the study of the cellular and genetic factors involved in receptor-mediated endocytosis. PMID- 6271766 TI - Effects of 2,3-diphosphoglyceric acid on the human erythrocyte membrane phosphorylation system. AB - The effect of 2,3-diphosphoglycerate (2,3-P2-glycerate) on the phosphorylation of spectrin in solution by purified membrane cyclic AMP-independent protein kinase and in membrane preparations by the endogenous kinase was investigated. 2,3-P2 Glycerate inhibited spectrin phosphorylation both in solution and in the intact membrane. Kinetic analyses showed that 2,3-P2-glycerate had no effect on the Km for ATP but appeared to lower the Vmax of the reaction. When the effect of 2,3-P2 glycerate was examined in the presence of varying concentrations of spectrin, competitive inhibition kinetics were obtained. Interestingly, low concentrations of 2,3-P2-glycerate were found to effect the release of the membrane kinase from erythrocyte membranes. This release reaction may be related to the ability of 2,3 P2-glycerate to interfere with the interaction between the kinase and spectrin. The data suggest the possibility that the kinase may be bound to spectrin in the erythrocyte membrane. 2,3-P2-glycerate also caused the solubilization of 3 phosphoglyceraldehyde dehydrogenase, but not of cyclic AMP-dependent protein kinase. Taken together, our data indicate that 2,3-P2-glycerate may have a regulatory role in membrane protein phosphorylation and also may regulate the extent of association of the kinase with the membrane. PMID- 6271767 TI - Photoaffinity labeling of the beta-adrenergic receptor. AB - A new photoactive beta-adrenergic antagonist, p-azidobenzylcarazolol (pABC) has been synthesized by combining a carbazole moiety with a p-azido-benzyl substituent. The compound has been labeled with tritium to a specific activity of 26 Ci/mmol. In frog erythrocyte membranes, [3H]p-azido-benzylcarazolol binds to the beta-adrenergic receptor with the expected beta 2 specificity and with high affinity (KD congruent to 100 +/- 10 pM). Unlabeled p-azido-benzylcarazolol can irreversibly inactivate the [3H]dihydroalprenolol-binding activity of frog erythrocyte membranes in a photodependent manner which can be prevented by beta adrenergic agents. Incubation of frog erythrocyte membranes or digitonin solubilized preparations of these membranes or digitonin-solubilized preparations of these membranes which had been enriched in beta-adrenergic receptors by a Sepharose-alprenolol chromatography step led to covalent incorporation of radioactivity into a Mr = 58,000 peptide. Specific incorporation of [3H]pABC into the Mr = 58,000 peptide could be prevented by both beta-adrenergic agonists and antagonists. This peptide has previously been purified and shown to contain the beta-adrenergic receptor-binding site (Shorr, R. G. L., Lefkowitz, R. J., and Caron, M. G. (1981) J. Biol. Chem. 256, 5820-5826). Thus, photoaffinity labeling of the beta-adrenergic receptor protein directly identifies the same hormone binding subunit as has been isolated by conventional purification techniques. PMID- 6271768 TI - A precursor form of latent collagenase produced in a cell-free system with mRNA from rabbit synovial cells. AB - mRNA extracted from rabbit synovial fibroblasts which had been induced to produce large amounts of collagenase (EC 3.4.23.7) by urate crystals was translated in a cell-free wheat germ system. Collagenase was identified by immunoprecipitation using mono-specific antibody to rabbit synovial collagenase. In the absence of microsomal membranes, a single precursor with Mr = 59,000 was synthesized. This polypeptide was susceptible to proteolytic degradation. In the presence of canine pancreatic microsomes, the nascent protein was processed to a polypeptide with Mr = 57,000 (identical in mobility on sodium dodecyl sulfate-gel electrophoresis to the major latent collagenase secreted from cells) and was protected from tryptic digestion unless a detergent was used to disrupt the membranes. In addition to Mr = 57,000 material, cells secreted immunologically reactive latent collagenase with Mr = 61,000. High molecular weight collagenase was separated from Mr = 57,000 species by binding to concanavalin a-Sepharose, suggesting that this enzyme was a product of post-translational glycosylation. Both latent enzymes were activated by trypsin and human plasma kallikrein to Mr = 45,000 and 49,000. The evidence indicates that rabbit synovial fibroblast collagenase is synthesized and secreted as a single polypeptide zymogen, not as an enzyme-inhibitor complex. PMID- 6271769 TI - Stabilization by ATP and ADP of Escherichia coli dnaB protein activity. AB - The effect of adenine ribonucleotides on the stability of Escherichia coli dnaB protein in cellular crude extracts was studied. Stabilization of dnaB protein by ATP or ADP, but not by AMP, was manifested in that (i) the activity and yield of wild type dnaB protein is enhanced in the presence of ATP, (ii) the dnaB protein of E. coli dnaB mutants, such as groPB and dnaB252/ColE1::dnaC+, which is inactive in a dnaB complementation assay, can be isolated in active form in the presence of ATP or aDP, (iii) ATP or ADP protect the dnaB protein of an E. coli dnaBts mutant from inactivation at 37 degrees C, and (iv) inactive groPB and dnaBts protein can be reactivated partially by ATP. Thus, the stabilizing effect of ATP and ADP can be exploited for the isolated of otherwise inactive or labile mutant dnaB proteins. PMID- 6271770 TI - A new species of bound ubisemiquinone anion in QH2: cytochrome c oxidoreductase. AB - Using a combination of EPR and low temperature diffuse reflectance spectroscopy, a new species of semiquinone anion has been detected in QH2:cytochrome c oxidoreductase in submitochondrial particles under conditions of oxidant-induced extra reduction of cytochrome b. In contrast to the previously detected semiquinone anion, this new species is insensitive to antimycin but sensitive to treatment with 2,3-dimercaptopropanol and O2. The two species can easily be distinguished on the basis of their respective EPR properties since they differ in g-value, line width, and microwave power saturation behavior. It is concluded that the two species of semiquinone anion are bound to different domains on QH2:cytochrome c oxidoreductase. The existence of two different semiquinone anions in the enzyme strongly supports a mechanism of electron flow as proposed in the Q-cycle. PMID- 6271771 TI - Development of support matrices for affinity chromatography of thyroid hormone receptors. AB - Certain cellular responses to thyroid hormones appear to be mediated by non histone chromatin protein receptors. Purification of these proteins is important for an investigation of the detailed mechanisms of their regulatory role. In the present studies, we report the development of an affinity chromatographic system that can be used to purify thyroid hormone receptors solubilized from nuclei. Amine-substituted hormone analogs were prepared with D and L isomers of T3; these bind to the receptor. This finding supports the hypothesis that thyroid hormones fit into the receptor with the amino groups accessible from outside the binding site. Although L-triiodothyronine (LT3) (the naturally occurring isomer) binds more tightly (relative Kd = 1.0 nM) to the nuclear receptor than D triiodothyronine (DT3) (relative Kd = 2.0 nM), the amine-substituted analog of DT3 binds more tightly than the LT3 analog (DT3 analog, relative Kd = 40 nM; LT3 analog, relative Kd = 1500 nM). Agarose-based gels containing DT3 and LT3 covalently coupled by their amino groups were also prepared. Binding of receptor to these gels was biospecific in that it could be inhibited by prior incubation of the receptors with LT3. In addition, as predicted by the analog studies, the DT3 affinity gels were more effective than LT3 gels in adsorbing receptor. Elution of receptor from the LT3-derived gels was achieved in a predicted volume and concentration of counter-ligand in elution buffer. These results suggest that affinity chromatography can be applied to the purification of thyroid hormone receptors. PMID- 6271772 TI - Affinity chromatography of thyroid hormone receptors. Biospecific elution from support matrices, characterization of the partially purified receptor. PMID- 6271773 TI - Reconstituted adenylate cyclase from bovine brain. Functions of the subunits. PMID- 6271774 TI - Alloxan and H2O2 action on glucose metabolism in cultured fibroblasts. Generation of oxygen-containing free radicals as a mechanism of alloxan action. PMID- 6271775 TI - Stereochemical course of the hydrolysis of thymidine 5'-(4-nitrophenyl [17O,18O]phosphate) in H216O catalyzed by the phosphodiesterase from snake venom. AB - The phosphodiesterase from snake venom catalyzes the hydrolysis of the Rp diastereomer of thymidine 5'-(4-nitrophenyl [17O,18O]phosphate) in H216O with retention of configuration at phosphorus. This result is in agreement with those previously reported for the hydrolysis of chiral phosphorothioate substrates (Bryant, F. R., and Benkovic, S. J. (1979) Biochemistry 18, 2825-2828; Burgers, P. M. J., Eckstein, F., and Hunneman, D. H. (1979) J. Biol. Chem. 254, 7476 7478). The hydrolysis reaction catalyzed by this enzyme occurs via formation of a covalent nucleotidylated enzyme intermediate. PMID- 6271776 TI - Selective activation of rabbit ovarian protein kinase isozymes in rabbit ovarian follicles and corpora lutea. AB - The magnitude of activation of the type I and type II forms of cAMP-dependent protein kinase was investigated in estrous follicles and corpora lutea (CL) obtained from ovaries of control rabbits and rabbits injected acutely with human chorionic gonadotropin (hCG). To this end, a chromatographic technique which permitted quantitative evaluation of the in vivo activational state of the two forms of cAmP-dependent protein kinase was developed and verified. Results revealed that in follicles obtained from ovaries of untreated estrous rabbits, 15% of the soluble cAMP-dependent protein kinase, all of which exists as the type II isozyme, is activated. Intravenous administration of a single bolus of hCG promoted a concentration-dependent activation (in 10 min) of this protein kinase isozyme. In CL obtained from ovaries of control, 4-day pseudopregnant rabbits, 32% of the total soluble cAMP-dependent protein kinase exists as the type I form and 68% exists as the type II form. Both types of protein kinase are approximately 10% dissociated in CL from ovaries of untreated rabbits. Upon intravenous administration of hCG, only the type I form of cAMP-dependent protein kinase is further activated (in 10 min). Dissociation of this protein kinase is dependent upon the time and concentration of hCG. Preferential activation of the type I form of cAMP-dependent protein kinase in CL is also demonstrable in in vitro studies using exogenous cAMP. These data suggest that the physiological intracellular mediator of acute cAMP-regulated, hCG-triggered functions in rabbit ovarian follicles is the type II isozyme of cAMP-dependent protein kinase while in CL of 4-day pseudopregnant rabbits, it is the type I enzyme form. PMID- 6271777 TI - The covalent structure of pig kidney fructose-1,6-bisphosphatase. Sequence of a 63-residue cyanogen bromide peptide containing a phosphorylatable serine. AB - Native pig kidney fructose-1,6-bisphosphatase, in contrast to the rat liver enzyme, is not a substrate of cyclic AMP-dependent protein kinase. However, the pig kidney enzyme becomes a substrate when phosphorylation is performed in 1.6 M urea, after prior unfolding in 8 M urea. A cyanogen bromide fragment containing the phosphorylation site has been isolated and the amino acid sequence of this 63 residue peptide has been determined. This peptide has the following sequence: Leu Asp-Pro-Ala-Ile-Gly-Glu-Phe-Ile-Leu-Val-Asp-Arg-Asn-Val-Lys-le-Lys-Lys-Lys- Gly Ser(P)-Ile-Tyr-Ser-Ile-Asn-Glu-Gly-Tyr-Ala-Lys-Glu-Phe-Asp-Pro-Ala-Ile-Thr- Glu Tyr-Ile-Glu-Arg-Lys-Lys-Phe-Pro-Pro-Asp-Asn-Ser-Ala-Pro-Tyr-Gly-Ala-Arg-Tyr -Val Gly-Ser-Met. The amino acid sequence around the phosphorylated serine residue resembles those of other protein substrates of cyclic AMP-dependent protein kinase, but it is completely different from the phosphorylation site found in native rat liver fructose-1,6-bisphosphatase. PMID- 6271778 TI - Calcitonin induction of a persistent activated state of adenylate cyclase in human breast cancer cells (T 47D). PMID- 6271780 TI - Thyroid membrane ADP ribosyltransferase activity. Stimulation by thyrotropin and activity in functioning and nonfunctioning rat thyroid cells in culture. AB - Bovine thyroid membranes possess both ADP ribosyltransferase and NAD glycohydrolase activities with the same Km values for NAD and the same pH optima. In intact membranes, the ADP ribosyltransferase is limited in its extent by the amount of available membrane acceptor which can be ADP-ribosylated; in membranes solubilized with lithium diiodosalicylate, an artificial acceptor, L-arginine methyl ester, can be substituted to eliminate this limitation. The product of the ADP ribosyltransferase is a mono-ADP-ribosylated acceptor whether the intact or solubilized membrane provides the enzyme activity and whether membrane or exogenous acceptor, L-arginine methyl ester, is utilized. The intact membranes and the solubilized preparation also have an enzyme activity which can release AMP from the mono-ADP-ribosylated acceptor whether formed by the action of the membrane ADP ribosyltransferase or the A promoter of cholera toxin. The NAD glycohydrolase activity appears to represent the half-reaction of the ADP ribosyltransferase, i.e. an activity measurable substituting water for a membrane acceptor or L-arginine methyl ester. Membranes from functional rat thyroid cells in culture, i.e. cells chronically stimulated by thyrotropin and unresponsive to further additions of thyrotropin, have low ADP-ribosylation but high NAD glycohydrolase activities. In contrast, membranes from nonfunctional rat thyroid cells, i.e. cells unresponsive to thyrotropin, have high ADP-ribosylation and low NAD glycohydrolase activities. NAD hydrolysis by the NAD glycohydrolase activity cannot account for the low ADP-ribosylation activity in membranes from the functioning cells, and its low level of ADP-ribosylation can be eliminated by solubilizing the membranes and substituting an artificial acceptor, L-arginine methyl ester. The ADP ribosyltransferase activity of rat thyroid cell membrane preparations can be enhanced by thyrotropin in a dose-dependent manner but not by insulin, glucagon, hydrocortisone, adrenocorticotropin, or its glycoprotein hormone analog, human chorionic gonadotropin. It is thus suggested (i) that, in analogy to cholera toxin, thyrotropin-stimulated ADP-ribosylation may be important in the regulation of the adenylate cyclase response and (ii) that the level of membrane acceptor available for ADP-ribosylation may relate both to a stable "'activated" state of the adenylate cyclase system in cells chronically stimulated with thyrotropin and/or to a desensitized state with regard to a failure of more thyrotropin to elicit additional functional responses. PMID- 6271779 TI - Recovery of beta-adrenergic receptors following long term exposure of astrocytoma cells to catecholamine. Role of protein synthesis. AB - As a part of the process of agonist-induced desensitization, 1321N1 human astrocytoma cells lose up to 95% of their beta-adrenergic receptors, as detected by 125I-hydroxybenzylpindolol (125IHYP) binding, after 12-24 h of exposure to isoproterenol. In preconfluent cultures the loss of beta-receptors is completely reversible upon removal of isoproterenol, with receptor levels reaching 100% of control levels within 48-72 h. Addition of cycloheximide (5 micrograms/ml) upon removal of agonist does not prevent the recovery of receptors. After an initial 4 h lag, receptors accumulate in the presence of cycloheximide until the same receptor level is reached that was present at the onset of desensitization. Confluent cultures, which have a reduced number of receptors per cell, recover beta-receptors to only 60 to 70% of control levels following removal of isoproterenol. In addition, cycloheximide blocks the recovery of receptors in these cultures. The effects of cycloheximide on the accumulation of receptors during cell growth suggest that receptors are stable in preconfluent cultures and that turnover only occurs later when cultures are confluent. The data also indicate that long term exposure of cells to catecholamine results in a form of the beta-adrenergic receptor that is undetectable by 125IHYP binding but, nonetheless, retains its primary amino acid structure. The undetectable receptors appear to be retained until agonist is removed, whereupon they become detectable by 125IHYP binding with a t1/2 of about 36 h in the presence of cycloheximide. PMID- 6271781 TI - Fatty acyl coenzyme A-sensitive adenine nucleotide transport in a reconstituted liposome system. AB - The adenine nucleotide translocase was purified from bovine heart mitochondria and incorporated into membranes of phospholipid liposomes. The rate of transport of the adenine nucleotides was competitively inhibited by oleoyl coenzyme A with an approximate Ki of 1.0 microM. Significant inhibition was limited to those fatty acyl coenzyme A esters which are carnitine dependent for their oxidation in isolated mitochondria. Octanoyl coenzyme A was almost completely inactive as was palmitic acid and palmitoyl carnitine. By comparing the inhibitory characteristics of carboxyatractylate and bongkrekic acid with those of oleoyl CoA, it was determined that the fatty acyl-CoA esters could produce inhibition whether the carrier was inserted into the liposome in either the conventional (65%) or reverse (30%) orientation. The results demonstrate that the interaction of long chain fatty acyl-CoA esters with the ADP/ATP carrier in a purified reconstituted system mimics their effects with isolated mitochondria and inverted submitochondrial particles. In general, these findings are consistent with the role of acyl-CoA esters acting as natural ligands and biological effectors of the translocator. PMID- 6271782 TI - The binding site for ribosomal protein L11 within 23 S ribosomal RNA of Escherichia coli. AB - Ribosomal protein L11 of Escherichia coli was bound to 23 S rRNA and the resultant complex was digested with ribonuclease T1. A single RNA fragment, protected by protein L11, was isolated from such digests and was shown to rebind specifically to protein L11. The nucleotide sequence of this RNA fragment was examined by two-dimensional fingerprinting of ribonuclease digests. It proved to be 61 residues long and the constituent oligonucleotides could be fitted perfectly between residues 1052 and 1112 of the nucleotide sequence of E. coli 23 S rRNA. PMID- 6271783 TI - The appearance of voltage-sensitive Na+ channels during the in vitro differentiation of embryonic chick skeletal muscle cells. PMID- 6271784 TI - Mapping the substrate binding site of human C1r and C1s with peptide thioesters. Development of new sensitive substrates. PMID- 6271785 TI - Preparation and characterization of a probe for the cholecystokinin octapeptide receptor, N alpha (125I-desaminotyrosyl)CCK-8, and its interactions with pancreatic acini. PMID- 6271786 TI - Proteolytic conversion of beta-endorphin by brain synaptic membranes. Characterization of generated beta-endorphin fragments and proposed metabolic pathway. AB - This study concerned the fragmentation of beta-endorphin (beta-EP-(1-31) by synaptic membrane-bound peptidases. The peptides which accumulated during digestion of beta-endorphin by isolated synaptosomal plasma membrane preparations of rat brain were separated and isolated by high pressure liquid chromatography. Amino acid analysis of the peptide fractions indicated the formation of beta-EP (1-21), beta-EP-(2-21) (pH 7.4), beta-EP-(18-31), beta-EP-(1-14), and beta-EP-(1 13) (pH 5.0) in addition to previously identified gamma-endorphin (beta-EP-(1 17)), alpha-endorphin (beta-EP-(1-16), and their des-tyrosine fragments (Burbach, J. P. H., Loeber, J. G., Verhoef, J., Wiegant, V. M., De Kloet, E. R., and De Wied, D. (1980) Nature 283, 96-97). The beta-endorphin fragments obtained with crude or with purified synaptosomal plasma membranes differed only quantitatively. The peptidase which converted gamma-endorphin into beta-EP-(1 16), beta-EP-(1-15), beta-EP-(1-14), and beta-EP-(1-13), was considerably active at pH 5.0 and resembled carboxypeptidase A in degrading gamma-endorphin; the activity was reduced by the carboxypeptidase A inhibitor D-phenylalanine. The data supplement previous findings and allow routes to be delineated for the conversion of beta-endorphin by brain synaptic membranes. A pathway comprising the main events in the conversion processes is proposed and is discussed in relationship to the significance of beta-endorphin as a precursor for neuropeptides with distinct central activities. PMID- 6271787 TI - The light-activated GTP-dependent cyclic GMP phosphodiesterase complex of bovine retinal rod outer segments. Dark resolution of the catalytic and regulatory proteins. AB - A cyclic GMP phosphodiesterase associated with retinal rod outer segment (ROS) membranes is fully activated only in the presence of light and GTP. Activity can be easily depleted from the membranes by hypotonic washing, which in darkness removes two major soluble proteins. One of these has cGMP phosphodiesterase activity which is no longer activated by light or GTP. The other lacks phosphodiesterase activity but copurifies with the catalytic protein unless special measures are taken. In the present report, these 2 proteins removed from the ROS in darkness were resolved in a manner which was qualitatively and quantitatively dependent upon concentrations of 2-mercaptoethanol and EDTA. We designated the catalytic protein as P and the other protein G, because it has been reported to exhibit GTP-related activities. The unresolved P and G proteins behaved as a single complex on native gels, analytical ultracentrifugation, sucrose gradient sedimentation, and gel filtration. The P:G complex had the same Mr as purified P and more stable phosphodiesterase activity. Removal of G from P destabilized the catalytic activity and allowed aggregation of P. With loss of activity by purified P, multiple slow migrating protein bands appeared upon native gel electrophoresis. Destabilization of P could be partially prevented by addition of Mg2+ before physical separation of P from G. If 2-mercaptoethanol was removed simultaneously, dissociation of the P:G complex and destabilization of P were prevented. These findings imply that the G protein is essential to the catalytic stability of P when both are removed in darkness from the ROS membrane. PMID- 6271788 TI - Cyclic AMP regulation of lactate dehydrogenase. Quantitation of lactate dehydrogenase M-subunit messenger RNA in isoproterenol-and N6,O2'-dibutyryl cyclic AMP-stimulated rat C6 glioma cells by hybridization analysis using a cloned cDNA probe. AB - We have cloned DNA complementary to mRNA coding for rat C6 glioma cell lactate dehydrogenase M-subunit. Double-stranded DNA complementary to a portion of lactate dehydrogenase mRNA was inserted into the Pst I site of plasmid pBR322 by the dC.dG tailing technique and amplified in Escherichia coli HB101. A recombinant plasmid containing lactate dehydrogenase cDNA was identified by colony hybridization to a cDNA prepared from partially purified lactate dehydrogenase mRNA and by hybridization-selected translation. The recombinant plasmid (pRLD42) contains a 680 nucleotide insert of lactate dehydrogenase mRNA. Hybridization of nick-translation pRLD42 to glioma cell poly(A)+RNA separated on agarose gel and transferred to nitrocellulose exhibited Mr = 5.9 X 10(5) for lactate dehydrogenase mRNA. Furthermore, Northern blot analysis of RNA from unstimulated and isoproterenol-stimulated glioma cells indicated a 2-fold increase of lactate dehydrogenase mRNA molecules in stimulated cells. The 2-fold increase of lactate dehydrogenase mRNA was confirmed by RNA-excess kinetic hybridization using pRLD42 DNA and poly(A)+RNA from unstimulated, isoproterenol-, and dibutyryl cAMP-stimulated glioma cells. These data demonstrate that isoproterenol and dibutyryl cAMP cause an increase of the number of lactate dehydrogenase M-subunit mRNA molecules in glioma cells which, in part, determines the extent of synthesis of the lactate dehydrogenase M-subunit. PMID- 6271789 TI - DNA-mediated gene transfer without carrier DNA. AB - DNA-mediated gene transfer is a procedure which uses purified DNA to introduce new genetic elements into cells in culture. The standard DNA-mediated gene transfer procedure involves the use of whole cell DNA as carrier DNA for the transfer. We have modified the standard DNA-mediated gene transfer procedure to transfer the Herpes simplex virus type 1 thymidine kinase gene (TK) into TK- murine recipient cells in the absence of whole cell carrier DNA. The majority (8/10) of carrier-free transformant lines expressed the TK+ phenotype stably, in sharp contrast to our results with carrier-containing DNA-mediated gene transfer. There was a wide range in donor DNA content among independent transformants. Further analysis on one transformant line using DNA restriction digests and in situ hybridization provided evidence that, in the absence of whole cell carrier DNA, multiple donor DNA sequences became integrated at a single chromosomal site. PMID- 6271790 TI - Biochemical studies of the excitable membrane of Paramecium tetraurelia VI. Endogenous protein substrates for in vitro and in vivo phosphorylation in cilia and ciliary membranes. AB - The endogenous protein kinases of isolated Paramecium tetraurelia cilia phosphorylated approximately 30 ciliary polypeptides in vitro. Labeling with [gamma-32P]ATP was not proportional to the amount of each protein in cilia; some minor polypeptides (e.g., 67,000 and 180,000 mol wt) were more heavily labeled than some major polypeptides. Certain of the endogenous substrates for protein kinase were localized in the ciliary membrane (130,000, 86,000, 67,000, and 45,000 mol wt); others were found in axonemes or in both fractions. With cilia from bacterized cultures in the undefined Cerophyl medium, the labeling of specific endogenous phosphate acceptors was altered by pH, cyclic AMP, and cyclic GMP, but the labeling pattern was not affected by the presence of Na+ or K+ (15 mM), Ba++ (5 mM), Ca++ (10(-5) or 10(-4) M), or EGTA. Very similar results were obtained with cilia from cells grown axenically in a semidefined medium; the molecular weights and the extent of phosphorylation of the phosphopolypeptides were comparable to those of cilia from bacterized Cerophyl cultures, although no significant cyclic nucleotide effects were observed in the axenic cilia. Most of the phosphopolypeptides labeled in vitro also turned over rapidly in vitro. The phosphoprotein phosphatase responsible for turnover was partially inhibited by 5 mM NaF. The pattern of ciliary polypeptides labeled in vivo was similar to that observed in the in vitro experiments, although the relative intensities of labeling differed. Six behavioral mutants of Paramecium, known to have defects in the excitable membrane that regulates the ciliary beat, showed normal patterns of ciliary protein phosphorylation in vitro, with and without added cyclic nucleotides, at both pH 6.0 and pH 8.0. The mutants also had apparently normal phosphoprotein phosphatase. The Paranoiac A mutant, however, showed a reduction in cyclic GMP-stimulated protein kinase activity. PMID- 6271791 TI - Calcitonin stimulates plasminogen activator in porcine renal tubular cells: LLC PK1. AB - Plasminogen activators are highly selective proteases that activate the proenzyme plasminogen to the general protease, plasmin. We studied a porcine kidney cell line, originally isolated as a high producer of plasminogen activator, in which activities of cellular adenylate cyclase and cAMP-dependent protein kinase are increased in response to calcitonin. We found that salmon calcitonin, in the concentration range 0.03-300 nM, increased plasminogen activator production up to approximately 1,000-fold and concurrently inhibited cell multiplication; both of these effects were reversible. Human calcitonin was approximately 0.01 times as potent as salmon calcitonin, corresponding to potency differences observed in other biological systems. Plasminogen activator production was also stimulated by other agents that raise cellular cAMP levels such as cholera toxin, phosphodiesterase inhibitors, and vasopressin, but not to the same extent as by calcitonins. The rapidity and sensitivity of the plasminogen activator determination and other cellular responses may make it possible in the future to use this cell stain in a convenient bioassay for calcitonins and their analogues. PMID- 6271792 TI - Ultrastructural localization of cyclic GMP and cyclic AMP in rat striatum. AB - The subcellular localization of cyclic GMP and cyclic AMP in the rat caudate putamen has been studied using horseradish peroxidase immunocytochemistry. Both of the putative neurotransmitter second messengers were visualized in neurons and glial cells at light microscopic resolutions, but not all cells of either category gave detectable staining. This was confirmed at the ultrastructural level where both stained and unstained elements of the same cell type were found within the same field. A striking variation was seen in cyclic nucleotide staining intensity within individual neural and glial cells. Both of the cyclic nucleotides were detected within postsynaptic terminal boutons and within astroglial processes. Cyclic GMP postsynaptic staining was stronger than glial staining, whereas the localization pattern was reversed for cyclic AMP. The synaptic localization of cyclic AMP and cyclic GMP immunoreactivity adds support to the idea that these compounds have an influential role in synaptic function within the striatum. PMID- 6271793 TI - Potassium ion influx and Na+,K+-ATPase activity are required for the hamster sperm acrosome reaction. AB - The role of a K+ ion influx and Na+,K+-ATPase activity in the hamster sperm acrosome reaction (AR) was examined, using a range of concentrations of K+,K+ ionophores and a Na+,K+-ATPase inhibitor. Washed epididymal hamster sperm, capacitated in vitro in an artificial medium containing 2 mM Ca2+, 147 mM Na+, and 3, 6, 12, 18, or 24 mM K+, began undergoing the AR after 3 h of incubation. Sperm incubated in low K+ (0.9 mM) failed to undergo the AR even after 5 h of incubation. Sperm in 0.9 mM K+ could be induced to undergo the AR when either K+ (12 mM) alone or K+ (12 mM) with 0.1 microM nigericin was added after 3.5 h of incubation. The addition of K+ alone stimulated the AR in 30 min, whereas nigericin plus K+ stimulated the AR 15 min after addition. Neither nigericin added alone (0.9 mM K+) nor nigericin plus 12 mM K+ added to a low Ca2+ (0.35 mM) system resulted in acrosome reactions. Valinomycin (1 nM) did not stimulate the AR when added together with K+ (3-24 mM) to sperm incubated in 0.9 mM K+ for 3.5 h but markedly decreased sperm motility. Micromolar levels of ouabain blocked the AR when added between t = 0--3 h to sperm incubated with 3-24 mM K+. Inhibition of AR by the addition of 1 microM ouabain to sperm incubated with 3 mM K+ was completely reversed by the addition of 0.1 microM nigericin at t = 3.5 h. These results suggest that Na+,K+-ATPase activity and the resulting K+ influx are important for the mammalian sperm AR. Some similarities between requirements for the hamster sperm AR and secretory granule exocytosis are discussed. PMID- 6271795 TI - Multiple mechanisms of growth inhibition by cyclic AMP derivatives in rat GH1 pituitary cells: isolation of an adenylate cyclase-deficient variant. AB - GH pituitary cells have been widely utilized for studies of hormone response mechanisms. Studies reported here were motivated by the desirability of isolating characterized GH clones defective in cyclic AMP synthesis or action. Spontaneously occurring GH1 cell variants resistant to the growth-inhibitory effects of cyclic AMP analogs were isolated. Characterization of four variants showed that these were deficient in adenosine kinase and had acquired resistance to the cytotoxic effects of purine nucleoside derivatives formed in the culture medium. A second-stage selection was undertaken with mutagenized adenosine kinase deficient cells. One 8 Br cAMP-resistant variant was found to have normal cyclic AMP-dependent protein kinase activity but exhibited altered adenylate cyclase activity. Activation of cyclase activity by fluoride, guanyl nucleotides, cholera toxin, and hormone (VIP) was subnormal in the variant. Mn-dependent cyclase activity was also subnormal, suggesting that the 8 Br cAMP-resistant variant may have a deficiency in the catalytic moiety of adenylate cyclase. Surprisingly, adenosine 3':5'-monophosphate and 5'-monophosphate derivatives were found to be equally potent in growth-inhibiting adenosine kinase-deficient cells. Cross resistance to 8 Br AMP was observed in the 8 Br cAMP-resistant variant. We conclude that cyclic AMP derivatives inhibit growth of GH cells by an unanticipated mechanism that is, nonetheless, related to endogenous cyclic AMP synthesis. PMID- 6271794 TI - On the mechanism of membrane damage by Staphylococcus aureus alpha-toxin. AB - Rabbit or human erythrocytes lysed with Staphylococcus aureus alpha-toxin were solubilized with Triton X-100, and the toxin was subsequently isolated by gel chromatography, sucrose density gradient centrifugation, and reincorporation into liposomes. In the presence of Triton X-100, the toxin exhibited a sedimentation coefficient of 11S and eluted at a position between those of IgG and alpha 2 macroglobulin in gel chromatography. A single polypeptide subunit of 34,000 mol wt was found in SDS PAGE. In the electron microscope, ring-shaped or cylindrical structures were observed, 8.5-10 nm in diameter, harboring central pits or channels 2-3 nm in diameter. An amphiphilic nature of these structures was evident from their capacity to bind lipid and detergent, aggregation in the absence of detergents, and low elutability from biological and artificial membranes through ionic manipulations. In contrast to the membrane-derived form of alpha-toxin, native toxin was a water-soluble, 34,000 mol wt, 3S molecule, devoid of an annular structure. Because studies on the release of radioactive markers from resealed erythrocyte ghosts indicated the presence of circumscribed lesions of approximately 3-nm effective diameter in toxin-treated membranes, the possibility is raised that native alpha-toxin oligomerizes on and in the membrane to form an amphiphilic annular complex that, through its partial embedment within the lipid bilayer, generates a discrete transmembrane channel. PMID- 6271797 TI - Glycolipids: receptors for fibronectin? AB - We have examined the hypothesis that glycolipids might serve as receptors for the cell surface glycoprotein fibronectin using three different biological assay systems. We find that purified solubilized gangliosides inhibit fibronectin mediated hemagglutination, cell spreading, and restoration of a normal morphologic phenotype to transformed cells. The inhibition is dose-dependent and competitive; hemagglutination by 2 micrograms/ml fibronectin is half-maximally inhibited by less than 1 microM gangliosides. The most effective ganglioside inhibitors generally contain the most sialic acid residues. The isolated oligosaccharide portions of gangliosides retain this inhibitory activity and the oligosaccharides with more sialic acid are more effective inhibitors. A series of other lipids or ganglioside constituents are either less effective or without detectable activity. The more active of these lipids are the more negatively charged phospholipids such as phosphatidyl serine and phosphatidyl inositol. Our results support the hypothesis that the "receptors" for fibronectin on the cell surface either consist of or contain gangliosides or other negatively charged lipids. PMID- 6271796 TI - Growth of C6 glioma cells in serum-containing medium decreases beta-adrenergic receptor number. AB - Rat C6 glioma cells were grown in 5% fetal bovine serum-containing medium and under serum-free, defined conditions. In order to ask whether cells grown in serum-free medium are phenotypically identical to cells grown in serum, we examined effects of cell growth under both conditions on the beta-adrenergic receptor, a cell surface marker that activates adenylate cyclase and thereby regulates these cells. beta-Adrenergic receptors were qualitatively similar in cells grown in serum-containing and serum-free media, but the number of receptors was 30-50% less in cells grown with serum. This effect required several days to occur or to be reversed. The decreased number of receptors appeared to be caused by an inhibitory effect of serum on receptor number and not by a stimulatory action of the constituents of the serum-free medium. Growth medium with 5% fetal bovine serum maximally inhibited beta-adrenergic receptor numbers with 1% serum causing a half-maximal inhibition. The ability of serum to inhibit the expression of beta-adrenergic receptors could be blocked by dialyzing but not by boiling fetal bovine serum. beta-Adrenergic receptor-stimulated cyclic AMP accumulation was also decreased by growth in medium containing serum. These studies demonstrate that compared to growth of cells in serum-free medium, growth in serum-containing medium can inhibit expression of cell surface beta-adrenergic receptors. These results imply that the presence of serum in medium in which cells are grown alters properties in the plasma membrane and may alter hormonal responses in such cells. PMID- 6271798 TI - Determination of ranitidine and its metabolites in human urine by reversed-phase ion-pair high-performance liquid chromatography. AB - A method using ion-pair high-performance liquid chromatography is presented for determining ranitidine, ranitidine N-oxide, ranitidine S-oxide and desmethyl ranitidine in the urine from four volunteers, given on separate occasions an intravenous and oral dose of 100 mg ranitidine. This method has been used to study the metabolism and pharmacokinetics of ranitidine by man. It was found that the elimination half-life of ranitidine ranged from 110-246 min. The mean renal clearance of ranitidine in these four volunteers was 512 ml/min. PMID- 6271799 TI - A microassay for quantitatively detecting the Epstein-Barr virus receptor on single cells utilizing flow cytometry. AB - A quantitative microassay for detecting and analyzing the Epstein-Barr virus receptor (EBVR) utilizing fluorescein-conjugated virions is presented. The test is virus substrain-specific. Both the B95-8 and P3HR-1 strains were labelled and adsorbed to a variety of targets. Relative binding of virus was assessed by flow cytometry, the results being directly comparable with those obtained by earlier methods. Cell size and cellular DNA content were measured simultaneously with virus binding, thus enabling us to calculate EBVR density and to correlate receptor synthesis and cell cycle stage. PMID- 6271800 TI - Growth and purification of SV40 virus for biochemical studies. AB - A detailed growth and purification scheme suitable for producing relatively large quantities of fully active, pure SV40 is presented together with data on recovery and purity at each step of the procedure. The scheme was designed to prevent the initial binding of virus to cell components as well as contamination of the extracted virus by cellular DNA. PMID- 6271802 TI - Urinary adenosine 3',5'-monophosphate excretion in childhood. PMID- 6271801 TI - Genetic and hormonal characterization of cryptic 21-hydroxylase deficiency. AB - Cryptic 21-hydroxylase deficiency has been previously described in asymptomatic family members of patients with classical congenital adrenal hyperplasia (CAH). These family members were detected by high baseline 17-hydroxyprogesterone levels found in the course of family studies. The hormonal responses to ACTH of the family members with cryptic 21-hydroxylase deficiency were determined and compared to the responses of patients with CAH, patients with acquired adrenal hyperplasia, family members predicted to be heterozygous for CAH, family members predicted to be unaffected, and the general population. The ACTH-stimulated levels of 17-hydroxyprogesterone and delta 4-androstenedione in the cryptic family members were elevated above the level of the general population or family members heterozygous for classical CAH, but below that of patients with CAH. The hormonal profile of patients with cryptic 21-hydroxylase deficiency is similar to that of patients with acquired adrenal hyperplasia. The response of family members heterozygous for the cryptic gene (21-OH CRYPTIC/21-OH NORMAL) was indistinguishable from that of family members heterozygous for the classical CAH gene (21-OH CAH/21-OH NORMAL). These studies support our previous proposal that patients with cryptic 21-hydroxylase deficiency are genetic compounds, having one gene for a severe enzyme deficiency and one gene for a mild 21-hydroxylase deficiency. Thus, the 21-hydroxylase genotype in cryptic 21-hydroxylase deficiency is 21-OH CAH/21-OH CRYPTIC. PMID- 6271803 TI - Goitrous hypothyroidism due to iodide-trapping defect. AB - A 32-yr-old man with goitrous hypothyroidism due to an iodide-trapping defect is described. He was admitted because of goiter which had been increasing in size. His parents were unrelated, and no cases of goiter were found in his family. On admission, serum T3 was 39 ng/dl, serum T4 was 1.0 micro g/dl, and serum TSH was 217 micro U/ml. His 24-h thyroidal 131 I uptake was 0.05%. Antithyroid antibodies were negative. In a tracer study, the thyroidal 131 I uptakes were 6.3% at 2 h, 4.0% at 6 h, and 0.9% at 24 h after iv injection of the radioiodide. The decline in the neck counts was linear and parallel to that in the serum 131 I. The 24-h urinary excretion of 131 I was 92%. The saliva to serum and gastric juice to serum ratios of 131 I concentrations at 2 h were very low (0.95 and 0.97, respectively). After the administration of iodine (14 mg in Lugol's solution/day for 10 days), serum T3 was 228 ng/dl, serum T4 was 6.8 micro g/dl, and serum TSH was 24 micro U/ml. Some biochemical studies were carried out using the patient's thyroid tissue. In a kinetic study on iodide trapping by thyroid slices, the thyroid to medium ratio of iodide concentration in the patient's tissue was constantly about 0.1, in contrast to 1.5-4.0 in a control subject. The microsomal peroxidase activity in the patient's thyroid, assessed by iodination of bovine serum albumin, was about 3-fold that in a control subject on the basis of DNA content. Both ouabain-sensitive and -insensitive thyroidal Na+ -K+ -ATPase activities were present. These results suggest that the iodide-trapping defect in this patient was due to an impairment in the specific iodide carrier system rather than in the Na+ -K+ -ATPase itself. PMID- 6271804 TI - [On the inhibition of immediate and delayed type allergic skin reactions by the drugs elevating cellular cyclic AMP (author's transl)]. PMID- 6271805 TI - Centers for Disease Control. PMID- 6271806 TI - High skeletal muscle adenylate cyclase in malignant hyperthermia. AB - Malignant hyperthermia occurs in humans with several congenital myopathies, usually in response to general anesthesia. Commonly, individuals who develop this syndrome lack symptoms of muscle disease, and their muscle lacks specific pathological changes. A biochemical marker for this myopathy has not previously been available; we found activity of adenylate cyclase and content of cyclic AMP to be abnormally high in skeletal muscle. Secondary modification of protein phosphorylation could explain observed abnormalities of phosphorylase activation and sarcoplasmic reticulum function. PMID- 6271807 TI - Virus-induced alterations in insulin release in hamster islets of Langerhans. AB - After the inoculation of Golden Syrian hamsters with the TC-83 vaccine strain of Venezuelan encephalitis (VE) virus, a sustained diminution in glucose-stimulated insulin release and glucose intolerance of shorter duration develops. To understand better the mechanism of this defect in insulin release, we examined insulin secretion in response to several test agents in isolated perifused islets from control and 24-d post-VE virus-infected hamsters. 50 islets were used in all perifusion experiments, and data were expressed as total insulin released as well as peak response for each test agent during a 30-min perifusion period from control and VE-infected islets. After perifusion with 20 mM glucose, a 45% diminution of insulin release was noted in VE-infected islets in comparison with control islets, which in turn was similar to in vivo findings. However, following 1-mM tolbutamide stimulation, insulin release was similar in control and VE infected islets. In separate studies, 1 mM tolbutamide, 10 mM theophilline, 1 mM dibutyryl cyclic (c)AMP, and 1 mM 8-bromo-cAMP resulted in statistically similar insulin-release curves in control and VE-infected islets. Additional experiments assessing [5-3H]glucose use in control and infected islets after 20 min of perifusion with 20 mM glucose revealed virtually identical values (239 +/- 30 control; and 222 +/- 27-VE-infected islets). Morphological and morphometric evaluation of VE-infected islets (21 d following virus inoculation) showed no changes in islet volume density, beta cell density, and beta cell granulation. Thus, VE virus induces a defect in glucose-stimulated insulin release from hamster beta cells that can be corrected by cAMP analogues and does not alter islet glucose use. PMID- 6271808 TI - Two independent lipoprotein receptors on hepatic membranes of dog, swine, and man. Apo-B,E and apo-E receptors. AB - We have reported previously that canine livers possess two distinct lipoprotein receptors, an apoprotein (apo)-B,E receptor capable of binding the apo-B containing low density lipoproteins (LDL) and the apo-E-containing cholesterol induced high density lipoproteins (HDLc), and an apo-E receptor capable of binding apo-E HDLc but not LDL. Both the apo-B,E and apo-E receptors were found on the liver membranes obtained from immature growing dogs, but only the apo-E receptors were detected on th hepatic membranes of adult dogs. In this study, the expression of the apo-B,E receptors, as determined by canine LDL binding to the hepatic membranes, was found to be highly dependent on the age of the dog and decreased linearly with increasing age. Approximately 30 ng of LDL protein per milligram of membrane protein were bound via the apo-B,E receptors to the hepatic membranes of 7- to 8-wk-old immature dogs as compared with no detectable LDL binding in the hepatic membranes of adult dogs (greater than 1--1.5 yr of age). Results obtained by in vivo turnover studies of canine 125I-LDL correlated with the in vitro findings. In addition to a decrease in the expression of the hepatic apo-B,E receptors with age, these receptors were regulated, i.e., cholesterol feeding suppressed these receptors in immature dogs and prolonged fasting induced their expression in adult dogs. Previously, it was shown that the apo-B,E receptors were induced in adult livers following treatment with the hypocholesterolemic drug cholestyramine. In striking contrast, the apo-E receptors, as determined by apo-E HDLc binding, remained relatively constant for all ages of dogs studied (10--12 ng/mg). Moreover, the expression of the apo-E receptors was not strictly regulated by the metabolic perturbations that regulated the apo-B,E receptors. Similar results concerning the presence of apo B,E and apo-E receptors were obtained in swine and in man. The hepatic membranes of adult swine bound only apo-E HDLc (apo-E receptors), whereas the membranes from fetal swine livers bound both LDL and apo-E HDLc (apo B,E and apo-E receptors). Furthermore, the membranes from adult human liver revealed the presence of the apo-E receptors as evidenced by the binding of 12--14 ng of HDLc protein per milligram of membrane protein and less than 1 ng of LDL protein per milligram. The membranes from the human liver also bound human chylomicron remnants and a subfraction of human HDL containing apo-E. These data suggest the importance of the E apoprotein and the apo-E receptors in mediating lipoprotein clearance, including chylomicron remnants, by the liver of adult dogs, swine, and man. PMID- 6271809 TI - Hydrogen peroxide metabolism in human monocytes during differentiation in vitro. AB - The capacity of human blood monocytes to secrete hydrogen peroxide (H2O2) and superoxide (O2-) was measured as the cells differentiated during 4 wk of culture. Morphologic transformation of monocytes into macrophages, epithelioid cells, and multinucleated giant cells accompanied a steady increase in the content of protein per cell, from 0.77 mg/10(7) cells on days 0 to 11.77 mg/10(7) cells on days 20 to 29. In contrast, secretion of H2O2 by adherent monocytes was 859 +/- 73 nmol/60 min per mg protein (mean +/- SEM, n = 18) on day 0, rose 40% on day 3, and then fell rapidly, remaining below 6% of the initial values after day 10. The decline in capacity to secrete reactive oxygen intermediates was observed whether H2O2 or O2- were measured, whether the cells were challenged with phorbol myristate acetate or with opsonized zymosan, and whether the results were expressed per milligram cell protein or per cell. Superoxide dismutase activity tripled in adherent monocytes from day 0 to day 3, and thereafter remained elevated through at least day 16. In contrast, the activity of myeloperoxidase declined rapidly, catalase and glutathione peroxidase declined more gradually, and glutathione reductase and glutathione remained constant through the period of observation. Thus, the decline in capacity to secrete H2O2 could not be attributed to increases in cellular levels of these antioxidants. On the first day of culture, H2O2 release was enhanced up to fourfold by inclusion of sodium azide or potassium cyanide in the assay medium. This enhancement appeared to be due to inhibition of monocyte myeloperoxidase, rather than catalase. This conclusion was based on the kinetics and dose-response relationships for the effects of azide and cyanide on H2O2 release and on the activities of catalase and myeloperoxidase. Thus, the differentiation of human monocytes into macrophages in vitro is accompanied by an apparent reduction in the capacity to produce H2O2 and O2-. In this regard, the human monocyte-derived macrophage comes to resemble the resting tissue macrophage previously characterized in the mouse peritoneal cavity. PMID- 6271810 TI - Role of adherence in cytopathogenic mechanisms of Entamoeba histolytica. Study with mammalian tissue culture cells and human erythrocytes. AB - The enteric pathogen, Entamoeba histolytica, appears to cause disease by adhering to and then destroying mucosal barriers. Using an in vitro method of studying the interaction of E. histolytica with target cells (Chinese hamster ovary [CHO] and human erythrocytes [RBC]), we examined the mechanism of amebic adherence and its role in lysis of target cells. Killing and phagocytosis of target cells by amebas ceases at 4 degrees C, allowing observation of adherence. Amebas adhere to CHO cells at 4 degrees C, 78.9% formed rosettes (amebas with >/=3 adherent CHO cells each) at 2 h. At 37 degrees C, cytochalasins B and D inhibit adherence of amebas to CHO cells (P < 0.0005). Amebas adhere to and kill CHO cells in media with <0.1 muM calcium and magnesium plus 10 mM EDTA, indicating that divalent cations are not required in the medium. Adherence of amebas to human RBC was not ABO blood group specific and showed greater adherence to human than bovine or sheep RBC (P < 0.005). Neither Fc nor complement receptors were found on amebas by standard rosette studies. The amebic adherence receptor is not trypsin (0.125%) sensitive nor inhibited by trypan blue (1 mM). N-acetyl-d-galactosamine (GALNAc) inhibited the adherence of amebas to CHO cells and human RBC (0.1 g/100 ml or 4.5 mM GALNAc, P < 0.005) by binding to a receptor on the amebic surface. GALNAc abolishes amebic cytolysis of target CHO cells (determined by (111)Indium oxine release from CHO cells, P < 0.001) but not amebic phagocytosis of CHO cells. By suspending ameba-CHO cells rosettes in dextran, we found that GALNAc (1%) reversibly inhibits amebic adherence (P < 0.0005) and that cytochalasins decrease amebic killing of adherent CHO cells (P < 0.025). These findings indicate that the adherence of E. histolytica to target cells requires microfilament function, is via a specific amebic receptor that has affinity for GALNAc, and is required to lyse cells. Inhibition of the adherence of E. histolytica may alter the pathogenicity of this organism. PMID- 6271811 TI - Receptor-mediated regulation of superoxide production in human neutrophils stimulated by phorbol myristate acetate. AB - Human neutrophils contain receptors for phorbol myristate acetate (PMA), a complex lipid that induces them to generate superoxide (O (2)). Binding of PMA to these receptors displays specificity, reversibility, and high affinity. The receptor's apparent KD was approximately 0.29 nM and multiple copies (approximately 2.1 +/- 0.6 x 10(5)) were present per neutrophil. We found that the timing and magnitude of the neutrophil's respiratory burst were set independently. The onset of O (2) production occurred after a lag that was inversely proportional to the initial concentration of added PMA. The extent (rate) of O (2) production was directly proportional to the fractional occupancy of the receptor by PMA. Dual regulatory controls, such as those we noted when neutrophils were stimulated by PMA, could afford metabolic stability in the face of transient or low intensity stimuli without compromising quick and powerful responses to larger disturbances. PMID- 6271812 TI - Receptors for insulinlike growth factor I are defective in fibroblasts cultured from a patient with leprechaunism. AB - We previously have demonstrated that fibroblasts from a patient with leprechaunism exhibited markedly decreased insulin binding to insulin receptors and that the ability of insulin to stimulate glucose incorporation in the patient's cells was greatly impaired. In addition, the insulinlike growth factor, multiplication-stimulating activity (MSA), also exhibited an impaired ability to stimulate glucose incorporation in the patient's fibroblasts, although in normal fibroblasts this response appears to be mediated by an insulinlike growth factor receptor. The present study examines 125I-labeled insulinlike growth factor I (IGF-I) binding to patient's and control fibroblasts. 125I-labeled IGF-I binds to a specific IGF-I receptor in normal fibroblasts. At steady state, binding was inhibited by unlabeled IGF-I, IGF-II, MSA III-2, MSA II, insulin, and proinsulin, in order of potency, but not by high concentrations of epidermal growth factor and human growth hormone, chemically unrelated polypeptides 125I-labeled IGF-I binding to patient's cells was decreased by approximately 75%, whereas binding of epidermal growth factor to its cell surface receptors was unaffected. Computer curve-fitting of untransformed equilibrium binding data suggests that the decreased binding resulted from a decreased Ka for IGF-I. The ability of the patient's IGF-I receptor to recognize insulin also appears to be altered. Impaired IGF-I binding by the leprechaun patient's fibroblasts may contribute to the abnormal biological response to insulinlike growth factors observed in vitro and to the in utero growth retardation. PMID- 6271813 TI - Ultrasound demonstration of giant malignant breast cyst undetected by xeromammography. AB - The present case is unusual because, despite the underlying fatty breast pattern, the large size of the mass, and the presence of carcinoma, an essentially normal xeromammogram was obtained. This case demonstrates the necessity of performing a subsequent ultrasound examination on any patient in whom a palpable breast mass is not detected by xeromammography. The present case report clearly demonstrates that cyst aspiration plays a vital role in the evaluation of cystic breast masses. PMID- 6271814 TI - Physiologic observations in a controlled clinical trial of the antiemetic effectiveness of 5, 10, and 15 mg of delta 9-tetrahydrocannabinol in cancer chemotherapy. Ophthalmologic implications. AB - One hundred twenty patients about to receive their first treatment with potentially nauseant cancer chemotherapy were randomized to one of six antiemetic treatments: (1) no treatment; (2) placebo; (3) prochlorperazine (PCPZ), 10 mg; (4) delta 9-tetrahydrocannabinol (THC), 5 mg; (5) THC, 10 mg; (6) THC, 15 mg. Four doses of each medication were given orally at 4-hour intervals starting 2 hours before chemotherapy. A study nurse was responsible for both objective (nurse) and subjective (patient) symptom questionnaires. Serum levels were obtained at intervals for cross-reacting cannabinoids. Physiologic measurements including intraocular pressure (IOP), blood pressure, and pulmonary function were also recorded. In summary, the patients were remarkably free of adverse physiologic effects. All intraocular pressures before and after treatment were within the normal range, although a surprising statistically significant increase in IOP occurred in the group receiving 5 mg THC. PMID- 6271815 TI - Clinical trials with antiemetic agents in cancer patients receiving chemotherapy. AB - Vomiting accompanied by nausea is a serious acute toxicity which occurs after chemotherapy with virtually every class of cancer chemotherapeutic agents. The inability to adequately alleviate this toxicity may lead to serious complications such as general malaise, weight loss, and electrolyte imbalance. We have reviewed 34 studies in which more than 2200 cancer patients were administered 25 different antiemetics for treatment of chemotherapy-induced vomiting. All patients received a variety of cancer chemotherapeutic agents given either as single agents or in combination. The antiemetic agents included phenothiazines, antihistamines, anticholinergics, benzoquinolizines, barbiturates, butyrophenones, procainamides, cannabinoids, steroids, and benzodiazepines. It is apparent from these studies that the use of conventional antiemetic agents for treating cancer chemotherapy induced vomiting is of marginal value, and the use of investigational antiemetic agents show conflicting results as to efficacy. More quantitative measures for evaluating emesis need to be defined, and the implications that a particular antiemetic therapy may be efficacious for some but not all classes of cancer chemotherapeutic agents need to be evaluated. PMID- 6271816 TI - THC therapeutic research by independent and state-sponsored investigators: a historical review. AB - A brief history of the control and use of cannabis in the United States is presented. Essential to the discussion are the federal law: the Marihuana Tax Stamp Act of 1937; the Controlled Substances Act of 1970; and the Federal Food, Drug and Cosmetic Act of 1962. On mandate of Congress in 1968, initial studies were directed to determine effects of long-term use in man. The annual reporting of the status of "Marihuana and Health" was established. In the early 1970s, the scope of research was broadened to include evaluation of THC for use in certain medical conditions. Interest in therapeutic research may have been influenced by anecdotal reports of benefit for nausea and vomiting of cancer chemotherapy and for elevated intraocular pressure of glaucoma, by the lobbying for laws to legalize marihuana by special interest groups, and by the passage of state "Controlled Substances Therapeutic Research" acts (CSTRA). A listing of approved INDs in four therapeutic categories, a chart comparing the components of laws passed by 25 states, and a bibliography of suggested reading for further contact with the subject matter are included. PMID- 6271817 TI - Mechanism for national distribution of delta 9-tetrahydrocannabinol (NSC-134454). AB - The National Cancer Institute (NCI) is initiating a national THC distribution program by applying to the FDA for its classification as a Group C investigational agent. Since THC is also a Schedule I drug, the distribution system requires strict adherence to Drug Enforcement Agency (DEA) security and safety regulations. Contrary to the usual distribution of Group C drugs, THC will not be available directly to physicians. THC will be made available to hospital pharmacies which are: (1) an NCI recognized Cancer Center (P-30 grant supported), (2) an NCI designated New Drug Study Group, (3) a member of the Council of Teaching Hospitals. Hospital pharmacies that are located in inadequately represented geographic areas when certain criteria are met by them will also be considered. Physicians desiring to prescribe THC need not have Schedule I registration, but should (1) have experience in cancer chemotherapy, (2) have a current DEA registration number, (3) agree to abide by the Guidelines for Use of THC, and (4) be registered with a participating pharmacy. A registered physician may prescribe THC by writing a Research Order for Medication on a usual prescription blank, including, in addition to normal required information, confirmation that patient consent has been obtained and the name of the hospital at which the physician is registered to prescribe THC. PMID- 6271818 TI - The California program for the investigational use of THC and marihuana in heterogeneous populations experiencing nausea and vomiting from anticancer therapy. AB - Published studies of delta 9-tetrahydrocannabinol (THC) and marihuana in the control of nausea and vomiting in cancer patients leave many unanswered questions about dosage, time of administration, technique of use, use in pediatric and geriatric patients, and continuous use for extended periods. Four separate protocols are described which will determine optimal dosage of oral THC in adults and children receiving cyclic chemotherapy and adults receiving chronic chemotherapy or radiotherapy, and the optimal dose for smoked marihuana. Side effects and efficacy will be assessed by chemotherapeutic agent or combination, type of cancer, age, sex, etc. How well THC is tolerated in the elderly and very ill are important questions to be answered. Research goals are combined with widespread therapeutic access of cancer patients to marihuana and THC. The problems of designing and sponsoring a very large-scale Phase III collaborative study involving community practitioners are outlined. The design stresses uniformity and simplicity to obtain compliance by the more than 200 investigators. PMID- 6271819 TI - The Controlled Substances Therapeutic Research Act in the state of Washington. AB - In March 1979, the Governor of the State of Washington signed legislation establishing a THC/marihuana research program within the Washington State Board of Pharmacy. By November 1980, approximately 80 physicians had enrolled more than 400 patients on the chemotherapy protocol which randomly assigned subjects to receive either THC and prochlorperazine or THC and placebo. This report describes the process of implementing the legislation, the administrative and clinical roles of project personnel, and the design of three currently operating protocols. PMID- 6271820 TI - Clinical relevance of cannabis tolerance and dependence. AB - Psychoactive drugs are often widely used before tolerance and dependence is fully appreciated. Tolerance to cannabis-induced cardiovascular and autonomic changes, decreased intraocular pressure, sleep and sleep EEG, mood and behavioral changes is acquired and, to a great degree, lost rapidly with optimal conditions. Mechanisms appear more functional than metabolic. Acquisition rate depends on dose and dose schedule. Dependence, manifested by withdrawal symptoms after as little as 7 days of THC administration, is characterized by irritability, restlessness, insomnia, anorexia, nausea, sweating, salivation, increased body temperature, altered sleep and waking EEG, tremor, and weight loss. Mild and transient in the 120 subjects studied, the syndrome was similar to sedative drug withdrawal. Tolerance to drug side effects can be useful. Tolerance to therapeutic effects or target symptoms poses problems. Clinical significance of dependence is difficult to assess since drug-seeking behavior has many determinants. Cannabis-induced super sensitivity should be considered wherever chronic drug administration is anticipated in conditions like epilepsy, glaucoma or chronic pain. Cannabis pharmacology suggests ways of minimizing tolerance and dependence problems. PMID- 6271821 TI - Bias and the cannabis researcher. AB - This report focuses on several aspects of the "drug" cannabis in our society: the historical notion of a chemical as a moral issue (i.e., good and evil) rather than a pharmacological one; the scientist as a human being as well as a witting or unwitting influencer of social policy; the statistical design and manipulation of research consciously or unconsciously for fame and fortune (grants); the research treatment "connection" as part of our drug abuse industrial complex, a billion dollar a year industry; and the covert governmental manipulation and distortion of cannabis (and other drug) data. PMID- 6271822 TI - Do plasma concentrations of delta 9-tetrahydrocannabinol reflect the degree of intoxication? AB - Plasma concentrations of THC were measured by gas-liquid chromatography and mass spectrometry following three routes of administration and correlated with clinical effects. Plasma concentrations peaked at 3 minutes after intravenous injection and then sharply declined. The peak "high" occurred at 30 minutes while plasma concentrations were declining. This lag between plasma concentration and "high" continued during most of the span of the drug's effects. The situation was quite similar following smoking, except that peak plasma concentrations were lower. After oral administration of THC, absorption was slow, with peak concentration occurring at 1 to 2 hours. Plasma concentrations were much lower. Correlations between plasma concentrations of drug and "high" were significant but not impressive. The degree of "high" was quite variable in relation to the prevailing plasma concentration. Conjunctival injection was found so long as plasma concentration of THC could be measured. Pulse rate increases occurred at lower concentration after oral administration than after the other two routes. It is unlikely that a range of plasma concentrations can be reliably equated with impaired performance. The mode of administration will become important should THC or some homolog become a therapeutic agent. PMID- 6271823 TI - The metabolism of delta 9-tetrahydrocannabinol and related cannabinoids in man. AB - The metabolism of delta 9-tetrahydrocannabinol (THC) and related cannabinoids in man has been studied in detail utilizing intravenous, oral, and smoking routes of administration. The general pattern of metabolism was the same in all studies involving THC and related cannabinoids. Microsomal hydroxylation allylic to the delta 9-THC double bond occurs, the major product resulting in formation of an 11 CH2OH moiety; minor hydroxylation occurs on the C-8 carbon. Nonmicrosomal oxidation of the resultant 11-OH-delta 9-THC to 11-nor-delta 9-THC-9-carboxylic acid and to other more polar acids generates the major terminal metabolic products. After oral administration, approximately equal quantities of THC and its highly active 11-hydroxymetabolite were formed, whereas the latter metabolite is a minor constituent after administration by intravenous or smoking routes. Initial pharmacokinetic analyses of the data show that the mean terminal-phase (beta-phase) plasma half-life after intravenous administration of THC was about 30 hours; after oral administration, it was 23 hours. No significant statistical difference was noted between men and women as to metabolic routes or plasma terminal-phase half-lives. PMID- 6271824 TI - Current status of therapeutic opportunities based on cannabinoid research. An overview. PMID- 6271825 TI - The clinical pharmacology and dynamics of marihuana cigarette smoking. AB - We have studied the dynamics of marihuana smoking, the plasma concentration of delta 9-tetrahydrocannabinol (THC), and the pharmacologic effects produced by the sequential smoking of two 1% marihuana cigarettes at a 2-hour interval. Three males and three females, experienced marihuana smokers, participated in the study. The results indicate that each subject smoked his or her two cigarettes at a similar rate. The THC plasma concentrations produced by the smoking of the second cigarette were slightly lower than those produced by the first cigarette. The levels of the psychologic "high" caused by the two cigarettes were similar. However, the first cigarette accelerated the heart twice as much as the second cigarette. Between males and females there were marked differences in the rate at which the cigarettes were smoked. In particular, males took more puffs, took them more often, and consumed the cigarettes more rapidly than females. The plasma concentrations of THC, the self-reported psychologic effects, and the heart rate acceleration produced by the smoking of the two cigarettes were identical between the sexes. PMID- 6271826 TI - The kinetics of cannabinoid distribution and storage with special reference to the brain and testis. AB - Male and female rats were given either single or repeated intramuscular injections of 2 microCi of 14C-delta 8-tetrahydrocannabinol. They were sacrificed by groups of three at regular intervals 2, 3, 4, 6, or 24 hours after the last injection. Samples of blood lung, brain and pituitary, spleen, liver, fat, testis, and ovary were removed. Some samples were pyrolysed in an automatic oxygen train system to measure 14C-CO2, which reflects total cannabinoid concentration; other samples were kept for measurements of individual cannabinoids after extraction. After a single administration of 14C-delta 8-THC, maximal concentration of total radioactivity was reached in the brain between 2 and 4 hours amounting to 6 ng/gm tissue, or 0.06 per cent of the administered dose. After two weeks of chronic administration, concentrations of radioactivity progressively increased in liver and neutral fat, while cannabinoid levels in brain and testis remained unchanged. These data illustrate the efficiency of the blood-brain and blood-testicular barrier in limiting the access and accumulation of cannabinoids in these tissues. PMID- 6271827 TI - Cardiovascular and metabolic considerations in prolonged cannabinoid administration in man. AB - Safe therapeutic use of cannabinoids for prolonged periods of time requires an appreciation of pharmacologic actions with both acute and repetitive administration. Cardiovascular effects of acute delta 9-tetrahydrocannabinol (THC) administration included increased sympathetic and reduced parasympathetic tone, although sympathetic reflex responses were impaired. Thus, supine tachycardia and increased blood pressure with upright hypotension are observed. With repetitive dosing, there is a transition from increased to decreased sympathetic activity and from decreased to increased parasympathetic activity, and blood volume substantially increases. As a result, supine bradycardia and decreased blood pressure with tolerance to orthostatic hypotension are observed. Relevance of these observations to management of patients with THC, including considerations of potential drug interactions, is discussed. In other experiments, THC and cannabidiol were found to inhibit metabolism of other drugs (antipyrine and barbiturates) metabolized by liver mixed-function oxidase enzymes. Potential inhibition of drug metabolism must be considered in evaluating responses to chemotherapeutic agents in cancer patients receiving THC and responses to coadministered anticonvulsants in epileptic patients receiving cannabidiol. PMID- 6271828 TI - Discriminative stimulus properties of delta 9-tetrahydrocannabinol: mechanistic studies. AB - delta 9-Tetrahydrocannabinol (THC) produces a multiplicity of pharmacologic effects including analgesic, antiinflammatory, anticonvulsant, antidiarrheal, antiglaucoma, antihypertensive, and sedative effects. Efforts to elucidate the neurochemical systems mediating the THC effects have used these and related endpoints. However, animal models useful for evaluating the mechanisms by which THC produces its unique subjective effects have only recently been established. The use of drugs as discriminative stimuli provides a means for studying such mechanisms, since generalization data from this test closely correlate with subjective properties observed in clinical studies. The present study examined the ability of various drugs to mimic or block the cue produced by THC in rats. In animals trained to discriminate 3.2 mg/kg THC from vehicle, generalization occurred consistently with cannabinoids such as 11-OH-THC, HHC, and nabilone. Stereoselective generalization was also obtained with isomers of a potent analgesic, nantradol; potencies were consistent with results from other endpoints. In contrast, THC cueing was not produced by agents acting on adrenergic, cholinergic, serotonergic, GABAergic, or opiate systems. Similarly, a number of drugs previously reported to antagonize various nonunique effects of THC uniformly failed to block its subjective properties. These results indicate that the subjective properties of THC are mediated through as yet unidentified neurochemical systems. PMID- 6271829 TI - Prostaglandins and cannabis--VIII. Elevation of phospholipase A2 activity by cannabinoids in whole cells and subcellular preparations. AB - The previously reported release of arachidonic acid by THC has now been demonstrated in murine Leydig cells and WI-38 human lung fibroblasts showing the generality of the effect. The release reaction could be antagonized by phospholipase A2 inhibitors such as quinacrine and quinine, suggesting that THC can stimulate the activity of this enzyme. Further evidence for this possibility was obtained by demonstrating the release effect on a subcellular fraction which contained the phospholipase A2 activity. The stimulation of this enzyme could have profound effects on prostaglandin synthesis and/or on the integrity of various membrane structures. PMID- 6271830 TI - Cannabinoids and the cholinergic system. AB - delta 9-Tetrahydrocannabinol (THC) decreases EEG activation and causes slow waves in the cat. The EEG slow-wave activity is accompanied by a concomitant decrease in acetylcholine release from the neocortex. The findings suggest that THC depresses the brain stem activating system. Large doses of delta 8- and delta 9 THC increase brain acetylcholine levels in rodents such as the mouse and rat, but this effect is not seen with minimal doses of the cannabinoids which show behavioral effects. The most dramatic change produced by THC is that brain acetylcholine utilization is reduced primarily in the hippocampus. PMID- 6271831 TI - Levonantradol-induced inhibition of acetylcholine turnover in rat hippocampus and striatum. AB - The effects of levonantradol, a structural novel cannabinoid-related analgesic, on the turnover rate of acetylcholine (TRACh) have been studied in various regions of the rat brain. Levonantradol (0.3 mg/kg subcutaneously) decreases the tRACh in the hippocampus and striatum by 53 and 30 per cent, respectively. The extent of the reduction in TRACh is dose dependent, with a maximal decrease of 80 per cent in the hippocampus and 57 per cent in the striatum following the 3 mg/kg dose. In contrast, cortical TRACh is not affected by any dose of levonantradol. This pattern of activity on cholinergic dynamics elicited by levonantradol is qualitatively similar to that of cannabinoids but differs from that previously reported for narcotic analgesics. Moreover naltrexone (2 mg/kg intraperitoneally) does not reverse the effects of levonantradol on striatal or hippocampal TRACh. This pattern of activity on cholinergic dynamics elicited by levonantradol is consistent with a cannabinoid-like rather than an opioid-like mode of action. PMID- 6271832 TI - GABA-like actions of levonantradol. AB - The interaction of levonantradol and its pharmacologically less active (+) enantiomer with GABAergic mechanisms was studied in several in vivo systems: (1) rat cerebellar cGMP, based on the inverse relationship of GABAergic activity and cGMP levels; (2) convulsions elicited by 3-mercaptopropionic acid, an inhibitor of GABA synthesis; and (3) activated dopamine synthesis in rat striatum following blockade of dopamine receptors. Levonantradol decreased rat cerebellar cGMP content at low doses (1.0 mg/kg intraperitoneally) and antagonized elevation of cGMP levels by the GABA biosynthesis inhibitor isoniazid at even lower doses (0.32 mg/kg intraperitoneally); this activity pattern is suggestive of GABAergic activity. This conclusion is also supported by levonantradol's protection of mice against the convulsant effects of 3-mercaptopropionic acid, GABAergic agents are known to antagonize the enhanced dopamine synthesis and turnover that accompany dopamine receptor blockade by neuroleptics. Levonantradol (0.047 mg/kg intravenously) stereospecifically attenuated the elevated dopa accumulation induced by haloperidol. Levonantradol is at least 100-fold more active than THC in blocking isoniazid-induced elevation of cGMP levels in rat cerebellum or haloperidol-induced enhanced dopa accumulation in rat striatum. PMID- 6271833 TI - Selective and potent analgetics derived from cannabinoids. AB - Based on the hypothesis that analgetic activity is a dissociable feature of the cannabinoid molecule, we examined modifications of the side chain, the phenolic moiety, and, most significantly, structures that lack the benzopyran functionality present in THC and (--)-9-nor-9 beta-hydroxyhexahydrocannabinol (HHC). A new grouping, the 1-methyl-4-phenylbutyloxy C-3 side chain, elaborates a unique lipopholic region. Replacement of the phenol substituent produced several derivatives which retain analgetic activity in the codeine potency range. Introduction of a weakly basic nitrogen at C-5 and deletion of the axial methyl group in the B ring, two structural changes forbidden by traditional cannabinoid SAR, resulted in a unique family of benzoquinolines with potent analgetic activity. The prototype of this series, levonantradol, exhibits potent and stereospecific analgetic and antiemetic activity. PMID- 6271834 TI - Antiemetic activity of N-methyllevonantradol and nabilone in cisplatin-treated cats. AB - The comprehensive emetic response to xenobiotics can be quantified by monitoring the physiological forces which effectuate vomiting. A unique sequence of thoracic pressure pulses, generated by the somatic muscles of ventilation, can be measured by means of a central venous catheter. This unambiguous emetic endpoint is used to record emetic latency, repetitions and duration of action, all on an established time base and dependent of stomach content. Deslanoside (160 micrograms/kg) produced emesis in 15 cats beginning in 4.4 +/- 1.9 min (mean +/- S.D.). Subsequent emetic episodes were related in time in a log-linear but biphasic manner. Cisplatin (7.5 mg/kg) had an emetic latency of 71 +/- 18 mn in 7 cats. Additional events also had a long-linear temporal relationship for up to 400 min. This quantified emetic profile of cisplatin served as a measure for assessing antiemetic activity of N-metyllevonantradol (Pfizer), nabilone (Lilly) and prochlorperazine. The cannabinoids all evinced dose dependent antiemetic activity in terms of either complete protection, or increased latency to the first emetic episode and reduced number of episodes in those animals not completely protected. By comparison prochlorperazine afforded only minimal protection. PMID- 6271835 TI - A comparison of THC, nantradol, nabilone, and morphine in the chronic spinal dog. AB - Morphine and delta 9-tetrahydrocannabinol (THC) have been shown to have certain pharmacologic characteristics in common. Among these are antinociception, hypothermia, and the suppression of precipitated abstinence in morphine-dependent rats. In the present study the effects of morphine were compared with the effects of THC and two synthetic cannabinoids, nantradol and nabilone, in both nondependent and morphine-dependent chronic spinal dogs. Single doses of THC, nantradol, and nabilone depressed the flexor and skin twitch reflexes and had a calming effect after intravenous infusion. These effects are similar to those of morphine. Morphine, nantradol, and nabilone, but not THC, depressed rectal temperature. Unlike morphine, however, the cannabinoids produced mydriasis and an increased startle response, and these effects were not antagonized by naltrexone. THC, nantradol, and nabilone suppressed withdrawal abstinence in 40-hour and maximally abstinent morphine-dependent chronic spinal dogs. The results suggest that THC, nantradol, and nabilone share some properties with morphine since they increased the latency of the skin twitch reflex and suppressed withdrawal abstinence. It is doubtful, however, that these actions of the cannabinoids are mediated through opioid receptors since they were not antagonized by naltrexone. PMID- 6271836 TI - Behavioral effects of levonantradol and nantradol in the rhesus monkey. AB - In rhesus monkeys, acute administration of levonantradol and nantradol produced signs of CNS depression, including ataxia with body sag, pupil dilation, ptosis, dozing, and reduced responsivity to external stimuli. Neither compound suppressed the morphine withdrawal syndrome; however, both alleviated the chronic abdominal contraction associated with withdrawal. The directly observable effects of these compounds were not antagonized by naloxone. When levonantradol was administered every 6 hours, marked tolerance developed to both the effects of levonantradol and nabilone and THC. No signs of withdrawal were observed when levonantradol injections were abruptly discontinued. When substituted in lieu of codeine under an intravenous drug self-administration procedure, neither levonantradol nor nantradol maintained responding at rates higher than those maintained by their vehicle. Finally, the discriminative effects of levonantradol were not equivalent to those of the narcotics ethylketazocine or etorphine. PMID- 6271837 TI - Levonantradol: a role for central prostanoid mechanisms? AB - Although delta 9-tetrahydrocannabinol (THC) possesses many pharmacologic activities, attempts to define sites of biochemical action for the natural cannabinoids have been hampered by their low solubility, their low potency, and their relative lack of biologic selectivity. We have recently described a potent, cannabinoid-related analgetic, levonantradol, which acts stereospecifically in animals to produce analgesia qualitatively similar to morphine but at 1/9 to 1/34 the dose. While levonantradol does not act at or through the opiate receptor, the finding of one-way cross tolerance in animals suggests that morphine and levonantradol influence common nociceptive pathways. This report describes a striking structural homology between PGE1 and levonantradol as elucidated by x ray and conformational studies. This observation is consistent with the generally recognized involvement of prostaglandins in pain and emesis and may have relevance to the site of levonantradol's analgetic actions. More importantly, it provides an ongoing, heuristic basis for exploring, in depth, the role of prostaglandins in the action of levonantradol and cannabinoids. PMID- 6271838 TI - Comparative trial of the antiemetic effects of THC and haloperidol. AB - A prospective, randomized and double-blinded trial of the comparative effects of delta-9-tetrahydrocannabinol (THC) and haloperidol (H) was begun in February 1980. Patients were randomized to initially receive either THC or haloperidol with cross-over to the other agent after two courses. All patients evaluated efficacy and toxicity of each agent and those patients completing the study expressed a preference for either THC or haloperidol. All patients are receiving chemotherapeutic agents known to induce severe vomiting (cis-platinum, nitrogen mustard, or doxorubicin) or have a history or retching with chemotherapy. Fifty two patients are evaluable as of October, 1980. THC and haloperidol were equally effective in controlling nausea and vomiting as judged by number of vomiting episodes, patient evaluation of efficacy, and patient preference. About 10% of patients had complete control of vomiting and a third had less than five episodes. Patients failing one of the antiemetics had good control with the other about half the time. Toxicities from THC were less well tolerated than those from haloperidol, but most patients had no serious side effects. Nonoverlapping toxicities and efficacy raise the possibility that a combination of the agents might be worthwhile. PMID- 6271839 TI - Treatment of human spasticity with delta 9-tetrahydrocannabinol. AB - Spasticity is a common neurologic condition in patients with multiple sclerosis, stroke, cerebral palsy or an injured spinal cord. Animal studies suggest that THC has an inhibitory effect on polysynaptic reflexes. Some spastic patients claim improvement after inhaling cannabis. We tested muscle tone, reflexes, strength and performed EMGs before and after double-blinded oral administration of either 10 or 5 mg THC or placebo. The blinded examiner correctly identified the trials in which the patients received THC in seven of nine cases. For the group, 10 mg THC significantly reduced spasticity by clinical measurement (P less than 0.01). Quadriceps EMG interference pattern was reduced in those four patients with primarily extensor spasticity. THC was administered to eight other patients with spasticity and other CNS lesions. Responses varied, but benefit was seen in three of three patients with "tonic spasms." No benefit was noted in patients with cerebellar disease. PMID- 6271840 TI - Electrophysiologic properties of the cannabinoids. AB - The effects of the psychoactive cannabinoid delta 9-tetrahydrocannabinol (THC) and the nonpsychoactive cannabinoid cannabidiol (CBD) were investigated comparatively on electrically caused transcallosal cortical evoked responses, electrically induced limbic after discharges, photically evoked cortical afterdischarges, spontaneous cortical focal epileptic potentials, and spinal monosynaptic reflexes. In each system, THC produced central excitation; for example, the drug's responses ranged from enhancement of synaptic transmission to precipitation of frank convulsions. In addition to central nervous system stimulation, THC usually elicited depression; the qualitative character of the effect of the drug was dependent upon the dosage and the test system. In contrast to THC, cannabidiol generated no CNS excitation: it was either depressant or inert in these test systems. The results clearly demonstrate the complexity of the CNS properties of THC and the selectivity of the depressant properties of cannabidiol; moreover, the data illustrate the wide range of neuropharmacologic responses that potentially any cannabinoid can effect. PMID- 6271841 TI - Topical delta 9-tetrahydrocannabinol and aqueous dynamics in glaucoma. AB - Systemic delta 9-tetrahydrocannabinol (THC), administered either by smoking marihuana or as synthetic THC in soft gelatin capsules, lowers ocular tension in various glaucomas, but at the expense of significant decreases in systolic blood pressure. Topical THC in light mineral oil vehicles, though effective in laboratory animals, was not shown effective in 0.05 and 0.1% topical solutions when administered to six subjects with primary open-angle glaucoma in a randomized, balanced, double-masked protocol. Light mineral oil, which has an affinity for corneal epithelium, is an optimum vehicle for administering drugs whose mechanisms of action are systemic rather than local within the eye. Further glaucoma research should therefore proceed with marihuanas containing insignificant levels of THC (less than 0.4%) and with various local delivery systems of the ocular-active cannabinoid found in Cannabis sativa. PMID- 6271842 TI - Cannabinoids in glaucoma: a primary screening procedure. AB - A procedure was developed for screening of cannabinoids for their ability to reduce intraocular pressure (IOP) using normal rabbits. Eight animals per group were used for statistical significance of data. A negative control group was used for every screen as well as a positive control with 1.5 mg/kg delta 9-THC given intravenously (I.V.). All compounds were tested by I.V. injection and IOP measurements were taken periodically for 5 hours. Data were analyzed by a computer program which takes into account the change in IOP of the control group. Following this procedure we found that delta 8-THC, delta 9-THC, cannabinol, and nabilone were active while cannabidiol was inactive. PMID- 6271843 TI - Cannabinoids in the management of severe nausea and vomiting from cancer chemotherapy. Some additional considerations. AB - Although the cannabinoids are effective antidotes to the nausea and vomiting produced by cancer chemotherapy, they also have other properties that may be of therapeutic value. These should be explored. The possibility of interactions, both useful and harmful, between the anticancer drugs and other drugs used at the same time needs intensive study. PMID- 6271845 TI - delta 9-Tetrahydrocannabinol as an antiemetic for patients receiving cancer chemotherapy. A pilot study. AB - We conducted a pilot study to ascertain the potential toxicity and possible efficacy of delta 9-tetrahydrocannabinol (THC) at the oral dose of 5 mg/m2. Over one third of the study population, which consisted of 25 patients, reported significant dysphoric reactions. Four patients (16 per cent) elected not to take THC rather than experience loss of motivation which interfered with their professional life. Paradoxically, on eight occasions nausea seemed to worsen with THC. After the first administration of THC, 18 patients (72 per cent) described less nausea and only two individuals (8 per cent) noted complete resolution of nausea. Two patients reported worsening of their nausea. Eighteen patients noted less vomiting (69 per cent) after the first administration of THC and four patients (15 per cent) reported completed resolution of their vomiting. By the third administration of THC, one of 14 patients (7 per cent) and two of 14 (14 per cent) noted complete alleviation of nausea and vomiting, respectively. Patients who scored high on the Brief Psychiatric Rating Scale, who reported euphoria, or who had psychogenic nausea and vomiting were most likely to have a favorable antiemetic response. The results of this pilot study suggest that orally administered THC is a toxic but transiently effective antiemetic when administered at 5 mg/m2. PMID- 6271844 TI - Nabilone: an effective antiemetic in patients receiving cancer chemotherapy. AB - Eighty evaluable patients receiving chemotherapy were entered on a random prospective double-blind study to evaluate the effectiveness of nabilone, a synthetic cannabinoid, compared to prochlorperazine. Most of these patients received cisplatin, a drug that universally produces severe nausea and vomiting, as part of a combination chemotherapy regimen. The patients served as their own controls, receiving either nabilone or prochlorperazine during two consecutive treatment courses with the identical chemotherapy. Side effects consisting of hypotension and lethargy were more pronounced with nabilone. Toxicity, in general, did not preclude antiemetic treatment and in no way interfered with chemotherapy. Sixty patients (75 per cent) reported nabilone to be more effective than prochlorperazine for relief of nausea and vomiting. Of these 60 patients, 46 required further chemotherapy and continued taking nabilone as the antiemetic of choice. PMID- 6271846 TI - Antiemetic effect of delta 9-tetrahydrocannabinol in chemotherapy-associated nausea and emesis as compared to placebo and compazine. AB - Fifty-five patients harboring a variety of neoplasms and previously found to have severe nausea or emesis from antitumor drugs were given antiemetic prophylaxis in a double-blind, randomized crossover fashion. delta 9-Tetrahydrocannabinol (THC), prochlorperazine, and placebo were compared. Nausea was absent in 40 of 55 patients receiving THC, in 8 of 55 patients receiving prochlorperazine, and in 5 of 55 in the placebo group. THC appeared to be more efficacious in controlling the emesis associated with cyclophosphamide, 5-fluorouracil, and doxorubicin and less so for nitrogen mustard and the nitrosourea. THC appears to offer significant control of nausea in most patients and exceeds by far that provided by prochlorperazine (P less than 0.005). PMID- 6271847 TI - Initial observations on the effects of delta 9-tetrahydrocannabinol on the plasma pharmacokinetics of cyclophosphamide and doxorubicin. AB - We studied the effect of THC upon the pharmacokinetics of cyclophosphamide (CTX) and doxorubicin (ADR). Plasma THC was determined by RIA. Plasma concentrations of CTX and ADR were measured by GLC and fluorescence, respectively. RIA confirmed plasma levels of THC greater than 20 ng/ml for patients who received THC. CTX half-life was not significantly changed with use of THC (7.7 +/- 3.6 hours without versus 5.25 +/- 2.6 hours with THC). ADR half-life with THC was greater than without THC (175 +/- 197 hours versus 92 +/- 92 hours, respectively). Total drug exposure as determined by areas under the curves were similar (12.4 +/- 6 microM . hr without versus 13.8 +/- 4 microM . hr with THC). These preliminary data suggest that RIA is reliable for assessing THC plasma concentrations. THC induces no apparent alterations of CTX or ADR pharmacokinetics. PMID- 6271849 TI - Localization of sympathetic postganglionic neurons of physiologically identified cardiac nerves in the dog. AB - Cardiac nerves were identified physiologically and injected with horseradish peroxidase in 38 dogs. Retrogradely labeled neurons were present in the greatest number in the middle cervical ganglion, whereas fewer labeled neurons were present in the stellate ganglion. Only occasional neurons in the superior cervical ganglion were labeled, and no labelphysiologically and injected with horseradish peroxidase in 38 dogs. Retrogradely labeled neurons were present in the greatest number in the middle cervical ganglion, whereas fewer labeled neurons were present in the stellate ganglion. Only occasional neurons in the superior cervical ganglion were labeled, and no labelphysiologically and injected with horseradish peroxidase in 38 dogs. Retrogradely labeled neurons were present in the greatest number in the middle cervical ganglion, whereas fewer labeled neurons were present in the stellate ganglion. Only occasional neurons in the superior cervical ganglion were labeled, and no labeled cells were found in the T3 to T6 paravertebral ganglia or in the ganglia contralateral to the nerve injected. following injections into specific cardiac nerves, retrograde labeling was widespread within the middle cervical ganglion, and the distributions of labeled neurons from different nerves overlapped considerably. In the middle cervical ganglion there was little or no regional grouping of cells projecting to specific cardiac nerves. within the stellate ganglion, however, te cardiac sympathetic cells were clustered primarily at the cranial pole near toe origin of the ventral and dorsal ansae. Mediastinal ganglia and ganglia located in cardiac nerves were frequently as heavily labeled as the ipsilateral stellate ganglion. The occurrence of heavy labeling in mediastinal and cardiac nerve ganglia indicates that these hitherto unreported ganglia play a significant role in cardiac neural regulation. These data imply that the organization of sympathetic neurons controlling the heart is much more complex than has previously been considered. PMID- 6271848 TI - In vivo cytogenetic effects of cannabinoids. AB - The cytogenetic effects of subacute oral administration of delta-9 tetrahydrocannabinol, cannabidiol, cannabinol, and placebo were assessed in 27 subjects in a prospective double-blind study. Both conventional chromosome aberration analysis and sister chromatid exchange were considered. A comparison group of 5 nondrug users was also investigated. The aneuploidy frequency decreased significantly following delta-9-tetrahydrocannabinol administration. By contrast, the frequencies of chromosome aberrations increased slightly for all three drug groups and decreased in the placebo and comparison groups although none of the predrug versus postdrug comparisons were statistically significant. No increase in the sister chromatid exchange frequency was found in any of the groups when predrug versus postdrug comparisons were made. The present study is the fourth short term prospective investigation failing to demonstrate a statistically significant increase in chromosome damage due to cannabinoid usage. However, it must be kept in mind that to date only gross chromosomal aberrations have been identified so that the more subtle chromosomal as well as molecular genetic damage has not been ruled out. PMID- 6271850 TI - The structure, distribution, and quantitative relationships of the glia in the abdominal ganglia of the horse leech, Haemopis sanguisuga. AB - The glial cells in abdominal ganglia of the horse leech Haemopis sanguisuga were studied by electron microscopy and analysed quantitatively to evaluate the suitability of this easily obtainable carnivorous species for physiological studies. Each abdominal ganglion contains eight giant glial cells, 12,000-14,000 small glial cells, and approximately 300 neurons. The giant glial cells constituted 44.6% and the small glial cells 6.4% of the ganglion's volume. The giant glial cells contain glycogen and bundles of filaments that are chiefly located in their periphery, close to the neurons into which they send processes. The small glial cells are frequently surrounded by the giant glial cells but also occur around neuronal perikarya and axon tracts, as well as against the basal lamina and connective tissue layers. The small glial cells contain lysosomes and sometimes form a trophospongium with the neurons. A system of extracellular channels, which is continuous with the basal lamina, indents the giant glial cells and extends around parts of the neurons. The extracellular channels contain a matrix that appears very similar to the basal lamina and to the cytoplasm in the processes of the small glial cells. Some of the extracellular channels contain collagen fibrils. Hemidesmosomes join the matrix-filled extracellular channels to both the neurons and the giant glial cells. Ionic lanthanum has a free access to the neurons and glial cells via the extracellular spaces and matrix-filled channels. Areas of synaptic intermingling rarely contain glial cell processes. PMID- 6271851 TI - Primary afferent distribution pattern in the marginal zone (lamina 1) of adult monkey and cat lumbosacral spinal cord. AB - Using Golgi preparations from adult cat and monkey the present study describes the general arborization pattern of primary afferent collaterals to the marginal layer (lamina I) of the dorsal horn. The dorsal root origin of the golgi impregnated afferents was verified in the cat by matching the structural properties of these fibers with horseradish peroxidase (HRP)-stained primary afferents which were filled with HRP via anterograde transport through dorsal rootlets. The marginal afferents, in both the cat and the monkey, are relatively fine-caliber fibers which emanate from the dorsolateral fasciculus of Lissauer. Superficially they form a transverse plexus of fibers which runs across the surface of the dorsal horn and is especially dense in the lateral portion of the dorsal aspect of the marginal layer where lamina I is thickest. In deeper regions of the marginal layer, marginal afferents are longitudinally oriented and have collaterals within the outer zone of lamina II. In both superficial and deep regions of the marginal layer the afferents run parallel with large numbers of marginal cell dendrites. Most marginal afferents give rise to collaterals with widely spaced, round to oval "boutons en passant" and "boutons terminaux" which are 1.5-2.0 micrometers in diameter. Structural correlation with previous physiological and anatomical studies suggest that most of these fibers are finely myelinated, A-delta, high-threshold mechanoreceptive fibers. PMID- 6271852 TI - The retinal projection to the thalamus in the cat: a quantitative investigation and a comparison with the retinotectal pathway. AB - The projection of cat retinal ganglion cells to the thalamus was examined using the method of retrograde axonal transport of horseradish peroxidase (HRP). After the injection site was determined physiologically, HRP was applied by one of three methods: iontophoretic injection of minimal amounts, single pressure injections and multiple pressure injections. Iontophoretic injections into single laminae of the dorsal part of the lateral geniculate nucleus (LGNd) revealed that laminae A and A1 receive almost exclusively axon terminals from alpha and beta cells. Single pressure injections elucidated the retinotopic organization of the LGNd. Multiple injections lead to HRP uptake in the whole LGNd including parts of adjacent thalamic nuclei and revealed that at least 77% of all retinal ganglion cells project to the thalamus. This pathway is made up of all alpha cells, all beta cells and almost half of the gamma cells. The thalamus receives its visual input predominantly from the ipsilateral temporal and the contralateral nasal retina; some alpha cells were also labeled in the contralateral temporal retina. The shape of the decussation line was analyzed and its width was found to be proportional to the average ganglion cell spacing along the dorsoventral axis of the retina. From a comparison of the retinothalamic and retinotectal pathways, an estimate of the number of cells with bifurcating axons could be given. The axons of all alpha cells, 10% of the beta cells, and every second gamma cell bifurcate; this amounts to 30% of the retinal ganglion cells. PMID- 6271853 TI - The tectopontine projection the the rat with comments on visual pathways to the basilar pons. AB - The projection from the superior and inferior colliculi to the basilar pons in the rat was studied with the technique of orthograde transport of labeled amino acids and autoradiography. Injections restricted to the medial or lateral regions of the superior colliculus gave rise to grain labeling representing terminal fields over the ipsilateral peduncular, dorsolateral, and ventrolateral regions of the caudal basilar pons and over the dorsomedial area of the contralateral nucleus reticularis tegmenti pontis (NRTP). The pontine projection from the superior colliculus to the lateral basilar pons is topographically organized; the medial superior colliculus projects primarily to the peduncular region, whereas the lateral superior colliculus terminates chiefly in ventrolateral pontine areas. A projection from the superior colliculus to the contralateral dorsomedial pontine and medial peduncular pontine regions, a previously undescribed finding, has also been shown. Descending fibers from the inferior colliculus do not appear to terminate extensively within the basilar pons but rather course adjacent to pontine cells of the dorsolateral region in the caudal pons. Pretectal nuclei project ipsilaterally to medial and lateral nuclei in the rostral and middle basilar pons, respectively. A rostrocaudal topography exists in the tectopontine projection; the pretectum projects to rostromiddle basilar pons, the superior colliculus to more caudal pontine regions, and the inferior colliculus (although sparsely) to further caudal areas. The pontine projection pattern from the colliculi and pretectum differs from the pontine afferents from the visual cortices. The findings of this study, when compared to our results from previous investigations on the pontocerebellar projection system, suggest that the tectal inputs to certain lateral cerebellar lobules are relayed primarily through NRTP rather than the basilar pons. The collicular projection to midvermal lobules of the cerebellum appear to be mediated in part by both NRTP and lateral pontine nuclei. PMID- 6271854 TI - Magnification functions and receptive field sequences for submodality-specific bands in SI cortex of cats. AB - Electrophysiological data were collected from the forelimb region of somatosensory cortex in barbiturate-anesthetized cats using low-impedance microelectrodes in long slanting trajectories. Subsequently, the brains were fixed and stained with thionin to locate the electrode trajectories and to correlate cytoarchitecture with neural activity. Confirming earlier experiments, regions of cortex, preferentially responsive to one submodality of afferent input, were observed to stretch in mediolateral bands across SI. Separate magnification functions were calculated for each band. In the forelimb region the magnification functions for the deep and cutaneous RA bands could be approximated by linear functions while the data for the cutaneous SA band was best described by a second-order equation. At least in the forelimb region this magnification function applied only along the anteroposterior dimension of the band because the body representation is anisotropic; along the anteroposterior dimension of the band, receptive field loci may change only millimeters on the skin when the electrode moves through 1 mm of cortex, whereas when an electrode moves in a mediolateral direction they change centimeters on the forearm for an equivalent cortical distance. Each representation is separated from the others by a transition zone having a minimal width less than 200 micron. Within the transition zones, neurons often have receptive field loci which are a compromise between loci found in the adjacent representations. In these zones responses are often difficult to elicit. Electrode penetrations encountering these transition zones contain data which fulfill the receptive field and modality criteria for the boundary of a cortical column. However, within the mediolateral length of each submodality band we have found no unit with a dimension greater than 200 micron which has a boundary or transition zone. Evidence for units of smaller size cannot be obtained because of limitations in the data collection techniques. Thus, within each submodality-specific band, large portions of the somatotopic map appear to be a mediolateral continuum while relatively abrupt changes occur in receptive field locus and in submodality when the electrode passes through a transition zone between adjacent bands. PMID- 6271855 TI - Reliability of computed tomography in assessing histopathological features of malignant supratentorial gliomas. AB - A new method was developed whereby close comparisons can be made between components of a computed tomography (CT) image and neuropathological findings. This was achieved by a combination of intravital (terminal) CT, postmortem CT, and whole brain sectioning of formalin-fixed tissue. The method was applied in seven cases of malignant supratentorial astrocytic gliomas (Kernohan Grades III and IV) and in one case of thalamic ependymoma. The glioblastoma-like parts of the astrocytic gliomas were usually correctly delineated by postcontrast CT, although there were exceptions to this rule. Tumor components consisting of diffusely growing malignant astrocytoma were difficult or impossible to delineate by CT. Necrotic areas within the tumors were accurately outlined by postcontrast CT. Peritumoral edema was correctly delineated by CT, but growth of diffuse astrocytoma or the presence of astrocytic gliosis within this edematous area seems difficult or impossible to evaluate by current CT techniques. PMID- 6271856 TI - Use of intravenous liposoluble contrast material for the examination of the liver and spleen in lymphoma. AB - An experimental contrast material has been developed that, after intravenous injection, selectively opacifies the spleen and liver on computed tomography (CT). The contrast material contains iodinated ester of poppy seed oil in an emulsified form. Clinical trials were recently initiated to test the value of this contrast agent in Hodgkin's and non-Hodgkin's lymphomas. We present here two of our initial patients in whom we found lesions in the spleen or liver following the infusion of the experimental contrast material that were not visualized on the precontrast and urographic contrast material enhanced CT scans. PMID- 6271857 TI - Vitamin B12 administration for milk fat synthesis in lactating dairy cows fed a low fiber diet. AB - Thirty lactating Holstein cows were in two groups in a study of effects of vitamin B12 injections on milk fat synthesis. All cows were fed a normal fiber diet for the first 28 days after calving and then adjusted gradually to a low fiber diet over the next 28 days. After adjustment to the low fiber diet, in a single reversal trail, cows received either 150 mg of vitamin B12 in the form of hydroxocobalamin intramuscularly every 7 days for 21 days or no treatment for 21 days. Daily milk yield (kg), percent milk fat, and milk fat yield (g) for the normal fiber, low fiber adjustment, low fiber control, and low fiber plus vitamin B12 treatments were 29.6, 3.59, 1192; 31.5, 2.85, 840; 28.0, 2.58, 715; and 28.8, 2.65, 760. Injections of vitamin B12 did not correct the milk fat depression associated with the low fiber diets. In addition, there was no consistent relationship between blood B12 and milk fat production. Milk fat production was highly correlated with molar percent acetate in the rumen .63 and with blood acetate concentration .74. PMID- 6271858 TI - Beta-adrenergic receptor involvement in lipolysis of dairy cattle subcutaneous adipose tissue during dry and lactating state. AB - The influence of lactation on beta-adrenergic receptor kinetics was studied with adipocytes from eight Holstein cows during two physiological states, dry period 30 days prepartum and 30 days postpartum or early lactation. Physiological state had no effect on binding kinetics of (--)-hydrogen-3 labeled dihydroalprenolol. Affinity rate constants (8.2 versus 7.2 X 10(7) min-1 M-1) and equilibrium dissociation constants (7.1 versus 7.9 nM) for both prepartum and postpartum periods were similar. In contrast, the apparent number of beta-adrenergic receptors varied with lactational state (42,154 versus 72,264 sites/cell) for dry and lactating status as estimated in assays containing 5 nM (--)-hydrogen-3 dihydroalprenolol. Glycerol release and adipocyte concentrations of adenosine 3',5'-cyclic phosphoric acid were assayed with or without 10 microM epinephrine. Epinephrine elicited greater release of glycerol in adipocytes from lactating than dry cows (3.91 versus 2.1 mumol/10(6) cells/120 min). The concentration of adenosine-3',5'-cyclic phosphoric acid rose during the first 5 min of incubation in the presence of epinephrine and then fell to base after 10 min. Maximum concentrations at 5 min were not different in adipocytes from dry and lactating cows (250 versus 280 pmol/10(6) cells). PMID- 6271859 TI - The kinetics of mineralization of human dentin in vitro. AB - The prospect of restoration of damaged dental tissues has recently attracted great interest. An understanding of the remineralization of human dentin in vitro was attempted by using super-saturated calcium phosphate solutions at sustained supersaturation by means of a constant solution composition method. The direct growth of HAP from solutions of low supersaturation on powdered whole human dentin was confirmed. Moreover, it was found that the kinetics were similar to those of the seeded growth at synthetic HAP. A high apparent activation energy pointed to a surface-controlled mechanism. PMID- 6271860 TI - The effect of herpes virus infection on the membrane potential of parotid acinar cells. AB - Monolayers of feline parotid acinar cells were infected with feline rhinotracheitis herpes virus. Membrane potentials and input resistances were measured before and during infection. Uninfected acinar cells had an average membrane potential of -51 mV and resistances from 6 to 120 mega-ohms; cells infected for more than 18 h had an average membrane potential of -19 mV and resistances from 30 to 100 mega-ohms. The time course of the decrease in membrane potential was studied and was found to commence about ten to 14 h after infection. PMID- 6271861 TI - [Current concepts of the mechanism of bacterial movement]. PMID- 6271862 TI - The effects of histamine and prostaglandin D2 on rat mast-cell cyclic AMP and mediator release. AB - The possibility that histamine may play a functional role in modulating mast-cell secretion, as has been suggested for basophil degranulation, has both physiologic and pharmacologic implications. Therefore the capacity of histamine to influence rat peritoneal mast-cell (RPMC) cyclic AMP levels and reversed anaphylatic degranulation as reflected in the release of 3H-serotonin (5-HT) was examined. To ascertain that RPMC were functionally responsive to exogenous hormonal stimulation, assessment of prostaglandin (PG) D2 effects on cyclic AMP and 5-HT release were determined in parallel. Although PGD2 (100 microM) increased cyclic AMP and inhibited 5-HT release in the presence of 50 microM aminophylline, histamine (up to 1000 microM) was ineffective was ineffective in both. However, 1000 microM histamine in the presence of 500 microM aminophylline was capable of transiently increasing RPMC cyclic AMP (for 15 to 30 sec) and under these conditions of suppressing 5-HT release. The receptor subtype involved in the suppressive actions of histamine appeared to be of the H-1 type as reflected in the capacity of specific H-1 agonists to reproduce the inhibition of 5-HT release, whereas neither H-2 agonists nor H-2 antagonists had any influence. Thus, under conditions in which phosphodiesterase enzymatic action is impaired, histamine in extremely high concentrations is able to modulate mast-cell secretion. However, it seems very unlikely that this action of histamine has any physiologic significance. PMID- 6271863 TI - A collagenase assay using [3H-methyl]collagen. PMID- 6271864 TI - Affinity chromatography of proteolytic enzymes on silica-based biospecific sorbents. AB - New biospecific sorbents for affinity chromatography of proteolytic enzymes were prepared by the attachment of the cyclopeptide antibiotics bacitracin, bacilliquin or gramicidin S to aminosilochrom via a reaction with p-benzoquinone. The content of the cyclopeptide ligands within the sorbents varied from 2 to 46 mumol/g. The sorbents prepared by this reaction were successfully applied in the purification of the carboxylic proteinases produced by fungi, Russula decolorans (a basidiomycete) and Trichoderma lignorum, as well as crude pepsin. Serine proteinases from Thermoactinomyces vulgaris, Trichoderma koningii, Trichoderma lignorum and bacilli (subtilisins) were also submitted to chromatography on these materials. The yields of purified enzymes approached quantitative levels, sometimes being higher as a result of elimination of inhibitors. An important advantage of these sorbents is their stability against the enzymes degrading the carbohydrate matrixes of affinity sorbents synthesized on the basis of agarose, dextran or cellulose derivatives. PMID- 6271865 TI - [Prostaglandins and cancer of the breast (author's transl)]. AB - To investigate the basic characteristics of PG synthesis by human breast tumors, we have examined the relationship that could exist between the PG production and the morphological, cellular and biochemical characteristics which actually constitute the criteria of diagnosis and prognosis of human breast cancer. The results that are discussed here show that raised PG production would seem to be a marker for neoplastic cells that are capable of leading to metastatic growths in the lymph nodes. The raised production of PG can be a marker of a cell population that has the capacity to be autonomous and which possesses a high potential for metastasis. PMID- 6271866 TI - Histochemical evidence for the differential surface labeling, uptake, and intracellular transport of a colloidal gold-labeled insulin complex by normal human blood cells. AB - A colloidal gold-labeled insulin-bovine serum albumin (GIA) reagent has been developed for the ultrastructural visualization of insulin binding sites on the cell surface and for tracing the pathway of intracellular insulin translocation. When applied to normal human blood cells, it was demonstrated by both visual inspection and quantitative analysis that the extent of surface labeling, as well as the rate and degree of internalization of the insulin complex, was directly related to cell type. Further, the pathway of insulin (GIA) transport via round vesicles and by tubulo-vesicles and saccules and its subsequent fate in the hemic cells was also related to cell variety. Monocytes followed by neutrophils bound the greatest amount of labeled insulin. The majority of lymphocytes bound and internalized little GIA, however, between 5-10% of the lymphocytes were found to bind considerable quantities of GIA. Erythrocytes rarely bound the labeled insulin complex, while platelets were noted to sequester large quantities of the GIA within their extracellular canalicular system. GIA uptake by the various types of leukocytic cells appeared to occur primarily by micropinocytosis and by the direct opening of cytoplasmic tubulo-vesicles and saccules onto the cell surface in regions directly underlying surface-bound GIA. Control procedures, viz., competitive inhibition of GIA labeling using an excess of unlabeled insulin in the incubation medium, preincubation of the GIA reagent with an antibody directed toward porcine insulin, and the incorporation of 125I-insulin into the GIA reagent, indicated the specificity and selectivity of the GIA histochemical procedure for the localization of insulin binding sites. PMID- 6271867 TI - Mixed respiratory viral infections during influenza A epidemics. AB - Mixed respiratory viral infections occurring in the course of 8 influenza A epidemics in the Estonian SSR between 1969 and 1978 were investigated. A total of 1638 patients were followed up. The IF method, serological test CFR and HIR and isolation of the virus on tissue cultures and chick embryos were used. Mixed infections were found in 0-77.7% of laboratory-confirmed cases, depending on the epidemic. A combination of influenza A + parainfluenza was observed most frequently during the influenza epidemics in 1971-1977 and a combination of influenza A + influenza B during the 1977-1978 epidemic. PMID- 6271868 TI - Effect of chemical water pollution on the physiological activity of microorganisms. AB - Anionactive detergents occurring in water in concentrations up to 100 micrograms . ml-1 do not exert inhibitory effects on the present microflora, either in case of heterotrophic bacteria, indicators of fecal pollution, or pathogenic and facultative pathogenic bacteria, including viruses. On the contrary, the studied bacterial species are able to decompose surfactants and utilize them as carbon source. Though, the rate of degradation is different with respect to respective substances, as well as with regard to the species of organisms, and their physiological activity. Only at higher detergent concentrations inhibition of microbial cell growth may be observed. PMID- 6271869 TI - Parallel cross-reactivity patterns of 2 sets of antigenically distinct cytochrome c peptides: possible evidence for a presentational model of Ir gene function. AB - B10.A mice were immunized with either the carboxyl terminal peptide fragment 81 104 of pigeon cytochrome c or its acetimidyl derivative and an immune response was seen with strong preference for the immunogen. Strain distribution studies and blocking with an anti-Ia monoclonal antibody indicated that the same immune response (Ir) gene and restriction element were utilized in both responses. The specificity of the responses were evaluated by restimulating in vitro with a set of cytochrome c fragments from various species. Even though the derivatized and native fragments were poorly cross-reactive, the same phylogenetic pattern was seen when pigeon cytochrome c fragment 81-104 primed cells were tested with the set of underivatized fragments and when acetimidyl pigeon cytochrome c fragment 81-104 primed cells were tested with the same set of derivatized fragments. Primed cells from a 2nd major histocompatibility complex congenic strain of mice, B10.A(5R), displayed equivalent discrimination between derivatized and native forms but showed a markedly different phylogenetic pattern of cross-reactivity. These data indicate that the immune system recognizes 2 sites on the nominal antigen. One site, which accounts for the common hierarchy and is under Ir gene control, contains residues Gln-100, and possibly other carboxyl terminal residues. The 2nd site, which effects the distinction between native and derivatized fragments, contains at least 1 lysine other than at the carboxyl terminal. The implications of these data for theories of T cell recognition and Ir gene function are discussed. PMID- 6271870 TI - Impaired regulation of Epstein-Barr virus-induced lymphocyte proliferation in rheumatoid arthritis is due to a T cell defect. AB - The rate of outgrowth of EBV-infected B lymphocytes is regulated by normal T lymphocytes. Removal of T cells from normal whole lymphoid populations (PBM) markedly shortens the outgrowth time of the remaining B lymphocytes. There is little difference in the much more rapid outgrowth of rheumatoid PBM after the removal of T cells, which suggests that RA lymphoid cells are unable to regulate this process. To determine whether RA T cells are defective, or EBV-infected RA B cells are unresponsive to regulatory signals, EBV-induced outgrowth in autologous and allogeneic mixtures of RA and normal B and T cells was evaluated, employing morphologic criteria and 3H-thymidine incorporation. The difference in outgrowth between RA and normal PBM was reproduced by reconstitution of EBV-infected B cells with mitomycin-treated autologous T cells. In cell-mixing experiments, normal T cells appropriately regulated both normal and RA B cells similarly, whereas RA T cells were defective in regulating either B cell population. Thus, the rapid outgrowth of EBV-infected rheumatoid lymphoid cells is due to defective T cell regulation. Moreover, normal regulation does not require cell proliferation. PMID- 6271871 TI - T lymphocyte immunity to reovirus: cellular requirements for generation and role in clearance of primary infections. AB - The cellular requirements for the development of an immune response to reovirus type 1 and the role of such a response in the clearance of a primary infection with that virus were explored. An Ia-bearing antigen-presenting cell requirement is demonstrated for the in vitro generation of secondary anti-reovirus cytolytic T lymphocytes (CTL). It is then shown that mice whose spleens are depleted of Ia bearing adherent cells by exposure in vivo to ultraviolet (UV) radiation exhibit depressed priming for reovirus-specific T lymphocyte function-CTL generation, delayed-type hypersensitivity reactivity, and T cell proliferative responsiveness. These UV-irradiated mice clear primary systemic reovirus infections as readily as normal mice. Further, athymic 'nude' mice show no defect in their ability to clear a reovirus infection. The implications of these findings for our understanding of the role of virus-specific T cell function in the clearance of systemic viral infections are discussed. PMID- 6271872 TI - Clonal analysis of the BALB/c T cell proliferative response to apo beef cytochrome c. AB - Murine T cell clones that proliferated specifically in response to the protein antigen apo cytochrome c were derived and maintained in continuous culture. Two distinct clonotypes were observed with respect to the proliferative responses observed when a variety of peptides prepared from several species of cytochrome c were tested. These 2 clonotypes appeared to recognize 2 different regions in the cytochrome c molecule. Only 1 of the 2 clonotypes tested demonstrated helper cell activity for antibody formation in vitro. PMID- 6271873 TI - Ir gene control of the cytotoxic T lymphocyte response to Sendai virus: H-2k mice are low responders to Sendai. AB - H-2k mice generate a secondary in vitro cytotoxic T lymphocyte response to Sendai virus 20- to 100-fold weaker than those of other haplotypes tested (H-2b,d,q,s). This immune response defect maps to both H-2K and H-2D. H-2k x H-2d F1 mice (responder x nonresponder) only lyse targets that have the d allele at H-2K and/or H-2D. H-2k targets are equally lysable with anti-Sendai antibody. Furthermore, H-2k mice demonstrate normal antibody and T cell proliferation responses to Sendai virus. The Ir gene defect therefore appears to be limited to the generation of the cytotoxic T lymphocytes. PMID- 6271874 TI - Ig biosynthesis in a human pre-B cell line. AB - Ig synthesis was investigated in a human Epstein-barr virus transformed pre-B like lymphoblastoid line, Josh 4. Translation of total extracted cellular RNA in a wheat germ cellfree system showed that the transcripts for both micrometer (the heavy chain of membrane IgM) and micros (the heavy chain of secreted IgM) were present in approximately equal proportions. Pulse-labeling of cells with 35S methionine in culture followed by immunoprecipitation and endo-N acetylglucosaminidase digestion of the precipitates demonstrated that each of these chains were synthesized and glycosylated intracellularly. No light chain synthesis was detected in either system. In addition, both mu-chains were found to be secreted. Hybrids formed between Josh 4 and 2 light chain-producing lymphoblastoid lines, RPMI-8866P (IgG kappa) and 32a1 (IgA lambda), were found to express surface IgM. Thus the provision of light chains permitted membrane expression of chains which might otherwise be degraded. These findings were discussed in relation to normal B cell development. PMID- 6271875 TI - Abnormal lymphocyte subsets of X-linked lymphoproliferative syndrome. PMID- 6271876 TI - Studies on amoebiasis in symptomless carriers. PMID- 6271877 TI - Investigation of some cases of oriental sore in Egypt. PMID- 6271878 TI - [Bilateral chylothorax and chyloperitoneum associated with signet-ring cell carcinoma of the lung (author's transl)]. PMID- 6271879 TI - [Microvillus membrane and basolateral membrane (author's transl)]. PMID- 6271880 TI - [Transverse myelopathy in systemic lupus erythematosus --report of a case with long-term follow-up (author's transl)]. PMID- 6271881 TI - Pulmonary lesions in patients undergoing open heart surgery: approach and management. PMID- 6271883 TI - Oat-cell carcinoma of the larynx. PMID- 6271882 TI - The epithelium and chronic inflammatory cells in scleroma. (An electron microscopic study). AB - The ultrastructure of 15 cases or rhinoscleroma have been studied. The epithelium showed intercellular edema, a reaction to the polysaccharide coat of intact bacilli, passing through defects in the basal lamina and subsequently inviting polymorph migration. The transitional stages between normal and 'reactive' plasma cells to the formation of the Russell body and the Unna cell were demonstrated, thus supporting the theory of an intracellular formation of the Russell bodies. PMID- 6271884 TI - A sclerosing haemangioma in the neck. PMID- 6271885 TI - Alpha-adrenergic responses in rabbit white fat cells: the influence of obesity and food restriction. AB - The present work was carried out to separate the influence of age from that of fat cell size in rabbit white fat cells, in order to assess the importance of changing cell size to the age-related decrease of epinephrine responsiveness. Epinephrine action and adrenergic receptor site activities were explored in two main groups of rabbits. One group, 5-6 months of age, was divided into three subgroups: the control, group I, was fed usual laboratory chow; group II was subjected to dietary restriction (3 months); group III was fed the usual diet with chronic administration of insulin (1.5 UI/kg per day for 3 months) known to induce fat cell size increment. The other main group, composed of 15- to 16-month old rabbits, was also divided into three subgroups: group IV, control; group V, subjected to dietary restriction (1 and one-half-2 months) after previous development of adipose mass; and group VI, dietrestricted for 4 months. The loss of epinephrine-responsiveness of rabbit fat cells can be either prevented by restricting food intake (groups III, V) or promoted by chronic insulin administration (group II). Isoproterenolinduced lipolysis was maintained whatever the fat-cell size, while the changes in the ability of epinephrine to promote lipolysis were linked to a variable alpha-adrenergic activity (increase of epinephrine-induced lipolysis promoted by the alpha-antagonist drug, phentolamine). Alpha-adrenergic responsiveness is increased in large fat cells (groups III and IV) while a reduced alpha-adrenergic activity is observed in small fat cells of underfed rabbits (group II). After dietary restriction, large fat cells (with an increased alpha-adrenergic responsiveness) were reduced in size and a significant restoration of the lipolytic effect of epinephrine was shown (group V). In conclusion, these results indicate that cell size, in addition to age, is an important factor affecting epinephrine-responsiveness in rabbit adipocytes. The loss or the recovery of the lipolytic effect of epinephrine could be explained by a modification of the alpha-receptor activity; the beta-receptor activity was less modified. PMID- 6271887 TI - Intra-articular radioactive colloidal gold (198 Au) in the treatment of rheumatoid arthritis. PMID- 6271886 TI - Factors regulating the activities of the low density lipoprotein receptor and the scavenger receptor on human monocyte-macrophages. AB - Improved techniques of cell isolation resulted in 90 to 100 million monocytes from a single donor. Addition of low density lipoprotein (LDL) to to cultures of these cells resulted in the down regulation of LDL receptor activity. Addition of malondialdehyde-altered LDL. which enters the cell through a receptor for negatively charged proteins (the scavenger receptor), produced an even greater down regulation of the LDL receptor, indicating that both receptors are present on the same cell. Within hours of adherence of the cells, there was a dramatic decrease in the activity of both receptors. LDL receptor activity was highest during the first week in culture and then declined, despite the maintenance of a constant LDL concentration in the medium. Scavenger receptor activity surpassed LDL receptor activity by the 6th day and was maximally expressed during the second week. Increasing cell density resulted in a slight increase in the activity of the LDL receptor and a dramatic increase in scavenger receptor activity. Insulin had no significant effect on either receptor. Removing serum from the culture medium for 48 hr resulted in a 3.5-fold increase in LDL receptor activity and a 2-fold decrease in scavenger receptor activity. Twenty-four hr after the cells were re-exposed to serum, the activities of both receptors essentially returned to base line. Heat-inactivation of serum was associated with an increased cholesteryl ester content of the cells and depressed receptor activities. Scavenger receptor activity appears related to the maturation of monocytes into macrophages and is promoted by increasing cell density and serum factors that are heat labile. PMID- 6271888 TI - A rare case of pericardial malignant fibrous histiocytoma. PMID- 6271889 TI - SEM observation on the basal layer of the rat seminiferous epithelium exposed with collagenase and trypsin. PMID- 6271890 TI - Lack of effect of angiotensin on levels of cyclic AMP in isolated adrenal zona glomerulosa cells from the rat. AB - The effects of pure [Asp1,Val5]- and [Asn1,Val5]-angiotensin II and also [des Asp1,Ile5]-angiotensin II (angiotensin III) on cyclic AMP and steroid outputs by dispersed rat capsular cells, comprising 95% zona glomerulosa and 5% zona fasciculata cells, have been studied. The results showed that [Asp1, Val5]-and [Asn1, VAl5]-angiotensin II, at doses between 2.5 X 10(-11) and 2 X 10(-4) mol/l, which produced typical increases in steroidogenesis, failed to increase output of cyclic AMP. This lack of effect was observed whether the nucleotide was measured by radioimmunoassay or by adrenal binding protein and under the same conditions in which 8.4 mM-K+ consistently increased the output of cyclic AMP. Instead the results showed a small but significant decrease in cyclic AMP output with angiotensin II. Similar results were obtained with incubations for 60 rather than 120 min and with medium containing a concentration of 5 or 40 g bovine serum albumin/l. Although the levels of cyclic AMP were generally higher in the presence of the phosphodiesterase inhibitor, 3-isobutyl-1-methylxanthine, the same decrease relative to basal outputs was observed with angiotensin II which increased steroidogenesis. Angiotensin III also failed to increase output of cyclic AMP at doses (2.5 X 10(-9) to 2.5 X 10(-6) mol/l) which produced increases in steroid output equivalent to those with angiotensin II. These results indicate that angiotensin II and III can act through a cyclic AMP-independent mechanism. PMID- 6271891 TI - Receptor heterogeneity in the human thyroid: differences between thyrotrophin binding sites in membrane and nuclear fractions. AB - Binding of 125I-labelled bovine TSH to crude membrane fractions of human thyroid tissue was a saturable, hormonally specific process which yielded non-linear Scatchard plots with limiting affinities of approximately 10(9) and 10(7) l/mol. Binding activity in membranes was soluble in Triton X-100, was inhibited specifically by immunoglobulins from patients with Graves's disease, and was increased by the beta-blocking drug, propranolol. In contrast, purified nuclear preparations showed a predominance of lower affinity binding, and their binding activity was insoluble in Triton and insensitive to immunoglobulins from patients with Grave's disease and to propranolol. Tryptic digestion liberated only low affinity binding activity from membranes or nuclei. It was concluded that human thyroid tissue contains independent classes of TSH-binding sites, which differ in their chemical, immunological and hormone-binding properties. PMID- 6271892 TI - Effects of bromocriptine and occlusion of nipples on prolactin receptor and lactose synthetase activity in the mammary gland of the lactating rat. AB - The response of prolactin receptor and lactose synthetase to suppression of plasma concentrations of prolactin was examined in normal and occluded (teat sealed) mammary glands of Sprague-Dawley rats, Rats, with mammary glands unilaterally occluded, were given bromocriptine (2.5 mg/kg per 12 h) between 5 and 8 post partum. Bromocriptine reduced plasma prolactin concentrations from 460.4 +/- 120.8 (mean +/- s.e.m.) to 2.56 +/- 0.89 ng/ml within 12h whilst concentrations in control rats were 553.4 +/- 110.25 ng/ml. Lactose synthetase activity declined rapidly, within 24h, in occluded glands of both groups but was maintained for 24 h in normal glands of bromocriptine-treated rats and decreased thereafter. Prolactin receptors also declined significantly within 24h in occluded glands. Desaturation of the prolactin receptor by bromocriptine treatment in vivo was compared with desaturation by exposure of membranes to MgCl2 in vitro. Both treatments enhanced prolactin binding but the increase after treatment with MgCl2 may have been partly artefactual since there was a selective loss of protein from the membranes. These results indicate that the prolactin receptor in rat mammary gland may be maintained after acute suppression of prolactin secretion. PMID- 6271893 TI - Studies on the mechanism of secretion of corticosteroids by the isolated perfused adrenal of the rat. AB - A technique for the perfusion of the rat adrenal cortex is described. With tissue culture Medium 199 the preparation was responsive in terms of steroid production of both ACTH and K+ ions. Production of corticosterone and 18 hydroxydeoxycorticosterone (18-hydroxy-DOC) was stimulated by ACTH when it was administered at rates between 5 uu.'/min and 5 mu./min. Increasing the K+ ion concentration of the perfusate from 3.6 to 5.4 and 8.9 mmol/l stimulated the production of aldosterone, 18-hydroxycorticosterone and deoxycorticosterone, although not of corticosterone or 18-hydroxy-DOC. This preparation has been used to study further the mechanism of secretion of corticosterone and 18-hydroxy-DOC. Thus, production of these two steroids was measured at different perfusion flows, varying between 0.1 and 0.6ml/min, with different levels of ACTH stimulation. Corticosterone production was significantly (P less than 0.001) increased by increasing flows both under control conditions and with ACTH was administered at constant rates of 50 uu./min or 1 mu./min. Production of 18-hydroxy-DOC was not affected by flow either under control conditions or with 50 uu. ACTH/min. However, when ACTH was administered at 1 mu./min. 18-hydroxy-DOC production was also significantly (P less than 0.001) increased by flow. The results are consistent with those obtained in previous in-vitro studies and have been interpreted as suggesting that the main mechanism of corticosterone secretion is simple diffusion. In contrast, 18-hydroxy-DOC secretion, at least at sub-maximal levels of stimulation, appears to require a more complex process. PMID- 6271895 TI - Peripheral neuropathy caused by chronic polychlorinated biphenyls poisoning. PMID- 6271894 TI - Cytomegalovirus causes a latent infection in undifferentiated cells and is activated by induction of cell differentiation. AB - Murine cytomegalovirus (MCMV) does not productively infect OTT6050AF1 BrdU, F9, or PCC4 undifferentiated murine teratocarcinoma cell lines, as shown by immunofluorescence assays for viral antigens and by plaque assays for infectious virus. However, these cells were infected by a variety of other viruses. MCMV does productively infect PYS2 and OTT F12 differentiated murine teratocarcinoma cell lines. The replication of MCMV in the pluripotent PCC4 cell line was examined in detail. Undifferentiated PCC4 cells could be differentiated when propagated in the presence of dimethylacetamide, as judged by changes in the expression of H-2 antigens on the cell surface. Several viruses, including lymphocytic choriomeningitis virus, herpes simplex virus type 1, and vesicular stomatitis virus, replicated to a similar extent in differentiated and undifferentiated PCC4 cells. MCMV did productively infect differentiated PCC4 cells. In contrast, MCMV did not produce infectious virus, viral antigens, or substantial viral RNA in undifferentiated PCC4 cells. The molecular block of MCMV replication occurred at the level of MCMV RNA transcription. Undifferentiated PCC4 cells have receptors for MCMV and bind similar amounts of radiolabeled virus as differentiated PCC4 cells. After MCMV binds to its receptors on undifferentiated cells, MCMV penetrates the plasma membrane and is transported to the cells' nuclei. MCMV DNA was present in the cytoplasm, and small amounts of MCMV RNA (less than 17 percent of that found in MCMV-infected differentiated PCC4 cells) were found in the nucleus. However, MCMV RNA was not detected in the cytoplasm of undifferentiated cells. A latent infection was established by infecting undifferentiated PCC4 cells with MCMV, inactivating residual infectivity with antibodies to MCMV, and propagating cells under conditions that maintained the undifferentiated state. These MCMV-infected undifferentiated cells did not produce infectious virus, viral antigens, or viral RNA but did contain viral DNA detectable by DNA-DNA hybridization kinetics. Latency was terminated and infectious virus was made when such undifferentiated cells were induced to differentiate. PMID- 6271896 TI - [Case report of cylindroma of the base of the tongue]. PMID- 6271897 TI - Structural analysis of animal virus genomes, The fifth Fleming lecture. PMID- 6271898 TI - Studies on the restriction of ecotropic murine retrovirus replication in mouse teratocarcinoma cells. AB - Retrovirus infection of cultured murine teratocarcinoma cells depends upon the state of differentiation. We have used two cell lines derived from a teratocarcinoma of mouse, strain 129. One, an undifferentiated pluripotential cell line (PCC4), is restrictive to viral infection, while the other, a differentiated myoblast-derived cell line (PCD1), is fully permissive to virus replication. We have shown that no virus RNA expression can be found in PCC4 cells 48 h post-infection and that no nucleic acid sequences can be found in an integrated form in PCC4 cells. However, the kinetics of formation of free proviral intermediates show that the three forms (I, II and III) of free virus DNA are synthesized in both PCC4 and PCD1. Free proviral DNA disappears gradually after 24 h in PCC4 cells while all forms increase in PCD1. These results suggest that the viral multiplication restriction occurs somewhere between the proviral DNA synthesis and integration of DNA in the cellular genome. PMID- 6271899 TI - The detection of DNA tumour virus-specific RNA sequences in abnormal human cervical biopsies by in situ hybridization. AB - We have used the technique of in situ nucleic acid hybridization and autoradiography of thin frozen sections of human tissue to search for virus RNA sequences in human cervical tumours. Of cervical biopsies with abnormal cytology, 67% bound herpes simplex virus type 2 (HSV2) 3H-labelled DNA probes and 39% bound adenovirus type 2 (Ad2) 3H-labelled DNA probes, whereas control experiments with phage lambda 3H-labelled DNA probes, under the same conditions, bound to only 7% of cases. In contrast, normal cervical biopsies bound the three probes in only 23%, 17% and 8% of cases respectively. PMID- 6271900 TI - Polypeptides of infectious bronchitis virus. I. Polypeptides of the virion. AB - Infectious bronchitis virus (IBV), strain Beaudette, grown in cultured cells contained five structural proteins with apparent mol. wt. of 170 000 (p170), 94 000 (gp94), 50 000 (pp50) 30 000 (gp30) and 26 000 (p26). Both gp94 and gp30 are glycopeptides since they were labelled with [3H]glucosamine. The only phosphorylated polypeptide was pp50, and both it and gp94 were occasionally resolved into two bands. Two other polypeptides with mol. wt. of 28 000 (p28) and 14 000 (p14) were sometimes associated with the virus. In egg-grown virus two additional proteins were found with mol. wt. of 110 000 (p110) and 75 000 (gp75). The cell protein, actin, was also found in highly purified IBV virions. Different serotypes of either tissue culture-grown or egg-grown virus showed one of the two distinct polypeptide patterns of IBV described by Nagy & Lomniczi (1979) and Collins & Alexander (1980 a, b). Strain Beaudette gave a pattern characteristic of the M type, while strain Connecticut gave a pattern characteristic of the C type. The polypeptides present in Connecticut virus were p170, gp98, pp50, gp28 and p26. Thus, the differences between the two pattern involve the mobility of both the large (gp94/gp98) and small (gp30/gp28) glycopeptides.. PMID- 6271901 TI - One functional copy of the long terminal repeat gene specifying the immediate early polypeptide IE 110 suffices for a productive infection of human foetal lung cells by herpes simplex virus. AB - The HSV-1/HSV-2 intertypic recombinant Bx1 (28-1) is heterotypic for the repeat sequences flanking the long unique region of the genome (IRL and TRL) and expresses both the immediate-early polypeptide IE 110 of HSV-1 and its functionally equivalent polypeptide IE 118 of HSV-2. The genome structures of five subclones of this recombinant and the immediate-early polypeptides they induce have been analysed. Subclone 14 lacked most of the IRL sequence, including the region from which part of the mRNA for IE 110 is transcribed, and expressed only the HSV-2 IE 118. Subclone 22 lacked almost all of TTL including the gene for IE 118 and induced only the HSV-1 IE 110. Since both subclones produced viable progeny in HFL cells it follows that expression of only one copy of the equivalent genes in TRL and IRL, here coding for the distinguishable polypeptides IE 110 or IE 118, is sufficient for successive complete cycles of virus replication. PMID- 6271902 TI - A comparison of the genomes of polioviruses by cDNA:RNA hybridization. AB - RNAs from 11 strains of the three serotypes of poliovirus were isolated from infected HeLa cells and hybridized with representative, radioactive complementary DNA (cDNA) from either poliovirus type 1 or type 2. Hybridization and thermal denaturation were analysed by hydroxyapatite chromatography, cDNA to either poliovirus type 1 or 2 is competent to detect about 70 to 80% of the sequences in RNAs from heterotypic strains and more than 02% of the RNA sequences in homotypic strains. The three polio serotype genomes appear to have diverged from each other by about 20%. Both poliovirus type 1 and type 2 representative cDNA can be used for the detection of poliovirus genomic sequences in RNA isolated from tissue. PMID- 6271903 TI - The plaque-forming factor for mink lung cells present in cytomegalovirus and herpes-zoster virus stocks identified as Mycoplasma hyorhinis. AB - Previous investigation of the ability of cytomegalovirus and varicella-zoster virus to replicate in a variety of cell lines suggested that both virus types plaqued with high efficiency in mink lung cells. However, many of the virus isolates used appeared to be contaminated with mycoplasma. We now report that the observed cytopathic effect is due to a mycoplasma which grows lytically to high titre in mink lung cells, but is difficult to cultivate in cell-free media. The mycoplasma was plaque-purified and shown to contain DNA with a buoyant density of 1.684 g/ml, with restriction endonuclease patterns identical to the porcine mycoplasma M. hyorhinis. This was confirmed by serological identification. PMID- 6271904 TI - Restriction endonuclease analysis of the DNA from varicella-zoster virus: stability of the DNA after passage in vitro. AB - DNA was extracted from nucleocapsids isolated from WI-38 cells infected with two different strains of varicella-zoster virus (VZV). The DNAs were treated with each of six restriction endonucleases (EcoRI, HindIII, Bg/I, Bg/II, Sal I and Bam HI) and small, but reproducible differences in restriction endonuclease patterns were observed. These strains were passaged in WI-38 cells and in primary guinea pig embryo (GPE) cells, followed by restriction endonuclease analysis of the DNAs. No changes were observed in the restriction prpofile of the DNA of one of the strains (VZV-KMcC) after 46 passages in WI-38 cells but small differences were observed after 72 passages. No changes were observed after 30 passages of another strain (VZV-AW) in WI-38 cells. Twenty passages of VZB-KMcC in GPE cells did result in minor alterations of its DNA. It was concluded that VZV DNA was sufficiently stable after multiple passages in WI-38 cells to make restriction endonuclease analysis a valuable epidemiological tool for strain differentiation. PMID- 6271905 TI - Biological properties and carbohydrate composition of human parainfluenza virus type 1 in two host systems. AB - Human parainfluenza virus type 1 (HA2 virus) grown either in embryonated hens' eggs or in M. rhesus monkey kidney cells showed differences in size, biological properties and carbohydrate composition. Egg-grown virus showed a larger size (233 nm versus 167 nm), a higher neuraminidase activity (specific activity and initial and maximum velocity) and a higher haemolytic activity than monkey kidney cell-grown virus. The haemagglutinin titre was identical for the HA2 strain grown in both host systems when tested with human O Rh+, guinea-pig and hen red blood cells, but reduced by more than 100-fold when tested with grivet monkey red blood cells. In addition, the carbohydrate content (mainly neutral sugars) was higher in egg-grown virus (9.2%) than in virus grown in MK cells (5.7%), and the amino to neutral sugar ratio was lower (1.2 versus 2.1). The sugars were identified as fucose, mannose, galactose, glucose, glucosamine and galactosamine. The prominent neutral monosaccharide was glucose in egg-grown virus and fucose in MK cell-grown virus. HA2 virus infection of MK cells increased fucose and glucose, and decreased mannose and galactose levels. PMID- 6271906 TI - Molecular cloning of infectious DNA from human papovavirus BK in Escherichia coli. AB - Recombinant DNA constructed from unit length BK virus DNA and from several defective viral forms was cloned in Escherichia coli HB101. The cloned unit length BK virus DNA retained its infectivity for human embryonic kidney cells, whereas the cloned defective DNA showed no infectivity. Restriction endonuclease digestion of cloned defective DNA and purified virus DNA indicated that many of the defective forms contain re-iterated sequences. PMID- 6271907 TI - Acute and recurrent herpes simplex in several strains of mice. AB - Acute and recurrent herpes simplex was studied after infection in the ear of two outbred and five inbred strains of mice. In all strains tested there was clinical evidence of infection, and a proportion of the mice became latently infected in the cervical ganglia. Six weeks after infection, when attempts were made to induce recurrent desease by stripping the ears of the mice with cellophange tape, a proportion of animals of each strain developed recurrent disease, characterized by erythema in the skin. At monthly intervals thereafter, the ears were stripped again and, on each occasion, a proportion of the animals developed recurrent disease, with the exception of Balb/c mice. The different reaction of Balb/c and other inbred strains might prove useful in studies on the mechanism of control of recurrent herpes simplex. PMID- 6271908 TI - Interference between virulent and avirulent strains of Sendai virus. AB - Interference between a virulent and an avirulent strain of Sendai virus was studied by plaque morphology and analysis of viral protein and RNA. On simultaneous infection a virulent, large plaque-forming strain (RL) was inhibited by an avirulent, small plaque-forming strain (RS). The interference was dose dependent and decreased with u.v. irradiation of RS. One infectious particle was sufficient to induce the interference. SDS-polyacrylamide gel electrophoresis showed that interference was detectable in the synthesis of viral P protein; this was abolished when RS was u.v.-irradiated. Both growth and P protein synthesis of RL was restricted by superinfection with RS when this was done within 4 h after infection of RL, but the interference decreased gradually after this period and was not detectable after 8 h. Cycloheximide prolonged the period susceptible to superinfection by RS. PMID- 6271909 TI - The regulated expression of Epstein-Barr virus. III. Proteins specified by EBV during the lytic cycle. AB - The experiments show that 30 virus-induced or virus-specified proteins were synthesized in Raji cells after superinfection with Epstein-Barr virus (EBV) derived from P3HR1 cells. Using a combination of pulse labelling, application of cycloheximide blocks at different times post-infection, treatment with amino acid analogues and inhibition of DNA synthesis it was shown that three groups of proteins appear in Raji cells after superinfection; the synthesis of the proteins in any one group appears to be coordinately regulated. Amongst the six virus induced proteins which were synthesized immediately after release from an early cycloheximide block one would expect to find those proteins essential for the transition from EBNA to EA synthesis. Using human sera with differing specificities for the various antigen groups 11 proteins were identified as being specifically precipitated by sera having high titres against the EBV-induced early antigen complex. PMID- 6271910 TI - Herpesvirus tamarinus and its relation to herpes simplex virus. AB - We have characterized Herpesvirus tamarinus (HT), a virus of New World primates related to herpes simplex virus. HT virion DNA cross-hybridized appreciably with herpes simplex virus type 1 (HSV-1) DNA. The HT-HSV-1 cross-hybridizing DNA sequences were distributed over the length of the genome; some DNA sequences, however, were apparently more homologous than others. HT virion DNA was estimated from the sum of restriction endonuclease fragments to have a mol. wt. 98 X 10(6). Only 1 molar fragments were found with some restriction endonucleases and 0.5 and 1 molar fragments were found with other restriction endonucleases but no evidence was found for 0.25 molar fragments. Independent isolates of HT from different species of South American primates gave nearly identical restriction endonuclease cleavage patterns of virion DNA. Although cross-reacting antigenic proteins of HT and HSV-1 were not detected by neutralization or immunofluorescence tests, cross reacting antigenic proteins were detected by SDS-polyacrylamide gel electrophoresis of immunoprecipitates of 35S-labelled infected cell extracts. PMID- 6271911 TI - Transfer RNAs associated with vesicular stomatitis virus. AB - The predominant RNAs in purified VSV particles are 42S and 4S in size. The 4S RNA is host transfer RNA that did not incorporate detectable radiolabel during VSV infection and was detected by in vitro labelling. Surprisingly, when BHK cells were prelabelled for 30 to 54 h before infection, the incorporation of [3H]uridine and [32P]orthophosphate into virus tRNAs remained very low and virus tRNAs were found to have a 5- to 15-fold lower specific activity than the total host tRNA, the value depending, in part, upon the period of prelabelling. Two dimensional gel electrophoresis and partial sequence analysis indicated that the virus tRNAs include most species of host tRNA and no singly predominant species was apparent. Transfer RNAs are packaged by several enveloped viruses, but we have not found 4S RNA in reovirus, which lacks an envelope. We suggest that VSV contains a membrane-associated population of tRNA which has a slower rate of turnover than the total population of cellular tRNA. PMID- 6271912 TI - An interference phenomenon associated with a measles virus SSPE isolate (Halle). AB - A measles virus (Halle SSPE isolate) induced ring plaque phenomenon (concentric rings of living and dead cells) has been shown to be associated with the temperature-dependent production of interfering particles. The interfering particles have been purified by potassium tartrate linear gradient centrifugation and have buoyant densities of 1.15 g/ml (M particle) and 1.06 g/ml (L particle) respectively. Both interfering particle population have been shown to decrease the yield of wild-type measles virus from infected cells by 50% when co-infection experiments were performed. Neither the M or L particle population interfered with the growth of VSV in the same host-cell system. PMID- 6271913 TI - Vesicular exocytosis of foot- and -mouth disease virus from mammary gland secretory epithelium of infected cows. AB - Foot-and-mouth disease virus particles were observed by electron microscopy in the cytoplasma of alveolar secretory cells of the bovine mammary gland after contact exposure of uninfected cows to pits with foot-and-mouth disease. Virus, contained in membrane-limited vesicles, was released from the basal and peranuclear portions of the cells into the intracellular and extracellular spaces by an exocytotic mechanism similar to that of the release of th milk-fat globule. Virus was released into the lumen from the apical portion of the cell both by membrane-limited vesicles and by the merocrinal exocytosis of casein-associated virus. The lytic release of virus was observed in 20% of the preparations observed. PMID- 6271914 TI - A restriction map of bovine papillomavirus type-1 DNA. PMID- 6271915 TI - Simian virus 40 causes persistent infection of muntjac cells in the absence of virus transformation. AB - SV40 infection of Muntiacus muntjak cells (ATCC, CCL: 157) resulted in abortive transformation with formation of T antigen and induction of cellular DNA replication in the absence of virus production. These cells were resistant to stable transformation by SV40 regardless of the route of infection, including microinjection of virus into cell nuclei. The present studies show that T antigen containing cells persist and that the number of T antigen-positive cells remains constant in infected cultures, which is reflected in the nearly constant fraction of T antigen-positive cells in stationary cultures and a decrease of the fraction of T antigen-positive cells in proliferating cultures. These data suggest that following cell division only one daughter cell on the average can maintain a detectable level of T antigen. The cell cycle kinetics of proliferating muntjac cultures were not altered by the abortively transforming infection, and infectious progeny was made following cell fusion with permissive CV-1 cells. It is suggested that the few copies of viral DNA that do persist are present in a "plasmid-like' state as non-integrated DNA. In this way SV40 can appear to be lost from the majority of the cells in a culture but actually be conserved in a small population of cells for a long time. In such a state a papovavirus might go undetected in vitro as well as in vitro by even the most sensitive immunological and molecular techniques. PMID- 6271916 TI - Persistent infection of Vero cells with Tacaribe virus. AB - Persistently infected cultures have been established from Vero cells surviving primary infection with Tacaribe virus (Vero-T). The growth rate and morphological characteristics of the persistently infected cells were indistinguishable from normal Vero cells. Virus release declined during the first 6 passages, a cyclical pattern was observed between passages 6 and 16, and subsequently no virus infectivity could be detected. Co-cultivation with normal RK-13 or Vero cells enhanced virus yield from virus-producing cultures of Vero-T cells (passage 15), but the addition of susceptible cells had no effect on non-producer Vero-T cultures (passage 19). Only a small proportion (less than 1%) of the persistently infected cells tested during the first 16 passages produced infectious virus. The virus released during the early stages of persistence was temperature-sensitive if grown at 40 degree C, more thermolabile at 50 degree C than parental virus, and unable to initiate a persistent infection in Vero cells. Vero-T cells consistently showed refractoriness to homotypic Tacaribe virus superinfection and a selective graded resistance to other arenavirus replication. The possible use of viral susceptibility of persistently infected cultures as marker of antigenic relationship among Tacaribe complex viruses if considered. PMID- 6271917 TI - Type-specific detection of parainfluenza viruses by enzyme-immunoassay and radioimmunoassay in nasopharyngeal specimens of patients with acute respiratory disease. AB - A four-year solid phase enzyme-immunoassay (EIA) and radioimmunoassay (RIA) techniques were applied for the type-specific detection of parainfluenza type 1, 2 and 3 virus antigens in sonicated nasopharyngeal specimens of patients with acute respiratory disease. Guinea-pig antiviral immunoglobulins as the secondary antibodies, and horseradish peroxidase-labelled swine anti-rabbit immunoglobulins (EIA), or 125I-labelled sheep anti-rabbit IgG (RIA) as the indicator antibodies. A total of 174 nasopharyngeal specimens collected by mucus extractor were tested, and the results were compared with those obtained by a routinely used immunofluorescence (IF) technique. The same number of positive specimens were achieved by the EIA and the RIA and 3/4, 4/4, and 19/20 immunofluorescence (IF) positive nasopharyngeal specimens were positive by the parainfluenza type 1, 2 and 3 immunoassays respectively. In addition, four parainfluenza type 1 and three parainfluenza type 3 virus-positive specimens were found by the immunoassays out of 146 parainfluenza IF-negative specimens. The type-specificities of the parainfluenza immunoassays were confirmed by showing that no cross-reactions occurred when purified immunizing antigens and the EIA- and RIA-positive clinical specimens were cross-tested. The results indicate that parainfluenza type specific antigens can be detected directly in nasopharyngeal specimens by the immunoassays and the preliminary findings with a small number of positive specimens suggest that these assays have a diagnostic potential which is similar or slightly better than the IF techniques. PMID- 6271918 TI - Studies on egg drop syndrome (EDS) and chick embryo lethal orphan (CELO) avian adenovirus DNAs by restriction endonucleases. AB - Restriction endonucleases EcoRI, BamHI, HindIII, BglI, BglII, HpaI and PstI recognized 48 cleavage sites in EDS adenovirus strain B8/78 DNA, whereas CELO virus DNA was cut into 61 fragments by the same enzymes. No similarity could be detected in the restriction patterns obtained from the two avian adenoviruses. The calculated mol. wt. of B8/78 DNA was 22.6 X 10(6) [about 34.2 kilobases (kb)] and that of CELO virus DNA was 28.9 X 10(6) (about 43.7 kb). The fragments generated from B8/78 DNA by EcoRI and BamHI were physically mapped. PMID- 6271919 TI - Cytomegalovirus: an ultrastructural study of the morphogenesis of nuclear inclusions in human cell culture. AB - We investigated the ultrastructural development and maturation of cytomegalovirus (CMV) nuclear inclusions (NIs) in human embryo thyroid cells at 1 to 144 h post infection. At 5 h, most cells had rounded from an initial fibroblastic appearance and contained early NIs. At 24 h, early NIs were larger and better defined. At 48 h, although early NIs were still present, most cells had larger and presumably more mature NIs. These latter NIs consisted of several subunits, each made up of a fibrillar network enclosing an electron-lucent area which contained coarse and delicate granules. Also, at 48 h, virus particles were first seen in the nucleoplasm. At 72 h, in cells with more developed NIs, virus particles were closely associated with the fibrillar network. Between 96 and 144 h, the NIs reached maximum size and were made up of numerous subunits. The results indicate that two types of NIs coexist during CMV infection. The appearance of the early the late NIs coincides with the reported peaks of CMV DNA synthesis and thus may explain the biphasic pattern of DNA synthesis in CMV infection. Morphogenetic features of the NIs conform with the hypothesis that synthesis of CMV DNA may occur in the centre in each NI subunit and that the fibrillar network represents condensing capsid proteins. PMID- 6271920 TI - Viral hepatitis: report on WHO informal consultation. PMID- 6271921 TI - Prevalence of antibodies to Hepatitis A antigen in sera from patients with haematological malignancies. AB - The prevalence of antibodies to hepatitis A (anti-HA) was investigated in patients attending a clinic for haematological malignancies. Patients were divided into three age groups; the number with antibodies in each group was recorded. Of the 130 patients studied, 44% of those aged 4-26, 70% of those aged 27-50, and 83% of those aged 51-76, were positive. This study identifies patients attending a clinic for haematological malignancies (ie, patients with immunologic deficiencies) as a group of people having a high prevalence of anti-HA. In the oldest group (51-76), urban patients had a higher prevalence than rural patients a pattern similar to that observed in elderly Red Cross blood donors in a previous study. Seroconversion was noted in six patients, three of whom seroconverted from anti-HA-negative to anti HA-positive within 12 months of the initial visit to the clinic. PMID- 6271922 TI - The persistence of papovavirus BK DNA sequences in normal human renal tissue. AB - Evidence has accumulated indicating that BK virus, following an inapparent primary infection, persists in the renal organs of normal healthy individuals and reactivates upon immunosuppression. Data to support this hypothesis are presented and suggest that BK virus DNA sequences are present at very low levels in the kidneys of more than 50% of the population and that this persistence is localized in several foci within these organs. PMID- 6271923 TI - Detection of antibody to varicella-zoster virus by competitive and IgM-antibody capture immunoassay. AB - A simple, sensitive, and specific competitive solid phase immunoassay for antibody to varicella-zoster virus (anti-VZV) is described. The assay uses reagents that can easily be prepared and is sparing of viral antigen. Using a solid phase coated with sheep IgG from a serum raised against human mu-Fc, the same reagents will accurately detect anti-VZV IgM. The competitive assay divided sera from children from adults into immune and nonimmune groups that closely correlated with a history of previous VZV illness. It was not affected by the presence or absence of antibody to other herpes viruses. The IgM antibody capture assay demonstrated the presence of anti-VZV IgM in sera from patients with both varicella and zoster and gave negative results in patients with infections unrelated to VZV and in healthy blood donors. PMID- 6271925 TI - Dual action of pentobarbitone on GABA binding: role of binding site integrity. AB - The effects of pentobarbitone on the binding of gamma-aminobutyric acid (GABA) to crude synaptosomal rat brain membranes were studied. In extensively washed P2 membranes, pentobarbitone had a biphasic action: at concentrations ranging between 12.5 and 500 microM, pentobarbitone enhanced GABA binding in a concentration-dependent manner; at concentrations greater than 500 microM, this enhancement was progressively reversed towards control levels of GABA binding. The effect of pentobarbitone seen at higher concentrations may reflect a GABA mimetic action, since similar concentrations enhanced diazepam binding to washed P2 membranes, an effect antagonized by bicuculline methochloride and picrotoxinin. When washed P2 membranes were incubated in 0.5% Triton X-100 (30 min at 37 degrees C), the enhancement of GABA binding by low concentrations of pentobarbitone was abolished, while at higher concentrations GABA binding was progressively inhibited, suggesting that the GABA-mimetic action is retained. When washed P2 membranes were subjected to high-frequency homogenization, the biphasic dose-response relationship for pentobarbitone was markedly shifted to the right. The choice of membrane preparation appears to be a critical factor in examining drug-receptor interactions in vitro, at least for those involving GABA and the barbiturates. PMID- 6271924 TI - Solubilization of collagen-tailed acetylcholinesterase from chick retina: effect of different extraction procedures. AB - We have extracted acetylcholinesterase from young chick retinas by homogenization in different solutions combining high salt concentration, ionic and nonionic detergents, and EDTA, looking for an optimum procedure for the solubilization of collagen-tailed, asymmetric structural forms of the enzyme. High salt and EDTA seem to be the only necessary requirements for the solubilization of acetylcholinesterase as the A12 form (20S), and the presence of detergent in the homogenization medium does not significantly improve the yield of tailed enzyme. Extraction in the absence of detergent has the potential advantage of a threefold enrichment of tailed enzyme, because only about one-third of the total retinal acetylcholinesterase activity is solubilized. Divalent cations, especially Ca2+, seem to be involved in the attachment of the tailed enzyme to the retinal membranes, at the tail level. High salt-EDTA-extracted 20S acetylcholinesterase (without detergent) aggregates in the presence of exogenous Ca2+ and becomes "insoluble." However, the aggregated 20S acetylcholinesterase can be completely recovered and brought back into solution by further addition of EDTA. Besides, the aggregation can be prevented by the inclusion of Triton X-100 in the homogenization buffer or by adding the detergent concurrently with Ca2+. It is postulated that the acetylcholinesterase collagenous tail is coated by acidic lipid molecules hydrophobically bound to the tail protein so that Ca2+ ionic bridges would actually link these lipid molecules (and consequently the tail) to the membrane matrix. Removal of the lipid coat (e.g., by Triton X-100) produces tailed acetylcholinesterase molecules that no longer aggregate in the presence of Ca2+ and are fully accessible to collagenase digestion. PMID- 6271926 TI - Biphasic modulation by ACTH-like peptides of protein synthesis in a cell-free system from rat brain. AB - Brain protein synthesis in a cell-free system was stimulated by 10(-8) M-ACTH1 24. This stimulatory effect was completely inhibited by aurintricarboxylic acid (ATA), an inhibitor of reinitiation of new peptide chains. The N-terminal peptide sequence 4-10 exerted a biphasic modulation of cell-free protein synthesis, i.e., a stimulation at low concentrations (10(-8) and 10(-10) M) and an inhibition at a high concentration (10(-4) M). The D-isomer, ACTH4-10-7-D-phe, also showed a biphasic modulation that, however, was in a direction opposite to that shown by ACTH4-10-7-L-phe at 10(-8) M and 10(-4) M. PMID- 6271927 TI - Nevus planus of the skin associated to jugulo-tympanicum paraganglioma. A new neurocutaneous syndrome? PMID- 6271928 TI - Myotonic muscular dystrophy. Time-dependent alterations in erythrocyte membrane fluidity. AB - The muscle cell membrane may be the site of the basic molecular defect in myotonic muscular dystrophy. Many laboratories, including our own, have suggested that this defect may also be manifested in membrane of extraneural tissue. In previous studies, we found that electron spin resonance results suggested an increased membrane fluidity in erythrocyte membranes that had aged two days in buffer, but we and others could find no such changes in fresh erythrocyte membranes. To investigate these findings further, the results of an initial study of the time course of the membrane fluidity changes in erythrocytes in myotonic muscular dystrophy are given in the present report. They suggested that increased membrane fluidity changes in erythrocytes in myotonic muscular dystrophy are given in the present report. They suggested that increased membrane fluidity in myotonic dystrophy is manifested after two days of in vivo ageing and confirm our original findings. These results are discussed in relation to possible effects of metabolic deprivation or of protein-lipid alterations in erythrocytes. PMID- 6271929 TI - Presynaptic depolarization of terminals of rubrospinal tract fibers in intermediate nucleus of cat spinal cord. PMID- 6271930 TI - Thalamic relay for group I muscle afferents of forelimb nerves in the monkey. PMID- 6271931 TI - Electrophysiological identification of mitral and tufted cells and distributions of their axons in olfactory system of the rat. PMID- 6271932 TI - Internal irradiation for cystic craniopharyngioma. AB - The authors report the results of internal irradiation with labeled chromic phosphate (32P) and gold-198 (198Au) colloid in eight cases of cystic craniopharyngiomas. They used a newly developed dosimetric formula, by which the radiation dose at the cyst wall and at any point far from the radioactive source can be calculated. Ten courses of irradiation in eight patients were carried out by injection of either 32P or 198Au colloid into the cyst through an Ommaya drainage system that had been placed at craniotomy. Follow-up studies ranging from 13 to 156 months revealed that all cysts were effectively treated, with elimination of fluid or collapse of the cyst. This was confirmed by Conray cystography and/or computerized tomography. Not only the dose delivered to the wall but also the thickness of the cyst wall and the location of the cyst are important factors in planning internal irradiation. A safe and adequate dose to the cyst wall could range between 9000 to 30,000 rads for craniopharyngioma. This treatment is suitable for large cysts that are thought to be difficult to remove radically, recurrent cysts resistant to previous treatment, or multiple cysts. Internal irradiation may also be applicable in other cystic intracranial tumors if dosimetry is calculated accurately. PMID- 6271933 TI - Reoperation for glioblastoma. AB - The results of a second operation for tumor removal in 24 adult patients with supratentorial glioblastoma multiforme or anaplastic astrocytoma were analyzed. The median survival time after reoperation was 14 weeks. Five of the 24 patients lived 6 months or longer after reoperation. Only three of these patients maintained a Karnofsky rating (KR) of at least 60 for 6 months or longer after reoperation. Preoperative neurological status (KR) is the most significant determinant of survival after reoperation (p = 0.02). When the KR is at least 60, median survival after reoperation is 22 weeks. When the KR prior to reoperation is less than 60, median survival is 9 weeks. Only one of 13 patients with a KR of less than 60 prior to reoperation survived longer than 6 months after the second operation. The interval between first and second operation is significantly related to survival (p = 0.03), but when adjustment is made for the KR the interoperative interval is no longer significantly related to survival after the second operation (p = 0.39). Age, sex, and location of tumor were not significantly related to duration of survival. This study suggests that reoperation is most likely to produce the best result when the KR is at least 60 and the interval between operations is longer than 6 months. Using these criteria, one-third of the patients could be expected to survive with a KR of at least 60 for 6 months. The study indicates that reoperation should not be carried out when the KR is less than 60. PMID- 6271934 TI - Malignant fibrous histiocytoma of the meninges. Histological, ultrastructural, and immunocytochemical studies. AB - A case of malignant fibrous histiocytoma arising from the meninges in a 12-year old boy is reported. This tumor presented as an extracerebral hemorrhagic mass in the left frontal lobe without obvious invasion of the underlying brain. Histologically, a malignant tumor with the characteristic storiform pattern of malignant fibrous histiocytoma with focal areas of hemorrhage was observed. Ultrastructurally, fibroblastic cells, undifferentiated cells, histiocytic-type cells, multinucleated tumor giant cells, and xanthomatous cells were distinctly seen. The possible cell of origin of these meningeal tumors and their prognosis are discussed. The significance of considering this entity in the differential diagnosis of those tumors of the central nervous system with a spindle-cell or xanthomatous component is discussed, and the value of ultrastructural and immunocytochemical study with glial fibrillary acidic protein in the diagnosis is stressed. PMID- 6271935 TI - Quantitation of experimental canine infarct size with multipinhole and rotating slanthole tomography. AB - Myocardial infarct size was estimated by three methods in a canine model, using Tc-99m pyrophosphate at 24 and 48 hr after coronary ligation. A gamma camera provided anterior, LAO, and lateral views, and was then fitted with multipinhole (MPH) and rotating-slanthole (RSH) collimators for tomographic studies, processed by computer to display frontal sections of the chest. Infarct weight was measured postmortem for comparison. All transmural infarcts were detected by all three imaging techniques. RSH tomography was superior to both MPH tomography and planar imaging for the detection of nontransmural infarction. Infarcts as small as 1.0 g were detected. Estimates of infarct volume measured from RSH slices showed an excellent correlation with infarct weight (r = 0.89) and were reproducible within acceptable limits. Estimates on infarct volume measured from MPH slices demonstrated a significantly poorer correlation with infarct weight (r = 0.48, p less than 0.01). Both tomographic techniques may improve infarct visualization by suppressing overlying activity and increasing contrast between infarct and background, but both produce significant blur artifacts that hamper their utilization by inexperienced observers. PMID- 6271936 TI - Marked gallium accumulation in neurogenic arthropathy. PMID- 6271937 TI - Thyroid uptake of Tc-99m compromising in vivo RBC tagging. PMID- 6271938 TI - [Location of tumor masses in epipharyngeal space and skull--application of CT scanning (author's transl)]. PMID- 6271939 TI - Phagocytic recognition of bone by macrophages. AB - There is now persuasive evidence that osteoclasts are formed by the fusion of mononuclear phagocytes. The presence of osteoclasts on bone thus infers the phagocytic recognition bone by mononuclear phagocytes. I compared the phagocytic recognition by macrophages of hydroxyapatite, and native, demineralised and deproteinated bone powder. Removal of mineral resulted in loss of phagocytic recognition by macrophages, while removal of the organic component resulted in avid binding of bone fragments to a similar degree to that observed for hydroxyapatite crystals. The organic component of bone therefore protects the hydroxyapatite crystals of the mineral component from phagocytic recognition by macrophages. A simple model for bone resorption would be that resident bone lining cells (active and resting osteoblasts) remove the organic protection under appropriate stimuli and thus allow phagocytic recognition of the underlying mineral by macrophages, which accumulate, fuse and commence digestion. PMID- 6271940 TI - Oncogenic viruses. PMID- 6271941 TI - Doppler flow and radionuclide scan studies in the evaluation and management of peripheral artery thrombosis in the neonate. PMID- 6271942 TI - Type I phosphodiesterase in the isolated, brush-border membrane of Hymenolepis diminuta. AB - The isolated, brush-border membrane of Hymenolepis diminuta contained an enzyme which hydrolyzed phosphodiester bonds. This enzyme appeared to be a Type I phosphodiesterase (E. C. 3.1.4.1) (produces nucleoside 5'-phosphates) and had no activity against synthetic, Type II phosphodiesterase substrates (mononucleotides substituted at the 3' position). The effects of various potential inhibitors of enzymatic activity, and cation requirements of this enzyme, demonstrated a distinct difference between the phosphodiesterase and alkaline phosphatase activities of the isolated, brush-border membrane. SDS-polyacrylamide gel electrophoresis of the isolated membrane preparation, followed by localization of phosphodiesterase activity in the gels, indicated the enzyme had a molecular weight of approximately 87,000. Thus, the phosphodiesterase activity represents a previously undescribed, membrane-bound enzyme of the brush-border of Hymenolepis diminuta. PMID- 6271943 TI - Evaluation of durapatite ceramic as an alloplastic implant in periodontal osseous defects. I. Initial six-month results. AB - Eight patients received a new polycrystalline ceramic form of pure hydroxylapatite (Durapatite) as a bone implant material in various types of infrabony defects following internally beveled full thickness flaps, root planing, and defect debridement. All osseous margins and defects were measured from the CEJ using a standardized periodontal probe. Similarly debrided defects that were not implanted served as controls. Defect selection as either experimental or control site was based on either split-mouth or alternating defects design. Periodontal dressing and systemic tetracycline were used for 10 days. Results for documentation and plaque control were at 10, 20, and 30 days, and 3 and 6 months. Measurements relating to defect changes were made at the 6 months surgical re-entry. For evaluation purposes original defect depths were divided into three groups. In Group I (less than 3 mm) defect fill was 1.2 mm (60%) for the implanted defects and 0.6 mm (40.5%) for the control sites (significantly different at P less than 0.05). In Group II (3--6 mm) defect fill of 1.7 mm (48.5%) for implanted sites was significantly better than the 0.1 mm (11.1%) for the control sites. In the deepest group (Group III greater than or equal to 6 mm) Durapatite placement yielded 2.6 mm (39.9%) of defect fill while debridement alone resulted in 1.3 mm (14.8%) of fill. Hard tissue changes demonstrated a substantial advantage for use of Durapatite over controls, while soft tissue changes were similar for both. The clinical impression at re-entry and the numerical data indicate that pure hydroxylapatite ceramic has a definite potential as an alloplastic implant in the treatment of periodontal osseous defects. PMID- 6271944 TI - In vitro and in vivo interactions of delta 8-tetra-hydrocannabinol and its metabolites with hepatic microsomal drug metabolizing enzyme systems of mice. AB - Interactions of delta 8-tetrahydrocannabinol (delta 8-THC) and its metabolites, 11-hydroxy-delta 8-THC, 11-oxo-delta 8-THC and delta 8-THC-11-oic acid, with hepatic microsomal drug metabolizing enzyme were studied in vitro and in vivo using mice. All the cannabinoids used were shown to produce a type I spectral change of cytochrome P-450 in hepatic microsomes. THe apparent binding affinities (Ks) for delta 8-THC, 11-hydroxy-delta 8-THC, 11-oxo-delta 8-THC and delta 8-THC 11-oic acid were 5.2, 169.6, 33.2 and 148.8 microM, respectively. delta 8-THC, 11 oxo-delta 8-THC and delta 8-THC-11-oic acid (100 microM) caused a significant stimulation of NADPH oxidation in vitro with microsomes. In addition, delta 8-THC (20, 40 and 80 microM) markedly inhibited p-nitroanisole O-demethylase and aniline hydroxylase in microsomes. 11-Hydroxy-delta 8-THC and 11-oxo-delta 8-THC also showed the inhibitory effect, but to lesser extents. Furthermore, single pretreatments with delta 8-THC and 11-oxo-delta 8-THC (30 mg/kg, i.v.) significantly decreased activities of p-nitroanisole O-demethylase and aniline hydroxylase in hepatic microsomes, accompanying a decrease of cytochrome P-450 content in case of delta 8-THC. On the other hand, all these pretreatments except for that with delta 8-THC-11-oic acid showed a stimulatory effect on p nitrophenol glucuronidation with hepatic microsomes, in contrast to an inhibitory effect in vitro. PMID- 6271945 TI - Solubility behavior of barbituric acids in aqueous solution of sodium alkyl sulfonate as a function of concentration and temperature. AB - The solubility of 13 barbituric acids was determined in aqueous solutions of sodium alkyl sulfonate. The effects of concentration and temperature were investigated, and the thermodynamic functions of the solubilization process were calculated. An analysis of the location of a solubilized species within a micelle is suggested in terms of the sign and amplitude of the standard entropy of solubilization, which is strongly positive for micelle penetration and negative for adsorption. A solubilization mechanism through adsorption onto the micellar surface is suggested for most of the barbituric acids studied. The enthalpy/entropy compensation phenomenon was identical for barbituric acids in ionic and nonionic (polyoxyethylene lauryl ether )surfactant solutions with a compensation temperature of 270 degrees K, indicating common behavior of these compounds with respect to micellar solubilization. The concept of molecular surface area was used to correlate the free energy of solubilization of the solutes to their size and structure. A linear relationship was found with an excellent correlation factor for the alkane derivatives of the 5-ethyl-barbituric acids. The specific behavior of some of the barbituric acids investigated is discussed. PMID- 6271946 TI - Polymorphism of spray-dried microencapsulated sulfamethoxazole with cellulose acetate phthalate and colloidal silica, montmorillonite, or talc. AB - Sulfamethoxazole was microencapsulated with cellulose acetate phthalate and talc, colloidal silica, or montmorillonite clay by a spray-drying technique. The surface topography of the products varied with the type of excipient used and the pH of the suspending medium. The products without the excipient were coated with flake-like crusts, while the products containing the excipient tended to become well-rounded spheres. In addition, the crystalline form of sulfamethoxazole converted from Form I to an amorphism and Form II during the spray-drying process. This polymorphic transformation was attributed to the interaction of cellulose acetate phthalate with sulfamethoxazole. Increasing the concentration of cellulose acetate phthalate in the formulation increased the attainment of amorphism. Form II was also obtained by freeze and vacuum drying. Talc was the only excipient that contributed to polymorphism, which occurred in the alkaline suspension medium. Montmorillonite products prepared from the acidic medium exhibited an exothermic differential scanning calorimetry thermogram, which might be interpreted in terms of adsorption of the fused sulfamethoxazole with the internal surface of montmorillonite, PMID- 6271947 TI - Modulation of sodium channels of squid nerve membranes by grayanotoxin I. PMID- 6271948 TI - The effect of dietary lipid on The binding of [3H] dihydroalprenolol and adenylate cyclase activity in rat atria. PMID- 6271949 TI - [Roentgenologic aspects of malignant fibrous histiocytomas of soft tissues and of bone (author's transl)]. AB - The histiocyte, macrophage or tissue monocyte is a cell of the mononuclear system. The histiocytes have the abilities to phagocytose by utilizing hydrolytic enzymes they have synthesized, and which can synthesize specific proteins to enhance immune responses. The malignant fibrous histiocytoma is a relatively common neoplasm of the mesenchymal tissues derived from histiocytes capables of wide latitude in the expression of differentiation and pleomorphic appearance. This tumor arises as a primary tumor of soft tissue as well as of bone. Four of ten cases of malignant fibrous histiocytomas are documented. This report adds to the understanding of the entity of the tumor in that the diagnosis of malignance was made using roentgenologic criteria and confirmed by biopsy and by autopsy. The most commun locations were of the extremities and extensive local recurrence and metastases are the characteristics. The purpose of this study is to analyse the usefulness the roentgenologic methods in the diagnosis of the malignant histiocytoma: conventional radiography, radionuclide scanning, computed tomography, sonography, xeroradiography, angiography. The differential diagnosis is reported. An algorithmic roentgenologic approach for the evaluation of the malignant fibrous histiocytoma is proposed. PMID- 6271950 TI - Fluorescence visualization of receptor-bound low density lipoprotein in human fibroblasts. PMID- 6271951 TI - Kidney membrane binding of native parathyroid hormone compared to binding of its synthetic 1 - 34 fragment. AB - Kidney membrane binding of tritiated native parathyroid hormone (PTH) was compared to that of its active fragment 1 - 34 PTH. Native hormone specific binding is transient and disappears rapidly (integral of 30 minutes) at 37 degrees C but is stable up to 2 hours at 0 degrees C. The rate of binding loss appears relatively independent of the amount of hormone-receptor complex or of the amount of cold PTH in the medium. Loss of specific binding is also seen with iodinated PTH. Loss of specific binding of native PTH does not appear to be the result of enzymatic degradation of the hormone since significant amounts of intact hormone are present, both bound to membranes and in the medium after incubation. Biologically active tritiated 1 - 34 PTH binds specifically to isolated membrane, and both total and specific binding is stable for at least one hour at room temperature. The pH dependence for binding of 1 - 84 PTH and its activation of adenylyl cyclase are very similar, but differ from that reported for specific binding of 1 - 34 PTH. these results suggest that the interaction of native PTH with kidney cells may be more complex than that of its 1 - 34 fragment. PMID- 6271952 TI - (3H)-vasopressin binding to isolated rat hepatocytes and liver membranes: regulation by GTP and relation to glycogen phosphorylase activation. AB - Specific vasopressin binding to rat hepatocytes and rat liver membranes was measured using biologically active (3H)-Tyr2-Lys8-vasopressin (8.5 Ci/mM). In both systems, vasopressin binding was found to be time-dependent, reversible, and saturable. The kinetic parameters for vasopressin binding were: apparent dissociation constants (Kd): 4.9 nM and 15 nM; maximal binding capacities: 0.83 pmoles/mg protein and 2.10(5) sites/Cell for purified membranes and intact cells respectively. The relative affinities of 19 vasopressin structural analogues were deduced from competition experiments and compared to the previously determined glycogenolytic (or antiglycogenolytic) potencies of these analogues. For both agonists and antagonists, a highly significant correlation was demonstrated between pKd and pKa (or pKi) values, suggesting that the detected binding sites are the physiological receptors involved in the glycogenolytic action of vasopressin on the rat liver. The affinity of antagonists for binding to these receptors is the same for both membranes and cells. In contrast, agonists which bind to vasopressin receptor sites have a higher affinity for purified membranes than for intact cells (Kd membranes/Kd cells = 8 +/- 1). GTP (0.1mM) reduced the affinity of agonists but not of antagonists for binding to membranes and abolished the differences between Kd values for binding to hepatocytes and membranes. PMID- 6271953 TI - Calcium binding to plasma membranes from normal and SV40 transformed hamster lymphocytes. AB - The calcium binding properties of isolated plasma membranes from normal and SV40 transformed hamster lymphocytes were compared over the Ca2+ concentration range of 10(-5)M to 5 x 10(-3)M and at physiological ionic strength. At all Ca2+ concentrations, normal membranes bound more Ca2+ than tumor membranes; at blood Ca2+ levels (1-2 mM) plasma membranes of normal cells bind twice as much as membranes from tumor cells. Normal plasma membranes demonstrated positive cooperative Ca2+ binding whereas tumor membranes displayed non-interacting Ca2+ binding sites. Ca2+ binding to both membranes was insensitive to Mg2+ (0.1 to 2.5 mM). A pH shift from 7 to 6 resulted in a 70% decrease of normal membrane-bound Ca2+ compared to a 40% decrease observed with tumor membranes. Extracellular surface Ca2+ binding to intact cells was also studied after a 72-hour equilibration of cells with 45 Ca2+ and with ethyleneglycol-bis-(beta-amino-ethyl ether) N,N'-tetraacetate chelation as marker for surface Ca2+. Tumor cell surface Ca2+ binding was only 10% of that observed with quiescent lymphocytes. Normal lymphocytes stimulated to divide with phytohemagglutinin also showed a decreased level of surface Ca2+ (50%). However, plasma membranes isolated from non-dividing and phytohemagglutinin-stimulated lymphocytes exhibited equivalent Ca2+ binding. PMID- 6271955 TI - Cytosol preparations are inadequate for quantitating unoccupied receptors for 1,25-dihydroxyvitamin D3. PMID- 6271954 TI - Uncoupling of the glucagon receptor-adenylate cyclase system by glucagon in cloned differentiated rat hepatocytes. AB - The ability of glucagon to induce a state of desensitization to glucagon responsiveness has been examined in a cloned line of normal, differentiated, diploid rat hepatocytes (RL-PR-C). These cells, which respond to glucagon with increased production of cyclic AMP, become refractory to further stimulation of cyclic AMP synthesis following a 4 hour exposure period of the cells to the hormone. Refractoriness to glucagon was demonstrated over a wide range of hormone concentrations (10(-12) to 10(-6) M). In desensitized cells that were subsequently washed free of the hormone, recovery from refractoriness was complete by 20 hours. The mechanism underlying this desensitization does not appear to involve decreased receptor numbers, increased efflux of cyclic AMP from the cells, increased degradation of cyclic AMP by phosphodiesterase, or an alteration in the catalytic unit of the adenylate cellular cyclic AMP responsiveness to glucagon in normal RL-PR-C hepatocytes may involve glucagon a reversible uncoupling of glucagon receptors from adenylate cyclase. In addition, late passage, spontaneously transformed RL-PR-C hepatocytes were found to exist in a state in which glucagon receptors are permanently uncoupled from adenylate cyclase. PMID- 6271956 TI - The use of ascorbate as a probe of opioid receptor structure: evidence for two independent mechanisms of receptor destruction by ascorbate. AB - Ascorbate (20mM) pretreatment of brain membrane suspensions at 37 degrees produced a rapid irreversible loss of specific opioid binding. There was no reduction in specific 3H-halo-peridol binding. Ascorbate induced loss of opioid binding under these experimental conditions was not blocked by low concentrations of EDTA or MN++. In contrast, the slowly developing loss of opioid binding during exposure to 1 mM ascorbate at 23 degrees was completely inhibited by 10(-5)M EDTA or Mn++. At 37 degrees, D-isoascorbate, and several other reducing agents (glutathione, dithiothreitol, cysteine) produced a loss of opioid binding similar to that seen with ascorbate. It is concluded that 1 mM ascorbate at 23 degrees, and 20 mM ascorbate at 37 degrees, destroy opioid binding sites by two independent mechanisms. Lipid peroxidation is implicated at low ascorbate concentrations; a reductive process appears to be responsible for the ascorbate induced loss of binding at higher concentrations. PMID- 6271957 TI - Prolactin binding in rat Langerhans islets. AB - Membrane preparations of collagenase-dispersed Langerhans islets of female Wistar rats exhibit specific binding sites for 125I-labelled ovine prolactin (125I oPrl). Almost negligible binding was detected in islets of male animals. The binding is a saturable and time-temperature dependent process, equilibrium being reached after 16 h incubation at 0 degrees C. The bound oPrl is not displaceable by hFSH, hLH, bGH or hGH. In contrast with other cell fractions, the 12,000 g pellet accounts for more than 80% of the specific binding of 125I-oPrl. Scatchard plots of data obtained in saturation studies indicate a single class of binding sites with Ka = 0.21 x 10(10)M-1. Protein and phospholipid moieties are essential for the receptor activity, since after trypsin or phospholipase C digestions marked loss of binding was verified. In islets of streptozotocin diabetic rats a marked reduction in the number of binding sites was observed. These findings may suggest that some of the actions of prolactin on endocrine pancreas could be explained by its specific interaction with islet cell membranes. PMID- 6271958 TI - Cell density modulates receptor-mediated internalization of nerve growth factor in pheochromocytoma cells. AB - Binding and fate of the nerve growth factor (NGF) in pheochromocytoma cells (clone PC12) have been measured with the use of iodine-labeled ligand and with 125I-NGF antibodies. With such double approach it is possible to distinguish between surface bound and total NGF bound to PC12 cells. It is found that NGF receptor complexes undergo down-regulation. This process is noticeable at low but not at high cell densities, and only in PC12 cells never exposed to NGF. Previous incubation with growth factor leads to the disappearance of down-regulation of NGF-receptor complexes. Assuming that this process is an indirect measure of NGF receptor internalization, it is concluded that it is modulated by the cell density or by previous exposure to the factor. These findings are postulated to be relevant to the mechanism of action of NGF and to its multiple effects on target cells. PMID- 6271959 TI - Opiate receptor sites in the rabbit cerebellum: autoradiographic distribution. AB - The distribution of the [3H] diprenorphine binding sites in the rabbit cerebellum has been analyzed after specific labelling of tissue slices in vitro followed by autoradiographic processing. High, moderate and very low silver grain densities were observed over the molecular, granular and white layers, respectively. Within each layer, the distribution of the autoradiographic grain exhibited no evident patterning. Therefore, further work using complementary techniques is needed to determine the precise location of the opiate receptor sites in the rabbit cerebellum. PMID- 6271960 TI - Benzodiazepine receptors in rat brain: action of Triton X-100 and localization in relation to the synaptic region. AB - Membranes were isolated from the cerebral cortices of rat brain and submitted to binding with 3H-flunitrazepam (3H-FNZP). Specific binding gave a hyperbolic saturation curve reaching a maximum between 10 - 12 nM suggesting a single population of sites. By Scatchard analysis a KD = 4 nM and a Bmax = 407 fmol/mg protein was obtained; the Hill number was 0.97. The effect of Triton X-100 was analyzed between concentrations of 6.4 x 10(-8) and 5 x 10(-1)%. With 6.4 x 10( 6) and 6.4 x 10(-3)% there is an activation of the binding without loss of protein. This activation is not reversed by washing. At 6.4 x 10(-2)% or higher concentrations there is an inhibition that is partially reversed and also solubilization of some receptors. The activation with low detergent is due to an increase in Bmax without change in KD. With 2 x 10(-1)% and 5 x 10(-1)% Triton X 100 there is considerable decrease in Bmax and in th latter case, an increase in affinity, as well. The results obtained through the action of Triton X-100 suggest that a proportion of benzodiazepine receptors are localized presynaptically. There findings are discussed in relation to previous studies from this laboratory on the localization of other central receptors with reference to the synaptic region. PMID- 6271961 TI - Effect of an LHRH analog on growth and hormone receptor levels in DMBA-induced mammary tumors in the rat. AB - Dimethylbenz(a)anthracene (DMBA)-induced mammary tumors in the rat are well known to be hormono-dependent. Daily injections of an LHRH agonist, [D-Ala6, des-Gly NH2(10)]LHRH ethylamide (LHRH-A), 1 microgram daily, for 38 days results in a 35% decrease in the number of tumors present at the beginning of the experiment compared to a decline of 45% after ovariectomy and of 8% in the control group. This is accompanied by a marked reduction in ovarian LH and FSH receptors. LHRH-A treatment also resulted in reduction in the number of progesterone and prolactin receptors in the tumors. In addition, an increase in plasma LH and FSH and a decline in plasma prolactin (PRL) concentrations are observed. The mechanisms by which the LHRH agonist induces its antitumoral effect probably relate to an ovarian desensitization to LH and FSH with a concomitant decrease in circulating levels of estrogen and prolactin, two well known stimuli for the growth of DMBA induced mammary tumors. PMID- 6271962 TI - On the dynamics of abrin binding to receptor sites in normal and Epstein Barr Virus transformed lymphocyte cell cultures. AB - The effects of treatment with varying doses of abrin, a D-galactose binding lectin, on DNA an protein synthesis of normal and Epstein Barr Virus transformed lymphocytes has been investigated. Activation, stimulation, and relative toxicity factor indices are studied, as well as possible relationships between DNA and protein synthesis rates, as measured by simultaneous tritiated thymidine (3H-TdR) and 14C-leucine uptake. Studies of the two new indices, the metabolic self and cross coupling indices lead to the prediction that there are three morphologically distinct subpopulations of EBV-transformed lymphocytes with different abrin receptor site concentrations. This prediction is supported by SEM morphological differences. Using data on EBV-transformed lymphocyte cell density as a function of time and dose of abrin, one can demonstrate that the mean number of receptors bound-EBV-lymphocyte needed to exert a biological influence lies in the interval 59,264 receptors/cell to 370.040 receptors/cell. Using a simple packing model, one can demonstrate that a theoretical estimate places the number of binding sites between 57,600 receptors/cell and 360,000 receptors/cell. PMID- 6271963 TI - Water soluble receptors for human growth hormone from rabbit liver. AB - Soluble receptors that bind human growth hormone have been prepared by incubation of liver membranes form pregnant female rabbits in 1 mM Tris buffer (ph 7.5 or 9.0) at 4 degrees C. Up to 29% of the growth hormone binding sites could be solubilized within 48 hours. The kinetics of binding of human growth hormone to the soluble receptor, the hormonal specificity and the binding parameters calculated by Scatchard analysis (Ka 2.2 x 10(9) M-1, capacity 409 fmole/mg) were essentially unchanged compared with those for the parent membrane-associated (particulate) receptor. Gel filtration on Ultrogel AcA22 indicated that the major binding peak eluted at a molecular weight of 300,000 daltons. Specificity studies showed that the soluble binding sites had a moderately high affinity for ovine prolactin (Ka integral of 1 x 10(8) M-1), but negligible affinity for insulin. Although aqueous extraction gives a lower yield of binding sites for human growth hormone than detergent extraction, it nevertheless avoids some of the problems associated with use of detergents and should facilitate the subsequent purification of the receptor in a relatively unaltered state. It may also have applicability for solubilization of other hormone receptor systems. PMID- 6271964 TI - Effect of hCG, cAMP and FSH on steroidogenesis by human corpora lutea in vitro. AB - Human corpora lutea of various ages were minced and incubated in the presence of hCG (10 i.u./ml), cAMP (10 mM) or FSH (20 mu/ml) and production of progesterone and oestradiol was measured. Cyclic AMP and hCG stimulated progesterone and oestradiol production during at least the mid- and late luteal phases, but FSH stimulated only oestradiol production during the early and mid-luteal phases and had no effect on progesterone production. This demonstrates that progesterone and oestradiol synthesis by the human corpus luteum can be independently controlled. PMID- 6271965 TI - Changes in DNA, cyclic nucleotides and steroids during induced follicular atresia in the hamster. AB - Atresia was induced in antral follicles of hypophysectomized PMSG (30 i.u.) treated hamsters by an i.p. injection of PMSG antiserum. Mature antral follicles were isolated and incubated for 1 h. When expressed per follicle, cAMP increased 108% above control levels at 2 h, while cGMP rose 117% at 4 h; both nucleotides then declined (12-72 h). DNA per follicle fell below control values by 24 h and reached its lowest point at 48 h. Serum concentrations of oestradiol-17 beta and testosterone were 50% lower by 1 h after injections of PMSG antiserum. Oestradiol and testosterone in serum, incubation medium and tissue dropped rapidly to reach basal values by 12 h. Serum progesterone showed a sharp increase at 4 h then dropped below control levels by 12 h and remained constant thereafter. In tissue and medium, progesterone rose to peak values at 8 h, then dropped to control levels with no significant changes for the remaining 64 h. The precipitous decreases in both oestradiol and testosterone in the serum, medium and tissue indicate inhibition of steroid synthesis at some point after progesterone formation. These results demonstrate the usefulness of this model for following atretic changes in a synchronized population of Graafian follicles. PMID- 6271966 TI - A gouty family with increased phosphoribosylpyrophosphate synthetase activity: case reports, familial studies, and kinetic studies of the abnormal enzyme. AB - Two male patients with urate overexcretion and clinical gout in a family showed activity of phosphoribosylpyrophosphate (PRPP) synthetase in erythrocyte lysates (3.1-fold) greater than that found in normal subjects. Hemolysates from 5 female persons in this family contained (2.7-fold) increased enzyme activity suggesting X-linked dominant transmission of the abnormality. Increased maximal velocity of the enzyme, aberrant protein pattern in polyacrylamide electrophoresis, and increased thermolability in purified enzyme suggested that this enzyme is a mutant one. From these findings, it was assumed that the characteristics of this enzyme were different from 4 previously reported enzymes. PMID- 6271967 TI - Leukotriene B4 and neutrophil function: a review. PMID- 6271968 TI - Clustering of hepatitis B virus infection and hepatocellular carcinoma in a family. PMID- 6271969 TI - Construction and properties of a recombinant plasmid containing gene 32 of bacteriophage T4D. PMID- 6271970 TI - Structure of polyoma virus late nuclear RNA. PMID- 6271971 TI - A new dominant hybrid selective marker for higher eukaryotic cells. PMID- 6271972 TI - Detection of methylated sequences in eukaryotic DNA with the restriction endonucleases Smai and Xmai. PMID- 6271973 TI - Replicon fusions promoted by the inverted repeats of Tn5. The right repeat is an insertion sequence. PMID- 6271974 TI - Synthesis, cleavage and sequence analysis of DNA complementary to the 26 S messenger RNA of Sindbis virus. PMID- 6271975 TI - Predominant end-products of prophage Mu DNA transposition during the lytic cycle are replicon fusions. PMID- 6271976 TI - Calcium-binding rate and capacity of cardiac sarcoplasmic reticulum. PMID- 6271977 TI - Cellular manganese uptake by the isolated perfused rat heart: a probe for the sarcolemma calcium channel. PMID- 6271978 TI - Calcium and calcium slow channel antagonists on cyclic nucleotide levels in the isolated rat heart. PMID- 6271979 TI - Phosphoproteins in Dictyostelium discoideum. AB - The phosphoproteins of Dictyostelium discoideum were compared at different stages of development of polyacrylamide gel electrophoresis. Certain phosphoproteins of vegetative amoebae were conserved while others appeared and disappeared during development. Four major phosphoproteins with apparent subunit molecular weights of 50,000, 47,000, 38,000, and 34,000 disappeared precociously in response to exogenous of cAMP. Two membranal phosphoproteins, with apparent subunit molecular weights of 80,000 and 81,000, appeared precociously in response to added cAMP. One of these phosphoproteins, molecular weight 80,000, has been identified tentatively as the "contact site A"glycoprotein. Another membranal protein, with apparent subunit molecular weight of 42,000, unaffected in its appearance by cAMP, has been identified tentatively as phosphoactin. PMID- 6271980 TI - Stimulation of DNA polymerase alpha by a nuclear DNA/protein complex. AB - A nuclear DNA complex containing DNA polymerase and SV40 T-antigen was isolated from nuclei of SV40-transformed mouse fibroblasts. DNA polymerase could be separated from the complex. The remaining DNA/T-antigen-containing complex stimulated DNA polymerase alpha activity about 10-fold. The complex contained 4 major proteins with molecular weights of 46, 54, 76, and 94 kilo-dalton (KD). The stimulation activity was retained by protein A-Sepharose loaded with specific IgG from SV40-tumor bearer serum, or from antisera against the 94 KD and 76 KD components and was partially inhibited in the presence of these antisera. The stimulation activity was completely abolished by treatment of the complex with trypsin or DNase I. PMID- 6271981 TI - The repair of DNA damage: recent developments and new insights. AB - This brief review presents the salient features of new developments in the enzymatic repair of base damage to DNA. DNA glycosylases and apurinic/apyrimidinic (AP) endonucleases are reviewed and evidence is presented that in at least two prokaryote systems incision of UV-irradiated DNA occurs by the sequential action of these two classes of enzymes. In contradistinction, the uvrA, uvrB, and uvrC gene products of E coli appear to function as a multi protein complex that catalyzes hydrolysis of phosphodiester bonds in damaged DNA directly. The inducible rapid repair of O6-methylguanine in E coli is also reviewed. PMID- 6271982 TI - The transforming sequences of avian myelocytomatosis virus (MC29). AB - Avian myelocytomatosis virus (MC29), a defective acute leukemia virus, has a broad oncogenic spectrum in vivo, and transforms fibroblasts and hematopoietic target cells in vitro, We have used recombinant DNA technology to isolate and characterize the sequences that are essential in the transformation process. Integrated MC29 proviral DNA was isolated from a library of recombinant phage containing DNA from the MC29-transformed nonproducer quail cell line Q5. The cloned DNA was analyzed by Southern blotting of restriction endonuclease digests and by electron microscopic visualization of R-loops formed between the cloned DNA and MC29 or helper virus RNA. It was found that the 9.2 kb cloned DNA insert contains approximately 4 kb of viral sequences and 5.2 kb of quail cellular sequences. The viral sequences contain all of the MC29 specific sequences and 5' helper related sequences as well as part of the envelope region. The size of the cloned EcoRI fragment is the same as that of the major band in EcoRI-cleaved Q5 DNA that hybridizes to viral sequences. Transfection of the cloned DNA into NIH 3T3 cells revealed that the MC29-specific sequences are functional in that they induce foci of transformed cells with high efficiency. PMID- 6271983 TI - Synthesis of lipids in mouse brain cell cultures during development. AB - Several metabolic activities in dissociated cultures of newborn mouse brain were compared to the situation in vivo. The developmental activity pattern of cerebroside-sulfotransferase, cyclic nucleotide phosphohydrolase, and beta hydroxy-beta-methyl glutaryl-coenzyme A-reductase and the synthesis and deposition of sulfatide and cholesterol in culture were estimated. The enzyme activity patterns in vivo and in culture are the same. Since the cultures show very little myelin formation, the parallel increase of enzyme activities necessary for myelination in vivo and in culture suggest the existence of intrinsic factors regulating the biochemical differentiation. In addition, the formation of the products, determined in culture, follows the patterns of the enzyme activities. Dissociated brain cell cultures are therefore a valid model for the study of biochemical parameters related to the synthesis of brain lipids during development. PMID- 6271985 TI - Effect of denervation on cyclic nucleotide metabolism in different types of skeletal muscle of the rat. AB - Slow-twitch soleus and fast-twitch extensor digitorum longus muscles of the rat were denervated unilaterally by sciatic nerve section at mid-thigh level. Activities of adenylate cyclase, guanylate cyclase, low Km and high Km cyclic AMP phosphodiesterase, and cyclic GMP phosphodiesterase were compared on the same, freshly prepared homogenates of denervated and shamoperated contralateral muscles one, two, three, or five days after surgery. As an early consequence of denervation, cyclic AMP metabolism was differentially affected in these different types of skeletal muscle. The adenylate cyclase activity of soleus muscle increased significantly by the second day following denervation and continued to rise through the fifth day, while this enzyme did not increase in denervated extensor digitorum longus even by the fifth day. The high Km cyclic AMP phosphodiesterase was already increased by day one in the denervated soleus, but not until the fifth day in the denervated extensor digitorum longus. Parallel increases beginning the first day were observed for the low Km cyclic AMP phosphodiesterase in both muscles. Since the activity of cytosolic cyclic AMP dependent protein kinase of soleus muscle was also increased two days following denervation, the changes in cyclic AMP synthetic and degradative enzymes apparently result in a rise in intracellular cyclic AMP concentration. Alterations of the cyclic GMP enzymes following denervation were similar in the soleus and extensor digitorum longus, but were delayed relative to the increases in activity in the cyclic AMP enzymes. PMID- 6271984 TI - Induction of neurite outgrowth in the IMR-32 human neuroblastoma cell line by nerve growth factor. AB - The nerve growth factor protein (NGF) stimulates neurite outgrowth form embryonic sensory ganglia and sympathetic ganglia at all stages of development. In addition, NGF is required for the maintenance of the differentiated state in adult sympathetic ganglia. A clonal cell line, IMR-32, derived from a human neuroblastoma was found to contain a population of cells that respond to NGF by exhibiting morphological differentiation. The effect of NGF on these cells is compared with that of other agents known to induce differentiation of IMR-32, including glioma-conditioned media. PMID- 6271986 TI - Protective effect of taurine on the light-induced disruption of isolated frog rod outer segments. AB - Isolated frog rod outer segments (ROS) incubated in a Krebs-bicarbonate medium, and illuminated for 2 h, show a profound alteration in their structure. This is characterized by distention of discs, vesiculation, and a marked swelling. The light-induced ROS disruption requires the presence of bicarbonate and sodium chloride. Replacement of bicarbonate by TRIS or HEPES protects ROS structure. Also, substitution of sodium chloride by sucrose or choline chloride maintains unaltered the ROS structure. Deletion of calcium, magnesium, or phosphate does not modify the effect produced by illumination. An increased accumulation of labeled bicarbonate and tritiated water is observed in illuminated ROS, as compared with controls in the dark. The presence of taurine, GABA, or glycine, at concentrations of 5-25 mM, effectively counteracts the light-induced ROS disruption. Taurine (25 mM) reduces labeled bicarbonate and tritiated water levels to those observed in the dark incubated ROS. PMID- 6271987 TI - Membrane properties of a human neuroblastoma II: Effects of differentiation. AB - The SK-N-SH cell line is a human neuroblastoma which when grown under standard culture conditions remains relatively undifferentiated. The undifferentiated SK-N SH cells are relatively inexcitable: they show only partial active responses to injections of current pulses and lack the depolarizing component of the action potential generating mechanism [Kuramoto et al, 1977]. In this study we report on the membrane properties of two subclones of the SK-N-SH, the SK-N-SH-IN (referred to as IN) and the SK-N-SH-Sy5Y (referred to as 5Y) which exhibit extensive morphological differentiation when grown with 1mM dibutyryl cAMP. Fully differentiated IN and 5Y cells have higher resting membrane potentials, in the range of -50 to -80 mV, and higher input resistance and time constants than do the undifferentiated SK-N-SH parental cell line. After 1 week in culture the differentiated IN and 5Y cells exhibit spike potentials in response to injection of current to the cell body. The presence of the Na2+-dependent depolarization was verified directly by the use of tetrodotoxin (TTX) and indirectly in experiments where veratridine (0.1 mM) markedly enhanced the influx of 22Na+. Taken together, the data indicate that generation of action potentials in these human neuroblastoma cells is to a large extent a function of morphological differentiation. PMID- 6271988 TI - Taurine binding to membranes from rat brain regions. AB - [3H]-taurine binding to membranes from different regions from rat brain was studied. Binding to membranes from cerebral cortex and its subcellular fractions, hypothalamus, olfactory bulb and cerebellum was measured. Binding to membranes from dorsal root ganglion was also determined. Na+-dependent taurine binding was consistently observed in all the membranes except those from dorsal root ganglion. A KD = 4.06 muM was obtained for binding to membranes from cerebral cortex. Na+-dependent taurine binding was displaced by 20 muM strychnine or bicuculline. Na+-dependent taurine binding with properties corresponding to a postsynaptic interaction could not be detected in any of the regions studied. The possibility of Na+-dependent taurine binding, representing binding to uptake sites or to postsynaptic receptors for GABA and glycine, is discussed. PMID- 6271989 TI - 3H-muscimol binding in synaptosomal fractions from bovine and developing rabbit retinas. AB - Bovine and rabbit retina synaptosomal fractions bind the potent and specific GABA agonist, [3H]-muscimol with high affinity and limited capacity. The high degree of pharmacological specificity and the subcellular distribution of these binding sites are similar to those reported for [3H]-GABA binding sites. These observations suggest that these sites represent the recognition sites for GABA receptors. The specific binding of [3H]-muscimol in retinal homogenates from different aged rabbits reveal a distinct developmental profile with a fivefold to six fold increase in binding between days 5 and 13. Thus, it appears that GABA receptor development continues after eye opening in rabbits (day 9-10) and that receptor maturation is delayed by at least two to three days with respect to published of GABA uptake and evoked release. PMID- 6271990 TI - Ethanol and cycloheximide alter protein and RNA synthesis of Cox astrocytoma cells in culture. PMID- 6271991 TI - Hepatic recognition and catabolism of serum glycoproteins. PMID- 6271992 TI - Perirectal infections in patients with small cell lung cancer. AB - Fifteen anorectal infections occurred in 6.4% of 188 intensively treated patients with small cell bronchogenic carcinoma. Granulocytopenia was present at onset in 13 episodes. In eight episodes, perirectal pain preceded any objective findings by two to 11 days. Ten infections resolved with recovery from granulocytopenia, but the five abscesses required incision. Septicemia accompanied four episodes; one patient died. Early recognition and therapy of this potentially fatal infection will become increasingly important as more patients with solid tumors receive intensive therapy. PMID- 6271993 TI - Pions, ions, neutrons, protons "expanding scope of radiotherapy'. PMID- 6271994 TI - Studies on the Na+-K+-ATPase in myocardial infarction. AB - Changes in the cardiac sarcolemma in myocardial infarction were studied by both determination of Na+-K+-ATPase activity and SDS gel electrophoretic analysis of sarcolemmal proteins in the canine heart. Ninety minutes after coronary ligation, Na+-K+-ATPase activity in ischemic myocardium was decreased significantly to approximately 36% of that of non-ischemic myocardium, and it remained at the lower level for 28 days. By SDS gel electrophoresis, reduction of the protein band with molecular weight of 111,000, which is suggestive of the main component of ATPase, was observed simultaneously with the reduction of Na+-K+-ATPase activity. These results indicate that ischemia for 90 minutes produces substructural changes in the sarcolemma indicating irreversible myocardial changes. PMID- 6271995 TI - Histochemical study on 5'-nucleotidase in the cerebrovascular system in stroke prone spontaneously hypertensive rats. AB - 5'-Nucleotidase activity in the cerebrovascular system in stroke-prone spontaneously hypertensive rats was histochemically investigated. In the hypertensive rats, an enhancement of 5'-nucleotidase activity was observed already in the early stage of hypertension and the enzyme activity increased with advancing age. The enzyme activity appeared earlier and stronger in the larger arteries in the basal portion than the pial arteries in the convexity of the brain and arterioles in the brain parenchyma. The activity in the arterial walls proved to be particularly strong in the thickened parts showing cellular hyperplasia, mainly at the branching portions. The reaction products localized along the plasma membrane, and also in the cytoplasma in some parts where the activity was strong. Relationship between the increased 5'-nucleotidase activity and aging of the cells composing the cerebrovasculature resulted from an accelerated or repeated cell proliferation was discussed. PMID- 6271996 TI - Purification of tonin by chromatography using soybean trypsin inhibitor coupled CH-sepharose 4B. AB - Tonin was purified from rat submaxillary gland homogenates by affinity chromatography on soybean trypsin inhibitor coupled CH-Sepharose 4B and two additional steps of conventional chromatography. The use of affinity chromatography by soybean trypsin inhibitor coupled CH-Sepharose 4B permits a new approach in the purification of tonin, since it can completely separate in one step troublesome contamination of the enzymes which showed tosyl-L-arginine methyl ester hydrochloride esterase activity. Tonin is purified 11-fold to a homogeneous state on polyacrylamide gel electrophoresis at a yield of 21%. PMID- 6271997 TI - Changes in cardiac beta-adrenoceptor concentrations in spontaneously hypertensive and experimental renal hypertensive rats. AB - Cardiac beta-adrenoceptors were studied in membrane fractions from spontaneously hypertensive rats (SHR) and rats with two-kidney, one clip hypertension (2K, 1C HT), using radioligand binding method. beta-Adrenoceptor concentration measured by [3H]-dihydroalprenolol (DHA) binding was significantly lower in cardiac membranes from two months old SHR than those from Wistar-Kyoto rats (WKY) (38.2 +/- 2.6 vs 45.1 +/- 1.8 fmol/mg protein, means +/- SEM, p less than 0.05). Cardiac membranes from 2K, 1C HT rats had also a lower concentration of beta adrenoceptors than those from the sham-operated control rats at a week after operation (30.9 +/- 2.2 vs 47.8 +/- 1.6 fmol/mg protein, p less than 0.01). But receptor affinity remained unchanged. These reduced concentrations of beta adrenoceptors were restored to control levels at 12 months old in SHR and at 6 weeks after operation in 2K, 1C HT rats, although age-dependent decrease in beta adrenoceptor was observed. The decrease in beta-adrenoceptor was associated with increase in plasma noradrenaline levels during the earlier stages of hypertension. But there is no correlation between beta-adrenoceptor concentrations and plasma noradrenaline levels in the chronic stages of hypertension. No significant difference was found in activities of 5' nucleotidase, which is a marker enzyme of cell membrane, in membrane fractions between the hypertensive hearts and the controls, suggesting that the cardiac hypertrophy is not a determinant factor for change in beta-adrenoceptor. The observed decrease in beta-adrenoceptor concentration may reflect an increase in sympathetic nerve activity during development of hypertension. In the chronic stages of hypertension, additional factors may be involved in the restoration of beta-adrenoceptors. PMID- 6271998 TI - A possible role of the brain angiotensin II receptor binding in development of hypertension. AB - To clarify a role of brain angiotensin II receptor binding in development of hypertension, the regional distribution and extent of specific angiotensin II binding were studied in salt-loaded normotensive (NTR) and spontaneously hypertensive (SHR) rats, and Goldblatt one-kidney hypertensive rat (GB). Further, angiotensin-converting enzyme activity was measured in these rats' brains. In control rats, angiotensin Ii receptor binding was consistently lower in the thalamus, hypothalamus, midbrain, striatum and cortex of SHR rats than in NTR rats. In GB rats, the binding capacity in the thalamus was greater than that of NTR rats. Sodium intake resulted in a rise in the receptor binding capacity in the hypothalamus, thalamus and striatum of SHR rat, whereas it did in a fall in the binding capacity in the hypothalamus, thalamus, striatum, midbrain and cortex of NTR rats. Angiotensin-converting enzyme activity was significantly elevated in the midbrain of salt-loaded SHR rats. PMID- 6271999 TI - Plasma level of norepinephrine and cyclic nucleotides following acute myocardial infarction. AB - The plasma concentrations of norepinephrine (NE), adenosine cyclic 3', 5' monophosphate (cyclic AMP), and guanosine cyclic 3', 5'-monophosphate (cyclic GMP) were measured serially for 2 weeks after the onset of symptoms in 17 patients with acute myocardial infarction (AMI). The mean concentrations of NE in patients without complications were significantly elevated during the first 2 days following AMI. There was a significant correlation between the maximum concentration of plasma NE and of plasma CK. The mean concentrations of plasma cyclic AMP and cyclic GMP in patients without complications were significantly elevated on the first day and for 8 days respectively following AMI. The concentration of plasma cyclic AMP on admission in patients with complications was significantly higher than that in those without complications. There were significant correlations between the maximum concentration of plasma cyclic AMP and those of plasma CK, GOT, and LDH. Significant but weak correlations between the concentration of plasma NE and those of cyclic AMP and cyclic GMP were found. The results of the present study suggest an enhanced sympathetic nervous system activity at an early stage of AMI, a prolonged enhancement of parasympathetic nervous system activity in the course of AMI, and the potential value of plasma cyclic AMP concentrations as a useful index to estimate the seriousness and size of AMI. PMID- 6272000 TI - Effects of captopril and prostaglandin I2 on vasoconstrictor responses to norepinephrine and potassium ions in rat mesenteric artery. AB - In the perfused rat mesenteric vascular bed, the effects of captopril and prostaglandin I2 (PGI2) on the vasoconstrictor responses to norepinephrine or potassium chloride were studied. Captopril or PGI2 in the perfusate attenuated the vascular responses to norepinephrine in a dose-related manner, while these substances had no effect on the vascular contractions induced by potassium chloride. In preparations treated with indomethacin, the inhibitory effect of captopril on the vascular response to norepinephrine was similar to that found in the untreated preparations. These results suggest that, although the effects of captopril on the vascular reactivity is similar to that of PGI2, the direct vascular action of captopril is not mediated by the synthesis of prostaglandins in the vascular bed. PMID- 6272001 TI - Effect of nitroglycerin on the free radical formation of myocardial mitochondria impaired by ethanol. Studies by electron spin resonance (ESR) spectrometry. AB - In order to investigate the direct metabolic effect of nitroglycerin on myocardium, its effect on mitochondrial free radical formation was studied in vitro, using the dog heart mitochondria impaired by ethanol. Free radical concentrations in state 4 respiration or the ratio of free radical concentrations in state 4 to state 1 was used as an index of mitochondrial function. Free radical concentrations were represented as ESR intensity. In normal control, ESR intensity increased from 0.115 +/- 0.030 (mean +/- SD) per mg protein in state 1 to 0.178 +/- 0.017 in state 4 (p less than 0.001). When mitochondria were treated with 10(-4) M ethanol, on the other hand, ESR intensity decreased from 0.163 +/- 0.34 in state 1 to 0.137 +/- 0.019 in state 4 (p less than 0.05). When mitochondria were treated with 10(-6) M nitroglycerin, in spite of the presence of 10(-4) M ethanol, ESR intensity increased from 0.124 +/- 0.23 in state 1 to 0.153 +/- 0.024 in state 4 (p less than 0.05). ESR intensity in state 1 was increased by the treatment of ethanol (p less than 0.05), whereas ESR intensity in state 4 was decreased (p less than 0.001), and these effects were prevented by nitroglycerin (p less than 0.05). The ratio of ESR intensity in state 4 to state 1 demonstrated more remarkably the effects of ethanol and nitroglycerin. Ethanol reduced the ratio from 1.65 +/- 0.47 in the control to 0.86 +/- 0.13 (p less than 0.001) and the reduction was prevented by nitroglycerin to 1.25 +/- 0.23 (p less than 0.01). It is concluded that nitroglycerin prevented the impairment of free radical formation of the mitochondria induced by ethanol, presumably by a mitochondrial membrane-stabilizing action. PMID- 6272002 TI - Effects of sulfinpyrazone, aspirin and propranolol on the isoproterenol-induced myocardial necrosis. AB - Prophylactic effects of sulfinpyrazone (100 mg/Kg), aspirin (5 mg/Kg, 50 mg/Kg), and propranolol (2 mg/Kg, 10 mg/Kg) on myocardial necrosis and hypertrophy induced by isoproterenol were examined. Drugs were administered to rats daily by gavage for a period of 2 weeks and after that isoproterenol (40 mg/Kg) was injected subcutaneously. The control group received gavage of water and isoproterenol injection. The "no infarct" group received gavage of water and saline injection. Premedication of sulfinpyrazone and propranolol significantly preserved myocardial CK activity and decreased cardiac hypertrophy compared with control group (24 hours after isoproterenol injection) while aspirin did not have such effects. Myocardial cyclic AMP concentration significantly increased 30 min after isoproterenol injection in control and all the premedicated rats compared with the "no infarct" rats. The increment of cyclic AMP was not suppressed by sulfinpyrazone and propranolol during this period. Plasma levels of prostaglandins were significantly suppressed by the administration of sulfinpyrazone and 50 mg/Kg of aspirin, and were not suppressed by 5 mg/Kg of aspirin. It was concluded that premedication of sulfinpyrazone and propranolol reduce cardiac necrosis and hypertrophy induced by isoproterenol, but aspirin did not have such cardioprotective effects. PMID- 6272003 TI - Hypotensive response to angiotensin II analogue and angiotensin I converting enzyme inhibitor in pseudo-Bartter's syndrome. AB - We studied the effect of angiotensin II analogue (AII-A) and angiotensin I converting enzyme inhibitor (SQ 14,225) on blood pressure and the renin angiotensin-aldosterone system in a patient with pseudo-Bartter's syndrome, who was a 26-year-old unmarried Japanese woman taking furosemide surreptitiously. The intravenous infusion of AII-A decreased blood pressure from 85/35 to 68/28 mmHg. This decrease in blood pressure was associated with an increment of plasma renin activity (PRA) and a decrement of plasma aldosterone concentration (PAC). Similarly, SQ 14,225 given orally decreased blood pressure to the same extent. An increment of PRA and a decrement of PAC were also observed. These results suggest that the renin-angiotensin system plays a considerable role in maintaining blood pressure in pseudo-Bartter's syndrome. Again, attention has to be paid to the possibility of surreptitious use of diuretics in an adult patient with persistent hypokalemic alkalosis, hyperactivity of the renin-angiotensin-aldosterone system and angiotensin II insensitivity simulating "true" Bartter's syndrome. PMID- 6272004 TI - Granular cell myoblastoma of the bronchus in a child: a case report. AB - Granular cell myoblastoma of of the right main bronchus in a 5-year-old girl is reported. The tumor extended to the lower trachea from the right main bronchus which was occluded completely by the tumor. The diagnosis was established histologically by a obtaining bronchoscopic biopsy specimen. Right pneumonectomy was carried out to control pulmonary suppuration caused by the tumor obstruction of the bronchus. External radiotherapy and interstitial brachytherapy were successful to control the growth of the residual tumor. She has been free from symptoms for about 12 years after the treatment. PMID- 6272005 TI - Inhibitory mechanisms of insulin secretion associated with hypothermic open-heart surgery. AB - In order to clarify the inhibitory mechanism of insulin secretion associated with open-heart surgery, the influence of insulin antagonistic hormones on insulin secretion was studied in 20 patients with congenital heart diseases undergoing open-heart surgery, under simple deep hypothermia. Despite a hyperglycemia, plasma immunoreactive insulin and C-peptide showed no change during the cooling period, while with the exception of plasma human growth hormone, dopamine-beta hydroxylase, immunoreactive glucagon, cortisol and cyclic AMP in plasma, either showed no change, or a decrease during the cooling period. It is assumed that catecholamine, glucocorticoid and glucagon do not play an important role in the inhibitory mechanism of insulin secretion during hypothermic open-heart surgery, and a transient hypofunction of the pancreas as well as the liver and the adrenal gland is probably involved. PMID- 6272006 TI - A non-resectable hepatoma after hepatic artery ligation combined with infusion chemotherapy--an eight-year survival. AB - A 69-year-old man with a large hepatoma of the right lobe extending in part to the medial segment of the left lobe was treated by ligation of the right hepatic artery combined with continuous infusion of a total dose of 5500 mg og 5 fluorouracil (5-FU) into the left hepatic artery. Postoperative celiac arteriogram and liver scintigram revealed a marked regression of the tumor size. After four weeks he was discharged in a satisfactory condition. Seven years and two months after the initial treatment he was readmitted because of a concomitant cancer of the cardia. Total gastrectomy was performed with a favorable result. A second look for the previous hepatic tumor during total gastrectomy revealed a fibrous tissue containing scattered nodules of hepatocellular carcinoma on the right lateral surface of the liver instead of the previous large tumor in the right hepatic lobe. He survived for eight ears after the first operation. PMID- 6272007 TI - Aftercare instruction: an overview. PMID- 6272008 TI - Aftercare instruction: epistaxis. PMID- 6272009 TI - Aftercare instruction: a comprehensive approach. PMID- 6272010 TI - Aftercare instruction: ankle sprains. PMID- 6272011 TI - Aftercare instruction: vaginal bleeding during pregnancy. PMID- 6272012 TI - Nitrosamine-induced transplantable pancreatic ductal adenocarcinoma in inbred Syrian hamsters. AB - A pancreatic ductal carcinoma, designated CBP, was established as a transplantable tumor line in the CB/SsLak inbred strain of Syrian golden hamsters. The tumor, a primary one induced by chronic administration of the nitrosamine N-nitro-bis(2-hydroxyproyl) amine, is a well-differentiated adenocarcinoma that can be propagated by transplantation in syngeneic hamsters. It grows poorly in other hamster strains. The CBP tumor develops in a predictable quantitative manner and metastasizes to regional lymph nodes. Excision of primary transplanted tumor nodules leads to immunity against subsequent secondary tumor challenges. The CBP tumor appears to be a suitable model for biologic and immunologic studies of pancreatic carcinoma in the syrian hamster. PMID- 6272013 TI - Immunoprecipitation of intracisternal A-particle-associated antigens from preimplantation mouse embryos. AB - Murine intracisternal A-particles (IAP) are endogenous retrovirus particles expressed in large numbers in certain neoplastic tissues and during early stages of normal preimplantation mouse (outbred ICR) embryogenesis. IAP-associated antigens synthesized by preimplantation mouse embryos were precipitated by a rabbit (New Zealand White) antibody prepared against IAP from murine myeloma 104E. Characterization of the embryo immunoprecipitates with the use of one- and two- dimensional polyacrylamide gel electrophoresis revealed that a group of five proteins was synthesized by two- to eight-cell stages, but only three of these (molecular weight of 67,000, 69,000, and 73,000) correlated with the morphologic expression of IAP in embryos. ONly the proteins with molecular weights of 75,000 and 77,000 were detected in morulae and blastocysts, when embryos were IAP negative. This is the first biochemical identification of endogenous retrovirus associated antigens in preimplantation mouse embryos. PMID- 6272014 TI - [Medical topics: hormone receptor abnormalities]. PMID- 6272015 TI - Characterization of acidification by the isolated perfused rat kidney: evidence for adaptation by the distal nephron to a high bicarbonate diet. AB - To characterize acidification by the distal nephron of the intact kidney independent of alterations in buffer availability, we subjected isolated rat kidneys perfused with glucose as the sole substrate to stepwise acidification of the perfusate. In response to progressive perfusate acidification with hydrochloric acid, a maximal pH gradient between urine and perfusate, which averaged 1.71 +/- 0.12, was achieved at a mean perfusate pH of 6.89 +/- 0.04. The maximum pH gradient was increased when sulfuric acid rather than hydrochloric acid was used to acidify the perfusate, and it was decreased by 0.67 when amiloride (10(-5) M) was added to the perfusate. Thus, hydrogen ion transport by the distal nephron of the intact rat kidney appears to be amenable to study, and it responds similarly to the hydrogen ion pump of anuran urinary epithelia. Kidneys from animals subjected to a variety of dietary regimens were studied in a response to stepwise perfusate acidification with hydrochloric acid. Ammonium excretion averaged 0.49 +/- .03 mumoles/min and did not differ significantly between any of the dietary groups. Chronic acidosis and the ingestion of either a low or high salt diet had no influence on the maximal pH gradient. Neither a low nor a high potassium diet affected the pH gradient, suggesting that the difference in urine pH between these two conditions in vivo is the result of differences in ammonia production. Ingestion of a high bicarbonate diet significantly decreased the pH gradient to 1.20 +/- 0.09. Thus, an adaptive change in distal nephron hydrogen ion transport occurs in the rat kidney in response to chronic ingestion of alkali. PMID- 6272016 TI - [Timolol response in denervational syndromes (author's transl)]. AB - In eyes with pupillotony (Argyll Robertson) the effect of locally applied timolol maleate (0.5%) on intraocular pressure is considerably less pronounced than in normal control eyes. In Fuchs' heterochromic cyclitis the timolol response is the same as in controls. Infrared thermography of the anterior segment of the eye showed that the maximum drop in IOP was preceded by a fall in temperature. These findings suggest that the IOP-lowering effect of timolol depends on parasympathetic innervation and that timolol decreases the blood flow in the ciliary body. PMID- 6272017 TI - [Alterations of te ocular fundus in a patient with acute lymphoblastic leukemia (author's transl)]. AB - A report on a boy with acute lymphoblastic leukemia, who was treated with cytostatic drugs and radiation. In the course of treatment retinal changes occurred in one eye; at first they seemed to indicate a solitary ocular relapse. Later on characteristic serological findings made it appear likely that the child was suffering from cytomegalic retinitis. The differential diagnosis of leukemic ophthalmopathy and the prognosis of leukemia with ophthalmopathy are discussed. PMID- 6272018 TI - Iatrogenic multiorgan silicone inclusions in dialysis patients. PMID- 6272019 TI - SEM observation of the stromal surface of the monkey submandibular gland treated with HCl-collagenase. PMID- 6272020 TI - Effects of colchicine and cytochalasin B on vasopressin- and cyclic adenosine monophosphate-induced changes in toad urinary bladder. AB - Coincident with an increase in water permeability, the ridge-like surface structures of toad bladder granular cells transform to individual microvilli after stimulation with vasopressin (VP) or cyclic adenosine monophosphate (cAMP) by a mechanism that is yet to be defined. To explore the possible role of microtubules and microfilaments in this cell response, colchicine and cytochalasin B were employed to determine whether interference with the function of these components of the cytoskeletal system would prevent the VP- and cAMP induced conversion of ridges to microvilli. Incubation of toad urinary bladders in 10(-4) M colchicine for 4 hours or 10(-5) M cytochalasin B for 90 minutes before stimulation with 20 mU. per ml. of VP markedly inhibited osmotic water flow. However, neither agent prevented the striking conversion of ridges to surface microvilli induced by VP and cAMP as seen with scanning electron microscopy. In addition, the ridges characteristic of granular cells were maintained in control bladders incubated with colchicine or cytochalasin B, but left unstimulated. Under the conditions of these experiments, these findings suggest that microtubules and microfilaments are not essential for maintenance of normal surface configuration in granular cells of toad urinary bladder, and that they are not involved in the mechanism responsible for VP- and cAMP-induced surface changes that occur in association with increased water permeability of this epithelium. PMID- 6272021 TI - Effect of cholera toxin on secretion of mucin by explants of guinea pig trachea. AB - Cholera toxin (CT) elicits a dosage-dependent increase in mucin secretion by explants of guinea pig trachea. Concomitantly, the mucin in goblet cells of the mucosa and submucosal glands is depleted. This effect is realized in the absence of cell injury, as assessed morphologically and by the assay of culture medium for the release of acid phosphatase. Mucosal concentrations of cyclic AMP increase after exposure to CT. However, the stimulatory effects on secretion appear to be independent of the cyclic nucleotide, as exogenous dibutyryl cyclic AMP and cyclic GMP fail to increase secretion, and theophylline, a phosphodiesterase inhibitor, also is ineffective. The stimulatory effect of CT is decreased by preincubation of the explants with inhibitors of microtubules (nocodazole) and microfilaments (cytochalasin D) in a dosage-dependent manner. Addition of the calcium chelator, ethylene glycol bis(beta-aminoethyl ether)-N,N' tetraacetic acid, with CT also inhibits the secretory response. CT appears to stimulate mucin secretion by tracheal epithelial cells by a mechanism independent of cyclic nucleotide activation but requires intact microtubules, microfilaments, and exogenous calcium ions. PMID- 6272022 TI - Mineralocorticoid activity of 21-deoxyaldosterone derivatives: structure-function studies. PMID- 6272023 TI - Immunologic inhibition of estrogen binding and action in preputial-gland cells and their subcellular fractions. PMID- 6272024 TI - Gestational changes in hamster adrenocortical function. PMID- 6272025 TI - The efficiency of different coupling procedures for the linkage of oestriol-16 alpha-glucuronide, oestrone-3-glucuronide and pregnanediol-3 alpha-glucuronide to four different enzymes. PMID- 6272026 TI - Combined effects of angiotensin II and ACTH on cortisol production: in vitro studies on isolated guinea-pig adrenal cells. PMID- 6272027 TI - Conjugated and unconjugated oestrogens in men with hyperoestrogenism and in normal men: the day-to-day variation and the relationship between the different oestrogens. PMID- 6272029 TI - The safe-neighborhood hypothesis of junk DNA. PMID- 6272028 TI - Sarcomas of the breast: homogenous or heterogenous? AB - Eighteen cases of sarcomas of the breast are reported. Cases could be divided into two groups histogenetically. Group A, malignant cystosarcoma phylloides (14 cases); and Group B, pure sarcomas (4 cases). The malignant cystosarcoma phylloides occurred in the forth decade, had linger duration of illness invariably followed by sudden increase in the size of the tumor, had a striking mucoid and slimy look grossly, and showed discernible epithelial elements histologically. In the event of their recurrence the ductal elements were not seen microscopically. Contrarily, Group B cases presented in the sixth decade were rapidly growing from the beginning, and grossly were fleshy and haemorrhagic. Microscopically these did not reveal any epithelial component. There was poor correlation between cytologic malignancy and the biologic behavior in Group A. Mode of treatment seems more important in determining the subsequent course of the disease. Simple mastectomy with follow-up appears to be adequate in the majority of cases. PMID- 6272030 TI - Frequency encoded biochemical regulation is more accurate than amplitude dependent control. PMID- 6272031 TI - Intracellular perfusion. PMID- 6272032 TI - Solid-phase immunoassay methods for quantitation of IgG and viral antibodies in cerebrospinal fluid and its electrophoretic fractions. PMID- 6272033 TI - Early biochemical reactions in the lung of sheep exposed to asbestos: evidence for cyclic AMP accumulation in bronchoalveolar lavage fluids. PMID- 6272034 TI - Hepatobiliary cancer--surgical considerations. AB - Most cancers of the hepatobiliary system will have grown beyond the limits of curative resection by the time they become clinically evident. This reality has fostered therapeutic nihilism, and most physicians and surgeons in their pessimism have failed to study the early modes of spread of these tumors- patterns of growth that are relevant to proper treatment of those patients who do have lesions that can be removed with hope of achieving a cure. Moreover, anatomic and technical complexities may beget surgical reluctance in the management of potentially curable lesions. Therefore, this review is offered for orientation and perspective for those who would hope to offer optimal treatment for patients who have primary cancers of the liver, gallbladder, or biliary ducts. The review includes considerations of (1) surgical anatomy, (2) modes of spread, (3) assessment of resectability, (4) surgical technique, and (5) results of operative treatment in relation to "curative" resection. Also offered are some guides to palliative surgical management of tumors that have reached the hepatic hilus. PMID- 6272035 TI - [Acute pancreatitis disclosing metastasis of bronchial cancer]. PMID- 6272036 TI - [Usefulness of various cellular systems for replication and isolation of enteroviruses]. PMID- 6272037 TI - [Tests for the presence of bacteriophages in live virus vaccine]. PMID- 6272038 TI - Inactivation of cyclic GMP-dependent protein kinase by N alpha-tosyl-L-lysine chloromethylketone. PMID- 6272039 TI - Dextroamphetamine infusions in normals result in correlated increases of plasma beta-endorphin and cortisol immunoreactivity. PMID- 6272040 TI - Surgical stress in humans is accompanied by an increase in plasma beta-endorphin immunoreactivity. PMID- 6272041 TI - Comparative metabolic clearance rates of beta-endorphin and beta-lipotropin in humans. PMID- 6272042 TI - Comparison of analgesic and body temperature responses to intrathecal beta endorphin and D-Ala2-D-Leu5-enkephalin. PMID- 6272043 TI - Intracerebroventricular beta-endorphin increases food intake of rats. PMID- 6272044 TI - Interaction of GnRH agonists, pituitary hormones and corticosteroids on progesterone secretion in the male rat. PMID- 6272045 TI - Immunoreactive alpha-melanotropin and beta-endorphin in the toad pars intermedia: dissociation in storage, secretion and subcellular localization. PMID- 6272047 TI - High affinity beta-2-adrenergic receptors in mononuclear leucocytes: similar density in young and old normal subjects. PMID- 6272046 TI - Biological inactivation of enkephalins and the role of enkephalin-dipeptidyl carboxypeptidase ("enkephalinase") as neuropeptidase. PMID- 6272048 TI - Effects of beta-endorphin fragments on plasma levels of vasopressin. PMID- 6272049 TI - Opiates and enkephalins: a common binding site mediates their analgesic actions in rats. PMID- 6272050 TI - The action of angiotensin on the isolated perfused cat heart. PMID- 6272051 TI - Plasma ACTH and cortisol levels in depressed patients: relation to dexamethasone suppression test. PMID- 6272052 TI - Does endogenous cyclic GMP inhibit potassium stimulated 45Ca uptake by P2 fraction from rat brain? PMID- 6272053 TI - [Acupuncture: hypnosis or placebo effect?]. PMID- 6272054 TI - [Wilms' tumor in adults. Case report and follow up clinico-pathologic examinations of 96 literature cases]. PMID- 6272055 TI - [Cytomegalic retinitis in immunosuppressed patients]. PMID- 6272056 TI - Methods for the preparation of enzyme-antibody conjugates for use in enzyme immunoassay. PMID- 6272057 TI - Ubiquinones: stereochemistry and biological implications. AB - Proton NMR and 13C-NMR studies on the configuration of CoQn homologues show that the polyisoprenoid side-chain is in the all-trans configuration and confers a higher degree of rigidity to the quinones with respect to the acyl-chains of the phospholipids within the membrane bilayer. The quinonoid ring appears to be specifically involved in the redox function of the coenzyme while the side-chain length only affects the lipophilicity of the molecule. LIS data show that the ring strongly interacts with metals as a consequence of the high pi-electron density on the carbonyls that is somewhat larger on the carbonyl oxygen to alpha to the isoprenoid chain. PMID- 6272058 TI - Characterization of the electrogenic sodium channel from rat brain membranes using neurotoxin-dependent 22Na uptake. AB - The sodium channel was studied in osmotically-sensitive membrane preparations from rat brain and in innervated and chronically denervated rat soleus and extensor digitorum longus muscles. These experiments were undertaken in order to define a set of parameters for sodium channel function at the subcellular level to be used as a measure of retention of channel integrity upon subsequent isolation of the channel. Various neurotoxins and drugs were employed to control the permeability of the brain membranes to 22Na and the sodium-conductance properties of the muscles. Batrachotoxin (ED50 = 0.2 micrometer), veratridine (ED50 = 1 micrometer), or grayanotoxin I (ED50 = 30 micrometers) stimulated 22Na uptake in brain membranes is inhibited in an apparently uncompetitive manner by the sodium channel blocking agents tetrodotoxin and saxitoxin in a simple competitive manner by Ca2+ and in a partial or allosteric competitive manner by lidocaine and procaine. This 22Na uptake assay, which can be equated to a measure of equilibrium toxin binding, shows dependence on the concentration of the membranes and is sensitive to pH, temperature, ionic strength, and the ionic composition of the media. Parallel biophysical studies on sodium channels in rat muscle show that the properties of the sodium channel are similarly affected by these agents. PMID- 6272059 TI - Translocation of sequences encoding antibiotic resistance from the chromosome to a receptor plasmid in Salmonella ordonez. AB - Salmonella ordonez strain BM2000 carries kanamycin (Km), ampicillin (Ap), spectinomycin (Sp), chloramphenicol (Cm), tetracycline (Tc), and sulfonamide (Su) resistance and production of colicin Ib (Cib). The Km and Cib characters were carried by a 97 kb IncI1 plasmid (pIP565). In addition to the Km and Cib traits, all or part of the other antibiotic resistance (R) determinants could be transferred by conjugation from S. ordonez to Escherichia coli where all the acquired characters are borne by an IncI1 plasmid, designated complete or partial composite plasmid respectively. DNA from pIP565 and composite plasmids and total DNA from strain BM2000 were studied by agarose and polyacrylamide gel electrophoresis following digestion with restriction endonucleases, and by Southern hybridization. These comparative analyses enabled us a) to show that acquisition by pIP565 of resistance to all or some of the antibiotics was due to the insertion of a single DNA fragment into the receptor plasmid; b) to detect two types of composite plasmids with regard to the specificity of insertion into pIP565 and the mapping of the inserts; c) to demonstrate that the ApCmSpSuTc resistance determinants were integrated into S. ordonez BM2000 chromosomal DNA; d) to map the restriction fragments of the translocatable sequence integrated into strain BM2000 chromosome or into pIP565. The results obtained suggest that two distinct mechanisms for the translocation of the R determinants coexist in S. ordonez BM2000. Recombination between two of the four directly repeated copies of the IS-like sequence (IS1522) present in S. ordonez chromosome leads to the circularisation of all or part of the ApCmSpSuTc R determinants and is followed by either 1) a second recombination with the copy of IS1522 in pIP565 (Type I composite plasmids), or 2) transposition of precise groups of characters in various sites of pIP565 (Type II composite plasmids). PMID- 6272060 TI - Identification of the E. coli groNB(nusB) gene product. AB - The E. coli groNB(nusB) gene product has been previously shown to be necessary for bacteriophage lambda N protein function. The product of the groNB gene has been identified on SDS polyacrylamide gels after infection of UV-irradiated E. coli cells with various lambda groNB+ transducing phage derivatives. It is a polypeptide with an apparent molecular weight of 14,000 daltons. Transducing phage carrying either a deletion or an amber mutation in the groNB gene fail to synthesize the 14,000-Mr polypeptide chain upon infection of a sup+ host. However, am+ revertants of the lambda groNBam phage do induce the synthesis of the polypeptide. PMID- 6272061 TI - Plasmid pGB301, a new multiple resistance streptococcal cloning vehicle and its use in cloning of a gentamicin/kanamycin resistance determinant. AB - Streptococcal plasmid pGB301 is an in vivo rear ranged plasmid with interesting properties and potential for the molecular cloning of genes in streptococci. Transformation of S. sanguis (Challis) with the group B streptococcal plasmid pIP501 (29.7 kb) gave rise to the deletion derivative pGB301 (9.8 kb, copy number 10) which retained the multiple resistance phenotype of its ancestor (inducible MLS-resistance, chloramphenicol resistance). Among the eight restriction endonucleases used to physically map pGB301 were four that cleaved the plasmid at single sites yielding either sticky (HpaII, KpnI) or blunt-ends (HpaI, HaeIII/BspRI). Passenger DNA derived from larger streptococcal plasmids (pSF351C61, 69.5 kb; pIP800, 71 kb) was successfully inserted into the HpaII site and, by blunt-end cloning into the HaeIII/BspRI site. The gentamicin/kanamycin resistance gene of pIP800 was expressed by recombinant plasmids carrying the insert in either orientation. Insertion of passenger DNA into the HaeIII/BspRI site (but not the HpaII site) caused instability of adjacent pGB301 sequences which were frequently deleted, thereby removing the chloramphenicol resistance phenotype. The vector pGB301 has a remarkable capacity for passenger DNA (inserts up to 7 kb) and the property of instability and loss of a resistance phenotype following insertion of passenger DNA into the HaeIII/BspRI site should facilitate the identification of cloned segments of DNA when using this plasmid in molecular cloning experiments. PMID- 6272062 TI - Construction of hybrid plasmids containing the lysA gene of Escherichia coli: studies of expression in Escherichia coli and Saccharomyces cerevisiae. AB - The lysA gene of Escherichia coli has been cloned from a lambda transducing phage on various plasmids, present in different copy numbers in bacterial cells. Synthesis of the product of this gene, diaminopimelate (DAP)-decarboxylase, and its regulation have been studied. Expression does not follow a simple gene dosage effect, maximal expression already being obtained with a six-copy plasmid. This result suggests that either a positive or an autogenous regulatory mechanism is involved. We also used one of the hybrid plasmids to look for expression of the bacterial lysA gene in Saccharomyces cerevisiae. The results indicate that the product of the E. coli gene is not actively translated in yeast. PMID- 6272063 TI - Genetical and structural analysis of a group of lambda ilv and lambda rho transducing phages. AB - Eight lambda ilv C transducing phages generated from E. coli K12 secondary site lysogens have been analysed genetically and physically. Two of them carry, in addition, the rho gene and its promotor region, but not the cya gene. The ilv O 603 mutation has been located between ilv G and ilv E. Electrophoretic analysis of the proteins synthesized by these phages in a system of UV irradiated cells allowed us to assign molecular weights of 55000 and 66000 daltons to the ilv C and the ilv D gene products, respectively, and to show that an ilv G-encoded polypeptide of 60000 daltons is made from an ilv O- but not from an ilv O+ phage. The expression of the ilv G gene is discussed in the light of the recent finding of a promoter-attenuator region lying upstream to ilv G. Finally, we have found that one of the lambda ilv phages does not have the classical structure of a transducing phage. PMID- 6272064 TI - Isolation and characterization of cAMP suppressor mutants of Escherichia coli K12. AB - We have isolated spontaneous and chemically induced revertants of cya mutant strains of Escherichia coli. Three different classes of revertants were obtained. One class consisted of primary site revertants; a second class was pseudorevertants that had phenotypically reverted to wild type but retaining the original cya mutant and the third class of revertants, designated csm, were pseudorevertants hypersensitive to exogenous cAMP. Transductional analysis of the csm mutation indicated the mechanism of suppression in these strains was intergenic. The csm mutation and hypersensitivity to cAMP map in or near the crp gene. Growth of the csm strains on PTS (phosphoenolpyruvate phosphotransferase system) and non-PTS substrates was inhibited by 5 mM cAMP. The csm strains were found to accumulate toxic levels of methylglyoxal when grown on non-PTS substrates in the presence of exogenous cAMP. All csm strains were sensitive to catabolite repression mediated by alpha-methylglucoside. Revertants selected as resistant to cAMP fell into four major classes that could be distinguished by their fermentation patterns in the presence and absence of cAMP as well as by their growth response to streptomycin in the presence of cAMP. PMID- 6272065 TI - Identification of the nusB gene product of Escherichia coli. AB - Escherichia coli nusB mutants fail to support the activity of a phage lambda gene product, pN, which regulates phage gene expression by influencing transcription termination. We report the identification of the nusB protein on SDS polyacrylamide gels as a 14,500 dalton protein. PMID- 6272066 TI - The number of ribosomal RNA genes in Mycoplasma capricolum. AB - We have examined the number of rRNA genes in Mycoplasma capricolum (KID) by hybridization of Bg/II-, EcoRI- and Xbal-digests of DNA to [3'-32P] 16S, 23S and 5S rRNAs according to the Southern procedure (1975). All the restriction gels gave two radioactive bands with three kinds of rRNA. Furthermore, band positions were indistinguishable from one another when 16S, 23S and 5S rRNAs were used as probes, indicating that each band contains sequences corresponding to the 3' termini of 16S, 23S and 5S rRNAs. It is thus concluded that Mycoplasma capricolum chromosome carries at least two sets of genes for 16S, 23S and 5S rRNAs. PMID- 6272067 TI - Bacillus subtilis bacteriophages SP beta c1 is a deletion mutant of SP beta. AB - The restriction fragment patterns of two mutants forms of the temperate Bacillus subtilis bacteriophage SP beta have been examined. The DNA of a heat-inducible mutant, SP beta c2, which has a molecular size of 128 kilobases (kb), yields the same restriction pattern as the wild type SP beta c+ DNA. The DNA of a clear plaque mutant, SP beta c1, has a molecular size of 117 kb, and is deleted for an 11 kb region of phage DNA. Neither SP beta c1 nor SP beta c2 DNA is cleaved by the endonuclease HaeIII. PMID- 6272068 TI - pED100, a conjugative F plasmid derivative without insertion sequences. AB - The largest HindIII fragment of F includes the entire replication and transfer regions, and its circularisation with ligase gave the conjugative plasmid pED100. This plasmid, which contains none of the F insertion sequences, was essentially unable to mobilise the E. coli chromosome or to give integrative suppression of a dnaA strain. PMID- 6272069 TI - Antimicrobial activity of some alkyl esters of gallic acid (3,4,5, trihydroxybenzoic acid) against Escherichia coli NCTC 5933 with particular reference to n-propyl gallate. AB - The growth inhibitory and bactericidal activities of eight alkyl esters of gallic acid towards Escherichia coli NCTC 5933 have been determined. A previously suggested role for gallic acid and its esters as shikimate antimetabolites could not be substantiated. No induction of gross changes in cell morphology was observed. Bactericidal activity was accompanied only be very slight leakage of general ionic materials from the bacteria. Propyl gallate did not appear to uncouple oxidative phosphorylation from respiration as indicated by its failure to stimulate proton translocation across the cytoplasmic membrane. PMID- 6272070 TI - Limitations of erythromycin in whooping cough. PMID- 6272071 TI - Aetiology of viral gastroenteritis: a review. AB - Over the past seven to eight years, several virus groups have been shown to be associated with gastroenteritis. They are adenoviruses, astroviruses, caliciviruses, coronaviruses. Norwalk-like viruses and rotaviruses. In infants and young children, rotaviruses are the single most important aetiological agents of acute gastroenteritis in terms of numbers of cases and also of patients requiring admission to hospital. In older children and adults, the aetiology is less clear, but the present state of our knowledge indicates that Norwalk-like viruses are one of the major causes. Most of these gastroenteritis-producing viruses cannot be propagated satisfactorily in laboratory cell-culture systems, and electron microscopy is the chief method of detection. Immune electron microscopy is used to demonstrate specific antibody increases during infection and to show antigenic differences between morphologically similar viruses. These techniques are relatively insensitive, and the development of a cell-culture system for the propagation of these "gastroenteritis viruses" would greatly facilitate epidemiological studies and vaccine development. PMID- 6272072 TI - Fybogel in haemorrhoid treatment. PMID- 6272073 TI - Campylobacter as a cause of acute enteritis in children in South Australia. I. A 12-month study with controls. AB - During a 12-month period, we tested faecal samples from 386 children with acute enteritis and 332 controls by light and electron microscopy, and by bacterial and viral culture for pathogens, especially to assess the importance of campylobacter. Campylobacter alone was responsible for the illness in 17 patients (5%), and was second to salmonella among the bacterial agents, which were predominant in summer. Overall, rotavirus was the commonest identifiable cause of acute enteritis and was especially important in winter (with a rate of 49% in August). Thirty-five patients showed two or more agents. In 117 patients (30%) no pathogen was isolated or identified, suggesting that there are as yet unidentified agents in acute enteritis. PMID- 6272074 TI - Campylobacter as a cause of acute enteritis in children in South Australia. II. Clinical comparison with salmonella, rotavirus and non-specific enteritis. AB - The clinical features of 17 children with campylobacter enteritis were compared with 17 age- and sex-matched children with enteritis due to salmonella, rotavirus or those in whom there was no identifiable pathogen. Prominent clinical features of campylobacter enteritis included fever, diarrhoea, vomiting, blood in stools and periumbilical pain. Dehydration was uncommon, compared to rotavirus and non specific enteritis. The acute illness was self-limited, in spite of prolonged asymptomatic faecal excretion of the organism. This prolonged carriage increases the risk of cross infection. No patient with campylobacter required antibiotic therapy. Recurrent epidoses of diarrhoea were seen in three children but on no occasion was campylobacter the cause. This study has demonstrated a marked similarity between campylobacter and salmonella enteritis, making clinical distinction virtually impossible. Bloody diarrhoea, a feature of bacterial infections, was absent in rotavirus and non-specific enteritis. PMID- 6272075 TI - [Prevalence of functional bronchial obstruction and identification of groups at risk in a population of industrial workers]. PMID- 6272076 TI - Analysis of the transformation of human lymphocytes by Epstein-Barr virus III. Induction of early events by P3HR-1 strain without subsequent immortalization. AB - T-cell depleted human cord blood lymphocytes were exposed to P3HR-1 strain of Epstein-Barr virus (EBV) and simultaneous observations of immunofluorescence, cellular morphology, and autoradiography were carried out in individual cells. Soon after infection, nuclear antigen (EBNA) synthesis, blastogenesis, and DNA synthesis occurred, as was previously observed B95-8 strain EBV infection. Although mitosis followed with characteristic cell aggregate formation, the cell proliferation was temporary and death followed in about 2 weeks. The synthesis of the early antigens (EA) and the viral capsid antigen (VCA) were not significant. These findings seem to indicate that the strain P3HR-1 EBV is capable of inducing early events of transformation in primary human B-lymphocytes, but the cells infected in this way have a short life span. PMID- 6272077 TI - Cystosarcoma phylloides: a case presentation. AB - A case of cystosarcoma phylloides is presented here showing the natural history of this disease both clinically and pathologically from 1962 through 1979, and demonstrating the management of this disease by surgery and by chemotherapy with the use of adriamycin at 25 mg/m2 daily, times three days, every 28 days, which to out knowledge has not been used in this particular setting. PMID- 6272078 TI - Myocardial injury immediately following adriamycin administration. AB - A patient is described who developed chest pain during adriamycin administration given for the first time for small cell carcinoma of the lung. This was associated with the acute onset of atrial fibrillation with a rapid ventricular response. Although no electrocardiographic changes of myocardial ischemia developed, serum isoenzyme studies suggested that myocardial injury had occurred. The patient recovered but died suddenly three weeks later. Physicians should be aware of possible life-threatening arrhythmias developing during or shortly after adriamycin administration. PMID- 6272079 TI - [Trends in studies of the pathogenesis and clinical picture of parasitic diseases in the 11th 5-year plan]. PMID- 6272082 TI - Solubilization of a [3H]cimetidine binding site from rat brains. A clonidine sensitive H-2 receptor subtype? PMID- 6272084 TI - Two different modifications of the neuroblastoma X glioma hybrid opiate receptors induced by N-ethylmaleimide. PMID- 6272083 TI - Demonstration of [3H]cyclazocine binding to multiple opiate receptor sites. PMID- 6272081 TI - Adenosine 3',5'-phosphate in fungi. PMID- 6272085 TI - Opiate receptors in the rat brain. Specific labeling of multiple membrane components with [3H]etorphine? PMID- 6272080 TI - Plasmids, drug resistance, and gene transfer in the genus Streptococcus. PMID- 6272086 TI - Scatchard analysis of opiate receptor binding. PMID- 6272087 TI - Influences of temperature, detergents, and enzymes on glutamate receptor binding and its regulation by calcium in rat hippocampal membranes. PMID- 6272088 TI - Ethanol enhances [3H]diazepam binding at the benzodiazepine-gamma-aminobutyric acid receptor-ionophore complex. PMID- 6272089 TI - Regulation of cyclic AMP metabolism in human platelets. Sequential activation of adenylate cyclase and cyclic AMP phosphodiesterase by prostaglandins. PMID- 6272090 TI - Catecholamine-induced desensitization of adenylate cyclase in rat glioma C6 cells. Evidence for a specific uncoupling of beta-adrenergic receptors from a functional regulatory component of adenylate cyclase. PMID- 6272092 TI - The ionic channel of the acetylcholine receptor. Regulation by sites outside and inside the cell membrane which are sensitive to quaternary ligands. PMID- 6272091 TI - Evidence for alpha-adrenergic activation and inactivation of phosphorylase in hamster adipocytes. PMID- 6272093 TI - Inhibition of voltage-sensitive sodium channels in neuroblastoma cells by antiarrhythmic drugs. PMID- 6272094 TI - Quinone-stimulated superoxide formation by subcellular fractions, isolated hepatocytes, and other cells. PMID- 6272095 TI - Nucleoside analogues with clinical potential in antivirus chemotherapy. The effect of several thymidine and 2'-deoxycytidine analogue 5'-triphosphates on purified human (alpha, beta) and herpes simplex virus (types 1, 2) DNA polymerases. PMID- 6272096 TI - Hormone-sensitive adenylate cyclase. Mutant phenotype with normally regulated beta-adrenergic receptors uncoupled with catalytic adenylate cyclase. PMID- 6272097 TI - [Physical mapping of Streptomyces coelicolor A3(2). V. Structural modifications of actinophage phiC43 DNA molecules]. AB - As shown by genetical and physical methods, the total preparation of phiC43 phage obtained after spontaneous induction of the prophage from S. lividans 803 strain is a heterogenous population. The wild-type phage (phi C43 wt) is only represented in 5--10% of the population. The majority of phage variants are not able to establish the lysogenic state. The structure of DNA molecules of some phages from the total preparations was characterized by electron microscopy of DNA heteroduplexes. Molecules of phiC43 wt DNA appeared to be completely homologous to those of recently studied phiC62 phage, except for two small regions of approximately 0.3 kb in the central part. Phage variants defective in establishment of the lysogenic state were distributed to two groups. One of them consists of deletion variants, the other--deletion/insertion variants. Deletions in DNA molecule of all nonlysogenizing phage overlap. The region of overlapping seems to be responsible for establishment of the lysogenic state. In the same region, deletion of DNA molecules of mutant phiC311 yg2 has been located. Three deletion/insertion variants contain homologous foreign sequences of various length. It is likely that these insertions are fragments of the host chromosomal DNA. PMID- 6272098 TI - [Structure containing cellular mRNA in picornavirus infections]. AB - The fate of cellular mRNA upon infection of Krebs-2 ascites carcinoma cells with encephalomyocarditis (EMC) virus was investigated. The cell mRNA was discovered in a structure with a sedimentation coefficient of about 100S and a buoyant density of 1.50--1.519 g/cm3 during active virus-specific synthesis (3.0--4.0 hr post infection). The template activity of the 100S structure in a cell-free protein-synthesizing system and of mRNA isolated from it was studied and the nature of synthesized products was analyzed. It was shown that the 100S structure seems to be translationally inactive. On the contrary, the RNA isolated from its is functionally active. PMID- 6272099 TI - [Protection of segments of replicative form I of phiX174 phage DNA recognized by HindII, BspRI, and AluI restrictases by Escherichia coli RNA-polymerase]. AB - The preincubation of a DNA with E. coli RNA polymerase provides its partial protection against the HindII, BspRI and AluI cleavage giving possibility to determine the location of RNA polymerase tight-binding sites. Using this approach about 16 RNA polymerase tight-binding sites were detected on replicative form of phiX174 phage DNA. The protection degree of each of these sites depended on the preincubation conditions. Some of the protected sites hit the known phiX174 promoters and rho-dependent terminators and the other were distributed along the whole phiX174 DNA molecule. Many of them could be considered as potential promoters because they contain all the necessary elements specifying the real promoter sequences. At least some of the intrinsic promoter elements could be observed next to the rest of protected sites. One of the protected sites (R6b/l) is located in phiX174 DNA region which is very similar to the cAMP-CRP-controlled promoter sequences. It was confirmed that phiX174 DNA has two B promoters positioned by Sanger on the phiX174 nucleotide map, according to our data obtained by RNA polymerase protection experiments along with RNA product analysis of the R8 DNA fragment transcription in vitro. PMID- 6272100 TI - [Ribosomal genes of the loach Misgurnus fossilis L. Isolation and restriction analysis]. AB - The ribosomal DNA of the teleost fish--loach has been isolated from sperm DNA by CsCl density gradient centrifugation. The rDNA sediments on density gradients by two heavy satellites beta = 1.715 and greater than 1.720. The DNA of the first satellite (1.715) was separated and treated by restrictases EcoRI and BamHI. It was shown that there are two EcoRI-sites in rDNA of loach, locating in 18S and 28S rRNA coding sequences. From tandem of repeating ribosomal genes EcoRI cuts out the fragment with homogeneous length-3 megadaltons (constant fragment) and heterogeneous population of fragments 11-13 megadaltons (major) and 7-8 megadaltons (minor fraction). The constant fragment contains mostly 28S coding sequence, and the heterogeneous fragment--18S coding sequence. The data indicate that the ribosomal genes of the loach as well as other higher eucaryotes were organized in genome as tandem of repeating units with heterogeneous length (10-16 megadaltons, 14.5-24 kb) due to heterogeneity of the length of nontranscribed spacer. PMID- 6272101 TI - Amyloidosis and lung cancer. A morphological and histochemical study. AB - A study performed on a sample of 120 subjects who died of lung carcinoma and were examined at autopsy in order to verify the presence of amyloid substance particularly localized in the tumoral tissue, is reported. In the positive cases (4 oat cell carcinomas and 1 squamous cell carcinoma), the histochemical nature of depositions was investigated to assess their belonging to the primitive or secondary type, and to the APUD or immune type. The doubtful results induced the authors to suppose an analogous behaviour of oat cell carcinoma and squamous cell carcinoma. The explanation of this phenomenon is still unclear and it should be looked for starting from two hypotheses: an analogy between oat cell carcinoma and squamous cell carcinoma both belonging to the APUD system, or simply a non specific phenomenon (intratumoral amyloid deposition). PMID- 6272102 TI - Histochemical and histoenzymatic liver changes in Guerin tumour-grafted rats. AB - In order to follow up the morphological changes of the rat liver as a reaction to the tumour graft, three groups of Guerin tumour-grafted animals were used: a first control group, a second one treated with leucotrophine (LT) and a third one treated with LT and thiamine diphosphate (TDP). The tumour-grafted rats showed hepatic changes affecting mainly the organelles involved in cellular respiration and synthesis, as well as some morphological changes of Kupffer cells expressing an increased endocytosis. The protection by immunostimuli determined the diminution of histochemical and histoenzymatic changes both in hepatocytes and Kupffer cells. PMID- 6272103 TI - Cytomegalovirus inclusion body endocervicitis: significance of CMV inclusions in endocervical biopsies. PMID- 6272104 TI - The hyaline eccrinoma. PMID- 6272105 TI - [Sick by passive smoking?]. PMID- 6272106 TI - Modification of MR mutator activity in repair-deficient strains of Drosophila melanogaster. AB - In drosophila, MR (male recombination) second chromosomes are known to act as mutators and recombination-inducers in males. One explanation of MR- induced effects is that MR causes breaks at specific sites where, subsequently, insertion sequences become integrated. To examine the extent to which excision and incorporation of DNA sequences by MR is affected by enzymatic pathways involved in DNA repair, the following experiments were carried out. MR-chromosomes were introduced into males deficient for excision (mei-9a) or post-replication repair (mei-41D5) or into males carrying both repair-deficient mutations. MR activity was recorded by the occurrence of visible mutations at the sn (singed) and ras (raspberry) loci. The spontaneous mutation frequency for sn is 0.2 . 10-5 (1/490 000) and for ras, 0.4 . 10-5 (2/490 000). When MR is introduced into repair proficient males the frequency of sn mutations is 51 . 10-5 (16/30 795) and of ras mutations is 19. 10-5 (6/30 795). In MR males deficient for both excision and post-replication repair (mei-9a mei-41D5) mutator activity is significantly enhanced; the frequency of sn mutations amounts to 225 . 10-5 (174/77 219) and of ras mutations to 135 . 10-5 (104/77 219). mei-9a or mei-41D5 alone also leads to an enhancement of the mutation frequencies, but this effect is of borderline significance. PMID- 6272107 TI - A calcium regulated adenosine triphosphatase in Entamoeba histolytica. AB - Axenically grown trophozoites of Entamoeba histolytica (NIH-200 strain) contain an active ATPase (170 nmol PO4/min per mg protein) with maximal activity at pH 8.8, a high affinity for ATP (Km approximately 40 micro M) and an absolute and specific requirement for Ca2+. The activation by Ca2+ shows positive cooperativity (nH = 2.48) at calcium concentrations below 8 micro M and no cooperativity between 8 and 25 micro M. The latter concentration fully saturates the enzyme. The observed activity is insensitive to oligomycin, ouabain and ruthenium red and is unaffected by a range of inhibitors of electron transport and uncouplers of oxidative phosphorylation. The enzyme exhibits structure bound latency and is tightly bound to cellular membranes. It is sedimentable ( greater than 80%) by high speed centrifugation of cell homogenates which are either protected osmotically or in which subcellular structures are damaged by sonication or treatment with Triton X-100. Arrhenius plots of V in the temperature range of 0-38 degrees C are linear without breaks, similar to other pyrophosphatases of E. histolytica. The calculated activation energy is 14.8 kcal/mol. This finding as well as the failure of phospholipase treatment to affect activity indicate that interactions with membrane lipids play no role in the catalytic function of this tightly membrane-bound ATPase. PMID- 6272108 TI - Susceptibility of Entamoeba histolytica to oxygen intermediates. AB - To explore the susceptibility of the extracellular protozoan, Entamoeba histolytica, to toxic oxygen intermediates, trophozoites were exposed to fluxes of O2, H2O2, and OH. generated enzymatically by the glucose oxidase and xanthine oxidase reactions. HM-1 trophozoites were resistant to O2, but were readily killed by H2O2 alone. OH. and 1O2 were not required for effective amebicidal activity. The addition of peroxidase and halide enhanced trophozoite killing by H2O2. Sonicates of amebae contained virtually no catalase and little glutathione peroxidase activity which may contribute to susceptibility to H2O2. Coupled with our previous studies with Toxoplasma gondii and Leishmania spp. these observations indicate that there is a broad spectrum of susceptibility of intra- and extracellular pathogenic protozoa to killing by oxygen intermediates. PMID- 6272109 TI - Pneumocystis carinii pneumonia and mucosal candidiasis in previously healthy homosexual men: evidence of a new acquired cellular immunodeficiency. AB - Four previously healthy homosexual men contracted Pneumocystis carinii pneumonia, extensive mucosal candidiasis, and multiple viral infections. In three of the patients these infections followed prolonged fevers of unknown origin. In all four cytomegalovirus was recovered from secretions. Kaposi's sarcoma developed in one patient eight months after he presented with esophageal candidiasis. All patients were anergic and lymphopenic; they had no lymphocyte proliferative responses to soluble antigens, and their responses to phytohemagglutinin were markedly reduced. Monoclonal-antibody analysis of peripheral-blood T-cell subpopulations revealed virtual elimination of the Leu-3 / helper/inducer subset, an increased percentage of the Leu-2 + suppressor/cytoxic subset, and an increased percentage of cells bearing the thymocyte-associated antigen T10. The inversion of the T/ helper to suppressor/cytotoxic ratio suggested that cytomegalovirus infection was an important factor in the pathogenesis of the immunodeficient state. A high level of exposure of male homosexuals to cytomegalovirus-infected secretions may account for the occurrence of this immune deficiency. PMID- 6272110 TI - Severe acquired immunodeficiency in male homosexuals, manifested by chronic perianal ulcerative herpes simplex lesions. AB - Four homosexual men presented with gradually enlarging perianal ulcers, from which herpes simplex virus was cultured. Each patient had a prolonged course characterized by eight loss, fever, and evidence of infection by other opportunistic microorganisms including cytomegalovirus, Pneumocystis carinii, and Candida albicans. Three patients died; Kaposi's sarcoma developed in the fourth. All were found to have depressed cell-mediated immunity, as evidenced by skin anergy, lymphopenia, and poor or absent responses to plant lectins and antigens in vitro. Natural-killer-cell activity directed against target cells infected with herpes simplex virus was depressed in all patients. The absence of a history of recurrent infections or of histologic evidence of lymphoproliferative or other neoplastic diseases suggests that the immune defects were acquired. PMID- 6272111 TI - Hypoglycemia associated with non-islet-cell tumor and insulin-like growth factors. PMID- 6272112 TI - Opportunistic infections and Kaposi's sarcoma in homosexual men. PMID- 6272113 TI - Cytomegalovirus glomerulopathy in renal allografts. PMID- 6272114 TI - Long-term follow-up of persons inadvertently inoculated with SV40 as neonates. PMID- 6272115 TI - Surgery of soft tissue sarcomas in children. AB - Surgery of soft tissue sarcomas in children has been modified by the introduction of multidisciplinary treatment so that major amputation and exenterations are now rare in the management of these tumors in children. Surgery must still be well planned for total removal of the tumor with preservation of limbs and, in most instances, the function of pelvic organs. The commonest soft tissue sarcomas in children are embryonal rhabdomyosarcoma (RMS), fibrosarcoma, and synovial sarcoma. Treatment of embryonal RMS can now achieve a 2-year survival of 80% at all sites. Surgery and irradiation are used to control the primary tumor and multidrug chemotherapy to control metastases or prevent dissemination of localized tumor. Fibrosarcoma in children, usually a low-grade, extra-abdominal desmoid lesion, does not respond to radiation treatment or chemotherapy, and management is by surgery alone. It shows a marked tendency to local recurrence, and multiple local resections may be necessary for cure. Ninety percent of the children with this tumor can be salvaged by surgery and careful follow-up. The management of synovial sarcoma is surgical and similar to that of RMS. Although not radiosensitive as is RMS, this tumor has responded well to multidisciplinary treatment. Adequate resection is now followed by a chemotherapy protocol similar to that used in osteogenic sarcoma. The smaller numbers of these tumors and their varied natural history make evaluation of treatment difficult. Other soft tissue sarcomas seen with extreme rarity in children are liposarcoma, angiosarcoma, and neurosarcoma. These tumors are treated with the same protocol as that of RMS. PMID- 6272117 TI - Soft tissue sarcomas of childhood: the differential diagnostic dilemma of the small blue cell. AB - In its histologic features, embryonal rhabdomyosarcoma (RMS), the prototype of malignant soft tissue tumors in childhood, summarizes the problems associated with the diagnosis of this entire group of neoplasms. Many of the tumors that do not fulfill the criteria for RMS have been designated "sarcomas of uncertain histogenesis." The introduction of the concept of a soft tissue equivalent of Ewing's sarcoma may have eased the semantic anxiety without improving our conceptual understanding. It is thought that the embryonal RMS, Ewing's sarcoma, and other are derived from a primitive mesenchymal cell. Another separate category of "small blue cell tumors" are those which presumably originate from the primitive neuroepithelium. Some of the diagnostic terms applied to this category are "neuroepithelioma," "medulloepithelioma," and "peripheral neuroblastoma." Because most of these tumors are hormonally inactive and electron microscopy is not performed, the diagnosis is infrequently considered or proved. The recently described small cell tumor of thoracopulmonary origin is likely a malignant neuroepithelial neoplasm. Hematopoietic tumors, such as non-Hodgkin's malignant lymphomas, granulocytic sarcoma, and malignant histiocytosis, may appear in the soft tissues as the initial manifestation of these system diseases. A final group of malignant soft tissue tumors are the fibrohistiocytic ones with a biphasic pattern of small round cells and spindle cells. It now has become increasingly difficult for the pathologist to satisfy his clinical colleagues with the diagnosis of "undifferentiated malignant tumor" in a child. PMID- 6272116 TI - Sarcomas of bone in childhood: pathologic aspects. AB - Some sarcomas of bone are not characteristically childhood tumors. Within the context of osteosarcomas, telangiectatic and parosteal (juxtacortical) variants deserve recognition. Among chondrosarcomas, which ordinarily are not tumors that occur in children, the mesenchymal variant should be recognized in children especially, this lesion may be extraosseous. Fibrous histiocytoma and its malignant variant have recently emerged as morphologically specific lesions, occasionally osseous, among other spindle cell tumors. Ewing's sarcoma, which is predominantly of osseous origin, consists of undifferentiated fragile cells. The elaborate spectrum of topographic, cytologic, nuclear, and other morphologic features that fall within the designation Ewing's sarcoma are described. PMID- 6272118 TI - A new era in mammalian gene mapping: somatic cell genetics and recombinant DNA methodologies. AB - Mammalian gene mapping techniques are now sufficiently advanced to contribute significantly to prenatal diagnosis and to human molecular genetics. Restriction fragment mapping can be used to place polymorphic genetic markers at random sites within the genome, and these sites used to assign genes responsible for disease conditions to a chromosomal region. Somatic cell genetic techniques can then be applied to saturate that region with additional restriction fragment markers, some of which will be closely linked to the disease gene. Closely linked restriction fragment markers, especially flanking pairs of markers, can act as predictors for the transmission of defective genes to offspring. A series of tightly linked flanking restriction markers might in addition contribute to the eventual isolation and cloning of the disease gene itself. PMID- 6272119 TI - Structure of the pro alpha 2 (I) collagen gene. AB - Fifty-four kilobase pairs (kbp) of cloned chicken DNA containing the entire 38 kbp pro alpha 2 (I) collagen gene have been isolated and characterized. DNA sequence analysis of a select 4 kbp of the gene has precisely described 14 exons which comprise one-third of the sequences encoding the triple-helical domain of the collagen protein. These exons range in size from 45 to 108 base pairs (bp), are all multiples of the 9 bp that code for the repeating triplet, Gly-X-Y, and have an average size of 70 bp. About 50 introns interrupt this gene. Nevertheless, introns do not separate the coding sequences for the ends of the central triple-helical structural domain and the ends of the propeptide domains. PMID- 6272120 TI - Anti-transferrin receptor monoclonal antibody and toxin-antibody conjugates affect growth of human tumour cells. PMID- 6272121 TI - Point mutation in the TATA box curtails expression of sea urchin H2A histone gene in vivo. PMID- 6272122 TI - E. coli ribosomal protein L10 inhibits translation of L10 and L7/L12 mRNAs by acting at a single site. PMID- 6272123 TI - Glucocorticoids regulate expression of dihydrofolate reductase cDNA in mouse mammary tumour virus chimaeric plasmids. AB - Fusions between the mouse mammary tumour virus long terminal repeat and a mouse dihydrofolate reductase cDNA have been constructed in a SV40 vector. When these plasmids are transferred into recipient cells, the production of dihydrofolate reductase is regulated by glucocorticoid hormones. These results define a hormonally responsive region of the viral genome. PMID- 6272124 TI - Sodium and potassium channels in demyelinated and remyelinated mammalian nerve. PMID- 6272125 TI - Isolation of a new type C retrovirus (HTLV) in primary uncultured cells of a patient with Sezary T-cell leukaemia. PMID- 6272126 TI - Antibodies in human sera reactive against an internal structural protein of human T-cell lymphoma virus. PMID- 6272127 TI - The v-sis transforming gene of simian sarcoma virus is a new onc gene of primate origin. PMID- 6272128 TI - Structure of the glycoprotein gene in rabies virus. PMID- 6272129 TI - Streptozotocin and alloxan induce DNA strand breaks and poly(ADP-ribose) synthetase in pancreatic islets. PMID- 6272130 TI - Collagenase and hyaluronidase pretreatment induces sensitivity to d-tubocurarine in frog sciatic nerve. PMID- 6272131 TI - Studies on the role of Na+, K+ and Cl- ion permeabilities in K+-induced release of 3H-noradrenaline from rat brain slices and synaptosomes and in its presynaptic alpha-adrenergic modulation. PMID- 6272132 TI - The effect of isoprenaline on plasma concentrations of immunoreactive beta endorphin and beta-lipotropin in the conscious rat. AB - The effect of the beta-adrenoceptor agonist isoprenaline on the plasma concentrations of beta-endorphin (beta-E) and beta-lipotropin (beta-LPH) was investigated in conscious rats. Isoprenaline (i.m.) elevated plasma beta endorphin-like immunoreactivity (beta-EI) as measured by radioimmunoassay of unextracted plasma, with peak values 24 min after drug administration. This effect was dose-dependent. The lowest effective dose of isoprenaline was 15 micrograms kg-1; 240 micrograms kg-1 exerted a maximum effect, raising plasma beta-EI about ten-fold above control values. Plasma vasopressin concentrations also increased in response to isoprenaline following a time-course identical to that of plasma beta-EI. (+/-)-Propranolol (1 mg kg-1) but not phentolamine (10 mg kg-1) rendered isoprenaline (240 micrograms kg-1) injections almost ineffective. Because of the cross-reactivity of beta-LPH in the radioimmunoassay used, plasma was extracted by means of a cation exchange resin and subjected to gel chromatography on a Sephadex G-50 column, avoiding artefactual degradation of the peptides. In isoprenaline-treated rats about 50% of the beta-EI behaved similar to human beta-LPH, whereas 45% co-migrated with human beta-E; immunoreactivity corresponding to beta-LPH or beta-E comprised about 70% or 30%, respectively, in the plasma extract of vehicle-treated rats. Dexamethasone pretreatment reduced the isoprenaline-induced increase in plasma beta-EI by 87%, but left the simultaneous elevation of plasma vasopressin concentrations unchanged. These data demonstrate that isoprenaline stimulates beta-LPH and beta-E release in vivo. The possibility of an interrelationship between vasopressin and beta-E release is discussed. PMID- 6272133 TI - Loperamide reduces the intestinal secretion but not the mucosal cAMP accumulation induced by choleratoxin. AB - The effect of loperamide on net fluid transport and epithelial cAMP accumulation was tested in choleratoxin-exposed ligated colon loops of the rat in vivo. Purified choleratoxin (50 micrograms/ml saline, for 5 h) induced net secretion and doubled cAMP levels in comparison with saline-treated controls. Loperamide (4 mg/kg intragastrically) reduced this secretion by 75%, without diminishing cAMP accumulation; it had no effect on basal fluid transport or cAMP. The data suggest that the opiate analogue interferes with the secretory process at a point beyond the cAMP increase caused by activation of adenylate cyclase. PMID- 6272134 TI - [Properties of the slow excitatory postsynaptic potential in mammalian sympathetic ganglia neurons]. AB - Slow EPSPs evoked in the neurons of the rabbit isolated superior cervical ganglion were studied using intracellular microelectrodes. Two types of EPSPs occurring in different neurons were found. The type I slow EPSPs showed an increase during hyperpolarization of the membrane and a decrease during its depolarization. Input resistance of the neurons during the response either decreased or remained unchanged. The type II slow EPSPs were increased by depolarization and decreased by hyperpolarization with the reversal potential 78.9 +/- 3.6 mV. Depolarization evoked by acetylcholine or carbocholine was followed by an increase in the input resistance in 53% of neurons with reversal potential -83.2 +/- 6.7 mV. It is concluded that in the first group of the neurons the nature of the slow EPSP is similar to that of ordinary EPSP. The main component underlying the ionic mechanism of slow EPSP in the other group of the neurons is a decrease in potassium conductance of the membrane. PMID- 6272135 TI - [Stimulus-dependent blockade of node of Ranvier sodium channels by ethmozine]. AB - The effect of antiarrhythmic drug ethmozine on sodium channels in Ranvier node was studied by the voltage clamp technique. Both outside and inside application of ethmozine induced a decrease of sodium current I Na, the time course of I Na and the sodium inactivation being unchanged. The ethmozine-induced Na channel blockade induced tonic (stationary) and phasic (transient stimulus-dependent) components. The tonic blockade of I Na developed slowly and could be accelerated by frequent electric stimulation of the membrane. The phase dependent blockade became more profound with an increase in the pulse rate or amplitude of depolarizing pulses. The prolonged (1 s) membrane depolarization did not bring about an additional blockade of I Na. It is concluded that the phasic blockade is due to the interaction of ethmozine with open Na channels. The noninactivating batrachotoxin-modified Na channels were insensitive to ethmozine. It is found that the increase in outside potassium concentration from 2.5 to 20mM induced both a decrease of the tonic blockade and an increase of the phasic one. The possible nature of the ionic blockade is discussed. The effect of ethmozine is compared with that of tertiary and quaternary local anesthetics. PMID- 6272136 TI - [Sodium pump reconstruction in lipid vesicles]. AB - Reconstitution of purified lubrol-solubilized preparation of Na,K-ATPase from the calf brain was performed in liposomes and transport characteristics of this system were investigated. The system was demonstrated to preserve vectoral features of the sodium pump. Addition of ATP to the medium evoked active influx of sodium and efflux of rubidium in proteoliposomes. Ouabain inhibited transport processes when enclosed into proteoliposomes, i.e. from the side opposite to the ATP-sensitive one. PMID- 6272137 TI - [Effect of magnesium ions on presynaptic inhibition of monosynaptic reflexes]. AB - In experiments on spinal narcotized cats perfusion of lumbosacral spinal cord through central canal with artificial cerebrospinal fluid containing high concentration (20-46 mM) of magnesium ions led to reversible depression of negative DRP as well as to depression of prolonged "presynaptic" inhibition of extensor monosynaptic reflexes produced by repetitive impulse volleys in group I flexor muscle afferents. Magnesium did not cause a depression of monosynaptic reflex discharges in spinal ventral roots. PMID- 6272138 TI - [Factors responsible for generation of multiple discharges in motor neurons of the nucleus of the facial nerve in cats]. AB - Peculiarities of multiple discharges of facial nucleus motoneurons evoked to stimulation of caudal trigeminal nucleus were studied in acute experiments on cats by intracellular recording technique. The multiple discharge is shown to appear on the basis of gradual depolarization of the motoneuron membrane resulted from effective summation of high-frequency excitatory synaptic influences from the caudal trigeminal nucleus. Factors supporting the development of the gradual depolarization are dendritic location of synaptic terminals of trigeminal projecting neurons, dendritic origin of depolarizing after-potentials and high input resistance of the facial motoneuron membrane. PMID- 6272139 TI - [Prostaglandins, their role in the body and relation to the central nervous system]. PMID- 6272140 TI - [Survival period of patients with gliomata or glioblastomata under anti convulsive therapy (author's transl)]. PMID- 6272141 TI - [How can we give schizophrenic patients better treatment?--An integration of new pathogenic and therapeutic concepts (author's transl)]. PMID- 6272142 TI - Angiotensin I-converting enzyme in the diagnosis and management of sarcoidosis. PMID- 6272143 TI - [Evidence, in uremic man, of the role of beta 2 adrenergic receptors in the secretion of parathyroid hormone and calcitonin (author's transl)]. AB - In order to determine the effect of beta-blocking agents on secretions of parathyroid hormone and calcitonin, 9 patients with renal failure were given single doses of propranolol (a blocker of the beta 1 and beta 2 receptors) or an equivalent amount of metoprolol (a beta 1 selective agent). Propranolol causes a decrease of plasma parathyroid hormone (p less than 0.02) as well as of calcitonin (p less than 0.05) whereas metoprolol has no effect on the plasma levels of these hormones. These findings suggest that parathyroid tissue and thyroid C cells have receptors that are exclusively of the beta 2 type which are modulating the secretion of parathyroid hormone and calcitonin. PMID- 6272144 TI - Benzodiazepine receptor binding in young, mature and senescent rat brain and kidney. AB - Clinical reports have described age-altered pharmacological effects of anxiolytic drugs especially an increased susceptibility to their sedative actions. In order to test whether such changes may be due to age-related alterations in central benzodiazepine receptors, 3H-flunitrazepam binding was assayed in the frontal cortex and cerebellum of young, mature and senescent rats. The numbers of 3H flunitrazepam binding sites and their affinity was determined by Scatchard analysis of saturation isotherms and the relative abundance of type I and type II benzodiazepine receptors was assessed by drug-inhibition studies using diazepam and the triazolopyridazine, CL 218,872. In addition, age related changes in the kidney and hippocampus of the Ro5-4864-sensitive benzodiazepine receptor were studied using 3H-Ro5-4864. No age-related alterations were noted in the binding characteristics of 3H-flunitrazepam. Furthermore, drug-inhibition of 3H flunitrazepam binding by diazepam and CL 218,872 was nearly identical in young, mature and senescent rats, indicating that also the ratio of type I and type II receptors does not change with age. Binding of 3H-Ro5-4864 to membranes from rat hippocampus was not age-related. However, a significant decrease in 3H-Ro5-4864 binding to kidney membranes was demonstrated. Hence, central benzodiazepine receptors appear unaltered in the senescent rat model of aging. The clinical findings of an increased susceptibility to the sedative effects of benzodiazepines in the elderly may therefore be attributed to pharmacokinetic variables, or to events occurring secondarily to receptor activation. PMID- 6272145 TI - Changes in noradrenergic neurotransmission in rat cerebellum during aging. AB - This study compared the electrophysiological effects of locally applied cyclic adenosine-mono-phosphate (N6cAMP) between Purkinje neurons from young (4-month) and old rats (15-months and older). Purkinje neurons from young rats were significantly more sensitive to locally applied norepinephrine and N6cAMP than neurons from old rats. GABA sensitivity between the two groups was unaltered. Our results suggest that the locus of the adrenergic subsensitivity observed in older animals may reside to a large extent at or beyond the level of cAMP generation. PMID- 6272146 TI - Conversion and binding of tetraiodothyronine in developing rat brain. AB - In this study we have examined whether rat brain nuclear thyroid hormone receptors bind T4 or metabolites of T4 and whether there is a developmental change in brain T4 metabolism and binding. Developing animals were injected with trace [125I]3',5'-tetraiodothyronine ([125I]T4) and after sacrifice brain nuclear and cytoplasmic fractions were examined to determine whether their radioactivity was represented by the infected [125I]T4 or any of its metabolites. Of the radiothyronines specifically bound to the nucleus, 90% was found to be triiodothyronine ([125I]T3) and 10% was [125I]T4. Of the cytoplasmic, protamine sulfate-precipitable fraction, 40% was [125I]T4 and 60% [125I]T3. Inasmuch as the percentage of [125I] T3 found in plasma during the same postinjection interval was similar to that present as contaminant of the injected material, it was concluded that brain [125I] T3 derives from local monodeiodination of T4 to T3. The main developmental change observed was a marked decline in the total cytoplasmic and nuclear [125I] T4 uptake. However, with development, the T3/T4 ratio remained constant in the nuclear fraction while it decreased in the cytoplasmic fraction. It is concluded that although T3, deriving from monodeiodination of T4, is the main form of thyroid hormone that regulates brain development by its binding to brain nuclear receptors, the fact that T4 is the most available form during the critical period makes it, indirectly, very important to brain development. Further, the decline observed with development in T4 uptake and monodeiodination to T3, may contribute to the concomitantly declining role of thyroid hormones on brain tissue. PMID- 6272147 TI - The development of rat dorsal root ganglia in three experimental media containing Landry-Guillain-Barre, alcoholic peripheral neuropathy or normal adult sera. AB - Development of rat dorsal root ganglia in vitro mimics the development of rat dorsal root ganglia in vivo when grown in medium containing human cord serum. When serum from patients either in the acute phase of Landry-Guillain-Barre syndrome or with alcoholic peripheral neuropathy is substituted for human cord serum, maturation is inhibited. Development of fetal relationships occurs between the Schwann cells and axons and many axons become enclosed within the processes of a single Schwann cell. Formation of myelin does not occur. The substitution of normal adult blood for cord serum also inhibits maturation, but unlike development with neuropathic serum, the Schwann cells and axons do not become associated and myelination does not occur. PMID- 6272148 TI - Spongiform polioencephalomyelopathy caused by a murine retrovirus. II. Ultrastructural localization of virus replication and spongiform changes in the central nervous system. AB - The development of murine retrovirus induced spongiform polioencephalomyelopathy was studied sequentially by electron microscopy. During the initial 30 days, viral infection of the central nervous system, as evidenced by viral budding from membranes, was limited to the endothelial cells and pericytes. Viral particles were observed in the lumen of blood vessels, extracellular spaces and astrocytic endfeet surrounding blood vessels, but no morphological evidence of productive infection was found in astrocytes or neurons during early development of vacuolation. The earliest lesions in the neuropil consisted of swelling of astroglia followed by vacuolation, initially in axons and dendrites and later in neuronal and astrocytic soma, where vacuoles appeared to arise from dilated cisternae of the Golgi apparatus. Vacuoles contained only amorphous debris and fragments of membranes. Virions budding aberrantly into vacuoles were seen only in mice surviving beyond 35 days. Numerous reactive astrocytes were observed, but inflammatory cells were absent. The ultrastructural changes were remarkably similar to those described in scrapie, Kuru, and Creutzfeldt-Jakob disease. PMID- 6272149 TI - Evidence for a possible dopaminergic control of pituitary alpha-MSH during ontogenesis in mice. AB - This study was aimed at determining the alpha-MSH and ACTH contents of the neurointermediate lobe (NIL) of the mouse hypophysis during ontogenesis, as well as the ability of the gland, incubated in a perfusion system, to respond to dopamine (DA) and high potassium (K+). We showed that: (1) the NIL content of alpha-MSH exhibited a biphasic pattern of evolution, characterized by a dramatic increase appearing between postnatal days 3 and 5. By contrast, NIL ACTH content followed a completely different pattern of evolution; (2) both DA and high K+ reversibly inhibited MSH release from superfused NIL, the latter effect being more pronounced with the use of pituitaries from 7-day-old than from 1-day-old mice; (3) the inhibitory influence of high K+ was impaired by haloperidol pretreatment. From these data, as well as from morphological observations, it appears that functional DA receptors seem to be present on melanotrophs of the developing hypophysis in mice and that the increase in alpha-MSH in IL cells observed shortly after birth may result from the onset of inhibitory influence exerted by dopaminergic innervation on hormonal secretion. This view, however, does not preclude possible effects of other hypothalamic releasing and/or inhibiting factors. PMID- 6272150 TI - Alpha-receptor-mediated facilitation of somatosensory cortical neuronal responses to excitatory synaptic inputs and iontophoretically applied acetylcholine. PMID- 6272151 TI - Effects of ACTH on pain responsiveness in mice: interaction with morphine. PMID- 6272152 TI - Biology of gliomas: potential clinical implications of glioma cellular heterogeneity. PMID- 6272153 TI - Absorbable or nonabsorbable suture materials for closure of the dura mater? AB - This study was undertaken to determine which suture material would give the best results for closure of the dura mater. Because there is no need for the suture to remain longer than until healing has occurred and as remaining foreign material will predispose to foreign body reactions and infection, it was considered important to test new absorbable materials like polyglactin 910 (Vicryl) and polyglycolic acid (Dexon) for closure of the dura mater. The reactions from these materials were compared to those of silk and polyester (Ethibond) in dogs 60 days after the suture of dural incisions. The quality of healing with respect to the smoothness of the subdural surface, the presence of adhesions between sutures and the brain surface, the degree of absorption of the material, and reactions around the sutures were evaluated. Vicryl gave the best results, providing a smooth subdural surface without adhesions. It was almost totally absorbed when healing was completed, in contrast to Dexon and the other materials. The cellular reaction around Vicryl was slight. Silk, Ethibond, and Dexon protruded from the subdural surface and adhesions to the arachnoid were common. When the dura mater was removed, these adhesions tore off the leptomeninges with their vascular supply to the cortex. The results of these experiments seem to justify the recommendation of Vicryl for suturing of the dura mater. Such an absorbable material decreases the risk of foreign body reactions, and infections will be more easily combated because of the absence of foreign material. The lack of subdural adhesions is an advantage if reoperation is necessary. PMID- 6272154 TI - Redundant nerve roots of the cauda equina. PMID- 6272155 TI - A study of the rat septohippocampal pathway using anterograde transport of horseradish peroxidase. PMID- 6272156 TI - Projections of the pulvinar-lateral posterior complex to visual cortical areas in the cat. PMID- 6272157 TI - Selective bidirectional transport of [3H]d-aspartate in the pigeon retino-tectal pathway. PMID- 6272158 TI - Reorganization of muscle afferent projections accompanies peripheral nerve regeneration. PMID- 6272159 TI - Development of central control of norepinephrine turnover and release in the rat heart: responses to tyramine, 2-deoxyglucose and hydralazine. PMID- 6272160 TI - A nerve stump dependent appearance of junctional and perijunctional acetylcholine receptors in organ culture. PMID- 6272161 TI - Properties of nerve endings with small granular vesicles in the distal colon and rectum of the guinea-pig. PMID- 6272162 TI - [Macrophage reactions of the cerebrospinal fluid in the course of viral and bacterial meningitis]. PMID- 6272164 TI - [Curie therapy with 192 iridium on a moulded apparatus in the treatment of tumors of the hard palate]. AB - Complete regression of the primary tumour was observed when a shaped apparatus loaded with 192Ir wires was used in the treatment of 6 patients with neoplasia of the hard palate, whereas very slight changes were noted in another subject. Fatal locoregional adenopathy appeared after 6 months in one of these 6 patients without prejudice to the cure obtained in the primary site. PMID- 6272163 TI - Detection of brain tumours in patients with focal epilepsy by regional cerebral blood flow measurements. PMID- 6272165 TI - Origins of the projections of the superior colliculus to the dorsal lateral geniculate nucleus and the pulvinar in the rabbit. AB - The origins of the projections of the superior colliculus to the dorsal lateral geniculate nucleus and to the pulvinar in Dutch-belted rabbits were investigated using horseradish peroxidase (HRP) methods. Following injections of HRP in the dorsal lateral geniculate nucleus, retrogradely labeled neurons were found in the upper two-thirds of the stratum griseum superficiale of the ipsilateral superior colliculus. Most of the labeled somata were spindle-shaped, and their major axes tended to be perpendicular to the surface of the superior colliculus. In contrast, following injections of the pulvinar, labeled neurons were found in the lower third of the ipsilateral stratum griseum superficiale. In these cases, the labeled somata were larger than those labeled following dorsal lateral geniculate injections and were multipolar in shape. PMID- 6272166 TI - Neurones situated outside the isthmo-optic nucleus and projecting to the eye in adult birds. AB - The centrifugal projection to the eye has been studied by retrograde horseradish peroxidase (HRP) transport in adult pigeons and chickens. About 1500 large neurones outside the contralateral isthmo-optic nucleus and 20 or so ipsilateral ectopic cells contain HRP 0.5-3.5 days after intravitreal HRP injections. The number of ectopic neurones which project to the contralateral eye is more than 20% of the number of labelled cells within the isthmo-optic nucleus. In contrast to the monopolar isthmo-optic neurones, the ectopic cells are a distinct population of large multipolar cells. Previously it has been suggested that many of these cells degenerate during the development of the chick. This study shows that they persist in the adult avian brain where they can be detected in large numbers by the presence of retrogradely transported HRP, provided that an HRP chromogen reaction of sufficient sensitivity is used. The exact target for their axons is unknown. PMID- 6272168 TI - Thalamocaudate projection neurons with a branching axon to the cerebral motor cortex. AB - Sixty-nine thalamocaudate projection neurons in ventroanterior (VA: 32 cells), centrolateral (CL: 12 cells) and centrum medianum or parafascicular (CM-Pf: 25 cells) nuclei of the cat were identified by antidromic response to stimulation of the caudate nucleus (CD). Twenty-two VA and 3 CL neurons in this sample were proved to have a branching axon projecting on both CD and the cerebral motor cortex (Mcx). Most of the branching VA neurons showed repetitive discharges synchronous with the cortical recruiting response elicited by repetitive stimulation (6-10/sec) of the pallidum. Such activities of the VA neuron which would induce the motor cortical recruiting response, were proved to be conveyed to CD. PMID- 6272167 TI - Stimulation of resistance to 6-hydroxydopamine in a human neuroblastoma cell line by nerve growth factor. AB - Resistance of human neuroblastoma cell line SK-N-SH-SY5Y (SY5Y) to the neurotoxin 6-hydroxydopamine (6-OHDA) was established by exposure of the cells to nerve growth factor (NGF). SY5Y cells display several properties of immature sympathetic nerve cells, including morphological responses to nerve growth factor and susceptibility to cytolysis by 6-OHDA. High resistance to 6-OHDA was achieved by culturing SY5Y cells with NGF. Protection persisted when NGF was absent for 24 h or when cells were treated with colcemid. In contrast, dibutyryl cyclic AMP did not stimulate resistance to 6-OHDA. NGF-treated cells were also resistant to H2O2, a toxic product of 6-OHDA autoxidation. PMID- 6272169 TI - Primary ovarian hydatidiform mole: addition of a sixth case to the literature. PMID- 6272170 TI - [Role of adrenergic receptors in the etiology of primary glaucoma]. PMID- 6272171 TI - Retinoid-induced growth inhibition of herpesvirus-transformed marmoset lymphoblastoid cell lines. AB - The in vitro proliferation of herpesvirus-transformed marmoset lymphoblastoid cell lines (LCL) was inhibited by retinoic acid and retinyl acetate. Both Herpesvirus-ateles-and Herpesvirus-saimiri-transformed LCL with T-cell characteristics and Epstein-Barr-transformed LCL with B-cell markers were more sensitive to retinoic acid than to retinyl acetate. Inhibition of LCL proliferation was dependent on retinoid concentration and became apparent only after 3-4 days of exposure. In vitro assays for marmoset LCL sensitivity to retinoids may indicate the potential usefulness of these compounds in the chemoprevention of virus-induced lymphoma in vivo. PMID- 6272173 TI - [Care of patients with implanted artificial heart valves]. PMID- 6272172 TI - Enamel in odontodysplasia. AB - Microradiographs of a tooth affected by odontodysplasia showed a clear delineation between the hypoplastic enamel and dentin. Two different zones, one presenting a prismatic structure and the other a globular structure, were found in the enamel layer. In areas where normal prismatic enamel was found, a scalloped dentinoenamel junction and a normal mantle dentin layer were present. Crystallites in those areas were of normal size but the intercrystalline spaces were enlarged. Hypoplastic globular enamel consisted of smaller, closely packed crystallites resembling a type of enameloid found in teleost fishes. Round calcified bodies attached or unattached to the hypoplastic enamel were composed of still smaller irregularly distributed crystallites. It is suggested that the outer hard tissues in odontodysplasia involve different processes. The prismatic enamel zones are formed by the first differentiated ameloblasts. The hypoplastic enamel is due to a decrease in the number of ameloblasts. A localized loss of ameloblasts induces lack of enamel in places. The round calcified bodies attached or unattached to the hypoplastic enamel are most likely formed without any involvement of the ameloblasts. PMID- 6272174 TI - [Neonatal hypocalcemia with hyperparathyroidism in the mother (author's transl)]. AB - A report on two siblings, on formula feeding, who had fits in the second week of life. Cause of the hypocalcemic cramps was a formerly undetected hyperparathyroidism in the mother. PMID- 6272175 TI - [Cytomegalovirus in subacute infant pneumonitis (author's transl)]. AB - A subacute pneumonic disease of a young infant is described. Insidious onset, afebrile course, tachypnea, staccato cough, disseminated crepitations on auscultation, signs of infiltration and hyperexpansion on chest X-ray, eosinophilia as well as elevation of immunoglobulin fractions G and M suggested infection with chlamydia trachomatis. Microbiologic investigations, however, documented active infection with cytomegalovirus. Hence these symptoms were either caused by both organisms or constitute a rare monorganic manifestation of connatal cytomegalovirus disease. The syndrome of subacute pneumonia in the young infant might not necessarily depend on the infection with a specific organism. PMID- 6272176 TI - Increase of beta-endorphin in cerebrospinal fluid after removal of ACTH-secreting pituitary adenoma. AB - beta-Endorphin and methionine(met)-enkephalin in cerebrospinal fluid (CSF) were measured before and after removal of an adrenocorticotropic hormone-(ACTH) secreting adenoma in Cushing's disease by a sensitive radioimmunoassay and a radio-receptor assay, respectively. After tumor resection, the level of ACTH in plasma markedly decreased from 82.6 +/- 22.7 pg/ml to 16.7 +/- 4.1 pg/ml (mean +/ S.E., n = 4). It was found that the level of beta-endorphin in CSF significantly increases from 32.0 +/- 4.5 pg/ml to 61.8 +/- 10.7 pg/ml (P less than 0.05) after tumor resection, while the level of metenkephalin in CSF remained unaltered. This result suggests that hypophysectomy induces an increase of beta-endorphin in CSF. PMID- 6272177 TI - An autopsy study of hepatocellular carcinoma in Hong Kong. AB - Two hundred and eighty-seven autopsy cases of hepatocellular carcinoma (HCC) in Chinese were reviewed. The analyses included histological study of the tumour and of the non-cancerous liver tissue, the cause of death and metastases. Bleeding of oesophageal varices was more frequent but rupture of tumour less common in cases associated with cirrhosis than in those without cirrhosis. There was a significantly higher incidence of bilobar involvement by tumour in the clear cell type of HCC and in cases unassociated with cirrhosis compared to other histological types of HCC and HCC with cirrhosis, possibly because of longer survival of the former groups. A strong association was found between cirrhosis and hepatic fibrosis with HCC and hepatitis B surface antigen (HBsAg), suggesting an oncogenic effect of chronic persistent hepatitis B virus (HBV) infection on hepatocytes. PMID- 6272178 TI - The response of three inbred strains of rat to the carcinogen 1,2 dimethylhydrazine. AB - Rats of 3 inbred strains (DA, HS and AS2) were dosed with 1,2-dimethylhydrazine 2HCl (DMH) or saline by gavage weekly for 10 wk. DA and HS rats showed little overt toxic reaction but all AS2 rats died following DMH doses of 30 mg/kg. However, at 10 mg/kg 60% of AS2 rats survived the 30 wk experiment. All DA rats developed a high yield of adenocarcinoma of the bowel (means: males 6.8 tumours/large bowel and 0.8/small bowel; females 2.8/large and 0.08/small bowel) and one-third of the males developed tumours of the ear canal. A smaller proportion (52%) of HS rats developed tumours, specifically bowel tumours (means: male 1.4/large and 0.2/small bowel; females 0.4/large and 0.1/small bowel). Even on the lower dose of DMH AS2 rats showed extensive liver changes including cystic cholangioma (58%), angiosarcoma (25%) and hepatocellular carcinoma (8%) and 83% developed bowel tumours (means: males 2.3/large and 1.5/small bowel; females 3.5/large and 1.0/small bowel). The rapid induction and high yield of bowel tumours, the different toxicity of DMH in AS2 rats, and the differences in relative tumour density both between sexes and in the various segments of the bowel indicate the potential of these strains for studies of carcinogenesis in the bowel. PMID- 6272180 TI - Angiofibroma of the epididymis. PMID- 6272179 TI - Isoantigens A, B and H in primary liver malignancies. AB - Formalin-fixed tissues from 37 cases of primary liver malignancies were investigated by the technique of Specific Red Cell Adherence (SRCA) for studying the presence or absence of blood group isoantigens A, B and H. These tumours comprised 15 cases of hepatocellular carcinomas, 5 hepatoblastomas, and 17 cholangiocarcinomas. Results of the SRCA were consistently negative in all hepatocellular carcinomas and hepatoblastomas. Fifteen of 17 (88%) cholangiocarcinomas gave a positive SRCA, but no correlation could be obtained between the intensity of the reaction and their histological grading or metastatic potential. Reasons are given for the negative reactions in hepatocellular carcinomas and hepatoblastomas, an possible explanations are offered for the high rate of positivity in cholangiocarcinomas. It is concluded that the inherent properties of these 3 tumours limit the application of the SRCA procedure as a potential gauge of prognosis. PMID- 6272181 TI - [Changes in Na-K-ATPase activity in renal cortex and renal medulla experimental acute renal failure]. PMID- 6272182 TI - Urinary excretion of cyclic nucleotides and phosphate in response to parathyroid hormone and calcitonin in man. AB - The response to parathyroid hormone (PTH) and calcitonin (CT) was studied in eight children with various bone diseases by determining the serum calcium (Ca) and phosphate (P) concentration, urinary phosphate excretion rate, renal phosphate clearance, the percentage of filtered phosphate reabsorbed by the renal tubule (%TRP), creatinine clearance (Ccr), urinary cyclic adenosine 3',5' monophosphate excretion rate (UcGMPV). Administration of PTH caused no significant change in serum Ca and P values, whereas CT produced a decrease in Ca (delta Ca, -1.4 +/- 0.1 mg/100 ml) and P (delta P; -1.1 +/- 0.1 mg/100 ml). There was an increase in UcAMP V (delta UcAMP; 437 +/- 74 nmoles/min/100 ml Ccr) without any significant change in UcGMPV after administration of PTH. Phosphaturia was produced by both PTH (delta TRP, -18 +/- 3%) and CT (delta TRP, 13 +/- 2%). However, CT did not elicit any increase in either UcAMPV or UcGMPV. PMID- 6272183 TI - Developmental aspects of beta-adrenergic receptors and catecholamine-sensitive adenylate cyclase in rat myocardium. PMID- 6272184 TI - Abnormal growth kinetics and 5'-nucleotidase activities in cultured skin fibroblasts from patients with Duchenne muscular dystrophy. AB - The experiments reported herein compare growth kinetics and biochemical properties of cultured skin fibroblasts from patients with Duchenne muscular dystrophy (DMD) and matched normal controls. On day 7 after plating (6000 cells/cm2) cell number and DNA per dish are significantly reduced (P less than 0.0001) in the cultures from DMD patients (n = 14), compared to those from controls (n = 10). Moreover DMD cells contain less lipids and proteins per dish but more per cell than normal fibroblasts (not significant). Variations of media (McCoy's medium instead of Eagle's minimum essential medium) resulted in the same differences between DMD and control cells. Cell kinetic experiments (plating density: 2000 cells/cm2) show increased doubling times of DMD fibroblasts (P less than 0.001; nDMD = 5; ncontrols = 4) whereas plating efficiency is equal for both DMD and controls. On day 7 activity of the membrane bound enzyme 5'-nucleotidase either per mg protein or per microgram DNA is significantly elevated in cells from DMD patients (P less than 0.0005; nDMD = 8; ncontrols = 9) independent of cell density. Thus all findings in cultured DMD fibroblasts: increased doubling time, tendency to more voluminous cells, and elevated 5'-nucleotidase activity per cell suggest, that the DMD cells behave similar to prematurely aging cells. Until now we were not able to check whether any alterations of the plasma membrane are inducing early senescence or, reversely, premature aging is the cause of the postulated membrane alterations. If these findings were to be confirmed in cultured amniotic cells from DMD fetuses, thay could serve as a potential prenatal diagnosis of the disease. PMID- 6272185 TI - [X-ray computed tomography in the diagnosis of neoplasms in children]. PMID- 6272186 TI - [Advanced liver neoplasms in children]. PMID- 6272187 TI - [Problem of treatment of children with advanced neoplastic process]. PMID- 6272188 TI - [Primary liver cancer in the course of treatment of acquired aplastic anemia with oxymetholone in a 12-year-old girl]. PMID- 6272189 TI - [Pseudomembranous colitis. Possible relation with cytomegalovirus infection and influenza B]. PMID- 6272190 TI - The atp operon: nucleotide sequence of the promoter and the genes for the membrane proteins, and the delta subunit of Escherichia coli ATP-synthase. AB - The nucleotide sequence of the promoter region and the first five genes of the atp (or unc) operon of Escherichia coli has been determined. The first proposed gene in the operon contains four AUA codons and may be poorly expressed; it encodes a basic but yet hydrophobic protein which could function as a pilot protein for assembly of ATP-synthase. The three genes that follow are structural genes for proteins comprising the proton channel of the enzyme. The fifth gene codes for the delta-subunit of F(1)-ATPase. PMID- 6272191 TI - Novel topologically knotted DNA from bacteriophage P4 capsids: studies with DNA topoisomerases. AB - DNA molecules isolated from bacteriophage P4 are mostly linear with cohesive ends capable of forming circular and concatemeric structures. In contrast, almost all DNA molecules isolated form P4 tailless capsids (heads) are monomeric DNA circles with their cohesive ends hydrogen-bonded. Different form simple DNA circles, such P4 head DNA circles contain topological knots. Gel electrophoretic and electronmicroscopic analyses of P4 head DNA indicate that the topological knots are highly complex and heterogeneous. Resolution of such complex knots has been studied with various DNA topoisomerases. The conversion of highly knotted P4 DNA to its simple circular form is demonstrated by type II DNA topoisomerases which catalyze the topological passing of two crossing double-stranded DNA segments [Liu, L. F., Liu, C. C. & Alberts, B. M. (1980) Cell, 19, 697-707]. The knotted P4 head DNA can be used in a sensitive assay for the detection of a type II DNA topoisomerase even in the presence of excess type I DNA topoisomerases. PMID- 6272192 TI - Changes in chromatin structure at the replication fork. II The DNPs containing nascent DNA and a transient chromatin modification detected by DNAase I. AB - Discrete deoxyribonucleoproteins (DNPs) containing nascent and/or bulk DNA, were obtained by fractionating micrococcal nuclease digests of nuclei form 3H thymidine pulse (15-20 sec) and 14C-thymidine long (16 h) labeled sea urchin embryos in polyacrylamide gels. One of these DNPs was shown to contain the micrococcal nuclease resistant 300 bp "large nascent DNA" described in Cell 14, 259-267, 1978. The bulk and nascent mononucleosome fractions provided evidence for the preferential digestion by micrococcal nuclease of nascent over bulk linker regions to yield mononucleosome cores with nascent DNA. DNAase I was used to probe whether any nascent DNA is in nucleosomes. Nascent as well as bulk single-stranded DNA fragments occurred in multiples of 10.4 bases with higher than random frequencies of certain fragment sizes (for instance 83 bases) as expected from a nucleosome structure. However, a striking background of nascent DNA between nascent DNA peaks was observed. This was eliminated by a pulse-chase treatment or by digestion of pulse-labeled nuclei with micrococcal nuclease together with DNAase I. One of several possible interpretations of these results suggests that a transient change in nucleosome structure may have created additional sites for the nicking of nascent DNA by DNAase I; the micrococcal nuclease sensitivity of the interpeak radioactivity suggest its origin from the linker region. Endogenous nuclease of sea urchin embryos cleaves chromatin DNA in a manner similar to that of DNAase I. PMID- 6272193 TI - pUR222, a vector for cloning and rapid chemical sequencing of DNA. AB - A multipurpose plasmid, pUR222, was constructed. It contains six unique cloning sites (PstI, SalI, AccI, HindII, BamHI and EcoRI) in a small region of its lac Z gene part. Insertion of foreign DNA into the plasmid can be easily detected. Bacteria harbouring recombinant plasmids generally give rise to white colonies, while those containing only vector DNA form blue colonies on indicator plates. Plasmid DNA purified by a rapid method (Birnboim, H.C. and Doly, J. (1979) Nucl. Acids. Res. 7, 1513-1523) can be used for chemical sequencing of the cloned insert DNA. Labeled fragments need not be isolated after cutting with the proper restriction enzymes and are treated directly according to the sequencing protocol of Maxam and Gilbert. PMID- 6272194 TI - Characterization of a gene encoding a 115 K super T antigen expressed by a SV40 transformed rat cell line. AB - It has been reported that SV40-transformed V 11 F 1 clone 1 subclone 7 rat cells (subclone 7) produce a super T antigen of 115,000 M. This super T antigen is entirely SV40 coded and is synthesized by translation of an elongated form of SV40 early mRNA (May, E., Kress, M. Daya-Grosjean, L., Monier, R. and May, P. (1981) J. Virol., 37, 24-35). The results reported here show that there is only one independent insertion of viral DNA in the cellular genome of subclone 7 cells. When DNA from subclone 7 cells was cleaved with Bam HI endonuclease two distinct SV40 sequence containing fragments were generated with sizes of 5 Kb and 10 Kb, respectively. Two recombinant cosmids were constructed by insertion of the 5 Kb and 10 Kb fragments, respectively, into cosmid pHC 79. Using restriction map analysis and nucleotide sequencing, we showed that the 5 Kb fragment actually contained the complete sequence of a gene encoding super T antigen. As compared to the normal SV40 early gene, the sequence of super T gene showed the following rearrangements: (i) The segment between nucleotides 4116 - 3544 was duplicated in a direct order and (ii) these two copies of 573 nucleotide sequence were separated by a 93 nucleotide tract which was a nearly perfect inverted repeat of the segment located between nucleotides 4868 and 4776 (nucleotide numbering used here = Weissmann number +17). PMID- 6272195 TI - Nucleotide sequence of the lexA gene of Escherichia coli K-12. AB - A number of E. coli genes exhibit increased expression when the cellular DNA is damaged. In undamaged cells, lexA repressor limits the extent of their transcription, whereas, in damaged cells, the repressor is cleaved by a cellular protease, the product of the recA gene. We have sequenced 943 base pairs of cloned E. coli DNA containing the lexA gene. A regulatory region has been identified, followed by a translational open reading frame which encodes a polypeptide of 202 amino acids with a molecular weight of 22,300. The protein contains a single alanyl-glycyl peptide near its middle. This peptide is also found in certain phage repressors which are cleaved by the recA protease and has been shown to be the site of cleavage in these repressors. We have determined the nucleotide sequence of a portion of the lexA3 gene, whose product is 100-fold less susceptible to recA protease than the wild type repressor. We report a single base change (G to A) which alters the unique alanine-glycine sequence to alanine-aspartic acid. PMID- 6272196 TI - Organization and nucleotide sequence of a new ribosomal operon in Escherichia coli containing the genes for ribosomal protein S2 and elongation factor Ts. AB - We report the nucleotide sequence of the four min region of the Escherichia coli genetic map that includes the genes for ribosomal protein S2 (rpsB) and translation elongation factor EF-Ts (tsf), and the possible location of regulatory sites within this two gene operon. The data indicate that the gene order is: rpsBp-rpsB-tsf-tsft. One potential regulatory site is a 16 nucleotide sequence in the rpsB leader region encompassing the ribosome binding site and the translation initiation codon. This has a high degree of homology with nucleotides 8 through 23 on the 5' end of 165 ribosomal RNA, and might signify a sequence that is necessary for post-transcriptional control of rpsB expression. The data allow one to infer the amino acid sequences of S2 and EF-Ts. PMID- 6272197 TI - An improved procedure for utilizing terminal transferase to add homopolymers to the 3' termini of DNA. AB - Terminal deoxynucleotidyl transferase (E.C.2.7.7.3.1.) from calf thymus was used to add homopolymer tails to duplex DNA with 3' protruding, even, or 3' recessive ends. A gel electrophoresis method was employed to analyze the tail length and the percent of DNA with tails. In all the tailing reactions, dA, dT, and dC tails from CoCl2-containing buffer were longer than those from MnCl2 - or MgCl2 - containing buffers, whereas dG tails from MnCl2 -containing buffer were the longest. By varying the ratio of dNTP over DNA terminus and the concentration of terminal transferase, optimal conditions were found for adding dG or dC tails of 10-25 nucleotides in length and dA and dT tails of 20-40 nucleotides in length to duplex DNA with all types of 3' termini. PMID- 6272199 TI - Highly variable regions of DNA flank the human alpha globin genes. AB - A series of restriction fragment length polymorphisms which are due to DNA rearrangements have been identified within two highly variable regions flanking the human alpha globin genes. The existence of such highly polymorphic areas provides a large number of individual genetic markers for the alpha globin gene cluster on chromosome 16. If, as seems likely, such regions occur frequently throughout the human genome they should be of considerable value in the antenatal diagnosis of genetic disease. PMID- 6272198 TI - Reiterated sequences within the intron of an immediate-early gene of herpes simplex virus type 1. AB - We describe the nucleotide sequence of a herpes simplex virus type 1 DNA fragment containing the intron of the immediate-early mRNA-5 (IE mRNA-5) gene. The location of the intron within this fragment was determined by a Berk & Sharp nuclease S1 protection analysis, and by cloning and sequencing cDNA containing sequences overlapping t he IE mRNA-5 splice point. We found that the 149 base pair (bp) intron contained four copies of an identical 23 bp GC rich tandem repeat followed by a further reiteration consisting of the first 15 bp only. PMID- 6272200 TI - The conformation of adenovirus VAI-RNA in solution. AB - The secondary structure of an adenovirus associated low molecular weight RNA (VAI RNA) has been studied by partial digestion with T1-RNase and S1-endonuclease followed by T1-fingerprint analysis. The empirical secondary structure has been compared with two computer generated models based on minimal free energy of the structure. The results suggest that VAI-RNA in solution has a compact structure with a free energy of around -60 kcal with two stems and four bulge regions. The implication of this structure for the function of VAI-RNA is discussed. PMID- 6272201 TI - DNase II digestion of the nucleosome core: precise locations and relative exposures of sites. AB - The precise locations and relative exposures of the DNase II-accessible sites in the nucleosome core DNA are determined using techniques previously employed for the enzyme DNase I. It is found that there are a number of similarities between the site exposure patterns for the two enzymes but that in general the DNase II seems to discriminate less among adjacent sites' accessibilities than does DNase I. The two enzymes attack essentially the same positions in the DNA, the average difference between the precise location of the site being less than one-half base for the two enzymes. Such close similarities in the digestion patterns of two enzymes with such different mechanisms of scission show that the patterns reflect the structure of the nucleosome core and not merely the properties of the particular enzyme used. PMID- 6272202 TI - The helical periodicity of DNA on the nucleosome. AB - The precise number of base pairs per turn of the DNA double helix in the nucleosome core particle has been the subject of controversy. In this paper the positions of nuclease cutting sites are analysed in three dimensions. Using this midpoint of the DNA on the nucleosome dyad as origin, the cutting site locations measured along a strand of DNA are mapped onto models of the nucleosome core containing DNA of different helical periodicities. It is found that a helical periodicity of 10.5 base pairs per turn leads to cutting site positions which are sterically inaccessible. In contrast, a periodicity of 10.0 base pairs per turn leads to cutting site positions which are not only sterically sound, but which fall into a pattern such as would be expected when the access of the nuclease to the DNA is restricted by the presence of the histone core on one side and of the adjacent superhelical turn of DNA on the other. As proposed earlier by us (1), a value for the helical periodicity close to 10 base pairs per turn on the nucleosome, taken together with a periodicity close to 10.5 for DNA in solution - a value now established - resolves the so-called linkage number paradox. PMID- 6272203 TI - Transcription of the beta-like globin genes and pseudogenes of the goat in a cell free system. AB - We have examined the transcription in a cell-free system of all the members of a highly regulated gene family, the beta-like hemoglobin genes of the goat. These five genes, which code for embryonic, fetal, juvenile, and adult beta-globin proteins, are all transcribed to roughly the same extent in the in vitro system. In all cases initiation of transcription is accurate. However, two goat beta globin pseudogenes, as well as several artificially constructed deletion mutants, are not transcribed in vitro. A common feature of the transcriptionally inactive genes is the lack of an AT-rich consensus sequence just upstream of the presumptive initiation site. PMID- 6272204 TI - The nucleotide sequence of the replication origin of plasmid NTP1. AB - The sequence of the DNA of the origin region of NTP1 has been obtained. Analysis of the sequence indicates that: (1) there is great sequence homology in the DNA upstream from the origin in NTP1, ColE1, CLODF13, PBR345 AND PBR322; (2) only seven base pairs of NTP1 are identical with the sequence downstream from the origin in ColE1, although some homology exists for 140 bases downstream; (3) two ten base pair direct repeats are present in NTP1 which are also conserved in all four plasmids named above; (4) probably no polypeptide greater than fifteen amino acids in length is encoded by the NTP1 origin region, since no single open reading frame is conserved in all five plasmids. PMID- 6272205 TI - Structure and expression of a cloned beta o thalassaemic globin gene. AB - We have cloned the single beta-globin gene from an Italian patient who is a double heterozygote for beta o/delta beta o thalassaemia. RNA isolated from nucleated red cells from this patient can be translated in vitro to give detectable levels of A gamma- G gamma and alpha-globin, but no beta-globin. S1 mapping transcription studies show that beta-globin mRNA is present at about 1-3% of the level of alpha- and gamma-globin mRNA. In addition, the expression of this gene has been studied by reversed genetics. SV40-plasmid-beta o-globin gene recombinants have been transfected into Hela cells and analysed for beta-globin mRNA. In contrast to the results obtained with mRNA isolated directly from the blood of this patient, in the transfected Hela cells the same level of beta globin mRNA is seen for the beta o thalassaemic globin gene and for a normal beta globin gene. To elucidate the nature of the lesion, the entire DNA sequence of the beta-globin gene of this patient has been determined. The sequence shows that this gene contains a termination codon at position 39 (CAG - greater than UAG). Otherwise, there is a remarkable conservation of the entire DNA sequence. PMID- 6272206 TI - Sedimentation velocity of DNA in isokinetic sucrose gradients: calibration against molecular weight using fragments of defined length. AB - The relationship between sedimentation coefficient and molecular weight for DNA sedimenting in preformed alkaline and neutral sucrose gradients was determined using absolute molecular weight standards (restriction fragments of plasmid pBR322 and phage lambda DNA). The range of calibration for alkaline gradients was extended to small DNA fragments (652 base-pairs) for the first time. The exponent b in the equation S20 degrees, w = aMb was found to be 0.380 in neutral gradients and 0.410 in alkali. The latter value differs significantly from previous estimates. The gradients were isokinetic, and the distance sedimented was shown to be directly proportional to the sedimentation coefficient at all times. PMID- 6272207 TI - Identification of gene products programmed by restriction endonuclease DNA fragments using an E. coli in vitro system. AB - DNA restriction enzyme fragments have been used to programme the synthesis of polypeptides in an in vitro system without apparent loss in fidelity compared with supercoiled templates. The system is extremely sensitive, less than 1 microgram of DNA can be used to direct the synthesis of 35S-labelled polypeptides of sufficiently high specific activity such that products can be identified by SDS-PAGE after a few hours autoradiography. The ability to analyse fragments can be used to readily assign specific proteins to small regions of the coding template, to identify cloned gene products distinct from those of the vector, and to identify cloned genes expressed from their own promoters. The in vitro system can be used successfully with bacterial DNA from other species and efficient extracts can be prepared from any E. coli K-12 strain, which should greatly facilitate the purification of factors controlling the expression of specific genes by complementation assay. PMID- 6272208 TI - A general approach for purifying proteins encoded by cloned genes without using a functional assay: isolation of the uvrA gene product from radiolabeled maxicells. AB - The uvrA protein (UVRA) of E. coli has been extensively purified from a strain in which UVRA is overproduced and specifically labeled with 35S-methionine. This approximately 100-fold overproduction relative to normal strains is a result of having the uvrA gene present on a multicopy plasmid in a spr recA cell that makes defective lexA protein, the normal repressor of the uvrA gene, while the specific labeling of UVRA is done with maxicells. This approach facilitates the preparation of the protein since enzyme assays do not have to be carried out during the intermediate stages of purification. The purified UVRA binds to DNA and has ATPase activity but does not have intrinsic endonuclease activity. When added to extracts of uvrA- cells, the purified UVRA does promote the specific cutting of UV-irradiated DNA. Since this approach for working out rapid purification procedures by specifically labeling the proteins encoded by cloned genes does not require the use of a functional assay, it is a general one that can be applied to a wide variety of other gene products in addition to UVRA. PMID- 6272209 TI - Two new restriction endonucleases from Proteus vulgaris. AB - Two novel sequence-specific endonucleases have been isolated from Proteus vulgaris, ATCC 13315. PvuI recognizes the sequence: 5' C G A T decrease C G 3' 3' G C increase T A G C 5' and PvuII recognizes the sequence: 5' C A G decrease C T G 3' 3' G T C increase G A C 5' and cleave as indicated by the arrow (decrease). PvuI is an isoschizomer of XorII, RshI, and XniI. No enzyme with the specificity of PvuII has been described previously. PMID- 6272210 TI - Distribution of 5-methylcytosine in the DNA of somatic and germline cells from bovine tissues. AB - Genomic DNA of calf thymus contains 1.5 times as much 5-methylcytosine as similar sperm DNA, but the major EcoRI repeat fragment from satellite I of thymus contains ten times as much 5-methylcytosine as the corresponding fragment from sperm DNA. Restriction enzyme analyses of the total DNA and the satellite I fragment show that three HpaII sites in the fragment are completely unmethylated in sperm but fully methylated in thymus DNA. Under-methylation of many sites in the satellite DNAs can probably account for the lower level of methylation of sperm DNA rather than hemimethylation as previously suggested. These results are also discussed in relation to maintenance and de novo (initiation-type) methylases. PMID- 6272211 TI - Use of exonuclease III to determine the site of stable lesions in defined sequences of DNA: the cyclobutane pyrimidine dimer and cis and trans dichlorodiammine platinum II examples. AB - A method to detect chemically stable lesions in DNA has been developed using Exonuclease III, a double strand specific nuclease, to digest 5'-end labeled DNA. The products, when analyzed on high resolution DNA sequencing gels, reveal the sites of DNA modification. Cyclobutane pyrimidine dimers induced by UV irradiation can be localized by comparison of the fragments produced by Exonuclease III digestion with fragments obtained after digestion of the DNA with UV specific endonuclease. The experiments demonstrate the Exonuclease III stops one base away from the cyclobutane pyrimidine dimers. Similar experiments with cis- and trans-dichlorodiammine-platinum (II) showed that modification of DNA by these agents also impede Exonuclease III digestion. In general the same stop sites were found for cis-and trans-platinum adducts. They occur at sites of guanine bases. Additional stop sites were found for cis-platinum at sites of adjacent guanine bases. These results are in agreement with the model that cis platinum forms intrastrand guanine-guanine dimers, whereas trans-platinum does not. PMID- 6272213 TI - Variant surface glycoproteins of Trypanosoma brucei are synthesised with cleavable hydrophobic sequences at the carboxy and amino termini. AB - cDNAs coding for the amino and carboxy termini of two trypanosome variant surface glycoproteins (VSGs) have been sequenced. The results indicate that VSGs are synthesised with hydrophobic amino-terminal leader and carboxy-terminal tail sequences which are absent from purified mature VSGs. PMID- 6272212 TI - Vitellogenin in Drosophila melanogaster: sequence of the yolk protein I gene and its flanking regions. AB - We have isolated recombinant DNA clones coding for female specific proteins from Drosophila melanogaster. By screening with 32P-(A)+RNA from male and female flies, respectively, we were able to isolate a set of 100 cDNA clones which showed a positive hybridization signal for RNA from female flies. These clones have been rescreened with RNA isolated from fat body of two day old male and female flies. We obtained four positive cDNA clones. Isolation of the corresponding genomic sequences, construction of the physical map and comparing it with the restriction maps published by Barnett et al. (1) led us to conclude that we had isolated the genes coding for two of the three known yolk protein precursors (vitellogenins), YP I and YP II. The sequence of the YP I gene was determined. It gives rise to a protein of 48 700 dalton MW which might be cleaved to a MW of 46 700 during transport. The coding sequence is interrupted by a single intron of 75 bases in length. The proposed leader sequence starts at a region homologous to six heat shock gene sequences at the site of initiation of transcription, suggesting the existence of an 11 bp cap specific consensus sequence for Drosophila melanogaster. PMID- 6272214 TI - Characterization of nascent DNA fragments produced by excision of uracil residues in DNA. AB - Nascent short DNA chains could result from repair of incorporated uracil residues or be intermediates in discontinuous replication. We have characterized short DNA chains having apyrimidinic/apurinic-sites at 5' ends, the expected intermediates of repair, to distinguish them from RNA-linked replication intermediates. We have synthesized model substrates for the repair products; d(pRib[32P]poly(T)) and d(Rib[32P]poly(T)). Alkaline hydrolysis of both substrates has produced [5' 32P]poly(dT). Nascent short DNA was prepared from an Escherichia coli sof (dut) mutant, in this strain fragments from excision repair of uracil residues accumulate. The products of alkaline treatment are hardly digested by spleen exonuclease which selectively degrades 5'-hydroxyl-terminated DNA. These two results show that alkaline hydrolysis of the uracil repair fragments produces 5' phosphoryl-terminated DNA, whereas it is known that 5'-hydroxyl-terminated DNA is generated from RNA-linked DNA molecules. The two types of nascent fragments thus can be distinguished by the 5'-terminal structure produced by an alkaline hydrolysis. PMID- 6272216 TI - Interaction of histone H1 with superhelical DNA. Sedimentation and electron microscopical studies at low salt concentration. AB - Complexes of histones H1 with superhelical SV40 DNA obtained by direct mixing were studied in 0.1 SSC buffer corresponding to 0.02 M Na+. Depending on the molar input ratio H1/DNA three classes of sedimenting species were observed: (1) a component sedimenting similar to superhelical DNA with a sedimentation coefficient s2o,w of 25 S observable up to 335 Mol H1/Mol DNA (w/w = 2); (2) a component with s2o,w = 120 S appearing at 135 Mol H1/Mol DNA and (3) growing amounts of heterogeneous aggregates greater than 1000 S. Electron micrographs revealed the 25 S component to consist of double-fibers formed from one DNA molecule and the 120 S component to consist of bundles of several such double fibers. The aggregates represent cable-like structures. The addition of ethidium bromide to 25 S complexes induces the formation of bundles, if H1 is present in a quantity which alone is not sufficient to bring about this effect. This result indicates that ethidium bromide effects a redistribution of H1 molecules and that H1 is responsible for the bundle formation. PMID- 6272215 TI - Rifampicin-resistant initiation of DNA synthesis on the isolated strands of ColE plasmid DNA. AB - The opposite strands of the ColE1 and ColE3 plasmids were isolated as circular single-stranded DNA molecules. These molecules were compared with M13 and phi X174 viral DNA with respect to their capacity to function as templates for in vitro DNA synthesis by a replication enzyme fraction from Escherichia coli. It was found for both ColE plasmids that the conversion of H as well as L strands to duplex DNA molecules closely resembles phi X174 complementary strand synthesis and occurs by a rifampicin-resistant priming mechanism involving the dnaB, dnaC, and dnaG gene products. Restriction analysis of partially double-stranded intermediates indicates that preferred start sites for DNA synthesis are present on both strands of the ColE1 HaeII-C fragment. Inspection of the nucleotide sequence of this region reveals structural similarities with the origin of phi X174 complementary strand synthesis. We propose that the rifampicin-resistant initiation site (rri) in the ColE1 L strand is required for the priming of discontinuous lagging strand synthesis during vegetative replication and that the rri site in the H strand is involved in the initiation of L strand synthesis during conjugative transfer. PMID- 6272217 TI - The atp operon: nucleotide sequence of the genes for the gamma, beta, and epsilon subunits of Escherichia coli ATP synthase. AB - The nucleotide sequence of the promoter distal region of the atp (or unc) operon of Escherichia coli has been determined. It encodes the gamma, beta and epsilon subunits of the ATP-synthase complex and includes a noncoding sequence in which transcription of the operon probably terminates. This work completes the nucleotide sequence of the operon which contains nine genes: eight encode structural proteins of the ATP-synthase complex; a ninth, the first in the operon, may be a pilot for assembly. The genes for the alpha and beta subunits have evolved from a common ancestor. PMID- 6272218 TI - Sequence organization of the spacer in the ribosomal genes of Xenopus clivii and Xenopus borealis. AB - We have studied in X. clivii and X. borealis cloned EcoRI fragments containing the spacer located between the 28S and 18S ribosomal genes. We report for these two species the nucleotide sequences at both ends of the NTS region with special emphasis on the sequences around the transcription initiation site of the 40S rRNA precursor. In X. clivii the location of the 5' end of the precursor was mapped. In both species the sequences around the 40S origin are duplicated in the NTS. Nucleotide sequence comparison has revealed a stretch of 13 identical bases around the transcription initiation site of X. laevis and X. clivii. The same sequence is also present at the presumptive transcription initiation site of X. borealis rDNA. PMID- 6272219 TI - Location of single-stranded and double-stranded regions in rat liver ribosomal 5S RNA and 5.8S RNA. AB - Rat liver 5S rRNA and 5.8S rRNA were end-labelled with 32P at 5'-end or 3'-end of the polynucleotide chain and partially digested with single-strand specific S1 nuclease and double-strand specific endonuclease from the cobra Naja naja oxiana venom. The parallel use of these two structure-specific enzymes in combination with rapid sequencing technique allowed the exact localization of single-stranded and double-stranded regions in 5S RNA and 5.8 S RNA. The most accessible regions to S1 nuclease in 5S RNA are regions 33-42, 74-78, 102-103 and in 5.8 S RNA 16 20, 26-29, 34-36, 74-80 and a region around 125-130. The cobra venom endonuclease cleaves the following areas in 5S RNA: 7-8, 17-20, 28-30, 49-51, 56-57, 60-64, 69 70, 81-82, 95-97, 106-112. In 5.8S RNA the venom endonuclease cleavage sites are 4-7, 10-13, 21-22, 33-35, 43-45, 51-55, 72-74, 85-87, 98-99, 105-106, 114-115, 132-135. According to these results the tRNA binding sequences proposed by Nishikawa and Takemura [(1974) FEBS Lett. 40, 106-109], in 5S RNA are located in partly single-stranded region, but in 5.8S RNA in double-stranded region. PMID- 6272220 TI - Localization of DNA methyltransferase in the chromatin of Friend erythroleukemia cells. AB - Chromatin fragments released from intact Friend erythroleukemia cell nuclei during limited incubation with micrococcal nuclease, DNase II or DNase I were analyzed to determine the distribution of DNA methyltransferase in chromatin. The enzyme was released in a free form when internucleosomal DNA was digested with micrococcal nuclease but was found associated with Mg++-precipitable polynucleosomes after DNase II digestion. Less than 25% of the enzyme was released from nuclei incubated with DNase I under conditions where transcriptionally active chromatin should have been completely digested. These results indicated that the bulk of DNA methyltransferase was bound to "linker" DNA in condensed regions of chromatin. Preferential rebinding of free enzyme to linker DNA was also demonstrated in vitro. The possibility that chromatin proteins play a role in regulating access of DNA methyltransferase to specific sites in DNA is discussed in light of these findings. PMID- 6272221 TI - Protein synthesis and competitive ESR binding studies with E. coli ribosomes and spin-labeled polynucleotides. AB - Enzymatically prepared spin labeled copolymers of (U)n were tested for their ability to direct polyphenylalanine synthesis in vitro using E. coli B enzymes and ribosomes. Spin labeling of the C5 position using (RUGT,U)n (1:100) or (RUTT,U)n (1:100) did not alter the amount of polyphenylalanine formed in comparison to (U)n. In contrast, the C4 spin labeled copolymer (ls4U,U)n (1:100) reduced phenylalanine incorporation by 70-75% of the (U)n control levels. ESR monitoring of competitive ribosome binding to equimolar mixtures of polynucleotides was demonstrated with the macromolecular probe (DUTT,dT)n (1:100), the DNA analogue of (RUTT,U)n. The ESR competition approach showed that the affinity of the ribosomes was essentially the same for (dT)n, (A,U,G)n, and (A,U,G)n + tRNArmet. PMID- 6272222 TI - Enzymatic properties of the bacteriophage phi X174 A protein on superhelical phi X174 DNA: a model for the termination of the rolling circle DNA replication. AB - Incubation of phi X174 replication form I DNA with the A* protein of phi X174 in the presence of MN2+ results in the formation of three different types of DNA molecules: open circular form DNA (RFII), linear form DNA (RFIII) and the relaxed covalently closed form DNA (RFIV). The RFII and RFIII DNAs are shown to be A* protein-DNA complexes by electron microscopy using the protein labeling technique of Wu and Davidson (1). The linear double-stranded RFIII DNA molecule carries at one end a covalently attached A* protein whereas at the other end of the molecule the single-stranded termini are covalently linked to each other. The structure of the RFIII DNA shows its way of formation. The described properties of the A* protein indicate the way the larger A protein functions in the termination step of the rolling-circle type of phi X174 DNA replication. PMID- 6272223 TI - Requirement for the C-terminal region of middle T-antigen in cellular transformation by polyoma virus. AB - Deletions of polyoma virus DNA around the region that codes for the C-terminus of the viral middle T-antigen were created using a transforming fragment (BamH I/EcoR I) of viral DNA cloned in the plasmid vector pAT153. These species were recloned and assayed for their ability to transform Rat-1 cells in culture. Our results showed that whereas the DNA sequence between the presumed translational termination codon for the viral middle T-antigen and the single viral EcoR I site could be removed with no apparent effect on transformation, the removal of the termination codon itself or any amino acid coding sequences of this protein caused a drastic decrease in the transforming ability of the DNA. Transfection of Rat-1 cells with plasmids that contained viral DNA with deletions which corresponded to the last fourteen or more amino acids of the middle T-antigen never gave rise to cellular transformation. PMID- 6272224 TI - DNA sequence of the rat growth hormone gene: location of the 5' terminus of the growth hormone mRNA and identification of an internal transposon-like element. AB - The present communication describes the molecular cloning and DNA sequence determination of the rat growth hormone (rGH) gene. The rGH gene was cloned on an 11 kilobase EcoRI fragment of total rat DNA; it has four intervening sequences which correspond in position to those of the human growth hormone (hGH) gene. One of the intervening sequences in the rGH gene contains a possible transposable element: a 200 base pair direct repeat that is itself flanked by an exact 15 base pair direct repeat. The DNA sequence was used to estimate the location of the 5' end of the mature growth hormone mRNA. By S1 nuclease mapping it was located approximately 25 bases "downstream" from a TATAAA sequence presumed to play a role in initiation of transcription of the rGH gene. PMID- 6272225 TI - Anthramycin inhibition of restriction endonuclease cleavage and its use as a reversible blocking agent in DNA constructions. AB - Anthramycin can form a stable complex with DNA which does not dissociate upon repeated ethanol precipitations. The complex forms in less than one hour at pH 5.5. Bound anthramycin seems to be located in the minor groove of the DNA helix in the anthramycin DNA complex, since methylation of adenosine residues at N-3 by dimethylsulfate is reduced. The anthramycin-DNA complex is resistant to digestion by an excess of a number of restriction enzymes. Anthramycin can be removed from DNA by incubation at acid pH. The released DNA can then be cleaved by restriction enzymes. Anthramycin-DNA complexes can be acted upon by T4 polynucleotide ligase to form longer DNA molecules. The ability of anthramycin to form a stable but reversible complex which is not cleaved by restriction enzymes but can engage in joining reactions may allow a wider variety of DNA fragments to be more readily constructed in vitro. PMID- 6272226 TI - Nucleotide sequence of an Escherichia coli tRNA (Leu 1) operon and identification of the transcription promoter signal. AB - Fourteen different DNA fragments containing Escherichia coli tRNA genes have been cloned using the vector pBR322. We report the methods of cloning, the identification of specific tRNA genes, and the presence or absence of rRNA genes on these cloned DNA fragments. In particular, one chimeric plasmid contains a 17.0 kilobase pair EcoRI fragment bearing tRNA(Leu 1) sequences. Using nucleotide sequence analysis we have identified a cluster of three tandem tRNA(Leu 1) genes separated by intergenic spacers of 27 and 34 base pairs, respectively. The nucleotide sequence upstream of the first gene contains a transcription promoter site. A G-C rich sequence (5'-CGCCTCC-3') found between the Pribnow box and initiation site is very similar to the corresponding sequence found in other genes which are under stringent control. PMID- 6272227 TI - Structure of the baboon endogenous virus genome: cloning of circular virus DNA in bacteriophage lambda. AB - Linear, small and large circular forms of unintegrated viral DNAs were detected in Hirt supernatant fraction of human cultured cells infected with baboon endogenous virus M7. The circular M7 DNAs were cloned in bacteriophage lambda, Charon 28. Seventeen independent clones were isolated and analyzed by restriction endonuclease mapping. Nine clones were carrying a viral sequence of 8.6 kilobase pairs (kb) with two tandem repeats of 0.6 kb, which correspond to the large circular form of the unintegrated M7 DNA. Eight other clones had the viral insert of 8.0 kb, i. e., the small circular form, and were deleted one of the repeated sequences. The repeated sequences correspond to the long terminal repeats of 0.6 kb, located at both ends of the linear M7 DNA of 8.6 kb. One of the recombinants of the large circular M7 DNA had an inversion of 2.5 kb. One end of the inverted sequence was near the terminus of the long terminal repeats and the other in the gag gene region. The inversion seems to be occurred by integration of a viral DNA within itself during early periods of infection. The mechanism of the processes leading to integration is discussed from the structure of these unintegrated M7 DNAs as the precursors. PMID- 6272228 TI - The atp operon: nucleotide sequence of the region encoding the alpha-subunit of Escherichia coli ATP-synthase. AB - Part of the atp (or unc) operon encoding the alpha, beta, gamma, delta, and epsilon subunits of Escherichia coli ATP-synthase has been cloned into the plasmid pACYC 184. The DNA coding for the largest of these proteins, the alphas subunit, has been sequenced by cloning into the bacteriophage M13 and sequencing with dideoxy nucleotide chain terminators. It comprises 1539 nucleotides corresponding to a protein of 513 amino acids. PMID- 6272229 TI - Conformational stability of alternating d (CG) oligomers in high salt solution. AB - The conformation of d (CG)n oligomers with n = 2,3 has been studied in aqueous solution in the presence of high salt concentration. A minimum n value of three is necessary to obtain a left-handed Z-helix. When d (CG)3 is flanked by three non Z-helicogenic alternating AT sequences the left-handed helix is unstable and a B-type conformation is obtained also at high salt concentration. PMID- 6272230 TI - Pyrimidine arabinofuranosyl nucleosides with 5-substituted long, branched and unsaturated chains: synthesis and antiherpes properties. AB - Several procedures have been applied to the preparation of 5-alkyl analogues of araU and araC via condensation of the appropriate 2,4-bis-O-(trimethylsilyl)-5 alkylpyrimidine with 2,3,5-tri-O-benzyl-alpha-D-arabinofuranosyl chloride. The resulting O'-benzylated nucleosides were deblocked with the aid of BF3 . Et2O in C2H5SH. The araC analogues were also prepared by conversions of the corresponding 5-alkyl-araU derivatives. The chloromercuri derivatives of araU and araC, were reacted with allyl chloride in the presence of Li2PdCl4. The resulting 5-allyl derivatives were catalytically reduced to the corresponding 5-propyl analogues. Catalysed condensation of 2,4-bis-O-(trimethylsilyl)-5-vinyluracil with 2,3,5-tri O-benzyl-alpha-D-arabinosyl chloride, as well as with 1-O-acetyl-2,3,5-tri-O benzoylarabinofuranose, was carried out under a variety of different conditions. Deblocking of the benzylated nucleosides with various reagents led invariably to addition to the vinyl substituent. In the case of benzoylated nucleosides, deblocking yielded largely the alpha-anomers. The antiherpes activities of the 5 alkyl compounds have been evaluated, as well as the susceptibility of the araC analogues to enzymatic deamination. PMID- 6272231 TI - Further studies on the substrate specificity of calf spleen phosphodiesterase [EC 3.1.4.18]. AB - The synthesis of 5'-deoxy-5'-chlorothymidine-3'-(4-nitrophenyl)phosphate (5) and 5'-deoxy-5'-chlorothymidine-3'-(4-nitrophenyl)phosphorothioate (6) via corresponding phosphoranilidodiester intermediate is described. The affinity of 5 and 6 towards SPDE in comparison with thymidine-3'-(4-nitrophenyl)phosphate is tested. These findings reveal that the presence of 5'-hydroxyl function in the substrate is not necessary for hydrolytic action of this enzyme. PMID- 6272232 TI - Factors that affect patient compliance with psychiatric follow-up therapy after hospital discharge. PMID- 6272233 TI - Brachial plexus injury. PMID- 6272234 TI - [Serum ACTH and cortisol levels in bronchial asthma during treatment with Kenalog]. PMID- 6272235 TI - [Kinetics of plasma cyclic AMP level in atopic and non-atopic bronchial asthma after administration of triamcinolone acetonide]. PMID- 6272236 TI - Peptide hormones in liver cirrhosis and hepatocellular carcinoma. AB - Immunoreactive serum levels of human chorionic gonadotrophin (HCG), its alpha- and beta- subunits (alpha-HCG and beta-HCG), calcitonin (CT), parathyroid hormone (PTH), prolactin (PRL), adrenocorticotropic hormone (ACTH), and growth hormone (GH) were increased in 8 to 68% of 44 patients with hepatocellular carcinoma. With the exception of two patients, ACTH and PRL levels were only moderately increased, while alpha-HCG, GH, ACTH and PRL levels were not significantly different from the levels found in cirrhosis suggesting that metabolic effects due to impaired liver function may be responsible for their increase in liver cirrhosis and primary liver cell carcinoma. In contrast, HCG, beta-HCG, CT and PTH were associated with a higher incidence of elevated immunoreactive hormone levels than the other peptide hormones; higher concentrations were noted in tumor patients than with liver cirrhosis alone. Therefore, we suggest that metabolic effects due to cirrhosis may influence the serum levels and be more important than ectopic secretion by hepatocellular carcinoma. PMID- 6272237 TI - Concanavalin A binding of pregnancy-specific beta-1-glycoprotein in normal pregnancy and trophoblastic disease. AB - The binding of pregnancy-specific beta-1-glycoprotein (SP1) to Concanavalin A (Con A) has been studied in serum samples from 34 pregnant women and 13 patients with trophoblastic diseases. In normal pregnancy and in hydatidiform moles, the fraction that did not bind to Con A accounted for 0.5--4.3% of the total SP1 concentration whereas, in 5 out of 6 patients with choriocarcinoma, the fraction was 4.8--30%. The secretion of Con A non-binding SP1 appears to be an inherent characteristic of some malignant trophoblast tissues and may result from its altered glycosylation. PMID- 6272238 TI - [Practolol test for adrenergic beta receptor blockade in veterinary electrocardiographic diagnosis]. AB - The receptor theory of the action of catecholamines as well as the synthesis and production of beta-adrenolytics gave new perspectives for the treatment of circulatory diseases and enabled to use the beta-adrenergic blockade in electrocardiographic diagnosis. The aim of this work was to study whether it is possible to modify the oral method of administration used in human beings, into intravenous one, and what an information could be obtained as to actual heart condition after practolol injection. The results can be summarized as follows:- Practolol-Polfa injected intravenously in amounts of 30-50 mg for horse or cow and 3-10 mg for dog in ten minutes deviates the curve evoked by sympathectomy;- after the administration of the drug functional changes of final ventricular complex disappear which is the result of adrenergic system prevalence;- repolarization disturbances observed after the blockade indicate a myocardial defect and later, a partial decrease of changes in T wave shows that a vegetative factor is involved;--increased disturbances of the repolarisation phase after the practolol test indicate of myocardial defect due to compensatory mechanism of adrenergic system in relation to serious hemodynamic disturbances of the heart;- practolol test gives wider possibilities of interpretation, not only from the diagnostic point of view, but therapeutic as well. PMID- 6272239 TI - Are learning and attention related to the sequence of amino acids in ACTH/MSH peptides? AB - Learning and attention were examined in rats after injections of one of several molecules related to adrenocorticotropic hormone (ACTH) and melanocyte stimulating hormone (MSH). The initial phase of the learning process was linearly related to the length of the peptide with the smallest fragment (MSH/ACTH 4-10) improving learning the most and the largest molecule (ACTH 1-24) exerting no effect. Later stages of the learning problem, which were sensitive to the attentional state of the organism, resulted in U-shaped relations with the length of the same peptides. Enhancement of attention was significant only for the MSH compounds. These data indicate that some behaviors may be influenced as a function of the redundant information contained in the molecule while other behaviors may be discretely related to the specific conformation of the molecule. PMID- 6272240 TI - Naltrexone reduces weight gain, alters "beta-endorphin", and reduces insulin output from pancreatic islets of genetically obese mice. AB - Naltrexone, an opiate antagonist, was administered to young obese (ob/ob) and lean mice for five weeks. Animals had continuous access to food and received 10 mg/kg SC twice daily with equivalent volumes of saline given to controls. The effects on body weight, and pituitary and plasma levels of beta-endorphin-like material were measured. Naltrexone-injected obese animals gained weight more slowly over the first three weeks while the weight gain of lean animals was not affected by naltrexone. Plasma levels of beta-endorphin were shown to be significantly higher in untreated ob/ob mice and this difference increased with age (4-20 weeks). With naltrexone treatment, plasma levels in +/? mice rose and exceeded those in ob/ob. Saline treatment appeared to be a stress, and pituitary beta-endorphins rose 4-6 fold in ob/ob compared with +/?. While naltrexone reduced the levels of ob/ob pituitary towards normal, no effect on beta-endorphin levels in pituitary of lean mice was obtained. In vitro studies of effects of the opiate antagonists, naloxone, on insulin secretion by isolated islets provided additional evidence of resistance of lean mice to naloxone relative to ob/ob. (IRI secretion fell only in naloxone treated ob/ob islets). These observations support the contention that this form of genetic obesity is characterized by elevated endogenous opiate levels and an increased sensitivity to opiate antagonists such as naltrexone or naloxone. PMID- 6272241 TI - Endogenous opiates: 1979(1). PMID- 6272242 TI - Circadian placebo and ACTH effects on urinary cortisol in arthritics. AB - The effect of placebo and ACTH-1-17 (Synchrodyn, Hoechst) upon urinary free cortisol was examined at 5 different circadian stages on 10 men with Steinbrocker Stage II-III rheumatoid arthritis. A mean cosinor analysis of urinary cortisol data from the subjects prior to treatment with either ACTH or placebo revealed a statistically highly-significant rhythm. A circadian variation in a response of urinary free cortisol to a placebo was also seen. Moreover, the response of the midline-estimating statistic of rhythm (rhythm-adjusted circadian average) of urinary free cortisol to ACTH-1-17 by patients with rheumatoid arthritis is circadian rhythmic. This reactivity rhythm is out of phase with the spontaneous rhythm in urinary cortisol acrophases-in the tests limited thus far to midsummer. The further assessment of the circadian component in the context of broader interactions by rhythms with other frequencies in various conditions in health and disease is warranted by the demonstration of rhythms here presented for men with rheumatoid arthritis. PMID- 6272243 TI - Contrasting effects of Org 2766 and alpha-MSH on social and exploratory behavior in the rat. AB - Intraperitoneal injection of Org 2766 (0.01-0.4 microgram/kg) produced a dose related increase in the number of social contacts and in the time spent in active social interaction by pairs of male rats tested in arenas with which they were familiar, but had little effect when the rats were tested in unfamiliar arenas. The increased social interaction was not accompanied by any change in motor activity. In contrast, alpha-MSH (20-200 microgram/kg) decreased the time spent in active social dose-related. Both peptides reduced exploratory head-dipping only at high doses (4-8 microgram/kg for Org 2766 and 200 microgram/kg for alpha MSH); this change was not accompanied by a reduction in motor activity. PMID- 6272244 TI - Opiate binding properties of naturally occurring N- and C-terminus modified beta endorphins. AB - Beta-endorphin is further processed within the pituitary and brain by either N terminal acetylation, carboxy-terminal proteolysis, or both. These naturally occurring analogues are stored intracellularly and, in some tissues, represent the majority of beta-endorphin immunoreactivity detected by antisera. It is therefore critical to determine their relative potencies at the opiate receptor. This study demonstrates that cleavage of the C-terminus tetrapeptide brings about a 10-fold decrease in opiate binding potency of either camel or human beta endorphin. N-Acetylation, on the other hand, causes over a thousand fold loss in opiate potency rendering the peptide effectively inactive. Since unmodified beta endorphin is approximately equipotent at multiple opiate receptors, we tested for possible differential shifts towards mu or delta-type receptors which may result from the modification. Our results show no change in selectivity, but simply an overall loss of potency. PMID- 6272245 TI - Effects of beta-endorphin on body temperature in mice at different ambient temperatures. AB - The effect of intracerebroventricular injection of beta-endorphin (beta-END) on body temperature of mice was studied at ambient temperatures (Ta) of 10 degrees, 20 degrees and 31 degrees C. Doses between 0.1 and 10.0 microgram/mouse were studied. The lower (less than 1 microgram) doses of beta-END produced a hyperthermia at all Ta's studied. The higher doses of beta-END produced hyper- or hypothermia depending on the Ta. The subcutaneous injection of naloxone (1 mg/kg) antagonized the high dose hypothermic effects, but not the hyperthermic effect of beta-END. These data suggest that there may be different receptors and/or sites of action for high and low doses of beta-END. PMID- 6272246 TI - Minimum structure opioids-dipeptide and tripeptide analogs of the enkephalins. AB - Through a systematic reduction of peptide structure, a series of 25 tripeptide and 5 dipeptide amide and alcohol analogs of enkephalin were synthesized and assayed in vitro on the stimulated guinea pig ileum. Tyr-Pro-Phe-NH2, Tyr-D-Ala Phe-NH2, Tyr-D-Ala-Phe-ol and Tyr-D-Phe-Phe-NH2 had 20-25% the potency of Met enkephalin. Four aromatic alkylamides of the dipeptide Tyr-D-Ala were made with benzylamine, phenethylamine, phenylpropylamine and phenylbutylamine. All had full naloxone reversible enkephalin-like activity in the ileum assay. Tyr-D-Ala phenylpropylamide has about 80% the potency of Met-enkephalin in vitro, and is equipotent with Tyr-D-Ala-Gly-Phe-Met-NH2 in producing analgesia in mice after intraventricular administration. Tyr-D-Phe-NH2 is the smallest peptide to show full intrinsic enkephalin-like activity in vitro, although its potency is very low. PMID- 6272247 TI - beta-Endorphin-induced hyperthermia in the cat. AB - beta-Endorphin was injected into the third cerebral ventricle (ICV) of conscious, unrestrained cats. Hyperthermic response to 50 microgram of this peptide were reduced by 20-100 microgram naloxone given ICV 1 hr later. A dose of 40 microgram beta-endorphin increased body temperature at ambient temperature of 4, 22 and 34 degrees C, with the response being greater the warmer the environment. These results indicate that beta-endorphin acts on a central naloxone-sensitive receptor which is probably the v2 receptor that is activated by low doses of D Ala2-Met-enkephalinamide to evoke a similar pattern of change in body temperature over a comparable range of ambient temperatures. PMID- 6272248 TI - Laboratory diagnosis of viral hepatitis. PMID- 6272249 TI - Long-term use of potassium perchlorate. AB - A case of Graves' disease with potassium perchorate for 22 years without ill effect is described. Thyrotoxicosis recurred 4 weeks after the medication was withdrawn, suggesting that euthyroidism had been maintained by chronic use of the drug. As toxicity of perchlorate is probably dose related, it is suggested that long-term use of low dose perchlorate may be no more hazardous than alternative antithyroid therapy. PMID- 6272250 TI - Presynaptic receptors and modulation of the release of noradrenaline, dopamine and GABA. AB - A review of the role of presynaptic receptors in the modulation of neurotransmitter release indicates two types of presynaptic receptors. By means of presynaptic inhibitory autoreceptors a neurotransmitter can regulate its own release. In addition, presynaptic receptors are acted upon by other endogenous compounds, either transmitters released from adjacent nerve terminals, blood borne compounds or locally-formed substances, involved in trans-synaptic feed back. Both types of presynaptic receptors may be involved in the physiological control of transmitter release and are the target of drug action, either as agonists or as antagonists. PMID- 6272251 TI - Presynaptic receptors of central catecholamine neurones. AB - The article summarizes recent investigations on presynaptic alpha-adrenoceptors, opiate receptors and GABA receptors of central noradrenaline neurones, and on presynaptic dopamine receptors, opiate receptors and GABA receptors of central dopamine neurones. Pharmacological properties of the receptors, their location, and the steps linking receptor activation to changes in catecholamine release are discussed. In spite of a proliferation of research, there are large gaps in our knowledge of, in particular, GABA receptors and catecholamine release as well as opiate receptors and dopamine release. PMID- 6272252 TI - Beta-adrenergic receptors and responses in the heart. AB - This article summarizes some essential steps leading to one of the most prominent beta-adrenergic effects on the heart, the increase in force of contraction. Binding of beta-adrenergic agonists and antagonists, activation of the enzyme adenylate cyclase by agonist binding, increase in membrane permeability to calcium ions, possibly via cyclic AMP, and increase in force of contraction and its relation to enhanced calcium influx are briefly discussed. PMID- 6272253 TI - Receptor dynamics: effect of repeated administration of desmethylimipramine on the noradrenergic systems in rat brain. AB - The effects of a single and of repeated administration of desmethylimipramine (DMI) on the densities of various receptors and on the metabolism of noradrenaline (NA) were determined in rat brain. Inhibition of the uptake of NA into cortex slices, measured in vitro, was not diminished after repeated administration of DMI. The turnover of NA was inhibited only after the single administration of the drug. After repeated administration the turnover of NA was again within the normal range, but now the density of beta-receptors was significantly reduced. Repeated administration of DMI had no significant effect on the densities of alpha-adrenergic, dopaminergic, cholinergic, histaminergic, opiate and serotoninergic receptors. These results indicate that the continued blockade of NA-reuptake caused a selective desensitization of beta-receptor regulated systems in the brain. PMID- 6272254 TI - Subclasses of beta-adrenoceptors--a quantitative estimation of beta 1- and beta 2 adrenoceptors in guinea pig and human lung. AB - Binding studies have been performed in guinea pig and human lung membranes with the radioligand ICYP and several 'cardioselective' beta-adrenoceptor antagonists. Analysis of data from displacement curves by means of a curve fitting procedure, based on a non-linear regression analysis, indicated that in both tissues beta 1- and beta 2-type adrenoceptors coexisted. The relative proportions of the two receptor subtypes in a single tissue remained constant, irrespective of the antagonist used. The percentage of distribution between beta 1- and beta 2 adrenoceptors was about 22:78 in the guinea pig lung and 30:70 in human lung. For the compounds investigated, the affinities of beta 1- and beta 2-adrenoceptors correlated highly significantly between both species, suggesting that the 'cardioselectivity ratio" of a compound is the same in the guinea pig and in man. Mathematical treatment of the displacement curves provided no evidence of a further class of beta-adrenoceptors in lungs of both species. PMID- 6272255 TI - Electrical exploration of acetylcholine receptors. AB - A short account is given of the author's work on the neuromuscular junction, starting with the intracellular recording of spontaneous miniature endplate potentials and its simulation with a local microinjection of acetylcholine. The nature of quantal transmitter delivery and the molecular components of miniature endplate potentials are described as well as the effects of physical and chemical agents on the kinetics and the opentime of the channels of the endplate. The study of a variety of drugs has led to important advances in the electrical exploration of drug receptors and promises to shed new light on their mechanism of action and on the behaviour of cell membranes. PMID- 6272256 TI - A study of beta-adrenoceptors in the guinea pig. AB - The interaction of isoprenaline with propranolol and pindolol at beta adrenoceptors in isolated guinea pig atria, trachea and fat cells has been investigated. The affinity constants obtained revealed that in all three preparations both drugs acted as simple competitive antagonists. Although pindolol showed a somewhat greater affinity for atrial receptors than for those in the trachea neither of the antagonists showed any marked selectivity for adrenoceptors in either of these two preparations. In contrast, the affinity of fat cell receptors for either antagonist were markedly and consistently lower than those of the atrial or tracheal adrenoceptors. The theoretical implications of these results are discussed. PMID- 6272257 TI - Na+ channels in pancreatic islet cells. AB - Experiments are reviewed which show that Na+ channels which are pharmacologically identical with those found in spiking neurones are present in the outer membrane of pancreatic beta-cells. In both nerve and beta-cells the neurotoxins veratridine, scorpion toxin, and tetrodotoxin bind to three distinctly separate sites on the sodium channel, the affinity of each of these sites for its respective toxin being similar in the two tissues. The binding sites for veratridine and scorpion toxin exhibit reciprocal, positive, heterotropic cooperativity. The role of these Na+ channels in the physiological regulation of insulin release is discussed. PMID- 6272258 TI - Influence of an indigestible material on energy excretion by roosters and on true metabolizable energy of corn. AB - The influence of an indigestible and noncombustible material, silica gel (SG) on the excretion of energy by roosters and on the true metabolizable energy (TME) of corn was determined. Energy excretion by fasted roosters increased linearly with each increment of SG force-fed. The equation, energy excreted (kcal/24 hr) = 9.87 + (.47 x g SG) described this effect of SG. The data also showed that 24 hr was insufficient time for SG to completely clear the digestive tract, irrespective of amount force-fed. The TME of corn was 4.05 kcal/g dry matter when the grain was force-fed alone to roosters, when corn was force-fed in mixtures containing increasing proportions of SG, the TME of corn decreased with each increment of SG in the mixture. It was concluded that the extra energy excreted as a consequence of the presence of SG in mixtures with corn caused an underestimation of corn's TME. The regression coefficient relating energy excretion to amount of SG forced fed in the previous trial was used to correct energy excretion by roosters fed corn-SG mixtures, and corrected TME's for corn were calculated. The average corrected TME of corn was 3.97 kcal/g dry matter. These data illustrated that an indigestible material passing through the gastrointestinal tract of the chicken changed the amount of fecal metabolic energy excreted and, consequently, influenced the TME value of companion feedstuffs when the conventional TME assay procedure was used. PMID- 6272259 TI - A technique for the isolation of chicken hepatocytes and their use in a study of gluconeogenesis. AB - A new technique was developed for the isolation of chicken liver parenchymal cells. Glucose produced from 10 mM lactate was proportional to the amount of cells present. In the time-course study, gluconeogenesis from lactate and fructose was linear up to 60 min. Fructose proved to be the best substrate. Fructose was converted to glucose at the highest rate; this was followed by lactate, pyruvate, and xylitol. Alanine, glycerol, propionate, alpha ketoglutarate, and succinate proved to be poor substrates. There was no statistical difference between the results obtained with hepatocytes obtained from fed or fasted chickens. The isolated hepatocytes responded to glucagon, dibutyryl-cAMP, and epinephrine. The dose-response for glucagon was a sigmoid curve and the half-maximum stimulation was given by approximately 1 x 10(-2) micrometers hormone. The same type of curve was obtained with dibutyryl-cAMP, but the half-maximum stimulation was achieved at around 1.0 micrometer. The response to epinephrine was marginal. In the time-course experiment, prior to glucagon stimulation, glucose accumulated at a linear rate (slope = .2484). After the addition of the hormone, the level of cAMP increased by about 30% in the first minute and reached a peak (100%) in about 2 min; thereafter, it decreased to the level prior to the stimulation by the hormone. Two minutes after the addition of glucagon there was a significant increase in the rate of gluconeogenesis; this continued for another 3 min and then at a slower pace (slope = .2566). PMID- 6272260 TI - Effect of fiber on cholesterol metabolism in the Coturnix quail. AB - This experiment involving 288 Coturnix quail was conducted to determine the effects of various fiber sources (alfalfa, wheat bran, dried brewer's grain, cellulose, and pectin) on serum, liver and egg yolk cholesterol, and egg production. The fiber sources were added at a rate to provide 6.25% fiber to a corn-soybean meal diet and fed for a 28-day experimental period. Serum, liver, and egg cholesterol levels were measured as pen composite samples at the end of the experimental period. No difference was exhibited in egg yolk cholesterol among any of the groups (P greater than .10). Liver and serum cholesterol levels were elevated in the birds fed the pectin and wheat brain diets as compared with that in birds fed the other fiber source; also, metabolizable energy intakes and eggs per hen day (EHD) were decreased for both groups. When data were adjusted for EHD by covariance analysis, the treatment differences no longer appeared (P greater than .05). These results indicate that 1) there is a basal quantity of cholesterol deposited in the egg on which fiber intake, energy consumed, or egg production have very little effect and that 2) there is an inverse relationship between serum and tissue cholesterol levels and the total quantity of cholesterol excreted via the egg. PMID- 6272261 TI - Effect of dietary fiber sources on tissue mineral levels in chicks. AB - Dietary fiber may influence mineral status of chicks. Wheat bran, corn bran, soy bran, oat hulls, rice bran, and cellulose, were formulated into diets and fed to chicks for 4 weeks. Fiber-added diets contained additional 6% dietary fiber from the fiber sources. Body weights and feed intakes were recorded. Livers and tibias were analyzed for copper, zinc, iron, manganese, and magnesium. The rice bran diet resulted in significantly lowered body weights, feed intakes, and tibia concentrations of zinc, iron, and manganese. The other fiber-added diets did not produce significantly different values for the parameters examined. Dietary analyses showed that the rice bran diet did not have extremely high fiber levels as compared to the other diets. Addition of rice bran contributed the highest amount of phytic acid of all the fiber sources at 1.3% of the diet. It was concluded that when included at 6% of the diet, most fibers did not influence growth or tissue mineral levels. However, the addition of rice bran to the diet reduced body growth and deposition of zinc, iron, and manganese in tibias. It is possible that one fiber source may be beneficial while another is detrimental to nutritional status. PMID- 6272262 TI - Viability of and corticosteroid binding in lymphoid cells of various tissues after adrenocorticotropin injection. PMID- 6272263 TI - [Allergic reactions to food additives (author's transl)]. PMID- 6272264 TI - [Nutrition and cancer - an overview (author's transl)]. PMID- 6272265 TI - A new view of the etiology of nasopharyngeal carcinoma. PMID- 6272266 TI - [Comparative antirachitic activity of vitamins D2 and D3 in the body of chicks]. AB - In the four-week study the antirachitic action of vitamin D2 in different does (5000, 7500, 10 000 I. U. per kg food) and of vitamin D3 in a dose of 500 I. U. per kg food was investigated in chicks. The vitamin efficiency was measured with respect to the following seven parameters: weight gain, serum content of Ca and P, serum activity of alkaline phosphatase, bone ash content, content of Ca binding protein in the duodenal mucosa, and weight of parathyroid glands. In regard to these parameters, vitamin D2 in a dose of 7500 I. U./kg and vitamin D3 in a dose of 500 I.U./kg produced a similar effect. Thus, the antirachitic effect of vitamin D3 was 15 times higher than that of vitamin D2. In the three-day study of the effect of vitamins D2 and D3 on the synthesis of Ca-binding protein it was found that they exerted a similar effect when the dose of vitamin D2 was 14.5 times higher than that of vitamin D3. PMID- 6272267 TI - [Seroepidemiologic studies in lymphogranulomatosis]. PMID- 6272268 TI - [Liver function study in idiopathic myelofibrosis (subleukemic myelosis)]. PMID- 6272269 TI - Isolation and characterization of human actin genes. AB - We have utilized cloned actin genes from Drosophila melanogaster and from chicken to isolate 12 actin gene fragments from a human DNA library. Each of these 12 clones was shown to contain actin coding regions by its ability to selectively hybridize to human actin mRNA as assayed by in vitro translation. The translation product was judged to be actin on the basis of its comigration with authentic actins when electrophoresed on one- and two-dimensional NaDodSO4/polyacrylamide gels and on the basis of its partial proteolysis products. Determination of the sizes and order of the fragments generated by restriction endonuclease digestion of each of these recombinant phages allows us to conclude that they are nonallelic and are from nonoverlapping regions of the genome. We have used these cloned human actin genes and the Drosophila and chicken actin gene clones to show that the human genome contains 25-30 EcoRI fragments homologous to actin genes and that, among three nonconsanguineous individuals tested, none of these fragments exhibit length or restriction-site polymorphism. PMID- 6272270 TI - Molecular cloning and amplification of the adenylate cyclase gene. AB - A segment of DNA containing cya, the gene for adenylate cyclase [ATP pyrophosphate-lyase (cyclizing), EC 4.6.1.1], has been isolated from Salmonella typhimurium. The phage lambda gt4 was used as a cloning vector and adenylate cyclase-positive hybrid phages were isolated that complemented adenylate cyclase negative bacteria. The cloned DNA fragment encodes a polypeptide of molecular weight 81,000 that gives rise to adenylate cyclase activity. This protein represents a functional mutant of the bacterial adenylate cyclase. When the cya gene was amplified by inserting into a multicopy plasmid, the enzyme activity was overproduced 20-fold, but the cyclic AMP level increased only 60%, suggesting several probable regulatory mechanisms. Overproduction of enzymes by recombinant DNA techniques can be a useful probe of relationships in the metabolizing organism in vivo. PMID- 6272271 TI - Biosynthesis of phosphorylated forms of corticotropin-related peptides. AB - Phosphorylated forms of corticotropin[ACTH (1-39)], corticotropin-like intermediary lobe peptide[CLIP, ACTH (18-39)], and the common precursor for ACTH and beta-lipotropin (beta-LPH) have been identified in extracts of rat pituitaries, 32P-Labeled inorganic phosphate was successfully incorporated into ACTH (1-39), CLIP, and the ACTH/beta-LPH precursor in rat neurointermediary lobe explants and into ACTH (1-39) in isolated rat anterior pituitary cells. After peptidase digestion of the labeled CLIP and ACTH, the radioactive phosphate was recoverable as O-phosphoserine. The serine residue at position 31 was the only amino acid found to be phosphorylated in CLIP and ACTH (1-39). The unphosphorylated forms of both peptides were also synthesized. The demonstration of he incorporation of [32P]phosphate into CLIP, ACTH (1-39), and the ACTH/beta LPH precursor is consistent with the hypothesis that, within the rat intermediary lobe, phosphorylated CLIP is derived from a phosphorylated form of the common precursor, with phosphorylated ACTH (1-39) acting as a biosynthetic intermediate. PMID- 6272272 TI - Polarity of heteroduplex formation promoted by Escherichia coli recA protein. AB - When recA protein pairs circular single strands with linear duplex DNA, the circular strand displaces its homolog from only one end of the duplex molecule and rapidly creates heteroduplex joints that are thousands of base pairs long [DasGupta, C., Shibata, T., Cunningham, R. P. & Radding, C. M. (1980) Cell 22, 437-446]. To examine this apparently polar reaction, we prepared chimeric duplex fragments of DNA that had M13 nucleotide sequences at one end and G4 sequences at the other. Circular single strands homologous to M13 DNA paired with a chimeric fragment when M13 sequences were located at the 3' end of the complementary strand but did not pair when the M13 sequences were located at the 5' end. Likewise circular single-stranded G4 DNA paired with chimeric fragments only when G4 sequences were located at the 3' end of the complementary strand. To confirm these observations, we prepared fd DNA labeled only at the 5' or 3' end of the plus strand, and we examined the susceptibility of these labeled ends to digestion by exonucleases when joint molecules were formed. Eighty percent of the 5' label in joint molecules became sensitive to exonuclease VII. Displacement of that 5' end by recA protein was concerted because it did not occur in the absence of single-stranded DNA or in the presence of heterologous single strands. By contrast, only a small fraction of the 3' label became sensitive to exonuclease VII or exonuclease I. These observations show that recA protein forms heteroduplex joints in a concerted and polarized way. PMID- 6272273 TI - Theoretical aspects of translocation on DNA: adenosine triphosphatases and treadmilling binding proteins. AB - The basic kinetic and bioenergetic theory is outlined for two kinds of translocation on DNA: (i) helicases that use ATP to move along single-stranded DNA or to move on and invade double-stranded DNA at a replication fork; and (ii) DNA-binding proteins (not ATPases) that form bound aggregates on single-stranded DNA and facilitate replication by steady-state treadmilling of molecules between the ends of the aggregate. The respective resemblances to myosin--actin in muscle and to steady-state treadmilling in solution of actin or tubulin are pointed out. PMID- 6272274 TI - Histone-dependent and histone-independent forms of an ADP-ribosyltransferase from human and turkey erythrocytes. AB - An ADP-ribosyltransferase from turkey erythrocytes, which catalyzes the mono(ADP ribosylation) of guanidino compounds such as arginine and of many purified and crude cellular proteins, appears to exist both in high-activity, histone independent and low-activity, histone-dependent forms. At low salt concentrations, the activity of the transferase with agmatine as acceptor was less than 10% that observed in the presence of 200 mM NaCl. In the absence of salts, ADP-ribosylation of agmatine was stimulated greater than 10-fold by histones, and activity approached that observed with high salt concentration; under these conditions, the histones did not serve as ADP-ribose acceptors themselves. Histone also activated the highly purified ADP-ribosyltransferase from human erythrocytes. Enzyme activity was increased in the presence of salt and was then relatively independent of histones. DNA was not required for the stimulation of ADP-ribosylation by histone; incubation of the transferase and histone with DNase did not significantly decrease enzymatic activity. Additional DNA in the assay decreased the effect of histone. The erythrocyte ADP ribosyltransferase from diverse species thus appears to exist in two forms: one is dependent on histones for activity and one which, in the presence of salt, has high intrinsic activity and is independent of histone. The fact that the active forms of the transferase generated in the presence of salt or histone have similar catalytic activity suggests that these forms of transferase may be identical. It would appear that the enzymatic activity of transferase from different species may be controlled by histones. PMID- 6272275 TI - Interconverting mu and delta forms of the opiate receptor in rat striatal patches. AB - The binding of a radiolabeled "mu receptor" prototype opiate, dihydromorphine (H2morphine), and the binding of a "delta receptor" prototype, [D-Ala2,D Leu5]enkephalin (D-Enk), to slide-mounted rat caudate slices were simultaneously compared quantitatively and visualized by autoradiography. Generally, D-Enk bound to opiate receptors distributed evenly throughout the entire striatum (type 2 pattern), whereas H2morphine labeled discrete islands or patches of receptors (type 1 pattern). In the presence of Mn2+ (3 mM) or other divalent cations, however, Na+ and GTP at 25 degrees C caused an increase in D-Enk binding at the expense of H2morphine binding at striatal opiate receptor patches. Thus, these conditions shifted D-Enk binding from an even pattern to one that included both an even and patchy distribution. These incubation conditions not only promoted D Enk binding to striatal patches but also enabled the opiate receptor to regulate adenylate cyclase with the same (P less than 0.01) ligand selectivity pattern as that obtained by the displacement of D-Enk binding. The relative affinity of opiate receptors in striatal patches for opiate peptides, naloxone, and morphine appears to be a function of incubation conditions and coupling to adenylate cyclase and is not indicative of distinctly different opiate receptors. We postulate a three-state allosteric model consisting of mu agonist-, mu antagonists-, and adenylate cyclase-coupled delta-agonist-preferring states, whose equilibrium may be regulated by a sulfhydryl group mechanism. PMID- 6272276 TI - A proton gradient controls a calcium-release channel in sarcoplasmic reticulum. AB - Sarcoplasmic reticulum vesicles from mammalian skeletal muscle have previously been shown to develop a proton gradient (alkaline inside) of 0.15-0.5 pH units during active Ca2+ uptake. We found that dissipation of this gradient by the proton ionophores gramicidin, nigericin, and carbonyl cyanide p trichloromethoxyphenylhydrazone caused a rapid transient tension in skinned rabbit psoas muscle fibers. Increases, but not decreases, in medium pH of approximately 0.2 units over the range from pH 6.5 to pH 7.5 also elicited transient tensions. In isolated vesicles, physiological levels of Ca2+ (3.3 microM), inhibited pH-induced Ca2+ release. Dicyclohexylcarbodiimide blocked pH- and ionophore-induced Ca2+ release under conditions in which it could bind to sarcoplasmic reticulum proteins but did not inhibit Ca2+ uptake. We propose that a proton gradient generated across sarcoplasmic reticulum membranes during Ca2+ uptake maintains a Ca2+ release channel in a closed conformation and that dissipation of this gradient permits the Ca2+ release channel to open. We further propose that elevated myoplasmic Ca2+ also causes the Ca2+ channel to close, permitting Ca2+ uptake through Ca2+/Mg2+-ATPase to function effectively. As the proteolipids of sarcoplasmic reticulum bind dicyclohexylcarbodiimide under conditions in which Ca2+ release is blocked and as they have previously been shown to have Ca2+ ionophoric activity, we propose that the Ca2+-release channel either resides in the proteolipids or is controlled by H+ fluxes through the proteolipids. PMID- 6272277 TI - DNA flexibility studied by covalent closure of short fragments into circles. AB - The ring closure probability, or j factor, has been measured for DNA restriction fragments of defined sequence bearing EcoRI cohesive ends and ranging in size from 126 to 4361 base pairs (bp). The j factor is defined as the ratio of the equilibrium constants for cyclization and for bimolecular association via the cohesive ends. The end-joining reactions are fast compared to covalent closure of the cohesive ends by T4 DNA ligase. The rate of ligase closure is shown to be proportional to the equilibrium fraction of DNA molecules with joined cohesive ends, both in cyclization and in bimolecular association reactions. The j factor changes by less than 10-fold between 242 and 4361 bp, whereas it decreases by more than 100-fold between 242 and 126 bp as the DNA reaches the size range of the persistence length (150 bp). As regards ring closure, short DNA fragments are surprisingly flexible. These data are in good agreement with predictions by others for the ring closure probability of a wormlike chain. PMID- 6272278 TI - Reevaluation of the role of gangliosides in the binding and action of thyrotropin. AB - A cloned line of normal rat thyroid cells, FRTL, contained a small number of high affinity binding sites for thyrotropin (TSH) when measured under physiological conditions. The cells also bound small amounts of cholera toxin, and both hormone and toxin stimulated cyclic AMP production by the cells. The major ganglioside of FRTL cells was N-acetylneuraminylgalactosylglucosylceramide (GM3), with minor amounts of gangliosides corresponding to galactosyl-N-acetylgalactosaminyl-[N acetyl-neuraminyl]-galactosylglucosylceramide (GM1) and N acetylneuraminylgalactosyl-N-acetylgalactosaminyl-[N-acetylneuraminyl] galactosylglucosylceramide (GD1a). Treatment of these cells with neuraminidase (acylneuraminyl hydrolase, EC 3.2.1.18) converted most of the GD1a to GM1. After neuraminidase treatment, the binding of cholera toxin, which binds to GM1, was increased, but there was no change in the binding of TSH. Preincubation of neuraminidase-treated FRTL cells with the B (binding) component of cholera toxin completely prevented cholera toxin binding but had no effect on the binding of TSH. Neuraminidase treatment also somewhat enhanced, rather than decreased, the cyclic AMP response to TSH. Pretreatment of FRTL cells with mixed brain gangliosides resulted in a 10-fold increase in cholera toxin binding. Again there was no enhancement of TSH binding or adenylate cyclase stimulation. Finally, prolonged exposure of FRTL cells to TSH induced down-regulation of TSH receptors but had no effect on gangliosides or cholera toxin receptors. The results indicate that more complex gangliosides do not serve as a component of the TSH receptor nor are they involved in the transmission of the hormone signal across the cell membrane of these cultured rat thyroid cells. PMID- 6272279 TI - Rat alpha-lactalbumin has a 17-residue-long COOH-terminal hydrophobic extension as judged by sequence analysis of the cDNA clones. AB - cDNA for rat alpha-lactalbumin has been cloned in bacterial plasmid, and its sequence has been analyzed. The DNA sequence analysis shows that rat alpha lactalbumin has 17 extra residues beyond the COOH terminus of the alpha lactalbumin isolated and sequenced to date from other species. The predicted COOH terminal sequence is hydrophobic and proline rich and bears some resemblance to beta-casein sequences. PMID- 6272280 TI - On the molecular mechanisms of transposition. AB - We present a model for transposition that allows a choice between cointegrate formation (replicon fusion) and direct transposition. We propose that initiation of the process occurs by invasion of the target DNA by a single-stranded end of the transposable element. This leads to nicking of one of the DNA strands of the target molecule and ligation of this strand to that of the invading transposon. Transposition then occurs in a processive way by replication of the element from the invading end into the target site in a looped rolling-circle mode similar to replication of phage phi X174 replicative form to viral strand. The choice between cointegrate formation and direct transposition occurs at the nick ligation step, which terminates the process. We suggest that the choice is determined by the topology of the transposition enzymes and could be related to whether the element generates five- or nine-base-pair repeats in the target DNA on insertion. PMID- 6272281 TI - Antibodies specific for the polyoma virus middle-size tumor antigen. AB - We have obtained antibodies specific for the polyoma virus middle-size tumor antigen (middle T antigen) by immunizing rabbits with a synthetic peptide, Lys Arg-Ser-Arg-His-Phe, corresponding to the six carboxy-terminal amino acids of the middle T antigen predicted from the nucleotide sequence of polyoma DNA. The antipeptide serum precipitates the polyoma middle T antigen but not the small or large tumor antigens, and precipitation is inhibited in the presence of the peptide. Two cellular proteins, 30,000 and 26,000 daltons, are also precipitated specifically by the antipeptide serum and may have amino acid sequences related to the peptide. Two other cellular proteins, 33,000 and 25,000 daltons, are precipitated only in the presence of the peptide and may associate with it in cell extracts. Antisera directed against synthetic peptides are likely to be important in various ways, including the production of antibodies directed against particular determinants and the recognition of unknown proteins whose genes have been analyzed. PMID- 6272282 TI - Molecular cloning of poliovirus cDNA and determination of the complete nucleotide sequence of the viral genome. AB - The complete 7410 nucleotide sequence of poliovirus type I genome was obtained from cloned cDNA. Double-stranded poliovirus cDNA was synthesized and inserted into the Pst I site of plasmid pBR322, and three clones were derived that together provided DNA copies of the entire poliovirus genome. Two of the clones contained inserts of 2.5 and 6.5 kilobases and represented all but the 5' 115 bases of poliovirus RNA. A third clone was generated from primer-extended DNA and contained sequences from the 5' end of the viral RNA. An open reading frame that was identified in the nucleotide sequence starting 743 bases from the 5' end of the RNA and extending to a termination codon 71 bases from the 3' end contained known poliovirus polypeptide sequence. PMID- 6272283 TI - Identification and cloning of endogenous retroviral sequences present in human DNA. AB - Using nonstringent annealing conditions and a 2.75-kilobase segment of cloned African green monkey DNA that specifically hybridizes to the proviruses of AKR ecotropic murine leukemia virus (MuLV) and baboon endogenous virus (BaEV) as a probe, we detected related sequences in three different preparations of human brain DNA fragments. The blot-hybridization pattern obtained with cleaved human DNA was similar to that previously reported for the interaction of MuLV cDNA and cleaved mouse DNA and suggested the presence of numerous copies of retrovirus related sequences in the human genome. The labeled 2.75-kilobase fragment derived from cloned monkey DNA was used to screen a human DNA library in Charon 4A. One clone obtained hybridized to three contiguous MuLV-and BaEV-reactive fragments of the cloned monkey DNA and to multiple fragments of human DNA including a prominent 1.0-kilobase EcoRI fragment also present in the clone. PMID- 6272284 TI - Diphtheria toxin fragment forms large pores in phospholipid bilayer membranes. AB - The cytotoxic effect of diphtheria toxin requires the entry of its enzymatic A fragment (Mr approximately 21,000) into the cytosol of sensitive cells. We show that the B45 fragment (Mr approximately 24,000) forms, in lipid bilayers, pores that are large enough (diameter greater than or equal to 18 A) to allow the passage of extended fragment A. Pore formation is maximal when the B45-containing side is at low pH (4.7) and the opposite side is at high pH (7.4). These conditions resemble the pH gradient existing across lysosomal membranes. We suggest that fragment A passes through these pores from acidic endocytotic vesicles (lysosomes?) to the cytosol. PMID- 6272285 TI - Structural and motional changes in glyceraldehyde-3-phosphate dehydrogenase upon binding to the band-3 protein of the erythrocyte membrane examined with [15N,2H]maleimide spin label and electron paramagnetic resonance. AB - Binding of the glycolytic enzyme, glyceraldehyde-3-phosphate dehydrogenase [GAPDHase; D-glyceraldehyde-3-phosphate:NAD+ oxidoreductase (phosphorylating EC 1.2.1.12], to the cytoplasmic segment of band-3 protein in the erythrocyte (RBC) membrane has been examined by electron paramagnetic resonance (EPR) and saturation transfer EPR (ST-EPR) spectroscopies. GAPDHase, which was isolated from rabbit muscle and labeled with the resolution-enhancing deuterated N-(15N-1 oxyl-2,2,6,6-tetramethyl-4-piperidinyl)maleimide spin label ([15N,2H]MSL), showed the same binding specificity for the transmembrane band-3 protein of human erythrocyte membranes as reported for unlabeled GAPDHase from human RBC. Experimental EPR lineshapes from soluble and membrane-bound enzymes were analyzed by direct stimulation of spectra and indicated a structural alteration of the bound GAPDHase in the vicinity of the spin label, which was attached covalently to the active-site cysteine-149 residue. A rigorous theoretical analysis of the ST-EPR spectra of soluble and membrane-bound enzyme is presented and utilized in conjunction with model system analysis to demonstrate that the motion of membrane bound GAPDHase could be characterized by an effective isotropic rotational correlation time of 20 microseconds. This indicated that the GAPDHase--band-4 complex exhibits motional freedom relative to the membrane-spanning segment of the band-3 protein or the RBC. The double substituted spin label [15N,2H]MSL affords gains in sensitivity and resolution that permit studies of membrane-bound enzymes at physiological levels and quantitative simulations of the EPR and ST EPR lineshapes with reasonable computation times. PMID- 6272286 TI - Single K+ channel currents of anomalous rectification in cultured rat myotubes. AB - The currents through single K+ channels of the anomalous (or inward) rectifier were recorded in tissue cultured rat myotubes by using the "gigohm seal" patch clamp technique developed by Sigworth and Neher. These unitary currents were detected as current fluctuations due to the blocking and unblocking of channels by Ba2+. The single-channel conductance was obtained from the slope of the linear relationship between unitary current amplitude and membrane potential. When the external solution contained 155 mM K+, the single-channel conductance was 10.4 +/ 2.6 pS (+/- SD; n = 6). This value was independent of the the concentration of blocking ions but increased with increasing external K+ concentration. The behavior of the unitary current agreed with that expected from the blocking kinetics of Ba2+ on the macroscopic K+ current of the anomalous rectifier. The density of the channel is likely to be small and may even be less than 1/micrometers 2. PMID- 6272287 TI - The human beta-globin gene and a functional viral thymidine kinase gene in developing mice. AB - Two foreign cloned genes--one encoding a tissue-specific protein and one encoding a constitutive enzyme--were introduced into mouse eggs by microinjection into a pronucleus shortly after fertilization. They were the adult human genomic beta globin gene and the thymidine kinase (TK; ATP:thymidine 5'-phosphotransferase, EC 2.7.1.21) gene of herpes simplex virus (HSV), ligated in the pBR322 plasmid. Thirty-three developing mice were autopsied in late fetal life; all appeared normal. Blot hybridization tests revealed that the DNA of as many as five (15%) of the fetuses (from separate litters), and of their corresponding placentas, contained copies of the human beta-globin gene and of the HSV tk gene that had been retained and replicated without significant loss or rearrangement. The estimated total numbers of copies per cell were 3-50 for the donor globin gene and 3-20 for the donor tk. In some of the fetuses, these totals included some copies of molecular weight higher than that of the intact sequence; the additional segments may have arisen through changes such as deletions or duplications. The foreign genes in the five positive fetuses appear to be present in high molecular weight DNA. Assays capable of distinguishing between foreign and native TK indicated that at least one of the fetuses with the HSV tk gene had some TK enzyme of the HSV type and, therefore, at least one gene copy that was being accurately transcribed and translated to produce a functional protein, despite the absence of selective pressure. Thus, pure recombinant genes introduced into mice at the onset of their development can remain intact and be stably incorporated and even expressed. These experiments provide a practical basis for novel investigations of the developmental control of normal gene expression in vivo of the causes and possible cures of genetic diseases. PMID- 6272288 TI - Deoxyribonuclease I sensitivity of plasmid genomes in teratocarcinoma-derived stem and differentiated cells. AB - The DNase I (EC 3.1.21.1) sensitivities of the simian virus 40 (SV40) genome, the pBR322 genome, and the herpes simplex virus type 1 thymidine kinase (HSV-1 tk) gene have been compared in teratocarcinoma-derived stem (12-1) and differentiated (12-1a) cell lines established by transfection of thymidine kinase (ATP:thymidine 5'-phosphotransferase, EC 2.7.1.21)-deficient F9 cells with DNA from a tripartite plasmid genome consisting of the pBR322 genome, the SV40 genome, and the HSV-1 tk gene. HSV-1 tk is present in both stem and differentiated cells; SV40 early proteins are present in differentiated cells but not in stem cells; the pBR322 genome is not expressed in either cell type. The SV40 and pBR322 genomes are more sensitive to DNase I digestion in stem cells than in differentiated cells, reflecting the DNase I-hypersensitivity of total stem-cell chromatin. The HSV-1 tk gene is the least sensitive to DNase I digestion in both cell types. PMID- 6272289 TI - Direct identification of sickle cell anemia by blot hybridization. AB - Several reports have been published on the use of polymorphisms found in the human hemoglobin genes as a means for prenatal diagnosis of sickle cell anemia. The disadvantages of this approach reside in its limited application and the need for family analysis. Here we report that, by use of restriction endonuclease Dde I and diazobenzyloxymethyl-paper transfer procedures, a direct analysis can be made. Individuals with normal hemoglobin (AA) show two bands (175 and 201 base pairs) complementary to a 5'-specific beta-globin gene probe. Sickle cell trait individuals (AS) exhibit an additional band (376 base pairs). Individuals with sickle cell anemia (SS) show the band at 376 base pairs with a concomitant loss of the 175-base pair band. We interpret these changes in banding pattern to be the result of the elimination of a restriction site for Dde I in the altered codon associated with the sickle cell allele. Because an analysis can be performed on as little as 20 micrograms of cellular DNA, the application to prenatal diagnosis of sickle cell anemia should be possible. PMID- 6272290 TI - Identification and characterization of recombinant plasmids carrying the complete qa gene cluster from Neurospora crassa including the qa-1+ regulatory gene. AB - The early reactions in the catabolism of quinic acid in Neurospora crassa are controlled by at least four genes which are clustered on linkage group VII. Three of the loci (qa-2, qa-4, and qa-3) encode enzymes that convert quinic acid to protocatechuic acid. The fourth gene (qa-1) encodes a positive regulatory protein which, in the presence of quinic acid, leads to the de novo synthesis of the other proteins in the qa cluster. This communication describes a series of recombinant plasmids that span 36.5 kilobases of linkage group VII and contain the coding sequences for qa-2, qa-4, qa-3, and the qa-1 regulatory protein. The plasmids were obtained by partial digestion of wild-type N. crassa DNA with EcoRI and ligation into the cosmid cloning vehicle pHC79. Two independently derived plasmids (pMSK331 and pMSK335), each containing 36.5-kilobase inserts, were shown by transformation back into N. crassa to contain the entire qa gene cluster. A preliminary physical organization of the gene cluster is presented. An improved procedure for the transformation of N. crassa with plasmid DNA is also described. PMID- 6272291 TI - Increased susceptibility to cytomegalovirus infection in beige mutant mice. AB - Mice homozygous for the beige gene (bg/bg) are a homologue of the Chediak-Higashi syndrome of man and are known to be selectively defective in natural killer (NK) cells. We have compared the susceptibility of bg/bg and bg/+ C57BL/6J mice to infection with murine cytomegalovirus (MCMV). Beige mice are more susceptible to lethal infection and develop 33- to 43-fold higher virus titers in the liver, spleen, and kidney than do bg/+ mice after a sublethal infection, although virus replication is the same in vitro in cultured fibroblasts or epithelial cells from these mice. Inoculation with a sublethal dose of virus stimulates a NK cell response, although this is lower in bg/bg mice despite higher titers of interferon type 1 than in bg/+. A dose of MCMV that is lethal only to bg/bg augments cytotoxicity within 12 hr in bg/+ mice, whereas cytotoxicity in bg/bg remains very low. In bone marrow chimeras, recipients of bg/bg marrow were more susceptible to MCMV and had lower NK cell responses after virus inoculation than did recipients of marrow from bg/+ donors. The greater susceptibility of beige mice to the virus suggests that NK cells may contribute to resistance early in McMV infection. PMID- 6272292 TI - Homozygous familial hypercholesterolemia mutant with a defect in internalization of low density lipoprotein. AB - During studies on binding of low density lipoprotein (LDL) to fibroblasts from patients with the homozygous form of familial hypercholesterolemia (FH), a unique line was derived from subject M.N. This line could bind as much LDL as normal cells, or even more. However, like fibroblasts from other patients with the homozygous form of FH, it failed to show regulation of cholesterol synthesis. Analyses of the LDL receptors showed that this line could not mediate internalization of receptor-bound LDL. Studies on the fibroblasts of the parents of this subject showed that the inability to internalize LDL was hereditary and that the subject was a pure homozygote for this defect. The plausibility of this finding was supported by the fact that her parents were first cousins. The possible existence of a homozygous state of this defect was predicted by Goldstein et al. [Goldstein, J. L., Brown, M. S. & Stone, N. J. (1977) Cell 12, 629-41], but an actual case of the internalization defect in a pure homozygous form had not been found. There were no differences from normal cells in the nature of the LDL binding activity of this line, such as in its specificity, affinity, or Ca2+ requirement. PMID- 6272293 TI - Activation of related transforming genes in mouse and human mammary carcinomas. AB - High molecular weight DNAs of five tumors induced by mouse mammary tumor virus (MMTV), two mouse mammary tumors induced by a chemical carcinogen, and one human mammary tumor cell line (MCF-7) were assayed for the presence of transmissible activated transforming genes by transfection of NIH 3T3 mouse cells. DNAs of all five MMTV-induced tumors, one chemical carcinogen-induced tumor, and the human tumor cell line induced transformation with high efficiencies (approximately 0.2 transformant per micrograms of DNA). NIH cells transformed by DNAs of MMTV induced tumors did not contain exogenous MMTV DNA sequences, indicating that MMTV induced mammary carcinomas contained activated cellular transforming genes that were not linked to viral DNA. The transforming activities of DNAs of all five MMTV-induced tumors, the chemical carcinogen-induced mouse tumor, and the human tumor cell line were inactivated by digestion with the restriction endonucleases Pvu II and Sac I, but not by BamHI, EcoRI, HindIII, Kpn I, or Xho I. These results indicate that the same or closely related transforming genes were activated in six different mouse mammary carcinomas, induced by either MMTV or a chemical carcinogen, and in a human mammary carcinoma cell line. PMID- 6272294 TI - Differential immunologic reactivity and processing of glycoproteins gA and gB of herpes simplex virus types 1 and 2 made in Vero and HEp-2 cells. AB - Herpes simplex virus types 1 and 2 (HSV-1 and HSV-2, respectively) specify five major glycoproteins designated as gA, gB, gC, gD, and gE. Previous studies have shown that gA and gB differ in electrophoretic mobility but not in reactivity with antisera prepared to each of these glycoproteins. Moreover, gA and gB of HSV 1 crossreact in serologic tests with the corresponding glycoproteins of HSV-2. In this paper, we report on the reactivities of gA and gB of HSV-1 and HSV-2 with 24 independently derived monoclonal antibodies reactive with these antigens. Our results show the following: (i) Electrophoretic mobilities of HSV-1 and HSV-2 glycoproteins gA and gB made in HEp-2 cells are significantly less than those made in Vero cells. (ii) All monoclonal antibodies precipitated both gA and gB made in infected HEp-2 cells. These include 12 monoclonal antibodies that neutralized virus and 12 that did not. (iii) HSV-2 glycoproteins gA and gB made in HEp-2 cells contain type-specific domains. However, monoclonal antibodies produced by one clone directed to these domains did not react with glycoproteins made in Vero cells. (iv) Lysates of infected Vero cells contain three lower molecular weight polypeptides that also reacted with monoclonal antibodies directed to glycoproteins gA and gB. These polypeptides are virus specific inasmuch as those specified by HSV-1 differ in electrophoretic mobilities from those specified by HSV-2. these polypeptides are absent in lysates of HEp-2 cells. PMID- 6272295 TI - Baboon endogenous virus genome: molecular cloning and structural characterization of nondefective viral genomes from DNA of a baboon cell strain. AB - Several heterogeneities in the baboon endogenous virus (BaEV) genomes that are present in the DNA of normal baboon tissues and the baboon cell strain BEF-3 have been described previously. To study these genomes, we cloned BaEV proviruses from BEF-3 cellular DNA into the lambda vector Charon 4A. Of the four full-length clones isolated, one was nondefective as determined by transfection. The sequence of a portion of this clone was found to code for amino acids 61-91 in the p30 region of the gag gene. This identification allowed us to align the restriction map with the BaEV genetic map. One heterogeneity, a BamHI site 2.4 kilobases (kb) from the proviral 5' end, was located close to the gag-pol junction; another, a BamHI site 1.4 kb from the 5' end of the genome, corresponded to the gag p30 coding sequence for amino acids 32-34; and a third, a Xho I site, was near the 3' end of the pol gene. To select the nondefective BaEV genomes from BEF-3 cells, we infected permissive cells with virus produced by BEF-3 cells and also transfected BEF-3 cellular DNA into permissive cells. The BaEV genomes in the permissive recipient cultures were then analyzed by restriction enzyme analysis. These nondefective genomes were found to be heterogeneous with respect to the gag-pol BamHI site and the Xho I site, but all were found to contain the BamHI site 1.4 kb from the 5' end of the genome. PMID- 6272296 TI - Morphine and opioid peptides reduce inhibitory synaptic potentials in hippocampal pyramidal cells in vitro without alteration of membrane potential. AB - We used intracellular recording in the hippocampal slice in vitro to characterize further the mechanisms behind the unusual excitatory action of opiates and opioid peptides on hippocampal pyramidal cells in vivo. No significant effect on resting membrane potential, input resistance, or action potential size in cortical area 1 (CA1) pyramidal cells was observed with morphine sulfate, beta-endorphin, [Met5]enkephalin, or [D-Ala2, D-Leu5]enkephalin at 1-50 microM. However, in all cells studied, these agents markedly reduced the size of inhibitory postsynaptic potentials generated by stimulation of the stratum radiatum or alveus. Excitatory postsynaptic potentials were also diminished in many of these cells. The effects of the opioids were antagonized by naloxone. These results are consistent with excitation of pyramidal neurons by a disinhibitory mechanism. PMID- 6272297 TI - Differentiation of the open and closed states of the ionic channels of nicotinic acetylcholine receptors by tricyclic antidepressants. AB - The actions of two clinically important dibenzocycloheptane antidepressant drugs, amitriptyline and nortriptyline, were studied on ionic channels of nicotinic acetylcholine (AcCho) receptors at the neuromuscular junction of frog skeletal muscle. Amitriptyline (5-10 microM) and nortriptyline (1-2 microM), like imipramine (5-10 microM), did not react with the nicotinic AcCho receptor but caused a voltage- and time-dependent decrease in the peak amplitude of the endplate current (epc). The time constant of epc decay, however, retained its voltage sensitivity. The voltage- and time-dependent effect of amitriptyline was nonlinear with regard to the current/voltage (I/V) relationship. Nortriptyline also had a more pronounced voltage- and time-dependent effect evidenced by a hysteresis loop in the I/V relationship of the epc was eliminated by the use of 50-msec stepwise changes of the membrane potential. The nonlinearity and hysteresis were due to a time-dependent phenomenon and did not involve previous AcCho receptor activation. The rate constant of the voltage- and time-dependent decrease in epc amplitude was sensitive to the membrane electric field and varied linearly with the membrane potential. Iontophoretically elicited epcs were much more depressed by both drugs than were spontaneous miniature epcs. There was no effect on the time constant of miniature epc decay, single-channel lifetime, or conductance. Thus (as we have pointed out in our histrionicotoxin studies) the primary site of action of these agents presumably is the activated but nonconducting species of the ionic channel of the nicotinic AcCho receptor. These agents, particularly nortriptyline, point to several different binding sites of the ionic channel and are suitable tools for the separation of the effects on peak current amplitude from its time constant of decay. PMID- 6272298 TI - Potassium current suppression by quinidine reveals additional calcium currents in neuroblastoma cells. AB - Quinine and quinidine have been evaluated with regard to their effects on the electrical activity of neuroblastoma cells. Under voltage-clamp conditions, we have found that quinine and quinidine block both the voltage-dependent and Ca2+ dependent K+ conductances. Blockage of the voltage-dependent K+ channel is manifest as an increase in the amplitude and in the duration of the action potential. Blockage of the Ca2+-dependent K+ channel in Na+-free (replaced by Tris) solutions containing 6.8 mM Ca2+ and tetraethylammonium ion or 4 aminopyridine (to block the voltage-dependent K+ current) is seen as a further prolongation of the Ca2+ action potential and diminution of the after hyperpolarization. A critical role of the Ca2+-dependent K+ conductance in modulation of the rate and duration of trains of Ca2+ action potentials is shown by the use of low concentrations (5-40 microM) of quinine or quinidine, which diminish the Ca2+-dependent K+ conductance in a graded manner. After complete blockade of K+ currents, the peak Ca2+ currents are enhanced at all voltages, especially at values more positive than -30 mV, where a steady-state inward current appears as well. In this same voltage range, the decay of the Ca2+ current exhibits two time constants--that of the transient inward current, which is about 20 msec, and a much slower (approximately 2000 msec) component. It is suggested that neuroblastoma cells have two types of calcium channels--one which generates the Ca2+ action potential and a second, distinguished by activation at more depolarized levels and by a slow rate of inactivation, which underlies the calcium entry necessary to activate the Ca2+-dependent K+ conductance. PMID- 6272299 TI - Nucleotide sequence of the mRNA encoding the pre-alpha-subunit of mouse thyrotropin. AB - We have constructed and cloned in bacteria recombinant DNA molecules containing DNA sequences coding for the precursor of the alpha subunit of thyrotropin (pre TSH-alpha). Double-stranded DNA complementary to total poly(A)+RNA derived from a mouse pituitary thyrotropic tumor was prepared enzymatically, inserted into the Pst I site of the plasmid pBR322 by using poly(dC).poly(dG) homopolymeric extensions, and cloned in Escherichia coli chi 1776. Cloned cDNAs encoding pre TSH-alpha were identified by their hybridization to pre-TSH-alpha mRNA as determined by cell-free translations of hybrid-selected and hybrid-arrested RNA. The nucleotide sequences of two cDNAs (510 and 480 base pairs) were determined with chemical methods and corresponded to much of the region coding for the alpha subunit and the 3' untranslated region of pre-TSH-alpha mRNA. The sequence of the 5' end of the mRNA was determined from cDNA synthesized by using total mRNA as template and a restriction enzyme DNA fragment as primer. Together these sequences represented greater than 90% of the coding and noncoding regions of full-length pre-TSH-alpha mRNA, which was determined to be 800 bases long. The amino acid sequence of the pre-TSH-alpha deduced from the nucleotide sequence showed a NH2-terminal leader sequence of 24 amino acids followed by the 96-amino acid sequence of the apoprotein of TSH-alpha. There is greater than 90% homology in the amino acid sequences among the murine, ruminant, and porcine alpha subunits and 75-80% homology among the murine, equine, and human alpha subunits. Several regions of the sequence remain absolutely conserved among all species, suggesting that these particular regions are essential for the biological function of the subunit. The successful cloning of the alpha subunit of TSH will permit further studies of the organization of the genes coding for the glycoprotein hormone subunits and the regulation of their expression. PMID- 6272301 TI - Cloning of bacteriophage fd gene 2 and construction of a plasmid dependent on fd gene 2 protein. AB - Bacteriophage fd gene 2 was cloned in plasmid pBR325. Cells carrying the hybrid plasmid produce about 200 times more enzymatically active fd gene 2 protein than did cells infected with phage fd wild type, as measured by replication of phage fd replicative form I in vitro. Cloned gene 2 supports replication of an artificial phage fd miniplasmid consisting of the origin of bacteriophage fd replication and a gene coding for kanamycin resistance. This plasmid occurs in high copy numbers and is viable only in cells carrying the cloned fd gene 2 or in cells infected with phage fd. Because the miniplasmid is not propagated in natural hosts, it can be considered a safe cloning vector. Its fusion with the gene 2 hybrid plasmid provides an autonomous replicon independent of the polA function of the host cell. fd gene 2 is the only phage-encoded trans-acting function required for replication of double-stranded fd DNA in vivo. PMID- 6272300 TI - Specific gangliosides function as host cell receptors for Sendai virus. AB - The ability of specific gangliosides to function as host cell receptors for Sendai virus was investigated by using Madin-Darby bovine kidney cells which become resistant to infection upon treatment with Vibrio cholerae sialidase. Sialidase-treated cells were incubated for 20 min at 37 degrees C with individual, highly purified gangliosides containing homogeneous carbohydrate moieties and then inoculated with virus for 10 min. Susceptibility of the cells to infection was monitored by hemagglutination titer of the virus produced 48 hr after inoculation. Incubation of the cells with gangliosides containing the sequence NeuAc alpha 2,3Gal beta 1,3GalNAc (i.e., GD1a, GT1b, and GQ1b) fully restored susceptibility to infection to the cells. However, the ganglioside GQ1b in which the sequence ends with two sialic acids in a NeuAc alpha 2,8NeuAc linkage instead of a single sialic acid as in GD1a and GT1b, was effective as a receptor at a concentration 1/100th that of any of the other gangliosides tested. Incubation with gangliosides similar in structure to GD1a, GT1b, and GQ1b but lacking the sialic acid attached to the terminal galactose (i.e., GM1 and GD1b) had no effect. The results from control experiments in which gangliosides were incubated at 0 degrees C with cells or in which trypsin was used to remove gangliosides adsorbed to cells were consistent with the premise that the gangliosides must actually insert into the cellular membrane to function as Sendai virus receptors. Addition of 4 X 10(6) molecules of 14C-labeled GD1a per cell made the cells fully susceptible to infection. Analysis of the ganglioside content of cell membranes showed that gangliosides GD1a, GT1b, and GQ1b are natural components of these cells and are present in quantities sufficient to act as receptors. These results demonstrate that gangliosides with the proper carbohydrate sequence, such as GD1a, GT1b, and GQ1b, function as natural receptors for Sendai virus in host cells. PMID- 6272302 TI - Gene II of phage f1: its functions and its products. AB - Plasmids harboring the amino-terminal part of bacteriophage f1 gene II confer to bacterial cells partial resistance to infection with the male-specific bacteriophages f1 and f2. This effect (IP-2 phenotype) is due to the production of large amounts of an approximately 20,000-dalton polypeptide corresponding to the amino-terminal part of gene II protein. These results have allowed the isolation of clones producing functional gene II protein in large amounts. An in vitro assay has been developed to test the enzymatic activity of the gene II protein produced. PMID- 6272303 TI - Both alpha and beta subunits of human choriogonadotropin photoaffinity label the hormone receptor. AB - It has been shown that a photoactivable derivative of human choriogonadotropin (hCG) labels the lutropin receptor on porcine granulosa cells [Ji, I. & Ji, T. H. (1980) Proc. Natl. Acad. Sci. USA 77, 7167-7170]. In an attempt to identify which of the hCG subunits labeled the receptor, three sets of different hCG derivatives were prepared. In the first set, hCG was coupled to the N-hydroxysuccinimide ester of 4-azidobenzoylglycine and radioiodinated. In the second set, only one of the subunits was radioiodinated, but both subunits were allowed to react with the reagent. In the third set, both the reagent and [125I]iodine were coupled to only one of the subunits. The binding activity of each hormone derivative was comparable to that of 125I-labeled hCG. After binding of these hormone derivatives to the granulosa cell surface, they were photolyzed. After solubilization, autoradiographs of sodium dodecyl sulfate/polyacrylamide gels of each sample revealed a number of labeled bands; the hCG derivatives containing 125I-labeled alpha subunit produced four bands (molecular weights 120,000 +/- 6,000, 96,000 +/- 5,000, 76,000 +/- 4,000, and 73,000 +/- 4,000) and those containing 125I-labeled beta subunit produced three bands (molecular weights 106,000 +/- 6,000, 88,000 +/- 5,000, and 83,000 +/- 4,000). Results were the same when the hormone-receptor complexes were solubilized in 0.5% Triton X-100 and then photolyzed or when the hormone was derivatized with a family of reagents having arms of various lengths. We conclude that both the alpha subunit and the beta subunit of hCG photoaffinity labeled certain membrane polypeptides and that these polypeptides are related to the hormone receptor. PMID- 6272304 TI - Staphylococcal alpha-toxin: oligomerization of hydrophilic monomers to form amphiphilic hexamers induced through contact with deoxycholate detergent micelles. AB - Native staphylococcus aureus alpha-toxin is secreted as a hydrophilic polypeptide chain of Mr 34,000. The presence of deoxycholate above the critical micellar concentration induced the toxin monomers to self-associate, forming ring or cylindrical oligomers. The oligomers were amphiphilic and bound detergent. In deoxycholate solution, the protein-detergent complexes exhibited a sedimentation coefficient of 10.4 S. A Mr of 238,700 was determined by ultracentrifugation analyses at sedimentation equilibrium. Because quantitative detergent-binding studies indicated a protein/detergent ratio of approximately 5:1 (wt/wt), the protein moiety in each protein-detergent complex was determined to be approximately Mr 200000, corresponding to a hexamer of the native molecule. The amphiphilic toxin hexamers were ultrastructurally indistinguishable from the cytolytic, annular toxin complexes that form on and in biological target membranes. They bound lipid and could be incorporated into artificial lecithin lipid vesicles. The transition of toxin protein molecules from a hydrophilic monomer to an amphiphilic oligomer through self-association has thus been shown to be inducible solely through contact of the native protein molecules with an appropriate amphiphilic substrate. PMID- 6272305 TI - Functional expression in primate cells of cloned DNA coding for the hemagglutinin surface glycoprotein of influenza virus. AB - We have used simian virus 40 (SV40) DNA as a vector for expression of functional activity of a cloned influenza viral DNA segment in primate cells. Cloned full length DNA sequences coding for the hemagglutinin of influenza A virus (Udorn/72/[H3N2]) were inserted into the late region of a viable deletion mutant of SV40, and the hybrid DNA was propagated in the presence of an early SV40 mutant (tsA28) helper. Infection of primate cells with the hybrid virus produced a polypeptide similar in molecular size to the hemagglutinin of influenza virus, as shown by immunoprecipitation and gel electrophoresis. The polypeptide was glycosylated, as shown by incorporation of radioactive sugars. The putative hemagglutinin exhibited functional activity, as shown by agglutination of erythrocytes. In addition, an indirect immunofluorescence assay showed that the hemagglutinin polypeptide of the hybrid virus could be detected on the surface of infected cells. PMID- 6272306 TI - Knotted DNA from bacteriophage capsids. AB - The majority of the DNA prepared from tailless capsids of bacteriophage P2 by the phenol extraction procedure consists of monomeric rings that have their cohesive ends joined. Electron microscopic and ultracentrifugal studies indicate that these molecules have a complex structure that is topologically knotted; they have a more compact appearance and a higher sedimentation coefficient when compared with regular nicked P2 DNA rings. Linearization of these rings by thermal dissociation or repair of the cohesive ends by DNA polymerase I in the presence of all four deoxynucleoside triphosphates gives molecules that are indistinguishable from normal P2 DNA that has been similarly treated. The knotted nature of the majority of P2 head DNA is further supported by analyzing the products when these molecules are treated with ligase and the ligase-treated molecules are subsequently nicked randomly with DNase I. The data are consistent with the notion that, if such a molecule is first converted to a form that contains only one single-chain scission per molecule, strand separation gives a linear strand and a highly knotted single-stranded ring. The results suggest that the DNA packaged in tailless P2 capsids is arranged in a way that leads to the formation of a complex knot when the ends join. In an intact phage particle, the anchoring of one terminus of the DNA to the head-proximal end of the tail [Chattoraj, D. K. & Inman, R. B. (1974) J. Mol. Biol. 87, 11-22] presumably diminishes or prevents this kind of joining. The novel knotted DNA can be used to assay type II DNA topoisomerases that break and rejoin DNA in a double-stranded fashion. PMID- 6272307 TI - Biosynthesis of membrane-derived oligosaccharides: a periplasmic phosphoglyceroltransferase. AB - Membrane-derived oligosaccharides (MDO) are glucose-containing constituents of the periplasmic space of Escherichia coli whose biosynthesis is closely linked to the metabolism of membrane phospholipids. A periplasmic enzyme has now been discovered that catalyzes the transfer of phosphoglycerol residues between species of MDO or to certain glucose-containing model substrates such as gentiobiose (6-O-beta-D-glucopyranosyl-D-glucose). The partially purified enzyme has an apparent molecular weight of about 56,000 in gel permeation chromatography, and has an absolute requirement for divalent cations, of which Mn2+ is most active. Although the transferase activity appears to be the physiological function of the enzyme, at low concentrations of acceptor, the enzyme (Enz) acts as a cyclase, with the liberation of cyclic 1(3),2 phosphoglycerol, suggesting the following mechanism: (formula, see text). PMID- 6272308 TI - A second pathway of leukotriene biosynthesis in porcine leukocytes. AB - Incubation of suspensions containing polymorphonuclear and eosinophilic leukocytes with arachidonic acid led to the formation of two pairs of diastereomeric 8,(15S)-dihydroxy-5,9,11,13-icosatetraenoic acids and two erythro 14,15-dihydroxy-5,8,10,12-icosatetraenoic acids. The structures were elucidated by ultraviolet spectroscopy and gas chromatography--mass spectrometric analysis of several derivatives of each compound, catalytic hydrogenation, oxidative ozonolysis with steric analysis of alcohols, and comparison to reference compounds prepared by chemical synthesis. Experiments with 18O2 and H218O indicated that in all six compounds the hydroxyl group at C-15 was derived from molecular oxygen. Two of the diastereomeric 8,15-dihydroxy acids incorporated H218O at C-8, while the other two 8,15-dihydroxy products (also diastereomers) and the 14,15-dihydroxy compounds (geometric isomers) incorporated 18O2 at C-8 and C-14, respectively, in addition to C-15. Two of the 8,15-dihydroxy acids are formed by reaction of water with an unstable allylic epoxide intermediate, (14S,15S)-oxido-5,8,10,12-icosatetraenoic acid; the two 14,15-dihydroxy acids are proposed to be formed by reaction of activated molecular oxygen with the same epoxide, which in turn originates via 15S oxygenation of arachidonic acid. PMID- 6272310 TI - Tropomyosin is decreased in transformed cells. AB - The steady-state level and synthesis of a pair of polypeptides of Mr 33,000 and 35,000 in chicken embryo fibroblasts (CEF) transformed by Rous sarcoma virus (RSV) are significantly decreased relative to normal CEF; however, the decrease is more pronounced in the case of the Mr 35,000 polypeptide. These polypeptides have been identified as the alpha and beta subunits of CEF tropomyosin by selective staining with tropomyosin antibody, two-dimensional gel electrophoresis, partial peptide analysis, and solubility properties. The decrease in tropomyosin is shown to be a transformation-specific phenomenon in that it does not occur after infection with a virus deleted in src sequences. Decreased synthesis of tropomyosin is also observed in quail cells transformed by MC29 (a retrovirus with a different onc gene than that in RSV) and also in chemically transformed quail cells. The decreased in tropomyosin is probably not a direct result of the disruption of the microfilament system in transformed cells because disruption of the microfilament system with trypsin or cytochalasin B in normal CEF does not lead to a decrease in tropomyosin synthesis. A decrease in tropomyosin in CEF after transformation may be a result of a pleiotropic effect that results in the transcriptional inactivation not only of the tropomyosin gene but also of the fibronectin and procollagen genes described by others. PMID- 6272309 TI - Construction of a general vector for efficient expression of mammalian proteins in bacteria: use of a synthetic ribosome binding site. AB - With the premise that mRNAs transcribed in Escherichia coli from cloned eukaryotic DNA inserts do not possess the necessary regulatory signals for recognition by prokaryotic ribosomes, we have constructed a general plasmid vector carrying a chemically synthesized prokaryotic ribosome binding site that will ensure the efficient expression of eukaryotic proteins in E. coli. In addition to the regulatory signals necessary for ribosome recognition, the synthetic segment contains, at one end, a Pst I cleavage site which will direct its insertion to pBR322 DNA and, at the other end, a HindIII site to facilitate attachment of the passenger eukaryotic gene. Using simian virus 40 (SV40) tumor (t) antigen as a model system, we have ligated the SV40 DNA fragment containing the entire t antigen gene in tandem with the synthetic ribosome binding site to pBR322 DNA at the Pst I site, which lies within the coding sequence of the beta lactamase gene. Initiation of transcription at the beta-lactamase promoter would produce a chimeric mRNA with the synthetic ribosome binding signals and the SV40 sequence flanked by beta-lactamase coding sequences. Utilization of the synthetic regulatory signals for initiation of translation is demonstrated by the efficient synthesis, in bacterial transformants, of authentic SV40 t antigen. Excision of the entire SV40 insert by HindIII from those clones that have retained intact HindIII sites at the junction between the ribosome binding site and the SV40 sequence would allow insertion of other heterologous DNAs by using HindIII linkers. The efficient expression of any DNA insert would require that the entire coding sequence be contiguous and that its termini be randomized by treatment with exonuclease III and nuclease S1 to vary the distance between the translational initiation codon and the synthetic ribosome binding site. PMID- 6272311 TI - Glucocorticoid-mediated inhibition of ornithine decarboxylyase activity in S49 lymphoma cells. AB - Incubation of wild-type S49 lymphoma cells with glucocorticoids, such as dexamethasone and hydrocortisone, inhibits the activity of ornithine decarboxylase (L-ornithine carboxylyase, EC 4.1.1.17), the rate-limiting enzyme in the pathway of polyamine biosynthesis. The kinetics of this inhibition are more rapid than the glucocorticoid-mediated growth arrest in the G1 phase of the cell cycle or in glucocorticoid-mediated cytolysis of these cells. The inhibition of ornithine decarboxylase activity by corticosteroids is specific for steroids of the glucocorticoid class. Results obtained with variant S49 cells having lesions in the pathways of glucocorticoid or cyclic AMP action indicate that cytoplasmic glucocorticoid receptors, as well as nuclear transfer of steroid- receptor complexes, are required for the inhibition of ornithine decarboxylase activity but that this inhibition does not require hormonal activation of adenylate cyclase or cyclic AMP-dependent protein kinase. Because glucocorticoid mediated inhibition of ornithine decarboxylase occurs when cellular protein synthesis has decreased less than 20%, this inhibition may represent a specific glucocorticoid-mediated deinduction of ornithine decarboxylase in S49 cells. Inhibition of ornithine decarboxylase activity may offer a useful marker for suppression of growth and cell cycle progression in these and other lymphoma cells. PMID- 6272312 TI - Interactions of low density lipoprotein receptors with coated pits on human fibroblasts: estimate of the forward rate constant and comparison with the diffusion limit. AB - We present a theoretical study of the interaction of low density lipoprotein (LDL) receptors with coated pits. From published experiments we estimate that the forward rate constant, k+, for the binding of a LDL receptor to a coated pit on a human fibroblast at 37 degrees C is greater than or equal to 3 X 10(-10) cm2/sec, and the mean time an LDL receptor spends on the cell surface before being captured by a coated pit, tc, is less than or equal to 1.8 min. If, when an LDL receptor enters, it remains within the coated pit until the coated pit pinches off to form a coated vesicle, then k+ = 3 X 10(-10) cm2/sec and tc = 1.8 min. We derive expressions for the diffusion limit of k+ and tc when particles (LDL receptors) diffuse in two dimensions until they hit and are absorbed by circular absorbers (coated pits) that have finite lifetimes. The absorbers appear and disappear at equal rates so that their concentration remains constant. We use these expressions to show that a diffusion limit of k+ = 3 X 10(-10) cm2/ sec would be obtained if D, the diffusion coefficient for an LDL receptor on a human fibroblast at 37 degrees C equaled 3.3 X 10(-11) cm2/sec. Because this value is in agreement with the experimentally determined value for D, we conclude that random insertion of LDL receptors into the plasma membrane, followed by pure diffusional motion of LDL receptors on the cell surface until they are irreversibly absorbed in coated pits, is consistent with experiment. PMID- 6272313 TI - Membrane modification differentially affects the binding of the lactogenic hormones human growth hormone and ovine prolactin. AB - Human growth hormone (hGH) and ovine prolactin (oPRL) are both lactogenic as defined by their ability to induce milk-protein synthesis in vitro in the presence of insulin and hydrocortisone. At physiological concentrations, both hGH and oPRL have similar dose-response curves in a mouse mammary gland organ culture system. Binding of 125I-labeled hGH (125I-hGH) to lactogenic receptors is competed by both hGH and oPRL, and the competition curves are nearly superimposable. Moreover, solubilized membrane proteins bound with either 125I hGH or 125I-labeled oPRL (125I-oPRL) show the same sedimentation pattern on sucrose gradients. However, methylation of membrane phospholipids in the presence of the methyl donor S-adenosyl-L-methionine only increases the binding of 125I hGH. Binding of either 125I-oPRL or 125I-labeled bovine growth hormone (125I-bGH) was unaffected. Addition of concanavalin A (Con A) to the membranes decreased binding of 125I-oPRL to the lactogenic site by 80%, whereas 125I-hGH binding was decreased by only 40%, with the binding of 125I-bGH unaffected. However, both hGH and oPRL-bound proteins bind to Con A-Sepharose columns to the same extent. These results suggest that although hGH and oPRL bind to the same lactogenic site with similar affinities and elicit similar biological responses, modification of membranes either by phospholipid methylation or by Con A differentially affects the binding of these two hormones. PMID- 6272314 TI - Crucial role of thyroid hormone in x-ray-induced neoplastic transformation in cell culture. AB - Incubation of mouse embryo fibroblasts (C3H/10T1/2) in media depleted of thyroid hormone for 1 week rendered the cells completely resistant to the transforming action of an x-ray dose, 4 grays, that yields transformation frequencies (no. foci per surviving cells) of approximately 10(-3) in media supplemented with triiodothyronine (T3) (1 nM). Studies on the timing of the additions or removal of the hormone indicate that T3 was maximally effective when added 12 hr before irradiation and that progression from the time of irradiation to the appearance of foci (6 weeks) was independent of the presence or absence of the hormone. The dependence of x-ray-induced transformation on the concentration of T3 in the medium was virtually the same as that for augmentation of Na+,K+-ATPase activity. The latter effect was used as a measure of T3 induction of protein synthesis. A further indication of the involvement of protein synthesis in the process is the abolition of T3- and x-ray-dependent transformation by cycloheximide at a concentration (100 ng/ml) that inhibits 50% of protein synthesis. We propose that thyroid hormone induces the synthesis of a host protein that is an obligatory participant in x-ray-mediated transformation. PMID- 6272315 TI - Targeted killing of cultured cells by receptor-dependent photosensitization. AB - This paper describes a method, designated "receptor-dependent photosensitization," by which the receptor-mediated endocytosis of low density lipoprotein (LDL) can be used to deliver a photosensitizing agent, pyrene, to cultured human and animal cells. The hydrophobic core of LDL is extracted and replaced with pyrene covalently coupled to cholesteryl oleate. This reconstituted LDL enters cells in significant amounts only when the cells express LDL receptors, resulting in the accumulation of pyrene cholesteryl oleate within lysosomes. Subsequent exposure of the cells to ultraviolet light leads to cell death. Cells killed by this technique include normal and simian virus 40 transformed human fibroblasts, human A-431 epidermal carcinoma cells, Chinese hamster ovary cells, and mouse L cells, all of which express LDL receptors. Mutant fibroblasts from a patient with homozygous familial hypercholesterolemia, which lack LDL receptors, do not take up significant amounts of the pyrene containing LDL and are not killed by subsequent exposure to light. The current experiments establish the feasibility of receptor-dependent photosensitization as an efficient and selective method for killing cultured human and animal cells. PMID- 6272316 TI - Human nucleolus organizers on nonhomologous chromosomes can share the same ribosomal gene variants. AB - The distributions of three human ribosomal gene polymorphisms among individual chromosomes containing nucleolus organizers were analyzed by using mouse--human hybrid cells. Different nucleolus organizers can contain the same variant, suggesting the occurrence of genetic exchanges among ribosomal gene clusters on nonhomologous chromosomes. Such exchanges appear to occur less frequently in mice. This difference is discussed in terms of the nucleolar organization and chromosomal location of ribosomal gene clusters in humans and mice. PMID- 6272317 TI - Polymorphic DNA region adjacent to the 5' end of the human insulin gene. AB - The length of a segment of DNA associated with the human insulin gene, which has been localized to the short arm of chromosome 11, is heterozygous in 63% of 52 individuals analyzed. This polymorphic region is approximately 500 base pairs from the nucleotide encoding the 5' end of insulin mRNA. The polymorphism appears to be due to an insertion or deletion of DNA sequences so that DNA fragments of different length are generated when DNA from a heterozygous individual is digested with selected restriction endonucleases. PMID- 6272318 TI - Ethnic variation in Hpa 1 endonuclease cleavage patterns of human mitochondrial DNA. AB - The mtDNAs of 235 individuals from five ethnic groups were analyzed for restriction site variation by digestion with restriction endonuclease Hpa I, Southern transfer, and hybridization with 32P-labeled human mtDNA. Six different cleavage patterns (morphs) were found, all of which could be related to each other by single nucleotide substitutions. Differences were found in the frequency of these morphs among the populations. The largest difference observed was in the frequency of the morph most common in Caucasians and Orientals compared to the frequency of that found in Africans. This difference apparently originated by the sequence change G-T-C-A-A-C to G-T-T-A-A-C. This alteration permitted recognition by Hpa I but did not alter the amino acid sequence. Two other observed differences were due to separate substitutions occurring in the ribosomal RNA genes. Comparison with primate data shows that the morph with two fragments, found in 12.5% of Oriental and 4% of Bantu samples, might be the ancestral type common to all hominoids. These two conserved sites were localized in tRNA genes in the anticodon loop. Assuming that the two-fragment morph is ancestral, this finding is consistent with previous data suggesting that Asia is genetically central to the radiations that are thought to have given rise to the human ethnic groups. PMID- 6272319 TI - Genetic and molecular diversity in nondeletion Hb H disease. AB - Restriction endonuclease mapping of nondeletion alpha-thalassemia determinants from a variety of racial groups showed no detectable abnormalities within a 40 kilobase region of the zeta-alpha globin gene cluster. By using a zeta-specific probe, we defined three different types of interactions that give rise to Hb H disease, each involving a nondeletion alpha-thalassemia haplotype. mRNA analysis showed further diversity within these groups, indicating that there are at least three nondeletion determinations. PMID- 6272321 TI - Pleiotropic expression of Epstein--Barr virus DNA in human epithelial cells. AB - We have attempted to establish a system that can be used to study the association of Epstein--Barr virus (EBV) with epithelial cells. Attempts were made to transfect human carcinoma cells with EBV DNA. Successful transfection was confirmed by the expression of EBV-specific early antigen (EA), virus capsid antigen, and the presence of virus DNA. The transfecting preparation contained a mixture of EBV and cellular DNA extracted from two producer cell lines, P3HR-1 and AG-876. Our data suggest that virus DNA obtained from the P3HR-1 nontransforming, EA-inducing strain of EBV was lytically expressed in the epithelial tumor cells. The DNA derived from AG-876 cells, which produce a transforming, non-EA-inducing strain of EBV, also produced a lytic infection. PMID- 6272320 TI - Cellular homologue (c-src) of the transforming gene of Rous sarcoma virus: isolation, mapping, and transcriptional analysis of c-src and flanking regions. AB - The tumorigenic properties of Rous sarcoma virus are attributable to a 60,000 dalton protein, pp60v--src, encoded by a single viral gene, v-src. A homologous gene, c-src, that contains the information for a 60,000-dalton protein, pp60c- src, has been identified in all tested vertebrate cells. By screening a recombinant DNA library representative of the chicken genome, we isolated two overlapping DNA fragments that contain more than 30 kilobases (kb) of DNA spanning the coding sequences for pp60c--src. This 30-kb region is devoid of moderately or highly repeated sequences and shares homology with the entire viral gene and noncoding sequences 5' of v-src. Although v-src has an uninterrupted coding sequence, c-src is interrupted by a minimum of seven intervening sequences. At least two polyadenylylated RNAs seem to be encoded within the DNA we have isolated. The larger RNA, approximately 3.9 kb, is the presumptive c-src mRNA; the smaller transcript of about 2 kb hybridizes to DNA sequences several kb from the 3' end of the v-src/c-src homology area. PMID- 6272322 TI - Phencyclidine (angel dust)/sigma "opiate" receptor: visualization by tritium sensitive film. AB - [3H]Phencyclidine ([3H]PCP) binds specifically to an apparently single class of binding sites on slide-mounted sections of rat olfactory bulb (Kd = 46 nM; Bmax = 10.5 fmol per slice). Bound [3H]PCP can be displaced by nonradioactive PCP and a series of its analogs with relative potencies that correlate closely (P less than 0.001) with values determined in a rat discrimination test that utilized PCP as a cue. Although morphine, naloxone, and opiate peptides do not displace bound [3H]PCP, psychotomimetic benzomorphans, classed as "sigma opiates," are quite potent displacers in vitro and have PCP-like behavioral properties in vivo. These results suggest that phencyclidine and the sigma opiates act at the same sites. [3H]PCP binding sites were visualized by using tritium-sensitive LKB film analyzed by computerized densitometry and color coding. The [3H]PCP binds most densely to cortical areas, diffusely in neocortex, and somewhat heterogeneously in the laminae of the hippocampal formation and dentate gyrus. Most of the brainstem and spinal cord show low specific [3H]PCP binding, with gray matter generally showing more binding than white. PMID- 6272323 TI - Thiamine as an integral component of brain synaptosomal membranes. AB - Synaptic plasma membranes were prepared from rat cerebral cortex to determine if thiamine was localized in the membranes. The synaptosomes, prepared by discontinuous sucrose gradient centrifugation, were subjected to osmotic shock at pH 9.5 for 10--15 min and subfractionated on a discontinuous sucrose gradient. The two membrane fractions that were obtained were free of contamination by mitochondrial membrane and soluble fractions. In order to ensure specificity, thiamine was assayed fluorometrically before and after the addition of thiaminase I (thiamin:base 2-methyl-4-aminopyrimidine-5-methenyltransferase, EC 2.5.1.2). The thiamine content of the two membrane fractions was 9 and 10 pmol/mg protein. PMID- 6272324 TI - Properties of the gastric proton pump in unstimulated permeable gastric glands. AB - It was found that digitonin-permeabilized resting gastric glands retained considerable acid-secretory ability. Oligomycin abolished this, and ATP was able to bypass this inhibition and restore acid secretion. Moreover, the effect of anoxia was also bypassed by ATP in these preparations. As in intact glands, acid secretion was K+ dependent, and the concentration for half-maximal effect was 18.5 +/- 1.76 mM in Na+-free solutions, a value similar to that found for resting intact glands. The slight inhibition of ATP-dependent secretion by either valinomycin or 2,4-dinitrophenol, but total inhibition by a combination of the ionophores, is interpreted to mean that, in resting gastric glands, the in situ proton pump is electroneutral and the KCl pathway supplying K+ to the luminal face of the pump is probably electroneutral. PMID- 6272325 TI - Seventh Boyd Orr Memorial Lecture. Managing the mineral cycle. PMID- 6272326 TI - Food protein sources: implications for nutrient requirements. PMID- 6272327 TI - GABA receptors. PMID- 6272328 TI - The distribution of some identified peptide-containing pathways in the central nervous system including the basal ganglia. PMID- 6272329 TI - The use and abuse of clonidine as a psychopharmacological tool. AB - Because early reports on the mechanisms of action of clonidine emphasized its "pure" postsynaptic alpha-norepinephrine stimulatory properties. clonidine has enjoyed an ever-growing popularity in psychoneuroendocrinology research as a central NE agonist. Clonidine appears to have some psychotropic activity. However, its biphasic and sometimes unpredictable effects (such as the ones we obtained in our laboratory) and the evidence of activity at presynaptic receptors which interfers with the NE feedback loop and turnover indicates that the rational for its use in research is in need of complete reevaluation and may be even opposite to the one heretofore utilized. PMID- 6272330 TI - After-effect of microneurography in humans. Part IV. AB - Twenty subjects were followed up to three months after microneurographic recording of sympathetic nerve fiber activity to determine the nature and incidence of sequelae to the invasive microneurographic procedure. Skin or muscle parethesias lasting two weeks on the average were reported by the majority of subjects. Nine percent of the subjects also reported mild muscle weakness lasting two to four weeks. One subject reported skin paresthesias lasting up to three months postrecording. There was no incidence of permanent nerve injury. PMID- 6272331 TI - The wavelength dependence of the effect of 8-methoxypsoralen plus ultraviolet radiation on the induction of latent simian virus 40 from a mammalian cell. PMID- 6272332 TI - Predicted secondary structure of horse muscle acylphosphatase. Comparison with circular dichroism measurements. AB - We have predicted the secondary structure of horse muscle acylphosphatase by the statistical method of Chou and Fasman. In addition, we have studied the circular dichroism spectra of the enzyme, obtaining values for comparison to the predicted results. Discrepancies were found for the alpha-helix content estimated by the two methods. PMID- 6272333 TI - Inhibition of protein synthesis in cell cultures by vanadate and in brain homogenates of rats fed vanadate. AB - Following the observation of polysome disaggregation by vanadate, when Neuro-2a cells were incubated with vanadate, incorporation of 3H-leucine into protein was markedly inhibited. The inhibition of protein synthesis was dependent on vanadate concentration and on time of incubation. Inhibition of cell growth was also observed. Simultaneous measurements on vanadate-treated cells showed decreased Na,K-ATPase activity. Rats given sodium vanadate as their sole drinking fluid also showed an inhibition of brain protein synthesis (18 and 20% after 30 and 60 days, respectively, of treatment). Possible implications of the inhibition of Na,K-ATPase and of protein synthesis by vanadate are discussed. PMID- 6272334 TI - Behavioral response of the genetically obese (ob/ob) mouse to heat stress: effects of naloxone and prior exposure to immobilization stress. PMID- 6272335 TI - Effect of repeated fasting on ATPase activities in the brain of rats of different ages. PMID- 6272336 TI - Biochemical compartmentation of fish tissues. Distribution of gluconeogenic enzymes in the brain. AB - 1. Specific glucose-6-phosphatase and fructose-1,6-diphosphatase activity were found to be biochemically compartmentalized in four parts of the brain in nine nutritionally important fishes. 2. Glucose-6-phosphatase and fructose-1,6 diphosphatase activity were highest in the cerebrum and lowest in the cerebellum. 3. Piscivorous fishes had the highest gluconeogenic enzyme content, followed by catfishes and major carps. 4. After the liver and muscles, the various parts of the brain play an important role in carbohydrate metabolism. 5. A direct relationship between the stage of evolution and elevation of gluconeogenic enzyme levels was observed. 6. It is evident from the results and the discussion that evolution modifies the biochemical organization of fishes in general and of their brain in particular. PMID- 6272337 TI - Molecular cloning and mapping of a deletion derivative of the plasmid Rts 1. PMID- 6272338 TI - The functional organization of the octopine Agrobacterium tumefaciens plasmid pTiB6s3. PMID- 6272339 TI - Restriction endonuclease mapping of the octopine tumor-inducing plasmid pTiAch5 of Agrobacterium tumefaciens. PMID- 6272340 TI - A postauricular flap based on the contralateral superficial temporal vessels. PMID- 6272341 TI - Effect of lymphangiography on lymphedema. AB - Lymphangiography, using oily radiopaque Lipiodol Ultra Fluid has been shown to increase lymphedema in one-third of the patients with obstructive lymphedema and to cause lymphatic obliteration (demonstrated histologically). No effective element has been observed, although subclinical infection could not be ruled out. Allergy is another possible contributor, but the evidence from this series suggests that Lipiodol may act as a direct irritant when it is not rapidly cleared, as in the case of obstructive lymphedema, and proceed to obliteration of the residual lymphatics. On the basis of these findings, elective routine preoperative lymphangiography for lymphedema is considered strongly inadvisable. PMID- 6272342 TI - Effects of chronic delta 9-tetrahydrocannabinol administration on schedule controlled behavior of pigeons: cross-tolerance to pentobarbital and barbital. AB - Pigeons responding under a variable-interval (VI) 75-s schedule of food presentation were used to study cross-tolerance from delta 9-tetrahydrocannabinol (delta 9-THC) to pentobarbital and barbital. After initial dose-effect functions for pentobarbital and barbital were determined, the birds received delta 9-THC injections for 6 weeks. This chronic administration regimen resulted in a greater than 100-fold tolerance to delta 9-THC. Redetermination of the pentobarbital and barbital dose-effect functions during the chronic delta 9-THC regimen revealed statistically significant shifts to the right for the pentobarbital (0.191 log unit) and barbital (0.078 log unit) dose-effect curves. All six birds showed tolerance to pentobarbital, while four of the six showed tolerance to barbital. Blood barbital levels before and after chronic delta 9-THC was more prolonged and of much greater magnitude than the cross-tolerance to pentobarbital or barbital. The results demonstrate that cross-tolerance can develop from delta 9-THC to a barbiturate that normally undergoes little metabolism. PMID- 6272343 TI - Radiation-induced free radicals in oriented bromouracil-substituted DNA. PMID- 6272344 TI - ESR of irradiated 2-thiouracil single crystals. A 3alpha-hydrogen radical. PMID- 6272345 TI - Enhancement of antibody-complement cytotoxicity against virus-transformed hamster PARA-7 cells treated with heat and microwave radiation. PMID- 6272346 TI - Influence of whole-body gamma irradiation upon rat liver mitochondrial fractions. PMID- 6272347 TI - The Arrhenius plot behavior of a gamma-radiation-releasable, membrane-bound exonuclease. PMID- 6272348 TI - [Changes of leukocyte and lymphocyte values and of cytotoxicity in tumour carrying rats in dependence on a local irradiation and increase of cAMP level, respectively (author's transl)]. PMID- 6272349 TI - Computed tomography of the hepatobiliary system in infancy and childhood. PMID- 6272350 TI - Computed tomography of the kidney in infancy and childhood. PMID- 6272351 TI - Diagnostic imaging of pediatric abdominal masses: an overview. PMID- 6272352 TI - Abnormal fat thickness in newborn with nesidioblastosis. PMID- 6272353 TI - Lobar attenuation difference of the liver on computed tomography. AB - Lobar attenuation difference of the liver was demonstrated by computed tomography (CT) in 16 cases among over 1,000 cases in which CT scans of the liver were obtained. Cases were divided into two groups: with liver tumor and without liver tumor. Among the former group, hepatocellular carcinomas is the predominant lesion; tumor thrombus in the portal vein is presumably the cause of the attenuation difference. The latter group includes the patients with hepatocellular disease and clinically normal patients. In the former group, the tumor itself could not be noted in precontrast scans in eight out of 12 cases. When lobar attenuation difference of the liver is noted, a postcontrast scan should be obtained to determined whether there is a liver tumor in the lobe of lower attenuation. PMID- 6272354 TI - Computed tomography of localized dilatation of the intrahepatic bile ducts. AB - Twenty-nine patients showed localized dilatation of the intrahepatic bile ducts on computed tomography, usually unaccompanied by jaundice. Congenital dilatation was diagnosed when associated with a choledochal cyst, while cholangiographic contrast material was helpful in differentiating such dilatation from a simple cyst cy showing its communication with the biliary tract when no choledochal cyst was present. Obstructive dilatation was associated with intrahepatic calculi in 4 cases, hepatoma in 9, cholangioma in 5, metastatic tumor in 5, and polycystic disease in 2. Cholangioma and intrahepatic calculi had a greater tendency to accompany such localized dilatation; in 2 cases, the dilatation was the only clue to the underlying disorder. PMID- 6272355 TI - Hepatic ultrasonography in type I glycogen storage disease (von Gierke disease). Detection of hepatic adenoma and carcinoma. PMID- 6272356 TI - Radionuclide liver and bone scanning in the evaluation of patients with endometrial carcinoma. AB - Staging bone scans or skeletal surveys were obtained of 97 patients with endometrial carcinoma. Of the 77 patients with Stage I or II disease, no metastases were identified at staging. Three patients in the entire series demonstrated bony metastases; all of these metastases were detectable by radionuclide bone scan and radiographic bone survey. Eighty-nine patients were examined with radionuclide liver/spleen scanning at the time of staging. Four of the 89 initial scans were interpreted as demonstrating hepatocellular disease, and all four patients had abnormal liver function studies. Only one patient demonstrated a possible hepatic metastasis at initial diagnosis. This patient also had abnormal liver function studies. Based on these results, bone surveys and radionuclide bone scans are not indicated as screening procedures in endometrial carcinoma. It is suggested that screening for liver metastases in patients with endometrial carcinoma is not warranted in patients with normal liver function studies. PMID- 6272358 TI - Advances in clinical virology. PMID- 6272357 TI - Regulation of vascular tone, molecular mechanisms. PMID- 6272359 TI - Transfusion transmitted diseases. PMID- 6272360 TI - Marihuana. PMID- 6272361 TI - [Dopamine receptors (author's transl)]. PMID- 6272362 TI - [[Dopamine receptor; characterization of D-2 receptor in the intermediate lobe of the pituitary gland (author's transl)]. PMID- 6272363 TI - [Adrenergic receptors (author's transl)]. PMID- 6272364 TI - [Amine receptors and adenylate cyclase: on the role of GTP binding regulatory protein (author's transl)]. PMID- 6272365 TI - [Catecholamine(cyclic AMP) and Ca2+-calmodulin in the regulation of glycogen metabolism: an example for the dual control on the cellular function (author's transl)]. PMID- 6272366 TI - [Plasma catecholamine and the receptors in cardiovascular diseases (author's transl)]. PMID- 6272367 TI - [Polyamines and cancers (author's transl)]. PMID- 6272369 TI - Interaction between prostaglandins and cyclic AMP in the gastric mucosa. PMID- 6272368 TI - 15-Hydroxyprostaglandin dehydrogenase and delta 13 reductase content of gastrointestinal organs of rabbits and rats. AB - 15-Hydroxyprostaglandin dehydrogenase was measured in various gastrointestinal and non-gastrointestinal tissues from rabbits and rats. In addition, delta 13 reductase activity was measured in fundic and antral mucosae and gastric muscle from rabbits. In rabbits, antral mucosa contained the greatest activity of 15 hydroxyprostaglandin dehydrogenase and was 6 times more active than rabbit lung and fundic mucosa. In rat, duodenal mucosa was more active than antral mucosa or pancreas, suggesting that interspecies variations may exist. delta 13 reductase in rabbit gastric tissues was also more active in antral mucosa than in fundic mucosa or gastric muscle. The high activities of these enzymes in antral mucosa in conjunction with the known large prostaglandin content, particularly PGE2 and PGI2, suggest a large biological turnover of prostaglandin in these tissues. PMID- 6272370 TI - Influence of luminal acidification on bicarbonate transport by gastric and duodenal isolated mucosae. AB - Transport of HCO3- was measured by pH-stat titration in pairs of amphibian fundic or proximal duodenal mucosa using a modified Ussing chamber. Separate unbuffered solutions bathed the luminal sides of two tissues while their serosal surfaces were in contact with a common buffered solution. Lowering luminal pH bathing one fundic mucosa from 7.40 to 1.85 significantly increased HCO3- transport by another fundus. However, acidification of fundic mucosa did not affect duodenal HCO3- transport. In duodenal mucosal pairs, lowering pH from 7.40 to 5.46 caused an increase in HCO3- transport by the other tissue. Luminal H+ ion concentration may therefore regulate HCO3- transport via a humoral mechanism. PMID- 6272371 TI - Role of prostaglandins in the regulation of intestinal electrolyte transport. AB - The E prostaglandins (and to a lesser extent PGE2 alpha) stimulate active electrolyte secretion in mammalian small intestine and colon. They do so by stimulating intestinal mucosal adenylate cyclase and thereby increasing cAMP concentration. The diarrheagenic action of the prostaglandins is seen as a side effect of their therapeutic use, in certain hormone-secreting tumors, and in inflammatory lesions of the bowel in which leukocyte infiltrates are the probable sources of prostaglandin excess. Prostaglandins are also normally synthesized by intestinal epithelial cells and appear to play an important role in the physiologic regulation of intestinal fluid transport. In recent in vitro studies, we have shown that addition of arachidonic acid (K 1/2 congruent to 10(-6) M) also stimulates secretion, cAMP accumulation, and PGE2 production in rabbit ileal mucosa. In the continued presence of arachidonate, tachyphylaxis develops: both secretory and cAMP responses have a half-life of about twenty minutes and subsequent additions of arachidonate produce little or no further response. In contrast, PGE2 production continues undiminished. Similar tachyphylaxis develops when PGE2 itself is added. Resensitization following removal of PGE2 is rapid, 50% of the initial sensitivity being restored in 6-7 min. Prostaglandin desensitization has been noted in other cell systems and appears to be exerted on adenylate cyclase. PMID- 6272372 TI - PGE2 mediates the effect of pentagastrin on intestinal adenylate cyclase and Na-K ATPase activities. AB - The hypothesis that PGE2 mediates the effect of pentagastrin on jejunal Na-K ATPase and adenylate cyclase activities was tested in rats. Mucosal PGE2 and cAMP contents, Na-K-ATPase and adenylate cyclase activities were determined 45 min after IV pentagastrin (1 microgram/100 g b.w.) administration to pyloric ligated rats. Pentagastrin almost doubled mucosal PGE2 content as compared to that in saline-treated rats: 198 +/- 19 (S.E., N = 11) and 109 +/- 9 (S.E., N = 26) pg/mg tissue, respectively, inhibited mucosal Na-K-ATPase activity: 16.4 +/- 0.7 (S.E., N = 8) as compared to 26.7 +/- 4.0 (S.E., N = 13) mumole/mg protein/h in saline treated rats, stimulated adenylate cyclase activity by 146% and increased mucosal cAMP content by 80%. Pretreatment with indomethacin (4 mg/kg b.w., s.c./day x 2) prevented the increase in PGE2 content, the stimulation of adenylate cyclase activity and the inhibition of jejunal Na-K-ATPase activity induced by pentagastrin. The results reported thus suggest that the mechanisms whereby pentagastrin affects intestinal water and electrolyte transport are probably mediated by mucosal PGE2 and include inhibition of Na-K-ATPase activity and stimulation of the adenylate cyclase - cAMP system. PMID- 6272373 TI - [Metastasis of gastric cancer to the skin simulating actinomycosis]. PMID- 6272374 TI - [Studies on testosterone-binding proteins of skin supernatant of healthy women and those with acne]. PMID- 6272375 TI - A comparison of the ability of opioid peptides and opiates to affect active avoidance conditioning in rats. AB - Enkephalins reduce acquisition of an active avoidance response when administered intraperitoneally shortly before training. The present study examined whether microgram or delta opiate receptors are involved in this enkephalin effect. This was done by comparing the efficacy of micro- and delta-receptor agonists; by attempting to block the enkephalin effect with micro- and delta-receptor antagonists; and by comparing the characteristics of the effects of Met enkephalin and Leu-enkephalin. In addition, the efficacy of kappa-agonists in reducing acquisition was assessed. It was found that micro-agonists are inactive in this assay; several delta- and kappa-agonists are active. However, not all of the data are consistent with the adequacy of this receptor classification. The micro-receptor antagonist naloxone did not readily block the effect of Met- or Leu-enkephalin but neither did the micro/delta-antagonist, diprenorphine. An additional complexity is the emergence of differences in behavioral activity of Met- snd Leu-enkephalin. PMID- 6272376 TI - Cholecystokinin-octapeptide suppresses stress-induced eating by inducing hyperglycemia. AB - Intracerebroventricular administration of cholecystokinin-octapeptide (CCK-8) in unanesthetized, unrestrained rats suppressed stress-induced eating and produced hyperglycemia. Prior adrenalectomy markedly reduced the hyperglycemia effect of intraventricular CCK-8 and also decreased the suppressive effect of CCK-8 on stress-induced eating. Our data are compatible with the anorectic effect of CCK-8 being secondary to the hyperglycemia it produces. PMID- 6272377 TI - [Classical and ultrastructural neuropathology of aging processes in the human: a critical review (author's transl)]. AB - Although the primary interest in the course of studies on aging is focused on problems of function, morphology has an important role to play as a marker and as a control. Processes concomitant with brain aging are multiple and diverse. They ought to be correlated individually with functional changes. Statistical problems in population sampling for reference morphological studies are difficult. Bias is often a consequence of unstated underlying theories (Brain aging: a physiological process v. one of many pathological processes). Morphological criteria do not allow close application of animal models to aging problems in the human. The review considers the following lesions: 1. Brain shrinkage; 2. Neuronal loss; 3. Alterations in the outline of neurons; 4. Lipofuscin accumulation; 5. Corpora amylacea; 6. Senile plaques and amyloid deposits; 7. "Neurofibrillar degeneration"; 8. "Granulovacuolar degeneration"; 9. Hirano bodies. Vascular changes and their consequences are not considered. Lesions in the first group (1 to 5) are interpreted as non-specific and non-specifically correlated with aging; their correlation with functional loss is low. Lesion in the second group (6 to 9), on the contrary, are strongly correlated with each other and with dementia; they are interpreted as characteristic, not of aging itself, but of a widespread pathological condition: Alzheimer's disease sensu lato. A current line of research involves the hypothesis of a viral origin of this condition. PMID- 6272378 TI - Hormone-sensitive adenylate cyclase in glomerular cells: possible role for inflammatory diseases of the glomerulus. AB - Using the adenylate cyclase assay after Ross we examined hormone sensitivity of isolated glomerular cells. cAMP production was increased 1.3--1.6-fold by stimulation with isoproterenol, 1.5--1.8 times by prostaglandin E1 and 1.4--1.5 times by histamine. The isoproterenol reaction could be completely inhibited by propranolol, the histamine effect was abolished by the H2-blocking agent cimetidine. As control we applied sodium fluoride, which directly activates the catalytic adenylate cyclase unit, increasing the activity 1.8--2.7 times (depending on the method of homogenization). These findings could reflect some physiological or pathophysiological implications, which are discussed in the present report. PMID- 6272379 TI - Intracellular inhibition of catalase by alpha-methyldopa. AB - The effect of preincubation with alpha-methyldopa on catalase-dependent decomposition of H2O2 was studied in isolated rat hepatocytes. A direct assay of catalase function was obtained by measurement of O2 production under anaerobic conditions following additions of H2O2. Both the total O2 produced and the maximal O2 concentration attained were decreased by preincubation with alpha methyldopa. The results demonstrate that alpha-methyldopa-induced formation of Compound II is associated with loss of intracellular catalase activity. These results indicate that the hepatotoxicity of alpha-methyldopa may be related to an increased sensitivity to oxidative injury due to inhibited catalase function. PMID- 6272380 TI - Effect of certain neurotoxins and mixed function oxidase modifiers on glutathione S-transferase activity of rat brain. PMID- 6272381 TI - Cyclic nucleotides and cyclic nucleotide phosphodiesterases in kidneys from rats with experimental diabetes. AB - Experimental diabetes was produced in rats by administrations of streptozotocin (STZ) or alloxan (ALX). Some of the diabetic rats were started on daily insulin (NPH) therapy insufficient to control blood glucose. Rats were sacrificed one week or four weeks after confirmation of diabetes along with age-matched control rats. Analyses of cyclic nucleotide levels and of cyclic nucleotide phosphodiesterase activities in samples of kidney cortex revealed the following: cyclic AMP levels and activity of cyclic AMP phosphodiesterase were unaffected in all diabetic animals; cyclic GMP levels and cyclic GMP phosphodiesterase activity were unaffected in STZ-diabetic animals but were altered in ALX-diabetic animals. The data suggest that the altered cyclic GMP levels and degradation was due to a direct nephrotoxic action of ALX that is unrelated to the diabetic state. PMID- 6272383 TI - [Lafora disease]. PMID- 6272382 TI - Development of a total artificial middle ear prosthesis. PMID- 6272384 TI - [Herpes simplex meningoencephalitis]. PMID- 6272386 TI - [Catheterisation of the biliary tract and portal vein by Seldinger-technique following ultrasonically guided fine needle puncture (author's transl)]. PMID- 6272385 TI - Effects of beta-endorphin and morphine on the sleep-wakefulness behavior of cats. AB - The effects of beta-endorphin and of morphine SO4 (0.5 microgram and 2.0 microgram, respectively, injected intraventricularly) upon the sleep-wakefulness behavior of cats were examined. Both agents produced insomnia. Deep slow wave sleep was sharply inhibited, and rapid eye movement (REM) sleep was entirely suppressed. Light slow wave sleep, occurring in brief, isolated episodes, became the most abundant stage of sleep. The nuchal electromyogram was markedly increased after both agents. Naloxone (100 microgram/kg), injected subcutaneously 30 min before beta-endorphin or morphine SO4, entirely reversed these agents' effects on the two stages of slow wave sleep, and antagonized the exaggerated electromyogram. But naloxone did not counteract the REM-suppressant effect of either beta-endorphin or morphine SO4. Total sleep time reverted towards control values after naloxone pretreatment, but not entirely; the difference may be due to the persistent deficit of REM sleep. The data may indicate an involvement of an inner opioid in the regulation of sleep and wakefulness in the cat, and may point to a role for more than one endorphin receptor in the effects of opioids on the states of vigilance in cats. PMID- 6272387 TI - [Virus neutralizing antibody levels following acute IBR]. PMID- 6272388 TI - Studies on the transmission of human hepatitis A virus to stump-tailed monkey. AB - The newly-caught stump-tailed monkeys (Macaca speciosa) with negative antibody to hepatitis A were inoculated with human hepatitis A virus. The following findings were observed in the monkeys after inoculation: (i) the elevation of activities of the serum glutamic-pyruvic transaminase, lactate dehydrogenase and its isoenzyme (LDH5), (ii) the seroconversion of antibody to hepatitis A virus. (iii) the shedding of hepatitis A antigen in feces. These findings show that the stump tailed monkey (Macaca speciosa) is susceptible to infection of human hepatitis A virus. The virus recovered from the feces of the infected monkey, named as Hang zhou A-1A strain of hepatitis A virus, has experienced two generations of successful transmission in monkeys. PMID- 6272389 TI - Effect of intracerebral microinjection of naloxone on acupuncture- and morphine analgesia in the rabbit. AB - Although there are unequivocal evidences indicating the participation of endogenous opiate-like substances in acupuncture analgesia, their exact sites of action remain to be elucidated. From the results of localization studies by injecting minute amount of narcotic antagonist naloxone into discrete brain areas and assessing its effect on acupuncture analgesia in rabbits it is concluded that nuclei accumbens, amygdala, habenula and periaquaductal grey are the strategic sites for endogenous opioids to exert their analgesic effect. These brain areas are also of extreme importance for the realization of morphine analgesia. PMID- 6272390 TI - Assessing consequences of marine pollution by hydrocarbons using sponges as model organisms. AB - Pollution has been assessed as a mutagenic activity determined by the Ames test, using radiolabelled benzo[a]pyrene (BaP) as a model pollutant. Experimental animals were sponges, mainly Tethya lyncurium from the Northern Adriatic and from the Pacific near Catalina Island, California, U.S.A. Changes in ornithine decarboxylase (ODC) activity (ODC; EC 4.1.1.17) and polyamine concentrations with and without pollution were observed. There is a slow rise in ODC activity during the course of three-weeks exposure and a fast increase of polyamine levels during the course of one day. Mixed function oxygenase (MFO; EC 1.14) activity could not be detected in sponges. There was a significant concentration dependent coupling of radioactive BaP derivatives (BaPD) to the macromolecular fractions; the highest in protein, X 1000 greater than DNA and X 500 greater than RNA. Coupling is light-mediated and drops to zero in the dark. However when activated microsomal fractions from fish, that had been exposed to high level polycyclic aromatic hydrocarbon (PAH) pollution are added, dark incorporation rises to significant levels which can be decreased by the MFO inhibitor 7,8-benzoflavone (BP). The question of possible absence of DNA repair in the sponges and some implications are discussed. PMID- 6272391 TI - Cloned poliovirus complementary DNA is infectious in mammalian cells. AB - A complete, cloned complementary DNA copy of the RNA genome of poliovirus was constructed in the Pst I site of the bacterial plasmid pBR322. Cultured mammalian cells transfected with this hybrid plasmid produced infectious poliovirus. Cells transfected with a plasmid which lacked the first 115 bases of the poliovirus genome did not produce virus. PMID- 6272392 TI - Renin and angiotensin: the complete system within the neuroblastoma x glioma cell. AB - Cells of the homogeneous hybrid line neuroblastoma x glioma (NG108-15) have many neuronal properties. Immunocytochemical tests show that they contain both immunoreactive renin and angiotensin; direct radioimmunoassays show that they are positive for renin, angiotensin I, and angiotensin II; enzymatic assays show that they contain angiotensinogen and converting enzyme as well. The renin appears to be present in an enzymatically inactive form that can be activated by trypsin and then blocked by antiserum to purified mouse submaxillary renin. Renin concentration and activity are increased by enhancing cellular differentiation with dibutyryl cyclic adenosine monophosphate or by serum withdrawal. These findings demonstrate a complete renin-angiotensin system within these neuron-like cells, and suggest that activation of intracellular renin could generate angiotensin II. PMID- 6272393 TI - Both mu and delta opiate receptors exist on the same neuron. AB - Low concentrations of the relatively selective opiate receptor agonists dihydromorphine and normorphine (mu receptor agonists) and D-Ala 2-D-Leu 5 enkephalin (a delta receptor agonist) were applied to single enteric neurons while the frequency of action potential firing was recorded. Most neurons that were inhibited by the mu agonists were also inhibited by the delta agonist, but the two receptors could be distinguished by the higher concentration of naloxone required to antagonize the delta agonist. The results indicate that enteric neurons bear both mu and delta receptors and that cell firing is inhibited if either receptor type is activated. PMID- 6272394 TI - Quantitative autoradiography of [3H]muscimol binding in rat brain. AB - A simple quantitative autoradiographic technique for the study of neurotransmitter receptors that includes the use of a tritium-sensitive film permits saturation, kinetic, and competition studies of brain samples as small as 0.01 cubic millimeter. This technique was used to study [3H]muscimol binding in rat brain. Unilateral gamma-aminobutyric acid receptor supersensitivity was observed in the substantia nigra pars reticulata after production of localized lesions of the ipsilateral corpus striatum. PMID- 6272395 TI - Cloned viral protein vaccine for foot-and-mouth disease: responses in cattle and swine. AB - A DNA sequence coding for the immunogenic capsid protein VP3 of foot-and-mouth disease virus A12, prepared from the virion RNA, was ligated to a plasmid designed to express a chimeric protein from the Escherichia coli tryptophan promoter-operator system. When Escherichia coli transformed with this plasmid was grown in tryptophan-depleted media, approximately 17 percent of the total cellular protein was found to be an insoluble and stable chimeric protein. The purified chimeric protein competed equally on a molar basis with VP3 for specific antibodies to foot-and-mouth disease virus. When inoculated into six cattle and two swine, this protein elicited high levels of neutralizing antibody and protection against challenge with foot-and-mouth disease virus. PMID- 6272396 TI - Adenosine 3',5'-monophosphate modulates thyrotropin receptor clustering and thyrotropin activity in culture. AB - A biologically active rhodamine conjugate of thyrotropin binds at 4 degrees C to diffusely distributed membrane thyrotropin receptors which patch and become endocytosed into thyroid cells in a temperature-sensitive process. When the cells are first incubated with 8-bromo-cyclic adenosine monophosphate at 37 degrees C, the conjugate also binds to clustered receptors at 4 degrees C. Furthermore, 8 bromo-cyclic adenosine monophosphate reduces the amount of adenosine 3',5' monophosphate (cyclic AMP) induced by thyrotropin. Hence, increased intracellular cyclic AMP induces receptor patching and reduces the concentration of cyclic AMP normally induced by thyrotropin. This suggests that cyclic AMP acts both as the second messenger of thyrotropin and also as the regulator of the level of thyrotropin receptors. PMID- 6272397 TI - Integration and stable germ line transmission of genes injected into mouse pronuclei. AB - Genetic material has been successfully transferred into the genomes of newborn mice by injection of that material into pronuclei of fertilized eggs. Initial results indicated two patterns of processing the injected DNA: one in which the material was not integrated into the host genome, and another in which the injected genes became associated with high molecular weight DNA. These patterns are maintained through further development to adulthood. The evidence presented indicates the covalent association of injected DNA with host sequences, and transmission of such linked sequences in a Mendelian distribution to two succeeding generations of progeny. PMID- 6272398 TI - High levels of intracellular bombesin characterize human small-cell lung carcinoma. AB - "Small cells" or "oat cells" characterize a virulent form of lung cancer and share many biochemical properties with peptide-secreting neurones. The neuropeptide bombesin is present in all small-cell lines examined, but not in other lung cancer cell lines, suggesting that bombesinergic precursor cells in lung may give rise to this disease. PMID- 6272399 TI - [Mesenteric arteries. Anastomotic blood supply of the mesentery via the renal and adrenal, without hypertension (author's transl)]. AB - A rare clinical syndrome, observed during arterial stenosis of the large splanchnic trunks, Alfidi's syndrome associates digestive symptoms and hypertension. The latter results from reno-splanchnic shunts, reducing renal blood supply. In a patient aged 53 years, suffering for two months from severe epigastric pain with loss of weight of 10 kilos, arteriography showed anastomoses between the mesenteric circulation and the perirenal and adrenal vascular systems. However the latter were only slightly developed and consequently, insufficient to prevent infarction of the small intestine and colon. An anastomotic network neverthless existed, and renal arteriography demonstrated such shunts before the onset of hypertension which could have resulted from it. It is reasonable to believe that the renal steal syndrome did not yet have any effect on the blood pressure, as the anastomotic circulation towards the remaining digestive tract was still insufficient to ensure effective revascularisation. PMID- 6272400 TI - [Schwartz-Bartter syndrome during carcinoma of the pancreas (author's transl)]. AB - The authors report a case of carcinoma of the pancreas with inappropriate secretion of antidiuretic hormone in a 74 years old woman; the main static and dynamic characteristics of the Schwartz-Bartter syndrome are recalled together with the various therapeutic indications. Carcinoma of the pancreas remains exceptional among the numerous causes of Schwartz-Bartter syndrome. The relationships between carcinoma of the pancreas and pancreatitis are recalled in relation to this special case. PMID- 6272401 TI - [Adenocarcinoma of the small intestine. Endoscopic diagnosis (author's transl)]. AB - Adenocarcinomas of the small intestine are among the rarest tumours of the digestive tract. Their prognosis, which is usually very bad, is due to late diagnosis. In a 54 year-old-man, formerly tuberculous, the first sign was abdominal pain without any specific character. The diagnosis was confirmed by jejunal endoscopy, with biopsy under direct vision. The technique was made easier by the tumour developing on the first loop of small intestine, leading to a curative surgical operation. PMID- 6272402 TI - [Adriamycin for the treatment of non HOdgkin's lymphomas (author's transl)]. AB - One hundred and thirty one patients with non-Hodgkin's lymphomas (107 lymphomas and 24 histiocytic lymphomas) have been treated between January 1973 and January 1976, by an association of cyclophosphamide, vincristine and prednisone alone (CVP) or combined with adriamycin (CVP-A). All lymphomas stage I and II received CVP + radiotherapy: the actuarial analysis shows a stationary survival from 1 to 5 years at 89%. In the group of lymphoid lymphomas stage III and IV with nodular pattern, the survival is longer for patients who received CVP (69% at five years) than for patients treated with CVP-A (54% at three years). In the group of lymphoid lymphomas stage III and IV with diffuse pattern, the complete remission rate is 25% for the CVP and 67% for the CVP-A (p less than 0.01). However, the median duration of life is not different for the two groups (24 and 26 months respectively). In the group of histiocytic lymphomas, the complete remission rate in 50% for the CVP combination and 83% for the CPV-A. However, the median time of life which is 17 months for the CVP, is not reached for the CVP-A (Stationary survival at 90% from the 12th to the 36th months). These results show that the CVP combination is a good treatment for the lymphoid lymphoma with nodular pattern. The addition of adriamycin gives hope of long remissions and perhaps cure for histiocytic lymphomas, until now of bad prognosis. For the lymphoid lymphoma with diffuse pattern, the prognosis is still bad although we obtained with adriamycin a better rate of complete remission. The high dose chemotherapy rescued by autologous bone marrow transplantation may be a new possibility for this last type of non Hodgkin lymphomas. PMID- 6272403 TI - [Perforations of the colon and rectum during administration of barium enema. Six cases report (author's transl)]. AB - Perforations of the colon and rectum during administration of barium enema. Six cases report. The authors have stated six cases of accidental perforation occurring during barium enema examination. They recall their principal modality of unexpected arrival, in particular: the type of nozzle used, the means of injecting the contrasting liquid, the role played by the pathological associated colon-rectum lesions. The diagnosis may be postponed especially when the perforation is under the peritoneal seat. The pain during or after the opacification as well as the rectorragies are the most frequent alarming symptoms observed. The prognosis is on the whole serious but is can be improved by a rapid and appropriate therapy. PMID- 6272404 TI - [Infections due to anaerobic bacteria after abdominal surgery. Concerning thirty five cases (author's transl)]. PMID- 6272405 TI - [Histochemical observations of oral mucosa in the non-smoking cirrhotic patient (author's transl)]. AB - Buccal mucosa samples from non-smoking cirrhotic patients were studied with help of histochemical methods and compared with biopsies from smoking cirrhotic and healthy subjects. Very few differences were observed between the two cirrhotic groups, except for the following which were noted in the epithelium of the non smoking group: an increase of acid phosphatase, an increase of the DNA in the active cellular layers. The basal cells mitochondria were almost always reactive. The vascular walls showed enzymatic changes with a decrease of thiamine pyrophosphatase and alcaline phosphatase activity. These observations do not allow us to draw any conclusions regarding the pathogenesis of oral cancer in cirrhotic patients. PMID- 6272406 TI - [Mortality in appendectomy. About an homogeneous series of 5 348 cases (author's transl)]. AB - The authors have reviewed a series of 5 348 cases of appendectomy. They have registered 25 post-operative deaths, of which 22 were due to medical complications and 3 to surgical complications. Surgical complications, which involved a second operation in the early post-operative period, were noted in 31 patients. In over 50 p. cent of the cases, complications have been observed after appendectomies for common appendicitis. Most of the time, they were revealed by an occlusion, a peritoneal syndrome or a setting of intraperitoneal abscesses. In about 50 p. cent of the cases, the post-operative occlusion points to the presence of an abscess, a peritonitis or an hematoma. The most frequent and severe complications are caused by infection. In the present series, an overall mortality of appendectomy of 0.47 p. cent has been observed. In the patients who have to be reoperated, the overall mortality of 6.6 p. cent corresponds to a risk which is 15 times higher. PMID- 6272407 TI - [Tiapride and alcoholism (author's transl)]. AB - A new study of tiapride in the alcohol withdrawal syndrome is reported. Its purpose was to evaluate the efficacy and the lag-period of tiapride used systematically, on admission, by the intravenous route. Among twenty patients only one failure was recorded. In five patients, improvement only appeared slowly. It was never necessary to discontinue treatment because of side effects. The patients retained normal vigilance and were rapidly able to resume their activities. PMID- 6272408 TI - [Calculous stenosis of the duodenum. Five observations (author's transl)]. AB - Five observations of calculous stenosis of the duodenum after cholecysto-duodenal fistulization are reported. Four times, the pre-operative diagnosis has been made out by an opaque gastro-duodenal transit. The treatment is usually surgical, and its modalities are discussed about, but it is pointed out that the endoscopic duodenal desobstruction on patients running high operative risks presents a real interest. PMID- 6272409 TI - [Clinical and secretory results of the highly selective vagotomy. Study of sixty cases (author's transl)]. AB - The clinical and secretory status of sixty patients nine-fourty-four months after highly selective vagotomy for duodenal ulcer is exposed. Mortality and morbidity are very low. The long-term clinical results are Visick I and II in more than 85 p. cent of cases. The results of basal and peak acid output after insulin and pentagastrin are presented. The question: "is highly selective vagotomy sufficient for hypersecretors?" is discussed. PMID- 6272410 TI - [Septicemia due to an unusual micro-organism: Actinobacillus actinomycetum comitans (author's transl)]. AB - A new case of septicemia (without endocarditis) due to Actinobacillus actinomycetum comitans is described. The patient was a 59-year-old man with an intraventricular pacemaker. He was successfully treated by a combination of gentamicin and ampicillin. Human infections due to this demanding and slow growing micro-organism, which is not related to actinomycosis, are only exceptionally reported. In 73% of the cases the endocarditis is subacute. The study of the literature demonstrates the prevalence of male patients and the usual absence of leukocytosis. PMID- 6272411 TI - [Lack of correlation between frozen shoulder and HLA-B27 (author's transl)]. AB - HLA-B27 antigen is strongly correlated with ankylosing spondylitis and several other rheumatic diseases. However, data concerning frozen shoulder are contradictory. Bulgen reported an increased frequency of HLA-B27 which was not observed by Noy. We performed HLA typing of 46 French patients with frozen shoulder. We agree with Noy: frozen shoulder is not correlated with HLA-B27. PMID- 6272413 TI - [A new case of skin gangrene of the external genital organs in man (Fournier's syndrome). Review of the literature. Discussion of pathology and treatment (author's transl)]. PMID- 6272412 TI - [An usual observation of Bouveret syndrome (author's transl)]. AB - An unusual observation of Bouveret syndrome is reported. It occurred in a male patient, under the usual age for this disorder, with no history of cholelithiasis. Diagnosis was only established during the surgical procedure. There were neither radio-opaque stones nor air in the common biliary duct. Attempts to opacify the fistula failed. Gastro-entero-anastomosis, which is sometimes proposed because of the patient's poor general condition, is often necessary because of the local lesions. This procedure allows to wait until the local condition improves before undertaking the complete cure during the second procedure, with considerably lesser risk. PMID- 6272414 TI - [Immunologically agranulocytosis after ajmaline administration (author's transl)]. AB - The reversible ajmaline induced agranulocytosis is considered as the result of an immunologically mediated destruction. Striking results of leukocyte agglutination test has been observed in the reported case. They suggest that the granulocytic factor involved in this reaction could be in some cases, differentiation membranous antigens. These ones, located on the granulocytic precursors would be detected only during the granulocytic regeneration period. PMID- 6272415 TI - [Ileitis (probable Crohn's disease) and oral contraceptives (author's transl)]. PMID- 6272416 TI - [Concomitant pulmonary tuberculosis and bronchogenic carcinoma: a difficult diagnosis. Report of eighteen cases (author's transl)]. AB - The diagnosis in the same patient of the two diseases is very often delayed. Generally, tuberculosis is diagnosed first, and then cancer when the radiologic lesions extend despite of antituberculous treatment. In half of our cases, tuberculous lesions could not be individualized from cancer on the chest X-ray films. Antineoplastic chemotherapies may favour the development of tuberculosis, or atypical mycobacteriosis, in cancer patients, and it must be considered as an opportunistic infection in such patients. Clinicians should be aware of the possible coexistence of the two diseases and, in practice, ask for fiberoptic bronchoscopy and sputum cytology in tuberculous patients presenting atypical features, and for sputum examination in cancer patients. PMID- 6272417 TI - [Malignant bone tumors in children (author's transl)]. AB - Almost all malignant bone tumors in children are either osteogenic sarcomas or Ewing's sarcomas. Their origin and their site are different as well as the course of the disease and its treatment. Amputation or desarticulation in still the best local treatment for osteogenic sarcoma. Irradiation can benefit some patients. Conservative resection, followed by internal prosthesis, is still an experimental procedure. Results of chemotherapy combining high-dose methotrexate and adriamycin have been controversial. Prophylactic pulmonary irradiation seems to lessen the number of metastases. Controlled therapeutic trials are mandatory to assess the efficacy of these complementary treatments. Local cure is obtained in 90% of Ewing's sarcomas by high doses of radiotherapy but with a number of late sequellae. This tumor is also highly chemosensitive. Treatment of the infra clinical disease by intensive chemotherapy has significantly increased the number of long-term cures. Other therapeutic regimens combining chemotherapy, radiotherapy and limited conservative surgery are now under investigation in an effort to diminish the sequellae without decreasing the number of cures. PMID- 6272418 TI - [Frequency of complications and radiotherapeutic progresses in Hodgkin's disease (author's transl)]. AB - Two categories of iatrogenic effects are observed in Hodgkin's disease: the radiotherapeutic loco-regional complications; the systemic effects in which the long term carcinogenic risk is particularly important to consider. The first category can be avoided by the technical improvements which are steadily achieved. The second category can be minimized with a good therapeutic strategy in which the magnitude and intensity of the treatment are matched to the severity of each anatomical and clinical presentation. PMID- 6272419 TI - Current status of radionuclide scrotal imaging. AB - Scrotal imaging with technetium-99m sodium pertechnetate consists of a radionuclide angiogram and static scrotal scans. Utilization of this study in patients presenting with an acute scrotum can dramatically reduce the number of surgical explorations for acute epididymitis. It can also aid in other aspects of differential diagnosis in patients presenting with either an acutely enlarged and/or painful scrotum or a scrotal mass. Ambiguities in previous descriptions of perfusion through the spermatic and extraspermatic cord vessels are described and distinguished from scrotal perfusion. The clinical and scintigraphic spectrum of testicular torsion, including spontaneous detorsion, early acute testicular torsion, midphase testicular torsion, and late phase or "missed testicular torsion," is discussed and illustrated. The variety of patterns seen in acute epididymitis, including lateral and medial epididymal location, and focal epididymitis are described, as is the appearance of hydrocele as both a primary and secondary entity. The relationship of scrotal imaging to the overall clinical presentation and evaluation of these patients is emphasized in testicular torsion, torsion of the testicular appendages, epididymitis, abscess, trauma, tumor, spermatocele, and varicocele. The techniques, clinical utility, and relationship to radionuclide imaging of Doppler ultrasound and gray scale ultrasound scanning are reviewed. Doppler ultrasound results in many false negative studies in testicular torsion. Gray scale ultrasound is useful in clarifying the nature of scrotal masses. PMID- 6272420 TI - The value, limitations, and applications of nuclear dacryocystography. AB - Nuclear dacryocystography is simple, relatively harmless method of evaluating patients suspected of having abnormalities of the nasolacrimal drainage system. A group of normal saline containing approximately 100 muCi of 99mTc-pertechnetate is placed on the conjunctiva near the lateral canthus, and serial scintigrams are obtained as the pertechnetate flows along the tear strips, through the nasolacrimal drainage system, into the nasal fossa. By using a pinhole collimator with a very small aperture (1mm), the canaliculi, the nasolacrimal sac, and the nasolacrimal duct are readily visualized. When flow is impaired, the site of obstruction can often be identified. Contrast dacryocystography provides similar information but requires the injection of contrast material directly into a canaliculus. Nuclear dacryocystography provides good functional assessment of nasolacrimal drainage but has serious shortcomings in defining pathologic anatomy. Contrast dacryocystography outlines the anatomy well but often misses minor obstructions. The two studies are complementary and together provide an effective means of evaluating the nasolacrimal drainage system. PMID- 6272421 TI - Radionuclide salivary gland imaging. AB - Salivary gland imaging with 99mTc as pertechnetate provides functional information concerning trapping and excretion of the parotid and submandibular glands. Anatomic information gained often adds little to clinical evaluation. On the other hand, functional information may detect subclinical involvement, which correlates well with biopsy of the minor labial salivary glands. Salivary gland abnormalities in systemic disease such as sarcoidosis, rheumatoid arthritis, lupus erythematosus, and other collagenvascular disorders may be detected before they result in the clinical manifestaions of Sjogren's syndrome. Such glands, after initially demonstrating increased trapping in the acute phase, tend to have decreased trapping and failure to discharge pertechnetate in response to an appropriate physiologic stimulus. Increased uptake of gallium-67 citrate often accompanies these findings. Inflammatory parotitis can be suspected when increased perfusion is evident on radionuclide angiography with any agent. The ability of the salivary gland image to detect and categorize mass lesions, which result in focal areas of diminished activity such as tumors, cysts, and most other masses, is disappointing, while its ability to detect and categorize Warthin's tumor, which concentrates pertechnetate, is much more valuable, although not specific. PMID- 6272422 TI - Radionuclide imaging of nonneoplastic soft tissue disorders. PMID- 6272423 TI - Breast scintigraphy as an imaging modality in the diagnosis of breast masses. PMID- 6272424 TI - Increased scrotal activity. PMID- 6272425 TI - Radionuclide angiography: superior vena caval obstruction. PMID- 6272426 TI - [The response of fast-adapting units innervating the molar buccal gingiva of the cat mandible to ramp pressure and triangular vibratory stimuli (author's transl)]. PMID- 6272427 TI - Budd-Chiari syndrome and hepatic adenomas associated with oral contraceptives. A case report. PMID- 6272428 TI - Treatment of small cell carcinoma of the lung with methotrexate, doxorubicin, cyclophosphamide, and lomustine: a community-based study. AB - Forty-four patients with undifferentiated small cell carcinoma of the lung (SCCL) were diagnosed and treated at community hospitals. Patients with limited disease were treated with surgical resection or primary radiation therapy (RT) followed by chemotherapy; those with extensive disease received chemotherapy followed by RT if there was not a complete primary response. The chemotherapy used was a combination of methotrexate, doxorubicin, cyclophosphamide, and lomustine. Median survival for patients with both limited and extensive disease was 12 months, with a six-month survival of 89%. Half of the patients had recurrence in the lung. The toxicity was moderate and tolerable. We conclude that this combination chemotherapy plus radiation therapy carries acceptable toxicity and can be used in a community hospital to achieve response rates and survival of SCCL equivalent to that obtained in large cancer centers. PMID- 6272429 TI - Psycho-endocrine differences and correlations in symptomatic and asymptomatic climacteric women - the possible role of prolactin. AB - The hormonal and psychological profiles of 23 women with the climacteric syndrome were compared with those of 10 asymptomatic postmenopausal women. Statistical analysis of the results of this prospective study revealed significant differences in luteinizing hormone, oestradiol and prolactin values. Interesting correlations linking the clinical, hormonal and psychological parameters are presented and discussed. A hypothesis is advanced to explain the elevated levels of serum prolactin in asymptomatic postmenopausal women. A psycho-endocrine profile of the symptomatic climacteric women is presented. PMID- 6272430 TI - Artificial herd immunity to poliomyelitis in a semirural community in South Africa. AB - The sera of 139 children aged between 8 months and 6 years from the town of Port Alfred in the eastern Cape Province were examined for antibodies to the three types of poliovirus. A protective level of antibody to type 1 virus was found in 73.4% of the sera. The figures for types 2 and 3 were 94.2% and 68.3% respectively. The results indicate good herd immunity to poliomyelitis. No cases of paralysis or isolations of poliovirus had been reported from the area for the 6 years preceding the survey. The high degree of immunity is therefore unlikely to be due to circulating 'wild' virus to any great extent. It must be ascribed mainly to the immunity achieved by the administration of an average of 2.8 doses per child of trivalent live poliovirus vaccine. The low figure of 47.4% for immunity to all three types of virus could be due to enterovirus interference and inhibitory factors. A fourth dose of vaccine was thought to be indicated and is now given as a routine measure. Further investigations on the response to this will be undertaken. PMID- 6272431 TI - Alterations in morphine-induced analgesia and tolerance by various compounds. PMID- 6272433 TI - Failure of pinealectomy to affect acute changes in plasma levels of luteinizing hormone and prolactin in ovariectomized rats following delta 9 tetrahydrocannabinol administration. PMID- 6272432 TI - Role of dopaminergic-enkephalinergic interactions in the neurochemical effects of ethanol. PMID- 6272434 TI - Perinatal exposure to delta 9-THC in mice: altered enkephalin and norepinephrine sensitivity in vas deferens. PMID- 6272435 TI - Supersensitivity to microiontophoretically applied dopamine and GABA in feline spinal trigeminal and hypoglossal neurons following chronic haloperidol. PMID- 6272436 TI - Effect of acute and chronic ethanol treatment on gamma-aminobutyric acid levels and on aminooxyacetic acid-induced GABA accumulation. PMID- 6272437 TI - Na-K, adenosine triphosphatase activity in myelin, axoplasmic and synaptic plasma membranes isolated from rat brain: effect of ethanol. PMID- 6272438 TI - Mechanical performance of polyglycolic acid and polyglactin 910 synthetic absorbable sutures. AB - The mechanical performance of polyglycolic acid and polyglactin 910 sutures has been assessed using standardized reproducible tests. The results of these studies demonstrated distinct differences in their performance that can be related, in part, to their structural configuration. The size of the 4-0 polyglactin suture was significantly larger than that of the 4-0 polyglycolic acid suture. Since the strength of any suture is proportional to its cross sectional area, the strength of unknotted and knotted polyglactin 910 sutures was signficantly greater than that of the polyglycolic acid sutures. The strength of both sutures was similarly diminished by the formation of the knot. When these sutures were added to tissue, the breaking strength was even further reduced. The rate of decline in breaking strength of the two absorbable sutures in healing skin wounds was comparable. These absorbable sutures displayed knot security with a two throw square knot, 1 = 1. The ability of these sutures to reach knot break with this knot configuration is characteristic of sutures in which the surface exhibits a high coefficient of friction. Their rough surface also caused them to drag through tissue, making difficult to adjust tension on a continuous running suture. PMID- 6272439 TI - Redundant nerve roots of the cauda equina. AB - Four cases of redundant nerve roots of the cauda equina are reported, and the pertinent literature is reviewed. This disorder mainly affects males. The clinical history ranges from months to decades. The illness often starts with low back pain or sciatica, or both. Motor and sensory impairment of the legs dominate the further course of the disease. Serpentine filling defects in the column of contrast are a characteristic (but inconstant) feature on myelograms. Abatement of signs and symptoms occurs following adequate decompression of the redundant roots. PMID- 6272440 TI - [How can lower limb amputees best be integrated in the community? There is a great need for follow-up after discharge]. PMID- 6272441 TI - [The history of hepatitis B virus]. PMID- 6272442 TI - Effects of suloctidil on platelet survival time following cardiac valve replacement. AB - Platelet functions and blood clotting parameters were examined in 23 patients with a shortened platelet survival time after cardiac valve replacement. Treatment was given at random: for a first group antivitamin K with suloctidil and for the second group antivitamin K alone. Six weeks of treatment with antivitamin K alone did not induce any significant change in the platelet survival time, platelet retention or platelet factor 4 in plasma. In contrast, the shortened platelet survival time increased significantly after 6 weeks of treatment with suloctidil. Moreover the platelet retention and the amount of platelet factor 4 in plasma were significantly reduced, indicating that normalization of platelet survival is the result of decreased destruction, presumably at the level of the prosthetic surface. The clinical benefits expected from these results remain to be validated by a longer follow up period. PMID- 6272443 TI - Automated technique for measuring platelet adhesion to a surface in the presence of plasma. PMID- 6272444 TI - Characterization of a membrane-associated NADH-dependent cytochrome C reductase of human platelets. PMID- 6272445 TI - In vitro reactivity of alveolar macrophages and red blood cells with asbestos fibres treated with oxalic acid, sulfur dioxide and benzo-3,4-pyrene. AB - The effects of 3 UICC asbestos fibres (A chrysotile, crocidolite, amosite) were observed in vitro on red blood cells (RBC) and alveolar macrophages (AM). THe reactivity of the fibres after leaching with 0.1 N oxalic acid or adsorption of SO2 or benzo-3,4-pyrene (BP) was studied. The haemolytic activity of crocidolite and amosite was very low. A cytotoxic effect on AM occurred when the fibres were present in high concentration (100 microgram/ml), this was characterized by a release of both cytoplasmic (LDH) and lysosomal (beta-galactosidase) enzymes. The leached fibres were more haemolytic than the unleached ones, and more beta galactosidase (beta-Gal) than lactic dehydrogenase (LDH) was released from the AM. In contrast to the amphiboles, chrysotile fibres were highly haemolytic and induced a selective release of beta-Gal from AM. Leached fibres were less haemolytic and were cytotoxic for AM (both enzymes were released). Their in vitro reactivity was similar to that observed with quartz. The results showed that SO2 changed the reactivity very little. BP sorption on acid-leached chrysotile decreased the LDH release from AM. The difference in the in vitro reactivity related to the chemical state of asbestos fibres might explain the difference in their in vivo reactivity (latency, degree of fibrosis). This point is discussed. PMID- 6272446 TI - Changes in tissue cyclic AMP concentrations following an intravenous lethal dose of cholera enterotoxin in rabbits. PMID- 6272447 TI - Effect of phenolic compounds on in vivo activity of glucose-6-phosphatase in certain tissues of Notopterus notopterus. AB - The effects of phenol (P), dinitrophenol (DNP), pentachlorophenol (PCP), and their three combinations--(PCP + DNP)/P (highly antagonistic), (DNP + P)/PCP (additive) and (P + DNP)/PCP (highly synergistic)-- on glucose-6-phosphatase activity in liver, brain, kidney and gills of Notopterus notopterus were studied at three sub-acute concentrations (1/10, 1/15 and 1/20 the fraction of the 96-h LC50) after 15 and 30 days of exposure. When these chemicals were used separately, PCP exerted more effect than DNP and P. Maximum (71.56%) and highly significant (P less than 0.001) inhibition was observed in liver at the highest concentration of (P + DNP)/PCP combination after 30 days exposure, and lowest (3.64%) and insignificant inhibition in kidney at 1/20 of the (PCP + DNP)/P combination after 15 days exposure. Significant (P less than 0.05, P less than 0.01) stimulation in kidney or no effect in other tissues was observed in lower concentrations of P, DNP and (PCP + DNP)/P. PMID- 6272448 TI - Effect of procarbazine on benzodiazepine receptor in mouse brain. AB - Procarbazine, a non-benzodiazepine tranquilizer and anti-convulsant, significantly decreased the binding of [3H]diazepam to mouse cerebellar membranes. The drug treatment reduced both the affinity and density of the receptor. The effect, however, was more pronounced on the number of binding sites suggesting occupation of the receptor by procarbazine. The results suggest that benzodiazepine receptors may also be involved in the pharmacological action of non-benzodiazepine drugs. PMID- 6272449 TI - The coexistence of gastric cancer and duodenal ulcers. AB - The records of 356 gastric carcinoma patients who were treated surgically were reviewed. Among them, six patients (1.7%) had duodenal ulcers. Five cancers of the six patients were "early" gastric cancers classified as type "IIc" or "IIc + III" according to "The general rules for the gastric cancer study in surgery and pathology". The cancers of the six patients were located in the lower half of the stomach. Gastric secretion activity was normal in four and above normal in two cases. Three had signet ring cell carcinomas, two had poorly differentiated adenocarcinomas and one had a well differentiated adinocarcinoma. Our results were similar to other results reported in many papers. In Japan gastric carcinomas coexisting with duodenal ulcers were more often confirmed in the "early" gastric cancer stage. This is probably because endoscopic examinations of the stomach were more often performed in Japan than in other countries because of te large number of gastric disease patients. In addition, it is assumed that the growth of gastric cancer in the stomach is slow because of the very active gastric secretion. PMID- 6272450 TI - A young gastric carcinoma patient with umbilical and scrotal metastasis. AB - A 21-year-old gastric carcinoma patients who showed umbilical and scrotal metastasis is reported. Cutaneous metastases are seen in 3-4% of malignant tumors. The most frequent primary sites are the breast, lung, stomach, ovary, colon, uterus and kidney. Intraabdominal malignancies may produce umbilical metastasis. The primary cancer is usually in the stomach, colon, pancreas or ovary. Umbilical metastasis from the stomach seen in our case is known as Sister Joseph's nodule which is a sign of a poor prognosis. Metastatic scrotal carcinoma is a relatively rare condition which occurs by the hematogeneous pathway. However, in our case, direct invasion of tumor cells though the peritoneal cavity and along the lymphatic system of the spermatic cord was the most likely cause of scrotal metastasis. PMID- 6272451 TI - [On the problem of radiotherapeutic measures in addition to chemotherapeutic treatment of the microcellular bronchial carcinoma: analysis of recurrence rates (author's transl)]. AB - 83 patients with small-cell bronchial carcinomas were treated with a combination of cytostatic chemotherapy ("ACO II" = adriamycine, cyclophosphamide, vincristine) and irradiation of the mediastinum, the tumor umbra and the affected hilum with a focal dose of 30 Gy administered within a period of three to four weeks. During the same period, those patients presenting the favorable initial stage "limited disease" (LD) were additionally submitted to a prophylactic total brain irradiation with a dose of 30 Gy. 42 among 55 patients of stage LD had a complete remission. Among these 42 patients, fourteen had an intrathoracic recurrence, nine of them within the irradiation field. A cerebral recurrence after prophylactic brain irradiation was found in five of 42 cases. The conclusion is drawn that the irradiation dose which has to be administered into little intrathoracic fields must be increased. A dose of 45 Gy within 4 1/2 weeks is desirable. It seems that a prophylactic brain irradiation is only efficient in patients with a favorable prognosis (LD, CR). PMID- 6272452 TI - [Efficacy of protective effect of aminopropylamino-ethylthiophosphate in neutron beam, gamma or X-irradiation of mice (author's transl)]. AB - The efficacy of the radioprotective agent aminopropylamino-ethylthiophosphate (WR 2721) was tested against different kinds of radiation on female NMRI-mice. The animals were divided into 4 groups and were exposed to neutrons as main component, to secondary gamma rays in a tissue-equivalent phantom, to pure gamma rays and to X-rays respectively. The following dose reduction factors have been obtained: 1.05 for neutron-irradiation, 1.33 for gamma irradiation in tissue equivalent phantom, 1.69 for pure gamma rays and 2.08 for X-rays. The protective agent WR 2721 is apparently more efficient against low LET radiation. The mechanism of protection has been discussed in this respect. PMID- 6272453 TI - A method for the isolation of lipid droplet fractions from decapsulated rat adrenals. AB - Ultrastructural and cell fractionation studies implicate lipid droplets in the storage of cholesterol and in the secretion of steroids. To evaluate the role of the lipid droplet in steroidogenesis, a discontinuous gradient centrifugation method has been developed for the isolation of both lipid droplet and non-lipid fractions from decapsulated rat adrenal homogenates. Steroids were extracted from the fractions with chloroform/methanol; the cholesterol ester, cholesterol and corticosterone in each extract were purified using a single chromatogram and the purified steroid and sterols were assayed fluorometrically. The lipid droplet fraction contained 85% of the esterified cholesterol and 32% of the free cholesterol found in whole gland extracts. Although adrenal lipid droplet fractions isolated from non-stimulated control animals contained 65-79% of the total corticosterone assayed in extracts of the whole gland, in vivo injections of ACTH did not increase corticosterone in this fraction. On the other hand, the corticosterone measured in non-lipid fraction extracts increased significantly following ACTH treatment. These results suggest that the synthesis/release mechanism for corticosterone is not associated with the lipid droplets but may involve specific components in the non-lipid fraction. The function of lipid droplet corticosterone is unknown. PMID- 6272454 TI - Disparate recovery of resting and stimulated oxidative metabolism following transient ischemia. AB - To assess the residual effects of transient cerebral ischemia on mitochondrial oxidative metabolic function, changes in the reduction/oxidation state of cytochrome a,a3 and relative local blood volume were measured in situ from the exposed cerebral surface of rat brain before and after 10 minutes of carotid artery ligation. During the ischemic interval, cytochrome a,a3 became reduced and electrocortical activity was abolished. During the first 20 minutes of reperfusion cytochrome a,a3 was hyperoxidized beyond baseline with eventual recovery to the original steady state. Electrocortical activity returned more slowly. Increased energy demand induced by electrical stimulation of the cortex produced transient oxidation of cytochrome a,a3. The amplitude of this oxidative response was decreased during the first 30 minutes of reperfusion. During the first 2 hours of reperfusion the time required for re-reduction of the oxidative response was lengthened despite the recovery of baseline mitochondrial redox state. These data demonstrate residual metabolic dysfunction after transient ischemia not apparent under "resting" conditions but evident when the system is required to perform additional "work." We speculate this metabolic dysfunction could be due to relative substrate limitation. PMID- 6272455 TI - Thresholds in cerebral ischemia - the ischemic penumbra. PMID- 6272456 TI - Incomplete transient ischemia: a non-destructive evaluation of in vivo cerebral metabolism and hemodynamics in rat brain. AB - Differential near infrared spectrophotometry was used to monitor sequential in vivo alterations in cerebral hemoglobin saturation, blood volume, and cytochrome c oxidase reduction/oxidation responses during and after a period of incomplete transient ischemia (acute, reversible common carotid artery occlusion). In this study the rat brain was monitored non-invasively in the intact skull by transillumination. The data show that an increase in cerebral deoxygenation of hemoglobin, which occurs simultaneously with a decrease in blood volume subsequent to carotid ligation, acts as a compensatory mechanism to assist in maintaining aerobic energy metabolism. The observations also demonstrate that in this species the effects of bilateral carotid occlusion on the cerebrovascular parameters are not necessarily irrevocable. The intramitochondrial metabolic alterations, as evaluated by cytochrome c oxidase redox transitions, are reversible as long as the systemic arterial blood pressure does not fall below a value of approximately 40 mm Hg. These data suggest that possibility of being able to use a critical reduction level of cytochrome c oxidase as an early indication of ischemia-induced cerebral metabolic dysfunction prior to major changes in high energy stores. PMID- 6272457 TI - Recovery of o'nyong-nyong virus from Anopheles funestus in Western Kenya. AB - O'nyong-nyong (ONN) virus first appeared nearly 20 years ago and was responsible for one of the largest arbovirus outbreaks ever documented. Since the original outbreak ended, ONN activity, as determined serologically, gradually declined on the Kano Plain in western Kenya. In June, 1978, a virus similar or identical to ONN was isolated from a pool of Anopheles funestus Giles captured at Ahero on the Kano Plain. The possible implications of this isolation are discussed. PMID- 6272458 TI - Field diagnosis of sleeping sickness in the Ivory Coast. I. Comparison of the miniature anion-exchange/centrifugation technique with other protozoological methods. PMID- 6272460 TI - [Treatment of recurring chronic parotitis in children with intracanalicular injections of lipiodol]. PMID- 6272459 TI - Left-side pleuropulmonary amoebiasis: a case report from Chad. PMID- 6272461 TI - Control of ionic permeability by membrane charged groups: dependency on pH, depolarization, tetrodotoxin and procaine. AB - The membrane permeabilities of K, Na, and Cl were determined in crayfish giant axons from pH 3.8 to 11.4. In general, cation permeability increases with pH while anion permeability decreases. In normal saline (Ko = 5.4 mM, pH = 7), P(K) = 1.33 X 10-5, P(Cl) - 1.49 X 10-6, and P(Na) = 1.92 X 10-8 cm/s. Increasing external potassium results in a dramatic membrane conductance change around Ko = 12 mM (Vm = -60 mV) which results primarily from changes in P(Na) and P(Cl). In elevated potassium (Ko = 40 mM, pH = 7), P(K), P(Cl), and P(Na) increase by 1.45, 8.1 and 14.2. In the potassium depolarized axon, P(Na) and P(Cl) show a cooperative change when pH is altered through the imidazole pK region (pK = 6.3). These changes are not seen in normal saline, or with P(K). A Hill coefficient n = 4 was found for the cooperative change of P(Na) and P(Cl). An interpretation here is the four protein molecules interact to form the Na and Cl ionic channels. Tetrodotoxin has minimal effects on passive permeabilities but reduce the Hill coefficient n for P(Na) but not P(Cl), while procaine reduces n for both P(Na) and P(Cl). The results show that membrane fixed charged groups have varied association and control over the different ion permeabilities. In addition, membrane conformational changes are also involved in permeability control. PMID- 6272462 TI - [Acute kidney failure developing after the intravenous administration of urotrast]. PMID- 6272463 TI - The effect of autonomic drugs on ureteric peristalsis: a canine in vivo study. AB - An in vivo canine model was developed in which the renal pelvis was perfused by a cannula nephrostomy and ureteric activity assessed by monitoring bolus volume and interperistaltic interval. The effect of autonomic drugs showed that the ureter contained alpha-adrenergic receptors which on stimulation caused an increase in ureteric contraction rate and a decrease in bolus volume. With beta-adrenergic receptor stimulation, there was complete inhibition of ureteric peristalsis for a variable period and evidence is presented that the beta-adrenergic receptors may be beta 1 rather than beta 2. Cholinergic stimulation of ureteric rate was observed, but seemed to be mediated indirectly via alpha-adrenergic receptors. No significant change was seen in ureteric activity with adrenergic blocking agents alone, suggesting that the physiological importance of these receptors in normal activity is questionable. PMID- 6272464 TI - Ultrastructure of collagenase-isolated acini (alveoli) from lactating mammary tissue. PMID- 6272465 TI - [Abrikosov's granular cell tumor localized on the vocal fold]. PMID- 6272466 TI - [Surgical treatment of benign liver tumors]. PMID- 6272467 TI - [Cytogenetic study of bovine leukosis]. AB - Nine dairy cows aged four to eleven years were subjected to examination by chromosomal analysis. These cows had been found, by haematological examination, to suffer from leucosis. The study also covered one ten-day-old calf - heifer. The test group included two dam-daughter pairs. The animals belonged to the Black Pied Lowland breed. The blood was sampled from vena jugularis and the karyotypes were processed and evaluated by the method after Moorhead et al. (1960), modified by Lojda et al. (1974). A list was kept for each animal. The tested animals were included in classes by the percentages of the chromosome aberrations: class I - two animals (up to 10% of aberrations), class II - seven animals (from 10% to 20% of aberrations), class III - one animal (above 20% of aberrations). Hyposomy was found in all cases, polysomy and hyperploidy in four cases. Structural aberrations were observed in nine cases, breaks being the most frequent anomalies (7 cases). Breaks on sexual chromosomes were observed in five cases, including the dam-daughter pairs; centric fusion occurred in one case and mixed aberrations in two cases. PMID- 6272468 TI - [Growth of bovine fetal cells after intrauterine virus infection of the fetus]. PMID- 6272469 TI - Eye disease in IBR. PMID- 6272470 TI - Pilot study of intravenous ifosfamide plus oral acetylcysteine in the treatment of non-small cell lung cancer. PMID- 6272471 TI - Adsorption of solvents by activated charcoal, polymers, and mineral sorbents. PMID- 6272472 TI - [Use of the passive hemagglutination reaction to study Aujeszky's disease]. AB - Experiments were carried out with the use of the passive hemagglutination test to study Aujeszky's disease, the antigen employed being the avirulent MK-35 strain of the disease virus. It was established that the passive hemmagglutination reaction could be referred to as a specific and rapid method in the study and diagnosis of the disease. Results showed that the vaccinal MK-35 strain was successfully used as an antigen in the test. In the treatment with an MK-25 vaccine there were predominantly antibodies detectable with the passive hemagglutination reaction. PMID- 6272473 TI - [Comparative trial of the immunogenic properties of commercial foot-and-mouth virus strains]. AB - Comparative investigations on the antigenic relation and the immunogenic properties of the foot-and-mouth disease virus types O, A, and C. were carried out with the aid of the quantitative CFT and experiments with guinea pigs. It was established that the viruses of a given type were closely related antigenically and belonged to one and the same subtype group (R greater than of equal to 70 per cent). Immunogenicity studies revealed that strains O1Y A5f, C 564, and Cf were best in terms of immunogenic properties. It was also found that, generally, type O strains were weak in this respect, while all strains of type C tested had good immunogenic properties. PMID- 6272474 TI - [Creation of an avirulent mutant of Aujeszky's disease virus by exposure to 5 bromodeoxyuridine]. AB - A mutant with new biologic properties has been obtained at the cultivation of a virulent strain of the Aujeszky's disease virus in chick embryo fibroblast cultures, following a definite number of passages. It has proved resistant when culturing at the present of 5-brom-desoxyuridine and 5-iod-2-desoxyuridine, retaining its infections titer. It produces small plaques and destroys cell cultures of a tiny granular structure, the plaque-forming titer being 5. 6X10-7. The mutant has been shown to be avirulent for sheep, suckling pigs, and rabbits. In confers solid and lasting immunity in vaccinated animals which are virus-free and show no virus excretion. The newly obtained MK-35 mutant has proved to be stable in terms of its new biologic properties. PMID- 6272475 TI - [Determination of the leukemia virus in the lymphocytes of sick cattle]. AB - Experiments were carried out to establish the leukosis virus in lymphocytes from the peripheral blood of spontaneously infected cattle. The infected animals were detected by means of the agar gel immunodiffusion test in demonstrating the presence of leukosis virus antibodies. Lymphocytes were obtained from the blood of positively reacting animals, which were cultured at 37 degrees C for 3 to 5 days in a 'MEM' nutrient medium with the addition of 10 per cent fetal calf serum and antibiotics. The virus in the cell culture liquid was concentrated with ammonium sulphate and polyethylene glycol after which it was tested for antigenic properties. Parallel to this the lymphocytes were studied via the electron microscope prior to and after culturing. It was found that at culturing such lymphocytes the culture liquid contained the virus, which, after concentration exhibited antigenic properties in the agar gel immunodiffusion test, forming a precipitation line with a leukosis-positive serum. As found by the electron microscope there were in the lymphocytes prior to and after cultivation viral particles which proved similar to the leukotic 'C' type viruses with budding of the cell membrane that was characteristic of the bovine leukosis virus. PMID- 6272476 TI - [Use of cell agglutination with concanavalin A to detect cattle infected with the leukemia virus]. AB - The method of cell agglutination with concanavalin 'A' was employed to study lymphocytes from the peripheral blood of cattle spontaneously infected with the virus of leukosis. The infection in the animals was demonstrated through the agar gel immunodiffusion test based on the presence of antibodies. As controls served lymphocytes of normal cattle that were serologically and clinically negative for leukosis as well as lymphocytes, of cattle that were immunized with the virus of the mucous disease and with adeno-, rota, and parainfluenza-3 viruses. It was found that 10(6) lymphocytes of cattle infected with the leukosis virus was agglutinated by concanavalin 'A' used at the rate of 500-2000 microgram/cm3, while the same amount of lymphocytes of normal animals or animals that were immunized against the other viruses got agglutinated by concanavalin 'A' at 4000 8000 g/cm3. PMID- 6272477 TI - [Biological properties of the MK 35 mutant of Aujeszky's disease virus (Pseudorabies virus)]. AB - Clinical, virological, and morphological studies were carried out with a total of 15 twenty-day-old pigs and 55 rabbits of 2.5 kg body weight. All were experimentally infected with a new mutant, MK 35 (3.5 x 10(7) plaque forming units) of the pseudorabies virus. The changes taking place in the test animals were followed up in their dynamics from the first to the thirtieth day after injecting the virus. No clinical signs speaking of the development of Aujeszky-s disease were noted, neither was the virus isolated from the tonsils, lungs, lymph nodes, and the central nervous system of the treated animals. Histologically, there was in the central nervous system a slightly expressed nonsuppurative meningoencephalitis which was most clearly manifested on the fourth day after the animals were infected. Later on the changes appeared to be more slightly visible, and on the 8-10th day they were no longer seen. At the 48th hour following the inoculation and up to the end of the experiment eosinophil leukocytes were present in the tonsils, lymph nodes, and spleen of all infected animals as well as within the parenchyma of the adrenal glands of the pigs only. The presence of eosinophil leukocytes in the enumerated organs of the test animals is referred to as the morphologic manifestation of immunogenesis--the result of an enhanced defense status of the host induced by the MK 35 mutant of the pseudorabic virus. PMID- 6272479 TI - Serological investigations into the presence of antibodies to Yaba 1-Lednice 110 virus in Romania. PMID- 6272482 TI - Investigations on the circulation of viruses occurring in the respiratory tract of apparently healthy schoolchildren aged 7 to 14 years. AB - A number of 92 viral agents could be isolated from 1,000 nasopharyngeal secretion samples collected from apparently healthy schoolchildren aged 7-14 years. The following viruses were identified, in decreasing order of frequency: 42 influenza A/USSR 90/77 (H1N1), 20 adeno-, 20 parainfluenza, 8 Coxsackie B1 and 2 herpes virus strains. PMID- 6272483 TI - Cytogenetic modifications induced in human diploid cells by mixed infection with measles and cytomegalic virus. AB - Chromosome alterations induced in human diploid cells by separate or mixed infection with measles and cytomegalic virus were analyzed. Chromosome changes characteristic of each virus could be made evident in the case of the mixed infection. Of particular interest are the cytogenetic alterations associated with syncytium formation, both in separate measles virus infection and in the mixed one. The cytogenetic consequences of premature chromosome condensation, of the synchronism in division of polykaryocyte nuclei, of chromosome contraction and pyknosis are discussed. PMID- 6272484 TI - The effect of violamycin BI on different biological systems. Note I. The action of violamycin BI on cell cultures infected or not with herpes simplex virus type 1. AB - The effect of violamycin BI (VBI) was investigated in different cell substrates, infected or not with herpes simplex virus type 1 (HSV-1), strains VR3 and Rapp-1. The cytotoxic dose of VBI was established in calf kidney and human embryo cell cultures. Low drug concentrations induced moderate cytotoxic changes, allowing the study of the cells for 4-6 days. The action of VBI on HSV-1 was evident mostly in the early phases of the virus multiplication cycle. The antiviral effect of VBI was due both to the direct action of the drug on HSV and to its interaction with the nucleic acids of the host cells supporting HSV multiplication in their nuclei. PMID- 6272485 TI - Investigations on the presence of antibodies to papova viruses in patients with different forms of cancer and in other categories of patients or apparently healthy subjects. Note I. Neutralizing antibodies to SV-40 in the cerebrospinal fluid of patients with brain tumors and chronic neurological diseases. AB - Neutralizing antibodies to SV-40 were detected in cerebrospinal fluid samples from 16 out of 43 patients with brain tumors and in 6 out of 20 patients with different nonmalignant chronic neurological diseases. The patients' age excluded the correlation with a vaccine-associated or occupational contact with SV-40. The results ascertain the occurrence of SV-40 or of an antigenically related virus in the population of Romania. PMID- 6272486 TI - Investigations on the presence of antibodies to papova viruses in patients with different forms of cancer and in other categories of patients or apparently healthy subjects. Note II. Hemagglutination-inhibiting serum antibodies to BK virus. AB - Hemagglutination-inhibiting serum antibodies to BK virus (BKV were detected in patients with different forms of cancer and in blood donors (positivity percentages: 67.04 and 57.78, respectively). No such antibodies were found in a group of children 1 to 14 years of age. The data point to the presence of a latent human infection with BKV in the population of Romania. PMID- 6272487 TI - Investigations on the presence of antibodies to papova viruses in patients with different forms of cancer and in other categories of patients or apparently healthy subjects. Note III. Hemagglutination-inhibiting serum antibodies to polyoma virus. AB - Hemagglutination-inhibiting antibodies to polyoma virus could be detected in sera from apparently healthy subjects, patients with nonmalignant respiratory diseases and patients with different forms of cancer. The positivity percentages (12.97, 12.92 and 16.36, respectively) and the titers recorded were lower than in the case of BK virus. The results obtained suggest a slight antigenic relationship between polyoma and BK virus. PMID- 6272488 TI - [Malignancy of hormonally active ovarian tumors]. AB - 401 cases of hormone-producing tumors of the ovary were studied (190 theca-cell, 169--granulose cell, 25--mixed thecagranulosa cell and 17--virilising tumors). Histological diagnosis, clinical stage and end results of their treatment were compared. Theca-granulosa cell tumor is considered to be the most malignant form of these neoplasms. Pure theca-cell tumor proved to be benign in most cases. Of special interest is a high percentage of patients being cured at stage 1 (FIGO classification) of neoplastic process, that stands, in the authors' opinion, in close relationship with comparatively good results obtained in these patients. PMID- 6272489 TI - [Incorporation of influenza virus antigens into the envelope of the avian oncornavirus]. AB - The mixture of influenza virus and pseudotype of Rous sarcoma virus--RSV (PAB-A) grown in chick embryos or cell culture of chick fibroblasts produces a progeny which can be partially neutralized by a heterologous immune serum. The fraction of viral particles, the transformation activity of which is susceptible to suppression by both homologous anti-RSV (PAB-A) and heterologous serum, immune to influenza virus, amounts to approximately 20%. After passivation, these particles lose this property which suggests a non-genetical character of the phenomenon. Hence, dual infection results in phenotypic mixing and formation of particles with nucleoproteid of RSV (PAS-A) which contain antigens of avian sarcoma and influenza viruses in their envelopes. PMID- 6272490 TI - [Dynamics of the kinin system components of the blood plasma in acute lymphoblastic leukemia]. PMID- 6272491 TI - [Role of the host cell in virus persistence]. PMID- 6272492 TI - [Trial of polyguacyl safety and tolerance]. AB - The results of studies of the antiviral and interferon-inducing activity of the synthetic interferon inducer polyguacyl in white mice as well as the results of the study of safety and tolerance of this drug given to human subjects as aerosol and intranasally are presented. Both modes of administration to mice induced production of endogenous interferon, although after intranasal inoculation high interferon titres in the blood serum of the animals were observed for longer periods of time, whereas after aerosol administration interferon disappeared more rapidly. Significant antiviral protection was achieved only by the intranasal administration of the inducer resulting in 84.0% survival of the animals challenged with the mouse-adapted influenza A/Aichi virus. Clinical trials of polyguacyl in human volunteers demonstrated the safety and good tolerance of this drug given both as aerosol and intranasally. PMID- 6272493 TI - [Persistence of Coxsackie A13 virus in the families of children ill with rheumocarditis]. AB - Virological and clinico-instrumental examinations were carried out three times during one year in 13 patients with rheumatic carditis and 34 of their relatives from 13 families. Besides, probands from 5 families had been examined 1-3 years before this study during treatment for rheumocarditis. Among 62 virus strains isolated from feces, blood, and nasopharyngeal washings in the family foci 40 were Coxsackie A13, 12: Coxsackie B1, B2, B5; 10 viruses were not identified. Coxsackie A13 virus was isolated from probands, siblings and parents in 11 out of 13 families. Virus-neutralizing antibodies to it were demonstrated in the great majority of probands and relatives, including those families where no virus had been isolated. Coxsackie A13 virus and antibodies to it were shown to persist throughout the observation period in most families. PMID- 6272494 TI - [Action of hydrolytic enzymes on influenza virus A ribonucleoprotein]. AB - The interaction of protein and RNA in composition of virion ribonucleoprotein (RNP) of influenza A virus was studied by treatment of this structure with hydrolytic enzymes: pancreatic RNase A, nuclease S1 and pronase. The results indicate that RNA in RNP does not shield proteins from the effect of pronase. The possibility of using this fact in the construction of a model RNP structure is discussed. No differences in the effect of RNase A and nuclease S1 on RNP were found which corresponded to the concept of the protective role of RNP protein in the process of RNA hydrolysis. PMID- 6272495 TI - [Enterovirus survival in water and food products]. AB - Inactivation of enteroviruses in water (sewage, river, tap) and some food products (milk, meat, bread) was studied. The process of inactivation was judged by the amount of virus (in PFU) in specimens which were stored under preset experimental conditions and selected during one month at 2--3 day intervals after the virus introduction into the object. The pattern of inactivation was judged by exponents constructed from the experimental results. The time during which half of the initial virus amount was lost has been taken for the unit of the virus inactivation rate in the object. The dynamics of inactivation was found to depend upon the object inoculated with the virus, temperature and the virus strains. Thus poliomyelitis virus survived longer in milk than in meat; its highest inactivation occurred at 37 degrees C (the highest temperature tested); wild poliovirus strains were inactivated slower than the vaccine ones. PMID- 6272496 TI - [Biological properties of the Rous sarcoma virus isolated from transformed hamster cells]. AB - Rous sarcoma virus (RSV) isolated from virogenic transformed hamster cells differed from the original Schmidt-Ruppin strain used to inoculate hamster embryo fibroblasts in the capacity to effectively transform mammalian cells, to multiply in these cells, in the reduced reproductive activity in chicken cells. These altered properties were relatively stable since they were retained in the first virus passages in chickens. PMID- 6272497 TI - [2 cases of malignant mesenchymoma of the breast]. PMID- 6272498 TI - [Patentex-Oval - an intravaginal contraceptive agent]. PMID- 6272499 TI - [The effect of neural mediation on the immediate rise in intraocular pressure in glaucomatous eyes after cyclocryotherapy (author's transl)]. AB - As a rule, there is a transient rise in intraocular pressure following cyclocryotherapy. The preoperative administration of retrobulbar anaesthesia to a group of patients significantly reduced intraocular pressure after cyclocryotherapy in this group as compared with a control group without anaesthesia. These results imply that the temporary rise in intraocular pressure after cyclocryotherapy is mediated by a neural pathway. PMID- 6272500 TI - [Applications of the peroxidase-anti-peroxidase technique in dermatology (author's transl)]. AB - The immunohistological findings are reported in 62 cases of malignant lymphomas, pseudolymphomas and malignant histiocytic disorders of the skin. Paraffin embedded tissue was analyzed with the Peroxidase-Antiperoxidase (PAP) Technique for the presence of intracytoplasmic immunoglobulin (IgM, IgA, IgG, Kappa, Lambda) ana lysozyme. It can be stated that the PAP technique appears to be a valuable aid in interpreting and differentiating selected material of cutaneous lymphoproliferative and histiocytic disorders. The method supplements routine histological and histochemical staining procedures. PMID- 6272501 TI - [Pharmacokinetics of ceftizoxim (author's transl)]. AB - Ceftizoxim is a new cephalosporin derivative, which is also effective against anaerobic rods. The tolerability is excellent. Its pharmacokinetics after parenteral administration of 2 g were investigated in 12 healthy male volunteers. The half-life was 1.40 +/- 0.41 hours, the total clearance 194 l/min. The urinary excretion of the biological effective antibiotic amounted to 66.26 +/- 25.71%. These figures are in the same order of magnitude as those of older cephalosporins. PMID- 6272502 TI - Enzymic hydroxylation of benzo(alpha)pyrene at the 6-position by vitamin K1 hydroperoxide and rat lung hydroperoxidase. AB - 1. The role of vitamin K1 in the hydroxylation of benzo(alpha)pyrene (BP) at the 6-position is correlated with the oxidative formation of vitamin K1-hydroperoxide and its subsequent reaction with BP in the presence of soluble rat lung hydroperoxidase. 2. Vitamin K1-hydroperoxide was synthesized for vitamin K1 and its structure verified by mass spectral analysis and enzymic studies. 3. The major product (the 3,6-BP dione) that results from the non-enzymic oxidation of 6 hydroxy-BP was isolated from the enzymic reaction and characterized by spectroscopic studies. 4. Thiodione (the menadione-glutathione adduct) inhibits both aryl hydrocarbon hydroxylase activity and vitamin K1 hydroperoxide/hydroperoxidase activity. PMID- 6272503 TI - The effect of zeranol implantation on some endocrine glands and gonads in fat tailed lambs. AB - The present work was carried out to study the effect of zeranol on the pars distalis of 39 male fat-tailed lambs about 8 months old. The examined animals were classified into 3 groups. The group I was considered as control, group II and III were implanted subcutaneously in the back side of ear with 3 doses each of 12 mg zeranol at 40 - days intervals. Groups I and II were fed on concentrate mixture containing 70.45% starch and 14.42% digestible protein, while that of the group III containing 70.44% starch value and 7.92% digestible protein. Zeranol implants have been shown to increase body weight of animals group II and III. A significant difference was found in the number of the pars distalis cells as well as the histochemical changes in their properties have been observed in zeranol implanted lambs. The STH-cell showed a significant increase in number and signs of cellular hyperactivity. The ICSH-cells, FSH-cells and TSH-cells were significantly decreased in number and exhibited signs of less activity compared with the propriate cells of the nonimplanted animals. It is concluded that Zeranol improve gain, stimulate STH-cells and inhibit the GTH-cells as well as TSH-cells in the pars distalis of zeranol implanted animals. PMID- 6272504 TI - [The early development of rabbits thymus (author's transl)]. PMID- 6272505 TI - Ascorbate oxidase and its possible involvement in cancer. AB - The electron spin resonance (ESR) spectrum of erythrocytes of a health male volunteer exhibited 2 min after intravenous administration of 1g of ascorbic acid a considerable increase in spin concentration and an new signal at about g-2.005 which we previously had found to correlate to the semidehydroascorbate (SDA) radical and which is not identical with the O2 radical. Moreover, the vitamin C concentration in erythrocytes and plasma was considerably higher than in comparable samples of other volunteers treated identically. In the latter cases, the ESR spectrum of the erythrocytes was not modified at all. These findings suggest that there must be a substance which reacts with ascorbic acid specifically. It can be assumed that the enzyme ascorbate oxidase plays this decisive role in the ascorbic acid metabolism. For this reason, different amounts of ascorbate oxidase have been added to healthy erythrocytes treated in vitro with ascorbic acid and to tissue samples of lung cancer. As expected, the vitamin C effect as expressed by the appearance of the SDA signal and the increase in spin concentrations could be reversed. It is suggested, therefore, that in special types of cancer, such as acute lymphatic leukemia and lung cancer, the concentration of ascorbate oxidase or of an enzyme acting like it is, primarily, diminished, while in other types it might be enlarged. PMID- 6272506 TI - Inhibition cyclic guanosine 3':5'-monophosphate of the soluble DNA polymerase activity, and of partially purified DNA polymerase A (DNA polymerase I) from the yeast Saccharomyces cerevisiae. AB - DNA polymerase activity from extracts of growing yeast cells is inhibited by cGMP. Experiments with partially purified yeast DNA polymerases show, that cGMP inhibits DNA polymerase A (DNA polymerase I from Chang), which is the main component of the soluble DNA polymerase activity in yeast extracts, by competing for the enzyme with the primer-template DNA. Since the enzyme is not only inhibited by 3', 5'-cGMP, but also by 3',5'-cAMP, the 3'--:5'-phosphodiester seems to be crucial for the competition between cGMP and primer. This would be inconsistent with the concept of a 3'-OH primer binding site in the enzyme. The existence of such a site in the yeast DNA polymerase A is indicated from studies with various purine nucleoside monophosphates. When various DNA polymerases are compared, inhibition by cGMP seems to be restricted to those enzymes, which are involved in DNA replication, DNA polymerases with an associated nuclease activity are not inhibited, DNA polymerase B from yeast is even activated by cGMP. Though some relations between the cGMP effect and the presumed function of the enzymes in the living cell are apparent, the biological meaning of the observations in general remains open. PMID- 6272507 TI - [Analgesics and pain therapy]. PMID- 6272508 TI - [Problems and prospects of vitiligo study]. PMID- 6272509 TI - [Plastic changes in responses of chemo- and electro-excitable membranes of a cortical neurons following orthodromal tetanization]. AB - Orthodromic tetanization produces in the turtle visual cortex two types of reversible plastic changes of electrical responses: facilitation and depression. Posttetanic facilitation is attended with reversible enhancement of evoked postsynaptic potentials and APs; the excitability of the electroexcitable membrane increases: the critical level of depolarization of the evoked spike shifts in the negative direction and its threshold potential decreases. Posttetanic depression is accompanied by a reversible decrease of the amplitude of evoked postsynaptic and spike potentials; the excitability of the electroexcitable membrane decreases: the critical level of depolarization of the evoked spike shifts in the positive direction, and its threshold potential increases. Reversible changes of excitability of the spike generating membrane following tetanization testify to the existence of an active mechanism of spike discharge change. PMID- 6272510 TI - [Female functional urinary incontinence]. PMID- 6272511 TI - [Extended medical care for patients with ovarian carcinoma (author's transl)]. AB - Reported in this paper is a scheme of extended medical after-care for patients with ovarian carcinoma at the authors' outpatient department. An account is given of special consultations with routine care as well as individual postoperative treatment, using cytostatics and hormones. A strong demand is made for centralised treatment of patient with ovarian carcinoma which should cover the entire population within reach of the hospital concerned. Such centralisation also should be backup with sufficient personnel, equipment, and laboratory capacity. PMID- 6272512 TI - [Use of ristomycin diffused into the culture medium for separation of meningococci from nasopharyngeal mucus]. AB - The method for enhancing the growth of meningococci after the inoculation of the specimens of nasopharyngeal mucus is proposed. The method is based on the diffusion of ristomycin into agar from ristomycin-impregnated filter paper strips. During the study of 694 specimens of nasopharyngeal mucus the above mentioned method allowed isolation of 108 meningococcal cultures, while in serum agar only 21 cultures were isolated. The effectiveness coefficient of this method was found to be 80.6%, its effectiveness index being 5.1. The method facilitated the isolation of meningococci and in some cases reduced the time of analysis by 1 day. PMID- 6272513 TI - [32P'incorporation into polyphosphoinositides of erythrocytes from essential hypertension]. AB - The level of polyphosphoinositides (PPI) and the incorporation rate of 32P into PPI of erythrocytes from essential hypertensive patients was studied. The PPI were found to have a decreased level and an increased 32P incorporation rate compared with the controls. PMID- 6272514 TI - [Effect of the partial substitution of membrane cholesterol by other sterols on the Na+ + K+ -ATPase activity in microsomal fractions of the cerebral cortex of rats]. PMID- 6272515 TI - Regulation of humoral immunity in rats by pituitary hormones. AB - Hypophysectomized female Fischer 344 and Wistar-Furth rats had severely impaired primary and secondary antibody responses to sheep red blood cells (SRBC). Mercaptoethanol-sensitive (IgM) and mercaptoethanol-resistant (IgG) antibodies were similarly affected. Titers to E. Coli 055:B5 lipopolysaccharide were also significantly decreased in such animals. The antibody response of hypophysectomized rats could be restored by syngeneic pituitary grafts when placed under the kidney capsule or by prolactin treatment. Growth hormone was less effective in this respect than prolactin. Treatment of normal rats with ACTH suppressed their antibody formation to SRBC. These results indicate that the pituitary gland has the potential to regulate humoral immune responses. PMID- 6272516 TI - Presence of gangliosides in the structure of membrane receptor for thyrotrophin. AB - Thyroid gangliosides were isolated from bovine thyroid tissue. Neutral lipids and glycolipids were removed by chromatography on a DEAE-cellulose column and residual acidic phospholipids by alkaline degradation. Thyroid gangliosides were separated by chromatography on a silica acid column into 9 different fractions. Thyroid gangliosides were found to be active in the inhibition of [125I]TSH binding to thyroid plasma membranes and this results from a direct action between thyrotrophin and gangliosides. The most active fraction contained gangliosides with two sialic acids per molecule. Pre-incubation of thyroid plasma membranes for 4 hours at 37 degrees C with gangliosides and removal of the unbound gangliosides increased the [125I]TSH binding to pre-treated membranes by 10-20%. Anti-ganglioside antibodies obtained by immunisation of rabbits inhibited [125I]TSH binding to thyroid plasma membranes by 60 to 80%. The thyroxine (T4) level in blood serum of immunized animals was significantly decreased after the first immunization and T4 become almost undetectable after the booster injection implying that antibodies against thyroid gangliosides produce severe impairment of thyroid cell function. PMID- 6272517 TI - The effect of varying iodine content on the proteolytic activity of rat thyroid lysosomes. AB - The possibility that the iodine supply modulates thyroid lysosomal activity was investigated in rats receiving chronic or acute increasing doses of iodide. The lysosomal activity or the various experimental groups was determined by measuring the activity of beta-glycerophosphatase and cathepsin D both in thyroid homogenates and in semipurified thyroid lysosomal preparations, and the degradation of labelled thyroglobulin by the various lysosomal fractions. In the chronic experiments beta-glycerophosphatase and cathepsin D activities increased with the iodide supply of the animals up to 100 micrograms I/rat and decreased slightly thereafter. In the acute experiments the activity of these enzymes increased up to 1000 micrograms I/rat and decreased above 5000 micrograms I/rat. The proteolytic activity of lysosomal fractions from the various experimental groups towards thyroglobulin decreased slightly with increased iodide supply both in chronic and acute experiments. The results suggest that thyroid lysosomal activity may participate in the autoregulation of thyroid secretion by inducing synthesis of new enzymes and modulating thyroglobulin degradation. PMID- 6272518 TI - The recovery of the hypothalamo-pituitary-adrenal axis after transsphenoidal operation in three patients with Cushing's disease. The effect of prior external pituitary irradiation. PMID- 6272519 TI - The cytoplasmic mucin in Paget cells of extramammary Paget's disease. AB - The cytoplasmic mucin in the Paget cells of extramammary Paget's disease was examined with a battery of histochemical techniques. The staining methods used were alcian blue, azure A and periodic acid-Schiff. In a further attempt to identify various polyanions, staining was carried out with alcian blue containing various concentrations of electrolyte. Methylation, saponification, borohydride reduction, acid hydrolysis, and digestion with diastase, sialidase, chondroitinase ABC, or nucleases were also employed. The results obtained suggest that the cytoplasmic mucin in the Paget cells is sialomucin without side-chain substituent. PMID- 6272520 TI - [A study of occupational vibration-induced disease - correlation between digital angiographic findings in patients mainly with occupational chain saw vibration induced disturbance and sensory disorder (sensation of digital numbness), and somatosensory evoked response (SER) (author's transl)]. PMID- 6272521 TI - [Therapy of villous diseases, particularly that of hydatidiform, and study on their prognoses (author's transl)]. PMID- 6272522 TI - [Production of experimental ovarian tumor in mice by local irradiation of 60Co gamma ray--analysis of the tumor-producing process (author's transl)]. PMID- 6272523 TI - [A study on heterotransplantation of human stomach carcinoma to the nude mouse.(author's transl)]. PMID- 6272524 TI - [Clinicopathological study on the significance of combined pancreatosplenectomy for gastric carcinoma -Especially, gross indication on the base of oncological features (author's transl)]. PMID- 6272525 TI - LH(hCG) receptor in benign and malignant tumors of human ovary. AB - LH(hCG) receptors were identified with [125I]hCG in 38 benign and 26 malignant ovarian tumors of the human ovary. Eighteen per cent of all the benign and 27% of all the malignant tumors were LH(hCG) receptor-positive. Four of 12 benign serous tumors and 3 of 17 benign mucinous tumors displayed definitive binding of [125I]hCG. Two Brenner tumors failed to bind [125I]hCG. Six of 21 malignant epithelial tumors displayed definitive binding of [125I]hCG. Only one out of 4 malignant granulosa cell tumors bound [125I]hCG, while the other sex cord stromal tumors as well as one dysgerminoma failed to bind [125I]hCG. LH(hCG) receptor content in the benign and malignant receptor-positive tumors was low compared with the normal ovarian tissue. The presence of LH(hCG) receptors in certain benign and malignant ovarian tumors may be a sign of the gonadotropic control of these tumors. The possible applications of these findings for the diagnosis and treatment of human ovarian tumors is discussed. PMID- 6272526 TI - Enzyme immunoassay for human chorionic gonadotropin and its clinical application. AB - The assay for hCG is important in gynecological and obstetrical fields and has been performed by RIA and radioreceptor assay (RRA). But these procedures use radioisotope which limits popular use. Recently enzyme immunoassay (EIA) has been developed in which procedure the enzyme has been used for labelling in place of radioisotope. The labelling enzyme is stable and has no adverse effect on human beings and its activity is easily measured. In this study, EIA for hCG has been developed. beta-D-Galactosidase (beta-Gal) is preferred because an infinitesimal amount of beta-Gal is easily measured, and MCAE of conjugating agent is used to prevent self-coupling. As solid phase sheep red blood cells were used at first, but now silicone rods are being used because of easy handling. To remove serum interference, sequential competitive method is carried out and 0.5 mIU/ml to 200 mIU/ml of hCG is measurable in 12 h. Coefficients of variation are satisfied. This newly developed assay has been used clinically in diagnosis of brain metastasis of trophoblastic disease, early diagnosis of pregnancy and prognosis of threatened abortion and is now being tried for follow-up of trophoblastic disease. PMID- 6272527 TI - [An ultrastructural and cytochemical study of human corpora lutea (author's transl)]. AB - Human luteal cells have characteristic organelles, such as well developed smooth endoplasmic reticulum (sER), large mitochondria with tubular cristae, lysosome like granules and lipid droplets. To investigate possible roles of these characteristic organelles, 21 human active corpora lutea, obtained from women aged 32-46, were studied cytochemically on 3 beta-hydroxysteroid dehydrogenase (3 beta-HSD), cytochrome c oxidase (CY-O), NADPH diaphorase (NP-DP) and acid phosphatase (ACP) activities, using the methods by Benkoel et al., Ogawa et al., Ago et Ogawa and Barka et Anderson. Subcellular localization of 3 beta-sterol were also examined using the 3 beta-sterol digitonin complex method by Mizuhira et al., 3 beta-HSD and NP-DP activities were demonstrated in the intercrystal and outer space of mitochondria and sER. CY-O activities were demonstrated in mitochondria. ACP activities were demonstrated in lysosomes and Golgi apparatus. Some lipid droplets contained the crystals of 3 beta-sterol digitonin complex. These data suggest that human luteal mitochondria can synthetize progesterone from cholesterol, and that mitochondrial respiratory chain activities in human luteal cells is highly elevated. Human luteal cells probably require much oxygen. Elevated ACP activities indicate that lysosomes play a role in the steroidogenesis. Some lipid droplets may contain cholesterol and other 3 beta sterols. PMID- 6272528 TI - [Morphological and endocrinological study of ovarian arrhenoblastoma (author's transl)]. AB - Morphological and endocrinological studies were performed on a 19-year-old case of an arrhenoblastoma with marked virilization. The tumor was an intermediate type of Meyer's classification. Histochemically, 3 beta-HSD and G-6-PDH activities were demonstrated in Leydig cells. These cells also had ultrastructures typical of steroid-producing cells. Basal blood cells of pregnenolone, dehydroepiandrosterone, testosterone (T), androstenedione (A), progesterone, estradiol, estrone, LH, and FSH were determined pre- and postoperatively. T and A showed a very high level preoperatively, and were markedly decreased immediately after removal of the tumor. Stimulation of tumor cells by HMG-HCG did not show any significant changes in their main products of T and A. These findings suggest that Leydig cells of the present tumor produced mainly A and T independently on gonadotropins, and these hormones had virilized the patient. PMID- 6272529 TI - Treatment with oral estrone sulphate in the female climacteric. III. Effects on bone density and on certain biochemical parameters. AB - Thirty-eight women were treated with estrone sulphate over a period of 30 months for climacteric problems. A random control group of 29 women were given methyl scopolamine. Bone mineral assays were performed by means of dual photon absorptiometry before the start of treatment and after 6, 12, 18, and 30 months. There was a difference in the mineral loss of trabecular bone between the two groups after 30 months of treatment, that the control group being significantly higher (p less than 0.001). In the estrogen-treated group there was a decrease in serum phosphorus (p less than 0.01), alkaline phosphatase activity (p less than 0.001), and albumin (p less than 0.001). In the control group, serum albumin showed the same decrease, while the other factors either showed no differences or even increased. The urinary excretion of calcium was not significantly reduced in the estrogen group, whereas there was an increase in the control group (p less than 0.01). There was an increase in hematocrit in both groups. Positive correlation was found between parity and loss of trabecular bone mineral (p less than 0.01).U PMID- 6272530 TI - A case of hepatoma in pregnancy associated with earlier oral contraception. PMID- 6272531 TI - The orbit and eye: experiments on volume in young and adult rabbits. PMID- 6272532 TI - Ultrasound in the diagnosis of maxillary and frontal sinusitis. PMID- 6272533 TI - Inert gas exchange in the middle ear. AB - A novel mathematical description of inert gas exchange in the middle ear has been formulated and related to experimental measurements of the middle ear exchange of helium, neon, and nitrous oxide in the cat. This study indicates that the diffusion of gases across the tympanic membrane is not significantly affected by its blood supply and that the diffusion of gases across the mucosal layer of the middle ear cavity in perfusion limited. The diffusion of helium and neon is consistent with an aqueous model of the tympanic membrane, while that of nitrous oxide is one-tenth of the expected value for an aqueous membrane. PMID- 6272534 TI - Critical review of the concept of cupula function. PMID- 6272535 TI - Hearing in classical musicians. PMID- 6272536 TI - Comparison of cardiovascular responses in noise-resistant and noise-sensitive workers. PMID- 6272537 TI - Mechanisms underlying modulations of thermal nystagmic responses in parabolic flight. AB - In six subjects nystagmography was used to compare the responses to cold calorization of one horizontal semicircular canal under ground-based and parabolic flight conditions. On the ground the expected individual differences in primary nystagmic responses were observed; only one subject manifested a brief weak secondary nystagmus. Aloft, the irrigation was carried out in straight-and level flight prior to a pushover (half-parabola) that initiated a series of four to nine typical parabolas in a modified KC-135 aircraft. Thereafter, the free fall phases of the parabolas furnished a zero baseline for measuring weight differences in endolymph due to changes in gravitoinertial force. In all subjects a secondary nystagmus was generated in addition to the primary nystagmus during the course of the parabolic maneuvers. The slow phase of the secondary nystagmus rarely exceeded 10 mm per sec. Evidence is presented that secondary nystagmus arises as a direction-specific adaptation effect countering the primary nystagmic response. Three stages were recognizable: first, when primary and secondary nystagmus alternated in step with the high and low force phases of the parabolas; second, after disappearance of primary nystagmus when secondary nystagmus appeared alone and was modulated by the changes in force; and third, when the secondary nystagmus present became independent of the highest gravitoinertial forces generated. Great individual differences were observed, suggesting that with large departures of the cupula from its functional rest position there are large individual variations in rate of restoration. PMID- 6272538 TI - [Corticoid myopathy in children]. PMID- 6272539 TI - Efficiency of tetanus toxoid booster in leukaemic children. PMID- 6272540 TI - Serological follow-up of children with infectious mononucleosis caused by Epstein Barr virus. AB - Ninety children, most under 6 years of age, 73 of whom were suffering from a disease suspect of infectious mononucleosis, were examined serologically for EBV infection. Acute EBV infection was demonstrated in 40 cases. These children were followed up until the end of the second year by examining serum samples taken every month of every third month for anti-VCA-IgM, anti-VCA-IgG, anti-VCA-IgA anti-EBNA, and for the specific antibodies to the components D and R of the early antigen. Heterophil antibodies were examined by agglutination of sheep and horse erythrocytes. The temporal course of the antibody response was similar to those reported in adults while anti-EA-D and heterophil antibodies were demonstrated less frequently. PMID- 6272541 TI - Histopathological identification of hepatic intramitochondrial crystalloid inclusions (IMCI). AB - A 45-year-old female with systemic lupus erythematosus underwent liver biopsies twice during the period of cefazolin administration and a complication of cefazolin-induced toxic hepatitis was strongly suggested. Histologically, a small number of spindle-shaped, esoinophilic crystalloid bodies were faintly noticed in the cytoplasm of hepatocytes in hematoxylin and eosin-stained sections, while they showed strong positivity for strains inherent to mitochondria as well as specific to phospholipid and sharply showed a spindle or needle shaped configuration. Moreover, they greatly increased in number in these sections. They were distributed throughout the the parenchyma but were prominent in the degenerated hepatocytes with slight fatty metamorphosis. Electron microscopically, they proved to correspond to the intramitochondrial crystalloid inclusions (IMCI). Despite many ultrastructural descriptions of the IMCI, precise histopathological examination of the IMCI referring to the staining property on the deparaffinated sections has not yet been described since the first report of the IMCI by Napolitano (1958). PMID- 6272542 TI - Hepatocellular carcinoma combined with hepatic sarcoma. AB - An autopsy case of hepatocellular carcinoma combined with hepatic sarcoma was presented. There were three main tumor nodes in the liver. They showed necrosis due to therapeutic embolization of the hepatic artery. Histologically, one of the three was a hepatocellular carcinoma, and the other two were of spindle cell sarcomas. Extrahepatic metastases all showed characteristics of sarcoma. In the present case report, some problems concerning hepatic sarcoma and sarcomatous representation of hepatocellular carcinoma were proposed in reviewing the literatures. PMID- 6272543 TI - Type 1 glycogenosis with contracted kidneys and liver cell adenoma. AB - A 22-year-old man with type 1 glycogenosis died of renal and respiratory failure. Postmortem examination revealed deposition of glycogen in liver cells, a liver cell adenoma, bilateral contracted kidneys with scent glycogen deposition, and pulmonary edema. The development of liver cell adenoma was thought to be related to underlying metabolic disorder and contracted kidneys were considered to be the sequel of massive glycogen deposition in the renal tubular epithelium. PMID- 6272544 TI - Hormones, receptors and cyclic nucleotides. Papers presented at the 6th Scandinavian Meeting on Cyclic Nucleotide Research. Uto, May 24-26, 1981. Abstracts. PMID- 6272545 TI - Effect of pituitary intermediate lobe extract on steroid production by the isolated zona glomerulosa and fasciculata cells. AB - The effect of intermediate lobe extract (ILE) on aldosterone and corticosterone production of the zona glomerulosa cells and on corticosterone production of the zona fasciculata cells was investigated. The slope of the dose-response curve of ILE dilution was steeper than that of alpha h 1-39 ACTH measured on zona glomerulosa steroid production. The ED50 of both ILE and ACTH was lower when measured on zona glomerulosa than on zona fasciculata steroid production. It is supposed that a hormone (or some other substance) in ILE alters the sensitivity to ACTH of the zona glomerulosa cells. PMID- 6272546 TI - The increase of cerebellar cAMP level after decapitation: the effect of propranolol. AB - The increase of cAMP level of the rat cerebellum induced by decapitation was studied. Administration of 5 mg/100 g propranolol 1 hour before decapitation completely prevented this increase. Neither the depletion of catecholamine pools, inhibition of their synthesis, nor barbiturate treatment influenced the increase of cAMP level evoked by decapitation. It has been concluded that noradrenergic neurotransmission is involved in the cerebellar cAMP level increase after decapitation. PMID- 6272547 TI - The level of blood kinins, their creation and inactivation after one exposure to intense ultraviolet radiation in rabbits. AB - Experimental rabbits were exposed to ultraviolet radiation once for 45 minutes, and blood samples were obtained from the carotid artery in these animals 45 min and 3, 6 and 24 hours after the end of this exposure. In the group of control rabbits blood samples were obtained in the same way without previous exposure to radiation. The hairs on the back were cut closely at the skin and this skin area was exposed to ultraviolet rays from a Hanau Q 400 burner at 405--289 nm wavelengths and at an intensity of 134 000 erg/sec/cm2, using an UG 2 T Schott filtre and an absorber of long-wave radiation. Blood samples were taken under thiopental anaesthesia. In the samples the level of free kinins was determined in the blood, and the level of kininogen and the activity of kallikreins and kininases were determined in the plasma. In the irradiated animals a rise of the kinin level was observed, with a fall in the kininogen level most pronounced after 3 hours, while the activity of kallikreins was raised and that of kininases was decreased particularly after 6 hours. PMID- 6272548 TI - Effect of long-term exposure of rabbits to ultraviolet radiation on the level, creation and inactivation of kinins in blood. AB - Experimental rabbits were exposed to ultraviolet radiation during 6 weeks once daily for 10 minutes (from a high-pressure. Hanau Q 400 mercury lamp with a Schott UG 2 T filtre and absorber of long-wave radiation using ultraviolet rays of 405--289 nm wavelength and 134 000 erg/sec/cm2 power, directed onto skin with cropped hair of the back). Under general anaesthesia with pentothal blood samples were obtained from the carotid artery in 4 groups of 8 rabbits in each group. The blood samples were taken from non-exposed control rabbits and from the experimental groups after 2, 4 and 6 weeks of exposure. In the samples the levels of free kinins in the blood, and kininogen, and the activity of kallikreins and kininases in the plasma were determined. In the irradiated animals a progressive rise of free kinins most pronounced after 6 weeks was observed, and other findings included: a fall of kininogen level particularly steep after 2 weeks, very small rise in the activity of kallikreins, and progressing reduction of the activity of kininase, particularly steep after 2 weeks. PMID- 6272549 TI - Abolition by preventive action of calcitonin of vitamin D3-stimulated gluconeogenesis in rat kidney. AB - The effects of vitamin D3, 1-alpha-OH D3 and calcitonin treatment on the rate of glucose synthesis in rat renal cortex slices were studied. The rate of glucose synthesis was significantly increased in animals treated with vitamin D3 and 1 alpha-OH D3 as compared to the control. The increase in the rat of glucose synthesis observed after vitamin D3 and 1-alpha-OH D3 treatment was abolished after administration of calcitonin in doses normalizing the serum calcium level. The obtained data point to a correlation between the serum calcium level observed in vivo and the ability to synthesize glucose measured in vitro in renal cortex slices. PMID- 6272551 TI - Radiologic and prognostic aspects of Wilms' tumour. AB - Retrospectively the urographic findings, history, clinical signs and surgical/pathologic features of 46 Wilms' tumours were analysed. Imparied or no function of the kidney was associated with a high incidence of defects of the renal pelvis and invasion of the renal vein. A high rate of non-radically operated or inoperable tumours and high mortality was present in these cases. PMID- 6272550 TI - Reversal of alpha 1-receptor mediated relaxation in intestinal smooth muscle. AB - The alpha 1-receptor agonist phenylephrine relaxed longitudinal rabbit jejunal muscle contracted in vitro by low concentrations of barium ions (1 mM). When the Ba2+ concentration was increased to 10-15 mM the response to phenylephrine was a contraction, and at Ba2+ concentrations in between the high and low range this response was biphasic--a relaxation followed by a contractile phase. The alpha 2 receptor agonist clonidine did not affect the tone of the Ba2+ contracted preparation. When the muscle preparation was contracted by Sr2+ (1-20 mM) in the presence of Ca2+ (2.5 mM), phenylephrine relaxed it, and no contractile response to phenylephrine was observed. In the absence of extracellular Ca2+, 5 mM Ba2+ caused a contraction. Under these conditions phenylephrine had no effect on the tissue tone. When Ca2+ was added in a low concentration (0.2-2 mM), phenylephrine elicited a gradually increasing contractile response. At 5 mM Ca2+ the contractile response was replaced by the normal relaxation. The contractile response to phenylephrine in the presence of 5 mM Ba2+ and 2.5 mM Ca2+ was partially blocked by low concentrations of verapamil. In higher concentrations verapamil abolished the tissue tonus completely. The contractile response to phenylephrine in the presence of 5 mM Ba2+ and 2.5 mM Ca2+ could be reverted to the normal relaxation by the addition of 20 mM Mg2+. Increasing the K+ concentration from the normal 5.9 to 62.9 mM blocked the phenylephrine-induced relaxation. No contractile response to phenylephrine occurred. It is concluded that Ba2+ could reverse the response of alpha 1 receptor stimulation in rabbit jejunum from a relaxation to a contraction and that this contractile response was dependent on the presence of Ca2+. PMID- 6272552 TI - Nucleoside analogues as antiviral agents. PMID- 6272553 TI - Differential inhibition of herpes simplex viruses, type 1 (HSV-1) an type 2 (HSV 2), by (E)-5-(2-X-vinyl)-2'-deoxyuridines. PMID- 6272554 TI - Morphometric aspects and dynamics of lipofuscin granules in "torpedo m." neurons. PMID- 6272555 TI - Corticosteroid treatment in patient with "empty sella syndrome". PMID- 6272556 TI - Cardiac evaluation using nuclear medicine procedures. PMID- 6272557 TI - Local cerebral blood flow and tissue solubility measured by stable, xenon enhanced computerized tomography. PMID- 6272558 TI - Dynamic computed tomography study of the brain. AB - The dynamic computed tomography study of the brain consists of the rapid intravenous injection of 49 ml of radiographic contrast material in 7 sec followed by serial 5-sec CT scans with interscan times of only 1 sec. The data from these scans can be reprocessed to create 12 segmented images in 35 sec. When small samples of four or six pixels of cortex are examined by the cursor, sharp rises of 20 to 25 CT units (500 scale) are seen on the time-density curves. Samples of white matter are usually no more than two CT units. When larger cursor samples of 220 to 255 pixels including cortex and white matter are examined, the time-density curves represent a combination of these two patterns. Comparison between symmetrical areas in the two hemispheres generally shows parallel curves in controls. The studies provide high-resolution cerebral perfusion images. In ischemia secondary to ipsilateral carotid stenosis, there is depression of the up slope and a depressed late peak. Infarctions show a flat perfusion curve. Several patterns in brain tumors have been illustrated. The study, simple to perform, adds significant perfusion information to the previously static CT examination of the brain. PMID- 6272559 TI - The use of substrates and energy in the treatment of shock. AB - Various metabolic, cellular, and subcellular alterations in cell function and morphology occur during shock or low-flow conditions. In attempting to find treatment programs that would be beneficial following shock, various substrates have been used. Infusion of hypertonic glucose during shock has been shown to improve survival; however, it is unlikely that the effect of glucose is by provision of energy until the circulation is restored. Infusion of glucose- insulin--potassium during shock has also been reported to be beneficial in certain clinical situations. Controversies exist concerning the efficacy of infusions of cyclic AMP, nicotinamide, and Krebs cycle intermediates during shock. Pretreatment of kidneys with inosine or raising glycogen stores of the myocardium have been shown to have protective effects of kidneys and myocardium during ischemia and these procedures may be suitable for organ preservation. Pretreatment with allopurinol has been shown to be beneficial in shock; however, it is unlikely that allopurinol by itself if given following shock would have any salutary effects. Treatment with ATP-MgCl2 has been shown to be beneficial following hemorrhagic shock, sepsis, endotoxin shock, burns, postischemic hepatic failure, and postischemic renal failure. Thus, provision of energy directly in the form of ATP during adverse circulatory conditions appears to be the most advantageous and direct method for the treatment of shock. PMID- 6272560 TI - Luteal blood flow and receptors for LH during PGF2 alpha-induced luteolysis: production of PGE2 and PGF2 alpha during early pregnancy. PMID- 6272561 TI - Corpus luteum prostaglandin receptors and luteolysis. PMID- 6272562 TI - Effects of calcium and sodium concentrations on atrioventricular conduction: experimental study in rabbit hearts with clinical implications on heart block and slow calcium channel blocking agent usage. AB - Effects of electrolyte concentrations on atrioventricular (AV) conduction were studied in isolated, perfused rabbit hearts by recording the His bundle electrogram. Initial calcium (Ca++), sodium (Na+), and potassium (K+) concentrations were 2.4, 144.8, and 4.5 mM, respectively. Lowering of Ca++ to 0.8 mM slightly prolonged the AH interval, whereas elevation of Ca++ to 4.8 or 7.2 mM more markedly prolonged this interval, often causing intranodal block. High Ca++ induced depression of intranodal conduction was antagonized by high K+ (7.5 mM). Verapamil (0.5 to 1.0 mg/L) produced second-degree intranodal block. High Na+ (172 mM) restored 1:1 conduction, whereas high Ca++ did not. These results suggest that: (1) an optimal Ca++ concentration for intranodal conduction exists; (2) high K+ counteracts high Ca++-induced intranodal block; (3) verapamil effect on AV node is antagonized by high Na+; and (4) slow Na+ current may play a role in AV nodal action potentials. PMID- 6272563 TI - Effects of diltiazem on conduction of premature impulses during acute myocardial ischemia and reperfusion. AB - The effect of diltiazem on conduction of cardiac impulses was studied using premature impulses. Conduction times were measured in the epicardium and endocardium in both anterograde and retrograde directions during ischemia and reperfusion of the left ventricular anterior wall. In 16 dogs (8 control and 8 treated with diltiazem), the left anterior descending artery was occluded initially below the second diagonal branch and 30 minutes later below the first diagonal branch. Infusion of diltiazem (0.02 mg/kg per min) was begun at the time of the first ligation in the treated dogs. Conduction delay in normal, ischemic and reperfused myocardium and at the border of ischemia or reperfusion was compared in the two groups. In addition, treated and nontreated ischemic zones were analyzed in the diltiazem-treated group. Results show significantly less ischemia-induced conduction delay in the diltiazem-treated group in both ischemic myocardium and at the border of ischemia. This beneficial effect was seen in the ischemic segment in the treated dogs whether or not the medication was given before or after coronary ligation. Further, the effect of the drug on epicardial and endocardial conduction did not differ significantly. These data suggest that diltiazem may have potential value in the treatment of ventricular reentrant arrhythmias associated with acute ischemia over and above the well recognized hemodynamic and metabolic effects of calcium channel blocking agents. PMID- 6272564 TI - Two dimensional echocardiography in evaluation of right atrial masses: five cases in pediatric patients. AB - Five unusual cases of a right atrial mass in children are described to illustrate the very valuable contribution that two dimensional echocardiographic examinations can bring to both the initial diagnosis and the subsequent management of patients with these findings. One patient had a large benign hemangioendothelioma of the right atrium. Two infants had extension of a Wilms' tumor from the kidney by way of the inferior vena cava to the right atrium. A fourth patient, an 8 year old girl, had no cardiac disease, and manifested Staphylococcus aureus endocarditis of the tricuspid valve with a large pedunculated mass and subsequent pulmonary embolus. A fifth patient, a premature infant with a central hyperalimentation catheter in the right atrium, had a large thrombus on the catheter that was successfully eradicated with urokinase-induced thrombolysis. Two dimensional echocardiography provides real time imaging of the entire right atrium, interatrial septum, inferior and superior venae cavae and tricuspid orifice and hence is valuable in the diagnosis and management of these clinical problems. PMID- 6272565 TI - Effect of fiber in sorghum on nitrogen digestibility. AB - Digestibility measurements were carried out on a population of 12 Cameroonians whose habitual diet, deficient in animal products, is based on consumption of sorghum meal which supplies between 2.4 and 4.2 g of crude fiber per 100 g of dry matter. Over an 11-day period, the 12 subjects received successive diets of 3.3, 4.8, and 5.4 g of crude fiber per 100 g of dry matter. The increase in fiber intake resulted in a significant rise in quantity of fecal matter excreted, including nitrogen and formic insoluble substances. On the other hand, the highest urinary nitrogen losses were obtained from the diet least rich in fiber. The apparent digestibility of nitrogen dropped from 65.4 to 60.5% and then further to 56.9%. The apparent digestibility of nitrogen of the diet least rich in fiber differed significantly from those of the other two diets. The subjects under study did not benefit from an adaptive physiology which would enable them to reduce digestive nitrogen losses. Intestinal disappearance of crude fiber augmented significantly and then stabilized when switching from the diet least rich in fiber to the two others, i.e., from 15.1 to 19.9 g/day and then 19.8 g/day. This population was distinguished by its ability to breakdown large quantities of fiber and to reduce urinary nitrogen loss when fecal nitrogen output rises. Apparent nitrogen balances remained positive. PMID- 6272566 TI - The effect of coarse and fine Canadian Red Spring Wheat and French Soft Wheat bran on colonic motility in patients with diverticular disease. AB - Bran from a Canadian Red Spring Wheat, of both a coarse and fine type, was compared with that from a French Soft Wheat, also with coarse and fine characteristics. The coarse type whether Canadian or French had the more significant effect on the stool weight, speeded the intestinal transit as measured by Hinton markers, and reduced intraluminal pressure in the colon more than did the fine types from the same sources in patients with diverticular disease. The texture of a bran may be important in relationship to its clinical efficacy. PMID- 6272567 TI - Nutrient intake and health status of vegans. Chemical analyses of diets using the duplicate portion sampling technique. AB - A strict vegetarian diet [vegan diet (VD)] was investigated. Six middle-aged vegans (three men and three women) collected copies of 24-h diets using the duplicate portion sampling technique. By chemical analyses, the nutrient composition was determined in detail and compared with corresponding figures of a normal mixed Swedish diet. In the VD 30% of the energy originated from fat compared with 40% in normal Swedish mixed diet (MD). Linoleic acid was the dominant fatty acid (60% of total fat in VD versus 8% in MD). The VD contained 24 g protein/1000 kcal compared to 30 g/1000 kcal in MD, but the intake of essential amino acids by the vegans exceeded the recommendations. Dietary fiber was about 5 times higher in the vegan diet (29 versus 6 g/1000 kcal) and sucrose similar to MD (18 versus 21 g/1000 kcal). Among the inorganic nutrients the concentration of calcium (351 versus 391 mg/1000 kcal) and sodium (53 versus 49 mmol/1000 kcal) were similar in both types of diets but the amount of potassium (56 versus 30 mmol/1000 kcal, magnesium (300 versus 110 mg/1000 kcal), iron (9 versus 6.5 mg/1000 kcal), zinc (6.5 versus 4.7 mg/1000 kcal), and copper (2 versus 0.7 mg/1000 kcal) were nearly doubled. Iodine (39 versus 156 micrograms/1000 kcal and selenium (5 versus 17 micrograms/1000 kcal) were much lower in the VD, selenium even being undetectable in several 24-h diets. The VD was rich in folic acid (301 versus 90 micrograms/1000 kcal in MD) but the intake of vitamin B12 was only 0.3 to 0.4 microgram/day (MD: 3 to 4 micrograms/day). No clinical signs of nutritional deficiency were observed in the vegans. Serum protein levels of the vegans as well as their serum lipoproteins were near the lower range of the reference group. In addition, none of the vegans was overweight and their blood pressures were low for their age. PMID- 6272568 TI - New findings concerning the pathogenesis of acute carbon monoxide (CO) poisoning. AB - The pathomechanism of carbon monoxide (CO) poisoning was studied by means of cardiopathological and neuropathological investigations in experimental CO intoxication. It has been shown that besides CO-hemoglobin association, the binding of CO to cytochromes is a significant factor. The latter is thought to be responsible for the cytotoxic phenomena. Combined ultrastructural and cytochemical studies have enabled differentiation between toxic, hypoxic, and mixed alternations. PMID- 6272569 TI - Clinical aspects of glucagon-producing islet cell tumors. AB - Review of the 55 reported cases of glucagon-producing tumors reveals that a distinctive clinical syndrome consisting of diabetes, a peculiar dermatitis termed necrolytic migratory erythema, weight loss and an increased tendency for thrombosis is associated with these neoplasms. Normochromic normocytic anemia, hypocholesterolemia, hypoproteinemia and generalized hypoaminoacidemia are frequent laboratory findings. Definitive diagnosis of a glucagonoma requires elevation of the fasting serum glucagon level. Selective arteriography of the pancreas has been the best method for localizing these neoplasms preoperatively, but the noninvasive technics of ultrasound and CAT scanning can also be helpful. When the tumor is benign, complete surgical excision can completely reverse all the clinical manifestations of the glucagonoma syndrome and result in lasting cure. Since, however, approximately three-fourths of these tumors are malignant, palliative therapy is frequently required. Cytoreductive surgery can decrease the amount of hormone-producing tissue and can improve or even temporarily reverse the clinical symptomatology. For disseminated disease, chemotherapy is necessary. The best results have been obtained with DTIC although streptozotocin has also been used. PMID- 6272570 TI - A mixed epidemic associated with echovirus types 6 and 11: virologic, clinical and epidemiologic studies. AB - During 1979, an outbreak of mixed enterovirus infections occurred in Kansas City and adjacent communities. Sixty-six enteroviruses and 7 adenoviruses were recovered from 73 persons in a survey hospital population. Twenty-eight persons yielded echovirus type 11, 22, echovirus type 6, and 16 either echovirus type 9 or Coxsackieviruses group B, types 2 and 3. This study describes some of the clinical, virologic and epidemiologic characteristics of the outbreak. A major finding relates to the recovery of enteroviruses from cerebrospinal fluids with essentially normal leukocyte counts. PMID- 6272571 TI - Norwalk-related viral gastroenteritis due to contaminated drinking water. AB - An explosive outbreak of gastrointestinal illness clinically compatible with infection by an agent serologically related to Norwalk virus agent occurred in an elementary school in May 1978. Seroconversion by radioimmunoassay to the Norwalk antigen was noted in two of three ill persons, but no viral particles were identified in stool. Illness developed in 72% of students and teachers at the school, and 32% of household contacts of these ill persons. Of household contacts of persons exposed at school but not clinically ill, 11% developed illness. This value, however, was not statistically different from the level of illness observed concurrently in household contacts of students at an unaffected school nearby. Epidemiologic investigation implicated water as the mode of transmission. Average consumption of one or more glasses per day was strongly associated with illness (p less than 0.00000001). Among soccer team members with limited school contact, water consumption at the school was associated with a 14-fold greater risk of illness (p less than 0.000001). Drinking water was most likely contaminated by back-siphonage through a cross-connection between the school's well and septic tank. This contamination occurred approximately 24 to 36 hours before the outbreak developed. PMID- 6272572 TI - The Alexander D. Langmuir Lecture. The pathogenesis of dengue. Molecular epidemiology in infectious disease. PMID- 6272573 TI - The aggregation of human platelets by ascitic fluid: a possible mechanism for disseminated intravascular coagulation complicating LeVeen shunts. AB - To identifiy causative factors responsible for the disseminated intravascular coagulation complicating peritoneovenous (LeVeen) shunts, the ascitic fluid from 12 patients with alcoholic liver disease or peritoneal malignancy was examined for its effects on human platelets. In all patients, concentrated ascitic fluid caused irreversible platelet aggregation. Properties of the aggregating factor suggested that it is collagen, and subsequently, the presence of collagen in ascitic fluid was confirmed. This finding, together with the known effects of collagen on platelets and contact clotting factors, would be sufficient to explain the development of disseminated intravascular coagulation following this procedure. Aspirin by inhibiting collagen-induced aggregation may have a therapeutic role in the management of this problem. PMID- 6272574 TI - Rapidly progressive silicon nephropathy. AB - Rapidly progressive renal failure developed in four patients with silica exposure. Three presented with manifestations of a connective tissue disorders. All had abnormal proteinuria, hypoalbuminemia and active urinary sediments. Histologically, a distinct constellation of findings was present, consisting of glomerular hypercellularity and sclerosis, crescents, interstitial cellular infiltrates and tubular necrosis with red cell casts as seen on light microscopy. On electron microscopy there was foot process obliteration, characteristic cytoplasmic dense lysosomes, microtubules and dense deposits. Despite vigorous treatment, two patients died of the systemic illness and one is on hemodialysis. The fourth is improved after pulse methylprednisolone therapy. We propose that silica induced this multisystem disease through activation of the immune system and a direct tissue toxic effect. PMID- 6272575 TI - Clinical and laboratory findings and results of therapy in 58 patients with adrenocortical tumors admitted to a single medical center (1951 to 1978). AB - We reviewed 150 findings in 58 patients (14 males and 44 females) with adrenocortical tumors (26 with adenoma and 32 with carcinoma) admitted to Vanderbilt Hospital during 28 years. In general, our findings agree with those reported by others in multi-institutional series or literature reviews. Adenomas took longer to diagnose than carcinomas. Adenomas usually caused Cushing's syndrome, but two caused virilization and three caused no endocrine syndrome. There was no difference in time required for diagnosis of carcinoma in men or women or in those with Cushing's syndrome, virilization or no endocrine syndrome. Urinary 17-hydroxycorticoid (17-OHCS) levels were similar in those with adenoma and those with carcinoma, but 17-ketosteroid (17-KS) levels were usually less than 20 mg per day in patients with adenoma and greater than 20 mg per day in patients with carcinoma. Adenomas were uniformly independent of endogenous ACTH stimulation, but frequently responded to exogenous ACTH. As with adenomas, no carcinoma demonstrated normal suppression with dexamethasone or normal response to metyrapone, but only one responded to exogenous ACTH. Some patients had no clinical Cushing's syndrome despite high levels of plasma cortisol and urine 17 OHCS. "Nonfunctional" tumors probably merely secreted insufficient steroids to cause signs and symptoms. Patients with adenoma were uniformly cured by surgical tumor resection. Occasional patients with carcinoma enjoyed long survival despite incomplete resection of their tumors, but most patients died of recurrent of metastatic carcinoma within seven years, often within a year of two. Small tumor size and benign histologic features were insufficient to predict benign clinical behavior. The adrenocorticolytic drug, o,p'DDD, offered objective remission for only an occasional patient. PMID- 6272576 TI - Symphalangism with multiple anomalies of the hands and feet: a new genetic trait. AB - We report a new autosomal dominant condition involving hands and feet of an Arabic father and 5 of his 11 children. This trait is characterized by symphalangism, syndactyly, brachydactyly type D, clinodactyly, and hypoplasia of the thenar and hypothenar eminences. Affected persons had symphalangism and syndactyly plus some or all or part of the other anomalies. Symphalangism, the main defect in this syndrome, showed variable expressivity. A distinct dermatoglyphic pattern was observed in all affected relatives. Linkage studies were done; however, no linkage was demonstrated. PMID- 6272577 TI - Ocular infectious mononucleosis manifested as Parinaud's oculoglandular syndrome. PMID- 6272578 TI - Human vestibular epithelia: prospects for future study. AB - The lateral ampullae of thirteen patients, removed during translabyrinthine surgery for various disorders, have been examined with light and electron microscopy. Correlation of results of vestibular function tests with changes of histology and ultrastructure have been attempted. Light and electron microscopy revealed the same features that other investigators have reported as pathologic changes in the sensory epithelium and associated subepithelial structures. The occurrence of these features did not correlate with the vestibular function tests, indicating that these features, in our tissue samples, were handling and preparation artifacts. Two observations made in a specimen from a patient with Meniere's disease were difficult to ascribe to handling or preparation artifact. One was the presence of densely stained granular inclusions in the basal portion of the sensory epithelium. The location and contents of these inclusions as seen in the electron microscopy indicated they were hypertrophied degenerating nerve terminals. Similar inclusions were also seen in the axoplasm of myelinated nerve fibers in the sensory subepithelial connective tissue. The other observation believed to be relevant to the symptoms of Meniere's disease, was the presence of widened tight junctions between adjoining sensory and supporting epithelial cells. The attempt at correlation of histologic and ultrastructural changes of ampullae with vestibular function has made possible some differentiation between pathology and artifact. The significance of the ultrastructural observations is discussed. PMID- 6272579 TI - In vitro models of viral-induced congenital deafness. AB - Cytomegalovirus (CMV) infects 1 to 2 percent of liveborn infants in the United States and causes varying degrees of perceptive hearing loss. There are eight reported pathologic studies of temporal bones in CMV-infected neonates. Viral replication occurs in nonsensory endolabyrinthine epithelium, but viral antigen is also found in the organ of Corti and spiral ganglion neurons, and CMV has been cultured from perilymph. Further clinicopathologic correlation is frustrated, since the inner ear cannot be biopsied during life, and the number of temporal bones available for study is limited, owing to the decrease in the number of autopsies being performed. Inoculation of CMV into newborn mice, and extracorporeal preparations of mouse and guinea pig fetal inner ears, either in organ culture or as grafts on chick chorioallantoic membranes, yields viral perilabyrinthitis. The different ultrastructural appearances of CMV replicating in epithelial and mesenchymal cells show that animal CMV replicates in mesenchymal cells and human CMV replicates in epithelial cells of the inner ear. These different ultrastructural patterns indicate that the chromophobe (transitional) cells of the stria vascularis of the guinea pig are of mesenchymal origin. PMID- 6272580 TI - Skull base surgery. AB - Surgical inaccessibility, the obstacles of vital neural and vascular anatomy, and the overwhelming surgical mortality rate as a result of hemorrhage and sepsis have led surgeons to approach lesions of the skull base with understandable reluctance. We have, however, undergone a technical revolution in microsurgery, anesthesia, and neurodiagnosis. Innovative surgical minds have thus been armed with the technology to make surgery a reasonable alternative for these dreaded lesions. This article singles out the glomus tumor and its associated lesions, which the neuro-otologist must approach transtemporally. Their diagnosis and new treatment concepts are discussed. Old criteria for unresectability are redefined and new classifications are established. Surgery of these skull base lesions is discussed from the standpoint of the basic principles of exposure, hemostasis, and management of the facial nerve. Problems in rehabilitation of postoperative deficits are discussed. PMID- 6272581 TI - Turnover and regulation of Na-K-ATPase in HeLa cells. AB - HeLa cells in log growth have 10(6) surface Na-K-ATPase molecules as estimated by the specific binding of [3H]-ouabain. Studies utilizing ouabain as a label show that the ligand is internalized at a rate corresponding to the turnover of three sets of Na-K-ATPase enzymes per generation. The label is taken up exclusively into a particulate cell compartment where it is codistributed with beta hexosaminidase, identifying the internal compartment as lysosomal. Turnover is an important parameter in the recovery of the cells from glycoside intoxication. The unmetabolized glycoside is subsequently released by exocytosis. 13C-density labeled Na-K-ATPase has been identified by specific phosphorylation of its catalytic subunit with [32P]ATP or [33P]ATP, and the rate of turnover of the density label is shown to be the same as the internalization of the ouabain labeled site. There is a transit time of about 4 h from the onset of synthesis of the catalytic subunit to its insertion in the surface membrane; 2,800 catalytic subunits are synthesized per minute per cell, and 2,100 are turned over K+ starved cells respond to the stress in 24-30 h with modulation of the surface density of Na-K-ATPase the synthetic rate remains constant; the number of functional enzymes per cell is controlled by change in the rate constant for turnover. PMID- 6272582 TI - Specific binding of nerve growth factor to normal and denervated canine heart. AB - Preparations of membranes from normal and surgically denervated canine heart were tested for binding of radiolabeled nerve growth factor. Specific binding was detected in both normal and denervated hearts. Binding was nonsaturable and complex for normal atria and ventricles and denervated atria but appeared to be saturable for denervated ventricles. Nerve growth factor bound per microgram of protein was lower in denervated ventricles than normal ventricles, whereas slightly higher binding to denervated atria than normal atria was observed. This binding could not be displaced by cytochrome c, insulin, or epidermal growth factor. The data indicate that a large part of binding to normal ventricles could be due to nerve terminals attached to the heart, but specific binding detected in the denervated ventricles may be an intrinsic property of the tissue itself. These sites may serve as storage or uptake sites to direct sympathetic innervation in the developing or reinnervating myocardium. PMID- 6272583 TI - Interaction of NH4+ and Li+ with the renal microvillus membrane Na+-H+ exchanger. AB - We have examined the interaction of NH4+ and Li+ with the Na+-H+ exchanger in rabbit renal microvillus membrane vesicles. Alkalinization of the intravesicular space, monitored with [14C]DMO uptake by flow dialysis, resulted from the addition of Cl- salts of Na+, Li+, or NH4+ to the external medium. The 22Na+ uptake rate, assayed by a rapid filtration technique, was saturable with respect to external Na+ concentration (KT 6 mM Na+) and was competitively inhibited by external Li+ (Ki 1.9 mM) and NH4+ (Ki 4.3 mM). Efflux by 22Na+ from vesicles preloaded with Na+ was stimulated by external Na+ and NH4+ but inhibited by external Li+. We conclude that the renal microvillus membrane Na+-H+ exchanger has affinity for Li+ and NH4+ and can mediate the exchange not only of Na+ for H+ but also of Na+ for Na+, Na+ for Nh4+, Li+ for H+, possibly NH4+ for H+, and perhaps Li+ for Na+. The physiological significance of exchange modes other than Na+-H+ exchange is not certain at present, but Na+-NH4+ exchange could play a role in the proximal tubular acidification process. PMID- 6272584 TI - Involvement of extracellular calcium in gastric stimulation. AB - We have tested whether external Ca2+ is required for either initiation or maintenance of secretory parameters, including membrane elaboration of oxyntic cells, in frog gastric mucosa. Ca2+ was removed from in vitro mucosal preparations [by washing repeatedly in Ca2+-free Ringer solution and adding 0.1 mM ethylene glycol-bis(beta-aminoethylether)-N,N'-tetraacetic acid to the serosal solution] either before (i.e., resting tissues) or after addition of stimulants. Electrophysiology [transepithelial potential difference (PD) and resistance], morphology (morphometric analysis of transmission electron micrographs), and transport (H+ secretion) were monitored. La3+ (1 mM) was added to the mucosal solution to help maintain resistance and PD. La3+ decreased tissue shunt conductance during Ca2+-free conditions, as evidenced by a decreased mucosal serosal flux of 22Na+, presumably by preserving tight-junction integrity. Secretion was elicited by histamine alone or in combination with dibutyryl cAMP and isobutylmethylxanthine (a phosphodiesterase inhibitor). External Ca2+ is not required for the initiation of H+ secretion or the accompanying morphological changes when the combined stimulants are used, whereas H+ secretion and the morphological change showed some Ca2+ dependency when histamine alone was used. Thus, histamine-elicited secretion seems to be more sensitive to Ca2+ removal than that brought about by the combined stimulants. Long-term effects of Ca2+ free solutions on resistance, PD, and H+ secretion can largely be explained by disruptive effects on tight junctions. PMID- 6272585 TI - Cation effects on acid secretion in rabbit gastric glands. AB - The ability of isolated gastric glands to produce acid as a function of intra- and extracellular concentrations of K+ and Na+ was investigated. Ouabain inhibited acid formation as measured by the aminopyrine (AP) accumulation technique in a dose-dependent manner with an ED50 of 2 X 10(-6) M at 5.4 mM extracellular K+. This inhibition was counteracted by increasing extracellular K+ or decreasing extracellular Na+. In K+-free solutions with regular Na+, the AP accumulation was almost totally abolished; the readdition of K+ rapidly restored the response with a K0.5 of 1 mM extracellular K+. In the absence of Na+, with or without ouabain, there was always a significant residual AP accumulation, but the K0.5 of extracellular K+ required for acid formation increased to about 20 mM. Despite repeated washings in Na+-K+-free solutions, the intracellular K+ content could not be decreased below 24 mM above the apparent K0.5 for K+. It was found that the intracellular K+ could be depleted without disturbance of the acid secretory function if the glands were treated with the neutral ionophore amphotericin B and ouabain in K+-Na+-free solutions. Complete dose-response curves of H+ secretion as a function of K+ concentration could now be obtained. Thus the K0.5 for K+ activation of AP accumulation was 16.5 +/- 0.9 mM; upon histamine stimulation of the glands, the K0.5 decreased to 10.4 +/- 0.7 mM. Intracellular K+ concentrations above approximately 60 mM inhibited AP accumulation. Na+ dose dependently inhibited the K+-induced response. At low extracellular K+ concentrations, a ouabain-insensitive K+ accumulation mechanism was unmasked. This K+ uptake was partly inhibited y p-chloromercuribenzene sulfonic acid and by inhibitors of the gastric H+-K+-ATPase and amplified by inhibitors of K+ conductance channels such as Ba2+. The K+ content of gastric glands is mainly regulated by the Na+-K+-ATPase, but additional K+-uptake pathways are present. The affinity for K+ is increased in histamine-stimulated glands, which might indicate one mechanism of stimulating the secretory response. Na+ inhibits secretion possibly by binding to the cytosolic cation site of the gastric ATPase. The cation relationship to acid secretion in the rabbit parietal cell is similar to that described for vesicles isolated from hog gastric mucosa. PMID- 6272586 TI - Intramitochondrial adenine nucleotides and energy-linked functions of heart mitochondria. AB - Isolated rabbit heart mitochondria were incubated with varying amounts of inorganic pyrophosphate in 250 mM sucrose to specifically decrease the pool size of endogenous adenine nucleotides. The endogenous adenine nucleotide content decreased by as much as 80% as a result of this treatment. Phosphorylating respiration (state 3) declined from about 340 to 180 nAtoms O . min-1 . mg protein-1 over the full range of intramitochondrial adenine nucleotides measured (approx 7.5-1.5 nmol/mg protein). Uncoupled and nonphosphorylating (state 4) rates of respiration were not greatly affected by adenine nucleotide depletion. Respiratory activity of the adenine nucleotide-depleted mitochondria was enhanced by addition of exogenous adenosine 5'-triphosphate (ATP). Partial depletion (approx 40%) of the intramitochondrial adenine nucleotides resulted in an impaired ability of heart mitochondria to retain Ca2+. Premature Ca2+ efflux was associated with organelle swelling and altered energy coupling. Exogenous ATP or adenosine 5'-diphosphate (ADP) added prior to Ca2+ efflux restored Ca2+ retention in these mitochondria. Atractyloside inhibited the restoration of Ca2+ retention. This study indicates a significant role for endogenous adenine nucleotides in maintaining oxidative phosphorylation and Ca2+ transport in heart mitochondria. The results are discussed with regard to significance in ischemic heart damage. PMID- 6272587 TI - Antidepressants and alpha 2-adrenergic responsivity. PMID- 6272588 TI - [Hypophyseal adrenocorticotropic function and adrenal cortical function in women following labor complicated by massive blood loss]. PMID- 6272589 TI - [Functional characteristics of the corpus luteum]. PMID- 6272590 TI - [Use of a quantification scale for assessing the degree of severity of the climacteric syndrome and its treatment effectiveness]. PMID- 6272591 TI - [Herpetic diseases in women and newborn infants]. PMID- 6272592 TI - Origins of cancer in the neonate. PMID- 6272593 TI - Altered invertebrate nerve function after chronic exposure to low concentrations of alcohol. AB - Previous studies of direct alcohol toxicity on nerve tissue have been carried out using acute, extremely high doses of alcohol. Chronic administration of 20 mM ethanol to the mollusc Aplysia californica was achieved by adding ethanol to surrounding seawater. Although the animals appeared healthy, isolated ganglion cells from treated animals had significantly decreased mean action potential amplitudes and prolonged mean action potential durations compared to controls. These findings suggest that chronic exposure to a low concentration of ethanol comparable to that producing drunkeness in humans may have a direct toxic effect on invertebrate nerve tissue. PMID- 6272594 TI - [The effect of enflurane anaesthesia and operation on the sympathoadrenal and adrenocortical system (author's transl)]. AB - The effect of enflurane anaesthesia on the sympathoadrenal system, the hypothalamic-hypophyseal-adrenocortical system, and on the circulatory system was investigated in patients undergoing ophthalmic surgery. Blood concentrations of adrenaline, noradrenaline, ACTH, cortisol, and arterial blood pressure and heart rate were measured in 10 patients 30 min after premedication (I), 3-5 min after surgical incision (II), 20 or 45 min after surgical incision (III) and in 7 of 10 patients 10-15 min after extubation (IV). After conventional induction anaesthesia was maintained with enflurane (1.5-2.0%), combined with N2O/O2 2:1, and intermittent relaxation. Plasma adrenaline, ACTH, cortisol and heart rate increased significantly during operation, while plasma noradrenaline and blood pressure did not change significantly. Thus enflurane anaesthesia could not completely inhibit the increased activity of sympathoadrenal and hypothalamic hypophyseal-adrenocortical system caused by surgical stress. PMID- 6272595 TI - Murine leukemic lymphoblasts: homogenization and fractionation procedures for plasma membrane vesicles isolation. PMID- 6272596 TI - Purification of supercoiled DNA of plasmid col E1 by RPC-5 chromatography. PMID- 6272597 TI - A sensitive radioenzymatic assay for ATP. PMID- 6272598 TI - A sensitive assay for proteins and biliproteins. PMID- 6272599 TI - High-performance gel permeation chromatography of proteins in denaturing solvents and its application to the analysis of enveloped virus polypeptides. PMID- 6272600 TI - Improved estimation of DNA fragment lengths from Agarose gels. PMID- 6272602 TI - The effect of spermidine on endonuclease inhibition by agarose contaminants. PMID- 6272601 TI - A sensitive collagenase assay using [3H] collagen labeled by reaction with pyridoxal phosphate and [3H] borohydride. PMID- 6272603 TI - Fluorimetric microassay of DNA using a modified thiobarbituric acid assay. PMID- 6272604 TI - Determination of diadenosine tetraphosphate (Ap4A) levels in subpicomole quantities by a phosphodiesterase luciferin--luciferase coupled assay: application as a specific assay for diadenosine tetraphosphatase. PMID- 6272605 TI - 3-([3-125I]Iodo-4-hydroxyphenyl)propionyl carbohydrazide, a new radioiodination reagent for ultrasensitive detection and determination of periodate-oxidized nucleoside derivatives and other carbonyl compounds. PMID- 6272606 TI - Rapid analysis of small nucleic acid samples by gel electrophoresis. PMID- 6272607 TI - Rapid and efficient method for analyzing phosphorylation of the S6 ribosomal protein in 32Pi-labeled, tissue culture cells. PMID- 6272608 TI - A spectrophotometric procedure for the determination of activity of restriction endonucleases. PMID- 6272609 TI - Protonometry: the measurement of reaction velocities by changes in proton concentration. PMID- 6272610 TI - Use of solubilizable acrylamide disulfide gels for isolation of DNA fragments suitable for sequence analysis. PMID- 6272611 TI - Quantitation of vitamin D and its metabolites and their plasma concentrations in five species of animals. PMID- 6272612 TI - An improved radioreceptor assay for 1,25-dihydroxyvitamin D in human plasma. PMID- 6272613 TI - A fluorometric-high-performance liquid chromatographic assay procedure for several enzymatic activities using formycin analogs of adenosine 5'-mono, 5'-tri, and cyclic 3',5'-monophosphate as substrates. PMID- 6272614 TI - A method for determining DNA and chondrocyte content of articular cartilage. PMID- 6272615 TI - Ultrastructural analyses of control and enzyme-treated isolated renal basement membranes. PMID- 6272616 TI - Morphological correlates of hormone secretion in the rat adrenal cortex and the role of filopodia. AB - Rat adrenals in different states of stimulation were examined by transmission electron microscopy following perfusion fixation using an in situ isolated circulation technique. In unstimulated glands, intracortical capillaries were constricted and the cells of the cortex were pressed closely together with little development of filopodia or intercellular spaces. Glands fixed during the period of operative stress, or following a 1 hr perfusion with Adrenocorticotropic hormone (ACTH) showed that the radially orientated capillaries of the cortex were massively expanded, and the cells of both the glomerulosa and fasciculata exhibited an extensive development of filopodia on their surfaces. These filopodia extended into large intercellular spaces, where they often entered into complex relationships with filopodia from neighboring cells. The development of filopodia by cells of the adrenal cortex was also observed using scanning electron microscope techniques. In cells either incubated with ACTH in vitro or isolated from adrenals of rats treated with ACTH in vivo, the filopodia were numerous, often branched, and could reach as much as 1 micro m in length. In contrast, adrenal cells obtained from animals pretreated with cortisol were smooth surfaced. Other cell characteristics, including mitochondria, smooth endoplasmic reticulum, dense granules, and coated vesicles did not show such dramatic correlations with the state of stimulation. It is considered that the development of filopodia and intercellular space is related to secretory mechanisms in the rat adrenal cortex. PMID- 6272617 TI - Immune-enhanced phagocytic dysfunction in pulmonary macrophages infected with parainfluenza 1 (Sendai) virus. AB - Cultured alveolar macrophages infected with parainfluenza 1 (Sendai) virus were treated with specific antiviral immune serum and their phagocytic activity for opsonized erythrocytes (EA), Candida krusei, and Staphylococcus epidermidis quantitated. Membrane Fc receptor and candida binding activity were unaffected by the viral infection. In contrast, the virus infection decreased the phagocytic ingestion of EA. The addition of immune serum induced new phagocytic defects in that the treatment of virus-infected macrophages decreased the binding of EA and candida and reduced the ingestion of the yeast and the staphylococci. In addition, treatment with immune serum also enhanced the phagocytic defects induced by the virus infection alone, further reducing the binding and ingestion of EA. Neither virus infection nor treatment with immune serum affected the intracellular killing of S. epidermidis. These data demonstrated that virus infection of alveolar macrophages in vitro induce phagocytic defects that are accentuated by the treatment of the macrophages with antiviral antibody. PMID- 6272618 TI - Cells originating from sarcoid granulomas in vitro. AB - In an attempt to obtain in vitro experimental models for sarcoidosis, the primary cultures of granuloma cells were initiated with lymph nodes from 10 sarcoidosis patients. The cells migrating from tissue explants became confluent at 2 wk of culture. These cells exhibited overall morphologic features and enzymatic activities resembling those of in vivo granuloma epithelioid cells. By light and electron microscopy, the epithelioid cell in vitro contained a clear, large nucleus with 1 or 2 prominent nucleoli. The cytoplasm was characterized by the occurrence of lysosomal dense bodies and electron-lucent vacuoles, besides many mitochondria and well-developed Golgi complexes. The cell surface exhibited many processes, mainly lamellipodia. Large amounts of angiotensin converting enzyme activity (3.9 to 50.1 nmol/min/ml) lysozyme activity (3.9 to 23.0 micrograms/ml) were demonstrated in the medium used for culture. The results indicated that the major cell population grown in these cultures is derived from the epithelioid cells in sarcoid granulomas. PMID- 6272619 TI - Surgical ovarian lesions in children. PMID- 6272620 TI - Peliosis hepatis associated with cavernous hemangioma and hepatocarcinoma. AB - Peliosis hepatis is a rare lesion characterized by blood-filled cavities randomly distributed in the liver. It has been associated with the administration of medications such androgen-anabolic steroids, estrogens, oral contraceptives, and azathioprine. Its diagnosis is most reliably obtained by arteriogram and liver biopsy. Treatment consists of the discontinuance of those medications known to induce liver changes. Surgical treatment is reserved for those cases where a concomitant neoplasm or aneurysm is found. PMID- 6272622 TI - [Waldenstrom's macroglobulinemia and peripheral amyloid neuropathy (author's transl)]. AB - A 77-year-old man was diagnosed as having Waldenstrom's macroglobulinemia sex years after developing a sensory-motor neuropathy of the four limbs. The amyloid nature of the neuropathy was confirmed after biopsy of a peripheral nerve. The pathogenesis of the amyloidosis occurring during the course of dysglobulinemias is discussed, and those dysglobulinemias associated with non-amyloid neuropathies briefly described. PMID- 6272621 TI - Diphtheria, tetanus, and pertussis: Guidelines for vaccine prophylaxis and other preventive measures. Immunization Practices Advisory Committee, Centers for Disease Control. PMID- 6272624 TI - Virus-induced electrotonic coupling: hypothesis on the mechanism of periodic EEG discharges in Creutzfeldt-Jakob disease. AB - Experimental evidence and computer modeling indicate that periodic synchronous cellular depolarizing bursts (interictal spikes) arise when the balance recurrent inhibition and local excitatory coupling is altered. Such a mechanism may explain the generalized periodic sharp waves that characterize the electroencephalogram of many patients with Creutzfeldt-Jakob disease. In Creutzfeldt-Jakob disease, fusions of neuronal processes, particularly dendrites, may lead to abnormal electrotonic coupling between cells, providing the basis for powerful excitatory interaction whereby large neuronal aggregates burst in near synchrony. Cortical synchronous discharges would give rise to sharp waves in the electroencephalogram, whereas similar discharges in brainstem, spinal cord, or elsewhere could lead to myoclonic jerks. PMID- 6272623 TI - Family studies on nucleoside phosphorylase and the short arm of chromosome 14. AB - A family with two nucleoside phosphorylase-deficient patients has been scored for the segregation of NP0 and the variable region 14p. The mose likely 14p:NP recombination fraction is 0.15 in males and 0.30 in females. There is no family data to assign the Pi:Gm linkage group to chromosome 14, but as immunoglobulin heavy chain has been assigned to this chromosome by somatic cell methods the most likely gene order is 14p:NP:Pi:Gm with Pi in 14q2 and Gm in 14(q23 leads to q32), but the order 14p:NP:Gm:Pi with Pi in 14(q24 leads to qter) and Gm in 14(q22 leads to q24) is not excluded. The available linkage data between biochemical markers on acrocentric chromosomes and their short arm markers suggest that there may be more recombination towards the ends of human chromosomes whether or not those ends carry centromeres. PMID- 6272625 TI - Eaton-Lambert myasthenic syndrome: long-term treatment of three patients with prednisone. AB - Three patients with the myasthenic syndrome of Eaton-Lambert (ELS) were treated with prednisone for 6 and 12 months and more than 3 years, respectively. Muscle strength increased considerably in all, reaching a peak after 3 to 4 months, and was accompanied by simultaneous electromyographic improvement. Reduction of prednisone led to deterioration in all 3 patients; strength was regained when the higher dosage of prednisone was resumed. Prednisone is an alternative form of treatment for ELS, although the mechanism for its action is unclear. PMID- 6272626 TI - [Effect of ristomycin on the electrokinetic properties of Staphylococcus aureus cells]. AB - The electrophoretic mobility of the cells of Staph. aureus cultivated on the medium with ristomycin is markedly decreased at pH 3.0-5.0. This indicates that ristomycin added to the cultivation medium lowers the number of the phosphate groups in teichoic acids of the staphylococcal cell wall. The effect of ristomycin on the cells of the staphylococci was studied in vitro during the process of their incubation. It was shown that almost within the first minutes of the incubation ristomycin interacts immediately with the ionogenic groups of the cell wall and first of all with the phosphate groups of teichoic acids. PMID- 6272627 TI - Activity of trifluorothymidine against cytomegalovirus. AB - Trifluorothymidine (TFT) was tested for antiviral activity against mouse cytomegalovirus (MCMV) and human cytomegalovirus (HCMV) in one-step replication assays. The TFT concentration required to reduce virus yield by 50% (ID50) was 0.22 microM for MCMV and 0.012 microM for HCMV. The antiviral activity of TFT against MCMV was reversed by addition of equimolar thymidine, and no antiviral activity was demonstrable in a host cell line lacking thymidine kinase. Thus, TFT's anti-MCMV activity is dependent on a host cell TK since this herpesvirus lacks thymidine kinase. A continuous subcutaneous infusion of TFT achieving a serum concentration of 1 microM failed to protect mice from lethal MCMV infection, perhaps because serum levels of thymidine were comparable to the drug level. Comparison of the ID50 against HCMV and the ID50 against human bone marrow progenitor cells resulted in an in vitro therapeutic ratio of 108, suggesting that TFT might offer some promise as a clinically useful anti-HCMV agent. PMID- 6272628 TI - 6 beta-Iodopenicillanic acid (UI-38,006), a beta-lactamase inhibitor that extends the antibacterial spectrum of beta-lactam compounds: initial bacteriological characterization. AB - UK-38,006, 6 beta-iodopenicillanic acid, was shown to be a potent inhibitor of beta-lactamase enzymes. It potentiated the antibacterial action of ampicillin in vitro against beta-lactamase-producing strains of Staphylococcus aureus, Haemophilus influenzae, Bacteroides fragilis. Neisseria gonorrhoeae, and many Enterobacteriaceae. This ability to synergize with ampicillin was also demonstrated in vivo after oral administration of UK-38,006 to experimentally infected mice. UK-38,006 was also shown to synergize in vitro with other penicillins and cephalosporins against beta-lactamase-producing strains of Escherichia coli, Proteus mirabilis, and Klebsiella species. PMID- 6272629 TI - In vitro activity of ceftizoxime against Bacteroides fragilis: comparison with benzylpenicillin, cephalothin, and cefoxitin. AB - Minimum inhibitory concentrations of ceftizoxime for seven clinical isolates of Bacteroides fragilis were found to be markedly affected by inoculum size; the very low minimum inhibitory concentration values (less than or equal to 0.25 microgram/ml) obtained in tests with most strains were not sustained when the inoculum was raised 100-fold. Benzylpenicillin and cephalothin were also affected by inoculum size, but cefoxitin was not. Antibiotic assays showed that the strains inactivated ceftizoxime and cephalothin completely, benzylpenicillin partially, and cefoxitin not at all. Morphological investigations revealed that populations of B. fragilis responded differently to each of the four beta-lactam agents during the early stages of the encounter between B. fragilis and the antibiotic. Benzylpenicillin and cefoxitin induced spheroplast formation at lower concentrations than did ceftizoxime. Cephalothin evoked a novel morphological response that was not seen with other beta-lactam agents. PMID- 6272630 TI - Aminoglycoside phosphotransferase-II-mediated amikacin resistance in Escherichia coli. AB - An Escherichia coli strain with a plasmidic amikacin resistance has been selected for which the evidence strongly indicates that resistance is mediated by aminoglycoside phosphotransferase [APH(3')-II]: (i) this resistance was coupled with resistance against kanamycin and neomycin; (ii) partially purified APH(3') II[APH(3") free] modified amikacin by phosphorylation; (iii) the product of the APH(3')-II mediated reaction (i.e., 3'-O-phosphoryl-amikacin) lost its antibacterial activity; and (iv) the amikacin-modifying APH(3')-II activity increased 5- to 10-fold after adaptation of the cells to higher concentrations of amikacin. The substrate spectrum of this enzyme showed a low activity against amikacin as compared with neomycin. It is argued that the enzyme level rather than its substrate spectrum is important for enzyme-mediated resistance. The increase in enzyme levels was found to be correlated with an increase in copy number of a 110-Megadalton plasmid (pBN66) which coded for the APH(3')-II and the APH(3") activity. The increase in copy number was irreversible, and therefore this phenomenon is ascribed to a mutation of a gene which affects the copy number. In transconjugants, the original low copy number was present, and therefore the mutation must be located on the chromosome and not on the plasmid. PMID- 6272631 TI - Pharmacology of ceftizoxime compared with that of cefamandole. AB - The pharmacokinetics of ceftizoxime, a new beta-lactam antibiotic, were studied in normal, male volunteers and compared with the pharmacokinetics of cefamandole. After administration of 500 mg intramuscularly, ceftizoxime produced a peak level of 13.7 +/- 1 microgram/ml, compared with 13.2 +/- 1.6 microgram/ml for cefamandole. At 4 h, the serum level of ceftizoxime was 4.8 micrograms/ml, and that of cefamandole was 1.9 microgram/ml. At 8 h, ceftizoxime was still detected at 0.73 microgram/ml, whereas cefamandole was not. The half-life of ceftizoxime after intramuscular administration was 1.7 h, compared with 1 h for cefamandole. Serum levels of ceftizoxime and cefamandole after 1 g infused over 30 min were 84 and 88 micrograms/ml, respectively. At 5 h cefamandole was not detectable, whereas ceftizoxime had a serum level of 4.5 micrograms/ml and, at 7 h, 2.1 micrograms/ml. The half-life of ceftizoxime was 1.9 h, compared with 0.78 h for cefamandole. Urinary recovery of ceftizoxime after intramuscular and intravenous administration was 70 and 80%, respectively, compared with 78 and 73% for cefamandole. PMID- 6272632 TI - Chloramphenicol transposons found in Salmonella naestved and Escherichia coli of domestic animal origin. AB - Salmonella naestved strain AHI-21, of calf origin, harbors a conjugative R plasmid of group H1, pTE21, which encodes resistance to chloramphenicol (Cm), tetracycline, streptomycin, and sulfadimethoxine. Escherichia coli strain AHI-1, of pig origin, also harbors a conjugative R plasmid of group I alpha, pTE1, which encodes resistance to chloramphenicol and trimethoprim. When either of these R plasmids coexisted with a nonconjugative plasmid, pMK1, which is a composite plasmid of ColE1 and a kanamycin transposon (Tn5), transposition of the Cmr gene into pMK1 occurred independently of the host recA gene function, indicating that both R plasmids contained Cm transposons, Tn3351 and Tn3352. Electron microscopic analysis of self-annealed and heteroduplex molecules showed that they were of approximately 1.7 megadaltons in size and were inserted within the ColE1 loop region of pMK1. However, inverted repeat structures were not seen in these two Cm transposons. Restriction enzyme cleavage analysis showed that both Tn3351 and Tn3352 were indistinguishable in their cleavage patterns, suggesting that they were almost identical in deoxyribonucleic acid sequence despite the difference in their origin. These results suggest that the reciprocal transposition of the Cmr gene might have occurred between Salmonella and E. coli in nature. PMID- 6272633 TI - Metronidazole metabolism in cultures of Entamoeba histolytica and Trichomonas vaginalis. AB - Acetamide forms from metronidazole in cultures of either Entamoeba histolytica or Trichomonas vaginalis as it does in cultures of bacteria which are susceptible to this drug. Under aerobic conditions, the formation of acetamide is more rapid in a strain of T. vaginalis which is more susceptible to metronidazole. Thus, it appears that the formation of acetamide may be associated with the microbiocidal action of metronidazole. PMID- 6272634 TI - Differential activity of potential antiviral nucleoside analogs on herpes simplex virus-induced and human cellular thymidine kinases. AB - Potential antiviral nucleoside analogs 1-beta-D-arabinofuranosylthymine, the 1-(2 deoxy-2-fluoro-beta-D-arabinofuranosyl)-nucleosides of -5-methyluracil, -5 iodouracil, -5-methylcytosine, -5-iodocytosine, and -E-5-(2-bromovinyl)uracil, E 5-(2-bromovinyl)-2'-deoxyuridine, E-5-(2-bromovinyl)-1-beta-D arabinofuranosyluracil, and 9-(2-hydroxyethyoxymethyl)guanine were studied to compare their phosphorylation rates relative to thymidine by purified thymidine kinases from human and herpes simplex virus sources. Most of these analogs are capable of being phosphorylated by both human and viral enzymes. On the assumption that inhibition constants (Ki) reflect binding affinity, Ki values were determined for these analogs with the same thymidine kinases. In general, these analogs have a greater affinity for the viral enzymes. The amount of the analogs phosphorylated to the monophosphate form, which is presumably necessary to produce cytotoxic effects, was determined by the combined effects of phosphorylation rates and binding affinities. All of these analogs act as preferential substrates for the viral thymidine kinases at low concentrations, which may be one of the main reasons for their selective antiviral action. PMID- 6272635 TI - Studies on avian erythrocyte metabolism: purification and properties of myo inositol 1-phosphatase form chick erythrocytes. PMID- 6272636 TI - Studies on the immunological properties of complex V (mitochondrial ATP synthetase complex). PMID- 6272637 TI - Ca2+ transport against its electrochemical gradient in cytochrome oxidase vesicles reconstituted with mitochondrial hydrophobic proteins. PMID- 6272638 TI - Purification of galactose-1-phosphate uridylyltransferase from human placenta. PMID- 6272639 TI - The effect of phospholipid depletion on the EPR behavior of cytochrome oxidase. PMID- 6272640 TI - Phospholipids and regulation of protein kinase reaction. PMID- 6272641 TI - cAMP-dependent protein kinases, their subunits, and cAMP-binding protein from four sources: a comparison of physical characteristics determined by polyacrylamide gel electrophoresis. PMID- 6272642 TI - Magnetic resonance studies on interaction of bovine brain hexokinase with manganous ion, glucose, and glucose 6-phosphate. PMID- 6272643 TI - Correlation between phosphatidylcholine labeling and hormone receptor levels in alveolar type II epithelial cells: effects of dexamethasone and epidermal growth factor. PMID- 6272645 TI - Diffusion-controlled reaction kinetics of the binding of carbon monoxide to the heme undecapeptide of cytochrome c (microperoxidase 11) in high viscosity solvents. PMID- 6272644 TI - Human somatotropin: semisynthesis of the hormone by noncovalent interaction of the natural NH2-terminal fragment with a synthetic analog of the COOH-terminal fragment. PMID- 6272646 TI - S1 nuclease of Aspergillus oryzae: characterization of the associated phosphomonoesterase activity. PMID- 6272647 TI - Enzymes involved in the repair of damaged DNA. PMID- 6272648 TI - The p-nitrophenyl phosphatase activity of muscle carbonic anhydrase. PMID- 6272649 TI - Modification of (Na+ + K+)-dependent ATPase by fluorescein isothiocyanate: evidence for the involvement of different amino groups at different PH values. PMID- 6272650 TI - Influence of flavin addition and removal on the formation of superoxide by NADPH Cytochrome P-450 reductase: a spin-trap study. PMID- 6272651 TI - Characterization of N-alpha-acetylation of corticotropin fragments by a rat pituitary enzyme. PMID- 6272652 TI - The purification and properties of sucrose-phosphate synthetase from spinach leaves: the involvement of this enzyme and fructose bisphosphatase in the regulation of sucrose biosynthesis. PMID- 6272653 TI - Microsomal 17 beta-hydroxysteroid dehydrogenase of guinea pig liver: substrate and inhibitor specificity and effects of PH on steroid-enzyme interaction. PMID- 6272654 TI - The enthalpy changes upon hydrolysis of guanosine triphosphate anhydride and ester bonds. PMID- 6272655 TI - 5'-Nucleotidases of retinal rod membranes. PMID- 6272656 TI - Elemental content of vegetables and millet grown in potted soil amended with lignite fly ash. AB - Vegetables and millet were grown to maturity in potted soil amended with 10% (by weight) of lignite fly ash. Analysis of the ash, soil, and crops for 40 elements showed Al, As, B, Mg, Mo, Rb, and Se to be absorbed at higher levels by the ash grown plants than the controls in most instances. Selenium was of most concern, because of its toxicity and degree of plant uptake. PMID- 6272657 TI - Effect of morphine on diuresis in normal and diabetes insipidus dogs. PMID- 6272659 TI - Effect of a low protein diet on rats experimentally infected with Entamoeba histolytica. PMID- 6272660 TI - Migration of hamster sperm within capillaries: effect of agents elevating cyclic AMP. AB - Semiautomated capillary scanning was used to quantitate the migratory rate of hamster caudal epididymal (HCE) sperm populations from undiluted exudates into various defined media. The populations migrated through calcium-containing Tyrode's solution four times more rapidly than they did through buffered-glucose fortified saline or isotonic sucrose. This difference was partially eliminated by the addition to the saline or sucrose of the motility inducers, calcium ion or cyclic adenosine monophosphate (cAMP), and completely eliminated by the additional presence of the motility amplifiers, caffeine or spermine. The addition of the motility amplifiers, caffeine or spermine, alone to either calcium-saline or Tyrode's solution greatly stimulated the microscopically judged vigor of motility. However, this increase in flagellar activity was not coupled to increased forward velocity. Instead, as in the case of capacitated HCE sperm, the activation of motility resulted in significantly reduced forward velocities. Thus, it appears that under certain conditions caffeine, spermine, or capacitation can elevate sperm cAMP concentrations above those optimal for maximal forward progression. PMID- 6272661 TI - Vasoconstriction induced by delta 9-tetrahydrocannabinol on the perfused rabbit ear artery. AB - Periarterial incubation of delta 9-Tetrahydrocannabinol (THC, 15.0--30.0 mM) to the "in vitro" perfused central artery of the rabbit ear, induced sustained contractions related to the dose and reverted after washing. Simultaneously, the artery responses to transmural electrical stimulation were blocked and the effects of intravascular injection of noradrenaline were additive to the constriction induced by THC. Either reserpine treatment or chronic artery denervation blocked the vasoconstriction induced by THC. These results indicate that THC may induce vasoconstriction in the perfused artery of the rabbit through the release of noradrenaline from sympathetic nerve terminals. PMID- 6272662 TI - Unusual tubular inclusions in the mitochondrial matrix from human livers with cholelithiasis. AB - Unusual tubular inclusions in the mitochondrial matrix of human hepatocytes were found in 7 out of 15 cases with cholelithiasis. These inclusions were composed of tubular structures which were about 50 nm in diameter and varying in number. The wall of the individual tubules appeared to consist of about 10 tubular subunits with a diameter of about 5 nm. Although some relationship between occurrence of these tubular inclusions and production of gallstones might be suggested, their origin and functional significance are unknown. PMID- 6272658 TI - Heberden Oration 1980: aspects of the cell biology of the rheumatoid synovial lesion. PMID- 6272664 TI - Granular cell tumor of the esophagus. PMID- 6272663 TI - Changes in serum immunoglobulin concentrations in patients with multiple sclerosis during the treatment with ACTH. AB - Changes in serum immunoglobulin (IgA, IgG, IgM) concentrations were studied in 31 patients with multiple sclerosis (MS) according to the clinical form of the disease, its duration and treatment with ACTH. The serum IgG concentration was increased on the onset of the disease, while it was significantly decreased in MS patients treated with ACTH for the first time. There was no significant relationship of serum immunoglobulin concentrations and the number of courses of treatment with ACTH or duration of MS. PMID- 6272665 TI - Sural nerve conduction studies and late responses in children undergoing hemodialysis. AB - Physiologic evidence of peripheral neuropathy has been described previously in children undergoing hemodialysis. In order to detect early or subclinical evidence of peripheral neuropathy, several newer electrophysiologic techniques, including latencies of late responses (H reflex and F response) and sural nerve sensory studies, were evaluated in addition to routine motor and sensory conduction in 17 randomly selected children (mean age 14.2 years) undergoing hemodialysis (12 of whom had no clinical evidence of peripheral neuropathy) and 20 age-matched normal control subjects. Conventional motor and sensory conduction studies of median and ulnar nerves and motor conduction of peroneal and tibial nerves showed abnormalities of motor conduction in 5 (29%) and abnormalities of sensory conduction in 2 (12%). Sural nerve sensory potentials were abnormal in 10 (59%) patients. Late response were significantly abnormal in 10 (59%) patients, 5 (29%) of whom had normal routine motor conduction studies in the same nerve distribution. The abnormalities of late responses and motor and sensory conduction were more evident in lower limbs. Studies of late responses and sural sensory conduction provide a method of detecting subclinical neuropathy in this patient population at a time when results of conventional motor and sensory conduction tests are within normal limits. Effects of dialysis may then be followed quantitatively in patients whose neuropathy would otherwise be undetectable. PMID- 6272666 TI - Studies on the neutralizing antibody to herpes simplex virus. II. Quantitative analysis of type specific antibodies in rabbits immunized with both types of virus. AB - A quantitative analysis was carried out on type specific antibody developed in rabbits primed with three doses of one type of herpes simplex virus at two week intervals, followed by administration of another type of virus. Complement requiring neutralizing antibody assay and mutual absorbing methods were used. The homotype specific antibody produced against the type of virus used for priming persisted at a constant level for a long period, despite the lack of antigenic stimuli by the same type after priming. The homotype specific antibody was not affected by the heterotypic antigenic stimuli. On the other hand, the heterotype specific antibody developed after the first or second injection of heterotypic virus, and the titer tended to decline, reaching undetectable levels at 9 months. However, the heterotype specific antibody was produced rapidly after the third injection of virus at 9 months and the titers exceeded or closed to those of the homotype specific antibody. The homotype specific antibodies were mainly distributed in the IgG fraction, while the IgM fraction contained mainly the heterotype specific antibodies. In relationship between type specific antibody response and the values of II/I index, the coexistence of both type specific antibodies was apparent in sera with the values of 91 to 117 of II/I index and in the IgG fraction with the values of 90 to 115, respectively. PMID- 6272668 TI - Augmentation in the infectivity of SV40 virus DNA by amphotericin B methyl ester: brief report. PMID- 6272667 TI - Relation of Bovine leukosis virus production on cell growth cycle. AB - The cell cycle of fetal lamb kidney (FLK) cultures chronically infected with bovine leukosis virus was synchronized by double thymidine block. The synchronized FLK cells were examined by production of BLV antigen and virion release by cytoplasmic immunofluorescence and syncytia forming assay, respectively. The production of BLV antigens was increased during S and G2 phases and was decreased during M and G1 phases. BLV release was associated with the M phase of FLK cells. Short term lymphocyte cultures from BLV infected cattle were treated with hydroxyurea and mitomycin C. The expression of BLV antigen and DNA synthesis of PHA stimulated lymphocytes was inhibited by both drugs. PMID- 6272670 TI - An unusual case of encephalomyeloradiculoneuropathy in a young woman. AB - Fulminant and progressive CNS disease developed in a young woman and was associated with bilateral deafness and sharply elevated CSF protein levels. Two months after onset, a severe sensorimotor neuropathy developed. Clinical improvement occurred with corticosteroid therapy. PMID- 6272669 TI - [Schwannoma with granular cells. Indirect evidence of the closeness of schwannoma and Abrikosov's tumor]. AB - A woman of 29 developed a slowly growing subcutaneous tumor (1.5 X 1 X 0.5 cm) of the forehead. This neoplasm showed the histological pattern of neurilemmoma with the presence of dispersed rectangular cells having acidophilic fine granular cytoplasm imitating those of Abrikosov's tumor. This case as well as a few others described in the literature seem to confirm the thesis of neurilemmal origin of the so-called Abrikosov's myoblastoma. PMID- 6272671 TI - Characterization and relationship to exocrine secretion of rat parotid gland cyclic AMP-dependent protein kinase. PMID- 6272672 TI - Oxidative destruction of the microbial metabolite aflatoxin by the myeloperoxidase-hydrogen peroxide-chloride system. PMID- 6272673 TI - Endogenous Candida albicans endophthalmitis in the rabbit. Chemotherapy for systemic effect. AB - Progressive endogenous Candida albicans endophthalmitis was established in rabbits by intravenous (IV) injection of blastospores (2.0 to 5.0 x 10/kg). Severity of infection was directly related to the strain and inoculum size. Intravenous amphotericin B (1.0 mg/kg/day), IV amphotericin B methyl ester ascorbate (5.0 mg/kg/day), and oral ketoconazole (80 mg/kg/day) effectively prevented or reduced the severity of infection when therapy was initiated 24 hours following inoculation of blastospores and continued for five to seven days. Intravenous miconazole (30 mg/kg/day) was ineffective in this model. Intravenous amphotericin B(1.0 to 2.0 mg/kg on alternate days), IV amphotericin B methyl ester ascorbate (5.0 mg/kg/day), and oral ketoconazole (80 mg/kg/day reduced the severity of C albicans endophthalmitis when therapy was initiated seven days following injection of blastospores and continued for 28 days. Oral flucytosine (75 and 150 mg/kg/day in four doses) produced uniformly fatal hepatic necrosis in uninfected rabbits. PMID- 6272674 TI - Enzymes of galactose metabolism in livers of suckling and adult tammar wallabies (Macropus eugenii) and other marsupials. AB - The activities of galactokinase, hexose-1-phosphate uridylyl transferase and UDPglucose 4-epimerase in homogenates of livers of two adult and 20 suckling tammar wallabies aged from 6 to 50 weeks were investigated. The activities of all three enzymes were high until 24-30 weeks post partum, after which they declined to low levels. The activities of the three liver enzymes were high in pouch young of six other species of marsupial. Comparison of the activities of the three liver enzymes in suckling tammar wallabies with those in suckling rats showed no difference between the two species in regard to galactokinase and uridylyl transferase, but the UDPglucose 4-epimerase activity in tammar wallabies was approximately double than found in rats. This may be related to the high galactose content of tammar wallaby milk compared with rat milk. In suckling tammar wallabies, the liver had higher enzyme activities than other tissues studied. It is concluded that, contrary to the suggestion of Stephens et al. (1975), pouch young marsupials are not deficient in their ability to metabolize galactose. PMID- 6272675 TI - Modulation of functional capacity of small ovarian follicles in the post-partum cow by prolactin. AB - This study was undertaken to examine the possibility that the prolonged anovulatory period frequently experienced by the post-partum cow is due to a disruption of function at the ovarian level promoted by the high, suckling induced, blood prolactin concentrations. Fifteen cows, less than 35 days post partum, were allocated to three groups (1, 3 and 5) and given no hormonal treatment, prostaglandin plus pregnant mare serum gonadotrophin (PMSG) treatment or injected with 2-bromo-alpha-ergocryptine to reduce circulating prolactin levels. Ten synchronized cyclic cows were allocated to two groups (2 and 4) and given prostaglandin or prostaglandin plus PMGS treatment. All cows were ovariectomized 1 or 2 days after treatment of Graafian follicles less than 9 mm in diameter were selected after dissection from the ovaries. The follicles were cultured for 18 h with or without prolactin (1 microgram/ml) and steroid accumulation in the culture medium estimated. The follicles were then separated into theca and granulosa which were incubated for 40 min with LH (1 microgram/ml) or FSH (5 micrograms/ml). Cyclic AMP concentrations were estimated as an indication of tissue responsiveness to gonadotrophins. The secretion of oestradiol-17 beta, progesterone, testosterone or androstenedione during 18 h culture did not differ between follicles isolated from post-partum or cyclic cows. The presence of prolactin in the culture medium had no overall effect on steroid secretion although some specific effects within each group were noticed. Incubation with LH increased cyclic AMP levels in the theca but the granulosa did not respond. Likewise FSH increased cyclic AMP levels in granulosa preparations but not in theca. There were no differences in response between post-partum and cyclic cows, but exposure of the follicles to prolactin in vitro did significantly reduce the LH-induced increase in cyclic AMP levels in isolated theca. We have concluded that endogenous prolactin may modify but does not inhibit the resumption of ovarian function following parturition in the beef cow. PMID- 6272676 TI - Nitrofurantoin neuropathy. AB - Four cases of severe peripheral neuropathy directly attributable to nitrofurantoin are reported. The neuropathy was not dose-related and not necessarily associated with abnormal renal function. Recovery was slow, and neither severity nor recovery was related to the total dose of the drug. The pathological changes seen on light microscopy and electron microscopy were those of acute, severe axonal degeneration. It is emphasised that nitrofurantoin is a neurotic drug and should not be prescribed to the elderly not to anyone with impairment of renal function. PMID- 6272677 TI - ACTH-induced grooming involves high-affinity opiate receptors. PMID- 6272679 TI - Lithium chloride-produced prey aversion in the toad (Bufo americanus). PMID- 6272678 TI - Selective sensitization induced by lithium malaise and footshock in rats. PMID- 6272680 TI - Brief report food-aversion conditioning in Japanese monkeys (Macaca fuscata): a dissociation of feeding in two separate situations. PMID- 6272681 TI - Rapid communication recuperation from lithium-induced illness: flavor enhancement for rats. PMID- 6272682 TI - Segmental response to sympathetic activation in the canine coronary circulation. PMID- 6272683 TI - Constriction of the epicardial coronary artery induced by alpha-adrenergic stimulation. AB - Effects of alpha adrenergic stimulation on diameter change of the epicardial coronary artery were studied in in-situ canine heart. The diameter of the left circumflex coronary artery (LCX) was continuously measured by 10 MHz PZT crystals applied to the external wall of the LCX. Aortic pressure was measured using high fidelity catheter-tip manometer. Enhanced activity of alpha adrenergic receptor by humoral and/or somatic nerve stimulations caused a reduction of the LCX diameter despite of the systemic pressure rise. PMID- 6272684 TI - Regulation of large coronary vessels by adrenergic mechanisms in conscious dogs. AB - The effects of alpha and beta adrenergic stimulation were examined in conscious dogs on measurements of left circumflex coronary blood flow and coronary arterial diameter and on calculations of mean coronary resistance (MCR) and left circumflex coronary internal cross-sectional area (CSA). Methoxamine (50 micrograms/kg/min), after transiently increasing left circumflex coronary dimensions, induced sustained reductions in left circumflex coronary diameter (9 +/- 2%) and CSA (27 +/- 5%) at a time when mean arterial pressure rose by 65 +/- 5%, left ventricular (LV) dP/dt had decreased only slightly, and heart rate and mean coronary blood flow were at control levels. Isoproterenol, (0.1 microgram/kg/min), increased heart rate by 66 +/- 8%, LV dP/dt by 58 +/- 5%, and CSA by 17 +/- 3%, while it decreased mean arterial pressure by 12 +/- 1% and MCR by 44 +/- 5%. After beta 1 adrenergic receptor blockade and with heart rate held constant, isoproterenol did not increase LV dP/dt but decreased mean arterial pressure similarly (13 +/- 2%) and induced attenuated increases in CSA (6 +/- 1%), and decreases in MCR (17 +/- 3%). After combined beta 1 and beta 2 adrenergic receptor blockades isoproterenol induced no significant effects. Thus, in the conscious dog, large coronary vessels not only react passively to changes in aortic pressure but also undergo substantial active changes. Alpha adrenergic stimulation is sufficiently powerful to reduce CSA, despite the opposing elevation of distending pressure. Moreover, large vessels appear to be regulated by beta 1 adrenergic mediated increase in myocardial metabolic demands, as well as by beta 2 adrenergic medicated vasodilation. PMID- 6272685 TI - Norepinephrine and the coronary vascular bed in the conscious dog. PMID- 6272687 TI - Further studies on complementation between mutants of Clostridium perfringens. AB - 1. Mutants devoid of lambda- and kappa-toxin and hemagglutinin (HA), respectively, were isolated from Cl. perfringens PB6K. The lambda- and HA- mutants could be classified into a and b groups by complementation but the kappa- mutants were all of the a group. 2. All b group mutants isolated, irrespective of the marker used for isolation, were pleiotropically negative or leaky with respect to theta-, lambda- and kappa-toxin and HA production. 3. Lambda-toxin produced by complementation was proved to be a rennet-like protease. 4. The activities of 12 extracellular enzymes, including sialidase, of several b group strains and the parent PB6K were compared, but no definite differences were observed. From this finding, the productions of these enzymes were concluded not to be regulated by the same mechanism as theta-, lambda- and kappa-toxin and HA. 5. Cl. perfringens CN3870 was also studied. Findings were similar to those on PB6K except for very low activity of HA. PMID- 6272686 TI - Alpha-noradrenergic receptor binding sites in bovine aorta. PMID- 6272688 TI - Complementation characteristics of newly isolated mutants from two groups of strains of Clostridium perfringens. AB - 1. Mutants of a group were converted to b group by a second NG treatment. The resulting b group mutants could not produce the marker products that had been lost on the first NG treatment but could produce the others by complementation. 2. Mutants of b group were converted to constitutive mutants by a second NG treatment. No back mutation from b group was observed. PMID- 6272689 TI - Studies on the mechanism of antibody-mediated enhancement of Getah virus infectivity. AB - Inoculation on BHK-21 cells with Getah virus sensitized with hyperimmune homologous mouse antiserum resulted in higher infective titers than those obtained with non-sensitized control virus. This phenomenon was not observed with Vero cells. Experiments were carried out on the mechanism of this enhancement with the following results. When BHK-21 cells were pretreated with a mixture of UV-irradiated virus and antiserum before inoculation, the enhancement of infectivity of Getah virus was markedly decreased. IgG antibody against Getah virus which had been digested with 1--2% of pepsin did not show any enhancing activity. Sensitized sheep erythrocytes adhered to monolayered cultures of BHK-21 cells. These results indicate that the appearance of enhancing activity of a complex of the virus and antibody is closely related with the existence of a receptor for the Fc part of IgG on ordinary tissue culture cells, BHK-21 cells. PMID- 6272691 TI - Consistent appearance of microtubules in cells productively infected with various strains of type 2 herpes simplex virus. AB - Electron microscopic examination showed microtubular structures in FL cells infected with all 18 strains of HSV-2 examined, but not in cells infected with 9 strains of HSV-1. These structures were also detected in other cultured cells (Vero and Earle's L cells) infected with HSV-2, and also in vivo in cells, such as neuronal cells of the spinal ganglia and liver cells, of one-day-old suckling mice (DDD strain) infected with this type of virus. Thus the microtubules were consistently detected in cells productively infected with HSV-2. No other herpesviruses so far examined produced microtubular structures such as those observed in HSV-2 infected cells. PMID- 6272690 TI - Establishment of an attenuated ML-17 strain of Japanese encephalitis virus. PMID- 6272692 TI - Improved purification of brine-shrimp (Artemia saline) (Na+ + K+)-activated adenosine triphosphatase and amino-acid and carbohydrate analyses of the isolated subunits. AB - Purification of the (Na+ + K+)-activated ATPase has been improved 2-fold the respect to both purity and yield over the previous method [Peterson, Ewing, Hootman & Conte (1978) J. Biol. Chem. 253, 4762-4770] by using Lubrol WX and non denaturing concentrations of sodium dodecyl sulphate (SDS). The enzyme was purified 200-fold over the homogenate. The preparation had a specific activity of about 600 mumol of Pi/h per mg of protein, and was about 60% pure according to quantification of Coomassie Blue-stained SDS/polyacrylamide gels. The yield of purified enzyme was about 10 mg of protein per 100g of dry brine-shrimp (Artemia salina) cysts. The method is highly suitable for purification either on a small scale (10-25g of dry cysts) or on a large scale (900g of dry cysts) and methods are described for both. The large (Na+ + K+)-activated ATPase subunit (alpha subunit) was isolated in pure form by SDS-gel filtration on Bio-Gel A 1.5m. The small subunit (beta-subunit) was eluted with other contaminating proteins on the Bio-Gel column, but was isolated in pure form by extraction from SDS/polyacrylamide gels. The amino acid and carbohydrate compositions of both subunits are reported. The alpha-subunit contained 5.2% carbohydrate by weight, and the beta-subunit 9.2%. Sialic acid was absent from both subunits. PMID- 6272693 TI - The reconstitution of L-3-glycerophosphate-cytochrome c oxidoreductase from L-3 glycerophosphate dehydrogenase, ubiquinone-10 and ubiquinol-cytochrome c oxidoreductase. AB - Purified L-3-glycerophosphate dehydrogenase from pig brain mitochondria interacts with ubiquinone-10 and ubiquinol-cytochrome c oxidoreductase (Complex III) from bovine heart mitochondria to reconstitute antimycin-sensitive L-3 glycerophosphate- cytochrome c oxidoreductase. This activity is completely dependent on the two enzymes and largely dependent on ubiquinone-10. Reconstitution requires that the two enzymes should be simultaneously present in the same membranous aggregate produced by removal of detergent from the enzymes. Reconstitution by removing detergent by dialysis or dilution is inefficient because of self-aggregation of the dehydrogenase. Highly efficient reconstitution can be achieved if the enzymes are co-precipitated by addition of ethanol. The rate with reconstituted enzyme approaches that expected from the turnover of the dehydrogenase with ubiquinone-1 as acceptor. The behaviour of the reconstituted system shows some of the characteristics expected for a stoicheiometric association of one molecule of dehydrogenase with one molecule of Complex III. On raising the phospholipid/protein ratio, the dehydrogenase and Complex III appear to operate as independent enzymes acting in sequence. These effects are very similar to those observed for the interaction of NADH dehydrogenase and Complex III and are explained in terms of the model proposed by Heron, Ragan & Trumpower [(1978) biochem. J. 174, 791-800]. PMID- 6272695 TI - Antiserum against the catalytic subunit of adenosine 3':5'-cyclic monophosphate dependent protein kinase. Reactivity towards various protein kinases. AB - An antiserum against the catalytic subunit C of cyclic AMP-dependent protein kinase, isolated from bovine heart type II protein kinase, was produced in rabbits. Reaction of the catalytic subunit with antiserum and separation of the immunoglobulin G fraction by Protein A-Sepharose quantitatively removed the enzyme from solutions. Comparative immunotitration of protein kinases showed that the amount of antiserum required to eliminate 50% of the enzymic activity was identical for pure catalytic subunit, and for holoenzymes type I and type II. The reactivity of the holoenzymes with the antiserum was identical in the absence or the presence of dissociating concentrations of cyclic AMP. Most of the holoenzyme (type II) remains intact when bound to the antibodies as shown by quantification of the regulatory subunit in the supernatant of the immunoprecipitate. Titration with the antibodies also revealed the presence of a cyclic AMP-independent histone kinase in bovine heart protein kinase I preparations obtained by DEAE cellulose chromatography. Cyclic AMP-dependent protein kinase purified from the particulate fraction of bovine heart reacted with the antiserum to the same degree as the soluble enzyme, whereas two cyclic AMP-independent kinases separated from the particle fraction neither reacted with the antiserum nor influenced the reaction of the antibodies with the cyclic AMP-dependent protein kinase. Immunotitration of the protein kinase catalytic subunit C from rat liver revealed that the antibodies had rather similar reactivities towards the rat liver and the bovine heart enzyme. This points to a relatively high degree of homology of the catalytic subunit in mammalian tissues and species. Broad applicability of the antiserum to problems related to cyclic AMP-dependent protein kinases is thus indicated. PMID- 6272694 TI - The H+/e- stoicheiometry of respiration-linked proton translocation in the cytochrome system of mitochondria. AB - 1. The -->H(+)/e(-) quotients for proton release from mitochondria associated with electron flow from succinate and duroquinol to O(2), ferricyanide or ferricytochrome c, and from NNN'N'-tetramethyl-p-phenylenediamine+ascorbate to O(2), were determined from rate measurements of electron flow and proton translocation. 2. Care was taken to avoid, or to take into account, unrelated electron flow and proton translocation, which might take place in addition to the oxido-reductions that were the subject of our analysis. Spectrophotometric techniques were chosen to provide accurate measurement of the rate of consumption of oxidants and reductants. The rate of proton translocation was measured with fast pH meters with a precision of 10(-3) pH unit. 3. The -->H(+)/O quotient for succinate or duroquinol oxidation was, at neutral pH, 4, when computed on the basis of spectrophotometric determinations of the rate of O(2) consumption or duroquinol oxidation. Higher -->H(+)/O quotients for succinate oxidation, obtained from polarographic measurements of O(2) consumption, resulted from underestimation of the respiratory rate. 4. The -->H(+)/2e(-) quotient for electron flow from succinate and duroquinol to ferricyanide or ferricytochrome c ranged from 3.9 to 3.6. 5. Respiration elicited by NNN'N'-tetramethyl-p phenylenediamine+ascorbate by antimycin-inhibited mitochondria resulted in extra proton release in addition to that produced for oxidation of ascorbate to dehydroascorbate. Accurate spectrophotometric measurement of respiration showed that the -->H(+)/e(-) ratio was only 0.25 and not 0.7-1.0 as obtained with the inadequate polarographic assay of respiration. Proton release was practically suppressed when mitochondria were preincubated aerobically in the absence of antimycin. Furthermore, the rate of scalar proton consumption for water production was lower than that expected from the stoicheiometry. Thus the extra proton release observed during respiration elicited by NNN'N'-tetramethyl-p phenylenediamine+ascorbate is caused by oxidation of endogenous hydrogenated reductants. 6. It is concluded that (i) the -->H(+)/O quotient for the cytochrome system is, at neutral pH, 4 and not 6 or 8 as reported by others; (ii) all the four protons are released during electron flow from quinol to cytochrome c; (iii) the oxidase transfers electrons from cytochrome c to protons from the matrix aqueous phase and does not pump protons from the matrix to the outer aqueous phase. PMID- 6272696 TI - Theoretical analysis of the consequences of cyclic nucleotide phosphodiesterase negative co-operativity. Amplification and positive co-operativity of cyclic AMP accumulation. AB - Most tissues contain multiple forms of cyclic nucleotide phosphodiesterases (3':5'-cyclic-nucleotide 5' nucleotidohydrolase, EC 3.1.4.17). Consequently, in most, if not in all, tissues, substrate-velocity curves deviate from Michaelian kinetics and exhibit an apparent negative co-operativity. We have studied the possible theoretical consequences of this property on the quantitative features of cyclic AMP accumulation in response to activation of adenylate cyclase. Negative co-operativity of phosphodiesterases tends to generate a "positively co operative" cyclic AMP accumulation curve. It amplifies the stimulation of cyclic AMP accumulation as compared with the stimulation of cyclic AMP synthesis. It enhances the sensitivity of cyclic AMP accumulation to slight variation of phosphodiesterase maximal velocity. It tends to shift the cyclic AMP accumulation curve to higher concentrations of stimulator as compared with the adenylate cyclase activation curve. This accounts for much of the data in the literature of hormonal effects on phosphodiesterase activity. It shows that the characteristics of cyclic nucleotide phosphodiesterases are as important as those of adenylate cyclase in determining the response of the system. PMID- 6272697 TI - Changes in cyclic AMP and cyclic GMP concentrations during the action of 5 hydroxytryptamine on an insect salivary gland. AB - Salivary glands from adult blowflies (Calliphora erythrocephala Meigen) were studied in vitro. The time course of changes in cyclic AMP content of the glands was followed at different concentration of 5-hydroxytryptamine. There was an immediate biphasic rise and fall in cyclic AMP content, following by a slower rise and subsequent gradual decline. The initial rise preceded the onset of fluid secretion by the glands. Rises in cyclic AMP content were inhibited by compound RMI 12330 A (an adenylate cyclase inhibitor) and were halted after about 15-20s if the glands were deprived of Ca2+. Theophylline (a phosphodiesterase inhibitor) abolished the decline phase of the fast response, Losses of cyclic AMP from the glands either to the bathing medium or to the saliva were small and could not account for the rapid fall found. Evidence is presented that cyclic GMP is not involved in the process of initiating secretion in the blowfly salivary gland. PMID- 6272698 TI - Study of the fructose 6-phosphate/fructose 1,6-bi-phosphate cycle in the liver in vivo. AB - 1. The method proposed by Rognstad & Katz [(1976) Arch, Biochem, Biophys, 177, 337-345] for the determination of the fructose 6-phosphate/fructose 1,6 bisphosphate cycle by the randomization of carbon between C-1 and C-6 of glucose glucose formed from [1-14C] galactose was applied to anaesthetized rats and conscious mice. 2. It was checked that the hydrolysis of fructose 6-phosphate by glucose 6-phosphatase is too weak to invalidate the method. The participation of the Cori cycle in the randomization was negligible within the short experimental period used (2-4 min). 3. No detectable randomization of carbon was observed in starved animals, indicating that phosphofructokinase is inactive in this experimental condition. 4. Randomization of carbon was detected as soon as 1 min after administration of [1-14C] galactose to fed animals and was maximal at about 3-4 min. It was calculated that on average 15% of the glucose formed by the liver to fed rats was recycled through the triose phosphates. The extent of cycling was quite variable. Recycling was also observed in starved rats in which glucose had been administered intravenously 10 min previously. In these animals, recycling was completely inhibited by glucagon. 5. The main factors that appear to be responsible for the very large changes in recycling observed in various experimental conditions are the concentrations of fructose 1,6-bisphosphate and of fructose 6-phosphate and also the affinity of phosphofructokinase for fructose 6-phosphate. The concentration of nucleotides does not seem to play a role. PMID- 6272700 TI - Phosphatidylinositol phosphodiesterase in higher plants. AB - 1. The lower regions of the stem of celery (Apium graveolens L.) contain a soluble enzyme that hydrolyses phosphatidylinositol. 2. The lipoidal product of hydrolysis is diacylglycerol, and the water-soluble products are 1:2-cyclic phosphoinositol and phosphoinositol in the approximate proportions of 60% and 40% respectively: this indicates that a phosphodiesterase (phospholipase C-like) activity is cleaving the phosphatidylinositol. 3. The enzyme requires a bivalent cation, Ca2+ being the most effective activator. 4. The enzyme has a pH optimum, depending on conditions of assay, of pH 5.9-6.6 and in this pH range shows no detectable activity against phosphatidylcholine or phosphatidylethanolamine. 5. The activity is stimulated by phosphatidic acid and slightly inhibited (30% at concentrations equimolar with phosphatidylinositol) by phosphatidylcholine. 6. The phosphodiesterase was also detected (but not quantified) in the tips of the flowers in cauliflowers, in outer leaves of onion and in the elongating stem of daffodils. 7. The enzyme's properties are compared with equivalent mammalian enzymes, and its possible role in the catabolism of phosphatidylinositol in higher plants is discussed. PMID- 6272699 TI - Stoichiometry of catecholamine/proton exchange across the chromaffin-granule membrane. AB - Catecholamines are accumulated by bovine chromaffin-granule "ghosts" in the presence of MgATP at 25 degrees C. With low concentrations of catecholamine, ratios of internal to external amine concentration of up to 20 000 were obtained. These values fit well with a transport model in which amine accumulation is both electrogenic and dependent on a pH gradient across the membrane. PMID- 6272701 TI - Androgens regulate mitochondrial cytochrome c oxidase and lysosomal hydrolases in mouse skeletal muscle. AB - The gastrocnemius, a fast-twitch white muscle, and the soleus, a slow-twitch red muscle, were studied in A/J mice. The specific activities of the lysosomal hydrolases, beta-D-glucuronidase, hexosaminidase, beta-D-galactosidase and arylsulphatase, the inner-mitochondrial-membrane enzyme cytochrome c oxidase, and the outer-mitochondrial-membrane enzyme monoamine oxidase, were greater in the soleus than in the gastrocnemius. The specific activities of the lysosomal hydrolases and cytochrome c oxidase in the gastrocnemius and soleus were substantially higher in male mice than in female mice. Orchiectomy abolished this sex difference. Testosterone increased the activities of the lysosomal hydrolases and cytochrome c oxidase and coincidentally induced muscle hypertrophy and an accretion of protein and RNA, but total DNA remained constant. Monoamine oxidase was unaffected by sex, orchiectomy and testosterone. These findings indicate that endogenous androgens regulate the activity of enzymes associated with lysosomes and the inner mitochondrial membrane, as well as muscle fibre growth in mouse skeletal muscle. PMID- 6272702 TI - Inhibition of bone resorption in culture by inhibitors of thiol proteinases. AB - Leupeptin, antipain, tosyl-lysylchloromethane (Tos-Lys-CH2Cl) and benzyloxy carbonylphenylalanylalanyldiazomethane (Z-Phe-Ala-CHN2) inhibit reversibly the resorption induced by parathyroid hormone or heparin in cultured mouse bones. Leupeptin and antipain do not affect collagenase production and activity or the enhanced secretion of beta-glucuronidase induced by the bone-resorbing agents. They might thus act by a direct (extracellular?) inhibition of lysosomal thiol proteinases. PMID- 6272703 TI - Involvement of superoxide anion in the reaction mechanism of haemoglobin oxidation by nitrite. AB - The sigmoidal time course of haemoglobin oxidation by nitrite, involving an initial slow reaction accompanied by a subsequent rapid reaction, was extensively explored. The initial slow reaction was much prolonged by the addition of superoxide dismutase to the reaction mixture. On the other hand, in the presence of superoxide anion generated by xanthine oxidase systems, the slow phase disappeared and the reaction changed to first-order kinetics. The oxidation of intermediate haemoglobins [defined as haemoglobin tetramer in which different chains (alpha- or beta-) are in the ferric state and in the ferrous state] such as (alpha 2+ beta 3+)2 and (alpha 3+ beta 2+)2 also proceeded in a sigmoidal manner. Similar effects of superoxide anion on these reactions were observed. Since the intermediate haemoglobins such as (alpha 2+ beta 3+)2 and (alpha 3+ beta 2+)2 were found to be produced by the oxidation of haemoglobin by nitrite, the changes in oxyhaemoglobin, intermediate haemoglobins and methaemoglobin during the reaction were followed by isoelectric-focusing electrophoresis. The amounts of (alpha 2+ beta 3+)2 were larger than those of (alpha 3+ beta 2+)2 at the initial stages of the reaction, suggesting that there is a functional difference between alpha- and beta-chains in the oxyhaemoglobin tetramer. On the basis of these results, a reaction model of the haemoglobin oxidation by nitrite was tentatively proposed. The changes in oxyhaemoglobin, intermediate haemoglobins and methaemoglobin were well fitted to the simulation curves generated from the reaction model. Details of the derivation of the equations used for kinetic analysis have been deposited as Supplement SUP 50112 (5 pages) with the British Library Lending Division, Boston Spa, Wetherby, West Yorkshire LS23 7BQ, U.K. from whom copies may be obtained on the terms indicated in Biochem. J. (1978) 169, 5. PMID- 6272705 TI - Studies on the hydrophobic properties of sphingomyelinase. AB - Crude liver lysosomal sphingomyelinase (EC 3.1.4.12) displays a heterogeneous electrofocusing profile. The majority of the enzyme resolves into two major components with acidic pI values near pH 4.6 and 4.8. Several additional minor peaks of activity are seen at more basic pH values (up to pH 8.0). In the presence of 0.1% Triton X-100 (or Cutscum), the location of sphingomyelinase is shifted by about 1 pH unit to more basic pH values. Triton X-100 also increases the apparent heterogeneity of sphingomyelinase. Removal of detergent by treatment with Bio Beads SM-2 restores the acidic pI profile. This behaviour appears to be specific, since it was not shared by six glycosidases several of which hydrolyse sphingolipids. The electrofocusing profile of 3H-labelled Triton X-100 was distinct and separate from sphingomyelinase, suggesting that only a small fraction of detergent interacted directly with the enzyme. To study this behaviour in more detail we examined the effect of detergents on elution of sphingomyelinase from sphingosylphosphocholine-Sepharose. Sphingosylphosphocholine is a competitive inhibitor of sphingomyelinase (Ki 0.5 mM). Binding of enzyme was pH-dependent. Triton X-100, Cutscum and Tween 20 eluted significant amounts of enzyme at 0.01-0.02%. Total elution was achieved with up to 0.1% detergent. These data suggest that sphingomyelinase binds to neutral detergent monomers with a high degree of affinity. In excess detergent (5 7 times the critical micellar concentration) the surface charge on the protein is changed, leading to a pI shift. This behaviour probably does not occur at the active site of the enzyme, since there is no appreciable effect on substrate hydrolysis and substrate analogues were ineffective in eluting the enzyme. PMID- 6272704 TI - Effect of bile salts on the hydrolysis of gangliosides, glycoproteins and neuraminyl-lactose by the neuraminidase of Clostridium perfringens. AB - Studies were done on the effect of bile salts on the rates of hydrolysis of the N acetylneuraminyl linkages of several sialic acid-containing compounds by the neuraminidase of Clostridium perfringens. When GM3-ganglioside, two glycolipids (glycophorin and orosomucoid) and neuraminyl-lactose were used as substrates, hydrolysis was obtained even in the absence of bile salts, but addition of this detergent, below its critical micellar concentration, increased the reaction rates; above the critical micellar concentration of the detergent rates decreased again. When a second ganglioside, GM1, was used as substrate, the requirement for bile salts was absolute; hydrolysis was not observed at all without this detergent. With increasing concentrations of bile salt and in the presence of high concentrations of enzyme, rates of hydrolysis increased, reaching maximal values at fixed ratios of bile salt to GM1-ganglioside. Physical measurements showed that mixtures of bile salt and GM1-ganglioside form mixed micelles that have a higher critical micellar concentration, a lower molecular weight and greater axial ratio than the corresponding micelles of pure GM1-ganglioside. PMID- 6272706 TI - The effects of surface and macromolecular interactions on the kinetics of inactivation of trypsin and alpha-chymotrypsin. AB - The autolysis of trypsin and alpha-chymotrypsin is accelerated in the presence of colloidal silica and glass surfaces. It is proposed that adsorption of the enzymes (favoured by electrostatic factors) results in a conformational change that renders the adsorbed enzyme more susceptible to proteolytic attack. Although the adsorbed enzymes are more susceptible to proteolysis, their activity towards low-molecular-weight substrates is not affected, indicating a relatively minor conformational change on adsorption. The rates of autolysis in solution (i.e. in ;inert' vessels) are second-order for both trypsin and alpha -chymotrypsin, with rate constants of 13.0mol(-1).dm(3).s(-1) for trypsin (in 50mm-NaCl at pH8.0 at 25 degrees C) and 10.2mol(-1).dm(3).s(-1) for alpha-chymotrypsin (in 0.1m-glycine at pH9.2 at 30 degrees C). In glass vessels or in the presence of small areas of silica surface (as colloidal silica particles), the autolysis of both trypsin and alpha-chymotrypsin can show first-order kinetics. Under these conditions, saturation of the surface occurs and the fast surface proteolytic reaction controls the overall kinetic order. However, when greater areas of silica surface are present, saturation of the surface does not occur, and, since for a considerable portion of the adsorption isotherm the amount adsorbed is approximately proportional to the concentration in solution, second-order kinetics are again observed. A number of negatively charged macromolecules have been shown similarly to increase the rate of autolysis of trypsin: thus this effect, observed initially with glass and silica surfaces, is of more general occurrence when these enzymes adsorb on or interact with negatively charged surfaces and macromolecules. These observations explain the confusion in the literature with regard to the kinetics of autolysis of alpha-chymotrypsin, where first-order, second-order and intermediate kinetics have been reported. A further effect of glass surfaces and negatively charged macromolecules is to shift the pH activity curve of trypsin to higher pH values, as a consequence of the effective decrease in pH in the ;microenvironment' of the enzyme associated with the negatively charged surface or macromolecule. PMID- 6272707 TI - Erythroglycan biosynthesis in K-562 cells. Inhibition of synthesis by tunicamycin and lack of attachment to the G-protein of vesicular-stomatitis virus. AB - K-562 cells, which express foetal erythroglycan, are shown to synthesize the lipid-linked oligosaccharide intermediates commonly found in tissues and cultured fibroblasts. The addition of tunicamycin, which blocks the formation of these intermediates and thus of asparagine-linked oligosaccharides, inhibits the synthesis of erythroglycan (Mr 7000-11 000). Vesicular-stomatitis-virus infection of K-562 cells results in the glycosylation of the G-protein with the transferrin type oligosaccharide (Mr 3000), but not with the larger erythroglycan. These results suggest that, in K-562 cells, the early stages of erythroglycan biosynthesis are the same as those of the transferrin-type oligosaccharides. However, maturation of the oligosaccharide is influenced by protein structure such that erythroglycan is only expressed on specific glycoproteins. PMID- 6272708 TI - Production of superoxide radicals by soluble hydrogenase from Alcaligenes eutrophus H16. AB - The soluble hydrogenase (hydrogen-NAD+ oxidoreductase, EC 1.12.1.2) of Alcaligenes eutrophus H16 was shown to be stabilized by oxidation with oxygen and ferricyanide as long as electron donors and reducing compounds were absent. The simultaneous presence of H2, NADH and O2 in the enzyme solution, however, caused an irreversible inactivation of hydrogenase that was dependent on the O2 concentration. The half-life periods of 4 degrees C under partial pressures of 0.1, 5, 20 and 50% O2 were 11, 5, 2.5 and 1.5 h respectively. Evidence has been obtained that hydrogenase produces superoxide free radical anions (O2-.), which were detected by their ability to oxidize hydroxylamine to nitrite. The correlation between O2 concentration, nitrite formation and inactivation rates and the stabilization of hydrogenase by addition of superoxide dismutase indicated that superoxide radicals are responsible for enzyme inactivation. During short-term activity measurements (NAD+ reduction, H2 evolution from NADH), hydrogenase activity was inhibited by O2 only very slightly. In the presence of 0.7 mM-O2 an inhibition of about 20% was observed. PMID- 6272709 TI - Structural studies of eukaryotic cytochrome c modified at methionine-65. AB - 1H n.m.r. spectra were recorded in both oxidation states for the following species: tuna cytochrome c, tuna [carboxymethylmethionine-65]cytochrome c, horse cytochrome c and horse [homoserine-65]cytochrome c. The experiments give the assignments of the singlet methyl resonances of methionine-65 and the N-terminal acetyl group. The modification at methionine-65 is shown to cause an extremely small structural perturbation to one part of the molecule close to the site of modification. PMID- 6272710 TI - Purification and properties of D-ribulokinase and D-xylulokinase from Klebsiella aerogenes. AB - The D-ribulokinase and D-xylulokinase of Klebsiella aerogenes were purified to homogeneity from Escherichia coli K12 construct strains that synthesized these enzymes constitutively. The D-ribulokinase, which is encoded in the ribitol operon, is active as a dimer of 60 000 subunit mol.wt., whereas the D xylulokinase, which is encoded in the D-arabitol operon, is active as a dimer of 54 000 subunit mol.wt. The amino acid compositions and N-terminal sequences of both pentulokinases are reported. The Kapp. values of the enzymes for their D pentulose substrates were determined, and the D-ribulokinase was shown to have a low-affinity side-specificity for ribitol and D-arabitol. These results are discussed in the context of the evolution of the Klebsiella aerogenes pentitol operons. PMID- 6272711 TI - The hydrolysis of phosphatidylinositol monolayers at an air/water interface by the calcium-ion-dependent phosphatidylinositol phosphodiesterase of pig brain. AB - 1. The activity of Ca2+-dependent phosphatidylinositol phosphodiesterase (EC 3.1.4.10) of pig brain against [32P]phosphatidylinositol monolayers at an air/water interface has been measured. As the monolayer pressure was increased a sharp cut-off of enzymic hydrolysis occurred at 33 X 10(-3) N/m. 2. The addition of either phosphatidic acid, phosphatidylglycerol or oleyl alcohol increased the film pressure at which cut off occurred, as well as increasing the rate of hydrolysis at lower pressures. 3. The rate of hydrolysis, but not the cut-off pressure, was markedly increased by oleic acid and slightly increased by phosphatidylethanolamine. 4. Phosphatidylcholine, palmitoylcholine and octadecylamine decreased the cut-off pressure, as well as the enzymic activity below this pressure. 5. Stearic acid and stearyl alcohol had no effect on either the cut-off pressure or the activity. 6. All activators decreased the length of the lag phase before enzyme activity began, and phosphatidylcholine increased it. 7. These results are compared with the stimulatory and inhibitory effects of various amphiphiles observed previously with phosphatidylinositol dispersions [Irvine, Hemington & Dawson (1979) Eur. J. Biochem. 99, 525-530], and their possible relevance to the control of the phosphatidylinositol phosphodiesterase in vivo are discussed. PMID- 6272712 TI - Dependence on freezing of the geometry and redox potential of type 1 and type 2 copper sites of Japanese-lacquer-tree (Rhus vernicifera) laccase. AB - The room-temperature e.p.r. spectrum of the Japanese-lacquer-tree (Rhus vernicifera) laccase shows A parallel (the hyperfine splitting constant) and g parallel values of both the Type 1 and Type 2 Cu appreciably different from those measured at liquid-N2 temperature. The geometry of the sites, as inferred from the room-temperature e.p.r. parameters, is more consistent with their redox properties. A rough correlation is found between A parallel and g parallel values and redox potential of the blue copper in several enzymes. PMID- 6272713 TI - A method for the preparation of low-pH dodecyl sulphate/polyacrylamide-gradient gels. AB - 1. A low-pH lithium dodecyl sulphate/polyacrylamide-gradient slab-gel system, suitable for electrophoresis, is described, and the migration properties of standard proteins are compared on this and conventional high-pH gels. 2. Cytochrome oxidase may be partially resolved into its component polypeptides. The order of migration of these is, however, dependent on the pH of the gel system. PMID- 6272714 TI - N.m.r. and e.p.r. characterization of [4-carboxy-2,6-dinitrophenyl lysine]cytochromes c. AB - Monosubstituted [4-carboxy-2,6-dinitrophenyl-lysine]cytochromes c were investigated by n.m.r. and e.p.r. Modification of Lys-13 or Lys-72 in ferricytochrome c by 4-chloro-3,5-dinitrobenzoate yields either of two different conformers that are rapidly exchanging in the native form. The equilibrium involves small local changes in the conformation of Met-80 (the sixth ligand) and Phe-82, as a result of whether Lys-13 is the 'on' or 'off' position in the Lys-13 -Glu-90 salt bridge. PMID- 6272715 TI - Neutral glycolipids and gangliosides of concanavalin A-selected SV3T3 revertant cells and of normal and SV40-transformed Balb/c 3T3 cells. AB - The structural analysis of neutral glycolipids and gangliosides of the SV40 transformed Balb/c3T3 cells (SV3T3 cells) and concanavalin A-selected SV3T3 revertant cells, both compared with untransformed Balb/c3T3 cells, has shown: (i) a content of neutral glycolipids in revertant cells near to that found in the untransformed parental cells; (ii) a similar decrease of the higher gangliosides in transformed and revertant cells; (iii) a content of ganglioside GM3 in revertant cells much higher than that found in both SV3T3 and untransformed Balb/3T3 cells. The possible role of ganglioside GM3 in growth control is discussed. PMID- 6272716 TI - A study of the magnetic properties of haem a3 in cytochrome c oxidase by using magnetic-circular-dichroism spectroscopy. AB - M.c.d. (magnetic-circular-dichroism) spectroscopy was used to study the magnetization properties of the haem centres in cytochrome c oxidase with magnetic fields of between 0 and 5.3 T over the temperature range 1.5--200 K. The oxidized, oxidized cyanide and partially reduced cyanide forms of the enzyme were studied. In the oxidized state only cytochrome a3+ is detectable by m.c.d. spectroscopy, and its magnetization characteristics show it to be a low-spin ferric haem. In the partially reduced cyanide form of the enzyme cytochrome a is in the diamagnetic low-spin ferrous form, whereas cytochrome a3--CN is e.p.r. detectable and gives an m.c.d.-magnetization curve typical of a low-spin ferric haem. In the oxidized cyanide form of the enzyme both cytochrome a and cytochrome a3--CN are detectable by m.c.d. spectroscopy, although only cytochrome a gives an e.p.r. signal. The magnetization characteristics of haem a3--CN show clearly that its ground state is an electronic doublet and that another state, probably a spin singlet, lies greater than 10 cm-1 above this. These features are well accounted for by an electronic state of spin S = 1 with a predominantly axial distortion, which leaves the doublet, Ms = +/- 1, as the ground state and the component Ms = 0 as the excited state. This state would not give an e.p.r. signal. Such an electronic state could arise either from a ferromagnetic coupling between haem a3+(3)-CN and the cupric ion, Cua3, or form a haem in the Fe(IV) state. PMID- 6272717 TI - Characterization of the partially reduced cyanide-inhibited derivative of cytochrome c oxidase by optical, electron-paramagnetic-resonance and magnetic circular-dichroism spectroscopy. AB - Optical. e.p.r. and near-infrared low-temperature m.c.d. (magnetic-circular dichroism) spectroscopy were used to characterize the partially reduced cyanide inhibited derivative of cytochrome c oxidase produced by anaerobic reductive titration with dithionite. The reductions of cytochrome a3+ and Cu2+a were followed by observation of the e.p.r. signals at g = 3.03, 2.21 and 1.5 and at g = 2.18, 2.03 and 1.99. As reduction proceeds new e.p.r. signals (g = 3.58 and 1.56) appear that quantify to give one haem per enzyme unit when a small excess of dithionite has been titrated in. The e.p.r. signal of the Cu2+a titrates in parallel with the disappearance of the band and 820nm in the optical absorption spectrum. The near-infrared m.c.d. spectrum shows the presence of the low-spin ferric haem, a3+, in the oxidized state of the enzyme, as a well-resolved positive peak at 1650nm. As reduction proceeds this band is replaced by one at 1550nm due to haem a3+(3)--CN in the partially reduced state. Hence as haem a3+(3)--CN becomes e.p.r.-detectable it also shows a near-infrared m.c.d. spectrum characteristic of a low-spin ferric haem. It is concluded that the partially reduced state of cyanide-inhibited cytochrome c oxidase contains a2+ . Cu+a . a3+(3)--CN . Cu+a3. PMID- 6272718 TI - Photolabelling of membrane proteins with photoactive phospholipids. AB - Photoactive probes have been introduced recently to study the hydrophobic sector of integral membrane proteins. A simple procedure to synthesize a new series of highly radioactive aryl azido-phospholipids is presented. They effectively exchange with the boundary lipids and, on illumination, the cross-link to several membrane proteins with high efficiency. The procedure and analysis of labelling of ATPase from sarcoplasmic reticulum is reported as an example. The advantages in using these photoactive phospholipids are discussed together with some information obtained on their use. PMID- 6272719 TI - Evidence that the lack of high catalytic activity of thiolsubtilisin towards specific substrates may be due to an inappropriately located proton-distribution system. Demonstration of highly nucleophilic character of the thiol group of thiolsubtilisin in the catalytically relevant ionization state of the active centre by use of a two-protonic-state reactivity probe. AB - The active centre of the semi-synthetic enzyme thiolsubtilisin was investigated by studying the kinetics of the reaction of the thiol group of cysteine-221 with the thiol-specific two-protonic-state reactivity probe 2,2'-dipyridyl disulphide. The three-states criterion [Brocklehurst (1974) Tetrahedron 30, 2397-2407] was used to provide definitive evidence of the existence of a thiol--imidazole interactive system in acidic media in which the sulphur atom possesses highly nucleophilic character. The lack of catalytic competence of thiolsubtilisin despite its possession of the requisite nucleophilic capability is discussed. The exceedingly high rate of reaction of thiolsubtilisin with 2,2'-dipyridyl disulphide at pH 4--5 is shown to constitute a rapid and convenient active-site titration in which intact thiol--imidazole interaction is detected even in the presence of other thiols. PMID- 6272720 TI - Cyclic AMP-independent casein/glycogen synthase kinases from pig polymorphonuclear leucocytes. AB - 1. Two cyclic AMP-independent casein/glycogen synthase kinases were purified from pig polymorphonuclear leucocytes by chromatography on phosphocellulose followed by affinity chromatography on casein-Sepharose 4B or gel filtration on Bio-Gel A 1.5m. When the affinity step was used, the specific activities were 86 and 43units/mg of protein for casein kinase 1 and 2, respectively, whereas these values were 94 and 90units/mg of protein when the gel-filtration step was used. 2. These kinases differ as follows: (a) the molecular weight of casein kinase 1 (38000) is very much lower than that of casein kinase 2 (185000); (b) the K(m) for casein (0.46mg/ml) and K(a) for Mg(2+) (0.3mm) of casein kinase 1 are lower than those of casein kinase 2 (0.90mg/ml and 1.7mm respectively); (c) KCl stimulates the phosphorylation of casein by casein kinase 1, whereas it inhibits phosvitin phosphorylation by this enzyme; on the contrary, the effect of KCl on casein kinase 2 is very similar with either casein or phosvitin as substrate; (d) although both kinases phosphorylate rabbit muscle glycogen synthase I, the ratio of glycogen synthase to casein phosphorylation by casein kinase 1 is about 4-fold greater than that by casein kinase 2. Furthermore, (32)P incorporation into glycogen synthase promoted by casein kinase 1 (3.6mol of (32)P/mol of 85000 dalton subunit) is twice that observed with casein kinase 2 (1.8mol of (32)P/mol of 85000-dalton subunit). Such a phosphorylation results in a decrease in the glucose 6-phosphate-independence ratio of glycogen synthase to 10-15 with casein kinase 1 and to 35-45 with casein kinase 2. 3. The activity of both kinases is neither stimulated by cyclic AMP, Ca(2+) and calmodulin nor inhibited by cyclic AMP-dependent protein kinase inhibitor protein. 4. No phosphorylation kinase activity was observed with casein kinase 1 and 2 at either pH6.8 or 8.2 in the presence of Ca(2+). 5. Activities of both kinases on casein and glycogen synthase decreased in parallel when incubated at 50 degrees C. PMID- 6272721 TI - Isolation and characterization of calmodulin from an insulin-secreting tumour. AB - A major protein constituent of a rat islet cell tumour that exhibited Ca2+ dependent changes in electrophoretic mobility has been purified to homogeneity and compared in its physicochemical and biological properties with bovine brain and rat brain calmodulin (synonymous with phosphodiesterase activator protein, calcium-dependent regulator, troponin C-like protein and modulator protein). The protein, like these calmodulins, contained trimethyl-lysine, exhibited a blocked N-terminus and had an identical amino-acid composition and molecular weight on sodium dodecyl sulphate/polyacrylamide-gel electrophoresis. Peptide "maps' prepared after digestion of the three proteins with trypsin, papain or Staphylococcus V-8 proteinase were virtually superimposable. Ca2+ altered the electrophoretic mobilities the enhanced the native protein fluorescence in an equivalent manner with all three proteins. Equilibrium dialysis experiments demonstrated in each case the binding of 4g-atoms of calcium/mol of protein; the binding sites were equivalent and showed Kd 0.8 microM. Tumour and brain proteins were equipotent as Ca2+-dependent activators of partially purified rat brain cyclic nucleotide phosphodiesterase, and in this action were inhibited in an identical manner by trifluoperazine. The proteins also exhibited the common property of Ca2+-dependent binding to troponin I, histone H2B and myelin basic protein. The estimated tumour content of calmodulin was 450 mg/kg fresh wt., a value similar to that reported in islets of Langerhans. These results further document the validity of the islet cell tumour as an experimental model of Ca2+ mediated molecular events associated with insulin secretion. They also suggest that brain calmodulin may be substituted for endogenous calmodulin in experimental investigations into the mechanism of insulin secretion. PMID- 6272722 TI - Ca2+-dependent and Ca2+-independent degradation of phosphatidylinositol in rabbit vas deferens. AB - The effects of Ca2+ and acetylcholine on the degradation and synthesis of phosphatidylinositol in rabbit vas deferens was studied in vitro by a pulse- chase technique and by measuring the content of the phospholipid in the tissue. Ca2+-dependent degradation of phosphatidylinositol was found in slices and homogenates prelabelled with myo-[2-3H]inositol. The phosphatidylinositol content of the slices also decreased by a Ca2+-dependent mechanism. On the other hand, removal of intracellular Ca2+ with the ionophore A23187 and EGTA increased the amount of phosphatidylinositol. These results indicate that the intracellular Ca2+ concentration has an important role in regulating the phosphatidylinositol content of the tissue. Increasing the extracellular K+ concentration, which causes an increase in plasma-membrane Ca2+ permeability, did not enhance phosphatidylinositol breakdown nor decrease its tissue content. However, phosphatidylinositol synthesis was clearly inhibited. After stimulation of the smooth muscle with acetylcholine, degradation of phosphatidylinositol was enhanced. Furthermore, the content of phosphatidylinositol in the tissue also decreased. These phenomena were evident even in the absence of Ca2+. The acetylcholine-induced degradation of phosphatidylinositol was blocked by the muscarinic antagonist atropine, but not by the nicotinic antagonist (+) tubocurarine. The acetylcholine-induced decrease in the phosphatidylinositol content of the tissue led to the compensatory synthesis of phosphatidylinositol. Synthesis was separated from degradation in the same tissue. Compensatory synthesis was inhibited by acetylcholine. The degradation of phosphatidylinositol induced by acetylcholine was not inhibited by 8-bromoguanosine 3':5'-cyclic monophosphate, indicating that the degradative process was not mediated by an increase in the cyclic nucleotide. PMID- 6272723 TI - Stimulation of inorganic-phosphate incorporation into phosphatidylinositol in rat thoracic aorta mediated through V1-vasopressin receptors. AB - Vasopressin stimulates the incorporation of [32P]Pi into phosphatidylinositol but not into other phospholipids in rat thoracic aorta strips. The relative abilities of three vasopressin analogues to stimulate phosphatidylinositol labelling in rat aorta are similar to their relative pressor potencies in vivo and to their relative potencies in stimulating the metabolism of rat hepatocytes, but very different from their relative antidiuretic potencies. The vasopressor antagonist [1-(beta-mercapto-beta, beta-cyclopentamethylenepropionic acid),8 arginine]vasopressin competitively inhibits [Arg8]vasopressin-stimulated phosphatidylinositol labelling in rat aorta with a pA2 of 8.1. It is concluded that the Ca2+-mobilizing vasopressin receptors (V1-receptors) of the rat aorta stimulate phosphatidylinositol metabolism, probably by enhancing phosphatidylinositol breakdown. PMID- 6272724 TI - Receptor-mediated gonadotropin action in the ovary. Action of cytoskeletal element-disrupting agents on gonadotropin-induced steroidogenesis in rat luteal cells. AB - The role of the cellular cytoskeletal system of microtubules and microfilaments on gonadotropin-stimulated progesterone production by isolated rat luteal cells has been investigated. Exposure of luteal cells to human choriogonadotropin resulted in a stimulation of cyclic AMP (4-7-fold) and progesterone (3-4-fold) responses.l Incubation of cells with the microfilament modifier cytochalasin B inhibited the gonadotropin-induced steroidogenesis in a dose- and time-dependent manner. The effect of cytochalasin B on basal production of steroid was less pronounced. Cytochalasin B also inhibited the accumulation of progesterone in response to lutropin, cholera enterotoxin, dibutyryl cyclic AMP and 8-bromo cyclic AMP. The inhibition of steroidogenesis by cytochalasin B was not due to (a) inhibition of 125I-labelled human choriogonadotropin binding to luteal cells, (b) inhibition of gonadotropin-stimulated cyclic AMP formation or (c) a general cytotoxic effect and/or inhibition of protein biosynthesis. Cytochalasin D, like cytochalasin B, inhibited gonadotropin- and 8-bromo cyclic AMP-stimulated steroidogenesis. Although cytochalasin B also blocked the transport of 3-O-methyl glucose into luteal cells, cytochalasin D was without such an effect. Increasing glucose concentration in the medium, or using pyruvate as an alternative energy source, failed to reverse the inhibitory effect of cytochalasin B. The anti microtubular agent colchicine failed to modulate synthesis and release of progesterone by luteal cells in response to human choriogonadotropin. These studies suggest that the cellular microfilaments may be involved in the regulation of gonadotropin-induced steroidogenesis. In contrast, microtubules appear to be not directly involved in this process. PMID- 6272725 TI - Phosphorylation of nuclear and DNA-binding proteins in proliferating and quiescent mammalian cells. AB - The dependence of cell proliferation on nuclear protein phosphorylation was studied with exponential-phase and stationary-phase cultures of Chinese-hamster ovary cells. Nuclear proteins were fractionated, according to their DNA-binding affinities, by using sequential extractions of isolated nuclei with increasing concentrations of NaCl. When viable whole cells were labelled with H332PO4, phosphorylation of nuclear proteins was found to be lower in quiescent cells than in proliferating cells. Phosphorylation of nuclear proteins soluble in 0.30M-NaCl (less than 50% of these proteins bind to DNA) was greater than for those proteins soluble in higher salt concentrations (80-100% of these proteins bind to DNA). Cyclic AMP enhanced the phosphorylation of nuclear proteins soluble in 0.3 m-NaCl by 40-50%, and this stimulation was independent of cell growth. Cyclic AMP also increased the phosphorylation of nuclear proteins soluble in 0.6M-NaCl and 2.0M NaCl by 40-50% in exponential-phase cultures, but not in stationary-phase cultures. Several examples of specific phosphorylation in response to cyclic AMP were observed, including a 35000-mol.wt. protein in the 0.30 M-NaCl-soluble fraction and several proteins larger than 100000 molecular weight within this fraction. A major peptide of molecular weight approx. 31000 extracted with 0.6M NaCl was also phosphorylated. Its phosphorylation was independent of cyclic AMP in exponential-phase cultures, and it was not phosphorylated in plateau-phase cells. These changes in cell-growth-dependent phosphorylation occurred in the absence of any apparent qualitative changes in the nuclear protein molecular weight distributions. These data demonstrate that (1) phosphorylation of nuclear proteins is dependent on the culture's proliferative status, (2) both cyclic AMP dependent and cyclic AMP-independent specific phosphorylation occurs, and (3) the cyclic AMP-dependent growth-independent phosphorylation that occurs does not appear to be a modification of DNA-binding proteins, whereas the cyclic AMP dependent growth-dependent phosphorylation does involve modification of DNA binding proteins. PMID- 6272726 TI - A comparison of ornithine decarboxylases from normal and SV40-transformed 3T3 mouse fibroblasts. AB - Ornithine decarboxylase (L-ornithine carboxy-lyase, EC 4.1.1.17) has been purified from 3T3- and SV40-transformed 3T3 mouse fibroblasts by affinity chromatography, and the physicochemical properties of the two enzymes compared. Measured properties include molecular weight of the active species, subunit molecular weight and specific activity of the purified enzymes, kinetic parameters, thermostability, degradation rate in vivo and immunological cross reactivity. Although crude extracts of the transformant possess more ornithine decarboxylase activity per mg of protein than the parent strain, there is no evidence for the appearance of an altered form of the enzyme in these cells. The results reported in the present paper indicate that the increased ornithine decarboxylase activity in the transformed cells is the result of higher enzyme biosynthesis de novo. PMID- 6272727 TI - Corticotropin-(1--24)-tetracosapeptide affects protein phosphorylation and polyphosphoinositide metabolism in rat brain. AB - 1. Effects of corticotropin-(1--24)-tetracosapeptide on the endogenous phosphorylation of proteins and lipids were studied in a membrane/cytosol fraction prepared from a lysed crude mitochondrial/synaptosomal fraction. 2. The labelling of proteins and lipids was monitored by incubation of the subcellular fraction for 10s with [gamma-32P]ATP. 3. The phosphorylation of proteins was dose dependently inhibited by the peptide (40% of control incubations at 100 microM corticotropin). 4. Of the membrane phospholipids only phosphatidylinositol phosphate, phosphatidylinositol bisphosphate and phosphatidic acid became labelled. Corticotropin dose-dependently increased the formation of phosphatidylinositol bisphosphate and inhibited the production of phosphatidic acid (470% and 50% respectively of control incubations, at 100 microM of the peptide) and had no effect on phosphatidylinositol phosphate. 5. Phosphatase activity was observed to act on phosphatidylinositol bisphosphate, phosphatidylinositol phosphate and phosphoprotein but not on phosphatidic acid. 6. Corticotropin interacted with the kinases rather than with the phosphatases. 7. The formation of phosphatidylinositol bisphosphate and phosphatidic acid was maximal at 1--10mM-Mg2+ in the absence of Ca2+, and the production of phosphatidylinositol phosphate was maximal at 30mM-Mg2+. 8. The basal value of lipid phosphorylation decreased with increasing Ca2+ concentration. 9. Ca2+ abolished the effect of corticotropin on phosphatidylinositol bisphosphate formation (470%, 190% and 100% of control incubations at respectively 0, 0.1 and 1 mM-Ca2+). 10. The data provide evidence that the effects of corticotropin on protein phosphorylation and on polyphosphoinositide metabolism in brain membranes are related. PMID- 6272728 TI - The effects of N-alkylmaleimides on the activity of rat liver glucose 6 phosphatase. AB - A series of N-alkylmaleimides has been synthesized and used to investigate the thiol groups that are essential for the activity of rat liver microsomal glucose 6-phosphatase. All of the N-alkylmaleimides inactivated glucose 6-phosphatase when preincubated with microsomes (microsomal fractions) at pH 6.5 and 30 degrees C. When enzyme activity was assayed in intact microsomes, the inactivation was non-linear with respect time, showing an initial rapid phase followed by a slower secondary phase. During the initial rapid phase the inactivation may apparently be completely reversed by disrupting the microsomal membrane with detergent. However, after longer exposure to N-alkylmaleimides the reversal is no longer complete. This observation was explained by the results obtained from studying the inactivation in detergent-disrupted microsomes. In this case glucose 6 phosphatase was also completely inactivated, but much more slowly than was seen in intact microsomes, and the process was linear with respect to time. When assayed in both intact and detergent-disrupted microsomes, glucose 6-phosphatase inactivation was dependent on the number of carbon atoms in the alkyl side chain of the N-alkylmaleimides; this dependence was much more marked in disrupted microsomes. Analysis of the data showed that in neither case was there a saturating effect at high concentrations of maleimide. The data have been interpreted to suggest that there are are least two thiol groups essential for activity located in two separate non-polar regions of the membrane-enzyme system. The conclusions are discussed in the light of the current model for the microsomal glucose 6-phosphatase system. PMID- 6272729 TI - Characterization of human somatotropin binding to detergent-solubilized lactogenic receptors from rat liver. AB - Lactogenic receptors from rat liver microsomal fraction ('microsomes') were extracted by treatment with 1% (w/v) Triton X-100. Triton X-100 exerts an inhibitory effect on both the binding reaction and the separation of the free hormone from the complex. The association and dissociation of 125I-labelled human somatotropin are time- and temperature-dependent processes. The association rate constant, k1, is 6.7 x 10(6) mol . litre-1 . min-1 at 25 decrees C, and the dissociation rate constant, k-1, is 1.1 x 10(-3) min-1 at 25 degrees C. Scatchard analysis of saturation data reveals the existence of a single class of receptors and that solubilization leads to a slight decrease in affinity and a sharp increase in binding capacity. The dissociation constant, Kd, of the solubilized preparation is 0.22 nM and the binding capacity 2900 fmol/mg of protein. Similar results were obtained from competition experiments. Binding of 125I-labelled human somatotropin to the solubilized receptors is specifically inhibited by hormones with lactogenic activity. Incubation of the solubilized preparation with trypsin resulted in an 80% decrease in binding activity. The solubilized form of the receptor has a slightly increased sensitivity to the inactivation by trypsin, heat and extremes of pH, with respect to the membrane-bound form. PMID- 6272730 TI - Active transport of charged substrates by a proton/sugar co-transport system. Amino-sugar uptake in the yeast Rhodotorula gracilis. AB - 1. In the yeast Rhodotorula gracilis several amino sugars were actively transported. Glucosamine, which is largely protonated at physiological pH (pK 7.75) was used as a model substrate. At pH 6.75 its half-saturation constant was 1 mM and the maximal velocity was 50 nmol/min per mg dry wt. 2. Amino sugars were taken up via the monosaccharide carrier. The transport of glucosamine was strongly restricted by monosaccharides. D-Xylose inhibited competitively the uptake of glucosamine. The inhibition constant was 1 mM. Cells preloaded with D xylose showed exchange transport on subsequent addition of glucosamine. 3. Transport of glucosamine was energized by the membrane potential. Uncoupling agents such as carbonyl cyanide m-chlorophenyl-hydrazone and the lipophilic cation TPP+ (tetraphenylphosphonium ion) at concentrations that depolarized the membrane potential inhibited the uptake of glucosamine. Conversely the transport of glucosamine partly dissipated the membrane potential, which was monitored by radioactively labelled lipophilic cations. 4. The translocated charges were electrically compensated by the extrusion of protons and K+ (1 glucosamine molecule/0.85 H+ + 0.15 K+). 5. An increase of the pH in the range 4.75-8.75 lead to a decrease of the half-saturation constant from 5 mM to 1 mM and to an optimum of the maximal velocity at pH 6.75. We suggest that this fair constancy is due to the carrier not distinguishing between the protonated form of glucosamine (pH less than 7.75) and the deprotonated form (pH greater than 7.75). The increase of V(T) (maximal transport velocity) between pH 4.75 and 6.75 is due to the increase of the membrane potential: the decrease between pH 6.75 and 8.75 is due to the deprotonization of the carrier. PMID- 6272731 TI - Non-involvement of nucleolytic activities in the early effect of dimethylnitrosamine on the content of free mRNA in mouse liver. AB - 1. After dimethylnitrosamine (DMNA) administration to mice, the content of poly(A)-containing RNA decreases rapidly in the postmicrosomal fraction of the liver. We report here that the loss of free mRNA is not a result of increased nucleolytic activity. On the contrary, a decreased activity of microsomal endonuclease, assayed by its effect on polyribosomal mRNA, was demonstrated already 15 min after the administration of DMNA at 37.5 mg/kg body wt. The loss of activity was more pronounced in the rough than in the smooth membranes. Total detergent-released microsomal nucleases, as assayed by use of labelled poly(U) as substrate, showed a less rapid decline. No corresponding increase in enzyme activities was observed in the postmicrosomal fraction. 2. The dimethylnitrosamine effect on the microsomal endonuclease was not accounted for by altered lysosomal contamination of the microsomal fraction. 3. No early effect of dimethylnitrosamine administration was found on the cytoplasmic ribonuclease inhibitor. PMID- 6272732 TI - The composition of intracellular granules from the metal-accumulating cells of the common garden snail (Helix aspersa). AB - Certain cells in the hepatopancreas of the common garden snail (Helix aspersa) contain intracellular granules that are sites of metal-ion accumulation. These granules have been extracted and investigated by u.v. and i.r. spectroscopy, atomic-absorption spectroscopy, X-ray microanalysis, thermogravimetric analysis, enzymic assay and microanalysis. The deposits contain about 18% (w/w) water, 5% (w/w) organic matter and 76% (w/w) inorganic material of which the main components are Ca2+, Mg2+ and P2O7(4)-. The possible origin of these granules is discussed, as is their role in detoxifying heavy-metal ions. PMID- 6272734 TI - Inactivation of oestrogen receptor in vitro by nuclear dephosphorylation. AB - 1. Nuclei of the calf uterus are endowed with an activity inactivating crude oestrogen-receptor complex. This activity has been partially purified. It shows a very high affinity for the oestrogen-receptor complex (Km = 0.8 X 10(-9) mol of specific [3H]oestradiol-17 beta-binding sites/l) as well as for the oestrogen free receptor (Km = 1.5 X 10(-9) mol of specific [3H]oestradiol-17 beta binding sites/l). 2. The nuclear receptor-inactivating activity is enhanced by dithiothreitol and inhibited by several phosphatase inhibitors as well as by 4 nitrophenyl phosphate, as well known phosphatase substrate. This inhibition shows that a dephosphorylation process is required for the receptor inactivation. 3. The purified nuclear activity also inactivates pure receptor and phosphatase inhibitors prevent this inactivation. From these observations it appears that receptor inactivation is due to a nuclear phosphatase directly acting on the oestrogen receptor. 4. The nuclear localization of the receptor-inactivating activity, its high affinity for specific oestrogen binding sites and, as previously reported, its presence only in oestrogen target tissues suggest that this activity is the same as that involved in the nuclear loss of the receptor observed in intact cells. PMID- 6272733 TI - Direct measurement of the increase in intracellular free calcium ion concentration in response to the action of complement. AB - 1. The effect of rabbit anti-(pigeon erythrocyte) antibodies plus human complement on the concentration of intracellular free Ca2+ in sealed pigeon erythrocyte 'ghosts' was investigated with the photoprotein obelin. 2. The addition of human serum, as a source of complement, to 'ghosts' coated with antibody caused a rapid increase in intracellular free Ca2+ after a lag of 20-40 s, as detected by an increase in obelin luminescence. 3. The increase in obelin luminescence could not be explained by release of obelin into the medium. It was also Ca2+-dependent in that extracellular EGTA abolished the effect and intracellular EGTA inhibited it and required the complete terminal complex (C56789). No effect was seen with C5678. 4. The concentration of intracellular free Ca2+ before addition of complement was approx. 0.3 microM. This increased to a maximum of 5-30 microM after complement addition and then remained constant for at least 1-2 min. 5. Antibody plus complement induced a rapid increase in 42K+ efflux and an inhibition of cyclic AMP formation. 6. When partially purified complement components (C5b-9) were used in 'reactive lysis' it was possible to inhibit the release of macromolecules from pigeon erythrocyte 'ghosts' by extracellular EGTA. 7. It was concluded that the increase in intracellular free Ca2+ concentration caused by anti-cell antibody plus complement occurred before cell lysis and may be involved in the mechanism of complement-induced cell injury. PMID- 6272736 TI - Phorbol esters induce synthesis of thromboplastin activity in human monocytes. AB - 12-O-Tetradecanoylphorbol 13-acetate (TPA), phorbol 12,13-diacetate and phorbol 12,13-didecanoate were all potent inducers of thromboplastin activity in human monocytes in vitro, whereas 4 alpha-phorbol 12,13-didecanoate and 4 alpha-phorbol had no such effect. A concomitant increase in titrable apoprotein III antigen was found (apoprotein III is the protein component of thromboplastin). The increase was inhibited by cycloheximide and actinomycin D and partly by alpha-amanitin. The increase of thromboplastin activity was therefore most likely due to synthesis de novo of apoprotein III. The response was approximately halved in the absence of serum or Ca2+. Retinol had a weak inhibitory effect, and retinoic acid was inhibitory only at concentrations that also induced signs of cytotoxicity. TPA caused an initial rise in monocyte cyclic AMP concentration of about 90-120 min duration. No increase in 45Ca2+ influx was induced over 2 h. Good correlation exists between induction of apoprotein III synthesis in monocytes in vitro and mouse skin-tumour promotion in vivo by the various phorbol derivatives. Substances inactive in tumour promotion do not induce the synthesis of apoprotein III. General activating and cytotoxic effects of TPA were monitored by determining release of lysozyme, beta-glucuronidase and lactate dehydrogenase. PMID- 6272735 TI - The nature of the microfibrillar glycoproteins of elastic fibres. A biosynthetic study. AB - 1. Cell cultures propagated from foetal bovine ligamentum nuchae synthesized and secreted two glycoproteins, designated MFP I and MFP II, that are closely related to elastic-fibre microfibrils. Glycoproteins MFP I (apparent mol.wt. 150 000) and MFP II (apparent mol.wt. 300 000) were metabolically labelled, separated from other culture-medium components by immunoprecipitation with a specific anti (microfibrillar protein) serum, and analysed by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis and sodium dodecyl sulphate/gel filtration chromatography. 2. Ligament cells also synthesized and secreted fibronectin, but salt-fractionation and immunoprecipitation studies with a specific anti-(cold-insoluble globulin) serum established that neither glycoprotein MFP I nor glycoprotein MFP II was related to fibronectin. 3. The secretion of glycoprotein MFP I, but not that of glycoprotein MFP II, was enhanced by the addition of ascorbate to the culture medium. 4. Ascorbate supplemented ligament cells incorporated [3H]proline into glycoprotein MFP I, and 36% of the nondiffusible proline residues were hydroxylated, exclusively as 4 hydroxy[3H]proline. Less than 1% of the total proline residues in [3H]proline labelled glycoprotein MFP II were hydroxylated. 5. Ascorbate-supplemented cells incorporated [14C]lysine into glycoprotein MFP I and 30% of the non-diffusible lysine residues were hydroxylated. 6. Newly secreted glycoprotein MFP I was digested by highly purified bacterial collagenase to yield polypeptide fragments of apparent mol.wts. 50 000 and 30 000. Glycoprotein MFP II was not digested by bacterial collagenase. 7. The results suggest that elastic-fibre microfibrils are composed of a novel collagenous glycoprotein MFP I in association, as yet undefined, with a non-collagenous glycoprotein MFP II. PMID- 6272737 TI - Reactivity of photoreduced cytochrome aa3 complexes with molecular oxygen. AB - Cytochrome c oxidase (ox heart cytochrome aa3) is reduced on illumination in the presence of a photocatalyst system containing deazaflavin and EDTA. The photo reduced enzyme reacts with oxygen at neutral pH to give a form of ferric enzyme, whereas a corresponding sample partially reduced by light in the absence of any photocatalyst reacts with oxygen to give an oxyferri species ('oxygenated' enzyme). Reduction by the photocatalyst system at an alkaline pH value (9.0) also gives rise to fully reduced oxidase (both haem groups ferrous). At these pH values the immediate product after oxygen addition is a species with a 605-606 nm absorption band, not identical with ferrous cytochrome a, but capable of oxidizing added cytochrome c. This intermediate, which is unstable at neutral pH, may be analogous to the 'compound B' obtained by Chance and co-workers [Chance, Saronio & Leigh (1975) J. Biol. Chem. 250, 9226-9237; Chance, Saronio & Leigh (1979) Biochem. J. 177, 931-941] at low temperatures. PMID- 6272738 TI - Phospholipid requirements for the reconstitution of complex-III vesicles exhibiting controlled electron transport. AB - Phospholipid requirements for the reconstitution of Complex-III vesicles exhibiting respiratory control (electron-transport control) were studied. Vesicles prepared from pure phosphatidylethanolamine gave maximal control ratios. Phosphatidylcholine alone did not support respiratory control, although these vesicles were capable of maintaining stable K+-diffusion gradients. Apparently Complex III cannot insert into a bilayer of phosphatidylcholine. Formation of mixed phosphatidylcholine/phosphatidylethanolamine (6:1, w/w) vesicles was sufficient, however, to allow Complex-III insertion and to restore respiratory control. Mixtures of acidic phospholipids with either phosphatidylethanolamine or phosphatidylcholine did not improve respiratory control over that obtained with pure phosphatidylethanolamine. Phosphatidylethanolamine from bovine heart mitochondria, soya beans or Escherichia coli was equally effective in reconstituting respiratory control, suggesting that the specificity is referable to the head group and not to the fatty-acid moiety. PMID- 6272739 TI - Respiration-linked proton translocation in the obligate methylotroph Methylophilus methylotrophus. AB - The stoicheiometries of respiration-linked proton translocation in Methylophilus methylotrophus were determined by using both the oxygen-pulse and initial-rate methods. The latter has also been used to determine leads to charge/O quotients (measured as yield K+/O quotients) in order to ascertain whether the leads to H+/O quotients might be underestimated by H+/anion symport. The results suggest that 6H+/O are translocated during NADH oxidation, and that 2H+/O are translocated during the oxidation of methanol to formaldehyde. There was no evidence for underestimation of the leads to H+/O quotients due to H+/anion symport, except by the movement of formic acid during formate oxidation. By comparing these results with the known growth efficiencies of this organism, an leads to H+/ATP quotient of close to 2 g-ions of H+/mol of ATP can be calculated. It is proposed that the respiratory chain in Methylophilus methylotrophus is arranged such that there are three sites of energy conservation for NADH oxidation, each translocating 2H+ and each linked to the synthesis of one molecule of ATP. Only the third site of energy conservation is involved in methanol oxidation. PMID- 6272741 TI - Enzymic characteristics of an ecto-cyclic AMP-dependent protein kinase in rat epididymal spermatozoa. AB - Intact spermatozoa from rat cauda epididymides possess an ecto-(cyclic AMP dependent protein kinase) activity that causes the transfer of the terminal phosphate group of ATP to the serine residues of all the histone fractions. The enzyme showed a high degree of substrate specificity for the phosphorylation of histones rather than protamine, casein and phosvitin. The cell-external-surface protein kinase requires Mg2+ for activity, and other bivalent cations such as Mn2+ and Co2+ can substitute partially for Mg2+, whereas Ca2+ and Zn2+ are potent inhibitors of the enzyme. The enzyme has markedly higher affinity for cyclic AMP than for other cyclic nucleotides for its activation, with an apparent Km value for cyclic AmP of 80 nM. Spermatozoal ecto-kinase activity is not due to contamination of broken cells or any possible cell damage during incubation and isolation of spermatozoa. There was no loss of kinase activity from the cells when washed with 2 mM-EDTA, and the histones phosphorylated by intact spermatozoa were located outside the cells. Protein kinase activity of intact cells was strongly inhibited (approx. 90%) by p-chloromercuribenzenesulphonic acid (10 microM), which is believed not to enter the cells. These data provide further support for the localization of a protein kinase on the external surface of spermatozoa. PMID- 6272742 TI - Organization of membrane lipids and proteins in human En(a-) erythrocytes that lack the major sialoglycoprotein, glycophorin A. A spin-label study. AB - Membrane fluidity was studied by electron-spin-resonance techniques in human En(a ) erythrocytes that lack the major membrane sialoglycoprotein, glycophorin A. By using stearic acid spin labels with a doxyl group in the C-12 or C-15 positions, we demonstrated that the hydrophobic core in these cells was more fluid than in normal cells. Surface-located regions in isolated En(a-) membranes, when probed with stearic acid labelled in the C-5 position, appeared more stable than in normal membranes. In isolated En(a-) membranes, protein motion was decreased when probed with a nitroxide derivative of maleimide. After incubation with anti (glycophorin A) antibodies protein motion and membrane fluidity were increased in normal membranes. This effect was observed also after spectrin depletion, which by itself increased protein motion but decreased membrane fluidity in the hydrophobic core of the membrane. The results show that membrane proteins influence the fluidity of membrane lipids. PMID- 6272743 TI - Chemical, immunological and catalytic properties of 2':3'-cyclic nucleotide 3' phosphodiesterase purified from brain white matter. AB - The amino acid composition, isoelectric point, specificity of the antibody raised and various catalytic properties were determined for 2':3'-cyclic nucleotide 3' phosphodiesterase (EC 3.1.4.37) purified from bovine brain white matter by a procedure involving solubilization with elastase (EC 3.4.21.11). PMID- 6272740 TI - Activation and attenuation of adenylate cyclase. The role of GTP-binding proteins as macromolecular messengers in receptor--cyclase coupling. PMID- 6272744 TI - Purification of rabbit bone inhibitor of collagenase. AB - 1. Rabbit bones in tissue culture synthesize an inhibitor of collagenase during the first 4 days of culture. 2. The inhibitor was purified by a combination of gel filtration, concanavalin A--Sepharose chromatography, ion-exchange chromatography and zinc-chelate affinity chromatography. 3. The purified inhibitor migrated as a single band on sodium dodecyl sulphate/polyacrylamide-gel electrophoresis and had a mol.wt. of 28000. 4. The inhibitor blocked the activity of the metalloproteinases collagenase, gelatinase, neutral proteinase III (proteoglycanase), human leucocyte collagenase and gelatinase, but not thermolysin or bacterial collagenase. The serine proteinases plasmin and trypsin were not inhibited. 5. The inhibitor interacted with purified rabbit bone collagenase with 1:1 stoichiometry. 6. The inhibitory activity was lost after incubation for 1 h at 90 degrees C, after treatment with trypsin (250 micrograms/ml) at 37 degrees C for 30 min and after reduction and alkylation. PMID- 6272745 TI - An inhibitor of collagenase from human amniotic fluid. Purification, characterization and action on metalloproteinases. AB - 1. An inhibitor of collagenase of apparent mol.wt. 28000 was isolated from term human amniotic fluid. 2. It is active against mammalian collagenases from a number of species and tissues as well as other mammalian metalloproteinases, but has no activity against bacterial metalloproteinases. 3. Activity is destroyed by treatment with either trypsin or 4-aminophenylmercuric acetate, by heat, and by reduction and carboxymethylation. 4. All the properties observed suggest that it is similar to the synthesized tissue inhibitor of metalloproteinases. PMID- 6272746 TI - Calmodulin-free skeletal-muscle troponin C prepared in the absence of urea. AB - A method is described for the rapid preparation of electrophoretically pure troponin C from rabbit skeletal-muscle myofibrils that avoids the use of urea. The three-step procedure includes extraction od the myofibrils with EDTA containing buffers, one-step elution from DEAE-Sephadex and Sephadex G-100 chromatography in the presence of EDTA. The procedure gives yields comparable with those of currently used methods that involve dissociation of the troponin complex with urea. Except for the thiol-group reactivity, troponin C produced by our method is physicochemically and functionally indistinguishable from that obtained by the classical procedure. Purified troponin C always contains traces of calmodulin. However, this contamination can be decreased to less than 0.02% by means of a second Sephadex G-100 chromatography step in the presence of EDTA. PMID- 6272747 TI - Zinc metalloenzyme properties of active and latent collagenase from rabbit bone. AB - 1. Inhibition of collagenase from rabbit bone cultures by the chelating agents 1,10-phenanthroline and EDTA is almost completely reversed by Zn2+; other metal cations are less effective in reversing the inhibition. Optimal restoration of activity is achieved at Zn2+ concentrations below that of the chelator, but excess of Zn2+ is inhibitory. 2. Prolonged incubation of collagenase with either chelator causes irreversible inactivation. This inactivation is prevented by Zn2+ at the same concentrations needed to reverse the primary inhibition. 3. Collagenase incorporates 65Zn by exchange when incubated with 1,10-phenanthroline and Zn2+ containing this radioactive isotope. The 65Zn2+ can be removed from its binding site in collagenase by 1,10-phenanthroline or EDTA. Irreversible inactivation of collagenase by chelators destroys its ability to incorporate 65Zn2+. 4. Latent collagenase, the inhibited form in which collagenase first appears in culture, behaves similarly to the active enzyme in 65Zn2+-exchange experiments, but is resistant to irreversible inactivation by chelators. 5. It is concluded that collagenase is a zinc metalloenzyme that forms an inactive and unstable apoenzyme on treatment with chelators. The bound inhibitor component of latent collagenase evidently stabilizes the apoenzyme. PMID- 6272748 TI - Dephosphorylation of pig heart pyruvate dehydrogenase phosphate complexes by pig heart pyruvate dehydrogenase phosphate phosphatase. AB - 1. Pig heart pyruvate dehydrogenase phosphate complex in which all three sites of phosphorylation were completely phosphorylated was re-activated at a slower rate by phosphatase than complex predominantly phosphorylated in site 1. The ratio of initial rates of re-activation was approx. 1:5 with a comparatively crude preparation of phosphatase and with phosphatase purified by gel filtration and ion-exchange chromatography. 2. The ratio of apparent first-order rate constants during dephosphorylation of fully phosphorylated complex averaged 1/3.8/1.3 for site 1/site 2/site 3. Only site-1 dephosphorylation was linearly correlated with re-activation of the complex throughout dephosphorylation. Dephosphorylation of site 3 was linearly correlated with re-activation after an initial burst of dephosphorylation. 3. Because dephosphorylation of site 1 was always associated with dephosphorylation of site 2, it is concluded that dephosphorylation cannot be purely random. 4. The ratio of apparent first-order rate constants for dephosphorylation of site 1 (partially/fully phosphorylated complexes) averaged 1.72. This ratio is smaller than the ratio of approx. 5 for the initial rates of re-activation. Possible mechanisms involved in the diminished rate of re activation of fully phosphorylated complex are discussed. PMID- 6272749 TI - Purification and characterization of N-acetylglutamate 5-phosphotransferase from pea (Pisum sativum) cotyledons. AB - N-Acetylglutamate 5-phosphotransferase (acetylglutamate kinase, EC 2.7.2.8) has been isolated from pea (Pisum sativum) cotyledons and purified 312-fold by using heat treatment, (NH4)2SO4 fractionation, affinity chromatography on ATP- Sepharose and ion-exchange chromatography on DEAE-cellulose. This preparation was shown on polyacrylamide-gel electrophoresis to yield one band staining with Coomassie Blue. The enzyme was shown by a variety of techniques to be composed of two different kinds of subunits, of mol.wts. 43000 and 53000 respectively. These subunits are arranged to give either a dimeric or tetrameric enzyme composed of equal numbers of each type of subunit. The dimeric and tetrameric enzyme forms are thought to be interconvertible, the equilibrium between these forms being influenced by the type of ligand bound to the subunits. Kinetic studies performed on the purified enzyme, indicated a random Bi Bi type of mechanism. The enzyme displayed apparent negative co-operativity with respect to one of its substrates, N-acetylglutamate; as a result, two Km values were found for this substrate, one at 1.9 X 10(-3) M and the other at 6.2 X 10(-3) M. A single Km value for ATP was found to be 1.7 X 10(-3) M. Allosteric regulation by arginine was also shown. A model, based on the Koshland, Nemethy & Filmer [(1966) Biochemistry 5, 365-385] Sequential model, which adequately describes the kinetic and structural properties of N-acetylglutamate 5-phosphotransferase, is presented. PMID- 6272750 TI - Effects of high-fat diets on hepatic fatty acid oxidation in the rat. Isolation of rat liver peroxisomes by vertical-rotor centrifugation by using a self generated, iso-osmotic, Percoll gradient. AB - 1. Rat liver peroxisomal fractions were isolated in iso-osmotic Percoll gradients by using vertical-rotor centrifugation. The fractions obtained with rats given various dietary treatments were characterized. 2. The effect on peroxisomal beta oxidation of feeding 15% by wt. of dietary fat for 3 weeks was investigated. High fat diets caused induction of peroxisomal beta-oxidation, but diets rich in very long-chain mono-unsaturated fatty acids produced a more marked induction. 3. Peroxisomal beta-oxidation induced by diets rich in very-long-chain mono unsaturated fatty acids can oxidize such acids. Trans-isomers of mono-unsaturated fatty acids are oxidized at rates that are faster than, or similar to, those obtained with corresponding cis-isomers. 4. Rates of oxidation of [14-14C]erucic acid by isolated rat hepatocytes isolated from rats fed on high-fat diets increased with the time on those diets in a fashion very similar to that previously reported for peroxisomal beta-oxidation [see Neat, Thomassen & Osmundsen (1980) Biochem, J. 186, 369-371]. 5. Total liver capacities for peroxisomal beta-oxidation (expressed as acetyl groups produced per min) were estimated to range from 10 to 30% of mitochondrial capacities, depending on dietary treatment and fatty acid substrate. A role is proposed for peroxisomal beta-oxidation in relation to the metabolism of fatty acids that are poorly oxidized by mitochondrial beta-oxidation, and, in general, as regards oxidation of fatty acids during periods of sustained high hepatic influx of fatty acids. PMID- 6272751 TI - The androgenic regulation of superhelical-DNA nicking-closing enzyme in rat ventral prostate. AB - The activity of superhelical-DNA nicking-closing enzyme (NC enzyme) was measured in nuclei from rat ventral prostate by a fluorimetric assay based on the binding of ethidium bromide to supercoiled phage-PM2 DNA. The nuclear concentration of NC enzyme activity declined rapidly after castration, although this response could be prevented by daily administration of dihydrotestosterone. The low NC-enzyme activity in involuted prostates (10% of normal) was restored to normal after 8-10 days of treatment with androgen. In the regenerating prostate the time course of restoration of NC-enzyme activity was not in phase with that of DNA synthesis. Examination of nucleosome repeat lengths and the arrangement of nucleosomes along the chromatin fibre revealed no differences in the structural organization of chromatin in prostates with high or low NC-enzyme activity. Together, these results suggest that the major role of NC enzyme is related to the onset and maintenance of differentiation in the prostate and that the activity of this enzyme is not expressed through gross alterations in chromatin structure. PMID- 6272752 TI - Effect of prevention of procollagen triple-helix formation on proline 3 hydroxylation in freshly isolated chick-embryo tendon cells. AB - Inhibition of procollagen triple-helix formation by the addition of cis hydroxyproline or azetidine-2-carboxylic acid increased the synthesis of 3 hydroxy[14C]proline 1.7-1.8-fold in pulse-chase experiments with freshly isolated chick-embryo tendon cells. The amount of 3-hydroxy[14C]proline, expressed as a percentage of the total 14C radioactivity in hydroxyproline, reached 8.4%. Control experiments indicated that the two analogues had no effect on the prolyl 3-hydroxylase activity of these cells. The data suggest that the time available before triple-helix formation in part regulates the extent of the 3-hydroxylation of proline in the biosynthesis of collagen in intact cells. PMID- 6272753 TI - Choline metabolism and phosphatidylcholine biosynthesis in cultured rat hepatocytes. AB - 1. Adult rat hepatocytes were isolated by collagenase perfusion and were maintained in monolayer culture for 24h. 2. Choline metabolism and phosphatidylcholine biosynthesis were studied in these cells by performing pulse chase studies at physiological concentrations (1-40 microM) of (Me-3H)-labelled or unlabelled choline in the culture medium. 3. During the 15 min pulse incubation, choline entering the cells was rapidly phosphorylated to phosphocholine or oxidized to betaine. Low concentrations of choline in the medium decreased the relative amount of choline oxidized. 4. During the 3 h chase period, the radioactivity in the phosphocholine pool was transferred to phosphatidylcholine. Very little radioactivity was associated with CDP-choline. These results provide good evidence that the rate-limiting step for phosphatidylcholine biosynthesis in these cultured hepatocytes is the conversion of phosphocholine into CDP-choline. Similar results were obtained for all concentrations of choline in the culture medium. 5. Cellular concentrations of phosphocholine were unaffected by the concentration of choline (1-40 microM) in the medium. 6. The majority of the label associated with betaine was secreted into the culture medium during the chase incubation. 7. From the pulse-chase studies, and the cellular phosphocholine concentrations, it was possible to estimate the rate of phosphatidylcholine biosynthesis (2.2, 2.8, 3.1 and 3.7 nmol/min per g wet weight of cells cultured in 1, 5, 10 and 40 microM-choline respectively for up to 4.25 h). PMID- 6272754 TI - Stimulation by glucocorticoids of protein degradation in hepatocyte monolayers. AB - 1. Protein degradation in rat hepatocytes in stationary monolayer culture was measured as release of radioactive trichloroacetic acid-soluble material from intracellular proteins labelled with [3H]leucine. 2. Glucocorticoids, but not other steroids, stimulated protein breakdown in the hepatocyte monolayers. The effects observed were greater when the cells were preincubated with the hormones, indicating that the stimulation was not immediate. In addition, the stimulation by glucocorticoids persisted for up to 4 h after hormone removal. 3. Cycloheximide and the lysosomotropic agents leupeptin and ammonia effectively blocked glucocorticoid stimulation of protein degradation. 4. Insulin blocked dexamethasone stimulation when added at the same time as the steroid, but not when added 3 h later. 5. Stimulation of protein breakdown by dexamethasone was additive with that by glucagon or dibutyryl cyclic AMP, suggesting that its mechanism of action is different from that of the latter two agents. 6. Total activities of several lysosomal enzymes were unaffected under conditions where protein breakdown was stimulated by either glucagon or dexamethasone. 7. It is suggested that, whereas glucagon, dibutyryl cyclic AMP and insulin modulate protein breakdown in these cells via changes in autophagocytosis, the stimulation by glucocorticoids is exerted independently, perhaps by stimulating the synthesis of membrane proteins essential to the autophagic process. PMID- 6272755 TI - Human placental coated vesicles contain receptor-bound transferrin. AB - Human placental coated vesicles have been purified by a method involving sucrose density-gradient centrifugation and treatment with wheat-germ agglutinin. These preparations were free of contamination by placental microvillus fragments. Crossed immunoelectrophoresis demonstrated that the coated vesicles contained a single serum protein, which was identified as transferrin. This transferrin was only observed after the vesicles were treated with a non-ionic detergent, and its behaviour during crossed hydrophobic-interaction immunoelectrophoresis suggested that a large proportion of it was receptor-bound. No other serum proteins, including immunoglobulin G, could be detected in these preparations. Receptor bound transferrin was the only antigen common to placental coated vesicles and microvilli, implying that other plasma-membrane proteins are excluded from the region of membrane involved in coated-vesicle formation. PMID- 6272756 TI - Cyclic enkephalin analogs containing a cystine bridge. PMID- 6272757 TI - Rapid phospholipase A2 stimulation and diacylglycerol cholinephosphotransferase inhibition in baby hamster kidney cells during initiation of dengue virus infection. PMID- 6272758 TI - R 24571: a new powerful inhibitor of red blood cell Ca++-transport ATPase and of calmodulin-regulated functions. PMID- 6272759 TI - Interaction of local anesthetics with calmodulin. PMID- 6272760 TI - Amicyanin: an electron acceptor of methylamine dehydrogenase. PMID- 6272761 TI - A non-mitogenic analogue of epidermal growth factor enhances the phosphorylation of endogenous membrane proteins. PMID- 6272762 TI - Effect of vanadate on phosphoryl transfer enzymes involved in glucose metabolism. PMID- 6272763 TI - The indispensibility of a mitochondrial 15K protein for the formation of the cytochrome C1-cytochrome c complex. PMID- 6272764 TI - Diversity of rat strains and tumor lines of DNA fragments homologous to an amplified 5.8 kilobase ECO R1 fragment of Novikoff hepatoma cells. PMID- 6272765 TI - cAMP-dependent protein kinase from Mucor rouxii: physical evidence of a ternary complex holoenzyme-cAMP. PMID- 6272766 TI - Solubilization and hydrophobic immobilization assay of a cAMP binding protein from Dictyostelium discoideum plasma membranes. PMID- 6272767 TI - Dopamine increases cyclic AMP concentration in the rat spleen lymphocytes in vitro. PMID- 6272768 TI - Accessibility of some regions of chromatin of leukemic chicken cells to single strand nuclease S1. PMID- 6272769 TI - Apparent pyridine nucleotide synthesis in mitochondria: an artifact of NMN and NAD glycohydrolase activity? PMID- 6272770 TI - Evidence against the specific initiation of Okazaki fragments at the internucleosomal linkers. PMID- 6272772 TI - Glutathione-hemin complex as a cytochrome P-450 model characterization of the complex and its aromatic oxidation activities. PMID- 6272771 TI - Regulation of prolyl and lysyl hydroxylase activities in cultured human skin fibroblasts by ascorbic acid. PMID- 6272773 TI - Derivative cyclic voltabsorptometry of cytochrome c. PMID- 6272775 TI - Increased sensitivity to epinephrine stimulated lipolysis during starvation: tighter coupling of the adenylate cyclase complex. PMID- 6272774 TI - 1,25-dihydroxyvitamin D3 specifically binds to a human breast cancer cell line (T47D) and stimulates growth. PMID- 6272776 TI - Rabies virus decreases agonist binding to opiate receptors of mouse neuroblastoma rat glioma hybrid cells 108-CC-15. PMID- 6272777 TI - Metal binding peptide inhibitors of vertebrate collagenase. PMID- 6272778 TI - Cyclic GMP-dependent phosphorylation of an endogenous protein from rat heart. PMID- 6272779 TI - Desensitization of the human neutrophil degranulation response: studies with 5,12 dihydroxy-6,8,10,14- eicosatetraenoic acid. PMID- 6272780 TI - Effects of prostaglandin A2 on Na+, K+-ATPase activity in basolateral plasma membrane of rat intestine in vitro. PMID- 6272781 TI - Conformational changes expected in endogenous opioid peptides upon their interaction with acidic lipids. PMID- 6272782 TI - Spin-trapping of free radicals formed during microsomal metabolism of ethylhydrazine and acetylhydrazine. PMID- 6272783 TI - Increase of Na+/K+ ATPase activity in intact rat brain synaptosomes after their interaction with phosphatidylserine vesicles. PMID- 6272784 TI - Evidence for cytosolic benzo(a)pyrene carrier proteins which function in cytochrome P450 oxidation in rat liver. PMID- 6272785 TI - Evidence that the hepatic asialoglycoprotein receptor is internalized during endocytosis and that receptor recycling can be uncoupled from endocytosis at low temperature. PMID- 6272786 TI - Inhibition of (ADP-ribose)n biosynthesis retards DNA repair but does not inhibit DNA repair synthesis. PMID- 6272787 TI - Changes in cholesterol metabolism in cultured fibroblasts from patients with Niemann-Pick disease. PMID- 6272788 TI - Characterization of and hormonal effects on subcellular fractions from xanthophores of the goldfish Carassius auratus L. PMID- 6272789 TI - Phosphorylation of smooth muscle actin by the catalytic subunit of the cAMP dependent protein kinase. PMID- 6272790 TI - Identification of essential amino acid residues in clostridium histolyticum collagenase using chemical modification reactions. PMID- 6272791 TI - A proteolipid associated with Na,K-ATPase is not essential for ATPase activity. PMID- 6272792 TI - Fractionation of arginase-loaded erythrocyte ghosts with Percoll density gradients. PMID- 6272793 TI - Regulation of protein synthesis in rabbit reticulocyte lysates: inhibition of eIF 2 phosphoprotein phosphatase by NaF, pyrophosphate and cations. PMID- 6272794 TI - Cyclic nucleotide phosphodiesterase from a particulate fraction of rat heart. Solubilization and characterization of a single enzymatic form. PMID- 6272795 TI - High incidence of sister chromatid exchanges and chromatid interchanges in the conditions of lowered activity of poly(ADP-ribose)polymerase. PMID- 6272796 TI - Immobilization of enzymes and affinity ligands to various hydroxyl group carrying supports using highly reactive sulfonyl chlorides. PMID- 6272797 TI - ACTH immunoreactivities predominating in normal human plasma are not attributable to the human ACTH 1-39 molecule. PMID- 6272798 TI - Phosphorylation of hen progesterone receptor by cAMP dependent protein kinase. PMID- 6272799 TI - The formation of superoxide radical anions by a reaction between O2, OH- and dimethyl sulfoxide. PMID- 6272800 TI - Reactivity of chemically generated superoxide radical anion with peroxides as determined by competition kinetics. PMID- 6272801 TI - Induction of arylhydrocarbon hydroxylase and blast transformation in human blood lymphocytes. PMID- 6272803 TI - Development of lipolytic response to isoproterenol during adipose conversion of OB17 preadipocyte cells. PMID- 6272802 TI - (Na (+) + K (+)-stimulated ATPase of human kidney, normal and adenocarcinoma. Phosphorylation and inhibition by antitumor proteins. PMID- 6272804 TI - Some factors affecting the interaction between actin in leukemic L1210 cells and DNASE I. PMID- 6272806 TI - Teleocidin, a tumor promoter, is a potent platelet-aggregating agent. PMID- 6272805 TI - Uptake of Lp (a) lipoprotein by cultured fibroblasts. PMID- 6272808 TI - The missing fragment of the pro-sequence of human pro-opiomelanocortin: sequence and evidence for C-terminal amidation. PMID- 6272807 TI - Adrenocorticotropin 56. Synthesis of [Cys(Cam)25]-alpha h-ACTH-(1-26) and bis [Cys 25]-alpha h-ACTH-(1-26). Dissociation of aldosterone- and corticosterone stimulating activity. PMID- 6272809 TI - Specific binding of transforming growth factor correlates with promotion of anchorage independence in EGF receptorless mouse JB6 cells. PMID- 6272811 TI - ADP-ribosyl transferase activity of cholera toxin polypeptide A1 and the effect of limited trypsinolysis. PMID- 6272810 TI - Effect of modification of cytochrome c on its reactions with superoxide and NADPH:cytochrome P-450 reductase. PMID- 6272812 TI - EPR evidence for an acceptor functioning in photosystem II when the pheophytin acceptor is reduced. PMID- 6272814 TI - Effect of spermine on the inhibition by fatty acid on AMP deaminase reaction as a control system of the adenylate energy charge in yeast. PMID- 6272813 TI - Effect of catecholamines on ornithine decarboxylase activity in the testis of immature rat. PMID- 6272815 TI - Inhibition of the neutrophil oxidative burst and degranulation by phenothiazines. PMID- 6272816 TI - Monensin blocks endocytosis of vesicular stomatitis virus. PMID- 6272817 TI - The effect of combination therapy (steroids, immunosuppressives, and plasmapheresis) on 5 mixed cryoglobulinemia patients with renal, neurologic, and vascular involvement. AB - A prospective clinical study evaluated the effects of combined chemotherapy, steroids and plasmapheresis on 5 patients with mixed cryoglobulinemia with renal or neurologic and vasculitic manifestations. Treatment included prednisone (1mg/kg/day), chlorambucil (white blood cell count at greater than 3,000/mm3), and plasmapheresis (1 to 3 liters/week). There was healing of cutaneous ulcers (3/3), improvement in renal function (4/4), and diminution of purpura (2/2), but little improvement in peripheral neuropathy. Complications included leukopenia, perineal ulcers, and osteomyelitis. PMID- 6272818 TI - Membrane action of tricyclic drugs. Spectroscopic studies of a series of phenothiazines compared with tricyclic antidepressive substances in red cell membrane, using the spin labelling technique. AB - Previous results were extended by binding studies of drugs on red cell membrane by means of the spin labelling technique. A series of phenothiazines was investigated and compared with several tricyclic antidepressive substances. It was found that phenothiazines at concentrations below 3 mumol/l decisively increase the order parameter of spin label 618, which reports on the interface of the membrane. Thus the fluidity of the interface is decreased. Spin label 616 reports on the hydrophobic membrane interior. Within this hydrophobic phase, the order is decreased by 100 mumol to 1 mmol/l of the phenothiazines. With tricyclic antidepressive substances, a rise of order parameter of spin label 618 is observed, which, however, is less steep. At about 30 mumol/l a maximum is eventually reached. At even higher concentrations of amitriptyline, nortriptyline or imipramine with spin label 616, however, no significant change of order parameter was observed indicating that perturbation of the hydrophobic membrane interior is not sensed. These findings thus corroborate our previous results, revealing that influence of phenothiazines can be measured through the polar and apolar phases of the membrane, while the antidepressive drugs primarily act on the polar phase. PMID- 6272819 TI - [Studies on the specific-adrenolytic and unspecific-cardiodepressive action of the beta-blocker acebutolol (author's transl)]. PMID- 6272820 TI - Myocardial morphology in rabbits exposed to various gas-phase constituents of tobacco smoke--an ultrastructural study. AB - Rabbits were continuously exposed to 200 ppm carbon monoxide. Using the same criteria as applied by earlier investigators for morphological myocardial damage, no histotoxic effect on myocardial morphology could be demonstrated when electron microscopic investigations were performed blindly. Similarly, exposure to 0.5 ppm hydrogen cyanide, 0.5 ppm hydrogen cyanide + 200 ppm carbon monoxide, 0.5 ppm hydrogen cyanide + 200 ppm carbon monoxide + 5 ppm nitric oxide and to 50 ppm carbonyl sulphide for 1-7 weeks had no significant effect on myocardial ultrastructure. PMID- 6272821 TI - [Mammary carcinoid: association of mucinous and solid variants (author's transl)]. PMID- 6272822 TI - [Epidemiology of gastric cancer in Chile]. PMID- 6272823 TI - [Study of the minimum inhibiting concentration of a new cephalosporin (HR 756): comparative data with other beta-lactamic antibiotics]. AB - HR 756 is a new beta-lactamase resistant cephalosporin. The activity of HR 756 was compared in vitro to that of cefamandole, cefoxitine, cefuroxime and cephalothin. A concentration of 1 microgram/ml inhibited practically all isolates of E. coli, Salmonella, Shigella, Proteus indole + and indole --, Klebsiella, etc. A lower activity in vitro has been found regarding to Enterobacter, Serratia, Citrobacter and Pasteurella. The cefotaxime has some anti-Pseudomonas activity at concentrations of 7.5-15 micrograms/ml. For most Gram-negative organisms, HR 756 was most active. The cephalothin and the cefoxitine have resulted more active regard to Staph. aureus and Bacteroides fragilis, respectively. PMID- 6272824 TI - On the presence of cytomegalovirus in the milk of normal women; correlation between breast feeding and virus excretion in infants. PMID- 6272825 TI - [The puerperium and dysnociception. I) Behavior of serum beta-endorphins in pregnant and post-partum animals]. AB - The observation that migraine patients improve during pregnancy with return of headache upon commencement of breast feeding, led Authors to investigate the involvement of Pain Suppressor System (PSS) in pregnancy and after delivery. PSS is a complex system which utilizes neurotransmitters as beta-Endorphine-Like Immunoreactivity (beta-ELI), 5-Hydroxytryptamine (5-HT), Dopamine (DA) and GABA. The Authors report preliminary results of beta-ELI behaviour in pregnancy and in the immediate post-partum period in rats. PMID- 6272826 TI - [The puerperium and dysnociception. III) Serotonin and beta-endorphin levels in pregnant rats fed L-tryptophan]. PMID- 6272827 TI - Decreased beta-adrenoceptors in polymorphonucleates in essential hypertension. AB - Number and affinity of beta-adrenoceptors in human polymorphonucleates (PMN's) have been studied in patients with essential hypertension (n = 12) and in normal subjects of the same range of age and sex (n = 16). (-)Diidroalprenolol-H3 has been utilized as ligand. Membranes preparation was performed according to Galant (5). Number of beta-adrenoceptor is significantly decreased compared with controls (p less than 0.05), both with saturation curves and Scatchard analysis. Affinity of beta-adrenoceptors toward (-)diidroalprenolol-H3 is lightly increased but not significantly. These data could be explained on the basis of the desensitization occurring in hypertensive subjects by altered plasma catecholamines level, as in experimental in vitro systems. PMID- 6272828 TI - [Neonatal adaptive phenomenon: secretion of beta-lipotropin, and beta-endorphin in the first week of life]. AB - beta Lipotrophin and beta Endorphin plasma levels have been measured in 35 newborns subdivided in 5 groups of 12, 24, 48, 72 and 168 hours of life, 10 umbilical mixed blood samples were also evaluated. After plasma extraction (3 ml) and G-75 Sephadex column chromatography, the peptides were measured by two specific RIAs. beta LPH and beta EP levels were high in umbilical cord plasma (241.0 +/- 43.3 and 70.0 +/- 8.5 pg/ml respectively). A progressive decrement was then observed until the 72th hour of life (28.1 +/- 13.2 and 8.7 +/- 4.8 pg/ml respectively) but the elevated levels found at 24th hour demonstrate an active synthesis and release from fetal and neonatal pituitary. After a slight and transient decreased activity during the first week of life, the statistically significant increase of both beta LPH and beta EP observed at seven day indicate that at this moment pituitary function is restored. PMID- 6272829 TI - Studies on microsomal metabolizing systems for mutagenesis tests. Stability of fortified fractions. AB - Rat liver S-9 microsomal fractions were fortified with NADP+, G-6-P, and MgCl2 and stored at -80 degrees C and -20 degrees C. At various times aminopyrine demethylase and lipid peroxidation were determined and compared with that of non fortified fractions. No difference in stability was noted between fortified and not fortified fractions. The known stability pattern of non fortified fractions was confirmed. Lipid peroxidation was found blocked in the fractions stored at 80 degrees C whereas a small release of malonyl dialdehyde proceeded in the frozen state at -20 degrees C, contributing to the loss of activity at this temperature. The use of pre-fortified fractions stored at -80 degrees C is justified. PMID- 6272830 TI - A pharmacokinetic and tolerance study of Ro13-9904, a new cephalosporin antibiotic. AB - 1 Six healthy male volunteers received a total of 2500 mg of a new cephalosporin antibiotic Ro13-9904 by intramuscular injection in five divided doses at intervals of 12 h. 2 No significant systemic side-effects were observed and this was confirmed haematologically and biochemically. 3 The drug was distributed following intramuscular injection reaching a mean peak plasma concentration of 55 micrograms ml-1 (range 46-66) 1 to 2 h after the first injection. 4 Monoexponential elimination of drug was demonstrated. No significant difference was recorded in the plasma half-life after the initial dose (mean 6.7 h) and at steady state (mean 6.7 h). The half-life is long compared with other cephalosporin antibiotics. 5 On the basis of the observed half-life, steady state should be reached within 48 h. A mean peak plasma concentration of 74 micrograms ml-1 (range 65-87) was recorded at steady state. Steady state plasma concentrations of Ro13-9904 with a dose of 500 mg every 13 h may be predicted from the pharmacokinetics of a single dose. PMID- 6272831 TI - A prospective study of plasma angiotensin-converting enzyme in normotensive primigravidae and their infants. AB - Plasma angiotensin-converting enzyme (ACE) has been measured prospectively throughout pregnancy, at delivery and in the puerperium in 18 normotensive primigravidae and their infants. Plasma ACE was consistently lower during pregnancy than in comparable, non-pregnant controls, but rose progressively from about 30 weeks to term. At vaginal delivery maternal and fetal ACE levels did not differ significantly. There was a steady increase in maternal ACE activity up to 6 weeks post partum, when the levels were not significantly different from non pregnant controls. No correlation could be found between plasma ACE and plasma renin activity or concentration, or plasma AII. Plasma aldosterone increased in parallel with ACE during the last ten weeks of pregnancy. PMID- 6272832 TI - Detailed analysis of the nucleosomal organization of transcribed DNA in yeast chromatin. AB - The precise chromatin structure of actively transcribed DNA in yeast has been analyzed by electrophoretic transfer of high-resolution staphylococcal nuclease and DNase I chromatin digest DNA patterns to DBM paper and hybridization with active sequence probes. The DNA patterns of the transcribed DNA sequences resemble the DNA patterns produced by digestion of bulk yeast nucleosomes. Hence, these active sequences must be arranged in "typical" nucleosome structures. Furthermore, in details of the structure, the active sequence nucleosomes look almost exactly like the average yeast nucleosome in repeat length, in the length of DNA associated with the core particle, in the amount and type of heterogeneity found within and between the oligomeric and monomeric repeat lengths of DNA, in the occurrence of discrete spacer lengths including the characteristic five nucleotide increments (i.e., 5, 15, 25, ... base pairs), and in the length of DNA between yeast nucleosomes. Early in digestion, there are some differences: increases in peak breadths (i.e., in the distribution of spacer lengths) and some preferential release of monomer DNA. These results suggest that transcribed DNA can exist in the typical (yeast) type of nucleosome organization and thus that active chromatin regions do not necessarily require profound structural rearrangements. The slight differences noted are consistent with some slight, mainly spacer, modification in the vicinity of the transcription event itself. PMID- 6272833 TI - Production of formaldehyde during metabolism of dimethyl sulfoxide by hydroxyl radical generating systems. PMID- 6272834 TI - Effects of adrenocorticotropin and cycloheximide on adrenal diglyceride kinase. AB - We studied the effects of adrenocorticotropin (ACTH) and cycloheximide on adrenal enzymes involved in phosphatidate synthesis. Treatment of rats in vivo with ACTH induced a rapid increase in phosphatide synthesis from diglyceride and ATP in adrenal homogenates, and cycloheximide treatment prevented this increase if given before ACTH and rapidly reversed the increase if given after ACTH. The stimulatory effect of ACTH appeared to be largely due to an increase in diglyceride substrate, as kinase activity was not altered. The inhibitory effect of cycloheximide, on the other hand, appeared to be due to a decrease in diglyceride kinase activity. Neither ACTH nor cycloheximide treatment had any effect on the activity of glycerol-3'-phosphate acyltransferase or phosphatidate phosphatase. Our findings suggest that (a) ACTH increases the flow of phospholipid (and their levels) throughout the entire circular pathway, i.e., phosphatidate leads to CDP-diacylglycerol leads to inositides leads to diglycerides leads to phosphatidate, and (b) a labile protein may serve to allow entry into a recycling of diglyceride in this pathway. In addition, since cycloheximide blocked carbachol-induced increases in pancreatic and salivary glandular phosphatidate synthesis resulting from phosphatidylinositol hydrolysis and consequent diglyceride generation, the putative labile protein may have widespread importance. PMID- 6272835 TI - Some redox properties of myohemerythrin from retractor muscle of Themiste zostericola. AB - Distinct semimetmyohemerythrin species are produced by one-electron oxidation of deoxymyohemerythrin and one-electron reduction of metmyohemerythrin. The former, (semimetmyo)o, changes (greater than or equal to 90%) to the latter, (semimetmyo)R, with k = 1.0 x 10(-2) s-1, delta H = 15.1 kcal mol-1 and delta S = -17 eu. Oxidation of (semimetmyo)o by Fe(CN)6(3)- rapidly produces an unstable metmyohemerythrin form which converts to the final metmyohemerythrin with k = 4.6 x 10(-3) s-1, delta H = 16.8 kcal mol-1, and delta S = -13 eu. The two met forms react at the same rate with N3-, but the unstable form reacts very rapidly with S2O4(2-) in contrast to stable metmyohemerythrin. (Semimetmyo)R or a mixture of metmyohemerythrin and deoxymyohemerythrin equilibrate very slowly to a mixture containing all three species. The rate constants for disproportionation and comproportionation are 0.89 M-1 s-1 and 9.4 M-1 s-1, respectively. EPR spectra near liquid He temperatures and optical absorption spectra have been used to characterize and measure the rates at 25 degrees C, pH 8.2, and I = 0.15 M. The comparative behavior of octameric and monomeric protein is discussed. PMID- 6272836 TI - 1 Alpha-25,26-trihydroxyvitamin D3: an in vivo and in vitro metabolite of vitamin D3. AB - A new metabolite of vitamin D3 has been isolated from the plasma of vitamin D3 treated cows and has been generated from 25(S),26-dihydroxyvitamin D3 with homogenates of vitamin D deficient chick kidney. This metabolite has been identified as 1,25,26-trihydroxyvitamin D3 by comigration with synthetic 1,25(S),26-trihydroxyvitamin D3 in four chromatographic systems, ultraviolet spectroscopy, mass spectrometry, and high-pressure liquid chromatography and mass spectrometry of derivatives. 1,25(S),26-Trihydroxyvitamin D3 is one-tenth as effective as 1,25-dihydroxyvitamin D3 in binding to the chick intestinal cytosol 1,25-dihydroxyvitamin D receptor. Either 25(S),26-dihydroxyvitamin D3 or 1,25 dihydroxyvitamin D3 can serve as precursor for in vitro production of 1,25,26 trihydroxyvitamin D3 by chick kidney tissue. PMID- 6272837 TI - Enhancement of calcium uptake and phosphatidylinositol turnover by epidermal growth factor in A-431 cells. AB - Epidermal growth factor (EGF) stimulates the incorporation of 32Pi and [3H]inositol into phosphatidylinositol (5-10-fold) in A-431 cells. EGF also stimulates the incorporation of 32Pi into phosphatidic acid (up to 10-fold). These effects are attributed to an acceleration of the turnover of phosphatidylinositol as a consequence of the binding of EGF to its membrane receptor. The extent of the phosphatidylinositol response to EGF parallels the extent of hormone binding. The phosphatidylinositol response to EGF appears to be dependent on an influx of calcium since (a) external calcium is required for the enhancement of phosphatidylinositol turnover, (2) the accumulation of 45Ca by A 431 cells is stimulated by EGF, (3) blockage of calcium influx with LaCl3 inhibits stimulation of phosphatidylinositol turnover, and (4) calcium influx via ionophore A23187 is sufficient to stimulate phosphatidylinositol turnover. Since the binding, internalization, and degradation of 125I-labeled EGF in A-431 cells are unaffected by the omission of calcium from the medium, external calcium and phosphatidylinositol turnover are not necessary for the internalization and degradation of the EGF-receptor complex. PMID- 6272838 TI - Structural and functional similarities of delta-crystallin messenger ribonucleic acids from duck and chicken lenses. AB - delta-Crystallin of the embryonic duck lens was compared with that of the embryonic chicken lens with respect to polypeptide composition, synthesis, and messenger ribonucleic acid (mRNA) sequences. Labeling experiments with [35S]methionine revealed that the duck delta-crystallin is composed of minor amounts of polypeptides with molecular weights near 50000 (50K) and 49000 (49K) and much greater amounts of polypeptides with molecular weights near 48000 (48K) and 47000 (47K), as judged by sodium dodecyl sulfate-urea-polyacrylamide gel electrophoresis. All four sizes of polypeptides were synthesized in similar relative proportions as found in vivo in a rabbit reticulocytes lysate supplemented with delta-crystallin mRNA isolated from the embryonic duck lens. Synthesis of the 48K and 47K delta-crystallin polypeptides was differentially reduced in duck lenses cultured in the presence of ouabain. This is similar to the differential reduction of synthesis of the lower molecular weight delta crystallin peptides in embryonic chicken lenses demonstrated previously. R loops formed between duck or chicken delta-crystallin mRNA and a cloned chicken delta crystallin cDNA and heteroduplexes formed between duck or chicken delta crystallin mRNA and cloned chicken genomic DNAs containing delta-crystallin sequences showed that, except for the putative 5' leader sequence, the duck and chicken delta-crystallin mRNAs have extremely similar nucleotide sequences. These data indicate considerable conservation of delta-crystallin throughout the approximately 100 million years of divergence between ducks and chickens. The findings also suggest a possible relationship between the structure of delta crystallin mRNA and the differential reduction in synthesis of the lower molecular weight delta-crystallin polypeptides in ouabain-treated lenses of ducks and chickens. PMID- 6272840 TI - Protein influences on porphyrin structure in cytochrome c: evidence from Raman difference spectroscopy. AB - To probe the details of protein heme interactions, we have developed a Raman difference spectroscopic technique, which allows reliable detection of very small, approximately equal to 0.01 cm-1, frequency differences. When this technique is applied to heme proteins, structural differences in the protein which perturb the porphyrin macrocycle may be examined by obtaining Raman difference data on the porphyrin vibrational modes which are strongly enhanced in the Raman spectrum produced with visible laser excitation. We report here Raman difference spectroscopic data on cytochromes c from 24 species. The differences in the Raman spectrum of the porphyrin between the cytochromes c of any two species are small, confirming that all of the cytochromes we have examined have the same "cytochrome fold". However, many small (0.02-2 cm-1) but systematic differences were detected which indicate structural differences among these proteins. These differences could be classified into three different groups and interpreted in terms of different types of structural variations resulting from specific differences in the amino acid sequences. First, direct interactions between near-heme residues and the porphyrin influence the electron density in the pi orbitals of the porphyrin macrocycle. Second, variation in the residue at position 92, far removed from the heme, affects the frequency of the core-size marker line at 1584 cm-1. Third, the conformation near cysteine 14 affects the shape of the Raman mode which is sensitive to the pyrrole ring substituents (approximately 1313 cm-1). From these data we conclude that there are several ways in which the protein amino acid sequence may regulate the oxidation reduction potential and several ways in which the sequence can modify the binding site between cytochrome c and its redox partners. PMID- 6272839 TI - Complete nucleotide sequence of the chicken chromosomal ovalbumin gene and its biological significance. AB - The nucleotide sequence of the entire chicken chromosomal ovalbumin gene has been determined. The gene is 7564 nucleotides in length to code for a mature messenger RNA of 1872 nucleotides. Comparison of the sequence at the 5'-terminal region of the gene with that reported by others has revealed multiple polymorphic nucleotides in the structural, intervening, and flanking DNA sequences. Some of the polymorphic sites occur at positions very close to splice junctions or the eucaryotic promoter sequence, yet apparently have little or no effect on the expression of this gene. The heptanucleotide promoter sequence TATATAT present in the 5'-flanking region of the ovalbumin gene does not occur within the confines of the gene. Nevertheless, multiple Hogness box sequences similar to those found in other eucaryotic genes were delineated within the boundaries of the gene. These internal Hogness box sequences are not used for transcription initiation. Similarly, the hexanucleotide sequence AATAAA common to all eucaryotic messenger RNAs at the 3'-untranslated region occurs seven additional times within the ovalbumin gene. These sites are not used for transcription termination or polyadenylation. Thus, although these sequences may play important roles in the initiation or termination of gene transcripts as well as polyadenylation of the transcripts, the specificity for such biological functions must not reside within these sequences alone. Furthermore, sequences complementary to the highly conserved rat U1 small nuclear RNA have been found throughout the gene. Many of these regions of complementarity occur in the structural sequences. If the small nuclear RNA does play a role in splicing, the specificity must be provided also by other as yet undefined components. PMID- 6272841 TI - Transcription of the major Drosophila heat-shock genes in vitro. AB - Active eukaryotic genes are more accessible to some proteins than bind DNA than are inactive genes. In order to probe the accessibility of the Drosophila heat shock genes we have isolated nuclei from Drosophila tissue culture cells and have used these nuclei as templates for Escherichia coli RNA polymerase. With nuclei isolated from cells that had not been heat shocked, the synthesis of heat-shock RNA was not detected by hybridization to a DNA clone containing sequences from the major heat-shock region. In contrast, approximately 0.22% of the RNA synthesized in nuclei isolated from cells that had been previously heat shocked hybridized to the heat-shock clone. The synthesis of heat-shock RNA was DNA dependent, was sensitive to rifampicin and to actinomycin D, and represented a 70 fold enrichment over random transcription of the Drosophila genome. Transcription showed an extraordinary preference for a region 5' distal to the structural gene. These results demonstrate that preferential transcription by the bacterial RNA polymerase is indicative of the active state of Drosophila genes. PMID- 6272842 TI - Phase Equilibria in binary mixtures of dimyristoylphosphatidylcholine and cardiolipin. AB - Paramagnetic resonance spectra of the spin-label 2,2,6,6-tetramethylpiperidinyl-l oxy have been used to study phase separations in binary mixtures of dimyristoyl phosphatidylcholine and cardiolipin. Two different samples of cardiolipin were used: (i) One sample contained calcium ions at a mole ratio of calcium:cardiolipin = 1:2; the experimental data support the view that cardiolipin is present in the bilayer membrane as calcium ion linked dimers, (CL)2 Ca2+. (ii) A calcium-free sodium cardiolipin sample yielded remarkable spin label partition data that were quite different from those obtained in the presence of Ca2+. In both cases the spin-label data provide evidence for compound formation and for fluid-fluid immiscibility in the bilayer membrane. PMID- 6272843 TI - Nucleotide sequences of 5' termini of adenovirus 2 early transforming region E1a and E1b messenger ribonucleic acids. PMID- 6272844 TI - Structure of chromatin at deoxyribonucleic acid replication forks: prenucleosomal deoxyribonucleic acid is rapidly excised from replicating simian virus 40 chromosomes by micrococcal nuclease. AB - Replicating simian virus 40 (SV40) chromosomes were found to be similar to other eukaryotic chromosomes in that the rate and extent of micrococcal nuclease (MNase) digestion were greater with replicating than with nonreplicating mature SV40 chromatin. MNase digestion of replicating SV40 chromosomes, pulse labeled in either intact cells or nuclear extracts, resulted in the rapid release of nascent DNA as essentially bare fragments of duplex DNA (3-7S) that had an average length of 120 base pairs and were degraded during the course of the reaction. In addition, nucleosomal monomers, equivalent in size to those from mature chromosomes, were released. On the other hand, MNase digestion of uniformly labeled mature SV40 chromosomes resulted in the release of only nucleosomal monomers and oligomers. The small nascent DNA fragments released from replicating chromosomes represented prenucleosomal DNA (PN-DNA) from the region of replication forks that encompasses the actual sites of DNA synthesis and includes Okazaki fragments. Predigestion of replicating SV40 chromosomes with both Escherichia coli exonuclease III (3'-5') and bacteriophage T7 gene 6 exonuclease (5'-3') resulted in complete degradation of PN-DNA. This result, together with the observation that isolated PN-DNA annealed equally well to both strands of SV40 restriction fragments, demonstrated that PN-DNA originates from both sides of replication forks. Over 90% of isolated Okazaki fragments annealed only to the retrograde DNA template. The characteristics of isolated PN-DNA were assessed by examining its sensitivity to MNase and single strand specific S1 endonuclease, sedimentation behavior before and after deproteinization, buoyant density in CsCl after formaldehyde treatment, and size on agarose gels. In addition, it was observed that MNase digestion of purified SV40 DNA also resulted in the release of a transient intermediate similar in size to PN-DNA, indicating that a DNA protein complex is not required to account for the appearance of PN-DNA. These and other data provide a model of replicating chromosomes in which DNA synthesis occurs on a region of replication forks that is free of nucleosomes and is designated as prenucleosomal DNA. PMID- 6272845 TI - Spectral studies of cobalt (II)- and Nickel (II)-metallothionein. AB - The zinc and cadmium of native rabbit metallothionein-1 were replaced stoichiometrically with either cobalt (II) or nickel (II). The electronic, magnetic circular dichroic (MCD), and electron spin resonance spectra of Co (II) metallothionein reflect distorted tetrahedral coordination of the cobalt atoms. Both the d-d and charge-transfer spectral regions closely resemble those of simple cobalt-tetrathiolate complexes, implying that their coordination chemistry is analogous. Ni (II) complex ions and Ni (II)-metallothionein similarly exhibit analogous MCD bands in the d-d region. The circular dichroic bands associated with ligand-metal charge-transfer transitions in the non-d-d region of Co (II)- and Ni (II)-metallothionein afford additional evidence for the similarity in tetrahedral microsymmetry of the two metal derivatives. The known ratio of 20 thiolate ligands to 7 metal ions, in conjunction with the spectral evidence for tetrathiolate coordination in metallothionein, represents good evidence that these metal thiolates are organized in clusters. PMID- 6272846 TI - Immunochemical evidence for a transmembrane orientation of both the (Na+, K+) ATPase subunits. AB - Antibodies were raised against the large catalytic subunit (apparent Mr 96000) and the glycoprotein (apparent Mr 60000) of the sodium- and potassium-dependent adenosine triphosphatase [(Na+, K+)-ATPase] from Bufo marinus. The specificity of each antiserum was assessed by two-dimensional immunoelectrophoresis using toad kidney microsomes or the purified holoenzyme as a source of antigen and by indirect immunoprecipitation of detergent-solubilized (Na+, K+)-ATPase subunits from radioiodinated or biosynthetically labeled kidney holoenzyme, microsomes, or postnuclear supernatant. The anticatalytic subunit serum reacted exclusively with a 96000-dalton protein. The antiserum to the glycoprotein was rendered specific to this subunit by absorption with purified catalytic subunit. The two antisera were agglutinating and lytic in the presence of complement when toad erythrocytes were used as targets, indicating that antigenic determinants of both subunits were exposed on the cell surface. The specific reactivities with surface-exposed antigenic determinants of both subunits could be absorbed with toad red blood cells. Such absorbed antisera still reacted with detergent-treated or untreated kidney microsomes, revealing the presence of cytoplasmic and/or intramembranous antigenic sites. Our immunochemical data demonstrate that the glycoprotein subunit of (Na+, K+)-ATPase spans the lipid bilayer and confirm the transmembrane orientation of the catalytic subunit postulated from functional studies. PMID- 6272847 TI - Function of the iron-sulfur protein of the cytochrome b-c1 segment in electron transfer and energy-conserving reactions of the mitochondrial respiratory chain. PMID- 6272849 TI - Rat enterocyte Na+ transport in vitro. Action of parathyroid hormone and calcitonin. AB - Parathyroid hormone (PTH) and calcitonin exert well known effects on the renal tubule which are thought to involve specific hormone receptors and adenyl cyclase. In the intestine, it is not clear whether the action of PTH and calcitonin is only indirect or also direct, and their mechanisms of action are much less well established. In the present study, possible direct effects of PTH and calcitonin on Na+ transport in isolated intestinal epithelial cells of rats were investigated. In the presence of bovine PTH (1.2 I.U/ml) in the incubation medium, the Na+ efflux rate constant (oKNa) of isolated enterocytes was significantly reduced when compared to that in control experiments with the hormone vehicle only. The mean depression of oKNa induced by bovine PTH was 26% as compared to the control (100%) and to that induced by ouabain (4.0 mM) which was 44%. No depressant effect of bovine PTH on oKNa was observed when the isolated enterocytes were incubated with ouabain (4.0 mM). Thus, bovine PTH appeared to inhibit the ouabain-sensitive Na+ pump. When incubating the isolated epithelial cells in an EGTA-containing CA2+-free medium, bovine PTH lost its capacity to inhibit oKNa. Thus, the presence of extracellular Ca2+ appeared necessary for the inhibitory effect of bovine PTH. In contrast to its effect on oKNa, bovine PTH induced no change in net Na+ uptake by isolated enterocytes. Moreover, no significant effect on enterocyte Na+ transport could be demonstrated for salmon or porcine calcitonin at two different concentrations in the incubation medium, Neither bovine PTH nor salmon calcitonin induced significant changes in enterocyte cyclic AMP or cycle GMP concentrations. It was concluded that bovine PTH, but not calcitonin, exerted a directed inhibitory effect on the ouabain-sensitive oKNa of isolated rat enterocytes. The effect of bovine PTH occurred without measurable activation of the cyclic nucleotide system but needed the presence of Ca2+ in the incubation medium to be operative. PMID- 6272848 TI - Ubiquinone-binding proteins. PMID- 6272850 TI - Effects of temperature, lipid modification and pH on the mobility of the major proteins of the receptor-rich membranes from Torpedo marmarata. AB - The factors influencing the overall mobility of the major proteins of the acetylcholine receptor-rich membranes from Torpedo marmorata have been investigated by saturation transfer ESR spectroscopy and the lateral distribution of these proteins has been studied by electron microscopy. A spin-labelled derivative of maleimide, 3-maleimido-2,2,5,5-tetramethyl-1-pyrrolidinyloxyl (MSL), was used under various conditions of incubation, enabling us to attach it mainly to either an extrinsic protein of 43 kdaltons, or an intrinsic protein (40 kdaltons) bearing the alpha-toxin-binding site. (1) The direct reaction of MSL with the membrane fragments resulted in almost exclusive labelling of the 43 kdalton protein, an extrinsic protein located on the inner face of the receptor rich membranes. (2) After the free SH groups were blocked with N-ethylmaleimide and the disulfide bridges opened with the reducing agent dithiothreitol, MSL reacted with both the 40 and 43 kdalton proteins (6.0 +/- 0.6 MSL molecules per alpha-toxin-binding site). (3) After the latter labelling procedure membranes were exposed to pH 11, resulting in extraction of the 43 kdalton protein and leaving 2.2 +/- 0.4 MSL molecules per alpha-toxin-binding site; sodium dodecyl sulfate polyacrylamide gel electrophoresis performed with N-[14C] ethylmaleimide suggested that MSL was bound mainly to the 40 kdalton polypeptide chain of the acetylcholine receptor. The following conclusions were made with the native and alkaline-treated membranes: In the native membranes, saturation transfer ESR does not reveal any significant protein rotational diffusion (rotational correlation time tau C greater than 1 ms). Temperature variations and/or lipid modifications obtained by fusion of exogenous lipids and/or cholesterol exchange have little influence on the saturation transfer ESR spectra. Electron microscopy reveals that upon lipid addition, proteins remain in the form of clusters while areas depleted of proteins appear. On the other hand, alkaline treatment strikingly enhances the motion of the MSL-labelled proteins in the membrane (100 less than or equal to tau c less than or equal to 120 microseconds). Furthermore, the rotational diffusion of the MSL-labelled proteins (mainly the 40 kdalton protein) becomes sensitive to temperature, lipid composition and the lipid-to-proteins ratio. Electron microscopy shows that alkaline extraction does not cause large reorganization of the acetylcholine receptor in the plane of the membrane. However, when phospholipids are added to pH 11 treated membranes, a dispersion of the receptor and rosettes is observed. In contrast, cholesterol enrichment of the latter membranes induces clustering of the receptor immobilization as judged by saturation transfer ESR. Upon reassociation of the pH 11 soluble proteins with the alkaline-treated membranes, the restriction of the acetylcholine receptor rotational mobility is also restored (tau c greater than or equal to 1 ms). PMID- 6272851 TI - Effects of secretagogues on the K+ permeability of mucosal and serosal borders of rabbit colonic mucosa. AB - (1) K+ efflux rates from the mucosal and serosal surfaces of sheets of rabbit colonic mucosa have been determined by measuring net K+ loss into K+-free Ringer solution bathing each side of the tissue. (2) Initially, there is a high rate of K+ loss from the tissue, this falls to a lower steady-state rate after 20 min. Loss of K+ from the tissue into the serosal bath is 6-8 fold faster than loss to the mucosal bath. (3) A number of intestinal secretagogues, e.g. theophylline, cyclic AMP, carbachol, ionophore A23187, as well as the laxative bisacodyl, raise the K+ efflux rate across the mucosal border by 200-300%. In the case of K+ efflux induced by carbachol the effect is shown to be dependent on raised levels of intracellular Ca2+. Ca2+-calmodulin complex does not appear to be be involved in activation of K+ efflux across the mucosal border. (4) Amiloride does not block mucosal K+ efflux, but tetraethyl-ammonium does inhibit K+ efflux across the mucosal border, induced by either bisacodyl or raised intracellular Ca2+. (5) The results suggest that laxatives may increase the rate of K+ secretion into the colonic lumen by raising the K+ permeability of the mucosal border. PMID- 6272852 TI - The steady-state kinetic mechanism of ATP hydrolysis catalyzed by membrane-bound (Na+ + K+)-ATPase from ox brain. AB - The expressions for the kinetic constants corresponding to the steady state model for hydrolysis of ATP catalyzed by (Na+ + K+)-ATPase proposed recently are analyzed with the object of determining the rate constants. The theoretical background for the necessary procedures is described. The results of this analysis are: (1) A small class (four) of rate constants are determined directly by the previously published values of the kinetic constants. (2) For a somewhat larger class of rate constants upper and lower bounds may be established. For several rate constants the upper and lower bounds differ by less than a factor 1.6 (for the "(Na+ + K+)-enzyme", i.e. the enzyme activity with K+ and millimolar substrate concentration) and 1.2 (for the "Na+-enzyme",i.e. the activity at micromolar substrate concentrations). (3) Experiments on inhibition by K+ of the Na+-enzyme at various Mg2+ concentrations are reported and analyzed. With the additional assumption that the rate constants governing the addition to ATP of Mg2+ is independent of whether or not ATP is bound to an enzyme molecule, a set of consistent values for all the 23 rate constants in the mechanism may be obtained. (4) The values of some rate constants lend further support to the contention discussed in a previous paper that the enzyme hydrolyzes ATP along two kinetically distinct pathways, depending on the presence of K+ and on the concentration of substrate, without the necessity of having more than one active substrate site per enzyme unit at any time. (5) The results show that while the two enzyme forms, the "Na+-enzyme" E1 and the "K+-enzyme" E2K, add substrate with (second order) rate constants of the same order of magnitude (differing only by a factor of four in favor of the former), the rate constants for the reverse processes differ by a factor of 100, being largest for the K+-enzyme. This is the main reason for the large difference in the Michaelis constants for the two forms reported previously. (6) Compatibility of the model with the well-known rapid dephosphorylation of the phosphorylated enzyme in the presence of K+ requires the presence, at non-zero steady state concentration, of an enzyme-potassium phosphate intermediate, which is acid labile and is therefore not detected as a phosphorylated enzyme using conventional methods. PMID- 6272854 TI - Radiation inactivation of (Na+ + K+)-ATPase. A small target size for the K+ occluding mechanism. AB - Radiation inactivation of partially purified (Na+ + K+)-ATPase (ATP phosphohydrolase, EC 3.6.1.3) from pig kidney outer medulla shows that the target size for Rb+ occlusion by the enzyme (in the absence of phosphorylation) is much smaller than the target size for p-nitrophenyl phosphatase activity, which is itself smaller than the reported target size for (Na+ + K+)-ATPase activity. PMID- 6272853 TI - Cellular pH and water permeability control in frog urinary bladder. A possible action on the water pathway. AB - Mucosal acidification (from pH 8.1 to 6.0) reversibly inhibited the hydroosmotic responses to oxytocin, cyclic AMP and 8-bromo-cyclic AMP in frog urinary bladder. These inhibitory effects were only observed in the presence of a permeant buffer in the apical medium and could also be elicited by CO2 bubbling, even when the mucosal pH was clamped at 8.1. Acid pH reduced the oxytocin-induced net water flux faster than norepinephrine or oxytocin removal and the difference was especially important at low temperature. The time course of recovery from acid pH inhibition was, at 20 degree C, similar to that of the hormonal action, but when the medium temperature was reduced to 6-7 degrees C, the recovery from acid pH inhibition paradoxically became faster while the oxytocin action was markedly slowed down (t 1/2 of changes in net water fluxes (expressed in min): oxytocin addition at 20 degrees C, 6.2 +/- 0.9; at 6 degrees C, 24 +/- 3; oxytocin removal at 20 degrees C, 4.7 +/- 0.8; at 6 degrees C, 22 +/- 3; pH inhibition at 20 degrees C, 2.6 +/- 0.2, at 6 degrees C 2.5 +/- 0.2; recovery from pH 6 at 20 degrees C 6.5 +/- 0.9; at 6 degrees C, 2.7 +/- 0.3). These results can be explained by accepting two main loci sensitive to medium acidification: (1) the cyclase system and (2) an intracellular, temperature-independent, post-cyclic AMP site. The fact that the intramembranous particle aggregates associated with the oxytocin-induced water permeability increase did not disappear after the flow inhibition by acid pH at low temperature suggests that the second effect could be located at the water channel itself. PMID- 6272855 TI - Studies on (K+ + H+)-ATPase. IV. Effects of phospholipase C treatment. AB - (1) The total phospholipid content of a gradient purified (K+ + H+)-ATPase preparation from pig gastric mucosa is 105 mumol per 100 mg protein, and consists of 29% sphingomyelin, 29% phosphatidylcholine, 28% phosphatidylethanolamine, 10% phosphatidylserine and 4% phosphatidylinositol. The cholesterol content corresponds to 50 mumol per 100 mg protein. (2) Treatment with phospholipase C (from Clostridium welchii and Bacillus cereus) results in an immediate decrease of the phosphate content. Up to 50% of the phospholipids are hydrolyzed by each phospholipase C preparation alone, without further hydrolysis by increased phospholipase concentration or prolonged incubation time. Combined treatment with the two phospholipase C preparations, sequentially or simultaneously, hydrolyzes up to 65% of the phospholipids. (3) The (K+ + H+)-ATPase and K+ stimulated p nitrophenylphosphatase activities are decreased proportionally with the total phospholipid content, indicating that these enzyme activities are dependent on phospholipids. (4) Phospholipase C treatment does not change optimal pH, Km value for ATP and temperature dependence of the gastric (K+ + H+)-ATPase, but slightly decreases the Ka value for K+. (5) Phospholipase C treatment lowers the AdoPP[NH]P binding and phosphorylation capacities, suggesting that inactivation occurs primarily on the substrate binding level. (6) Most of the results can be understood by assuming that hydrolysis of the phospholipids by phospholipase C leads to aggregation of the membrane protein molecules and complete inactivation of the aggregated ATPase molecules. PMID- 6272856 TI - The labeling with 8-azido-cyclic adenosine monophosphate of proteins in vesicles of sarcoplasmic reticulum from rabbit skeletal muscle. AB - Photoinduced labeling with 8-azido-cyclic AMP of proteins in vesicles from sarcoplasmic reticulum rabbit skeletal muscle has been examined. At concentrations of 0.1 microM or less, specific labeling of three or four bands, representing trace components in the SDS gels, was seen. This labeling was prevented by cyclic AMP. Some of the bands correspond approximately in apparent molecular weight to subunits of cyclic AMP-dependent protein kinases reported in other systems. Attempts to use this reagent as a probe for protein position in the membrane have been unsuccessful since 8-azido-cyclic AMP passes into and through the membrane. The reagent also appears to partition with a linear concentration dependence into the vesicular membrane itself, presumably into the lipid portion. PMID- 6272857 TI - Stimulation of D-glucose transport. A novel effect of vitamin D on intestinal membrane transport. AB - Vitamin D stimulates absorption of D-glucose in chick jejunum and ileum by a specific action on the maximal velocity of Na+-gradient driven D-glucose transport across the brush-border membrane of intestinal cells. Induction of D glucose transport by either vitamin D-3 or 1,25-dihydroxyvitamin D-3 in embryonic intestine can be blocked by inhibitors of RNA and protein synthesis. PMID- 6272858 TI - The abnormal phosphorylation of spectrin in human hereditary spherocytosis. AB - The phosphorylation of the proteins of the erythrocyte membrane of patients suffering from hereditary spherocytosis is investigated in intact erythrocytes by their incubation in the presence of radioactive inorganic phosphate. Examination of the phosphorylated components by high-resolution two-dimensional gel electrophoresis reveals only one defect in the pathological membranes, a depressed phosphorylation of the smaller polypeptide of spectrin; band 2. The phosphorylation of band 2 is measured with reference to the phosphorylation of syndein (2.1 + 2.2 + 2.3). In patients showing overt clinical symptoms and for whom splenectomy is advocated the phosphorylation of band 2 is depressed by approx. 70%. After splenectomy the phosphorylation of membrane proteins is restored to normal levels. PMID- 6272859 TI - Interaction of superoxide dismutase with phospholipid liposomes. An uptake, spin label and calorimetric study. AB - The effect of superoxide dismutases from five species upon phospholipid bilayers has been investigated. The uptake by egg phosphatidylcholine bilayers of the holo and apo forms of bovine superoxide dismutase increases with enzyme concentration and only a fraction of each is removed by treatment with trypsin. These uptake data indicate that both forms of the enzyme associate with and are embedded within lipid bilayers. From the spectrum of the spin label 2-(3-carboxypropyl) 4,4-dimethyl-2-tridecyl-3-oxazolidinyloxyl, the binding of superoxide dismutase to egg phosphatidylcholine bilayers can be shown to disorder the lipid packing. The disordering by the bovine holoenzyme is small but increases with increasing enzyme concentration and period of incubation. The disordering effects of the apoenzyme are much larger and are reversible by Cu2+, Zn2+ reconstitution of the apoenzyme. The disordering effect of the apoenzyme is further confirmed by differential scanning calorimetry. The gel to liquid crystalline phase transition of egg phosphatidylcholine is lowered 7 degrees C by 25% by weight apo-superoxide dismutase to lipid. Human, dog, swordfish and yeast superoxide dismutases also disorder, and to a greater extent than the bovine enzyme. The greatest perturbation is produced by yeast superoxide dismutase; a 20% decrease in the order parameter by 50% by weight enzyme to lipid. PMID- 6272860 TI - The relationship between plasma membrane lipid composition and physical-chemical properties. II. Effect of phospholipid fatty acid modulation on plasma membrane physical properties and enzymatic activities. AB - The fatty acid composition of plasma membrane phospholipids of the murine T lymphocyte tumor EL4 were systematically modified in an attempt to understand the relationship between lipid bilayer composition and plasma membrane physical and biological properties. Two plasma membrane enzyme activities, adenylate cyclase and ouabain-sensitive (Na+ + K+)-ATPase, were measured in normal and fatty acid substituted EL4 plasma membrane fractions. The fatty acid effect on enzyme activities was similar to previously reported effects of fatty acids on cytotoxic T cell function. The activity of both enzymes was inhibited by saturated fatty acids, while unsaturated fatty acids had a moderate enhancing effect on both enzyme activities. Using two different nitroxide derivatives of stearic acid, the order parameter and approximate rotational correlation times were calculated from ESR spectra of normal and fatty acid-modified plasma membranes. No significant differences was found in either parameter in these membranes. These results, in conjunction with earlier data from our laboratory and others, suggest that caution should be exercised in inferring changes in membrane 'fluidity' based on lipid modulation of biological membranes. PMID- 6272861 TI - Triiodothyronine receptor from rat liver nuclei. Interaction, after partial purification, with DNA and chromatin. PMID- 6272862 TI - DNA fork displacement rates in human cells. AB - DNA fork displacement rates were measured in 20 human cell lines by a bromodeoxyuridine-313 nm photolysis technique. Cell lines included representatives of normal diploid, Fanconi's anemia, ataxia telangiectases, xeroderma pigmentosum, trisomy-21 and several transformed lines. The average value for all the cell lines was 0.53 0.08 mum/min. The average value for individual cell lines, however, displayed a 30% variation. Less than 10% of variation in the fork displacement rate appears to be due to the experimental technique; the remainder is probably due to true variation among the cell types and to culture conditions. PMID- 6272863 TI - Characterization of the Epstein-Barr virion-associated DNA polymerase as isolated from superinfected and drug-stimulated cells. AB - We reported previously that Epstein-Barr (EB) virions and detergent-treated nucleocapsids co-purified with significant amounts of DNA polymerase activity that did not resemble other known host or viral polymerases. We report here that this species of DNA polymerase activity is present at early times after infection in lymphocytes abortively lytically infected (superinfected) with EB virus. However, studies with [35S]methionine labeling suggest de novo synthesis of enzyme has not occurred. Conversely, drug-stimulated lymphocytes that synthesize EB viral late proteins and virions contain this species of polymerase to the virtual exclusion of all others. This EB viral polymerase shows a marked preference for nicked and gapped double-stranded rather than primed single stranded DNA templates. Its processiveness as measured on primed theta X174 phage DNA template is lower than that of lymphocyte beta polymerase. The data reported here are consistent with the hypothesis that the EB virion-associated DNA polymerase is synthesized at late times in the viral life cycle as are other structural proteins but it plays an important role early after viral infection. It is known that mature herpes virion DNA (including that of EB virus) is nicked and gapped and we propose that virion polymerase repairs the viral DNA at an early stage in infection before viral DNA replication begins. PMID- 6272864 TI - Sequence sensitivity of histone binding. AB - The nucleotide sequence selectivity of histone binding has been measured by thermal denaturation of reconstituted nucleoproteins. When DNAs of different average base compositions competed for the binding of purified histone fractions during in vitro reconstitutions in the presence of salt and urea, a decreasing (A + T)-binding preference was observed following the order H1 greater than H2B greater than H5 greater than H2A greater than [H2A + H2B] greater than [H2A + H2B + H3 + H4], [H1 + (H2A + H2B + H3 + H4)2]. Nucleoprotein complexes formed under conditions shown to yield more physiologically comparable nucleosome structures revealed a minimal (A + T)-binding preference. These results suggest that homotypic and heterotypic histone interactions decreased the nucleotide sequence selectivity of nucleosome binding. PMID- 6272865 TI - Substrate-induced intramolecular proton transfer in 6-phosphogluconate dehydrogenase from Candida utilis. AB - Formation of binary complex between 6-phosphogluconate dehydrogenase (6-phospho-D gluconate:NADP+ 2-oxidoreductase (decarboxylating), EC 1.1.1.44) from Candida utilis and 6-phosphogluconate was investigated by means of ultraviolet difference spectroscopy. The formation of the enzyme-substrate complex induces in the difference spectrum a positive peak the wavelength and extinction coefficient of which agree well with a tyrosine ionization. Titrimetric studies indicate that the formation of the binary complex is not coupled to a proton release from the protein. These data support an intramolecular proton transfer from a tyrosine to other functional group. This proton transfer could be correlated to the conformational change induced by substrate in 6-phosphogluconate dehydrogenase. PMID- 6272866 TI - Aminopeptidase A in human placenta. PMID- 6272867 TI - Characterization of pancreatic islet Ca2+-ATPase. AB - Ca2+-dependent ATPase (Ca2+-dependent ATP phosphohydrolase, EC 3.6.1.3) present in a subcellular fraction derived from rat pancreatic islet homogenates was examined to determine kinetic parameters and responses to various substances with known effects upon insulin secretion. Experiments demonstrated the presence of a Ca2+-ATPase with a Km ATP of 7 . 10(-5) M and two Km Ca of 1.3 . 10(-7) M and 5.7 . 10(-6) M. The enzyme had little activity in acidic media while retaining considerable activity in basic media. Optimal activity was obtained at pH 7.5. The enzyme was relatively temperature insensitive (Q10 = 1.49), since activity decreased less than 50% with a 15 degrees C decrease in temperature. Studies on the stability of enzyme activity upon storage at -20 degrees C indicated that for intact islets activity was stable for 3 weeks, while in homogenates activity was stable for only 1 week, after which activity rapidly declined in both cases. Certain substances known to either stimulate or inhibit insulin secretion were tested for their ability to alter enzyme activity. Potassium, glibenclamide and cyclic AMP had no effects upon activity. These observations are consistent with the hypothesis that a Ca2+-ATPase present in pancreatic islets may act as a modulator of pancreatic islet beta cell activity. PMID- 6272868 TI - Synthetic fragments of protamines as model substrates for rat liver cyclic AMP dependent protein kinase. PMID- 6272869 TI - Effects of insulin on lipolysis and lipogenesis in hamster white adipocytes with high sensitivity to hormones. AB - A modified procedure for preparation of hamster adipocytes by collagenase digestion under carefully controlled conditions has been developed. The adipocytes were 4- to 8-fold more sensitive to catecholamine stimulation of lipolysis than cells prepared by a commonly used method (Hittelman, K.J., Wu, C.F. and Butcher, R.W. (1973) Biochim. Biophys. Acta 304, 188-196) and also more sensitive to the anti-lipolytic action of insulin. The effects of insulin on lipogenesis, measured as [3H]glucose conversion to cell lipids, and on catecholamine-stimulated lipolysis were compared under identical conditions with the same cell batch. Isoprenaline-stimulated lipolysis was found to be half maximally inhibited by an insulin concentration 8-fold lower than that stimulating lipogenesis to a corresponding extent (half-maximal effects at insulin concentrations of 40 vs. 300 pM). A similar difference was found when cells had been stimulated with adrenaline instead of isoprenaline. PMID- 6272870 TI - Lipoprotein apolipoprotein synthesis by human hepatoma cells in culture. AB - Lipoprotein synthesis was demonstrated by double diffusion with low density lipoprotein antibody, and by 3H-labeled amino acid incorporation into proteins of the d less than 1.063 g/ml centrifugally isolated lipoprotein fraction. Radioactive label was incorporated predominantly into apolipoprotein B (60%), apolipoprotein A-I (20%) and apolipoprotein C (12%), as determined by Sepharose column chromatography and polyacrylamide gel electrophoresis. Incorporation of radioactive label into apolipoprotein B was inhibited by the presence of albumin in the medium, and was restored to control levels with the addition of 1 mM oleic acid, indicating that cell synthesis of apolipoproteins could be modified by culture conditions. The human hepatoma cell line, Hep G2, provides a potential in vitro model for the study of regulation of human hepatic lipoprotein and apolipoprotein synthesis. PMID- 6272871 TI - Analogues of cyclic AMP inhibit phosphatidylethanolamine N-methylation by cultured rat hepatocytes. PMID- 6272872 TI - Spatial separation of the two essential thiol groups and the binding site of the exchangeable GTP in brain tubulin. A spin label study. AB - The assembly of microtubules from tubulin prepared without glycerol was inhibited by blocking the two most reactive sulfhydryl groups of the eight free sulfhydryl groups present per tubulin dimer. The assembly was also inhibited by Cu2+ ions in a redox-reaction with the two most reactive sulfhydryl groups. These two sulfhydryl groups had almost the same reactivity towards N-ethylmaleimide and p chloromercuribenzoate, in spite of the fact that they are located on different subunits of tubulin. It was not possible to label just one single sulfhydryl group at a time by N-ethylmaleimide, and it was not possible to decide whether one or two free sulfhydryl group(s) are needed for assembly. The EPR technique based on the interaction of spin labels with transition metals was used for the study of the distance between the two most reactive sulfhydryl groups and the sites of exchangeable GTP and Mg2+, respectively. The sulfhydryl groups were spin labelled with a nitroxide derivative of N-ethylmaleimide. Cr(III)GTP was used as a paramagnetic substitute for GTP, and Mn2+ for Mg2+. It was found that: a. The exchange of GTP and the total content of GTP were not affected by modification of the sulfhydryl groups. b. The binding sites of the exchangeable GTP and Mg2+ are located 10 A, at least, from the two most reactive sulfhydryl groups. c. The distance between the spin labels introduced on the two most reactive sulfhydryl groups was larger than 17 A. The findings indicate that there is no direct interaction between exchangeable GTP and the two most reactive sulfhydryl groups. PMID- 6272873 TI - The denatured states of cytochrome c. AB - The unfolding of horse ferricytochrome c in the presence of several inorganic salts has been studied under a variety of denaturing conditions and followed by means of absorbance changes in the Soret region (390-450 nm) and visible region (450-750 nm), as well as by viscosity measurements. Change in the Soret region, usually sensitive to the heme environment, were characterized by gradual increases in absorbance at 409 nm for low concentrations of NaClO4 and LiClP4. As denaturant concentrations were increased, the low-spin state of the ferric heme is altered, as seen by a maximum shift to shorter wavelengths (402-407 nm) accompanied by a further increase in absorbance in the Soret region. Unlike the effect of several organic denaturants and the above salts, denaturation in the presence of LiCl and CaCl2 resulted in an overall decrease in the Soret region with a blue-shift to 401 and 400 nm, respectively. Visible spectra of ferricytochrome c exhibited new bands at 633 nm (9.0 M LiCl) and 636 nm (4.5 M CaCl2) indicative of a change in the spin state of the iron upon displacement of methionine 80. LiCl and LiBr produced intermediate states of protein unfolding with midpoints (D 1/2) at 4.0 and 8.6, and 2.6 and 6.4, respectively. A determination of delta GHU2O and m, the free energy of unfolding in the absence of denaturant and the dependence of free energy of denaturation on denaturant concentration, was used to analyze the relative effectiveness of these denaturants. PMID- 6272874 TI - The effect of growth state on the activity of the protein kinase isoenzymes. An increase in type II cyclic AMP-dependent protein kinase is correlated with growth arrest in G1 phase. AB - Native polyacrylamide gels have been used to resolve protein kinase isoenzymes from cultured cells and the protein kinases have been identified by carrying out phosphorylation reactions in the gel. Following electrophoresis the gels were incubated with histone and [gamma-32P]ATP. The gels were then thoroughly washed and dried down, and the protein kinases were located by autoradiography. Protein kinase activity as measured in the gel system was a linear function of cytosol protein concentration up to about 100 microgram per channel and incorporation of 32P into histone was time dependent. Three bands of protein kinase activity were resolved in cytosol samples from baby hamster kidney (BHK) fibroblasts. The band with the lowest relative mobility utilized histone IIA or casein equally well as substrate protein whereas bands 2 and 3 demonstrated a clear preference for histone. Bands 2 and 3 displayed a relative mobility in electrophoresis that was identical to that observed for cyclic AMP-dependent protein kinases I and II from rat liver. Treatment of cytosol samples with cyclic AMP prior to electrophoresis resulted in the disappearance of cyclic AMP-dependent protein kinases from the gel profile. This method was employed to identify bands 2 and 3 as cyclic AMP dependent protein kinases. The protein kinases in growth-arrested cells were compared with proliferating cells. We have observed a 3.5-fold increase in the activity of Type II protein kinase as the cells arrest growth in G1 phase of the cell cycle. This increase in Type II is correlated with the increase in cells blocked in G1 and a decrease in Type II activity appears to be an early event in permitting cells to leave G1 and resume growth. PMID- 6272875 TI - Binding of vasoactive intestinal peptide and its stimulation of adenylate cyclase through two classes of receptors in rat liver membranes. Effects of 12 secretin analogues and 2 secretin fragments. AB - 1. Vasoactive intestinal peptide (VIP) receptors were identified in crude rat hepatic membranes by 125I-labelled VIP binding and by the ability of VIP to stimulate adenylate cyclase activity. The specificity of these receptors was evaluated by the capacity of secretin, synthetic secretin analogues, and secretin fragments to inhibit 125I-labelled VIP binding and to stimulate adenylate cyclase. 2. The results were compatible with the existence of two classes of VIP binding sites that could be distinguished according to their affinity for VIP and their specificity. High-affinity sites were more specific for VIP as secretin was 175 times less potent than VIP for recognition of these sites while being only 33 times less potent than VIP for recognition of low-affinity sites. 3. Secretin analogues, monosubstituted in position 2, 3, 4 or 6 were less potent than secretin for adenylate cyclase stimulation as well as for the recognition of the two classes of receptors. [Val5]secretin was more potent than secretin and appeared definitely more VIP-like than secretin; [Ala4, Val5] and [D Ala4,Val5]secretin were equipotent to secretin. 4. The fragment secretin (7-27) was unable to recognize VIP receptors and to stimulate adenylate cyclase. The substituted fragment [Gln9,Asn15]secretin (5-27) recognized these receptors with weak potency but could not activate the enzyme. PMID- 6272876 TI - Periodate-induced transformation of human peripheral blood lymphocytes and the resultant oxidation of membrane sialyl, galactosyl and fucosyl residues. AB - Oxidation of human peripheral mononuclear cells with sodium periodate results in lymphocyte activation. Period-date, at optimal mitogenic concentrations, oxidizes membrane sialyl residues (NeuNAc) essentially into the 7 carbon analogue (C7 NeuNAc). Fucosyl and galactosyl residues are also oxidized by periodate, since propane 1,2-diol and glycerol are isolated in acid hydrolysates of lymphocytes oxidized by periodate and reduced by tritiated borohydride. The neuraminidase pretreatment of lymphocytes induces a 40-50% decrease of their response to periodate. Neuraminidase treatment of 108 human peripheral lymphocytes liberated 9.6 microgram NeuNAc (31 nmol), representing 68.5% of the total content. The neuraminidase treatment dramatically enhances the recovery of glycerol in hydrolysates of lymphocytes treated successively with periodate and tritiated borohydride. PMID- 6272877 TI - The kinetoplast of DNA of Trypanosoma gambiense: comparison with the kDNA of Trypanosoma equiperdum. AB - The molecular components of the kinetoplast DNA (kDNA) network of Trypanosoma gambiense have been studied and compared with those of the very closely related species T. equiperdum, previously studied in detail. The kDNA of T. gambiense contains about 80 maxicircles of 20 kilobase pairs and 4000 minicircles of 1 kilobase pairs. The restriction cleavage sites of 7 restriction endonucleases have been mapped on the T. gambiense maxicircle. The majority of these sites were also found in T. equiperdum maxicircles; however their relative positions which are different do not allow us to conclude to relatedness of maps. Maxicircles of the Cairn or of the rolling circle type have been observed and thought to be replicative intermediates. Experiments on renaturation kinetics and hybridization after blotting transfer, show that T. gambiense and T. equiperdum maxicircles have base sequences in common. The T. gambiense minicircles are heterogeneous in base sequence, in contrast to the T. equiperdum minicircles which are homogeneous. The minicircles of the two species have also common base sequences. PMID- 6272879 TI - Restriction map of the cya region of the Escherichia coli K12 chromosome. PMID- 6272878 TI - Proteins that shape DNA. AB - During the last five years, considerable accumulation of data on nucleic acids metabolism leads to the discovery of a number of proteins designed to change the conformation of DNA and to "shape" it. Experimental results emphasize the importance of the conformation and the flexibility of DNA itself in such interactions. The mutual recognition of nucleic acids by proteins may be or not dependent on the nnucleotide sequence and in most cases is accompanied by conformational changes in the proteins involved. Among these are proteins that bind in stoichiometric amounts to DNA, proteins that promote the separation of the two strands in a duplex, and finally proteins that change the topology of DNA. PMID- 6272880 TI - Calcium-ion-activated neutral proteinase of muscle is found in other tissues. PMID- 6272881 TI - [Mechanism of the inhibiting effect of the Cu-tyrosine complex on the hydroxylation reactions]. AB - The Cu-tyrosine complex, a low molecular weight analog of superoxide dismutase, exerts an inhibiting effect on cytochrome P-450. The inactivation of cytochrome Y 450 with its transition to cytochrome Y-420 can cause the inhibition by the Cu2+ Tyr2 complex of dimethylaniline N-demethylation and p-nitroanisol O demethylation. In case of p-hydroxylation of aniline the inhibiting effect of the Cu-tyrosine complex is much more pronounced than its inactivating effect on cytochrome P-450. In the presence of albumin the complex produced no inactivating effect on cytochrome P-450; under these conditions the inhibiting effect of Cu2+ Tyr2 on N- and O-demethylation is removed. In case of aniline p-hydroxylation albumin partly decreases the inhibiting effect of the complex on this reaction. In a soluble system containing isolated cytochrome P-450 and cumole hydroperoxide only the aniline p-hydroxylation reaction was found sensitive to the effect of superoxide dismutase. The data obtained suggest participation of the superoxide radical only in aniline p-hydroxylation but not in the reactions of N demethylation of dimethylaniline and O-dealkylation of p-nitroanisol. PMID- 6272882 TI - [Extracellular RNAase Pch2 of Penicillium chrysogenum 152A: purification, specificity and physico-chemical properties]. AB - Acid RNAase Pch2 was isolated from a filtrate of the cultural fluid of the fungus Penicillium chrysogenum 152A and purified to homogeneity. An analysis of RNAase Pch2 action on RNA and synthetic substrates showed that the enzyme can be attributed to non-specific true ribonucleases (ribonucleate-3'-oligo-nucleotide hydrolase, EC 3.1.4.23). The maximal effect of the enzyme on RNA is observe at pH 4.5 and 55 degree. The RNAase Pch2 is not activated by bivalent metal ions, p chloromercurybenzoate or beta-mercaptoethanol and is reversibly inactivated by 8 M urea. The enzyme molecule consists of 332 amino acid residues; its molecular weight is 36160, the isoelectric point lies at 5.2. PMID- 6272883 TI - [Interaction of SV40 T-antigen with tumor cell chromatin]. AB - The T-antigen of SV40 virus can be found in purified chromatin prepared from virus-induced tumour cells of the Syrian hamster. After treatment of chromatin or isolated nuclei with micrococcal nuclease this protein is detected in the high molecular weight and oligonucleosomal fractions. Data from sedimentation analysis and gel electrophoresis suggest that the T-antigen is predominantly linked with the oligonucleosomal fraction and in a lesser degree with mononucleasomes containing linker DNA and histone H1. A small amount of the T-antigen is found in the mononucleosome complex devoid of histone H1; however, the ratio of the T antigen to DNA in this case is about 30 times less than that in the oligonucleosomal fraction. In order to investigate the nature of T-antigen binding to nucleosomes, the interaction between the T-antigen and nucleosomes from normal rat liver was studied under restricted binding of the antigen to DNA (pH 8.0). The T-antigen was effectively bound to the nucleosomes and coprecipitated with them in 5 mM MgCl2. It was shown that the T-antigen was adsorbed on columns packed with immobilized histones H1 and nucleosomal histones without H1; the former eluted at 0.15 - 0.25 M NaCl, the latter - at 0.35 - 0.5 M NaCl. The possibility of T-antigen interaction with cellular DNA and protein components of chromatin (primarily to H1) is discussed. PMID- 6272884 TI - [Isolation and properties or uridine kinase from Zajdela hepatoma cells]. AB - Uridine kinase (ATP: uridine-5-phosphotransferase, EC 2.7.1.48) was isolated from cytosol of rat Zajdela ascite hepatoma cells by fractionation with ammonium sulfate and gel-filtration on Sephadex G-200. The enzyme has a pH optimum of 7.2 7.8; Km for uridine is 4.8 . 10(-5) M, that for ATP - 1.9 . 10(-4) M. The optimal ratio of ATP of Mg2+ is 2.6. The enzyme activity is inhibited by end products of pyrimidine biosynthesis with Ki for CTP of 6.0 . 10(-4) M and for UTP of 1.2 . 10(-3) M. The Ki values for uridine competitive analogs, i. e. 6 azauridine, 5-bromuridine and 5-azacytidine are equal to 4.0 . 10(-4) M, 1.5 . 10(-3) M and 2.5 . 10(-3) M, respectively. Further purification of the enzyme on Sepharose 4B allowed to obtain the most active, although heterogeneous fractions purified 86-fold, with specific activity of 11.2 mkmole/hour per mg of protein. Using electrofocusing, uridine kinase was found to consist of two major and one minor active fractions with pH of 6.2, 6.7 and 6.35, respectively. Chromatography on DEAE-cellulose DE-32 resulted in two major active fractions of the enzyme, differing in thermal stability and inhibition by CTP. It may be concluded that Zajdela ascite hepatoma cells contain at least two isoforms of uridine kinase. PMID- 6272885 TI - [Chromosome aberration induction by a temperature-sensitive SV40 mutant in mammalian cells]. PMID- 6272886 TI - A fresh look at dopamine. PMID- 6272887 TI - GnRH-induced regulation of GnRH receptor concentration in the phenobarbital blocked hamster. PMID- 6272888 TI - Effects of prostaglandin F2 alpha treatment on LH and dibutyryl cyclic AMP stimulated progesterone secretion by isolated rat luteal cells. PMID- 6272889 TI - Granulosa cell differentiation in vitro: effect of insulin on growth and functional integrity. PMID- 6272890 TI - Inhibition of 123I-labeled human chorionic gonadotropin binding to gonadal receptors by a factor obtained from rat testicular tissue. PMID- 6272891 TI - Regulation by sulfated glycosaminoglycans of the expansion of cumuli oophori isolated from mice. PMID- 6272892 TI - Gonadal receptors: upregulation in response to elevated serum gonadotropin levels. PMID- 6272893 TI - Influence of adrenal steroids upon testosterone secretion by the boar testis. PMID- 6272894 TI - Effects of hCG and cyclic AMP on aromatization in purified Leydig cells of immature and mature rats. PMID- 6272895 TI - Localization of uterine peroxidase activity in estrogen-treated rats. PMID- 6272896 TI - The effect of tetramethylammonium on single sodium channel currents. AB - Voltage-dependent Na conductance of rat myotubes was studied by patch recordings of single-channels. The patches were excised from the cell with the patch electrode, and the cytoplasmic surface was bathed in either CsF or tetramethylammonium (TMA)-F. Inward currents were examined from -20 to -50 mV. In this range Cs and TMA both appeared to be nearly impermeant, but TMA blocked the channel in a voltage-dependent manner. A first-order blocking site was located a maximum of 89% of the way through the membrane field from the cytoplasmic surface. PMID- 6272897 TI - Critical effects from lipid-protein interaction in membranes. II. Interpretation of experimental results. AB - The effects arising from lipid-protein and lipid-cholesterol interaction are discussed within the framework of a general theoretical description presented in the preceding paper. Available experimental results are interpreted, and new experiments are proposed. In the fluid lipid phase proteins and cholesterol increase the lipid orientational order in their neighborhood, in the ordered phase they decrease it. This leads to a decrease of the latent heat at the ordered-fluid transition, which vanishes at a critical concentration of protein or cholesterol. Theoretical predictions for the critical concentrations agree with results from calorimetry. The approach to the critical point is accompanied by an increase of thermal fluctuations of the lipid order and an increase of the lipid response on small perturbations. Thus proteins and cholesterol increase the lipid specific heat, lateral compressibility, permeability, and lateral diffusion on both sides of the phase transition. Notions such as decrease of cooperativity or fluidity due to protein or cholesterol are reviewed in this context. PMID- 6272899 TI - Totally synthetic crystalline ribonuclease A. PMID- 6272898 TI - Binding of multivalent ligands to mobile receptors in membranes. PMID- 6272900 TI - Design of protease inhibitors. PMID- 6272901 TI - Clr and Cls subcomponents of human complement: two serine proteinases lacking the 'histidine-loop' disulphide bridge. AB - The N-terminal amino acid sequence of human C1s b chain has been extended to 52 residues. The histidine residue involved in the charge-relay system is located at position 38, whereas the histidine-loop disulphide bridge is missing. So far, human complement sub-components C1r and C1s are the only known mammalian serine proteinases lacking this disulphide bridge. PMID- 6272902 TI - Adenosine deaminase and ecto-5'-nucleotidase activities in various leukemias with special reference to blast crisis: significance of ecto-5'-nucleotidase in lymphoid blast crisis of chronic myeloid leukemia. AB - Adenosine deaminase (ADA) and ecto-5'-nucleotidase (5'-N) activities were examined in peripheral leukocytes from patients with leukemias, including nine patients with chronic myeloid leukemia (CML) in blast crisis. Four of none cases of CML in blast crisis were myeloid and the remaining lymphoid morphologically. The diagnosis of CML in lymphoid blast crisis was further contributed by the measurement of terminal deoxynucleotidyl transferase (TdT) activity. In all four cases of lymphoid blast crisis and one of myeloid blast crisis, leukemia cells had high 5'-N activity, while there was a little or no detectable activity in those from four cases of myeloid blast crisis and all of CML in chronic phase. ADA activity was high in seven of nine patients with blast crisis. Taken together, leukemia cells from two cases of lymphoid blast crisis had high ADA and 5'-N activities comparable to those in acute lymphocytic leukemia (ALL) cells. In contrast, the enzyme activities of leukemia cells from all but one patient in myeloid blast crisis were in a range similar to acute myeloid leukemia cells. The implications of these findings are as follows: (1) 5'-N may be used as a new biochemical marker of CML in lymphoid blast crisis. (2) Some lymphoid cells of CML in blast crisis have high ADA, 5'-N, and TdT activities and thus are very similar to ALL cells. PMID- 6272903 TI - Evidence that eosinophils catalyze the bromide-dependent decarboxylation of amino acids. AB - Human eosinophils from subjects with or without myeloperoxidase (MPO) deficiency and guinea pig eosinophils are able to decarboxylate L-alanine in the presence of the cationic detergent cetyltrimethylammonium bromide (CTAB) but not in the presence of the nonionic detergent Triton X-100. Instead, both normal human neutrophils and guinea pig neutrophils decarboxylate L-alanine in the presence of either detergent. When the non-bromide-containing cationic detergent cetyltrimethylammonium hydroxide (CTAOH) is used instead of CTAB, the eosinophils from MPO-deficient subjects are unable to decarboxylate L-alanine. Decarboxylation occurs with the combination CTAOH-Br-, but not with the combinations CTAOH-I-, CTAOH-CI-, or CTAOH-F-. Bromide in the absence of CTAOH does not promote decarboxylation. Triton X-100 and deoxycholate are much less effective in promoting decarboxylation in the presence of bromide. L-Lysine and L aspartic acid are decarboxylated to a considerably lower rate than L-alanine in the presence of CTAOH and Br-. It is concluded that the eosinophils can catalyze the bromide-dependent decarboxylation of the apolar amino acid L-alanine in the presence of a cationic detergent. PMID- 6272904 TI - Somatic cell hybrid analyses of hematopoietic differentiation. AB - A differentiated cell expresses an entire set of specialized features. Somatic cell hybridization provides a method to examine control of gene regulation. We studied the expression of tissue-specific features in hybrids between human promyelocytes (HL-60) and human Burkitt's lymphoma cells (P3HR-1). Two hybrid lines, HP-1 and HP-2, and 18 hybrid clones were established and confirmed by karyotype, isozyme, and surface antigen analyses. The hybrids extinguished the 10 myeloid (HL-60) features that we examined including myeloid morphology, histochemistry, and functions that included response to colony-stimulating factor and ability to differentiate to granulocytes or macrophages. In contrast, the hybrids synthesized immunoglobulin and expressed Epstein-Barr nuclear, early, and viral capsid antigens similar to the P3HR-1 lymphoid parental line. Results are contrasted to the findings when P3HR-1 lymphocytes are fused to human erythroid myeloid cells (K562). Taken together, our results suggest that phenotypic differences between human myeloid and lymphoid cells in the hematopoietic lineage involve mutually exclusive programs and may possibly be mediated by the activity of diffusible, transacting molecules. PMID- 6272905 TI - Metabolic heterogeneity of eosinophils from normal and hypereosinophilic patients. AB - Eosinophils, which may be associated with allergic, parasitic, or neoplastic disease, have a potent oxidative burst that may be activated by particulate or soluble stimuli. Eosinophils from normal persons and patients with hypereosinophilia were compared with respect to their ability to produce the active oxygen product, superoxide anion. Normal eosinophils produced large amounts of superoxide anion under resting conditions (0.53 +/- 0.15 nmoles cyto c/10(5) eos/hr) and when stimulated by preopsonized zymosan (0.85 +/0 1.10 nmoles cyto-c/10(5) eos/hr) or phorbol myristate acetate (PMA) (2.38 +/- 0.46 nmoles cyto-c/10(5) eos/hr). Considerable variation was observed in superoxide production by eosinophils from patients with hypereosinophilia. Eosinophils from a group of four patients with hypereosinophilia associated with neoplastic disease produced less superoxide anion than normal eosinophils when stimulated by preopsonized zymosan or PMA (p less than or equal to 0.05). Eosinophils from a group of 5 patients with other causes of hypereosinophilia produced more superoxide anion than normal eosinophils when stimulated by PMA (p less than or equal to 0.01). These studies demonstrate metabolic heterogeneity of eosinophils from patients with hypereosinophilia, and further emphasize that normal eosinophils and eosinophils from hypereosinophilic patients are not functionally equivalent. PMID- 6272906 TI - [Effect of vitamin D deficiency and vitamin D3 therapy on the pig cementum]. PMID- 6272907 TI - [EPR study of bovine enamel samples from the milk and permanent teeth, normal and fluoridated and X- and beta-ray irradiated: Part 2]. PMID- 6272908 TI - Articulations of the ribs: a pictorial review. AB - The joints of the ribs are true articulations and may be involved by local or general affections of the skeleton. Scoliosis, kyphosis, infections, arthritis (local or general types) may result in fibrosis, abnormal configuration, or ankylosis of the joints in question. In certain types of fractures of the dorsal vertebrae, disruption of the contiguous rib articulations may result in fibrosis or bony depositions and secondarily aid in local stabilization of the fracture. Narrowing of the intercostal spaces, as noted in kyphosis, scoliosis, and arthritis of the dorsal region, does not seem to be routinely associated with clinical findings of intercostal neuralgia. PMID- 6272909 TI - Infiltrating angiolipoma. PMID- 6272910 TI - The surgical anatomy of the interosseous ligament of the subtalar joint as it relates to clubfoot surgery. AB - Confusion exists as to the exact anatomical description of the interosseous ligament of the subtalar joint. An effort was made through anatomical dissections to elucidate the exact boundaries of this tiny ligament. It is the authors' belief that in some resistant clubfeet, unless this ligament is transected completely, a full correction of the clubfoot deformity will not be obtained and a less than satisfying result, or a recurrent clubfoot deformity, may be the sequela. PMID- 6272911 TI - The management of gunshot fractures of the extremities. AB - The treatment of 61 fractures of the extremities resulting from low velocity gunshot wounds is reviewed to evaluate the wisdom of the concept of a limited approach to the treatment of civilian gunshot injuries. The great majority of the wounds were treated successfully by superficial care, antibiotics, and immobilization. However, one fracture, after normal healing of the soft tissue wound, was complicated by an infected pseudarthrosis that occurred after two unsuccessful internal fixations, and in another case there was a failed internal fixation but the patient refused further surgery. Our experience with two cases that required formal debridement also suggests that, although the concept of limited wound care for low velocity gunshot wounds is valid, caution is advisable in its application. PMID- 6272912 TI - Tenosynovial chondromatosis of the shoulder. AB - Two cases of tenosynovial chondromatosis of the shoulder are presented, the literature on this rare lesion is reviewed, and the differential diagnosis, diagnosis, and treatment are discussed. Synovial chondromatosis is a benign monoarticular lesion. It may also be found in tendon sheaths de novo and secondary to local extension. The two cases presented show tenosynovial chondromatosis involving both the shoulder joint and extension into the bicipital groove. PMID- 6272913 TI - Injection injuries of the hand. AB - This type of injury is usually the result of an occupational accident, drug abuse, or is iatrogenic in origin. Identification of the injected material, point of entry, and the existence of neurovascular compromise or infection are essential to proper management. Prompt surgical treatment is important. Early motion exercises help minimize joint stiffness and do not seriously delay wound closure. PMID- 6272914 TI - The use of Lotte's nail in the treatment of nonunion of the tibia. AB - Six cases of established nonunion of the tibia were treated successfully with a Lotte's nail. The procedure consisted of an amalgam of well-known techniques previously used in similar cases with varying degrees of success. The authors feel that this method may be a useful addition to the orthopaedists' armamentarium, for it is relatively simple to perform, needs no special equipment, allows early mobilization with weight-bearing, and requires minimal patient cooperation. PMID- 6272915 TI - Osteoblastic metastasis of lymphoepithelioma simulating osteoid osteoma. A case history. AB - An unusual case of solitary osteoblastic bony metastasis at the proximal end of the femur from lymphoepithelioma is presented. Except for the age of the patient, the clinical picture bore a striking resemblance to that of an osteoid osteoma. The patient's belated report of a history of nasopharyngeal carcinoma gave the clue to an accurate diagnosis. PMID- 6272916 TI - A magnetically augmented elbow prosthesis: design and biomechanical evaluation. AB - Magnetically augmented joint prostheses are a relatively new concept. A prototype of an elbow prosthesis with a magnetically constrained axis was designed by the authors and subjected to various tests. It demonstrated greater stability than nonhinged devices. The magnetic bearing reduces the tensile and rotatory forces on the prosthetic fixation that occur with a constrained hinge device, thus preventing loosening and breakage. PMID- 6272917 TI - Biological effects of magnetic fields generated with CoSm magnets. AB - Experiments were conducted to determine whether any observable short-term biological effects were caused by a static magnetic field created with CoSm magnets similar to those used in the elbow prosthesis described on pages 69-80. Tissue culture studies of several cell lines showed no obvious effects on cell growth rate, morphology, or the ability to grow and remain confluent; and no deleterious effects of the magnetic field were evident as a result of an in-vivo study of wound and bone healing in rats. Long-term effects, if any, have yet to be determined. PMID- 6272918 TI - An unusual type of Forestier's disease. AB - Forestier's disease is generally characterized by the presence of heavy osteophytes in the vertebral column, sometimes associated with bony deposits in the ligaments and tendons at the sites of attachment to the bones of the extremities. The authors describe a variation of this syndrome-a patient with symmetrical bony deposits in the ligaments and tendons at the sites of attachment to the bones of the extremities, but with subminimal osteophytes in the vertebral column. PMID- 6272919 TI - Dysplastic and isthmic spondylolisthesis. PMID- 6272920 TI - Inhibition of central angiotensin I conversion by oral captopril. AB - 1. The central action of captopril (D-3-mercapto-methylpropanoyl-L-proline, SQ 14225) in the rat was evaluated by measuring the effect of the orally administered drug on responses to angiotensin I given intracerebroventricularly. 2. The pressor and dipsogenic effects of angiotensin I (20 ng) injected into the third ventricle were reduced by about 50% for 4 h following the oral administration of captopril, 20mg/kg. Both responses returned to pretreatment control values 8 h after captopril administration. 3. Orally administered captopril affects the action of angiotensin I on the central nervous system thus providing an alternative explanation for the hypotensive effect of captopril. This effect if probably due to the direct passage of captopril through the blood brain barrier. PMID- 6272921 TI - A pharmacological classification of drugs of abuse. AB - This paper questions the validity of the distinction between "psychic" and "physical" dependence in relation to drug addiction. The author describes the action of dependence-producing or addictive drugs on the brain at the neuronal and structural level as well as the alterations of arousal, awareness, impairment of sleep, memory, psychomotor performance and sensory perception which are symptomatic of neuropsychological toxicity. Addictive drugs are consumed for the primary pleasurable reward that they produce as a result of their effects on the pleasure reward mechanisms of the brain. These pleasurable sensations will act as reinforcers for repeated administration when the drug effects have worn off and induce a compulsive drug-oriented behaviour. The tolerance, which develops to addictive drugs and the occurrence of withdrawal symptoms, are added reinforcers. The author therefore proposes a pharmacological classification of the most commonly used addictive drugs based on these foregoing effects which may be quantified to a large extent. PMID- 6272922 TI - The addictive potential of cannabis. AB - The author reviews the literature on the dependence potential of cannabis. Case studies and experiments of tolerance to cannabis as well as psychological and physical dependence on cannabis are presented in man and in laboratory animals. Some effects common to both species are also recorded. Although the addictive potential of cannabis is often compared with the addictive potential of alcohol and tobacco, the author concludes that the characteristics of cannabis tolerance are similar to those of opiate dependence. PMID- 6272923 TI - Identification and differentiation of resinous cannabis and textile cannabis: combined use of HPLC and high-resolution GLC. AB - The combined use of high-pressure liquid chromatography (HPLC) and high resolution gas-liquid chromatography (GLC) afforded a means of isolating three substances in samples of cannabis, of determining their retention times in HPLC and in GLC with and without derivation, and of identifying them, by comparison with the data in the literature, with the three major constituents of cannabis: cannabidiol (CBD), delta-9-tetrahydrocannabinol (delta-9-THC) and cannabinol (CBN), without using either control substances or a mass spectrometer. Furthermore, calculation of the peak area ratios (formula: see text); for the different sample varieties can serve as a criterion for the differentiation of resinous cannabis and textile cannabis. PMID- 6272926 TI - Severe peripheral neuropathy complicating legionnaires' disease. PMID- 6272924 TI - Second primary neoplasm in a dysgerminoma patient. PMID- 6272925 TI - Modification of the oral radiation death syndrome with combined WR-2721 and misonidazole. AB - In order to investigate the combined use of radioprotective and radiosensitizing chemicals we have used WR--2721 and misonidazole to study their combined chemical toxicity and radiomodifying effects in our model using the oral radiation death (ORD) syndrome as the endpoint. Our data demonstrate that WR-2721 does protect against the ORD syndrome. Misonidazole does not protect against the ORD syndrome. Misonidazole does not sensitize the normal oral cavity to local irradiation. WR 2721 and misonidazole produce additive toxicity when injected simultaneously and the combination can provide an overall protective response to the ORD syndrome. We conclude that the combination may be potentially useful for the treatment of cancers of the oral cavity. PMID- 6272927 TI - The 1975 foot-and-mouth disease epidemic in Malta. III. serological response of cattle, sheep, goats, and pigs to type O vaccine. PMID- 6272928 TI - Prevalence of equine infectious anaemia (swamp fever) in Guyana. PMID- 6272929 TI - Changes in neuronal transmission in the rat hippocampus during behavior. AB - Rats with implanted stimulating and recording microelectrodes were trained in a straight alley to repeatedly press a bar placed at one end of the alley and to run for water reward available at the other end. Stimulating the commissural input evoked field EPSP and population spike in the CA1 region while no population spike was observed in the dentate area. The amplitude of the CA1 population spike was maximum during running and smallest during drinking. Field potentials in the dentate gyrus changed in an opposite manner (drink greater than groom greater than press greater than run). Perforant path evoked cell discharges in the dentate gyrus which were maximal during drinking and smallest during running. The behavior dependent changes of the evoked potentials covaried with the frequency and power of the simultaneously recorded theta activity. Perforant path stimulation during the negative-going phase of the dentate theta cycle evoked significantly greater granule cell responses than stimuli during the positive-going phase. These observations suggest that the medical septum exerts a potent biasing effect on the efficacy of other afferent to the hippocampus. PMID- 6272930 TI - Cerebral concussion in rats rapidly induces hypothalamic-specific effects on opiate and cholinergic receptors. AB - Conscious male adult rats were concussed by a blow to the occiput with a blunt dart, shot from a spring-loaded pistol. Animals were decapitated within 2 sec, brains quickly removed, several brain regions homogenized in cold buffer and radioreceptor assays carried out. There was a 20% increase in binding of [3H]naloxone and [3H]QNB in the hypothalamus but not in the amygdala, striatum, hippocampus, cortex, midbrain and hindbrain. For animals killed 5-10 sec after concussion, when normal conscious behavior had returned, there were no changes in binding between control and concussed rats. Binding of [3H]spiroperidol and [3H]ouabain was unaffected by concussion. To rule out changes due to non-specific stress, one series of rats received a blow to the lower body. There was no change from control in [3H]QNB binding to hypothalamic tissues by a body blow as compared with the blow to the head. The findings suggests decreased binding of endogenous endorphins and acetylcholine to their respective receptors as a result of concussion. The transient deficit in endogenous transmitter binding may temporarily interrupt hypothalamic circuits concerned with the state of consciousness. PMID- 6272931 TI - Cholecystokinin receptors are decreased in basal ganglia and cerebral cortex of Huntington's disease. AB - Cholecystokinin (CCK) receptors were found to be significantly reduced in basal ganglia and cerebral cortex of post-mortem from Huntington's patients with matched controls. The magnitude of the reduction in CCK binding (69% in basal ganglia, 43% in cerebral cortex) is consistent with the degree of neuronal degeneration in basal ganglia, but suggests a possibly selective loss of CCK receptor-containing neurons in cerebral cortex of Huntington's patients. PMID- 6272932 TI - Dipropylacetate-induced quasi-morphine abstinence behaviour in the rat: involvement of amygdaloid and thalamic structures. AB - Quasi-morphine abstinence behaviour induced by di-n-propylacetate (DPA) in rats is thought to be caused by an increased GABAergic activity in the CNS. Behavioural responses after intracerebral injections of DPA were studied to gain insight into the centre median-parafascicularis (Cm-Pf) resulted in a large number of body shakes and greater locomotor activity when compared to other brain areas. Injection of DPA into the central amygdala (Ac) resulted in an enhanced number of chewing episodes. Administration of bicuculline methiodide (BMI) into the Cm-Pf, 5 min after intraperitoneal administration of DPA, suppressed by body shakes but had only minor effects on horizontal activity, whereas injection of morphine into the same structure suppressed both behavioural symptoms. It is concluded that GABAergic and opioid mechanisms in the Cm-Pf are involved in the DPA-induced behaviour. Injection of BMI into the central amygdala shortly after i.p. injection of DPA resulted in an increase in the number of body shakes, whereas no effect was observed on activity. Morphine applied to this structure slightly potentiated the locomotor activity, but had no effect on the body shakes induced by DPA. The present results suggest a facilitatory role for a GABAergic system in the Cm-Pf on body shakes, while in the central amygdala a GABAergic system exerts an inhibitory influence on this symptom of abstinence. PMID- 6272933 TI - Adenine dinucleotide effects on rat cortical neurones. PMID- 6272934 TI - Age dependent changes in the methylation of rat brain phospholipids. AB - Phospholipid methyltransferase I and II enzymes methylate phosphatidylethanolamine three times to form phosphatidylcholine. The activity of these two enzymes was determined in synaptosome-enriched fractions from rats 1, 3, 7, 15 and 21 months old. The activity of phospholipid methyltransferase I was significantly greater in 7-, 15- and 21-month-old rats than in 1- and 3-month-old rats. In contrast, the activity of phospholipid methyltransferase II did not change with age. These changes in methyltransferase activity with increasing age may be related to changes in beta-receptor function with increasing age. PMID- 6272935 TI - Ethanol and temperature modify motor axon firing patterns. AB - At high temperature a single orthodromic action potential in the excitor (E) axon to the stretcher muscle of the crab Pachygrapsus crassipes provokes the generation of additional spikes in the peripheral E axon branches. This phenomenon exhibits a critical temperature threshold. Bathing preparations in crab salines made up with ethanol at levels as low as 0.5% (109 mM) reversibly decreased the temperature threshold for peripheral spike generation. Higher levels of ethanol resulted in the peripheral generation of E axon spikes at lower temperatures. These results, together with the observation that the temperature threshold is dependent upon the acclimation temperature, indicate that the peripheral generation of action potentials in the E axon is associated with an increase membrane lipid fluidity. PMID- 6272936 TI - Methyl mercury enhances [3H]diazepam binding in different areas of the rat brain. AB - Three days after the acute oral administration of methyl mercury (MeHg), a 27-60% increase in the total number of binding sites for [3H]diazepam was seen in the retina and different areas of the rat brain, with no change, except in the retina, in the apparent dissociation constant for its ligand. In contrast, MeHg failed to change [3H]spiroperidol and [3H]GABA binding in the same areas. Moreover, MeHg decreased cyclic GMP content in the cerebellar cortex. The various possible mechanisms involved in the action of MeHg on benzodiazepine binding are discussed. PMID- 6272937 TI - Muscimol induces gastric acid secretion after central administration. AB - Intracerebroventricular administration of the GABA agonist, muscimol, resulted in a dose-dependent increase in gastric acid secretion in pylorus ligated rats. The maximum increase in gastric acid secretion occurred in the second hour. The GABA antagonist, bicuculline methiodide, reversed the muscimol effect on gastric acid secretion. Intracerebroventricular administration of D-alanine methionine enkephalin, bombesin and calcitonin significantly suppressed the muscimol-induced gastric acid secretion. The present study suggests that the GABAergic system plays a role in the central control of gastric acid secretion. PMID- 6272938 TI - Rapid changes in GABA binding induced by stress in different areas of the rat brain. AB - Rats habituated to handling preceding sacrifice have higher [3H]GABA receptor binding in different brain areas (cerebellum, frontal cortex, caudate nucleus) than naive animals. The increase in GABA binding in handling-habituated rats is due to an increase in the number of receptors (Bmax) with no changes in the affinity of GABA binding for its ligand (Kd). Foot shock causes a sudden fall in GABA binding in handling-habituated rats but does not, or only slightly, in naive ones. The results indicate that stress causes a rapid decrease in GABA receptor binding in the central system and that the GABA binding values which are usually considered as normal are, in fact, values decreased by the handling manoeuvers preceding sacrifice. PMID- 6272939 TI - 3-mercaptopropionic acid inhibits GABA release from rat brain slices in vitro. AB - 3-Mercaptopropionic acid (3MP) (1 mM) inhibited the potassium-evoked release of endogenous GABA from slices of rat hippocampus and cerebral cortex in vitro. This did not appear to be due to an inhibition of GABA biosynthesis, since 3MP failed to affect the basal rate of GABA release or to accelerate the decline in the GABA content of tissue slices during prolonged exposure to 3MP (up to 120 min). 3MP, furthermore, inhibited the potassium-evoked release of [3H]GABA from preloaded brain slices, suggesting a direct inhibitory effect on GABA release. The threshold concentration was approximately 0.1 mM. 3MP at 1 mM failed to inhibit the potassium-evoked release of [3H]5-hydroxytryptamine, [3H]noradrenaline or somatostatin under similar conditions. The ability of 3MP to inhibit GABA release may contribute to the convulsant properties of this substance in vivo. PMID- 6272940 TI - Differing distributions of receptors for morphine and Met5-enkephalinamide in the dorsal horn of the cat. AB - In the dorsal horn of barbiturate-anaesthetized cats evidence was obtained for a different distribution of enkephalin-preferring and morphine-preferring receptors. When ejected electrophoretically from micropipettes, morphine reduced the nociceptive responses of neurones of laminae IV and V when administered in the substantia gelatinosa but not at more ventral sites including sites near cell bodies. By contrast, Met5-enkephalinamide reduced nociceptive responses at all of these sites of administration often with increased potency near cell bodies. It is proposed that morphine-preferring receptors are located mainly near the terminals of primary afferent fibres whereas those for enkephalin are present on these fibres and also on the dendrites and somata of dorsal horn neurones. These dendritic receptors may be located adjacent to sites of termination of nociceptive afferents, a mechanism which would inhibit nociceptive but not other inputs to these neurones. PMID- 6272941 TI - Release and synthesis rates of catecholamines in hypothalamic, limbic and midbrain structures following intraventricular injection of beta-endorphin in male rats. AB - Serum hormone levels and turnover rates of dopamine (DA), norepinephrine (NE) and epinephrine (E) in tissue of pontine midbrain, limbic caudate, and hypothalamic structures were measured following intraventricular injection of 20 microgram beta-endorphin. Turnover of DA was increased in the locus coeruleus and in structures innervated by the nigrostriatal and mesolimbic DA systems while it decreased in the posterior hypothalamus. The NE turnover was reduced in the locus coeruleus and posterior mediobasal hypothalamus. In all tissues examined, beta endorphin treatment had no significant effect on E turnover. Serum levels of prolactin and corticosterone increased following beta-endorphin treatment whereas LH, FSH and TSH levels decreased. The observed effects of beta-endorphin on preoptic and hypothalamic DA and NE turnover rates may explain these hormonal changes. The reduced NE turnover in the locus coeruleus may be induced by increased DA turnover. The observation, that NE turnover in structures innervated by locus coeruleus neurons may go in different direction suggest that NE turnover is regulated independently from the activity of locus coeruleus neurons, hence that the regulation must occur at the terminals by a direct or indirect interaction with endorphinergic neurons. PMID- 6272942 TI - Intracellular recordings from embryonic chick motoneurones in the isolated perfused spinal cord. AB - Potentials of 13- to 18-day chick embryo motoneurones were recorded intracellularly. Action potentials could be evoked by antidromic or direct stimulation of the cells, but the amplitudes were relatively low (less than 40-45 mV) and no overshoot or marked afterpotentials were observed. Pronounced evoked and spontaneous postsynaptic potentials were observed. Both irregular miniature and bursting high-voltage spontaneous postsynaptic depolarizations were present. The association of spontaneous action potentials with the latter type of spontaneous synaptic activity suggests its possible involvement in mechanisms of spontaneous embryonic motility. PMID- 6272943 TI - Increased excitability of hippocampal unmyelinated fibres following conditioning stimulation. AB - Unmyelinated fibres in the stratum radiatum (area CA1) were electrically stimulated in hippocampal slices from guinea pigs. The size of the evoked fibre volley was increased by a preceding conditioning stimulation to the same fibres, provided that the strength of the conditioning stimulus was larger than that of the test pulse. This facilitation was maximal for 20-30 ms interstimulus intervals and had a duration of about 200 ms. The results are compatible with an increased excitability (reduction in threshold) specific for fibres activated by the conditioning stimulation. PMID- 6272945 TI - Development of rabbit hippocampus: physiology. AB - The postnatal development of the CA1 region of rabbit hippocampus was studied using intracellular techniques in the in vitro slice preparation. Recordings from immature hippocampal neurons revealed spiking activity and functional synaptic contacts, even in the newborn animal. Resting potentials and time constants in such cells were similar to those of mature cells; input resistance was higher and action potential duration longer in the immature rabbits. These cell properties reach adult values by 2-3 weeks. Presumed calcium spikes, as well as sodium spikes, were elicited in animals as young as 1 day, so that it was not possible to determine whether calcium or sodium spikes occur earlier. Synaptic potentials recorded in immature CA1 neurons were long duration depolarizing events associated with a large conductance increase. The postsynaptic potentials (PSPs) were shown to be predominantly excitatory in nature, and could be potentiated by repetitive stimulation at slow rates and low intensities. Such stimulation in many cases could trigger seizure-like activity. Inhibitory PSPs in CA1 neurons were rare in animals less than 1-2 weeks old. Increased occurrence of hyperpolarizing inhibitory PSPs was correlated in time with the appearance of interneuron cell types in physiological recordings. These data reinforce the indication from morphological studies that inhibition is late in developing in rabbit hippocampus. PMID- 6272944 TI - Inhibition of primate spinothalamic tract neurons by stimulation in ipsilateral or contralateral ventral posterior lateral (VPLc) thalamic nucleus. AB - The effects of stimulation in the ventral posterior lateral (VPLc) thalamic nuclei on the activity of primate spinothalamic tract neurons were investigated. All 19 cells studied were strongly inhibited by conditioning trains of stimuli delivered to either ipsilateral or contralateral VPLc. Both background discharges and activity evoked by innocuous or noxious cutaneous stimulation were inhibited. PMID- 6272946 TI - Ontogeny of opiate receptors in rat forebrain: visualization by in vitro autoradiography. AB - The embryonic and postnatal ontogeny of opiate receptors in rat telencephalon was mapped by in vitro autoradiographic localization of [3H]naloxone and [3H]enkephalin binding. Opiate receptors marked by naloxone binding first appear at embryonic day 14 in the striatum, rapidly proliferate to adult densities and at the time of birth, gradually become reorganized into the adult heterogeneous pattern of receptor-rich patches surrounded by sparse, diffuse labeling in the rest of the striatum. The enkephalin binding in the striatum appears later in embryonic development and gradually increases in density to form the rather homogeneous adult pattern. Naloxone binding in the paleocortical olfactory areas appears early also, densely within the molecular layer as soon as it is formed at E16. This density is only temporary, as labeling just after birth falls to low adult levels in all areas except portions of the amygdala. Receptors disappear also in the islands of Calleja and the pallidum. Naloxone binding in the septum and neocortex appears gradually in development. The early appearance of striatal and paleocortical [3H]naloxone-labeled opiate receptors and their localization within the subependymal zones suggest that receptors appear on immature neurons before and during migration and, therefore, may influence the intricate patterns of connections that later form. The delayed appearance of the [3H]enkephalin labeled receptors may reflect the dependence of the peptide binding on later developing molecules of adenylate cyclase. PMID- 6272947 TI - Involvement of cyclic AMP in the regulations of lymphokine induced glia cell stimulation. AB - T and B lymphocytes of human or murine origin were found to secrete a factor which increases the DNA and RNA synthesis of cultured glia cells. This factor, termed glia cell stimulating factor (GSF), is released upon stimulation of such immune cells by mitogen or antigen qualifying it as a lymphokine. In this communication we report on the role of cyclic AMP (cAMP) in regulating the effect of GSF on glia cells. Prostaglandin E1 (PGE1), isoproterenol and theophylline were effective in suppressing the GSF-induced increase of the glia cell proliferation. No inhibition of DNA synthesis and no decrease in cell number was observed when testing these substances on glia cells not being activated by GSF. The drugs were found to induce an increase in cAMP concentrations of glia cells. A partial desensitization of the glia cells to these drug induced elevations of cAMP was detected after pretreatment of the glia cell cultures with GSF. It is suggested that stimulated lymphocytes not only release GSF but also low molecular weight proteins such as PGE1 which regulate the effects of GSF on glia cells by activating their adenylate cyclase. PMID- 6272948 TI - Ontogeny of high-affinity GABA and benzodiazepine receptors in the rat cerebellum: an autoradiographic study. AB - High-affinity GABA and benzodiazepine receptors were localized by light microscopic autoradiography in the developing rat cerebellum. [3H]muscimol was used for the labeling of GABA receptors and [3H]flunitrazepam for benzodiazepine receptors. Very low densities of GABA sites were found during the first postnatal week. GABA receptors start increasing linearly at the end of the second week up to adult levels around the fourth postnatal week. The increase in receptor density is concentrated in the developing granule cell layer. Benzodiazepine receptors are present at birth and increases in the density of receptors were observed already during the first postnatal week. Receptor concentrations reached adult values around the third to fourth weeks postnatally. The increase in benzodiazepine receptors in concentrated in the growing molecular layer with little change in the granule cell layer. The immature cell of the external granule layer were characterized by the absence of receptor sites. At least partial association of high-affinity GABA receptors with granule cells and benzodiazepine receptor with Purkinje cell dendrites is suggested by these developmental profiles. PMID- 6272949 TI - Nerve outgrowth, synaptogenesis and bioelectric activity in fetal rat cerebral cortex tissue cultured in serum-free, chemically defined medium. AB - Dissociated and non-dissociated cerebral cortex of fetal rat was successfully cultured in a serum-free, chemically defined medium for at least 18 days without any preincubation in serum-supplemented medium. Neurite outgrowth, synapse formation and spontaneous bioelectric activity, in its qualitative and quantitative aspects, were essentially the same as was observed in medium containing 20% heat-inactivated horse serum. PMID- 6272950 TI - Development of morphine-induced changes of activity in the mouse. AB - The development of spontaneous activity in response to morphine administration was studied at different postnatal ages in C57BL/6 mice. Morphine induced hyperactivity at all ages except in 3-week-old mice, in which a catatonic effect is evident. The results indicate that the neurophysiological and inhibitory systems responsible for the non-analgesic effects of morphine are not characterized by the same developmental pattern. PMID- 6272951 TI - [Study of DNA content in the lungs of rats after dust application (author's transl)]. PMID- 6272952 TI - [Results of nephroblastoma treatment in the material of the Hospital of Pediatric Surgery in Bratislava (1970-1978) (author's transl)]. PMID- 6272953 TI - [Protective action of arginine (or lysine) butyrate against the lethal effect of encelphalomyocarditis virus on mice]. PMID- 6272954 TI - [Insulin binding to isolated thyroid plasma membrane]. AB - Binding sites of insulin were detected in plasmic membranes isolated from bovine thyroid. Membrane 123I-insulin binding was inhibited by unlabelled insulin, at a lower degree by proinsulin, not at all by TSH and GH. Binding dissociation appeared to be consistent with the process of cooperative negativity. These data suggest that specific receptors binding to insulin are present in thyroid isolated plasmic membranes. PMID- 6272955 TI - [The effect of new CH-II on cAMP and cGMP levels in the cells of abdominal exudate from the mouse (author's transl)]. PMID- 6272956 TI - End-stage cytomegalic inclusions retinitis and disseminated intravascular coagulation. AB - Areas of necrosis of the retinal pigment epithelium developed in each eye of a 40 year-old renal transplant recipient. Myocardial infarctions supervened and eventually caused his death. Autopsy demonstrated cytomegalic inclusion bodies in the lungs and liver, and areas of retinal scarring and fibrin thrombi in the choriocapillaris of both eyes. The ocular features likely resulted from the resolved retinitis and preterminal disseminated intravascular coagulation. PMID- 6272958 TI - The cellular basis of metastatic bone disease in patients with lung cancer. AB - Histologic patterns of tumor-bone interaction were systematically evaluated in 80 cases of metastatic lung cancer involving bone. Patterns of tumor-bone interaction varied with the histologic type of lung cancer, reflecting the biochemical and biologic differences among the different types of lung cancer. Evidence presented here suggests that destruction of bone by metastatic lung cancer is mediated neither by direct contact of tumor cells with bone matrix nor by release of diffusible substances that lyse bone matrix. Among indirect mechanisms, the most prevalent and important was the activation of bone-lining cells by metastatic tumor. Epidermoid carcinomas in particular were associated with histologic patterns of classical bone remodelling, including osteoblastic, osteoclastic, and osteocytic activity. Adenocarcinomas showed a particularly high association with microfractures and manifested a stromal pattern consistent with release of prostaglandins. Ischemic necrosis of bone due to compression of vessels by expanding tumor mass is also a common and important mechanism. Correlation of histologic patterns with reported data on the frequency of metastases and syndromes of ectopic hormone production provides insight into the mechanism(s) of paraneoplastic hypercalcemia in patients with lung cancer. PMID- 6272957 TI - Epstein-Barr virus antibodies in patients with ataxia-telangiectasia and other immunodeficiency diseases. AB - An unusual antibody response to the Epstein-Barr virus (EBV) has been noted in patients with ataxia-telangiectasia. Of a group of 16 such patients 8 were found to have antibodies in their serum to the EBV viral capsid antigen (VCA), and 4 of them also had antibodies to the EBV early antigen (EA); antibodies to the nuclear antigen (EBNA), however, were absent in 3 of the 8. The antibody pattern persisted for more than 2 years in the patients available for follow-up study. In comparison, of 24 patients with various other immunodeficiency syndromes 9 were found to have EBV-VCA antibodies in their serum, but none of the 9 had EA antibodies and 3 lacked EBNA antibodies. Two other groups of subjects, all of whom had EBV-VCA and EBNA antibodies in their serum late after an EBV infection, were also studied; 82 had infectious mononucleosis and 55 were healthy and had no such history. EA antibodies were detected in 45 of the first group during the acute phase of the illness but persisted in only 6 of the 68 who were followed up for more than 2 years, and they were detected in only 7 of the second group.All eight lymphoblastoid cell lines established from the peripheral blood of the four patients with ataxia-telangiectasia who are still available for follow-up study express EBV-VCA, whereas most similar cell lines established from normal individuals express only EBNA. In two of these patients cell-mediated immunity, as assessed from lymphocyte transformation induced by mitogens, was markedly decreased but autologous cell-mediated immune regression of EBV-induced transformation of B (bone-marrow-derived)-lymphocytes was normal. The percentage of T (thymus-derived)-helper cells was greatly decreased in two of the three patients in whom it was measured, and the percentage of T-suppressor cells was greatly increased in one of them, but the percentage of total T-lymphocytes was within normal limits in all three.The possible significance of these findings - in particular, the persistence of EA antibodies and the diminished restriction of expression of EA - in the late development of tumours after an EBV infection in patients with ataxia-telangiectasia deserves careful attention. Finally, the apparent correlation between immunoglobulin deficiency and poor or absent EBNA antibody response warrants further study. PMID- 6272959 TI - Estrogen receptor cytochemistry in human breast cancer: status and prospects. AB - Advances in knowledge of the endocrine control of neoplastic growth can be expected to stem directly from advances in pertinent techniques. Following this theme, the present paper describes how recent development in cytochemical techniques for estrogen receptors may maintain this expectation and lead to significant progress in the basic knowledge of breast cancer that can influence the treatment approach. Cytochemical techniques have shown that the binding of estrogen to target cells and tissues can be specifically visualized: moreover, estrogen binding sites characterized cytochemically under controlled conditions have proved to fulfill the accepted criteria for estrogen receptors. Some working solutions to basic questions concerning the tumor hormone-responsiveness have been provided and more precise explanations of fundamental growth-regulatory mechanisms are expected to emerge from the approach to breast cancer by cytologic endocrinology. However, in the present state of the art and in spite of tantalizing bits of data, cytochemical techniques for estrogen receptors do not seem extensible to routine purposes, as alternative to biochemical receptor assays in selecting patients for endocrine therapy. One must have strong reservations on their clinical use because of the almost complete lack of information about correlations of cytochemical results with the response to endocrine therapy. More studies evaluating the practical validity of estrogen receptor cytochemistry are required. PMID- 6272960 TI - Cellular retinoid-binding proteins in cultured human and mouse myeloid leukemia cells. AB - Retinoic acid and retinol induced functional and morphological differentiation of human promyelocytic leukemia cells (HL-60) into mature granulocytes, but did not induce functional or morphological differentiation of mouse myeloid leukemia cells (M1). Cellular retinoic acid-binding protein, but not retinol-binding protein, was detected on HL-60 cells. Neither binding protein could be detected on M1 cells. These results suggest that retinoic acid-binding protein may be necessary for induction by retinoids of functional and morphological differentiation of myeloid leukemia cells. PMID- 6272961 TI - A short-term in vitro assay for promoter substances using human lymphoblastoid cells latently infected with Epstein-Barr virus. AB - We designed a short-term in vitro assay for detecting tumor promoters, utilizing the activation of Epstein-Barr virus (EBV) expression in EBV genome-carrying human lymphoblastoid cells. This system is composed of EBV-non-producer Raji cells as the indicator, n-butyrate as the EBV-inducer, and the test substance. After addition of the latter 2 components to the culture medium, the cells are cultivated for 48 h at 37 degrees C and the ratio of EBV early antigen (EA) expressing cells was assessed using immunofluorescence. This assay system allows for a rapid detection of the activity of the tumor promoter 12-O-tetradecanoyl phorbol-13-acetate (TPA) and its related compounds and also of the Euphorbiaceae plant extracts containing such active principles. Among several microbial products tested, teleocidin, an indole-alkaloid produced by a Streptomyces species, was also detected and had an activity level comparable to that of TPA. Other promoters, such as anthralin, phenol, Tween 60 and 80 and the carcinogenic ("initiator") substances including benzopyrene, did not react with the system. The test is simple to perform, reproducible and should be applicable for mass screening of promoter substances in the environment. PMID- 6272962 TI - Enhancement of Epstein-Barr virus-induced transformation of human lymphocytes by teleocidin. PMID- 6272963 TI - Introduction to Epstein-Barr virus and lymphoproliferative diseases in immunodeficient individuals. PMID- 6272964 TI - T-cell response to B-cells and Epstein-Barr virus antigens in infectious mononucleosis. AB - After Epstein-Barr virus (EBV) infection in vivo, B-cells with latent virus infection persist indefinitely through life. These cells grow in vitro on explanation and can be established as immortal B-cell lines. To reconcile the unlimited growth potential in vitro with the maintenance of a low proportion of B cells infected by EBV in vivo, a strict in vivo control mechanism has to be postulated. Certain aspects of this control are apparent when the primary infection is followed by infectious mononucleosis. This is characterized by lymphocytosis and the presence of activated T-cells. The T-cell proliferation is probably the manifestation of the immune response against EBV antigens. However, the reaction of T-cells upon encounter of B-blasts is also likely to contribute to the events. At present, it is difficult to detect an EBV-specific component in the action of the T-cells in the acute phase of mononucleosis exerted on B-cells. However, for the clinical course of the disease the activation of T-cells is important. The activated T-cells may control and also eliminate the B-cells infected by EBV. In addition to the immunity which develops during the disease, th immunoregulatory mechanism is likely to have a role in the inhibition of B cell proliferation. PMID- 6272965 TI - Long-term T-cell-mediated immunity to Epstein-Barr virus. PMID- 6272966 TI - Epstein-Barr virus-specific serology in immunologically compromised individuals. AB - Since B-lymphocytes are targets and a continuing habitat of Epstein-Barr virus (EBV) and the cell-mediated immune system becomes secondarily involved, one may anticipate that primary and persistent EBV infections in immunologically compromised individuals take unusual courses. Depending on the immunological defect, the clinical, hematological, and serological responses to primary EBV infections may be more or less pronounced than in immunologically competent patients. Infectious mononucleosis has per se an immunosuppressive effect which may enhance a preexisting immune defect. The persistent latent viral carrier state which regularly ensues after the primary EBV infection may become decontrolled by immunosuppressive diseases or therapy, leading rarely to illnesses referable to the virus but often to increases in the titers of antibodies to viral capsid and early antigens and/or declines in the antibody titer to EBV-associated nuclear antigen. Absence or dysfunction of different leukocyte subpopulations may account for the differential changes in antibody patterns. PMID- 6272967 TI - Documentation of Epstein-Barr virus infection in immunodeficient patients with life-threatening lymphoproliferative diseases by clinical, virological, and immunopathological studies. AB - Multiple methods, pedigree analysis, clinical evaluation, and Epstein-Barr virus (EBV)-specific serology, EBV DNA hybridization of tissues to probe for viral genome, staining of touch imprints for EBV nuclear-associated antigen, establishment of spontaneous infected B-cell lines from peripheral blood or tissues, examination of peripheral blood smears, and hematopathology studies, were used to study seven patients with the X-linked lymphoproliferative syndrome and seven additional patients with life-threatening EBV-associated diseases. These studies demonstrated EBV in the tissues of all 14 patients and immunodeficient antibody responses to EBV were documented. This virus can produce various life-threatening lymphoproliferative diseases in a variety of immunodeficient patients. PMID- 6272968 TI - Documentation of Epstein-Barr virus infection in immunodeficient patients with life-threatening lymphoproliferative diseases by Epstein-Barr virus complementary RNA/DNA and viral DNA/DNA hybridization. AB - Tissues from patients thought to have Epstein-Barr virus (EBV)-induced lymphoproliferative diseases were probed for EBV genomes using 2 independent hybridization techniques. Tissues from six patients with the X-linked lymphoproliferative syndrome, all five renal allograft recipients with immunoblastic sarcoma, and eight patients with diverse types of immunodeficiency and lymphoproliferative diseases such as fatal infectious mononucleosis or malignant lymphoma associated with antecedent immunodeficiency contained significant numbers of EBV genome equivalents per cell. The use of 2 hybridization probes is recommended to confirm the presence of EBV genomes. The finding of significant numbers of EBV genomes in tissues from patients with immunodeficiency suggests that EBV is the etiological agent of the associated lymphoproliferative diseases. PMID- 6272969 TI - Epstein-Barr virus in a malignant lymphoproliferative disorder of B-cells occurring after thymic epithelial transplantation for combined immunodeficiency. AB - A fatal disseminated polyclonal malignant lymphoproliferative disorder of B-cells (immunoblastic sarcoma) developed shortly after a second thymic epithelial peritoneal implant in a 5-yr-old girl with combined immunodeficiency. The immunodeficiency was characterized by low T-cell numbers and function, very low levels of thymic hormone, dysgammaglobulinemia, and an inability to mount a primary antibody or cell-mediated response to new antigens. At necropsy, the thymus fulfilled morphological criteria for thymic dysplasia. Epstein-Barr virus (EBV) antigen and DNA were identified in neoplastic infiltrates in the lymph nodes and thymus by immunofluorescence for the EBV nuclear antigen and by EBV specific complementary RNA/DNA hybridization. No antibodies to nuclear antigen, early antigen, or viral capsid antigen of EBV were identified in the serum. The concurrence of these events suggests that the thymic epithelial implant itself may have been instrumental in the pathogenesis of this neoplasm. It is proposed that the thymus may have provided factors which indirectly potentiated the proliferation of EBV-infected B-cells, possibly by induction of nonspecific T helper cells and perhaps through other thymic humoral factors. It is suggested that some forms of immunoblastic sarcoma, even when polyclonal, and especially those which arise in immunocompromised hosts, may, in some instances, represent an opportunistic form of EBV-induced B-cell neoplasia. PMID- 6272970 TI - Diverse familial malignant tumors and Epstein-Barr virus. AB - Continued monitoring of a family for new malignant tumors has revealed diverse immunological and neoplastic disorders during a 15-year period. In 1966, the proband developed lymphoma. In 1975, his antibody titers to Epstein-Barr virus (EBV) became elevated, and again, he developed a malignant lymphoma. He also had borderline hypo-immunoglobulin A, died of glioblastoma multiforme in 1977, and at autopsy, had adenomatous colonic polyps. His eldest brother has normal immunoglobulin levels, but developed immune thrombocytopenia in 1973 and had elevated EBV antibody titers in 1980. Another brother had hypo-immunoglobulin A, thymoma in 1965, and adenomas and adenocarcinoma of the colon. Two other brothers succumbed to glioblastoma in 1968 and 1969. The father of the proband had bronchiectasis in 1952, hypo-immunoglobulin M documented in 1972, and elevated EBV antibody titers 5 years preceding development of a malignant lymphoma. The latter contained 10 EBV genome equivalents/cell by EBV viral DNA/DNA reassociation kinetics analysis. The proband's grandmother had died of an immunoglobulin G-secreting myeloma in 1977, and his grandfather had borderline low immunoglobulin M, elevated EBV antibody titers, and hypopharyngeal carcinoma in 1980. Predisposition to oncogenesis in this family was probably inherited. PMID- 6272972 TI - Fatal Epstein-Barr virus infection in a child with acute lymphoblastic leukemia in remission. AB - A 9-year-old white boy developed a fatal primary Epstein-Barr virus (EBV) infection while receiving chemotherapy for acute lymphoblastic leukemia in remission. Histopathological findings at the height of the proliferative phase of the illness were compatible with a virally induced hemophagocytic syndrome. The infection spontaneously converted to complete aplasia of the bone marrow and lymph nodes. Serological studies disclosed that the patient had no antibodies to EBV prior to the infection, but during the acute phase he showed a spectrum and titers of antibodies to EBV-specific antigens characteristic of a current primary EBV infection. A lymph node biopsy obtained 5 weeks after onset revealed Epstein Barr nuclear antigen in approximately 50% of the cells. The boy's condition deteriorated rapidly, with disseminated candidiasis resulting in cardiorespiratory failure and death. Lymph nodes obtained at autopsy no longer contained Epstein-Barr nuclear antigen-positive cells. PMID- 6272971 TI - Clinical spectrum of lymphoproliferative disorders in renal transplant recipients and evidence for the role of Epstein-Barr virus. AB - Six renal transplant recipients with abnormal lymphoproliferative disorders were studied in an attempt to define their clinical features and the role of Epstein Barr virus (EBV) in their pathogenesis. Patients were either teenage (three) or in the sixth decade (three). The younger patients presented an average of 3 months after transplantation with fever, sore throat, and lymphadenopathy; had been markedly immunosuppressed; frequently had preceding or concomitant cytomegalovirus infections; and two of three had a rapidly fatal course. The older patients presented an average of 5 years after transplantation while on maintenance immunosuppressive drugs; in two of three cases with an oropharyngeal tumor; and had a more indolent, but frequently fatal, clinical course. The most frequent sites of biopsy-proven involvement in these patients were lymph nodes (three), the oropharynx (three), liver (three), bone marrow (three), transplanted kidney (three), colon (two), and central nervous system (two). EBV-specific antibody titers including anti-viral capsid antigen IgG, anti-viral capsid antigen IgM, anti-early antigen, and anti-Epstein-Barr nuclear antigen were serially measured in all patients. Four patients demonstrated serological evidence of a primary (one) or reactivation (three) EBV infection. No patient had significant changes in anti-early antigen or anti-Epstein-Barr nuclear antigen titers. All three patients tested for oropharyngeal shedding of EBV were positive. A touch imprint of one tumor was stained for the presence of Epstein Barr nuclear antigen, and a majority of cells were positive. EBV complementary RNA/DNA filter hybridization and/or viral DNA/DNA reassociation analysis performed on tumor biopsy specimens in five patients demonstrated multiple EBV genome equivalents per cell in all eight specimens tested. Clinical, pathological, serological, and molecular hybridization studies provide substantial evidence that EBV was the cause of these lymphoproliferative disorders occurring after renal transplantation. Impaired host defenses allow the EBV-transformed B-lymphocytes to escape normal control mechanisms. This impairment is invariable and influenced by many factors resulting in the observed spectrum of disease. Cytogenetic changes, however, may also be important. PMID- 6272973 TI - Cell-mediated immune reactions in three patients with malignant lymphoproliferative diseases in remission and abnormally high Epstein-Barr virus antibody titers. AB - Two patients with Hodgkin's disease in remission and one chronic lymphatic leukemia patient with extraordinarily high anti-Epstein-Barr virus (EBV) (viral capsid antigen) antibody titers (greater than 10,000) were selected to study a spectrum of cell-mediated immune responses, including natural killer, interferon boosted killer, antibody-dependent lymphocytotoxicity, and T-cell-mediated reactions. The purpose was to compare these reactions in patients with immunosuppression and a high EBV load who can hold their EBV-carrying cells under control with the corresponding reactions in patients with EBV-carrying lymphoproliferative disease. In contrast to the latter group, the three patients of the present study showed a less profound and less general suppression of the immune responses. Multiple effector mechanisms probably safeguard against the proliferation of EBV-transformed B-cells. Clinically manifest EBV-carrying lymphoproliferative disease occurs only in very severe immunodeficiencies effecting multiple effectors. PMID- 6272974 TI - Summary: symposium on Epstein-Barr virus-induced lymphoproliferative diseases in immunodeficient patients. PMID- 6272975 TI - Inhibition in vivo of the formation of adducts between metabolites of benzo(a)pyrene and DNA by butylated hydroxyanisole. AB - Antioxidants have been shown to inhibit the carcinogenic effects of a variety of chemical carcinogens. For example, the phenolic antioxidant butylated hydroxyanisole (BHA) has been shown to be a potent inhibitor of benzo(a)pyrene (BP)-induced neoplasia in mouse lung and forestomach. The objective of the present study was to determine whether or nt BHA, under conditions known to result in inhibition of BP-induced neoplasia, affects the formation of BP metabolite:DNA adducts. Following p.o. administration of a carcinogenic dose of [3H]BP to A/HeJ mice, radioactivity was detected in the DNA of both lung and liver. Analysis of the deoxyribonucleosides by high-pressure liquid chromatography showed that the major adduct in both tissues cochromatographed with the (+/-)-7 beta, 8 alpha-dihydroxy-9 alpha, 10 alpha-epoxy-7,8,9,10 tetrahydrobenzo(a)pyrene (BPDEI):deoxyguanosine adduct. The 7 beta, 8 alpha dihydroxy-9 beta, 10 beta-epoxy-7,8,9,10-tetrahydrobenzo(a)pyrene (BPDEII):deoxyguanosine adduct was 10 to 15% of the BPDEI adduct in both lung and liver. Another adduct, possibly derived from BP phenol(s), was also detected in lung and was 10 to 20% of the BPDEI adduct. Treatment of animals with BHA decreased the amount of the BPDEI adduct in the lung and the liver approximately 55 and 75%, respectively. The decrease in the amount of this adduct in the lung appears to correlate with the inhibition of pulmonary adenoma formation (L. W. Wattenberg, J. Natl. Cancer Inst., 50: 1541-1544, 1973; J. L. Speier, L. K. Lam, and L. W. Wattenberg, J. Natl. Cancer Inst., 60: 605-609, 1978). Thus, BHA appears to inhibit BP-induced pulmonary adenoma formation by inhibiting the amount of the BPDE:DNA adducts formed in lung. Possible mechanisms by which BHA treatment inhibits the formation of BPDE:DNA adducts are discussed. PMID- 6272976 TI - Nuclear uptake and subsequent nuclear metabolism of benzo(a)pyrene complexed to cytosolic proteins. AB - The binding of [3H]benzo(a)pyrene to proteins of rat liver cytosol, the nuclear uptake of cytosolic protein-bound [3H]-benzo(a)pyrene, and the subsequent nuclear metabolism of the polycyclic hydrocarbon were investigated. The binding of [3H]benzo(a)pyrene to cytosol had a saturable high affinity component with a Kd of 2.54 nM and a capacity of 530 fmol/mg protein. Specific binding of [3H]benzo(a)pyrene to cytosol was also assayed using sucrose density gradient analysis. Nuclear uptake of protein-bound [3H]benzo(a)pyrene was demonstrated both directly and by sucrose density gradient analysis. The nuclear benzo(a)pyrene was readily converted to metabolites which were qualitatively and quantitatively no different from nuclear metabolites of exogenously (not protein associated) added [3H]benzo(a)pyrene. PMID- 6272977 TI - Enzymatic methylation of microsomal metabolites of benzo(a)pyrene. AB - These studies suggest that the microsomal metabolism of benzo(a)pyrene (BP) produces metabolites which can be methylated by the catechol-o-methyltransferase (COMT)/S-adenosylmethionine (SAM) enzyme/donor combination. Induced microsomes converted 12 to 15% of substrate BP to polar products. Approximately 0.06% of substrate BP was recovered as COMT/SAM-reactive substances. In tests for specificity, COMT/SAM was found to react with catechols, but not with dihydrodiols, quinones, a phenol, an epoxide, or 1,4-hydroquinone. Organic extracts of COMT/[14C]SAM incubations with BP were fractionated by high performance liquid chromatography. The appearance of radiolabeled chromatographic bands required the presence of substrate BP, microsomes, and COMT/[14C]SAM. When the Ames mutagenesis assay was supplemented with COMT/SAM, a 36% reduction was observed in the number of revertant colonies induced by the microsomal oxidation of BP. In contrast, the mutagenic properties of 2-aminofluorene were not affected by COMT/SAM. These observations indicate that COMT/SAM does not generally inhibit mixed-function oxidase activity but rather reacts with substances which are activated by ring oxygenations. PMID- 6272978 TI - Identification of the mechanism of activation of 9-beta-D-arabinofuranosyladenine in human lymphoid cells using mutants deficient in nucleoside kinases. AB - The biochemical basis of cellular resistance to 9-beta-D-arabinofuranosyladenine (ara-A) and its natural purine derivative, deoxyadenosine, was investigated with two mutants of cultured human T-lymphoblastoid CCRF-CEM cells. One mutant that lacked deoxycytidine kinase activity, designated CEM/ara-C, retained about 10% of wild-type deoxyadenosine kinase and deoxyguanosine kinase activity each but maintained normal adenosine kinase or thymidine kinase activity. This suggested that in these human T-lymphoblastoid cells, as in other previously studied mammalian cells, deoxycytidine and purine deoxyribonucleosides are phosphorylated by the same enzyme. Despite this extensive reduction of purine nucleoside kinase activities, the cytotoxicity of ara-A or deoxyadenosine was not appreciably affected, decreasing by only 2.5- and 6-fold, respectively. A second mutant, isolated by selecting CEM/ara-C mutants that were resistant to ara-A, showed a 100-fold increase in resistance to ara-A cytotoxicity. This ara-A-resistant subline was deficient in the activities of two enzymes, deoxycytidine kinase and adenosine kinase, and showed a high degree of resistance to deoxyadenosine, adenosine, and pyrazofurin but not to pyrimidine analogs, such as 5 fluorodeoxyuridine or 5-fluorouridine. Further studies of ara-A and deoxyadenosine phosphorylation in wild-type and resistant cell lines disclosed that, although deoxycytidine kinase is the principal enzyme for their phosphorylation in vitro, their intracellular conversion to cytotoxic nucleotides depends on the joint action of deoxycytidine kinase and adenosine kinase rather than purine-specific deoxynucleoside kinase, as previously thought. PMID- 6272979 TI - Immunoprophylaxis of transplantable methylcholanthrene-induced murine fibrosarcomas by immunization with embryo cells expressing endogenous murine leukemia virus antigens. AB - We investigated the nature of a common tumor rejection antigen(s) in chemically induced murine fibrosarcomas. Two methylcholanthrene-induced fibrosarcomas, previously demonstrated to contain a common tumor rejection antigen(s), released infectious ecotropic murine leukemia virus and expressed the murine leukemia virus proteins, a glycoprotein with a molecular weight of 70,000 (gp70) and an envelope protein with a molecular weight of 15,000. To determine whether an antigen(s) specified by a murine leukemia virus might serve as a common tumor rejection antigen(s), primary cultures of syngeneic embryo cells or cultures of an allogeneic embryo cell line were infected with an endogenous ecotropic murine leukemia virus obtained from one of the cross-reacting fibrosarcomas; expression of infectious virus and/or viral proteins by infected and uninfected embryo cells was monitored and correlated with the results of transplantation protection tests. Uninfected allogeneic embryo cells (SC-1) did not release infectious virus or the viral protein gp70; mice immunized with SC-1 cells did not inhibit tumor growth. Uninfected syngeneic embryo cells did not release infectious virus but did release micrograms quantities of gp70 into supernatant fluids; mice immunized with uninfected syngeneic cells inhibited tumor growth in two of seven experiments. Virus-infected syngeneic and allogeneic embryo cells released both infectious ecotropic murine leukemia virus and gp70; mice immunized with virus infected cells inhibited tumor growth in 11 of 11 experiments. Growth of the two cross-reacting fibrosarcomas was inhibited in mice immunized with virus-infected embryo cells. The results indicate that antigens coded for by endogenous murine leukemia virus may function as common tumor rejection antigen on chemically induced murine fibrosarcomas. PMID- 6272980 TI - Interstitial deletion of short arm of chromosome 11 limited to Wilms' tumor cells in a patient without aniridia. AB - An interstitial deletion of the short arm of a chromosome 11 was found in Wilms' tumor cells of a patient without aniridia. The chromosomes in her peripheral blood lymphocytes were normal. The karyotype of the tumor cells is similar to that reported for the somatic cells of patients with aniridia-Wilms' tumor association. We showed that the same cytogenetic change as that seen in patients with this condition, namely, an interstitial deletion of 11p13, can occur in patients without aniridia as a mutation that is confined to tumor cells. PMID- 6272982 TI - Cell cycle-related morphological changes of feline lymphoid cells as revealed by electron microscopy. AB - The aim of this study was to correlate morphological changes in cells to known phases of the cell cycle. For this purpose, feline lymphoblastoid cells (FL74) chronically infected with feline leukemia virus were synchronized and examined by scanning and transmission electron microscopy. Scanning electron microscopy revealed that the cell surface underwent distinct changes which were found to be cell cycle specific. Morphological changes shown by transmission electron microscopy were used as ultrastructural markers to confirm the findings made by scanning electron microscopy relating to the various phases of the cell cycle. The results of these experiments suggest that the presence of morphologically mixed populations of cells in many lymphoproliferative disorders, which had been described previously, may be explained by normal cell cycle-dependent variations. PMID- 6272981 TI - Induction of the regulatory subunit of type I adenosine cyclic 3':5' monophosphate-dependent protein kinase in differentiated N-18 mouse neuroblastoma cells. AB - The expression of a adenosine cyclic 3':5'-monophosphate (cAMP)-binding protein, regulatory subunit of the type I cAMP-dependent protein kinase (Rl), and its functional significance in the differentiation of N-18 mouse neuroblastoma cells were examined. 8-Azidoadenosine cyclic 3':5'-[32P]monophosphate, a photoaffinity labeling analog of cAMP, and high-resolution sodium dodecyl sulfate polyacrylamide gel electrophoresis were used to identify and quantitate cAMP binding proteins in cell extracts. The induction of differentiation of N-18 mouse neuroblastoma cells, initiated either by adding dibutyryl adenosine cyclic 3':5' monophosphate to the growth medium or by culturing cells in medium supplemented with 1% fetal calf serum, led to a 3-fold increase in the amount of 8 azidoadenosine cyclic 3':5'-[32P]monophosphate incorporated into Rl, when assayed in vitro. This increased incorporation was attributable to an increase in the amount of Rl rather than to an increase in the affinity of Rl for 8 azidoadenosine cyclic 4':5'-[32P]monophosphate. The subunit molecular weight, isoelectric point, and immunoreactivity of Rl were found to be identical to that of the regulatory subunit of the type I cAMP-dependent protein kinase purified from bovine skeletal muscle. The increase in Rl was not accompanied by an increase in the cAMP-dependent protein kinase activity. DEAE-cellulose column chromatography confirmed the induction of Rl as a free cAMP-binding protein in the differentiated neuroblastoma cells. The possibility of a growth-dependent regulation of Rl was also examined. Addition of 2% dimethyl sulfoxide to cultures of N-18 mouse neuroblastoma cells inhibited cell growth without increasing the specific activity of Rl. Dimethyl sulfoxide had no effect on neurite outgrowth or acetylcholinesterase activity, two parameters characteristic of differentiated cells. The fact that the induction of Rl coincided with differentiation of the neuroblastoma cells suggests that the expression of Rl may be used as a biochemical index of differentiation in these cells. The presence of a free cAMP binding protein, not associated with cAMP-dependent protein kinase in neuroblastoma cells, raises important considerations concerning the action of cAMP in the regulation of growth and differentiation. PMID- 6272983 TI - Role of colostrum and milk in the natural transmission of the bovine leukemia virus. AB - The role of colostrum and milk in the transmission of the bovine leukemia virus (BLV) was examined by monitoring the development of BLV infection in calves that were fed since birth on colostrum and milk from their BLV-positive dams and then reared in complete or partial isolation from infected cattle. Twenty-one of the 25 calves raised in complete isolation remained negative for BLV until the last evaluation. At this time, 14 calves were older than 28 months of age, and seven were 16 to 20 months old. Three calves in this group became BLV positive before the age of five months, and one became so at the age of 12 months. Of the 16 calves raised in partial isolation, two were positive at the ages of 11 and 18 months, respectively. The other 14 calves remained negative during the 26 to 29 months of observation. It is not known if the six animals that became BLV positive in these two groups were infected by milk, or prenatally, or during contact with their infected dams. While only six of the 41 calves raised in isolation became infected, all 18 calves raised in contact with BLV-positive cows became infected before the age of 26 to 29 months of age, and 12 were positive at 16 to 20 months of age. Thus, it is apparent that, under natural conditions, milk borne transmission of BLV, if it occurs at all, is much less frequent than contact transmission, despite the fact that, as shown in previous studies, infectious BLV is present in the colostrum and milk of most BLV-positive cows. Passively acquired maternal antibodies to BLV were probably responsible for the resistance of the calves to milk-borne infection. PMID- 6272984 TI - In vitro metabolism of acrylonitrile to 2-cyanoethylene oxide, reaction with glutathione, and irreversible binding to proteins and nucleic acids. PMID- 6272985 TI - Glucocorticoid-stimulated increase in chemotactic peptide receptors on differentiating human myeloid leukemia (HL-60) cells. AB - The effect of dexamethasone on the HL-60 human promyelocytic leukemia cell line was studied using a radioligand-binding assay for a chemotactic peptide receptor that is found on the surface of mature polymorphonuclear leukocytes. When dimethyl sulfoxide (DMSO) is added to cultures, HL-60 cells undergo morphological and functional differentiation over a period of 6 to 7 days. Differentiation is accompanied by increased binding of the synthetic N-formylated chemotactic peptide N-formylmethionylleucyl[3H]phenylalanine. Binding of N formylmethionylleucyl[3H]phenylalanine to cells induced to differentiate with DMSO was temperature dependent, specific, saturable, of high affinity, reversible, and proportional to cell number. Dexamethasone increased the number of N-formylated chemotactic peptide receptors in cultures of differentiating HL 60 cells without affecting the affinity of the receptors for the tritiated peptide. This steroid response was dose dependent, proportional in magnitude to glucocorticoid activity, and abolished by cycloheximide. Dexamethasone had little effect on N-formylmethionylleucyl[3H]phenylalanine binding unless DMSO was also present in culture or the cells were first induced by DMSO to differentiate. These results suggest that the glucocorticoid stimulated increase in number of N formylated chemotactic peptide receptors was mediated by the high-affinity glucocorticoid receptor, involved protein synthesis, and was apparently dependent on differentiation. PMID- 6272986 TI - Relationship between biological responsiveness to phorbol esters and receptor levels in GH4C1 rat pituitary cells. AB - During a 24-hr incubation of GH4C1 cells with phorbol esters or thyrotropin releasing hormone, there is a decrease in the number of phorbol esters receptors (down-modulation). The purpose of this study was to investigate whether this decrease in receptor number attenuated cellular responsiveness to subsequent challenge with phorbol esters. Accordingly, cellular sensitivity to a phorbol ester-mediated biological response, namely the decrease in binding of epidermal growth factor, was compared in control and down-modulated cells. This phorbol ester-mediated event is closely associated with phorbol ester receptor occupancy, and it is therefore an effect for which altered dose-response characteristics correlating with alterations in the number of phorbol ester receptors could be anticipated. In fact, during a 48-hr exposure to phorbol esters, GH4C1 cells become refractory to the effect on epidermal growth factor binding. This time course is similar to that for the loss of phorbol ester receptors. However, when cells are down modulated by pretreatment with phorbol ester or thyrotropin releasing hormone and then (re)challenged with phorbol ester, no differences in dose-response characteristics were observed between control and down-modulated cells. We therefore conclude that phorbol ester receptor down-modulation does not affect cellular responsiveness to phorbol esters, at least when decreased epidermal growth factor binding is used as the marker for the phorbol ester mediated event. PMID- 6272987 TI - Repair of psoralen-treated DNA by genetic recombination in human cells infected with herpes simplex virus. AB - Herpes simplex virus type 1 was treated with 4,5'-8-trimethylpsoralen (psoralen) plus near-ultraviolet light in order to produce lesions (monoadducts and DNA cross-links) in the viral DNA. Human fibroblasts were infected by damaged virus under conditions in which either a single virus particle or several particles entered a given cell, and the fraction of virus-producing cells was determined. This fraction was significantly greater for multiply infected cells than for singly infected cells, indicating that the psoralen lesions are repaired more efficiently in the present of homologous, damaged DNA (multiplicity reactivation). Evidence is presented that herpes simplex virus may code for functions which participate in its own repair, both during multiplicity reactivation and during repair which occurs in singly infected cells: (a) host cells deficient in repair of lesions induced by psoralen (xeroderma pigmentosum) or the DNA cross-linking agent mitomycin C (Fanconi's anemia) exhibited normal levels of multiplicity reactivation of psoralen-treated herpes virus; (b) while xeroderma pigmentosum cells have been previously shown to be deficient in repair of psoralen-treated adenovirus under conditions of single infection, herpes virus is repaired at near normal levels in these same cells. Recombination levels between genetically marked pairs of herpes viruses were found to increase after treatment of the parental viruses with psoralen, suggesting that psoralen damage stimulates genetic recombination. This stimulation provides convincing evidence for a repair pathway in which genetic recombination between damaged viral genomes can lead to the production of viable virus. PMID- 6272988 TI - Divalent cation-independent aggregation of rat-1 fibroblasts infected with a temperature-sensitive mutant of Rous sarcoma virus. AB - Rat-1 fibroblasts infected with the temperature-sensitive transformation mutant LA 24 of Rous sarcoma virus have a high rate of divalent cation-independent homotypic cell aggregation when grown at the permissive temperature, 34 degrees. Cells grown at the nonpermissive temperature, 39 degrees, have a low rate of homotypic cell aggregation. Hyaluronic acid is involved in the homotypic aggregation of permissively grown cells since aggregation is blocked by either treatment of the cells with hyaluronidase or the presence of exogenously added hyaluronic acid. Despite their low rate of homotypic aggregation, nonpermissively grown cell are capable of heterotypically adhering to premissively grown cells. The formation of heterotypic aggregates is blocked by treatment of the nonpermissively grown cells with hyaluronidase but not by treatment of permissively grown cells with hyaluronidase. These results provide evidence that the divalent cation-independent aggregation of Rat-1 LA 24 cells is mediated by interactions between hyaluronic acid and other adhesive components on the cell surface. PMID- 6272989 TI - Frequency of 1,25-dihydroxyvitamin D3 receptor in human breast cancer. AB - Receptors for 1,25-dihydroxyvitamin D3 have been shown to exist in cultured breast cancer cells and in primary breast cancers. It is reported here that 1,25 dihydroxyvitamin D3 receptor (1,25-DR) was present in 80% of 54 unselected breast cancers. The concentration of 1,25-DR in the 43 receptor-positive tumors was 1.9 +/- 0.4 fmol/mg protein (S.E.). There was no correlation between 1,25-DR presence or concentration and the age of the patient or the concentration of estrogen, progesterone, androgen, or glucocorticoid receptors. 1,25-DR was also found in two of three renal cortical carcinomas but only in three of 14 gastrointestinal tract carcinomas. The relatively low concentration of 1,25-DR in these breast cancers, compared with that found in cultured breast cancer cells, is partially explained by incomplete "exchange" with occupied receptors. Since the serum vitamin D-binding protein is not precipitated from serum itself or from tissue homogenates using the polyethylene glycol method, artifactual 1,25-DR levels due to the inevitable contamination of tissue specimens with this protein can be excluded. These findings indicate that 1,25-DR is not a nonspecific marker of cancer. The high frequency of 1,25-DR in the breast cancers may be related to the calcium-transporting ability of breast cancer cells which allows them to grow as osteolytic metastases. PMID- 6272990 TI - G1-phase arrest of cultured human leukemic T-cells induced by deoxyadenosine. AB - Cultured human T-cell leukemia lymphocytes have enhanced sensitivity to growth inhibition by deoxyadenosine. We have used flow cytometry to investigate the mechanism of deoxyadenosine toxicity in cultured T-leukemic cells. Comparative studies on deoxyadenosine-resistant Epstein-Barr virus-transformed B-lymphocyte cell lines were also performed. After exposure of T-cells to low concentrations of deoxyadenosine (3 microM), in the presence of an adenosine deaminase inhibitor (erythro-9-[3-(2-hydroxynonyl)]adenosine), accumulation of cells of cells with a G1 DNA content was demonstrated. In contrast, B-cell lines showed a similar degree of growth inhibition after exposure to 200 to 400 microM deoxyadenosine but were blocked in S phase. The T-cell G1 block was associated with a rise in the deoxyadenosine triphosphate pool, and both these phenomena were prevented by the addition of deoxycytidine. The biochemical mechanism of this G1 block induced by deoxyadenosine in T-cells is not understood. PMID- 6272991 TI - Response to doxorubicin and mitomycin in cholangiocarcinoma: a case report. PMID- 6272992 TI - Evaluation of the lipid-soluble diaminopyrimidines, metoprine and etoprine, in the avian sarcoma virus rat glioma model. AB - Using the avian sarcoma virus (ASV) rat glioma model, we have evaluated the in vivo effectiveness of two lipid-soluble folate antagonists, metoprine (DDMP) and its 6-ethyl analog etoprine. When adult Fischer 344 rats, which were previously inoculated with ASV as newborns, received DDMP ip, high levels of this drug accumulated in both normal and gliomatous brain tissue, as well as in lung and liver. When animals bearing ASV-induced gliomas were treated with DDMP or etoprine with or without citrovorum factor (calcium leucovorin), there was no increase in survival, even though the drugs were administered at maximally tolerated doses. PMID- 6272993 TI - An update on the biochemistry of 5-fluorouracil. PMID- 6272994 TI - Cefotaxime: clinical and bacteriologic evaluation. AB - Cefotaxime, a new parenteral beta-lactamase-resistant cephalosporin, was tested against 78 cephalothin-resistant clinical isolates of Enterobacteriaceae and Pseudomonas species. The anti-bacterial activity of the drug was compared with activities of cefazolin and cefuroxime and was found considerably more active. Most of the cephalothin-resistant strains were resistant to cephazin, too. The antipseudomonal activity of cefotaxime was found limited. The drug was found effective against 23 infections in hospitalized patients with only one clinical failure and two colonizations with cefotaxime-resistant bacteria. Cephalosporinases hydrolyzing cefotaxime were detected in two cefotaxime resistant strains. Also, beta-lactamases hydrolyzing cefoxitin and cefuroxime were produced by the cefotaxime-resistant isolates studied. PMID- 6272995 TI - Subhypothalamic grafts of human pituitary adenomas in total-body irradiated rats. AB - Human pituitary adenomas proliferate neither in cell culture nor in athymic "nude" mice. We propose that one or several of the humoral factors necessary for the growth of pituitary adenomas is missing in these experimental environments. The purpose of our experiments was to examine the possible influence of the hypothalamus in supporting cellular proliferation, and thus adenoma growth. Fragments from four human pituitary adenomas (three pituitary prolactinomas; one ACTH-secreting adenoma) were transplanted into the pituitary fossa of total-body irradiated, hypophysectomized rats. The rats were killed after two weeks and perfused with a mixture of formalin and India ink. Histologic examination of serial sagittal sections through the pituitary fossa and the adjacent brain showed: vascularization of the grafts from the pituitary stalk and from the scar tissue in the sphenoid bone; survival of some adenomas; and numerous mitoses in an ACTH-secreting specimen obtained from a patient who had Cushing's disease. We conclude from these experiments that as yet unidentified hypothalamus factors are essential for the growth of certain types of pituitary adenomas. PMID- 6272996 TI - In vitro sensitization with HSV-infected cells induces T micro cells to become cytotoxic T gamma cells which are theophylline resistant. PMID- 6272997 TI - Relationship between production and release of lymphocyte-activating factor (interleukin 1) by murine macrophages. 1. Effects of various agents. PMID- 6272998 TI - [About the possibility of fallopian tubes patency evaluations by means of 99mTc pertechnetate and comparison of radiation loads due to radioisotopic and x-ray examinations (author's transl)]. PMID- 6272999 TI - [Vagal chemodectoma]. PMID- 6273000 TI - 2-(Alkylthio)penem-3-carboxylic acids. III. Synthesis of 6-ethylpenems. PMID- 6273001 TI - [Application of anticomplement immunoenzymatic method for the detection of EBNA in carcinoma cells and normal epithelial cells from nasopharynx (author's transl)]. PMID- 6273002 TI - [Effects of "Beijing Composite Recipe No. 2 for coronary disease" on cAMP content of washed rabbit platelets (author's transl)]. PMID- 6273003 TI - [Antibodies to EB viral capsid antigen in rhesus monkey sera (author's transl)]. PMID- 6273004 TI - [Studies on an improved technic to increase the positive detection rate of rotavirus under electron microscope (author's transl)]. PMID- 6273005 TI - Hormone receptors in renal cell carcinoma. Their utility as predictors of response to endocrine therapy. AB - We have performed 23 estrogen and 24 progesterone receptor assays on tumor specimens from 26 patients with renal cancer, ten of whom subsequently received hormonal therapy for metastatic disease. Only one of these specimens contained measurable estrogen receptor levels, three contained low, but measurable progesterone receptor levels, and the remaining specimens contained no measurable estrogen or progesterone receptors. None of ten patients with metastases showed evidence of an objective response to hormonal therapy. Our data suggest that few, if any, renal cancers have high titers of estrogen or progesterone receptors, and that those patients whose tumors have low receptor titers are unlikely to respond to hormonal therapy. PMID- 6273006 TI - A phase II clinical study of mAMSA in small cell carcinoma of the lung. AB - Thirteen patients with small cell carcinoma of the bronchus that had become resistant to conventional chemotherapy were given 4'-(9 acridinylamino)methanesulphon-m-anisidide (mAMSA) at a dose of 90 or 120 mg/m2 at 3-week intervals. Twenty percent of the courses at 90 mg/m2 produced marked myelosuppression. No responses were observed. Median survival from the start of treatment with mAMSA was 5 weeks. PMID- 6273008 TI - Influence of the location of left anterior descending coronary artery stenosis on left ventricular function during exercise. PMID- 6273007 TI - Studies on phosphorylation of canine cardiac sarcoplasmic reticulum by calmodulin dependent protein kinase. AB - Two endogenous protein kinase activities, cAMP-dependent and calmodulin-Ca2+ dependent, are associated with isolated cardiac sarcoplasmic reticulum (SR) vesicles. Both kinases phosphorylate an endogenous substrate of approximately 22,000 daltons (phospholamban). The phosphorylation of phospholamban by either the intrinsic or by exogenous cAMP-dependent protein kinase is found to be Ca2+ independent between 0.05 and 100 microM free Ca2+. Calmodulin-dependent phosphorylation, on the other hand, does not require cAMP and is absolutely dependent on the presence of free Ca2+ over a concentration range that corresponds to physiological levels (10(-7) to 10(-5) M). Phosphorylation of SR vesicles by both kinases is additive and the extent of saturation of the cAMP specific sites has no effect on the degree of stimulation by calmodulin or its Ca2+-dependence. Trifluoperazine, an inhibitor of calmodulin, inhibits calmodulin dependent phosphorylation without affecting cAMP-dependent phosphorylation, indicating the presence of two types of kinases. This is made further evident by the selectivity of each kinase for exogenous substrates. Whereas cAMP-dependent protein kinase appears to phosphorylate histone ILA (a basic protein) preferentially, calmodulin-dependent protein kinase prefers phosvitin (an acidic protein). PMID- 6273009 TI - Response of the systemic and pulmonary circulation to converting-enzyme inhibition (captopril) at rest and during exercise in hypertensive patients. AB - Twenty sodium-replete patients with hypertension were allocated either to a placebo or to a captopril treatment group. Each patient was investigated in rest recumbent (RR) and rest-sitting (RS) positions and during an uninterrupted, graded, submaximal exercise test (up to the anaerobic threshold) before treatment, and with a similar protocol 75 minutes after treatment with captopril or placebo on the same morning. Captopril decreased brachial intraarterial pressure by 7/4 mm Hg at RR, by 16/10 mm Hg at RS, and by 19/10 mm Hg during exercise (p less than 0.001), based on a decrease of systemic vascular resistance (p less than 0.001). Slight increases of cardiac output and of heart rate were noted at rest; cardiac output was not significantly affected during exercise, but the increase of heart rate of 2.4 beats/min was significant (p less than 0.01). Captopril decreased pulmonary artery (p less than 0.05) and capillary wedge pressures (p less than 0.001), with unchanged pulmonary vascular resistance. The data indicate that the action of captopril is characterized by arteriolar and possibly venous dilatation both at rest and during exercise. Pulmonary vascular resistance, however, is not affected. PMID- 6273010 TI - Limitation of experimental infarct size by an angiotensin-converting enzyme inhibitor. PMID- 6273011 TI - Measurement of absolute left ventricular volume from gated blood pool studies. PMID- 6273013 TI - Optimized assay for serum angiotensin-converting enzyme activity. AB - This optimized two-point kinetic assay for serum angiotensin-converting enzyme is based on the colorimetric determination of hippurate with cyanuric chloride/dioxan reagent. The hippurate is released from hippuryl-L-histidyl-L leucine by angiotensin-converting enzyme in the presence of chloride ion. CVs for the method (within-run and between-run) ranged from 2.1 to 3.2%. Linearity extends up to 200 U/L. Results are unaffected by lipemia and icterus. Hemoglobin in concentrations greater than 1.5 g/L causes a slight negative interference. Reference intervals for men and women are 22-82 U/L and 25-69 U/L, respectively. PMID- 6273012 TI - Ion channels in membranes. AB - The membranes that separate the myocardial cell interior from the extracellular space and delimit compartments within the myocardial cell represent ion impermeable phospholipid barriers. Embedded in these phospholipid membranes are intrinsic membrane proteins, some of which serve as ion channels. The voltage sensitive ion channels that control the sarcolemmal action potential appear to be highly regulated intrinsic membrane proteins that contain "gates" that respond to changing membrane potential by opening and closing an ion-selective "pore" that allows specific ions to cross the membrane. Pharmacologic blockade of the sarcolemmal ion channels is selective, not only for individual classes of ion channels, but also for specific states of a given type of channel. The basis for this selectivity remains unclear, but may derive from a preferential interaction between a given drug and a specific type of ion-channel protein, or a selective drug action on a structurally specific region of the membrane phospholipid that is in intimate contact with the ion-channel protein. PMID- 6273015 TI - Influence of bilirubin on determination of angiotensin-converting enzyme. PMID- 6273014 TI - Unexplained increase in serum creatine kinase isoenzyme MB activity in a lung cancer patient. AB - In this case of mixed small cell--large cell cancer of the lung in an elderly woman, creatine kinase (EC 2.7.3.2) isoenzymes were assayed serially because of chest pain. The proportions of serum CK-BB and CK-MB isoenzyme activities were persistently above normal (CK-MB 10-18%, normal less than 5%). Electrocardiograms revealed no signs of ischemia or infarction. At autopsy no gross or microscopic infarction or inflammation of the heart was seen. There was also no infarction of smooth or skeletal muscle. The tumor was the probable source of most of the circulating CK-MB isoenzyme. Future cases may pose a similar diagnostic dilemma: differentiating creatine kinase that is present as a result of myocardial infarction from tumor-related CK-MB. Whether or not CK-MB assay could be useful in detecting tumors remains to be investigated. PMID- 6273016 TI - Human serum inhibitors of collagenase as revealed by preparative isoelectric focusing. AB - Single step separation of pooled normal human serum by means of preparative isoelectric focusing in the range from pH 3.5--9.8 revealed more heterogeneous inhibition of collagenolytic activity than previously reported. Essentially three inhibition zones were resolved. According to their electrophoretic behaviour the respective serum fractions displaying inhibitory activity were designated alpha-, beta- and gamma-collagenase inhibitors. The main component responsible for collagenase inhibition in the alpha-zone was found to be alpha 2-macroglobulin. In the beta-zone inhibitory activity focused around pH 6.3. In the gamma-range a non-dialysable cationic component focusing at pH 9.2 was also able to decrease collagenolytic activity derived from rheumatoid arthritis synovial culture supernatant. These findings were supported by single step separation of serum on DEAE-anion exchange chromatography. PMID- 6273017 TI - Direct assay of cortisol in human saliva by solid phase radioimmunoassay and its clinical applications. AB - Direct determination of cortisol (F) in human saliva and its clinical applications were investigated. For this purpose, a solid phase radioimmunoassay (RIA) was carried out. The detection limits for F in 50 and 400 microliter of saliva were 0.125 and 0.0156 micrograms/100 ml, respectively, and the intra-assay coefficient of variation was 5.9--12.2%. Salivary F levels measured by this method were in good agreement with those by the RIA after extraction with dichlormethane reported previously. Salivary F levels were studied in 10 healthy adults with regard to diurnal rhythms, in rapid ACTH tests, and following intravenous injections of 20 mg F, and they showed almost similar changes to serum unbound F levels. Diurnal variations of salivary F in 8 normal children were nearly the same as those in healthy adults. In patients with Cushing's syndrome, neither normal diurnal rhythm of salivary F nor suppressed salivary F with dexamethasone were observed. Patients with adrenocortical insufficiency showed a lack of responsiveness in salivary F levels to stimulation with ACTH or lysin-vasopressin. These findings suggest that salivary F can be measured directly by the solid phase RIA and accurately reflects serum unbound F. The method is simple, accurate and useful for assessing adrenocortical function, especially in pediatric subjects and/or outpatients. PMID- 6273018 TI - Autonomic neural control mechanisms of substrate and hormonal responses to acute hypoglycaemia in man. AB - The contributions of adrenergic and cholinergic mechanisms to recovery from acute hypoglycaemia induced by insulin (0.15 units/kg i.v.) were examined in eleven normal subjects, six subjects with a pre-ganglionic sympathectomy (adrenergic denervation) and six sympathectomized subjects given atropine (combined adrenergic denervation and cholinergic blockade). Blood glucose recovery was impaired only in the sympathectomized subjects given atropine. The blood lactate response was reduced and the rise in free fatty acids was delayed in both groups of sympathectomized subjects, in whom the normal rises of plasma cyclic AMP and noradrenaline were absent. The plasma pancreatic glucagon response was appropriate to the prevailing blood glucose concentrations in all three groups. The cortisol response was impaired and the pattern of ACTH secretion was abnormal in sympathectomized subjects given atropine. Growth hormone levels were higher in both sympathectomized groups. Blood glucose homeostasis was impaired during combined adrenergic denervation and cholinergic blockade. Glucagon secretion was activated independently of vagal control. In the sympathectomized group given atropine, the rise in plasma cortisol was blunted despite a greater degree of hypoglycaemia. A blockade of central cholinergic receptors producing impaired activation of ACTH secretion at hypothalamic level may explain, at least in part, this delayed restoration of normoglycaemia. PMID- 6273019 TI - Metabolic response to subtotal thyroidectomy in propranolol-treated thyrotoxic patients. AB - Changes in plasma cortisol, glucose, cyclic AMP, blood leucocytes, pulse rate and body temperature were measured during and for 24 h after subtotal thyroidectomy in eight propranolol-treated (120-160 mg per day) mild to moderate thyrotoxic patients and eight patients with a non-toxic goitre. The results showed a similar metabolic response to surgery in the two groups without any trends towards a hypermetabolic response in the toxic group. The accumulating evidence of the safety of preoperative preparation with propranolol in patients with mild to moderate thyrotoxicosis is thus supported by our results. PMID- 6273020 TI - Patterns of plasma cortisol and ACTH concentrations in patients with Addison's disease treated with conventional corticosteroid replacement. AB - Plasma cortisol and adrenocorticotrophin hormone (ACTH) profiles were estimated in twelve patients with Addison's disease following randomized oral administration of either cortisone acetate (25 mg) or hydrocortisone (20 mg) alternately, at 0900 h on consecutive days. Normal corticosteroid replacement therapy was discontinued from 1200 h on the day prior to the study period. In four patients elevated basal plasma ACTH concentrations were not suppressed to the limit of detection following the administration of either drug, and in three of these no suppression was found following the prolonged administration of pharmacological doses of dexamethasone. Diminished sensitivity of pituitary ACTH secretion to cortisol inhibition may result from chronic loss of negative feedback before and/or after diagnosis and treatment. In three patients elevated basal plasma ACTH concentrations were suppressed adequately during the administration of either drug, but in five, low basal ACTH concentrations following corticosteroid withdrawal suggested chronic inhibition of anterior pituitary corticotrophs by over-replacement with glucocorticoid. However, further study is necessary to determine whether the estimation of ACTH profiles is a more accurate reflection of the adequacy of corticosteroid replacement than the estimation of cortisol profiles alone, and whether this estimation leads to an improvement in patient management. Hydrocortisone (20 mg) achieved higher mean cortisol levels and lower mean ACTH levels than cortisone acetate (25 mg), but either drug may be suitable for glucocorticoid replacement provided the dose is tailored to the individual needs. PMID- 6273021 TI - The effects of a met-enkephalin analogue on ACTH, beta-LPH, beta-endorphin and MET-enkephalin in patients with adrenocortical disease. AB - Thirteen patients with either Addison's disease, or Cushing's disease treated by bilateral adrenalectomy, were infused with the long-acting met-enkephalin analogue DAMME. In patients with Addison's disease significant and pronounced falls in ACTH and N- and C-terminal beta-LPH were seen; chromatography suggested that beta-endorphin fell concomitantly. Three out of four patients with Cushing's disease who had not received pituitary irradiation, also showed a decrease in plasma ACTH and N- and C-terminal beta-LPH; however, no change was seen in any of the irradiated patients. The changes were naloxone reversible. The levels of plasma met-enkephalin were normal and did not change after DAMME in any group of patients. These results are interpreted as suggesting that there are inhibitory opiate receptors controlling the release of ACTH, beta-LPH, and beta-endorphin. PMID- 6273022 TI - Prolonged remission in florid Cushing's syndrome following metyrapone treatment. AB - Two patients presenting with diabetes mellitus and hypokalaemia resulting from markedly increased ACTH and cortisol secretion are described. Neither patient showed any evidence of a tumour and both responded dramatically to treatment with metyrapone in that all abnormal clinical features disappeared, ACTH concentrations returned to normal and both patients showed prolonged remission after metyrapone treatment was stopped. One patient relapsed after a severe viral illness and the administration of dexamethasone and cortisone. It is suggested that these cases may represent an unusual form of Cushing's syndrome in which ACTH secretion is stimulated by increasing concentrations of cortisol. When these are reduced by metyrapone administration ACTH secretion falls in parallel and prolonged remission of disease may result. PMID- 6273023 TI - Parathyroid function after parathyroidectomy: evaluation by measurement of urinary cAMP. PMID- 6273025 TI - Effects of valproic acid, naloxone and hydrocortisone in Nelson's syndrome and cushing's disease. AB - Two patients with Nelson's syndrome and one patient after bilateral adrenalectomy for Cushing's disease, without any evidence of Nelson's syndrome, were studied with respect to the effect of hydrocortisone, naloxone and valproic acid (a GABA transaminase inhibitor) on ACTH secretion. Hydrocortisone suppressed plasma ACTH concentrations to normal in the patient without Nelson's syndrome, but failed to do so in the two patients with Nelson's syndrome. Naloxone and valproic acid caused a decline in plasma ACTH concentrations in the patients with Nelson's syndrome, but produced no change in the patient without Nelson's syndrome. Secretion of ACTH may thus be influenced by both opiate peptide and by gamma aminobutyric acid, as well as by the cortisol concentration, these agents may act at different sites to inhibit ACTH release by the tumour. PMID- 6273024 TI - The short metyrapone test: comparison of the plasma ACTH response to metyrapone with the cortisol response to insulin-induced hypoglycaemia in patients with pituitary disease. AB - After a single dose of 2 g metyrapone given with a snack at 2300 h, blood was taken for estimation of plasma adrenocorticotrophin hormone (ACTH) concentrations at 0800 h the next morning from normal individuals and from patients with pituitary disease. Plasma ACTH concentrations following metyrapone were significantly lower in patients with pituitary disease with no evidence of anterior pituitary hormone deficiency (mean 90.0 +/- SD 63.9 ng/l) when compared with normal individuals (mean 182.5 +/- SD 14.1 ng/l, P leads than 0.01) but significantly greater than in patients with pituitary disease in whom a deficiency of one or more anterior pituitary hormones was demonstrated (mean 50.8 +/- SD 14.1 ng/l, P less than 0.01). It is concluded that the short metyrapone test is a sensitive method of detecting minor degrees of impairment of the negative feedback control of aCTH secretion that would be unrecognized by conventional assessment by insulin-induced hypoglycaemia. The clinical relevance of impaired ACTH release in response to a single dose of metyrapone in patients with pituitary tumours, in whom pituitary function is otherwise shown to be adequate, needs to be determined. PMID- 6273026 TI - Development of a radioimmunoassay for an amino-terminal peptide of pro-opiocortin containing the gamma-MSH region: measurement and characterization in human plasma. AB - A radioimmunoassay has been developed for the N-terminal region of human pituitary pro-opiocortin (N-POC), the common precursor protein of ACTH and beta LPH, using an antiserum which recognizes residues near the gamma-MSH region. The concentrations of greater than 300 ng/l of immunoreactive peptide were determined in unextracted human plasma, the relative molecular mass of the reacting fragments corresponding to a seventy-seven amino acid glycoprotein. The concentrations of immunoreactive N-POC peptides were correlated with those of ACTH in plasma obtained from patients with various disorders of the hypothalamic pituitary-adrenal axis. PMID- 6273027 TI - Is 1,25-dihydroxy-cholecalciferol harmful to renal function in patients with chronic renal failure? AB - Seventeen undialysed adult patients with chronic renal failure took part in a controlled study of the effects of 1,25(OH)2D3 and D3. After a 6-month observation period the patients were allocated at random to two groups for 6 months of treatment with either 1,25(OH)2D3 (mean dose 0.5 microgram daily) or D3 (dose 100 microgram daily). Treatment was then discontinued and the patients were studied for a further 6 months. Serum iPTH was decreased in both groups but most markedly in the 1,25(OH)2D3 group in which the iPTH values became normal. Serum creatinine increased during treatment in both groups. In the group receiving 1,25(OH)2D3 this was coupled to an increase in serum calcium within the normal range. Our data demonstrate that 1,25(OH)2D3 treatment in patients with chronic renal failure leads to a further reduction in renal function, which may be partially reversible. Physicians should therefore be reluctant to give vitamin D analogues to patients with chronic renal failure unless they have severe symptomatic renal osteodystrophy. PMID- 6273028 TI - A possible direct effect of 24,25-dihydroxycholecalciferol on the parathyroid gland in patients with chronic renal failure. AB - Seventeen undialysed adult patients with chronic renal failure took part in a controlled study of the effects of 1,25(OH)2D3 and D3. After a 6-month observation period the patients were allocated at random to two groups for 6 months of treatment with either 1,25(OH)2D3 (mean dose 0.5 microgram daily) or D3 (dose 100 microgram daily). The treatment was then discontinued and the patients were studied for a further 3 months. In the 1,25(OH)2D3 group the mean serum concentration of 1,25(OH)2D rose significantly during treatment, whereas serum concentratins of 25OHD and 24,25(OH)2D remained unchanged. In the D3 group there was a highly significant increase in serum concentrations of 25OHD and 24,25(OH)2D, whereas serum 1,25(OH)2D remained unchanged. There was a significant fall in serum iPTH in both treatment groups. This fall was unrelated to serum calcium in the D3 group unlike the findings in the 1,25(OH)2D3 group. The data support previous experimental evidence that serum iPTH can be suppressed by 24,25(OH)2D3 and suggest that this analogue may be of clinical importance in the treatment of chronic renal failure without inducing hypercalcaemia. PMID- 6273029 TI - Studies on circulating met-enkephalin and beta-endorphin: normal subjects and patients with renal and adrenal disease. PMID- 6273030 TI - Genetics of the low density lipoprotein receptor: II. Genetic control of variation in cell membrane low density lipoprotein receptor activity in cultured fibroblasts. AB - Fibroblast low density lipoprotein (LDL) plasma membrane receptor activity, measured as 125I-LDL association (plasma membrane binding plus intracellular accumulation) and degradation was determined in cell strains from 14 monozygotic (MZ) and 21 like-sexed dizygotic (DZ) normolipidemic twin pairs. The twins were between 57 and 62 years old and had liver apart for an average of 38 years (range 0-60). The intrapair differences were significantly smaller in MZ than in DZ twin pairs in fibroblast 125I-LDL association as well as degradation assays (P less than 0.05). These findings suggest a genetic influence on normal variation in LDL receptor activity in vitro. In two MZ pairs discordant for psoriasis, the psoriatic twin had markedly lower LDL receptor activity than the cotwin. PMID- 6273031 TI - Genetics of the low density lipoprotein receptor: III. Evidence for multiple normal alleles at the low density lipoprotein receptor locus. AB - Fibroblast association (plasma membrane binding plus intracellular accumulation) and degradation of radioiodinated low density lipoprotein (125I-LDL) index plasma membrane LDL receptor activity. Cultured fibroblasts from 23 subjects affected with familial hypercholesterolemia (HC) and from 95 subjects without HC (non-HCs) were tested for 125I-LDL association and degradation. Both LDL receptor activity indices were twice as high in non-HC and HC heterozygous cell strains. This is compatible with a major gene effect on LDL receptor activity. However, a considerable overlap between non-HC and HC heterozygous values was found in the 125I-LDL association assay [median (range) 970 (330-2500), and 450 (250-490), respectively] and in the degradation assay [median (range) 810 (280-2020), and 470 (160-790), respectively]. The values are expressed as ng 125I-LDL X mg cell protein-1 X 4.5 h-1. These great overlaps in the LDL receptor activity indices support the view that the influence of LDL receptor activity on the HC phenotype may be smaller than believed previously. Furthermore, for the diagnosis of HC, these LDL receptor activity assays are far more expensive and have less sensitivity and specificity than simple serum cholesterol determination. The LDL receptor-dependent 125I-LDL association values for the HC heterozygous individuals clustered into four groups. Family data supported the hypothesis that this variation could be due to four different LDL receptor variants, each coded for by different alleles at the LDL receptor locus. If confirmed, this finding may have implications for the understanding of the variable expression of HC and also of the genetic impact on lipoprotein metabolism and susceptibility to atherosclerosis in non-HCs. PMID- 6273032 TI - Genetics of the low density lipoprotein receptor: I. Low density lipoprotein receptor activity in cultured fibroblasts from subjects with or without familial hypercholesterolemia. AB - Six indices of low density lipoprotein (LDL) receptor activity were assayed in cultured fibroblasts from seven subjects with familial hypercholesterolemia (HC) and six subjects without HC (non-HCs). Four non-HCs, three HC heterozygotes and one HC homozygous proband belonged to one kindred (kindred A). The proband's fibroblast 125I-LDL processing values fell within or were slightly above the range defined by fibroblasts from three "receptor-negative" HC homozygotes. Thus, the plasma membrane receptor defect in this kindred is probably of the "receptor negative" category. LDL receptor-dependent 125I-LDL processing was about twice as high in fibroblasts from non-HCs as in those from HC heterozygotes belonging to kindred A. The segregation pattern of LDL receptor activity in this kindred was compatible with control by a single gene locus. 125I-LDL processing values from non-HCs, HC heterozygotes and HC homozygotes differed significantly from one another, but non-HCs and HC heterozygotes showed some overlap. LDL receptor dependent 125I-LDL association (plasma membrane binding plus intracellular accumulation) data for 6 HC heterozygous and 13 non-HC fibroblast strains clustered into two and into three groups, respectively. Median 125I-LDL association levels in these groups appeared to be in agreement with hypothesis that two different geno-types in HC heterozygotes and three in non-HCs determined LDL receptor activity. These findings suggest the possibility that 125I-LDL processing studies may reveal "normal" alleles at the LDL receptor locus. PMID- 6273033 TI - In vitro and in vivo stimulation of neutrophil migration and lymphocyte transformation by thiamine related to inhibition of the peroxidase/H2O2/halide system. AB - The effects of thiamine on neutrophil functions and mitogen-induced lymphocyte transformation were investigated in vitro and in vivo in adult volunteers following the injection of 50 mg thiamine intramuscularly. Thiamine caused stimulation of neutrophil motility in vitro and in vivo and increased lymphocyte transformation in vivo. Enhancement of these functions was related to inhibition of neutrophil post-phagocytic iodination of Candida albicans by the MPO/H2O2/halide system. The horseradish peroxidase/-H2O2/125 I-mediated iodination of bovine serum albumin was also inhibited by thiamine concentrations which caused increased neutrophil motility. It was found that preincubation of neutrophils and lymphocytes with the horseradish peroxidase/H2O2/halide system caused considerable inhibition of the migratory and proliferative responses respectively. Inclusion of thiamine at concentrations which were found to inhibit the peroxidase/-H2O2/halide system protected the neutrophil migratory and lymphocyte proliferative responses from inactivation by this system. It is suggested that thiamine may cause increased neutrophil migration and lymphocyte transformation by protecting these cells from toxic oxidative products generated by the peroxidase/H2O2/halide system. PMID- 6273034 TI - Elevated nephrogenous cyclic adenosine monophosphate to monitor early renal osteodystrophy. AB - Radiographs, serum chemistries, parathyroid hormone (PTH), and nephrogenous cyclic adenosine monophosphate (cAMP) were evaluated in thirty-two children with normal serum creatinine, chronic renal insufficiency, chronic hemodialysis, and transplantation. Nephrogenous cAMP increases linearly with creatinine, and there is a good correlation (r = 0.89) between immunoreactive PTH (iPTH) and nephrogenous cAMP except for patients with severe renal insufficiency or requiring chronic hemodialysis. Elevated nephrogenous cAMP is evidence for metabolic bone disease earlier than usually recognized. Early measurements of iPTH and nephrogenous cAMP could ensure early therapeutic intervention which might alleviate renal osteodystrophy in chronic renal insufficiency and transplant patients. PMID- 6273035 TI - The current concept for the cardiac glycoside receptor. AB - This brief review emphasizes the significance of the Na+,K+-ATPase or the Na+,K+ pump of the intact membrane as the pharmacological receptor for cardiac glycosides. The properties of transport enzyme and the regulation of glycoside binding are described. An outline is given of the problems encountered and of the progress made in attempting to correlate the inotropic action of cardiac glycosides with the binding of these drugs to the heart muscle and with the inhibition of the Na+,K+ pump. Furthermore, the correlation of intracellular Ca2+ activity an Na+ concentration with the inhibition of the Na+,K+ pump is discussed. The existence of a digitalis-like endogenous activity may also indicate an important role of the Na+,K+ pump as a receptor for a physiological regulatory control of cardiac contractility. PMID- 6273036 TI - Vaginal condylomata: a human papillomavirus infection. AB - In our clinic, as a rule, we do not treat vaginal condylomata. They are usually subclinical and asymptomatic. When atypia is present on biopsy, they should be treated in the same manner as vaginal intraepithelial neoplasia. When vaginal discharge and pruritus are present, infection should be searched for and treated. When condylomata are seen with the naked eye, colposcopy has shown that there were many more, too small to be seen, so that local therapy seems a waste of time. If on colposcopic examination only a few condyloma acuminata are located, then therapy is defendable. CO2 laser therapy should be preferred to other modalities until a systemic treatment is available and safe. PMID- 6273038 TI - Current clinical applications of vitamin D metabolite research. AB - Vitamin D2 or D3 are prohormones that are converted in the liver and kidney to three metabolites, all having biological activity. the most prevalent circulating metabolite is 25(OH)D, and when its level falls, osteomalacia and rickets result. The second most prevalent metabolite is 24,25(OH)2D which may be a mineralizing hormone and which may impair PTH secretion, although wide controversy prevails over its function. Calcitriol circulates at levels a thousand-fold less than 25(OH)D and is the most active metabolite in terms of calcium absorption from the gut and mobilization of calcium and phosphate from bone. Clinical defects in this system result from abnormalities of vitamin D metabolism or from end-organ resistance to these hormones. Many disorders of demineralization can now be understood more clearly, and treated with a greater understanding of the underlying pathophysiology. Nonetheless, many new questions about the role of various vitamin D metabolites have now arisen. PMID- 6273037 TI - Early prediction of avascular necrosis of the femoral head following femoral neck fractures. AB - The treatment of acute fracture of the femoral neck remains an unsolved problem. Fifty-three patients are presented using 99mTc phosphate scintigraphy with quantitative computer interpretation to predict the viability of the femoral head following acute fracture. The accuracy of prediction was 92.5%; the scans were incorrect in four patients. Armed with a safe, simple diagnostic procedure and a greater than 90% accurate prognosis, a rational program of treatment can be prescribed for the individual patient. PMID- 6273039 TI - The relevance and future of measurements of regional cerebral blood flow. PMID- 6273040 TI - Megadolicho vascular malformation of the intracranial arteries. AB - A patient is presented suffering a hemiparesis. Megadolicho-vascular malformation of the intracranial part of the internal carotid arteries and some of its branches and of the basilar artery was suggested by CT and confirmed by angiography. The value of CT compared with angiography in relation to intracranial megadolicho vascular malformations is discussed. PMID- 6273041 TI - Wasted leg syndrome: a clinical, electrophysiological and histopathological study. AB - Forty cases of 'wasted leg syndrome' were studied clinically and electrophysiologically. Muscle biopsy was examined in nine cases, majority of patients were adults engaged in heavy manual work. The illness was noticed incidently with a strictly unilateral wasting of the whole lower limb (in 65% of cases), of all muscles below the knee (in 22.5% of cases) or of quadriceps muscles only (in 12.5% of cases). The nerve conduction studies and the electromyographic pattern suggested anterior horn cell disorder. Neurogenic atrophy was seen in 7 out of 9 muscle biopsies. A follow up in 12 patients (2-6 years) revealed no progression of the disease. It is suggested that possibly these cases represent an entity, clinically different from other anterior horn cell disorders. PMID- 6273042 TI - Melanotic rhabdomyomedulloblastoma or teratoid tumour of the cerebellar vermis. AB - Mixed medulloblastomatous tumours are known to comprise several histological characteristics. A three year old boy is reported with a tumour containing elements of medulloblastomatous, rhabdomyosarcomatous and melanotic tissues. PMID- 6273043 TI - Acrylic prosthesis of the fifth cervical vertebra in cervical chordoma. Case report and review of the literature. AB - Description of a two-stage surgical treatment of a case of chordoma of the fifth cervical vertebra mainly affecting the vertebral body. In the first stage the cervical column was immobilized employing a posterior approach and a small portion of the tumour in the right lamina was removed. In the second stage the whole of the body of C5 was removed by an anterior route and replaced with an acrylic prosthesis. The advantages of acrylic resin and the literature on vertebral prostheses are discussed. PMID- 6273044 TI - Orbital bruits and retinal artery pressure in internal carotid artery occlusion. AB - Four patients are reported with orbital bruits. They had minor neurological deficits and all proved to have occlusion of the contralateral internal carotid artery. In three of the patients the retinal artery pressures were normal with no side to side difference, whereas pressure was slightly reduced on the occluded side in one patient. These observations give further evidence that an orbital bruit may indicate occlusion of the contralateral carotid artery with good collateral circulation. PMID- 6273045 TI - Rapid thyroid enlargement: intracystic hemorrhage. PMID- 6273046 TI - Epidemiology of chronic congenital and perinatal infections of man. PMID- 6273047 TI - Eye findings in intrauterine infections. PMID- 6273048 TI - Hepatic manifestations of congenital and perinatal disease. PMID- 6273049 TI - Cardiac disease in congenital infections. PMID- 6273050 TI - Immunologic aspects of congenital infections. PMID- 6273051 TI - Radiological findings in perinatal infections. PMID- 6273052 TI - Diagnosis of viral infections of the newborn infant. PMID- 6273053 TI - Prenatal care in the prevention of infection. PMID- 6273054 TI - Prevention of intrauterine and perinatal infections. PMID- 6273055 TI - Effect of ouabain and furosemide on erythrocyte sodium and phosphate transport. AB - The effects of ouabain and furosemide on the unidirectional efflux of sodium and phosphate ions were studied in freshly drawn human red blood cells (RBCs). In the presence of physiologic concentrations of sodium and potassium the rate of sodium efflux was reduced by 74% due to ouabain sensitivity. Furosemide (1.0 mmol/l) reduced ouabain-insensitive sodium transport rate by a further 50%. Thus, 13% of total sodium efflux was inhibited by furosemide when ouabain was present. In the absence of ouabain, however, furosemide inhibited 31% of total sodium transport, indicating that it also affected ouabain-sensitive sodium efflux. Phosphate transfer of RBCs was almost 1.0 mmol/l RBCs per hour. Erythrocyte concentration of orthophosphate, however, was only 0.59 mmol/l RBCs. Organic phosphate esters must therefore have been cleaved to maintain constant phosphate elimination. The hydrolysis of adenosine triphosphate (ATP) by Na-K-ATPase might be involved because the phosphate transfer of almost 0.12 mmol/l RBCs per hour was ouabain sensitive. Furosemide reduced phosphate efflux by 50% due to reduction in passive permeability of the RBC membrane. Additional inhibition of any phosphate ester hydrolyzing enzymatic activity cannot, however, by excluded. PMID- 6273056 TI - [Experimental lesions of the myocardium: role of alterations of energetic metabolism]. PMID- 6273057 TI - Nonasbestos fibrous minerals. PMID- 6273058 TI - Lung disease secondary to inhalation of nonfibrous minerals. PMID- 6273059 TI - Prolonged neuromuscular blockade after administration of physostigmine and succinylcholine. PMID- 6273060 TI - Death in aftercare. PMID- 6273061 TI - Esophago-pulmonary toxicity from concomitant use of adriamycin and irradiation. PMID- 6273062 TI - Pneumococcal polysaccharide vaccine. Centers for Disease Control, H.E.W., Public Health Service, Atlanta, GA. PMID- 6273063 TI - A steady-state kinetic analysis of the prolyl-4-hydroxylase mechanism. AB - Published kinetic data by Kivirikko, et al. on the prolyl-4-hydroxylase reaction have been re-evaluated using the overall steady-state velocity equation in the forward and reverse directions for an ordered ter ter kinetic mechanism. Qualitatively, the published data for prolyl-4-hydroxylase appear to fit the predicted patterns for this kinetic mechanism. More kinetic data are needed to confirm these results and to quantitate the kinetic parameters but, tentatively, the order of substrate addition would appear to be alpha-ketoglutarate, oxygen, and peptide; and the order of product release would be hydroxylated peptide (or collagen), carbon dioxide, and succinate. PMID- 6273064 TI - Win 32,729, a new, potent interceptive agent in rats and rhesus monkeys. AB - Win 32,729 [(2 alpha, 4 alpha, 5 alpha, 17 beta)-4,5-epoxy-17-hydroxy-4,17 dimethyl-3-oxoandrostane-2-carbonitrile] is an orally active interceptive agent in rats and rhesus monkeys (M mulatta). A single oral dose of 48 mg/kg terminated gestation when given on Day 10 of pregnancy. When given orally for up to 5 days to pregnant monkeys, it terminated pregnancy in 26 of 34 animals at a dose of 50 mg/monkey (ca 7 mg/kg), in 18 of 24 at a dose of 100 mg/monkey (ca 14 mg/kg) and in all 6 at 250 mg/monkey (ca 35 mg/kg). It did not inhibit ACTH-stimulated glucocorticoid production at 50 mg/monkey but did at a dose of 250 mg/monkey. This preferential gonadal inhibition was not evident in rodents. While in most cases five oral medications of 50-100 mg were required to terminate gestation in 50-day pregnant monkeys, a single subcutaneous medication with 250 mg was also effective, terminating pregnancy in 7 of 7 monkeys. PMID- 6273065 TI - Cryoenzymology. AB - A detailed understanding of the mechanism of enzyme action requires a correspondingly detailed knowledge of the structures of the intermediates and transition states on the catalytic pathway, as well as the kinetics and thermodynamics of their interconversion. Cryoenzymology, the use of subzero temperatures and fluid cryosolvents, has the potential to supply this type of information. In this article recent investigations illustrating the advantages of cryoenzymology are reviewed. The major advantage lies in the ability to accumulate and stabilize productive enzyme-substrate intermediates for sufficient lengths of time to allow the collection of high resolution structural data, e.g. by X-ray diffraction. In addition, intermediates which are not detectable under normal conditions, due to too short lifetimes and/or low concentrations, may be detected at low temperatures, in some cases through changes in the rate-limiting step. The method seems of rather general applicability, judging by the number of different types of enzymes, including oligomeric and membrane-associated ones, which have successfully been studied using cryoenzymology. In some cases, at least, good agreement between observations at subzero temperatures and normal conditions has been found, demonstrating the relevance of the technique. Potential limitations of the techniques, as well as questions regarding the effects on the protein structure, are also discussed. PMID- 6273066 TI - Nonspecific local immune inhibition of herpes simplex virus keratitis. AB - Rabbit cornea, which had recovered from initial sensitization to bovine serum albumin were found to possess immune memory capable of responding to a totally different antigenic stimulus, i.e., viable herpes simplex virus. Two months after BSA sensitization these corneas were found to be extremely resistant to primary infection by the virus. The long term residence of mononuclear leukocytes in the cornea, following BSA sensitization, is apparently responsible for this non specific immune phenomenon. PMID- 6273067 TI - Restriction endonuclease cleavage of DNA obtained from herpes simplex isolates of two patients with bilateral herpetic disease. AB - Restriction enzyme analysis was utilized to investigate HSV isolates from bilateral lesions of skin and/or cornea. In the first case (DM) type 1 HSV was obtained from a lesion on the right eye lid in 1977 and from a recurrence in 1979 consisting of dendritic lesions on the left cornea as well as skin lesions on the right naris. HSV virus was isolated in the second case (BL) from a right corneal lesion in 1979. Ten months later HSV was again isolated from a recurrence on the right cornea and from multiple skin lesions on the left cheek. DNA obtained from all HSV isolates were cleaved with Hpa I, Bgl II, Kpn I, Hind III, and Eco RI. A comparison of DNA cleavage patterns following agarose electrophoresis revealed that the virus isolates from DM were different from those of BL. However, all of the isolates obtained from each patient were identical. Therefore, these type 1 isolates obtained from anatomical sites supplied by contralateral trigeminal nerves were due to reactivation and/or reinfection with endogenous virus rather than exogenous infection. PMID- 6273068 TI - Some problems in modern anti-ulcer drug evaluation: a comparison of four gastric acid suppressors. AB - Some of the problems involved in the treatment and evaluation of drug therapy in peptic ulceration are discussed. The results of four multi-centre, double-blind, placebo-controlled clinical trials in 520 patients are reviewed. It has been shown that the potent suppressors of gastric acid secretion, histamine H2 blockers ranitidine and cimetidine, and 16,16-dimethyl prostaglandin E2, significantly accelerated the healing of peptic ulcers but that trithiozine had no beneficial effect. PMID- 6273069 TI - The transforming protein of avian sarcoma viruses and its homologue in normal cells. PMID- 6273070 TI - Gene-specific probes for avian retroviruses. PMID- 6273071 TI - Replication of herpesvirus DNA. PMID- 6273072 TI - Differentiation and viral involvement in differentiation of transformed mouse and rat erythroid cells. PMID- 6273073 TI - Catalase levels in patients with aniridia and/or Wilms' tumor: utility and limitations. AB - The gene for red blood cell (RBC) catalase has recently been mapped to 11p13, and a gene dosage effect has been demonstrated for individuals with triplication or deletion of that region. Deletion of the 11p13 band has also been associated with aniridia, with and without Wilm's tumor. We studied the RBC catalase levels in individuals without detectable chromosomal abnormalities but with aniridia, Wilm's tumor, and the combination of aniridia and Wilms' tumor, to determine whether catalase levels might provide evidence for a submicroscopic chromosomal deletion in the 11p13 region. All karyotypically normal patients were found to have normal catalase levels. PMID- 6273074 TI - The preferential deposition of inhaled isoproterenol and propranolol in asthmatic patients. AB - This study describes the use of central and diffuse airway deposition patterns of isoproterenol and radiotracer aerosol alone, and in the presence of centrally deposited propranolol and radiotracer aerosol, to investigate the distribution of beta 2 adrenoreceptors in six asthmatic patients. The central deposition technique was only partially successful. No definite beta 2 receptor distribution pattern could be interpreted from the airway function responses. It was noted that 3-10 micrograms of isoproterenol in the airways was able to produce 50 percent of the maximum possible flow rate response. Centrally deposited propranolol was an effective antagonist of isoproterenol. PMID- 6273075 TI - Orbital chemodectoma. Clinical and pathologic analysis of 2 cases. PMID- 6273076 TI - Malignant bone fibrous histiocytoma. Report of 3 cases. PMID- 6273077 TI - [Klippel-Trenaunay disease]. PMID- 6273078 TI - [The suprascapular nerve syndrome]. PMID- 6273080 TI - [Granulosa and theca cell tumor---clinicopathologic analysis of 57 cases (author's transl)]. PMID- 6273079 TI - [Contribution of radiology to the postoperative after care of patients with colorectal tumors]. AB - The prognosis of local recurrences after surgery for colorectal cancer is better than in recurrences of many other tumor sites. Since in most cases secondary tumor growth takes place as local recurrence and distant metastases are infrequent, there is a good chance of detecting the recurrence early in a curable state. This offers good conditions for a radical second operation. Following excision of tumors, double-contrast enema permits a very detailed assessment of the anastomosis, allowing detection of small local recurrences and recognition of postoperative changes or sequelae of delayed healing at the site of the anastomosis. Following amputation of the rectum, computed tomography permits better control of the presacral space than other conventional radiodiagnostic procedures. In both situations, however, integration of both methods is necessary in a consequent time schedule of clinical follow-up with suitable intervals. PMID- 6273081 TI - [Preliminary observation of the variation of plasma cyclic adenosine monophosphate (C-AMP) in cor pulmonale (author's transl)]. PMID- 6273082 TI - [Lecture: the interrelation between endocrinology and clinical internal medicine (author's transl)]. PMID- 6273083 TI - [Experimental studies on the isolation of enteric viruses from hospital sewage by polymer two-phase method (author's transl)]. PMID- 6273085 TI - [The cytotoxic effect of lymphocytes from patients with nasopharyngeal cancer (NPC) against a NPC epitheloid cell line (author's transl)]. PMID- 6273084 TI - Intestinal absorption of triglyceride and vitamin D3 in aged and young rats. AB - [3H]Trioleyl glycerol (TO) and [14C]vitamin D3 were perfused intraduodenally for 5 hr in aged (19-21 months) and young adult (4-5 months) Sprague-Dawley rats. The rate of intestinal uptake from the gastrointestinal lumen and transport into the body of these lipids were decreased in the aged animals. Since the distribution of TO lipolytic products in the lumen was unchanged, reduced intestinal uptake rate probably occurred at the mucosal membrane. Furthermore, in the aged rats, the rate of transintestinal transport of both trioleyl glycerol and vitamin D3 was impaired. No evidence for impaired mucosal TO reesterification or for accumulation of vitamin D3 metabolites was found, suggesting that intestinal lipid accumulation resulted from a defect in lipoprotein assembly or in discharge from the mucosal cell. Impaired absorption of lipids may contribute to malnutrition and osteopenia of advancing age. PMID- 6273086 TI - [Lymph scan with radionuclides in gynecologic tumors (author's transl)]. PMID- 6273087 TI - [Surgical treatment of primary hepatoma--a report on 117 cases (author's transl)]. PMID- 6273088 TI - [Insulinoma (author's transl)]. PMID- 6273090 TI - [Model of a "complex" receptive field in a visual cortex neuron]. PMID- 6273089 TI - [Actinomycin D derivatives that recognize 2 DNA GC pairs: bis-actinomycin D]. PMID- 6273091 TI - [Ion channels in bilayer lipid membranes dependent on oxygen]. PMID- 6273092 TI - [Specific binding of a low-molecular factor with the Fc fragment of human immunoglobulin G]. PMID- 6273093 TI - [A sensitive method to determine cyclic adenosine monophosphate in platelets (author's transl)]. PMID- 6273094 TI - [Preliminary clinical use of radionuclide myocardial imaging (author's transl)]. PMID- 6273095 TI - [Effects of norepinephrine on the activity of Na+-K+-ATPase and level of cAMP in rabbit myocardium and the antagonistic effects of insulin (author's transl)]. PMID- 6273096 TI - [Surgery of bronchial carcinoma]. PMID- 6273097 TI - [Computed tomographic brain scanning in the diagnosis of metastatic neoplasms (author's transl)]. AB - Clinical investigations and computed brain scanning were done in 305 patients with primary extracerebral malignant tumours. One third of the patients had cerebral metastases. In most patients with brain metastases extracerebral secondary tumours were known already. Silent brain metastases were present in only 0.6% of all investigated tumour patients. All other patients had either objective neurologic-psychiatric defects or a least symptoms (headache, vomiting). Use of cranial computed tomography in all tumour patients as a pure screening method is thus not justified. The indication for the investigation is dependent on the clinical symptomatology. However, not only objective neurologic psychiatric defects must be taken into account, but also occurrence of new symptoms. PMID- 6273098 TI - [Comparison of the morphological and serological findings in maedi-virus infection of sheep (author's transl)]. PMID- 6273099 TI - [Experimental intranasal infection of cattle with Aujeszky's disease virus (author's transl)]. PMID- 6273100 TI - [Comparative studies of porcine sera with an immunoenzyme test (ELISA) and neutralization test for serodiagnosis of Aujeszky's disease in swine]. PMID- 6273101 TI - Cell culture systems for studying multifactor interactions in carcinogenesis. AB - Short term assays for potential carcinogenic factors must take into account the fact that carcinogenesis is usually a multistep process and that most human cancers probably result from a complex interaction between multiple factors, both environmental and endogenous. Thus, although extremely valuable, simple in vitro mutagenesis assays have major limitations. Cell culture studies with TPA and other phorbol esters provide clues to tumor promotion and the multistep nature of carcinogenesis. These effects can be divided into three categories: 1) mimicry of transformation in normal cells, and enhancement of transformation by chemical carcinogens or oncogenic viruses; 2) modulation (inhibition or induction) of differentiation; 3) membrane and receptor effects. Current evidence suggests that TPA acts by binding to specific high affinity cell surface membrane receptors and that this leads to alterations in the composition of membrane phospholipids. Presumably, these changes in the lipid matrix of cell membranes produce signals or mediators which lead to the subsequent cytoplasmic and nuclear effects of TPA. Thus, whereas the critical target in the action of initiating carcinogens appears to be cellular DNA, the critical target of the phorbol ester tumor promoters appears to be cell membranes. As a unifying concept of two-stage carcinogenesis, we postulate that during the initiation phase in carcinogenesis the covalent binding of carcinogens in DNA induces abberations in the commitment of the target cells. We believe that this involves highly ordered genetic events, for example, gene transpositions, rather than random point mutations. Certain oncogenic viruses achieve a similar effect thru integration of exogenous DNA sequences into inappropriate sites in the host genome. Tumor promoters, via their effects on growth, gene expression and differentiation, enhance the selective outgrowth of these initiated cells and induce them to express their newly acquired by previously dormant committed state. Rapid in vitro assays for the detection of the phorbol ester tumor promoters and for synergistic interactions between oncogenic viruses and chemical now exist. However, simple assays for other classes of tumor promoters and cofactors remain to be developed. PMID- 6273102 TI - Effect of glutathione and uridine 5'-diphosphoglucuronic acid on benzo(a)pyrene mutagenesis in the Salmonella/microsome assay. PMID- 6273103 TI - Metabolic inactivation of mutagenic benzo(a)pyrene metabolites: significance to carcinogenicity and implications for in vitro tests. PMID- 6273104 TI - Synergistic effects of cigarette smoke extracts, benz (a) pyrene and nickel sulphate on morphological transformation of hamster embryo cells. PMID- 6273105 TI - Recent morphological studies of toxic neuropathy. PMID- 6273107 TI - Brain slices, assay systems for the neurotoxicity of environmental pollutants and drugs on mammalian central nervous system. PMID- 6273106 TI - Neurotoxic effects of long-term exposure to organic hydrocarbon solvents. Epidemiologic aspects. PMID- 6273108 TI - Biochemical changes in the rat cerebellum following cypermethrin administration. PMID- 6273109 TI - A sensitive experimental model for the assessment of hepatotoxicity by halogenocompounds: prolactin binding to the specific receptors. PMID- 6273110 TI - Sensitivity of rat brain synaptosomal ATPases to several structurally related organochlorine compounds. PMID- 6273111 TI - Diagnosis and management of less common causes. PMID- 6273112 TI - The management of Wilms' tumour in Kenya. PMID- 6273113 TI - Neurotoxicity in relation to tropical peripheral neuropathy: a re-appraisal. PMID- 6273114 TI - [Tumor scintigraphy of patients with malignant tumors in the head and neck area]. PMID- 6273115 TI - [Angiotensin receptors]. PMID- 6273116 TI - [Effect of tyrosine-74-modified cytochrome C on electron transfer in Keilin Hartree submitochondrial particles]. AB - The tyrosine-74 modified cytochrome c is obtained by the interaction of cytochrome c with N-(2, 2, 5, 5-tetramethyl-3-carboxy-piroline-oxyl) Imidazol. The absorbtion spectrum of the modified derivative at the visible region does not differ substantially from the native protein. The band of absorbtion at 695 nm, which shows that the performed modification is soft and the conformation of the modified cytochrome c is close to that of the native protein. Redox potential of the modified cytochrome c, determined by spectrophotometric titration, does not differ from that of the native protein. Submitochondrial particles, deficient of cytochrome c, are reconstructed with modified protein. In both substrates of biologic oxidation succinate and ascorbate the electron transport is restored by the modified protein in considerably higher concentrations than those of the native cytochrome c. Inspite of small conformation differences and unchanged redox potential of the modified cytochrome c substantial differences are observed in its electron-carrying properties as well as its interaction with the membrane cytochrome c reductase and oxydase. PMID- 6273117 TI - Mineralocorticoid receptors in the rat lung. AB - In lung cytosol from adrenalectomized rats, tritiated aldosterone binds to two classes of limited capacity sites with affinities (Kd at 37 C) of 0.7 and 80 nM. The predominant binding species sediments at approximately 9S on sucrose density gradients in low ionic strength, molybdate-containing buffer. Binding of tritiated aldosterone (5 nM) is displaced by a 4-fold excess of nonradioactive hormone in the following hierarchy: 9 alpha-fluorocortisol greater than aldosterone greater than deoxycorticosterone much greater than corticosterone greater than dexamethasone. Thus, pulmonary tissue contains steroid-binding sites with affinity and specificity characteristics equivalent to those previously shown for mineralocorticoid receptors in classic target tissues. The pulmonary cell type(s) in which these putative mineralocorticoid receptors are located and possible physiological roles for mineralocorticoids in adult and fetal lung remain to be explored. PMID- 6273118 TI - Differential processing of the two subunits of human choriogonadotropin (hCG) by granulosa cells. I. Preparation and characterization of selectively labeled hCG. AB - The alpha- and beta-subunits of hCG were radioiodinated and recombined with unlabeled complementary subunits. The resultant recombined hormones, selectively labeled in either the alpha- or beta-subunit, were separated from unrecombined subunit by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, extracted with Triton X-100, and characterized by binding analysis. The estimates of maximum binding (active fraction) of the two resultant selectively labeled, recombined hCG preparations, determined with excess receptor were 0.41 and 0.59. These values are similar to those obtained when hCG is labeled as an intact molecule. The specific activities of the recombined preparations were estimated by four different methods, and the resulting values were used in combination with the active fraction estimates to determine the concentrations of active free and bound hormone. Binding analyses were run using varying concentrations of both labeled and unlabeled hormone. Estimates of the equilibrium dissociation binding constant (Kd) and receptor capacity were calculated in three different ways. The mean estimates of capacity (52.6 and 52.7 fmol/mg tissue) and Kd (66.6 and 65.7 pM) for the two preparations were indistinguishable. Additionally, these values were similar to values reported previously for hCG radioiodinated as an intact molecule. The availability of well characterized, selectively labeled hCG preparations provides new tools for studying the mechanism of action and the target cell processing of the subunits of this hormone. PMID- 6273120 TI - Adrenocorticotropin and adenosine 3',5'-monophosphate stimulate de novo synthesis of adrenal phosphatidic acid by a cycloheximide-sensitive, CA++-dependent mechanism. AB - We tested further our postulate that enhanced de novo synthesis of phosphatidic acid is responsible for ACTH- and cAMP-induced increases in adrenal phospholipids in the phosphatidate polyphosphoinositide pathway. During incubation of adrenal sections or cells in vitro, ACTH and cAMP increased the concentrations of and incorporation of [3H]glycerol and [14C]palmitate into phosphatidylcholine and phosphatidylethanolamine, two major phospholipids which are derived from phosphatidic acid, but are extrinsic to the inositide pathway. Thus, it is unlikely that ACTH and cAMP increase inositide phospholipids at the expense of other phospholipids. Similar to previously reported effects on phosphatidic acid and inositide phospholipids, cycloheximide blocked the effects of ACTH and cAMP on phosphatidylcholine and phosphatidylethanolamine. In addition, Ca++ was required for these effects, as well as for cAMP-induced increases in phosphatidic acid, inositide phospholipids, and steroidogenesis. Our findings strongly suggest that ACTH, via cAMP, stimulates de novo phosphatidate synthesis by a cycloheximide-sensitive, Ca++-dependent process, and this stimulation causes a rapid generalized increase in adrenal phospholipids. Moreover, the increased incorporation of labeled glycerol and palmitate into phospholipids suggests that ACTH and cAMP may stimulate the glycerol-3'-PO4 acyltransferase reaction. This stimulatory effect may play a central role in the steroidogenic and trophic actions of ACTH and cAMP. PMID- 6273119 TI - Differential processing of the two subunits of human choriogonadotropin by granulosa cells. II. In vivo studies. AB - Using a well characterized preparation of hCG, consisting of a mixture of hCG labeled in the alpha-subunit with 125I and hCG labeled in the beta-subunit with 131I (see preceding paper), the hormone-specific preferential retention by granulosa cells in vivo of the radiolabel originally associated with the beta subunit of hCG has been confirmed and extended. Additional studies have shown that this retention is peculiar to the granulosa cells. Luteal and interstitial/thecal elements of the ovary failed to show preferential accumulation of the label originally associated with the beta-subunit. Measurement of both radioactivities in crude subfractions of the ovarian tissues revealed that granulosa cells retain the excess of beta-subunit label in a plasma membrane/vesicular component. No such preferential retention of label was seen in any of the subfractions obtained from luteal or interstitial/thecal tissues. The radiolabeled components associated with the granulosa cells were shown to be mainly macromolecular by their precipitability with 13% trichloroacetic acid. Luteal tissue degraded the components associated with each label more rapidly than granulosa cells. In contrast, interstitial/thecal tissue degraded very little of the bound labeled components. The differential processing of individual hCG subunits by granulosa cells was shown not to result from different kinetics of binding of serum-borne hormone by two methods. Thus, changes over time in the ability of circulating hormone to bind to LH receptor in vitro were shown not to be a function of the hCG subunit having the label. Moreover, blockade of further radiolabel uptake by injection of a large excess of unlabeled hCG 30 min after radiolabel administration did not alter the rise in the ratio of beta-subunit label to alpha-subunit label normally observed in granulosa cells. The ability of kidney tissue to accumulate and metabolize hCG also varied with the physiological state. Within the limitations of following the radioiodides added to proteins rather than the peptides themselves, these studies demonstrate that differences exist in the metabolism of hCG by the various target cells of the ovary and that changes in processing occur during luteinization. PMID- 6273121 TI - Catechol estrogen formation by brain tissue: a comparison of the release of tritium from [2-3H]estradiol with [6,7-3H]2-hydroxyestradiol formation from [6,7 3H]estradiol by rabbit hypothalami in vitro. AB - In the course of establishing an assay for estrogen-2-hydroxylase activity, a detailed comparison was made between the formation of tritiated water (3H2O) and [6,7-3H]2-hydroxyestradiol (2-OHE2) by rabbit hypothalami in vitro from 2-3H- and 6,7-3H-labeled estradiol, respectively. The amounts of both 3H2O and [6,7-3H]2 OHE2 formed were stimulated several-fold by the nonionic detergent Tween-80. Maximum activity for both functions was associated with the soluble fractions (S2, 17,500 X g supernatant, for tritium release; S3, 100,000 X g supernatant, for 2-OHE2 formation). In contrast, maximal 3H2O formation by rat liver was associated with the microsomal (P3, 100,000 X g pellet) fraction and was virtually abolished by Tween-80. The amount of 3H2O formed exceeded, up to severalfold, the amount of 2-OHE2 produced under all conditions examined and in all subcellular fractions. The bulk of the excess 3H2O formation, unrelated to the production of 2-OHE2, could be eliminated by adding ascorbic acid (10 mM) to the incubation medium. However, a second, smaller component of spurious 3H2O release could not be suppressed. This component was responsible for a persistent lack of stoichiometry between the formation of 3H2O and 2-OHE2, with the former exceeding the latter by up to 2-fold. This discrepancy was unaffected by ascorbic acid (up to 20 mM), unlabeled 2-OHE2 (up to 10 microM), and reducing the temperature of incubation from 37 to 30 C, measures that prolonged the t1/2 of 2 OHE2 during incubation with hypothalamic tissue from under 3 min to over 100 min. These findings 1) raise doubts about the validity of using 3H2O formation from [2 3H]estradiol as a quantitative index of estrogen-2-hydroxylase activity, and 2) establish conditions under which further metabolism of 2-OHE2 is inhibited, thereby making it practical to quantify enzyme activity on the basis of the amount of catechol estrogen formed. Evidence is also presented that the release of 3H2O from [2-3H]estradiol by hypothalamic tissue, unrelated to 2-OHE2 formation, may be enzymatically mediated. PMID- 6273122 TI - Catechol estrogen formation by brain tissue: characterization of a direct product isolation assay for estrogen-2- and 4-hydroxylase activity and its application to studies of 2- and 4-hydroxyestradiol formation by rabbit hypothalamus. AB - A direct product isolation assay for quantifying the formation of 2- and 4 hydroxyestradiol (2-OHE2 and 4-OHE2) from [6,7-3H]estradiol by rabbit hypothalami in vitro was developed, and the assay was used to characterize some properties of estrogen-2- and 4-hydroxylase activity in this tissue. The reaction was carried out under conditions that minimized further metabolism of enzymatically formed catechol estrogens. A simple two-step separation procedure, involving the use of a neutral alumina column, followed by thin layer chromatography, was developed to isolate the enzymatically formed catechol estrogens in a radiochemically homogeneous form. The detergent, Tween-80, was found to activate the enzyme and was used routinely at a concentration of 0.1% in the assay. The formation of 2 OHE2 was linear up to 10 min and with increasing protein concentrations up to 150 micrograms/incubation. Similar values were obtained for 4-OHE2. Maximum velocities (Vmax) for the formation of 2- and 4-OHE2 were 190 and 270 pmol/mg protein . 10 min, respectively. The apparent Km values with respect to estradiol for 2-OHE2 and 4-OHE2 were 125 and 150 microM, respectively. The highest specific activity for the enzyme was present in the 100,000 X g supernatant (S3), while the activity in the microsomal fraction (P3) was less than that in the original homogenate. Enzyme activity depended on the presence of NADPH and oxygen and was inhibited by CO as well as by high concentrations of SKF-525A. Estrogen-2- and 4 hydroxylase activity in rabbit hypothalamus differed from that in rat liver in two respects. In the liver, enzyme activity was localized in the microsomal fraction and was virtually abolished by Tween-80. In contrast, enzyme activity in rabbit hypothalamus was maximal in the soluble fraction (100,000 X g supernatant)and was stimulated by the detergent. PMID- 6273123 TI - Contribution of subunits to the function of luteinizing hormone/human chorionic gonadotropin recombinants. AB - The alpha- and beta-subunits of hCG and of ovine and porcine LH were used to prepare all nine homologous and heterologous alpha beta-recombinants. EAch purified recombinant was assayed in vitro, using dispersed Leydig tumor cells derived from the M5480P tumor, for its ability to stimulate steroidogenesis and to inhibit [125I]iodo-hCG binding. It was found that the potency of a given recombinant in both assays was most similar to that of the hormone from which the beta-subunit was derived. For example, hCG and hCG beta-containing recombinants were invariably more potent than LH and LH beta-containing recombinants. However, within groups of recombinants containing a common beta-subunit, the alpha-subunit exhibited modulatory effects on the biological potencies. The different observed potencies did not result from alpha beta dissociation since the recombinants were stable in dilute solution at 37 C for periods greatly exceeding that of the assay conditions. LH and the LH beta-containing recombinants were found to dissociate more readily from the Leydig tumor cell gonadotropin receptor than hCG and hCG beta-containing recombinants. (These experiments were performed under conditions where internalization was minimal.) However, analogous to the potency measurements, the alpha-subunit contributed to the rate of dissociation. For example, in recombinants with a common beta-subunit, hCG alpha conferred the greatest stability to the hormone-receptor interaction. These results emphasize a positive relationship between receptor occupancy and biological potency. Whereas the beta-subunit of these gonadotropins seems to exhibit the predominant influence in determining potency, it is clear that both subunits contribute to biological activity. This could involve direct effects as well as induced conformational changes in the complementary subunit. PMID- 6273124 TI - Mineralocorticoid receptor-like aldosterone-binding protein in cell culture. PMID- 6273125 TI - Disproportionate accumulation of immunoreactive corticotropin, melanotropin, and lipotropin in the brain of the maturing rat. AB - The accumulation of immunoreactive ACTH (ACTHi), alpha MSH (alpha MSHi), and gamma-lipotropin (gamma LPHi) as a function of age (10-120 days) was determined in three regions of the brain of male rats: the medial basal hypothalamus (MBH), the preoptic anterior hypothalamus (POA), and the thalamus. In each region of the brain, the concentrations of ACTHi, alpha MSHi, and gamma LPHi increased with age. In the MBH, the increase occurred in such a manner that the molar ratio of alpha MSHi to ACTHi remained constant regardless of the age of the animals. In contrast, in the POA and thalamus, the increase occurred disproportionately in favor of alpha MSHi, and thus the molar ratio of alpha MSHi to ACTHi was 3 times higher in the adult (120 days old) than in the young (10 or 21 days old) animals. Nevertheless, the ratio of (ACTHi plus alpha MSHi) to gamma LPHi was constant at a level of about 2 regardless of the age of the animal or the region of the brain. Extracts of the MBH or POA were fractionated on columns of Sephadex G-75 superfine. The gel filtration profiles of ACTHi were indicative of the presence of five molecular weight forms of ACTH: greater than 40K, 30-40K, 20-30K, 5.7K, and 4.5K. We tentatively identified greater than 40K ACTH as a large form of proopiocortin, 30-40K ACTH as proopiocortin, 20-30K ACTH as ACTH biosynthetic intermediate, 5.7K as glycosylated ACTH(1-39), and 4.5K ACTH as ACTH-(1-39). Regardless of the age of the animals, the fractional amount of 30-40K ACTH the age of the animals, the fractional amount of 30-40K ACTH was high in the MBH compared to that in the POA. Moreover, the small fractional amount of 30-40K ACTH in the POA was associated with a large fractional amount of small molecular weight forms of ACTHi. However, the predominant form of ACTHi in the POA changed with age: 20-30K ACTH was the major form in the young, whereas 4.5K ACTH was the major form in the adult. These results support the proposal that the production of proopiocortin increases with age, and there is enhanced processing of proopiocortin to ACTH-(1-39) and alpha MSH in the brain of the maturing rat. PMID- 6273126 TI - Luteinizing hormone release and gonadotropin-releasing hormone (GnRH) receptor internalization: independent actions of GnRH. AB - Three different approaches are described which provide independent and new evidence that gonadotropin-releasing hormone (GnRH) internalization and GnRH stimulated LH release are distinct actions of the releasing hormone. 1) Removal of GnRH from medium bathing the pituitary cell cultures resulted in the prompt return of LH release to basal levels. This finding indicated that a continuous supply of externally applied GnRH is required for the stimulation of LH release. 2) Covalent immobilization of D-Lys6-des-Gly10-Pro9-ethylamide GnRH (a GnRH agonist) on agarose beads resulted in a derivative which stimulated LH release with full efficacy. At concentrations of immobilized releasing hormone analog sufficient to evoke gonadotropin release, the quantity of LH release was restricted by the number of beads added. This finding was interpreted as evidence that the attachment of immobilized agonist was stable during the bioassay and indicated that LH release could be stimulated with full efficacy without the requirement for GnRH internalization. 3) Comparative studies using image intensified microscopy and the cell culture bioassay showed that 100 microM vinblastin markedly inhibited large scale patching and capping of the GnRH receptor (viewed by image-intensified microscopy), but did not alter the EC50 or efficacy of LH release stimulated by GnRH or the agonist described above. These observations indicated that internalization as well as large scale patching and capping of the GnRH receptor are not required for LH release. PMID- 6273127 TI - Lipolytic effects of insulin in adipocytes isolated from hypophysectomized rats. AB - Adipocytes isolated from the epididymal fat of hypophysectomized rats by digestion with collagenase failed to respond to insulin with an increase in glucose utilization. These cells also exhibited an anomalous response to insulin when added in the presence of epinephrine. While insulin antagonized the lipolytic actions of epinephrine in normal adipocytes or in segments of epididymal fat from normal or hypophysectomized rats, it potentiated lipolysis in adipocytes isolated from hypophysectomized rats. This anomalous effect was also evident when isoproterenol, ACTH, or glucagon was used as the lipolytic agent, but the expected antilipolytic response was obtained when theophylline served as the lipolytic agonist. The lipolytic effects of insulin were not seen until 4-7 days after hypophysectomy. Treatment of hypophysectomized rats with a combination of GH (100 micrograms/rat . day), cortisone acetate (1 mg/rat . day), and T3 (1 micrograms/rat . day) for 5 days restored the antilipolytic response to insulin in cells of hypophysectomized rats, but no one hormone alone was effective. The data indicate that adipocytes of hypophysectomized rats retain their ability to recognize and response to insulin, but the ability of insulin to stimulate glucose oxidation or antagonize epinephrine-induced lipolysis is abolished by the cell isolation procedure. The findings underscore the need to consider the impact of hormonal status on the ability of cells to retain normal responsiveness during the rigors of the cell isolation procedure and suggest that failure to do so might lead to erroneous interpretations of the physiological actions of hormones. PMID- 6273128 TI - Potentiation of epinephrine-induced lipolysis by catechol estrogens and their methoxy derivatives. PMID- 6273129 TI - Inhibition of beta-adrenergic responsiveness in muscle cell cultures by dexamethasone. AB - Cultures of two myogenic cell lines, L6E9 and L8, were grown in the absence or presence of dexamethasone. Dexamethasone (1 microM) completely inhibited the formation of myotubes. Partial inhibition (20-40%) was obtained at a concentration as low as 1 nM. Dexamethasone also inhibited beta-adrenergic responsiveness, as noted by decreases in isoproterenol-stimulated adenylate cyclase activity and cAMP accumulation. These effects were both dose and time dependent. In the presence of dexamethasone, the number of beta-adrenergic receptors, as assessed by [125I]iodohydroxybenzylpindolol binding, decreased coordinately with the decrease in cAMP. A high affinity, limited capacity cytosolic binding site for [3H]triamcinolone acetonide observed under control conditions (Kd = 0.7 nM; maximum binding, 2.7 pmol/mg protein) increased in number as a function of developmental state. These data indicate that glucocorticoids inhibit myogenesis and beta-adrenergic responsiveness in vitro. PMID- 6273130 TI - Spontaneous and adrenocorticotropin (ACTH)-induced maturation of the responsiveness of ovine fetal adrenal cells to in vitro stimulation by ACTH and cholera toxin. PMID- 6273131 TI - Binding of [3H]dihydroergocryptine to an alpha-adrenergic site in the stalk median eminence of the steer. AB - Dihydroergocryptine (DHE), a potent dopamine agonist and alpha-adrenergic antagonist, has been used as a radioligand to characterize both dopamine and alpha-adrenergic receptors. In the present study, the binding of [3H]DHE to particulate fractions of the steer stalk median eminence was characterized using a filtration assay. Specific binding was defined by the presence of 10 microM phentolamine or by an iterative nonlinear hyperbolic curve-fitting program. Scatchard analysis of equilibrium isotherms of specific binding defined a single high affinity (Kd = 1.78 +/- 0.22 nM), saturable (maximum binding, 481 +/- 39 fmol/mg protein), stereoselective binding site. The Kd, calculated from the ratio of the rate constants k2 and k1, was 2.8 +/- 0.14 nM. The rank order of potency of agonists to compete for [3H]DHE binding (l-epinephrine greater than l norepinephrine greater than dopamine greater than l-isoproterenol) was consistent with interactions at an alpha-adrenergic site. The rank order of potency of alpha antagonists (phentolamine greater than yohimbine greater than prazosin) suggested that this was an alpha 2-adrenergic receptor. The affinity of dopamine agonists for the [3H]DHE-binding site was 10-fold lower relative to their potency at known dopamine receptors, while the affinity of dopaminergic antagonists was 100-fold lower. Furthermore, Scatchard analysis of specific [3H]DHE binding in the presence of a concentration of spiperone which should saturate dopamine receptors, only decreased the number of binding sites by 9%. These data demonstrate the presence of large numbers of alpha-adrenergic receptors in the stalk median eminence of the steer. Only a small number of dopaminergic binding sites for [3H]DHE appeared to be present. PMID- 6273132 TI - Role of the sympathetic nervous system in the control of the goitrogenic response in the rat. PMID- 6273133 TI - Induction of insulin-like responses to growth hormone by stress. PMID- 6273134 TI - Hormonal effects on the immunocytochemical location of 3',5'-cyclic adenosine monophosphate-dependent protein kinase in rat tissues. AB - Homogeneous preparations of type I and type II regulatory subunits (RI and RII, respectively) of cAMP-dependent protein kinase (cAMP kinase) were utilized as antigens to obtain isozyme specific antisera. Injections of pure catalytic subunit (C) from the type I isozyme resulted in antisera that reacted with C subunit obtained from either isozyme type. Cross-reactivity of the antisera raised against isolated subunits of the kinase was assessed by immunodiffusion analysis and by measuring the cAMP binding and phosphotransferase activities of the subunits after immunoprecipitation. These antisera were used to localize subunits of type I and type II cAMP kinases in rat skeletal muscle, liver, and adrenal by using indirect immunofluorescence and immunoperoxidase techniques. Specificity of the immunofluorescence was shown by absorption of the antisera with pure homologous antigens. In skeletal muscle, both R and C subunits of the type I and type II cAMP kinases were localized in the area of the sarcoplasmic reticulum and in periodic crossbands. Specific fluorescence for these components was observed in both isotropic and anisotropic band regions of the sarcomere. Densitometric determinations of immunoperoxidase staining revealed a larger amount of RI, RII, and C subunits in the isotropic band than in the anisotropic band regions. In liver, C, RI, and RII subunits were distributed both in cytoplasmic and nuclear areas and along plasma membranes of hepatocytes; however, there were qualitative differences observed among these various subcellular sites. With each antiserum, fluorescence was blocked by prior absorption with homologous antigen. After treatment of rats with glucagon, dramatic changes in the relative distribution patterns of C and RII were noted in the nucleus. In the adrenal gland, RI, RII, and C subunits were localized in both cytoplasmic and nuclear areas, and an apparent redistribution of these subunits occurred after treatment of (dexamethasone-suppressed) rats with ACTH. The application of this immunocytochemical approach provides a tool for examining and monitoring the subcellular distribution of these components of cAMP kinase in biological systems. PMID- 6273135 TI - The role of tubulin in the steroidogenic response of murine adrenal and rat Leydig cells. AB - The Y-1 murine adrenal and CCL43 rat Leydig tumor cell lines were used as model systems for studying the role of tubulin in steroidogenesis. Prior to the stimulation of steroidogenesis it was observed that most of the tubulin present in these cells, as determined by indirect immunofluorescence, was in a 0.2-0.6 micrometers dia. granular form. When these cells were treated with ACTH and cAMP, respectively, it was observed that the granular form of tubulin was replaced by many organized microtubules. These granules were identified in the electron microscope using tubulin antibody/ferritin localization and appeared to be membrane-bound and identical to structures previously described as containing cholesterol. We have isolated these structures using cell homogenization and sucrose gradient centrifugation and analyzed the steroid composition by thin layer chromatography (94% cholesterol, 6% cholesterol ester). These granules also contained tubulin as determined by gel electrophoresis. In addition, they contained acid phosphatase as determined by their ability to hydrolize beta glycerolphosphate. We suggest that tubulin may be involved in the sequestering of cholesterol by preventing its transport to the mitochondria where conversion to pregnenolone takes place, and that steroidogenesis is increased when tubulin is dissociated from the cholesterol granules. PMID- 6273136 TI - A case with glucagonoma syndrome--endocrine and metabolic studies. AB - The results of clinical endocrine and metabolic studies on a 57-year-old female with surgically and autopsy verified glucagonoma syndrome were presented. All of the clinical manifestations of glucagonoma syndrome so far reported in the literature were noted but there was no evidence indicating the presence of multiple endocrine adenomatosis. The plasma IRG level was always more than 20 times above the normal, and the IRG response to insulin and tolbutamide injection was abnormal and the results of the other endocrinological studies revealed less remarkable features, if any. The surgically removed metastatic tumor of the liver contained an enormous amount of IRG and an appreciable amount of IRI, indicating that the elevated plasma IRG was mainly of tumor origin. These results clearly indicate that in glucagonoma there is some abnormality in glucagon release from the tumor. In addition to these findings, hypocalcemia, cardiac left ventricular hypertrophy and gastrointestinal dysfunction reportedly due to hyperglucagonemia were also seen in this patient. PMID- 6273137 TI - Inverse correlation between urinary 17-hydroxycorticosteroid (17-OHCS) excretion and thyrotropin-releasing hormone (TRH)-induced thyrotropin (TSH) response in disorders of adrenocorticotropic hormone (ACTH) and adrenal secretion. AB - A TRH test was performed in patients with Cushing's syndrome and isolated ACTH deficiency, and serum TSH was measured. A TRH-induced TSH showed low response in patients with Cushing's syndrome but showed excessive response in patients with isolated ACTH deficiency. However, in both diseases TRH-induced TSH response showed a tendency to normalize after the treatment. Urinary 17-OHCS excretion was measured in patients with Cushing's syndrome and isolated ACTH deficiency before and after the treatment. There was a definite inverse correlation between ;the logarithm for urinary 17-OHCS excretion and the peak TSH response to TRH. These results suggested that glucocorticoids regulate not only ACTH but also TSH secretion. PMID- 6273138 TI - Follow-up study on treatment in 27 patients with Cushing's disease: adrenalectomy, transsphenoidal adenomectomy and medical treatment. AB - 27 patients with Cushing's disease were treated over a period of 18 years at the Departments of Medicine and Surgery, Nagoya University School of Medicine and the following results were obtained. 1) Adrenalectomy. 21 of 27 patients with Cushing's disease underwent adrenalectomy. 19 patients had total bilateral adrenalectomy and 2 patients unilateral adrenalectomy. 4 patients died, the cause of death not being related directly to adrenalectomy. 9 of 15 bilaterally adrenalectomized patients had hyperpigmentation even though they had been given substitution therapy with cortisol 20-30 mg daily. They had elevated plasma ACTH levels, which were not completely suppressed by 2 mg of dexamethasone or 2.5 mg of bromocriptine per day. 2) Adenomectomy, 5 patients had adenomectomy via the transsphenoidal approach. 3 patients were cured but one of them has required postoperative substitution therapy with cortisol for hypopituitarism for one year until today. 2 of 5 adenomectomized patients had a recurrence of Cushing's syndrome after remission for 6-8 months. One of these recurrent cases has been subsequently treated successfully with bromocriptine, a dopaminergic drug. 3) Medical treatment. 2.5 mg per day of bromocriptine has been effective in 2 patients without a pituitary adenoma and ineffective in the other 4 patients with a pituitary adenoma. 24 mg per day of cyproheptadine, an antiserotoninergic drug was not effective in any of the 4 patients with a pituitary adenoma. PMID- 6273139 TI - Isopycnic density values for lysosomes and mitochondria in rat adrenal cortex. AB - Adrenocortical tissues of male adult Wistar rats were fractionated by isopycnic density gradient centrifugation. Fractions were analyzed for density, protein and marker enzymes for lysosomes and mitochondria with rat liver being used as a reference tissue for subcellular enzyme distribution. Both lysosomes and mitochondria of adrenal cortex showed unimodal distribution profiles of marker enzymes with their modal isopycnic density values at 1.165. This value was significantly lower than the corresponding ones for lysosomes and mitochondria in rat liver but was very close to those in porcine adrenal cortex. Modal isopycnic density as well as distribution profiles of marker enzymes for lysosomes and mitochondria remained unchanged 24 hr after 0.1 or 10 units of ACTH (Cortrosyn Z) administration. As in porcine adrenal cortex, lysosomes in rat adrenal cortex were characterized by a higher content of cathepsin D than those in rat liver. PMID- 6273140 TI - Casein kinase in cytosol of rat and mouse mammary epithelial cells isolated from histone kinase by MgCl2 treatment and influence of pregnancy and lactation on the enzyme activity. AB - Rat mammary glands contain cyclic AMP-independent casein kinase and cyclic AMP dependent histone kinase. The former was easily isolated from cyclic AMP dependent histone kinase by MgCl2 treatment. Mammary casein kinase was not activated by cyclic nucleotides, and Mg++ and ATP were required for activation. The specific activity of casein kinase in cytosol of rat mammary epithelial cells increased 2 to 3-fold during pregnancy and lactation. Cytosol of mouse mammary epithelial cells also contained cyclic AMP-independent casein kinase, and the activity of this enzyme was about three times that of the Golgi fraction. PMID- 6273141 TI - Adrenergic modulation of insulin and glucagon secretion from the isolated perfused rat pancreas. AB - In order to observe the effect of the adrenergic system on pancreatic glucagon secretion in the isolated perfused rat pancreas, phenylephrine, an alpha adrenergic agonist, and isoproterenol, a beta-adrenergic agonist, were added to the perfused solution. 1.2 microM phenylephrine suppressed glucagon secretion at 2.8 mM glucose, and it also decreased insulin secretion at 11.1 mM glucose. 240 nM isoproterenol enhanced glucagon secretion not only at 2.8 mM glucose, but also at 11.1 mM glucose, as well as insulin secretion at 11.1 mM. In order to study the role of intra-islet noradrenalin, phentolamine, an alpha-adrenergic antagonist, and propranolol, a beta-adrenergic antagonist, were infused with the perfused solution. 10 and 100 microM phentolamine caused an increase in insulin secretion, and 25 microM propranolol decreased insulin secretion, while they did not cause any change in glucagon secretion. From these results, it can be concluded that alpha-stimulation suppresses not only insulin but also glucagon secretion, while beta-stimulation stimulates glucagon secretion, as well as insulin secretion. Intra-islet catecholamine may have some effect on the B cell, whereas it seems to have no influence on the A cell. PMID- 6273142 TI - Effect of prolactin on the secretion of dehydroepiandrosterone (DHEA), its sulfate (DHEA-S), and cortisol by the human fetal adrenal in vitro. AB - The effect of prolactin on the secretions of dehydroepiandrosterone (DHEA) and its sulfate (DHEA-S) as well as that of cortisol were studied in vitro in order to investigate the possible regulatory role of prolactin on steroidogenesis of the human fetal adrenal at mid-gestational age. The addition of 0.5 microgram/ml of human prolactin to the incubation medium produced a significant (P less than 0.05) increase in DHEA, DHEA-S, and cortisol secretion. These results indicate that prolactin has a regulatory role in steroidogenesis in the human fetal adrenal at mid-gestation. PMID- 6273143 TI - Effect of gonadectomy and steroid treatment on the receptor-binding activity and immunoreactivity of serum and pituitary FSH in the adult rats of both sexes. AB - The receptor binding activity (R) and immunoreactivity (I) of serum and pituitary FSH were measured in normal, gonadectomized, and gonadal steroid-treated rats of both sexes employing radioreceptor assay and radioimmunoassay, respectively, and R/I ratios were analyzed. At 11 weeks of age, R/I ratios of serum and pituitary FSH were significantly higher in normal females than in males. From 11 to 13 weeks of age, the ratios of both FSH further increased in females, while in males, the ratio of serum FSH decreased and that of pituitary FSH increased. Orchiectomy caused an increase in the ratio of serum FSH, while ovariectomy caused a decrease, so the ratios in the gonadectomized animals of both sexes became almost equal, suggesting the secretion of a gonadectomy type FSH in these animals. Both testosterone propionate (TP) and estradiol benzoate (EB) treatments decreased the ratios of serum FSH in gonadectomized rats. However, TP and EB had an effect opposite on the ratio of pituitary FSH in these rats, i.e. TP caused a decrease, while EB caused an increase. This effect was more obvious in females than in males. These observations indicated that the presence and absence of the gonads, and the gonadal steroids influence not only the quantity but also the quality of FSH. The changes in the quality of the hormone are discussed in relation to its multiplicity. PMID- 6273144 TI - Hypertension, hypokalemia and hypoaldosteronism with suppressed renin: a clinical study of a patient with Liddle's syndrome. AB - A 24-yr-old woman with hypertension, hypokalemic alkalosis, low plasma renin and hypoaldosteronism was studied. Plasma aldosterone, renin and potassium returned to normal and blood pressure fell after sodium restriction or the administration of triamterene. Thiazide therapy also normalized her blood pressure while dexamethasone, spironolactone and furosemide did not improve her symptoms. Plasma aldosterone levels were low and responded poorly to a short term ACTH injection, but responded well to the maximal adrenal stimulation by ACTH-Z. Plasma levels of cortisol, corticosterone and deoxycorticosterone were within the normal range. Adrenal scintigram with 131I-adosterol and abdominal computed axial tomography did not reveal the presence of a sizeable adrenal tumor. In addition, the urinary kallikrein excretion was low after sodium restriction and showed no response to saline infusion. These findings suggest that the excessive secretion of unusual mineralocorticoids may not exist in this case. From these observations and the results of the therapeutic responses to the diuretic agents, we conclude that the primary cause of the disorder of this patient seems to be a renal defect in the distal tubule in handling sodium and potassium which is similar to that in Liddle's syndrome. PMID- 6273145 TI - Species specificity of radioreceptor assay and radioimmunoassay for rat FSH. AB - Species specificity of the radioreceptor assay (RRA) for rat FSH, in which pregnant mare serum gonadotropin (PMSG)-treated immature rat ovary was employed as the receptor, was compared with that of NIAMDD rat FSH radioimmunoassay (RIA). In the RIA system, pituitary preparations from mammals only showed significant crossreaction. Their inhibition curves, however, were not always parallel to the standard curve. On the other hand, in the RRA system, the pituitary preparations from mammals, avians, lizard and amphibians competitively inhibited the binding of radioactive rat FSH to the ovarian receptor. Only the pituitary preparation from dog salmon failed to show any crossreaction in the RRA system. These results indicated that this RRA system would be useful for the measurement of FSH or gonadotropins of the pituitaries from mammals to amphibians. PMID- 6273146 TI - The neuroendocrine regulation and function of growth hormone and prolactin in the mammalian fetus. PMID- 6273147 TI - Extrapituitary actions of gonadotropin-releasing hormone. PMID- 6273149 TI - GABA in cerebrospinal fluid of children with febrile convulsions. AB - In 23 children with febrile convulsions the concentration of gamma-aminobutyric acid (GABA) in lumbar cerebrospinal fluid (CSF) was measured by a radioreceptor assay. The mean CSF GABA concentration of 134 (range, 73-294) pmoles/ml was significantly lower than that of 16 seizure-free children serving as controls, who had 210 (range, 117-475) pmoles/ml. The reduction in CSF GABA levels in patients with febrile convulsions was not reflected in plasma GABA concentrations. These data provide further evidence that impairment of GABA neurotransmission may contribute to an increased seizure propensity. PMID- 6273148 TI - Isolated aldosterone deficiency in man: acquired and inborn errors in the biosynthesis or action of aldosterone. PMID- 6273150 TI - Allosteric properties of rat lung phosphofructokinase. AB - Rat lung phosphofructokinase is purified 250-fold to a specific activity of about 10 by using ATP-sepharose affinity chromatography. The enzyme is activated by cyclic AMP, 5'-AMP, ADP, Pi, NH4+ and K+ ions. Depending upon the concentration of these effectors, the enzyme can exist in several interconvertible forms, differing widely in their affinity for fructose-6-P. These activators also overcome the inhibition of the enzyme by ATP and citrate, thus increasing the glycolytic rate in lung during hypoxia. Unlike the enzyme from other sources, the lung phosphofructokinase is not inhibited by cyclic GMP or phosphoenolpyruvate. The enzyme is very sensitive to inactivation by trypsin and this inactivation is completely reversed by assaying the proteolyzed enzyme in presence of its activators. PMID- 6273151 TI - Correlation between protein phosphorylation and progesterone synthesis in bovine luteal cells stimulated by lutropin. PMID- 6273152 TI - The interaction of the high-density lipoprotein with cultured cells of bovine vascular endothelium. PMID- 6273153 TI - Up-regulation in vascular endothelial cells of binding sites of high density lipoprotein induced by 25-hydroxycholesterol. AB - Exposure of bovine vascular endothelial cell cultures to 25-hydroxycholesterol (50--100 microgram/ml) result in a 5--10-fold increase in cell surface binding sites of high density lipoprotein (HDL). This increase in HDL-binding sites was dependent on time and temperature. After a 48-h exposure to the oxygenated sterol, a maximal increase in HDL binding could be observed, and newly binding sites disappeared rapidly once 25-hydroxycholesterol was removed from the medium. No increase in HDL-binding sites was observed when cells were maintained at 4 degrees C. In contrast, cultures maintained at 37 degrees C did show an increase in HDL-binding sites when exposed to 25-hydroxycholesterol. Since simultaneous exposure of the cells to 25-hydroxycholesterol and cycloheximide resulted in an inhibition of HDL binding to the cells, it is suggested that de novo synthesis of HDL-binding sites is induced by 25-hydroxycholesterol. When the abilities of HDL2 and HDL3 to bind to newly synthesized HDL-binding sites were compared, HDL3 was found to bind more efficiently than HDL2. It is therefore unlikely that apoprotein E plays a major role in the binding of HDL to newly synthesized HDL binding sites. When the properties of newly synthesized HDL-binding sites were analyzed, they were found to have a high affinity for HDL, since half-maximal binding was reached at a concentration as low as 5 microgram HDL protein/ml, and were saturable. Such HDL-binding sites had a relaxed specificity, since they were capable of binding low density liprotein (LDL). However, when LDL bound to newly synthesized HDL binding sites, it was no longer internalized, as reflected by a 90% reduction of LDL degradation, and instead of being cytotoxic it became mitogenic. PMID- 6273154 TI - The cdc 22 mutation by Schizosaccharomyces pombe is a temperature-sensitive defect in nucleoside diphosphokinase. AB - A number of temperature-sensitive cdc- mutants of Schizosaccharomyces pombe that are affected in DNA replication, were screened for the absence of deoxynucleoside triphosphate(s) when blocked at their restrictive temperature. The preliminary screening simply involved analysis of perchloric acid-soluble cell extracts by two-dimensional thin-layer chromatography on poly(ethyleneimine)-impregnated cellulose. One mutant strain, cdc 22-M45, was found which apparently lacked dTTP. Pulse-labelling of intracellular nucleotides revealed that not only did dTTP become depleted, but that dTDP accumulated when this mutant was blocked by a temperature shift-up, indicating a defective nucleoside diphosphokinase. Nucleoside diphosphokinase from cdc 22-M45 was less active than that from wild type strain 972 when assayed at high temperatures. The nucleoside diphosphokinase of the mutant also has an altered Km for dTDP at both permissive (25 degrees C), and at the restrictive (36.8 degrees C) temperatures. At the restrictive temperature the Km for dTDP of the mutant enzyme is more than 11-times greater than that of the wild type. Characterisation of the biochemical basis of the defect in this cdc- mutant has shown that in S. pombe, despite its having an apparently complex system of genetic control over progression through S-phase, one factor at least is merely availability of a nucleoside triphosphate precursor to DNA synthesis. PMID- 6273155 TI - Subunit stoichiometry of cytochrome c oxidase of bovine heart. AB - Cytochrome c oxidase from bovine heart was dissociated into its protein subunits by sodium dodecylsulphate, the subunits were separated on a preparative scale by sodium dodecylsulphate gel permeation chromatography. The subunits elute upon gel chromatography in order of decreasing apparent relative molecular mass (I, 40000; II, 26000; III, 21000; IV, 17000; V and VI, 12000; VII, 10000 and VIII, 6000). The very hydrophobic subunits I and III tend to form small aggregates both in the presence and absence of sodium dodecylsulphate. The molar ratio of the subunits was determined by two methods: firstly by quantitative amino acid analysis of each subunit peak, and secondly from the absorbance of each subunit at 280 nm caused by tryptophan and tyrosine. We conclude that the subunits I to VI are present in 1 : 1 ratio; our fraction VII contains two stoichiometric polypeptides which may or may not be identical. Fraction VIII contains enough protein for four stoichiometric chains which may belong to three different types. The 12 stoichiometric chains add up to a Mr of about 170000 if the sizes of I and III are 40000 and 21000, respectively. After correction for the presence of aggregates, subunits I and III appear to be present in more than 1 : 1 stoichiometric amounts with respect to other subunits, which probably means that subunits I and III are considerably larger than hitherto assumed. This is in line with recently published mtDNA sequence work [Anderson, S. et al. (1981) Nature, 290, 457--465]. PMID- 6273156 TI - The specificity of the sodium channel for monovalent cations. PMID- 6273157 TI - Purification of glycogen synthase kinase 3 from rabbit skeletal muscle. Copurification with the activating factor (FA) of the (Mg-ATP) dependent protein phosphatase. PMID- 6273158 TI - Is increased basal lipolysis in adipose tissue of fasted-refed rats related to cyclic-AMP-dependent mechanisms? PMID- 6273159 TI - Arginyl-tRNA synthetase from Baker's yeast. Order of substrate addition and action of ATP analogs in the aminoacylation reaction; influence of pyrophosphate on the catalytic mechanism. AB - The order of substrate addition to arginyl-tRNA synthetase from baker's yeast has been investigated by bisubstrate kinetics, product inhibition and inhibition by three different inhibiting ATP analogs, the 6-N-benzyl, 8-bromo and 3'-deoxy derivatives of ATP, each acting competitively with respect to one of the substrates. The kinetic patterns are consistent with a random ter-ter mechanism, an addition of the three substrates and release of the products in random order. The different inhibitors are bound to different enzyme . substrate complexes of the reaction sequence. Addition of inorganic pyrophosphatase changes the inhibition patterns and addition of methylenediphosphonate as pyrophosphate analog abolishes the effect of pyrophosphatase, showing that the concentration of pyrophosphate is determinant for the mechanism of catalysis. PMID- 6273160 TI - Solvent and temperature effects on crambin, a hydrophobic protein, as investigated by proton magnetic resonance. PMID- 6273161 TI - Molecular properties of chloroplastic thioredoxin f and the photoregulation of the activity of fructose 1,6-bisphosphatase. PMID- 6273162 TI - Purification and characterization of Mg2+-dependent glycogen synthase phosphatase (phosphoprotein phosphatase IA) from rat liver. AB - Phosphoprotein phosphatase IA, which represents the major glycogen synthase phosphatase activity in rat liver cytosol, has been purified to apparent homogeneity by chromatography on DEAE-cellulose, histone - Sepharose-4B and Sephadex G-100. The molecular weight of the purified enzyme was 40 000 by gel filtration and 48 000 by sodium dodecyl sulfate gel electrophoresis, Phosphatase IA is therefore a monomeric protein. When treated with 80% ethanol at room temperature, phosphatase IA underwent an inactivation which was totally prevented by 2 mM MgCl2. Catalytically, phosphatase IA has a preference for glycogen synthase D compared with phosphatases IB and II and obligatorily requires Mg2+ or Mn2+ for activity. Maximum activity was attained at 5 mM MgCl2. Since Mg2+ does not activate other phosphoprotein phosphatases in rat liver cytosol, we propose the term 'Mg2+-dependent glycogen synthase phosphatase' for phosphatase IA. PMID- 6273163 TI - Structural studies of the non-histone chromosomal proteins HMG-T and H6 from trout testis. AB - Trout testis contains two proteins of the high mobility group H6 and HMG-T that have been implicated in the structure of active chromatin. Protein H6 was studied by high resolution proton NMR and by circular dichroism and showed no evidence of secondary or tertiary structure formation in free solution. At low ionic strength protein H6 binds to DNA by a weak interaction in the N-terminal region between residues 10 and 35. Proteins H6 therefore behaves structurally like the homologous calf-thymus high-mobility-group proteins HMG-14 and HMG-17. Salt addition to solutions of protein HMG-T results in secondary and tertiary structure formation that is accompanied by aggregation. Circular dichroism shows that the helical content of protein HMG-T (approximately equal to 9%) is very much less than that of the homologous calf thymus proteins HMG-1 and HMG-2. At low ionic strength protein HMG-T precipitates DNA due to the formation of large scale aggregates that disperse only when the protein is released at approximately equal to 0.35 M NaCl. The NMR spectrum of the aggregated state does not show the presence of a large number of free acidic residues, in contrast to spectra of soluble complexes of HMG-1 and DNA at the same ionic strengths. It is concluded that HMG-T lacks the highly acidic domain found in HMG-1 (and HMG-2) that remains free from DNA under these conditions. It is concluded that the entire chain of HMG-T binds to DNA. There are therefore major structural differences between HMG T and the homologous calf thymus proteins. PMID- 6273164 TI - An electron-spin-resonance study on the redox-active centers of the 4 methoxybenzoate monooxygenase from Pseudomonas putida. PMID- 6273165 TI - The essential carboxyl group in restriction endonuclease EcoRI. AB - We have carried out studies on type II restriction endonuclease EcoRI, which cleaves the DNA sequence 5'd(-G-A-A-T-T-C-)3', as indicated. The active form of the enzyme consists of two subunits, each 31063 molecular weight. A water-soluble reagent, 1-cyclohexyl-3-(2-morpholinoethyl)carbodiimide metho-p-sulphonate, which reacts with carboxyl groups and also with tyrosine and cysteine residues, has been found to inactivate this enzyme. Results are presented which show the following. (1) This specific inactivation is not due to modification of tyrosine or cysteine residues. (2) There is one carboxyl group per subunit which, when modified with carbodiimide, inactivates the enzyme. (3) phi X174 DNA (which does not contain EcoRI sites) partially protects the enzyme from the carbodiimide; protection is unaffected by the additional presence of Mg2+, but significantly greater with Co2+ and phi X174 DNA. PMID- 6273166 TI - Induction of polymerization of purified tubulin by sulfonate buffers. Marked differences between 4-morpholineethanesulfonate (Mes) and 1,4 piperazineethanesulfonate (Pipes). AB - Interactions of both purified tubulin and microtubule protein (tubulin plus associated proteins) with two commonly used sulfonate buffers were examined. 1,4 Piperazineethanesulfonate (Pipes) and 4-morpholineethanesulfonate (Mes) at high concentrations induce the polymerization of purified tubulin in reactions requiring only buffer, tubulin and GTP. While both reactions were temperature dependent, cold-reversible and inhibited by GDP, colchicine or Ca2+, there were significant differences between them. Substantially lower tubulin and buffer concentrations were required for Pipes-induced polymerization; and turbidity was much more intense in the Pipes-induced than in the Mes-induced reaction at the same protein concentration. Electron microscopy demonstrated that for the most part typical smooth-walled microtubules were formed in Mes, while aberrant forms were the predominant structures formed in Pipes. When the polymerization of microtubule protein was examined as a function of buffer concentration, biphasic patterns were observed with both Pipes and Mes: polymerization occurred at both low and high, but not intermediate, buffer concentrations. The turbidity observed at high concentrations of Pipes greatly exceeded that at low concentrations. With Mes, equivalent turbidity developed at both high and low buffer concentrations. Although associated proteins copolymerized with tubulin at low buffer concentrations, they were excluded from the polymerized material at high buffer concentrations. Pipes and Mes were compared to sodium phosphate, Tris/HCl and imidazole/HCl buffers at 0.1 M in several polymerization systems using both purified tubulin and microtubule protein. The sulfonate buffers were invariably associated with more vigorous reactions than the other buffers. PMID- 6273167 TI - A new principle of regulation of enzymic activity. Activation and regulation of human polymorphonuclear leukocyte collagenase via disulfide-thiol exchange as catalysed by the glutathione cycle in a peroxidase-coupled reaction to glucose metabolism. PMID- 6273168 TI - Formation of leukotrienes E3, E4 and E5 in rat basophilic leukemia cells. AB - Rat basophilic leukemia (RBL-1) cells incubated with ionophore A23187 and 5,8,11 eicosatrienoic acid produced three slow-reacting substances identified as leukotrienes C3, D3 and E3 by spectroscopic, chromatographic and enzymatic methods. 5,8,11,14,17-Eicosapentaenoic acid was similarly converted by RBL-1 cells to leukotrienes C5, D5. and E5. Leukotrienes C4, D4 and E4 were also formed in these experiments from endogenous arachidonic acid. Time-course studies, incubations with 3H-labeled leukotriene C3 and effects of acivicin [L-(alpha S, 5S)-alpha-amino-3-chloro-4,5-dihydro-5-isoxazoleacetic acid; a gamma-glutamyl transpeptidase inhibitor] indicated that leukotrienes C and D are intermediates in the formation of leukotrienes E. L-Cysteine enhanced the conversion of leukotriene C3 to leukotriene D3 and inhibited further degradation of leukotriene D3 to leukotriene E3. PMID- 6273169 TI - The catabolism of phosphatidylinisitol by an EDTA-insensitive phospholipase A1 and calcium-dependent phosphatidylinositol phosphodiesterase in rat brain. AB - 1. A rat brain supernatant and microsomal fraction contained a phospholipase A1 enzyme which hydrolysed phosphatidylinositol at pH 8 in the absence of calcium. Triolein and phosphatidylcholine were not attacked under the same incubation conditions. 2. No evidence could be obtained for a phospholipase A2 in the microsomal preparation, and in the presence of Ca2+ the release of fatty acid observed was due to phosphatidylinositol phosphodiesterase followed by diacylglycerol lipase action. 3. Brain phosphatidylinositol phosphodiesterase showed extensive activity in the alkaline range (7-8.5) as well as at pH 5-5.5. The activity at higher pH values required higher calcium concentrations and disappeared on purification of the soluble enzyme by ammonium sulphate fractionation. 4. In general the ratio between inositol 1,2-(cyclic)phosphate and inositol 1-phosphate produced by phosphodiesterase action decreased with increasing pH. PMID- 6273170 TI - Endoplasmic reticulum nuclease. Purification and specificity. AB - An endonuclease, which was originally identified for its RNA polymerase inhibitory activity, was isolated from rat liver endoplasmic reticulum. The enzyme yields on gel chromatography four active fractions of different molecular weights (Mr 5.3 X 10(4), 9 X 10(4), 1.55 X 10(5) and Sephacryl S-200 fraction at V0). Each fraction contains polypeptide chains which give a single band on sodium dodecylsulphate electrophoresis (Mr 5.4 X 10(4). This indicates that the enzyme is an oligomeric protein and each of its subunits exhibits the same or very similar molecular weights. Deoxyribonucleoside and ribonucleoside triphosphates can bind to the endoplasmic reticulum nuclease. Binding is enhanced in the presence of divalent cations particularly Mg2+. The enzyme exhibits mainly RNase activity but can also degrade denatured DNA and DNA . RNA hybrids which contain breaks in one of the two strands. Poly(A) and mainly poly(U) are most susceptible to its nucleolytic activity whereas poly(C) is completely resistant. PMID- 6273171 TI - Quantification of left ventricular volume in gated equilibrium radioventriculography. PMID- 6273172 TI - An evaluation of 99mTc pertechnetate scanning for the detection of coeliac disease and Crohn's disease. AB - To investigate the use of abdominal scintiscanning in the detection of small bowel pathology, the accumulation of pertechnetate (99mTc), following intravenous injection, has been studied in 21 patients with coeliac disease, 13 patients with Crohn's disease and in 83 control subjects without known small bowel disease. Although a trend for a greater accumulation of 99m Tc was noted in patients with coeliac disease and Crohn's disease compared with controls (P less than 0.025) there was a large overlap in individual studies. Under the conditions of this study the accumulation of 99mTc by the small bowel did not provide a reliable diagnostic test for coeliac disease or Crohn's disease of the small intestine. The accumulation of 99mTc pertechnetate by normal small bowel suggests that scintiscanning with this radiopharmaceutical does not provide a consistently reliable method for the detection of small bowel pathology. PMID- 6273173 TI - The biological fate of sulphur colloid. AB - The in-vivo behaviour of sulphur colloid has been investigated using colloids labelled with 35S as well as 99m Tc. The rates of clearance of 35S and 99m Tc from the blood, the rates of accumulation in liver and bone and the distribution of the two radioisotopes in various organs are all markedly different. The results demonstrate that although technetium is rapidly removed from the blood stream and primarily accumulated in the liver the colloid particles themselves are broken down in vivo with the release of sulphur. PMID- 6273174 TI - Acute anticoagulant-induced compressive lumbar plexus neuropathy. A clinico pathological study. PMID- 6273175 TI - Absence of antigens related to murine mammary tumour virus polypeptides in rat mammary tumours. PMID- 6273176 TI - Hodgkin's disease following infectious mononucleosis. A case report. PMID- 6273177 TI - Herpes simplex virus and human cancer. I. Relationship between human cervical tumours and Herpes simplex type 2. PMID- 6273178 TI - Herpes simplex virus and human cancer. II. Search for relationship between labial tumours and Herpes simplex type 1. PMID- 6273179 TI - Use of labetalol in hypertensive patients during discontinuation of clonidine therapy. AB - Eleven hypertensive patients in whom clonidine therapy had to be discontinued, were treated prophylactically with labetalol, in order to avoid a possible hypertensive crisis. Most of the known side effects, which are consistent with the withdrawal phenomenon were observed, e.g. tremor, insomnia and apprehension, but headaches and flushing did not occur. Blood pressure levels remained unchanged, despite up to a 20-fold increase in plasma catecholamines. The lack of change in serial measurements of plasma cyclic AMP level appears to indicate that adequate adrenergic blockade was induced by labetalol. Since labetalol is a potent anti-hypertensive drug, and is also effective in avoiding a possible hypertensive crisis due to withdrawal of clonidine, we propose to use it as the drug of choice whenever discontinuation of clonidine therapy is indicated. PMID- 6273180 TI - A comparison of the bioavailability and potency of dexamethasone phosphate and sulphate in man. AB - The metabolic fate and ACTH-suppressant activity of two injectable dexamethasone esters, 21-phosphate and 21-sulphate, were studied in healthy men. After i.v. injection of 20mg free steroid alcohol, dexamethasone phosphate was efficiently hydrolyzed to free dexamethasone, reaching its peak plasma concentration within 5 min. About 9% of the administered dose appeared in the urine as free dexamethasone. By contrast, virtually no free dexamethasone was found in plasma and urine after injection of dexamethasone sulphate. Pharmacokinetic analysis showed that dexamethasone sulphate had a shorter plasma half-life and a higher metabolic clearance rate than free dexamethasone. A larger fraction (60%) of dexamethasone sulphate was rapidly excreted unmetabolized in urine. The plasma cortisol level was significantly suppressed for more than 24h after dexamethasone phosphate, while the plasma cortisol profile after dexamethasone sulphate merely showed physiological circadian variations. When the steroid esters were injected after pretreatment with metyrapone, a definite suppression of plasma ACTH was noted after dexamethasone phosphate, but again, dexamethasone sulphate was ineffective. These results cast serious doubt on the clinical value of dexamethasone sulphate as an injectable glucocorticoid, and critical reevaluation of this preparation is needed. PMID- 6273181 TI - Purification and characterization of an immunosuppressive factor from ovarian cancer ascites fluid. AB - A nonspecific immunosuppressive factor present in malignant (ovarian carcinoma) ascites fluid has been purified by acid extraction from a high molecular weight (greater than 20000) complex followed by preparative isoelectric focusing on Bio lyte media. It is an acidic protein (pI = 3.6) of mol. wt. 50000 to 52000 as estimated by gel filtration and composed of subunits of 25000 to 26000 estimated by sodium dodecyl sulfate polyacrylamide gel electrophoresis, under reducing conditions. It inhibits the phytohemagglutinin-dependent mitogenic response of normal peripheral blood lymphocytes by 50% at 2 microgram/ml concentrations in vitro and suppresses 80% of the plaque-forming cell response to sheep erythrocytes at 100 microgram per mouse in vivo. Its chemical identity with any of the known plasma proteins has not been established. Its failure to stain with periodic acid Schiff's reagent indicates its minimal content of carbohydrates. It is susceptible to tryptic and pronase digestion but insensitive to deoxyribonuclease and ribonuclease digestion. A smaller suppressive factor identified in the same fluid appears to be a lymphotoxin; it differs from the acid-extracted nonspecific suppressive factor in its lack of susceptibility to trypsin. PMID- 6273182 TI - The killing of newborn larvae of Trichinella spiralis by eosinophil peroxidase in vitro. AB - Helminth infections in mammals are characterized by a high level of eosinophils in parasitized tissues and blood, and it has recently been suggested that these cells have a direct parasiticidal effect. Newborn larvae of Trichinella spiralis can be killed within 20 min by incubation at room temperature in a cell-free system, including purified human eosinophil peroxidase (EPO), H2O2 and chloride at pH 5.5. Killing was measured by microscopic observation of the larvae. The larvicidal effect was dependent on each component of the EPO/H2O2/Cl- system and could be prevented by using SO4(2-) instead of Cl-. Killing was totally inhibited by sodium azide and catalase, and substantially by bovine serum albumin, a protein that is an effective scavenger for HOCl. Since larvae could also be killed directly by HOCl under these conditions and EPO is able to oxidize Cl- to hypochlorous acid, it is very likely that the larvicidal effect of the EPO system is due to formation of hypochlorous acid. It is proposed that in vivo, the combination of EPO, which is exocytosed onto the surface of the parasite, and H2O2, which is generated by stimulated eosinophils, is responsible for the larvicidal effect. PMID- 6273183 TI - Potentiation by spiperone and other butyrophenones of fluid secretion by isolated salivary glands of ixodid ticks. AB - Isolated salivary glands from the ixodid tick, Amblyomma hebraeum Koch are stimulated to secrete fluid when exposed to dopamine (DA), the maximum response occurring at 10(-6) M. Spiperone, and a number of other butyrophenone derivatives, although lacking intrinsic activity, are able to potentiate the secretion elicited by supramaximal concentrations of DA; this potentiation by spiperone is evident at concentrations in the femtomolar range. Tranylcypromine, a potent, competitive inhibitor of monoamine oxidase (MAO) in tick salivary gland homogenates, has both intrinsic activity and potentiates DA-induced salivation. The fact that spiperone potentiates ergometrine-induced salivation that the prime mechanism of the butyrophenone effect is not by inhibiting catecholamine catabolism. The results also suggest that the receptor for DA and that for butyrophenones are distinct sites. Droperidol, benperidol and bromperidol, all potent neuroleptic drugs, failed (at 10(-9) M) to potentiate salivation. By contrast, R951, R27275 and R1187 (all at 10(-9) M) were very effective potentiators on the salivary gland system, despite the fact that they lack the basic structural requirements for neuroleptic activity. These results suggest that the butyrophenone site in tick salivary glands is different from butyrophenone binding sites in mammalian CNS. PMID- 6273184 TI - Ionic mechanism of diphenylhydantoin action on glucose-induced insulin release. AB - Diphenylhydantoin inhibits glucose-stimulated insulin release. The mode of action of diphenylhydantoin was investigated by characterizing its effect on 86Rb and 45Ca fluxes in isolated pancreatic islets. The inhibition of glucose-stimulated insulin release by diphenylhydantoin was apparently not attributable to activation of a Na+ + K+ATPase as diphenylhydantoin failed to affect 86Rb net uptake, at least in glucose-stimulated islets. Diphenylhydantoin decreased 45Ca net uptake by the islets, an effect possibly due to inhibition of Ca2+ entry into the islet cells. Diphenylhydantoin indeed markedly inhibited the glucose-induced increase in 45Ca outflow and decreased the process of 40Ca-45 exchange evoked by a rise in extracellular Ca2+ concentration. Diphenylhydantoin failed to affect the inhibitory action of glucose upon 45Ca outflow whether in the presence or absence of extracellular Ca2+, and did not impair the antimycin A-induced release of 45Ca from intracellular organelles. These findings suggest that the inhibitory effect of diphenylhydantoin upon glucose-stimulated insulin release is attributable mainly to a blockade of Ca2+ inflow into the beta-cell. PMID- 6273185 TI - Peripheral benzodiazepine binding sites in heart and their interaction with dipyridamole. AB - Specific binding of [3H]diazepam to rat and guinea-pig heart tissue has been found. These binding sites are similar to those in lung, liver, kidney and mast cells but differ from those in the brain. Dipyridamole interacts with this receptor and is several fold more potent as a displacer of [3H]diazepam bound to heart preparations than to brain membranes. Specific binding per unit protein in the left ventricle, right ventricle and interventricular septum is significantly higher than in the left and right atrium. PMID- 6273186 TI - [125I]BE 2254, a new high affinity radioligand for alpha 1-adrenoceptors. AB - [125I]BE 2254 (IBE 2254), a new iodinated radioligand of high specific radioactivity (2175 Ci/mmol) was developed and used to characterize alpha 1 adrenoceptors in rat cerebral cortex membranes. IBE 2254 possessed a high affinity, KD 78 +/- 14 pM with a Bmax of 210 +/- 26 fmol/mg and 90% specific binding at KD value. alpha-Adrenergic antagonists competed for IBE 2254 binding in the following order: prazosin greater than or equal to WB 4101 greater than phentolamine greater than corynanthine greater than yohimbine greater than rauwolscine, a strong indication that IBE 2254 binds to alpha 1-adrenoceptors. IBE 2254 appears to be a very useful tool for studying alpha 1-adrenoceptors. PMID- 6273187 TI - On the opioid nature of phencyclidine and its 3-hydroxy derivative. AB - Phencyclidine (PCP) and its 3-hydroxy derivative (PCP-3-OH) caused a dose dependent, naloxone reversible inhibition of the response of the guinea pig ileum to electrical stimulation. Unlike PCP, PCP-3-OH exerted an opioid antagonistic effect in the mouse vas deferens bioassay. Whereas both compounds displayed a high affinity in displacing [3H]SKF-10047 binding to rat brain membranes, PCP-3 OH displayed a high affinity to [3H]morphine receptors also. The mediation of alpha- and mu-receptors in the opioid effects of these drugs is discussed. PMID- 6273188 TI - Rabbit vas deferens: a specific bioassay for opioid kappa-receptor agonists. PMID- 6273189 TI - Calcium-mediated transduction of the hormonal message in meiosis reinitiation of starfish oocytes: modulation following injection of cholera toxin and cAMP dependent protein kinase. PMID- 6273190 TI - Hormone-induced phosphorylation of a 16000 Dalton polypeptide following meiosis reinitiation in starfish oocytes. PMID- 6273192 TI - Vimentin-derived proteins: differences between normal human fibroblasts and transformed human cells. PMID- 6273191 TI - Hormonal regulation of DNA synthesis in primary cultures of adult rat hepatocytes: action of glucagon. PMID- 6273193 TI - cAMP uncouples DNA synthesis and cell division in Tetrahymena. PMID- 6273194 TI - Changes in cyclic nucleotide levels during erythroid differentiation in Friend leukemia cells. PMID- 6273195 TI - 1-beta-D-arabinofuranosylcytosine stimulates thymidine incorporation into the DNA of contact-inhibited cells. PMID- 6273196 TI - Inactivation by chloroquine of alpha-galactosidase in cultured human skin fibroblasts. PMID- 6273197 TI - Plasma membrane-mediated effects of extracellular pH on the growth of neuroblastoma cells. PMID- 6273198 TI - Immunocytochemical localization of a calcium-activated protease in skeletal muscle cells. PMID- 6273199 TI - Genetic determinants of virus susceptibility: epidemiologic implications of murine models. PMID- 6273200 TI - Feline leukemia virus-induced erythroid aplasia: in vitro hemopoietic culture studies. AB - Colony forming unit (CFU) assays were developed for feline granulocyte-macrophage (CFUGM), early erythroid (day 2 CFUE), and late erythroid (day 7 CFUE) colonies in methylcellulose medium. Feline CFUGM and both day 2 and day 7 CFUE were enhanced by feline macrophage conditioned medium and late CFUE often were intimately associated with macrophages. Kittens were inoculated with the Kawakami Theilen (KT) strain of feline leukemia virus (FeLV) and sequential changes in marrow CFU determined. Erythroid aplasia, characterized by progressive non regenerative anemia, lymphopenia, and a profound decrease in early and late CFUE but not CFUGM was induced by 3 to 5 weeks after FeLV-KT inoculation. The susceptibility of kittens to FeVL-induced erythroid aplasia was strongly age related; neonatal kittens were most sensitive and substantial natural resistance developed by 4 weeks of age. The results demonstrate that FeLV-KT infection induced a rapid and selective suppression of erythroid progenitor cells and represents a suitable model of experimentally-induced acquired erythroid aplasia. PMID- 6273201 TI - The diagnosis of sarcoidosis. PMID- 6273202 TI - Significance of serum angiotensin converting enzyme and gallium scan in noninvasive diagnosis of sarcoidosis. AB - Serum angiotensin converting enzyme (SACE) assays, chest roentgenograms and gallium scans were obtained in 28 patients with active sarcoidosis, two patients with resolved sarcoidosis and 43 nonsarcoid patients. In patients with active sarcoidosis, 23 had elevated SACE activity and 26 had diffuse uptake of gallium in the lung parenchyma. Normal SACE and gallium scans were found in patients with resolved sarcoidosis. None of the nonsarcoid patients had a combination of elevated SACE activity and diffuse pulmonary parenchymal uptake of gallium, although some had elevated SACE activity and abnormal gallium scans. These results suggest that combination of elevated SACE activity and diffuse accumulation of gallium in the lung parenchyma are suggestive of sarcoidosis. However, in view of the reports of abnormal gallium accumulation, and occasional elevation of SACE activity in silicosis, asbestosis and miliary tuberculosis, it is still necessary to require histological evidence of noncaseating granuloma to confirm the diagnosis of sarcoidosis. PMID- 6273203 TI - Long-term treatment with corticosteroids and ACTH in asthmatic children. I. A pulmonary function study with re-investigation after 5 years. AB - Twenty-one children aged 6-17 years with severe bronchial asthma and on long-term treatment with corticosteroids and/or ACTH, were studied when they were as symptom-free as possible. They were found to have increased lung volumes and hyperinflation with a high end-expiratory level. The forced expiratory 1-s volume (FEV1) was within +/- 2 SD for healthy children, but the FEV% was significantly decreased, as a sign of sub-clinical bronchial obstruction. Arterial oxygen tensions was within normal limits, except in one patient with clinical signs of bronchial obstruction. On re-examination 5 years later, hyperinflation and sub clinical obstruction were even more pronounced, despite the fact that the majority of the patients had improved clinically. PMID- 6273205 TI - Congenital cytomegalovirus effects on postnatal neurological development of squirrel monkey (Saimiri sciureus) offspring. PMID- 6273204 TI - The vibrissal pad as a source of sensory information for the oculomotor system of the cat. AB - Responses from lateral rectus, medial rectus and retractor bulbi nerves were obtained following electrical stimulation of the vibrissal pad of the cat. Discharges in afferent fibres dissected from the infraorbital nerve were recorded during movements of the vibrissae and following electrical stimulation of the vibrissal pad. Both stimuli activated the same population of A alpha fibers. Intracellular records were obtained from lateral rectus motoneurones identified antidromically in the principal abducens nucleus and from retractor bulbi motoneurones similarly identified in the accessory abducens nucleus. EPSPs (3 mV) were recorded in lateral rectus motoneurones following electrical stimulation of the ipsilateral vibrissal pad at a latency of 3.5 ms. Large-amplitude disynaptic EPSPs (15 mV) were recorded in retractor bulbi motoneurones following the same vibrissal stimulation. The synaptic excitation evoked in both lateral rectus and retractor bulbi motoneurones through stimulation of the ipsilateral vibrissal pad induced an early retraction followed by an abduction of the eye ball. The hypothesis is that the vibrissal message might complement other sensory modalities in the generation of patterned eye movements. PMID- 6273206 TI - Significance of cortical-amygdalar-hypothalamic connections in retention of conditional taste aversion in rats. PMID- 6273207 TI - Tumorigenic activity of cloned polyoma virus DNA in newborn rats. PMID- 6273208 TI - Interactions of delta 1-tetrahydrocannabinol with cannabinol and cannabidiol following oral administration in man. Assay of cannabinol and cannabidiol by mass fragmentography. PMID- 6273209 TI - Effect of suloctidil on blood viscosity in healthy volunteers after forearm occlusion. PMID- 6273210 TI - Lack of beneficial effects of wheat bran cereals on cholesterol balance in swine. PMID- 6273211 TI - [Neurochemical aspects of the pharmacology of GABAergic substances]. PMID- 6273212 TI - [Comparative study of the effect of lithium chloride and lithium oxybutyrate on carbohydrate metabolism in rabbits and rats]. PMID- 6273213 TI - [Effect of diethyldithiocarbamate on the activity of cellular antioxidative enzymes]. PMID- 6273214 TI - [Research on a substance with antiviral activity. XXI. Preparation and antiherpes activity of 2-acylamino-3,5-dichloropyridine]. PMID- 6273215 TI - A relationship between thiols and the superoxide ion. PMID- 6273216 TI - Cyclic-AMP activates and calcium inhibits protein kinase activity in avian parathyroid glands. PMID- 6273217 TI - Resonance Raman and surface-enhanced resonance Raman studies of cytochrome cd1. PMID- 6273218 TI - Preferential association of uracil-DNA glycosylase activity with replicating SV40 minichromosomes. PMID- 6273219 TI - Modulation by phosphorylation of interaction between calmodulin and histones. PMID- 6273220 TI - Coupling between fructose 1, 6-bisphosphatase and myo-inositol synthase: an hypothesis for 'rescue synthesis' of myo-inositol. PMID- 6273221 TI - Purification and identification of calmodulin from Neurospora crassa. PMID- 6273222 TI - [Methyl-3H]thymidine in DNA induces lesions which are recognized by a mammalian DNA-repair endonuclease. PMID- 6273223 TI - Temperature-jump measurement of the spin state relaxation rate of cytochrome P450cam. PMID- 6273224 TI - Biosynthesis of cyclic AMP antagonist in hepatocytes from rats after adrenalin- or insulin-stimulation. Isolation, purification and prostaglandin E-requirement for its synthesis. PMID- 6273225 TI - Dehydro-enkephalins: receptor binding activity of unsaturated analogs of Leu5 enkephalin. PMID- 6273226 TI - The effect of an insulin-sensitive chemical mediator from rat adipocytes low Km and high Km cyclic AMP phosphodiesterase. PMID- 6273227 TI - EPR spectral stimulation on cluster N-1b in NADH-ubiquinone oxidoreductase of bovine heart mitochondria. PMID- 6273228 TI - An electrochemical interpretation of metabolism. PMID- 6273229 TI - Subcellular distribution of the subunits of cyclic AMP-dependent protein kinase isoenzymes in human tonsillar lymphocytes. PMID- 6273230 TI - A new sequence-specific endonuclease from Streptococcus durans. PMID- 6273231 TI - Effect of the Na+ ionophore monensin on basal and noradrenaline stimulated gluconeogenesis in rat renal tubule fragments. PMID- 6273232 TI - Binding of 1,25-dihydroxy-[3H]vitamin D3 in nuclear and cytosol fractions of whole mouse skin in vivo and in vitro. PMID- 6273233 TI - Ionic and metabolic bases of neuronal thermosensitivity. PMID- 6273234 TI - Neurophysiology of fever. AB - Fever is a primary disorder of thermoregulation and a common clinical sign in many diseases. It is characterized by an upward displacement in the level at which body temperature is regulated. Early attempts to study hypothalamic neuronal activity in relation to fever described the behavior of isolated single units after intravenous injections of endotoxin pyrogen. It was concluded that the thermosensitivity of many warm-sensitive units was depressed after pyrogen injections, but due to the indirect technique employed, it is not possible to distinguish whether this observation is the cause or result of fever. A decrease in hypothalamic thermosensitivity is contrary to observations made during fever in conscious animals. More specific applications of pyrogenic stimuli such as prostaglandin E1 onto individual hypothalamic neurons using the technique of microelectrophoresis have not borne out these earlier observations. A major obstacle to studying the neurophysiology of thermoregulation and fever is the absence of any obvious correlation between neuroanatomy and function in the hypothalamus. Present methods of identifying and classifying hypothalamic cells as participants in thermoregulation are patently inadequate. Until a more specific correlation between anatomy and function is established, the neurophysiological mechanisms of fever will remain obscure. PMID- 6273235 TI - Hypothalamic mechanisms in thermoregulation. AB - Certain preoptic and rostral hypothalamic neurons are sensitive to changes in local preoptic temperature (Tpo). These neurons also receive much afferent input from peripheral thermoreceptors and control a variety of thermoregulatory responses. In thermode-implanted animals, preoptic warming increases the firing rate in warm-sensitive neurons and elicits heat loss responses such as panting and sweating. Preoptic cooling increases the firing rate in cold-sensitive neurons and elicits, first, heat retention responses (e.g., cutaneous vasoconstriction and thermoregulatory behavior), then heat production responses (e.g., shivering and nonshivering thermogenesis). It is likely that the preoptic thermosensitive neurons control these thermoregulatory responses because both respond similarly to changes in Tpo and skin temperature. Specifically, skin warming not only increases panting, skin blood flow, and the firing rate of warm sensitive neurons, but also decreases the sensitivity of all these responses to Tpo changes. Skin cooling not only increases metabolic heat production, heat retention behavior, and the firing rate of cold-sensitive neurons, but also increases the hypothalamic thermosensitivity of all these responses. Low-firing warm-sensitive neurons receive little afferent input and are most sensitive to high Tpo. Many of these low-firing neurons probably serve in controlling heat loss responses. High-firing warm-sensitive neurons receive much excitatory afferent input and are usually sensitive only to low Tpo. These neurons probably exert their greatest influence on heat production responses, possibly by inhibiting and, thus, determining the thermosensitive characteristics of nearby cold-sensitive neurons. PMID- 6273236 TI - Calcium antagonism is no rose. AB - Much remains to be defined about the mechanism of action of calcium entry blockers. The diversity of their pharmacological actions reflects the many effects of the calcium ions which they block. The observation that D-600 decreases myogenic tone in the rabbit basilar artery, whereas it increases this tone in the facial vein, is cited as an example of this diversity. PMID- 6273237 TI - Calcium entry blockers and myocardial function. AB - Ca2+ enters myocardial cells through a variety of pathways, including in exchange for Na+; by passive diffusion; through voltage-activated, gated channels; and in exchange for K+, Ca2+ entry through the voltage-activated channels is an essential step in excitation-contraction coupling. It is only this component of Ca2+ transport that is inhibited by the Ca2+ entry blockers. As a group, therefore, these drugs interfere with excitation-contraction coupling in heart but not in skeletal muscle. Accordingly they reduce the energy requirements of the heart. Their inhibitory effect on voltage-activated inward transport of Ca2+ into smooth muscle cells also results in dilation of the coronary vessels, with improvement in coronary perfusion, and of peripheral vessels, with after-load reduction. The resultant action of these drugs in maintaining myocardial energy balance and intracellular Ca2+ homeostasis is therefore complex, and tends toward preservation of myocardial structure and function after episodes of ischemia. Although the Ca2+ entry blockers prevent protein release and preserve ultrastructure in damaged myocardium, this is probably an indirect effect of their ability to impede slow channel transport of Ca2+. PMID- 6273238 TI - Effects of drugs on calcium-related phenomena in red blood cells. AB - Red blood cells (RBCs) keep their internal Ca concentration at a low level by virtue of their limited Ca permeability and a potent, ATP-dependent system for Ca extrusion. Cytoplasmic Ca concentrations can be raised by in vitro procedures such as ATP depletion or the use of certain ionophores. High-Ca cells prepared by these means demonstrate a number of interesting morphological, physical, and biochemical changes. Some of these phenomena can be influenced by drugs, usually in suprapharmacological concentrations. The potential importance of these observations for the treatment of hemolytic or microcirculatory disorders is uncertain, inasmuch as the role of Ca in modifying the properties of RBCs in vivo is not clear. PMID- 6273239 TI - Testosterone and gossypol effects on human sperm motility. AB - Testosterone concentration in seminal fluid has been found to be high in infertile males (75 +/- 11 pg/100 microliter). Fertile males have a testosterone concentration of 29 +/- 3 pg/100 microliter. The effects of adding 50, 150, and 300 pg of testosterone to 100 microliter of ejaculate have been studied by turbidimetric analysis (Sokoloski J, et al. Fertil Steril 28:1337, 1977). This method permits objective measurements of sperm velocity and percentage of rapidly moving sperm in a sample (%RM). A dose-dependent effect of testosterone on sperm motility was seen. Fifty pg/100 microliter had no effect on velocity or percentage of moving sperm; 150 pg of testosterone produced a decrease of 36% +/- 8; and 300 pg/100 microliter, a decrease of 62% +/- 8. Caffeine had a stimulatory effect on the percentage of motile sperm at doses of 400 microliter of semen. Likewise, dibutyryl cAMP (10 microgram/microliter) had a positive effect on sperm velocity. The stimulatory effect of these two drugs were negated when 300 pg of testosterone was added to the preparations. Other steroids (17 alpha testosterone, 17 alpha-estradiol, and 17 beta-estradiol, DHT, and progesterone) tested under the same experimental conditions had no effect on sperm motility, but cottonseed oil (goosypol) had drastic effects. Doses a little as 100 pg/100 microliter produced a 90% decrease in sperm motility. PMID- 6273240 TI - Collagenolytic enzyme activity in human ovary: an ovulatory enzyme system. AB - Collagenolytic enzyme activities presumed to play an important role in ovulation were investigated in the human follicular apex, base, and granulosa cell layer throughout the ovarian cycle. Those analyzed were human ovarian collagenase, 2,4 dinitrophenyl-Pro-Gln-Gly-Ile-Ala-Gly-Gln-D-Arg-OH peptidase, N-carbobenzoxy-Gly Pro-Gly-Gly-Pro-Ala peptidase, and alpha-N-benzoyl-DL-arginine beta-naphthylamide hydrolase. Collagenase activity was also measured in human granulosa cell cultures. The activities of all four enzymes showed a marked preovulatory decrease in the apex. The activities in the apex were slightly higher than those in the base throughout the ovarian cycle. However, the activities in the granulosa cell layer and collagenase activity in the granulosa cell cultures increased toward preovulation and decreased after ovulation. These findings suggest that collagenase is synthesized in the granulosa cells maximally at preovulation and is consumed in the follicular apex at the same time, resulting in collagen degradation and disruption of the follicular wall in collaboration with other collagenolytic enzymes investigated here. PMID- 6273241 TI - Quantitative analysis of the effects of caffeine on sperm motility and cyclic adenosine 3',5'-monophosphate (AMP) phosphodiesterase. AB - Caffeine in high concentrations (2 to 6 mM) has been shown to increase sperm motility and metabolism when added to semen. High concentrations of caffeine have also been demonstrated to inhibit cyclic adenosine 3',5'-monophosphate (AMP) phosphodiesterase isolated from sperm. Since cyclic AMP is known to increase sperm motility, some investigators have attributed the stimulatory effect of caffeine on sperm motility to the inhibition of phosphodiesterase. The authors used semen from healthy donors to quantitate the effects of caffeine on sperm motility by turbidimetric analysis and to analyze kinetically the effect of caffeine on phosphodiesterase isolated from sperm preparations. Caffeine (0.012 to 2.0 mM) produced a dose-dependent increase in sperm motility. The ED50 for motility stimulation was 300 microM of caffeine; a significant increase in motility was observed with 60 microM; and maximal stimulation (a threefold increase) occurred with 1 mM. The phosphodiesterase activity of sperm homogenates displayed linear kinetics with a Vmax of 0.5 nmoles/mg of protein per minute and a Km of 125 microM. Caffeine (0.1 to 40 mM) produced a dose-dependent inhibition of sperm phosphodiesterase. Kinetically, caffeine was a mixed inhibitor of sperm phosphodiesterase, displaying a Ki of 1.2 mM. These results suggest that caffeine may stimulate sperm motility by a mechanism other than phosphodiesterase inhibition. PMID- 6273242 TI - Sympathetic control of brown adipose tissue in the regulation of body weight. PMID- 6273243 TI - Techniques for the microinjection of single cells. PMID- 6273244 TI - In vitro histogenic capacities of limb mesenchyme from various stage mouse embryos. PMID- 6273245 TI - Calmodulin in starfish oocytes. I. Calmodulin antagonists inhibit meiosis reinitiation. PMID- 6273246 TI - Hormones and factors that stimulate growth of a rat islet tumor cell line in serum-free medium. AB - A number of hormones and factors were found to stimulate growth of cultured rat islet tumor cells, the RIN-r cell line. A serum-free supplemented medium from RIN r cells was formulated. It consisted of a 1:1 mixture of Ham's F12 and DME media with the addition of insulin, transferrin, triiodothyronine, prolactin, growth hormone, and an extract of proteose peptone (medium IM). The growth rate of RIN-r cells in this medium is as great as it is in 10% serum-supplemented medium. Up to 10 populations doublings occurred over a period of 20 days. Insulin is a very effective mitogen for RIN-r cells and has an effect on concentrations as low as 30 ng/ml. In addition, the insulin-like somatomedin, multiplication stimulating activity (MSA), is a growth factor at 50 ng/ml. It was found that RIN-r cells proliferate and continue to produce immunoreactive insulin in a hormonally and nutritionally defined medium. This medium is derived from medium IM, in which insulin is replaced with MSA and proteose peptone is omitted. Variations of this medium may prove useful in studies on the growth and function of the normal islets in long-term primary culture. PMID- 6273247 TI - Appearance of LH-immunoreactive cells in the Rathke's pouch of the chicken embryo. AB - The differentiation of the pituitary of the chicken embryo was studied by means of an immunohistochemical technique using antisera to turkey and chicken pituitary hormones. Immunoreactive LH-cells are detected in 4-day embryos (stage 23 of Hamburger and Hamilton) when the primordium of the anterior pituitary, the Rathke's pouch is only composed of a single-layer epithelium lined with an undifferentiated mesenchyme. A few immunoreactive cells are observed grouped on the posterior aspect of the pouch. As development proceeds, a strip of positive cells is detected encircling the Rathke's pouch. Prolactin-, growth hormone-, and ACTH-immunoreactive cells are detected in 6- and 7-day embryos, only after the pituitary has acquired its characteristic structure with cords in which different cell types become progressively recognizable. The early appearance of immunoreactive LH-cells following a precise distribution shows that secretory properties and differentiation capacities are acquired simultaneously in the epithelium of the Rathke's pouch and may be induced by the same stimulus. PMID- 6273248 TI - Curing of virus persistent infection in HVJ (sendai virus) carrier hamster tumor cells by transplantation. AB - Transplantability of hamster tumor cells in the syngeneic system was considerably reduced by HVJ (Sendai virus) persistent infection of the cells, but these HVJ carrier cells still showed tumorigenicity, especially at high transplantation doses. Therefore, the existence of HVJ antigens or infectious HVJ in the tumors formed in vivo by transplanted HVJpi (temperature-sensitive HVJ) or HVJo (non temperature-sensitive HVJ) carrier tumor cells was examined in relation to the reduced transplantability of the cells. In stamp preparations of tumors cells, no HVJ antigens were detected from 14 days after transplantation, though they were positive if observed within 7 days. Infections HVJ particles could also be recovered from tumor tissues which were cocultivated with LLCMK2 cells within 7 days after transplantation. However, tumor cells recultured in vitro after more than 3 weeks post transplantation did not show HVJ antigens, even after various attempts at HVJ induction such as culture at low temperature, serial passages in vitro, treatment with 5-iododeoxyuridine, etc. The cytopathic effects and fluorescent antibody staining of these cells became positive when they were superinfected with wild-type HVJ, indicating a new establishment of HVJ infection after the curing of the HVJ genome initially carried. In contrast, the original HVJ carrier tumor cells were demonstrated to be completely resistant to this superinfection (one of the characteristics of persistent infection). These results seem to show that HVJ persistence in its carrier tumor cells was ultimately cured by transplantation and may provide at least a partial explanation of why the xenogenization of tumor cell by HVJ persistent infection was relatively weak. PMID- 6273249 TI - Metabolism of pyrimidine nucleotides in various tissues and tumor cells from rodents. AB - The metabolism of pyrimidine nucleotides in various tissues and tumor cells of rodents was investigated. Ribonucleoside diphosphate reductase, thymidine monophosphate kinase and DNA polymerase (alpha, beta) were specifically localized in tumor cells, i.e., the activities of these enzymes in tumor cells were at least three times higher than those in normal tissues, including rapidly growing tissues, such as bone marrow, thymus, and spleen. The activities of deoxycytidine monophosphate deaminase and all the nucleoside kinase were high not only in tumor cells, but also in rapidly growing normal tissues, so that these enzymes are unsuitable as targets for cancer chemotherapy. The tissue distribution of other enzymes, including orotate phosphoribosyltransferases, cytidine triphosphate synthetase, thymidine monophosphate synthase, nucleoside phosphorylases and cytidine deaminase had no relation with the cell growth rate. AH130 tumor cells and the thymus showed specific increases in the activities of enzymes involved in de novo DNA synthesis. In contrast, Yoshida sarcoma and bone marrow showed high activities of enzymes in the salvage pathway of DNA synthesis, which suggested that the two tumors have different patterns of nucleotide metabolism. PMID- 6273250 TI - Ultrasound and computed tomographic demonstration of portal vein thrombosis in hepatocellular carcinoma. AB - Two cases of multinodular hepatocellular carcinoma (HCC) in which ultrasound and computed tomography (CT) revealed portal vein thrombosis are presented. The diagnostic value of determining the presence of portal vein thrombosis in patients with suspected HCC is discussed. PMID- 6273251 TI - Production of angiotensin I converting enzyme by liver granuloma macrophages of Schistosoma mansoni infected mice. AB - Angiotensin I converting enzyme activity is detectable in serum and isolated liver granulomas of mice infected with Schistosoma mansoni. At the chronic phase of the infection (20 wk) the intensity of the granulomatous response spontaneously diminishes. Concomitantly, an increase in angiotensin I converting enzyme activity is observed. The objective of this investigation was to determine the source of this elevated angiotensin I converting enzyme activity. In chronically infected mice, measurements showed that angiotensin I converting enzyme activity in hepatic venous blood was higher than that of peripheral venous blood. In contrast, normal mice demonstrated no difference in regional venous angiotensin I converting enzyme activity. Isolated liver granulomas cultured in vitro released angiotensin I converting enzyme into the culture medium. When granulomas were dispersed into single cell suspension, angiotensin I converting enzyme activity was traced to the adherent cell fraction. Both granulomas and adherent cells from granulomas of the chronically (20 wk) infected mice secreted far more angiotensin I converting enzyme than those from animals with acute (8 wk) infection. Cycloheximide partially inhibited enzyme release. We conclude that in murine schistosomiasis, angiotensin I converting enzyme is produced by the granuloma macrophage. The enzyme is released from the granuloma into the circulation probably resulting in increased serum angiotensin I converting enzyme activity. The role of elevated angiotensin I converting enzyme activity within the lesion as well as the circulation remains to be elucidated. PMID- 6273252 TI - Herpes simplex-hepatitus use of immunoperoxidase to demonstrate the viral antigen in hepatocytes. AB - In this paper we describe 2 patients with herpes simplex hepatitis. Submassive liver necrosis occurred in both patients, one of whom survived. Notable in the clinical course of both patients was the almost simultaneous occurrence of three events-fever, marked elevation of serum transaminases, and leukopenia. Using an immunoperoxidase staining technique we demonstrated herpes simplex viral antigen in the nucleus, cytoplasm, and cell membrane of affected hepatocytes. PMID- 6273253 TI - Ischemic injury in the cat small intestine: role of superoxide radicals. AB - Hemorrhagic lesions in the small intestinal mucosa have been demonstrated in humans and experimental animals following hemorrhagic shock and intestinal ischemia. In order to define the role of superoxide radicals and xanthine oxidase in the pathogenesis of the mucosal lesions, we compared the microscopic mucosal changes produced by 3 h of regional hypotension (intestinal arterial pressure = 30 mmHg) in untreated cats and cats pretreated with either superoxide dismutase or allopurinol. In the untreated animals the mucosa was characterized by massive epithelial lifting down the sides of the villi, completely denuded villi, and most frequently by disintegration of the lamina propria, hemorrhage, and ulceration. Pretreatment with either superoxide dismutase or allopurinol significantly attenuated the necrosis of villus and crypt epithelium produced by 3 h of ischemia. The results of this study suggest that superoxide radicals are involved in the pathogenesis of ischemic mucosal lesions and that the enzyme xanthine oxidase is the source of superoxide radicals in the ischemic small bowel. PMID- 6273254 TI - Granular cell tumor of the esophagus. PMID- 6273255 TI - Regulation of catecholamine release by presynaptic receptor system. PMID- 6273257 TI - Inhibition of glycine and GABA receptor binding by several opiate agonists and antagonists. PMID- 6273256 TI - Aliphatic alcohols and the inhibition by thiol chelating agents of the binding of [3H[ dihydroalprenolol to myocardial beta-adrenoceptors. PMID- 6273258 TI - [Artificial Tn2551 transposon with replicative properties]. AB - A hybrid plasmid, pBE10 was constructed. It consists of DNAs of RSF2124 (ColE1 :: Tn3) plasmid and pUB110 plasmid of Staphylococcus aureus. The latter can be stably maintained in Bacillus subtilis. BamHI cleaved pUB110 was introduced into the BamHI site of transposon Tn3 and the resulting enlarged Tn3 (Tn2551) was transposed from pBE10 onto phage lambda and than to pMB9 (Tc) and RSF1010(Sm Su) plasmids. Restriction and heteroduplex analysis of pMB9 :: Tn2551(pBE21) and RSF1010 :: :: TN2551(pBE32) was carried out. Plasmids pBE10, pBE21 and pBE32 demonstrated some kind of molecular instability when introduced by transformation into Bacillus subtilis. PMID- 6273259 TI - Age-related activity changes in actomyosin ATPase and arginine phosphokinase in male Drosophila melanogaster Meig. AB - The specific activity of each of the two enzymes primarily concerned with the energy metabolism of flight muscle contraction was determined in male Drosophila melanogaster, from emergence through 5 weeks postemergence. Ca++-activated actomyosin ATPase activity rose rapidly from a minimum at emergence to a peak level at 4-5 days, falling gradually thereafter to a minimum level at 10-11 days postemergence, with no change thereafter. Arginine phosphokinase activity rose more gradually to a maximum at 11-12 days postemergence and then fell slowly to a minimum at 18-19 days. These data represent a sequential pattern of biochemical changes similar to those previously observed in other dipteran species, confirming the primary role of a genetic program for maturation and senescence of flight ability in holometabolous adult insects at the functional and underlying cellular, biochemical levels. PMID- 6273260 TI - Human papillomavirus and cancer of the uterine cervix. PMID- 6273261 TI - Papillomavirus infections and human genital cancer. PMID- 6273262 TI - Association of human papillomavirus with neoplastic processes in the genital tract of four women with impaired immunity. PMID- 6273263 TI - Herpesvirus antigens as markers for cervical cancer. PMID- 6273264 TI - Genital cancer and viruses. PMID- 6273265 TI - New concepts: vaccines. PMID- 6273266 TI - "The Herpesvirus hypothesis"--are Koch's postulates satisfied? PMID- 6273268 TI - Anti-HLA antibodies detected in the serum of patients with gestational trophoblastic tumors (choriocarcinomas and invasive moles). PMID- 6273267 TI - Experimental studies of carcinogenesis of the uterine cervix in mice. PMID- 6273269 TI - [Morphologic study on cAMP and cGMP distribution in peripheral and bone marrow cells in normal and leukemic subjects. (Peroxidase-antiperoxidase method-PAP) (author's transl)]. PMID- 6273270 TI - Studies on the mechanism of carcinogenicity of diethylstilboestrol: role of metabolic activation. PMID- 6273271 TI - Structure-mutagenicity analysis with the CHO/HGPRT system. PMID- 6273272 TI - Failure of chloroform to induce chromosome damage or sister-chromatid exchanges in cultured human lymphocytes and failure to induce reversion in Escherichia coli. PMID- 6273273 TI - [Current surgical therapy of rectal cancer]. PMID- 6273274 TI - [Studies on the nature of tubular structures as a marker of herpes simplex virus type 2 (author's transl)]. PMID- 6273275 TI - Insulin and glucagon binding to hepatocytes in relation to circulating hormones and metabolites in goats maintained on different diets. AB - Insulin and glucagon receptors were measured on hepatocytes in relation to circulating hormones (insulin, glucagon and growth hormone) and metabolites (non esterified fatty acids, volatile fatty acids, glucose, total lipids, urea and alpha-amino nitrogen), in twelve, two-year old, castrated male goats, fed rations of different composition and dietary energy. The goats were separated into four groups; group 1 was fed a restricted ration of 600 g clover hay/day, group 2 a ration high in carbohydrate (rolled barley), group 3 a ration high in fat (protected tallow) and group 4 a ration high in protein (fish meal). Rations in groups 2 - 4 were fed at 1300 g/day supplemented with 600 g of clover hay. The binding of insulin to hepatocyte receptors was increased by restricting dietary intake when compared to the high energy intake groups (p less than 0.01). There was no significant difference between the insulin binding of groups 2 -4. Glucagon receptor binding was increased on the high protein diet in comparison with th ration high in carbohydrate (p less than 0.05) or in fat (p less than 0.01). The glucagon binding was reduced by restricting feed intake when compared wih feeding high protein (p less than 0.02), but slightly increased when compared with feeding diets high in both carbohydrate or fat (p less than 0.02). There was no significant difference between the high carbohydrate or high fat fed groups. These changes in hormone receptors were accompanied by inverse changes in plasma insulin and glucagon. PMID- 6273276 TI - An early effect of cortisol, previous to its glycogenogenic action. AB - Cortisol produces a glycogenogenic effect 5 hours after intraperitoneal injection to 3 day old chicks. This effect is dependent on protein synthesis because it can be blocked by antibiotics such as actinomycin D. On the other hand, there is a previous glycogenolytic effect 45 minutes after cortisol administration which is independent of protein synthesis. Thyroid hormones produce a similar early effect as has been previously shown. However, the observed glycogenolysis after cortisol injection is not correlated with an enhancement in the liver cAMP levels. PMID- 6273277 TI - Heterogeneous forms of thyroid-stimulating hormone in mouse thyrotropic tumor and serum: differences in receptor-binding and adenylate cyclase-stimulating activity. AB - Thyroid-stimulating hormone (TSH) in tumor and serum of mice with thyrotropic tumors was studied by radioimmunoassay (RIA), radioreceptor assay (RRA) and thyroid adenylate cyclase assay (ACA). In unfractionated samples, serum TSH displayed significantly higher ACA/RIA (0.69 +/- 0.10) and ACA/RRA ratios (0.79 +/- 0.04) than TSH in tumor extracts (0.45 +/- 0.05 and 0.31 +/- 0.01, respectively). After gel chromatography, both tumor and serum TSH activity measured in RIA and RRA eluted in broad, heterogeneous peaks with an apparent molecular weight range of 26,000 - 44,000 daltons. The ACA activity of tumor TSH eluted in 2 sharp peaks (26,000 and 44,000) while that of serum TSH eluted in 3 peaks (26,000, 33,000 and 44,000). The ACA/RIA and ACA/RRA ratios varied greatly among the chromatography fractions, the lowest ratios being detected in the tumor TSH of 33,000 daltons (0.02 for each) and the highest ratios in both tumor and serum TSH of 26,000 (ACA/RIA = 1.60 - 1.90, ACA/RRA = 1.12 - 1.20). Since previous biosynthetic studies have suggested that such heterogeneous forms of mouse tumor TSH differ solely in carbohydrate content, our data suggest that the biologic activity of TSH may be modulated by its glycosylation. PMID- 6273278 TI - Comparison of human growth hormone binding to rat liver plasma and Golgi membranes. AB - The binding characteristics of hGH to Golgi and liver plasma membranes isolated from normal adult female rats have been compared to assess the biological differences between the Golgi and plasma membrane receptor. The effect of cations and the time course of binding were qualitatively similar for both Golgi and plasma membranes. The Golgi membranes from normal rats exhibited maximum binding at pH 6-7 compared to 5-6 for other membrane fractions. The highest apparent affinities (approx. 1 x 10(9) M-1) for hGH were observed in the light Golgi membranes from normal rats and the light and intermediate Golgi membranes from ethanol treated rats. The lowest apparent affinities (0.026 -0.1 x 10(9)M-1 if determined by competitive binding curves or 0.07 - 0.32 x 10(9)M-1 by Scatchard plots) for hGH were observed in plasma membranes isolated by either Neville's or Ray's method or a combination of both methods. The hGH receptors were determined to be lactogenic by competitive binding curves. The affinities of Golgi membranes were not affected by alterations in isolation procedures. Ethanol treatment of the rats prior to sacrifice and membrane isolation resulted in linear Scatchard plots for hGH binding to Golgi membranes compared to curved Scatchard plots for the Golgi membranes of normal rats. The marker enzyme activities of glucose-6 phosphatase and adenylate cyclase were lower in Golgi from ethanol treated rats while the galactosyl transferase activity increased in lighter golgi fractions from ethanol treated rats. PMID- 6273279 TI - Cation transport in intact erythrocytes of hyperthyroid patients: role of the NaK ATPase pump. AB - Studies of erythrocyte (RBC) cation fluxes and concentrations in hyperthyroid subjects have recently been reported with the suggestion that Na-K ATPase activity was decreased. We have studied tha kinetics of total and ouabain sensitive K+ uptake utilizing 86Rb as a tracer in the intact erythrocytes of 7 hyperthyroid subjects and compared the results of those of a healthy control population. We find total K+ transport is depressed in the RBC of hyperthyroid subjects. The Vmax for K+ transport for hyperthyroid subjects is 1.8 +/- 0.17 x 10(-4) mM K+/10(9) RBC/hour versus a control of 2.3 +/- 0.14 x 10(-4) mM K+/10(9) RBC/hour. This depression in Vmax is evident in spite of no significant differences in the Km for the system when hyperthyroid subjects (2.7 +/- 0.19 mM) are compared to controls (2.38 +/- 0.21 mM). Further, the depressed K+ transport appears to be the result of depressed ouabain--insensitive K+ transport. Although the percent of the ouabain-sensitive K+ transport is greater in the hyperthyroid subject (82.5%) versus controls (72.5%), this simply reflects a relative change in a system where total transport is dropping but the ouabain-sensitive component is remaining unchanged. None of these findings can be directly or indirectly related to thyroid hormone and it is suggested that the ion transport changes reflect factors independent of thyroid hormone. PMID- 6273280 TI - Effects of cyclic adenosine 3':5'-monophosphate and cyclic guanosine 3':5' monophosphate on liver proteoglycan synthesis. PMID- 6273281 TI - Mallory body-like abnormalities in carcinomas induced by cultured transformed rat liver cells. AB - Hepatoma cells isolated from rats after administration of a carcinogen, diethylnitrosamine, and propagated in culture, contained a genetically stable cytoskeletal abnormality resembling Mallory bodies. These juxtanuclear aggregates of intermediate-sized filaments were maintained in carcinomas produced in nude mice after inoculation of uncloned mass cultures and a cloned subculture. Paraffin and frozen sections of these tumors revealed acentric nuclei and a glassy hyalin-type cytoplasmic lesion which stained pink with hematoxylin-eosin and blue with Mallory's aniline blue stain. The cells in culture and in the tumor sections were strongly positive for gamma-glutamyl transpeptidase. Cryostat sections examined by indirect immunofluorescence microscopy with antisera to purified bovine hoof prekeratin, desmosome-associated tonofilaments from bovine muzzle, and murine vimentin, as well as transmission electron microscopy revealed the presence of juxtanuclear aggregates of intermediate-sized filaments. All characteristics previously reported for the tissue culture cell line were stably maintained in the tumor tissue. These results suggest that the Mallory body containing cells frequently observed in man in alcoholic hepatitis and other degenerative liver diseases could, under appropriate environmental "promoting" conditions, be precursor cells in focal hepatocellular carcinoma formation. PMID- 6273282 TI - Membrane traffic at the hepatocyte's sinusoidal and canalicular surface domains. PMID- 6273283 TI - Advances in hepatobiliary ultrasonography. PMID- 6273284 TI - [1H-NMR spectroscopy--a potent method for the determination of substrate specificity of sialidases (author's transl)]. AB - We describe here the application of 1H-NMR spectroscopy to determine the substrate specificity of sialidases using a 1:1 mixture of NeuAc alpha 2-3Gal beta 1-4Glc and NeuAc alpha 2-6Gal beta 1-4Glc, one viral and five bacterial sialidases. This method utilizes the separate signals in NMR spectra, characteristic for the different alpha ketosidically linked NeuAc residues and also for bound and free NeuAc. The signals generally most suitable for these purposes are those of H3a, H3e and NCOCH3. By observation and integration of these signals we can follow--qualitatively and quantitatively--which and how many NeuAc residues of the substrates are hydrolized. In contrast to the generally used colorimetric tests it is now possible to investigate with this method substrates containing two or more NeuAc residues and to determine the corresponding rate constants for hydrolysis of the differently bound NeuAc molecules. The six sialidases used show large differences in their specificity as compared with our "model substrate": The sialidase from fowl plague virus hydrolizes NeuAc alpha 2-3Gal beta 1-4Glc nearly 18 times and the enzyme from Clostridium perfringens four times, from Vibrio cholerae two times faster than NeuAc alpha 2-6Gal beta 1-4Glc. On the contrary, the sialidase from Arthrobacter ureafaciens hydrolizes the alpha 2-6 linkage six times faster than the alpha 2-3 linkage. The sialidases from Bifidobacterium show no obvious differences in their specificities relative to the linkage. PMID- 6273285 TI - The metal ion requirement for activation of latent collagenase from human polymorphonuclear leucocytes. AB - Latent human PMN leucocyte collagenase (enzyme-inhibitor complex) was shown to require zinc for the property of being activatable by various disulfides [see Macartney, H.W. and Tschesche, H. (1980) FEBS Lett. 119, 327--332]. The active enzyme also requires zinc for activity, indicating a possible participation in the enzyme's reaction mechanism and/or stabilization of the active site. The zinc in the latent enzyme may be removed by dialysis against EDTA, or cysteine. This produces a zinc-free latent enzyme which cannot be activated by any of the disulfide-containing activators. Readdition of zinc to the EDTA-inhibited latent enzyme, at the same concentration as the EDTA, produces an activatable latent enzyme once again. However, excessive zinc concentrations (more than three times the concentration of EDTA) exhibited an inhibitory effect on the activation process. Thereafter the inhibitor cannot be removed by disulfides from the enzyme inhibitor complex of the latent enzyme. The zinc in the latent enzyme may be replaced by other double-positive metal ions such as cobalt, manganese, magnesium and copper. PMID- 6273286 TI - The effect of trifluoroacetyl-cytochrome c on the cytochrome c/cytochrome c oxidase reaction. AB - The importance of electrostatic interactions to the reaction between cytochrome c and cytochrome c oxidase is indicated most directly by the rapid increase in Km as ionic strength is increased. However, Chessa et al. (1980, Hoppe-Seyler's Z. Physiol. Chem. 361, 1077--1091) have recently found that a cytochrome c derivative trifluoroacetylated at all 19 lysine amino groups decreased the reaction rate between ferrocytochrome c and cytochrome c oxidase nearly as much as native ferricytochrome c, a known inhibitor that binds cytochrome c oxidase. They suggested that this was due to the formation of an inhibitory complex between trifluoroacetyl-cytochrome c and cytochrome c oxidase, which would argue against the importance of electrostatic interactions to the native complex. We have demonstrated that this apparent inhibition is in fact caused by a rapid electron transfer between native ferrocytochrome c and trifluoroacetyl ferricytochrome c which decreases the concentration of native ferrocytochrome c, thus decreasing the reaction rate. PMID- 6273287 TI - [Steric requirements for the carboxymethylation of His 12 in ribonuclease A studied by 2'- and 3'-bromoacetylesters of modified nucleosides (author's transl)]. AB - The 2'- and 3'-bromoacetylesters of different nucleosides have been synthesized and tested in the carboxymethylation of His 12 of pancreatic ribonuclease A. The 2'-esters of ribosides react more than 100 times faster compared with 3'-esters or 2'-arabinosides. 2'(3')-esters of nucleosides which are not substrates (adenosine, N3-methyluridine or 6-methyluridine) do not react, obviously due to an inexact orientation of the base. The difference spectrum obtained after the reaction of 2'-acetamido-2'-deoxyuridine with ribonuclease A is identical with the difference spectrum of 2'-cytidylic acid, thus indicating that the base occupies the same position as in the binding of substrate analogous inhibitors. Immobility of His 12 and its reaction with the CH2-Br group leads to a disagreement with the concept of a base catalysis of His 12 at the 2'-OH-group in the reaction with substrates. The pH dependence of the reaction is identical with the carboxymethylation of His 12 and 119 using bromoacetate. Therefore, binding to His 119 cannot be considered when interpreting the bell-shaped curve, which might be explained by the alternative, the existence of a triprotonated diimidazole system between His 119 and His 12. No activity can be detected for His-12-carboxy-methylated ribonuclease in all pH ranges. PMID- 6273288 TI - Synthesis and preliminary biochemical characterization of spin-labeled derivatives of ATP and its 'non-cleavable' analog, adenosine 5'-beta, gamma methylenetriphosphate. PMID- 6273289 TI - A critique of the ACE test. PMID- 6273290 TI - Clinicopathological studies of liver cirrhosis and hepatocellular carcinoma in a general hospital. AB - Clinicopathological studies of 181 patients with liver cirrhosis undergoing autopsy in the pathology department of a municipal hospital between 1973 and 1976 are reported. The main etiological types were alcoholic (25.4 per cent), HBsAg positive (14.9 per cent), and cryptogenic cirrhosis (54.7 per cent). Four patients had multifactorial and secondary biliary cirrhosis and one patient had congestive cirrhosis. The morphological characteristics of the condition and the age and sex distribution of the patients were analyzed in each etiological group. Hepatocellular carcinoma occurred in 28.7 per cent of the cases, most frequently in association with HBsAg positive cirrhosis. Hepatocellular atypia was significantly more frequent in HBsAg positive than in other etiological types of cirrhosis and significantly more frequent in cases associated with hepatocellular carcinoma than in those not associated with it. PMID- 6273291 TI - Mucin producing islet cell adenoma. AB - A well granulated beta cell adenoma of the pancreas was found incidentally at autopsy in a patient with a ruptured aortic aneurysm. The tumor was studied by histochemical an immunocytochemical methods. It consisted of a trabecular arrangement of insulin storing cells intermingled with small ducts. Both components were tightly contiguous. Goblet cells were also present within the tumor cell nests. The implication of the growth of the different types of cells is discussed in relation to the concept of an APUD system. PMID- 6273292 TI - Maturation of B-cell differentiation ability and T-cell regulatory function in infancy and childhood. PMID- 6273293 TI - An analysis of effector T cell generation and function in mice exposed to influenza A or Sendai viruses. PMID- 6273294 TI - Cell membrane antigens recognized by anti-viral and anti-trinitrophenyl cytotoxic T lymphocytes. PMID- 6273295 TI - Organization and expression of murine immunoglobulin genes. PMID- 6273296 TI - Human antibody genes: V gene variability and CH gene switching strategies. PMID- 6273298 TI - Mouse liver cell culture. I. Hepatocyte isolation. AB - A method for isolation of mouse liver cells by a two-step perfusion with calcium and magnesium-free Hanks' salt solution followed by a medium containing collagenase is described. Several variations of the commonly used procedure for rat liver cell isolation were quantitatively compared with respect to cell yield and viability. The optimal isolation technique involved perfusion through the hepatic portal vein and routinely produced an average of 2.3 x 10(6) viable liver cells/g body weight. Optimal perfusate collagenase concentration was found to be 100 U of enzyme activity per milliliter of perfusate. Light and electron microscopic evaluation of liver morphology after several steps of the isolation showed distinct morphologic changes in hepatocytes and other liver cells during perfusion. After perfusion with Hanks' calcium- and magnesium-free solution, many hepatocytes exhibited early reversible cell injury. These changes included vesiculation and slight swelling of the endoplasmic reticulum as well as mitochondrial matrix condensation. Subsequent to perfusion with collagenase, the majority of hepatocytes appeared connected to one another only by tight junctional complexes at the bile canaliculi. Multiple evaginations were seen on the outer membrane resembling microville and probably represented the remains of cell-to-cell interdigitations between hepatocytes and sinusoidal lining cells from the space of Disse. The cytoplasmic injury seen after Hanks' perfusion was reversed after collagenase perfusion. After mechanical dispersion, isolated mouse hepatocytes were spherical in shape and existed as individual cells; many (80 to 85%) were binucleated under hase contrast light microscopy. By electron microscopy, cells appeared morphologically similar in cytoplasmic constitution to that seen in intact nonaltered liver cells. PMID- 6273299 TI - Solubilization & properties of an adenosine triphosphatase of reo virus. PMID- 6273297 TI - Effect of sodium butyrate on the hepatoma cell cycle: possible use for cell synchronization. AB - Exposure of HTC cells to sodium butyrate caused inhibition of growth. The site of growth inhibition was studied by time-lapse cinematography and [3H]thymidine incorporation studies. Evidence is presented that sodium butyrate affected the cell cycle at a specific point immediately after mitosis. Inasmuch as it does not modify the interphase duration after its removal, butyrate may be used for HTC synchronization. PMID- 6273300 TI - Studies on the bicuculline-GABA interactions on neuronal membrane with fluorescent labelled anti-bicuculline antibodies. PMID- 6273301 TI - Plasma cAMP and cAMP-phosphodiesterase (PDE) levels in cancer patients before and after surgery. PMID- 6273302 TI - Mixed Mullerian tumour of ovary in association with breast carcinoma and serous cystadenocarcinoma carcinoma of other ovary. PMID- 6273303 TI - Adrenaline influence on the immune response. II. Its effects through action on the suppressor T-cells. AB - Experiments were carried out to specify the adrenaline target among the immunocompetent cells. Adrenaline administered for some hours exerted opposite effects on the natural PFC and RFC: the first were enhanced and the second significantly reduced. These paradoxical results were interpreted as a consequence of the inhibition of the suppressor T-cells in the resting status. Adrenaline appeared to act on the sensitive cells through beta-rather than through alpha-receptors. Further experiments on the adrenaline influence on the syngeneic barrier phenomenon and on the cellular balance at its termination seemed to indicate that adrenaline was directly inhibitory for the Ts but not for their precursors. These results are discussed in the light of the cellular networks regulating the immune response. PMID- 6273304 TI - Dermatosis of viral origin in children. PMID- 6273305 TI - Purification & characterization of two forms of 5'AMP nucleotidase form Neurospora crassa. PMID- 6273306 TI - Regulation of envelope-growth in Escherichia coli: horizontal envelope-growth by a process under cyclic AMP control. PMID- 6273307 TI - A note on the attenuation of buffalopox virus. PMID- 6273308 TI - Effect of ambient temperature on the cell-mediated immune response in mice immunized with herpes simplex virus. PMID- 6273309 TI - The reaction of Funambulus tristriatus tristriatus Rattus blanfordi and Suncus murinus to Kyasanur forest disease virus. PMID- 6273310 TI - A serological survey of pigs in India for swine influenza. PMID- 6273311 TI - Indirect fluorescent antibody test in the differential diagnosis of hepatic amoebiasis from non-specific hepatomegaly. PMID- 6273312 TI - Use of glutaraldehyde-treated sheep erythrocytes in indirect haemagglutination test for amoebic coproantibody. PMID- 6273313 TI - Ability of autoclaved BCG and extracts to induce aspermatogenesis. PMID- 6273314 TI - Cell-mediated immunity and lymphocyte populations in experimental Argentine hemorrhagic fever (Junin Virus). AB - Guinea pigs infected with the XJ prototype strain of Junin virus reproduce the main features of Argentine hemorrhagic fever, showing hemorrhages, leukothrombocytopenia, and focal lymphoid tissue necrosis. Viral lymphotropism is shown by the presence of viral antigens, severe cytopathic effect, and high virus titers in lymphoid organs. A pronounced depression of humoral immune response to sheep erythrocytes as well as to the virus is described. This study was carried out to determine whether cellular immune response was also modified and which cell populations were affected. Delayed hypersensitivity skin reaction to purified protein derivative was found to be markedly depressed after infection. A noticeable decrease in both percentages and absolute T lymphocyte numbers, detected by E rosettes, in spleen and lymph nodes, together with a low absolute T cell number in peripheral blood, were observed. Total cell counts in spleen, lymph nodes, and peripheral blood were also reduced. On the contrary, no modification in percentages of B lymphocytes, as measured by EAC rosettes, was found. These results indicate that cell-mediated immunity is markedly impaired in guinea pigs infected with the XJ strain of Junin virus. Its relationship with the pathogenesis of the disease is discussed. PMID- 6273315 TI - Immunization with major outer membrane protein (porin) preparations in experimental murine salmonellosis: effect of lipopolysaccharide. AB - A crude major outer membrane (porin) preparation, obtained from a rough strain of Salmonella typhimurium earlier shown to be protective both in active and passive immunization of mice against challenge with smooth S. typhimurium, was further purified. Removal of the main impurities, lipopolysaccharide (LPS) and lipoprotein, was accompanied by loss of protective capacity in passive immunization experiments. Reconstitution with rough LPS restored the protective capacity. Protection was, however, concluded not to be due to anti-LPS, because a large fraction of the anti-LPS antibodies could be removed from the protective rabbit antiserum with an LPS immunosorbent without loss of protection. PMID- 6273316 TI - Molecular epidemiology of adenoviruses: alternating appearance of two different genome types of adenovirus 7 during epidemic outbreaks in Europe from 1958 to 1980. AB - Four genome types of adenovirus serotype 7 (Ad7, Ad7a, Ad7b, and Ad7c) were identified by use of DNA restriction enzymes BamHI, EcoRI, and SmaI. We obtained information on the distribution of the four Ad7 genome types by typing 68 Ad7 isolates obtained in the Netherlands from 1958 to 1979 and 36 Ad7 isolates obtained in Sweden from 1964 to 1980. The Ad7 prototype was never detected, and only seven isolates were genome-typed Ad7a. Ad7b and Ad7c accounted for 94% of the genome-typed isolates obtained from patients. Ad7c was isolated in the Netherlands during 1958 to 1969, but only Ad7b has been detected there since 1970. In Sweden, Ad7c was recovered between 1964 to 1972, but only Ad7b has been isolated there since 1973. In conclusion, the newly recognized Ad7b and Ad7c genome types, which in the Netherlands and Sweden represent virtually all genome typed isolates from patients, show a mutually exclusive appearance. PMID- 6273317 TI - Isolation of a protein-containing cell surface component from Streptococcus sanguis which affects its adherence to saliva-coated hydroxyapatite. AB - The isolation and partial characterization of a protein-containing cell surface component from Streptococcus sanguis which blocks the adherence of this microbe to saliva-coated hydroxyapatite are described. Several methods of extraction were attempted. Sonication of whole cells and cell walls proved to be the most successful and yielded biologically active adherence-blocking components. The adherence-blocking ability of these components was effective in intraspecies blocking experiments. The extract obtained from cell walls of S. sanguis was examined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and shown to contain one major and two to three minor bands when stained with Coomassie blue. The molecular weight of the major band was estimated to be 70,000 to 90,000. Gel filtration of the sonified cell wall extract on 10% agarose yielded two active adherence-blocking peaks, the void volume and a second peak. PMID- 6273318 TI - Cytomegalovirus-specific cytotoxicity mediated by non-T lymphocytes from peripheral blood of normal volunteers. AB - The cytotoxicity of circulating mononuclear cells from normal volunteers was determined using human leukocyte antigen (HLA)-typed, low-passaged human skin fibroblasts infected with cytomegalovirus as target cells. Peripheral blood lymphocytes from both seropositive and seronegative individuals possessed virus specific cytotoxicity. Although all target cells used were susceptible to virus specific lysis, lymphocytes from some individuals were more active against some target cells than others. This differential cytotoxicity did not follow a consistent pattern of HLA restriction. Some variations in cytotoxic activity were noted on sequential studies of individual volunteers. Studies of fractionated lymphocytes from selected immune and nonimmune individuals demonstrated that cytotoxicity of lymphocytes from both groups was mediated by nonadherent, Fc receptor bearing cells which did not form rosettes with sheep erythrocytes. Repeated washing sometimes decreased cytotoxicity of lymphocytes from immune individuals, and addition of serum containing antibody to cytomegalovirus enhanced cytotoxicity, suggesting antibody dependence. It is concluded that cytotoxic lymphocytes from nonimmune volunteers possessed characteristics of natural killer cells, whereas those from immune volunteers probably consisted of both natural killer cells and antibody-dependent killer cells. PMID- 6273319 TI - Spontaneous and induced herpesvirus genome expression in Marek's disease tumor cell lines. AB - We incubated 31 newly established Marek's disease tumor cell lines at 41 degrees C for 48 h after subculturing and then examined them to determine the spontaneous rates of expression of viral internal antigen(s), viral membrane antigen(s), and virus isolation. All but two of the lines were isolated from tumors induced by clone-purified Marek's disease virus strain JM-10, GA-5, RB-1B, and BC-1A in nine different genetic strains of chickens with defined histocompatibility antigens. The line-to-line variations in the rates of spontaneous expression for the antigens or virus rescue were great, but the levels of expression were very low in most cases. The median rates of expression for viral internal antigen, viral membrane antigen, and virus isolation were 32, 8, and 2 positive cells per 10(5) cells, respectively (ranges, 0 to 20,280, 0 to 22,990, and 0 to 220 positive cells per 10(5) cells, respectively). The ratio of viral internal antigen expression to virus isolation was extremely variable and often high, whereas the ratio of viral internal antigen to viral membrane antigen expression was more consistent and generally low. The virus strain which induced the cell line influenced the level of virus genome expression, but the cell genotype did not. Cell lines transformed by JM-10 virus, which exhibited low oncogenicity, had significantly (p less than 0.01) higher rates of expression than cell lines transformed by CA-5 and RB-1B viruses, which exhibited high oncogenicity. Treatment with iododeoxyuridine or incubation at 37 degrees C induced increased rates of expression in most lines but not in all lines. The degree of enhanced expression was inversely proportional to the rate of spontaneous expression. PMID- 6273320 TI - Involvement of cells of hematopoietic origin in genetically determined resistance of Syrian hamsters to vesicular stomatitis virus. AB - Susceptibility of Syrian hamsters of the inbred LSH and MHA strains to injection of as few as 10 plaque-forming units of vesicular stomatitis virus (VSV) was shown to occur only after intraperitoneal and intrapleural injection and not after injection of VSV intravenously, intranasally, or in the footpads. Despite the fact that fewer LSH hamsters died when VSV was injected via the latter routes, the histopathology of the VSV-induced disease at early times after infection was identical irrespective of the route of virus administration. Histological examination of tissues at various times after administration of VSV by the various routes revealed that VSV exhibited tropism for lymphoreticular tissue, with the greatest amount of necrosis in the splenic periarteriolar lymphoid sheath. A similar pattern also was observed in VSV-infected tissues from genetically resistant UT1 hamsters. Infectivity titrations of various tissues at different times after intraperitoneal injection of VSV revealed that resistant UT1 hamsters began to clear virus from tissues between 40 and 48 h postinfection, whereas virus titers remained high in susceptible animals. Resistance of UT1 hamsters appeared to require an intact spleen since survival of splenectomized animals was less than that of sham-splenectomized UT1 controls. Sublethal whole body irradiation was also able to reduce resistance of UT1 hamsters (survival was reduced from 100 to 50%). Bone marrow cells from resistant (UT1 X LSH) F1 females were transferred into lethally irradiated susceptible LSH hamsters, and hematopoietic chimeras were produced. After intraperitoneal injection of 100 plaque-forming units of VSV, all of the female chimeras survived, but only 33% of male chimeras survived. These data indicate that resistance to VSV in Syrian hamsters is mediated, at least partially, by cells of hematopoietic origin. PMID- 6273321 TI - Resistance of peripheral autonomic neurons to in vivo productive infection by herpes simplex virus mutants deficient in thymidine kinase activity. AB - We used a model involving acute and latent herpes simplex virus (HSV) infections of mouse superior cervical ganglia to assess in vivo neuronal infections with two thymidine kinase-deficient (TK-) mutants of HSV type 1. Despite replication of the TK- HSV strains at the site of inoculation in the eyes, little if any viral replication occurred in the superior cervical ganglia, as assessed by the viral titers of ganglion homogenates, viral antigens in tissue sections, and histopathological evidence of cytopathology or inflammation. Cyclophosphamide induced immunosuppression and treatment with 6-hydroxydopamine, which enhanced productive infections of superior cervical ganglia with TK+ HSV did not induce TK HSV ganglionic infections. Latent TK- HSV infections were not detected by cocultivation of ganglion explants. Efforts to infect ganglia in culture after they were removed from animals indicated that superior cervical ganglia, and particularly their neuronal elements, resisted productive TK- HSV infection. These results supported the hypothesis that viral thymidine kinase facilitates acute and reactivated productive HSV infections of neurons. PMID- 6273322 TI - Effects of lectins on peripheral infections by herpes simplex virus of rat sensory neurons in culture. AB - Concanavalin A and wheat germ agglutinin are capable of preventing a productive peripheral infection of dissociated rat sensory neurons in culture by herpes simplex virus type 1. Concanavalin A binds to the herpes simplex virion, rendering it inactive, whereas wheat germ agglutinin binds to the peripheral neuritic extensions of the sensory neurons, rendering them incapable of initiating a productive viral infection. This latter effect (i) seems to be specific for wheat germ agglutinin since other lectins have no effect, (ii) is not the result of cellular cytotoxicity, (iii) is dependent on an N acetylneuraminic acid moiety, and (iv) may be due either to viral receptor site masking or to binding of wheat germ agglutinin to the neuritic receptor molecule for herpes simplex virus. PMID- 6273323 TI - Adherence and Streptococcus mutans infections: in vitro study with saliva from noninfected and infected preschool children. AB - An in vitro adherence experiment was designed to mimic the transmission of Streptococcus mutans from mother to child to test the hypothesis that differences in initial adherence reflect differences in susceptibility to infection. The data show that the pretreatment of S. mutans cells with the saliva of the mother in a mother-child pair and the pretreatment of spheroidal hydroxyapatite with that of the child may result in combinations which counteract or foster the initial adherence to a varying extent. The findings indicate that such combinations may determine the risk of S. mutants infection. PMID- 6273324 TI - Quantitative electron microscopy of sensitized rat mast cells. PMID- 6273325 TI - Comparison of the eosinophil chemotactic factor (ECF) with endogeneous hydroxyeicosatetraenoic acids of leukocytes. PMID- 6273326 TI - Proton induced reactions on natural Pb targets. A potential new cyclotron method for 201 Tl production. PMID- 6273327 TI - Effects of carcinogen and/or mutagen on normal and gonatotropin-primed ovaries of mice. AB - The influence of pregnant mare serum gonadotropin (PSGM) on the induction of ovarian tumors by carcinogen(DMBA) and/or mutagens (EMS and BMS) has been studied in Swiss albino mice. Priming of the ovaries of 6-week-old mice with PMSG (50 IU/mouse) 24 h before intragastric intubation of DMBA (5 mg/mouse) resulted in a significant increase in ovarian tumors when harvested 6 months after the treatment. A similar experiment done on 8-week-old mice yielded confirmatory results. Mutagens (EMS and BMS) failed, at given dose levels (200 mg/kg and 300 mg/kg body weight), to induce tumors in normal as well as PMSG-primed ovaries. Pretreatment of animals with PMSG-enhanced DMBA-induced oocyte depletion and also increased the DMBA-3H activity in the ovaries. The augmentation of DMBA-induced tumors in PMSG-primed ovaries could be due to increased uptake of DMBA and increased depletion of oocytes. It is also possible that PMSG-primed ovaries provided more proliferating granulosa cells (and macromolecules) for the interaction of DMBA (or its metabolites) and thereby increased the chances of cell transformation in the ovaries. PMID- 6273328 TI - Differentiation antigens expressed by epithelial cells in the lactating breast are also detectable in breast cancers. AB - Two monoclonal antibodies, 3.14.A3 and 1.10.F3, raised against a delipidated preparation of the human milk fat globule and characterized as epithelium specific (Taylor-papadimitriou et al., 1981) were assayed histologically, by an indirect immunoperoxidase technique, against formalin-fixed, paraffin-embedded, normal and tumour tissue sections, in order to establish their in vivo specificity. Both antibodies bound to less than 10% of the alveoli and ducts in the resting breast, but bound to all areas of alveoli, ducts and secretion in the lactating breast. Binding was to the luminal surface of the alveolar and ductal epithelium. Antibody 3.14.A3 showed positive reactions with each of 20 primary breast carcinomas tested, and with metastatic lesions in lymph nodes from six of these. Antibody 1.10.F3 also reacted with most of the primary carcinomas but not with those of the mucoid type nor with metastatic lesions in lymph nodes. When tested against a variety of normal tissues, 1.10.F3 bound only to the luminal epithelial surface of classically defined exocrine glands, to their associated ducts and to the collecting tubules of kidney and bronchioles of the lung. 3.14.A3 showed a similar pattern of binding to 1.10.F3 but also bound to sweat glands, the alveolar epithelium of lung and the luminal epithelium of the ductuli efferentes of the epididymis. The only tumours, other than breast, showing a positive reaction with the antibodies were adenocarcinomas of the lung, uterus and ovary. PMID- 6273329 TI - Primary hepatocellular carcinoma in Alaskan natives, 1969-1979. AB - The records of 20 Alaskan Native patients with primary hepatocellular carcinoma (PHC) diagnosed in the 11-year period 1969-1979 were reviewed. The annual incidence of PHC was found to be high among Alaskan Native males and especially high among Alaskan Eskimo males (7.6 and 11.2 per 100,000 respectively) in comparison to Greenland and Canadian Eskimos and US white males. Familial and geographic clustering of PHC patients was noted in areas known to be hyperendemic for hepatitis B virus (HBV) infection. A bimodal age distribution among PHC patients occurred with peaks at 15-25 years and 40-65 years. A high prevalence of hepatitis B surface antigen (HBsAg) in serum of patients in the younger age group suggests that HBV infection might be a factor associated with the development of PHC in young Eskimos. PHC in Alaskan Natives is apparently not closely associated with alcoholic cirrhosis. PMID- 6273330 TI - Characterization of clones of tumorigenic feline cells transformed by a chemical carcinogen. AB - Tumorigenic clonal lines derived from soft agar colonies induced by DMBA transformed feline embryo cells were isolated and characterized. The morphologically altered clonal cells formed large aggregates, growing in this aggregate form when suspended in liquid growth medium above an agar base and forming colonies in soft agar with high efficiency. When inoculated into athymic nude mice, chemically altered clonal cells produced progressively growing sarcomas. Cells established from the tumors morphologically resembled the DMBA transformed feline embryo cells and were characterized as cat cells by karyological analysis. The tumorigenic lines were negative for feline oncornavirus-associated cell membrane antigen (FOCMA), and for "gag-X" the transformation-related polyprotein which is encoded by the replication defective feline sarcoma virus. PMID- 6273331 TI - EBV-positive Burkitt's lymphoma from Algeria, with a three-way rearrangement involving chromosomes 2, 8 and 9. AB - Burkitt's lymphoma was diagnosed in male Algerian child presenting with abdominal and jaw tumours. Viral studies revealed that this non-endemic case was associated with the Epstein-Barr virus. Cytogenetic examination of tumour cells showed a complex three-way rearrangement involving chromosomes 2, 8 and 9. Our report indicates that, in Burkitt's lymphoma, not only simple but also complex translocation involving chromosome 8 may arise. This finding in a North African case emphasizes the need for more detailed study of the clinical, virological and cytogenetic features of Burkitt's lymphoma throughout the world. PMID- 6273332 TI - Diploid human lymphoblastoid and Burkitt lymphoma cell lines: susceptibility to murine NK cells and heterotransplantation to nude mice. AB - Human lymphoid cell lines which had been classified on the basis of studies on clonality and morphological, on the basis of studies on clonality and morphological, chromosomal and functional parameters as lymphoblastoid cell line (LCL) of presumed non-neoplastic origin and Burkitt lymphoma (BL) lines of proven malignant origin, were tested for susceptibility to natural killer (NK) cells obtained from the spleens of athymic nude mice. The 20 lines included normal diploid LCL and aneuploid BL lines. All cells carried the Epstein-Barr virus (EBV) genome. In addition, two EBV-negative BL lines were tested. The pronase induced release of 14C-DNA from 14C-thymidine-labelled target cells was used to assess the sensitivity of the cell lines to NK activity. When attempts were made to correlate the growth of the EBV-positive LCL and the EBV-positive BL cell lines in the subcutaneous space of adult nude mice with their susceptibility to NK cells, no significant correlation was observed. The EBV-negative BL cell line, Ramos, however, could be transplanted subcutaneously in nude mice and was more resistant to NK activity than was the EBV-negative BL cell line, BJAB, which cannot be transplanted subcutaneously. Growth of heterotransplanted EBV-converted cell lines in the subcutaneous space of adult nude mice may be influenced by immune effectors other than NK cells. PMID- 6273333 TI - Kaposi's sarcoma. IV. Detection of CMV DNA, CMV RNA and CMNA in tumor biopsies. AB - In order to determine whether human cytomegalovirus- (CMV) DNA homologous sequences as well as CMV-specific RNA(s) and antigen(s) exist in tumor biopsies of Kaposi's sarcoma (KS) DNA-DNA reassociation, RNA-DNA in situ cytohybridization and anticomplement immunofluorescence test (ACIF) tests were applied. Three of 10 DNAs extracted from Kaposi sarcoma biopsies contained DNA sequences homologous to radioactively labelled human CMV DNA probe. The amount of CMV DNA in these sarcoma tissues was calculated to range from 0.7 to 1 genome equivalent per diploid cell. The presence of virus-specific RNA was also demonstrated in section from five of 10 tumor biopsies. CMV-determined nuclear antigen(s) CMNA) present in variable degrees were also demonstrated. In contrast, we could not detect any herpes simple virus type II (HSV-2) or Epstein-Barr virus (EBV) DNA sequences in DNA of these tumor biopsies. Furthermore, there were no detectable HSV-2 or EBV specific RNA or virus-specific antigens in sections of these biopsies. These results provide new lines of evidence for the relationship between CMV and Kaposi's sarcoma. PMID- 6273334 TI - Expression of SV40 T antigen polypeptides in cells biochemically transformed by plasmids containing the herpes simplex virus thymidine kinase gene and the genome of an SV40tsA mutant. AB - To study the expression of SV40 tsA genomes that had been non-selectively introduced into mouse cells, SV40 tsA207 DNA was cleaved with BamH I and ligated to BamH I-cleaved plasmid pAGO DNA, which contains a functional HSV-1 thymidine kinase (TK) gene in the form of 2 kbp Pvu II fragment inserted at the Pvu II site of pBR322. Recombinant plasmids (11-12 kbp) were isolated and amplified in E. coli K12 strain RRI. Restriction nuclease analyses demonstrated that recombinant plasmids pSB15 and pSB10 contained intact SV40 genomes with the polarity of transcription oriented in the same direction (clockwise) or the opposite direction (counterclockwise), respectively, in relation to that of the HSV-1 TK gene. Cla I-cleaved pSB10 and pSB15 DNAs were used to transform LM(TK-) cells to TK+. Serological and disc PAGE analyses showed that clonal lines transformed by these plasmids all expressed the selected marker, HSV-1 TK. Molecular hybridization experiments showed that transformed clonal lines TF pSB10 C7 and TF pSB15 C10 had integrated intact SV40 genomes at one integration site, TF pSB10 C3 had integrated an SV40 genome with a small deletion near the BamH I site, but TF pSB15 Cl had integrated a plasmid from which most of the SV40 nucleotide sequences had been deleted. IF assays with hamster anti-SV40 tumor sera showed that TF pSB10 C7 and TF pSB15 C10 strongly expressed SV40 T antigens in over 90% of the cells, TF pSB10 C3 expressed SV40 T antigens in a minority of the cells, and TF pSB15 C1 did not express SV40 T antigens at all. [35S]-methionine labelling and immunoprecipitation experiments showed that, at 36.5 degrees C: (1) TF pSB10 C7 and TF pSB15 C10 expressed 92K and 20K mol. wt. species of SV40 T antigens and 50-55K cellular protein; (2) expression of all three was reduced in TF pSB10 C3 cells; and (3) TF pSB15 C1 expressed none of the SV40 T antigens, nor did parental LM(TK-) or TF 8-2 transformed cells (which contained the HSV-1 TK gene but not SV40 DNA). At 40 degrees C, labelling of the 50-55K cellular protein was markedly reduced in TF pSB10 C7 and pSB15 C10 cells. The results suggest that SV40 large T antigen (92K) induces and/or stabilizes the 50-55K cellular protein in these mouse cells. PMID- 6273335 TI - Steroids inhibit tumor promoting agent induced Epstein-Barr virus early antigens in Raji cells. AB - Four steroids and one protease inhibitor were evaluated for their effects on 12-O tetradecanoyl phorbol 13-acetate (TPA)-induced and Epstein-Barr virus-induced early antigens (EBV-EA) in Raji cells. Continuous treatment with dexamethasone, prednisolone, hydrocortisone and cortisone inhibited TPA-induced EBV-EA to varying degrees, but the protease inhibitor N-alpha-p-tosyl L-lysine chloromethyl ketone-HCl (TLCK) had no significant effect. None of the agents tested inhibited EBV- induced EA. In addition, the effect of the steroids was reversible since the removal of these agents resulted in recovery of the percentage of EBV-EA-positive cells in TPA-treated cultures. These results were in agreement with the in vivo experiments of other investigators, who demonstrated inhibition of tumor promotion with steroids. Since TLCK failed to inhibit TPA-induced EA, it is unlikely that induction of EA by TPA is the result of production of proteases. PMID- 6273336 TI - Rat mammary carcinoma cells secrete active collagenase and activate latent enzyme in the stroma via plasminogen activator. AB - A spontaneous mammary adenocarcinoma (AC) from an inbred female rat was investigated with regard to secretion of neutral proteases. Cultures of neoplastic epithelial cells derived from the tumour secreted an enzyme that fulfilled the criteria for a specific collagenase. In contrast to cultures of non neoplastic cells, tumour collagenase was present as an active enzyme, since treatment with trypsin or p-aminophenylmercuric acetate (APMA) did not increase activity. The neoplastic cells were also prolific producers of plasminogen activator (PA). Dexamethasone (Dex) (10(-6)M) markedly reduced the levels of both enzymes. Addition of tranexamic acid (TA), an inhibitor of plasmin and of plasminogen activation, did not affect collagenase activity, even at 10(-1)M TA, nor did latent collagenase accumulate. Latent collagenase was secreted in culture by normal fibroblasts from neonatal rat lungs. This latent enzyme was activated by the addition of tumour cell medium plus plasminogen, but this effect was inhibited by the addition of TA. These results demonstrate that the neoplastic cells themselves secrete collagenase as an active enzyme. PA is also secreted, is not involved with tumour collagenase, but is capable, in the presence of plasminogen, of activating latent collagenase produced by the non-neoplastic cells within the tumour or in the surrounding tissue. This tumour possesses potent collagenolytic ability in vitro which may be partly responsible for its rapid invasion in vivo. PMID- 6273337 TI - A geographical study of antibodies to membrane antigens of HSV-2-infected cells and HSV-2-specific antibodies in patients with cervical cancer. AB - Sera was obtained from patients with squamous carcinoma of the cervix from Great Britain (29), Sri Lanka (32), Malawi (27), and Sudan (27), and from controls from all countries except Sudan. Controls were matched for ethnic origin, age and social class. Sera were tested by indirect immunofluorescence (IF) for IgG and IgA antibodies to membrane antigens (MA) and IgA antibodies to VCA of HSV-2 infected cells. Compared with controls, IgA antibodies to MA (IgA anti-MA) were detected more frequently and at higher titres in all groups of patients. However, there was no significant difference in prevalence of these antibodies at titres greater than or equal to 1:4 between Malawian patients and controls, although a significantly higher proportion of patients had IgA anti-MA titres of greater than or equal to 1:16. In contrast, IgA anti-VCA did not distinguish patients from controls. More than 90% of both patients and controls from all countries had IgG antibodies to MA (IgA anti-MA). Malawian patients had a significantly higher geometric mean titre (GMT) of IgG anti-MA than controls and both patients and controls had significantly higher GMTs than their counterparts from other countries. The variation between herpes IgG anti-MA titres in subjects from different countries did not reflect differences in serum immunoglobulin levels and similar variations in titre were not seen in rubella and measles HAI titres. Among the patients there was a geographical variation in the prevalence of HSV-2 specific antibodies detected by ELISA, which varied from 52% among British and Sudanese patients to 73% among Malawian patients. Even when adjustment was made for possible false negative results, there were between 10 and 31% patients without HSV-2-specific antibodies, although only 2 of 103 (1.9%) patients had neither HSV-1 nor HSV-2 antibodies. The association of HSV-2 with cervical carcinoma appeared to vary with age. PMID- 6273338 TI - Retrovirus p30-related antigen in human syncytiotrophoblasts and IgG antibodies in cord-blood sera. AB - Sensitive immunological techniques were used to detect retrovirus markers in human pregnancy. A total of 1,540 human cord-blood sera were tested for retrovirus-reactive IgG antibodies using solid-phase enzyme immunoassay and purified RD 114 virus as antigen. Of these, 118 (7.7%) sera were positive. Blocking assays with specific animal anti-p30 sera, use of control antigen, and electrophoretic protein experiments combined with immunological detection indicated that the human antibodies reacted specifically with the p30 protein. The occurrence of antibodies in cord-blood serum had a highly significant correlation to complications during pregnancy and also correlated to the number of previous abortions and stillbirths. When goat anti-RD 114 p30 serum was used in the peroxidase-anti-peroxidase tissue staining procedure, p30-related antigen was detected in sections of all placental specimens (early and term pregnancies, blighted ova, hydatidiform moles, destructive moles and choriocarcinomas). However, in each case only syncytiotrophoblastic cells were positive. These findings, supplemented with different types of blocking tests, lead us to conclude that retrovirus p30-related antigen is selectively expressed in the highly differentiated syncytiotrophoblasts, which in the normal placenta are directly exposed to maternal blood. It is suggested that retrovirus-reactive antibodies may represent an autoimmune-like immune response to the p30-related syncytiotrophoblast antigen escaping during cellular damage. PMID- 6273339 TI - Liver cell dysplasia in association with hepatocellular carcinoma, cirrhosis and hepatitis B surface antigen in Hong Kong. AB - Liver-cell dysplasia was identified in 60% of 558 cases of cirrhosis with and without hepatocellular carcinoma (HCC) in Chinese coming to necropsy in Hong Kong from 1963 to 1978. A significant correlation was found between liver-cell dysplasia and the identification of hepatitis B surface antigen (HBsAg) in the livers, suggesting that dysplasia may be casually related to hepatitis B virus (HBV). Applying Bayes' theorem to our series of male deaths, we calculated that for Chinese male non-cirrhotics showing liver-cell dysplasia at necropsy, compared with male non-cirrhotics showing no liver-cell dysplasia, the approximate risk factor for HCC is 13:1, whereas for male cirrhotics with liver cell dysplasia, the risk factor for HCC is approximately 2:1. In the presence of dysplasia, the risk for HCC is increased two-fold in males who are also HBsAg positive. Thus, the presence of liver-cell dysplasia in cirrhotic or non cirrhotic livers accompanies a significantly higher risk of developing HCC, especially in the presence of HBsAg. PMID- 6273340 TI - Transformation of rat thyroid epithelial cells by Kirsten murine sarcoma virus. AB - Fischer rat thyroid epithelial cella (FRT) growing continuously in culture were infected with the Kirsten murine sarcoma virus KiMSV(KiMuLV) and found to produce this virus constitutively. Although the morphology of the FRT cells did not change appreciably, the cells became malignant after infection with KiMSV(KiMuLV) as shown by the growth of infected cels in semi-solid media (uninfected FRT cells did not grow) and by the tumorigenicity of infected when injected into syngeneic animals (uninfected FRT cells or FRT cells infected with non-transforming retro viruses were not tumorigenic). The induced tumors morphologically resembled moderately differentiated carcinomas. Two markers of thyroid epithelial differentiation were absent in the original FRT clone and remained unexpressed after transformation. Fully differentiated rat thyroid epithelial cells (FRT-L cells) infected with another strain of the Kirsten murine sarcoma virus, the KiMSV(MolMuLV), were also transformed as demonstrated by the ability also of these cells to grow as carcinomas (after in vitro transformation) in syngeneic animals. Our results clearly demonstrate that the Kirsten murine sarcoma virus can transform in vitro cells of epithelial as well as of fibroblastic origin. PMID- 6273341 TI - Persistent cutaneous herpes simplex infection. AB - A 38-year-old, renal transplant patient developed painful, extensive, chancre like genital ulcerations. The etiological agent was found culturally to be herpes virus type II. The eruption lasted five months and proved recalcitrant to standard therapeutic modalities. On removal of the transplanted kidney and thereby reduction of immunosuppressive agents, the cutaneous viral infection cleared rapidly. The potential severity and persistence of cutaneous herpetic infections in renal transplant patient must be well appreciated or the diagnosis might eaily be missed. PMID- 6273342 TI - Vagal mechanisms in appetite regulation. AB - Though the importance of the vagal nerve for appetitive behavior has been known for a decade, with extensive studies of vagal physiology being performed since the end of the last century, it has not been until recently that major progress has been made in elucidating some of the vagal mechanisms involved in food intake. This short review summarizes methods for studying vagal function, vagal transmission and physiological vagal effects important for appetite regulation normally and in relation to hypothalamic function. Particular emphasis is placed on effects of vagotomy on body weight. PMID- 6273343 TI - Demonstration of an endogenous, highly potent, noncompetitive protein inhibitor(s) of 3H-diazepam binding in bovine brain. PMID- 6273344 TI - Conformation of cyclo-(L-threonine)2 and cyclo-(L-allothreonine)2. A proton and carbon n.m.r study. AB - The diketopiperazines cyclo-(L-Thr)2 and cyclo-(L-allo Thr)2 in water and in dimethyl sulfoxide were studied by proton and carbon-13 nuclear magnetic resonance, and the dominant conformation were deduced from proton-proton and proton-carbon coupling constants. In cyclo-(L-Thr)2 the chi 1 = 60 degrees, hydroxyl over the ring, side chain conformation is favored; this conformation is also favored for cyclo-(L-Ser)2 and cyclo-(L-Ser-D-Ser). However, the important side chain conformation for cyclo-(L-allo Thr)2 is chi 1 = -60 degrees, methyl group over the diketopiperazine ring. The determining factors are apparently steric. The diketopiperazine ring of cyclo-(L-Thr)2 is puckered to hold the side chains more nearly axial than is that of cyclo-(L-allo Thr)2. although the degree of ring folding is probably not large. PMID- 6273345 TI - Modification of lysine 69 reactivity in bovine growth hormone by carbamylation of its N-terminal group. AB - Bovine growth hormone was carbamylated under conditions that assure full reaction of the N-terminal residue. Approximately 28% of lysine 179 and 7% of lysine 143 were also carbamylated. The modified hormone retained an important growth promoting activity and was as effective as the native hormone in competition assays in vivo for the receptors in rat liver. However, a change in its conformation must occur since lysine 69, which is resistant to trinitrophenylation in the native hormone, reacted easily and under mild conditions, in the carbamylated protein, The growth promoting activity and binding properties of the carbamylated and trinitrophenylated hormone were practically nil. PMID- 6273346 TI - Studies of peptide antibiotics. XLI. Syntheses of 6-L-valine-tyrocidine A and 7-L ornithine-tyrocidine A. AB - Tyrocidine A (TA) is an antibiotic cyclic decapeptide with the sequence of cyclo (-L-Val1-L-Orn2-L-Leu3-D-Phe4-L-Pro5-L-Phe6-D-Phe7-L-Asn8-L-Gln9-L-Tyr10-). Gramicidin S (GS) regarded as a homolog of TA is also a cyclic decapeptide with the sequence of cyclo (-L-Val1-L-Orn2-L-Leu3-D-Phe4-L-Pro5-L-Val5-L-Orn7-L-Leu8-D Phe9-L-Pro10-). GS shows higher antibacterial activity, whereas TA exhibits inhibitory activity on the biosynthesis of RNA. Two analogs of TA, [L-Val6]-TA (12a) and [L-Orn7]-TA (12b), were synthesized by the conventional method in order to study the interrelationships between the two related antibiotics TA and GS. Antibacterial activities of 12a and TA are nearly the same, but the activity of 12b is significantly lower. The optical rotatory dispersion spectra of 12a, 12b, and TA showed a trough at 233 nm region; the troughs of 12a and TA are nearly the same in depth, but the trough of 12b is shallower. Relationships between structure and activity of 12a and 12b compared with TA and GS were discussed. PMID- 6273347 TI - Comparison of the structural features of corticotropin required for stimulation of steroidogenesis and cAMP production in rat and rabbit adrenocortical cells. AB - The steroidogenic activities of ACTH, alpha-MSH, beta-MSH as well as analogs of the hormones have been compared in rat and rabbit adrenocortical cells. ACTH is equally active in both species and the melanotropins have very low steroidogenic potency in either species. The steroidogenic potencies of the peptide analogs are strikingly similar in the two species, suggesting that the structural requirements for eliciting steroidogenesis are the same in rat and rabbit adrenocortical cells. The analog NPS-ACTH has low, comparable steroidogenic activity in both species. NPS-ACTH is a potent antagonist of ACTH-induced cAMP production in rat adrenocortical cells but acts as a weak partial agonist in rabbit adrenocortical cells. These results suggest that steroidogenesis may be mediated by receptors different from those involved in the cAMP response observed at supraphysiological concentrations of ACTH. PMID- 6273348 TI - 1H N.M.R. studies of protected alpha-aminoisobutyric acid containing peptides. Chemical shift nonequivalence of benzyloxycarbonyl methylene protons. AB - The benzylic methylene protons in a large number of benzyloxycarbonyl alpha aminoisobutyric acid (Z-Aib) containing peptides, show chemical shift nonequivalence. The magnitude of the geminal nonequivalence is correlated with the involvement of the urethane carbonyl group, in an intramolecular hydrogen bond. Studies of the model compounds Z-Aib-Aib-Ala-NHMe, and Z-Aib-Aib-Aib-Pro OMe clearly establish the presence of intramolecular hydrogen bonds, involving the urethane CO group. In both compounds marked anisochrony of the benzylic methylene protons is demonstrated. In Z-Aib-Aib-Pro-OMe, where a 4 leads to 1 hydrogen bonded beta-turn is not possible, the benzylic-CH2-protons appear as a singlet in CDCl3 and have a very small chemical shift difference in (CD3)2SO. The observation of such nonequivalence is of value in establishing whether the amino terminal Aib-Pro beta-turn is retained in large peptide-fragments of alamethicin. PMID- 6273349 TI - Clostripain-catalyzed re-formation of a peptide bond in a cytochrome C fragment complex. AB - Enzymatically-catalyzed condensation of cytochrome c fragments, ferrous heme fragment (1-38) and apofragment (39-104), has allowed the back-conversion of cytochrome c complex to native cytochrome c. The conversion was accomplished in 90% (v/v) glycerol, a solvent which has been shown to decrease the ionization of the terminal alpha-carboxyl group liberated during hydrolysis of a peptide bond. The effect on the pK is probably the main reason the thermodynamic obstacle to re synthesis is minimized. A 30% conversion to cytochrome c was obtained. The cytochrome c product was distinguished from the non-covalent complex and separated fragments by molecular weight analysis with sodium dodecyl sulfate polyacrylamide gel electrophoresis, by elution from Sephadex G-50 and sulfopropyl Sephadex in the presence of denaturant, by amino acid analysis of the product purified under complex-dissociation conditions, and by spectral analysis of the absorption bands of the heme. This method provides an opportunity to study the covalent rather than the complex form of cytochrome c analogs. PMID- 6273350 TI - Human somatotropin. Noncovalent interaction of two thrombin fragments restores receptor-binding activity. PMID- 6273351 TI - Beta-endorphin. Vasodilating effect on the microcirculatory system of hamster cheek pouch. AB - beta-Endorphin, enkephalins and morphine dilated in vivo the arteriole of the microcirculatory system in hamster cheek pouch. beta-Endorphin had the highest potency and morphine the lowest. The effect was dose dependent and blocked competitively by the opiate antagonist, naloxone. The result demonstrated an effect of the opiates on the arteriole of the cutaneous circulation. This effect probably plays an important role in thermoregulation. PMID- 6273353 TI - [Pro and con: small cell bronchial cancer - surgery or chemotherapy as the primary treatment. Arguments for primary chemotherapy]. PMID- 6273352 TI - Illumination dependent hyperpolarization of the photoreceptor outer segment membrane (role of calcium, cyclic GMP and calmodulin). AB - Calmodulin has been isolated from the outer segment of toad (Bufo marinus) photoreceptors. Bioelectric processes have been recorded from the same preparation during various experimental conditions. The results led to the conclusion that during illumination the rotation of the chemoreceptors opens the Ca channels, and free Ca is versed into the intermembrane space of the outer segment. Changes of this free Ca concentrations depend on fast influx and slower removal (mainly dependent on Ca binding on calmodulin and back-diffusion into the disc membrane). The changes of the free Ca concentrations generate the changing hyperpolarization, a Ca-dependent process of first phases of encoding the photic stimulus. The effectiveness of Ca to generate the oscillating hyperpolarization depends on optimum state of bioelectric potentials. This potential seems to be regulated by the effective decrease of cGMP concentrations by the increased activity of the cGMP phosphodiesterase during illumination (a sudden and gross regulation). A finer regulation is exerted by Ca during its release and removal (e.g., Ca-dependent decrease of Na channels, decreased release of cGMP by Ca dependent inhibition of guanylate cyclase activity and increased phosphodiesterase activity by both free Ca and Ca-bound calmodulin). PMID- 6273354 TI - [Pro and con: small cell bronchial cancer - surgery or chemotherapy as the primary treatment. Arguments for selective primary resection]. PMID- 6273355 TI - Evaluation of the antiherpetic activity of 2'-fluoro-5-iodo-ara-C in rabbit eyes and cell cultures. AB - 2-Fluoro-5-iodo-ara-C (FIAC), a new and potent drug, was tested for antiviral activity against several strains of herpes simplex virus (HSV), types 1 and 2. Effective dose-50% (ED-50) determinations for FIAC ranged from 0.023 to 0.51 muM for HSV-2. FIAC-treated cells did not exhibit any toxicity until the drug concentration was increased 2000-fold above the ED-50 level. Ocular herpetic keratitis in New Zealand white rabbits was treated with 1.0%, 0.1%, 0.01% FIAC beginning 3 days after inoculation wit HSV-1 (McKrae strain). Topical chemotherapy was administered five times per day for 7 consecutive days. After 4 days of treatment, corneal epithelial involvement, conjunctivitis, iritis, and clouding were not detectable in eyes receiving 1.0% FIAC. Toxic reactions were not observed in rabbit eyes treated with FIAC drug. HSV was not prevented from spreading into the central nervous system when topical FIAC therapy was initiated on day 3 after inoculation. PMID- 6273356 TI - Quantitation of herpes simplex virus in vitro and in preocular tear film. AB - Current methods for virus isolation from preocular tear film are not quantitative. This report presents a sensitive method for detection and quantitation of herpes simplex virus type 1 in unknown samples of small size (less than 50 microliters). Serial tenfold dilutions of stock virus were inoculated onto 16 mm vero cell monolayers, which were monitored for the development of cytopathologic evidence (CPE) for presence of virus and were assigned a severity grade. Standard curves were developed on the basis of time interval between inoculation, recognition of CPE, and CPE score. These curves were used to predict virus titers in unknown samples. This method for virus isolation is simple, efficient, and consistently sensitive to virus titers of less than 10 plaque forming units. PMID- 6273357 TI - Cannabis and health - a review. PMID- 6273358 TI - The pathologic basis for breast cancer management. AB - The pathologist's report on breast cancers should include information that assists the clinician to select therapy, estimate prognosis, and evaluate the efficacy of therapy. Of great significance are the size and contour of the primary cancer, histologic type, differentiation, vascular invasion, the total number of lymph nodes extirpated, the number of metastases, and whether the metastases are micrometastases (less than 2 mm) or macrometastases. PMID- 6273359 TI - Cystosarcoma phyllodes. AB - A series of 27 cystosarcoma phyllodes diagnosed in 23 patients is presented. Treatment consisted of wide local excision with a rim of normal tissue for the histologically benign-appearing tumors, and simple mastectomy for the histologically malignant-appearing tumors. Of the three tumors with a malignant appearance, one metastasized distantly and the patient died; the other two patients were apparently cured. Of the 24 benign-appearing tumors, 3 recurred locally and were cured by reexcision. None behaved in a malignant fashion. There was one case of bilateral cystosarcoma phyllodes, and one tumor was associated with carcinoma of the breast. The local recurrence rate of 11% (3 out of 27) and mortality rate of 4% (1 out of 23) appears to justify this therapeutic approach. The precise indications for mastectomy in this disease are still unclear. PMID- 6273360 TI - Maintenance of prolactin receptors in human breast cancer. AB - Breast tissue specimens of 110 women with various stages of breast cancer were tested in vitro to determine their specific binding sites for human prolactin. In contrast to the case of steroid receptors, binding sites for prolactin were found in the vast majority of breast cancer tissue. Distribution profiles giving amount of prolactin receptor and their affinity coefficients were found to be similar in the tissues of women whose ages, hormonal status, or stage of breast cancer varied. These findings show that in contrast to steroid receptors, human breast cancer tissue maintains binding sites for prolactin. The findings also indicate that there may be a higher dependency of breast cancer on prolactin than on steroids. Clinical trials must be carried out to determine the role of "positive" prolactin receptors in prognosis and prediction of response to future hormone therapy. PMID- 6273361 TI - The elevation of mouse thymus cell cyclic adenosine monophosphate (cAMP) by lipopolysaccharide. AB - The ability of certain lipopolysaccharide (LPS) preparations to elevate cyclic adenosine monophosphate (cAMP) in mouse thymus cells in the presence of Ro 20 1724, 4-(3-butoxyl 4-methoxybenzyl)-2-imidazolidinone, was not related to the source of supply, bacterial strain, or method of extraction. Under the same conditions adenosine is a potent stimulator of thymus cell cAMP and is, of course, blocked by the further addition of theophylline. When theophylline was added to the LPS preparations with Ro 20-1724, the cAMP production was also blocked. These studies suggested that the observed stimulation of cAMP by LPS preparations was due to adenosine and/or its nucleotides present as contaminants. PMID- 6273362 TI - [Consideration of biorhythms in systemic dermatotherapy with corticosteroids]. AB - Rhythmic, in particular circadian, variations of body functions can be found in, or at least supposed for, almost all functions of the skin. The periodicity of physiological properties, some of which occur phase shifted to each other, represents the main reason for circadian differences of both the bioavailability and susceptibility to equal doses of drugs given at different day times. With regard to the well known circadian rhythm of ACTH and cortisol secretion, which can be suppressed by high dose administration of corticoids, every systemic corticoid treatment should, as a rule, be restricted to the morning hours, i.e. during the acrophase of endogenous cortisol secretion. Since after systemic or even topic corticoid therapy the rhythm of ACTH-cortisol feedback system may normalize with marked delay, skin reactivity tests as well as immunodiagnostic in vitro measures should be done just after appropriate poststeroid time lapse to avoid erroneous results. Some alteration of the circadian cortisol rhythm without foregoing corticoid treatment has also been found in cases of generalized atopic dermatitis as well as progressive systemic sclerosis of acrosclerotic type. PMID- 6273363 TI - [Virus papilloma in humans]. PMID- 6273365 TI - Florid dermal cylindroma (turban tumor). AB - A florid case of dermal cylindroma is presented as representative of the natural history and usual clinical findings in a patient afflicted by this tumorous growth. Although histologically and clinically benign, dermal cylindroma can result in considerable disfigurement of the patient. Treatment varies from isolated primary resection to serial resections with subsequent electrocautery or cryosurgery for small recurrences. Total scalping with skin grafting is reserved for only the most severe cases where normal integument has been completely involved by the disease process. PMID- 6273364 TI - Nuclear medicine in diagnosis and treatment of diseases of the head and neck. I. Salivary and parathyroid gland disease and one identification and staging of head and neck tumors. AB - The advent of both improved imaging systems and new radioactive agents has increased the effectiveness of nuclear medicine in diagnosing and treating diseases of the head and neck. In this first in a series of two articles, the role of nuclear medicine is discussed in the evaluation of diseases of the salivary and parathyroid glands, and in the identification and staging of head and neck tumors. Radionuclide studies of the salivary glands are useful in the identification of tumors and the evaluation of gland function. Such studies are a valuable adjunct in the diagnosis of Sjogren's syndrome and of acute and chronic inflammatory disease. Radionuclide imaging also has been helpful in the detection of adenomata and hyperplasia of the parathyroid glands and often complements ultrasonography localization procedures. The advent of gallium-67 imaging has improved the staging of head and neck tumors. PMID- 6273366 TI - Pituitary function in 8 patients with familial pituitary dwarfism. AB - Pituitary function was evaluated in four pairs of familial pituitary dwarfs (two sisters, 28 and 20 years old, two brothers 15 and 10 years old, one girl 19 years old and her brother 7 years old, and one girl 9 years old and her brother 3 6/12 years old), who all proved to have multiple pituitary hormone deficiency (MPHD), in order to find out whether a distinct intrafamilial or interfamilial pattern of deficiency existed. The response of FSH and LH to LRH, of TSH to TRH, of growth hormone (GH) to L-dopa, and of prolactin to sulpiride administration was studied. Basal levels of ACTH and cortisol (F) were also measured on two consecutive days. Basal FSH, LH and GH concentrations were below the sensitivity of the assay in all patients (except one who had measurable levels of LH) and showed no change after the appropriate stimulation. Basal TSH was normal in 6 patients and slightly increased in two members of the same family. Both these patients presented an exaggerated response to TRH. Three of the remainder patients showed a small and the other three no response to TRH. Basal prolactin values ranged within the normal limits, but prolactin was not increased during the sulpiride provocative rest. Mean basal ACTH was normal in 5 and low in 3 patients. Mean cortisol levels were normal in 5, near the lower normal limit in 2, and low in one patient. This patient had also a low ACTH concentration. It is concluded that the lack of a common pattern of hormone deficiency in all families indicates a variability in the expression of the hypothalamo-pituitary defect, which suggests that the familial MPHD dwarfism constitutes a genetically and pathogenetically heterogenous group. PMID- 6273367 TI - Localization of recently characterized membrane transport adenosine triphosphatases. PMID- 6273369 TI - Familial chemodectoma. PMID- 6273368 TI - 5'-Nucleotidase activity in lymphocytes. AB - Some characteristics of lymphocyte 5'-nucleotidase are reviewed. The optimal conditions for the cytochemical localization of 5'-nucleotidase (AMPase) in the mouse lymphocyte have been established. Quantitative monitoring of the effects of fixation and the components of the cytochemical medium showed that cytochemistry can be performed under conditions that do not lead to loss of AMPase activity. The cytochemical reaction product was seen only on the surface of a proportion of splenic lymphocytes, regardless of the fixative used. The splenic cell population included a distinct population of lymphocytes with readily demonstrable AMPase activity and another population with no cytochemically demonstrable AMPase activity. The number of positive cells varied, but was usually between 20 and 30% when cells were fixed in glutaraldehyde. Lymphocytes purified from thymus were always negative for cytochemically demonstrable AMPase activity. Biochemically, it was shown that the AMPase activity of spleen lymphocytes is more than six times higher than that of thymus lymphocytes. 5'-Nucleotidase activity in normal and abnormal lymphocytes is discussed in the light of the latest findings. The possible function of 5'-nucleotidase in lymphocytes is discussed. PMID- 6273370 TI - Effect of hepatic irradiation on the toxicity and pharmacokinetics of adriamycin in children. PMID- 6273371 TI - Client assessment: an integrated model. PMID- 6273373 TI - The use of case studies in bioethics courses. PMID- 6273372 TI - The effect of freedom on the learning of gifted nursing students. PMID- 6273374 TI - The West Virginia Plan: a portrait of curriculum development. PMID- 6273375 TI - Faculty and student perceptions of effective clinical teachers. AB - The following study has been conducted to discover the characteristics of an effective clinical teacher as described by faculty and students in a selected baccalaureate school of nursing. Through determination of these characteristics, clinical teachers can improve their teaching methodology and provide a more effective learning environment for the learner. The identification of these characteristics also has implications for graduate nursing education, as clinical teachers are being prepared in this arena to enter the teaching environment. Administrators should consider utilization of these characteristics for teacher evaluation purposes. Nursing educators must attempt to bridge the gap between what educators and students perceive as characteristics of the effective clinical teacher. The ultimate goal to be achieved from identification of effective clinical teacher characteristics, as determined in this study, is improvement in clinical teaching. PMID- 6273377 TI - Atomic absorption spectrometric determination of tin in canned foods using nitric acid-hydrochloric acid digestion and nitrous oxide-acetylene flame. AB - Foods (5-40 g) are digested sequentially with HNO3 and HCI, diluted to 100 mL, and filtered, and tin is determined by atomic absorption using an N2O-C2H2 flame. Synthetic standards in 10% HCl are used. A positive interference by potassium is compensated by adding 100 micrograms potassium/mL to both standards and samples. Recoveries for 17 foods spiked with tin at levels between 40 and 500 micrograms/g ranged from 85 to 110% with a mean of 101%. Results agreed well with a solvent extraction atomic absorption method and a direct X-ray fluorescence method. PMID- 6273376 TI - Interaction of beta-lactamase of Streptomyces cacaoi. II. CP-45,899, izumenolide and cephamycins. AB - Inhibition of a beta-lactamase of Streptomyces cacaoi by CP-45,899, izumenolide and cephamycins was investigated and compared with that of a beta-lactamase of Bacillus cereus. S. cacaoi enzyme could not hydrolyze CP-45,899. Instead, hydrolysis of benzylpenicillin by the enzyme was inhibited in the presence of CP 45,899. Although inhibition increased gradually with time, the inhibition line produced by CP-45,899 with time less curved than that produced by clavulanic acid and PS-5. Furthermore, preincubation of S. cacaoi beta-lactamase with CP-45,899 for up to 120 seconds did not obviously affect the degree of inhibition. When the concentration was lowered, it behaved as a competitive inhibitor, a Ki value being 6.2 X 10(-7) M. Izumenolide, on the other hand, did not inhibit the enzyme activity of S. cacaoi beta-lactamase at 1.28 X 10(-4) M, although it inhibited B. cereus enzyme slightly in a competitive manner. Oganomycins were inert to the both beta-lactamases.U PMID- 6273378 TI - Comparison of dietary fiber methods for foods. AB - In order to evaluate several proposed dietary fiber methods, 12 food samples, representing different food classes, were analyzed by (1) neutral and acid detergent fiber methods (NDF, ADF); (2) NDF with enzyme modification (ENDF); (3) a 2-fraction enzyme method for soluble, insoluble, and total dietary fiber, proposed by Furda (SDF, IDF, TDF); (+) a 1-fraction enzyme method for total dietary fiber proposed by Hellendoorn (TDF). Foods included cereals, fruits, vegetables, pectin, locust bean gum, and soybean polysaccharides. Results show that TDF by Furda and Hellendoorn methods agree reasonably well with literature values by the Southgate method, but ENDF is consistently lower; that ENDF and IDF (Furda method) agree reasonably well; that except for corn corn bran fiber (insoluble) and pectin and locus bean fiber (soluble), all materials have significant fractions of both soluble and insoluble fiber. The Furda method was used on numerous food and ingredient samples and was found to be practical and informative and to have acceptable precision (RSD values of 2.65-7.05%). It is suggested that the Furda (or similar) method be given consideration for the analysis of foods for dietary fiber. PMID- 6273379 TI - Accumulation of 99mTc-pyrophosphate in malignant pericardial effusion. PMID- 6273380 TI - Isolation and characterization of an Escherichia coli mutant affected in the regulation of adenylate cyclase. AB - A mutant, cyaR1, affecting regulation of adenylate cyclase expression or activity is described. It was obtained as a thermoresistant revertant of a strain harboring a thermosensitive transcription termination factor, rho (rho-15). This mutant failed to synthesize adenosine 3',5'-phosphate and exhibited a carbohydrate-negative phenotype. A secondary mutation at the crp locus (crpC) restored the ability of the mutant to synthesize adenosine 3',5'-phosphate, enabled the expression of catabolite-sensitive operons, and conferred on the strain an extreme sensitivity to catabolite repression. In addition, we showed that the crpC mutation restored the pleiotropic carbohydrate-positive phenotype even in a delta cya background. We interpret this to mean that the adenosine 3',5'-phosphate receptor protein regulates negatively either the activity or synthesis of adenylate cyclase and that the cyaR1 mutation is either in a regulatory protein or a regulatory site of adenylate cyclase. PMID- 6273381 TI - Plasmid RP4 specifies a deoxyribonucleic acid primase involved in its conjugal transfer and maintenance. AB - We surveyed plasmids representative of most incompatibility groups for their conferred deoxyribonucleic acid (DNA) primase activity. RP4 (IncP) was one of the few with such activity although, unlike the derepressed IncIalpha plasmids (which also specify a primase), it did not suppress the dnaG mutation. Using deletion and Tn7 derivatives of RP4, we located the presumed primase structural gene (pri) in the 37- to 42-kilobase region. Tn7 insertions in the adjacent Tra1 region also reduced or caused overproduction of primase. We purified the RP4 primase to a single polypeptide of molecular weight 118,000. It is an anisometric molecule and functions as a monomer, initiating complementary strand synthesis on phi X174 DNA in Escherichia coli dnaG cell extracts in the presence of ribonucleotide triphosphates and rifampin. It is immunologically unrelated to either the E. coli dnaG or the IncIalpha plasmid-specified DNA primases. RP4 pri mutants conjugated with a lower efficiency into some bacterial species, including Salmonella typhimurium. Back-transfer experiments showed that this effect was recipient specific. There was also a comparable reduction in mobilization efficiency of R300B by RP4 pri into such recipients. Loss of RP4 primase led to detectable plasmid instability. The RP4-specified primase therefore seems to serve two functions: the single DNA strand transferred during conjugation is primed by it in the recipient cell, and it appears to be necessary for the efficient priming of discontinuous plasmid DNA replication despite the presence of the chromosomal priming system. PMID- 6273382 TI - Molecular nature of a plasmid specifying beta-lactamase production in Haemophilus ducreyi. AB - We characterized pJB1, the plasmid previously reported to mediate beta-lactamase production in Haemophilus ducreyi. We studied its relationship to pMR0360 and RSF0885, the plasmids responsible for beta-lactamase production in Neisseria gonorrhoeae and Haemophilus parainfluenzae, respectively. Although pJB1 was maintained as a multicopy pool in Escherichia coli, it was not stably maintained in the absence of antibiotic selection. Electron microscope heteroduplex studies showed that it carried 100% of the transposable ampicillin resistance sequence TnA. This sequence was transposed to plasmid pUB307 at a low rate. Heteroduplexes between pMR0360 and pJB1 showed that they contained 3.3 megadaltons of homologous sequences. Two sets of nonhomologous sequences, one a TnA sequence and the other a non-TnA sequence, took the form of insertion loops. For plasmids pMR0360 and RSF0885, previously shown to be highly related, the nonhomologous sequences took the form of a substitution loop. We concluded that all three plasmids shared major portions of their sequences but differed in discrete segments. pJB1 was the first such plasmid to have a physically and functionally intact TnA sequence. PMID- 6273383 TI - Plasmid-determined immunity of Escherichia coli K-12 to colicin Ia Is mediated by a plasmid-encoded membrane protein. AB - The colicin Ia structural (cia) and immunity (iia) genes of plasmid pColIa-CA53 have been cloned into the cloning vector pBR322. These two genes are closely linked, and both of them can be isolated on a deoxyribonucleic acid fragment approximately 4,800 base pairs long. An analysis of the polypeptides synthesized in ultraviolet-irradiated cells containing these cloned genes led to the conclusion that the iia gene product is a polypeptide with a molecular weight of approximately 14,500. Insertion of transposon Tn5 into the iia gene led to a concomitant loss of the immune phenotype and the ability to produce this protein. Fractionation of ultraviolet-irradiated cells harboring a plasmid carrying the iia gene showed that the immunity protein is a component of the inner (cytoplasmic) membrane. Furthermore, the mechanism of immunity to colicin Ia appears to operate at the level of the cytoplasmic membrane. This conclusion is based on our finding that membrane vesicles prepared from colicin Ia-immune cells could be depolarized by colicins E1 and Ib but not by colicin Ia. PMID- 6273384 TI - Homology between the invertible deoxyribonucleic acid sequence that controls flagellar-phase variation in Salmonella sp. and deoxyribonucleic acid sequences in other organisms. AB - The invertible deoxyribonucleic acid (DNA) segment cloned from Salmonella sp. was radioactively labeled and used as a probe to search for homologous sequences by Southern hybridization. Only one copy of the invertible segment could be found on the Salmonella sp. genome. Partial sequence homology with the invertible region was detected in bacteriophage Mu and P1 DNA by low-stringency hybridization. Under these conditions, no homology was detected with Escherichia coli DNA. A strain of Salmonella sp. defective in phase variation carrying the vH2- allele was also analyzed by DNA-DNA hybridization. The results show that there is sequence divergence between diphasic and vH2- strains within the invertible sequence. PMID- 6273386 TI - Electron transport system of an aerobic carbon monoxide-oxidizing bacterium. AB - Experiments with crude extracts of Pseudomonas carboxydohydrogena revealed that a quinone is necessary for CO oxidation, and that cytochromes of the a, b, and c types and functional terminal oxidase(s) are found in cells grown on CO as the sole source of carbon and energy. CO dehydrogenase delivers electrons to the electron transport system at the level of quinone, and nicotinamide adenine dinucleotide (phosphate) is not involved in CO oxidation. PMID- 6273385 TI - Virulence-associated plasmids from Yersinia enterocolitica and Yersinia pestis. AB - A 44-megadalton plasmid associated with virulence and Ca2+ dependence from Yersinia enterocolitica 8081 was compared at the molecular level with a 47 megadalton plasmid associated with Ca2+ dependence from Yersinia pestis EV76. The plasmids were found to share 55% deoxyribonucleic acid sequence homology distributed over approximately 80% of the plasmid genomes. One region in which the plasmids differed was found to contain sequences concerned with essential plasmid functions. Forty-five mutants of Y. pestis were isolated which had spontaneously acquired the ability to grow on calcium-free medium (Ca2+ independence). Of these mutants, 21 were cured of their 47-megadalton plasmid, whereas the remaining had either suffered a major deletion of the plasmid or had a 2.2-kilobase insertion located in one of two adjacent BamHI restriction fragments encompassing approximately 9 kilobases. The inserted sequence was found at numerous sites on the Y. pestis chromosome and on all three plasmids in the strain and may represent a Y. pestis insertion sequence element. PMID- 6273387 TI - The subunit composition of pea cytochrome c oxidase. AB - Cytochrome c oxidase was purified from pea shoots in a form containing more than 12 nmol of heme a per mg protein, but rapid inactivation took place during purification. On slab polyacrylamide concentration gradient gel electrophoresis of a partially purified preparation, there were three activity-bands corresponding to main protein bands. The activity-bands, as well as the most purified preparation, contained five polypeptides of different molecular weights (39,000, 33,000, 28,500, 16,500, and 8,000-6,000) as shown by sodium dodecylsulfate-urea polyacrylamide gel electrophoresis. However, an immunoprecipitate from the partially purified preparation with antibody against the most purified preparation contained two additional polypeptides with molecular weights of 13,000 and 10,000. Pea cytochrome c oxidase resembled the sweet potato enzyme with respect to immunological properties and absorption spectra as well as the subunit composition. We propose that higher plant cytochrome c oxidase is composed of five subunits of different molecular weights and is associated weakly with two low-molecular-weight polypeptides in the mitochondrial inner membrane. PMID- 6273388 TI - The interactions of lysophosphatidylcholine with protein-containing liposomes. AB - The effect of exogenous lysophosphatidylcholine on the membranes of liposomes containing protein has been studied. Lysophosphatidylcholine severely damaged liposomes prepared in the presence of membrane proteins such as glycophorin and "band 3" protein of human erythrocytes. Some basic proteins such as cytochrome c, lysozyme and polylysine also could sensitize liposomes to lysophosphatidylcholine. As described in previous papers (Inoue, K., et al. (1974) Biochim. Biophys. Acta 363, 361-372; Utsumi, H., et al. (1978) Biochemistry 17, 1990-1996), large multilamellar liposomes without protein were affected by lysophosphatidylcholine only under certain conditions where a phase boundary could exist. Sonicated liposomes without protein were almost completely insensitive to lysophosphatidylcholine. Liposomes prepared by the cholate dialysis method were also insensitive to lysophosphatidylcholine, irrespective of the incubation temperature. It is known that natural membranes such as membranes of erythrocytes and of other mammalian cells are rather sensitive to lysophosphatidylcholine. The difference observed between natural membrane and liposomal membrane seems to be removed by the introduction of proteins into lipid bilayers. The mode of interaction of lysophosphatidylcholine with membranes of liposomes containing protein is discussed. PMID- 6273389 TI - Isolation and immunochemical localization of collagenase in mouse peritoneal macrophages. AB - Collagenase was isolated from the culture medium of thioglycollate-stimulated mouse peritoneal exudate macrophages. The macrophage collagenase activity was inhibited by goat anti-mouse bone collagenase antibody, indicating that macrophage collagenase immunologically cross-reacts with mouse bone collagenase. The enzyme was localized in mouse peritoneal macrophages by indirect immunofluorescent antibody technique. Distinct granular fluorescence was observed intracellularly in most thioglycollate-stimulated macrophages, whereas slight or no fluorescence was observed in non-stimulated control macrophages. PMID- 6273390 TI - Reconstitution of urea-dissociated subunits of a pig heart phosphoprotein phosphatase. AB - Pig heart phosphoprotein phosphatase [phosphoprotein phosphophydrolase, EC 3.1.3.16] of Mr 224,000 was dissociated by gel-filtration on Sephacryl S-300, into an active subunit (alpha subunit) of Mr 31,000 and inactive subunits of higher molecular weight in the presence of 6 M urea. After the removal of urea, these subunits reassociated, forming two enzyme forms of Mr 237,000 (Form 1) and Mr 123,000 (Form 2). Form 2 was produced by association of the alpha subunit with an inactive subunit (beta subunit) of Mr 80,000, while Form 1 was formed by combination of the alpha subunit with a complex of inactive subunits which was eluted from a Sephadex G-150 column in fractions of molecular weight range greater than 80,000. The dissociation and reassociation of the subunits of Form 1 by the same urea method produced not only Form 1, but also significant amounts of Form 2, indicating that the inactive subunits of Form 1 were a complex of the beta subunit with another inactive subunit(s). The molecular parameters and other properties of Form 1 were very close to those of the original enzyme. By the conversion of Form 1 to Form 2, the activities of Form 1 towards phosphorylase a and glycogen synthetase b were enhanced 2-3 fold with no significant change in activity towards P-H1 histone or in response to the stimulatory effect of Mg(CH3COO)2 on the dephosphorylation of P-H2B histone. However, removal of the beta subunit from From 2 resulted in strong suppression of activity towards P-H1 histone and response to the salt effect with lesser effects on the activities of Form 2 towards phosphorylase a and glycogen synthase b. PMID- 6273391 TI - Molecular structures of packageable ColE1 hybrids and the generation of deletion mutants from one of the hybrids. AB - ColE1 derivatives carrying cohesive end sites of lambda phage genome (= cos lambda) can be packaged within lambda phage particles. The DNA structure of the prototype ColE1-cos lambda derivative named pKY2257 was studied because of its potential usefulness in various fields in molecular biology. pKY2257, which carries an intact galactose operon of E. coli, is a convenient replicon to detect Tn3 translocation. It was found that one of the PK2257::Tn3 derivatives, pKY2113, generated various small plasmids in E. coli. The molecular structures of some of these deletion mutants were compared with each other and with those of parental plasmid DNAs by heteroduplex analysis and restriction enzyme digestion. A possible mechanism, which seems to be unique to this kind of deletions, is discussed on the basis of the present results. PMID- 6273392 TI - The effect of progressive starvation in vivo on amino acid transport and rubidium uptake in monolayer cultures of rat hepatocytes. PMID- 6273393 TI - Histone-dependent ADP-ribosylation of low molecular nucleotide by poly(ADP ribose) polymerase. PMID- 6273394 TI - Studies on algal cytochromes. III. Amino acid sequence of cytochrome c-553 from a brown alga, Petalonia fascia. AB - The amino acid sequence of a photosynthetic cytochrome c-553 isolated from a brown alga, Petalonia fascia was determined by BrCN fragmentation and a solid phase Edman degradation. The cytochrome contains 85 amino acid residues, giving a molecular weight of 9,803. The complete amino acid sequence is as follows: Val Asp-Ile-Asn-Asn-Gly-Glu-Ser-Val-Phe-Thr-Ala-Asn-Cys-Ser-Ala-Cys-His-Ala-Gly -Gly Asn-Asn-Val-Ile-Met-Pro-Glu-Lys-Thr-Leu-Lys-Lys-Asp-Ala-Leu-Glu-Glu-Asn-Gl u-Met Asn-Asn-Ile-Lys-Ser-Ile-Thr-Tyr-Gln-Val-Thr-Asn-Gly-Lys-Asn-Ala-Met-Pro-A la-Phe Gly-Gly-Arg-Leu-Ser-Glu-Thr-Asp-Ile-Glu-Asp-Val-Ala-Asn-Phe-Val-Ile-Ser-Gln-Ser Gln-Lys-Gly-Trp. The highest homology was found between the sequences of cytochromes c-553 of P. fascia and Alaria esculenta, the next between those of P. fascia and Porphyria tenera. PMID- 6273395 TI - Sodium ion discharge from pig kidney Na+, K+-ATPase Na+-dependency of the E1P-E2P equilibrium in the absence of KCl. AB - The two phosphoenzymes (E1P and E2P) of Na+,K+-ATPase were measured as ADP sensitive and K+-sensitive fractions. The sum of these fractions was nearly 1 in the range of 50 to 1,200 mM NaCl. The effects of Na+ on the levels of E1P and E2P, on the rate constant of E2P leads to E1P transition (k2), on the rate constant of E2P dephosphorylation (k3), on the rate constant of E1P leads to E2P transition (k1) and on the apparent equilibrium constant between E1P and E2P (Kapp) were examined. k1 was found to decrease with increasing Na+ concentration, whereas k2 increased. Kapp was found to be directly proportional to the third power of Na+ concentration. k3 increased with increasing Na+ concentration and saturated at about 1 M NaCl. These results are consistent with a simple model in which ATP hydrolysis occurs through effectively only two phosphoenzyme intermediates in the absence of K+ and three sodium ions are discharged cooperatively from the enzyme during the E1P leads to E2P conversion. PMID- 6273396 TI - The partially reduced species present in purified cytochrome oxidase from baker's yeast is cytochrome a. AB - Cytochrome oxidase purified from baker's yeast submitochondrial particles is found to exist in a partially reduced state in the resting enzyme. Studies utilizing optical and EPR spectroscopy indicate that the "inactive" fraction contains a reduced low-spin heme, cytochrome a possibly indicating a block of electron transfer from cytochrome a to cytochrome a 3. There is no apparent reduction of either the EPR-detectable copper or the species associated with the 830 nm band. Oxidative titrations of the resting-state yeast cytochrome oxidase indicate that the reduction potential of the species titrating is higher than that of ferricyanide. This "inactive" cytochrome oxidase is not the result of the isolation procedure, but seems to represent a species which is present in the intact yeast. PMID- 6273397 TI - Similarities between the transferrin receptor proteins on human reticulocytes and human placentae. AB - The transferrin receptor of the human reticulocyte was isolated by two different immunoaffinity procedures. These included indirect immunoprecipitation with a transferrin/anti-transferrin complex and direct immunoprecipitation with antiserum to purified transferrin receptor from placentae. Sodium dodecyl sulfate polyacrylamide gel electrophoresis of the receptor isolated from reticulocytes reveals a polypeptide at Mr = 94,000 identical in molecular weight with that of the placenta. A radioimmunoassay using purified 125I-labeled transferrin receptor from placentae and antiserum to transferrin receptor fails to distinguish any immunological differences between the reticulocyte and placental forms of the protein. In addition, proteolytic digests of both of these polypeptides with Staphylococcus aureus protease show identical proteolytic patterns, indicating similar sequences. PMID- 6273398 TI - Unusual copper-induced sensitization of the biological damage due to superoxide radicals. PMID- 6273399 TI - Beta protein of bacteriophage lambda promotes renaturation of DNA. AB - The protein encoded by the red beta gene of bacteriophage lambda was found to promote reannealing of complementary single strands of DNA. Reannealing activity was optimal at pH 6.0 and required a divalent cation. A threshold temperature of at least 15 degrees C was necessary in order to detect activity. The reaction was linear with time for about 20 min, but the extent of reaction was dependent upon the amount of beta protein added. Reannealing of complementary single strands was confirmed by measuring increased lability of DNA to lambda exonuclease. When protein preparations from red- lysogens were tested for ability to promote reannealing, high activity was observed in a mutant altered in the gene for exonuclease, but there was low activity in a mutant altered in the gene for beta protein. PMID- 6273400 TI - cis-Polyunsaturated fatty acids induce high levels of superoxide production by human neutrophils. AB - Arachidonate stimulates the production of large quantities of superoxide by human neutrophils: 93.8 +/- 12.5 S.D. nmol of O2(-)/min/10(7) cells. This rate is comparable to that observed with the most effective neutrophil-stimulating agents previously reported. Other cis-unsaturated fatty acids are also capable of eliciting this response, the order of effectiveness being: arachidonate greater than gamma-linolenate greater than linoleate greater than oleate. Linolelaidate, myristate, and palmitate are ineffective. These data are discussed in relation to recent reports concerning the oxidation of arachidonic acid by human neutrophils and by a cell-free system that generates superoxide and hydrogen peroxide. PMID- 6273401 TI - Neutral cholesterol esterase activity in macrophages and its enhancement by cAMP dependent protein kinase. AB - Cholesterol esterase with optimal activity at pH 6.8 was found in the 40,000 X g supernatant fraction prepared from rabbit alveolar macrophages, thioglycolate elicited mouse peritoneal macrophages, and the J774 macrophage cell line. The neutral cholesterol esterase activity in these three types of macrophages was of the same order of magnitude as that in the 40,000 X g supernatant fraction of adrenal, heart, and liver but considerably lower than that in adipose tissue. The enzyme prepared from J774 cells was activated about 60% by preincubation with cAMP and Mg-ATP; half-maximal activation was obtained at 2.5 X 10(-7) M cAmP. The activation was completely blocked by a specific protein inhibitor of cAMP dependent protein kinase. These findings suggest that cytoplasmic cholesterol esters stored in macrophages can be mobilized for release by action of the neutral enzyme described and, further, that this process may be regulated by factors that affect intracellular cAMP levels. PMID- 6273402 TI - Cholinephosphotransferase in rat lung. The in vitro synthesis of dipalmitoylphosphatidylcholine from dipalmitoylglycerol. AB - Active substrate preparations of dipalmitoylglycerol were obtained by sonicating dipalmitoylglycerol, phosphatidylglycerol, and Tween 20 together at 65 degrees C. The apparent Vmax for cholinephosphotransferase in lung microsomes was 30 nmol/min/mg of protein for dipalmitoylglycerol-phosphatidylglycerol-Tween 20 substrate preparations and 43 nmol/min/mg of protein for dioleoylglycerol phosphatidylglycerol-Tween 20 preparations. Sonication at 65 degrees C was required for maximal activity with dipalmitoylglycerol but not for dioleoylglycerol. Highest activity was obtained when diacylglycerol was sonicated with phosphatidylglycerol and Tween 20 although phosphatidylglycerol also increased the activity when added separately. The presence of phosphatidylglycerol was critical particularly with dipalmitoylglycerol as substrate. Phosphatidylinositol, phosphatidylserine, and phosphatidic acid were slightly active but phosphatidylcholine and phosphatidylethanolamine were inactive. PMID- 6273403 TI - Purification and properties of protocatechuate 3,4-dioxygenase from Pseudomonas putida. A new iron to subunit stoichiometry. AB - Protocatechuate dioxygenase has been isolated from Pseudomonas putida. This new species of protocatechuate dioxygenase has been characterized and compared with the enzyme from Pseudomonas aeruginosa. The enzyme reported here has visible absorption, circular dichroism, electron paramagnetic resonance, and Raman spectroscopic properties virtually identical to those for protocatechuate dioxygenase from P. aeruginosa. However, the molecular weight and iron:subunit stoichiometry differ. Protocatechuate dioxygenase from P. putida has a molecular weight of 200,000 and contains 4 alpha subunits of 23,000 daltons, 4 beta subunits of 26,500 daltons, and 4 ferric irons suggesting that the enzyme is a tetramer of (alpha beta Fe+3) catalytic units. Protocatechuate dioxygenase from P. aeruginosa has been reported to have a Mr = 700,000, consisting of 16 alpha subunits of 22,500 daltons, 16 beta subunits of 25,000 daltons, and 8 ferric irons (Yoshida, R., Hori, K., Fujiwara, M., Saeki, Y., Kagamiyama, H., and Nozaki, W. (1976) Biochemistry 15, 4048-4053). This enzyme is thought to be an octamer of (alpha 2 beta 2 Fe+3) catalytic units, although reconstitution with extra iron will somewhat increase its activity. Using stopped flow techniques, we have shown that essentially all of the iron in the P. putida enzyme is catalytically active. This suggests that the minimal catalytic unit of all non heme iron intradiol dioxygenases is an (alpha beta Fe+3) structure. PMID- 6273404 TI - Copurification of alkaline phosphatase and 5'-AMP specific nucleotidase in Dictyostelium discoideum. PMID- 6273405 TI - Kinetic mechanism of assimilatory NADH:nitrate reductase from Chlorella. AB - Chlorella nitrate reductase catalyzes the reduction of nitrate to nitrite by NADH. Initial velocity studies showed that the kinetic mechanism is sequential, indicating that both substrates must bind to the enzyme before any products are released. Product inhibition with NAD and nitrite showed that competitive inhibition was observed when the inhibitor was similar to the varied substrate, while noncompetitive inhibition was observed when the inhibitor was dissimilar to the varied substrate. Likewise, dead-end inhibition with adenosine 5' diphosphoribose and thiocyanate showed competitive inhibition when the inhibitor was similar to the varied substrate and noncompetitive inhibition when the inhibitor was dissimilar to the varied substrate. These results indicate that Chlorella nitrate reductase follows a random bi bi kinetic mechanism. Phosphate was found to stimulate NADH:nitrate reductase activity and 2-fold. The NADH:cytochrome c reductase activity associated with nitrate reductase was not affected by phosphate suggesting the effect of phosphate is on the nitrate reducing moiety of the enzyme. Phosphate increases Vmax but has no effect on the apparent Km for nitrate. PMID- 6273406 TI - Characterization of a rat liver mitochondrial DNA-protein complex. Replicative intermediates are protected against branch migrational loss. AB - A stable DNA-protein complex was released from rat liver mitochondria by sodium dodecyl sulfate-lysis and isolated by sedimentation velocity in sucrose density gradients. The mtDNA-protein complex was washed with 0.5 M NaCl and any unbound contaminants were removed by hydroxyapatite column chromatography. The only detectable polypeptide in the complex was a single low molecular weight species (Mr = 16,000) having a slightly basic isoelectric point of 7.6-7.8. Complete digestion of the mtDNA-protein complex with restriction endonuclease HindIII revealed in agarose gels an "extra" band consisting of a subset of the largest fragment population. The fragments in this subset were shown to contain the replicative intermediates which were retarded in electrophoretic migration due to the parental strand separation in the region of the replication loops. No loss of nascent strands due to branch migration of the parental strands was observed upon HindIII cleavage of the covalently closed circular DNA in the mtDNA-protein complex. However, HindIII digestion of completely deproteinized mtDNA resulted in quantitative loss of nascent strands from replicating molecules. These results are interpreted as evidence that the single low molecular weight polypeptide present in the complex plays a major role in maintaining the integrity of replication loops during parental strand scission. PMID- 6273407 TI - Modulation of low density lipoprotein metabolism in bovine granulosa cells as a function of their steroidogenic activity. PMID- 6273409 TI - Structure of the Tetrahymena pyriformis rRNA gene. Nucleotide sequence of the transcription initiation region. AB - Portions of rRNA gene repeat from Tetrahymena pyriformis were cloned in pBR322, and a subclone containing the transcription initiation site was isolated. The promoter region was identified by S1 nuclease mapping of a portion of the HindIIIB fragment of the rRNA gene, using the 35 S rRNA primary transcript as a probe. The nucleotide sequence of the initiation site and the 5'-flanking region reveals three sets of highly conserved repeat sequences. A model of the promoter region is presented in which the rDNA is wrapped around a series of histone octamers. In this model the families of repeat sequences are brought in close proximity and suggest several possible mechanisms for the regulation of rRNA synthesis in T. pyriformis. PMID- 6273408 TI - Metabolic relationships between lipolysis and respiration in rat brown adipocytes. The role of long chain fatty acids as regulators of mitochondrial respiration and feedback inhibitors of lipolysis. AB - The calorigenic action of norepinephrine in isolated brown adipocytes was selectively mimicked by theophylline, dibutyryl cyclic AMP, and the principal fatty acids known to be present in the acyl moieties of brown adipose tissue triglycerides (palmitic, oleic, and linoleic acids). The stimulatory effects of fatty acids were entirely reversible, occurred at physiological concentrations, and were critically dependent upon the molar ratio of extracellular fatty acids to albumin. The calorigenic potency of fatty acids increased with their chain length. The apparent synchrony between the switching "on and off" of lipolysis and respiration by norepinephrine and propranolol indicated that the two phenomena are functionally inter-related and that they are both mediated by beta adrenoreceptors. Respiratory stimulation by palmitic acid was accompanied by an inhibition of glycerol release suggesting that fatty acids retroinhibit lipolysis while simultaneously activating respiration. Studies with 2-tetradecylglycidic acid, oligomycin, and uncouplers of oxidative phosphorylation support the view that fatty acids exert their calorigenic effects by increasing mitochondrial proton permeability and by simultaneously serving as substrates for beta oxidation via carnitine-dependent pathways. Since fatty acids mimicked the calorigenic action of norepinephrine even when beta-adrenoreceptors were blocked by propranolol, it is concluded that cyclic AMP controls respiration indirectly, most probably by modulating lipolysis. It is suggested that endogenous long chain fatty acids released in consequence of cyclic AMP activation of lipolysis play a fundamental role in the control of brown adipose tissue metabolism by self regulating lipolysis and by serving as physiological modulators of mitochondrial oxygen consumption. PMID- 6273410 TI - Structure of the Tetrahymena pyriformis rRNA gene. Nucleotide sequence of the transcription termination region. AB - The 3'-terminal regions of the 25 S rRNA and 35 S rRNA were mapped in the HindIIIC fragment of the T. pyriformis rRNA gene (Niles, E. G., and Jain, R. K. (1981) Biochemistry 20, 905-909). The precise locations of these termini were identified by S1 nuclease mapping. The 35 S rRNA was shown to contain two major 3'-ends, one identical to the 3'-end of the mature 25 S rRNA and one which is 15 bases longer. The nucleotide sequence of 440 base pairs of DNA in the terminator region was determined. There are several oligo T stretches and regions of dyad symmetry. Three models of transcription termination are presented which are based on possible secondary structures in the 3'-terminal region of the 35 S rRNA. PMID- 6273411 TI - Escherichia coli heat-labile enterotoxin. Ganglioside specificity and ADP ribosyltransferase activity. PMID- 6273412 TI - Gonadotropin-induced receptor regulation and steroidogenic lesions in cultured Leydig cells. Induction of specific protein synthesis by chorionic gonadotropin and estradiol. PMID- 6273413 TI - Biosynthesis of the human transferrin receptor in cultured cells. AB - The biosynthesis and degradation of the cell surface transferrin receptor has been investigated. The receptor is a glycoprotein, and evidence is presented that the mature receptor contains both complex and high mannose N-asparagine-linked oligosaccharides that are synthesized via a common high mannose intermediate as previously described for other glycoproteins. It is shown that fatty acid is associated with only the mature form of the receptor and that addition of fatty acid to the receptor and that addition of fatty acid to the receptor can occur as long as 48 h after synthesis. Glycosylation is not an absolute requirement for the receptor to act as acceptor for fatty acid, nor for transport to the cell surface, although the efficiency of both processes may be reduced in tunicamycin treated cells. The protein moiety of the transferrin receptor is degraded with a half-life of approximately 60 h. PMID- 6273415 TI - Sequence of the yeast iso-1-cytochrome c mRNA. AB - The nucleotide sequence of the yeast iso-1-cytochrome c (CYC1) mRNA is presented. The mRNA was enriched by hybridization to cloned CYC1 DNA attached to a solid matrix: either nitrocellulose filters or diazobenzyloxymethyl cellulose powder. The sequence of the 5'-end of the mRNA was determined by the extension of a CYC1 specific dodecanucleotide primer; the sequence of the 3'-end was determined using a decanucleotide d(pT8-G-A) primer. The CYC1 mRNA begins 61 nucleotides 5' to the AUG initiation codon, extends through the coding sequence to 172 to 175 nucleotides 3' to the UAA termination codon, followed by the poly(A) tail. There are no intervening sequences. Some of the sequences that the CYC1 mRNA shares in common with other eukaryotic mRNAs are discussed. PMID- 6273414 TI - The regulatory component of adenylate cyclase. Purification and properties of the turkey erythrocyte protein. AB - The regulatory component (G/F) of adenylate cyclase has been purified from turkey erythrocyte plasma membranes by adaptation of procedures developed for purification of the rabbit liver protein. The major modifications entail inclusion of high concentrations of NaCl to facilitate extraction and reconstitution of the protein. A typical preparation yields 200 micrograms of protein with a reconstitutive specific activity of 3-4 mumol . min-1 mg-1. Turkey erythrocyte G/F contains two putative subunits of 35,000 and 45,000 daltons. The 52,000-dalton polypeptide that appears to be a component of rabbit liver G/F is lacking. In solution, G/F behaves as a particle with Mr = 81,000. This value is reduced to 50,000 in the presence of activating ligands, suggesting dissociation of subunits. Activation of G/F by guanine nucleotide analogs is markedly accelerated in the presence of high concentrations of Mg2+. Reconstitutive and physical properties of the protein are also affected by fluoride. Cyc- S49 lymphoma membranes reconstituted with turkey erythrocyte G/F acquire properties that are characteristic of the turkey adenylate cyclase system; at least certain differing characteristics of adenylate cyclase systems are thus dictated by the nature of their G/F. PMID- 6273416 TI - Co-polymeric glycosaminoglycans in transformed cells. Transformation-dependent changes in the self-associating properties of cell-surface heparan sulfate. PMID- 6273417 TI - Vipoxin. A protein from Russell's viper venom with high affinity for biogenic amine receptors. AB - We have purified a small, basic protein with high affinity and selectivity for biogenic amine receptors to apparent homogeneity from the venom of Russell's viper (Vipera russelli). This protein, which we designate "vipoxin," has Mr = 13,000, and appears to exist in solution as a single polypeptide chain. It may contain 2 atypical amino acids. Vipoxin inhibits in a dose-dependent manner the binding of 3H-ligands to biogenic amine receptors, with apparent Ki values of 3 nM at alpha 1-adrenergic receptors, 5 nM at alpha 2-adrenergic receptors, 15 nM at dopamine receptors, and 32 nM at serotonin receptors. At concentrations up to 1 microM, vipoxin is inactive at beta-adrenergic, histamine, nicotinic cholinergic, muscarinic cholinergic, adenosine, gamma-aminobutyric acid, benzodiazepine, or opiate receptor binding sites. The effect of vipoxin is essentially irreversible over 20 h at alpha 1- and alpha 2-adrenergic receptors and serotonin receptors and is only slightly reversible at dopamine receptors. Norepinephrine protects alpha-adrenergic receptors from inhibition by vipoxin, while dopamine does not. Vipoxin has no protease activity but does have phospholipase A2 activity, which cannot account for its action on receptors, since receptor binding is assayed in the presence of 1 mM CoSO4 which completely and selectively inhibits the phospholipase activity. Other phospholipases A2 in the same venom lack vipoxin's action on receptors. In physiologic experiments, vipoxin behaves as an agonist at alpha 2-adrenergic receptors in the rat vas deferens and is over an order of magnitude more potent than norepinephrine itself. At alpha 1-adrenergic receptors, it is neither a simple agonist nor an antagonist, but selectively potentiates norepinephrine. Vipoxin may be a useful tool for biogenic amine receptor characterization. PMID- 6273418 TI - Translation of vesicular stomatitis and Sindbis virus mRNAs in cell-free extracts of Aedes albopictus cells. AB - We have developed a cell-free system from Aedes albopictus (mosquito) cells which is able to carry out endogenous protein synthesis and is stable to freezing and thawing. Successful preparation of extracts was found to depend on the presence of purified placental RNase inhibitor during cell breakage. Micrococcal nuclease treated extracts translated exogenously added Sindbis 26S or vesicular stomatitis virus mRNA with a high degree of fidelity, demonstrating that initiation of protein synthesis had occurred. Evidence is presented showing that when cell fractions containing intracellular membranes were used to translate vesicular stomatitis virus mRNA, the G protein was glycosylated and inserted into microsomal vesicles. Additional studies indicate that initiation of protein synthesis in this system is dependent on a capped and methylated 5'-terminal structure in the mRNA. PMID- 6273419 TI - Chain assembly intermediate in the biosynthesis of type III procollagen in chick embryo blood vessels. AB - A general mechanism for the assembly of procollagens is proposed from a biosynthetic study of procollagen III. This was shown to proceed by a stepwise process punctuated by disulfide bond formation and an assembly intermediate was recovered. The biosynthesis of type III procollagen in excised chick embryo blood vessels was studied by radioactive labeling for 30 min. Velocity sedimentation under denaturing conditions and purified antibodies specific against bovine amino propeptide III were used to identify and characterize monomeric pro alpha 1 III chains and a type III procollagen intermediate which is interchain disulfide linked only at the carboxyl end but not at the amino end. The monomeric chains presumably have intrachain disulfide bonds within the propeptides. The monomeric pro alpha 1 III chains were also found when alpha, alpha'-dipyridyl was present during incubation. Pulse-chase experiments show that the monomeric chains and the intermediate are biosynthetic precursors of type III procollagen. Furthermore, it is shown that monomeric pro alpha 1 chains are not triple helical when extracted under nondenaturing conditions. The results indicate that the assembly of pro alpha 1 III chains into type III procollagen starts with the association of the folded carboxyl propeptides and is followed by formation of disulfide bonds between carboxyl propeptides, folding of the triple helix, and formation of disulfide bonds between amino propeptides. All procollagens may follow a similar assembly sequence. PMID- 6273420 TI - DNA determinants important in sequence recognition by Eco RI endonuclease. AB - Alkylation interference and protection methods (Siebenlist, U., and Gilbert, W., (1980) Proc. Natl. Acad. Sci. U. S. A. 77, 122-126) have been utilized to deduce potential DNA contacts involved in specific complex formation between Eco RI endonuclease and its recognition sequence. The endonuclease protected the N7 position (major groove) of the dG and the N3 position (minor groove) of both dA residues within the Eco RI sequence against alkylation by dimethylsulfate, d(GpApApTpTpC), suggesting the presence of poly-peptide in both grooves in the vicinity of affected nitrogens. Results of methylation interference analysis suggest that the N7 of the Eco RI site dG and the N3 of the central dA, d(GpApApTpTpC), are utilized as contacts by the enzyme. The failure to observe interference upon methylation of the 5'-penultimate dA within the sequence implies that the endonuclease does not bond to the N3 of this residue, despite the fact that it is protected against alkylation by the protein. Ethylation interference patterns suggest four major phosphate contacts between endonuclease and each DNA strand. Two of these phosphates are 5'-external to the Eco RI sequence, d(pNpGpApApTpTpC), suggesting involvement of outside phosphates in electrostatic interactions. Moreover, alkylation protection and interference effects on the two DNA strands display perfect 2-fold symmetry. Thus, the endonuclease interacts with a minimum of 10 nucleotide pairs to yield a DNA protein complex characterized by elements of symmetry. In contrast, specific alkylation effects were not observed in comparable experiments with the endonuclease and a DNA which had been previously methylated by the Eco RI modification enzyme. PMID- 6273421 TI - Structural domains of vesicular stomatitis virus. A study by differential scanning calorimetry, thermal gel analysis, and thermal electron microscopy. AB - Differential scanning calorimetry has been used in combination with thermal gel analysis and electron microscopy to identify and study the structural domains of the membrane-enclosed vesicular stomatitis virus and its isolated internal components. Three major endothermic transitions centered at approximately 52, 76, and 80 degrees C and at least two minor transitions are observed at pH 7.0 for the intact virion. Thermal gel analysis suggests the possibility that specific proteins of vesicular stomatitis virus are involved in two or more of the calorimetric transitions. The effect of heating to defined temperatures on the morphology of the virion was studied by negative stain electron microscopy. The results of these "thermal EM" studies show discrete irreversible morphological changes in the virion which seem to coincide with the three major calorimetric transitions. PMID- 6273422 TI - Synthesis and assembly of membrane glycoproteins. Membrane anchoring COOH terminal domain of vesicular stomatitis virus envelope glycoprotein G contains fatty acids. AB - Two polypeptides associated with the envelope of vesicular stomatitis virus are obtained by exhaustive proteolytic digestion of the virion. Analysis of the tryptic peptides and determination of the partial amino acid sequence show that the larger membrane-anchoring peptide is derived from the hydrophobic COOH terminus of the viral transmembrane glycoprotein G. The smaller peptide is, however, derived from the nonglycosylated matrix protein M. Analysis of the membrane-anchoring peptide fragments obtained from virus labeled with [3H]palmitic acid shows that the larger peptide fragment contained all the fatty acid present in G, suggesting that the fatty acids in conjunction with the hydrophobic domain may be involved in the binding of G protein to the membrane. PMID- 6273423 TI - Isolation and characterization of proteolytically derived ileal receptor for intrinsic factor-cobalamin. PMID- 6273424 TI - alpha-Adrenergic activation of phosphorylase in liver cells involves mobilization of intracellular calcium without influx of extracellular calcium. PMID- 6273425 TI - Requirements for cholera toxin-dependent ADP-ribosylation of the purified regulatory component of adenylate cyclase. AB - The requirements for cholera toxin-catalyzed ADP ribosylation of the purified regulatory component of adenylate cyclase are described. In addition to the toxin, this reaction is dependent on or is facilitated by NAD, GTP, phospholipid, and a factor found associated with plasma membranes from several sources. Factor activity is heat-labile and protease-sensitive but is unaffected by treatment with N-ethylmaleimide. Gel filtration indicates that the factor behaves as a monodisperse species with a Stokes radius of 3.2 nm. The factor thus appears to be a protein that is distinct from any of the known components of adenylate cyclase. Factor activity was also detected in the cytoplasm of S49 cells. The cytoplasmic factor was smaller (Stokes radius = 2.0 nm) than the membrane-derived factor, and it was inactivated in the presence of sodium cholate. The initial rate of activation of the regulatory component of adenylate cyclase by toxin was found to be linearly related to the amount of factor present in the reaction. This has allowed the quantitation and partial purification (33-fold from detergent extracts) of the factor from turkey erythrocyte membranes. PMID- 6273426 TI - Influence of phosphorylation on the interaction of effectors with rat liver pyruvate kinase. PMID- 6273427 TI - Hydrolysis of esters by carboxypeptidase A requires a penta-coordinate metal ion. AB - The catalytic role of the metal ion in bovine carboxypeptidase A (peptidyl-L amino acid hydrolase; EC 3.4.12.2) is investigated by application of cryoenzymologic and electron paramagnetic resonance methods with use of the Co2+ reconstituted enzyme. Incorporation of 17O into oxygen-donor ligands induces a substantial change in the spin-lattice relaxation probability of the paramagnetic ion. While a change in spin-lattice relaxation is observed for the free Co2+ enzyme in 17O-enriched water, no change is observed for the enzyme complexed to glycyl-L-tyrosine. These results are consistent with x-ray crystallographic studies showing that the metal-bound water molecule in the active site is displaced upon binding of the peptide inhibitor. A change in spin-lattice relaxation of the Co2+ ion in the mixed anhydride, acyl-enzyme intermediate formed with the specific ester substrate O-(trans-p-chlorocinnamoyl)-L-beta phenyllactate is observed when 17O is enriched either into water or into the carbonyl oxygen position of the scissile bond of the substrate. Since the protein supplies three amino acid side chains as ligands to the metal ion, these results indicate that the metal ion is altered from a tetra-coordinate species in the free enzyme to a penta-coordinate species in the acyl-enzyme reaction intermediate. In addition, the results provide structural support for our assignment of ionization of a metal-bound water molecule in rate-limiting deacylation (Makinen, M. W., Kuo, L. C., Dymowski, J. J., and Jaffer, S. (1979) J. Biol. Chem. 254, 356-366) and affirm that the metal-hydroxide species is the nucleophile responsible for the breakdown of the mixed anhydride reaction intermediate of carboxypeptidase A. PMID- 6273428 TI - Epinephrine activation of phosphofructokinase in perfused rat heart. Properties of the activated and nonactivated forms and the interconverting reactions. PMID- 6273429 TI - The nucleotide sequences of rearranged and germline immunoglobulin VH genes of a mouse myeloma MC101 and evolution of VH genes in mouse. PMID- 6273430 TI - Differentiation-specific increase of cAMP-dependent protein kinase in the 3T3-L1 cells. PMID- 6273431 TI - Receptor-independent catabolism of low density lipoprotein. Involvement of the reticuloendothelial system. AB - In this study we examine the effects of intravenous ethyl oleate emulsions on the metabolism of native and cyclohexanedione-modified human low density lipoprotein in rabbits. Treatment produced a highly significant fall in receptor-independent catabolism as measured by the fractional clearance rate of cyclohexanedione modified low density lipoprotein. Receptor-dependent catabolism (the difference between the fractional clearance rates of native and cyclohexanedione-modified low density lipoprotein) was variably affected with some animals showing a decrease in receptor activity. These data suggest that the reticuloendothelial system makes a substantial contribution to receptor-independent low density lipoprotein catabolism in the rabbit. PMID- 6273432 TI - Identification of lysine residues in the binding domain of ribonuclease A for the RNase inhibitor from human placenta. AB - Amidination of the available lysine residues of the complex between RNase A and human placental RNase inhibitor has been performed with methyl acetimidate; the conditions of the derivatization preserve the complex functionally intact. Resistance of epsilon-acetimidyllysine residues to hydrolysis by trypsin allowed, after peptide mapping, the identification of lysine residues 7, 31, 41, 61, and 91 as those which were fully protected by the inhibitor from amidination. Lysine residue 37 was partially protected from amidination. In the presence of poly(A), lysine residues 41 and 61 of RNase A were fully protected from amidination, while lysine residues 7, 31, 37, 91, and 104 were only partially protected; the enzyme retained full activity. The results permit identification of lysine residues located in the binding domain of RNase A for the inhibitor. This region is not identical with, but does overlap, the binding domain for poly(A). PMID- 6273433 TI - Choleragen-stimulated release of guanyl nucleotides from turkey erythrocyte membranes. AB - Choleragen stimulates adenylate cyclase by ADP ribosylating a guanyl nucleotide binding regulatory protein (G/F). beta-Adrenergic hormones also activate the adenylate cyclase of turkey erythrocytes, and it is currently believed that they do so in part by decreasing the affinity of G/F factor for GDP, an effect which is manifested by a hormone-stimulated release of guanyl nucleotides from the membranes. Since choleragen might also activate adenylate cyclase by a similar mechanism, the effect of toxin treatment on the release of guanyl nucleotides from turkey erythrocyte membranes was examined. In the presence of NAD, choleragen was found to stimulate release of guanyl nucleotides from membranes which had been preloaded with radiolabeled GTP. No stimulation of release was observed with cAMP or when NAD was replaced by NADP, which does not serve as a substrate for choleragen-catalyzed ADP ribosylation. While either isoproterenol or choleragen can stimulate release of guanyl nucleotides from the membranes, the amount of guanyl nucleotide released in the presence of both isoproterenol and choleragen was no greater than that released by isoproterenol alone. Furthermore, when membranes were first treated with choleragen and NAD, the subsequent release of guanyl nucleotides induced by isoproterenol was reduced to approximately 15% of that observed with membranes not treated with the toxin. Therefore, choleragen may enhance release of guanine nucleotides from sites on the membranes that are also affected by beta-adrenergic agonists, sites which are thought to correspond to G/F. These data are consistent with the hypothesis that choleragen may stimulate adenylate cyclase, in part, by enhancing release of guanyl nucleotides, a mechanism similar to that of beta-adrenergic agonists. PMID- 6273434 TI - Protein kinase activity and substrates at the surface of intact HeLa cells. AB - Evidence is presented for the location at the surface of HeLa cells of a protein kinase capable of phosphorylating surface as well as extracellular (foreign) proteins. The reaction products have been found to be proteins containing phosphoryl groups as monoesters of seryl and threonyl residues (but not of tyrosine). The enzyme is of the cyclic AMP-independent type, since neither cyclic AMP nor the heat- and acid-stable inhibitor protein (specific for cyclic AMP dependent protein kinases) influenced its activity. Further, co-substrate ATP could in part be substituted by GTP, and the spectrum of proteins phosphorylated by the ecto-enzyme differed from that phosphorylated by cyclic AMP-dependent protein kinases. Evidence for the ecto-enzymic nature of this protein kinase includes (a) utilization of co-substrate and location of products at the surface of cells carefully controlled as being in an intact state and (b) phosphorylation of exogenous protein (phosvitin; specific serum proteins) by intact cells. Conclusive proof was gained by qualitative and quantitative comparative studies of phosphorylation in cultures with varying degrees of damaged cells either as a whole or after separation into groups of intact and damaged cells by electronic cell sorting. The results of experiments with cell sonicates excluded the possibility that either enzyme or substrates released from damaged cells were simply adsorbing to the cell surface. PMID- 6273435 TI - A study of the electron paramagnetic resonance properties of single monoclinic crystals of bovine superoxide dismutase. AB - Monoclinic crystals of native bovine superoxide dismutase and its monocyano derivative were studied by means of electron paramagnetic resonance spectroscopy. Through computer simulation of the spectra, the directions of the principal axes of the magnetic tensors (g and A) have been found with respect to the crystal principal axes and with respect to the positions of atoms bear the Cu(II) as previously determined by x-ray crystallography (Richardson, J. S., Thomas, K. A., and Richardson, D. C. (1975) Biochem. Biophys. Res. Commun. 63, 986-992; Tainer, J. A., Getzoff, E. D., Richardson, J. S., and Richardson, D. C. (1980) in 2SOD: Cu, Zn-Superoxide Dismutase Complete Atomic Coordinates (Richardson, D. C., and Richardson, J. S., eds) Brookhaven Protein Structure Data Bank). In the native protein, the direction of the gz axis of Cu(II) was found to lie perpendicular to the rough plane formed by the four imidazole nitrogen atoms coordinated to the Cu(II). The direction of gy is approximately along the His 44N-Cu-His 46N direction, and gx is in the direction of the Cu-His 61-Cu-N bond. The A is coaxial with g within 15 degrees C. A substantial shift occurs in the direction of gz when CN- binds to the Cu(II), suggesting a change in the coordination configuration of the metal. PMID- 6273436 TI - Insulin decreases phosphoenolpyruvate carboxykinase (GTP) mRNA activity by a receptor-mediated process. AB - The mRNA that codes for phosphoenolpyruvate carboxykinase accounts for approximately 0.2% of the protein synthesized in H4IIEC3 hepatoma cells maintained for 24 h in serum-free medium containing N6,O2'-dibutyryl cAMP and theophylline. This value decreases to 0.04% within 3 h after the addition of insulin. Maximal effects are produced by 10(-10) M insulin, and half-maximal deinduction of both the relative rate of synthesis of P-enolpyruvate carboxykinase and mRNA coding for P-enolpyruvate carboxykinase activity occurs at approximately 2 X 10(-12) M insulin. Porcine proinsulin is 4% as potent as porcine insulin since half-maximal deinduction of mRNA coding for P-enolpyruvate carboxykinase occurs at 5 X 10(-11) M. The concentration of proinsulin required to inhibit 125I-insulin binding by 50% is 2 X 10(-7) M, as compared to 6 X 10(-9) M for insulin; thus, the decreased sensitivity of this deinduction to proinsulin parallels the decreased binding affinity H4IIEC3 cells have for proinsulin as compared to insulin. These data indicate that insulin regulates P-enolpyruvate carboxykinase synthesis through a receptor-mediated process, that the effect occurs when less than 2% of the insulin receptors are occupied, and that this effect is exerted prior to the level of mRNA translation. PMID- 6273437 TI - Structure of the tyrosyl radical in bacteriophage T4-induced ribonucleotide reductase. AB - Ribonucleotide reductase induced by bacteriophage T4 in Escherichia coli contains an organic free radical necessary for enzymatic activity. Its EPR spectrum at 77K is similar to but not identical with that of the corresponding radical in the enzyme from uninfected E. coli studied previously. Isotope substitutions now show that the radical in the T4-induced enzyme also is localized to a tyrosine residue with its spin density delocalized over the aromatic ring of tyrosine. The difference between the radicals of the T4-induced and the E. coli ribonucleotide reductases, as reflected in the hyperfine patterns of their EPR spectra, is suggested to be due to slightly different radical geometries, resulting from a twist of about 10 degrees around the bond between the aromatic ring and the methylene group in the tyrosine radical. Hydroxyurea destroys the free radicals of both ribonucleotide reductases and also their catalytic activities. Both enzymes are considerably more sensitive to hydroxyurea during catalysis than in the noncatalytic state. However, when compared to the bacterial ribonucleotide reductase, the T4-induced enzyme shows an overall approximately 10 times higher sensitivity to hydroxyurea, judging from the drug concentrations needed to destroy the radicals and inhibit the activities. This result may reflect a difference in accessibility for the drug to the active sites of the enzymes. PMID- 6273438 TI - Massive accumulation of phosphatidic acid in conditionally lethal CDP-diglyceride synthetase mutants and cytidine auxotrophs of Escherichia coli. AB - Escherichia coli mutants partially defective in CTP: phosphatidic acid cytidylyltransferase (CDP-diglyceride synthetase) are more resistant to the antibiotic erythromycin than are isogenic wild type strains. When 100 micrograms/ml erythromycin is added to nutrient agar plates, it is possible to obtain a 30-fold enrichment for cds mutants from a mutagen-treated stock, as judged by colony autoradiography (Ganong, B. R., Leonard, J. M., and Raetz, C. R. H. (1980) J. Biol. Chem. 255, 1623-1629). Using this approach, we have isolated 38 new cds mutants, nine of which are unable to grow at a culture pH greater than 8. A typical conditionally lethal mutant like GN80 contains a 3 to 5% phosphatidic acid below pH 7. Above pH 8, GN80 accumulates phosphatidic acid to about 30% of the total membrane lipid, while the de novo syntheses of phosphatidylethanolamine and phosphatidylglycerol are abruptly inhibited by over 10-fold. GN80 loses viability after 60 min at pH 8.5, and the liponucleotide pool of GN80 is about one-seventh that of an isogenic wild type, GN85, under these conditions. The pH optimum of the residual CDP-diglyceride synthetase present in extracts of GN80 is 0.5 pH units lower than normal. Twenty-one of 26 spontaneous pH-resistant revertants of GN80 concomitantly regain parental levels of the enzyme. Our results constitute definitive physiological proof that CDP diglyceride is an obligatory precursor for over 90% of the phosphatidylethanolamine and phosphatidylglycerol in E. coli. Independent evidence for this is provided by the observation that cytidine auxotrophs, which are defective in the conversion of UTP to CTP, also accumulate very high levels of phosphatidic acid after 1 h of cytidine starvation. PMID- 6273439 TI - Heart cytochrome c oxidase. An infrared study of effects of oxidation state on carbon monoxide binding. PMID- 6273440 TI - Direct linkage of 125I-EGF to cell surface receptors. A useful artifact of chloramine-T treatment. PMID- 6273441 TI - Rat glutathione S-transferase. Cloning of double-stranded cDNA and induction of its mRNA. AB - Messenger RNA extracted from the livers of normal, phenobarbital-treated, and trans-stilbene oxide-treated rats was translated in a mRNA-dependent protein synthesizing system. Immunoprecipitation of the translation products by antibodies against the Ya and Yc subunits of glutathione S-transferase detected two polypeptides of molecular weights 23,500 and 25,000. Subsequently, a clone containing glutathione S-transferase sequences was identified from a rat liver double-stranded cDNA library that had been prepared by homopolymeric tailing and cloning into the Pst I site of pBR322. Confirmation of the identity of the clone was obtained by recloning the 550-bp insert DNA into the phage vector M13 and utilizing the single strand recombinant phage DNA in specific hybrid selection of mRNA followed by translation and immunoprecipitation with antibodies to the Ya and Yc subunits. This recombinant phage, M13GST94, was also utilized in a new technique to synthesize 32P-labeled cDNA specific to the glutathione S transferase insert DNA that was used subsequently in RNA excess solution hybridization to determine the relative concentration of glutathione S transferase mRNA. Phenobarbital treatment resulted in a 3.2-fold increase in glutathione S-transferase mRNA over levels found in control rats, while trans stilbene oxide increased glutathione S-transferase mRNA levels 5.7-fold. The DNA sequence of the clone was determined and utilized to propose a partial amino acid sequence. PMID- 6273442 TI - Isolation of multiple genomic sequences coding for chicken myosin heavy chain protein. AB - A genomic bank has been constructed using DNA isolated from a dystrophic strain of chickens. The library was screened for myosin heavy chain (MHC) sequences using a cDNA probe and 11 positive recombinants isolated. Identity of the clones was confirmed by positive RNA selection via hybridization of the clones with muscle RNA and subsequent translation of the hybridizable RNA in vitro. Restriction maps indicate that the clones can be divided into 7 distinct groups; some of these groups do, however, share similar or homologous sequences. Hydridization of the clones to RNA derived from leg, breast, heart, and brain reveals that all of the clones code for the muscle-specific MHC isoforms. These experiments also show that there are at least 3 size classes of MHC mRNA sequences, whose relative amounts appear to be modulated in a tissue- and developmental stage-specific manner. PMID- 6273443 TI - Characterization of the action of Escherichia coli DNA polymerase I at incisions produced by repair endodeoxyribonucleases. PMID- 6273444 TI - Generation of superoxide via the interaction of nitrofurantoin with oxyhemoglobin. AB - Nitrofurantoin was found to interact with HbO2 to cause the concomitant formation of methemoglobin and superoxide. The rate of formation of methemoglobin and superoxide was linearly dependent upon the concentration of nitrofurantoin and could be inhibited by superoxide dismutase, catalase, or the prior conversion of HbO2 to ethylioscyanoferrohemoglobin. The ability of nitrofurantoin to interact with HbO2 and cause superoxide formation may represent one mechanism by which it produces red cell toxicity and suggests that generation of superoxide in erythrocytes may occur via a different mechanism than that which occurs in microsomes. PMID- 6273445 TI - The in-vitro degradation of poly(glycolic acid) sutures--effect of pH. AB - Tensile strength of poly(glycolic acid) suture (PGA) of size 2-0 was examined as a function of three pH levels, 5.25, 7.44, and 10.09 of the buffer. Cord and yarn grip was used to eliminate grip-induced failure of breaking strength tests. It was found that Dexon sutures degraded significantly faster in pH = 10.09 buffer than the other two lower pH buffers. There was no significant difference in degradation rate at pH = 5.25 and 7.44. At 7 days, PGA sutures lost almost half of its original tensile strength at pH = 10.09, while the same sutures still remained more than 95% of their original breaking strength at buffers of pH = 5.25 and 7.44. After 21 days, no trace of sutures could be detected in the buffer of pH = 10.09 while about 20% strength still remained in the buffers of pH = 7.44 and 5.25. Cage effect in the crystalline phase and pH dependent hydrogen bonding were introduced to explain the difference in degradation phenomenon of PGA at buffers of various pH. PMID- 6273446 TI - Benign fibrous histiocytoma of bone. Light- and electron-microscopic observations. AB - Case report of a patient with an unusual, rapidly growing bone tumor in the third and fourth cervical vertebrae. Histological and electron-microscopic investigations reveal a tumor composed of histiocytic cells, xanthomatous cells, giant cells of Touton type, and fibroblastic cells. No cellular features of malignancy are observed. From its cytologic appearance the tumor has to be classified as a benign fibrous histiocytoma. Regarding its ultrastructural features, the tumor may not be distinguished from non-ossifying fibroma of bone, but its clinical pattern shows obvious differences of localization and growth potential. the term "benign fibrous histiocytoma of bone" is proposed for these tumors which must be differentiated from non-ossifying fibroma. PMID- 6273447 TI - Rapid and reversible reduction of junctional permeability in cells infected with a temperature-sensitive mutant of avian sarcoma virus. AB - The transformed or normal phenotype of cultured normal rat kidney cells infected with a temperature-sensitive mutant of avian sarcoma virus is conditional on the temperature at which the cells are grown. Using dye injection techniques, we show that junction-mediated dye transfer is also temperature-sensitive. The extent and rate of transfer between infected cells grown at the transformation-permissive temperature (35 degrees C) is significantly reduced when compared to infected cells grown at the nonpermissive temperature (40.5 degrees C) or uninfected cells grown at either temperature. Infected cells subjected to reciprocal temperature shifts express rapid and reversible alterations of dye transfer capacities, with responses evident by 15 min and completed by 60 min for temperature shifts in either direction. These results suggest that altered junctional capacities may be fundamental to the expression of the ASV-induced, transformed phenotype. PMID- 6273448 TI - Quantitative cell fusion: the fusion sensitivity (FS) potential. AB - The dose response of Sendai virus-induced cell fusion was studied in 10 mammalian cell lines, comprising 5 continuous and 5 diploid cell lines originating from 5 species. The extent of fusion was calculated using a parameter directly proportional to the number of fusion events (t-parameter). At lower levels of fusion the dose response was found to be based on the same simple kinetic rules in all cell lines and was defined by the formula: t = FS . FAU/(I + FS . FAU), where FS (fusion sensitivity) is a cell-specific constant of the fusion rate and FAU (fusion activity units) is the virus dose. The FS potential of a cell line was determined as the linear regression coefficient of the fusion index (t/(I - t)) on the virus dose. At higher levels of fusion, when the fusion extent reached cell-line-specific maximal levels, the dose response was not as uniform. In general, and particularly in the cases of the diploid cell lines, these maximal levels were directly proportional to the FS potentials. Thus, it was concluded that the FS potential is the basic quantitative feature, which expresses the cellular fusion efficiency. The fact that FS varied extensively between cell lines, but at the same time apparently followed certain patterns (being higher in continuous compared to diploid cell lines and being related to the species of origin of the cells), emphasizes it biological significance as well as its possible usefulness in studies of the efficiency of various molecular interactions in the cell membrane/cytoskeleton system. PMID- 6273449 TI - Varicella-zoster-associated encephalitis: detection of specific antibody in cerebrospinal fluid. AB - Varicella-zoster (VZ) antibody measured by indirect immunofluorescence was used to identify patients with encephalitis due to VZ virus. In 15 of 16 (94%) patients with VZ infection and clinical evidence of central nervous system involvement, VZ antibody was present at a titer of 1:2 or greater in cerebrospinal fluid. In contrast, no VZ antibody was detected in the cerebrospinal fluid of seven patients with VZ infections without clinical signs of central nervous system involvement, nor was any detected in the cerebrospinal fluid of 25 patients with malignant or demyelinating disease or encephalitis due to other viral or toxic agents. All of these patients had detectable serum VZ antibody. Thus, in this study the presence of detectable VZ antibody in cerebrospinal fluid measured by indirect immunofluorescence seemed to correlate with clinical evidence of central nervous system involvement due to this virus. PMID- 6273450 TI - Comparison of techniques for recovering murine cytomegalovirus from a macrophage enriched subpopulation of mice. AB - We previously demonstrated the transmission of murine cytomegalovirus to syngeneic mice and preformed monolayers of mouse embryo fibroblasts with plastic adherent peritoneal exudate cells from infected mice as a source of macrophages. In the present studies we compared this standard feeder layer method with a reverse feeder layer method in which the adhering peritoneal exudate cells remained attached to plastic and to which were added mouse embryo fibroblasts. Recovery of virus from the adherent peritoneal exudate cells of infected mice was achieved with both methods. Whereas the standard method achieved greater accuracy and usually recovered more virus, the reverse method appeared to recover virus more frequently and required fewer cells to perform the assay. Furthermore, the reverse method demonstrated the survival of murine cytomegalovirus in adhering cells after culture periods of up to 2 weeks. PMID- 6273451 TI - Clostridium difficile toxin as a confounding factor in enterovirus isolation. AB - A peculiar cytotoxic effect, occasionally encountered in the course of inoculating cell cultures with fecal specimens for routine enterovirus isolation attempts, was shown to be produced by Clostridium difficile toxin. PMID- 6273452 TI - Bacteriology of acute obstructive suppurative cholangitis of the aged. AB - Bacteriological examination was performed on bile from 23 patients with acute obstructive suppurative cholangitis. Of 23 bile cultures, 15 yielded aerobic and anaerobic bacteria and 8 yielded aerobic bacteria only. No specimen grew anaerobic bacteria only. A total of 20 cultures yielded a polymicrobial flora, and 3 cultures grew one single aerobic bacterium. Escherichia coli, Klebsiella sp., enterococci, Bacteroids fragilis, and Clostridium perfringens were the predominant bacterial flora in bile of these patients. Bacteremia was documented in 10 of the 12 patients cultured. All bacteremias involved aerobic bacteria and were polymicrobial in three patients. The frequent presence of anaerobes in bile of patients with acute obstructive suppurative cholangitis suggests that antimicrobial therapy should provide adequate coverage for anaerobic bacteria and enteric organisms. PMID- 6273453 TI - Comparison of anticomplement immunofluorescence and fluorescent antibody-to membrane antigen tests for determination of immunity status to varicella-zoster virus and for serodifferentiation of varicella-zoster and herpes simplex virus infections. AB - The anticomplement immunofluorescence (ACIF) test was compared with the fluorescent antibody-to-membrane antigen (FAMA) test for determining varicella zoster virus antibody levels as a measure of varicella-zoster virus immunity status. The ACIF test was found to be comparable to the FAMA test in sensitivity and could be used for examining sera at low dilutions of 1:2 and 1:4. In addition, the ACIF method proved to be a more economical procedure in terms of antigen required and personnel time necessary to perform the test. Heterologous varicella-zoster virus antibody titer rises were demonstrated by the FAMA test with 10 serum pairs from patients with clinically diagnosed genital herpes simplex virus infection, indicating that the FAMA test is no more suitable than other serological methods for serodifferentiation of those herpes simplex virus and varicella-zoster virus infections in which antibody increases occur to both antigens. PMID- 6273454 TI - Determination of immunoglobulin M antibodies against hepatitis B core antigen and hepatitis A virus by reorienting sucrose gradient high-speed centrifugation for diagnosis of acute viral-hepatitis. AB - Immunoglobulin M (IgM) antibodies against hepatitis B core antigen (anti-HBc) and hepatitis A virus (anti-HAV) were determined in 41 cases of acute viral hepatitis. In sera positive for anti-HBc or anti-HAV, IgM was separated from IgG by reorienting sucrose gradient high-speed centrifugation, and the IgG- and IgM containing serum fractions were tested for the presence of specific antibody by radioimmunoassay. At the onset of illness, 4 of the 41 cases were classified as hepatitis A, 31 were hepatitis B, and 6 were non-A, non-B hepatitis, based on the results of these tests and of assays for hepatitis B surface antigen and antibody and hepatitis B e antigen and antibody. Fourteen of these 41 patients (34%) required IgM anti-HBc or IgM anti-HAV testing or both for appropriate classification. IgM anti-HBc persisted for at least 7 weeks after onset but no longer than 17 weeks in all patients tested with transient hepatitis B surface antigen-positive acute hepatitis. IgM anti-HAV persisted up to but not longer than 62 days in the patients with hepatitis A. Therefore, IgM anti-HBc and IgM anti-HAV determinants are valuable tools for the differential diagnosis of acute A, B, and non-A, non-B hepatitis. PMID- 6273455 TI - Autoantibody to nerve tissue in a patient with a peripheral neuropathy and an IgG paraprotein. AB - The antibody activity of a benign IgG lambda paraprotein to nerve tissue in a case of peripheral neuropathy has been investigated using immunohistochemical methods on tyrpsin-treated, formalin-fixed, paraffin-embedded tissue. IgG lambda was found in th sural nerve biopsy of the patient. Specific binding of the purified IgG lambda paraprotein and its isolated F(ab')2 fragment to homologous nerve and brain tissue was demonstrated. Similar activity was not demonstrable on fresh frozen cryostat sections. The results suggest that tests for autoantibodies to nerve tissue in neuropathological disorders should not be confined to fresh frozen tissue substrates. PMID- 6273456 TI - Phase I study of ceftizoxime, a new cephalosporin. Single-dose study. AB - A phase I study of ceftizoxime, a new cephalosporin, was performed in 29 subjects. No abnormalities were observed in subjective symptoms, laboratory test results, or physical test results at estimated therapeutic doses of 500 mg intramuscularly, 500 and 1000 mg intravenously, and 2000 mg drip infusion. It is concluded, therefore, that the drug is safe for clinical use. The mean peak serum concentration was dose dependent. The mean serum concentrations of ceftizoxime in man at 5 minutes after 500 and 1000 mg by intravenous bolus were 58.4 and 112.8 micrograms/ml, respectively, which exceed the MIC80 against most pathogens tested. Thus, 500 or 1000 mg was estimated to be the therapeutic dose. The distribution volume of ceftizoxime was 16.0 to 18.6 liters, the total clearance was 135.6 to 154.9 ml/min, and the half-life in the beta-phase was 1.36 to 1.39 hours. Ceftizoxime is mainly excreted in the urine as unchanged drug, at an excretion rate in the 24-hour urine of approximately 80 per cent. PMID- 6273457 TI - Ipsilateral corticocortical projections related to binaural columns in cat primary auditory cortex. AB - Projections from the high-frequency representations of auditory cortical fields A and P were autoradiographically labeled using anterograde transport of a mixture of tritiated proline and leucine. A single injection of isotope into either cortical field resulted in multiple regions of dense labeling within ipsilateral AI. As the result of an injection of radioactive label into field A in one experiment, densely labeled regions were found to be systematically related to the binaural map throughout the high-frequency representation of AI. Contralateral dominant suppression responses were located in regions of dense labeling while summation responses were located in regions of less dense labeling. In contrast, it was previously found that callosal axon terminations were more densely concentrated in summation columns than in contralateral dominant suppression columns (Imig and Brugge, '78). Thus, the two classes of binaural columns differ with respect to the density of innervation they receive from these ipsilateral and contralateral populations of neurons. In other experiments in which isotope was injected into field A, a systematic relationship between density of ipsilateral labeling and binaural response class was only seen in a portion of AI; in other regions no relationship was evident. A simple interpretation of these data follows. Within a contiguous territory in field A is a population of neurons whose axons provide more dense innervation to contralateral dominant suppression columns than to summation columns in ipsilateral AI. Injections of radioactivity confined to this territory result in a systematic relationship between the density of labeling and the binaural map throughout AI. Outside this territory is a population of neurons whose axon terminations are not systematically related to the binaural map in AI. Isotope injections which engage both territories may result in a systematic relation between density of labeling and the binaural map in one portion of AI, while in another, no relationship may be evident. There is some indication that projections from field P may also be related to binaural columns in AI in the same manner as are the projections from field A. PMID- 6273458 TI - Superior colliculus neurons which project to the cat lateral posterior nucleus have varying morphologies. AB - We have studied the laminar position, morphology, and synaptic relationships of neurons in the cat superior colliculus which project to the interadjacent division of the lateral posterior nucleus (LPi), using the retrograde transport of horseradish peroxidase. The neurons which project to LPi are remarkably varied in depth, size, morphology, and synaptic density and appear to consist of at least four cell types. Labeled cells were found laminae. Forty-seven percent were found in the superficial gray layer (50-550 micrometer), all but a few within its deep subdivision. Forty-seven percent were located in the optic layer (550-1,200 micrometer), the majority of these being within the upper one-half of the layer Seven percent were found in the intermediate and deep gray layers (below 1,200 micrometer). Cell body area varied widely, ranging from 37 to 768 micrometer 2 (mean of 243 micrometer2). Based on cell size, shape, and dendritic field orientation, we identified four distinct cell morphologies which were labeled. Thirty-five percent were stellate, 32% were vertical fusiform 19% were granule, and 12% were horizontal cells. Electron microscope analysis confirmed that neurons projecting to the lateral posterior nucleus are a morphologically diverse group. A sample of 71 labeled cells varied significantly in density of synaptic input as well as in size, shape, depth, dendritic distribution, and cytology. PMID- 6273459 TI - Thalamic projections to the primary and secondary somatosensory cortices in cat: single and double retrograde tracer studies. PMID- 6273460 TI - Annular hemosiderotic histiocytoma. AB - A 35-year-old woman was examined because of a symptomless multiple annular eruption. The lesion consisted of rings of closely set, smooth, firm papules that were brown in color. Hemosiderotic histiocytoma was diagnosed by light and electron microscopy. Ultrastructurally, the lesion consisted of fibroblastic and histiocytic cells with marked hemosiderin and lipid deposition. PMID- 6273461 TI - Fine structure of granular cell tumor of Abrikosov. AB - The case report illustrates the microscopical and ultrastructural findings obtained in a case of Abrikossoff's tumor. On the basis of the light and ultrastructural data a neural histogenesis is suggested. PMID- 6273462 TI - Melanocytic involvement in giant axonal neuropathy. AB - The skin of a patient with giant axonal neuropathy was studied by light and electron microscopy. Enlarged melanocytes protruded into the papillary dermis. An increased number of microfilaments (100 A), which accumulated in rounded or irregular clusters was the main pathological feature. Melanosomes and melanisation were normal. A similar increase of microfilaments was seen in axons and Schwann cells of peripheral nerves, fibroblasts and endothelial cells. This suggests that this metabolic, probably inborn and genetic disorder, does not only affect the nervous system but also the whole microfilament system (cytoskeleton). PMID- 6273463 TI - Scanning electron microscopy of acid detergent fiber digestion by rumen microorganisms. PMID- 6273464 TI - Isolation of peroxisomes from rat liver using sucrose and Percoll gradients. AB - Peroxisomes were isolated from the livers of both control and clofibrate-treated rats. Two procedures, one with a sucrose gradient, and a second with Percoll gradients, were utilized. The Percoll procedure allowed contamination of the isolated peroxisome fraction on protein basis, by lysosomes (8%), by mitochondria (5%) and by microsomes (2%). The peroxisome fraction isolated by the sucrose gradient showed no significant contamination with mitochondria, but the fraction contained 13% microsomes. In addition to established peroxisomal enzymes, the isolated peroxisomes also contained cytochrome b5, NADH-cytochrome c reductase and NADPH-isocitrate dehydrogenase. The peroxisomal membranes were also separated from the content, and they were found to have a relatively high phospholipid/protein ratio (0.55). PMID- 6273465 TI - Nervous regulation of insulin release by the intestinal vagal glucoreceptors. AB - In anesthetized cats and rats, it is demonstrated that glucose perfusion of the small intestine produces a rapid increase of insulin secretion (IRI) which precedes glycemia variation. This mechanism involves the autonomic nervous system and originates from intestinal glucoreceptors, the existence of which was recently reported. The nervous pathways are described in this study:(1) the afferent pathway is represented by vagal fibers coming from the intestinal glucoreceptors; (2) the efferent pathway involves both sympathetic fibers (splanchnic nerves) and chiefly parasympathetic fibers (vagal nerves). These results are established after surgical suppression of afferent and efferent vagal fibers, and pharmacological exclusion of parasympathetic or sympathetic fibers. The role of this nervous regulation of insulin secretion is discussed with special reference to other already known mechanisms. PMID- 6273466 TI - [Anything new concerning the human lens and senile cataract (author's transl)]. AB - Many differences exist between human and animal lenses. Firstly, ATP concentrations in human lenses remain relatively unchanged during aging, despite slowing down of carbohydrate metabolism. Secondly, growth rate, expressed either by fresh weight or by volume, decreases with age, but this evolution is much less pronounced in human than in animal lenses and in the former never stops completely. This protein synthesis appears to be reduction in a very important pathogenic factor in senile cataract. This opinion is strengthened by the fact that ATP, which hardly decreases with age in human lenses, diminishes only in the last stages of senile cataract, whereas the deficiency in protein synthesis is an early phenomenon, and precedes the first opacities. Incorporation of amino acids into soluble lens protein in vitro can be stimulated by adding to the culture medium, 5 m M dibutyryl c AMP protected by 0,5 m M isobutylmethylxanthine. Topical application to the eye of a few drops confirmed penetration of the c AMP into the lens. The theoretical possibility, always denied, of a preventive or stabilizing medicinal treatment for senile cataract, does, therefore, exist. PMID- 6273467 TI - Familial dermatoglyphic analysis in syndactyly type I. AB - A familial dermatoglyphic study of 25 syndactyly type I patients and 53 unaffected first-degree relatives showed a significant increase of complex fingertip patterns in the patients and in their relatives as well. An embryonic link between the formation of complex fingertip patterns and syndactyly type I is postulated. The study contributes new data on the hereditary transmission of syndactyly type I which cannot be used for individual genetic counseling. Calculation of the recurrence risk is difficult in view of the reduced penetrance of the gene. PMID- 6273468 TI - [The effect of conditioning on the ultrastructural distribution of synaptic vesicles in the hippocampus of the rat]. AB - In three months old male Wistar-rats the influence of conditioning (brightness discrimination reaction) on the number and distribution of vesicles of the synapses in the stratum radiatum (CA 3) of the hippocampus were investigated. A total of 15 rats (5 trained rats, 5 passive and 5 active controls) were studied by the aid of electron microscopic and morphometric methods. The number of synaptic vesicles was evaluated for the passive controls by 27 vesicles per synaptic area. In the CA 3-region 70 minutes after training the calculation of the number of synaptic vesicles revealed a significant decrease in comparison with passive and active controls (16 and 32 vesicles per synaptic area, respectively; significant difference at p less than 0.05). The volume of the synaptic vesicles was reduced for the trained rats by 20-25% in comparison with controls. The morphological results after training could be correlated with the changes of the Ach fractions, and seem to be of great significance at the initial stages of memory formation. PMID- 6273469 TI - Parainfluenza virus infections in the Cirencester Survey: seasonal and other characteristics. AB - Parainfluenza viruses were isolated 165 times during 14 years surveillance of the illnesses of a general practice population of around 3700. Type 1 isolations numbered 57, type 2 isolations 22 and type 3 isolations 86, representing annual rates of 33, 13 and 50 infections respectively per 10000 of population. Type 4 parainfluenza virus was not isolated. Three major classes of illness gave the following rates: sore throats (Throats) nine, acute febrile respiratory diseases (FRD) 23, acute non-febrile respiratory diseases (non-FRD) 71. The illnesses caused by the three types isolated were similar. Type 1 infections were most abundant in November and type 2 infections in December, and only 11.4% of these types were isolated in the warm semester April through September. Type 3 infections were seasonally bi-modal, with a winter peak in January and an even greater prevalence (66% of the total) in the warm semester. Type 3 infections in the warmer months and in the later years of the Survey were usually more severe. Type 3 virus may therefore be heterogeneous, one subtype possessing and the other lacking the genetic mechanism of "cold-season' prevalence. Geographical discontinuity between summer and winter isolation strengthens the case for the existence of the two subtypes of type 3 parainfluenza virus. Type 3 infections caused the majority of the infections in very young infants. Type 2 infections were widely distributed at all ages. Females were attacked more often than males: type 1, 68.4%; type 2, 63.6%; type 3, 53.5%. Type 3 infections in males outnumbered those in females up to 60 years of age, whereas female predominance became apparent in types 1 and 2 before 10 years of age. All types were widely and sparsely distributed, areas of prevalence changing from year to year. Recurrences occurred only twice, both with type 3 infections. Six persons suffered both a type 1 and a type 3 infection, and one person suffered both a type 2 and a type 3 infection. PMID- 6273470 TI - Use of cell differentiation effectors to select a human B lymphoblastoid cell line enriched in C3b receptors. AB - A study was undertaken to establish conditions of growth to increase C3b receptor synthesis on a human B lymphoblastoid cell line (Raji) by use of cell differentiation effectors. It appears that whereas two polar compounds HMBA (2mM) and Me2SO(2%) have no or little effect, 5 BrdU (30microM) and db cAMP (5 x10(-4) M) are able to increase in 48 h and 36 h respectively the synthesis of C3b receptor on Raji cell surface. These two compounds help to select a variant in which 100% of cells have C3b receptors with a high density of receptors per cell. The mechanism of BrdU action on the regulation of C3b receptor synthesis is discussed. PMID- 6273471 TI - Rapid microassays for the measurement of superoxide and hydrogen peroxide production by macrophages in culture using an automatic enzyme immunoassay reader. AB - Two simple semiautomated microassays for the measurement of superoxide (O-2) and hydrogen peroxide (H2O2) production by cultured macrophages (MPs) are described. The measurement of O-2 is based on the reduction of ferricytochrome c as assayed by the increase in its absorbance at 550 nm. Quantitation of H2O2 is based on the horseradish peroxidase (HRPO)-dependent oxidation of phenol red which is assayed by its increased absorbance at 600 nm. MPs are cultured in monolayers in 96-well flat-bottom tissue culture plates and covered with 100 mul amounts per well of either a ferricytochrome c solution containing phenol red and HRPO. Following the addition of an agent eliciting an oxidative burst (OB) and incubation of the plates at 37 degrees C for various time intervals, the changes in the absorbance of ferricytochrome c and phenol red, respectively, are measured directly in the wells of the tissue culture plates with the cells in situ, by using an automatic 8-channel photometer which reads absorbances vertically through individual wells. This instrument, which was originally designed for reading enzyme immunoassays in microtitration plates, can be easily adapted for use in the above test, when fitted with interference filters with wave lengths of 550 nm (for the assay of O-2) and 600 nm (for the assay of H2O2). The principal advantages of this techniques are: the ability to perform the assays directly in the culture plates with cells in situ; the small amounts of cells and reagents needed; its sensitivity and reproducibility; the ease with which kinetic experiments can be done; the large number of samples which can be tested in parallel, and especially the speed and convenience offered by the automated reading and printout of absorbance values. PMID- 6273472 TI - Use of cryopreserved lymphocytes for the indirect leukocyte migration inhibition assay. AB - Production of leukocyte migration inhibitory factor (LIF) by fresh and cryopreserved lymphocytes from the same donors was detected by indirect leukocyte migration inhibition (LMI) assay. The same results were obtained when fresh and frozen lymphocytes were tested in parallel. This indicates that cryopreservation does not impair the ability of lymphocytes to produce LIF. PMID- 6273473 TI - Patterns of shedding of myxoviruses and paramyxoviruses in children. AB - In the Houston Family Study, young children were cultured for virus weekly or biweekly and during acute respiratory illnesses. The interval between the onset of illness and positive culture was examined for 179 infections during 1975-1979. In week 1 after onset, 73%, 73%, and 66% of cultures were positive for influenza A virus, respiratory syncytial virus (RSV), and parainfluenza virus type 3, respectively. Pooled data from influenza B virus infections in 1977 and 1980 showed that 73% of cultures were positive in week 1. Influenza A virus in week 2 or RSV in weeks 2 and 3 was isolated from very few children. However, 37% of cultures were positive for influenza B virus during week 2, and 17% of cultures were still positive for parainfluenza virus type 3 during week 3. Shedding of parainfluenza virus type 3 on days 29-38 was also observed. Parainfluenza virus type 3, RSV, and influenza A virus were isolated up to six days before the onset of illness. PMID- 6273474 TI - Travelers' diarrhea in panamanian tourists in Mexico. AB - To determine whether residents of developing countries are unlikely to acquire travelers' diarrhea, 64 Panamanians of widely divergent socioeconomic strata were studied during a 15-day tour through Mexico. Twenty-three (36%) tourists experienced 27 episodes of travelers' diarrhea that were caused by seven different pathogens. The most commonly identified etiologic agents were rotavirus (26%), Norwalk virus (15%), and Campylobacter fetus (11%), whereas enterotoxigenic Escherichia coli was not frequently associated with travelers' diarrhea. Acquisition of travelers' diarrhea was correlated directly with high socioeconomic status. Varying levels of immunity to enteropathogens that are endemic in Panama may explain the different isolation rates of pathogens. PMID- 6273476 TI - Cytomegalovirus vaccines. PMID- 6273475 TI - Differences in neurovirulence among isolates of Herpes simplex virus types 1 and 2 in mice using four routes of infection. AB - Differences in neurovirulence between herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2) were investigated using recent clinical isolates and laboratory passaged strains in intravaginal, intranasal, intraperitoneal, and intracerebral infections of mice. The HSV-2 isolates caused higher death rates in all four infections. No differences in death rate were observed between recent and passaged isolates of either HSV-1 or HSV-2. After intravaginal inoculation, HSV-1 isolates replicated to higher titers in the vaginal mucosa, but HSV-2 isolates produced a higher death rate and a greater frequency of latent infection in lumbosacral ganglia of surviving animals. After intranasal inoculation, HSV-2 isolates again produced a higher death rate, but the frequency of latent infection in trigeminal ganglia was higher with HSV-1 isolates. The data suggest that the HSV-2 isolates have an enhanced capacity to enter and replicate in the central nervous system of mice but that latency is influenced by both virus type and route of inoculation. PMID- 6273477 TI - Nonenteric sources of rotavirus in acute diarrhea. PMID- 6273478 TI - [Cytochrome P-450-linked mixed function oxidase systems (monooxygenases) of microsomal and mitochondrial types and their substrate specificities (author's transl)]. PMID- 6273479 TI - [Vanadium and Na+, K+-ATPase (author's transl)]. PMID- 6273480 TI - [Erythrocyte membrane abnormalities in generalized membrane disorder (author's transl)]. PMID- 6273481 TI - [Beta 1-, beta 2-effects of ritrodine and terbutaline in the treatment of preterm labor (author's transl)]. AB - beta 2-Stimulants are at present the most effective tocolytic agents. The aim of the study was to evaluate the beta 1- . beta 2-effects of terbutaline and ritodrine in the treatment of preterm labor. 1) Terbutaline, administered intravenously at a rate of 0.4-1.2 microgram/min, and ritodrine also at a rate of 50-300 microgram/min, effectively inhibited uterine activity during 180 minutes. Among the patients there was no difference in responses between those who received terbutaline and ritodrine. 2) The levels of c-AMP and c-GMP in the blood after administration of terbutaline or ritodrine increased and showed dose response. The levels of c-AMP during a 180-minute infusion increased from 15 to 27 pmol/ml. 3) The action of c-AMP related substances on the circulatory systems was manifested as the dose-response of beta 1 action to the maternal blood pressure, maternal and fetal heart rate. Both beta 2-stimulants produced a tolerable changes in maternal heart rate and blood pressure, no significant changes during the infusion. PMID- 6273482 TI - [Effects of gonadotropin and LHRH on the cAMP levels, adenylate cyclase and cyclic nucleotide phosphodiesterase activities in the endocrine organs of the female pubertal rabbits (author's transl)]. AB - Effects of gonadotropin and LHRH on the cAMP levels, adenylate cyclase and cyclic nucleotide phosphodiesterase activities of the pineal, hypothalamus, pituitary, adrenal and ovary of the female pubertal rabbits were examined, and serum FSH, LH, estradiol and progesterone were measured. Gonadotropin and LHRH were injected intravenously five days, and 16 hours after the last administration, experiments were performed. The cAMP levels in the endocrine organs of the female pubertal rabbits were higher than adults. Gonadotropin and LHRH administration decreased the cAMP levels, i.e. draw to the levels of adults, in almost all endocrine organs. In pre-pubertal period (5-7 weeks of age), cAMP responses of hypothalamus and pituitary to gonadotropin or LHRH were significant, whereas responses of adrenal and ovary were slight and not significant. In early and mid-pubertal period (9-14 weeks of age), changes of the cAMP levels in the hypothalamus and pituitary were slight than pre-pubertal period, but in ovary, remarkable change of the cAMP levels were observed. The adenylate cyclase activities of all endocrine organs were not changed and the phosphodiesterase activities were increased by gonadotropin or LHRH administration. Serum FSH, LH estradiol and progesterone were increased with age. The most remarkable increases were occurred to the serum FSH level of pre-pubertal rabbits (7 weeks of age) after gonadotropin or LHRH administration, and serum progesterone of mid-pubertal rabbits (14 weeks of age) after gonadotropin administration. PMID- 6273483 TI - [DNP-peptidase activity in the rabbit ovary (author's transl)]. AB - To examine the possible roles of collagenolytic enzymes in the follicular wall at ovulation, we measured in rabbit ovaries collagenolytic enzymes by using synthetic substrates, DNP-Pro-Gln-Gly-Ile-Ala-Gly-Gln-D-Arg OH supposed to be specific for vertebrate collagenase. Our results clearly demonstrated that DNP peptidase activity existed, with its optimal pH7.6-pH7.8 in rabbit ovarian follicles. DNP-peptidase activity steadily increased for nine hours following the hCG injection. After nine hours of enzyme activity decreased sharply until about the tenth hour, at which time ovulation had occurred. This pattern of enzyme activities indicates that the DNP-peptidase was consumed for degradation of collagen fibers. After collagen fibers have been completely degraded, this enzyme was increased again. These changes of DNP-peptidase activities at ovulation were similar to cathepsin B1 activities with its optimal pH6.0. It is concluded that DNP-peptidase could be one of ovulatory enzymes in the rabbit. PMID- 6273484 TI - [Studies on prolyl hydroxylase activity, hydroxyproline content and solubility of collagen in the human uterus during pregnancy, delivery and postpartum involution (author's transl)]. AB - Prolyl hydroxylase activity, hydroxyproline content and solubility of collagen in the human pregnant and post-partum uterus were studied. The results were as follows: 1. Prolyl hydroxylase activity in the uterine cervix was slightly elevated during pregnancy, and the highest level was observed on the 4th day post partum. On the other hand, in the uterine body the highest activity was observed immediately after delivery. 2. Hydroxyproline content in the pregnant uterine cervix was slightly less than that of nonpregnant control and the lowest level was observed immediately after delivery, but on the 4th day post-partum it increased slightly. In the uterine body, there were no remarkable changes in content during pregnancy, but immediately after delivery it decreased remarkably. 3. Pepsin-soluble collagen in the uterine cervix increased significantly from an early pregnancy to immediately after delivery compared with nonpregnant control, but on the 4th day post-partum it decreased significantly compared with that immediately after delivery. In the uterine body, no significant changes in solubility were observed throughout pregnancy. 4. Comparing these data obtained on the cervix and the body at different stages of nonpregnancy, pregnancy and post-partum, enzyme activities in the body were always higher. Hydroxyproline content in the body of nonpregnant uterus showed a lower value than that in the cervix, but no remarkable differences were found between the cervix and the body throughout pregnancy and immediately after delivery. The significant decrease in pepsin-soluble collagen in the body was observed immediately after delivery. PMID- 6273486 TI - Cyclic nucleotide phosphodiesterases in normal and malignant human tissues. AB - Biopsy samples were obtained from tumor tissue and surrounding normal tissue from the same organs of the same patients in 25 patients. Adequate studies could be performed on all parameters in 18 tissue pairs: 12 malignant melanomas, 4 sarcomas, and 2 others. Significantly higher levels of cAMP low affinity phosphodiesterases were found in the tumors as compared to the controls. Similar differences were seen in the high affinity cAMP phosphodiesterases, but these were of borderline significance. High affinity cGMP phosphodiesterase levels did not differ significantly between normal and neoplastic tissue samples. Low affinity cGMP phosphodiesterases were not detectable in either tissues. These studies together with earlier experimental findings suggest possible auxiliary therapeutic trials with phosphodiesterase inhibitors. PMID- 6273485 TI - [Hormonal pituitary functions of women with increasing age (author's transl)]. AB - Pituitary functioning was observed endocrinologically in women in mature, climacteric and senile periods, and also patients after ovariectomy of both sides. Serum FSH and LH levels, especially in the former, showed a tendency towards increasing from the climacterium (45-49 years) up to the age of 60-64, and thereafter remained constant in a defined level. Serum PRL, ACTH and GH levels had a tendency to decrease after menopause. Serum TSH levels decreased from the age of 60-64 years. The change of responses for LH-RH tests with advancing age was found to be similar to that of serum gonadotropin levels. On the other hand, in the changes of response of PRL and TSH to TRH test with increasing age, there was a reverse tendency to that of gonadotropin to LH-RH. In ovariectomized women who still had a menstrual cycle, there was a great increase in serum FSH and LH after the operation, as against those who had spent 3 years and 10 months, or 4 years and 4 months after menopause, a slight or little increase respectively. Meanwhile, the LH response to LH-RH was higher in the premenopausal ovariectomized women than that of the post-menopausal unovariectomized women. PMID- 6273487 TI - Epidemiologic, etiologic and immunologic aspects of nasopharyngeal carcinoma [NPC]. PMID- 6273488 TI - Diagnosis of lung cancer by transbronchial lung biopsy and a comparison between this method with the sputum and bronchial washing cytology in Ramathibodi hospital. PMID- 6273489 TI - Immunohistochemical localization of cyclic AMP in the developing rodent secondary palate. AB - During development of the mammalian secondary palate, medial-edge epithelia (MEE) from apposing palatal shelves adhere and undergo autolysis allowing palatal mesenchymal regions to unite. In a prior study (Greene & Pratt, 1979), we reported a transient increase in levels of cyclic AMP (cAMP) in the mouse and rat palate during epithelial adhesion and cell death. The objective of this study was to examine the distribution of cyclic AMP in the developing rodent secondary palate using immunohistochemistry to localize cyclic AMP. Staining for cAMP was observed in the epithelium just prior to and during epithelial fusion (day 16 in the rat; day 14 in the mouse). Cyclic AMP was distributed throughout the epithelial cytoplasm whereas no staining was seen in nuclei. Epithelial staining for cAMP was faint or absent 24 and 48 h prior to epithelial contact. Mesenchymal staining for cAMP was minimal and associated with the plasma membrane at all stages studied. These results demonstrate that elevated levels of cAMP in the rat and mouse palate during epithelial adhesion and cell death are mainly due to increases of the nucleotide in palatal epithelium. This observation suggests that a transient increase in epithelial cAMP may play a role in palatal epithelial differentiation. PMID- 6273490 TI - Application of a live varicella vaccine in children with acute leukemia or nephrotic syndrome. PMID- 6273491 TI - Plasma renin activity and cyclic adenosine monophosphate in orthostatic hypotension of spinal transections. PMID- 6273492 TI - Peritoneoscopic guided aspiration cytology in the diagnosis of liver malignancy. PMID- 6273493 TI - Depressed cellular immunity in hepatocellular carcinoma patients. PMID- 6273494 TI - Cyclic AMP-stimulated chloride fluxes in dialyzed barnacle muscle fibers. AB - Unidirectional chloride efflux and influx were studied in giant barnacle muscle fibers that were internally dialyzed. When cyclic 3'5'-adenosine monophosphate (cAMP) was included in the dialysis fluid, both unidirectional fluxes were stimulated by about the same amount. This stimulation was not associated with measurable changes either in membrane electrical conductance or with net movements of chloride. The stimulation required the trans-side presence of chloride. The stimulated flux was inhibited by the sulfonic acid stilbene derivatives 4-acetamido-4'-isothiocyanostilbene-2',2'-disulfonate (SITS) and 4,4' diisothiocyanostilbene-2,2'-disulfonate (DIDS) or by furosemide. When cAMP was presented in high concentrations (10-5 M), the effect on chloride fluxes was characterized by a desensitization phenomenon. This desensitization was not the result of an increased amount of phosphodiesterase activity, but may be related to ATP and/or intracellular calcium levels. These results further support the hypothesis that the barnacle sarcolemma possesses a specialized chloride transport mechanism that largely engages in Cl-Cl exchange under conditions of normal intracellular pH. PMID- 6273495 TI - Membrane-bound ATP fuels the Na/K pump. Studies on membrane-bound glycolytic enzymes on inside-out vesicles from human red cell membranes. AB - ATP stimulates Na transport into inside-out vesicles (IOVs) made from human red cell membranes; strophanthidin inhibits the ATP-stimulated transport. The substrates for glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and phosphoglycerate kinase (PGK) (glycolytic enzymes bound to the cytoplasmic surface of the red cell membrane) also stimulate Na transport into IOVs without added ATP. The elution of GAPDH from the membranes prevents the stimulation by the substrates, but not by exogenous ATP. Hexokinase plus glucose (agents that promote breakdown of ATP) prevent stimulation of Na transport by exogenous ATP but not by the substrates for GAPDH and PGK. [32P]orthophosphate is incorporated into a membrane-bound organic phosphate compound shown chromatographically to be ATP. The level of membrane-bound ATP is decreased when Na is added, and this decrease is inhibited by strophanthidin. When further synthesis of [32P]ATP is blocked by the addition of unlabeled orthophosphate, all of the membrane-bound [32P]ATP is dissipated by the addition of Na. From these observations it was concluded that membrane-bound glycolytic enzymes synthesize ATP and deposit it in a membrane-associated compartment from which it is used by the Na/K pump. PMID- 6273496 TI - Virus detection by nucleic acid hybridization: examination of normal and ALS tissues for the presence of poliovirus. AB - A nucleic acid hybridization assay was developed as a sensitive assay for the presence of poliovirus RNA in human tissue. The assay could detect the presence of an average of one poliovirus per 200 cells. A method for determining the extent of degradation of the tissue RNA was developed and used to show that a significant fraction of human central nervous system (CNS) autopsy material contains highly degraded RNA which is unsuitable for hybridization studies. A total of 15 different control and amyotrophic lateral sclerosis tissues were assayed for the presence of poliovirus-like RNA. Virus RNA was detected in one of the control tissues and in none of the ALS tissues. PMID- 6273497 TI - Thermal inactivation studies of a coronavirus, transmissible gastroenteritis virus. AB - The thermolability of transmissible gastroenteritis virus (TGEV) was studied between 31 and 55 degrees C using two different strains. The loss of infectivity followed first order kinetics except at the highest temperature. The values of the thermodynamic parameters indicated that the mechanisms involved above and below 45 degrees C are clearly distinct. The rates of inactivation were greater at alkaline than at neutral pH, yet the name of the reaction appeared unchanged. Using four independent stocks of mutagenized virus, we failed to select thermal resistant mutants by survivor selection at 38 degrees C. In contrast, thermal resistant mutants were consistently obtained at 54 degrees C. However, the latter did not show any increased stability at 38 degrees C, confirming the fact that a different inactivation process takes place at high and at physiological temperatures. PMID- 6273498 TI - Excretion of polyamines from baby hamster kidney cells (BHK-21/C13: effect of infection with Herpes Simplex Virus Type 1. AB - Confluent monolayers of BHK-21/C13 cells excreted increasing amounts of polyamines into the extracellular medium with time. Excretion was specificity of spermidine and was not the result of cell death or lysis. Herpes simplex virus type 1 (HSV-1) completely prevented excretion of polyamines from 2 h after infection of the cells. The virus specificity inhibited the release of free spermidine but not of conjugated polyamines. PMID- 6273499 TI - Effects of oral treatment with acyclovir and bromovinyldeoxyuridine on the establishment of maintenance of latent herpes simplex virus infection in mice. AB - Mice infected with herpes simplex virus (HSV) were treated (separately) with the nucleoside analogues acyclovir or bromovinyldeoxyuridine by incorporating the drugs in the drinking water. This method of treatment was found to be effective for both drugs and compared favourably with intraperitoneal injection. Prompt treatment with either compound could prevent the establishment of latent infections but latent infections once established were intractable using prolonged courses of oral administration. PMID- 6273500 TI - Partial exclusion of bacteriophage T2 by bacteriophage T4: induction of early enzymes by excluded T2. AB - In crosses between bacteriophages T2 and T4 most early genes of T2 are partially excluded from the progeny. Six genes of T4 affect the exclusion of six corresponding exclusion-sensitive sites in T2, each gene being specific for the exclusion of one site. Mutants of T4 in these genes have been isolated (ex mutants). The induction of the gene product (deoxycytidine triphosphate nucleotidohydrolase, dCTPase) of the strongly excluded T2 gene 56 was determined. The dCTPase was induced in the presence of the replication inhibitor oxolinic acid to prevent possible artefacts from unequal replicaton rates of T2 and T4. The rate of T2 dCTPase induction was 30% of the control in T2 X T4 infections in which all six exclusion-sensitive sites were excluded and was 57% in infections in which the sites except 56 were excluded. The dCTPase induction was 67% of the control with exclusion of the 56 site only and equalled the control when no T2 site was excluded. Inhibition of dCTPase induction under conditions of exclusion is partly due to exclusion of neighboring sites and partly due to exclusion of the 56 site. PMID- 6273501 TI - Composition and topography of structural polypeptides of bovine rotavirus. AB - The composition and topography of the structural polypeptides of bovine rotavirus was studied by polyacrylamide gel electrophoresis of radioactively labelled virus grown in LLC-MK2 cells and by lactoperoxidase-catalysed iodination of single- and double- capsid particles. Bovine rotavirus was found to possess at least six structural polypeptides, three of them associated with the inner capsid (p102K, p91K and p45K) and the others with the outer capsid (p84K, p37K and p34K). The most abundant polypeptide of the inner capsid was p45K, which accounted for approx, 80% of the protein mass, followed by p91K (approx, 20% of the protein mass) and p102K (approx. 1% of the protein mass). Polypeptide 45K is not readily available for iodination, indicating that it is partially covered by p91K, which is the most exposed polypeptide of the inner capsid. The number of polypeptide molecules per single capsid particle (calculated on the basis of an RNA content of 16%) was estimated to be approx. 6 molecules of p102K, 140 of p91K and 989 of p45K. The stoichiometry and degree of exposure of outer capsid polypeptides was more difficult to establish, even when it appears that p84K and p34K are the most exposed components. PMID- 6273502 TI - Multiple genetic changes can occur in the oral poliovaccines upon replication in humans. AB - Poliovirus isolates of serotypes 2 and 3 from patients whose paralytic poliomyelitis cases were classified as oral vaccine-associated were analysed by oligonucleotide mapping of the virus genomes and by polyacrylamide gel electrophoresis of the virus proteins. Oligonucleotide maps of all isolates were similar to the maps of the corresponding oral vaccine strain. No two isolates gave identical maps. Most maps differed from that of the vaccine strain by at least one oligonucleotide spot. Maps of some isolates revealed numerous differences, indicating that multiple (greater than 100) genetic changes had occurred in the vaccine virus genomes during replication in one or two individuals. In contrast, maps of some neural tissue isolates showed minimal differences from the reference vaccine maps, raising the possibility that neurovirulence may be restored by a small number of genetic changes. For many isolates, changes were also detected in the mobilities of processing rates of the virus proteins. PMID- 6273503 TI - Biochemical transformation of mouse cells by varicella-zoster virus. AB - Mouse L cells lacking the enzyme thymidine kinase (Ltk-) were infected with varicella-zoster virus (VZV). Even though virus did not replicate in Ltk- cells, the presence of virus antigen could be observed by use of an anti-complement immunofluorescent technique at 4 h post-infection and the VZV-specific thymidine kinase could be detected in VZV-infected Ltk- cells. Ltk-cells were converted to a tk+ phenotype (Ltk+) by infection with cell-associated VZV. Clones possessing the ability to grow in selective medium were isolated and cultured successfully for more than 20 passages. One of the clones grew very slowly, but other clones showed almost the same growth rate as that of the parental Ltk- cells. The chromosome analyses of Ltk- cells and transformed cells revealed that the isolated clones were of mouse origin. VZV-specific antigen could be detected in the nuclei of Ltk+ cell clones by an immunofluorescent test, while tk activity was greatly enhanced in extracts prepared from transformed cells and its activity was neutralized by hyperimmune serum against VZV. PMID- 6273504 TI - Electron spin resonance study of the copper(II) complexes of human and dog serum albumins abd some peptide analogs. AB - Electron spin resonance spectra of the first Cu(II) complexes of human serum albumin, dog serum albumin, L-aspartyl-L-histidine N-methylamide and glycyl-gly cyl-L-histidine N-methylamide have been studied using isotopically pure 65Cu in its chloride form. At 77 degrees K, the esr spectra of Cu(II) complex of human serum albumin exhibited only one form of esr signal between pH 6.5 and 11. No intermediate forms were detected. The presence of an equally spaced nine-line superhyperfine structure with spacing approximately 15 G indicated considerable covalent bonding between Cu(II) and four nitrogen atoms derived from the protein. The esr spectrum form of Cu(II) bound to human serum albumin detected at neutral pH would be consistent with the participation of four nitrogens from the alpha NH2 group, two peptide groups, and the imidazole group of a histidine residue. In contrast, the esr spectra of Vu(II)-dog serum albumin complex showed a transition from a low pH form to a high pH form as the pH was increased to 9.5. These spectral changes were found to be reversible upon lowering the pH. Ligand superhyperfine splitting in the low pH form of the esr signal of Cu(II)-dog albumin were not resolved. The distinct pH dependence of the esr signals observed in human and dog serum albumin complexes could be correlated to their respective optical spectra changes as a function of pH. At room temperature and in the pH range between 6 and 11, the esr spectra of Cu(II) complexes of L-aspartyl-L alanyl-L-histidine N-methylamide and glycyl-glycyl-L-histidine N-methylamide exhibited a well-resolved nine-line superhyperfine structure indicating metal coordination with four equivalent nitrogen atoms of peptide. PMID- 6273505 TI - Catalysis of the disproportionation of superoxide by metalloporphyrins. III. AB - An assay based on the use of an oxygen electrode has been used to determine the superoxide dismutase-type activity of several metalloporphyrins. Both the iron(III) and manganese(III) derivatives of tetrakis-(4-N-methylpyridyl)porphine were shown to have comparable activity, about 2-3% of that of the enzyme. Comparisons are made between this assay method and the more standard one which depends on the conversion of nitroblue tetrazolium to formazan. PMID- 6273506 TI - Subacute arsenic neuropathy: clinical and electrophysiological observations. AB - Two patients with subacute arsenic neuropathy are described and the results of serial conduction velocity determinations from the very early stages of the illness to partial recovery are reported. Sensory and motor deficits were maximal within four weeks of the estimated time of exposure. Recovery was slow, with only partial improvement of the neurological deficits two years after onset of the illness. Progressive slowing of motor conduction velocity was observed in the first three months followed by a gradual increase in velocity thereafter. The possible mechanisms underlying the temporal profile of the electrophysiological changes are considered. The need for initiating chelation therapy before the development of the neuropathy is emphasised. PMID- 6273508 TI - Relative preservation of lumbrical versus thenar motor fibres in neurogenic disorders. AB - At late stages of median nerve entrapment in the carpal tunnel there may be total denervation of the thenar muscles. Surface electrodes over the thenar endplate zone may record an initial and predominantly positive "M" response to supramaximal median nerve stimulation. By a combination of techniques, it has been established that this surface positive response originates from surviving lumbrical muscles innervated by the median nerve. Their relative preservation may be related to their location in the median nerve. Intraoperative investigations have shown that the thenar motor fibres are primarily located in the ventral lateral part of the median nerve whereas lumbrical motor fibres are in a more dorsal location and hence are probably better protected against the flexor retinaculum. This relative preservation of lumbrical motor fibres has been observed in other peripheral neuropathies and motor neuron disease, but not in median nerve regeneration of the following nerve transection. The importance of the observation lies in the more precise localisation voltage source for "M" response in terminal or near-terminal carpal tunnel median nerve entrapments and avoidance thereby of possible misinterpretation of electrophysiological observations in this most common nerve entrapment syndrome. PMID- 6273507 TI - Megaoesophagus due to acrylamide neuropathy. AB - Greyhound dogs exposed to oral acrylamide for a period of eight weeks developed a sensorimotor peripheral neuropathy that had many features in common with acrylamide neuropathy seen in other species. Most of the animals also developed the clinical and radiological features of megaoesophagus. The association of neuropathy and megaoesophagus suggests that an axonopathy of the vagus may be an aetiological factor in this disorder. PMID- 6273509 TI - Characteristics of the cyclic AMP-phosphodiesterase activator in human brain tumours. AB - The levels of the cyclic adenosine 3',5'-monophosphate (cyclic AMP) phosphodiesterase (PDE) and the biochemical properties of its endogenous protein activator (PDEA) obtained from the human brain cortex and from different types of human cerebral tumours have been evaluated. The effects of the various PDEAs were studied measuring the activation of an activator-depleted cyclic AMP-PDE prepared from a normal brain cortex. The PDEA, obtained from normal and pathological tissues, did not change the affinity of the purified PDE for cyclic AMP, while it increased the Vmax of the enzyme. On the other hand, a cross-activation study showed that the PDEA lacked tissue specificity and was present in the tissue in excess over the enzyme. The levels of cyclic AMP-PDE and PDEA were much higher in normal than in tumoural tissues. The enzyme activity decreased in cerebral tumours more markedly than the protein activator. This biochemical pattern was more evident in the tumours of glial origin which are the most malignant. PMID- 6273510 TI - Regularity and differentiation within the structure of brain postsynaptic densities. AB - Postsynaptic densities (PSDs) isolated from rat forebrain were examined in samples prepared for electron microscopy by negative staining. Two structurally distinct areas could be distinguished within each individual PSD, a peripheral zone composed of a planar array of spherical subunits with a mean diameter of 18 nm and one or more islands of fine granular material enclosed by the subunits. These enclosures correspond in distribution and size to 'holes' which are known to occur in PSDs in intact brain tissue. The spherical subunits can occur singly in the isolated PSD preparations showing that the structure of each individual subunit does not depend upon its interaction with others. These observations indicate that the PSD structure has both regular and differentiated features. These are considered in relation to the possible roles of the PSD in defining the postsynaptic locus and mediating the postsynaptic events of synaptic transmission. PMID- 6273511 TI - Tc-99m DMSA renal uptake: influence of biochemical and physiologic factors. AB - Thirty-eight female Sprague-Dawley rats were studied to determine the effects of (a) tubular blockade and (b) commonly encountered changes in hydration and acid base balance, on the urinary excretion and renal localization of Tc-99m dimercaptosuccinic acid (DMSA). Ten additional rats were studied to quantitate the in vivo protein binding of Tc-99m DMSA, and a final group of 12 animals was used to quantitate DMSA distribution in animals with diminished functional renal mass. Both osmotic diuresis and dehydration by water deprivation for 24 hr resulted in a plasma clearance of DMSA slower than in control animals. Acid-base imbalances significantly affected the renal accumulation of DMSA, and acidosis was associated with markedly increased background due to increased liver accumulation. The protein-bound portion of Tc-99m DMSA in the plasma was high, reaching 89% within the first 5 min, and rising very slightly (n.s.) ith time. The unbound portion of DMSA had a plasma clearance slightly higher than the GFR. Ablation of large amounts of renal tissue, resulting in significant decreases in GFR. Ablation of large amounts of renal tissue, resulting in significant decreases in GFR, did not significantly affect the renal localization of DMSA in the intact portions of the kidneys. These data demonstrate that commonly encountered changes in acid-base balance and hydration will significantly alter the biologic distribution of Tc-99m DMSA. These factors should be controlled when carrying out clinical studies. PMID- 6273512 TI - Does the corticoadrenal adenoma with "pre-Cushing's syndrome" exist? AB - An adrenal tumor was discovered fortuitously in a patient with no clinical features of Cushing's syndrome. On adrenal imaging, there was good uptake in the nodule but no visualization of the contralateral adrenal. The latter was seen, however, in a second scan performed under ACTH treatment. In the hormone assessment, basal cortisol and 17-hydroxycorticoids were normal and cortisol diurnal variation was near normal, but a dexamethasone suppression test and ACTH responses to metyrapone and insulin hypoglycemia were abnormal. Eight months after excision of a spongiocytic-type adenoma, the remaining adrenal was visible on scintigram and the hormonal tests were normal. This pattern suggests that the clinical Cushing's syndrome but enough to suppress partially ACTH and, consequently, visualization of the contralateral gland. PMID- 6273513 TI - Decreased accumulation of isopropyl-lodoamphetamine (I-123) in brain tumors. PMID- 6273514 TI - Renal acid, urinary cyclic AMP, and hydroxyproline excretion as affected by level of protein, sulfur amino acid, and phosphorus intake. AB - Two 51-day human studies were conducted to investigate the effects of level of protein and phosphorus intake on the various components of renal acid excretion and on urinary sulfate, cyclic AMP and hydroxyproline; the role of the sulfur amino acids (Saa) of the protein was also evaluated. Dietary treatments included: 1) a 50 g protein diet; 2) a 150 g protein diet; and 3) a 50 g protein diet plus Saa to equal that of the 150 g protein diet, each given at 2 levels of phosphorus (1010 and 2525 mg). Calcium intake was 500 mg. Subjects were 16 young adult males. The results are discussed in relationship to calcium data previously reported (1, 2). Changes in renal acid and calcium excretion are not directly related for these reasons: a) the Saa accounted for all of the protein-induced increase in urinary sulfate and acid but for only 43% of the increase in urinary calcium and b) the acid phosphate supplement decreased urinary calcium but increased total acid excretion. The phosphorus supplement increased cyclic AMP but not hydroxyproline excretion. In fact, protein and Saa caused increases in hydroxyproline that were greatly reduced by the phosphorus supplement. Increases in urinary hydroxyproline and calcium were well correlated indicating that, at low calcium intakes, protein or Saa-induced increases in urinary calcium result in increased bone resorption which is reduced by the administration of phosphorus. PMID- 6273515 TI - Effects of tomato pomace and mixed-vegetable pomace on serum and liver cholesterol in rats. AB - The effects of tomato pomace and mixed-vegetable pomace, vegetable processing by products, on serum and liver cholesterol were compared against pectin, wheat bran, cellulose, and lignin. Male Sprague-Dawley rats (100 g) were fed high sucrose, semi-purified diets containing basal (0), 5% or 10% of the fiber source and either 0, 0.5% cholesterol or 0.5% cholesterol + 0.125% sodium cholate. After 28 days, there were highly significant differences (P less than 0.001) in serum and liver cholesterols due to fiber source, dietary cholesterol and the interaction. The effect of fiber source level was significant (P less than 0.05) for liver cholesterol only. Pectin groups had serum and liver cholesterol levels lower (P less than 0.05) than the others. At the 10% level, tomato pomace, wheat bran and lignin (Indulin AT) groups had serum cholesterols higher (P less than 0.05) than basal; the mixed-vegetable pomace and cellulose groups were not different from basal. Liver cholesterols of tomato pomace, mixed-vegetable pomace, wheat bran and lignin groups were not different at the 10% level, but were higher than the pectin, cellulose and basal groups (P less than 0.05). In a second experiment, serum cholesterols of rats fed pectin were lower (P less than 0.05) than for those fed cellulose or two types of lignin (Indulin AT, Reax 27). PMID- 6273516 TI - [A functional test of the salivary gland using 99mTc-pertechnetate--diagnostic values of the intraoral radioactivities (author's transl)]. PMID- 6273517 TI - Clinical, virologic, and serologic evidence of Epstein-Barr virus infection in association with childhood pneumonia. AB - To explore the association of Epstein-Barr virus infection with childhood pneumonia we studied two patients whose mononucleosis-like illnesses were accompanied by pneumonia; both had virologic and serologic evidence of current or recent EBV infection. We then analyzed the sera of 71 children (age range, 14 months to 9 years) with pulmonary infiltrates for the presence of four classes of antibody to EBV. Antibody responses consistent with current or recent EB virus infection were found in 15. Two children had IgM antibodies to the EBV viral antigen at titers greater than or equal to 1:160, indicating current infection, and all 15 patients had antibody to components of the early antigen complex, suggesting recent infection. A fourfold rise or drop in one or more EBV-specific antibody classes was noted in eight patients within 30 days following onset of clinical illness. Few patients had clinical features suggesting infectious mononucleosis. Eight of the 15 with serologic evidence of current or recent EBV infection also had clinical or serologic evidence of infection with another pathogen--bacterial, viral, or mycoplasmal. Thus, in childhood pneumonia, EBV may be a primary, co-primary, or secondary pathogen; it may be reactivated in the course of infection with another agent, or possibly, by suppressing immune function, it may precipitate infection with some other organism. PMID- 6273518 TI - The syndrome of hyperostosis and hyperphosphatemia. AB - Six children, five girls and one boy, presented with recurrent episodes of swelling, pain, and tenderness of the long bones. On roentgenographic examination all had cortical hyperostosis of the affected areas. Serum phosphate concentration was persistently elevated, and calcium values were normal. Bone biopsy and histologic examination in three patients revealed periosteal new bone formation. The Ellsworth-Howard test was performed on three patients; all had a normal phosphaturic response and an increase in urinary c'AMP to exogenous PTH. The EDTA test, performed on one patient, demonstrated significant phosphaturic response, but a minimal drop in serum phosphate concentration. These findings suggest that the association of cortical hyperostosis and hyperphosphatemia is a distinct clinical entity, and that hyperphosphatemia results from decreased renal excretion of phosphate. PMID- 6273519 TI - Intravenous lipid emulsions and human neutrophil function. PMID- 6273520 TI - Lactose malabsorption following rotavirus infection in young children. PMID- 6273521 TI - The coexistence of Wilms' tumor and imperforate anus with rectourethral fistula. AB - Wilms' tumor or nephroblastoma is one of the more common abdominal neoplasms in childhood. Its association with aniridia, hemihypertrophy, and genitourinary anomalies is well recognized. We believe this is the first report of Wilms' tumor coexistent with imperforate anus and rectourethral fistula. PMID- 6273522 TI - Congenital mesoblastic nephroma. AB - Neonatal renal neoplasms are rare: most are congenital mesoblastic nephroma. In the past this neoplasm has been confused with Wilms' tumor, which in part may account for some of the more favorable prognosis ascribed to Wilms' tumor in children less than 1 yr of age. Congenital mesoblastic nephroma is usually a benign tumor. The recommended treatment is surgical excision. This is a report of two cases of congenital mesoblastic nephroma in infants who were treated successfully by nephrectomy. PMID- 6273523 TI - Immunolocalization of collagenase in periodontal disease. PMID- 6273524 TI - Cyclic nucleotide content in gingival tissue of smokers and non-smokers. PMID- 6273525 TI - Conditions of production and properties of the collagenolytic enzymes by two Bacillus strains from dental plaque. PMID- 6273526 TI - Synthesis, structure, and features of anilinium anilinomethanesulfonates. AB - Two series of anilinomethanesulfonate derivatives, the anilinium and p toluidinium anilinomethanesulfonates, were synthesized. They present interesting spectroscopic features and can be used as characteristic derivatives of the parent compounds. These series allow the easy study of the N--H stretching frequencies of the secondary amine whereas difficulties are encountered with the parent compounds due to the occurrence of O--H stretching bands in the same frequency range. All three compounds present a common fragmentation pattern in mass spectral analysis. Through these analyses, the salt structure of the anilinium anilinomethanesulfonates is demonstrated in opposition to a sulfonamide structure, giving further support to the salt structure of other series previously reported. PMID- 6273527 TI - Effects of very low concentrations of ouabain on contractile force of isolated guinea-pig, rabbit and cat atria and right ventricular papillary muscles: an interinstitutional study. PMID- 6273528 TI - Interactions of d-tubocurarine with the nicotinic acetylcholine receptor/channel molecule. AB - The interactions of d-tubocurarine (d-TC) with the ionic channel of the nicotinic acetylcholine receptor were studied by biochemical methods in Torpedo electric organ membranes and by electrophysiological methods on frog sciatic nerve sartorius muscle preparation. Torpedo membranes were treated with alpha bungarotoxin to inhibit the acetylcholine receptor sites, then binding of [3H]perhydrohistrionicotoxin to the ionic channel sites was studied and found to be inhibited by d-TC. At 37 degrees C, the Ki of d-TC was 10 microM, and at 22 degrees C it was 100 microM. The affinity of d-TC for the ionic channel sites relative to that of perhydrohistrionicotoxin was constant at temperatures from 2 20 degrees C, but increased at higher temperatures up to 37 degrees C. The peak endplate current amplitude was depressed with 1 to 2 microM d-TC in a voltage dependent manner, with considerable departure from linearity at 10 and 30 degrees C. The effect of d-TC on spontaneous miniature endplate currents was similar and slightly more potent. The time constant of endplate current decay was decreased by d-TC (1 and 2 microM) at temperatures of 10, 15 and 30 degrees C. The channel lifetime was reduced by d-TC, but channel conductance was unaffected. It is suggested that d-TC interacts with both the acetylcholine receptor sites as well as its ionic channel sites in closed and open conformations. PMID- 6273529 TI - Angiotensin converting enzyme inhibition in tissues from spontaneously hypertensive rats after treatment with captopril or MK-421. PMID- 6273530 TI - Voltage noise measurements across the pancreatic beta-cell membrane: calcium channel characteristics. AB - 1. Membrane potential fluctuations were measured in cells from mouse Islets of Langerhans identified as beta-cells by the characteristic pattern of electrical activity induced by 11 mM-D-glucose. 2. The membrane potential was controlled by adjusting the external potassium concentration, [K+]o, keeping the sum [Na+]o plus [K+]o constant. In the absence of glucose, when [K+]o is raised, the resulting depolarization is accompanied by a significant increase in voltage noise. 3 The amplitude and time course of the voltage noise were measured under various experimental conditions. The variance of the fluctuating voltage decreased monotonically along the depolarization induced by sudden increase in [K+]o, suggesting a monotonic reduction in the number of elementary events. 4. The frequency characteristics of the excess noise could be analysed as the sum of 1/f and 1/f2 components. While the 1/f component remained unaffected by the external application of 20mM-tetraethylammonium (TEA) and either 2 mM-Mn2+ or 2 mM-Co2+, the 1/f2 component was suppressed by both Mn2+ and Co2+. 5. The corner frequency, fc, of the 1/f2 component depended on membrane potential, which was adjusted by adjusting the [K+]o jump. These results support the idea that fc in these experiments is a measure of the channel relaxation. 6. Measurements of the input resistance in the frequency range from 0 to 25 Hz were used to obtain a rough estimate of the size of the channel conductance as 5 x 10(-12) omega (-1). PMID- 6273531 TI - Intracellular magnesium does not antagonize calcium-dependent acetylcholine secretion. AB - 1. The effects of intracellular application of Ca and Mg ions on evoked acetylcholine secretion at frog motor nerve terminals were studied. Ca and Mg were applied to the nerve-ending cytoplasm using liposomes as a vehicle. 2. Under conditions in which intracellular application of Ca produced many-fold increased in evoked acetylcholine release. 3. When Mg was applied to the nerve-ending cytoplasm concurrently with Ca, acetylcholine release was further increased above the level produced by introducing Ca alone. 4. The results suggest that intracellular Mg does not antagonize depolarization-secretion coupling and that antagonism of transmitter release by extracellular Mg occurs only at the external surface of the nerve ending. PMID- 6273532 TI - Calcium-mediated inactivation of the calcium conductance in caesium-loaded giant neurones of Aplysia californica. AB - 1. The intracellular potassium in giant neurones of Aplysia californica was replaced with caesium by a method utilizing the ionophore nystatin. Because caesium ions have low permeability through potassium channels outward currents during voltage-clamp depolarization were strongly curtailed after the caesium loading procedure and the subsequent wash-out of the ionophore. 2. The calcium current elicited by a test voltage-clamp depolarization (pulse 2) was depressed following the entry of calcium elicited by a prior depolarization (pulse 1). 3. The percentage depression of the test current was a linear function of the pulse 1 current-time integral, and thus appears to be related linearly to the amount of calcium carried into the cell during pulse 1. This linear relation was maintained when calcium entry was varied by changes in external calcium concentration, by altered pulse 1 amplitude and altered pulse 1 duration. Depression was substantially reduced by injection of EGTA, and by substitution of barium for extracellular calcium. 4. The calcium current was unaffected by prior hyperpolarization of the membrane or by prior depolarizations to about ECa. Depression of the current was not altered by addition of extracellular 50 mM-TEA or by a strong hyperpolarization between the conditioning and test pulses. 5. The rate relaxation of the inward current during a given depolarization depended on the rate of entry and accumulation of free calcium. Relaxation under a given command potential became slower when calcium was partially replaced with magnesium so as to produce a smaller calcium current, or when accumulation of intracellular free calcium was retarded by injected EGTA or by barium substitution for extracellular calcium. 6 Evidence is considered that accumulation of calcium ions at the cytoplasmic surface of the membrane leads to inactivation through an action upon the calcium conductance. Reduced driving force and intracellular surface-charge neutralization do not adequately account for the observed depression of the calcium current resulting from intracellular accumulation of calcium ions. PMID- 6273533 TI - A new interpretation of the pace-maker current in calf Purkinje fibres. AB - 1. The properties of the 'pace-maker' current iK2 of Purkinje fibres are investigated to verify whether its behaviour during voltage-clamp pulses is consistent with the view that iK2 is a K current deactivating on hyperpolarization in the range -50 to -100 mV. 2. During voltage-clamp pulses in the range positive to the apparent reversal potential (Erev), low concentrations of Cs depress the time-dependent current change without altering the time independent current component. The total current becomes more outward in Cs, which on the assumption that Cs only affects the iK1 and iK2 channels implies that iK2 is inward in the voltage range analysed. 3. Ba strongly reduces the time independent component due to iK1 and therefore limits the contribution of K depletion to the total current time course during hyperpolarizations. In the presence of barium a current reversal is not obtained even with large hyperpolarizations in normal Tyrode solution. If in Ba-containing solutions the external K concentration is raised from 3 up to 48 mM, iK2 greatly increases in the inward direction during hyperpolarizations in the range -51 to -101 mV, implying that it cannot be carried by K only. 4. In the presence of Ba, measurements of the membrane conductance due to iK2 indicate a channel-opening process during hyperpolarizations and a channel-closing process during depolarizations, in the K-concentration range analysed. 5. It is concluded that iK2 is not, as had been previously thought, a pure K current, but is rather an inward current activated during hyperpolarizations negative to about -50 mV. This provides further evidence for a possible identity between iK2 in Purkinje fibres and the current if in the SA node. PMID- 6273534 TI - A study of the ionic nature of the pace-maker current in calf Purkinje fibres. AB - 1. Properties of the pace-maker current (if) in Purkinje fibres were studied in the presence of Ba, which by partially blocking the iK1 channel reduces K depletion during hyperpolarizing voltage-clamp pulses, and eliminates the main cause of distortion in the current time course. 2. On raising the external potassium concentration (Kb), the if fully activated current--voltage relation (if(E)) increases in the inward direction. In the range 3--36 mM--Kb and negative to -50 mV the current is inward, and no cross-over is observed. 3. In normal conditions, the reversal potential (Ef) for if lies in the voltage region positive to -50 mV, and can be observed on lowering the external sodium concentration (Nab). Ef shifts to the negative direction when Nab is decreased. Slopes ranging between 29 and 35 mV/decade are found for Nernst plots of Ef against Nab. Changing Nab in the range 140--4.4 mM causes the if(E) relation to undergo a simple shift along the voltage axis, without significant change in its slope. 4. Ef also depends on Kb, as can be observed in low Nab (35 mM), and shifts to the positive direction by about 26 mV for every 10-fold change in Kb. The fully activated slope conductance increases when Kb is increased. 5. It is concluded that Na and K both participate in carrying if. The slope of the fully activated if(E) relation increases with Kb, but is unchanged in different Nab, indicating that the channel conductance depends on Kb, but not appreciably on Nab. PMID- 6273535 TI - The effects of ATP on the interactions between monovalent cations and the sodium pump in dialysed squid axons. AB - 1. The efflux of Na in dialysed axons of the squid has been used to monitor the sidedness of the interactions of the Na pump with Na(+) ions, K(+) ions and ATP. The axons were under conditions such that most of the Na efflux went through the Na pump by means of a complete cycle of ATP hydrolysis.2. With 310 mm-K(i) (+), 70 mm-Na(i) (+) and 10 mm-K(+) artificial sea water (ASW) more than 97% of the Na efflux was abolished by removal of ATP. The efflux of Na was stimulated by ATP with a K((1/2)) of about 200 mum. This is similar to the K((1/2)) of 150 mum found for the ATP dependence of a ouabain-sensitive Na,K-ATPase activity in membrane fragments isolated from squid optical nerves.3. A 100-fold reduction in the ATP concentration (from 3-5 mm to 30-50 mum) increased the apparent affinity of the Na pump for K(o) (+) about 8-fold. In addition, the maximal rate of K(o) (+)-stimulated Na efflux was reduced by a similar factor. Analogous results were seen in axons dialysed with 310 mm-K(i) (+) or without K(i) (+).4. The relative effectiveness of external monovalent cations as activators of the Na efflux was a function of the ATP concentration inside the axon. With 3-5 mm-ATP the order of effectiveness was K(+) > NH(4) (+) > Rb(+). With 30-50 mum-ATP the sequence was NH(4) (+) >> K(+) >> Rb(+). These results were not affected by the removal of K(i) (+).5. When the ATP concentration was 3 mm and the Na(i) (+) concentration 70 mm, the removal of K(i) (+) produced a slight and reversible increase in the total efflux of Na (15%) and no change in the ATP-dependent Na efflux. When the ATP concentration was reduced to 30-50 mum, or the Na(i) (+) concentration lowered to 5-10 mm, the removal of K(i) (+) reversibly increased the total and the ATP-dependent efflux of Na. The largest increase in Na efflux was seen when both ATP and Na(i) (+) were simultaneously reduced. The ATP-dependent extra Na efflux resulting from the exclusion of K(i) (+) was abolished by 10(-4)m-ouabain in the sea waters.6. The increase in the ATP-dependent Na efflux observed in axons dialysed with 0 K(i) (+) + 10 mm-K(+) ASW was not seen in axons perfused with 310 mm-K(i) (+) + 450 mm-K(+) ASW. However, both experimental conditions gave rise to a similar (and small) ATP-independent and ouabain-insensitive efflux of Na. This indicates that the effects on the Na pump of removing K(i) (+) are not due to the simultaneous membrane depolarization. In addition, it suggests that K(i) (+) has an inhibitory effect on the Na pump, and that that effect is antagonized by Na(i) (+) and ATP.7. The present results are consistent with the idea that the same conformation of the Na pump (and Na,K-ATPase) can be reached by interaction with external K(+) after phosphorylation and with internal K(+) before rephosphorylation. This enzyme conformation produces an enzyme-K complex from which K(+) ions are not easily released unless high concentrations of ATP are present. This also stresses a non-phosphorylating regulatory role of ATP. PMID- 6273537 TI - Transport of cholecystokinin-octapeptide-like immunoreactivity toward the gut in afferent vagal fibres in cat and dog. AB - 1. The distributions of gastrin- and cholecystokinin-like immunoreactivities in the dog and cat vagus nerves have been studied after nerve section and ligation. 2. In dogs, there was an increase in cholecystokinin-octapeptide-like immunoreactive material on the cranial side of ligatures on the thoracic or cervical vagi. When pairs of ligatures were tied on the cervical vagi there was accumulation proximal, and a slight decrease distal to, the upper ligature. There was also a modest increase distal to the lower ligature. 3. In cats, section of the vagus above the nodose ganglion, and hence degeneration of the efferent fibres, did not prevent increases in cholecystokinin-octapeptide-like immunoreactivity on the cranial side of ligatures which were later tied below the ganglion. Removal of the superior cervical ganglion had no effect on the accumulation of immunoreactive material above the ligatures. Section of the vagus below the nodose ganglion, and hence degeneration of both afferent and efferent fibres, abolished the accumulation on the cranial side of ligatures which were later tied below the section. Cholecystokinin-octapeptide-like material is therefore localized to afferent fibres with cell bodies in the nodose ganglion. 4. Immunoreactive forms were characterized by gel filtration and ion exchange chromatography, and the use of region-specific antisera. In all cats, and all but one dog, a molecule with the properties of sulphated cholecystokinin octapeptide was found to predominate. In some cats (30%) and dogs (26%) a molecule with the properties of heptadecapeptide gastrin (G17) was identified; concentrations of G17 were generally low compared with cholecystokinin octapeptide. In three dogs (20%) there was an accumulation of heptadecapeptide gastrin above the ligatures. 5. Axonal transport of cholecystokinin octapeptide in the vagus is consistent with a neuro-regulatory role for this peptide. However, the functional significance of its localization in afferent fibres, and transport towards the periphery, remains to be determined. PMID- 6273536 TI - Are acetylcholine-induced increases in 42K efflux mediated by intracellular cyclic GMP in turtle cardiac pace-maker tissue? AB - 1. 42K efflux has been measured from small strips of turtle sinus venosus tissue in order: (a) to characterize further the pharmacology of the acetylcholine response and (b) to test whether cyclic guanosine 3':5'-monophosphoric acid (cyclic GMP) is the intracellular mediator of this response. 2. The 42K wash-out curves show that the fractional escape rate (FER) of 42K efflux is nearly constant after 60-80 min, indicating that after this time period 42K FER is controlled by barrier-limited diffusion from a single intracellular compartment. 3. The threshold of the dose-response relationship for the acetylcholine-induced increase in 42K FER is about 10(-8) M and the Km is 2.75 x 10(-7) in non eserinized preparations. 4. This acetylcholine response is completely blocked by atropine; but nicotinic blockers produce no detectable reduction of it. 5. Exogenous application of lipid-soluble analogues of cyclic GMP (dibutyryl or 8 bromo-cyclic GMP applied at 2-3 mM for 30 min) failed to mimic the acetylcholine induced augmentation of 42K FER. 6. Experiments in which sodium nitroprusside (5 x 10(-4) M for 30 min) was applied in order to stimulate the guanylate cyclase and hence produce a large, maintained increase in intracellular cyclic GMP failed to show a significant increase in 42K FER. 7. When acetylcholine (10(-6)M) was applied in the presence of O[Ca2+]0 (in an attempt to inhibit the guanylate cyclase) there was no significant reduction in the acetylcholine-induced increases in 42K FER. 8. Hence, these three indirect tests indicate that the muscarinic acetylcholine-induced increase in 42K FER in cardiac pace-maker tissue is unlikely to be mediated entirely by changes in the levels of intracellular cyclic GMP. PMID- 6273538 TI - Pentobarbitone interference with inhibitory synaptic transmission in crayfish stretch receptor neurones. AB - 1. The effect of pentobarbitone (PB) on GABA-ergic inhibition was investigated in the isolated crayfish stretch receptor. The soma of the slowly adapting neurone was impaled with two micro-electrodes to give an accurate determination of membrane conductances. 2. Application of PB in concentrations from 10(-6) to 10( 3) M increased the rise time constant of the inhibitory post-synaptic potential (i.p.s.p.). The i.p.s.p. percentage amplitude and decay time constant were also increased in eight out of twelve neurones. On prolonged exposure, the percentage amplitude declined at a rate dependent upon the dose and the frequency of stimulation until the i.p.s.p. became undetectable. 3. The response to ionophoretically applied GABA remained essentially unaltered in the presence of PB, but the falling phase was prolonged by up to 8% in four of the ten neurones tested. Resting membrane conductance, i.p.s.p. driving force (i.p.s.p. reversal potential minus resting membrane potential), and parameters of the anti- and orthodromic action potential were not significantly affected. 4. Removal of PB after prolonged exposure usually caused an immediate increase in i.p.s.p. percentage amplitude but the i.p.s.p. rising phase remained slowed. 5. Application of excess extracellular GABA only affected the i.p.s.p. percentage amplitude after it had been reduced by PB. It transiently increased the attenuated i.p.s.p. percentage amplitude in the presence of PB, and after the removal of PB permanently increased the amplitude to its original value. 6. Nipecotic acid and cis-1,3-aminocyclohexane carboxylic acid, inhibitors of GABA re-uptake, slightly increased the i.p.s.p. percentage amplitude, and prolonged the falling phase but did not affect the rising phase. The percentage amplitude declined on prolonged exposure. 7. We conclude that PB has no electrophysiologically demonstrable post-synaptic action in the crayfish stretch receptor neurone, but it inhibits the presynaptic release and re-uptake of GABA. PMID- 6273539 TI - Involvement of Na+-K+-ATPase in p-aminohippurate transport by rabbit kidney tissue. AB - 1. The relation between renal accumulation of p-aminohippurate (PAH), Na+-K+ ATPase activity, and the transmembranal Na+ gradient has been investigated by the use of cortex slices of rabbit kidney. 2. A moderate stimulation of PAH uptake rate was observed under anaerobic conditions in the presence of an extracellular to-intracellular directed gradient of Na+. 3. Inhibition of aerobic accumulation of PAH by ouabain was related to a decrease in Na+-K+-ATPase activity, as evidenced by changes in tissue concentrations of Na+ and K+ and by measurements of high-affinity binding of ouabain to the slices. 4. Accumulation of PAH in media with various concentrations of Na+ and K+ resembled the effect of the cations on the ATPase activity of an isolated preparation of Na+-K+-ATPase. 5. Counteraction of the inhibitory effect of ouabain on PAH accumulation by a high concentration of K+ in the medium was related to retention of Na+-K+-ATPase activity, as evidenced by preservation of tissue--medium gradients of Na+ and K+. 6. The present data provide strong evidence for the involvement of Na+-K+-ATPase in the energization of renal PAH accumulation. However, it appears probable that a metabolic (Na+-gradient-independent) component, in addition to a Na+ gradient, is essential for the attainment of high accumulation ratios of PAH by intact renal cells. PMID- 6273540 TI - Effects of catecholamines and cyclic amp on excitation--contraction coupling in isolated skeletal muscle fibres of the frog. AB - 1. In skeletal muscle the presence of a positive inotropic effect induced by adrenaline has been a matter of controversy. If it exists, it could be due to catecholamines acting on the actomyosin system, on the sarcoplasmic reticulum (SR) Ca2+ pump or on the release or influx of Ca2+. We investigated these possibilities by using intact, split and skinned skeletal muscle fibres. We also investigated whether adrenaline acts directly or through cyclic AMP. 2. Catecholamines produced an increase in twitch tension and in maximum rates of tension development and tension decay. The inotropic effect took 3 min to appear and 8 min to reach its maximum level. With tetanic stimulations the extra force appeared only at the beginning of the tetanus while approaching the same maximum level, and tended to disappear faster, the higher the frequency of stimulation. At 4 shocks/sec the peak twitch tension with catecholamines decreased during the first seven to ten twitches and became steady afterwards at a level that was still greater than the control. 3. Resting and action potentials showed no important changes in the presence of adrenaline that could explain the inotropic effect. 4. In split fibres the force produced with the release of Ca2+ from the SR by caffeine was 60-100% larger when cyclic AMP was added to the previous loading solution. In skinned fibres adrenaline given directly to the interior of the cell produced no changes in contraction--relaxation cycles induced by fixed amounts of Ca2+ applied with a pipette. 5. These results strongly suggest that catecholamines through cyclic AMP stimulate the SR Ca2+ pump, increasing thereby the concentration of Ca2+ within the SR. This extra Ca2+ when released during subsequent activation may produce the increase in twitch tension. PMID- 6273541 TI - Neuromuscular transmission and smooth muscle membrane properties in the guinea pig ear artery. AB - Effects of noradrenaline and alpha-adrenoceptor blocking agents on neuromuscular transmission of the guinea-pig ear artery were assessed using the micro-electrode method.1. The mean membrane potential, and length and time constants of the longitudinally oriented muscle cells were -64.5 +/- 5 mV (n = 150), 1.03 +/- 0.16 mm (n = 15) and 410 +/- 40 msec (n = 7) respectively. From the current-voltage relationship, weak outward current pulses produced an anomalous rectification of the membrane while stronger intensities produced a normal rectification of the membrane with a depolarization over 10-15 mV.2. Brief stimulation (0.1-0.5 msec) of the tissue produced an excitatory junction potential (e.j.p.). Facilitation produced by repetitive stimulation was evident only on very rare occasions. Higher stimulus intensities caused a stepwise increase of the amplitude of e.j.p.s.3. Spontaneously generated miniature excitatory junction potentials (m.e.j.p.s) were recorded from the muscle membrane. In many cells, the interval and amplitude histograms of m.e.j.p.s showed skew curves. On rare occasions, a bell-shaped amplitude distribution (quantal release of packeted noradrenaline (NA)) was observed.4. NA (> 3 x 10(-7)m) depolarized the membrane and increased the membrane resistance, as measured from the amplitude of the electrotonic potential. Phentolamine suppressed the NA-induced depolarization. However, high concentrations of phentolamine (> 10(-5)m) depolarized the membrane and increased the membrane resistance.5. NA (10(-8)m) caused no change in membrane potential though it suppressed the amplitude of an e.j.p. produced by a single stimulus, but did not suppress the amplitude of the subsequent e.j.p.s evoked by repetitive stimulation (0.2-2.0 Hz). A higher concentration of NA (3 x 10(-7)m) depolarized the membrane and markedly suppressed the amplitude of e.j.p.s. On the other hand, NA (10(-8) or 2 x 10(-8)m) generated burst discharges of m.e.j.p.s between silent periods or random generation.6. Phentolamine (10(-6)m) markedly enhanced the amplitude of e.j.p.s and caused a smooth facilitation in response to a train of stimuli with no effect on the membrane potential. A stepwise change in the amplitude of e.j.p.s was no longer observed at any given stimulus frequency and intensity.7. Phenoxybenzamine (10(-7)m) suppressed the amplitude of e.j.p.s with no change in the membrane potential.8. The results led to the conclusion that, at a concentration which has no effect on the post-junctional muscle membrane, NA and phentolamine seemed to be more effective on prejunctional adrenoceptors, while phenoxybenzamine seemed to have a greater effect on post-junctional adrenoceptors. NA depresses adrenergic transmission by negative feed-back, while the enhancing action of phentolamine can be explained partly by blocking of prejunctional adrenoceptors and, in addition, by an increase of NA release. PMID- 6273542 TI - Development of sodium, calcium and potassium channels in the cleavage-arrested embryo of an ascidian. AB - 1. The cleavage of the embryos of an ascidian, Halocynthia roretzi, was arrested with cytochalasin B (1 microgram/ml.) at the 16- or 64-cell stage. These cleavage arrested embryos were still able to develop membrane excitability. 2. In the cleavage-arrested 64-cell embryos at the time when control embryos became hatched larvae, Ca spikes were evoked in the presumptive muscle blastomeres, and Na- and Ca-dependent action potentials were induced in some ectodermal blastomeres. 3. Membrane currents of the cleavage-arrested 16-cell embryos were recorded with the voltage-clamp technique and analysed as a function of developmental time at 15 degrees C. For this purpose, intact eggs, 4- and 8-cell embryos were also used. The cleavage-arrested embryos behaved electrically like single cells, due to tight electrical coupling between blastomeres. After the 25-hr stage decoupling occurred. 4. Both Na and Ca currents decreased during the initial 10 hr. Na current became less than one third and Ca current almost disappeared. At 17 hr both Na and Ca currents increased again. 5. The potential-dependence of the Na and Ca currents after 17 hr was similar to that in the egg, although substantial parallel shifts in the current-voltage relations were observed: a 5 mV positive shift for the Na current and a 15-20 mV negative shift for the Ca current. 6. Delayed (outward) K current developed gradually until 20 hr and then increased abruptly. The activation level for the delayed rectification was markedly negative (around -10 mV) in comparison with that of the egg (around + 100 mV). Anomalous (inward) K current, on the other hand, increased gradually without changes in the potential-dependence throughout development. 7. The results suggest that the differentiation of excitable membranes in the ascidian embryo does not involve changes in the properties of the individual channels, but rather changes in the numbers of various kinds of ion channels. PMID- 6273543 TI - Free calcium ions in neurones of Helix aspersa measured with ion-selective micro electrodes. AB - 1. Intracellular free calcium concentration, [Ca2+]i, was measured in giant neurones of the sub-oesophageal ganglia of Helix aspersa, using Ca-selective micro-electrodes containing a PVC-gelled, neutral-ligand sensor. 2. In calibration solutions the electrodes had a virtually ideal, Nernstian, response down to 1 microM-Ca2+ in the presence of 0.125 M-K+, 18-24 mV from 1 to 0.1 microM-Ca2+ and 8-14 mV from 0.1 to 0.01 microM-Ca2+. Interference from H+ and Mg2+ was negligible. The small response to Na+ at sub-micromolar Ca2+ was taken into account, when necessary, in measurement of [Ca2+]i. 3. Measurements of basal [Ca2+]i were made in ganglia from animals kept only a few weeks in captivity, in a bathing solution equilibrated with air and containing 2 mM-Ca2+. In thirteen measurements from impalements which met stringent criteria for electrode performance and cell viability, the mean basal pCa (--log10[Ca2+]) was 6.77 +/- 0.07 (S.E.), corresponding to a mean free Ca2+ concentration of 0.17 microM. 4. The basal [Ca2+]i in neurones from a group of snails kept hibernating for several months was higher, mean pCa 6.15, for ganglia handled in 2 mM-Ca2+ solution. 5. Intracellular injections of Ca2+ or EGTA raised and lowered, respectively, the indicated basal [Ca2+]i, showing that the electrodes responded appropriately inside the cells and that unknown or untested components of cytoplasm were not significantly interfering with the Ca-sensor. 6. Altering the external Ca2+ concentration between 0.1 and 10 mM usually produced only small, +/- 0.1 pCa units, changes in basal [Ca2+]i of satisfactorily impaled, quiescent cells. 7. In cell 1F, which has repetitive spikes with a substantial Ca current, changes in Ca gradient or blockade of voltage-dependent Ca channels sometimes markedly altered [Ca2+]i, showing that Ca entry with the spikes was elevating [Ca2+]i. 8. Replacing external Na+ with Li+ or bis(2-hydroxyethyl)dimethylammonium had little effect on [Ca2+]i. 9. Elevating CO2 to 5% or 79% lowered [Ca2+]i by an average of 0.16 and 0.26 pCa units respectively. PMID- 6273545 TI - Force-velocity relation in deuterium oxide-treated frog single muscle fibres during the rise of tension in an isometric tetanus. AB - 1. The force-velocity (P-V) relation from a single fibres isolated from the semitendinosus muscle of the frog was determined at pre-set times during the rise of tension and the plateau of isometric tetani. The controlled-velocity release method was used. Experiments were performed at about 2.25 micrometers sarcomere length and at 3-4 degrees C or at 19-21 degrees C. 2. Replacing H2O with D2O resulted in a rapid large reduction of the peak twitch tension and of the speed of development of twitch and tetanic tensions. The tetanic tension (P0) was usually reduced, in certain fibres to as low as 5% of the value in H2O-Ringer solution. 3. The depression of twitch and tetanus characteristics was followed by a recovery, the duration of which varied greatly in different fibres. During the recovery period previous conditioning activity potentiated the tetanus characteristics. 4. After the end of the recovery period in D2O-Ringer solution both the peak twitch tension and the speed of development of tetanic tension was still greatly depressed, whereas the value of P0 was slightly greater than in H2O Ringer. The speed of rise of isometric tension after a quick release imposed at the tetanus plateau was reduced in D2O-Ringer, usually to about 50% of the value in H2O-Ringer. 5. D2O increased the development time of the P-V relation and produced a conspicuous increase in the degree of its curvature. The value of V0 (the velocity of shortening at zero load) was not significantly depressed by D2O and it was the same independent both of the time after the beginning of stimulation and of the isometric tension at which the measurement was made. The P V relation attained its final characteristics before the isometric tension reached the plateau. During the recovery period in D2O-Ringer, at the plateau of isometric tetani of different size, the relative force exerted at a given velocity of shortening was constant. 6. In D2O-treated fibres, NO3- and caffeine (i) potentiated the peak twitch tension and the speed of development of tetanic tension without affecting significantly the speed of the redevelopment of tension after a quick release imposed at the tetanus plateau and (ii) reduced the development time of the P-V relation, but did not affect either the degree of its curvature or the value of V0 and P0. 7. The results are discussed by assuming that the release of Ca2+ from the sarcoplasmic reticulum is a rate-limiting process for the development of activation and in turn for the development of isometric tension. In terms of the cross-bridge model of Huxley (1957), the time or Ca2+-dependent factor of activation appears to be the recruitment of actin sites for cross-bridge formation, whereas the value of the rate constants regulating the cross-bridge kinetics appears to be time and Ca2+-independent. PMID- 6273544 TI - Electrophysiology of mammalian inferior olivary neurones in vitro. Different types of voltage-dependent ionic conductances. AB - The electrophysiological properties of guinea-pig inferior olivary (I.O.) cells have been studied in an in vitro brain stem slice preparation. 1. Intracellular recordings from 185 neurones in this nucleus reveal that antidromic, orthodromic or direct stimulation generates action potentials consisting of a fast spike followed by an after-depolarizing potential (ADP). The ADP had an amplitude of 49 +/- 8 mV (mean +/- S.D.) and a duration which varied over a wide range with the level of depolarization. This ADP is followed by an after-hyperpolarizing potential (AHP) having an amplitude of 12 +/- 3 mV (mean +/- S.D.) from rest and lasting up to 250 msec. The AHP shows a rebound depolarization wave. 2. Synaptic activation may be obtained by peri-olivary stimulation with a bipolar electrode located in the immediate vicinity of the I.O. nucleus. These potentials are a mixture of depolarizing and hyperpolarizing synaptic events which can be reversed by direct membrane polarization. 3. Addition of tetrodotoxin (TTX) to the bath, or removal of extracellular Na, abolishes the fast initial action potential but does not modify the ADP or the AHP. Blockage of Ca conductance by Co, Mn, Cd or D600, or replacement of Ca by Mg, abolishes the ADP--AHP sequence. 4. Hyperpolarization of the neurone uncovers a low-threshold Ca conductance which is inactivated at rest and has similar pharmacological properties to the ADP. This low-threshold spike plays a central role in the rebound potential following the AHP. 5. Simultaneous impalement of I.O. neurone pairs demonstrated the presence of electrotonic coupling between neurones, which is especially prominent in the medial accessory olive. PMID- 6273546 TI - The sodium current underlying the responses of toad rods to light. AB - 1. Intracellular responses were recorded from single rods in the retina of the toads Bufo bufo and Bufo marinus during exposure to solutions in which sodium was replaced by equimolar amounts of choline. 2. Upon moderate reduction (80 and 50 mM) of the external sodium the size of responses to bright flashes decreased as a consequence of both an increase in the resting potential and a decrease of the membrane potential at the peak, while the level of the plateau remained fairly constant. 3. Upon reduction of the external sodium to 22 mM or less, rods hyperpolarized to about the plateau level and failed to respond to illumination. Under these circumstances, membrane depolarization induced by an increased external potassium did not restore the cell responsiveness. Addition of 2-5 mM caesium hyperpolarized the membrane and partially restored the photoresponse. 4. Complete replacements of external sodium with potassium depolarized the rod by 40 +/- 10 mV, and no voltage responses to light could be detected. 5. In the presence of caesium, a nearly complete blockage of the photoresponses was obtained when the external sodium was 5 mM or less. Further reductions of the external sodium did not invert the photoresponses. Application of caesium when the external sodium was nominally zero induced a transient hyperpolarization followed by a slow decay. 6. During exposure to steady illumination, the dependence of the plateau level on the external sodium slowly increased. 7. These results indicate that the ionic current which is directly modulated by the light depends primarily on the external sodium. They suggest also that the current associated with the voltage- and time-dependent process responsible for the sag from peak to plateau of the response to a bright flash of light may have multiple components. PMID- 6273547 TI - Adrenergic transmissions in the guinea-pig mesenteric artery and their cholinergic modulations. AB - 1. Neuromuscular transmission in the guinea-pig mesenteric artery and the effect of acetylcholine (ACh) on transmission were investigated by the micro-electrode method. 2. The membrane potential, length and time constant of the smooth muscle of the mesenteric artery were found to be -69.6 +/- 1.9 mV, 0.81 +/- 0.15 mm and 129 +/- 42 msec, respectively. 3. Perivascular nerve stimulation produced an excitatory junction potential (e.j.p.) and repetitive stimulation produced facilitation. When the amplitude of an e.j.p. reached threshold, a spike was evoked. 4. Very rarely, miniature e.j.p.s. were recorded. The amplitude histogram showed a skew distribution. Increases in the stimulus intensity enlarged the amplitude of e.j.p.s. as a stepwise manner. These results indicate multiple innervation to the muscle cells. 5. The time constant of the falling phase of an e.j.p. was consistently larger than that of the muscle membrane. 6. ACh, in concentrations of less than 10(-8) M, suppressed the e.j.p. amplitude without change in the membrane potential and resistance of the muscle membrane, but accelerated facilitation. These ACh actions were suppressed by pre-treatment with atropine. 7. ACh suppressed neither the conduction velocity of excitation of adrenergic nerves nor the number of nerves contributing to the generation of an e.j.p. 8. These results suggest that pre- and post-junctional muscarinic receptors possess different sensitivities to ACh, and a low concentration of ACh (less than 10(-8)M solely suppressed the release of noradrenaline by activating the pre-junctional muscarinic receptors. PMID- 6273549 TI - Pre- and post-synaptic actions of botulinum toxin at the rat neuromuscular junction. AB - 1. Extensor digitorum longus muscles of rats were paralysed with local, non lethal doses of botulinum toxin Type A (BoTx). At 2 and 7 days after toxin injection, the nerve-muscle preparations were excised and end-plate currents analysed at 23 degrees C by dual-micro-electrode voltage clamp. 2. At 2 days after BoTx injection, the growth time of miniature end-plate currents (m.e.p.c.s) increased from a rather narrow range with a mean of 0.59 to a mean of 1.35 ms with a large variability between m.e.p.c.s. End-plate currents (e.p.c.s) were reduced compared to unpoisoned muscle. The decay phase of m.e.p.c.s and e.p.c.s, the growth phase of e.p.c.s and the voltage sensitivity of m.e.p.c.s were unchanged. 3. At 7 days after BoTx injection, the findings were similar to 2 days except that the time constant of the decay phase of m.e.p.c.s. and e.p.c.s. was about twice a long as normal and that the voltage sensitivity of m.e.p.c.s was increased. 4. The acetylcholine null potential (about 0 mV) was unchanged after treatment with BoTx. 5. The increase in the growth time of m.e.p.c.s compared to e.p.c.s following the injection of BoTx suggests that the poisoning, besides blocking quantal release, affects the time course of spontaneous but not that of evoked release. After BoTx poisoning the trans-synaptic diffusion of a majority of spontaneously released transmitter quanta seems to occur more slowly or from areas more distant from the highest concentration of the post-synaptic receptor than that of evoked release. 6. The increase in the decay phase of m.e.p.c.s and e.p.c.s and its increased voltage sensitivity observed in muscles poisoned for 7 days with BoTx suggest that appearance at the end-plate of a population of new receptors with a prolonged ion channel opening time similar to that previously described for extrajunctional receptors after denervation and for junctional receptors during development. PMID- 6273548 TI - Transient, axotomy-induced changes in the membrane properties of crayfish central neurones. AB - 1. In crayfish, the normally passive, non-spiking somata of certain unipolar, efferent neurones became spiking within 36 hr of axotomy. 2. The changes persisted for approximately 2 weeks and then waned. The decline in excitability occurred independently of regeneration, and excitability was not restored by recutting the axon stump. 3. The neuropilar processes also became capable of supporting spikes, but synaptic transmission onto the cells and the spike threshold for orthodromic activation were unchanged, as was the gross structure of the neurone. 4. In somata which normally spike, electrogenicity was nevertheless increased, as evidenced by soma spikes that were larger, faster rising, and easier to evoke. 5. We tested for post-axotomy excitability changes in a variety of identified neurones. Every type (n = 5) of phasically active efferent we tested responded as above, as did all three phasic interneurones. One class of spontaneously active interneurones and one spontaneously active efferent did not respond to axotomy. 6. Extensive damage to afferents did not initiate changes in efferents of the same ganglion, nor did it interfere with changes induced by axotomy of the efferents. 7. Transection of the larger of the two main branches of the phasic flexor inhibitor induced soma excitability, but cutting the smaller branch did not. However, after the excitability caused by cutting the larger branch waned, transection of the smaller branch then induced excitability. 8. Neurones with longer axon stumps took longer to develop soma excitability. PMID- 6273551 TI - A comparison of the treatment of otitis externa with 'Otosporin' and aluminium acetate: a report from a services practice in Cyprus. AB - We studied 126 cases of otitis externa among the population of a British forces base in Cyprus during one season. The causes of infection, the infective agents involved and the treatment of the observed cases in general practice are discussed. The patients were treated with either antibiotic/steroid ear drops (;Otosporin') or with aluminium acetate ear drops. We found no significant difference between the two. PMID- 6273550 TI - Effects of quinine and apamin on the calcium-dependent potassium permeability of mammalian hepatocytes and red cells. AB - 1. K-sensitive electrodes placed in the extracellular fluid have been used to show that ATP and noradrenaline cause a rapid loss of up to 10% of the K content of isolated guinea-pig hepatocytes. 2. The hypothesis tha this response is a consequence of a rise in the K permeability of the hepatocyte membrane triggered by an increase in cytosolic Ca is supported by the finding that the divalent cation ionophore A23187 also initiated K loss, in this instance of up to 20-25% of the amount in the cells. 3. Under similar conditions A23187 caused a transient increase, followed by a larger decrease, in the 45Ca content of guinea-pig hepatocytes equilibrated with this isotope. The decrease alone was seen with ATP and noradrenaline. 4. Quinine (1 mM) and the bee venom neurotoxin apamin (10 nM) greatly reduced the effect of ATP, noradrenaline and A23187 on K content without affecting the changes in 45Ca movement. 5. Apamin (10 nM) also abolished the increase in 42K efflux which follows the application of the alpha-adrenoceptor agonist amidephrine to rabbit liver slices; the concurrent rises in 45Ca efflux and glucose release were unaffected. 6. It was concluded that quinine and apamin are able to block either the Ca-dependent K channels present in guinea-pig and rabbit liver cell membranes or the mechanism that controls them. 7. Surprisingly, rat hepatocytes took up rather than lost K when treated with the concentrations of ATP, noradrenaline or A23187 that initiated K loss from guinea-pig cells. This response was greatly reduced by ouabain. 8. Application of large concentrations of A23187 to rat hepatocytes caused K loss associated with cell death. 9. The influence of apamin (10-1000 nM) and quinine (200-1000 micro M) on the Ca dependent K permeability of red blood cells and ghosts was also studied. Apamin was without effect even when applied to both sides of the ghost membrane, whereas quinine caused inhibition, as reported by others. 10. The results suggest that Ca dependent K channels or carriers are present in the membranes of liver cells of the guinea-pig and rabbit, but are either lacking or inactive in rat liver. The finding that apamin blocks this mechanism in hepatocytes but not in erythrocytes may mean that the channels differ in these cells. PMID- 6273552 TI - Soft tissue uptake of bone seeking radionuclide in amyloidosis. AB - The diagnosis of amyloidosis is often difficult due to variable clinical manifestations and the need for biopsy confirmation. We evaluated the use of bone scanning radionuclides in an effort to delineate soft tissue amyloidosis. Three of 7 patients with proven amyloid reviewed retrospectively, and 3 of 17 studied prospectively had abnormal soft tissue uptake of radionuclide, most commonly diffuse hepatic uptake. Amyloidosis should be considered in any patient with diffuse hepatic uptake or other soft tissue uptake of bone seeking radionuclides. A mechanism for the abnormal uptake (increased calcium content of the involved tissue) is postulated based on in vitro tissue analysis for calcium. PMID- 6273553 TI - Calcium pyrophosphate dihydrate deposition disease and familial hypomagnesemia. AB - A 40-year-old woman presented with calcium pyrophosphate synovitis and chondrocalcinosis. She was subsequently found to have hypomagnesemia, as did her 22-year-old son. Metabolic studies demonstrated normal gastrointestinal absorption of magnesium, and impaired renal conservation of magnesium without other evidence of renal tubular dysfunction. It seems likely that a genetically determined abnormality of magnesium metabolism was responsible for the occurrence of chondrocalcinosis in this patient. PMID- 6273555 TI - [The effect of cardiac glycosides and electrolytes on cell membrane sodium, potassium-adenosine triphosphatase]. AB - A brief summary is given of the structure of the cell membrane and the enzyme system which actively transports Na+ and K+ ions across it. The function of different ions in this process is discussed as well as the effect of cardiac glycosides on ion transport. The utilization of electrolytes to combat cardiac glycoside poisoning and an indirect method for demonstrating the presence of cardiac glycosides in the organs of ruminants, suspected of being poisoned by these toxins, is described. PMID- 6273554 TI - The effect of calcium and magnesium ions on calcium pyrophosphate crystal formation in aqueous solutions. AB - Calcium pyrophosphate crystal formation has been associated clinically with hypercalcemic states (hyperparathyroidism) and hypomagnesemia. We studied aqueous solutions at pH 7.4, 37 degrees C, [Na+] = 140 nM over a range of calcium chloride/magnesium chloride/sodium pyrophosphate concentrations to determine the effect of calcium and magnesium ions on crystal formation. We found that CPPD(T) and CPPD(M) could form under different ionic conditions. Low [Mg++] and [PPi] favoured CPPD(T) whereas higher [Mg++] and [PPi] favoured CPPD(M). At [Mg++] = 1.0 mM a calcium magnesium pyrophosphate crystal phase designated CMPP2 formed. As [Mg++] affects the crystal phase formed more than equimolar [Ca++], we conclude that ionic magnesium deficiency may be a clinically important determinant in calcium pyrophosphate dihydrate crystal formation. PMID- 6273556 TI - Infection of the central nervous system of horses with equine herpesvirus serotype 1. AB - During the last 2 years different equine herpesviruses serotype 1 strains have been isolated from cases of paretic or paralytic disease among horses in the Federal Republic of Germany. In this paper the available information is collated and briefly reviewed. A short description of the symptoms and the possible mechanism of the pathogenesis are given. PMID- 6273557 TI - Feline leukaemia virus infection. AB - All aspects, including aetiology, incidence, transmission, immunity, clinical signs, pathogenesis, pathology and diagnosis of feline leukaemia virus infection in domestic cats are considered with a bias towards the more clinical aspects for the benefit of private practitioners. Non-neoplastic disease and possible methods of control are also discussed. PMID- 6273558 TI - Mesoionic purinone analogues as inhibitors of cyclic-AMP phosphodiesterase: a comparison of several ring systems. AB - Several simple alkyl and aralkyl derivatives of mesoionic thiazolopyrimidines (1) and mesoionic 1,3,4-thiadiazolopyrimidines (2) were found to possess theophylline like activity as inhibitors of cyclic-AMP phosphodiesterase (PDE). Reduction of the C2-C3 double bond of 1 or replacement of the sulfur atom of 1 or 2 with an N methyl group nearly abolishes activity. Optimal activity appears to be associated with a hydrophobic substituent at the N8 position. The five-membered ring of 1 can be replaced by a pyridine or isoquinoline nucleus without untoward effects. Preliminary kinetic data suggest that the type of enzyme inhibition produced by the mesoionic derivatives is similar to that observed for theophylline. Thus, several novel mesoionic ring systems display activity as inhibitors of cyclic-AMP PDE and can serve as lead compounds for further investigation. PMID- 6273559 TI - 2-l-Rhamnopyranosyl[1,2,4]triazolo[1,5-a]pyridine. 4' and 3' Oxidation products. Synthesis and structure-activity relationships. AB - A series of 2-alpha-L-rhamnopyranosylnitro[1,2,4]triazolo[1,5-a] pyridine C nucleosides was synthesized from the condensation oa thioiminoether with nitro-2 pyridylhydrazines. Catalytic reduction afforded the corresponding amino derivative. A 1',2' unsaturated C-nucleoside was also obtained by two different routes. Selective oxidation gave the 3'- and 4'-ketonucleosides. The cytotoxic properties of the nucleosides, as well as their effect on viral transformation and replication, were described. The nitro derivatives inhibit viral replication, but at toxic doses; the introduction of a keto function leads to a product which inhibits the replication of murine leukemia virus (MuLV) at noncytotoxic concentrations. The amino derivatives have no significant antiviral effect. PMID- 6273560 TI - Preparation and analgesic properties of amino acid derivatives of (-)-5,9 alpha diethyl-2'-hydroxybenzomorphan. AB - The N-arginyl derivative of methionine-enkephalin (fragment 60-65 of beta lipotropin) has been shown to be equiactive with the parent pentapeptide, despite the fact that the tyrosine amino group in this compound has been neutralized by the formation of an amide linkage. A series of N-(amino acid) derivatives of (-) 5,9 alpha-diethyl-2'-hydroxybenzomorphan was prepared and evaluated for analgesic activity. In vitro activities were found to vary greatly, depending on the nature of the amino acid used. The N-arginyl derivative was found to be equipotent to ( )-5,9 alpha-diethyl-2'hydroxybenzomorphan and also to methionine-enkephaline in the naloxone binding assay. PMID- 6273561 TI - Purine acyclic nucleosides. Nitrogen isosteres of 9-[(2 hydroxyethoxy)methyl]guanine as candidate antivirals. AB - A number of nitrogen analogues of 9-[(2-hydroxyethoxy)methyl]guanine [acylovir, Zovirax] containing amine functions in the side chain were synthesized and tested for antiviral activity. These purine acyclic nucleosides were prepared by reaction of tris(trimethylsilyl)guanine or 2,6-diaminopurine sodium salt with the chloromethyl ethers prepared from N-(2-hydroxyethyl)phthalimide, N-[2-(2 hydroxyethoxy)ethyl]phthalimide, or N-(2-hydroxyethyl)oxazolidin-2-one to give the N-blocked intermediates 5-8. Deprotection with hydrazine or by alkaline hydrolysis gave 9-[(2-aminoethoxy)methyl]guanine (9), 9-[(2-aminoethoxy)methyl] 2,6-diaminopurine (10), 9-[2(2-aminoethoxy]ethoxymethyl]guanine (11), and 9-[[2 [(2-hydroxyethyl)amino]ethoxy]methyl]guanine (12). When tested against herpes simplex virus type 1, only 9 was active with an IC50 = 8 microM. Little or no activity was observed against a range of other DNA and RNA viruses. PMID- 6273562 TI - Synthesis and beta-lactamase inhibitory properties of 2 beta-(chloromethyl)-2 alpha-methylpenam-3 alpha-carboxylic acid 1,1-dioxide. AB - Potassium 2 beta-(chloromethyl)-2 alpha-methylpenam-3 alpha-carboxylate 1,1 dioxide (BL-P2013) and its pivaloyloxymethyl ester were prepared by the conversion of 6-aminopenicillanic acid to p-nitrobenzyl 6 alpha-bromo-2,2 dimethylpenam-3 alpha-carboxylate 1-oxide, which was rearranged with benzoyl chloride and quinoline to p-nitrobenzyl 6 alpha-bromo-2 beta-(chloromethyl)-2 alpha-methylpenam-3 alpha-carboxylate in 65% yield. Oxidation and catalytic hydrogenation afforded BL-P2013, which was found to be a potent inhibitor of various bacterial beta-lactamases and has been found to protect amoxicillin from beta-lactamases in both in vitro and in vivo systems. PMID- 6273563 TI - An R plasmid of broad host-range, coding for resistance to nine antimicrobial agents endemic in Gram-negative nosocomial isolates. AB - Of 3952 clinical isolates of Enterobacteriaceae, 246 exhibited resistance to at least carbenicillin, gentamicin and tobramycin. All these isolates, representing eight genera, were resistant to at least nine antimicrobial agents in common, including the three key antibiotics and streptomycin, kanamycin, sisomycin, ampicillin, cephalothin and sulphonamide. The strains could be subdivided into seven groups depending upon additional resistance traits and some were resistant to as many as 15 antibiotics. When mated with a standard strain of Escherichia coli, 85% of 123 randomly selected donors transferred resistance to at least the nine core antibiotics. Some donors occasionally transferred resistance to two additional antibiotics, neomycin and tetracycline, while one Citrobacter freundi donor always transferred linked resistance to all 11 drugs. Although many donors were found to harbour more than one species of plasmid DNA, all except a strain of C. freundi contained at least a plasmid of mol. wt 89 x 10(6). Analysis of E. coli transconjugants showed this plasmid to be responsible for transferable resistance to the nine core antibiotics. Restriction-endonuclease analysis indicates that the 89 x 10(6) plasmids originating from different isolates were essentially identical with each other. These results show that a particular R plasmid has established itself among the Enterobacteriaceae at Hines VA Hospital. This R plasmid appears to be the predominant genetic element responsible for linked resistance to carbenicillin, gentamicin and tobramycin among these hospital-associated bacteria. PMID- 6273564 TI - Rapidity of antigenic modulation of Bordetella pertussis in modified Hornibrook medium. AB - Modulation of Bordetella pertussis was induced by growth in Hornibrook medium with a high content of magnesium sulphate (C-medium); four pathophysiological activities in the cells or in the whole culture were measured at intervals. Modulation, shown by the extensive loss of protective antigen, histamine sensitising factor, leuko-cytosis-promoting factor, heat-labile toxin and X-mode specific envelope proteins, occurred during the first 10 h of incubation of X mode cells in C-medium at 35 degrees C and before the onset of the logarithmic phase of growth. The rapidity of these losses was greater than could be explained by a simple growth-dilution effect and did not appear to be due to release of activity into the culture fluid. It seemed, therefore, that there was selective destruction of pathophysiological activities as well as cessation of synthesis. The activities appeared to be lost at different rates. Mouse-protective activity was lost more rapidly than histamine-sensitising and leukocytosis-promoting activities; heat-labile toxicity was lost more slowly or less completely. Modulation was shown to be easily reversed. PMID- 6273566 TI - Cell junction and cyclic AMP: 1. Upregulation of junctional membrane permeability and junctional membrane particles by administration of cyclic nucleotide or phosphodiesterase inhibitor. AB - Mammalian cells in culture were exposed to cyclic AMP, dibutyryl cyclic AMP, the phosphodiesterase inhibitor caffeine, or a combination of the last two, while junctional molecular transfer was probed with the series of microinjected, fluorescent-labelled linear molecules Glu, Glu-Glu, Glu-Glu-Glu, and Leu-Leu-Leu Glu-Glu. The junctional permeability for these molecules increased with each of the agents, most markedly with the dibutyryl cyclic AMP-caffeine combination, as the intracellular cyclic nucleotide concentration rose. The junctional permeability effect developed over several hours. When probed with molecules close to the limit of cell-to-cell channel permeation (the most sensitive setting), the effect was detectable both, as an increase in the (relative) junctional transit rate and as an increase in the number of transferring cell interfaces in the test populations. The number of transferring cell interfaces reached a maximum by 4 hr, when the junctional transit rate, hence the junctional permeability, was still rising. Nonjunctional membrane permeability for the probe molecules, as determined by intracellular fluorescence loss, was not significantly changed (nor was there significant nonjunctional cell-to-cell transfer of molecules before or after the treatments). The rise in junctional permeability was associated with an increase in the number of gap junctional membrane particles, as determined by freeze-fracture electron microscopy: the average size of the particle clusters increased, and the frequency of the clusters increased, particularly that of the smaller (and presumably newer) clusters. This effect was blocked by treatments with the protein synthesis inhibitors cycloheximide or puromycin. These agents caused particle diminution (diminution of cluster frequency but not of average cluster size), with or without cyclic nucleotide. The junctional effects may represent a cyclic AMP promoted proliferation of cell-to-cell channels. Some physiological implications, in particular, implications for hormone-regulated tissues, are discussed. PMID- 6273567 TI - Cell junction and cyclic AMP: II. Modulations of junctional membrane permeability, dependent on serum and cell density. AB - Junctional molecular transfer (as indexed by the number of cell interfaces transferring fluorescent-labelled molecules) and concentration of endogenous cAMP were determined in mammalian cells in culture at varying serum concentration and cell density. In several cell types, on stepping the serum concentration from 10% (the concentration to which the cells had been adapted) to zero, the junctional transfer rose (reversibly) within 48 hr, as the endogenous cAMP concentration rose. The junctional transfer was inversely related to serum concentration over a range, most steeply so the transfer of large and charged molecules. One cell type showed no junctional change in response to serum; it showed also no endogenous cAMP change. Junctional transfer varied inversely with cell density over the range of 0.7-7 (10(4) cells/cm2) in 3T3 cells. In cultures seeded to various densities, or growing to various densities on their own, junctional transfer fell with rising density, and so did the endogenous cAMP concentration. Upon downstep from high density, junctional transfer was independent of cell density over the aforementioned range, and so was the endogenous cAMP concentration. These results, in conjunction with the effects of exogenous cAMP described in the preceding paper of this series, point to a cAMP-mediated junctional effect; a possible teleonomy for control of membrane junction is discussed. PMID- 6273565 TI - Density-dependent regulation of cell growth: an example of a cell-cell recognition phenomenon. AB - Cell-to-cell contact can result in a variety of changes in the cell's physiology. For different cell types, this may include both the initiation as well as the cessation of cell growth and changes in the state of differentiation. This review examines in detail one such phenomenon, density-dependent inhibition of growth, which is observed with many fibroblasts in culture. Data are summarized which demonstrate that the cessation of growth at high cell density is in part a consequence of cell-to-cell contact. An approach to the study of the molecular basis of this phenomenon is presented based on the demonstration that plasma membranes, when bound to sparse growing cells, mimic contact inhibition of growth. The present status of attempts to purify plasma membrane proteins responsible for this effect are summarized, and the properties of these membrane proteins are compared to those of previously described "soluble" proteins that inhibit cellular growth. PMID- 6273568 TI - Cell junction and cycle AMP: III. Promotion of junctional membrane permeability and junctional membrane particles in a junction-deficient cell type. AB - The cyclic nucleotide effect on junction was studied in C1-1D cells, a mouse cancer cell type that fails to make permeable junctions in ordinary confluent culture. Upon administration of cyclic AMP, dibutyryl cyclic AMP, dibutyryl cyclic AMP plus caffeine (db-cAMP-caffeine), or cholera toxin (an adenylate cyclase activator), the cells acquired permeable junctions; they became electrically coupled and transferred fluorescent tracer molecules among each other - a transfer exhibiting the molecular size limit of permeation of normal cell-to-cell channels. The effect took several hours to develop. With the db-cAMP caffeine treatment, junctional permeability emerged within two hours in one-fifth of the cell population, and within the next few hours in the entire population. This development was not prevented by the cytokinesis inhibitor cytochalasin B. Permeable junctions formed also in two other conditions where the cell-endogenous cyclic AMP level may be expected to increase: serum starvation and low cell density. After three weeks of starving, the cells of serum, a junctional permeability arose in confluent cultures, which on feeding with serum disappeared within two to three days. At low cell density, namely below confluency, the cells made permeable junctions, unstarved. In cultures of rather uniform density, the frequency of permeable junctions was inversely related to the average density, over the subconfluent range; at densities of about 1 X 10(4) cells/cm2, where the cells had few mutual contacts, 80% of the pairs presumed to be in contact were electrically coupled. In cultures with adjoining territories of high (confluent) and low cell density, there was coupling only in the last, and in this low-density state the cells were also capable of coupling with other mammalian cell types (mouse 3T3-BalbC and human Lesch-Nyhan cells). PMID- 6273569 TI - Partial purification and characterization of (Na+ + K+)-ATPase from garfish olfactory nerve axon plasma membrane. AB - The (Na+ + K+)-ATPase of garfish olfactory nerve axon plasma membrane was purified about sixfold by treatment of the membrane with sodium dodecyl sulfate followed by sucrose density gradient centrifugation. The estimated molecular weights of the two major polypeptide components of the enzyme preparation on sodium dodecyl sulfate gels were 110,000 and 42,000 daltons, which were different from those of the corresponding peptides of rabbit kidney (Na+ + K+)-ATPase. No carbohydrate was detected in the 42,000-dalton component either by the periodic acid-Schiff reagent or by the more sensitive concanavalin A-peroxidase staining procedure. The molecular properties of the garfish (Na+ + K+)-ATPase, such as the Km for ATP, pH optimum, energies of activation, Na and K ion dependence and vanadium inhibition, were, however, similar to those of the kidney enzyme. The partially purified garfish (Na+ + K+)-ATPase was reconstituted into phospholipid vesicles by a freeze-thaw-sonication procedure. The reconstituted enzyme was found to catalyze a time and ATP dependent 22Na+ transport. The ratio of 22Na+ pumped to ATP hydrolyzed was about 1; under the same reconstitution and assay conditions, eel electroplax (Na+ + K+)-ATPase, however, gave a 22Na+ pumped to ATP hydrolyzed ratio of nearly 3. PMID- 6273570 TI - Cyclic AMP and intracellular ionic activities in Necturus gallbladder. AB - Open-tip and liquid ion-exchanger microelectrodes were used to study the effects of cAMP (6mM, added to the serosal medium) on apical membrane potential (Em) and intracellular sodium, potassium, and chloride activities (alphaiNa, alphaiK, alphaiCl) in Necturus gallbladder under open-circuit conditions. Transepithelial potential difference (ETr) was also measured. In the presence of cAMP, alphaiCl fell from about 1.5 times its equilibrium value to a level that corresponded to electrochemical equilibrium across the apical and basolateral cell membranes. Under these conditions alphaiNa decreased and alphaiK increased, Em was unchanged and ETr increased from virtually zero to a small but significant serosal positive value. The cAMP-induced increase in alphaiK was abolished when Cl- -free incubation media were used. Addition of the Ca++-ionophore A23187 (0.5 microgram/ml) to the serosal medium had no effect on Em, ETr, or alphaiCl. When A23187 was added to the mucosal medium, Em and the basolateral membrane potential hyperpolarized by about 20 mV and an increase in the outwardly directed electrochemical driving force for Cl- was observed. These results indicate that cAMP inhibits coupled transapical Na-Cl entry into epithelial cells of Necturus gallbladder and suggest that this inhibition may not be mediated by an increase in intracellular Ca++ concentration. PMID- 6273571 TI - Sulfatide role in the sodium pump. AB - Sodium efflux was studied in 22Na-loaded red blood cells in the presence of arylsulfatase, an enzyme that specifically hydrolyzes sulfatide. Sodium efflux was inhibited in proportion to the amount of arylsulfatase present. Maximum inhibition was almost as high as the efflux obtained in medium with K+ absent. At maximum inhibition 83.2% of the sulfatide content of the fragmented red blood cell membranes was hydrolyzed and ouabain-sensitive (Na+ + K+)-ATPase activity was inhibited by 100%. Sodium efflux, sulfatide content, and (Na+ + K+)-ATPase activity were unaffected with arylsulfatase in the presence of a high concentration of sulfatide. These results indicate that sulfatide plays a specific role in sodium and potassium ion transport. They also suggest that most sulfatide is localized externally in the red blood cell membrane. PMID- 6273572 TI - Protein interactions with lipid bilayers: the channels of kidney plasma membrane proteolipids. AB - Proteolipids extracted from bovine kidney plasma membrane induce irreversible changes in the electrical properties of lipid bilayers formed from diphytanoyl phosphatidylcholine. The interaction with the proteolipid produces channels which are cation selective. At low protein concentrations (i.e., less than 0.6 microgram/ml), the single-channel conductance is approximately 10 pS in 100 mM KCl and 3 pS in 100 mM NaCl. In the presence of protein concentrations above 1 microgram/ml, another population of channels appears. These channels have a conductance of about 100 pS in 100 mM KCl and 30 pS in 100 mM NaCl. Further, these channels are voltage dependent in KCl, closing when the voltage is clamped at values greater than or equal to 30 mV. The steady-state membrane conductance, measured at low voltages, was found to increase proportional to a high power (2 3) of the proteolipid concentration present in one of the aqueous phases. In 100 mM NaCl, the conductance increases at protein concentrations above 5 microgram/ml, whereas in 100 mM KCl it increases at protein concentrations above 0.6 microgram/ml. These measurements indicate that the higher steady-state conductance observed in KCl at a given proteolipid concentration in a multi channel membrane presumably results because more channel incorporate in the presence of KCl than in the presence of NaCl. The two major fractions which comprise the proteolipid complex were also tested on bilayers. It was found that both fractions are required to produce the effects described. PMID- 6273573 TI - K-current fluctuations in inward-rectifying channels of frog skeletal muscle. AB - K currents and K-current fluctuations were recorded in inwardly rectifying K channels of frog skeletal muscle under voltage-clamp conditions. External application of 0.2 to 10 mM Cs reduces the inward mean K current but produces a distinct increase of the spectral density of K-current fluctuations. The additional fluctuations arise from the random blocking by Cs ions. From the variance of current fluctuations, the steady-state current and the probability of the open unblocked channel an effective single-channel conductance gamma* was calculated. Gamma* strongly depends on the external Cs concentration (7.8 pS at 0.2 mM Cs, 2.1 pS at 10 mM Cs). This dependence is interpreted in terms of a two step blocking process: (1) a fast exchange of Cs ions between the external solution and a first binding site inside the channel which leads to the Cs modulated effective single-channel conductance, and (2) a slow Cs binding to a second site deeper in the channel which produces the observed current fluctuations. With this hypothesis we obtained a real single-channel conductance of gamma approximately equal to 10 pS and a real density of n approximately equal to 4 inwardly rectifying channels per micrometer2 of muscle surface area. PMID- 6273575 TI - Bilateral synchronous primary lung cancer. PMID- 6273574 TI - Noise analysis of the K+ current through the apical membrane of Necturus gallbladder. AB - Current noise power spectra of the voltage-channel (V = 0) Necturus gallbladder, exposed to NaCl-Ringer's on both sides contained a relaxation noise component, which overlapped with a 1/f alpha noise component, with alpha being about 2. Substitution of all Na+ by K+ on either the serosal or mucosal side increased the relaxation as well as the 1/f alpha noise component considerably. In Necturus gallbladder both noise components are reduced by addition of 10 mM, 2,4,6 triaminopyrimidine (TAP) or 5 mM BA2+ to the mucosal side, as well as by acidification of the mucosal solution to pH 5 and lower. Five mM of tetraethylammonium (TEA+) added to the mucosal solution, abolished K+ relaxation noise and decreased the 1/f alpha noise component. Applying a Cs+ concentration gradient across the epithelium did not yield relaxation noise. However, if Rb+ was substituted for all Na+ on one side, a Lorentzian noise component appeared in the spectrum. Its plateau was smaller than with KCl-Ringer's on the respective side. These data confirm the existence of fluctuating K+ channels in the apical membrane of the Necturus gallbladder. Furthermore it can be concluded that these channels have the permeability sequence K+ greater than Rb+ greater than Cs+. The inhibition of the fluctuations by mucosal acidification indicates the existence of acidic sites in the channel. The single-channel conductance was estimated to be between 6.5 and 40 pS. PMID- 6273576 TI - Serine insertion caused by the ribosomal suppressor SUP46 in yeast. PMID- 6273577 TI - Identification of the uvrA gene product. PMID- 6273578 TI - Identification of the uvrB gene product. PMID- 6273579 TI - Transposition of the gene cluster CYC1-OSM1-RAD7 in yeast. PMID- 6273580 TI - Oligomeric structure of a simian virus 40 T antigen in free form and bound to DNA. PMID- 6273582 TI - Eukaryotic dinucleotide preference rules and their implications for degenerate codon usage. PMID- 6273581 TI - Kinetic analysis of template-instructed and de novo RNA synthesis by Q beta replicase. PMID- 6273583 TI - Three-dimensional structure of ubiquinol:cytochrome c reductase from Neurospora mitochondria determined by electron microscopy of membrane crystals. PMID- 6273584 TI - Evolution of the beta-globin gene cluster in man and the primates. PMID- 6273586 TI - Mutational variants of the Neurospora crassa NADP-specific glutamate dehydrogenase altered in a conformational equilibrium. PMID- 6273585 TI - rII cistrons of bacteriophage T4. DNA sequence around the intercistronic divide and positions of genetic landmarks. PMID- 6273587 TI - Measurement of the fraction of myosin heads bound to actin in rabbit skeletal myofibrils in rigor. PMID- 6273588 TI - Pf1 filamentous bacterial virus. X-ray fibre diffraction analysis of two heavy atom derivatives. PMID- 6273589 TI - A high-resolution electron microscopy study of nucleosomes from simian virus 40 chromatin. PMID- 6273590 TI - Sequence-dependent variation in the conformation of DNA. PMID- 6273591 TI - Structure of a B-DNA dodecamer. II. Influence of base sequence on helix structure. PMID- 6273592 TI - Calcium entry in the calcium paradox. PMID- 6273593 TI - The dynamic state of liver gap junctions. AB - By the use of a simple, rapid method for the isolation of gap junctions from small amounts of rat liver (2-3 g), we have followed the incorporation of the radiolabeled amino acid precursors 3H-leucine and 35S-methionine into the gap junction protein. In timed studies with 35S-methionine as precursor, the specific activity in the protein is maximal by 4 h after a single injection of 300 microCi/100 g body weight. From the decay in the specific activity with time after a single injection, the gap junction protein has an apparent half-life of about 19 h. Because of problems of reutilization of radiolabeled amino acid with 35S-methionine as precursor, this apparent half-life probably overestimates the true half-life and indicates a surprisingly rapid turnover of the gap junction protein. This short half-life suggests that, in rat liver, the gap junctions may be very responsive to alterations in physiological demands. PMID- 6273594 TI - Identifying human cells capable of metabolizing various classes of carcinogens. AB - Human cells that appear capable of metabolizing various classes of carcinogens have been identified using one of two methods: metabolism of tritiated benzo(a)pyrene to aqueous-acetone soluble forms or inhibition of cellular DNA synthesis. Each of the assay systems was optimized and the results on 15 human epithelial cell lines were compared. One or more cell lines were found to activate each of four classes of carcinogens examined: polycyclic hydrocarbons, aromatic amines, heterocyclic hydrocarbons, and nitrosamines. Cells that appeared capable of metabolizing polycyclic hydrocarbons or aromatic amines by these methods were also found to produce metabolites which were cytotoxic to cocultivated human xeroderma pigmentosum fibroblasts after a 48-hr exposure to the carcinogen. PMID- 6273595 TI - Double-strand breaks in DNA caused by repair of damage due to ultraviolet light. AB - DNA DSBs are formed in normal human IMR-90 cells during repair incubation after 100 and 300 J.m-2 of UVL. By contrast, no DSBs are formed after UVL in human XPA cells that are unable to excise pyrimidine dimers. The DSBs are not due to immediate cell death since all the cells excluded trypan blue at the time of assay and because XPA cells, which are much more UVL-sensitive than IMR-90, did not form DSBs after UVL. We suggest that these repair-induced DSBs should be potent lesions that might lead to cytotoxicity, chromosome aberrations, deletion mutations, and perhaps cellular transformation. transformation. PMID- 6273596 TI - The structure of fibronectin and its role in cellular adhesion. AB - Fibronectin is a large, adhesive glycoprotein which is found in a number of locations, most notably on cell surfaces, in extracellular matrixes, and in blood. Fibronectin had been detected in all vertebrates tested and in many invertebrates. Its presence in sponges is significant because this suggests that fibronectin may have appeared very early in evolution, possibly with the most primitive multicellular organisms. Cellular and plasma fibronectins have many striking similarities. However, the locations of the polypeptide chain differences between these two proteins indicate that plasma fibronectin cannot be derived from cellular fibronectin by means of simple post-translational proteolysis. Instead, these different types of fibronectin may be products of different genes or of differentially spliced messenger RNA molecules. Amniotic fluid fibronectin is possibly a third form of the protein. Cellular and plasma fibronectins are composed of at least six protease-resistant domains which contain specific binding sites for actin, gelatin, heparin, Staphylococcus aureus, transglutaminase, fibrin, DNA, and a cell surface receptor. The relative locations of these domains have been mapped in the primary structure of fibronectin. The cell surface receptor for fibronectin has not been positively identified, but may be a glycoprotein, a glycolipid, or a complex of the two. Although cell-substratum adhesion is mediated by fibronectin, the locations of the areas of closest approach of the cell to the substratum (the adhesion plaques) and fibronectin are not coincident under conditions of active cell growth. Under conditions of cell growth arrest in low serum concentrations, some fibronectin may become localized at the adhesion plaques. Models describing the domain structure of fibronectin and the molecular organization of the adhesion plaque area are presented. PMID- 6273597 TI - Granular cell tumor in black patients. AB - A 12-year review of granular cell tumors at Howard University Hospital is reported. Sixty-one cases were studied (56 patients). A female preponderance was clearly established and multicentricity of lesions constituted a higher percentage than previously reported in other series. Malignant transformation was not found. Electron microscopic studies tended to support the Schwann cell origin theory for the tumor but no conclusion could be drawn as to whether or not the lesion is neoplastic, degenerative, or metabolic in nature. PMID- 6273598 TI - Perinatal Herpes virus infection: report of a case indicating the paternal role. AB - A case of perinatal Herpes simplex hominis infection with fatal neonatal outcome is reported. The significance of the paternal role is emphasized. It is important to educate sexual partners and stress the serious nature of herpetic infections, especially during the perinatal period. In the presence of active infection, sexual contact should either be totally avoided or a condom should be used. PMID- 6273599 TI - Cytomegalovirus infection of the prostate in the dasyurid marsupials, Phascogale tapoatafa and Antechinus stuartii. AB - Infection by a herpesvirus producing cytomegalic disease in the prostate was demonstrated in the dasyurid marsupials, Phascogale tapoatafa and Antechinus stuartii. The prevalence of lesions among the latter was highest in mature animals during breeding, when the animals are known to be under stress, and in animals treated daily with high levels of exogenous corticosteroid. Occasional cytomegaly was observed in the kidneys of some A. stuartii and may represent a site of latent infection. Virus particles of herpes type were demonstrated by electron microscopy in the nucleus and cytoplasm of infected prostatic cells. It is suggested that the infection may be venereal. PMID- 6273601 TI - Parma wallaby herpesvirus infection. AB - Three Parma wallabies (Macropus parma) were inoculated with a herpesvirus recovered from a captive Parma wallaby with fatal naturally-occurring disease. Two intravenously inoculated animals died after 5 days and one animal infected via the conjunctiva and nasal mucosa was killed when moribund at 7 days. An additional two wallabies held in contact with the others became infected; they were killed at 11 days, when one was severely affected and one was mildly affected. All had small vesicles and ulcers of the skin of the upper and lower lips, eyelids, anogenital area and adjacent genital mucosa. Small vesicles and ulcers and large ulcers, with adherent necrotic epithelium and inflammatory debris, were present on the mucosa of the upper lips and adjacent gums and the conjunctiva. Numerous large basophilic or eosinophilic intranuclear inclusion bodies were observed in the epithelial cells of these vesicles and ulcers and of adjacent hair follicles and sebaceous glands. There was a mild to moderately severe rhinitis. Keratitis was present in two wallabies. Liver lesions were present in two animals but were unlike those seen in herpesviral hepatitis in other species. PMID- 6273600 TI - Serologic survey for selected microbial agents in mammals from Alberta, 1976. AB - Blood samples were taken from humans and several species of free-ranging wild mammals from five different geographic areas of Alberta, Canada. Sera were tested for antibody to eastern equine encephalitis (EEE) virus, western equine encephalitis (WEE) virus, St. Louis encephalitis (SLE) virus, Powassan (POW) virus, the snowshoe hare (SSH) strain of the California group (CAL) of viruses, Northway (NOR) virus, Klamath (KLA) virus, infectious bovine rhinotracheitis (IBR) virus, and two bacteria, Brucella abortus and Francisella tularensis. CAL antibody was found in 63% of 11 snowshoe hares (Lepus americanus), 33% of 167 black bears (Ursus americanus), and 19% of 55 humans (Homo sapiens). NOR antibody was found in 0.4% of 258 hares, 11% of 9 bighorn sheep (Ovis canadensis), 20% of 44 moose (Alces alces), 4% of 56 bears, 14% of 22 woodland caribou (Rangifer tarandus), and 2% of 50 humans. IBR antibody was detected in 14% of 14 moose. B. abortus antibody was found in 1% of 283 bears. F. tularensis antibody was detected in 2% of 52 humans. These findings represent extension of: (1) the natural host range of IBR, CAL, and NOR; (2) the geographical distribution of NOR infection in North America; and (3) the geographical distribution of CAL infection within Alberta. PMID- 6273602 TI - From the NIH: Breast milk and the risk of cytomegalovirus infection. PMID- 6273603 TI - From the NIH: Improved method for diagnosing and monitoring women with gestational trophoblastic neoplasms. PMID- 6273604 TI - Cushing's disease in a patient with an ectopic pituitary adenoma. PMID- 6273605 TI - Cutaneous form of bovine papular stomatitis in man. AB - A cutaneous form of bovine papular stomatitis (BPS) infection was diagnosed in eight persons at the School of Veterinary Medicine at Auburn University, Auburn, Ala. The initial outbreak occurred in five persons who were involved in the care of a bull that required manual placement of an oral feeding tube. Confirmation of diagnosis was based on clinical findings, cytopathological effects in tissue culture, and isolation of typical paravaccinia virus particles in tissue culture. Transmission studies were performed successfully in three normal calves using tissue culture prepared from human biopsy material. In man, the cutaneous form of BPS infection shows gross lesions similar to the cutaneous form of contagious ecthyma ("orf") or pseudocowpox ("milkers' nodules") infection. Because BPS in cattle occurs most often without evidence of readily observable lesions, unlike contagious ecthyma in sheep or pseudocowpox in cattle, the transmission of BPS to man in the cutaneous form could occur without apparent source. The mild clinical manifestations make the condition relatively minor; however, the occasional case may have more severe lesions. PMID- 6273606 TI - Green coffee beans may solve a blood bank problem. PMID- 6273607 TI - Oncogenes. A unitary cause of cancer? PMID- 6273608 TI - [Bioregulation of immunologic function and cyclic nucleotides]. PMID- 6273609 TI - [Immunotherapy and immunochemotherapy of neoplasms-- present status and prospects]. PMID- 6273610 TI - [Blood volume determination of ischemic area with radionuclides]. PMID- 6273611 TI - [Pathology of reticuloendothelial tumors--a consideration from a new point of view]. PMID- 6273612 TI - [2 cases of chemodectoma--catecholamine secreting and non-secreting tumors]. PMID- 6273613 TI - [Pathology of germ cell tumors, with special reference to comparison of various classifications]. PMID- 6273614 TI - [Determination of protein unbound bilirubin by an enzymatic method (author's transl)]. PMID- 6273616 TI - [Experience with scintiangiography of aneurysma dissecans as risk-free examination method without burdening the patient (author's transl)]. PMID- 6273615 TI - [Clinical applications fo reconstructed coronal and sagittal images of body CT (author's transl)]. PMID- 6273617 TI - Epstein-Barr virus-induced lymphoblastoid cell lines from seropositive adult donors and their HLA-antigens. AB - Epstein-Barr virus (EBV)-induced lymphoblastoid cell lines (LCL) were established from seropositive adult Japanese donors. HLA-A, -B, -C and -DR antigens expressed on these EBV-LCL cells were identical to those of peripheral blood lymphocytes. There were no extra reactions upon DR typing; the extra reactivity in A, B and C typing was due to contaminating DR antibodies in the typing sera. In one-way mixed lymphocyte reactions with a stimulator : responder ratio between 1 : 10 and 1 : 20, autologous EBV-LCL exhibited low stimulation which could be distinguished from that of unrelated combinations. EBV-LCL may be useful for the screening of anti-DR antisera and as targets in cell-mediated lympholysis. Furthermore, EBV LCL established from homozygous typing cells may be useful in HLA-D typing. PMID- 6273618 TI - [Scintigraphic detection of small intestinal bleeding - a case of leiomyoma of the ileum (author's transl)]. PMID- 6273619 TI - [Normal pyrimidine 5'-nucleotidase activity level of Japanese subjects and a significance as a marker for low concentration of lead in blood (author's transl)]. PMID- 6273620 TI - The role of NK cells in tumor growth assessed by induction of their suppressor cells in Japanese quails. AB - In Japanese quails treated with chicken amniotic fluid (ChAmF) which had been previously shown to induce suppressor cells to natural killer (NK) cells, tumors appeared with shortened incubation periods after inoculation with Schmidt-Ruppin strain of Rous sarcoma virus (SR-RSV) compared with untreated quails. The tumors in ChAmF-treated quails subsequently grew in a similar pattern to those in untreated quails, whereas by challenging with a lower dose of the virus, enhanced tumor growth was observed as well as earlier onset of tumors in ChAmF-treated quails than in untreated ones. This enhancing effect on tumor growth due to suppression of NK-cell activity was transferred to normal quails with spleen cells obtained from ChAmF-treated quails, since RSV-induced tumors appeared earlier in the recipients of ChAmF-treated spleen cells than in those of untreated spleen cells. These findings show that suppression of NK-cell activity by ChAmF administration rendered quails higher susceptibility to tumor induction by SR-RSV challenge. In other words, NK-cell activity was strongly suggested to contribute to the early protection against tumor growth in the system of Rous sarcoma in Japanese quails. PMID- 6273621 TI - A large-scale epidemic of hand, foot and mouth disease associated with enterovirus 71 infection in Japan in 1978. PMID- 6273622 TI - Smooth muscle relaxing drugs and guinea pig ileum. AB - The effects of various smooth muscle relaxing drugs on contractile responses to acetylcholine (ACh), Ba2+ and Ca2+, and on the tissue cyclic AMP levels were examined in the guinea pig ileum. Papaverine and theophylline caused a decrease both in the maximum height and the slope of dose-response curves induced by the three stimulants, and an increase in the cyclic AMP levels. Diltiazem and D-600 produced a decrease in the maximum and the slope of ACh and Ba2+ dose-response curves, shifted the Ca2+ dose-response curves to higher concentrations, in a parallel manner, but failed to change the cyclic AMP levels. Etomidoline and benactyzine shifted the curves for the three stimulants in parallel to the right, but at higher concentrations depressed the maximum of ACh and Ba2+ responses with a further parallel shift. These drugs exerted little influence on the basal level of tissue cyclic AMP, but etomidoline significantly depressed the Ba2+ -induced increase in cyclic AMP level. The smooth muscle relaxing drugs used could be classified in three types, thereby suggesting that there are at least three different mechanisms involved in smooth muscle relaxing action. PMID- 6273623 TI - Inhibition of adenylate cyclase by GTP and its modulation by opiate receptor in rat caudate nucleus. AB - Morphine inhibited the adenylate cyclase activity of the crude synaptosomal fraction of the rat caudate nucleus in the presence of BTP, GDP, Gpp(NH)p or ITP. The purine nucleotides themselves had an inhibitory action on the enzyme. Beta endorphin and Met-enkephalin also inhibited the enzyme in the presence of GTP. The GTP-dependent in inhibitory action of morphine was blocked by naloxone. Various opiates and opioid peptides inhibited the enzyme by up to approximately 20 per cent in the presence of GTP. The relative potency was in higher order of levorphanol greater than beta-endorphin greater than Met-enkephalin greater than morphine greater than pentazocine. Levorphanol was about 50,000 times as potent as its biologically inactive enantiomer, dextrorphan. Morphine enhanced the inhibitory actions of GTP and GTPase-resistant Gpp(NH)p on the adenylate cyclase activity. These results suggest that GTP plays an important role in the regulation of adenylate cyclase activity in the rat caudate nucleus and that the occupation of opiate receptor by agonists inhibits the enzyme through an actual increase in the inhibitory action of GTP, rather than a suppression of the enzymatic degradation of GTP. PMID- 6273624 TI - Effect of hypothyroid status on adenosine 3',5'-monophosphate-dependent protein kinase of rat kidney. AB - Changes in the properties of cyclic AMP-dependent protein kinase were studied in the kidney of hypothyroid rats. The activity of the crude extract was greater in hypothyroid rats than in normal ones. The ratio of type I isozyme to type II isozyme in hypothyroid rats was about twice that in the normal rats with enzymes from both the cortex and medulla. A higher affinity for the substrates was shown in the enzymes of the hypothyroid rat kidney. The effects of actinomycin D and of heat-stable protein kinase inhibitor on the enzyme activity were the same in both normal and hypothyroid rats. The changes in enzyme properties and isozyme distribution may well play an important role in the hypothyroid rat kidney. PMID- 6273625 TI - Effects of electroconvulsive shock on mouse-killing behavior (muricide) in olfactory bulbectomized rats. AB - We investigated the influence of electroconvulsive shock (ECS), regarded to possess an antidepressant effect clinically, on muricide in olfactory bulbectomized rats (OB rats). Muricide in these rats was markedly inhibited by ECS treatment. Five and 10 min after the termination of ECS-induced convulsions, muricide was inhibited by 100%. Even after intervals of 20 and 60 min, inhibition rates of 80% and 30% were obtained, respectively. ECS-induced muricide inhibition was remarkably antagonized by pretreatment with the alpha-blocker phenoxybenzamine but not by pretreatment with the beta-blocker sotalol. ECS induced suppression of muricide was potentiated by repeated ECS treatment once daily for 10 days. After several applications of ECS treatment, muricide was inhibited in muricide tests done 24 hours after ECS treatment; this state persisted for up to 10 days thereafter. The results of this experiment demonstrated that ECS treatment specifically inhibited muricide in OB rats and further suggested that the cerebral noradrenergic alpha-receptor system plays an important role in this ECS-induced inhibition of muricide. Similar findings in the case of antidepressant administration, inhibition of muricide was potentiated by chronic ECS treatment. Specific inhibition of muricide in OB rats by antidepressants indicated that this phenomenon may serve as an animal model for the evaluation of antidepressant activity. Our results reiterate the usefulness of muricide in OB rats as an excellent experimental model for the assessment of antidepressant activity. PMID- 6273626 TI - Anti-hypertensive effects of propranolol unaltered by 6-hydroxydopamine given intraventricularly to SHR. PMID- 6273627 TI - Pharmacological studies on BB-1502, a new bronchodilator. AB - The pharmacological activities of BB-1502 (9-cyclohexyl-2-n-propoxy-9H-adenine), a potent bronchodilator and inhibitor of cyclic AMP phosphodiesterase, were compared with those of aminophylline in a number of biological systems. IOn vitro, BB-1502 relaxed guinea pig tracheal tissue at a concentration ca. 600 times lower than that required for aminophylline. The ability of BB-1502 to inhibit cyclic AMP phosphodiesterase of guinea pig lung origin was 15 times greater than that of aminophylline. The direct bronchodilator activity of BB-1502 determined in guinea pigs by the intraduodenal route was 5 times greater and the duration of the action longer than that of aminophylline. In dogs, intraduodenal BB-1502 inhibited bronchoconstriction induced by pilocarpine, histamine, or acetylcholine and the activity of the new compound was 4 to 7 times more potent than that of aminophylline. In experimental asthma studies, orally administered BB-1502 protected guinea pigs from allergy-induced asthma provoked by an antigen challenge with aerosolized egg albumin and here the potency was 4 times greater that that of aminophylline. Ascaris antigen-induced asthma in dogs was completely inhibited by the oral administration of BB-1502 in a dose of 1 mg/kg with significant protection seen at 0.3 and 0.1 mg/kg. No complete protection was obtained with aminophylline in the dog asthma model. Cardiovascular effects observed with guinea pigs and dogs following the oral administration of BB-1502 were considerably less than those seen with aminophylline. CNS side effects were nil in rats when BB-1502 was given at 3 times the dose that produced significant stimulation by aminophylline. Acute lethal toxicity determined in mice by the oral route was similar for BB-1502 and aminophylline. PMID- 6273628 TI - Role of alpha-adrenergic receptors in denervation supersensitivity of rat vas deferens. AB - Contractile responses of rat vas deferens were studied with particular attention directed to the role of receptors and neuronal control. Marked contraction of the vas deferens was observed with alpha-adrenergic agonists, depending on their concentrations. This tissue had a low sensitivity to ACh. Four days after denervation, this tissue showed a supersensitivity to alpha-adrenergic agonists and a high K+ concentration, but not to ACh. The increase in sensitivity to alpha agonists resulted in an enhancement of the maximal response and a shift of the concentration response curve to lower concentrations of these reagents. Alterations were seen in the alpha-adrenergic receptors in the rat vas deferens, assayed by measuring the binding of [3H]WB4101. The maximal binding sites decreased significantly to 86 from 142 fmoles per mg protein. The affinity of the receptors for alpha-agonist, determined by measuring the ability of agonists to displace bound [3H] WB4101, increased significantly, while the affinity to alpha antagonists remained unchanged. Studies on [3H]QNB binding indicated no significant change in muscarinic ACh receptors after denervation. Thus, supersensitivity of the alpha-adrenergic mechanism mediated by a specific change in affinity of alpha-receptors occurs after denervation of rat vas deferens. These changes in sensitivity and in receptors are discussed in relation to the characteristics and roles of alpha-receptors in the rat vas deferens. PMID- 6273629 TI - Effects of YS-980, an orally active converting enzyme inhibitor, on blood pressure in normotensive and hypertensive rats. AB - The hypotensive effect of YS-980, (4R)-3-[(2S)-3-mercapto-2-methyl propanoyl]-4 thiazolidinecarboxylic acid, was investigated in normotensive and different models of hypertensive rats. Experiments were carried out in both anesthetized rats (1 mg/kg i.v.), and conscious unrestrained rats (10 mg/kg p.o.) YS-980 markedly lowered blood pressure in acute renal hypertensive and two-kidney, one clip hypertensive rats and moderately lowered the pressure in SHR. Contrary to these hypotensive effects, this agent did not reduce the blood pressure in DOCA hypertensive rats. In normotensive rats, YS-980 had a slight hypotensive action in anesthetized rats but not in the conscious animals. The hypotensive effect of YS-980 is attributed mainly to suppression of the renin angiotensin system. PMID- 6273630 TI - A possible mechanism for relaxation of rat uterine smooth muscle by nicardipine hydrochloride (YC-93), a new potent vasodilator. AB - A new potent vasodilator, nicardipine hydrochloride inhibited oxytocin-induced contraction of rat uterus dose-dependently with an increase in the intracellular cyclic AMP level at the onset of relaxation. Dibutyryl cyclic AMP and papaverine, an inhibitor of cyclic AMP phosphodiesterase (PDEase), also inhibited the contraction. Nicardipine inhibited competitively PDEase in homogenates of rat uterus which exhibited apparently two Km values for cyclic AMP (3.6 micro M and 67.3 micro M) with the Ki of 5.3 micro M and 13.2 micro M, respectively, but had no effect on adenylate cyclase. Nicardipine enhanced calcium uptake by rat uterine microsomes, at concentrations which inhibited oxytocin-induced contraction in the same manner as cyclic AMP. The maximal stimulation by nicardipine of the microsomal calcium uptake was identical substantially to that by cyclic AMP, and both were not additive. Cyclic AMP was also accumulated during the uptake reaction in the presence of nicardipine. On the contrary, neither myosin ATPase nor microsomal Ca2+-dependent ATPase was inhibited directly by nicardipine. These results suggest that the inhibition of oxytocin-induced contraction of rat uterus by nicardipine may be due to an enhancement of microsomal calcium uptake, mediated by cyclic AMP accumulated through the inhibition of PDEase. PMID- 6273631 TI - Further studies on the synthesis of A-form monoamine oxidase. AB - The relation between precursors and restoration of A-form MAO activity in rat liver after administration of clorgyline to rats was investigated by measuring the rates of recovery of A-form MAO activity after treatment with the inhibitor. The half-lives of mitochondrial and microsomal A-form MAO were estimated as 3.5 and 2.0 days, respectively. MAO activity and the amount of MAO molecules were completely restored within 14 days. However the values attained did not exceed the control values in a period of 14 days. Clorgyline plus cycloheximide or chloramphenicol did not prevent the recovery of MAO activity in the microsomes, but did not delay the appearance of enzyme activity in the mitochondria. A and B form-like MAO were also observed in the microsomal and supernatant fractions, with clorgyline as inhibitor. These results suggest that the microsomal enzyme is a precursor of the mitochondrial enzyme, that the levels of A-form and B-form MAO are regulated genetically, and that the two forms of MAO may be synthesized separately. PMID- 6273632 TI - Mitotic index in gastric carcinoma of the scirrhous type. AB - For the purpose of studying the clinical and pathological specificities of the scirrhous type in gastric carcinoma, 96 patients with gastric carcinoma were studied according to age, sex, stage, macroscopic types and mitotic index. Scirrhous carcinoma was diagnosed in 45 patients. There were higher incidences of scirrhous carcinoma in the younger generation and among women. In the scirrhous type, the percentage of stages III and IV was higher than that of stages I and II (p less than 0.005). In Borrmann's categories III and IV, patients with the scirrhous type were more numerous than those with the other types. Mitotic indices were 18.4 +/- 8.1 in scirrhous type, 52.8 +/- 13.5 in medullary type, 54.1 +/- 16.9 in intermediate type and 53.3 +/- 11.5 in non-cancerous gastric mucosa. The low rate of mitotic index in those with scirrhous carcinoma was statistically significant compared to those of other types and that of the non cancerous gastric mucosa (P less than 0.001). Mitotic index is apparently related to the extent and the degree of degeneration of interstitial connective tissue. The speed of the growth of the tumor mass in gastric carcinoma is not so closely related to the mitotic index, and relates to a greater extent the number of dissociated cells entering the digestive canal. PMID- 6273633 TI - Local immunoglobulin A in normal and neoplastic breast tissues. AB - Local IgA levels in 36 normal, 92 nonmalignant atypical, and 47 malignant human breast tissue specimens were analyzed by three methods: direct immunofluorescence, modified radioimmunoassay for IgA in cryostat sections, and the Immuno-Fluor (Bio-Rad Laboratories, Richmond, Calif.) assay for the cell surface IgA in tissue eluates. The proportion of epithelium in each sample was estimated microscopically. For malignant specimens the proportion of epithelial cells was significantly higher than that for nonmalignant specimens. The immunofluorescence test showed no qualitative difference in epithelial IgA content between normal and nonmalignant atypical tissues, but the frequency of positive specimens was markedly lower in malignant tissues. The modified radioimmunoassay and the Immuno-Fluor assay indicated little or no difference in IgA concentration per tissue weight among normal, nonmalignant atypical, and malignant breast tissues. However, when the data were corrected for proportion of glandular epithelium, nonmalignant atypical tissues had less IgA per cell than normal tissues, and malignant mammary carcinomas had significantly less IgA per cell than both normal and nonmalignant atypical tissues. PMID- 6273634 TI - Spontaneous lymphomas in SJL/J-(v+) mice: ecotropic and dualtropic virus expression in normal and lymphomatous tissues. AB - Approximately 60% of inbred SJL/J-(v+) adult mice having high levels of ecotropic endogenous XC+ virus showed virus activation within the first month of life, while the others produced virus at comparable levels later on, in an attempt to correlate the time of virus activation with the incidence and latency of lymphomas, the tails of 57 1- and 2-month-old mice were tested for virus presence, and the mice were then observed for lymphoma appearance. While all 2 month-old mice expressed ecotropic virus, only 63% of the 1-month-old mice were virus-positive. However no relationship existed between early virus production (within 1 month) or late virus production and lymphoma latency, total lymphoma incidence, and histopathology. In contrast with high titers of XC+ virus in tail tissues of diseased mice, a markedly low virus content was found in lymphomatous organs. This difference was not due to selective growth of poor virus-producer cells or to inhibitory factors possibly released by the inflammatory cell component. Viral protein content and XC+ virus titer were not closely correlated in the neoplastic organs. Search for XC- viruses revealed that only 1 of 6 aged normal and 16 of 19 lymphomatous mice produced viruses that grew on mink lung cells. By use of a standard limiting dilution cloning procedure, four isolates were obtained that showed tropism for both murine and heterologous cells. Three of these isolates induced cytopathic changes similar to those induced by MCF viruses on mink lung cells but not on mouse cells. Interference and neutralization assays performed to better characterize the virus envelope properties further indicated that SJL/J isolates had features typical of MCF dualtropic viruses. PMID- 6273635 TI - Inhibition of adenovirus oncogenicity in hamsters by adeno-associated virus DNA. AB - Adeno-associated virus (AAV) DNA partially inhibited in Syrian golden hamsters the formation of tumors by the human adenovirus (Ad) type 12. Furthermore, defective interfering AAV particles or their DNA also reduced tumorigenesis. Defective AAV particles contain aberrant genomes with extensive deletions of the internal AAV DNA sequences. Variant AAV DNA, containing 30% of the viral genome, decreased the incidence of Ad-induced tumors from 44 to 18%. Defective AAV particles, which have a buoyant density of 1.32 g/cm3 in CsCl and which are highly enriched for the DNA of the terminal regions (map positions, 0-5 and 95 100), completely suppressed Ad oncogenicity. This observation suggested that the AAV DNA sequences close to the terminal region of the genome mediated the inhibition of the Ad oncogenicity. PMID- 6273636 TI - Enhancement of experimental pancreatic cancer in Syrian golden hamsters by dietary fat. AB - The effects of dietary fat on the induction and development of pancreatic ductular adenocarcinoma were studied in randombred Syrian golden hamsters. Diets containing low-fat (LF) or high-fat (HF) levels of corn oil [4.5 or 18.0 g/385 kilocalorie (kcal)], contributing 10 or 41% of the calories, respectively, were fed either before or after a single injection of N-nitrosobis(2-oxopropyl)amine (BOP) (10 mg/kg body wt). Control hamsters were fed corn oil at a medium-fat (MD) level (9 g/385 kcal) for life. The incidence of ductular adenocarcinomas increased in both males and females (LF diet, 16%; HF diet, 34%) when the HF diet was fed after BOP treatment. The average number of carcinomas per carcinoma bearing animal also increased (LF diet, 1.3; HF diet, 3.0), but the carcinoma incidence was not influenced by these diets being fed before carcinogen treatment. The incidence of ductular adenomas was high with all treatments and was not influenced by diet. However, the number of adenomas was increased in animals fed HF diets. In addition, the incidence of acinar cell nodules was elevated in animals fed the MF and HF diets after BOP administration. These results showed that dietary fat modified the development of experimental ductular adenocarcinoma of the pancreas. PMID- 6273637 TI - Age- and sex-related differences in antibody formation and blastogenic responsiveness of splenocytes from RIII mice developing virus-induced mammary adenocarcinoma. AB - Inbred RIII mice, known to be infected neonatally with murine mammary tumor virus so that females develop mammary adenocarcinoma by 12-15 months of age, were examined with regard to antisheep red blood cell antibody responses at the cellular level. Female mice, 3-11 months old, compared to male mice of the same ages had consistent and significant depression of the antibody response of their splenocytes. Furthermore, female mice with adenocarcinoma showed an even greater depression of the antibody response. Spleen sizes were consistently increased in females as compared to those of male mice throughout the first year of life. The blastogenic responsiveness of the splenocytes to the B-cell mitogen Escherichia coli lipopolysaccharide and the T-cell mitogen phytohemagglutinin-P was not significantly different between male and female mice during the same periods, although the responses of the older tumor-bearing female mice to phytohemagglutinin-P were lower than those of non-tumor-bearing female mice. A complex relationship between age, sex, and immune responsiveness was evident in these mammary tumor virus-infected mice, which made it difficult to attribute a specific immune event to emergence of the mammary adenocarcinomas in the female as compared to male mice. PMID- 6273638 TI - Cytotoxic effects and metabolism of benzo[a]pyrene and 7,12 dimethylbenz[a]anthracene in duodenal and ileal epithelial cell cultures. AB - Two epithelial cell lines have been established from the duodenum (IEC-17) and the ileum (IEC-18) of outbred germfree Crl:CD(SD)GN rats. They have a very similar morphology and ultrastructure, a normal rat diploid karyotype, comparable growth rates, and a similar set of surface antigens as detected with monoclonal antibodies specific for intestinal epithelial cell surface proteins in vivo. Both cell lines do not grow in soft agar and when injected into syngeneic animals do not form tumors. The toxic effects of benzo[a]pyrene (BP) and 7,12 dimethylbenz[a]anthracene (DMBA) were studied in these two cell lines. The IEC-18 cells were found to be much more sensitive to both chemicals than the IEC-17 cells. In particular, at concentrations greater than 1 microgram/ml, DMBA completely and irreversibly inhibited cell proliferation in the IEC-18 cells, whereas it produced a less marked and reversible inhibition in the IEC-17 cells. The metabolism of BP and DMBA to water-soluble products was studied; BP was apparently metabolized at similar rates in the two cell lines, whereas a striking difference was observed with DMBA, which was metabolized at a rate 10-15 times greater in the IEC-18 cells than that in the IEC-17 cells. These results suggest that cultured cell lines derived from different portions of the intestinal tract have similar properties in vitro, but they may have very different responses to chemical carcinogens. PMID- 6273639 TI - Immunogenicity of guinea pig cells transformed in culture by chemical carcinogens. AB - The immunogenicity of inbred strain 2/N guinea pig fibroblasts transformed to the malignant state in vitro by chemical carcinogens was evaluated with the use of a variety of in vivo and in vitro methods including delayed-type hypersensitivity skin and tumor transplantation tests and analysis of antibody production by immunofluorescence, complement fixation, and staphylococcal protein A binding tests. Neoplastic transformation was induced by direct treatment of cells in culture with benzo[a]pyrene, 3-methylcholanthrene, or N-methyl-N'-nitro-N nitrosoguanidine (MNNG) or by the host-mediated method by which fetuses were exposed to diethylnitrosamine or MNNG in vivo prior to cell culture. Rabbits and syngeneic guinea pigs were inoculated with unirradiated and X-irradiated clonally derived cells. Delayed hypersensitivity skin reactions to immunizing or other cells were equivalent in immunized or control guinea pigs, and no protection to tumor outgrowth from a challenge inoculum of immunizing cells was observed. Antibody activity induced in the sera of immunized guinea pigs was cross-reactive and removed by absorption with nontumorigenic cells. Rabbit antisera after absorption with fetal guinea pig cells were nonreactive with the specific immunizing or other culture cells. Chemical carcinogen-induced neoplastic transformation of guinea pig cells can, therefore, occur without formation of detectable, individually distinct cell surface tumor-specific neoantigens. PMID- 6273640 TI - Clinical conferences at the Johns Hopkins Hospital. Klatskin tumor: radiologic features of biliary tract disease. PMID- 6273641 TI - [A case of small-cell carcinoma misdiagnosed as lung tuberculosis in consequence of shadow regression during initial administration of rifampicin (author's transl)]. PMID- 6273642 TI - [Significance of calcium metabolic disorders in myocardial pathology]. PMID- 6273643 TI - Mechanisms of action of various hormones and vasoactive substances on glomerular ultrafiltration in the rat. AB - Angiotensin II (AII) and arginine vasopressin are capable of triggering glomerular mesangial cell contraction in vitro. A similar mechanism acting in vivo to reduce glomerular capillary surface area could account for the decline in the ultrafiltration coefficient (Kf)( that occurs in single glomeruli in response to infusion of these substances. Less clear is the mechanism whereby similar declines in Kf are induced with infusions of dibutyryl cyclic AMP (DBcAMP), parathyroid hormone (PTH), and prostaglandins, because PTH and PGE2, at least, are incapable of eliciting mesangial cell contraction in vitro. To further explore the factors that regulate Kf in vivo, we performed micropuncture experiments in 47 euvolemic Munich-Wistar rats. Infusions of DBcAMP, PTH, prostaglandins I2 and E2 led to lower mean values for plasma flow rate (QA) and Kf in superficial glomeruli than were found in animals given vehicle alone (control group), whereas average values for glomerular transcapillary hydraulic pressure difference (delta P) and total renal arteriolar resistance (RTA) tended to be higher. These increases in delta P and RTA, and decreases in QA and Kf, with DBcAMP, PTH, PGI2, and PGE2 are typical of changes induced by AII. Indeed, when saralasin, a competitive AII antagonist, was infused together with these various vasoactive substances, the effects on delta P, QA, RTA, and Kf were largely abolished. Therefore, the actions of DBcAMP, PTH, PGI2, and PGE2 on the glomerular microcirculation appear to depend on an intermediate action of AII. By contrast, although pitressin (ADH) infusion also led to a significant decline in Kf, saralasin administration did not reverse this change, suggesting that the action of ADH on the glomerular microcirculation is independent of a pathway involving AII. Based on these studies, it seems reasonable to propose that AII and ADH are both potentially important regulators of mesangial cell contraction, and thereby, glomerular capillary filtering surface area and Kf. PMID- 6273644 TI - [The C-peptide suppression test in normal persons and insulinoma patients: an attempt to evaluate its use in functional diagnosis (author's transl)]. PMID- 6273646 TI - [Emergency states in diseases of the peripheral nervous system]. PMID- 6273645 TI - Natriuretic hormone - a circulating inhibitor of sodium- and potassium-activated adenosine triphosphatase. Its potential role in body fluid and blood pressure regulation. AB - Expansion of the extracellular fluid volume (ECFV) promotes the release of a small molecular weight (less than or equal to 1,000 Daltons) humoral natriuretic factor. The resulting natriuresis is accompanied by inhibition of renal cortical tissue and red blood cell Na-K-ATPase activity. This transport inhibitor, presumably an acidic peptide derived from a larger precursor molecule, was so far recovered from the serum and urine of rat, dog, and man, and from renal cortical tissue homogenate. Using Sephadex G-25 gel chromatography the inhibitor is eluted in the post-salt fraction IV. Its natriuretic action is demonstrated by bioassay methods, it depresses sodium transport of isolated amphibian membranes and inhibits Na-K-ATPase enzyme activity in vitro. The inhibitor isolated from human urine binds to specific antibodies against digoxin. This natriuretic factor is absent in patients with arterial hypotension and in edematous patients with secondary aldosteronism. In contrast, high inhibitory activity is found in patients with primary aldosteronism, a condition which represents the most classical type of low renin volume-dependent hypertension. Since enzyme inhibition by this humoral endogenous agent probably extends to various Na-K ATPase-dependent transport systems including vascular smooth muscle fibers, depression of their Na-K pump will raise intracellular concentrations of sodium and calcium and thereby induce vasoconstriction. It is therefore tempting to speculate that the natriuretic hormone plays an important role in the pathogenesis of volume-dependent arterial hypertension. PMID- 6273647 TI - [Succinate dehydrogenase and cytochrome oxidase activity in rat tissues during prolonged hypokinesia]. AB - The activity of cytochrome oxidase in the liver, skeletal muscles, kidneys, brain and lungs of 56 white rats decreased on days 15 and 60 of the hypokinetic study. The activity of succinate dehydrogenase significantly lowered in the brain and slightly diminished in the lungs and skeletal muscles on the same days. The activity of succinate dehydrogenase in the liver increased on the 90th hypokinetic day. PMID- 6273648 TI - Intranasal vaccine trial for canine infectious tracheobronchitis (kennel cough). AB - Two field trials were conducted during periods of endemic (summer) and epizootic (winter) canine infectious tracheobronchitis activity to evaluate the efficacy of three intranasal vaccines in a closed commercial beagle breeding kennel. A trivalent vaccine containing Bordetella bronchiseptica, canine parainfluenza, and canine adenovirus-2 was administered at 3 weeks of age. The vaccine was 71.2% and 81.8% effective in decreasing the incidence of coughing during the winter and summer trials, respectively. The number of deaths was lower in each of the vaccine groups than in the placebo groups. No adverse reactions were observed with any of the intranasal vaccines. PMID- 6273649 TI - Fibroblast alpha-galactosidase A activity for identification of Fabry's disease heterozygotes. AB - The identification of female carriers of Fabry's disease is important for genetic counselling since prenatal diagnosis of affected fetuses is possible. The activities of either total alpha-galactosidase or alpha-galactosidase A in cultured fibroblasts were similar in Fabry carriers and controls and cannot therefore be used for carrier detection. Better discrimination between carriers and controls was found when total alpha-galactosidase activity was expressed as a ratio to beta-galactosidase activity, but overlap still occurred. However, there was complete discrimination between the ratio of alpha-galactosidase A to beta galactosidase in cultured fibroblasts from five carriers of Fabry's disease and either 11 controls, seven hemizygote affected males or two of their female relatives. PMID- 6273650 TI - A new variant of glycogen storage disease type 1: probably due to a defect in the glucose-6-phosphate transport system. AB - A new variant of glycogen storage disease (GSD) Type 1, with clinical symptoms and laboratory findings consistent with those of glucose-6-phosphatase (G6Pase) deficiency, is described. Assay of G6Pase in liver from the patient immediately after biopsy by the method of Nordlie and Arion gave low activity (0.8 mumol/min per g liver) in the absence of detergent, but was normal (10.2 mumol/min per g liver) after addition of detergent. Liver stored for a day at -25 degrees C had normal activity (3.4 mumol/min per g liver) without detergent. In patients with GSD Type la, G6Pase activity was very low both with and without detergent. These findings suggest a defect in glucose-6-phosphate transport in the microsomal membrane of the patient's liver. The integrity of microsomal membrane was destroyed by storage at -25 degrees C, when activity of G6 ase in the patient's liver could be demonstrated. This may be the first example of a disorder involving the transport system of an intracellular membrane. PMID- 6273652 TI - Use of monofilament polyglycolic acid suture for experimental peripheral nerve repair. PMID- 6273651 TI - Neutral 17beta-hydroxysteroid oxidoreductase deficiency in testes causing male pseudohermaphroditism in an infant. AB - A deficiency of neutral 17beta-hydroxysteroid oxidoreductase activity in testes has been diagnosed in a n infant with male pseudohermaphroditism. In vivo stimulation tests of testicular endocrine function with human chorionic gonadotrophin provided an accurate diagnosis in contrast to estimates of enzymic activity in vitro in testes and other tissues. The discrepancy in testes may be due to the absence of gonadotrophin stimulation in the latter studies. The in vitro studies show that there are at least two forms of 17beta-hydroxysteroid oxidoreductase under independent genetic control and that only one form is localized to the testes. The diagnosis before puberty has allowed early treatment by removal of the abnormal testes which should prevent the usual presenting clinical signs of marked masculinizatin and hirsutism at puberty. PMID- 6273654 TI - AMA 1980 interim meeting report. Marijuana in the '80s. PMID- 6273656 TI - Effect of ACTH on the subcellular distribution of steroids in the adrenal gland. PMID- 6273653 TI - Cellular and subcellular aspects of the mechanism of gastric acid secretion. PMID- 6273655 TI - Mineralocorticoid binding in cultured smooth muscle cells and fibroblasts from rat aorta. PMID- 6273657 TI - Histological and epidemiological features of breast cancer in different religious groups in greater Bombay. AB - In this paper an attempt has been made to study the histologic and epidemiologic features of breast cancer in women from various religious groups of Greater Bombay. The crude and age-adjusted incidence rates are seen to be the highest in the Parsi community followed by the Christian, Moslem, and Hindu groups. In all the religious groups, an increase in incidence of breast cancer is first seen in the third decade. This is followed by a sharp increase up to the age of 50-54, leading to a slight drop, and then a consistent but slower rise in the older ages. The high risk of developing this cancer in Parsi women as compared with the Christian, Moslem, and Hindu experience may be due to a higher proportion of Parsi women remaining unmarried, their higher age at marriage, lower age at first pregnancy, broad spacing of pregnancies, and fewer numbers of pregnancies. The distribution according to histologic types of breast cancer in India is characterized by a high frequency of infiltrating duct carcinoma. Medullary, lobular, and squamous cell carcinoma were typical histological varieties of the older age groups, whereas ductal carcinoma was encountered mostly in premenopausal women. PMID- 6273658 TI - Method for measuring the duration of inhibition of angiotensin I-converting enzyme in vivo. AB - The spontaneously hypertensive rat, either pithed or anesthetized with urethane, responds to intravenous injections of angiotensin I (A1) and angiotensin II (AII) with pressor responses. These responses vary with time, but the ratio of the responses to AI and AII is constant throughout 4.5 hr. This criterion was used to determine the degree and the duration of effect of captopril. At low (30 micro g/kg iv) and high (3mg/kg iv) doses, it selectively reduces the pressor response to AI, so reducing the AI/AII ratio. The rate of recovery is initially rapid followed by a prolonged slower phase. The degree (maximum 65%) and the duration of inhibition are directly related to the dose of the inhibitor. PMID- 6273659 TI - Gestational trophoblastic neoplasms. Clinical principles of diagnosis and management. PMID- 6273660 TI - Pathology of trophoblastic disease. PMID- 6273661 TI - The clinical nurse specialist's role in the management of gestational trophoblastic neoplasms. PMID- 6273662 TI - Psychological and social impact of gestational trophoblastic neoplasms. PMID- 6273663 TI - Measurement of human chorionic gonadotropin. PMID- 6273664 TI - Cytomegalovirus infections: current diagnostic methods. PMID- 6273665 TI - Myelin-like inclusions in maturing and senescent muscle cells of rat myocardium. AB - This paper deals with the ultrastructure of two types of cytoplasmic inclusions, "myelin figures" and "lipid complexes", present in muscle cells of ventricular myocardium of the rat heart. The inclusions were numerous in both the first two postnatal weeks and in the senescent stages, being rarely observed in young adult animals. The myelin figures were entirely made up of myelin-like membrane whorls, whereas the lipid complexes exhibited discrete myeloid caps surrounding a central lipid core. The origin and functional significance of such myelin-like structures are discussed on the basis of ultrastructural and biochemical studies on myocardial physiology and age-related evolution. PMID- 6273666 TI - Metabolic activation of aflatoxin B1 by liver tissue from male fischer F344 rats of various ages. AB - The effect of aging on the ability of the liver to activate chemical procarcinogens was studied using 12-, 18-, and 27-month-old male Fischer F344 rats. The cytochrome P-450 content of the S9 and microsomal fractions of the liver decreased approximately 30% between 12 and 18 months of age. The structural conformation of cytochrome P-450 in microsomes from 12-, 18-, and 27-month-old rats was studied using electron-spin resonance spectroscopy. An age-related decrease in the amount of cytochrome P-450 ferric iron in the liver microsomes was observed. The conversion of the chemical procarcinogen aflatoxin B1 to mutagenic compounds by the S9 and microsomal fractions of liver was measured using the Salmonella typhimurium bioassay. A 40-50% decrease in the metabolic activation of aflatoxin B1 was observed between 12 and 18 months of age. However, the activation of aflatoxin B1 did not change after 18 months of age. The age related decrease in the activation of aflatoxin B1 by liver appears to be due to a decrease in the metabolic activity of the mixed-function oxidase system. PMID- 6273667 TI - Slow viral infections of animals: experimental models for human disease. AB - Sigurdsson's three criterions for a slow viral infection are quoted and discussed and the background of his work briefly described. A fourth criterion for a slow viral infection, is suggested, infection of the hosts lymphoid tissues, which is a common feature of slow infections that have been observed. The general properties of the maedi-visna virus, the diseases and the immune response it causes are discussed. Sheep and goats are susceptible to natural maedi-visna infection. In the goat an identical retrovirus causes arthritis. Arthritis has not been found in maedi-visna infected sheep. Two subacute spongiform encephalopathies of animals are shortly reviewed, scrapie in sheep and goats and transmissible mink encephalopathy (TME). General properties of the very unusual scrapie agent are mentioned briefly. The third type of a slow viral infection is mentioned, Aleutian mink disease, an immunopathological disorder of certain minks caused by a selective infection of lymphoid cells. PMID- 6273668 TI - The relation of the soluble thiamine triphosphatase activity of various rat tissues to nonspecific phosphatases. AB - Polyacrylamide gel electrophoresis was used to investigate the relation of the soluble thiamine triphosphatase activity of various rat tissues to other phosphatases. This technique separated the thiamine triphosphatase of rat brain, heart, kidney, liver, lung, muscle and spleen from alkaline phosphatase (EC 3.1.3.1), acid phosphatase (EC 3.1.3.2) and other nonspecific phosphatase activities. In contrast, the hydrolytic activity for thiamine triphosphate in rat intestine moved identically with alkaline phosphatase in gel electrophoresis. Thiamine triphosphatase from rat liver and brain was also separated from alkaline phosphatase and acid phosphatase by gel chromatography on Sephadex G-100. This gave an apparent molecular weight of about 30,000 and a Stokes radius of 2.5 nanometers for brain and liver thiamine triphosphatase. The intestinal thiamine triphosphatase activity of the rat was eluted from the Sephadex G-100 column as two separate peaks (with apparent molecular weights of over 200,000 and 123,000) which exactly corresponded to the peaks of alkaline phosphatase. The isoelectric point (pI) of the brain thiamine triphosphatase was 4.6 (4 degrees C). The partially purified thiamine triphosphatase from brain and liver was highly specific for thiamine triphosphate. The results suggest that, apart from the intestine, the rat tissues studied contain a specific enzyme, thiamine triphosphatase (EC 3.6.1.28). The specific enzyme is responsible for most of the thiamine triphosphatase activity in these tissues. Rat intestine contains a high thiamine triphosphatase activity but all of it appears to be due to alkaline phosphatase. PMID- 6273669 TI - [Inhibitory effect of ethyl ester of 2-(4-(chlorophenoxy) isobutyric acid "clofibrate" on Aujeszky's disease virus synthesis in cell cultures]. PMID- 6273670 TI - Cataracts related to enzymes of galactose metabolism. PMID- 6273671 TI - Tapetoretinal degeneration and mental retardation associated with microspherophakia and mesodermal abnormalities: a new syndrome? PMID- 6273672 TI - [Lung resections in bronchial carcinoma of patients older than 70 years (author's transl)]. AB - In a retrospective study (1969--1979) of 530 resections performed because of bronchial carcinoma, there were 70 patients older than 70 years (13.2%). Two thirds of their resections were carried out on T 1--2 N0M0 and one-third on T 1- 2-N1M0. For 22 central and 48 peripheral carcinomas, 25 pneumonectomies, 40 lobectomies, and 5 bilobectomies were performed. The mortality was 14.2% (lung embolism, pneumonia, apoplexia, and one insufficiency of the bronchial resection stump). Postoperative complications were atelectasis (28.5%), pneumonia (4%), lung embolism (6%), and two pleura empyemas. Of the patients operated on from 1976 to 1978, 40% (T1N0M0) and 30% (T2N0M0) and 30% (T2N0M0) are alive after 2.5 years. PMID- 6273673 TI - Immunopathology of nerve damage in leprosy. PMID- 6273674 TI - Comparison of the muscarinic-cholinergic and lysophosphatidate inhibition of fibroblast adenylate cyclase demonstrating desensitization to the cholinergic stimulus. PMID- 6273675 TI - Amphetamine effects on acetylcholine release and mammalian neuromuscular transmission. PMID- 6273676 TI - Beta-endorphin immunoreactivity in rat plasma: variations in response to different physical stimuli. PMID- 6273677 TI - Properties of [3H] beta-carboline-3-carboxylate ethyl ester binding to the benzodiazepine receptor. PMID- 6273678 TI - Barbiturate and GABA receptors coupled to benzodiazepine receptors in rat hippocampal formation: a radiohistochemical study. PMID- 6273679 TI - The effect of ACTH on HMG-CoA reductase activity in hamster adrenals. PMID- 6273680 TI - Biosynthesis of beta-endorphin by the neurointermediate lobes from rats treated with morphine or alcohol. PMID- 6273681 TI - alpha 2-Adrenergic receptors in platelet membranes of depressed patients: no change in number of 3H-yohimbine affinity. PMID- 6273682 TI - Detection of erythrocyte membrane protein alterations in hereditary spherocytosis through the use of thermal stress: a spin label study. PMID- 6273683 TI - [Anthropometric nutritional indices and food intake of a child population of South Ivory Coast (author's transl)]. AB - The present study was carried out in order to determine new biochemical or immunological parameters which would be used to make an early diagnosis of protein-caloric malnutrition. For the first phase of the research, based on antropometrics (circumference of the arm, head, weight) and on the quantitative evaluation of daily nutritional food intake, three groups of children between the ages of 1 and 3 were selected according to the following criteria: -- Properly nourished children, -- Malnourished children without physical signs of malnutrition, -- Malnourished children with malnutrition signs. The description of these three groups using the forementioned indicators is reported. This work performed on 652 infants shows the following results: -- Whatever the age, mid arm circumference and weight are significantly reduced in confirmed cases of malnutrition, but are not useful in detecting malnutrition at an early stage. -- Whatever the age, head circumference does not change with the nutritional status, but it is reduced when the malnutrition has set in. -- Whatever the age, the caloric ration (25% deficit) and the proteic ration (35% deficit) of a malnourished child without signs of malnutrition and those of a well nourished child are different. But those ration does not change with an increasing degree of malnutrition. PMID- 6273684 TI - [Thoughts on the postoperative after care by bronchial cancer]. PMID- 6273685 TI - [Experimental silicosis. II. Effects of high temperature of fibrogenic effect of quartz and quartz sands used in ceramics]. AB - Fibrogenic effects of siliceous ceramic raw materials (quartz and quartz sand), heated at 1200 degrees C for 72 hours (conditions of ceramic products burning) have been investigated. The evaluation of fibrogenic effects has been based on wet lungs weight and hydroxyproline content in rats' lungs after an intratracheal administration of 50 mg of the test dust, during 3 and 6 months' experiment. The obtained results indicate that high temperature affects the polymorphous transformation of silica. The transformation varies, e.g. crystabillite has been found in quartz, whereas in quartz sands, quartz has been found apart from crystaballite. In the Isere quartz from Nowosolna crystalline silica content has been decreased, as compared to unheated samples. Heating does not reduce fibrogenic effects of some samples e.g. quartz sand from Biala Gora), or the activity may be increased (e.g. quartz sand from Winiary), or is not changed (quartz from Austria). The tendency of changes in biological aggressiveness of siliceous dusts subjected to high temperature cannot be foreseen, as those changes depend on different factos, among others on the content of various impurities. PMID- 6273686 TI - Comparative studies on fatty acid synthesis, glycogen metabolism, and gluconeogenesis by hepatocytes isolated from lean and obese Zucker rats. AB - Hepatocytes isolated from genetically obese female Zucker rats and lean female Zucker rats were compared. Hepatocytes from fed obese rats exhibited greater rates of fatty acid synthesis, more extensive accumulation of lactate and pyruvate from their glycogen stores, increased rates of net glucose utilization but produced less ketone bodies from exogenous fatty acids and had lower citrate levels than hepatocytes from lean rats. Lipogenesis was not as sensitive to dibutyryl cyclic AMP (DBcAMP) inhibition in hepatocytes from obese rats but glycogenolysis was stimulated to the same extent by this nucleotide in both preparations. Ketogenesis was less sensitive to stimulation by DBcAMP in hepatocytes from obese rats. A difference in sensitivity of lipogenesis to DBcAMP was not found when lactate plus pyruvate was added to the incubation medium, suggesting that a greater rate of glycolysis by hepatocytes from obese rats accounts for their relative insensitivity to DBcAMP. Citrate levels were elevated by DBcAMP to a greater extent in hepatocytes from obese rats. Hepatocytes prepared from lean rats starved for 48 hr were glycogen depleted and lacked significant capacity for lipogenesis and glycogen synthesis. In contrast, hepatocytes isolated from starved obese rats retained considerable amounts of liver glycogen and exhibited detectable rates of lipogenesis and glycogen synthesis. Hepatocytes prepared from starved lean rats gave faster apparent rates of lactate gluconeogenesis than hepatocytes prepared from starved obese rats. Thus, hepatocytes prepared from obese Zucker rats are more glycogenic, glycolytic, and lipogenic but less ketogenic and glucogenic than hepatocytes prepared from lean rats. PMID- 6273687 TI - DEAE-silica gel and DEAE-controlled porous glass as ion exchangers for the isolation of glycolipids. PMID- 6273688 TI - Assay for phospholipase C. PMID- 6273689 TI - Assay of enzymes of lipid metabolism with colored and fluorescent derivatives of natural lipids. PMID- 6273690 TI - Preparative isolation of lecithin. PMID- 6273691 TI - Extraction and purification of polyphosphoinositides. PMID- 6273692 TI - Perifusion of fat cells. PMID- 6273693 TI - Separation of methyl esters of fatty acids by gas chromatography on capillary columns, including the separation of deuterated from nondeuterated fatty acids. PMID- 6273694 TI - Effect of immune heterozygous spleen cell transfer on resistance to mouse hepatitis virus infection in nude mice. AB - The role of cell mediated immune response to mouse hepatitis virus (MHV) infection in mice was studied by transferring spleen cells from immune heterozygous littermates (nu/+). A suppressive effect on viral growth was seen in infected nude (nu/nu) mice, whereas immune nu/+ serum transfer had no effect. The protective effect of immune nu/+ spleen cells was significantly reduced by treatment with anti-theta serum plus complement but not with anti-Ig serum. In infected nu/nu mice which received transfers of immune nu/+ cells, neutralizing antibody appeared although the titer was not high enough to protect nu/nu mice from fatal infection. Histopathologically, lymphocyte infiltration in hepatic lesions was evident in infected nu/nu mice with nu/+ cell transfer, while it was slight without nu/+ cell transfer. PMID- 6273695 TI - Comparison of three electron microscopy techniques for the detection of human rotaviruses. AB - Rotavirus detection by direct electron microscopy was compared with direct and indirect immune electron microscopy techniques. The latter two approaches permitted the enumeration of 25 and 103 times more rotaviruses respectively, than direct electron microscopy. Also, 70% and 90% of the virus particles were aggregated by direct and indirect immune electron microscopy techniques respectively, thus facilitating their detection. PMID- 6273696 TI - Sequential passages of human rotavirus in MA-104 cells. AB - Starting with a small amount of diarrheal feces containing human rotavirus (HRV), we succeeded in propagation of the virus using the roller culture technique with MA-104 cells. Furthermore, we made a successful adaptation of HRV to a stationary culture and developed a plaque assay for the cell culture-adapted viruses. The 3 culture-adapted virus isolates, KU, YO, and 44 produced plaques (about 0.5-1.0 mm in diameter) under the overlay medium consisting of 0.6% purified agar, 3 micrograms of acetyl trypsin/ml and 50 micrograms of DEAE-dextran/ml. Subsequent plaque purification resulted in the formation of clear, larger plaques. It was shown from the results of cross neutralization tests using the fluorescent focus reduction method that the three culture-adapted HRV isolates were clearly different antigenically from ovine rotavirus (NCDV) and, further, that a noticeable difference in antigenicity also existed among the HRV isolates. PMID- 6273697 TI - A micro-neutralization system for detection of s-CRN antibody to herpes simplex virus. AB - It was learned that the ordinary micro-neutralization system with herpes simplex virus (HSV) gave a composite result of the initial neutralization and the effect of antibody on subsequent growth of unneutralized virus. In the case of slow reacting complement-requiring neutralizing (s-CRN) antibody, which was detected by incubating virus-serum mixtures at 4 C for 3 days before addition of complement, the titer obtained was lower than expected from the result of the plaque reduction test. This was thought ascribable to its low ability to prevent viral breakthrough caused by growth of unneutralized virus. This was overcome by adding an appropriate amount of hyperimmune antibody at 3 hr after addition of cells. The endpoint of s-CRN antibody so determined was but slightly lower than that obtained by the plaque reduction test. Early (1-week) rabbit sera, which were negative in the ordinary micro-neutralization test, titered 1:2,560 to 1:5,120 when tested by this method. When the 3-day sensitization in the cold was substituted by 5-hr incubation at 37 C, the titer obtained was 2 to 4-fold lower; in this case, however, the whole process could be finished within 3 days. Also, s CRN antibody reactive with type 2 HSV in homologous and heterologous sera could be detected by the same method using type 1 hyperimmune serum as the additional antibody. PMID- 6273698 TI - T cell growth factor from human splenic cell cultures: conditions for its production, and its utilization for maintenance of cytotoxic T cell lines. AB - We prepared the T cell growth factor (TCGF) from human spleen cell cultures stimulated with phytohemagglutinin (PHA). Various cell culture conditions and agents supporting the active TCGF production of the spleen cells were examined. The highest TCGF activity was obtained in the supernatants under the conditions that 2 x 10(6)/ml spleen cells were stimulated with PHA for 48 hr. Production of TCGF from spleen cells depended markedly on their individual sources. Addition of indomethacin to the culture or irradiation of the responding spleen cells increased TCGF activity in the supernatant of the culture. Further, addition of irradiated cells of an Epstein-Barr virus (EBV)-transformed lymphoblastoid cell line (LCL) to spleen cell cultures stimulated with PHA greatly enhanced TCGF production. Human splenic TCGF facilitated the establishment of human cytotoxic T cell (Tc) lines specific for EBV-transformed LCL cells when those Tc line cells were stimulated periodically with irradiated autologous LCL cells but not with the other two types (K-562 or Molt-4) of cells. Allogeneic LCL stimulators allowed the Tc line cells to proliferate. However, Tc line cells cocultured once with allogeneic LCL stimulators no longer exhibited EBV-specificity in their cytotoxicities. PMID- 6273699 TI - Isolation of Co1E1 mutants with thermosensitive control of colicin E1 production. PMID- 6273700 TI - Neutralizing patterns of anti-bovine rotavirus (Lincoln) serum against cytopathic bovine rotaviruses isolated from calves in Japan. PMID- 6273701 TI - Antigenic analysis of polioviruses isolated from a child with agammaglobulinemia and paralytic poliomyelitis after Sabin vaccine administration. AB - A 3-year-old boy with agammaglobulinemia developed paralytic poliomyelitis on day 553 after being fed poliovaccine. Non-vaccine-like type 2 polioviruses were isolated from 22 stools obtained within 684 days after the onset of illness. Antigenic variations were observed among these viruses. The non-vaccine-like virus isolated 1 week after the onset of paralysis differed in virulence from the Sabin type 2 vaccine strain in the neurovirulence test in monkeys, and did not have the same antigenic character as the wild virulent strains. Another virus isolated on day 348 before the onset of illness was also classified as non vaccine-like. However, the Sabin type 2 strain was shown to be homologous with this strain by the McBride test. Some Sabin-like particles were found in this stock virus. We may conclude that the non-vaccine-like virus isolates were derived from Sabin vaccine by antigenic variation that occurred during long-term multiplication in the intestinal tract. PMID- 6273702 TI - Characterization of two inducible bacteriophages, alpha 1 and alpha 2, isolated from Clostridium botulinum type A 190L and their deoxyribonucleic acids. AB - Two inducible bacteriophages, alpha 1 and alpha 2, isolated from Clostridium botulinum type A strain 190L and their deoxyribonucleic acids (DNAs) were purified and characterized. Phage alpha 1, which is unable to form plaques on any strain of C. botulinum, was produced in large quantities after treatment with mitomycin C (MC), whereas phage alpha 2, which was induced in much lower quantities than phage alpha 1, propagated in cultures of type A strain Hall. The phage DNAs were exclusively synthesized after induction with MC. Alpha 1 and alpha 2 DNAs had sedimentation coefficients of 34.0 and 30.6 S, corresponding to molecular weights of 31.9 x 10(6) and 23.5 x 10(6), respectively. The buoyant density in CsC1 was 1.682 g/cm3 for alpha 1 DNA and 1.680 g/cm3 for alpha 2 DNA. Based on thermal denaturation characteristics, the genomes of both phages were shown to be double-stranded DNAs. Agarose gel electrophoretic profiles of the phage DNAs digested with restriction endonuclease EcoRI revealed nine fragments for alpha 1 DNA and six fragments for alpha 2 DNA. The molecular weights of the phage DNAs as determined by restriction enzyme analysis were 30.55 x 10(6) for alpha 1 DNA and 25.83 x 10(6) for alpha 2 DNA. Nontoxigenic mutants obtained from strain 190L could, like the toxigenic parent strain, produce the two phages after treatment with MC. Lysogenic conversion to toxigenicity by phage alpha 2 was not observed with the nontoxigenic mutants. It seems likely that there is no relationship between either phage genome and the toxigenicity of C. botulinum type A. PMID- 6273703 TI - Lamellar structures found in cells infected with herpes simplex virus type 1. PMID- 6273704 TI - [Electron microscopic study of Bacillus polymyxa in the developmental process and antibiotic formation]. AB - The purpose of this work was to study growth of Bacillus polymyxa 153, biosynthesis of polymyxin by it, the formation of spores, and the fine structure of the cells. The antibiotic content was low in the cells and in the cultural broth during the first hours of growth and sharply increased later under the conditions of the medium ensuring the "vegetative" type of cultural growth. The cell wall reorganized, a periplasmic space appeared, and the membranous and mesosomal apparatus became more complicated in the cells actively synthesizing the antibiotic. The antibiotic content was higher in the cells belonging to the "spore" type of growth at the beginning of mass spore formation. Further cell development involved endospore differentiation; a complicated membranous apparatus did not appear, and biosynthesis of the antibiotic stopped. PMID- 6273705 TI - [Pleural metastases can simulate a mesothelioma]. AB - We are reporting about an inoperable cell carcinoma of the stomach, which exclusively caused thoracal pains due to metastases into a pleural thickening. First this symptomatology with thoracal pains and recurring pleural effusions was thought to be a pleural mesothelioma. Only the immediate histological examination of the tissue showed the origin of the tumor. Although cancer of stomach appears very commonly, this tumor is often diagnosed too late and in most cases it is already inoperable. Even nowadays more than 75% of the patients with a cancer of the stomach die of their metastases, although diagnosis and therapy have been developed. - Pleural mesotheliomas are rare; nevertheless the possibility of a mesothelioma in connection with all pleural effusions of a vague genesis should be considered. The exposure to asbestos strengthens the suspicion that this tumor is existing. In connection with a pleural mesothelioma an indurative pleurisy, a primary cancer of the lung and - like in our case - pleural metastases of a primary cancer of another site have to be differentiated. PMID- 6273706 TI - [Rheumatoid arthritis in children. Bases for a rational treatment]. PMID- 6273707 TI - [Serological markers correlated with hepatitis B virus. Study made predominantly in chronic hepatitis in children]. PMID- 6273708 TI - Tc-99m pyrophosphate myocardial imaging in acute myocardial infarction: value and limitations. PMID- 6273709 TI - The GABA postsynaptic membrane receptor-ionophore complex. Site of action of convulsant and anticonvulsant drugs. AB - The function of the inhibitory neurotransmitter, gamma-aminobutyric acid (GABA), has been implicated in the mode of action of many drugs which excite or depress the central nervous system. Many convulsant agents appear to block GABA action whereas anticonvulsants enhance GABA action. Some of these drug effects involve altered GABA-mediated synaptic transmission at the level of GABA biosynthesis, release from nerve endings, uptake into cells, and metabolic degradation. A greater number of agents of diverse classes appear to affect GABA action at the postsynaptic membrane, as determined from both electrophysiological and biochemical studies. The recently developed in vitro radioactive receptor binding assays have led to a wealth of new information about GABA action and its alteration by drugs. GABA inhibitory transmission involves the regulation, by GABA binding to its receptor site, of chloride ion channels. In this GABA receptor-ionophore system, other drug receptor sites, one for benzodiazepines and one for barbiturates/picrotoxinin (and related agents) appear to form a multicomponent complex. In this complex, the drugs binding to any of the three receptor categories are visualized to have an effect on GABA-associated chloride channel regulation. Available evidence suggests that the complex mediates many of the actions of numerous excitatory and depressant drugs showing a variety of pharmacological effects. PMID- 6273711 TI - Studies on structure and function of chromatin. AB - This article covers research work in this laboratory on the structure and function of chromatin. Early studies have led to discovery of skeletal fibrils (nucleonemas) within the nuclei and showed the specific role of histone H1 in chromatin condensation and in restriction of transcription. More recently with the aid of a novel DNP electrophoresis technique the relation of histone H1 and non-histone proteins to nucleosomes was studied. Three types of mononucleosomes and a number of subnucleosomes were identified in chromatin digests. The complexes of certain HMG proteins with short DNA segments were isolated and found to originate frm transcriptionally active chromatin. Different forms of SV40 minichromosome were characterized. A method for the analysis of nucleosome distribution along the DNA sequence was elaborated and used to show non-random (phased) location of nucleosomes on SV40 DNA. The attachment of DNA to skeletal elements of interphase nuclei and metaphase chromosomes was shown to be a non random, probably sequence-specific process. PMID- 6273712 TI - Superpotency and superaffinity phenomena in the stimulation of steroidogenesis in adrenocortical cells by adrenocorticotropin-tobacco mosaic virus conjugates. AB - Adrenocorticotropin-(1-24)-tetracosapeptide was covalently attached to tobacco mosaic virus in two different manners: (i) through a handle near the C-terminus on tyrosine-(23) and (ii) through a handle at the N-terminus on serine-(1). Compounds of type (i) with their N-terminal message sequence freely exposed on the virion surface were considerably more potent for stimulating steroidogenesis in isolated adrenocortical cells than those of type (ii) with a more congested message. Conjugates with 50 or less hormone molecules per virion were less potent per peptide unit than the "free" handle-substituted hormones, whereas conjugates with 150 ACTH units exhibited superpotency effects. Superpotency disappeared when the substituted virions were disaggregated into (substituted) capsomers, suggesting influences of hormone clustering and virion geometry on biological activity. Superpotent stimulation was irreversible under conditions that immediately inhibited steroidogenesis by ACTH (dilution, addition of a peptide antagonist). Thus, superpotency might be caused by superaffinity arising from a slow rate of dissociation of the conjugates from the target cell receptors. The reason for the slow dissociation rate is still unclear: possible explanations include cooperative affinity, rapid internalization of the conjugate-receptor complexes, or decreased rates of peptide degradation at the receptor site. PMID- 6273713 TI - The soluble, cyclic AMP-stimulated protein kinase catalyses the phosphorylation of different membrane proteins from those which are phosphorylated by the membrane bound enzyme. AB - Soluble, cyclic AMP stimulated protein kinase (type a1 protein kinase) stimulated the phosphorylation of membrane proteins in preparations from a wide variety of tissues. This kinase does not, however, appear to catalyse the phosphorylation of the same membrane proteins which are phosphorylated by the intrinsic protein kinase activity of the preparations. This was demonstrated in two ways: 1. If preparations of membrane fragments are incubated with [gamma 32P] ATP in the absence of type a1 protein kinase, until all membrane bound protein acceptor sites are saturated with phosphate addition of type a1 protein kinase causes the incorporation of more phosphate into the membrane proteins. 2. The nature of the membrane proteins phosphorylated by type a1 protein kinase and the intrinsic protein kinase of the preparation was compared by SDS gel electrophoresis. Membrane preparations were phosphorylated with [gamma 32P] ATP in the absence of added type a1 protein kinase and with [gamma 33P] ATP in the presence of the enzyme. Aliquots of the two samples were then mixed, separated by SDS gel electrophoresis, and the distribution of radioactivity measured. A comparison of the distribution of protein bound 33P and 32P made it quite clear that type a1 protein kinase catalysed the phosphorylation of different membrane proteins from those which are phosphorylated by the intrinsic protein kinase activity of the preparations. PMID- 6273710 TI - Cellular compartments of GABA in brain and their relationship to anticonvulsant activity. PMID- 6273714 TI - Activation of Cl, the first component of complement, the generation of Clr-Cls and Cl- inactivator complexes in normal serum by heparin-affinity chromatography. PMID- 6273715 TI - Stimulatory effect of latex and zymosan particles on cyclic adenosine 3',5' monophosphate content in human granulocytes. PMID- 6273716 TI - The recognition of acute myocardial infarction occurring during the immediate postoperative period. PMID- 6273717 TI - [Fast serologic diagnosis in virus diseases]. PMID- 6273718 TI - [Practical sports medicine. Part 12. After care of injuries]. PMID- 6273719 TI - [Medical sociology and community-centered care of psychic patients. Methodical considerations of a concrete research project (author's transl)]. AB - The need, problems and possible approaches to sociological investigation of community-centered care of psychological patients is the theme of this contribution. It will at the same time sketch out, even if only in very rough outline, a direction for present day medical sociologic research which has its origin in psychiatric research but which, under the present sociopolitical and care relations, it is necessary to submit to the extramedical field of rehabilitation and aftercare for psychic patients at communal and regional levels according to the importance. PMID- 6273721 TI - Late changes in the motor unit after acute poliomyelitis. AB - Patients who have suffered an acute attack of poliomyelitis may develop a clinically evident progression of weakness later in life. A syndrome known as forme fruste amyotrophic lateral sclerosis occurs in severely affected older polio patients. Neither process is well understood. Ten subjects who had had polio at least 22 years prior to examination and had no complaints of rapid progression of weakness were studied. Single-fiber recordings were made from the extensor digitorum communis or the tibialis anterior muscle. Fiber densities were increased in all patients. Eight of 10 patients showed marked abnormalities in jitter in more than 50% of the recordings examined. There was a significant association between the percentage of recordings with abnormal jitter and the time since the attack of polio. Blocking was seen in more than 30% of the recordings examined in 6 of 10 patients. There was a significant association between the percentage of recordings demonstrating blocking and both the chronological age and the number of years since the attack of polio. These abnormalities may represent disintegration with aging in the reinnervated motor units. PMID- 6273720 TI - Biochemical functioning of mitochondria in normal and denervated mammalian skeletal muscle. AB - Mitochondrial preparations derived from denervated rat skeletal muscle and paired control muscle are characterized with respect to their oxidative and phosphorylative capacities. Our data indicate that there is an impairment within the first 2 energy coupling regions of the respiratory chain and within the ATPase complex itself. PMID- 6273722 TI - Herpesvirus-induced antigens in squamous cell carcinoma in situ of the vulva. PMID- 6273723 TI - Exercise and the endogenous opioids. PMID- 6273724 TI - Ectopic ACTH production with autoantibody formation in a patient with acute myeloblastic leukemia. PMID- 6273725 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. 53-1981. Persistence of Cushing's syndrome after hypophysectomy. PMID- 6273726 TI - Isolation of enterovirus 70 from patients with acute hemorrhagic conjunctivitis in Key West, Florida. PMID- 6273727 TI - Pyrophosphate imaging for infarct sizing. PMID- 6273728 TI - EBV molecular epidemiology: lack of strain role in disease manifestation. PMID- 6273729 TI - Tracing outbreaks of foot-and-mouth disease. PMID- 6273730 TI - Single channel recordings of Ca2+-activated K+ currents in rat muscle cell culture. PMID- 6273731 TI - Biochemical identification of viruses causing the 1981 outbreaks of foot and mouth disease in the UK. PMID- 6273732 TI - Production in B. subtilis of hepatitis B core antigen and a major antigen of foot and mouth disease virus. PMID- 6273733 TI - SV40 recombinant molecules express the gene encoding p21 transforming protein of Harvey murine sarcoma virus. PMID- 6273734 TI - Protein phosphatase-1 is involved in Xenopus oocyte maturation. PMID- 6273735 TI - Opposing roles for D-1 and D-2 dopamine receptors in efflux of cyclic AMP from rat neostriatum. PMID- 6273736 TI - Single channel potassium currents of the anomalous rectifier. PMID- 6273737 TI - Variant insertion element IS1 generates 8-base pair duplications of the target sequence. PMID- 6273738 TI - A potpourri of plucked patches. PMID- 6273739 TI - Receptors, channels and whole-body NMR in Mexico. PMID- 6273740 TI - Regulation of epithelial tight junction permeability by cyclic AMP. PMID- 6273741 TI - Identification of antigenic determinants unique to the surfaces of cells transformed by Epstein-Barr virus. PMID- 6273743 TI - Fluctuations in the microsecond time range of the current through single acetylcholine receptor ion channels. PMID- 6273742 TI - Multiple conductance states of single acetylcholine receptor channels in embryonic muscle cells. PMID- 6273744 TI - GABA reduces binding of 3H-methyl beta-carboline-3-carboxylate to brain benzodiazepine receptors. PMID- 6273745 TI - E. coli uvrB protein binds to DNA in the presence of uvrA protein. PMID- 6273746 TI - Protein-lipid interactions: do the spectroscopists now agree? PMID- 6273747 TI - Clustered arrangement of immunoglobulin lambda constant region genes in man. AB - The immunoglobulin lambda light chain locus of man contains six lambda-like genes arranged tandemly on a 50-kilobase segment of chromosomal DNA. THe sequences of three of these genes correspond to three known non-allelic lambda chain isotypes: Mcg, Ke-Oz- and Ke-Oz+. They surround a highly polymorphic and evidently unstable region that is repeatedly deleted when cloned in Escherichia coli. Three additional, but as yet unlinked, lambda-like sequences have also been cloned, suggesting that the lambda genes form an unexpectedly large family within the human genome. PMID- 6273748 TI - N alpha-acetylation is linked to alpha-MSH release from pars intermedia of the amphibian pituitary gland. PMID- 6273749 TI - Specific integration of REV proviruses in avian bursal lymphomas. PMID- 6273750 TI - Autoradiographic localization of GABAB receptors in rat cerebellum. PMID- 6273751 TI - Neurotensin receptors are located on dopamine-containing neurones in rat midbrain. PMID- 6273752 TI - Action of glutamate and aspartate analogues on rod horizontal and bipolar cells. PMID- 6273753 TI - DNA supercoiling by DNA gyrase. PMID- 6273754 TI - Site-specific recombination. PMID- 6273755 TI - Presence and physiological role of presynaptic inhibitory alpha 2-adrenoreceptors in guinea pig atria. PMID- 6273756 TI - [Experiences with intravenously administered acyclovir in severe herpesvirus infections]. PMID- 6273757 TI - Acidosis and renal calcium excretion in experimental chronic renal failure. AB - In renal failure, absolute calcium excretion is low, but fractional excretion (FE) of filtered load is increased. In order to determine the role of metabolic acidosis in contributing to increased FECa, we have studied thyroparathyroidectomized dogs in a control phase and following the induction of chronic renal failure, both during spontaneous metabolic acidosis and after correction with NaHCO3. FECa was 3.7% in controls and increased to 13.7% in azotemic acidotic dogs (p less than 0.01). After correction of acidosis FENa was not significantly changed, but FECa fell significantly, to 8.1% (p less than 0.01), while glomerular filtration rate, plasma calcium and filtered calcium load were unchanged. Thus although FECa is increased in nonacidotic azotemic dogs, acidosis further enhances calcium excretion by inhibiting renal tubular calcium reabsorption. These effects of metabolic acidosis may contribute to hypocalcemia and bone disease in azotemia. PMID- 6273758 TI - Rat dopaminergic function in the retina during aging. AB - Parameters of dopaminergic transmission were studied in the retina of mature (3-4 months) and aged (23-24 months) rats. In the retina of senescent rats were found significantly higher dihydroxyphenylacetic acid (DOPAC) levels and a higher number of (3H-)spiroperidol binding sites. We detected also an increase of (3H)- methionine-enkephalin binding sites. The changes in the density of (3H) spiroperidol and (3H)-Metenkephalin binding sites in the retina are opposite to those observed in the brain of aged rats. PMID- 6273759 TI - GTP-sensitive phosphorylation of proteins in a postmitochondrial supernatant from rat brainstem affected by ACTH1-24. AB - The effect of ACTH1-24 and cyclic nucleotides on the endogenous phosphorylation of proteins from a postmitochondrial supernatant from rat brainstem was investigated in the presence and absence of GTP. Phosphorylation and its modulation by these compounds were studied in vitro by incorporation of labeled phosphate from [gamma-32P]ATP added to the incubation mixture. Phosphoproteins were subsequently analyzed by autoradiography after one- and two-dimensional separation. Eight ACTH-sensitive phosphoproteins of molecular weights 75 (IEP 4.0), 67, 64, 50 (IEP 4.7), 47 (IEP 4.8), 38, 34, and 24K were found. The effects of ACTH on phosphorylation were mainly inhibitory, and the affected protein bands did not coincide with the phosphoproteins sensitive to cyclic AMP and cyclic GMP. Phosphorylation of those phosphoprotein bands and its ACTH sensitivity appeared to be highly sensitive to GTP. It is suggested that the activity of protein kinases involved in hormone-sensitive phosphorylation in a postmitochondrial rat brainstem fraction is regulated by GTP-dependent mechanisms. PMID- 6273760 TI - Further studies on mevalonate phosphorylation in neonatal chick brain. AB - Mevalonate phosphorylation was studied in neonatal chick brain. Formation of phosphorylated derivatives of mevalonic acid increased with the pH in the range assays (5.5-9.5). Phosphomevalonate kinase was completely inactivated after treatment at 50 degree C for 5-10 min, whereas mevalonate kinase was found to retain its activity under the same conditions. Exposure to 65 degree C for 5 min resulted in the inactivation of mevalonate kinase. Both mevalonate-activating enzymes from chick brain were located primarily in the soluble fraction. The amounts of phosphomevalonic acid and pyrophosphomevalonic acid did not show a significant diurnal variation to suggest the presence of a circadian rhythm in either kinase. Cholesterol feeding and fasting had no effect on mevalonate phosphorylation by neonatal chick brain. PMID- 6273761 TI - [Cerebrospinal fluid cAMP and cGMP in various peripheral and central nervous system diseases. Preliminary report]. AB - In a group of 51 patients the concentrations of cAMP and cGMP and their ratio were determined in the cerebrospinal fluid in certain neurological diseases. Statistically significant differences were found in the values of the concentrations of the above metabolites in selected groups of neurological diseases. Generally speaking, apart from a group of multiple sclerosis cases the values of cAMP concentration were higher in central nervous system diseases than in peripheral nervous system diseases. The highest values of the cAMP/cGMP ratio were observed in the group of patients with inflammatory conditions of the central nervous system. This method of cerebrospinal fluid examination may have an important role in the diagnosis of neurological diseases, particularly in multiple sclerosis. PMID- 6273762 TI - [Clinical trial of suloctidil (sulocton) for the treatment of migraine (preliminary report)]. AB - Suloctidil, a new vasoactive drug, shows certain properties which make it particularly suitable for using in migraine. The author administered Suloctidil in daily doses of 600 mg to 12 patients with migraine during 6 weeks. In 9 cases good results were obtained. Although further investigations are necessary, it seems already now that Suloctidil will enrich the armamentarium of antimigrainous drugs. PMID- 6273763 TI - A study of psychological and endocrine variables on 14 patients treated by chronical haemodialysis. AB - In 13 patients suffering from renal dysfunction and treated by chronic haemodialysis, the mnemic functions appeared within the limits of normality and there was no positive correlation between blood levels of neurophysines and free cortisol. On the other hand, a negative correlation was found between levels of neurophysine I and items 3 and 5 of the memory test PRM. This negative result indicates that in haemodialyzed patients a discrepancy may exist between blood levels of neurophysines and vasopressin release, or that there is no direct relationship between plasma and CSF concentrations of such hormones. PMID- 6273764 TI - [Opiate receptors and sleep. III. Effects of microinjections of DSIP (delta-sleep peptide) in the median thalamus, Periaqueductal gray matter and nucleus of the tractus solitarius of the rabbit (author's transl)]. AB - In the rabbit, microinjections of DSIP (delta-sleep peptide) at optimal doses in the median thalamus, periaqueductal gray matter (22.5 nmol), and nucleus of the tractus solitarius (15.0 nmol) produce slow-wave sleep with abundant recruiting spindles. As is seen with morphine, this effect is blocked by naloxone (160 micrograms intracerebrally; i.c.). At least in the rabbit, DSIP is probably a neurotransmitter or a neuromodulator of the bulbothalamic system inducing slow wave sleep and particularly recruiting spindles. It is likely to restore slow wave sleep after supraoptimal awakening i.c. microinjections of morphine. PMID- 6273765 TI - Sjogren syndrome: central nervous system manifestations. AB - We studied eight patients who had primary Sjogren syndrome and central nervous system (CNS) disorders that were not attributable to other causes. Focal cerebral deficits were observed in five patients. Aseptic meningoencephalitis was seen in five patients, recurrent in one. Spinal cord manifestations in three patients took several forms: acute transverse myelitis, chronic progressive myelopathy, and spinal subarachnoid hemorrhage. Precipitating antibodies to the Ro(SSA) cytoplasmic antigen were detected in the sera of seven of eight patients. This may be relevant to the pathogenesis of CNS disease in Sjogren syndrome, because there is a strong correlation between vasculitis and the presence of anti-Ro(SSA) antibodies in this connective tissue disorder. PMID- 6273766 TI - Giant axonal neuropathy: intermediate filament aggregates in cultured skin fibroblasts. AB - Giant axonal neuropathy is characterized by large masses of filaments in neurons and other cells. Coomassie blue staining of the detergent-resistant cytoskeleton of cultured fibroblasts from three patients revealed cytoplasmic masses with the characteristics of aggregates of intermediate (8- to 10-nm) filaments. These fibroblast abnormalities provide definitive evidence for a genetic etiology of this disease and may be useful for prenatal diagnosis and for investigations of pathogenesis. PMID- 6273767 TI - Prevalence of monoclonal protein in peripheral neuropathy. AB - We attempted to survey with serum protein electrophoresis all patients with peripheral neuropathy of unknown cause presenting at our institution within 1 year to determine the prevalence of monoclonal serum proteins in this group. Eighty-four percent of these patients were evaluated, and 10% of these had a monoclonal serum protein. This figure was statistically different from that obtained in two community studies in the literature. Because these patients often present difficult diagnostic problems, serum protein electrophoresis is necessary in the evaluation of cryptogenic neuropathies. PMID- 6273768 TI - Dopaminergic mechanism in generalized photosensitive epilepsy. AB - Apomorphine, a dopamine receptor agonist, blocked epileptic photosensitivity in patients with primary corticoreticular epilepsy. This effect was not modified by naloxone, a narcotic antagonist, suggesting that apomorphine acts on cerebral dopaminergic receptors. Apomorphine did not block spontaneous spike-and-wave discharges in patients with nonphotosensitive primary corticoreticular epilepsy. The different actions of apomorphine on spontaneous and photically induced spike and-wave activity suggest that there is a selective dopaminergic mechanism in human epileptic photosensitivity. PMID- 6273769 TI - Neuromuscular transmission in rheumatoid arthritis, with and without penicillamine treatment. AB - Electroneuromyographic studies were performed on patients with rheumatoid arthritis, some of whom were received penicillamine, to determine whether a subclinical defect of neuromuscular transmission existed. There were no significant differences between patients and controls with respect to nerve conduction studies or repetitive ulnar nerve stimulation. Four patients (three receiving penicillamine) demonstrated mild neurogenic changes distally on needle electromyography. Mean jitter was slightly higher for patients receiving penicillamine than in other patients or controls, but the differences were not significant. No significant correlations existed between of the studies and daily, cumulative, or average penicillamine dosage. A significant positive correlation (p less than 0.001) existed between jitter and duration of disease in patients receiving penicillamine. Results were consistent with the hypothesis that penicillamine predisposes certain individuals to develop myasthenia gravis rather than interfering directly with neuromuscular transmission. PMID- 6273770 TI - [Right hepatectomy. Personal considerations]. PMID- 6273771 TI - Alcoholic peripheral neuropathy. PMID- 6273772 TI - Attenuation of turkey meningo-encephalitis virus in BHK21 cells. AB - Turkey meningo-encephalitis virus was adapted to BHK21 cell culture. Cytopathic effects were characterized by rounding and detachment of cells within 48 hours. Attenuation was achieved by 41 successive passages in BHK21 cell cultures. Turkeys and Japanese quail (Coturnix coturnix japonica), kept under laboratory conditions and inoculated with the attenuated virus, did not develop symptoms of turkey meningo-encephalitis but reacted by the production of haemagglutination inhibition antibody. They resisted intracerebral challenge with pathogenic strains of turkey meningo-encephalitis virus. PMID- 6273773 TI - Identification of the serotype-specific and group-specific antigens of bluetongue virus. AB - The bluetongue virus (BTV) core particle contains 2 major polypeptides, P3 and P7, and is surrounded by an outer capsid layer that is composed of the 2 major polypeptides, P2 and P5. Analysis of the immune precipitates from soluble 14C labelled BTV polypeptides and hyper-immune rabbit and guinea-pig sera indicated that polypeptide P2 precipitates only with homologous BTV sera. This would indicate that P2 is the main determinant of serotype specificity. It was also found that in sheep infected with BTV the P2-precipitating antibodies in the serum correlate with the neutralizing antibody titres, whereas the appearance and subsequent decline of P7-precipitating antibodies correspond well with those of the complement fixing antibodies. This suggests that BTV group specificity, as measured by a complement fixation tests, is determined by the core protein P7. This result was supported by the observation that mouse ascitic fluid, which contains a high titre of BTV-specific complement fixing antibodies and a very low titre of neutralizing antibodies, contains almost exclusively antibodies that precipitate P7. PMID- 6273774 TI - A comparison of an australian bluetongue virus isolate (CSIRO 19) with other bluetongue virus serotypes by cross-hybridization and cross-immune precipitation. AB - No major differences in size were observed when both the double-stranded RNA and the polypeptides of the Australian bluetongue virus (BTV) isolate CSIRO 19 (BTV 20) were compared with those of other BTV serotypes such as BTV-10 and BTV-4. Minor capsid polypeptide P6 of both BTV-20 and BTV-4, which electrophoreses as a single band on continuous phosphate buffered gels, in separated into 2 distinct bands on discontinuous glycine-buffered gels. This was not the case with BTV-10. Cross-immune precipitation of BTV-20 with BTV-10, BTV-17, BTV-4 and BTV-3 indicated strong immunological cross-reaction of the group-specific antigen P7 of the different serotypes. There was also some cross-immune precipitation of the serotype-specific polypeptide P2 of BTV-20 and BTV-4. This result is in agreement with the observed cross neutralization of these 2 viruses. The main distinction between BTV-20 and the other BTV serotypes was observed in cross-hybridization experiments. The homology between the nucleic acid of BTV-20 and other BTV serotypes was less than 30%, whereas homology normally found between BTV serotypes is at least 70%. The hybridization products of the different BTV serotypes were analysed by electrophoresis and fluorography. Two main hybrid segments were observed in all heterologous hybridizations with BTV-20 as a compared with 7 hybrid segments in hybridizations between BTV-4 and BTV-10. In order to determine from which genome segment of BTV-20 these 2 hybrid segments were derived, the hybridizations were carried out with individually purified double-stranded RNA segments. These results indicate that the 2 segments of BTV 20 that show the largest homology to corresponding segments of a heterologous BTV serotype are No. 7 and 10. PMID- 6273775 TI - [Radioisotope studies in double distraction epiphyseolysis of the tibia]. PMID- 6273776 TI - [Leukocyte migration inhibition evoked by Epstein Barr virus antigen. I. Immune response in healthy donors]. PMID- 6273777 TI - [Surgical treatment of parotid neoplasms]. PMID- 6273778 TI - [Case of mixed tumor of the nasal vestibule]. PMID- 6273779 TI - Tumors of the blood vessels and of the mesothelial coverlayers. PMID- 6273780 TI - Organ culture in the study of human tumors with special reference to nephroblastoma. PMID- 6273782 TI - Variation on a theme by Fenner: the pathogenesis of chickenpox. PMID- 6273781 TI - [Histological identification of carcinoembryonic antigen in breast cancer. Use of an immunohistochemical technique in 71 cases of breast cancer (author's transl)]. PMID- 6273783 TI - [Somatotropin and corticotropin content of the blood in children and adolescents with diabetes mellitus]. PMID- 6273784 TI - Effects of some ion transport inhibitors on secretion and reabsorption in intact and perfused single human sweat glands. AB - The effects of three electrolyte transport inhibitors on the secretory and absorptive function of the human eccrine sweat gland were examined. Amiloride and furosemide at 10(-4) mol/l had no detectable effect on secretion while ouabain at 10(-4) mol/l completely inhibited secretion. In contrast, when amiloride, ouabain and furosemide were applied at 10(-4) mol/l to the luminal surface of the sweat duct by microperfusion, only amiloride blocked Na reabsorption. When the same inhibitors were added to the bathing solution at the same concentration, only ouabain inhibited Na reabsorption detectably. Present findings suggest that the secretory epithelium is permeable to solute and water movement while the duct epithelium is probably impermeable with respect to fluid movement across it. PMID- 6273785 TI - Measurement of hepatic blood flow in the rat using fractional clearance of indocyanine green and colloidal radiogold. AB - Two new methods are described to measure hepatic blood flow in the anaesthetized rat. These methods are based on the fractional clearance and extraction of indocyanine green, which is removed by hepatocytes, and of colloidal radiogold, which is removed by Kupffer cells. Hepatic blood flow was found to be 2.11 +/- 0.35 ml . min-1 . g liver-1 (mean +/- SD) and 2.01 +/- 0.31 ml . min-1 . g liver 1, respectively, with these two substances (P greater than 0.80). PMID- 6273786 TI - [Polymorphism of hemoglobins D in Ivory Coast: Hb Korle Bu (beta 73 (E17) Asp leads to Asn), Hb Avicenna (beta 47 (CD6) Asp leads to Ala) and Hb Cocody (beta 21 (B3) Asp leads to Asn) (author's transl)]. AB - A recent screening performed at the CHU Cocody in Abidjan (Ivory Coast) revealed six cases of hemoglobin "D". The use of a rapid strategy based on isoelectric focusing studies, finger-printing, reverse phase high performance liquid chromatography and solid phase microsequence technique led to the characterization of three abdominal hemoglobins. The first variant corresponded to the Hb Korle Bu (beta 73 (E17) Asp leads to Asn) (pI = 7.210) which is common in this country. The second, Hb Avicenna (beta 47 (CD6) Asp leads to Ala) (pI = 7.225) has not yet been reported in Africa. The last one was a new variant, Hb Cocody (beta 21 (B3) Asp leads to Asn) (pI = 7.205). A large number of hemoglobin "D" have been detected in European, American negroes, Amerindian or Asiatic populations, but only few observations were reported in Africa. Large screenings and discriminative methodologists must provide information on the polymorphism of hemoglobin D in the African population. PMID- 6273787 TI - Restriction endonuclease EcaI from Enterobacter cloacae. AB - Restriction endonuclease EcaI obtained from Enterobacter cloacae DSM30056 recognizes the group of heptanucleotide palindromes 5'-G[unk]G-T-N-A-C-C-3', and on cleavage (arrow) produces fragments with 5'-terminal pentanucleotide extensions. It is identical in specificity with restriction endonuclease BstEII from Bacillus stearothermophilus ET. PMID- 6273788 TI - ClaI. a new restriction endonuclease from Caryophanon latum L. AB - From Caryophanon latum L site specific restriction endonuclease (ClaI) has been purified, which recognises tha DNA hexanucleotide palindrome 5'-A-T-C-G-A-T-3'. Staggered cleavage generates DNA restriction fragments with 5'-terminal pCG extensions. A CLaI map of bacteriophage lambda has been determined, which indicates cleavage inhibition due to adenine methylation at over lapping ClaI GATC recognition sequences. Plasmid pBR322 is cut only once, in the tetracycline promoter region, and can, therefore, be used as a vector system for cloning fragments derived from ClaI digestions, and in addition for fragments generated by TaqI, HpaII, and several other enzymes. PMID- 6273789 TI - There are approximately 20 actin gene in the human genome. AB - By three different lines of evidence there are approximately 20 copies of actin genes in the human genome. Firstly, the rate of hybridisation of a mouse actin probe to human DNA indicates that there are a minimum of 20 complementary copies of the actin sequence per genome. Secondly, this probe hybridises to 17-20 bands in Southern blots of restriction enzyme digests of total human DNA. Most of these bands hybridise with both 3' and 5' fragments of the cDNA and are therefore likely to contain the entire gene sequence. Thirdly, we have picked 12 actin recombinants from a genomic library, and at the level of restriction enzymes mapping these represent nine different genes. Probability calculations indicate that these recombinants were picked from a pool of at least 20 different genes. PMID- 6273790 TI - Structural requirements of polynucleotides for the activation of (2' - 5')An polymerase and protein kinase. AB - Two enzymatic pathways are involved in the inhibitory effects of double-stranded (ds)RNA on protein synthesis in cell extracts derived from interferon-treated human fibroblasts or HeLa cells, an oligonucleotide polymerase that synthesizes (2'-5')An from ATP and a protein kinase that phosphorylates the alpha subunit of initiation factor eIF-2 as well as a polypeptide of Mr = 72,000. We have now evaluated the activation of both the (2'-5')An polymerase and protein kinase by a large variety of polynucleotides, triple-stranded and synthetic dsRNAs, homopolymers, alternating copolymers, triple-stranded polymers, purine-purine duplexes and purine-pyrimidine duplexes with modifications at either the pyrimidine or ribose moieties. All these polynucleotides have been the subject of previous interferon induction studies. Some polynucleotides, i.e. (I)n.(C)n and mycophage dsRNA, which have been recognized as excellent interferon inducers, were also potent activators of both (2'-5')An polymerase and protein kinase, whereas non-inducers such as (A)n. (X)n and (A)n. (br5U)n did not activate either the kinase or the polymerase. However, some polymers like (I)n.(br5C)n, (difl)n(C)n and (dIcl)n (C)n, while potent interferon inducers and kinase activators, behaved poorly as activators of the (2'-5')An polymerase. Other polymers, i.e. (dAfl)n (U)n and (A)n.(U)nl (I)n, that do not induce interferon, activated the kinase but not the polymerase. Finally, (I)n (s2c)n, a relatively potent interferon inducer, did not activate either kinase or polymerase. These findings indicate that there is no simple relationship between the interferon inducing ability of dsRNAs and their stimulating effects on (2'-5')An polymerase and protein kinase activity. PMID- 6273791 TI - The endogenous proviral mouse mammary tumor virus genes of the GR mouse are not identical and only one corresponds to the exogenous virus. AB - The endogenous proviral copies of mouse mammary tumor virus (MMTV) were selected from a gene library of GR mouse DNA. We obtained five different lambda. MMTV recombinant clones. Four of them correspond to the 3' Eco RI fragments of the endogenous proviruses an one comprises an intact MMTV provirus with 2 to 3 kb of flanking mouse genomic DNA. Heteroduplex formation followed by S1 digestion under stringent conditions shows that there is nucleotide sequence heterology among the cloned endogenous proviral copies. Only one endogenous proviral copy, associated with the mtv-2 locus, was found to be totally homologous to the exogenous proviral DNA. PMID- 6273792 TI - Nucleotide sequences at the boundaries between gene and insertion regions in the rDNA of Drosophilia melanogaster. AB - Ribosomal RNA genes interrupted by type 1 insertions of 1 kb and 0.5 kb have been sequenced through the insertion region and compared with an uninterrupted gene. The 0.5 kb insertion is flanked by a duplication of a 14 bp segment that is present once in the uninterrupted gene; the 1 kb insertion is flanked by a duplication of 11 of these 14 bp. Short insertions are identical in their entire length to downstream regions of long insertions. No internal repeats occur in the insertion. The presence of target site duplications suggests that type 1 insertions arose by the introduction of transposable elements into rDNA. Short sequence homologies between the upstream ends of the insertions and the 28S' boundaries of the rRNA coding region suggest that short type 1 insertions may have arisen by recombination from longer insertions. We have sequenced both boundaries of two molecules containing type 2 insertions and the upstream boundary of a third; the points of interruption at the upstream boundary (28S' site) differ from each other in steps of 2 bp. Between the boundary in the 0.5 kb type 1 insertion and the type 2 boundaries there are distances of 74, 76, and 78 bp. At the downstream boundary (28S'' site) the two sequenced type 2 insertions are identical. The rRNA coding region of one molecule extends across the insertion without deletion or duplication, but a 2 bp deletion in the RNA coding region is present in the second molecule. Stretches of 13 or 22 adenine residues occur at the downstream (28S'') end of the two type 2 insertions. PMID- 6273793 TI - Ribosomal protein genes rp 39(10 - 78), rp 39(11 - 40), rp 51, and rp 52 are not contiguous to other ribosomal protein genes in the Saccharomyces cerevisiae genome. AB - A library of recombinant phage containing EcoR1 fragments of Saccharomyces cerevisiae DNA has been constructed. This library was screened with four different recombinant plasmids, each containing a different yeast ribosomal protein gene, in order to isolate chromosomal fragments extending in both directions from these genes. These chromosomal fragments were assayed for the presence of additional ribosomal protein genes by hybridization selection and cell free translation, and none were found. These four regions are not closely linked to each other, since DNA from one domain does not cross hybridize with DNA from any of the other three, except for the sequences within the homologous ribosomal protein 39 gene pair. Northern blots demonstrate that although the concentrations of ribosomal protein mRNAs are diminished significantly in a strain containing the ts mutation rna2, transcripts from genes in these flanking segments are relatively unaffected. PMID- 6273794 TI - DNA fragments associated with chromosome scaffolds. AB - Following extensive digestion of HeLa metaphase chromosomes with either Hae III endonuclease or micrococcal nuclease, nonhistone protein scaffolds may be isolated. Scaffolds isolated after Hae III digestion have about 1.5% of the chromosomal DNA attached to them. This DNA is heterogeneous in size, ranging from about 0.2 to 20 kbp. It can be cleaved with either Eco RI or Hae III - Eco RI, producing a series of repeated fragments, of which the most abundant is 1.7 kbp in length. The 1.7-kdp fragment is tandemly repeated and is enriched (about 50 fold) in the scaffold-associated DNA. It is located primarily on human chromosome 1 and is probably a component of human satellites II and III. Scaffolds isolated after micrococcal nuclease digestion have about 0.1% of chromosomal DNA attached. This DNA is present in two size classes - fragments larger than 10 kbp and fragments approximately 0.2 kbp long. Restriction enzyme digestion of this DNA gives no prominent repeated fragments. Its reassociation kinetics are similar to those of total DNA, indicating that it is not enriched in either highly repetitive or middle repetitive sequences. PMID- 6273796 TI - Duplicated rDNA sequences of variable lengths flanking the short type I insertions in the rDNA of Drosophila melanogaster. AB - We describe cloned segments of rDNA that contain short type I insertions of differing lengths. These insertions represent a coterminal subset of sequences from the right hand side of the major 5kb type I insertion. Three of these shorter insertions are flanked on both sides by a short sequence present as a single copy in uninterrupted rDNA units. The duplicated segment is 7, 14 and 15 nucleotides in the different clones. In this respect, the insertions differ from the 5kb type I insertion, where the corresponding sequence is found only at the right hand junction and where at the left hand side there is a deletion of 9 nucleotides of rDNA (Roiha et al.,1981). One clone is unusual in that it contains two type I insertions, one of which is flanked by a 14 nucleotide repeat. The left hand junction of the second insertion occurs 380 nucleotides downstream in the rDNA unit from the first. It has an identical right hand junction to the other elements and the 380 nucleotide rDNA sequence is repeated on both sides of the insertion. We discuss the variety of sequence rearrangements of the rDNA which flank type I insertions. PMID- 6273795 TI - Lac repressor - lac operator interaction. Circular dichroism study. AB - The interaction between lac repressor and a small operator DNA fragment have been examined by circular dichroism spectroscopy. The binding of lac repressor on the operator induces a conformation change of the DNA which is different from that observed upon non specific binding on non operator DNA. The CD titration curve indicates that the stoechiometry of interaction is complex. A two operators-one repressor complex was found. This result was confirmed by a gel filtration experiment. PMID- 6273797 TI - The distribution of benzo(a)pyrene DNA adducts in mammalian chromatin. AB - This paper describes the distribution of DNA-lesions generated by the potent carcinogen benzo(a)pyrene (BP) or its ultimate metabolic derivative 7 alpha, 8 8 beta, di-hydroxy-9 beta, 10 beta-epoxy-7,8,9,10-tetrahydrobenzo(a)pyrene (BPDE) within mammalian chromatin using the enzymic probe micrococcal nuclease. We have shown that the progress of the nuclease on naked DNA is unaffected by the presence of the hydrocarbon lesion at moderate extents of digestion. Digestion of nuclei isolated from murine erythroleukaemic cells immediately following BPDE treatment, and analysis of micrococcal nuclease resistant DNA by TCA precipitation, hydroxyapatite chromatography and gel electrophoresis demonstrates a non-random distribution of lesions. Approximately three times more binding occurs on the linker DNA regions between nucleosome cores than on the nucleosome core DNA itself. A similar result was obtained with BPDE treated primary mouse embryo cells; however nuclei isolated from these cells after prolonged treatment with BP (to allow metabolic activation) showed no such preferential binding. Post treatment incubation of BPDE-treated cells shows that this difference can be accounted for by the loss of preferential localisation with time. PMID- 6273798 TI - Interspersed repeated sequences in the African green monkey genome that are homologous to the human Alu family. AB - The dominant family of interspersed repetitive DNA sequences in the human genome has been termed the Alu family. We have found that more than 75% of the lambda phage in a recombinant library representing an African green monkey genome hybridize with a human Alu sequence under stringent conditions. A group of clones selected from the monkey library with probes other than the Alu sequence were analyzed for the presence and distribution of Alu family sequences. The analyses confirm the abundance of Alu sequences and demonstrate that more than one repeat unit is present in some phages. In the clones studied, the Alu units are separated by an average of 8 kilobase pairs of unrelated sequences. The nucleotide sequence of one monkey Alu sequence is reported and shown to resemble the human Alu sequences closely. Hence, the sequence, dispersion pattern, and copy number of the Alu family members are very similar in the African green monkey and human genomes. Among the clones investigated were two that contain segments of the satellite DNA term alpha-component joined to non alpha-component DNA. The experiments indicate that in the monkey genome Alu sequences can occur close to regions of alpha-component DNA. PMID- 6273799 TI - Adenovirus late sequences linked to herpes simplex virus thymidine kinase may be introduced into eukaryotic cells and transcribed. AB - LTK-cells have been transformed to the TK+ phenotype by treatment with size defined concatamers of HSV-1 TK DNA and Ad2 Bam H1 C fragment (42.0 - 59.5 map units). All TK+ transformants contained Ad2 DNA as well as HSV-1 TK sequences. In most cases several inserts of virus DNA were present, many in high copy numbers. Although no Ad2 transcription promoter was present in the transforming DNA, Ad2 specific sequences were detected in polyadenylated cytoplasmic RNA species from several cell lines. PMID- 6273800 TI - The nucleotide sequences of two tRNAAsn genes from tobacco chloroplasts. AB - Recombinant plasmids which contain EcoRI fragments of tobacco chloroplast DNA carrying tRNA genes were constructed. Plasmids pTC211 and pTC293 contain the base sequences for tRNAAsn in their 1.4 and 1.1 Md EcoRI fragments, respectively. These two tRNA sequences are identical and are; 5' TCCTCAGTAGCTCAGTGGTAGAGCGGTCGGCTGTTAACCGATTGGTCGTAGGTTCGAATCCTACTTGGGGAG-3'. Each tRNAAsn gene is located at about 0.9 kb apart from the distal end of each 5S rRNA gene and is coded for by the DNA strand opposite from that of the rRNA genes. PMID- 6273802 TI - The nucleotide sequence of the promoter and the amino-terminal region of alkaline phosphatase structural gene (phoA) of Escherichia coli. AB - The promoter and the amino-terminal region of phoA, the structural gene for alkaline phosphatase of Escherichia coli K12, was cloned by using a promoter cloning vector pMC1403. The nucleotide sequence of the cloned fragment has been determined. A sequence encoding the amino-terminal portion of mature alkaline phosphatase is found and it is preceded by a sequence encoding the signal peptide. The signal peptide consists of 21 amino acids; Met-Lys-Gln-Ser-Thr-Ile Ala-Leu-Ala-Leu-Leu-Pro-Leu-Leu-Phe-Thr-Pro-Val-Thr-Lys-Ala. The translation initiation codon is GUG, which is preceded by the Shine-Dalgarno sequence GGAG. Upstream to these sequences, there is a typical procaryotic promoter. TATAGTC for the Pribnow box. Around the Pribnow box, there are several dyad symmetrical sequences, which may probably be concerned with the regulation of this gene. PMID- 6273803 TI - Specific deletion of DNA sequences between preselected bases. AB - Blunt-end ligation of a "filled-in" HindIII, Sal I, Ava I or Bcl I restriction site with a DNA fragment having A, G, C, or T as the terminal 3' nucleotide regenerates the corresponding restriction site. A combination of this property with the action of BAL 31 nuclease which progressively removes base-pairs from the ends of linear DNA, can generate deletions extending to desired pre-selected nucleotides, and introduces unique restriction sites at those positions. Similarly other restriction sites can be used to select for the deletion of sequences between specific di-, tri-, tetra- and penta-nucleotides. Using this method, 10 base pairs were deleted from the end of a restriction fragment carrying the late promoter for bacteriophage T7 gene 1.1, to create a molecule with a unique restriction site at the initiation codon for translation. PMID- 6273801 TI - The structure and function of the regulatory elements of the Escherichia coli uvrB gene. AB - The construction and properties of recombinant plasmids carrying the Escherichia coli uvrB gene, including its transcriptional- and translational regulatory elements, is reported. The DNA sequence of the region, which governs the expression of the uvrB gene, has been determined. Within this sequence two non overlapping DNA segments match the model sequence for Escherichia coli promoters (1). The '-10 regions' and the '-35 regions' of the proposed uvrB promoters are, respectively, 5'TAAAAT (P1), 5'TATAAT (P2) and 5'TTGGCA (P1), 5'GTGATG (P2). The existence and the position of these promoters has been established by elimination of one promoter (P2), using molecular cloning procedures, by length measurements of in vitro synthesized 'run-off' transcripts and by protection of the uvrB regulatory region for S1 nuclease digestion using in vivo made RNA. Potential sites of interaction within the uvrB regulatory region with regulatory proteins, such as the LexA protein (2) and the UvrC protein (3) are discussed. PMID- 6273804 TI - Nucleotide sequence of the Aspergillus nidulans mitochondrial gene coding for the small ribosomal subunit RNA: homology to E. coli 16S rRNA. AB - The complete primary structure of the 1437 bp gene coding for mitochondrial 15S rRNA and its flanking regions was determined by Maxam-Gilbert sequencing of cloned HindIII fragment H3 of A. nidulans mtDNA. The gene product reveals significant homology (59%) to E. coli 16S rRNA, and the potential secondary structures of both rRNA molecules are very similar, except that the hairpin structures 7, 8 and 30 of the Brimacombe 16S rRNA model are deleted, and that two sequences of 8 and 31 nucleotides are inserted in the mitochondrial species. PMID- 6273805 TI - The high affinity binding site on polyoma virus DNA for the viral large-T protein. AB - In order to map the high affinity binding site for the viral large-T protein on polyoma virus DNA, we have developed an assay which does not require purified protein. It is based on the specific elution of the large-T ATPase activity from calf thymus DNA cellulose by recombinant DNA molecules including known sequences of the viral DNA. Using this assay, a high affinity binding site has been mapped on the early region side of the ori region. Binding requires the integrity of a sequence /AGAGGC/TTCC/AGAGGC/ (nucleotides 49 to 64 in the DNA sequence of the A2 strain). Similar repeats of a PuGPuGGC sequence within less than 20 bases are not found within the viral coding regions, but are strikingly common in the control regions of papovaviruses and other eukaryotic DNAs. PMID- 6273806 TI - Nucleotide sequence of the R.meliloti nitrogenase reductase (nifH) gene. AB - The nucleotide sequence of the structural gene (nifH) of nitrogenase reductase (Fe protein) from R.meliloti 41 with its flanking ends is reported. The amino acid sequence of nitrogenase reductase was deduced from the DNA sequence. The predicted R.meliloti nitrogenase reductase protein consists of 297 amino acid residues, has a molecular weight of 32,740 daltons and contains 5 cysteine residues. The codon usage in the nifH gene is presented. In the 5' flanking region, sequences resembling to consensus sequences of bacterial control regions were found. Comparison of the R.meliloti nifH nucleotide and amino acid sequences with those from different nitrogen-fixing organisms showed that the amino acid sequences are more conserved than the nucleotide sequences. This structural conservation of nitrogenase reductase may be related to its function and may explain the conservation of the nifH gene during evolution. PMID- 6273807 TI - DNA-like duplexes with repetitions. III. Efficient template-guided chemical polymerization of d(TGGCCAAGCTp). AB - Self-association of a decanucleotide d(TGGCCAAGCTp) in an aqueous solution is shown by UV spectroscopy, CD and sedimentation analysis to yield a pseudopolymeric (concatemeric) duplex having a geometry similar to that of DNA B type. It is demonstrated that in conditions when the concatemeric duplex is stable a water-soluble carbodiimide induces efficient polymerization of the 3'- or 5'-phosphorylated decanucleotide, and the resulting polymers d(TGGCCAAGCTp)2 10 contain only natural phosphodiester bonds. In conditions optimal for template guided polymerization of d(TGGCCAAGCTp) the overall yield of 20-100-member polynucleotides exceeds 90%. The obtained polymeric duplexes are cleaved by restriction endonuclease Alu II, Bsu RI, and Hind III to corresponding decamers which were isolated and sequenced. PMID- 6273809 TI - Nuclease sensitivity of chromatin containing active genes: kinetic analyses utilizing continuous elution of digestion products from an ultrafiltration cell. AB - Methods have been developed to analyze the kinetics of digestion of chromatin by nucleases. Radioactively labeled nuclei were incubated with enzyme in an ultrafiltration apparatus and digestion rates of different chromatin samples were computed employing a least-squares curve fitting technique to fit the data to zero-order and/or first-order kinetic models. These methods allow detailed kinetic analyses on small amounts of chromatin. Two biological systems were studied. 1) Tetrahymena thermophila macronuclei and micronuclei were compared; these nuclei differ in their transcriptional activities. 2) Ribosomal DNA (rDNA) of Tetrahymena pyriformis, approximately 60% of which codes for rRNA, can be preferentially labeled during starvation-refeeding; its digestion kinetics relative to bulk chromatin were studied. DNase I digested 20-40% of the macromolecular DNA about 3 times faster than bulk macronuclear or micronuclear DNA, and 60-80% of ribosomal gene-containing chromatin about 5 times faster than bulk chromatin. Filter hybridization studies of the DNAase I sensitivity of tRNA, 5S RNA, and ribosomal genes yielded similar results. These data are consistent with the observation that transcribed genes are especially sensitive to attach by DNase I and suggest that activated chromatin structure as probed by extensive DNase I digestion is the same in higher and lower eucaryotes for genes transcribed by all three RNA polymerases. Digestion kinetics of micrococcal nuclease were found to depend on the digestion conditions employed. These two biological systems and the methods we have developed should facilitate analyses of the factors responsible for maintaining an active chromatin structure. PMID- 6273808 TI - Different nucleotide changes in the large rRNA gene of the mitochondrial DNA confer chloramphenicol resistance on two human cell lines. AB - The nucleotide sequence of the mitochondrial DNA (mtDNA) in the region coding for the 3' end of the large rRNA has been determined for two human cell lines bearing independent cytoplasmic chloramphenicol-resistant (CAP-r) mutations. Comparison of the sequences of these two phenotypically different CAP-r mutants with their CAP-sensitive (CAP-s) parental cell lines has revealed a single base change for each in a region which is highly conserved among species. One CAP-r mutation is associated with an A to G transition on the coding strand while the second contains a G to T transversion 52 nucleotides away. Comparable sequence changes in this region had previously been found for mouse and yeast cell mitochondrial CAP-r mutants. Thus, changes in the large rRNA gene eliminate the inhibition of the ribosome by CAP and different nucleotide changes may result in variations in the drug-r phenotype. PMID- 6273810 TI - The effect of sequence specific DNA methylation on restriction endonuclease cleavage. AB - Sequence specific DNA methylation sometimes results in the protection of some or all of a restriction endonucleases' cleavage sites. This is usually, but not always, the result of methylation of one or both strands of DNA at the site characteristic of the corresponding "cognate" modification methylase. The known effects of sequence specific methylation on restriction endonucleases are compiled. PMID- 6273811 TI - The initiator tRNA genes of Drosophila melanogaster: evidence for a tRNA pseudogene. AB - We have isolated four segments of Drosophila melanogaster DNA that hybridize to homologous initiator tRNAMet. Three of the cloned fragments contain initiator tRNA genes, each of which can be transcribed in vitro. The fourth clone, pPW568, contains an initiator tRNA pseudogene which is not transcribed in vitro by RNA polymerase III. The pseudogene is contained in a 1.15 kb DNA fragment. This fragment has the characteristics of dispersed repetitive DNA and hybridizes in situ to at least 30 sites in the Drosophila genome. The arrangement of the initiator tRNA genes we have isolated, is different to that of other Drosophila tRNA gene families. The initiator tRNA genes are not clustered nor intermingled with other tRNA genes. They occur as single copies within an approximately 415-bp repeat segment, which is separated from other initiator tRNA genes by a mean distance of 17 kb. In situ hybridization to polytene chromosomes localizes these genes to the 61D region of the Drosophila genome. Hybridization analysis of genomic DNA indicates the presence of 8-9 non-allelic initiator tRNA genes in Drosophila melanogaster. PMID- 6273812 TI - Localization of putative transcription initiation site on the cloned rDNA fragment of Tetrahymena pyriformis. AB - A DNA fragment (1.4 Kb) which codes for 5' region of 35S ribosomal precursor RNA (pre-rRNA) in Tetrahymena pyriformis was cloned with pBR322. The fragment was cleaved from the central part of the palindromic rDNA with restriction endonuclease KpnI and HindIII, and ligated to the larger moiety of pBR322 DNA HindIII-BamHI fragment together with lambda DNA-KpnI-BamHI fragment through trimolecular ligation. The analysis of R-loop formed between KpnI-linearized recombinant plasmid and 35S pre-rRNA revealed a DNA:RNA hybrid region of 465 +/- 30 base pairs in length. Considering the contraction of DNA:RNA hybrids relative to DNA duplexes (Philippsen et al., J. Mol. Biol., 123, 387-404, 1978), the size of the hybrid region was corrected to about 490 base pairs. Alternatively, the size of DNA which was protected against nuclease S1 due to hybrid formation with 35S pre-rRNA was estimated to be 490 nucleotides long. These data indicate that the transcription initiation site is localized at about 490 base pairs from the HindIII site of the cloned rDNA fragment. PMID- 6273813 TI - A member of a new repeated sequence family which is conserved throughout eucaryotic evolution is found between the human delta and beta globin genes. AB - A new class of human interspersed repeated sequences distinct from the AluI family was found by screening a human gene library with a mouse ribosomal gene non-transcribed spacer probe (rDNA NTS). A member of this sequence family was localized to a 251 bp segment between the human delta and beta globin genes: a region previously judged to be devoid of repeated DNA. The complete nucleotide sequence of this segment revealed a tandem block of 17 TG dinucleotides, a feature hypothesized by others to be a recombination hot spot responsible for gene conversion in the gamma globin locus region. When the genomes of Xenopus, pigeon, slime mold and yeast were examined, reiterated sequences homologous to both the mouse rDNA NTS and human globin repeat were found in every case. The discovery of this extraordinarily conserved repeated sequence family appears to have depended upon not using salmon sperm DNA during hybridization. The use of eucaryotic carrier DNA may bias the search for repeated sequences against any which may be highly conserved during eucaryotic evolution. PMID- 6273814 TI - The appearance of DNase I hypersensitive sites at the 5' end of the late SV40 genes is correlated with the transcriptional switch. AB - DNase I digestion of the SV40 nuclear chromosome late in infection reveals three hypersensitive sites on the late side of the Bg1 I site. Two of these sites at bp 370, 270 correspond to the 5' side of the late transcripts while the third at bp 190, to a region that is required for early transcription. Early in infection, as well as in an SV40 transformed cell line and a T-Ag negative revertant (deleted in the coding region for T-Ag) only one of these sites is present - the one associated with early transcription. Thus, the positions of these major hypersensitive sites are related to the differential expression of the early and late genes. The presence of the characteristic hypersensitive site corresponding to "early" region expression in the revertant, where large T antigen is not synthesized, but where the early "promotor" is intact, indicates that large T antigen is not responsible for this particular hypersensitive site. Additional minor specific DNase I cuts were found on the early genes, at early times only, at 300, 550, 850 bp from Bg1 I site. In the transformed cell line, one of these minor cuts is found about 350-400 bp from the Bg1 I site and in the revertant, where this region is deleted, a new site is created at 100 bp. PMID- 6273815 TI - Isolation and characterization of cloned rat DNA fragment carrying tRNA genes. AB - A rat genomic library was screened for tRNA genes with an unfractionated rat liver tRNA probe. About 70 clones containing tRNA genes were detected per rat genome. The organization of tRNA genes in five clones was analyzed by restriction endonuclease digestion, RNA-DNA hybridization and in vitro transcription with nuclear extracts from Xenopus oocytes. Evidence is presented suggesting that tRNA genes are distributed in the rat genome in small clusters spanning 1 to 2 kb and interspersed with large regions (minimum 8 to 20 kb) of non tRNA-coding DNA. The tRNA gene clusters were found to contain the sequences for a variety of tRNA species. Genes for a single isoacceptor, were found in more than one clone. The detailed study of one clone shows the repetition of a cluster of four tRNA sequences at a distance of about 8 kb. The arrangement of tRNA genes in rat appears to follow the irregular pattern of tRNA gene organization previously reported in Drosophila and Xenopus. PMID- 6273816 TI - Nucleotide sequence of a Bacillus subtilis promoter recognized by Bacillus subtilis RNA polymerase containing sigma 37. AB - We report the nucleotide sequence of the promoter for a Bacillus subtilis gene (spoVC) whose transcription is controlled by a 37,000 dalton species of B. subtilis sigma factor known as sigma 37 but not by the principal- sigma factor of 55,000 daltons (sigma 55). Using S1 nuclease mapping we show that the startpoint for sigma 37-directed transcription of the spoVC gene in vitro corresponded closely to the 5' terminus of in vivo synthesized spoVC RNA. The binding site for sigma 37-containing RNA polymerase extended from 43 bp to 51 bp (positions -43 to -51) upstream from the transcription startpoint to 22 bp (position +22) downstream from the startpoint. The nucleotide sequence of the spoVC promoter differed significantly from promoters whose recognition is controlled by sigma 55 but was similar to other sigma 37- controlled promoters in regions known to be important in promoter recognition. Our results are consistent with the hypothesis (Lee and Pero, J. Mol. Biol., in press) that sigma factors work by contacting specific bases in both the -35 and -10 regions of promoters. PMID- 6273817 TI - Nucleotide sequences of two Bacillus subtilis promoters used by Bacillus subtilis sigma-28 RNA polymerase. AB - RNA polymerase holoenzymes from many bacterial species share a common promoter recognition specificity since they use the same promoter sites on a variety of templates. These promoters generally include sequences homologous to the average sequences -TTGACA- and -TATAATA-, located -35 and -10 base pairs, respectively, upstream of the transcriptional state site. We have isolated a minor form of B. subtilis RNA polymerase in which the normal sigma subunit (sigma 55) is replaced by a smaller polypeptide (sigma 28) and which is highly specific for a class of promoter sites not used by the sigma 55-holoenzyme. Sequencing of two B. subtilis promoter sites used by the sigma 28-holoenzyme reveals identical sequences at -35 and -10 base pairs from the start site, which are -CTAAA- and -CCGATAT-, respectively. These results confirm that sigma subunit plays a major direct role in promoter sequence recognition, and support a model in which sigma interacts sequentially with -35 and -10 regions, respectively. PMID- 6273818 TI - In situ photochemical crosslinking of HeLa cell mitochondrial DNA by a psoralen derivative reveals a protected region near the origin of replication. AB - The in vivo association with proteins of HeLa cell mitochondrial DNA (mtDNA) has been investigated by analyzing the pattern of in situ crosslinking of the DNA by 4'-hydroxymethyl-4, 5',8-trimethylpsoralen (HMT). Either isolated mitochondria or whole cells were irradiated with long wavelength UV light in the presence of ths psoralen derivative, and the mtDNA was then isolated and analyzed in the electron microscope under totally denaturing conditions. No evidence of nucleosomal structure was found. The great majority of the molecules (approximately 90%) had a double-stranded DNA appearance over most of their contour length, with one to several bubbles occupying the rest of the contour, while the remaining 10% of the molecules appeared to be double-stranded over their entire length. Analysis of restriction fragments indicated the presence, in approximately 80% of the molecules, of a protected segment (300 to 1500 bp long) in a region which was centered asymmetrically around the origin of replication so as to overlap extensively the D-loop. Control experiments showed that at most 30% of the bubbles found near the origin could represent D-loops or expanded D-loops: furthermore, it could be excluded that some sequence peculiarity would account for the preferential location of bubbles near the origin of replication. The data have been interpreted to indicate that, in at least 55% of HeLa cell mtDNA molecules, the region around the origin is protected from in situ psoralen crosslinking by proteins or protein complexes which are associated in vivo with the DNA. PMID- 6273819 TI - The interaction of RNA polymerase II with non-promoter DNA sites. AB - Various complexes formed between purified RNA polymerase II and simian virus 40 DNA have been characterized with respect to rates of formation, rates of dissociation, and initial velocity of RNA synthesis. Two different types of complexes can form on intact DNA templates. One of these is formed rapidly, but is quite labile; the other forms more slowly, but is moderately stable once formed. The introduction of a single strand break into DNA leads to rapid and stable complex formation, and thus is expected to create the favored binding site. The observed properties of these complexes provide a general framework for describing the interactions of RNA polymerase II at non-promoter DNA sites. This framework appears to be similar to that established for Escherichia coli RNA polymerase interactions, suggesting that the fundamental mode of non-promoter DNA binding is similar for the bacterial, plant, and mammalian enzymes. PMID- 6273820 TI - The SV40 72 base repair repeat has a striking effect on gene expression both in SV40 and other chimeric recombinants. AB - By introduction of recombinant plasmids into monkey CV1 cells, we have unambiguously demonstrated that sequences entirely within the 72 bp repeat, which is located upstream of the SV40 early region, are crucial for T-antigen expression in vivo. We have also shown that a DNA fragment containing the 72 bp repeat, inserted directly before chicken conalbumin or adenovirus-2 major late promoter sequences in chimeric plasmids where these promoters replace that of the SV40 early genes, caused a dramatic increase in the expression of T-antigen in vivo. This effect was independent of the orientation of the 72 bp repeat, but was sensitive to its location within the plasmid, when the 72 bp repeat was separated from the promoter sequences, T-antigen expression was reduced. Insertion of the 72 bp repeat into equivalent plasmids containing no known eukaryotic promoter sequences (plasmids which were not detectably expressed in vivo) gave rise to a measurable, but smaller level of expression. The stimulation of expression by the 72 bp repeat is cis-acting : it required covalent linkage to the recombinant. We discuss the possibility that the 72 bp repeat region in SV40 may act as a bi directional entry site for RNA polymerase B such that promoter sequences linked to the repeat are more efficiently utilised. PMID- 6273821 TI - The primary structure of bovine satellite 1.715. AB - The primary structure of the 1402 bp repeat unit of bovine satellite 1.715 has been determined using a dimer inserted at the SalI site of plasmid pBR322 and cloned in E. coli. In contrast with bovine satellites 1.706, 1.720b and 1.711a, the 1.715 satellite has a complex sequence with no obvious internal short prototype repeat. The sequence consists however of repeats ranging in length from 6 to 13 nucleotides. In addition, the hexanucleotide, AGATGA, present in the prototype sequences of satellites 1.706, 1.720b and 1.711a, is found in satellite 1.715 in repeats as long as, or longer than, 8 nucleotides, establishing a homology link among those satellites on one hand and satellite 1.715 (and the related satellite 1.711b) on the other. In turn, this suggests a common evolutionary origin. A comparison of the maps for 15 restriction enzymes of cloned and uncloned satellite indicates very little sequence divergence among the repeat units of the latter, most of the differences being due to methylation. PMID- 6273823 TI - Mapping of a mouse ribosomal DNA promoter by in vitro transcription. AB - An in vitro transcription system that provides proper initiation of RNA polymerase I on cloned rDNA has been used to identify the start site for rDNA transcription. Different subclones that span defined regions of the 5' terminal region of the ribosomal gene have been constructed and assayed in the cell-free system for their ability to promote specific initiation of pre-rRNA synthesis. It is shown that rapid processing at the 5' end of the primary transcript occurs both in vivo and in vitro which in former studies has led to a wrong interpretation of the S1 nuclease mapping data (1 - 3). RNA polymerase I starts in vitro at a unique point on the rDNA yielding run-off transcripts that have a triphosphorylated 5' end pppApC. If multiple copies of the promoter-containing rDNA fragment were placed in head-to-tail orientation in front of the transcribed region distinct RNA products were synthesized that have been started at the tandem initiation sites. Removal of sequences upstream the initiation site indicates that 5' flanking regions are essential for specific transcription. PMID- 6273822 TI - Selective inhibition by harmane of the apurinic apyrimidinic endonuclease activity of phage T4-induced UV endonuclease. AB - 1-Methyl-9H-pyrido-[3,4-b]indole (harmane) inhibits the apurinic/apyrimidinic (AP) endonuclease activity of the UV endonuclease induced by phage T4, whereas it stimulates the pyrimidine dimer-DNA glycosylase activity of that enzyme. E. coli endonuclease IV, E. coli endonuclease VI (the AP endonuclease activity associated with E. coli exonuclease III), and E. coli uracil-DNA glycosylase were not inhibited by harmane. Human fibroblast AP endonucleases I and II also were only slightly inhibited. Therefore, harmane is neither a general inhibitor of AP endonucleases, nor a general inhibitor of Class I AP endonucleases which incise DNA on the 3'-side of AP sites. However, E. coli endonuclease III and its associated dihydroxythymine-DNA glycosylase activity were both inhibited by harmane. This observation suggests that harmane may inhibit only AP endonucleases which have associated glycosylase activities. PMID- 6273824 TI - The effect of several nucleic acid binding drugs on the cleavage of d(GGAATTCC) and pBR 322 by the Eco RI restriction endonuclease. AB - The endonucleolytic action of the EcoRI restriction enzyme on the double-stranded oligonucleotide d(GGAATTCC) and the supercoiled plasmid DNA pBR 233 is inhibited by actinomycin D, ethidium bromide, proflavin, distamycin A and netropsin. Half maximal inhibition is observed at around 100 microM concentrations for the intercalating drugs, and around 0.1 to 1 microM concentrations for netropsin and distamycin A. The inhibitory activity of these drugs can be correlated with their affinity to the oligonucleotide and the plasmid DNA. Since at high concentrations of the drugs a complete inhibition is observed, it is concluded that the effect of the drugs on the stereochemistry of the EcoRI site is such that recognition is excluded. PMID- 6273825 TI - An unusual 5S rRNA, from Sulfolobus acidocaldarius, and its implications for a general 5S rRNA structure. AB - The nucleotide sequence of the 5S ribosomal RNA of the thermoacidophilic archaebacterium Sulfolobus acidocaldarius was determined. The high degree of evident secondary structure in the molecule has implications for the common higher order structure of other 5S rRNAs, both bacterial and eukaryotic. PMID- 6273826 TI - Recovery of recombinant bacterial plasmids from E. coli transformed with DNA from microinjected mouse cells. AB - We have previously described the isolation of thymidine kinase positive (TK+), human beta-globin gene-containing colonies following co-microinjection of mouse TK- L cells with two recombinant pBR322 plasmids, one containing the TK gene of herpes simplex virus type I (plasmid pXl), and the second containing a human genomic DNA fragment within which is the human beta-globin gene (plasmid pRKl). DNA isolated from one such clone was used in bacterial transformation experiments with a selection for tetracycline-resistant colonies (that is, for cells containing pRKl). A total of forty-two tetracycline-resistant colonies were isolated, thirty of which contained circular pRK1 molecules identical to those originally injected. The remaining twelve colonies contained unique plasmids that were grouped into five different classes of recombinant molecules. All five of these unique recombinant classes appear to contain a common deletion endpoint occurring at a specific region of the pBR322 segment of pRKl. Four of the unique recombinant classes appear to have arisen from the deletion of a segment of a pRKl trimer or dimer molecule, while the fifth class appears to have resulted from recombination between pRKl and pXl followed by a deletion event within this recombinant. It is uncertain whether these deletions are occurring within the eukaryotic cell or upon subsequent transformation of the bacterial cell. If the latter, then the passage of the plasmid DNA through the eukaryotic cell alters a specific site of the pBR322 DNA in such a way that deletions can occur at a high frequency in this region when the plasmid DNA is introduced back into a bacterial cell. Thus, we have established a prokaryote-eukaryote-prokaryote DNA transfer and recovery system which should be useful in studies on DNA replication and the regulation of gene expression in higher eukaryotes. PMID- 6273827 TI - [Methodological problems of sacroiliac joint scintigraphy (author's transl)]. AB - Bone scintigraphy of the sacroiliac joints and the lumbar spine was performed in 35 patients with ankylosing spondylitis and 30 control subjects. The scans were evaluated by qualitative and quantitative assessment. The sacroiliac/sacrum ratio (index ISG/sacrum), the sacroiliac/lumbar spine ratio (index ISG/LWS), and the left sacroiliac joint/right sacroiliac joint ratio (index li. ISG/re. ISG) were calculated. The visual interpretation of colour scans proved to be unreliable. The index ISG/sacrum was more sensitive than the index ISG/LWS. The index li. ISG/re ISG was helpful as an additional criterion. When digital sacroiliac joint scintigraphy is well standardized it can be considered as a useful technique in early diagnosis of ankylosing spondylitis. PMID- 6273828 TI - Nucleotide requirements for DNA replication of SV40 chromatin in digitonin treated and saponin-treated permeable cells. AB - Permeable cell systems have been developed by treatment with digitonin or saponin for studying in vitro DNA replication of chromatin. DNA replication of SV40 nucleoprotein complexes (SV40 chromatin) in these systems requires the addition of ATP, dCTP, dGTP, dTTP, and Mg2+. Further addition of three other ribonucleoside triphosphates slightly increased DNA synthesis. Omission of ATP greatly reduced DNA synthesis. In the presence of ATP, however, omission of dATP did not significantly alter or rather slightly enhanced DNA synthesis. Analysis of replicated SV40 DNA by gel electrophoresis and autoradiography indicated that complete replication of SV40 DNA and chromatin occurred in these systems PMID- 6273829 TI - Methylation of 5'-5' confronting dinucleotides by vaccinia associated enzyme system. AB - Methylation of a series of the 5'-5' confronting dinucleotides containing various number of phosphates was examined by the use of virion enzyme system of vaccinia virus. Methylation was absolutely dependent on the number of interposed phosphate groups between 5'-5' confronting dinucleotides, showing maximum efficiency in the 3 phosphate compound corresponding to the blocked 5'-terminus (cap) of an eukaryotic mRNA. Methylation, if any, occurred only at the 7-position of guanine residue, but not at the 2' position of ribose moiety. PMID- 6273830 TI - Search for new antiamebic agents. I. Synthesis and bioassay of new Schiff bases of 1-phenyl-2-morpholinoethylidene-1-urea. AB - As potential agents against E. histolytica 19 new Schiff bases of 1-phenyl-2 morpholinoethylidene-1-urea were synthesized by the condensation of the parent compounds with various carbonyl compounds. Thirteen of them were tested against the axenic culture of E. histolytica at concentration 125 microgram/cm3 but no significant amoebicidal activity was observed. PMID- 6273831 TI - Diabetic ketoalkalosis due to ectopic ACTH production from an oat cell carcinoma. AB - A diabetic patient is described who presented with the bizarre biochemical association of hyperglycaemia, ketosis and severe alkalosis. Investigation revealed hypercortisolaemia due to ectopic adrenocorticotrophic hormone secretion from an oat cell carcinoma of the bronchus and post-mortem studies demonstrated clinically unsuspected resolving pancreatitis. a possible mechanism for the metabolic upset is discussed. PMID- 6273832 TI - Malignant islet-cell tumour of the pancreas presenting with non-bacterial thrombotic endocarditis and eosinophilia. AB - A case of severe persistent eosinophilia and non-bacterial thrombotic endocarditis complicating a malignant islet-cell tumour of the pancreas is documented. Although these are well recognized complications of malignant tumours, they do not appear to have been previously documented from the same patient. These complications probably reflect ectopic synthesis and are unlikely to be causally interrelated. PMID- 6273833 TI - Response of ectopic prostatic ACTH production to metyrapone. PMID- 6273834 TI - [Erythrocytic enzymopathies associated with hemolytic anemia]. PMID- 6273835 TI - [Use water-soluble and lipid-bound forms of cytochrome C for the treatment of experimental hemorrhagic shock]. PMID- 6273836 TI - Molecular cloning of the genome of poliovirus type 1. AB - Poliovirus cDNA.RNA hybrids were prepared from the Mahoney strain of poliovirus type 1 by using reverse transcriptase (RNA-dependent DNA nucleotidyltransferase) and cloned in the Escherichia coli plasmid pBR322. Bacteria colonies carrying recombinant plasmids were selected by in situ hybridization with virus-specific RNase T1-resistant oligonucleotides. Analysis of the cDNA inserts by restriction mapping and electron microscopy showed that the cloned cDNAs, the longest of which was 3.2 kilobase pairs, originated from various parts of the viral RNA, covering at least 99% of the genome length. Due to overlapping of the clones, the restriction map of the poliovirus genome could be reconstructed. The complete 5' end of the genome was successfully cloned in at least one of the recombinant plasmids, pPV1-366. PMID- 6273837 TI - Adult chicken alpha-globin genes alpha A and alpha D: no anemic shock alpha globin exists in domestic chickens. AB - Three alpha-type globin genes have been identified in the alpha-globin linkage group of chickens. No other alpha-type genes hae been directly shown to be within 10 kilobase pairs of any of these three closely linked genes. These three genes have been conclusively identified by DNA sequence analysis. The gene at the 5' end of the linkage group is an embryonic alpha-type globin gene, pi or pi', and the central gene corresponds to the minor adult alpha- globin, alpha D. The 3' terminal gene sequence corresponds to the sequence of cDNA clones previously described as "alpha S", presumed anemic shock-induced alpha-globin gene [Salser, W. A., Cummings, I., Liu, A., Strommer, J., Padayatty, J. & Clarke, P. (1979) in Cellular and Molecular Regulation of Hemoglobin Switching, eds. Stamatoyannopoulos, G. & Nienheis, A. (Grune and Stratton, New York), pp. 621 643; Richards, R. I. & Wells, J. R. E. (1980) J. Biol. Chem. 255, 9306-9311]. Several groups of workers have isolated alpha S-type cDNA clones but no one has identified a cDNA clone corresponding to the published amino acid sequence of the major chicken alpha-globin, alpha A. We have identified the alpha S-type sequence as the only abundant alpha-like globin sequence in cDNA clones made from reticulocyte mRNA isolated from nonanemic chickens. Therefore, we suggest that the alpha S-type sequence corresponds to the true alpha A-globin species. PMID- 6273838 TI - Characterization of sites for tyrosine phosphorylation in the transforming protein of Rous sarcoma virus (pp60v-src) and its normal cellular homologue (pp60c-src). AB - The transforming protein of Rous sarcoma virus (pp60v-src) and its normal cellular homologue (pp60c-src) appear to be protein kinases that phosphorylate tyrosine in a variety of protein substrates. In addition, pp60v-src and pp60-c src are themselves phosphorylated on serine and tyrosine. It is likely that these phosphorylations serve to regulate the function(s) of pp60v-src and pp60c-src. We have therefore characterized the sites of tyrosine phosphorylation in the two proteins. Tyrosine phosphorylation of pp60v-src in infected cells occurs mainly (if not entirely) at residue 419 in the deduced amino acid sequence of the protein. Surrounding this residue is the sequence Leu-Ile-Glu-Asp-Asn-Glu-Tyr(P) Thr-Ala-Arg. This peptide is distinguished by the fact that three out of the four amino acids that precede the phosphorylated tyrosine are acidic in nature. These results define what may prove to be a widely used site for tyrosine phosphorylation in the regulation of cellular function. The same site was phosphorylated when partially purified pp60v-src was used in a phosphotransfer reaction in vitro. The results with pp60c-src were more complex. The site of tyrosine phosphorylation in vitro appeared to be the same as that found in pp60v src. By contrast, phosphorylation of pp60c-src in vivo apparently occurred at a different, and currently unidentified, tyrosine residue. It is therefore possible that pp60v-src and pp60c-src respond differently to regulatory influences in the intact cell. PMID- 6273839 TI - Directionality and polarity in recA protein-promoted branch migration. AB - The recA protein of Escherichia coli promotes the complete exchange of strands between full-length linear duplex and single-stranded circular phi X174 DNA molecules. Analysis of the reaction by electron microscopy confirms that D loops containing short heteroduplex regions are rapidly formed at the ends of the linear duplex, followed by a relatively slow branch migration that converts the D loops to nicked circular duplexes (RFII) and displaced linear single strands. Heteroduplex extension and displacement of the linear single strand are concerted. Heterologous sequences within the linear duplex halt branch migration and lead to the accumulation of D loops. Although D loops can be formed at either end of the linear duplex, recA protein-promoted branch migration proceeds uniquely in the 3' leads to 5' direction relative to the (--) strand of the linear duplex. PMID- 6273840 TI - Mechanisms of Kirsten murine sarcoma virus transformation-induced changes in the collagen phenotype and synthetic rate of BALB 3T3 cells. AB - Specific viral transformation rather than cell selection can explain the previously observed increase in the proportion of type III procollagen compared to type I procollagen in BALB 3T3 cells transformed by Kirsten murine sarcoma virus (Ki-MSV). Two subclones of BALB 3T3 A31 were productively infected with with a temperature-sensitive Ki-MSV in the presence of helper murine leukemia virus (MLV), resulting in virtually complete transformation of cultures and eliminating selection of transformed foci. Analysis of radioactive collagen, derived from procollagen by pepsin treatment, showed that both of the tsKi MSV/MLV-transformed subclones contained a 4-fold greater proportion of type III procollagen than did control MLV-infected cultures. A nonproducer derivative exhibited an even greater change (10-fold), indicating that viral replication was irrelevant. After 48 hr at a nonpermissive temperature, tsKi-MSV-transformed cells retained a high proportion of type III procollagen, suggesting that either this change is not induced by src protein or else there is a slowly reversible or irreversible step involved. Alternatively, type III procollagen mRNA may be long lived. In contrast, the relative rate of procollagen synthesis in transformed cells was clearly regulated by src protein. Translation of mRNA from cells preincubated at permissive or nonpermissive temperatures revealed that the decreased relative rate can be explained by a simultaneous small decrease in the level of procollagen mRNA and a large increase in mRNA for noncollagen proteins. PMID- 6273841 TI - Cloning of the uvrC gene of Escherichia coli: expression of a DNA repair gene. AB - We have cloned the uvrC gene of Escherichia coli, using an F' plasmid carrying the uvrC region as a source of DNA. Two plasmids, pSC101 and pBR322, were used as cloning vectors. The recombinant plasmids were selected for their ability to complement the uvrC defect of E. coli strains AB1884 and N177. We conclude that the uvrC structural gene is contained in a 1.9-kilobase DNA fragment. The protein encoded by the uvrC gene appears to have a monomer molecular weight of 64,500 as analyzed by denaturing polyacrylamide gel electrophoresis. Strains containing multicopy uvrC+ plasmids overproduce a factor that is missing in lysates of uvrC- mutants and required for an in vitro model repair reaction. The expression of uvrC+ hybrid plasmids suggests that the structural gene is separated by at least 0.8 kilobase from the regulatory region. PMID- 6273842 TI - Two periplasmic transport proteins which interact with a common membrane receptor show extensive homology: complete nucleotide sequences. AB - The hisJ and argT genes of Salmonella typhimurium encode two periplasmic binding proteins, J and LAO, which are involved in histidine and arginine transport, respectively, and which interact with a common membrane-bound component, the P protein. The complete nucleotide sequences of these two genes have been determined. The two genes show extensive homology (70%) and presumably arose by tandem duplication of a single ancestral gene. The two encoded proteins now perform distinct functions but still retain sufficient homology to permit interaction with the same site on the membrane-bound P protein. Three lines of evidence have allowed both the amino acid-binding site and the site involved in the interaction with the P protein to be assigned to specific regions of each binding protein: (i) the distribution of amino acid differences between the two proteins; (ii) the properties of a functional chimeric protein, produced by a deletion mutant in which the first half of the argT gene is fused to the second half of the hisJ gene; (iii) the sequence change in a mutant J protein unable to interact with P. PMID- 6273843 TI - Methotrexate-resistant Chinese hamster ovary cells have amplified a 135-kilobase pair region that includes the dihydrofolate reductase gene. AB - For the eventual purpose of isolating and studying a single animal cell replicon, we have developed a methotrexate-resistant Chinese hamster ovary cell line that has amplified an early-replicating DNA sequence approximately 500 times; this sequence includes the gene coding for dihydrofolate reductase (DHFR; tetrahydrofolate dehydrogenase; 5,6,7,8-tetrahydrofolate:NADP+ oxidoreductase, EC 1.5.1.3). DHFR composes 30% of the cytoplasmic protein in this cell line, and DHFR mRNA represents 25% of the message translatable in vitro. After digestion of genomic DNA from resistant cells with restriction enzymes, a unique set of highly repetitive restriction fragments can be visualized on agarose gels by ethidium bromide staining. These bands are not present in digests of parental DNA. We estimate the total length of the unit repeated sequence to be 135 +/- 15 kilobase pairs. Regardless of the restriction enzyme utilized, a subset of these repetitive fragments hybridizes to radioactive DHFR cDNA. The homogeneously staining regions on mitotic chromosomes in which these amplified sequences are located are shown to be early-replicating, as are the highly repeated restriction fragments themselves. These data suggest that an early replicon can be isolated from this region, and that this entire, normally unique, genomic segment can be cloned and mapped with respect to origins of DNA synthesis and promoters for transcription, as well as other genetic features of interest. PMID- 6273844 TI - Fluorometric assay for adenosine 3',5'-cyclic monophosphate-dependent protein kinase and phosphoprotein phosphatase activities. AB - A novel peptide substrate for adenosine 3',5'-cyclic monophosphate-dependent protein kinase (ATP:protein phosphotransferase, EC 2.7.1.37), Leu-Arg-Arg-Trp-Ser Leu-Gly, was synthesized. Phosphorylation of the peptide causes a 20% increase in the peptide fluorescence intensity at 358 nm. Values of Km and kcat for the phosphorylation reaction at pH 7.0 (25 degrees C), were determined to be 2.7 +/- 0.5 microM and 5.5 +/- 0.4 sec-1, respectively. The phosphorylated peptide was shown to be an effective substrate for phosphoprotein phosphatase (phosphoprotein phosphohydrolase, EC 3.1.3.16) with a Km of 113 +/- 10 microM and a kcat of 2.4 +/- 0.2 sec-1 in the presence of 2.5 mM MnCl2. Changes in the peptide fluorescence intensity as a function of its phosphorylation state provide a highly sensitive assay of cyclic AMP-dependent protein kinase and phosphoprotein phosphatase activities. PMID- 6273845 TI - Membrane glycoproteins involved in cell--substratum adhesion. AB - A combination of immunological and biochemical methods were used to identify surface membrane components involved in cell-substratum adhesion. Broad-spectrum antiserum, prepared against surface membranes from hamster cells, induced reversible rounding and detachment of hamster fibroblasts from a substratum in vitro. This phenomenon was inhibited by Nonidet P-40 extracts of hamster cells. Therefore, an antibody neutralization assay was developed to detect the presence of antigen during the fractionation of Nonidet P-40 extracts of cells. After two differential precipitation steps, anion exchange chromatography, and sequential lectin affinity chromatography, a fraction greatly enriched in ability to block antiserum-induced changes in cell adhesion and appearance was isolated. Analysis of this fraction by NaDodSO4/polyacrylamide gel electrophoresis revealed a highly restricted group of glycoproteins with Mr approximately 140,000. A lectin purified glycoprotein fraction was used to raise a higher titer antiserum that was able to induce reversible rounding and detachment of cells from a substratum and, when immobilized on an antibody affinity column, was able to bind and release material capable of blocking antiserum-induced cell rounding. These methods have allowed us to focus attention on a restricted group of glycoproteins that are integral constituents of the surface membrane and which play some as yet undetermined role in the process of cell--substratum adhesion. PMID- 6273846 TI - Use of whole-cell fixation to visualize replicating and maturing simian virus 40: identification of new viral gene product. AB - Formaldehyde fixation of simian virus 40 (SV40)-infected CV-1 cells at appropriate times after infection permits us to isolate crosslinked complexes of SV40 minichromosomes during the time of DNA replication and during packaging with viral proteins. Such crosslinked complexes can be separated on the basis of density on CsCl/guanidine . HCl density gradients. During the course of these studies we observed the presence of a low molecular weight protein in a region of the gradient much enriched with viral nucleoproteins. This protein is present only in infected cells and has a molecular weight and amino acid composition consistent with it being the product of the so-called SV40 agnogene. PMID- 6273848 TI - Isolation of the Escherichia coli leader peptidase gene and effects of leader peptidase overproduction in vivo. AB - The only covalent modifications known to accompany protein insertion into membranes or protein secretion are glycosylation and the proteolytic removal of an NH2-terminal leader (signal) sequence. This latter reaction is catalyzed by leader peptidase, a constitutive, membrane-bound proteinase. We now report the identification of a plasmid-bearing strain of Escherichia coli that overproduces leader peptidase 4- to 6-fold. This strain grows normally and shows an unaltered polypeptide composition of inner ad outer membranes. The leader peptidase gene has been subcloned and transferred from this plasmid to the multicopy plasmid pBR322, yielding a new plasmid (pTD101). Strains transformed by pTD101 have a 30 fold increase in leader peptidase. We have studied the effect of leader peptidase overproduction on the insertion of newlymade M13 phage coat protein into the plasma membrane of infected cells. The overproducer strain, when infected by M13 phage, shows a dramatic acceleration in the conversion of procoat (a cytoplasmic precursor form) to coat (an integral, transmembrane protein). Thus the leader peptidase that converts M13 procoat to coat in vitro can catalyze this reaction in vivo as well. PMID- 6273847 TI - The Mr 80,000 common forms of neurophysin and vasopressin from bovine neurohypophysis have corticotropin- and beta-endorphin-like sequences and liberate by proteolysis biologically active corticotropin. AB - We have tested the hypothesis that the high M(r) forms common to both neurophysin and vasopressin detected in bovine neurohypophysis extracts (Nicolas, P., Camier, M., Lauber, M., Masse, M.-J. O., Mohring, J. & Cohen, P. (1980) Proc. Natl. Acad. Sci. USA 77, 2587-2591) might also contain the sequences of other known neuropeptides. The following evidence indicates that corticotropin- and beta endorphin-like sequences are associated with similar high M(r) forms and are included in these M(r) 80,000 molecules. During the fractionation steps of high M(r) material, both corticotropin and beta-endorphin immunoreactive species were found to coelute with the neurophysin and vasopressin ones, either under M(r) 140,000 (in 0.1 M formic acid) or M(r) 70,000-80,000 (in 6 M guanidine) elution volumes. Corticotropin immunoreactivity was found to cofocus at pIs 6.05 and 5.8 with the M(r) 80,000 neurophysin-containing species. This material was submitted to affinity chromatography on purified anti-neurophysin antibodies covalently attached to Sepharose 4B. Both the corticotropin and beta-endorphin immunoreactivities, together with the neurophysin and vasopressin immunoreactivities, were retained on the immunoadsorbent and codesorbed by either a drastic pH change or by selective displacement with an excess of neurophysin. Comparison of the tryptic-digest maps of either the M(r) 68,000 fragment immunoprecipitated by anti-corticotropin antibodies or the M(r) 68,000 fragment released after precipitation of the M(r) 80,000 species by anti-neurophysin antibodies indicated large sequence homologies. Exposure of either the M(r) 80,000 or 68,000 components to mild proteolytic activities resulted in the formation of lower-size fragments. The resulting corticotropin-like immunoreactive material, recovered under the elution volume of standard (125)I labeled corticotropin-(1-24), was tested for its ability to activate glucocorticoid biogenesis by the amphibian interrenal tissue (adrenal) in perifusion. It was found to exhibit a noticeable activity qualitatively undistinguishable from the one of the reference human corticotropin-(1-39). The name neurohypophyseal "coenophorin" (from the Greek word for common) is proposed for this class of M(r) 80,000 polypeptides that might represent the common precursor store-house for a set of neuropeptides produced in the hypothalamo neurohypophyseal tract. PMID- 6273849 TI - Escherichia coli factor Y sites of plasmid pBR322 can function as origins of DNA replication. AB - The Escherichia coli replication factor Y, in conjunction with other genetically undefined E. coli replication factors and the gene products of the E. coli dnaB, dnaC, and dnaG loci, is involved in de novo primer formation on the phi X174 (+) single-strand circular DNA template [(+) ss(c)DNA]. The participation of factor Y in this series of reactions is correlated with its phi X174 (+) ss(c)-specific DNA-dependent ATPase activity. Recently two factor Y effector DNA segments of the plasmid pBR322 have been identified in close proximity to the plasmid origin of DNA replication. We report here that insertion of these factor Y sites into the filamentous phage f1R229 (+) ss(c)DNA confers upon it the ability to be converted to RF DNA in vitro through a rifampicin-resistant dnaB, dnaC, and dnaG gene product-dependent pathway. Our data suggest that factor Y effector sites can function as origins of DNA replication. PMID- 6273850 TI - Elongation of displacement-loop strands in human and mouse mitochondrial DNA is arrested near specific template sequences. AB - Animal mitochondrial DNA (mtDNA) maintains a displacement loop (D loop) at the heavy strand origin of replication. These D loops represent sharply limited synthesis of heavy strands and provide a unique opportunity to examine the termination of DNA synthesis. Direct sizing at the nucleotide level indicates that the 3' ends of D-loop strands of human and mouse mtDNA are discrete and map within three to five nucleotides on the complementary template strand. In the case of human mtDNA, there is a single trinucleotide stop point 51-53 nucleotides downstream from a 15-nucleotide template sequence (3'T-A-A-C-C-C-A-A-A-A-A-T-A-C A 5') which is repeated four times in the mouse mtDNA D-loop region 3'(T-A-A-Py Py-A-A-A-T-T-A-C-A 5'). The stop points of the five major mouse D-loop strands are 24-63 nucleotides downstream from the four repeated template sequences. These results suggest that the arrest of D-loop strand elongation is an event determined by template sequence. PMID- 6273851 TI - Human somatotropin: covalent reconstitution of two polypeptide contiguous fragments with thrombin. AB - Thrombin has been shown to resynthesize the Arg134-Thr135 peptide bond between the 134- and 57-residue thrombin fragments of reduced and carbamidomethylated human somatotropin at pH 6.0 in 90% (vol/vol) glycerol. The maximal amount of synthesis was about 20% as estimated by sodium dodecyl sulfate gel electrophoresis and radioreceptor assay. The resynthesized polypeptide was isolated and shown to be indistinguishable from the reduced and carbamidomethylated hormone when tested by two receptor-binding assays, radioimmunoassay, and rat tibia test. PMID- 6273852 TI - Uninfected cell polymerase efficiently transcribes early but not late herpes simplex virus type 1 mRNA. AB - The sequences of the DNAs encoding the 5' ends of one early and one late herpes simplex virus type 1 mRNA were analyzed, and the 5' ends of these mRNA species were precisely located. Neither mRNA species is spliced and the noncoding strand of the DNA contains recognizable T-A-T-A and C-A-T boxes upstream from their respective 5' ends. The early mRNA was efficiently transcribed by a commercially available uninfected cell lysate system, but the late mRNA was not. This difference between early and late mRNAs appears to be general in this virus. PMID- 6273853 TI - Carcinogen-mediated amplification of viral DNA sequences in simian virus 40 transformed Chinese hamster embryo cells. AB - Exposure of simian virus 40 (SV40)-transformed Chinese hamster embryo cells to various chemical and physical carcinogens induced SV40 DNA synthesis. Although the carcinogen-mediated amplification of SV40 DNA is regulated by the viral A gene, the induction of viral DNA synthesis does not result in the rescue of infectious virus or the formation of complete viral DNA molecules. Instead, a heterogeneous collection of DNA molecules containing SV40 sequences was generated by treatment with 7,12-dimethylbenz[a]anthracene. Restriction enzyme analysis of the amplified DNA molecules in the Hirt supernatant showed that not all sequences in the integrated SV40 inserts are present. The possibility that amplification of SV40 sequences is a reflection of a general-gene-amplification phenomenon mediated by carcinogens is discussed. PMID- 6273854 TI - Heteroduplex formation by recA protein: polarity of strand exchanges. AB - Purified recA protein promotes strand exchanges between linear duplex DNA and homologous circular single-stranded phage phi X174 DNA that carries a short hybridized fragment [West, S. C., Cassuto, E. & Howard-Flanders, P. (1981) Proc. Natl. Acad. Sci. USA 78, 2100-2104]. In this paper we investigate the mechanism of this strand exchange reaction. We show that recA protein initiates strand exchanges by pairing the free end of the duplex fragment with the single-stranded DNA. In addition, we find that strand exchanges are polar, stable heteroduplex molecules being formed by the directional transfer transfer of the (-) strands starting at 3' termini. PMID- 6273855 TI - Molecular complexes involved in the regulation of adenylate cyclase. AB - Selective extraction of the adenylate cyclase regulatory protein (N-protein) from pigeon erythrocyte plasma membranes provided evidence for its cytoskeletal association. Cholate, but not Triton X-100 or digitonin, was effective in solubilizing the ADP-ribosylated N-protein. The labeled protein complex or components thereof that were associated with the Triton-insoluble cytoskeleton (shells) could be partly released by 0.1 mM EDTA; 1 M KCl in the presence of Triton X-100 achieved complete solubilization. 5'-Guanylyl imidodiphosphate (p[NH]ppG) and NaF, activators of adenylate cyclase, promoted the release of the regulatory protein from the cytoskeleton but MnCl2, an "uncoupler" of the adenylate cyclase system, had the opposite effect. The solubilized, labeled N protein was able to bind specifically to rat erythrocyte inside-out vesicles in the presence of divalent cations. A proteolytic product of inside-out vesicles inhibited the binding of the N-protein to fresh vesicles. Three molecular species which contained the Mr 45,000 polypeptide component of the N-protein were identified by gel permeation chromatography and by sucrose density gradient velocity sedimentation. p[NH]ppG appeared to convert the two larger molecular complexes to a smaller molecular entity. Such a molecular dissociation might be relevant to the effects of guanyl nucleotides on the activity of adenylate cyclase and on the affinity of hormone receptors. PMID- 6273856 TI - Preparation of tritium- or deuterium-labeled vitamin D analogs by a convenient general method. AB - The three-step conversion of vitamin D analogs to 6-oxo-3,5-cyclovitamin D derivatives followed by reduction with a tritide or deuteride reagent and subsequent cycloreversion gives 6-tritio(deutero)vitamin D derivatives and corresponding 5,6-trans-analogs. The method is general and affords the 6-labeled vitamin D analogs in approximately 20% overall yield. PMID- 6273857 TI - Classification of multiple morphine and enkephalin binding sites in the central nervous system. AB - Detailed competitive displacement curves of 3H-labeled [D Ala2,Met5]enkephalinamide, [D-Ala2,D-Leu5]enkephalin, and dihydromorphine by a series of opiates and enkephalins are biphasic, suggesting multiple sites. After treatment of tissue with naloxazone, the displacement of the three 3H-labeled ligands by all opiates and enkephalins tested becomes monophasic, losing the high affinity displacement seen with low concentrations of both opiates and enkephalins. Coupled with Scatchard analysis of saturation experiments, these findings suggest a common site that binds both opiates and enkephalins equally well and with highest affinity (Kd values, less than 1 nM). Termed the mu 1 site, it corresponds to the previously described high-affinity site and appears to be the site responsible for analgesia under normal circumstances. The low-affinity binding of [3H]dihydromorphine (Kd, 3 nM) remaining after naloxazone treatment differs dramatically from low-affinity [D-Ala2,D-Leu5]-[3H]enkephalin binding (Kd, 5 nM). The mu 2 site, corresponding to the low-affinity [3H]dihydromorphine receptor sites, binds morphine (Ki, 10 nM) and dihydromorphine (Kd, 3 nM) far better than [D-Ala2,D-Leu5]enkephalin (Ki, 50 nM). Low-affinity [D-Ala2,D-Leu5] [3H]enkephalin receptor sites bind [D-Ala2,D-Leu5]enkephalin (Ki, 5-8 nM) more potently than morphine (Ki, 71 nM) and correspond to the previously established delta receptor. PMID- 6273858 TI - Sequence-specific contacts between the RNA polymerase of vesicular stomatitis virus and the leader RNA gene. AB - Methylation-protection studies with nucleocapsids from vesicular stomatitis virus indicate that the viral polymerase (L and NS proteins) contacts the genomic RNA template in the middle of the leader gene, 16-30 nucleotides from the 3' terminus. The data suggest that the NS protein binds to the sequence (formula, see text) and may function as an initiator protein for transcription. PMID- 6273859 TI - Coordination environment of the active-site metal ion of liver alcohol dehydrogenase. AB - The coordination environment of the catalytically active metal ion of horse liver alcohol dehydrogenase (alcohol:NAD+ oxidoreductase, EC 1.1.1.1) has been investigated by electron paramagnetic resonance (EPR) methods with use of the active-site-specific Co2+-reconstituted enzyme. The EPR absorption spectrum of the metal-substituted enzyme is characteristic of a rhombically distorted environment. The spectrum of the enzyme--NAD+ complex shows approximate axial symmetry of the metal ion site, indicating that binding of the coenzyme induces a structural alteration in the active-site region. This environment is not significantly altered further by binding of the competitive inhibitor pyrazole. To assign the coordination number of the active-site metal ion, the zero-field splitting was determined on the basis of the temperature dependence of the spin- lattice relaxation of the Co2+ ion. The zero-field splitting energies are approximately 9 cm-1 for the free Co2+-reconstituted enzyme and approximately 46 and approximately 47 cm-1 for the enzyme--NAD+ and enzyme--NAD+--pyrazole complex, respectively. On the basis of studies of structurally defined small molecule complexes, these values are compatible with a tetracoordinate metal ion in the active site of the free enzyme but a pentacoordinate metal ion in the binary enzyme--NAD+ complex and in the ternary enzyme--NAD+--inhibitor complex and, therefore, presumably also in the catalytically active ternary enzyme--NAD+- alcohol complex formed in the course of alcohol oxidation. PMID- 6273860 TI - Metabolism of benzo[a]pyrene by human mammary epithelial cells: toxicity and DNA adduct formation. AB - Pure cultures of human breast epithelial cells and of fibroblastic cells in early passage provided the opportunity to ask whether either cell type had the capability for metabolizing chemical carcinogens and, if so, was the fate of the metabolic products compatible with chemical carcinogens being a factor in the initiation of breast cancer in women. For this purpose, cells were exposed to benzo[a]pyrene (BaP), and (i) the influence on growth potential and (ii) the extent, type, and persistence of adducts between the metabolites of BaP and DNA were measured. Compared with fibroblasts, inhibition of growth by epithelial cells was 50-100 times more sensitive to BaP. Because of this differential sensitivity, epithelial cells were exposed to 0.4 microM BaP and fibroblasts were exposed to 4.0 microM BaP in the studies of DNA adduct formation. Separation by high-pressure liquid chromatography of adducts between (+/-)-7,8-dihydroxy-9,10 epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene (BaP diol epoxide) and nucleosides from purified DNA revealed that epithelial cells contained modified DNA within 6 hr after adding BaP. Adducts between the 7R anti stereoisomer of BaP diol epoxide and deoxyguanosine predominated at all times. syn BaP diol epoxide adducts with deoxyguanosine and what appeared to be BaP diol epoxide adducts with deoxycytidine were consistently present but at much lower frequency. All three types of BaP diol epoxide--DNA adducts persisted in epithelial cells for 72 hr in BaP-free medium. No adducts were detected in fibroblastic cultures until 96 hr after first exposure to BaP. At this time, the type and extent of BaP diol epoxide--DNA adduct formation was similar to that in epithelial cells exposed to one-tenth the dose of BaP. The type, extent, rate of formation, and persistence of the adducts in human breast epithelial cells was similar to that in cells transformable by exposure to BaP, an indication that they may be targets for chemically induced carcinogenesis. PMID- 6273861 TI - Mapping functional domains in the promoter region of the herpes thymidine kinase gene. AB - The cloned herpes simplex virus type 1 (HSV-1) thymidine kinase (TK; ATP:thymidine 5'-phosphotransferase, EC 2.7.1.21) gene can be used to transform TK- cells to a TK+ phenotype. Transformants generated in this way express TK at a basal constitutive level that is inducible to a higher level by infection with TK herpes virus. We have studied the effect of mutations generated in vitro on both the constitutive and virus-induced expression of TK in transformants. Four Xho I linker insertions and two deletions in the 5' untranscribed region of the cloned HSV-1 TK gene were generated in vitro. A deletion that removed all but nine base pairs of the 5' untranscribed region virtually eliminated constitutive expression and completely prevented induction by herpes virus infection. Two of the insertions have particularly interesting properties. One, nine base pairs upstream from the cap site, inactivates constitutive expression without stopping induction. The other, 50 base pairs upstream from the cap site has the opposite effect (i.e., normal constitutive expression but no induction). Analysis of these results leads us to propose that the 5' untranscribed region of the HSV-1 TK gene is quite complex with several functional domains having differential roles in the constitutive and herpes-induced expression of the TK gene. PMID- 6273862 TI - Desensitization to gonadotropins in cultured Leydig tumor cells involves loss of gonadotropin receptors and decreased capacity for steroidogenesis. AB - The ability of human choriogonadotropin (hCG) to regulate its receptors and target cell responses has been studied in a clonal strain of cultured Leydig tumor cells (MA-10). Exposure of the MA-10 cells to hCG results in decrease in hCG binding activity which is dependent on time and the concentration of hCG. This decrease is due to a change in the number of receptors rather than in the affinity of the receptors, and it is accompanied by a corresponding reduction in the ability of hCG to stimulate steroidogenesis. Exposure of the MA-10 cells to hCG also resulted in a reduction of the steroidogenic responses to cholera toxin and 8-Br-adenosine cyclic 3',5'-monophosphate. The hCG-induced loss of steroidogenic responses to these stimuli seems to be due to the stimulation of steroidogenesis rather than to the decrease in hCG receptors because it also can be induced when steroidogenesis is stimulated with cholera toxin or 8-Br adenosine 3',5'-monophosphate under conditions such that the number of hCG receptors is not reduced. PMID- 6273863 TI - Extensive polymorphism in the mitochondrial DNA of apes. AB - Ape species are 2-10 times more variable than the human species with respect to the nucleotide sequence of mtDNA, even though ape populations have been smaller than the human population for at least 10,000 years. This finding was made by comparing purified mtDNAs from 27 individuals with the aid of 25 restriction endonucleases; for an additional 59 individuals, comparisons were made with fewer enzymes by using the blot hybridization method. The amount of intraspecific sequence divergence was greatest between orangutans of Borneo and Sumatra. Among common chimpanzees, a large component of the variation is due to two highly distinct forms of mtDNA that may reflect a major geographic subdivision. The least amount of sequence variation occurred among lowland gorillas, which exhibit only twice as much sequence variation as humans. The large intraspecific differences among apes, together with the geological and protein evidence, leads us to propose that each ape species is the remnant of an ancient and widespread population that became subdivided geographically and reduced in size and range, perhaps by hominid competition. The low variation among human mtDNAs is consistent with geological evidence that the human species is young. The distribution of site changes within the mitochondrial genome was also examined. Comparison of closely related mtDNAs shows that the ribosomal RNA genes have diverged more slowly than the rest of the genome. PMID- 6273864 TI - Estimating genetic divergence and genetic variability with restriction endonucleases. AB - Restriction endonucleases cut DNA at specific sites determined by the local nucleotide sequence. By comparing related DNA segments with respect to where such cuts are made, one can estimate the extent of sequence homology between the segments. Empirical methods are presented here for using these data to measure the proportion of mismatches between two sequences, the proportion of polymorphic positions in a series of sequences, or the degree of heterozygosity in a population. These methods do not require any assumptions concerning the evolutionary or population genetic processes involved. One can also use the data to calculate the precision of each of these estimates. When the positions of the cuts are not determined, these estimates can be made, using only the lengths of the resulting DNA fragments, by means of a maximum likelihood procedure. Several examples demonstrate the usefulness of these methods to study genetic differences in regions of the genome not amenable to study by other methods. PMID- 6273865 TI - The cyc1-11 mutation in yeast reverts by recombination with a nonallelic gene: composite genes determining the iso-cytochromes c. AB - DNA sequence analysis of a cloned fragment directly established that the cyc1-11 mutation of iso-1-cytochrome c in the yeast Saccharomyces cerevisiae is a two base-pair substitution that changes the CCA proline codon at amino acid position 76 to a UAA nonsense codon. Analysis of 11 revertant proteins and one cloned revertant gene showed that reversion of the cyc1-11 mutation can occur in three ways: a single base-pair substitution, which produces a serine replacement at position 76; recombination with the nonallelic CYC7 gene of iso-2-cytochrome c, which causes replacement of a segment in the cyc1-11 gene by the corresponding segment of the CYC7 gene; and either a two-base-pair substitution or recombination with the CYC7 gene, which causes the formation of the normal iso-1 cytochrome c sequence. These results demonstrate the occurrence of low frequencies of recombination between nonallelic genes having extensive but not complete homology. The formation of composite genes that share sequences from nonallelic genes may be an evolutionary mechanism for producing protein diversities and for maintaining identical sequences at different loci. PMID- 6273866 TI - Yeast transformation: a model system for the study of recombination. AB - DNA molecules that integrate into yeast chromosomes during yeast transformation do so by homologous recombination. We have studied the way in which circular and linear molecules recombine with homologous chromosomal sequences. We show that DNA ends are highly recombinogenic and interact directly with homologous sequences. Circular hybrid plasmids can integrate by a single reciprocal crossover, but only at a low frequency. Restriction enzyme digestion within a region homologous to yeast chromosomal DNA greatly enhances the efficiency of integration. Furthermore, if two restriction cuts are made within the same homologous sequence, thereby removing an internal segment of DNA, the resulting deleted-linear molecules are still able to transform at a high frequency. Surprisingly, the integration of these gapped-linear molecules results in replacement of the missing segment using chromosomal information. The final structure is identical to that obtained from integration of a circular molecule. The integration of linear and gapped-linear molecules, but not of circular molecules, is blocked by the rad52-1 mutation. Consideration of models for plasmid integration and gene conversion suggests that RAD52 may be involved in the DNA repair synthesis required for these processes. Implications of this work for the isolation of integrative transformants, fine-structure mapping, and the cloning of mutations are discussed. PMID- 6273867 TI - Molecular basis for familial isolated growth hormone deficiency. AB - Nuclear DNA from four individuals with familial isolated growth hormone (somatotropin) deficiency (IGHD) type A was studied by restriction endonuclease analysis. By using 32P-labeled human growth hormone (hGH) cDNA sequences as a probe, patterns seen after various digestions indicated that these individuals were homozygous for a deletion of at least 7.5 kilobases (kb) of DNA. This deletion includes the gene that encodes the normal growth hormone but does not include the variant growth hormone gene. Restriction patterns of DNAs from all family members agreed with an autosomal recessive mode of inheritance of the deletion that correlates with the clinical phenotype. Furthermore, independent assortment of the two types of hGH genes suggests that these genes are nonallelic. These findings indicate that, in these families, IGHD type A is caused by deletion of the normal hGH genes and that this disorder can occur in the presence of variant hGH genes. PMID- 6273868 TI - Transcription of the simian virus 40 genome in DNA-transformed murine teratocarcinoma stem cells. AB - To study the molecular basis for lack of expression of the simian virus 40 (SV40) early region genes in murine teratocarcinoma-derived stem cells, we introduced a recombinant plasmid consisting of pBR322 linked to the herpes simplex virus type 1 thymidine kinase gene and SV40 genome into thymidine kinase-deficient F9 stem cells. The resulting stem cell clone, 12-1, and a retinoic acid-induced differentiated daughter cell clone, 12-1a, each contain one copy per cell of the entire recombinant plasmid integrated into the cellular genome through a site on the pBR322 genome. Restriction endonuclease analyses indicate that there is no difference in integration site or organization of the three component parts of the plasmid genome within cellular DNA of stem and differentiated cells; yet the differentiated cells, 12-1a, express SV40 large tumor antigen whereas the stem cells, 12-1, do not. Both stem and differentiated cells produce two size classes of polyadenylylated RNA, 2900 and 2600 bases in length, homologous to the early region of the SV40 genome, detectable by RNA blotting analysis. S1 nuclease analysis of the SV40 transcripts present in stem and differentiated cells indicate that the SV40 mRNAs were identically spliced in the two cell types, in a manner consistent with that observed for spliced large and small tumor antigen mRNAs in SV40-infected monkey kidney cells. Thus, the failure of 12-1 teratocarcinoma stem cells, containing an integrated SV40 genome, to express SV40 tumor antigen is not due to a lack of transcription of the SV40 early region or to an inability to splice primary transcripts. PMID- 6273869 TI - Herpesvirus ateles and herpesvirus saimiri transform marmoset T cells into continuously proliferating cell lines that can mediate natural killer cell-like cytotoxicity. AB - Herpesvirus ateles (HVA) and herpesvirus saimiri (HVS) have the capacity to transform cotton-topped marmoset T lymphocytes into continuously proliferating cell lines that retain some functions associated with cell-mediated immunity. In the present paper, we demonstrate that HVA/HVS-transformed T cell lines are cytotoxic in a short-term 51Cr release assay and that this killing resembles killing by marmoset natural killer (NK) cells. The relationship between NK cells and HVA/HVS-transformed killer cell lines is discussed in view of present knowledge of the origin and function of the NK system. It is suggested that the described cytotoxic cell lines may be useful for further defining cellular cytotoxicity with regard to cell surface recognition, regulatory events, and lytic mechanisms. PMID- 6273870 TI - Single channel currents induced by complement in antibody-coated cell membranes. AB - An extracellular patch electrode was used to record ionic currents from individual complement-induced channels in the membranes of antibody-coated skeletal muscle. The amplitude of the single-channel currents leads to an estimate of 90 pS for the unit conductance. The kinetics of channel opening and closing show marked variability and complexity. Channels flicker open and closed repeatedly, indicating that once these lesions form, they undergo rapid structural transitions between discrete conducting and nonconducting states. PMID- 6273871 TI - Episomal simian virus 40 genomes in human brain tumors. AB - Eight out of 35 human intracranial tumors were shown by restriction enzyme analysis to contain unintegrated simian virus 40 (SV40) DNA molecules. The relative amount of viral DNA was estimated to be the equivalent of one viral genome within every 10th to 20th cell. No infectious virus was detected in tissue cultures established from the tumors. From only one tumor was it possible to rescue, by cell fusion, infectious SV40 displaying wild-type properties. In those cases that permitted a more detailed analysis, the restriction enzyme cleavage patterns appeared to correspond to the wild-type patterns with one exception, in which the SV40 episomes displayed a deletion of approximately 70 base pairs close to the origin of DNA replication. From one tumor, the SV40 genomes were transferred into permissive CV-1 monkey cells by transfection with the total tumor DNA. Despite their persistence as episomes no infectious virus was produced. Furthermore, no viral antigens were detectable, although the SV40 messengers for the small and the large tumor antigens were present. These cells had, however, acquired the ability to form colonies in low concentrations of serum. Thus this report provides, by restriction enzyme analysis, direct evidence for the presence of SV40 DNA in human tumors. PMID- 6273873 TI - Enhanced macrophage degradation of low density lipoprotein previously incubated with cultured endothelial cells: recognition by receptors for acetylated low density lipoproteins. AB - Human low density lipoprotein (LDL) was incubated with an established line of rabbit aortic endothelial cells. Density gradient fractionation showed a time-, concentration-, and temperature-dependent increase in the average density of the LDL (from about 1.036 to as high as 1.070 g/ml). Incubation without cells or with other types of cultured cells (fibroblasts, hepatocytes, 3T3-L1 cells) caused no significant change in density. 125I-Labeled LDL (125I-LDL) recovered after incubation with endothelial cells (EC-modified LDL) was taken up and degraded 3 to 4 times more rapidly than control LDL by resident mouse peritoneal macrophages and by an established tumor line of mouse macrophages (J774 cells). Macrophage degradation of EC-modified 125I-LDL exhibited saturation kinetics (greater than 85% inhibited by excess unlabeled EC-modified LDL). Degradation was also inhibited by unlabeled acetylated LDL and, conversely, unlabeled EC-modified LDL inhibited degradation of acetylated 125I-LDL. Incubation of LDL with conditioned medium-removed from endothelial cell cultures modified neither its density nor its rate of degradation by macrophages. These studies show that endothelial cells have the potential to metabolically modify the LDL molecule, generating a form that is more rapidly degraded by macrophages and that is recognized by the macrophage receptor for acetylated LDL. This process may play a significant role in the pathogenesis of atherosclerosis. PMID- 6273872 TI - Herpes simplex type 1 DNA in human brain tissue. AB - Herpes simplex virus type 1 (HSV-1) is known to reside latently in the trigeminal ganglia of man. Reactivation of this virus causes skin lesions and may occasionally infect other tissues, including the brain. To determine whether the brain tissue of humans free of clinical signs of HSV-1 infection contains any trace of HSV-1, we examined the DNA from brain tissue by endonuclease digestion, separation of the fragments by gel electrophoresis, and hybridization with labeled HSV-1 DNA probes. Hybrid bands were detected autoradiographically in experiments using cloned and virion-purified fragments of the HSV-1 genome. HSV-1 DNA sequences were found in 6 of 11 human brain DNA samples tested. In some cases, these bands corresponded to the bands expected for the complete viral genome, whereas others contained bands representing only a part of the genome. In some cases, the terminal fragments could be found, suggesting that the DNA was in a linear, nonintegrated form. PMID- 6273874 TI - Primary structure of corticotropin-releasing factor from ovine hypothalamus. AB - Sequence analysis was performed of an ovine hypothalamic 41-residue polypeptide that had been postulated to be a putative corticotropin-releasing factor (CRF) because of its high intrinsic corticotropin releasing activity. The NH2-terminal 39 residues of CRF were determined by Edman degradation of 0.6-3.5 nmol of peptide in a Wittmann-Liebold modified Beckman 890C spinning cup sequencer with reverse-phase high-pressure liquid chromatography for the identification of amino acid phenylthiohydantoins (direct micro-sequence analysis). Evidence for residue 40 (isoleucine) was provided by direct micro-sequence analysis of 2.0 nmol of acetylated CRF selectively cleaved at its arginine residues by trypsin prior to analysis. The thermolytic COOH-terminal fragment isoleucyl-alanineamide was characterized as its dansyl derivative. Based on the analytical data, the following primary structure is proposed for ovine hypothalamic CRF: H-Ser-Gln-Glu Pro-Pro-Ile-Ser-Leu-Asp-Leu-Thr-Phe-His-Leu-Leu-Arg-Glu-Val-Leu-Glu-Met-Thr-Lys Ala-Asp-Gln-Leu-Ala-Gln-Gln-Ala-His-Ser-Asn-Arg-Lys-Leu-Leu-Asp -Ile-Ala-NH2. In agreement with this proposal, the synthetic replicate of CRF is highly potent in stimulating secretion of both corticotropin and beta-endorphin-like immunoactivities. PMID- 6273875 TI - Repression and activation of the genome of herpes simplex viruses in human cells. AB - We have described previously a cell culture system in which the herpes simplex virus (HSV) type 2 (HSV-2) genome is maintained in a repressed form after treatment of infected cells with 1-beta-D-arabinofuranosylcytosine and increase of incubation temperature from 37 degrees C to 39.5 degrees C. Infectious HSV-2 production was activated by altering incubation temperature or by superinfecting with human cytomegalovirus. We now report the establishment of an analogous system utilizing HSV type 1 (HSV-1). Human embryo lung cells were infected with HSV-1 and treated with 1-beta-D-arabinofuranosylcytosine (25 micrograms/ml) for 7 days to minimize both synthesis of virus DNA and infectious virus while allowing expression of early virus genes. HSV-1 was maintained in an undetectable form for at least 72 days when the incubation temperature was raised from 37 degrees C to 40.5 degrees C after removal of the inhibitor. HSV-1 gene expression was then predictably turned on by superinfection with human cytomegalovirus or by reducing the incubation temperature. Virus replicated after activation was compared with the respective parental virus with regard to inhibition by the HSV-1-specific antiviral (E)-5-(2-bromovinyl)-2'-deoxyuridine and EcoRI, HindIII, and Xba I restriction endonuclease cleavage patterns. The results show activation of HSV gene expression in human cells by a human cytomegalovirus early gene function(s), followed by synthesis of parental-like HSV. PMID- 6273876 TI - Purine metabolism in the protozoan parasite Eimeria tenella. AB - Crude extracts of the oocysts of Eimeria tenella, a protozoan parasite of the coccidium family that develops inside the caecal epithelial cells of infected chickens, do not incorporate glycine or formate into purine nucleotides; this suggests lack of capability for de novo purine synthesis by the parasite. The extracts, however, contain high levels of activity of the purine salvage enzymes: hypoxanthine, guanine, xanthine, and adenine phosphoribosyltransferases and adenosine kinase. The absence of AMP deaminase from the parasite indicates that E. tenella cannot convert AMP to GMP; the latter thus has to be supplied by the hypoxanthine, xanthine, or guanine phosphoribosyltransferase of the parasite. These three activities are associated with one enzyme (HXGPRTase), which has been purified to near homogeneity in high yield (71-80%) in a single step by GMP agarose affinity column chromatography. The size of the enzyme subunit is estimated to be 23,000 daltons by NaDodSO4 gel electrophoresis. Kinetic studies suggest differences in purine substrate specificity between E. tenella HXGPRTase and chicken liver HGPRTase. Allopurinol preferentially inhibits the parasite enzyme by competing with hypoxanthine; a Ki approximately 22 microM. PMID- 6273877 TI - Purified glucocorticoid receptors bind selectively in vitro to a cloned DNA fragment whose transcription is regulated by glucocorticoids in vivo. AB - Activated glucocorticoid receptor protein, purified to 40-60% homogeneity from rat liver extracts, binds selectively in vitro to a cloned fragment of murine mammary tumor virus (MTV) DNA. The DNA fragment tested contains about half of the sequences present in intact MTV DNA, and its rate of transcription, like that of the intact viral element, is strongly stimulated by glucocorticoids when it is introduced into the genome of a receptor-containing cell. In contrast, the receptor fails to bind selectively to DNA restriction fragments from E. coli plasmids pBR322 and RSF2124 or from bacteriophages lambda and T4. Preliminary experiments to localize regions within MTV DNA responsible for selective binding have revealed thus far one subfragment that fails to bind the receptor and one selectively bound subfragment that maps far downstream from the 5' terminus of the normal RNA transcript. These studies are consistent with the notion that steroid receptors may modulate rates of transcription by recognizing specific DNA sequences within or near the regulated genes. PMID- 6273878 TI - Enzymatic synthesis of biotin-labeled polynucleotides: novel nucleic acid affinity probes. AB - Analogs of dUTP and UTP that contain a biotin molecule covalently bound to the C 5 position of the pyrimidine ring through an allylamine linker arm have been synthesized. These biotin-labeled nucleotides are efficient substrates for a variety of DNA and RNA polymerases in vitro. Polynucleotides containing low levels of biotin substitution (50 molecules or fewer per kilobase) have denaturation, reassociation, and hybridization characteristics similar to those of unsubstituted controls. Biotin-labeled polynucleotides, both single and double stranded, are selectively and quantitatively retained on avidin-Sepharose, even after extensive washing with 8 M urea, 6 M guanidine hydrochloride, or 99% formamide. In addition, biotin-labeled polynucleotides can be selectively immunoprecipitated in the presence of antibiotin antibody and Staphylococcus aureus protein A. The unique features of biotin-labeled polynucleotides suggest that they will be useful affinity probes for the detection and isolation of specific DNA and RNA sequences. PMID- 6273879 TI - Fate of viral DNA in nonpermissive cells infected with simian virus 40. AB - Mouse cells are nonpermissive for simian virus 40 (SV40); replication of viral DNA is undetectable and progeny virions are not produced. Infection leads instead to the establishment of stably transformed cell lines in which viral DNA is covalently integrated into cellular DNA. We have followed the fate of SV40 DNA in infected mouse cells to define steps in viral DNA metabolism that precede integration. A novel high molecular weight form of SV40 DNA is synthesized shortly after infection by a process sensitive to the inhibition of DNA replication. This DNA represents polymers in which viral genomes are organized as tandem "head-to-tail" arrays. Recombination can be demonstrated with mutant viruses, but the recombination frequency is not high enough to account for the synthesis of polymers by recombination between infecting genomes. We conclude that polymers are synthesized by DNA replication and that they then recombine with one another. We believe that the polymers also recombine with cellular DNA and are thus the precursor to integrated viral DNA. Such a model accounts directly for the high frequency of tandemly duplicated viral insertions in transformed cells and also leads to experimentally testable predictions. PMID- 6273880 TI - The dicyclohexylcarbodiimide-binding protein c of ATP synthase from Escherichia coli is not sufficient to express an efficient H+ conduction. AB - Bacteriophage Mu was inserted into the unc genes of Escherichia coli. The resulting mutation AS12 had a polar effect on the unc operon: membranes of the mutant AS12 contained the dicyclohexylcarbodiimide-binding protein c and the protein a as sole subunits of the ATP synthase. It was shown by peptide mapping and amino acid analysis of the fragments that protein c from mutant AS12 was identical with the wild-type protein c. The absence of subunit b in mutant AS12 drastically lowered the H+ conduction dependent on the membrane-integrated moiety (F0) of the ATP synthase. This suggests that both subunits b and c are necessary for an efficient expression of H+ conduction. PMID- 6273881 TI - Conditional expression of the vesicular stomatitis virus glycoprotein gene in Escherichia coli. AB - Bacterial plasmids that directed expression of the vesicular stomatitis virus glycoprotein (G-protein) gene under control of the tryptophan operon regulatory region were constructed. A plasmid directing the synthesis of a G-protein-like protein (containing the NH2-terminal segment of seven amino acids encoded by the trpE gene fused to the complete G-protein sequence lacking only its NH2-terminal methionine) could be transformed into trpR+ (repressed) but not into trpR- (derepressed) cells. This result suggested initially that derepressed synthesis of the G-protein-like protein encoded by this plasmid was lethal in Escherichia coli. Deletion of the sequence encoding the large hydrophobic segment near the COOH terminus of G-protein did not overcome this lethality. Lethality of derepressed synthesis was overcome by deletion of the G-protein gene region encoding 10 amino acids in the hydrophobic NH2-terminal domain (signal peptide). Tryptic peptide mapping demonstrated that the G-protein-like protein and some truncated proteins encoded by the plasmid contained G-protein protein sequences. Antisera to vesicular stomatitis virus precipitated the G-protein-like protein, showing that it shares antigenic determinants with the authentic G-protein protein. PMID- 6273883 TI - Molecular cloning of equine herpesvirus type 1 DNA: analysis of standard and defective viral genomes and viral sequences in oncogenically transformed cells. AB - Genomic DNA sequences of equine herpesvirus type 1 (EHV-1) have been cloned as BamHI and EcoRI restriction fragments into the plasmid pBR322 and propagated in Escherichia coli. With the exception of two EcoRI restriction fragments that reside in the S region of the viral genome, all of the cloned fragments demonstrated the same electrophoretic mobilities, restriction cleavage sites, and blot-hybridization patterns as did the parent fragments produced by BamHI or EcoRI digestion of virion DNA. The EcoRI J fragment and the BamHI E fragment of the L-region terminus were cloned after the addition of appropriate linker oligonucleotides. Fragments originating from each of the two isomeric forms of EHV-1 DNA were contained in this library of clones. Supramolar DNA fragments present only in the DNA of defective interfering (DI) particles of EHV-1 were generated from Bgl II digestion of DNA preparations enriched for EHV-1 DI particles and were cloned as Bgl II and EcoRI fragments into the plasmid vector. The cloned viral sequences represented in this defective genome mapped to the S region of EHV-1 DNA. Blot-hybridization analysis of EHV-1 transformed and tumor cell DNAs with the cloned BamHI B fragment confirmed that subgenomic viral sequences are present and indicated that those sequences map to the viral genome between 0.32 and 0.43 map unit. PMID- 6273882 TI - Scrapie agent contains a hydrophobic protein. AB - The scrapie agent causes a degenerative nervous system disorder of sheep and goats. Considerable evidence indicates that the scrapie agent contains a protein that is necessary for infectivity [Prusiner, S. B., Groth, D. F., Cochran, S. P., Masiarz, F. R., McKinley, M. P. & Martinez, H. M. (1980) Biochemistry 19, 4883 4891], but direct demonstration of a protein moiety has been hampered by lack of sufficiently purified preparations. Employing preparations of the scrapie agent enriched 100- to 1000-fold with respect to protein, we found that digestion by proteinase K destroyed more than 99.9% of the infectivity. Diethylpyrocarbonate, which chemically modifies amino acid residues in proteins with high efficiency, also inactivated the scrapie agent in these purified preparations. Reductions of infectivity by proteinase K and diethylpyrocarbonate were not observed with less purified preparations. The agent bound to phenyl-Sepharose could not be eluted with 8.5 M ethylene glycol; however, a combination of ethylene glycol and detergents did release the agent. These observations provide good evidence for a protein and for hydrophobic domains within the scrapie agent. Whether the protein required for infectivity is the same protein responsible for the hydrophobic properties of the scrapie agent remains to be established. PMID- 6273885 TI - Metal thiolate clusters in cobalt(II)-metallothionein. AB - Rabbit liver metallothionein-1 in which all seven metal-binding sites are occupied by cobalt(II) exhibits spectral features typical of tetrathiolate coordination with approximate Td microsymmetry [Vasak, M. (1980) J. Am. Chem. Soc. 102, 3953-3955]. With a total of 20 cysteine residues per molecule, this mode of metal binding implies that some of the thiolate ligands are shared by neighboring Co(II) ions, resulting in clustered structures. In this study, evidence for the existence of thiolate-linked Co(II) clusters is presented and their mode of formation is explored by comparing the optical and magnetic properties of forms of Co(II)-metallothionein containing 1-7 equivalents of Co(II). Preparations with up to 4 Co(II) equivalents display electronic spectra in the d-d and charge-transfer regions that resemble those of isolated tetrahedral Co(II)-tetrathiolate complexes. Upon binding of more than four Co(II) ions, however, the spectrum changes progressively and approaches in the fully saturated Co(II)-metallothionein an absorption profile similar to that of crystallographically defined model (Co)II-tetrathiolate clusters [Dance, I. G. (1979) J. Am. Chem. Soc. 101, 6264-6273]. These effects are closely paralleled by changes in the ESR spectrum. Above 4 Co(II) equivalents per thionein, the ESR signal at gx approximately 5.9 measured at 4 K decreases progressively in intensity, until in the fully occupied protein the complex is nearly diamagnetic. These changes, which were confirmed by measurements of paramagnetic susceptibility, establish the existence of Co(II) thiolate clusters in Co(II) metallothionein. The loss of paramagnetism reflects most likely antiferromagnetic coupling of neighboring Co(II) ions brought about by a superexchange mechanism via the thiolate bridging ligands. PMID- 6273884 TI - Tyrosylprotein kinase and phosphatase activities in membrane vesicles from normal and Rous sarcoma virus-transformed rat cells. AB - Membrane vesicles; isolated from normal and Rous sarcoma virus-transformed rat cells, have an associated cyclic-AMP independent kinase that phosphorylates a Mr 37,000 protein in vesicles from normal cells and proteins of Mr 37,000, 50,000, and 67,000 in vesicles from transformed cells. Proteins in vesicles from normal and transformed cells contain 9% and 77%, respectively, of their labeled phospho amino acids as phosphotyrosine. Thus, isolation of vesicles and subsequent incubation with [gamma-32P]ATP enriches the proportion of labeled phosphotyrosine in proteins (relative to other phospho amino acids) by two orders of magnitude over that found in intact cells. The in vitro phosphorylation of each of these proteins is enhanced in the presence of 10 microM Zn2+, a phosphotyrosylprotein phosphatase inhibitor. From these studies it appears that membrane vesicles may be a valuable system for examination of transformation-specific phosphorylation of proteins. PMID- 6273886 TI - Laser flash photolysis studies of electron transfer between semiquinone and fully reduced free flavins and horse heart cytochrome c. AB - Laser flash photolysis has been used to determine the second-order rate constants for the reduction of horse heart cytochrome c by the semiquinone and fully reduced forms of various flavin analogs. We find that substitution in the dimethylbenzene ring of the flavin causes appreciable changes in the rate constants, whereas substitutions at the N-10 position do not. Placing a charged phosphate group in the N-10 ribityl side chain leads to only small ionic strength effects on the rate constants, whereas a charged group attached to the dimethylbenzene ring produces a large ionic strength effect. These results can be accounted for by assuming that a productive collision between flavin and cytochrome involves an orientation that positions the aromatic ring--N-5 region of the flavin toward the heme crevice and the N-10--pyrimidine ring region away from it. Our observations have implications for mechanistic understanding of biological electron transfer reactions and are discussed in this context. PMID- 6273887 TI - Human beta-endorphin: specific binding in neuroblastoma N18TG2 cells. AB - The mouse neuroblastoma N18TG2 has specific, saturable binding sites for human beta-endorphin (beta h-EP). The affinity and number of sites are 1.1 nM and 280,000 per cell, respectively, beta h-EP binding is not inhibited by [Leu]enkephalin or morphine at concentrations up to 0.1 mM; beta h-EP-(6--31) is a potent inhibitor of binding, and camel beta-EP is much less potent. The data suggest the importance of the nonenkephalin segment of the beta h-EP molecule for interaction with the binding site in N18TG2 cells. PMID- 6273888 TI - Viable deletions of the M13 complementary strand origin. AB - The single-stranded DNA of bacteriophage M13 is converted to a duplex replicative form by a mechanism involving RNA-primed initiation at a single unique site on the viral DNA. The DNA sequence that specifies the RNA primer is contained largely within one of two adjacent hairpin structures protected from DNase degradation by RNA polymerase. We have used in vitro techniques to construct a series of M13 mutants having deletions in the region of the complementary strand origin. Deletions of the duplex replicative form DNA range in size from 54 to 201 base pairs. The largest deletions remove both of the RNA polymerase-protected hairpins and the entire sequence specifying the primer RNA. Mutants lacking one or both hairpins form faint plaques, give reduced phage yields, and show a lag in phage production of >30 min. The rate of conversion of the single-stranded viral DNA to the parental replicative form is reduced both in vivo and in vitro. These results indicate that both the RNA polymerase-protected hairpins and the RNA primer-coding sequence are important, but not essential, for replication. Other sequences within the origin region, or possibly elsewhere in the genome, may play a role in complementary strand initiation in these mutant phages. The M13 viral strand is initiated by extension of the 3' terminus generated by site-specific nicking of the viral strand of the replicative form DNA by the M13 gene II protein. This specific nicking site is retained in all of the M13 deletion mutants. Deletion end points do not extend into a 13-nucleotide sequence preceding the viral strand nicking site. We propose that a sequence including these 13 nucleotides is required for gene II protein action at this site. PMID- 6273889 TI - Bridging of IgE receptors activates phospholipid methylation and adenylate cyclase in mast cell plasma membranes. AB - Bridging of IgE receptors on normal rat mast cells by divalent anti-receptor antibodies induced phospholipid methylation and an increase in intracellular cyclic AMP within 15 sec after the receptor bridging. These biochemical events were followed by Ca2+ influx and histamine release. When IgE receptors on isolated plasma membranes were bridged by the antibody, both the increase in the incorporation of [3H]methyl into lipid fraction and the synthesis of cyclic AMP were demonstrated. The synthesis of cyclic AMP in this system was enhanced in the presence of GTP. The results indicated that the bridged IgE receptors are linked to both methyltransferases and adenylate cyclase [ATP pyrophosphate-lyase (cyclizing), EC 4.6.1.1] in the plasma membrane. An increase in cyclic AMP prior to receptor bridging suppressed phospholipid methylation in the plasma membrane, Ca2+ uptake, and subsequent histamine release. On the other hand, inhibition of phospholipid methylation by (S)-isobutyryl-3-deazaadenosine resulted in the suppression of cyclic AMP synthesis in the plasma membrane. These findings suggest that the activation of phospholipid methylation and the activation of adenylate cyclase are e, and subsequent histamine release. On the other hand, inhibition of phospholipid methylation by (S)-isobutyryl-3-deazadenosine resulted in the suppression of cyclic AMP synthesis in the plasma membrane. These findings suggest that the activation of phospholipid methylation and the activation of adenylate cyclase are e, and subsequent histamine release. On the other hand, inhibition of phospholipid methylation by (S)-isobutyryl-3-deazadenosine resulted in the suppression of cyclic AMP synthesis in the plasma membrane. These findings suggest that the activation of phospholipid methylation and the activation of adenylate cyclase are mutually regulated. PMID- 6273890 TI - Base insertion and deletion mutations induced in an Escherichia coli plasmid by benzo[a]pyrene-7,8-dihydrodiol-9,10-epoxide. AB - Mutations induced by (+/-)trans-benzo-[a]pyrene-7,8-dihydrodiol-9,10-epoxide (BaP diol epoxide-1) were selected by using a recombinant plasmid vector system designed for the study of transcription termination signals. The plasmid contains a transcription terminator positioned between a promotor signal and the Escherichia coli galactokinase structural gene (galK). By selections for the expression of galK (i.e., galK- to galK+), mutations are obtained in the terminator region that allow transcription from the promotor to read the galK gene. These mutations were characterized by direct DNA sequence of the terminator region. The DNA sequence changes caused by BaP-diol epoxide-1 were demonstrated for three different mutants. Two were found to be single-base-pair insertions of T.A into a cluster of consecutive T.A base pairs and the other change was a single-base-pair deletion of G.C from a cluster of consecutive G.C base pairs. PMID- 6273891 TI - Hemin inhibits ATP-dependent ubiquitin-dependent proteolysis: role of hemin in regulating ubiquitin conjugate degradation. AB - Hemin has been shown to inhibit specifically the energy-dependent degradation of normal and abnormal proteins in reticulocytes [Etlinger, J. D. & Goldberg, A. L. (1980) J. Biol. Chem. 255, 4563-4568]. The present work demonstrates that the action of hemin involves the multi-enzyme ATP-dependent ubiquitin-dependent proteolytic system exclusively. At a concentration of approximately 25 microM, hemin produces 50% inhibition of the degradation of 125I-labeled bovine serum albumin by this pathway. Hemin has no effect on the basal rate of proteolysis in the absence of either ATP or ubiquitin. At a concentration of hemin that gives complete inhibition of proteolysis, ATP-dependent formation of ubiquitin conjugates continues at about 50% of the control rate but the degradation of these ubiquitin conjugates is completely blocked. Inhibition of overall proteolysis and conjugate degradation are sensitive to hemin concentration to exactly the same extent. Hemin inhibition of conjugate breakdown results in the accumulation of higher molecular weight conjugates that are lost when hemin is removed by dilution. A model is proposed in which hemin acts as a negative allosteric effector in the initial step of a sequential degradative path by which intact ubiquitin conjugates are first cleaved to ubiquitin-associated fragments. PMID- 6273892 TI - Amino acid sequences of bacterial cytochromes c' and c-556. AB - The cytochrome c' are electron transport proteins widely distributed in photosynthetic and aerobic bacteria. We report the amino acid sequences of the proteins from 12 different bacterial species, and we show by sequences that the cytochromes c-556 from 2 different bacteria are structurally related to the cytochromes c'. Unlike the mitochondrial cytochromes c, the heme binding site in the cytochromes c' and c-556 is near the COOH terminus. The cytochromes c-556 probably have a methionine sixth heme ligand located near the NH2 terminus, whereas the cytochromes c' may be pentacoordinate. Quantitative comparison of cytochrome c' and c-556 sequences indicates a relatively low 28% average identity. PMID- 6273893 TI - Interactions between photoexcited rhodopsin and GTP-binding protein: kinetic and stoichiometric analyses from light-scattering changes. AB - In rod outer segments, photoexcited rhodopsin (R*) activates a cyclic GMP phosphodiesterase through a sequence of reactions involving a GTP-binding protein. By measuring light-scattering changes above 700 nm, we have studied the kinetics and stoichiometry of the association of R* with this protein and of the dissociation of the complex upon GDP/GTP exchange. Two light-scattering signals were obtained upon photoexcitation of rhodopsin in bovine rod outer segment membranes as well as in a reconstituted system consisting of purified GTP-binding protein and washed disc membranes; both signals depended specifically on the presence of GTP-binding protein. A "binding signal" that was observed in the absence of gTP as an increase in turbidity became saturated when a number of rhodopsin molecules equal to the number of GTP-binding protein molecules present (congruent to 10% in rod outer segments) has been bleached, suggesting that the protein binds to R* in a 1:1 complex. A "dissociation signal" of opposite sign, observed in presence of GTP at greater than or equal to 1 microM, is half maximal at 0.04% bleaching and saturated at 0.5% bleaching; it is interpreted as reflecting the dissociation of GTP-binding protein-R* complexes after GDP/GTP exchange on the GTP-binding protein, one R* being able to interact sequentially with about 100 GTP-binding protein molecules. The early time course of the binding signal is faster than that of the dissociation signal, and both signals take place in the 100-msec range at 20 degrees C. PMID- 6273894 TI - Clonal analysis of early and late stages of erythroleukemia induced by molecular clones of integrated spleen focus-forming virus. AB - The integrated proviral DNA of the polycythemia-inducing isolate of Friend spleen focus-forming virus (SFFVp) has been identified in rat cell clones nonproductively infected with this replication-defective erythroleukemia virus and cloned in phage lambda vectors. These lambda SFFVp recombinants, lambda SFFVp502 and lambda SFFVp542, contain endonuclease EcoRI inserts of size 7.4 and 8.2 kilobases, respectively, and include full copies of the SFFVp genome, along with host flanking sequences. Infectivity of the cloned SFFVp genomes was tested by a two-step DNA transfer procedure involving transfection of the cloned DNA into 3T3 mouse fibroblasts or cotransfer of the cloned DNA into thymidine kinase deficient 3T3 cells together with the cloned thymidine kinase gene of herpes simplex virus, followed by rescue of the transferred DNA by superinfection with a helper virus. Inoculation of the rescued virus into adult mice resulted in the appearance of spleen foci, rapid splenomegaly, and polycythemia. Early after infection, spleen cell populations contained large numbers of cells capable of forming small erythroid colonies in vitro (CFU-E) in the absence of erythropoietin. Late after infection, these mice contained cells capable of forming macroscopic colonies (CFU-FV) in vitro. These data indicate that molecular clones of SFFVp, in conjunction with a helper virus, induce the appearance of hemopoietic colony-forming cells characteristic of both the early and late stages of Friend leukemia. PMID- 6273895 TI - Nuclear DNA from uninfected or potato spindle tuber viroid-infected tomato plants contains no detectable sequences complementary to cloned double-stranded viroid cDNA. AB - High molecular weight tomato nuclear DNA was isolated from uninfected and potato spindle tuber viroid (PSTV)-infected tomato leaves. Restriction digests were fractionated on agarose gels, denatured and transferred to diazobenzyloxymethylpaper, and hybridized to 32P-labeled cloned double-stranded PSTV cDNA. No hybridization to DNA from either uninfected or infected tissue could be detected under conditions that permitted detection of cloned double stranded PSTV cDNA at a concentration equivalent to one-fifth copy of PSTV related DNA per haploid tomato genome. Hybridization of tomato DNA to 32P-labeled cloned soybean 18S and 28S ribosomal DNA sequences showed that the restricted nuclear DNA was suitable for hybridization to probes containing homologous sequences. Our results indicate that neither PSTV nor its complementary strand is transcribed from nuclear DNA but do not rule out the possibility of sequence homology between host DNA and a small portion of PSTV or its complement. PMID- 6273896 TI - Renin, angiotensins, and angiotensin-converting enzyme in neuroblastoma cells: evidence for intracellular formation of angiotensins. AB - The mechanism of formation of various peptide hormones in neuronal cells in the brain is not clear. The question of whether brain angiotensin II is formed by an extracellular mechanism as in the peripheral system or by an intracellular mechanism can be answered by using cloned cells in culture. We have screened several neuroblastoma cell lines of rat and mouse origin and found at least three cell lines that contain renin (EC 3.4.99.19), angiotensin-converting enzyme (dipeptidyl carboxypeptidase; peptidyldipeptide hydrolase, EC 3.4.15.1), and angiotensins I and II. This finding was interpreted to indicate that in these cells angiotensin formation takes place by an intracellular mechanism, in contrast to the extracellular mechanism well known to occur in plasma. This study also demonstrates the existence of viable and cloned cell lines that produce renin. PMID- 6273897 TI - Quantitation and characterization of a species-specific and embryo stage dependent 55-kilodalton phosphoprotein also present in cells transformed by simian virus 40. AB - A 55-kilodalton (kDal) protein was detected recently in primary cultures of day 12 mouse embryos by immunoprecipitation with serum from simian virus 40 (SV40) tumor-bearing hamsters (T serum), Preliminary evidence suggested that this protein was similar to a cellular 55-kDal protein induced after SV40 transformation of mouse cells. We now show that specific approximately 55-kDal [35S]methionine-labeled proteins precipitate from primary cultures of midgestation mouse, rat, and hamster embryos on addition of T serum or monoclonal antiserum prepared against the SV40-induced mouse 55-kDal proteins. The two dimensional maps of the [35S]methionine-labeled tryptic peptides of the mouse, hamster, and rat embryo proteins are similar to the maps of the corresponding proteins from SV40-transformed cells. Primary cells from midgestation mouse, hamster, or rat embryos contain one-third to one-half as much 55-kDal protein as a SV40-transformed mouse fibroblast cell and nearly the same amount as F9 mouse embryonal carcinoma cells. The amount of 55-kDal protein is greatly reduced on replating the mouse, rat, or hamster embryo primary cells. The amount of this protein in mouse embryos is dependent on the stage of the embryo. The embryo proteins are phosphoproteins. PMID- 6273899 TI - Rotational diffusion of epidermal growth factor complexed to cell surface receptors reflects rapid microaggregation and endocytosis of occupied receptors. AB - The rotational diffusion of epidermal growth factor (EGF)--receptor complexes on living human epidermoid carcinoma cells (A-431) has been measured by phosphorescence emission and anisotropy in the mu s time domain. A biologically active phosphorescent conjugate of EGF, erythrosin-EGF, was applied to living cells. The hormone--receptor complexes were mobile with rotational correlation times in the range 25--90 mu s when labeled and measured at 4 degrees C. Prolonged incubation and exposure to higher temperatures (23 and 37 degrees C) resulted in longer times up to 350 mu s, indicative of the progressive formation of microclusters, estimated to contain 10-50 receptors. Upon internalization of the hormone--receptor complexes, visible patches were observed by fluorescence microscopy, and the rotational correlation times were shorter, indicating a decrease in size of the dynamic unit. The sign of the rotational relaxation also varied with localization and processing of the hormones. The rate of lateral diffusion of EGF--receptor complexes, measured under similar conditions by fluorescence photobleaching recovery, increased with temperature in contrast to the rotational motion. PMID- 6273898 TI - Morphologic study of the internalization of a lysosomal enzyme by the mannose 6 phosphate receptor in cultured Chinese hamster ovary cells. AB - The binding and internalization of a model lysosomal enzyme, beta-galactosidase, was visualized by use of rabbit anti-beta-galactosidase and goat anti-rabbit IgG; the second antibody was labeled with rhodamine or fluorescein (for detection by fluorescence) or with horseradish peroxidase (for electron microscopy). Chinese hamster ovary cells were incubated with beta-galactosidase at 4 degrees C, and then were washed and sequentially incubated in the cold with the two antibodies. The beta-galactosidase was found primarily in coated pits. The binding of the enzyme was completely inhibited by 5 mM mannose 6-phosphate. After the reaction with enzyme and antibodies, the cells were warmed to 37 degrees C; within 1 minute, the beta-galactosidase--antibody complex had begun to move to uncoated vesicles (receptosomes). After 8 min, the beta-galactosidase--antibody complex was seen in receptosomes near tubular elements in the Golgi/GERL area, within such tubular elements and at times, in vesicular elements that may correspond to coated structures of the GERL system. After 15 min, the enzyme--antibody complex was found in lysosomes near the Golgi/GERL are and a half-hour later it was in lysosomes distributed throughout the cytoplasm. Double-label experiments using beta-galactosidase and gold/alpha 2-macroglobulin showed the presence of the two ligands in the same coated pits and receptosomes. Thus, the pathway for internalization of beta-galactosidase via the mannose 6-phosphate receptor is similar to the pathway established for other ligands such as low density lipoprotein and alpha 2-macroglobulin. PMID- 6273901 TI - Developmental onset of mixed-function oxidase activity in preimplantation mouse embryos. AB - Two-cell embryos, obtained from the C57BL/6N and DBA/2N strains, were cultured in media that supported in vitro differentiation and that contained [3H]benzo[a]pyrene. High-pressure liquid chromatography of the activated intermediates formed during in vitro early embryonic development indicated that the onset of polynuclear aromatic hydrocarbon activation coincided with blastocyst formation. Comparison of individual oxygenated intermediates metabolically formed from embryos genetically "responsive" or "nonresponsive" to aromatic hydrocarbons revealed significant quantitative differences in the production of dihydrodiol, quinone, and phenolic derivatives. In addition to exhibiting basal mixed-function oxidase activity, blastocysts were also responsive to enzymatic induction when exposed to 2,-3,7,8-tetrachlorodibenzo-p dioxin. The presence of operative metabolite-detoxifying pathways was also assayed. Enzymatic treatment of water-soluble metabolites with beta-glucuronidase or arylsulfatase revealed that neither glucuronic acid conjugates nor sulfate ester derivatives were present. These data, therefore, provide direct evidence that late preimplantation mouse embryos (day 3 1/2 of gestation) are similar to later developmental stages in having the enzymatic capability for xenobiotic activation and enzyme induction but are dissimilar with respect to their detoxification mechanism(s). Moreover, the ability of preimplantation embryos to activate directly polynuclear aromatic hydrocarbon to bioreactive intermediates may be of importance in assessing the ontological susceptibility of the developing embryo to carcinogenic or teratogenic chemicals. PMID- 6273900 TI - Association of microtubules and intermediate filaments in normal fibroblasts and its disruption upon transformation by a temperature-sensitive mutant of Rous sarcoma virus. AB - By double indirect immunofluorescence, using primary rabbit antibodies to tubulin and guinea pig antibodies to vimentin, we have simultaneously labeled microtubules and intermediate filaments in several types of cultured normal fibroblasts. With well-spread interphase cells there was an extensive but not complete correspondence of the labeling patterns for the two filamentous structures out to the cell periphery. This correspondence existed both at a gross level, where parallel but not coincident arrays of thickly labeled strands of the two types of filaments were observed, and at a fine level, where thinly labeled strands of the two were superimposed. The results suggest that there may be some type(s) of molecular linkages between microtubules and vimentin intermediate filaments that is under metabolic control. With NRK fibroblasts infected with a temperature-sensitive mutant (LA23) of Rous sarcoma virus, cells grown at the nonpermissive temperature (39 degrees C) showed the correspondence of the distributions of the microtubules and intermediate filaments characteristic of the normal phenotype but within 1 hr after a shift to the permissive temperature (33 degrees C) there was an extensive retraction of the intermediate filaments around the cell nucleus whereas the microtubules remained dispersed into the cell periphery. These results suggest that one of the functions carried out by p60src, the protein kinase responsible for transformation by Rous sarcoma virus, may be to modify the component(s) involved in the putative linkages between microtubules and intermediate filaments in the normal cells. PMID- 6273902 TI - Regional assignment of genes for human alpha-globin and phosphoglycollate phosphatase to the short arm of chromosome 16. AB - The human alpha-globin and phosphoglycollate phosphatase (EC 3.1.3.18) genes have been regionally localized to the short arm of human chromosome 16 (HC16). This was accomplished by fusing mouse fibroblasts (A9) to human fibroblasts that contain a reciprocal translocation between the long arms of chromosomes 16 and 11. The murine A9 cells are deficient in adenine phosphoribosyltransferase (APRT), an enzyme present on the long arm of HC16 (HC16q). Hybrid cells were grown in selection culture medium that required the cells to retain human APRT. Therefore, the hybrids exhibited stable retention of the entire HC16 or the rearranged chromosome containing HC16q. We isolated five independent primary and secondary hybrid cell lines which retained either HC16 or HC16q at a high frequency. The presence of human alpha-globin genes in the various clones was established directly by DNA extraction and hybridization to a cDNA probe for human alpha-globin genes. Autoradiographs showed that hybrid cells containing the long arm, but not the short arm, of HC16 showed only the background mouse bands. Hybrid cells that retained the entire HC16 demonstrated the band(s) containing the human alpha-globin genes. Hybrid cells that contained HC16 with its alpha globin genes were then placed in culture medium that contained diaminopurine, which is lethal for cells containing APRT. These counter-selected hybrid cells had lost HC16 and also lost the human alpha-globin genes as determined by blot hybridization. The presence of alpha-globin gene sequences in the hybrid clones was concordant with HC16 only and not with any other human chromosome. These results confirm the assignment of alpha-globin genes to HC16 and localize the genes to the short arm. We also assign the locus for phosphoglycollate to the short arm of HC16. PMID- 6273903 TI - Cloning of chromosomal genes in Streptococcus pneumoniae. AB - A system for molecular cloning in Streptococcus pneumoniae was developed. The multicopy plasmids pMV158 (5.4 kilobases) and pLS1 (4.3 kilobases), which confer tetracycline resistance, were used as vectors to clone chromosomal genes of S. pneumoniae in host cells of this species. A 3.3-kilobase restriction fragment containing the malM gene, which codes for amylomaltase, was cloned in a deletion mutant lacking chromosomal homology with the fragment. The recombinant plasmid pLS70, could transform over 50% of a recipient population to maltose utilization. Amylomaltase constituted up to 10% of the protein of cells containing pLS70. A derivative with a deletion, pLS69, appeared to gain a selective advantage by producing less enzyme. A 10-kilobase restriction fragment containing the sul-d gene for sulfonamide resistance was cloned in the presence of the homologous chromosomal gene. De novo establishment of a recombinant plasmid was just as frequent as transformation in an endogenous plasmid. Despite the processing of DNA during uptake in the transformation of S. pneumoniae, recombinant plasmids can be introduced. Models for the reconstruction of recombinant DNA in cells of S. pneumoniae and Bacillus subtilis are considered and compared. PMID- 6273904 TI - Integration, transcription, and control of a Drosophila heat shock gene in mouse cells. AB - Mouse L cells were transformed with a cloned 3.6-kilobase (kb) segment of Drosophila melanogaster DNA carrying the 2.25-kb transcribed sequence for the Drosophila 70,000-dalton heat shock protein (hsp70) and 1.1 kb and 0.2 kb of 5' and 3' flanking DNA, respectively. Heat shock of one of three such transformed cell lines containing multiple copies of the intact Drosophila segment induced the abundant accumulation of transcripts of the Drosophila gene, with correct or nearly correct 5' and 3' termini. This provides evidence, in accord with earlier indications, that diverse eukaryotes, including vertebrates, have heat shock systems similar to that studied extensively in Drosophila. Our results suggest that the signals for heat shock transcription and the chromosomal sites with which they interact have been highly conserved in evolution and that the regulatory sequences controlling transcription of the gene for hsp70 lie within the 3.6-kb Drosophila segment. PMID- 6273905 TI - Site-specific inversion sequence of the herpes simplex virus genome: domain and structural features. AB - The genome of herpes simplex virus-1 consists of two covalently linked components, L and S, that invert relative to each other. The L and S components consist of unique DNA sequences bracketed by inverted repeats. The inverted repeats of the L component are designated ab and b' a' and those of the S component are designated a' c' and ca. The number of a sequences at the termini and at the L-S component junction varies from one to several copies. Insertion into the middle of the L component of a DNA fragment consisting of 156 base pairs (bp) of the b sequence, an entire a sequence of 501 bp, and 618 bp of the c sequence created a new site through which additional inversions in the genome occurred. Comparison of the nucleotide sequences of DNA fragments containing one and two a sequences defined the domain of the a sequence. The single a sequence consists of two 20-bp direct repeats (designated as DR1) bracketing a region that contains 19 tandem direct repeats of a 12-bp sequence (DR2) adjacent to three direct repeats of a 37-bp sequence (DR4), in addition to short stretches of unique sequences. The fragment with two tandem a sequences contained three copies of DR1-i.e., the intervening DR1 was shared by the two a sequences. Furthermore, one a sequence contained 22 copies of DR2 and two copies of DR4 whereas the second a sequence contained 19 copies of DR2 and two copies of DR4. These observations suggest that (i) amplification of the number of terminal and internal a sequences is the consequence of intramolecular or intermolecular recombination through DR1, (ii) the number of copies of DR2 and DR4 within the a sequence is not fixed and may vary as a consequence of unequal crossing over or slippage, and (iii) inversion results from intramolecular recombination between terminal and inverted a sequences. PMID- 6273907 TI - A common mutant EcoRI restriction endonuclease site in the 5' flanking portion of the human alpha-globin gene. AB - A mutant EcoRI endonuclease restriction site has been identified in 3 of 37 Black subjects and in 2 sibs of one of these persons. This mutation was not encountered in 13 Whites. It is located approximately 6 kilobases "inside" the normal site in the 5' flanking sequence of the alpha-globin chain complex. The shortened alpha globin gene-bearing segment produced in the EcoRI digest produces a restriction map similar to that observed for the common alpha-globin gene deletion observed in the Black population. However, the restriction map with BamHI is normal, confirming that all four alpha loci are present. PMID- 6273906 TI - Superoxide dismutase in Drosophila melanogaster: biochemical and structural characterization of allozyme variants. AB - Superoxide dismutase (SOD; superoxide:superoxide oxidoreductase, EC 1.15.1.1) is known to be polymorphic in many organisms; in Drosophila, the degree of polymorphism has a wide range of variation from locality to locality within a given species. We have thoroughly purified from D. melanogaster the two common electromorphs, SODS and SODF. These differ in properties such as isoelectric point, specific activity, rate constant, thermostability, and amino acid composition. The specific activity is three times greater in SODS than in SODF, but the latter is more thermostable. In strains from California, SODS differs from SODF by at least one amino acid substitution: lysine in SODS is replaced by either aspartic acid or asparagine in SODF. This difference is consistent with the electrophoretic mobility and isoelectric points of the two electromorphs. In strains from Africa, SODS and SODF differ by two amino acid substitutions (histidine and proline in SODS vs. serine and either glutamic acid or glutamine in SODF) in addition to the one distinguishing the California strains. Thus the SODF electromorphs from California and from Tunisia, in spite of their identical electrophoretic mobility, differ by at least two amino acid substitutions. PMID- 6273908 TI - Allelic forms of rat kappa chain genes: evidence for strong selection at the level of nucleotide sequence. AB - The genes that code for two allotypic forms of the rat kappa light chain constant region (C(kappa)) have been cloned and the nucleotide sequence of 1172 base pairs of coding and flanking sequence has been determined for both alleles. These sequences have been compared to each other and to the corresponding sequences found in the mouse and human. Comparison of the LEW allele with mouse C(kappa) reveals two surprising features: (i) There is an unusually large number of amino acid substitutions (21) relative to the total number of nucleotide changes (37) in the coding region. Comparision among several other mammalian genes reveals a larger proportion of "silent" changes. (ii) The rate of accumulation of base substitutions is the same within the coding region as it is in some 870 base pairs of noncoding sequence (including 3' untranslated, 3' flanking, and 5' intervening sequences). Comparison of the two allelic forms of rat C(kappa) shows the same unusual features in more extreme form. (i) Twelve base substitutions in the coding region determine 11 amino acid differences-only one "silent" change exists. (ii) There are 12 base substitutions in the 318 base pairs of coding sequence (3.7% difference) and only 9 in the remaining 854 base pairs of noncoding DNA (1.1%), a highly significant difference. This degree of conservation of noncoding sequences and of "silent" sites within the coding region is unique among the mammalian genes studied thus far. These patterns suggest that there has been strong selection for conservation of nucleotide sequences, both inside and outside the coding region, independent of the selection required to maintain the function and characteristic structure of the immunoglobulin domain itself. The functions of the nucleotide sequences that account for this selective pressure are unclear at the present time. PMID- 6273909 TI - Inversions between ribosomal RNA genes of Escherichia coli. AB - It might be anticipated that the presence of redundant but oppositely oriented sequences in a chromosome could allow inversion of the intervening material through homologous recombination. For example, the ribosomal RNA gene rrnD of Escherichia coli has the opposite orientation fro rrnB and rrnE and is separated from these genes by roughly 20% of the chromosome. Starting with a derivative of Cavalli Hfr, we have constructed mutants that have an inversion of the segment between rrnD and either rrnB or rrnE. These mutants are generally quite viable but do exhibit a slight reduction in growth rate relative to the parental strain. A major line of laboratory E. coli, W3110 and its derivatives, also has an inversion between rrnD and rrnE, probably created directly by a recombinational event between these highly homologous genes. PMID- 6273912 TI - A plausible two-state model for cytochrome c oxidase. AB - The catalytic properties of pulsed and resting cytochrome c oxidase (ferrocytochrome c: oxygen oxidoreductase, EC 1.9.3.1), expressed in terms of a minimal kinetic scheme and simulated by numerical computations, were successfully described. A two-state model, in which the relative amounts of the enzyme present in each conformation are regulated by the rates of electron flux and O2 binding on one side and the interconversion rates on the other, accounts for the activation of cytochrome c oxidase during turnover. PMID- 6273910 TI - Two alpha heavy chain disease proteins with different genomic deletions demonstrate that nonexpressed alpha heavy chain genes contain methylated bases. AB - Two independently arising alpha heavy chain mutants have been found to synthesize heavy chains with CH1 deletions of approximately equal extent. Both were isolated from heavy chain-producing variants of the mouse myeloma W3129 and demonstrate that it is possible to arrive at the heavy chain disease phenotype by the pathway H + L leads to H leads to delta H. Analysis of genomic DNA by digestion with restriction endonucleases followed by molecular hybridization showed that one mutant (delta 37) had a deletion of approximately 0.2 kilobase and the second mutant (delta 15) had a deletion of approximately 0.5 kilobase. Mouse myeloma cells contain several alpha chain alleles but only one is expressed; the presence of the deletion in delta 37 and delta 15 made it possible to identify the restriction fragments from the expressed allele. Analysis of the fragments produced after cleavage with an isoschizomeric pair of restriction enzymes, Msp I and Hpa II, indicated that, in the W3129 cell line and its variants, the unexpressed alpha alleles contain methylated bases. The influence of methylation on gene expression remains to be elucidated. PMID- 6273911 TI - Developmental hierarchy of immunoglobulin gene rearrangements in human leukemic pre-B-cells. AB - We have used a special class of human acute lymphocytic leukemias, the common "non-T/non-B" cell type, to define a hierarchy of genetic rearrangements that occur during the earliest stages of B-cell maturation. This has allowed us to identify intermediate cells predicted by a hierarchial model in which immunoglobulin heavy chain variable region gene formation precedes that of light chain and in which kappa light chain gene formation precedes that of lambda. The model emphasizes the flexible nature of immunoglobulin gene recombination that not infrequently produces aberrant or null genes that are phenotypically excluded from expression. Remaining alleles or isotypic genes can then be utilized as "spares" undergoing recombination until a valid gene is formed. Significantly, the excluded allele or isotype is frequently deleted from the genome. In addition to defining a pathway of genetic maturation, this analysis provides a powerful means to further classify cases of non-T/non-B-cell acute lymphocytic leukemia, most of which seem to reside at early stages along the B-cell pathway of differentiation. PMID- 6273913 TI - Suppression of malignancy in human cancer cells: issues and challenges. AB - Analysis of the many, sometimes seemingly contradictory, reports on the partial suppression of malignancy in highly unstable rodent intraspecies and rodent- human hybrid cells emphasizes the limitations of this approach to the analysis of the basic nature of malignancy, especially in naturally occurring human cancers. During the past 5 years, Stanbridge and then Klinger reported complete suppression, not elimination, of malignancy [defined as capacity to produce progressively growing tumors in athymic (nude) mice] in stable hybrids of different human cancer cells with normal human fibroblasts or with differentiating epithelial keratinocytes and, importantly, also in stable hybrids of two parental cancers of different somatic cell origin. The nontumorigenic human hybrid cells are not rejected by some nonthymic immune mechanism of nude mice and survive in vascularized foci; the initial multiplication of these cells is stopped by some unknown proliferation controlling substance(s) to which their malignant parent(s) do not respond. The heritable properties of infinite multiplication in vitro, loss of contact inhibition, etc. remained in the nontumorigenic hybrids but, remarkably, the in vitro production of alpha human choriogonadotropin by HeLa cells was suppressed along with tumorigenicity and reappeared in the tumorigenic revertants. If it is assumed that human cancers of different somatic cell origin are caused by a loss of different specific regulatory genes, as the most recent data reviewed here suggest, the challenge is to determine in molecular terms what those missing genes are, how they function, and whether it may be possible to restore to the cancer cells what they have lost. PMID- 6273914 TI - Plasma cell dyscrasia and peripheral neuropathy: identification of the myelin antigens that react with human paraproteins. AB - In some cases of polyneuropathy and plasma cell dyscrasia, the monclonal antibodies react with human peripheral nerve myelin. To identify the myelin antigens involved, we separated the proteins of human central and peripheral nerve myelin by polyacrylamide gel electrophoresis, transferred the proteins onto nitrocellulose sheets, and used an immunoenzymatic technique to detect the reactive antigens. Serum IgM but not IgG from three patients with neuropathy and complement-fixing anti-human myelin IgM paraproteins immunostained a protein of approximately 100,000 daltons in human peripheral nerve myelin and a protein or closely migrating proteins of similar size in human central nervous system myelin. In a fourth patient, both IgM and IgG immunostained the antigen. Immunostaining was specific for the paraprotein light chain type, and absorption of the patients' sera with human peripheral nerve myelin eliminated the reaction with the central nervous system proteins. No reaction was seen with rabbit peripheral nerve myelin or with membranes prepared from human myotubes, human T cells, or human fibroblasts. Control sera from six patients with neuropathy and IgM paraproteins that did not react with myelin, from four patients with IgM paraproteins but no neuropathy, and from three normal subjects did not immunostain myelin. PMID- 6273915 TI - An Epstein--Barr virus early protein induces cell fusion. AB - Superinfection of Raji cells with Epstein--Barr virus (EBV) leads to syncytium formation. Studies using metabolic inhibitors and amino acid analogues suggest that the fusion-inducing factor belongs to the early group of virus-specified proteins. Induction of early EBV protein synthesis in Raji cells by using various chemicals also leads to syncytium formation, indicating that the fusion process is not caused by a virion membrane protein introduced into the cells upon infection. The relevance of these findings to the association of EBV with carcinoma of the nasopharynx is discussed. PMID- 6273916 TI - Detergent extraction of a presumptive gating component from the voltage-dependent sodium channel. AB - A physiologically characterized radiolabeled neurotoxin complex obtained from venom of the scorpion Leiurus quinquestriatus has been used to identify detergent solubilized presumptive sodium channel components in sucrose gradients. This toxin-binding component is found in extracts prepared from three sources of excitable membrane but appears to be absent from similar extracts prepared from nonexcitable membrane or from Torpedo californica membrane. Procedures that destroy the physiological activity of the Leiurus neurotoxin lead to a corresponding loss of toxin binding to the putative sodium channel component. The major component recognized by the Leiurus toxin sediments at 6.5 S. Scatchard analysis of quantitative binding experiments carried out in sucrose gradients shows approximately linear plots and indicates that the toxin recognizes a relatively small number of sites with a dissociation constant near 10 nM. Once formed, the channel element--toxin complex is quite stable. Experiments show diphasic dissociation kinetics with half-times near 70 hr and greater than 200 hr. PMID- 6273917 TI - Electrical stimulation of sympathetic nerves increases the concentration of cyclic AMP in rat pineal gland. AB - Electrical stimulation of the superior cervical ganglia causes a rapid increase in the concentration of cyclic AMP in the pineal gland of rats. This effect is dependent upon the frequency, voltage, and duration of the stimulus and is markedly potentiated by pretreating the animals with desmethylimipramine. The increase in cyclic AMP is blocked by prior treatment of the rats with reserpine, bretylium, or propanolol but not with phentolamine. These results provide direct evidence that electrical stimulation of sympathetic nerves increases cyclic AMP in a target organ through the release of norepinephrine from presynaptic terminals acting on postsynaptic beta-adrenergic receptors. PMID- 6273918 TI - Diazepam and (--)-pentobarbital: fluctuation analysis reveals different mechanisms for potentiation of gamma-aminobutyric acid responses in cultured central neurons. AB - Diazepam and (--)-pentobarbital each potentiate the increase in chloride ion conductance produced by gamma-aminobutyric acid (GABA) i voltage-clamped mouse spinal neurons grown in culture. Fluctuation analysis was used to compare the properties of elementary ion-channel events underlying the chloride conductance produced by GABA alone and during potentiation by the two drugs. Neither drug altered the conductance of an open ion channel, but both drugs affected the kinetics of channel activity. Diazepam increased the frequency of channel openings and either did not affect or slightly increased the average open-channel lifetime, whereas (--)-pentobarbital decreased the frequency of channel openings and increased average open-channel lifetime. These changes in the kinetics of GABA-activated ion channels can quantitatively account for the potentiation of GABA responses observed with the drugs. Thus, the drugs each increase the response to GABA but do not act on channel kinetics in the same manner. PMID- 6273920 TI - Are the presynaptic membrane particles the calcium channels? AB - The number of large intramembrane particles associated with sites of synaptic vesicle release at the squid giant synapse was determined and compared to the average maximal presynaptic calcium current in order to derive an estimate of the conductance each particle would have if it were a calcium channel. This value, 0.21 pS, compares favorably with conductances of calcium channels in other preparations, substantiating the idea that the large intramembrane particles, which are concentrated at "active zones," represent calcium channels. PMID- 6273921 TI - Estimate of incidence and direct economic losses due to bluetongue in Mississippi cattle during 1979. PMID- 6273919 TI - Mutational alteration of membrane phospholipid composition and voltage-sensitive ion channel function in paramecium. AB - A behavioral mutant of Paramecium tetraurelia (baA) has been isolated that has an abnormal response when placed in solutions containing Ba2+. This mutant is shown here to have a dramatic alteration of the sphingolipid and phosphonolipid composition of its ciliary membrane. This biochemical defect is present in independently isolated alleles at baA locus and segregates in crosses with the behavioral phenotype. Electrophysiologically, the mutation reduces significantly conductance of both voltage-sensitive Ca2+ channels and voltage-sensitive K+ channels. When the mutant is grown in sterol-supplemented medium, its behavior, electrophysiological properties, and lipid composition are hardly distinguishable from wild type grown under similar conditions. This mutant then, provides strong evidence that membrane lipids significantly influence the function of the membrane molecules responsible for the generation of action potentials. PMID- 6273922 TI - Blood autograft in sheep for isolation of bluetongue virus from latently infected cattle. PMID- 6273923 TI - Bovine viral diarrhea virus (BVDV) serotiters stimulated in cattle in isolation and under field conditions by inactivated BVDV vaccine. PMID- 6273924 TI - Aflatoxin in human and animal health. PMID- 6273925 TI - Marihuana and alcohol: perinatal effects on development of male reproductive functions in mice. AB - Marihuana and alcohol have been reported to exert a wide range of effects on reproductive functions in man and laboratory animals. Decreased testosterone levels, suppression of pituitary gonadotropin release, impaired fertility and sexual dysfunction have been observed in adult males exposed to either of these substances. The present studies examined the effects of perinatal exposure to ETOH and THC alone or in combination on the development or reproductive functions in mice. Testes weights and plasma testosterone levels were reduced in adult males perinatally exposed to these substances, alone or in combination. Seminal vesicle weights were reduced only in the THC-exposed mice, while kidney weights were decreased by either ETOH or THC exposure. In ETOH plus THC-exposed animals, plasma LH was reduced while FSH levels were increased. In response to castration, gonadotropin levels were elevated in mice perinatally exposed to THC. In vitro responsiveness to gonadotropin stimulation, as indicated by testosterone production by decapsulated testes, was significantly inhibited in tissue obtained from ETOH-exposed males. Although no simple interactions were observed, it is evident that THC and ETOH, either alone or in combination, can affect the development of male reproductive functions in mice. PMID- 6273926 TI - Effect of ethanol on spontaneous and stimulated growth hormone secretion. AB - Growth hormone (GH) regulates not only somatic growth but also carbohydrate, protein and lipid metabolism. Altered secretory states resulting from alcohol use could have pathogenetic significance. The pattern of spontaneous GH secretion in man is reviewed and previously published human studies on ethanol and GH examined to develop a coherent formulation of the nature of ethanol effects on GH secretion. The weight of the evidence suggests that ethanol suppresses GH secretion. Studies using the rat indicates that the dose-response relationship is of a threshold nature, with doses of 3 g/kg or greater abolishing spontaneous secretion. The possible role of diminished GH secretion in the pathogenesis of the fetal alcohol syndrome is discussed. PMID- 6273927 TI - Phospholipid methylation: a possible mechanism of signal transduction across biomembranes. AB - The conversion of phosphatidylethanolamine (PE) to phosphatidylcholine (PC) is catalyzed by two methyltransferases with S-adenosylmethionine as the methyl donor. PC formed by transmethylation is further metabolized by phospholipase A2. The synthesis and degradation of methylated phospholipids are involved in regulating the number of the beta-adrenergic receptors and their coupling to adenylate cyclase in rat reticulocytes, HeLa cells, and rat astrocytoma cells. Methylation of the phospholipids in these cells is stimulated by binding of agonists to the beta-adrenergic receptors. Accumulation of phosphatidyl-N monomethylethanolamine causes an increase in membrane fluidity and enhances the coupling of the receptors to adenylate cyclase. Agents that inhibit phospholipid methylation decrease the number of receptors in intact HeLa cells, while increased phospholipid methylation unmasks cryptic receptors. Conversely, the degradation of methylated phospholipids appears to be closely associated with the desensitization of the beta-adrenergic receptors following prolonged stimulation with isoproterenol. Inhibition and stimulation of phospholipase A2 causes inhibition and stimulation of this desensitization process. PMID- 6273928 TI - Human mutation affecting hormone-sensitive adenylate cyclase. AB - Hormone-sensitive adenylate cyclase contains a recently discovered protein component that is required for stimulation of cyclic AMP synthesis by hormones and guanine nucleotides. We measured this protein in erythrocyte membranes of ten patients with pseudohypoparathyroidism (PHP), using assays of its biochemical activity and of its susceptibility to radiolabeling in the presence of 32P-NAD and cholera toxin. By both assays, the protein was reduced by 50% in erythrocytes of 4 PHP patients, as compared with normal and hypoparathyroid subjects. These 4 subjects, in contrast to the 6 PHP patients (5 in one family) whose erythrocytes contained apparently normal amounts of the cyclase component, exhibited the full spectrum of skeletal abnormalities found in PHP. We conclude that partial deficiency of the guanine nucleotide regulatory protein is a biochemical marker for a subset of PHP patients. If present in other tissues, this deficiency could explain the resistance of target organs in PHP to parathormone and other hormones that work via cyclic AMP. PMID- 6273929 TI - Epidermal growth factor-receptor-protein kinase interactions. AB - Membranes prepared from A-431 human epidermoid carcinoma cells retained the ability to bind 125I-labeled epidermal growth factor (EGF) in a specific manner. In the presence of [gamma-32]ATP and Mn2+ or Mg2+, this membrane preparation was capable of phosphorylating specific endogenous membrane proteins, as well as exogenously added histone. The binding of EGF to membranes in vitro resulted in a several-fold stimulation of the phosphorylation reaction. The phosphorylation reaction was not dependent on cyclic AMP or cyclic GMP. These findings suggest that one of the biochemical consequences of the binding of EGF to membranes is a rapid activation of a cyclic AMP-independent phosphorylating system. The activation of the membrane-associated protein kinase by EGF appears to be a reversible phenomenon. The membrane preparation could be solubilized by a number of non-ionic detergents with the retention of both 125I-labeled EGF binding activity and EGF-enhanced phosphorylation of specific membrane proteins. The solubilized membrane preparation was purified by affinity chromatography. The purified preparation retained both EGF-binding activity and EGF-enhanced phosphorylation activity. Analysis of the affinity-purified preparation by SDS gel electrophoresis indicated the presence of one major protein band of molecular weight 150,000 and several trace bands. The evidence suggests that the major 150,000 protein band is the receptor for EGF and is a substrate of the phosphorylation reaction. The co-purification of EGF-binding activity and EGF stimulated phosphorylation activity suggests an inherent close relationship. The EGF-stimulated phosphorylation reaction appears to be specific for tyrosine residues and in this regard resembles the src protein kinase. PMID- 6273930 TI - Definition of the transcription unit of the natural ovalbumin gene. PMID- 6273931 TI - The regulation of SV40 gene expression in nonpermissive cells. AB - The limitations to SV40 growth in nonpermissive cells are poorly understood. In differentiated mouse cells, early mRNA and T-antigens are synthesized, but no viral DNA replication has been detected. A plausible explanation for the limitation to viral DNA synthesis in these cells might be the inability of a mouse cell-specific DNA polymerase to interact with the SV40 T-antigen-viral DNA complex. In spite of this abortive viral-cell interaction, SV40 late viral transcripts can be detected in infected mouse cells. Both early and late transcripts can be detected in infected mouse cells. Both early and late SV40 transcriptional activities peak between 10 and 15 h post infection; by 24 h after infection SV40 RNA is almost undetectable. In spite of the detection of late SV40 mRNA in mouse cells, we have been unable to detect any structural proteins (VP1, VP2, or VP3) encoded by these transcripts. A more restrictive interaction occurs between SV40 and undifferentiated mouse teratocarcinoma cell lines. Using an in vitro technique, the viral transcriptional complex (VTC) assay, we were able to demonstrate viral transcriptional activity on both the early and the late strands of SV40 in F9 embryonal carcinoma cells. Nevertheless, no mature processed mRNAs or viral encoded polypeptides were detected in these cells. After differentiation of the F9 cells with retinoic acid, however, spliced early mRNAs, as well as the SV40 T-antigen, were present. PMID- 6273932 TI - Rh hemolytic disease of the newborn: its cause and prevention. PMID- 6273934 TI - Epidemiology of Rh hemolytic disease of the newborn, United States, 1960-1979. PMID- 6273933 TI - Current clinical concepts in hemolytic disease and blood group incompatibility. PMID- 6273935 TI - Histocompatibility antigens in pregnancy, abortions, infertility, preeclampsia, and trophoblast neoplasms. PMID- 6273936 TI - The human syncytiotrophoblast microvillous plasma membrane. PMID- 6273937 TI - A novel female influences delta 9-THC effects on plasma hormone levels in male mice. AB - Exposure to delta 9-tetrahydrocannabinol (THC) (50 mg/kg) alters the endocrine responsivity of male mice to female-related exteroceptive stimuli. Exposure to a novel female prevents or delays the THC-induced decrease in plasma testosterone (T) and luteinizing hormone (LH) levels. These hormonal alterations are apparently not due to the LH-releasing effects of female-related pheromonal or tactile cues, since administration of luteinizing hormone releasing factor (LRF) did not mimic the effects of a novel female on plasma T levels in THC-treated males. Exposure to a much lower dose of THC (0.5 mg/kg) did augment the LRF induced increases in plasma T levels suggesting a possible synergism between gonadotropins and THC on androgen production. The present findings suggest that THC-induced alterations in hormonal status may be influenced by complex social or environmental factors. PMID- 6273938 TI - Pharmacologic analysis of sodium valproate-induced suppression of secondary components of visual evoked potentials in albino rats. AB - Sodium Valproate (VPA) administered to rats in a dose of 10 or 200 mg/kg IP suppressed the slow negative wave (SNW) and photically-induced afterdischarge (SAD) of VEP (when they were present) within 15-30 min. The recovery of VEP amplitude began at 3 hr. This effect was antagonized by subconvulsive doses of convulsant benzodiazepine RO 5-3663 (2 mg/kg) and metrazol (15 mg/kg) but not by picrotoxin (2 mg/kg) and naloxone (10 mg/kg). The SNW suppression may be attributed to a disinhibitory action of a system located presynaptically on recurrent collaterals of the output neurons, or nerve terminals of inhibitory interneurons or both. Alternative conjecture suggests that VPA depolarizes the dendritic tree masking thereby somatic inhibition produced by recurrent circuits. PMID- 6273939 TI - Dopamine induced changes in L-fucose incorporation into proteins of rat hippocampus and corpus striatum during postnatal development. AB - In 60 day old (adult) male Wistar rats dopamine caused a dose-dependent increase of L-fucose incorporation into total proteins of both hippocampus and corpus striatum slices up to +47.8 +/- 6.0% (n=6) and +53.2 +/- 8.5% (n=20), respectively, when compared to corresponding controls. Under these conditions the dopamine concentration leading to a maximum stimulation of fucose incorporation was 5 X 10(-4) M in hippocampus and 1 X 10(-3) M in corpus striatum. In the latter tissue the range of dopamine concentrations causing a significant elevation in incorporation rates was larger than in hippocampal tissue. In the corpus striatum of 9 day old rats dopamine was ineffective, but by 30 days the transmitter stimulated fucose incorporation rate reached the maximum observed for any age studied. This developmental pattern seems to be related to the ontogenesis of dopamine receptor sites or dopamine sensitive adenylate cyclase formation in this brain structure. In the hippocampus the postnatal development of dopamine induced augmentation of glycoprotein synthesis showed a longer latency, but the maximum effect was also seen in 30 day old animals. These results support our assumption that at the end of the postnatal differentiation period the glycoprotein synthesis in brain tissue may be controlled (at least to some extent) by the state of dopaminergic receptors and/or of dopamine sensitive adenylate cyclase. PMID- 6273940 TI - Tetrahydrocannabinol potentiates reserpine-induced hypokinesia. AB - Delta-9-tetrahydrocannabinol (THC), a substance in marihuana, was found to produce a profound potentiation of reserpine-induced hypokinesia in rats as measured with a bar test. In these experiments, THC had no hypokinetic effect by itself but produced a more than 20-fold increase in the hypokinesia produced by reserpine. Reserpine-induced hypokinesia has been viewed as animal model of Parkinson's Disease. THC potentiation of reserpine-induced hypokinesia was observed to be both time- and dose-dependent (1 to 10 mg/kg THC). When administered by gavage to reserpine-pretreated subjects (7.5 mg/kg IP, 24 hours before), THC produced a potentiation of hypokinesia that developed fully within 1 hour, lasted at least 5 hours, and was absent by 12 hours after THC administration. This THC effect was slightly increased by physostigmine, a cholinesterase inhibitor, relatively unaffected by scopolamine, a muscarinic antagonist, and almost completely blocked by ethopropazine, an anticholinergic antiparkinson drug. The effect was completely unaffected by naloxone. Insofar as reserpine has been used with some clinical efficacy in hyperkinetic movement disorders such as Huntington's disease and tardive dyskinesia, it may be that potentiation of reserpine's hypokinetic effect by a drug such as THC could greatly increase the clinical value of reserpine or related drugs in the treatment of these disorders. PMID- 6273941 TI - Pancreatic beta-endorphin-like polypeptides. AB - Acid extracts of porcine pancreas acetone powders (but not of other tissues) have been shown to contain material which will react firstly in a radioimmunoassay for beta-endorphin, and secondly, compete with 3H-Naloxone and 3H-enkephalinamide in the radio-receptor assay. This material had a molecular weight of approximately 7,000 as judged by both P-10 and G-50 exclusion chromatography, even in the presence of 4 M urea. The G-50 partially purified and desalted fraction demonstrated a dose-dependent analgesia in mice and also influenced mouse behavior in vivo in a manner analogous to that produced by synthetic human beta endorphin. PMID- 6273942 TI - Inhibitory effects of delta 9-tetrahydrocannabinol on glycolytic substrates in the rat testis. AB - Experiments conducted with a rat testicular tissue indicated that delta 9 tetrahydrocannabinol (THC) produced a dose-dependent inhibition of glucose metabolism. Incubation of testicular tissue with different U-14C-fructose concentrations and 0.3 mM THC also significantly inhibited 14CO2 production; however, testicular utilization of 214C-pyruvate remained unaffected by THC. In other experiments, 0.3 mM THC significantly decreased tissue concentrations of dihydroxyacetone phosphate, fructose-1,6-diphosphate, and glucose-6-phosphate by 23, 31 and 29%, respectively. Uptake studies with 14C-2-deoxy-D-glucose showed a 43% reduction in uptake of this substrate by testicular tissue in the presence of 0.3 mM THC. These studies demonstrate that THC has an inhibitory effect on the utilization of energy substrates by the testis. Sites of THC action may be the membrane uptake step, the hexokinase step, and/or possible the phosphofructokinase step. It is proposed that this inhibition of testicular glycolysis could deprive the cells of their energy reserves and thereby may disrupt many gonadal functions. PMID- 6273943 TI - Effect of opiates on the caudate spindle in the cat. AB - The effect of opiate drugs on the caudate spindle (CS) in the cat was observed following both systemic and intracaudal administration. Systemic administration of morphine, pentazocine and pethidine inhibited the CS. The inhibitory effects of opiates were antagonized by lysergide and methysergide but not by naloxone, chlorpromazine, dehydrobenzperidol, amphetamine and atropine. Contrary to their systemic administration, the investigated opiates morphine, pentazocine, pethidine and Met5-Enkephaline enhanced the CS, when injected into the caudate nucleus close to the site of stimulation. Those facilitating effects of opiates on the CS were completely blocked by naloxone. The results suggest that serotoninergic mechanisms might be involved in the action of opiates on the neuronal activity of the caudate nucleus. PMID- 6273944 TI - Eritadenine: a new tool for investigation of the adenosine P site in plasma membranes of rat fat cells. AB - Eritadenine was investigated for its effects on adenylate cyclase activity of rat fat cell plasma membranes on cyclic AMP accumulation and lipolysis in isolated rat fat cells. In rat fat cell plasma membranes, a 50% inhibition of noradrenaline (100 mumol/l) stimulated adenylate cyclase was obtained at 11.6 mumol/l eritadenine or 9.0 mumol/l adenosine. NaF (3 mmol/l) stimulated adenylate cyclase was inhibited at concentrations of eritadenine (IC25 5.8 mumol/l) lower than those of adenosine (IC25 51.9 mumol/l). Eritadenine ethyl ester (100 mumol/l) was almost ineffective on adenylate cyclase. The inhibitory effect of eritadenine was resistant to adenosine deaminase. Isolated rat fat cells eritadenine (100 mumol/l) was completely ineffective to block noradrenaline stimulated cyclic AMP accumulation or lipolysis stimulated by theophylline or adenosine deaminase. It is suggested that eritadenine is an effector of the adenosine P site of fat cell plasma membranes. PMID- 6273945 TI - The human behavioral pharmacology of the common core heptapeptides. PMID- 6273946 TI - In vivo isolation of a pPJ3a-lambda cosmid mediated by the Tn2301 transposon. PMID- 6273947 TI - Physical mapping of the pea chloroplast DNA and localization of the ribosomal RNA genes. PMID- 6273948 TI - Genetic structure of the IncN plasmid N3. PMID- 6273949 TI - Transcription in yeast mitochondria: analysis of the 21 S rRNA region and its transcripts. PMID- 6273950 TI - Protein synthesis induced by infection with packaged lambda dv plasmid. PMID- 6273951 TI - A host-dependent hybrid plasmid suitable as a suicidal carrier for transposable elements. PMID- 6273952 TI - Mechanism of mitochondrial DNA replication in mouse L cells: localization of alkali-sensitive sites at the two origins of replication. PMID- 6273953 TI - Structure and function of plasmid ColK. PMID- 6273954 TI - Behavior of Inc-Q plasmids in Agrobacterium tumefaciens. PMID- 6273955 TI - Inhibitory effect of some flavonoids and falvonoid mixtures on cyclic AMP phosphodiesterase activity of rat heart. PMID- 6273956 TI - Pharmacological activity of leukotrienes A4, B4, C4 and D4 on selected guinea pig, rat, rabbit and human smooth muscles. AB - The myotropic activity of leukotrienes A4, B4, C4, D4 and histamine has been evaluated on selected smooth muscle preparations. LTA4, B4, C4 and D4 were several times more potent than histamine on the guinea-pig lung parenchymal strip, while on the guinea-pig trachea, LTB4 was less active. The guinea-pig ileum either in segments or in strips of longitudinal muscles responses well to LTC4, LTD4 and histamine but not to LTA4 and LTB4. Rat and rabbit lung parenchymal strip showed very little sensitivity for leukotrienes whereas human parenchymal strips and bronchi were nearly as sensitive as the guinea-pig lung. PMID- 6273957 TI - Refractoriness of diabetic platelets to inhibitory prostaglandins. AB - Inhibition of collagen-induced platelet aggregation by either endothelial extracts, prostacyclin, prostaglandin E1 or prostaglandin D2 was investigated. The inhibition was less efficient with diabetic platelets than with platelets from normal donors. The refractoriness of diabetic platelets to inhibitory prostaglandins was observed both with platelet-rich plasma and platelets isolated from their plasma. Moreover levels of cyclic AMP in resting platelets and after stimulation by either PGE1 or PGD2 were lower in diabetic platelets than in normal platelets. It is concluded that the weaker response of diabetic platelets to inhibitory prostaglandins could be related to their content in cyclic AMP. PMID- 6273959 TI - Opiate and benzodiazepine receptors. PMID- 6273958 TI - [The endorphin system and mental diseases in the light of the hedonistic theory (author's transl)]. PMID- 6273960 TI - Effects of increased blood ammonia concentrations on the concentrations of some metabolites in rat tissues. AB - Rats were given ammonium chloride intravenously (5 and 10 mg per 100 g body weight) and sacrificed 30 sec, 5 min and 30 min after injection. Ammonia concentrations were determined in brain, liver, skeletal muscle and epididymial fat, and glycogen, glucose, pyruvate, lactate, ATP and cyclic AMP were assayed in brain, liver and muscle. In control animals, muscle and fat showed a high capacity for ammonia accumulation. After injection of NH4Cl the ammonia concentration increased in all tissues analysed, particularly in muscle and fat, where ammonia concentrations remained elevated for 30 min. In brain, glycogen was reduced whereas glucose, pyruvate, lactate and cyclic AMP levels were raised. In liver, glycogen was lower and glucose higher than in controls. ATP concentrations were unchanged in both tissues. In muscle samples taken 30 sec and 5 min after dosing, glycogen concentrations were not affected but glucose, pyruvate and lactate concentrations were reduced and ATP concentrations were increased. In muscle samples taken after 30 min metabolite concentrations were normal again but glycogen concentrations had dropped. PMID- 6273961 TI - Radiosensitization of Pseudomonas radiora O-1 by N2O in aqueous suspension. PMID- 6273962 TI - [Effect of cAMP accumulation activators on the individual stages of gene expression in cells in acute body radiation lesion. 2. The effect of serotonin and papaverine on the chromatin matrix activity of rat liver and spleen cell nuclei in the early periods after total-body x-ray irradiation of the animals]. PMID- 6273964 TI - [Main directions of glycoprotein breakdown in gamma-irradiated aqueous solutions]. PMID- 6273963 TI - [Role of beta-adrenoreceptors in the radioprotective effect of isoproterenol]. PMID- 6273965 TI - Roentgenologic diagnosis of primary corticotropin-producing carcinoid tumors of the mediastinum. AB - A study was undertaken of five patients with Cushing syndrome due to adrenocorticotropin (ACTH) production by thymic carcinoid tumor (primary mediastinal APUDomas), including two recent patients examined by computed tomography (CT) of the chest. Plain roentgenography of the chest initially failed to detect tumor in four of the five patients, while CT of the chest yielded definitive diagnostic information in both patients in whom it was employed. For one of these patients, a mediastinal tumor could be seen retrospectively on plain roentgenograms of the chest, although it had been missed on the first examination. One of the tumors appeared to be partially calcified on CT scan, a finding not previously reported. Blastic osseous metastasis, which is common when malignant carcinoid tumors spread to bone, was seen in one patient. Our data suggest that in patients with suspected ectopic ACTH production, CT scanning of the mediastinum should be performed early in order to avoid delay in diagnosis of an ACTH-secreting carcinoid tumor of the mediastinum. PMID- 6273966 TI - Radionuclide angiography for assessment of hyperthyroidism. PMID- 6273967 TI - The role of mammography in evaluating patients with early carcinoma of the breast for tylectomy and radiation therapy. AB - The value of mammography in treatment planning for early breast cancer following excisional biopsy was studied in 38 patients. One-third of the post-biopsy mammograms yielded information useful for therapy or follow-up, specifically the presence or absence of gross residual tumor [11] and detection of occult lesions in the opposite breast or axillary nodes [3]. They also proved valuable as base line studies, since scars seen on post-therapy mammograms may simulate new or recurrent tumor [3]. Approximately half [21] of the post-biopsy studies were nondiagnostic because of dense parenchyma [14] or biopsy-related distortion [7]. Four were considered falsely positive for residual tumor. These results indicate that mammography can contribute significantly to the management of patients undergoing tylectomy and primary radiotherapy for breast cancer. PMID- 6273968 TI - Biliary excretion of drugs and other xenobiotics. PMID- 6273969 TI - Endogenous opioid peptides--the enkephalins and endorphins. PMID- 6273971 TI - [Cellular substrate for the src gene product (author's transl)]. PMID- 6273970 TI - Inhibitors of angiotensin-converting enzyme. PMID- 6273972 TI - [Biochemical mechanism of signal transduction (author's transl)]. PMID- 6273973 TI - [Response of the oral mucosa in the rat to various suture materials most frequently used in periodontal surgery]. PMID- 6273974 TI - [Bone metastases of stomach cancer. Apropos of 7 cases]. PMID- 6273975 TI - [Milking and milk production in sedentary village herds in the north of the Ivory Coast]. PMID- 6273976 TI - [Cycles of biomass and regrowth following cutting of savannah in the Ivory Coast]. PMID- 6273977 TI - Properties of mevalonate-activating enzymes in developing chick brain. AB - Phosphorylation of mevalonic acid has been studied in 1-3 day chicks. Addition of Mg2+ or Mn2+ strongly increased the formation of phosphorylated derivatives. Mevalonate phosphorylation showed an absolute nucleotide requirement, being ATP and ITP the most effective phosphate donors. Unlike other vertebrate mevalonate kinases, the enzyme from chick brain did not require thiol group protectors as activators. However, the enzyme was found to be sensitive to thiol binding reagents. PMID- 6273978 TI - [Localized thermotherapy in malignant tumors. Preliminary clinical evaluation]. PMID- 6273979 TI - [Human cytomegalovirus]. PMID- 6273980 TI - Possible mechanism of action of piperazine derivatives on liver mitochondria. I- Effect of p-toluyl m-nitro-piperazine (p-TNP). AB - The effect of p-toluyl m-nitro-piperazine on energy conservation processes in rat liver mitochondria is presented. The drug showed an inhibitory effect on the three segments of the respiratory chain and on the ATPase system. NADH oxidase and NADH dehydrogenase activity was inhibited 100%. The velocity and amplitude of swelling induced by glutamate, succinate, ascorbate + TMPD, and ATP was significantly changed by p-toluyl m-nitro-piperazine. It was suggested that the general action of the drug on mitochondrial metabolism would be concerning with modifications on mitochondrial membrane. PMID- 6273981 TI - HPLC analysis of erythrocyte pyrimidine-5'-nucleotidase inhibition by lead. AB - The activity of pyrimidine-5'-nucleotidase (P5N) (EC 3.1.3.5) was assayed in microsamples of rat blood using high performance liquid chromotography (HPLC). The assay is based on the measurement of enzymatically formed uridine in erythrocyte hemolysates (10-50 microliter) and produces a linear activity curve from 5 to 1000 microM/g Hb/h. Acute (22 h) administration of lead acetate ip to rats induced a dose-dependent inhibition of P5N activity in erythrocyte samples. PMID- 6273982 TI - Isolation of viruses from outbreaks of suspected tenosynovitis (viral arthritis) in chickens. AB - Specimens from the legs of chickens from 84 outbreaks of suspected tenosynovitis were examined for the presence of viruses by culture in chick embryo lung or liver cell monolayers. All samples were from broilers or broiler breeders, ranging in age from dead-in-shell embryos to 36 weeks old. Twenty-five outbreaks (29.8 per cent) yielded viruses of which 12 were reoviruses alone, 12 adenoviruses alone, and one, a mixture of both types of virus. Rupture of the gastrocnemius tendon was seen in 12 outbreaks and viruses were isolated from six of these: three were reoviruses and three were adenoviruses. Approximately half the affected flocks from which specimens were received were in the six to 14 week age range. With one exception, all the reovirus isolations were made from chickens of 11 weeks or under, while adenovirus isolations showed more scatter with regard to age. PMID- 6273983 TI - Influence of temperature of incubation on chicken embryo tracheal organ cultures and chick embryos infected with strains of avian infectious bronchitis virus. AB - The ciliostatic and embryo lethal effect of four strains of avian infectious bronchitis virus (IBV) were observed at two different incubation temperatures. In organ cultures, nephrosis-producing IBV strains (GN-2 and M-41) were slightly more ciliostatic and respiratory strains (Beaudette and Kita-1/Tokushima) less ciliostatic at 40.5 degrees C than at 37 degrees C. Mortality of chicken embryos infected with the GN-2, M-41 and Kita-1/Tokushima strains was greater at 40.5 degrees C than at 37 degrees C, while the reverse was true of the Beaudette strain which caused no embryo mortality at 40.5 degrees C. The Beaudette strain of IBV was the only one which could not be recovered from the kidneys of young chickens four days after infection. These results indicate that the different IBV strains may show variation in virulence at different temperatures in ovo and in organ cultures and this may be a reflection of the virulence and tropism for chickens. PMID- 6273984 TI - Influence of environmental factors upon the survival of Aujeszky's disease virus. AB - The survival of Aujeszky's disease (pseudorabies) virus outside the living host was found to be dependent on pH level and temperature. Virus inactivation occurred at a rate varying from 0.04 log10 per day at 4 degrees C to 0.6 log10 per day at 37 degrees C at the optimum pH levels of 6 to 8. Fluctuation of the temperature between 4 degrees C and 37 degrees C had no apparent effect upon the titre of the virus. At a steady temperature of -13 degrees C, the virus was rapidly inactivated at all tested pH levels. For long-term storage it is recommended that the pH should be adjusted to between 6 and 7, that suspensions should be kept at -90 degrees C and that freezing and thawing should be rapid so as to pass through the just subzero temperature zone as quickly as possible. Drying, both on glass and on gelatin, caused inactivation of the virus at all pH levels, as did exposure to ultraviolet light. PMID- 6273985 TI - [Incidence of ornithosis-psittacosis and herpetic infections in carrier pigeons and psittacines in Belgium]. PMID- 6273986 TI - [Current trends in the antimicrobial therapy of chronic bronchitis]. PMID- 6273988 TI - [Failures of standardized treatments in pulmonary tuberculosis. Possibilities of prevention and therapeutic recovery]. AB - In view of a better knowledge of therapeutic failures in operational conditions four groups of patients were analyzed, which included a total of 1511 patients, both new cases and patients re-admitted for tuberculosis of the pulmonary apparatus, that had been treated according to standardized regimens. In three of these groups failures had been recorded at a rate of 3,9% intra-therapeutic failures and 7,6% post-therapeutic ones in the first 18 months after completing the therapy. Out of the four groups a lot was selected, which included 127 cases of therapy failure. This lot was studied under the aspect of causes of the failure and of the possibilities for recovery. The major causes of the failure were non-cooperation of patients (in 48% of the cases), the initial severity of the lesions, associated diseases, the re-admittance, the secondary resistance. Recovery of the intra-therapeutic failures was possible in 46% of the cases, and of patients with post-therapeutic failure in 72% of the cases. On the basis of an analysis of risk factors a series of proposals are made for preventing such failures. PMID- 6273987 TI - [Attitude toward patients with pulmonary pathological images supposed to be tuberculosis but without bacteriological confirmation]. AB - In view of establishing an attitude toward patients with pathological pulmonary images, suspected of tuberculosis but without bacteriological confirmation, a study was organized in an area serviced by 9 offices of phthisiology. A total of 154 such cases were identified between 1st January and 30th April 1978. These were followed up and controlled by radiological as well as bacteriological examinations for a period of one year. The anti-tuberculous treatment did not have to be applied except when the diagnosis was confirmed as tuberculosis. In 11 of the patients (7,1%) the diagnosis was made from the start, and it was established that they had active, non-bacillary pulmonary tuberculosis. In another 49 patients (31,8%) the suspicion was abandoned, and tuberculosis. In another 49 patients (31,8%) the suspicion was abandoned, and various other pulmonary affections, but not tuberculosis, were diagnosed, either of chronic or acute type. The remaining 94 patients (61,1%) remained suspected of tuberculosis. In 16 of them, in spite of the provisions made in the protocol of the study, anti tuberculous treatment was applied. Of them 3 died of pulmonary cancers during the follow-up. The remaining suspects were not treated. Five of them (7,3%) have developed active tuberculosis, bacteriologically confirmed in 3 of the cases. The rest had a normal eventless evolution. The authors consider the careful investigation and follow-up of suspects for a period of one year as the most correct attitude in such cases. PMID- 6273990 TI - [Effectiveness of abbreviated biphasic treatment in pulmonary tuberculosis]. PMID- 6273989 TI - [Post-tuberculosis syndromes appearing in cases undergoing initial chemotherapy]. AB - The study of frequency and of the nature of post-tuberculous syndromes in a group including 1351 patients with pulmonary tuberculosis at their first treatment, investigated between 1974 and 1977, has demonstrated that in the present stage of initial chemotherapy with standardized drug regimens, the post-tuberculous syndromes are decreasing in most of the cases, developing in those of the patients that presented with advanced froms of the disease at the start of the the therapy (2,8% in our study). At the same time one should note the prevalence of patients that have benefited from initial courses of chemotherapy with classical and relay drugs, such patients usually having less severe anatomic and radiologic lesions. The study of types of post-tuberculous syndromes has revealed the increased frequency of the bronchitic syndrome. PMID- 6273991 TI - [Results of 6-month and 9-month treatments with rifampin, isoniazid and ethambutol in cases of newly detected tuberculosis]. PMID- 6273992 TI - [Results of bronchoscopic examinations of tuberculosis patients undergoing complex chemotherapy]. PMID- 6273993 TI - [Integrated system of anti-tuberculosis supervision of several groups of high risk populations]. PMID- 6273994 TI - Continuous intra-arterial pH-monitoring using monocrystalline antimony as sensor. A study in non-heparinized dogs. AB - Continuous intra-arterial pH-monitoring was performed on anaesthetized and artificially ventilated dogs. The pH sensor consisted of monocrystalline antimony. The sensor was placed on the tip of a catheter, length approximately 35 cm and diameter 0.8 mm. No anticoagulation was undertaken. The pH range studied was 7.01-7.53, the pH being changed by infusion of sodium bicarbonate ro ammonium chloride. Arterial reference blood samples for in vitro blood gas analyses were taken under stable monitoring conditions and analysed with a minimal delay. The millivolt signal from the antimony sensor was found to be a linear function of the pH of the reference blood samples in the intra-arterial pH range studied. The sensitivities of the different antimony sensors were closely correlated but not identical. When the antimony sensor was used completely uncalibrated, the standard deviation of the registered potential corresponds to 0.06 pH units. After single point calibration a standard deviation of 0.03 pH units was obtained. These standard deviations include a suspected influence of minor changes in the partial pressure of oxygen during the experiments. PMID- 6273995 TI - Angiotensin-converting enzyme in acute myocardial infarction and angina pectoris. AB - Serum activity of angiotensin-converting enzyme was measured by serial analysis in 19 patients with acute myocardial infarction and in eight patients with angina pectoris. As a rule no changes in enzyme activity occurred during 6 days observations. However, two patients with infarction exhibited a pronounced fall of enzyme activity which could not be related to clinical events. The analysis seems to have no place in the diagnosis and management of patients with myocardial infarction. PMID- 6273996 TI - Human adipose tissue blood flow during prolonged exercise, III. Effect of beta adrenergic blockade, nicotinic acid and glucose infusion. AB - Subcutaneous adipose tissue blood flow (ATBF) was measured in six male subjects by the 133Xe-washout technique during 3-4 h of exercise at a work load corresponding to an oxygen uptake of about 1.71/min. The measurements were done during control conditions, during blockade of lipolysis by nicotinic acid, during acute i.v. beta-adrenergic blockade by propranolol, and during continuous i.v. infusion of glucose. The most pronounced lipid mobilization and utilization during work was seen in the control experiments where ATBF rose 3-fold on average from the initial rest period to the third hour of work. No increase in lipolysis and no increase in ATBF were found when lipolysis was blocked by nicotinic acid (0.3 g/h). Propranolol treatment (0.15 mg/kg) reduced lipolysis and nearly abolished the increase in ATBF during exercise. Intravenous administration of glucose (about 0.25 g/min) did not influence lipid metabolism (evaluated by the respiratory quotient) nor did it reduce the ATBF response to exercise. These results are inconsistent with the hypothesis that increase in ATBF during exercise is elicited via direct stimulation of vascular beta1-receptors, while they are not in disagreement with the hypothesis that adipose tissue vasodilation during exercise is secondary to metabolic events connected to lipolysis. PMID- 6273997 TI - Nesidiodysplasia and nesidioblastosis of infancy: ultrastructural and immunohistochemical analysis of islet cell alterations with and without associated hyperinsulinaemic hypoglycaemia. AB - The pancreata of hyperinsulinaemic, hypoglycaemic infants, having the anatomic anomalies characterised as "nesidioblastosis', were compared with age matched control infants by light microscopy, and electron microscopy. While the hyperinsulinaemic infants showed an apparent increase in total endocrine volume compared with control infants, the light microscopic alterations of topographic maldistribution of endocrine cells, irregularly defined islets and intermingling of endocrine with exocrine elements were common to both hypoglycaemic and control groups. In at least one control case, the total endocrine cell volume was comparable to that seen in hypoglycaemic infants and was not associated with endocrine dysfunction. The ratios of insulin: somatostatin: glucagon cells in hypoglycaemic infants were similar to those of control infants. The results of both immunohistochemistry and electron microscopy gave strong indications of endocrine cells containing more than one immunoreactive peptide and heterogenous granule populations respectively. Ultrastructurally, "composite' cells with features of both exocrine and endocrine differentiation were found with some frequency in two hypoglycaemic infants. These findings are discussed in the light of current notions of gastroentero-pancreatic endocrine system development. We conclude that "nesidioblastosis' as currently defined is not the anatomic substratum of infantile hyperinsulaemic hypoglycaemia. We propose the term "nesidiodysplasia' to encompass the apparently increased and possibly maldistributed and/or malregulated endocrine cells associated with the clinical manifestations of hyperinsulinaemia. The precise relationship between the presumed anatomic abnormalities and abnormal insulin secretion remains to be clarified by further investigations. PMID- 6273998 TI - Scanning electron microscopy of the Wistar rat colon. PMID- 6273999 TI - Dynamics of the interaction between Entamoeba histolytica and components of the immune response. III. Fate of antibodies after binding to the cell surface. AB - Binding to the surface of living E. histolytica trophozoites of 125I-labelled anti-amoeba antibodies or of such antibodies and antiglobulin resulted in a rapid disappearance of the antibody into the medium. This disappearance consisted of a rapid temperature-independent and a subsequent slow temperature-dependent phase. The disappearance of antibody from fixed trophozoites merely consisted of a temperature-independent rapid phase whereas no slow phase was observed. The magnitude of the rapid phase of antibody disappearance was virtually the same in living and fixed cells. During culture of living trophozoites for 4 h the amount of iodine-labelled material sedimenting lighter than monomeric IgG increased from 2% to 14% of all labelled material in the case of the single antibody layer. The corresponding values for the double layer were 2% and 32%. Of the material lighter than IgG, 10% of that in the single layer and 26% of that in the double layer were detected in the medium. Thus, during the 4-h culture period 25% of the originally amoeba-bound single layer of antibody probably disappeared from the cells not complexed with antigen, 27% disappeared as complexes heavier than IgG, 10% disappeared as material lighter than IgG, and 38% persisted on or within the cells. The corresponding values for the double layer of antibody were 10%, 30%, 26% and 34%. PMID- 6274000 TI - Intrauterine varicella: a report of two cases associated with hyper-A immunoglobulinemia. AB - Varicella during pregnancy may occasionally harm the fetus. This report described one case of congenital varicella syndrome and another of infantile herpes zoster after maternal varicella in the 16th and 29th week of pregnancy, respectively. A remarkable finding was that both mothers and infants had raised levels of serum IgA. This suggests that varicella is prone to lead to congenital disease in infants of pregnant women with an immunologic, perhaps genetically determined abnormality. PMID- 6274001 TI - Current status of Pseudomonas infections and antibiotics. PMID- 6274002 TI - Phosphate metabolism in renal stone formers. (II): Relation to renal tubular functions and calcium metabolism. AB - Patients with idiopathic recurrent calcium nephrolithiasis (n = 57) and controls (n = 16) were investigated regarding the relationship between renal phosphate handling, other renal tubular functions and calcium metabolism. Incomplete renal tubular acidosis (RTA) was disclosed in 13 patients. RTA patients together with stone formers with normal renal acidification capacity (SF) exhibited low values for serum phosphate and renal threshold phosphate concentration (TmP/GFR) compared with controls. TmP/GFR was lower in RTA patients than in stone formers with normal renal acidification. Hypercalciuria of the absorptive type with normal serum PTH and urinary cAMP concentrations was a common finding in both stone patient groups, whereas no patient displayed unequivocal evidence of parathyroid hyperfunction. Fractional excretion of sodium was raised in both SF and RTA patients compared with controls. There was a positive relationship between the fractional excretion of phosphate and sodium in all subjects as a group. TmP/GFR was negatively correlated to fractional excretion of sodium. Twenty-three percent of RTA patients and 8% of SF displayed tubular proteinuria which often was associated with low TmP/GFR levels and enhanced natriuresis. It is concluded that a defective renal tubular phosphate handling is common in calcium stone formers and often associated with signs of other tubular dysfunctions. The altered phosphate handling seems to be unrelated to hypercalciuria. PMID- 6274003 TI - Phosphate metabolism and renal calcium stone disease. AB - The purpose of the present investigation was to study various aspects of phosphate metabolism in renal calcium stone patients with special reference to the renal handling of phosphate and its relationship to other renal tubular functions and calcium metabolism. Serum phosphate and the capacity for renal tubular reabsorption of phosphate were lower in stone patients than in controls and decreased with advancing age. Reduced tubular phosphate reabsorption was particularly evident in stone patients with other tubular dysfunctions. Absorptive hypercalciuria was common, but unrelated to the renal tubular reabsorption of phosphate. Parathyroid hyperfunction was not observed in stone formers. Various loads of dietary phosphate resulted in similar renal adaptive responses in controls and stone formers. Orthophosphate supplementation had metabolic consequences with potentially beneficial effects for stone prevention (increased urinary pyrophosphate, decreased urinary calcium). The altered renal handling of phosphate in calcium stone formers may reflect a primary (independent of parathyroid hormone) renal tubular dysfunction in phosphate reabsorption. PMID- 6274004 TI - Analysis of inorganic fiber concentrations in biological samples by scanning electron microscopy. AB - Analyzing fibers with electron microscopic techniques involves several preparation steps, especially during the analysis of fibers in human tissue. The influence of these steps on the analytical result was studied in detail. Fiber number was unaffected by the mild sonication of fiber suspensions analyzed with scanning electron microscopy. Significant fiber losses appeared during the filtration of fiber suspensions through 0.8-micrometer pore-size Nuclepore membranes. Shrinkage of the tissue during dry ashing broke lung fibers and consequently increased the fiber concentration and affected the fiber length distribution. Dry ashing, however, removed more of the organic debris than wet ashing; thus specimens prepared with dry ashing were more suitable for scanning electron microscopic analysis. A fairy good correlation was demonstrated for the analysis of fibers with scanning and transmission electron microscopy. PMID- 6274005 TI - [Oral contraception and cancer risk]. AB - Oral contraception was introduced twenty years ago and is today the most popular method of preventing pregnancy. Some undesirable side effects of these drugs are well known, while others are only suspected. Many investigations have been undertaken to elucidate whether or not the Pill has carcinogenic properties, and the present study describes the current state of knowledge on this question. At present no evidence exists to suggest that oral contraceptives have carcinogenic effects on the human breast or genital organs. A casual relationship between use of these drugs and development of liver tumors is possible but far from proven. PMID- 6274006 TI - [Secondary tumors: a case contribution on the formation of distant metastases in the jaw bones]. PMID- 6274007 TI - Natural equine viral arteritis in foals. PMID- 6274008 TI - [Control of sodium-potassium pump on proliferation of mammalian cells (author's transl)]. PMID- 6274009 TI - [Cyclic CMP (author's transl)]. PMID- 6274010 TI - [Present status of research on narcotic receptors (author's transl)]. PMID- 6274011 TI - [The endogenous analgesia system (author's transl)]. PMID- 6274012 TI - [Nerve growth factor: its distribution, chemistry and function (author's transl)]. PMID- 6274013 TI - [On the requirements and dietary allowances of carbohydrates, fibers and trace elements (author's transl)]. PMID- 6274014 TI - [Mechanism of GABA potentiation by benzodiazepines (author's transl)]. PMID- 6274015 TI - Amoebiasis as a cause of death in a Scottish resident. AB - An elderly woman, resident throughout her life in Scotland presented with abdominal pain and diarrhoea. A provisional diagnosis of ulcerative colitis was made on sigmoidoscopy. At post mortem Entamoeba Histolytica was demonstrated in considerable numbers in the mucosal ulcers. It would appear though extremely rare that amoebiasis should be considered in the differential diagnosis of colonic ulceration. PMID- 6274016 TI - Gonadotropin-releasing hormone: regulation of adenosine 3',5'-monophosphate in ovarian granulosa cells. AB - The antigonadal effects of gonadotropin-releasing hormone in ovarian granulosa cells are due to attenuation of the adenosine 3',5'-monophosphate (cyclic AMP) response to follicle-stimulating hormone. Agonists of gonadotropin-releasing hormone progressively inhibit adenylate cyclase and stimulate phosphodiesterase activities in cultured granulosa cells, indicating that blockade of gonadotropin action is attributable to the combined effects of decreased production and increased degradation of cyclic AMP. PMID- 6274017 TI - Regulation of muscle differentiation: stimulation of myoblast fusion in vitro by catecholamines. AB - Epinephrine and isoproterenol provoke primary chick myoblasts to initiate precocious cell fusion. Both the rise in intracellular adenosine 3' ,5 monophosphate (cyclic AMP) and cell fusion generated by these effectors are prevented by propranolol, which is a specific blocker of the beta-adrenergic receptor. Propranolol has no effect either on the precocious cell fusion provoked by prostaglandin E or on cell fusion in control cultures. The results support the idea that a rise in cyclic AMP is the critical intracellular change responsible for initiating events that culminate in myoblast differentiation 4 to 5 hours later. They also indicate that the culminate in myoblast differentiation 4 to 5 hours later. They also indicate that the hormone responsible for the positive regulation of myoblast differentiation in vitro is not acting through the beta adrenergic receptor. PMID- 6274018 TI - Angiotensin II binding sites in rat brain. PMID- 6274019 TI - Reversal of induced ischemic neurologic deficit in gerbils by the opiate antagonist naloxone. PMID- 6274020 TI - Epinephrine reduction of heme: implication for understanding the transmission of an agonist stimulus. AB - Alpha-adrenergic agonists that promote platelet aggregation were found to reduce ferric heme to ferrous heme. Agents that bind iron in heme inhibited epinephrine induced platelet aggregation. It is proposed that epinephrine first binds to its receptor and then reduces an adjacent heme group to transmit its agonist stimulus. PMID- 6274021 TI - Group B erythrocytes enzymatically converted to group O survive normally in A, B, and O individuals. AB - With an alpha-galactosidase, B erythrocytes can be converted to blood group O under conditions that neither impair their viability in vitro nor affect their ability to survive normally after transfusion to individuals of groups O, A, and B. Such an approach has the potential for producing enzymatically converted group O cells for use in transfusion therapy. It should also be possible to convert A cells to group O by using the appropriate alpha-N-acetylgalactosaminidase. PMID- 6274022 TI - Stereoisomers of N-allylnormetazocine: phencyclidine-like behavioral effects in squirrel monkeys and rats. AB - (+/-)-N-Allylnormetazocine is a benzomorphan opioid with psychotomimetic effects. The pure stereoisomers of this compound, as well as the racemic mixture, were compared to phencyclidine for their behavioral effects on squirrel monkeys and rats trained to discriminate phencyclidine from saline. Dose-response determinations were made for responses to phencyclidine, to a racemic mixture of N-allylnormetazocine, and to the pure levo and dextro isomers of N allylnormetazocine. In both rats and monkeys, the dextro isomer and the racemic mixture produced dose-dependent responses appropriate for phencyclidine; the levo isomer did not produce the responses appropriate for phencyclidine at any of the doses tested. In both species, the levo isomer was more potent than the dextro isomer in decreasing the rate of responding. Thus racemic N-allylnormetazocine is a mixture of compounds that produce different behavioral effects. PMID- 6274023 TI - Specific chromosome defect associated with human small-cell lung cancer; deletion 3p(14-23). AB - A specific, acquired chromosomal abnormality (deletion 3p) has been found in at least one chromosome 3 in 100 percent of the metaphases in 12 of 12 cell lines cultured from human small-cell lung cancer tissue and in 2-day tumor culture specimens from three patients. Analysis of the shortest region of overlap shows the deletion to be 3p(14-23). This specific change was not seen in five of five lung cancer cell lines other than small-cell lung cancer or in two lymphoblastoid lines cultured from cells of small-cell lung cancer patients whose tumors had the 3p deletion. PMID- 6274024 TI - [Modern aspects in management of psychotic children (author's transl)]. AB - The idea of hope is paramount in the management of psychotic children. The different alternatives were developed with reference to the modern conceptions of the psychoanalytical theory. Day hospitals, each of which admits children from a given district, seem to allow optimal active therapy. Management includes: individual psychoanalysis for the child; life groups to which the child belongs during the day; a consistent institutional policy based on psychoanalytical theories; leaving the child with it's own or an auxiliary family and supporting this family; minimal chemotherapy. The authors report major improvement or recovery in one-third to one-half of the children. PMID- 6274025 TI - [Questioning in child psychosis (author's transl)]. AB - Case study of infantile psychosis characterized by serious behavioral and language disorders, the most conspicuous feature being the unceasing questioning, giving evidence of missing essential references, ambiguous mode of communication and consisting of endless, sometimes metaphysical questions, usually occulted by the child when grown to "Years of reason". PMID- 6274026 TI - [Gifted children. Psychological and psychopathological aspects (author's transl)]. AB - After having tried to define the term "gifted children" the authors record the principal works devoted to this subject and the opinions, often controversial raised by the attitude in regard to exceptional children in different countries. Then, are given the means to detect gifted children as well as the characteristics in the recognition of the very young gifted children. In a third section are described the psychology and the difficulties of adaptation which these children sometimes encounter (scholastic problems, behavioral and personality problems, seldom psychotic). Finally, the orientation of gifted children can be undertaken in three ways, all of which present advantages and disadvantages: grouping in special classes, speeding up and enrichment of the programmes. PMID- 6274027 TI - [A seasonal neurologic disease: paralysis following tick-bites (author's transl)]. AB - Seasonal inflammatory neurologic disorders following tick-bites have been reported more and more often during the last few years. They usually consist in a peripheral neurologic syndrome. However, involvement of the central nervous system is not uncommon. Two new cases seen at the Val-de-Grace Hospital are reported here. The author reviews the circumstances in which the disease occurs and it's clinical features. The course is variable and may result in serious sequelae. Pathogenesis remains unknown. Viral, toxinic, and immuno-allergic factors have been incriminated. Practitioners should consider this diagnosis, during the summertime, especially in high-risk subjects (campers, agricultural workers, troops during field exercise). PMID- 6274028 TI - [Present data of Fahr's syndrome. Report of two cases of idiopathic strio-pallido dentate calcifications revealed by computerized tomography (author's transl)]. AB - The authors report two computed tomographic observations of Fahr's disease without disturbances in calcium-phosphorus metabolism; the first case is a by chance discovery and the second one is characterized by a cerebellar tremor. From the literature data they discuss successively clinical, radiological and computed tomographic, neuropathological, aetiopathogenetic and therapeutic aspects actually presented by strio-pallido-dentate calcifications (SPD) or Fahr's disease. PMID- 6274029 TI - [Psychological profile with Cushing disease (author's transl)]. AB - Fifty-six patients with clinically and biologically unequivocal Cushing disease underwent psychological evaluation including at least one interview, a graphological test, a tree test, and a Szondi test. Patients were found to be hyperadapted, with a rigid repressive system, concealing major anxiety. These patients are closely dependent on their familial and professional affective relationships. This probably establishes the intensity of their repressive system. They need approval, appreciation, and recognition. Obsessional features are found (scrupulous conscience, compulsive activity) as well as manic depressive features (fluctuations between depression and euphoric activity acting as a defense against depression). During the interviews it was established that onset of Cushing disease occurred a few months after prolonged stress; this stress dealt electively with the patients affective relationships. PMID- 6274030 TI - [Chronological study of signs of myoclonic encephalopathy in hemodialysis patients (author's transl)]. AB - Twelve patients with chronic renal failure, on maintenance hemodialysis have a stereotyped illness. The dialysate was made up from a high level aluminium tap water. After having been on haemodialysis for 10 to 12 months, they suffer from more and more acute osteodystrophic, osteomalacic symptoms. Then, distinctive EEG abnormalities appear. These first symptoms allow an early diagnosis of the progressive dialytic encephalopathy. The neurological symptoms appear some months later. Eight patients died from this encephalopathy. Four patients are still alive and are, from at least two years, on maintenance haemodialysis with a free aluminium water. In these 4 cases, the evolution of the disease is good: osteomalacic symptoms disappear in a year; the neurological symptoms are still present though transient. EEG abnormalities remain and blood aluminium level is about 100 micrograms/L. These cases show, once more, the significant role of aluminium as an aetiological factor in "progressive dialysis encephalopathy". PMID- 6274031 TI - [Is there a treatment for paranoia? (author's transl)]. AB - Is there a treatment for paranoia? Our answer is yes. A dual sensitive relationship with the patient must be avoided. The ties which link the patient to his practitioner and to the other therapists involved must be supple and diversified. Chemotherapy must be warranted and accepted by the patient. Among the psychopharmaceutical agents, anisamides are particularly helpful in improving social integration. PMID- 6274032 TI - [Tiapride in the treatment of neurological and psychiatric disorders in the elderly (author's transl)]. AB - One hundred patients, aged between 60 and 92 years, were treated with tiapride for neurological disorders (abnormal movements, buccofacial dyskinesias, dopa therapy complications, ballism, eyelid tics, senile tremor, post-traumatic headache, delirium tremens), psychiatric disorders with more or less marked agitation and of various types (hysteria, depression, mood disturbances, hypochondria, delusions, hallucinations), or for mental deficiency, senile dementia, or arteriopathic dementia. Results were excellent, being satisfactory in 70 p. cent, and even more marked in some groups. Tolerance was very good, with some rare cases of somnolence. The efficacy and safety of tiapride makes it of particular value for treating neuropsychiatric disorders in geriatric patients. PMID- 6274033 TI - [Anorexia nervosa: protest and depression (author's transl)]. AB - Diagnosis of anorexia nervosa follows strict criteria. It's most striking manifestations are somatic and may be suggestive of other conditions. It originates in a psychological situation which is characteristic of the illness. A troubled relationship between mother and child in the past is usually demonstrated during interview. Once anorexia nervosa has become apparent, complex problems arise between patient and family. These problems differ according to the patient's sex. The best management can be inferred from these facts. Physical and psychiatric care should be combined. Psychiatric management ranges from family therapy (which may be questionable) to longterm individual psychotherapy. PMID- 6274034 TI - [Sultopride in acute manic psychoses (author's transl)]. AB - Sultopride, a benzamide derivative, provides rapid sedation. This makes it one of the main drugs used in psychiatric emergencies. Our study includes 26 cases of psychomotor hyperactivity originating in manic-depressive psychosis of schizophrenia. The medication had no effect on wakefulness. It was always effective on agitation but not on delusion or hallucinations. Manic forms of primary delirious experiences are the best indication for sultopride. It is mainly effective on delirium as a state rather than on delirium as an idea (Wahn, Delusion) which is secondary from a pathogenetic standpoint. PMID- 6274035 TI - [The alcoholic therapeutic strategy in an internal medicine unit (author's transl)]. AB - The authors, in an Internal Medicine and therapeutic Unit (St Andre's hospital Bordeaux) starting from a psychosomatic approach of the patients, studied the post hospital care alcoholic's reinstatement. Referring to their hospital experience (medical social and political) collating with the pathological polymorphism which faces up to the therapeutic fragmentation, they tried to offer a therapeutic strategy which makes responsible all those who possibly draw near the alcoholic, informs in a global and unexclusive step, even including a new process: the family therapy. PMID- 6274036 TI - [Masked depression and paucisymptomatic depression (author's transl)]. AB - All depressive syndromes include physical as well as psychological features. Physical symptoms in major "endogenous" depression are well known. In other forms, called masked depressions, various physical disorders are the patients main complaint and may be misleading. Localized pain and paresthesia are common. Behaviour disorders may mask depression in adolescents. Mood disturbances (loss of interest, anhedonia) as well as the personal and family histories, should be precisely assessed in order to establish diagnosis. Masked depression is not a minor form of depressive syndrome and antidepressant drugs should be used in correct doses over a sufficient period of time. PMID- 6274037 TI - [A major medication for autism: sulpiride (author's transl)]. AB - Sulpiride is a benzamide derivative used in medical practice since 1967. Its original effect became immediately apparent. It is effective against hallucinations as well as against inhibition, but is not sedative. Its antidepressant activity is still controversial. Subsequent studies confirmed the outstanding antiinhibitory effect which leads to better relationships as well as to qualitatively and quantitatively improved vigilance. Side effects, which are minor and reversible, affect the endocrine and nervous systems but do not include cholinergic effects. No drug-dependence is seen with sulpiride. PMID- 6274038 TI - [A medical investigation of 160 chronic alcoholic patients hospitalized in a department of internal medicine. Nature and pathological consequences of the intoxication (author's transl)]. AB - 160 hospitalized chronic alcoholics (124 men and 36 women) were evaluated by the same person. 21 biologic parameters were assessed. A histopathological examination of the liver was performed in 57.5% of the cases. The average daily alcohol consumption (ADAC) during the last five years exceeded 80 g in 140 patients. Results were as follows. Alcoholics usually have a poor socio-economic and professional background and a low educational level, most of them being farmers or workers. The ADAC is 170 g/day for an average duration of 32 +/- 3 years. The diet is unbalanced. The consumption of tobacco is related to age, sex and ADAC. There is no significant correlation between hepatic lesions (cirrhosis: 37 cases, alcoholic hepatitis without cirrhosis: 83 cases, normal liver: 36 cases) and age, sex, diet, ADAC, or the duration of intoxication. Pancreatic involvement was found in 13 patients (6.8%). Neuropsychiatric features were present in 92% of the patients. A significant correlation was found between stellar angiomas, clinical liver involvement, and ADAC, between palmar erythrosis, and clinical liver involvement and between hippocratism and ADAC. VMC and Gamma GT levels are significantly correlated to liver involvement and ADAC. There is a linear correlation between VMC and Gamma GT. The Gamma GT/BUN ratio is significantly correlated to ADAC. No correlation was found between serum vitamin B12, serum folic acid and VMC. No patient was HbS antigen-positive. Anti-HbS antibodies were found in 19.2% of the cases. PMID- 6274039 TI - [Hospital physicians and care of alcoholics (author's transl)]. AB - Hospital physicians should not confine their role to the high cost management of complications resulting from alcoholism. They should also concern themselves with the alcoholic patient himself and take advantage of the opportunity provided by his hospitalization. In each hospital a specific care system for alcoholics should be set up, with facilities for managing eight to fifteen patients. Opening new hospital departments is unnecessary. A relationship should be established with the alcoholic patient. His condition should be recognized and discussed freely. Correct management can only be achieved if sufficient personnel is available: alcoholism specialists, psychologists, secretaries, social workers, etc. Until now the creation of such a staff has usually been refused. PMID- 6274040 TI - [Problems in the treatment of a patient with longstanding epilepsy (author's transl)]. AB - The case of a female patient with longstanding epilepsy is reported. Many problems in achieving a satisfactory therapeutic regimen were encountered. They originated in storage of phenobarbital, which had been prescribed since childhood, interference between phenobarbital, dipropylacetic acid, and phenylhydantoins. Some pharmacokinetic principles are reviewed in the light of this clinical experience. PMID- 6274041 TI - Computed tomography in evaluation of the pancreas. PMID- 6274042 TI - [Models and pragmatism, activation and repetition: reflections on causality of disease in the Senufo people of the Ivory Coast]. PMID- 6274043 TI - Primary scirrhous adenocarcinoma of the colon (signet-ring cell type). PMID- 6274044 TI - Atypical ultrasound findings in gestational trophoblastic disease. PMID- 6274046 TI - [Malignant glomic tumor of the duodenum]. PMID- 6274045 TI - Unusual splenic lesions in two immunosuppressed patients. PMID- 6274047 TI - [Emergency states in certain gastrointestinal tumors]. PMID- 6274048 TI - [Frequency of brain tumors in epileptic patients]. PMID- 6274049 TI - [Myoblastoma granulare tumor ultrastructure]. PMID- 6274050 TI - Active and latent collagenases in rheumatoid synovial fluid with special reference to the pathological state of the joint tissues. PMID- 6274051 TI - Retroperitoneal malignant tumors: Wilms' tumor and neuroblastoma. AB - Wilms' tumor is one of the most curable of all childhood cancers, with a five year survival of approximately 80 per cent. Survival of children with neuroblastoma, on the other hand, is dismal. For both tumors the primary mode of treatment is surgical excision, which should be carried out as an integrated part of multidisciplinary treatment. PMID- 6274052 TI - Natural history of patients after resection of a bronchial carcinoma. PMID- 6274053 TI - [Viral infections in pregnancy]. PMID- 6274055 TI - Calcium lysosomes in rachitic and vitamin D3 replete chick duodenal absorptive cells. AB - Calcium-containing lysomes were described in a previous communication in this series (Davis et al., 1979). Their potential role in intestinal calcium uptake and transcellular transports was hypothesized. To further this notion, the effects of a rachitogenic diet and vitamin D3 repletion were investigated. Intestinal absorptive cells from chicks maintained on a vitamin D deficient diet were characterized by decreased numbers of supranuclear calcium lysosomes. In contrast, intestinal cells from chicks given vitamin D3 (cholecalciferol) subsequent to the rachitogenic diet showed numerous large compound supranuclear calcium lysosomes. Since other steroid hormones are known to effect lysosomes, it is tempting to speculate that vitamin D, itself a steroid hormone, may activate lysosomes which themselves might be involved in calcium homeostatic mechanisms. PMID- 6274054 TI - Circulating catecholamines in exercise and hyperventilation induced asthma. AB - Plasma noradrenaline, adrenaline, and cyclic 3'5' AMP (cAMP) were measured in seven asthmatic patients with known exercise-induced bronchospasm and six matched non-atopic control subjects during a standard treadmill exercise test and then during matched isocapnic hyperventilation. Normal subjects showed a 5.5 fold rise in noradrenaline and a 3.2 fold rise in adrenaline during exercise compared with a 2.1 fold rise in noradrenaline and no significant rise in adrenaline in asthmatics who all developed bronchoconstriction after exercise (mean fall in peak flow rate 28.4 +/- 5.8%). Plasma cAMP rose 1.4 fold in controls but showed no significant rise in asthmatics. This reduced sympatho-adrenal response to exercise in asthmatics is difficult to explain. The failure of circulating catecholamines to rise and stimulate beta adrenoceptors on the mast cell may facilitate the release of bronchoconstrictor mediators. Matched hyperventilation produced bronchospasm in asthmatics (mean fall in peak flow rate 29.0 +/- 4.4%) but no change in catecholamines in either group suggesting that circulating catecholamines have no direct role in exercise-induced bronchospasm but may play a permissive role via the mast cell. PMID- 6274056 TI - Quantitative analyses of specific B cells for sheep red blood cell, phosphorylcholine, and hepatitis B surface antigen in human B cell populations by polyclonal transformation with Epstein-Barr virus. AB - An experimental system was developed to analyse quantitatively specific B cells for various antigens in human B cell populations by polyclonal transformation and immunoglobulin production induced by in vitro infection with Epstein-Barr virus (EBV). EBV-infected cells were cultured in microtest plates for three weeks and specific antibody activities for sheep red blood cell (SRBC) were detected in the culture supernatants by hemolysis and those for phosphorylcholine (PC) and for hepatitis B surface antigen (HBsAg) by passive hemagglutination. It was shown that frequencies of positive cultures with specific antibodies followed one hit type of Poisson's distribution. Large numbers of cultures showed specific antibody activities for SRBC and for PC with tonsillar lymphocytes obtained from four donors. On the other hand, positive cultured with anti-HBsAg antibodies were practically undetectable with three donors. With fourth donor, however, quite a large fraction of cultures showed specific antibody activities for HBsAg. Frequencies of positive cultures with anti-HBsAg antibodies were further analysed with peripheral blood lymphocytes obtained from five donors with serum anti HBsAg. Comparable frequencies of cultures were shown to be positive with anti SRBC antibodies with these donors. The frequencies of cultures with anti-HBsAg antibodies were, however, quite variable among these donors. It was considered that these results directly reflected the sizes of clones with these specificities in human lymphocytes populations obtained from donors with various immunological histories. PMID- 6274058 TI - Scanning electron microscopy of corrosion cast of rat adrenal vasculatures with emphasis on medullary artery under ACTH administration. AB - In the vascular system of the adrenal, the medullary artery is considered to be a shunt which controls cortical function by changing its diameter. To confirm this hypothesis, the medullary arteries of ACTH-treated and control rats were observed by scanning electron microscopy of methacrylate corrosion casts of blood vessels. After ACTH treatment, the medullary arteries were constricted mostly at their distal segments and sometimes also at other portions. This constriction probably results in an effective augmentation of adrenocortical blood flow. The diameter of capillaries in the glomerular zone and medullary sinusoids was markedly increased by ACTH. Small protrusions o the corrosion casts which supposedly corresponded to capillary fenestrations became prominent after ACTH administration in the fascicular and reticular zones. PMID- 6274057 TI - Evidence for adrenergic transmission in the circular smooth muscle of the guinea pig seminal vesicle. AB - Spontaneous contractile activity of the circular smooth muscle of the guinea-pig seminal vesicle consists of twitch-like contractions with a mean frequency of 2.7/min. The amplitude of responses to repetitive stimulation depended on the frequency of stimulation at constant pulse duration and on the pulse durations at constant frequency of stimulation. Mechanical summation in the tension development was observed at low frequency of repetitive stimulation. Adrenaline, noradrenaline and phenylephrine increased the frequency and amplitude of spontaneous contractions. The excitatory actions of adrenaline and noradrenaline were antagonized by phentolamine and phenoxybenzamine. DOPA, metanephrine and normetanephrine were not effective. Tyramine enhanced the spontaneous activity and its action was antagonized by phentolamine. Isoprenaline, at high concentrations, enhanced spontaneous activity, but in the presence of phentolamine, the spontaneous activity was inhibited by isoprenaline and orciprenaline. Isoprenaline inhibited the enhanced spontaneous activity induced by adrenaline, noradrenaline and tyramine. Spontaneous contractions were potentiated by tetrodotoxin, procaine, guanethidine and dibenamine but were not affected by nicotine and phenoxybenzamine. Evoked responses to repetitive stimulation were blocked or suppressed by tetrodotoxin, yohimbine, nicotine, guanethidine and phentolamine. The evoked responses were partially inhibited by phenoxybenzamine. In contrast, evoked responses were enhanced by hexamethonium and dibenamine. These results provide further support for the conclusion that the innervation of the circular smooth muscle of the guinea pig seminal vesicle is mainly alpha-excitatory adrenergic. PMID- 6274059 TI - Lymphoblastoid cell lines, transformed by Epstein-Barr virus, in the enzymatic study of hereditary lysosomal storage diseases. AB - Assay conditions were studied for eight lysosomal enzymes in lymphoblastoid cell lines transformed by Epstein-Barr virus. The transformed lymphoblastoid cells retained all eight enzyme activities, though the levels sometimes differed from those in the peripheral lymphocytes or granulocytes. The levels of these eight lysosomal enzymes were measured in lymphoblastoid cells from 11 patients with hereditary lysosomal storage diseases--GMI-gangliosidosis, a variant of beta galactosidase deficiency (sialidase deficiency with a partial beta-galactosidase deficiency), Tay-Sachs disease, Gaucher disease, Hurler syndrome, Scheie syndrome and I-cell disease--and from 20 of their obligate heterozygotes. No activity of enzymes that were deficient in the respective disease, except I-cell disease, was detected in the lymphoblastoid cells from the patient. In I-cell disease, the cells showed lower levels of some enzyme activities. beta-D-Galactosidase activity from heterozygotes of the patient with GMI-gangliosidosis and alpha-L iduronidase activity from heterozygotes of the patient with Hurler syndrome were in carrier range. On sephadex G-150 gel filtration, beta-D-galactosidase in control material gave two peaks (I and II). In GMI-gangliosidosis, peak II was absent and peak I was markedly diminished. Peak II in the heterozygotes was smaller than that of control. On DEAE cellulose column chromatography of hexosaminidase, two major isoenzymes (hexosaminidase A and B) were detected in control. However, hexosaminidase A was not detected in Tay-Sachs disease, and the ratios of hexosaminidase (Hex) A/Hex B in the parents were lower than those in control. PMID- 6274060 TI - Burkitt's lymphoma in Japan: clinicopathological features of twenty-two patients. AB - The clinicopathological features of 22 Japanese patients with Burkitt's lymphoma were reviewed. There were 14 males and 8 females with a mean age of 9.5 years. Jaw tumors dominated the clinical picture at the time of diagnosis with abdominal tumors second in frequency. The central nervous system and bone marrow were involved during the course of the disease in 32% of the patients. The anti Epstein-Barr (EB) virus antibody titers in Japanese patients were comparable to those found in control group. Serum lactic dehydrogenase (LDH) levels were lower in patients with localized disease than those with advanced tumor. The neoplastic cells were monoclonal B lymphocytes. The patients were very refractory to chemotherapy and had a poor prognosis with a mean survival of 3.8 months from diagnosis. PMID- 6274061 TI - Effect of cadmium on Japanese encephalitis virus infection in mice. 1. Acute and single-dose exposure experiment. AB - When mice were treated with 0.09 mg cadmium chloride (Cd) per mouse once and inoculated i.p. simultaneously with Japanese encephalitis virus (JEV) they showed a significant difference in the incubation time and mortality between the treated and untreated groups in repeated experiments. Cd treatment shortened the incubation time and the mortality increased greater than twice compared with the untreated control. This effect was not observed in the case of intracerebral inoculation of JEV. Effects of Cd on antibody formation in mice were also determined. Animals given a single s.c. dose of Cd were immunized with JE inactivated vaccine once simultaneously. When mice were treated with Cd, they did not show low neutralizing and hemagglutination inhibition activities compared with the control mice. Pretreatment of Cd did not affect any mortality or antibody formation. PMID- 6274062 TI - [Metastasis of malignant tumors of the major salivary glands]. PMID- 6274063 TI - Subcellular mechanisms involving vitamin D. AB - During the past 15 years a vitamin D endocrine system has been demonstrated in which vitamin D produced normally in the skin is activated first by conversion in the liver and subsequently in the kidney to a hormonal form, 1,25-(OH)2D3. The production of the hormonal form of vitamin D3, is regulated, and much has been learned regarding the molecular mechanism of the hydroxylations of vitamin D and regarding the physiologic regulators of the 25-OH-D-1-hydroxylase. Much remains to be learned regarding the mechanism whereby the 1-hydroxylase is modulated. 1,25-(OH)2D3 appears to function in the target organs of bone, intestine, kidney, and elsewhere by a nucleus-mediated process. Receptors for 1,25-(OH)2D3 have been clearly demonstrated and characterized in crude form. How the receptor and ligand interact with the nucleus is not clear, nor are the gene products that result from this interaction known. One product, a calcium binding protein, is known but its role in calcium transport is in debate. Although much has been learned in the last decade and a half, much remains to be learned regarding the molecular mechanisms whereby vitamin D brings about its remarkable changes in mineral metabolism. PMID- 6274064 TI - Immunological studies of tissue proteinases. PMID- 6274065 TI - The cascade of membrane events during development of Dictyostelium discoideum. PMID- 6274066 TI - Modulation of human T-cell differentiation markers by 12-O-tetradecanoylphorbol 13-acetate. AB - The analysis of seven differentiation markers following incubation with the tumor promotor 12-O-tetradecanoylphorbol-13-acetate (TPA) was examined in the human leukemic T-cell line MOLT-3. Significant changes were observed in the activity of the markers terminal deoxynucleotidyl transferase (TdT). spontaneous proliferation and the ability of these cells to bind sheep erythrocytes. Levels of human thymus-leukemia-associated antigen (HThy-L) recently identified as a low molecular weight form of adenosine deaminase (ADA), were reduced by about 50%. No significant changes were observed in ecto-5'-nucleotidase [5'-NT) activities, in the proliferative response to PHA, or in the expression of IA-like antigens. These data and the time kinetics of the changes suggest that following incubation of these T-lymphoblasts with TPA there is a sequential loss of TdT, loss of the capacity for spontaneous proliferation, and the appearance of receptors for sheep erythrocytes. Subsequently there is a decrease in the level of HThy-L/ADA. This sequence appears to follow that proposed for prethymic precursor T-cell differentiation following activation with thymic epithelium. PMID- 6274067 TI - Preparation of varicella-herpes zoster immunoglobulin. AB - The efficacy of zoster immunoglobulin in prophylaxis of varicella and herpes zoster is reported by many authors. The screening for zoster's antibodies is performed in the blood donor population and in persons convalescing from herpes zoster. The results show that 25 per cent of blood donors and 83.5 per cent of convalescents, respectively, have CF antibody titer of l6. Varicella zoster immunoglobulin prepared by alcohol fractionation has a fluorescent antibody titer of 1,024 and a CF antibody titer of 512. These results are discussed and compared with those of the literature. PMID- 6274068 TI - Histological typing of lung tumours. AB - The WHO Histological classification of Lung Tumours, published in 1967, has been revised. The main features are as follows: Squamous cell carcinoma (epidermoid carcinoma) has the same definition as in the original version, i.e., the identification of keratin and/or intercellular bridges by light microscopy. Three degrees of histological differentiation are described. Dysplasia and carcinoma in situ are discussed. Small cell carcinoma is divided into oat-cell carcinoma, an intermediate cell type and a category for oat-cell carcinomas combined with other major types. Adenocarcinoma includes the acinar, papillary and bronchiolo alveolar forms and the solid carcinomas with mucus formation (previously part of the large cell carcinoma group). Mesothelial tumours are divided into fibrous, epithelial and biphasic subtypes. A number of less common tumours and tumour-like lesions are defined. PMID- 6274069 TI - Correlation between the presence of natural antitumor antibodies and activation of MuLV endogenous virus in BALB/c mice. AB - Individual 3-month-old or 12-month-old BALB/c mice, as well as 5-month-old nu/nu or nu/ + BALB/c mice, showed a direct correlation between the serum level of natural antitumor cytotoxic antibodies and the capacity of spleen cells to infect SC-1 cells permissive for murine ecotropic viruses. Pooled or individual sera from 3-month-old BALB/c mice, negative for the presence of natural antitumor cytotoxic antibodies and whose spleen cells were unable to infect the SC-1 cells, were negative both for SC-1 cells and SC-1 cells infected by MuLV. On the contrary, pooled or individual sera from 15-month-old BALB/c mice, positive for the presence of natural antitumor antibodies and with infecting spleen cells, were cytotoxic for infected SC-1 cells but not for the uninfected ones. The infection of SC-1 cells by MuLV could be inhibited by 3-month-old spleen cells, and this effect was suppressed by depriving the inhibiting spleen cells of T cells by means of an anti-Thy-1 antibody plus complement. The cells with infectious capacity did not belong to the T-cell compartment, as demonstrated by the lack of infection after passing the infecting spleen cells through an anti-Ig column, whereas T-deprivation did not modify the infectious capacity. A natural anti-gp70 monoclonal antibody, which exerted a complement-dependent cytotoxic effect on tumor cells, strongly inhibited the infection of the permissive SC-1 cells by MuLV. PMID- 6274071 TI - Some aspects of pituitary function after neuroadenolysis in patients with metastatic cancer. AB - The effects of neuroadenolysis on plasma titres of beta-endorphin, beta lipotropin, ACTH, TSH and prolactin have been investigated in five patients with metastatic cancer who responded to the treatment and have been in remission for more than four years and in five others who were undergoing the treatment for the first time for pain due to cancer metastases. beta-Endorphin, beta-lipotropin and ACTH titres were within the normal ranges of values in both categories of patients but post-neuroadenolysis titres of these peptides were higher than those before the treatment. The ability to secrete TSH and prolactin and to respond to thyroid stimulating hormone releasing hormone (TRH) remains intact following the treatment. However, whereas basal TSH titres and response to TRH was lower in the majority of patients, no such effect was observed on prolactin secretion. Plasma titres of prolactin and TSH were below the sensitivity of the method in the five patients who are in remission for more than four years. These preliminary findings suggest that neuroadenolysis probably affects some mechanism(s) associated with the control of beta-endorphin, beta-lipotropin and ACTH synthesis. PMID- 6274070 TI - A two-stage immunocytochemical method for oestrogen receptor analysis: correlation with morphological parameters of breast carcinomas. AB - An immunocytochemical method for oestrogen receptor (ER) analysis of breast carcinomas is described and compared with the dextran-coated charcoal (DCC) assay and a cytochemical method. The method described is superior to the purely cytochemical, especially in terms of sensitivity, demonstration of nuclear receptors and correlation with the DCC assay. The DCC assay is reliable except in the case of poorly cellular tumours: in these the immunocytochemical method is far superior, being independent of tumor cellularity. With this new method, lobular carcinomas are shown to be ER-rich compared with their ductal counterparts, a conclusion currently disputed, largely because methods employed mostly fail to take account of tumour cellularity. A combination of the DCC assay and the immunocytochemical method here described gives the maximum information about the ER-status of a particular carcinoma. PMID- 6274072 TI - Sarcomatous variant of Wilms' tumor. A light and immunohistochemical study of four cases. AB - Four cases of infantile sarcomatous renal tumors are reported. Rhabdomyoblastic differentiation was investigated in these tumors by the immunoperoxidase method using antimyoglobin sera. Rhabdomyoblastic cells were found in 2 cases and in a metastasis of one case in which the primary tumor was composed by undifferentiated cells negative for myoglobin stain. The histogenesis of these neoplasias is discussed, and it is suggested that they are a sarcomatous variant of Wilms' tumor. PMID- 6274073 TI - [Carcinoma of the pancreas presenting with urological symptoms. Report of two cases (author's transl)]. AB - Description of two cases of pancreatic cancer with first symptoms in the urological field. A report is given about symptoms and progress in these two cases. Suggestions for treatment are made. PMID- 6274074 TI - Use of 99mTc-DMSA in renal cortical abscess. PMID- 6274075 TI - [State of the acoustic analyzer in patients with neoplasms of the pharynx and parapharyngeal space]. PMID- 6274076 TI - [Laryngeal histiocytoma]. PMID- 6274077 TI - [Current concepts of the role of the kallikrein-kinin system in the pathogenesis of acute arterial obstruction and its sequelae (a review of the Soviet and foreign literature)]. PMID- 6274078 TI - [Recanalization of the hepatic ducts in high obstructive blockage]. AB - Forty three patients were subjected to 48 operations for restoration of the passability of lobular ducts when the latter were obturated by tumor (36 patients) and by alveococcus (7 patients). Ten patients died immediately after operation. Death was caused by acute hepato-renal insufficiency and acute pancreatitis. Duration of life after operation was from 7 months to 3 years. Best results were obtained in patients with alveococcus of the portal fissure. The authors make a conclusion that recanalization of main biliary ducts in obturation ileus is less traumatic as compared with other operations, improves the general state of the patient and prolongs the duration of their life. PMID- 6274079 TI - [Comparative evaluation of the diagnostic importance of determining 5 nucleotidase and alkaline phosphatase in the blood serum in liver and biliary tract diseases]. AB - Under examination were 186 patients with diseases of the liver and biliary ducts. In patients with mechanical jaundice resulting from cholelythic disease high indices of the activity of 5-nucleotidase and alkaline phosphatase were revealed in 80--85% of cases, in patients with carcinoma of the head of the pancreas--in 100%. The investigation of these tests and blood serum transaminases in dynamics is of great importance for differential diagnosis of diseases of the liver and biliary ducts manifesting themselves by the jaundice syndrome. PMID- 6274080 TI - [Influenza epizootic in swine due to strain A (Victoria/3/75 H3N2)]. AB - Studies were carried out on the first grippe epizootic in pigs in Bulgaria. The affected animals showed a rise in temperature (40.5 to 41.8 degrees C), loss of appetite, abdominal type of respiration, and noisy and painful cough. The infection ran its course with a picture typical of a virus disease which did not respond to broad-spectrum antibiotic treatment. On an average the duration of the clinical course lasted 4 to 5 days, affecting almost all swine on a pig breeding farm, with single death cases. The morphologic changes observed in animals killed for diagnostic purposes were typical of a grippe infection. Isolated was a grippe virus A (Antimovo) 1/80/H3N2) the preliminary typisation of which showed close relation to strain A (victoria) 3/75/H3N2/. Serologic investigations using double blood samples from the affected pigs in double immunodiffusion test and hemagglutination-inhibition test demonstrated the etiologic role played by the isolated grippe virus. PMID- 6274081 TI - [Comparative pathomorphological studies of piglets infected with different strains of the pseudorabies virus]. AB - Studied were morphologically the central nervous system and the internal organs of a total of 40 experimentally infected pigs-10 infected with the virulent strain B (5.2 C 10(6) PFU), 10 infected with the slightly virulent strain K (4.8 X 10(6) PFU), and 20 infected with the vaccinal mutant strain MK 25 (2.4 X 10(6) PFU), along with 4 control pigs. Histologically, all treated animals showed changes in the central nervous system characteristic of nonsuppurative meningoencephalitis. The lesions were more strongly expressed in the pigs infected with the virulent strain. Those of the animals treated with the MK strain presented changes that were similar to the changes observed in the pigs of the remaining two groups. By intensity and size, however, they were more slightly manifested and tended toward rapid limitation and lower frequence and dimension. The animals infected with strain K mad also changes in the lungs - interstitial pneumonia of lymphohistiocytis type. Those injected with the MK 25 strain presented numerous macrophages in the tonsils and lung lymph nodes, which was referred to as the morphologic expression of immunogenesis. The morphologic differences observed in the course of Aujeszky's disease in the individual test groups of pigs were due to the varying virulence and tissue tropism of the pseudorabic virus strains used. Studies revealed that the strongly virulent strains were predominantly neurotropic, while the slightly virulent ones were to a lesser extent neurotropic but were shown to be strongly pneumotropic. PMID- 6274082 TI - [Inactivated vaccines against rhinopneumonitis in horses]. AB - Attempts were made to produce inactivated vaccines against horse Herpes virus 1, using various inactivating agents and adjuvants, Best results were obtained with vaccine No 3 (glutaraldehide inactivator and "CTC" adjuvant). Used were two strains of the virus (St. Karaja and Varna). isolated in this country in cell cultures of a sucking pig kidney. Vaccine No 3 showed good immunogenic properties. Its application resulted in the full cease of abortions and respiratory diseases on the base of infection with the horse Herpes virus 1. The vaccination protects newborn colts from rhinopneumonitis if reimmunization of mares is carried out in the 6th-7th month of pregnancy. PMID- 6274083 TI - [Isolation of an adequate antigen for the diagnosis of enzootic leukemia in cattle with the agar gel immunodiffusion test]. AB - An effective antigen was obtained, presenting a leukosis virus. The culture liquid, and occasionally the cells of the FIK cell line permanently infected with the agent of bovine leukosis served as a source of virus. The antigen, possessing a very good activity was produced by means of precipitation of the virus with 30 per cent ammonium sulfate, dialysis, and additional, concentration with polyethylene glycol-4000 until a 100-fold concentrations as against the initial volume of the virus was attained. Diluted antigen at 1:1 and 1:5 and leukosis serum produced in agar gel a precipitation line for 24 to 48 hours. In terms of activity and specificity this line did not differ from the line formed by glucoprotein antigens obtained from West Germany and the United States. With a normal bovine serum and sera containing Rota, and Parainfluenza-3 antibodies no precipitation line was formed. The good properties of the antigen were confirmed by joint investigations of bovine sera in this country and the Soviet Union with identical results. A total of 26 000 sera were examined with the newly obtained full value antigen, establishing the spread of the leukosis infection in this country as well as the fact that it had been imported from abroad. PMID- 6274084 TI - Host cell reactivation of uv- and X-ray-damaged herpes simplex virus by Epstein Barr virus (EBV)-transformed lymphoblastoid cell lines. PMID- 6274085 TI - Structural homology of the major internal proteins of endogeneous type C viruses of two distantly related species of Old World monkeys: Macaca arctoides and Colobus polykomos. PMID- 6274086 TI - Hematopoietic cells transformed in vitro by REVT avian reticuloendotheliosis virus express characteristics of very immature lymphoid cells. PMID- 6274087 TI - Revertants of rats cells transformed by avian erythroblastosis virus. AB - Morphological revertants of the avian erythroblastosis virus (AEV)-transformed rat cell line ATla were isolated and characterised. The revertants are similar to the uninfected parental rat cell line in that they have regained an organized cytoskeleton and they are no longer capable of anchorage-independent growth. The pattern of integrated viral DNA in the revertants is indistinguishable from that of the transformed parent. However, the revertants do not express the integrated viral genome at either the mRNA or protein level. Phenotypic reversion thus is probably .due to reduced transcription of the AEV-transforming gene below a threshold necessary to induce morphological transformation. PMID- 6274088 TI - The HN glycoprotein of Sendai virus: analysis of site(s) involved in hemagglutinating and neuraminidase activities. PMID- 6274089 TI - Structural changes in the DNA of Marek's disease virus during serial passage in cultured cells. PMID- 6274090 TI - Induction of Epstein-Barr virus early antigens by intercalating chemicals in B95 8 cells. PMID- 6274091 TI - Epstein-Barr virus-induced deoxynuclease and the reutilization of host-cell DNA degradation products in viral DNA replication. PMID- 6274092 TI - Identification of a 20,000-dalton protein in SSV-transformed nonproducer cells. PMID- 6274093 TI - Epstein-Barr virus-induced early polypeptides in Raji and NC37 cells activated by diterpene ester TPA in combination with N-butyrate. PMID- 6274094 TI - Extrachromosomal bovine papillomavirus type 1 DNA in hamster fibromas and fibrosarcomas. PMID- 6274095 TI - [Crystalline inclusion in hepatocytes in cutaneous pophyria tarda]. PMID- 6274097 TI - [Disorders of neuromuscular apparatus in children with rheumatoid arthritis (electrophysiological data)]. PMID- 6274096 TI - [Cyclic adenosine monophosphate in the fatty tissue of breast and lung cancer patients]. AB - Content of cAMP was studied in fatty tissue of 33 patients with tumor of mammary gland, of 27 patients with tumor of lung tissue and of 27 persons (15 men and 12 women), which constituted the control group. A procedure involving competitive binding of the nucleotide with protein was used. The data obtained were related to the size and amount of adipocytes in fatty tissue. The size of adipocytes was distinctly larger and content of cAMP in fatty tissue was slightly lower in patients with tumor of mammary gland as compared with the corresponding controls. With ageing content of fatty tissue cAMP tended to increase in men and to decrease in women. Negative correlation between the content of cAMP in fatty tissue (pM per kg of body mass) and the size of adipocytes, characteristic for the control group, was not found in the patients with the both forms of tumor. Significance of impairments in synthesis of cAMP in adipocytes for development of specific metabolic patterns of adipose tissue in oncological patients is discussed. PMID- 6274098 TI - [Nicotinamide as an effective agent against endogenous hypocorticism during prolonged corticosteroid therapy]. PMID- 6274099 TI - [Nitrate content in vegetables]. PMID- 6274100 TI - A dynamic investigation in antibodies to influenza and parainfluenza viruses in patients suffering from chronic bronchitis and bronchial asthma. PMID- 6274101 TI - [Autoantibodies against beta-adrenergic receptors (author's transl)]. PMID- 6274102 TI - [IgM antibodies against cytomegalovirus in pregnancy and incidence of intrauterine infections in pregnant women with positive IgM antibody reaction (author's transl)]. AB - In 47 of 620 pregnant women (7.6%) examined during the first, second and third trimester, it was possible to identify IgM, antibodies against late antigen (LA) of the cytomegalovirus during one or several examinations. 6 of 37 examined children of mothers with positive IgM antibody reaction were found to have intrauterine infection, although there had been no clinical signs of an infection at the time of delivery. All 6 children with intrauterine infection were from 32 mothers (18.8%) with IgM antibodies during the first trimesters. The IgM antibodies of the mothers with children having intrauterine infection did not differ fundamentally from the IgM antibody levels of the mothers with non infected children with regard to titration value and persistency. PMID- 6274103 TI - [Fungal infection of the scalp by Trichophyton soudanense treated with ketoconazole (author's transl)]. AB - Case of 25 years old black Saracole female presenting tinea capitis caused by Trichophyton Soudanense. This patient lives in Senegal. She was 5 years old on the onset of the disease, which has evolved continuously for 18 years. Systemic administration of Ketoconazole, 400 mg a day during 25 days, had totally cured the patient clinically and mycologically. PMID- 6274104 TI - [Psychometric aspects of estrogen therapy in climacteric woman (study of 122 women after menopause]. PMID- 6274105 TI - [Ultrastructure of detritus synovitis ("hydroxylapatite synovitis?") (author's transl)]. AB - Detritus synovitis is a sequel of cartilage and osseous destruction in diarthrosis. Cartilage and bone fragments are enclosed by granulation tissue. Remnants of the destroyed bone and calcified cartilage appear as defined accumulations of hydroxylapatite crystals in the granulation tissue. These hydroxylapatite tissues resemble those considered by other authors as being responsible for a "third crystal synovitis, namely, hydroxylapatite synovitis". However, the findings communicated here allow us to conclude that these crystals are the sequel of a destruction of diarthrosis, so that it is not justified to consider hydroxylapatite synovitis as a separate disease entity. PMID- 6274106 TI - Ultrasound: first mandatory step in diagnosis of abdominal masses in children. PMID- 6274107 TI - [Mesoblastic nephroma--a malignant tumor with low grade of malignancy]. AB - The mesodermally differentiated and only in newborns and young infants occurring mesoblastic nephroma is regarded as benign. Recidivations with malignant course, inclination to infiltration and richness in mitosis make arise doubts about the benignity despite good prognosis. The results of a proliferation-kinetic examination received by an autoradiographic in vitro method of a mesoblastic nephroma which was observed in a 20-day-old infant are characteristic for a malignant tumour of low degree of malignity: 3H-thymidine marking index 5.8%, DNA synthesis time 18.2 hours, mean generation time 141.1 hours (= 5.9 days). Growth fraction 0.28 and potential doubling time of the tumour 16.4 days. These data decisively deviate from those of the Wilms-tumours, which are cell-kinetically characterized as extraordinarily quickly growing, highly malignant tumours. The position of the mesoblastic nephroma in the system of the dysontogenetic renal tumours is briefly discussed. The ureteronephrectomy performed in time is the only necessary therapeutic measure, since also the mesoblastic nephroma follows the general rule of dysontogenetic tumours that the prognosis is the better the younger the infant is. PMID- 6274108 TI - [The latex-IgM-test in virus diagnostic: determination of cytomegalovirus antibodies (author's transl)]. AB - We examined the practical value of the Latex-IgM-test in diagnostic virology. Sera of patients suspected to have a cytomegalic infection were used for serological investigation. Cytomegalovirus produced in cell cultures was incubated with serum from patients. The resulting virus-antibody complex was removed by centrifugation and sensibilized Latex particles were added (anti IgG and anti IgM). The quantity and type of the bound antibody could be determined in this way. The sera were checked by parallel determination by another reliable method, the neutralization test. Immunoglobulins G and M separated and tested for antibodies. The usefulness of the Latex-IgM test for the routine diagnostics could be evaluated in the present study. PMID- 6274109 TI - [Analysis and improvement of culture processes of pathogenic microorganisms]. PMID- 6274110 TI - [Histochemical and electron-microscopic study of brain structures during the acute period of experimental craniocerebral injuries]. AB - Three stages of cerebral metabolism differing in tension were revealed in the acute period of closed craniocerebral trauma. In the mobilization stage (1 hour after trauma) enzymatic reactions reflecting the activity of aerobic and anaerobic glycolysis are sharply intensified. In the resistance stage (from 3 hours to 2 weeks after trauma) stable conversion to anaerobic glycolysis is noted, which leads to marked deficiency in macroergic compounds. Activation of alpha-glycerophosphate, pentose shunt by means of pharmacological agents in combination with hypothermia may be recommended in this stage. In the restoration stage (more than 2 weeks after the trauma) the metabolic processes are gradually normalized and therapy need not be so intensive. PMID- 6274111 TI - On the mechanism of red blood cell shape change and release of spectrin-free vesicles. AB - Changes in the negative change densities on the outside (by raising the phosphatidic acid concentration) or on the inside of the red blood cell membrane (by a breakdown of the polyphosphoinositides and a concomitant diacylglycerol production) do not explain the last step in echinocytosis, the release of spectrin-free vesicles. PMID- 6274112 TI - Ultrastructural modifications of the erythrocyte membrane in Down's syndrome. AB - Spin label studies of erythrocyte membranes from patients with Down's syndrome showed no differences in the rigidity of membrane lipids with respect to normal subjects and a tendency for alterations in the state of membrane proteins. Electron microscopic studies demonstrated the occurrence of ultrastructural defects in membranes from trisomics which can be due to accelerated red cell aging as similar alterations were found in the fraction of oldest (most dense) cells in normal subjects. PMID- 6274113 TI - Molecular cloning of DNA sequences coding for mouse embryonic globins. AB - Yolk sac derived erythroid cells in mouse embryos synthesize four embryonic globins of which two are alpha-like and two are beta-like. Pure globin messenger RNAs from these cells were used as templates for two successive polymerizing reactions and a mixture of double stranded cDNAs coding for the four globins was obtained. These molecules were blunt-end ligated to an ECoR1 digested pBR322 plasmid and the recombinant plasmids were used to transform E. coli Hb101. Bacterial clones which proved positive upon hybridization with 32P-labelled embryonic globin cDNA were amplified and their plasmid DNA was isolated. Three different plasmids were studied, namely no. 2, 16 and 54. The restriction map of these plasmids showed that: 1) plasmid no. 2 and 54 had lost extensive DNA sequences comprising the genes responsible for tetracycline resistance; 2) the size of inserted sequences ranges from 427 base pairs of plasmid no. 16 to about 280 base pairs of plasmid no. 54; 3) plasmid no. 2 does not share any of the studied restriction sites with the other plasmids, while no. 2 and 54 have at least one site in common. The coding properties of inserted DNA were determined by positive hybrid translation showing that no. 2 codes for the alpha-like embryonic chain x, while no. 16 and 54 code for a beta-like embryonic chain, either y or z. PMID- 6274114 TI - Studies on pyruvate kinase deficiency, pyrimidine 5-'nucleotidase deficiency and adenosine deaminase overproduction. AB - Using recently established ICSH recommended methods, red cell pyruvate kinases (PK) of 20 patients with PK deficiency were characterized and 7 new PK variants were found. Analysis of partially purified red cell pyrimidine 5'-nucleotidase (P5N) from a patient with P5N deficiency provided the evidence for a structural alteration of the enzyme protein. Red cell adenosine deaminase (ADA) from a patient with 40-fold increase in ADA activity associated with hemolytic anemia was purified and compared with that from normal subjects. It is most conceivable that the increased ADA activity represents increased amount of structurally normal enzyme. PMID- 6274115 TI - Changes in the enzymes related to 2,3-bisphosphoglycerate metabolism in developing erythroid cells. AB - Measurements of glycerate-2,3-P2 and hemoglobin in the developing erythroid cells indicated that the glycerate-2,3-P2 level rose during erythroid differentiation in a linear relationship to the hemoglobin level, suggesting the presence of regulation to accumulate both substances synchronously. The accumulation of glycerate-2,3-P2 was found to be primarily attributable to the increase in glycerate-2,3-P2 synthase activity. The activities of phosphofructokinase and pyruvate kinase changed so as to be favourable for glycerate-2,3-P2 accumulation. The increase in glycerate-2,3-P2 synthase activity was shown to be caused by an increase in the enzyme protein. Synthesis of glycerate-2,3-P2 synthase protein was proved in bone marrow erythroid cells and in reticulocytes. PMID- 6274116 TI - Antagonistic effect of cAMP and cGMP on the kinetic behaviour of phosphofructokinase from rat erythrocytes and reticulocytes. AB - Fructose-6-phosphate (F6P)-saturation curves (up to 5 mM F6P) for phosphofructokinase (PFK) have been studied at physiological pH (7.1) and inhibitory (1.5 mM) or non-inhibitory (0.25 mM) ATP levels, in rat erythrocytes and reticulocytes. The addition of 300 microM cAMP to control samples activates the enzyme and displaces F6P-saturation curve towards the left, while the addition of cGMP inhibits the enzyme and shifts the curve to the right. The cAMP positive allosteric effect is more evident at inhibitory ATP levels, while the inhibitory effect of cGMP is very similar at both ATP levels. This antagonistic effect is exerted at the same regulatory site, since cAMP also activates the enzyme when cGMP is previously present in the reaction mixture. The physiological significance of this antagonism is not yet clear. PMID- 6274118 TI - Medical treatment of insulinoma. PMID- 6274117 TI - Muscimol, a psychoactive constituent of Amanita muscaria, as a medicinal chemical model structure. PMID- 6274119 TI - [The study of the hepatic clearance of 131I-rose-bengal in epileptic patients treated with phenobarbital associated or not with hydantoin (author's transl)]. PMID- 6274120 TI - Intranuclear inclusions in Bence Jones kappa myeloma. PMID- 6274121 TI - Two cases of hereditary hemolytic anemia with pyrimidine 5'-nucleotidase deficiency. PMID- 6274122 TI - [Cytochemical studies of peripheral blood granulocytes of workers with occupational exposure to lead compounds]. AB - In 50 workers of a non-iron metallurgic plant exposed in their work to lead and trace amounts of cadmium and zinc cytochemical reactions were carried out in peripheral blood granulocytes. The control group comprised 30 men. In the investigations the degree of intoxication with lead compounds and the effect of the length of exposure ion the cytochemical reactions were taken into account. It was observed in these investigations that chronic exposure to lead and trace amounts of the remaining elements caused changes in the activity of acid phosphatase, alkaline phosphatase, lactic dehydrogenase and MOP. In particular, a significant fall of MOP and acid phosphatase activity and increased LDH activity in subjects exposed to lead with biochemical evidence of lead absorption deserve attention. With longer exposure to lead the activity of MPO decreased in workers with evidence of lead absorption. PMID- 6274123 TI - [Case of stomach cancer with erythroleukemic reaction and thrombopenic purpura]. PMID- 6274124 TI - Antibodies in oligoclonal immunoglobulins in CSF from patients with acute cerebrovascular disease. AB - Thin-layer polyacrylamide gel isoelectric focusing (PAG IEF) of CSF and serum, and subsequent immunofixation with viral and structural brain components followed by autoradiography revealed in eight out of nine selected patients with oligoclonal CSF IgG and cerebrovascular disease local synthesis within the CNS of antibodies against one or more of the viruses tested: six patients against measles, five against herpes simplex virus type 1, and two against varicella virus. This finding may reflect a polyclonal B cell activation secondary to brain damage and elaboration of certain structural brain components. None of the patients had local synthesis of antibodies against the other viruses tested (mumps, rubella and cytomegalovirus), or against structural brain components (crude saline, lipid-proteolipid, myelin basic protein extracts from human brain and purified bovine myelin basic protein). PMID- 6274125 TI - The pathology of allyl chloride neurotoxicity in mice. AB - Allyl chloride is known to produce a neuropathy in man after occupational exposure to its vapour. The present study describes the neuropathy which develops in mice given allyl chloride by mouth. Mice were dosed three times weekly with 300 or 500 mg/kg allyl chloride for periods from 2-17 weeks. Functional disability was observed in some animals. Apart from evidence of focal kidney damage in 70% of dosed mice, pathological changes were restricted to the nervous system. Nerve fibre degeneration was found in many peripheral nerves and in roots, tending to be more marked distally and to affect more motor than sensory nerves. Degenerated fibres were also found in dorsal, ventral and lateral columns of the spinal cord. Males were more severely affected than females. Increased numbers of filaments were on early axonal change, occurring multifocally and apparently preceding axonal degeneration. No neuronal death was observed, but occasional anterior horn and dorsal root ganglion cells showed some morphological changes. Vacuolated lesions mainly due to swelling of astrocytes and their processes were found in the ventral horn in cervical and lumbar regions of spinal cord. Animals appeared to become tolerant to allyl chloride after continuous dosing. This neuropathy appears to be a central-peripheral distal type of axonopathy. PMID- 6274126 TI - A new type of non-progressive sensory neuropathy in children with atypical dysautonomia. AB - Three cases of non-progressive, sensory neuropathy with dysautonomia are presented. Light and electron microscopy on whole sural nerve biopsies revealed an almost total lack of myelinated nerve fibres. The total fibre count was also reduced as was the total number of Schwann cell nuclei. No degenerative phenomena were seen within the nerve fibres. The aberrations are probably caused by a maldevelopment of the neural crest implying a stunted proliferation and growth of sensory and autonomous neurones as well as a reduced proliferation of Schwann cells. Since the morphology and clinical features differ from that in other cases of sensory neuropathy with dysautonomia the three present cases are considered to represent a new type of the disease. PMID- 6274127 TI - Galactose neuropathy. Permeability studies, mechanism of edema, and mast cell abnormalities. AB - Galactose neuropathy is characterized by progressive endoneurial edema manifested by a gradual increase in endoneurial fluid pressure. Edema accumulates via a unique mechanism of osmotic force generated by products of the polyol pathway, synthesized within the endoneurial compartment. This paper presents morphologic findings showing firstly, that blood nerve barrier permeability to horseradish peroxidase complexes appears unchanged and secondly, peripheral nerve edema in this condition is restricted to extraganglionic endoneurium sparing the spinal ganglia and adjacent roots. Thirdly, mast cells accumulated in significant numbers and electron microscopy revealed degranulation. There was no evidence of edema in Schwann cell cytoplasm, the putative site of galactitol accumulation via the sorbitol pathway. These findings are discussed with respect to diabetic neuropathy for which galactose intoxication is a useful experimental model. PMID- 6274128 TI - Treatment of Bell's Palsy. An analysis of the available studies. AB - In this paper the basic requirements for a clinical therapeutic trial in Bell's palsy are discussed. The available therapeutic studies are analysed in view of these requirements. The following conclusions seem justified: (1) Galvanic stimulation has not been proved to lead to a better recovery. (2) There is no evidence that decompression of the mastoid part of the facial nerve in the second or third week has a positive effect. Whether surgery at an earlier stage or decompression of the complete intratemporal part of the nerve is effective, has not yet been studied. (3) There is no convincing proof that prednisone administration up to a daily dose of 60-80 mg during the first days has any effect. PMID- 6274129 TI - Studies on the intracellular cAMP metabolism of tonsillar lymphocytes. AB - cAMP metabolism, including the intracellular cAMP level and adenylcyclase activity, was studied in tonsillar and peripheral lymphocytes. The basal activity of adenylcyclase was significantly higher in tonsillar lymphocytes than in peripheral lymphocytes, although the basal level of intracellular cAMP did not differ. The responsiveness of the intracellular cAMP level and adenylcyclase activity stimulated by 10(-5) M isoproterenol, or 10(-5) M prostaglandin E1, which was thought to stimulate this enzyme through a specific receptor, was significantly lower in tonsillar lymphocytes than in peripheral lymphocytes. Moreover, the responsiveness of adenylcyclase activity to 10(-2) M NaF, which was thought to stimulate this enzyme directly, was also significantly lower in tonsillar lymphocytes. Tonsillar T lymphocytes represented a lower cAMP basal level than tonsillar B lymphocytes, but no difference was observed in the responsiveness of intracellular cAMP to the above agents. Tonsillar lymphocytes derived from patients with recurrent tonsillitis represented significantly lower cAMP responsiveness than those from patients of other chronic tonsillitis. Furthermore, in the tonsillar lymphocytes from patients with a recent attack of acute tonsillitis, a more depressed responsiveness of the intracellular cAMP level to the above agents was recognized. The cAMP basal level was not changed by these pathological alterations of tonsillitis. PMID- 6274130 TI - Brain cortex gangliosides and (Na+, K+)ATPase system of the stria vascularis in guinea pig. AB - This paper presents the results of an investigation into the possibility that gangliosides (brain cortex glycosphingolipids) are capable of being functionally incorporated into cell membranes and of interfering with enzymatic activity of the ATPase system on the stria vascularis and spiral ligament in the guinea pig. Labelled gangliosides (3H-GM1) were incorporated into the cell membranes of the tissues under examination. (Na+, K+)ATPase activity increased l0 minutes after intravenous injection of gangliosides and prevented the decrease in (Na+, K+)ATPase system produced by ethacrynic acid. The action of gangliosides on the (Na+, K+)ATPase system is discussed. PMID- 6274131 TI - Computerized documentation in middle ear surgery. Methods and clinical experiences. AB - A computerized system for recording results of ear surgery is presented. Specially designed forms are filled in at the time of operation as well as 2 months, 6 months and annually for 1-5 years after operation. The data are processed in a Univac 1100-80 by specific programmes written in FORTRAN. A Follow up programme gives up-to-date information on hearing and healing after different types of operations. An Annual Production Control programme gives more detailed information about various correlations, such as results in draining ears, frequency of residual cholesteatoma etc. Four years experience with the system and obtainable data is discussed. PMID- 6274132 TI - Acute otitis media. A prospective clinical study of myringotomy and antimicrobial treatment. PMID- 6274133 TI - [Juvenile nasopharyngeal angiofibroma]. PMID- 6274134 TI - [The manner of technetium uptake in salivary tumors and its implications in diagnosis]. PMID- 6274135 TI - Familial Fanconi syndrome with malabsorption and galactose intolerance, normal kinase and transferase activity. A report on two siblings. AB - Two siblings of Turkish-Assyrian extraction, whose parents were first cousins, had poor appetite, slow weight gain and retarded psychomotor development. When given milk the galactose concentration in blood increased. An oral galactose load showed a markedly reduced capacity to metabolize galactose. Fanconi syndrome was present as in classical galactosemia. A galactose-free diet reduced the aminoaciduria but did not normalize the renal tubular function nor the children's general condition. Galactokinase and galactose-1-phosphate uridyltransferase activities in red blood cells were normal. The physical appearance of the children (sparse subcutaneous fat, thin extremities and distended abdomen) and the results of vitamin A and xylose absorption tests, were in accordance with a malabsorption condition. Glucose, however, seemed to be absorbed normally from the gut. There was no evidence of primary liver disease. Since the condition did not normalize with a galactose-free diet, an enzyme defect of galactose metabolism is unlikely. Instead, a more general transport defect with autosomal recessive inheritance is proposed. PMID- 6274136 TI - Fine structure of cystosarcoma phyllodes with reference to smooth muscle tumors. AB - The fine structure of the stomal component of cystosarcoma phyllodes was compared with that of intracanalicular fibroademona, leiomyoma and leiomyosarcoma. The stromal cells of fibroadenoma was intermediate between fibroblasts and smooth muscle cells and may be designated as myofibroblasts. The stromal cells of cystosarcoma phyllodes have fulfilled the ultrastructural criteria for smooth muscle cells. Therefore, cystosarcoma phyllodes should be regarded as smooth muscle tumors or those of myoepithelial origin. PMID- 6274137 TI - Malignant fibrous histiocytoma of bone associated with focal hemangiopericytomatous pattern. AB - A case of malignant fibrous histiocytoma of the tibia of a 49-year-old Japanese farmer was presented. The patient was diagnosed as malignant hemangiopericytoma by biopsy, whereas the amputated material showed a marked pleomorphism representing features of malignant fibrous histiocytoma. Ultrastructurally, undifferentiated mesenchymal cells, intermediate cells between the undifferentiated mesenchymal cells and fibroblastic cells, fibroblastic cells and histiocytoid cells including bizarre giant cells were confirmed in a broad modulating spectrum. The hemangiopericytomatous lesion was predominated by intermediate cells, the ultrastructure of which bore a close resemblance to that of pericytes, and was regarded to be resulted from a differentiation of undifferentiated mesenchymal cells towards the pericyte. PMID- 6274138 TI - Progressive multifocal leukoencephalopathy. Neuropathology and virus isolation. AB - Mechanism of demyelination in progressive multifocal leukoencephalopathy (PML) was elucidated by demonstrating ultrastructurally the relationship between the affected glial cells and myelin sheaths. The myelinating cells in the central nervous system, namely oligodendroglias, were specifically attacked by PML-virus with eventual primary destruction of myelin sheaths. An attempt to isolate PML virus from the autopsy material was made. The presence of virus particles in the extracts from the brain, reactions of virions with antibodies and evidence of virus proliferation in the primary human fetal glial cell culture were shown by various techniques including negative staining, immunofluorescence, immunoelectron microscopy, and electron micrography. The isolated virus was antigenically identical with JC type of papovaviruses. PMID- 6274139 TI - Interaction of feline sarcoma virus (FeSV) and mycoplasma. AB - A factor present in the supernatant of an established human glioma cell line U 251 MG strongly suppresses feline sarcoma virus (FeSV) focus forming activity on feline embryo fibroblasts. The factor was identified as mycoplasma arginini. The enriched mycoplasma fraction had no cytpathogenic effect on the glioma cells or on the embryonic feline indicator cells. An antiserum prepared against this strain of mycoplasma abolished the inhibition. The exact mechanism is not known but arginine depletion in the medium seems to be an important factor. PMID- 6274140 TI - Serum prolactin levels and immunohistochemical localization of prolactin in trophoblastic disease. AB - The circulating levels of chorionic gonadotropin, pregnancy specific beta-I glycoprotein, prolactin and oestradiol were estimated by radioimmunoassay in 219 serum samples from 18 patients with choriocarcinoma and 45 samples from 8 patients with hydatidiform mole. Prolactin and oestradiol concentrations were higher in samples which also contained trophoblastic markers. This difference was also seen in patients with choriocarcinoma and it was not due to chemotherapy, since the difference between hCG positive and hCG negative samples was maintained during chemotherapy. Prolactin was found by immunoperoxidase technique in the syncytiotrophoblast of normal placenta throughout gestation, and also in hydatidiform mole, invasive mole and choriocarcinoma. It is suggested that the malignant syncytiotrophoblast may contribute to circulating prolactin levels either by direct secretion of prolactin or by oestrogen stimulation of the pituitary. PMID- 6274142 TI - Susceptibility of continuous lines of monkey kidney cells to influenza and parainfluenza viruses in the presence of trypsin. AB - LLC-MK2, GMK AH-1, BSC-1, and Vero cells were compared in titrations of recent isolates and laboratory strains of influenza A and B and parainfluenza types 1, 2, and 3 viruses. About the same titres, as determined by haemadsorption in cell cultures, were obtained in LLC-MK2, GMK AH-1, and BSC-1 cells when trypsin had been added to the medium, whereas the Vero cells were less sensitive to the influenza virus strains tested. Virus titres were usually low in the absence of trypsin. A laboratory strain of parainfluenza 2 virus reached about the same titres in medium without as in medium with trypsin, possibly owing to prior adaptation by passages in Vero cells. Comparative titrations of influenza A, and parainfluenza 1 and 3 viruses suggested the same susceptibility of LLC-MK2 cells with trypsin as of primary monkey kidney cells. Re-isolation experiments from 38 clinical specimens showed LLC-MK2 cells to be as efficient as primary monkey kidney cells for isolation of influenza and parainfluenza viruses, whereas the susceptibility of the other cell lines to clinical material has not yet been tested on a larger scale. It is concluded that a continuous line of monkey kidney cell culture may be acceptable as an alternative to primary monkey kidney cells for the isolation of influenza and parainfluenza viruses from patients. PMID- 6274141 TI - Tumour induction in Syrian hamsters fed a combination of aminopyrine and nitrite. PMID- 6274143 TI - Comparative in vitro activity of first, second and third generation cephalosporins. AB - Minimum inhibitory concentrations (MIC) were determined against 662 recent clinical isolates for eight cephalosporins representing first, second and third generation compounds. All four third-generation cephalosporins tested (cefoperaxone, cefotaxime, ceftazidime and moxalactam) were significantly more active against aerobic gram-negative bacteria than the older compounds (cephalothin, cefamandole, cefoxitin, and cefuroxime). Cefotaxime and moxalactam were most active against Enterobacteriaceae with extremely low MIC-values. Ceftazidime was definitely most active against Pseudomonas aeruginosa with more than 90% of strains inhibited at 4 micro g/ml. MIC-values for cefotaxime against Staphylococcus aureus were for all strains 1-2 micro g/ml, slightly higher for cefoperazone, while the effect of ceftazidime and moxalactam was more limited. All third generation cephalosporins demonstrated efficiency against Streptococcus pyogenes, cefotaxime being most active and moxalactam least active, but were essentially ineffective against Streptococcus faecalis. Moxalactam demonstrated higher activity against Bacteroides fragilis than other second and third generation cephalosporins including cefoxitin. Previous studies have demonstrated a very high activity of all third generation cephalosporins against Haemophilus influenzae and Neisseria gonorrhoeae, including beta-lactamase producing strains. PMID- 6274144 TI - Identification and characterization of Kingella kingae. AB - Kingella kingae is a rarely isolated opportunistic pathogen in the family Neisseriaceae. Thirteen strains of this organism, including six strains isolated in Denmark, were characterized. They formed a homogeneous group of small, non motile, fastidious Gram-negative rods which were beta-haemolytic, oxidase positive, catalase negative and saccharolytic. Acid was produced for glucose and maltose. Growth was most pronounced aerobically but corroding colonies increased considerably in size during anaerobic incubation. PMID- 6274145 TI - Enzymatic hydrolysis of immobilized sphingomyelin by three bacterial phospholipases C. AB - Through hydrophobic interaction, sphingomyelin was adsorbed to agarose beads containing octyl groups by a stepwise dilution procedure. This immobilized lipid was used as a substrate for three bacterial phospholipases C (E.C. 3.1.4.3.). The degradation with time of this substrate showed two different fractions of the substrate according to hydrolysing velocity in the early part of the time-curve when phospholipases C from Bacillus cereus and Clostridium perfringens were used. The early fractions could be predigested by the enzymes, a procedure which resulted in linear time-curves. The corresponding early part of the time-curve for phospholipase C from Staphylococcus aureus was linear, indicating a comparatively large early fraction of the substrate for this enzyme. The stock gel of the immobilized lipid substrate could be stored for months. It was easily and reproducibly handled as a water suspension. After enzymatic hydrolysis the substrate was rapidly separated from enzyme and product by filtration. The enzyme assay presented thus represents a convenient way to avoid the difficulties connected with the use of temporary sonicated suspensions as substrate for bacterial phospholipases C. PMID- 6274146 TI - Enzymatic hydrolysis by bacterial phospolipases C and D of immobilized radioactive sphingomyelin and phosphatidylcholine. AB - An assay system for phospholipases C has been described with sphingomyelin immobilized to octyl-Sepharose CL-4B as substrate. The immobilization procedure was further developed and used with [14 C-choline]-sphingomyelin and [14C choline] phosphatidylcholine (lecithin). These immobilized radioactive phospholipids made the enzymatic assays easier to perform and made it possible to increase the sensitivity. Furthermore, since release of the choline part instead of the phosphate part of the substrate molecule was measured, it was possible to use this assay for phospholipase D as well. The enzyme characteristics of phospholipase D from Corynebacterium ovis were compared in this test system with those of three phospholipases C (from Clostridium perfringens, Bacillus cereus and Staphylococcus aureus) with respect to hydrolysing capacities and optimal ion concentrations. PMID- 6274147 TI - Staphylococcal protein-A agglutination assay for avian viruses. AB - Protein-A rich staphylococci were used to develop an agglutination assay for the detection and quantitation of avian reticuloendotheliosis virus (REV) and Herpes virus of turkeys (HVT). Rabbit anti-REV or anti-HVT were used to coat the staphylococci and these reagents were mixed with REV or various other viral antigen preparations. An agglutination titer was determined for antigen preparations of varying purity and the results indicated that in addition to being rapid, this assay allows the detection of as little as 0.3 to 0.5 microgram of partially purified viral antigens. No crossreactivity was observed with the two antisera and the various antigen preparations. This study also confirmed the reports in the literature that chicken immunoglobulins do not bind to protein-A of the Cowan I strain of Staph. aureus. PMID- 6274148 TI - Demonstration of bovine viral diarrhea virus antigen in cryostat- and paraffin sections of bovine tissues by the immunoperoxidase technique. AB - An immunoperoxidase technique (IPT) for the demonstration of bovine viral diarrhea virus (BVDV) antigen in tissue sections is presented. The method was applied to cryostat sections and to alcohol fixed, paraffin embedded specimens. The specificity as well as the sensitivity of the IPT corresponded to the immunofluorescence technique (IFT), and the specimen processing methods used represented an improvement of the histomorphological preservation of tissue structures. PMID- 6274149 TI - Binding of purified C1 subcomponents, C1 inactivator and their complexes to immobilized heparin. AB - Under specified conditions purified C1q, activated C1r and C1s and C1r-C1s complexes were bound independently of Ca2+, to heparin-Sepharose, and could be eluted by an increasing salt gradient. Zymogen C1r and C1s, C1r-C1s complexes, C1 inactivator, and C1r-C1s-C1 activator complexes were not bound. However, at lower conductance Ca2+ independent binding of C14 occurred, which was utilized in the purification of C14 and C1s. In the presence of C1t (serum amyloid P component), C1s was firmly retained on heparin-Sepharose, which was probably due to formation of a C1s-C1t complex. PMID- 6274150 TI - Pulmonary and systemic circulatory responses elicited by hyperosmotic solutions injected into the bronchial artery. AB - In open chest anaesthetized dogs the haemodynamic effects of solutions of equal hyperosmolarity (viz. NaHCO3 8%, NaCl k.6%, and glucose 34.3%, solutions) given into the bronchial artery were studied. Administration of any of these solutions directly into the bronchial artery resulted in increased cardiac output, stroke volume, bronchial blood flow, and bronchial fraction of the cardiac output, and decreased heart rate and bronchial as well as pulmonary vascular resistances. When given into the pulmonary circulation, the same solutions evoked similar reactions of smaller magnitude. To exclude the effect of major surgical trauma and the open-chest condition, another experimental model closer to the physiological situation was also developed. In this preparation NaHCO3 failed to produce the above haemodynamic response even when given into the bronchial artery. After a one-hour bleeding period resulting in a drop of arterial blood pressure to 40 mmHg, while using the same preparation, the administration f NaHCO3 solution into the bronchial artery caused a significant rise in blood pressure in both the systemic and pulmonary arteries. In these experiments a correlation was found between arterial oxygen tension and the extent of change in blood pressure. The exact mechanism of action of the observed haemodynamic changes is still not clear. However, it is likely that receptors localized in the area of the bronchial circulation and sensitive to hypoxia might have played a role in the development of the haemodynamic effects described. PMID- 6274151 TI - Effects of drugs influencing the cAMP level on hippocampal seizure activity. AB - Changes of hippocampal EEF seizure activity elicited by electrical stimulation of the hippocampus or penicillin injection into the hippocampus were investigated under the effect of locally applied drugs influencing the cAMP level in the brain. It was observed that some drugs which elevate the cAMP level such as papaverine, histamine +K+ and dibutyryl cAMP elevated the electric seizure threshold while in penicillin-induced epilepsy they reduced the occurrence of ictal activity and the interictal spike frequency. These drugs when applied before penicillin prolonged the time necessary for development of the epileptic focus. The effect of imidazole was the opposite in every respect. On the basis of these data the possible role of cAMP in the pathomechanism of epilepsy is discussed. PMID- 6274152 TI - Comparative quantitative estimation of the granular vesicles in the intramural ganglia of Auerbach's plexus in the nonsphincter and sphincter parts of cat alimentary tract. AB - The spatial size and density of the granular vesicles (GV) in the axonal varicosities of the intramural ganglia in parts of the alimentary tract adjacent to the lower esophageal, pyloric and ileo-caecal sphincters are determined. The results obtained are compared with the data about the density and size of GV in these sphincters, described earlier (Lolova et al., 1980). The results concerning GV size show: (1) absence of very small differences in GV diameters from the 1st and 2nd populations (average diameter around 40, respectively 55 nm) between the sphincter and nonsphincter parts; (2) absence of 3rd population GV (average diameter 77 and 90 nm) in the nonsphincter parts of the fundus close to the terminal ileum; (3) GV from 4th and 5th populations (average diameter around 125 nm, 150 nm respectively) are larger in the nonsphincter parts than in the sphincters. The following observations are made with respect to GV density: (1) the smallest GV have the greatest density in all parts studied; (2) the GV density from the different populations differs in the nonsphincter and sphincter parts; (3) certain characteristic features become prominent upon comparing GV density in the parts of the gastro-intestinal tract from cranial to distal direction. The results obtained are discussed with a view to the functional characteristics of the nonsphincter and sphincter parts and with the participation of various transmitters in them. PMID- 6274153 TI - The cyclic AMP level during an increased blood flow in cat cerebral cortex. AB - The experiments were carried out on cats anesthetized with chloralose, curarized and artificially ventilated. The local cortical CBF was recorded by the thermoclearance technique. The cAMP level in the grey matter of the cerebral cortex and the arterial blood pressure, as well as the arterial blood gas content were measured. Increased concentration of CO2 in the inspired mixture (paCO2 = 79.8 +/- 8.5 mm Hg) caused an enhancement of CBF, without inducing changes in the cAMP level (1.71 +/- 0.07 nmoles/g wet weight). Increasing the PaCO2 up to 184.42 +/- 12.3 mm Hg produced neither a further increase of CBF, nor a rise of cAMP level. Adrenaline injected into the cerebral ventricles led to an increase of cAMP up to 4.42 +/- 0.7 nmoles/g without changing CBF. Saline in the same volume increased cAMP level to a smaller degree (2.71 +/- 0.26 nmoles/g). No changes in CBF were registered. Dibutyryl cAMP, cAMP and adenosine injected into a. lingualis did not significantly change the CBF. PMID- 6274154 TI - Pharmacological and biochemical study in ontogenetic aspect of 3'-5' adenosinemonophosphate phosphodiesterases with low and high Km. AB - The ontogenetic development of 3',5'-AMP phosphatediesterase with low and high Km is studied in rat ileum and brain, and more specifically the changes in the kinetic parameters of the enzyme. Comparison of the Michaelis constants of the phosphodiesterase (PDE) in the ileum and brain of newborn and adult rats reveals a rise in their values in the ileum and a decrease in the brain. It may be assumed that the development of PDE with low Km in the ileum precedes that in the brain, which is directly related to the sequence in the postnatal development of the adaptational mechanisms to the environment in the young organism. In studies on the relation of PDE in the ileum and brain to some inhibitors (papaverine, theophylline, 6'-iodo-dihydropapaverine and 6'-bromo-dihydropapaverine) we established mosaic effects of these substances both with respect to the type of the enzyme and with respect to the age of the experimental animal. The results of our kinetic experiments showed that there was no difference in the type of inhibition and in the correlation of the inhibitory power of the drugs tested during ontogenesis. Probably no essential changes occur in the active centres of the enzymes during this period. The observed age changes in PDE with low and high Km condition the altered sensitivity of these enzymes to the inhibitors studied. The enzyme in the ileum of newborn animals--and organ with earlier functional maturation--is more sensitive to inhibitors. Conversely, PDE sensitivity in the brain increases during the ontogenesis. PMID- 6274155 TI - Load-dependent activation of calcium turnover and cardiac performance in isolated rat atria. AB - The effects of increasing load on developed tension (DT) and its second derivative, 45Ca uptake, as well as on the levels of cAMP, cGMP, PGE and PGF2 alpha were investigated in spontaneously beating isolated rat atria. Gradually increasing loads augmented, parallelly, both DT and 45Ca uptake into the atria. The levels of cAMP, cGMP, PGE and PGF2 alpha were not altered. Indomethacin (1 microM) did not antagonize the effect of loading on DT. It is concluded that activation of calcium mobilization is involved in the length-tension relationship of the heart. This mobilization is not connected with changes in tissue concentrations of cAMP or of prostaglandins. PMID- 6274156 TI - Influence of adenosine on responses to vagal nerve stimulation in the anesthetized rabbit. AB - The influence of local adenosine infusion into the celiac artery on the gastric contractile responses to centrifugal vagal nerve stimulation was studied in anesthetized rabbits, and was compared with the effects of systemic administration of equivalent amounts of adenosine. Close arterial infusion of adenosine caused a marked reduction of gastric contractions induced by nerve stimulation, whereas corresponding responses to close arterial infusions of acetylcholine were enhanced during adenosine. The comparison with systemic adenosine administration revealed that the influence on gastric neurotransmission was not related to the hypotensive effect of the compound. No effects of adenosine were seen on bronchial activity as measured by insufflation pressure. Variable effects were obtained on cardiac responses to vagal stimulation. Gastric smooth muscle contractions elicited in vitro by transmural nerve stimulation were affected by adenosine in a biphasic manner, initial inhibition followed by potentiation of the apparently cholinergic responses. It is suggested that adenosine may modulate cholinergic neurotransmission in vivo by a dual effect, prejunctional inhibition and postjunctional enhancement. PMID- 6274157 TI - Theophylline interferes with the modulatory role of endogenous adenosine on cholinergic neurotransmission in guinea pig ileum. AB - The ability of theophylline and other phosphodiesterase inhibitors to alter contractile responses to cholinergic nerve stimulation was investigated in isolated longitudinal muscle of the guinea pig ileum. Theophylline in low concentrations (10-100 microM), having no or little effect on measured phosphodiesterase activity, antagonized inhibitory effects of exogenous adenosine. In higher concentrations (0.1-10 mM), shown to be effective in inhibiting phosphodiesterase, theophylline as well as a "pure" cAMP phosphodiesterase inhibitor, ZK 62, 711, inhibited contractile responses. Dipyridamole and dilazep, inhibitors of adenosine inactivation, and also selective inhibitors of cAMP and cGMP phosphodiesterase, respectively, were found to enhance effects of exogenous adenosine and to cause a marked leftward shift to adenosine threshold dose. When dipyridamole and dilazep by themselves had inhibitory effects these could be antagonized by theophylline, suggesting an action through increased levels of endogenous adenosine. As a further indication of endogenous adenosine modulating neurotransmission low concentrations of theophylline enhanced responses to transmural stimulation. Endogenous purine concentrations in tissues and bath media were measured by HPLC. Because of tissue and microbial adenosine inactivation direct estimates of extracellular adenosine concentration could not be obtained. However, adenosine levels increased during transmural stimulation, and during inhibition of adenosine inactivation were sufficient, even in the bath medium, to interfere with the cholinergic neurotransmission. PMID- 6274158 TI - The effect of temperature elevation on the cerebrovascular response to noradrenaline and 5-hydroxytryptamine. AB - Fever frequently complicates stroke and subarachnoid haemorrhage. A transient rise in transmitter monoamine levels of plasma and cerebrospinal fluid occurs in these diseases. The present study demonstrates an enhanced vasoconstrictor response of cerebral vessels to noradrenaline-but not 5-hydroxytryptamine following a rise in temperature. The augmented response is more likely due to an impaired inactivation (re-uptake) of the amine than to an altered sensitivity of the post-synaptic alpha-adrenergic receptor, since it could be reproduced by pretreatment with cocaine. The finding indicates that it may be important to combat fever in these patients. PMID- 6274159 TI - Cyclic AMP-dependent and -independent effects of prostaglandins on the contraction-relaxation cycle of spontaneously beating isolated rat atria. AB - The effects of prostaglandin (PG) F2 alpha, E2, E1 and I2 on the amplitude, duration of the contraction-relaxation cycle (CRC), the second derivative of developed tension and the cyclic adenosine-3', 5' monophosphate (cAMP) level and on 45Ca uptake were studied in isolated spontaneously beating rat atria. The order of capacity to generate positive inotropic effects was PGF2 alpha greater than PGE2 greater than PGE1 approximately PGI2. Only PGI2 and PGE1 decreased the duration of CRC. PGF2 alpha produced an increase during the first 2.5 min, whereafter the duration returned to the initial level, PGE2 had no significant effect on the shape of the CRC. The ratios of the maximum and minimum of the second derivative of the developed tension were reduced by PGI2 and PGF2 alpha 2.5 and 5 min after administration, respectively. The 45Ca uptake was stimulated equally by all of the tested PGs, but only PGI2 and PGE1 could significantly increase the cAMP level. The results do not support the conception that cAMP could mediate the positive inotropic effect of PGs. Rather the contrary, cAMP, increased by PGE1 or PGI2, could be responsible for increased relaxation, which might prevent the full development of tension. PMID- 6274160 TI - Immunoreactive ACTH in rat pineal: stress and dexamethasone lower the concentrations. PMID- 6274161 TI - Preponderance for either alpha- or beta-adrenoceptor mediated sensitization in the rat submaxillary gland. AB - The sensitivity of the rat submaxillary gland was examined 3-4 weeks after either parasympathetic decentralization or sympathetic decentralization or denervation. The threshold doses for secretion of saliva of parasympathomimetic (methacholine) and sympathomimetic (noradrenaline, adrenaline, phenylephrine and isoprenaline) drugs were estimated and the amount of saliva secreted in response to supraliminal doses of these drugs was measured. Each type of operation caused the development of a supersensitivity that involved all three types of receptors, i.e. muscarinic cholinoceptors, alpha-adrenoceptors and beta-adrenoceptors. Following parasympathetic decentralization the sensitization was predominantly mediated via alpha-adrenoceptors, and also via cholinoceptors. Following sympathetic decentralization or denervation the postjunctional sensitization was predominantly mediated via beta-adrenoceptors; most of the supersensitivity to noradrenaline, adrenaline and phenylephrine found after sympathetic denervation was of the prejunctional type. An increase in receptor density and an intracellular arrangement where the response of cholinoceptors and alpha adrenoceptors is mediated via one pathway and the response of beta-adrenoceptors via another are suggested as factors that may be of importance for the development of the postjunctional supersensitivity. The present study shows that the traffic of secretory impulses in the sympathetic nerve is of importance for the level of sensitivity of the secretory cells. Since postjunctional supersensitivity following sympathetic denervation did not exceed that following sympathetic decentralization it is suggested that under normal conditions a continuous release of noradrenaline from the nerve endings is of little importance for the level of sensitivity. PMID- 6274162 TI - Is chloride transfer in frog skin localized to a special cell type? PMID- 6274163 TI - Early changes in ATP and cyclic AMP levels in experimental critical skin flaps. AB - A cranially based pedicle skin flap, 2 cm wide and 7 cm long, was prepared in the midline of the rat dorsum. The contents of ATP and cyclic AMP were determined in the proximal, middle and distal parts of the flap. The basal level of ATP was 1.3 +/- 0.1 nmol/g dry weight. Following the operation the level of ATP fell rapidly. The fall was more marked in those parts of the flap that eventually do not survive. During the first 12 h the level of cyclic AMP fell at least as rapidly. However, thereafter the cyclic AMP level remained constant or even increased while the ATP showed a further decrease. It is suggested that the rapid fall in ATP may be used to assess the eventual survival of skin flap. The results are also compatible with the opinion that a period of metabolic stimulation in the skin flap starts 12--18 h after operation, i.e. at the time when the adrenergic nerves in the flap degenerate. PMID- 6274164 TI - Classification of beta-adrenoceptors in the microcirculation of skeletal muscle. AB - Nervous and humoral beta-adrenergic, postjunctional effects on microvascular resistance, on precapillary sphincter tone, and on transcapillary fluid exchange in cat skeletal muscle (Lundvall & Jarhult 1974, 1976 a, Lundvall & Hillman 1978 a, b) were evaluated with regard to the beta 1- or beta 2-specificity of the adrenoceptors. Marked beta 2-dilator responses but no significant beta 1-effects were observed. The conclusion was therefore reached that neurogenic as well as humoral beta-adrenergic control of the microcirculation in skeletal muscle is exerted via activation of beta 2-adrenoceptors. PMID- 6274165 TI - Effects of ACTH and alprenolol treatments on muscle and brown fat enzyme activities and weights in the rat. AB - Chronic ACTH treatment (3 IU/kg daily for 8 days) caused an increase in wet weight of the interscapular brown adipose tissue (ISBAT) in the rat without affecting the enzyme activities of the cardiac or skeletal muscles or of ISBAT. On the other hand, chronic beta-blockade (alprenolol 10 mg/kg daily for 8 days) induced elevated activities in the oxidative enzymes of the ISBAT but not of the muscle tissues measured. Combined ACTH and alprenolol treatment also increased significantly the citrate synthase and malate dehydrogenase activities and protein concentration, but decreased the weight of ISBAT to normal. The results suggest that, although a direct antagonism between the beta-blockers and ACTH has not been demonstrated, beta-blockers can abolish the ACTH-induced weight gain of the ISBAT. PMID- 6274166 TI - Acute effects of gonadotrophins and cyclic AMP on protein synthesis and progesterone production by isolated rat granulosa cells. AB - Amino acid uptake, protein synthesis and progesterone production were studied in rat granulosa cells, isolated from follicles of different stages of development. The amino acid uptake in granulosa cells from prepubertal rats was rapid with a distribution ratio above 1 within 10 min. No significant effects of gonadotrophins were observed on this parameter. The acute influence of exogenous LH and FSH or dibutyryl cyclic AMP (dbcAMP) on incorporation of 3H-leucine or 3H phenylalanine was investigated as a measure of the rate of protein synthesis. In most experiments progesterone production was determined concomitantly. Both FSH (10 micrograms/ml or 100 micrograms/100 g b.wt.) and dbcAMP (a mM) stimulated (1.6--1.8 fold) the incorporation of leucine into granulosa cell proteins from prepubertal rats while LH was without effect. Progesterone production in these granulosa cells was very low and neither the gonadotrophins nor dbcAMP were stimulatory. Similarly designed experiments were performed on granulosa cells isolated from preovulatory rat follicles. It was then found that both FSH (10 micrograms/ml or 100 micrograms/199 g bst.) and dbcAMP (1 and 5 mM) as well as LH (10 micrograms/ml or 100 micrograms/100 g b.wt.) significantly (1.2--2 fold) stimulated protein synthesis. Furthermore, basal progesterone production was higher and was markedly stimulated (3--6 fold) by all three substances tested. The observations that the protein synthesis in immature granulosa cells is increased by exposure to FSH and dbcAMP while progesterone production in not, whereas both these parameters are stimulated with FSH, LH and dbcAMP in the preovulatory granulosa cells suggest that there might be certain differences in the nature of the proteins synthesized by immature and preovulatory granulosa cells. PMID- 6274167 TI - Long-term preservation of transfecting activity of avian sarcoma proviral DNA. AB - The integrated proviral DNA of avian sarcoma virus (ASV) in host cell chromosomes has been isolated and stored in saline sodium citrate (SSC) solution or in 70% ethanol at 4 degrees C in a refrigerator over 4 years. This DNA was assayed by transfection of chick embryo cells(CEC). The biological activity of cellular transformation by the stored DNA was compared with that of a fresh isolate of the proviral DNA. The efficiency of the transfection by each DNA was almost the same. PMID- 6274168 TI - Microinjection of nucleic acids into cultured mammalian cells by electrophoresis. AB - Simian virus 40 (SV40) DNA was microinjected into cultured mammalian cells by means of electrophoresis (iontophoresis). Successful transfer of DNA into cells was confirmed by detecting SV40 T antigen using the indirect immunofluorescent technique. PMID- 6274169 TI - Relationship between circulating immune complexes and angiotensin-converting enzyme in pulmonary sarcoidosis. AB - The relationship between circulating immune complexes (CIC), serum angiotensin converting enzyme (SACE) and clinical features was investigated in 119 patients with sarcoidosis. CIC, measured by the C1q-binding test and a polyethylene glycol precipitation test, were detected in 42% of the patients. A significantly higher level of CIC was found in patients with duration of the disease of more than 2 years and in patients in stage II or III on chest X-ray. SACE was increased in 45% of the patients, most frequently in those with active disease and in those in stage II or III on chest X-ray. No positive correlation was found between CIC and elevated levels of SACE. PMID- 6274170 TI - [Case of Madurella grisea maduromycosis. Study of the strain]. PMID- 6274171 TI - Brain serotonin receptors and neuropsychiatric disorders. PMID- 6274172 TI - Role of serotonin in the hypothalamic regulation of pituitary function. AB - Serotonin is only one of numerous neurotransmitters that may have roles in the regulation of pituitary secretion. The literature pertaining to serotoninergic involvement in the hypothalamic control is large, and a report of this type cannot provide a comprehensive review of that literature. Instead, the data presented and the literature cited were intended to be illustrative of the use of drug intervention and other techniques for probing the role of serotonin in the regulation of pituitary secretion. Clearly further studies with each of the pituitary hormones are needed, and the development of drugs acting on serotonin neurons with greater specificity can be a strong impetus to progress in this research area. PMID- 6274173 TI - [Clinical and etiological study of adenovirus conjunctivitis, 1979-1980, Sapporo, Japan: with special reference to the infection with types 4 and 19 (author's transl)]. PMID- 6274174 TI - [Effect of detergent-soluble extract of virus-infected cells on preestablished herpetic latency in the trigeminal ganglia on mice. Preliminary report (author's transl)]. PMID- 6274175 TI - Critical values for group system acid phosphatase (AP), phosphoglucomutase and (PGM) adenylate kinase (AK). PMID- 6274176 TI - [Wilm's tumor in adults and Grawitz' tumor in children]. PMID- 6274177 TI - [2 cases of renal metastasis. Rare etiology, diagnostic pitfalls and radiographic symptoms]. PMID- 6274178 TI - Cytologic bile examination in the diagnosis of biliary duct neoplastic strictures. AB - Cytodiagnosis of the bile is not frequently thought of as a method of obtaining a histopathologic diagnosis of the cause of a neoplastic biliary duct stricture. However, cells surrounding the biliary ducts are continuously exfoliated into bile and become available for cytologic examination whenever bile is collected. Nineteen patients with obstructive jaundice are reported. In seven of 15 with neoplastic biliary duct strictures, cytodiagnostic examination showed tumor cells. There were no false-positive results. The sensitivity of bile cytodiagnosis in this series was 47%, its specificity was 100%, and its accuracy was 58%. PMID- 6274179 TI - CT diagnosis of tumor thrombus in the inferior vena cava: avoiding the false positive diagnosis. PMID- 6274180 TI - Contrast enhancement of the irradiated spinal cord in children. AB - Four children are reported in whom marked contrast enhancement of the spinal cord and roots was demonstrated by computed tomography months to years after relatively low dose therapeutic irradiation of paraspinal tumors, the radiation field including the cord. This phenomenon, previously unreported, probably represents subclinical radiation injury. None of the children had any neurologic abnormalities. PMID- 6274181 TI - Air sampling in the assessment of continuous exposures to acutely-toxic chemicals. Part I--Strategy. AB - This paper describes the conceptual basis for assessing the hazard of continuous exposures to acutely toxic chemicals. It is stressed that the variability of workers' short-term exposures to airborne chemicals must be considered in establishing strategic criteria which define an acceptable (or unacceptable) environment. The argument is developed that the traditional strategy of an absolute ceiling limit is unworkable since any set of exposures can be declared out of compliance if a sufficiently large number of samples is collected. It is proposed that acute-exposure limits be defined instead as air concentrations that can be exceeded by an explicit fraction of the population of exposures. An "exceedance" rate of 5% is suggested provided that exposure limits incorporate appropriate safety factors which are relatively large for potent acute toxins and small for irritants. The implications of this strategy are analyzed with some existing ceiling limits proposed by NIOSH. PMID- 6274182 TI - Improvement in supine bicycle exercise performance in refractory congestive heart failure after isosorbide dinitrate: radionuclide and hemodynamic evaluation of acute effects. PMID- 6274183 TI - Hemodynamic correlates of right ventricular ejection fraction measured with gated radionuclide angiography. AB - Right ventricular function was studied in 60 patients with equilibrium gated radionuclide angiography. The mean (+/- standard deviation) right ventricular ejection fraction in 20 normal subjects was 53 +/- 6 percent, a value in agreement with previous data from both radionuclide and contrast angiographic studies. This value was similar (55 +/- 7 percent) in 11 patients with coronary artery disease but normal left ventricular function. Radionuclide measurements of right ventricular ejection fraction were correlated with right heart hemodynamics. There was a significant negative linear correlation between right ventricular ejection fraction and mean pulmonary arterial pressure (r = -0.82) and between right ventricular ejection fraction and right ventricular end diastolic pressure (4 = -0.67). Furthermore, patients with elevated right ventricular end-diastolic pressure and mean pulmonary arterial pressure had a more severely depressed ejection fraction than did those with an elevated mean pulmonary arterial pressure alone. Thus, an abnormal value for right ventricular ejection fraction by gated radionuclide angiography in the absence of primary right ventricular volume overload suggests abnormal right heart pressures, whereas a normal value excludes severe pulmonary arterial hypertension or an elevated right ventricular end-diastolic pressure. PMID- 6274184 TI - Nongeometric determination of right ventricular volumes from equilibrium blood pool scans. PMID- 6274185 TI - Colon ulceration in lethal cytomegalovirus infection. AB - Six renal transplant recipients with severe cytomegalovirus (CMV) infection developed colonic ulceration and lower gastrointestinal bleeding. All patients died between four and 84 days following onset of bleeding. Four patients required colon resection to control blood loss. In the remaining two cases, there was complete autopsy examination with thorough tissue sampling of the colon. In addition to routine light microscopic examination, all cases were studied with an immunoperoxidase technic for identification of CMV. Electron microscopic examination of the colon was performed on two cases. In this select group of patents, CMV plays an active role in damaging colonic mucosa, primarily as a result of CMC vasculitis. Clinical and pathologic features of our cases are compared to published reports of CMV infection of the colon. PMID- 6274186 TI - A thoracic benign mesenchymoma in association with hemihypertrophy. AB - A thoracic benign mesenchymoma was removed from an 18-year old caucasian male who also had hemihypertrophy. The very large benign mesenchymoma which was removed was found to consist of an admixture of cytologically mature vascular spaces, adipose tissue, and smooth muscle. After removal of the tumor, the patient has had no recurrence with one year of follow-up. Patients with hemihypertrophy appear to be at high risk to develop neoplasms. Malignant neoplasms of the adrenal gland, kidney, and liver are most common. It is suggested that patients with hemihypertrophy should be followed periodically for their entire life because of their propensity to develop tumors. PMID- 6274187 TI - Falsely normal value in fluorometric transferase screening of galactosemic blood. A cautionary note. AB - A blood sample from a galactosemic infant gave a normal result with the fluorescent screening test for galactose-1-phosphate uridyl transferase. The generation of fluorescence from this sample was found to be property of the plasma; the erythrocytes manifested no galactose-1-phosphate uridyl transferase activity. The plasma was found to contain a high level of isocitrate dehydrogenase, presumably a result of the infant's liver disease, and of isocitrate, presumably derived from citrate in the anticoagulant solution by action of aconitase derived from leukocytes which lysed during shipment. The oxidation of the isocitrate by the dehydrogenase apparently resulted in reduction of NADP to NADPH. False negative results have not been reported previously using the fluorescent transferase assay. They can be avoided in the future by using EDTA rather than citrate as an anticoagulant, particularly if blood samples are to be shipped at ambient temperature. PMID- 6274188 TI - Gastrointestinal structure and function in Fabry's disease. AB - We evaluated gastrointestinal structure and function in 13 hemizygous males and 17 heterozygous females, five to 67 years old, from four kindreds with Fabry's disease. Gastrointestinal symptoms, noted in 62% (8/13) of the males and 29% (5/17) of the females, were present prior to the diagnosis of Fabry's disease in five patients; were discovered at the time of study in six patients and were associated with multiple other symptoms in two patients. Serum protein, albumin, folate, Vitamin B12, calcium, phosphorous, cholesterol and iron were normal in all 30 patients. Xylose absorption was normal in 2/2 males and 13/13 females studied. HLA B8 antigen was present in none of the males and 2/17 females. Peroral duodenal (one male), jejunal (six males, two females) and rectal (one male) biopsies on light microscopy demonstrated a normal villous pattern and luxol-fast blue positive "foamy" cell deposits in all males, while no deposits were visualized in the females. In all males and females studied, electron microscopic examination showed electron dense, intralysosomal "zebra-like" (0.5 0.75 micrometer.) bodies in the vascular endothelial and perithelial cells and in the cytoplasm of the small unmyelinated neurons, and perineurial cells. Despite the frequency of gastrointestinal symptoms, both malabsorption and celiac disease were absent. PMID- 6274189 TI - Evidence for the generation of hydroxyl radical during arachidonic acid metabolism by human platelets. AB - Reactive oxygen species, probably hydroxyl radicals (OH.), have been suggested to be generated during arachidonic acid (AA) metabolism and, once released, these species can modify the rate and extent of various reactions involved in AA metabolism. We have studied this phenomenon in washed human platelets. OH. generation was quantitated using 14C-benzoic acid as a specific trap in a continuous ionization chamber system. Resting platelets did not produce any detectable signal, whereas addition of AA resulted in gradual OH. production with peak values detected at approximately 20 min. Similar studies conducted under nitrogen or after boiling the platelets almost abolished OH. generation. Aspirin had no significant effect, whereas 5,8,11,14-eicosatetraynoic acid decreased the signal by greater than 90%, thus suggesting that OH. is produced primarily through the lipoxygenase pathway. Superoxide dismutase (SOD) and catalase had no effect and, as expected, phenol and mannitol decreased OH. production considerably, by greater than 50% and 90%, respectively. Azide and cyanide also reduced the OH. generation by about two thirds. We conclude that OH. is generated during AA metabolism by human platelets. It is primarily produced via the lipoxygenase pathway and may require a heme-dependent peroxidase. This highly reactive oxidant may play an important role in normal and abnormal hemostasis. PMID- 6274190 TI - Fatal disseminated adenovirus 11 pneumonia in an agammaglobulinemic patient. AB - Adenovirus type 11, an organism not previously associated with pneumonia, caused the death of a patient with infantile x-linked agammaglobulinemia who had normal cell-mediated immunity. Despite long-standing, regular therapy with immune globulin, his serum lacked neutralizing antibody to the virus. This case confronts the conventional view that viral infections are primarily resisted by cellular immune reactions and reemphasizes the importance of antibody in the host defense against adenoviruses. It further demonstrates the continued vulnerability of such patients to certain pathogens in the presence of presumably adequate standard-dose passive immunization. PMID- 6274191 TI - Heterozygote detection in Fabry disease utilizing multiple enzyme activities. AB - In Fabry disease, as in other X-linked traits, identification of all heterozygotes is difficult. Reduced plasma alpha-galactosidase activities will correctly identify 60-70% of the carriers. The identification rate improves when an alpha/beta-galactosidase activity enzyme ratio is used. We measured alpha galactosidase activity in reference to several other enzyme activities, beta galactosidase, beta-hexosaminidase, and alpha-fucosidase in plasma and leukocytes from 22 suspected and 9 obligate carriers from 4 kindreds of Fabry disease patients. Utilizing such ratios or various combinations of ratios in plasma we have correctly identified the carrier state in 91% of heterozygotes. Leukocyte alpha/beta-galactosidase identified one more female than leukocyte alpha galactosidase activities alone. We recommend the use of such multiple biochemical tests to identify carriers of Fabry disease. PMID- 6274192 TI - Responses of the pelvic vascular bed to intra-arterial stimulation of beta adrenergic and cholinergic receptors in pregnant and nonpregnant sheep. AB - The responses of different parts of the pelvic vascular bed to beta-adrenergic and cholinergic receptor stimulation were studied in chronically instrumented, unanesthetized pregnant and nonpregnant sheep. Isoproterenol and acetylcholine were administered intra-arterially in progressively increasing bolus injections, and dose-response curves were constructed for changes in blood flows, arterial pressure, and heart rate. Response to beta-adrenergic and cholinergic stimulation was measured before and after the agents reached the central circulation. In addition, data were obtained from the same animals before and after pharmacologic cardiac denervation. Results show: (1) Stimulation of beta-adrenergic and cholinergic receptors produced active vasodilatation in extrauterine vascular beds with minor changes in uterine blood flow of the pregnant horn; (2) the changes in uterine flow observed during beta-adrenergic and cholinergic stimulation were probably secondary and related to a shift of blood from the uterus to other vascular beds that were actively dilated; (3) the response of the pregnant animal to any given dose of the autonomic agonist was strikingly smaller to that of the nonpregnant animal. A number of physiologic factors peculiar to pregnancy may contribute to this difference; among these are: (1) dilution factor related to the greater volume flow; (2) near-maximal dilatation of some blood vessels and the presence of the two resistance system of placental circulation, and (3) the effect of progesterone on the vascular smooth muscle. PMID- 6274193 TI - Oxytocin and the initiation of human parturition. I. Prostaglandin release during induction of labor by oxytocin. AB - Concentrations of plasma prostaglandins E and F and the 15-keto-13,14 dihydrometabolite of PGF2 alpha (PGFM) were determined by radioimmunoassay in 15 women who underwent induction of labor with oxytocin. Plasma PGFM rose significantly during the oxytocin infusion in nine women who went on to deliver vaginally but did not change in six women in whom induction of labor failed. Plasma PGE and PGF levels also rose during the infusion in the nine women with successful induction of labor but the changes were not statistically significant. In comparison to the six women in whom induction failed, however, plasma PGE in the nine women with successful induction reached significantly higher levels. Oxytocin infusions elicited uterine contractions of similar frequency in both groups of women, but the cervix failed to dilate in the six women in whom induction failed. The oxytocin-induced rise in plasma PGFM is, therefore, not simply a consequence of uterine contractions. We suggest that oxytocin stimulates PGF production in the pregnant uterus when it is appropriately sensitized to oxytocin, causing a potentiation of the oxytocin-induced contractions which is necessary for the contractions to become efficient in dilating the cervix. We further suggest that the stimulation of PGF production by oxytocin is mediated by oxytocin receptors, probably in the decidua. PMID- 6274194 TI - Management of genital herpes simplex virus infection occurring during pregnancy. AB - Genital herpes simplex virus (HSV) infection during pregnancy has caused considerable concern among lay and professional personnel in the past 10 years. Knowledge concerning the potential hazards of HSV to the newborn infant has increased the use of cesarean section for women who have or are suspected of having genital HSV infection near or at the time of labor. Because of this, a 57 month prospective study was begun at Vanderbilt University Hospital and its affiliate, Nashville General Hospital, January 1, 1976, and involved HSV culturing of all suspected genital herpes lesions during pregnancy. Those patients with positive HSV cultures prior to labor and without a subsequent negative culture underwent cesarean section. Those patients whose cultures reverted to negative were considered candidates for vaginal delivery. During the study period, there were 16,381 deliveries at the two institutions. One hundred twenty pregnant women were cultured, with 80 being HSV culture negative and 40 culture positive. The outcome of these pregnancies as well as a review of the experience with neonatal HSV infection in Middle Tennessee is presented. It is concluded that by utilizing HSV cultures of genital lesions as a guide to determining the route of delivery, the incidence of cesarean sections and neonatal HSV infection can be kept to a minimum. PMID- 6274195 TI - Simulation of the follicular phase of the menstrual cycle by intravenous administration of low-dose pulsatile gonadotropin-releasing hormone. PMID- 6274196 TI - Autonomic control of the pelvic circulation: in vivo and in vitro studies in pregnant and nonpregnant sheep. AB - Pregnancy alters the autonomic control of the peripheral circulation. Present in vivo and in vitro studies were designed to investigate the mechanisms of these alterations. In the in vivo studies, pelvic and systemic circulatory responses to intra-arterial and intravenous injections of agonists were monitored in unanesthetized pregnant and nonpregnant ewes. In in vitro studies, a comparison was made of responses to transmural nerve stimulation (TNS) and to norepinephrine (NE) of different blood vessels obtained from pregnant and nonpregnant ewes. The results were the following: (1) Pelvic vascular response to all vasoactive stimuli was considerably more depressed during pregnancy than was systemic circulatory response. (2) In vitro reactivity to alpha-adrenergic stimulation with NE of different blood vessels obtained from pregnant animals was similar to that of vessels from nonpregnant animals. (3) The response to TNS of blood vessels obtained from pregnant ewes was less than that of vessels from nonpregnant ewes. The conclusions were that (a) probably largely related to dilution of a given dose of the stimulus in the greater blood volume and flow; and (b) the increased neurogenic tone of the peripheral circulation during pregnancy is most likely related to increased nerve firing rather than neural density in the vessel walls. PMID- 6274197 TI - Structural relationship between desmosomes and mitochondria in human livers exhibiting a wide range of diseases. AB - Liver biopsies from ten patients (five women and five men, aged 25-63) with a number of different diseases were studied with a transmission electron microscope. In addition to many different pathologic changes in the hepatocytes, all clinical diagnoses showed 5-70% of mitochondria with paracrystalline inclusions. A peculiar finding was that the desmosomes that join two cells had mitochondria associated with the intracytoplasmic component in both cells in 5 15% of desmosomes observed. In addition, the association involved only one side in one cell in 7-23% of desmosomes observed. Only speculation can be made regarding the function of these mitochondrial-desmosomal associations. PMID- 6274198 TI - Glycogen accumulation in alveolar type II cells in 3-methylindole--induced pulmonary edema in goats. AB - The present study shows that intravenous infusion of 3-methylindole (3MI) induced acute pulmonary edema in goats. Edematous changes were seen in the alveoli and the interalveolar interstitium. At 72 hours after treatment, an accumulation of glycogen that had a pathognomonic appearance of alpha particles was observed in the alveolar Type II cells. A rich accumulation of glycogen particles and defective lamellar bodies containing triglycerides were the significant morphologic changes in the alveolar Type II cells. These findings suggest that massive glycogen deposition in the alveolar Type II cells is a defect that might interfere with surfactant production, further complicating the disease process. PMID- 6274200 TI - Giant dense bodies in fibroblasts cultured from beige mice with the Chediak Higashi syndrome. AB - Fibroblasts cultured from the skin of beige mice manifesting the Chediak-Higashi syndrome (CHS), unlike cells derived from normal black mice, exhibited giant dense bodies in the cytoplasm. These megabodies were membrane-delimited and exhibited dense content by electron microscopy, with myelin figures, highly osmiophilic, thick membranous contours, and lucent areas. The megabodies evidenced acid phosphatase ultrastructurally. Cells of both normal and CHS mice contained smaller dense bodies. During a 2--6 hour exposure to colloidal gold, the smaller dense bodies of normal and CHS fibroblasts selectively incorporated the gold spherules and, accordingly, were identified as secondary lysosomes of heterophagic origin. With longer exposure to colloidal gold, the small dense bodies of the normal and CHS cells disclosed increased content of colloidal gold. After 24 hour exposure to colloidal gold, many giant dense bodies also exhibited gold particles, evidencing uptake of endocytosed material into the giant structures and the heterophagic origin of at least some of the content of the bodies. The gold spherules initially incorporated into the giant dense bodies were concentrated in foci along their periphery and indicated fusion of small dense bodies into the giant structures. Transformed normal and CHS cells appeared to contain more abundant myelin figures than nontransformed cells, and these were larger in transformed CHS cells and constituted a major component of their giant dense bodies. The giant inclusions of the transformed CHS cells generally contained little colloidal gold, suggesting their derivation principally through cellular autophagy. PMID- 6274199 TI - Neutrophil-degranulating action of 5,12-dihydroxy-6,8,10,14-eicosatetraenoic acid and 1-O-alkyl-2-O-acetyl-sn-glycero-3-phosphocholine. Comparison with other degranulating agents. AB - 5,12-Dihydroxy-6,8,10,14-eicosatetraenoic acid was prepared from rabbit neutrophils challenged with ionophore A23187 plus arachidonic acid. It was purified by reverse-phase high-performance liquid chromatography with the use of a two-step procedure that effectively resolved the lipid from closely eluting contaminants. The phospholipid 1-O-alkyl-2-O-acetyl-sn-glycero-3-phosphocholine was prepared from beef heart plasmalogen. Confirming earlier reports, both lipids stimulated human polymorphonuclear neutrophils to release granule-bound enzymes. Their respective degranulating actions took less than 5 minutes to become maximal, were dependent upon the presence of cytochalasin B, and required intact pathways of arachidonic acid metabolism, as judged by the inhibitory actions of arachidonate antimetabolites. Neither lipid required more than trace concentrations of extracellular calcium to effect optimal amounts of enzyme release. In one or more of these regards the two lipids differed strikingly from other neutrophil-degranulating agents such as A23187, N-formyl-methionyl-leucyl phenylalanine, and phorbol myristate acetate. The mechanism of action of the two lipid stimuli, therefore, appear to differ somewhat from the mechanisms of the other three stimuli. PMID- 6274202 TI - Up-regulation of hepatic prostaglandin E receptors in vivo induced by prostaglandin synthesis inhibitors. AB - Up-regulation in vivo of liver plasma membrane receptors for prostaglandin E (PGE) was studied in Sprague-Dawley rats using the prostaglandin synthesis inhibitors, acetylsalicylic acid (ASA) and indomethacin (Indo). Following 4 days of treatment with ASA, the concentration of receptors and the affinity for binding were both significantly increased (Ro - +37%, KA = +62%). Following 4 days of treatment with Indo, the number of receptors was increased but the binding-site affinity was decreased (Ro = +40%, KA = -71%). Animals were then examined after treatment with either ASA or Indo for 1 day, a time when there was no significant decrease in PGE. After 1 day of treatment, the opposite changes in binding-site affinity were again observed, but there were no changes in the number of receptors with either drug, suggesting that the changes in affinity resulted from non-prostaglandin-related effects of the drugs. To ascertain the physiologic consequences of up-regulation, adenylate cyclase activity was measured in control and up-regulated membranes. There were no significant changes in basal or in PGE-stimulated adenylate cyclase activity. These data demonstrate that decreased endogenous PGE causes up-regulation of PGE receptors, but that this is not accompanied by increased adenylate cyclase activity. These data may indicate that PGE-stimulated adenylate cyclase operates maximally under normal receptor concentrations and that therefore its activity cannot be increased by regulatory changes in receptor density. PMID- 6274201 TI - Anaplastic human gliomas grown in athymic mice. Morphology and glial fibrillary acidic protein expression. AB - The morphologic and biochemical characteristics of human surgical biopsy specimens taken from 17 patients with anaplastic human gliomas and of athymic mouse-grown tumors derived from them were examined. Fourteen were categorized as glioblastoma multiforme, one as an anaplastic astrocytoma, one as a recurrent glioblastoma multiforme, and one as a gliosarcoma. Fifteen of 17 tumors stained positively immunohistochemically for glial fibrillary acidic protein (GFAP), a glial-specific marker. When portions of the 17 surgical biopsy specimens were injected into the flank subcutaneous space of athymic mice, 16 produced tumors; different portions of a single biopsy specimen were used to establish three separate tumor lines; in toto, 18 tumor lines were established. Mouse-borne tumors contained various proportions of fibrillary and protoplasmic astrocytes, gemistocytes, small anaplastic cells, and multinucleated giant cells. Some were more homogeneous than the human tumors from which they were derived, while others contained a mixed population similar to that of the original biopsy specimen. Of these initial 18 tumors, 16 were stained for GFAP and 14 contained from fewer than 5% to almost 100% GFAP-expressing cells. Ten of the tumor lines were studied in serial passage, several demonstrating increased cellularity with increased passage. GFAP expression was followed through serial passage, and 7 of 10 tumor lines continued to express it, often in reduced amounts, and 2 of 10 ceased expression, one (the gliosarcoma) never having expressed it. These data demonstrate that while athymic mouse-borne human anaplastic gliomas retained some features of the human tumors from which they were derived, they varied from one another morphologically. These mouse-borne tumors also continued to evolve, often changing their levels of GFAP and demonstrating increased cellularity with passage. PMID- 6274203 TI - Action of thyrotropin-releasing hormone on mammotrophs and thyrotrophs. AB - Anterior pituitary cells from 15-day female rats were separated by unit gravity sedimentation into four populations (designated regions I-IV) based on the profile of cell distribution and the resulting content of radioimmunoassayable (RIA) hormones. The cells in regions II and IV released thyrotropin (TSH) in response to thyrotropin-releasing hormone (TRH, 5 ng/ml); however, those in region IV released only approximately 5% of their RIA content, whereas those in region II released approximately 26% in response to the same stimulus. Concomitant elevation of cAMP and of cGMP occurred in region II cells but only cGMP was elevated in region IV cells. Mammotrophs were localized in region III. They responded to TRH by releasing prolactin (PRL) and exhibiting increased cAMP content. These data provide support for the existence of two functionally distinct populations of thyrotrophs in 15-day-old female rats. The data also imply that cAMP is involved in TRH induced PRL release, whereas cGMP is involved in TRH-induced TSH release. PMID- 6274204 TI - Hormonal induction of Na-K-ATPase in developing proximal tubular cells. AB - The outer 150-micrometers layer of the renal cortex of the rat consists mainly of proximal tubules (PT). In this layer Na-K-ATPase (mumol Pi . mg protein-1 . h-1) increases from 3.45 +/- 0.19 (SE) in 10-day-old rats (R10) to 5.90 +/- 0.28 in 20 day-old rats (R20) to 9.52 +/- 0.42 in 40-day-old rats (R40). Betamethasone in pharmacologic doses increases Na-K-ATPase in R10, R20, and R40 to 11.5 +/- 1.19, 13.4 +/- 0.64, and 13.3 +/- 0.60, respectively. Estrogen in pharmacologic doses increases Na-K-ATPase to 9.1 +/- 0.46 in R20 and decreases Na-K-ATPase to 7.6 +/- 0.39 in R40. Aldosterone in a dose of 10 micrograms/100 g BW increases Na-K ATPase to 8.7 +/- 0.26 in R20; in a dose of 40 micrograms/100 g BW it increases Na-K-ATPase to 6.9 +/- 0.35 in R10. However, aldosterone in a dose of 80 micrograms/100 g BW is needed to cause an increase in R40 to 13.4 +/- 0.37. Treatment with canrenone from the 10th to the 20th day increases Na-K-ATPase. It is concluded that the immature PT cells are particularly sensitive to hormone induction of Na-K-ATPase and that the sensitivity is maximal during fairly late stages of cellular differentiation. Moreover, the inductive effect is probably mediated via glucocorticoid receptors. PMID- 6274206 TI - Prostaglandin E2-binding sites and cAMP production in porcine fundic mucosa. AB - Biologically active [3H]prostaglandin E2 (PGE2) bound rapidly and specifically to membrane fractions from hog fundic mucosa. Optimal binding occurred in the 30,000 g membrane preparation at 37 degrees C (pH 5.0). Scatchard analysis of specific PgE2 binding revealed the presence of a heterogeneous population of binding sites with Kd values and binding site concentrations of approximately 1 X 10(-9) M and 1 fmol/mg prot and 2 X 10(-8) M and 20 fmol/mg prot, respectively. Specific binding was inhibited by the following agents in descending order of potency: PGE1, PGA2, PGD2, 6-keto-PGF1 alpha, and thromboxane B2. Trypsin treatment or boiling reduced or abolished specific PGE2 binding. PGE2 stimulated cAMP formation in the 2,500-g fraction, with an approximate Km of 1 X 10(-6) M, but stimulation of adenylate cyclase activity by PG was not evident in the 16,000-g or 30,000-g tissue preparations. These results suggest that a specific PGE2 binding site exists in the 16,000-g and 30,000-g fractions of porcine fundic mucosa, although an increase in cAMP-forming capacity could not b of 1 X 10(-6) M, but stimulation of adenylate cyclase activity by PG was not evident in the 16,000-g or 30,000-g tissue preparations. These results suggest that a specific PGE2-binding site exists in the 16,000-g and 30,000-g fractions of porcine fundic mucosa, although an increase in cAMP-forming capacity could not b of 1 X 10(-6) M, but stimulation of adenylate cyclase activity by PG was not evident in the 16,000-g or 30,000-g tissue preparations. These results suggest that a specific PGE2-binding site exists in the 16,000-g and 30,000-g fractions of porcine fundic mucosa, although an increase in cAMP-forming capacity could not be localized in these fractions in vitro. PMID- 6274205 TI - Regulation of the PTH-receptor-cyclase system of canine kidney: effects of calcium, magnesium, and guanine nucleotides. AB - These studies examine the interactions of Ca2+, Mg2+, and guanine nucleotides with PTH binding to renal receptors and activation of adenylate cyclase in a purified preparation of basolateral renal cortical membranes. Inhibition of adenylate cyclase activity by increasing concentrations of CA2+ was more pronounced in the presence of PTH than on basal activity. This effect was not explained by an effect of Ca2+ on equilibrium PTH binding or binding affinity. The presence of PTH also decreased the requirements of the enzyme for Mg2+, consistent with the competitive nature of Ca2+ and Mg2+ interaction. Guanine nucleotides produced similar changes in the metal requirements for enzyme activity. PTH and guanine nucleotides, however, were not additive in this regard in spite of marked synergism in total enzyme activity. Additional studies were performed to further define the effects of guanine nucleotides on the receptor cyclase system. GTP and Gpp(NH)p caused a dose-dependent decline in equilibrium PTH binding and a small increase in half-maximal displacement. PTH-stimulated adenylate cyclase activity was markedly increased by either nucleotide, and the half-maximal activity was decreased (Gpp(NH)p greater than GTP). This was partly explained by a marked effect of guanine nucleotides on the dissociation rate of receptor-bound PTH. These data suggest that Ca2+ inhibition of PTH-stimulated adenylate cyclase activity is not due to a decreased binding of PTH but to an effect of PTH on the metal requirements for adenylate cyclase activation at the nucleotide regulatory site. PMID- 6274207 TI - Ca2+ mobilization in blood platelets as visualized by chlortetracycline fluorescence. AB - Binding of intracellular Ca2+ was measured in intact human blood platelets using the fluorescent Ca2+ probe, chlortetracycline, and a photon-counting microspectrofluorometer. Low doses of epinephrine, A23187, or prostaglandin endoperoxide analog U46619 induced a release of intraplatelet membrane-bound Ca2+. When platelet transmembrane Ca2+ flux was blocked by verapamil or ethylenediaminetetraacetic acid (EDTA), Ca2+ mobilization in response to epinephrine was inhibited, whereas A23187- or U46619-induced Ca2+ release was unchanged. When indomethacin was used to inhibit cyclo-oxygenase activity, Ca2+ mobilization in response to epinephrine or U46619 was partially blocked, whereas Ca2+ release in response to A23187 was unaltered. The relationship between platelet cyclic adenosine 3',5'-monophosphate (cAMP) and intraplatelet Ca2+ binding was also investigated. Prostaglandin E1 or prostacyclin was found to markedly elevate cAMP as well as enhance platelet Ca2+ binding. These effects were augmented by inhibition of phosphodiesterase activity using RO201724. The relationship between cAMP and Ca2+ binding was linear in the range of 10-60 pmoles cAmP/ml platelet-rich plasma. In addition the increase in cAMP stimulated by prostaglandin E1 or prostacyclin reduced the ability of epinephrine, A23187, or U46619 to induce intraplatelet Ca2+ mobilization. PMID- 6274208 TI - Waterborne gastroenteritis due to the Norwalk agent: clinical and epidemiologic investigation. AB - An outbreak of gastroenteritis occurred at a Pennsylvania summer camp in July 1978. Symptoms included abdominal pain (81 per cent), nausea (72 per cent), and vomiting (53 per cent); upper respiratory infection symptoms occurred in 35 per cent of the campers. Illness was associated with consumption of five or more glasses of water or water-containing beverages. Stool cultures from affected persons were negative for bacterial pathogens; however, a fourfold or greater rise to the Norwalk agent was demonstrated in serum samples of three of three ill persons tested and in none of eight controls (p < .02). Campers ill during the first session who were also present during the second session did not become ill during the second session (p < .001). (Am J Public Health 1982; 72:72-74.) PMID- 6274209 TI - Intraoperative biochemical localization of insulinomas by quick radioimmunoassay. AB - In patients with organic hyperinsulinism, intraoperative determination of insulin concentrations in veins draining the pancreas might help localize the tumor. We have developed a radioimmunoassay measuring insulin within 30 minutes. At operation, blood samples are taken at various sites in the splenic, superior mesenteric and portal veins for insulin determinations. In all six patients, the highest insulin concentrations correlated with the site of the tumor. Also, small insulinomas less than 1 cm in diameter could be found. In one patient the tumor appeared to secrete mainly proinsulin. By this procedure we hope to avoid blind distal resection of the pancreas and possibly invasive preoperative diagnostic procedures. PMID- 6274210 TI - [Hormonal status characteristics of women with rheumatic heart defects in the late stages of pregnancy]. PMID- 6274211 TI - [Immunologic disorders in trophoblastic disease]. PMID- 6274212 TI - [Possible role of herpes simplex virus type 2 in the etiology of cervical cancer]. PMID- 6274213 TI - [2 cases of trophoblastic tumor of the fallopian tube]. PMID- 6274214 TI - Prognostic factors of infantile spasms from the etiological viewpoint. AB - We investigated the prognostic factors for mental and physical development and seizure control by dividing the subjects into various etiologic groups in 200 patients with infantile spasms, all of whom (except 48 who died) were aged six years or older. The results were as follows: 1) Intermediate (4-12 mos) onset was found to be a favorable prognostic factor for seizure control in cryptogenic cases, although there was no relation between the age of onset and prognosis in other etiologic groups. 2) There was a significant correlation between the treatment lag and long-term prognosis for mental and physical development only in cryptogenic cases. A short treatment lag (0-2 mos) was associated with a good prognosis. All cryptogenic patients who had no relapse after ACTH therapy developed normally, although in symptomatic cases, there was no correlation between the relapse and the outcome. PMID- 6274215 TI - Peripheral neuropathy in children on long-term phenytoin therapy. AB - Nerve conduction studies of the ulnar, median, posterior tibial, peroneal and sural nerves were performed in 21 epileptic children aged 6 to 17 years on long term phenytoin therapy. Auditory brain stem evoked responses were obtained in 16 patients to evaluate the effect of phenytoin on central nervous system synapses. Of the 21 patients examined, 15 (71.4%) showed abnormal findings. The most frequent abnormality was slowed motor conduction velocity of the ulnar nerve (33.3%) and posterior tibial nerve (23.8%), followed by slowed sensory conduction velocity of the sural nerve (20%), lowered H/M ratio (14.3%), slowed motor conduction velocity of the peroneal nerve (14.3%) and of the median nerve (14.2%). A significant correlation was noted between the total dosage and duration of therapy with PHT and the reduction of motor conduction velocity in the posterior tibial nerve. Auditory brain stem evoked responses showed no significant differences in each peak latency between the patients and the normal control group. The study indicates that long-term phenytoin therapy can cause latent impairment of peripheral nerve function in children with no clinical evidence of peripheral neuropathy. PMID- 6274216 TI - Peripheral neuropathy in Marinesco-Sjogren syndrome. AB - A case of Marinesco-Sjogren syndrome manifesting cataract, short stature, psychomotor retardation, hypogonadism, and progressive ataxia was described. The result of electrophysiological study reflected the presence of peripheral nerve involvement, which was clearly confirmed by sural nerve biopsy. The conspicuous abnormality observed in the peripheral nerve was the presence of segmental demyelination. The process of axonal degeneration was not remarkable. Results confirmed that Marinesco-Sjogren syndrome is a disorder which may involve both central and peripheral nervous system. PMID- 6274217 TI - Dialyzable leukocyte extract stimulates cAMP in T gamma lymphocytes. AB - Dialyzable leukocyte extract (DLE) from human blood leukocytes gave a 3- to 8 fold increase in the cAMP concentration after addition to purified lymphocytes. No significant change in cGMP was observed. When purified lymphocytes were separated into T cells and non-T cells by an E-AET rosette technique the cAMP response was most pronounced in T cells. When further separating T lymphocytes into T cells with Fc receptors for IgG and T cells with Fc receptors for IgM (T gamma and T mu lymphocytes), the increase in cAMP following stimulation was specifically conferred to T gamma lymphocytes. Our findings may support the concept that DLE activates T gamma lymphocytes, leading to an increase in suppressor cell function. PMID- 6274218 TI - Molecular mechanisms of general anaesthesia. AB - This review aims to give a balanced view of the various mechanisms which have been proposed to explain the phenomenon of general anaesthesia on both a molecular and whole animal level. An attempt is made to interrelate these and produce one cohesive model. PMID- 6274219 TI - Microfingerprints of proteins through labeling acetylation of their proteolytic peptides. PMID- 6274220 TI - The simultaneous determination of the products of estrogen 2- and 4-hydroxylase action; the use of high-performance liquid chromatography with electrochemical detection. PMID- 6274221 TI - Spectrophotometric assay of NADase-catalyzed reactions. PMID- 6274222 TI - NADH/NADPH--cytochrome c reductase assay: interference by nonenzymatic cytochrome c reducing activity in plant extracts. PMID- 6274223 TI - Quantitative measurement of protease activities in slab polyacrylamide gel electrophoretograms. PMID- 6274224 TI - Detection of NAD(P)H--rubredoxin oxidoreductases in Clostridia. PMID- 6274225 TI - A method for quantitation of protein in the presence of Percoll. PMID- 6274226 TI - ESR study of intercalation: quantitative evaluation of drug-DNA binding through competition with a spin-labeled 9-aminoacridine. PMID- 6274227 TI - An improved method for determining the actin filament content of nonmuscle cells by the DNase I inhibition assay. PMID- 6274228 TI - Preparation, properties, and uses of two fluorogenic substrates for the detection of 5'-(venom) and 3'-(spleen) nucleotide phosphodiesterases. PMID- 6274230 TI - A rapid method for the fractionation of isolated hepatocytes. PMID- 6274229 TI - A fluorogenic method for the demonstration of human serum 5'-nucleotide phosphodiesterase isozymes after polyacrylamide gel electrophoresis. PMID- 6274231 TI - One-step molybdate method for rapid determination of inorganic phosphate in the presence of protein. PMID- 6274233 TI - Comparison of equipotent doses of non-depolarizing muscle relaxants in children. AB - The neuromuscular effects of equipotent doses of non-depolarizing muscle relaxants used for endotracheal intubation were studied in 27 children anesthetized with thiopental, nitrous oxide/oxygen, and narcotic. Equipotent doses of d-tubocurarine (0.8 mg/kg), metocurine (0.5 mg/kg), and pancuronium (0.13 mg/kg) were used. At these doses conditions for intubation were satisfactory in all children and the twitch was completely abolished in 26 of the 27 patients. The twitch height recovered to 5% of control values in 54 +/- 6 minutes, and 29 +/- 2 minutes later recovered to 25% of control values. The time from injection of the drug to maximum effect, the conditions for intubation, and recovery times among the three drugs were not significantly different. Train-of four values correlated (p less than 0.001) with the twitch heights. When the twitch height at 0.1 Hz was 21% of control values, only three contractions were detected following train-of-stimulation. At 14% of control twitch height, two contractions were detected; at 7% of control twitch height, one contraction was detected. The most frequent reason for administering an incremental dose of relaxant was the beginning of respiratory movements, corresponding to a twitch height of 24% of control values (train-of-four 3%). The second most common reason was unsatisfactory abdominal wall relaxation (twitch height 31% of control values, train-of-four 11%. PMID- 6274232 TI - Evaluation of PS beta 1G and other tumor markers in germ cell tumors of the testis. AB - Pregnancy Specific beta 1 Glycoprotein (PS beta 1G), human choriogonadotropin (hCG), alpha foeto protein (AFP) and Carcino Embryonic Antigen (CEA) were assayed in 58 patients with active germ cell tumors and 20 patients in complete clinical and radiological remission. The markers were negative in all the cases in remission as well as in one case of pure teratoma. The positivity rate of the four markers together is 75%, which is better than for any marker alone. All the markers except AFP seem ubiquitous with respect to the histological classification. The highest positivity rate is obtained for hCG in all the histological types. PS beta 1G is related to hCG but its positivity rate is quite lower. AFP is mainly related to embryonal carcinoma but can be associated with any type of tumor except seminoma. CEA is less frequently positive, always associated with other markers and seems therefore useless for the diagnosis of germ cell tumors. PMID- 6274234 TI - Absorbable suture materials for vascular anastomoses. Tensile strength and axial pressure studies using polyglycolic acid sutures. AB - This study attempted to evaluate how vascular healing itself is sufficient enough to provide adequate strength to autogenous vein grafts in small arteries without the use of nonabsorbable sutures. The absorbable suture, Dexon, was used in small canine femoral vein graft anastomoses over a two-month trial period with nonabsorbable Prolene as a control. Tensile strength and resistance to axial leak were measured. Results show that Dexon had 93 per cent of the tensile strength of Prolene immediately; 76 per cent at 14 days, 111 per cent at 21 days, and 110 per cent at 30 days, and 124 per cent at 60 days as an average for all suture lines. The per cent mean resistance to leakage that Dexon had compared to Prolene for all suture lines was 102 per cent at seven days, 97 per cent at 14 days, 107 per cent at 21 days, and 97 per cent at 30 days. There is no evidence of aneurysms, suture line disruptions, dilations, or infections of any anastomoses. Our ultimate conclusion is that the integrity of small autogenous vein graft is dependent on healing rather than on the permanent strength of the suture material and that Dexon maintains vascular integrity long enough to permit this healing to occur. PMID- 6274236 TI - Herpes zoster in small-cell carcinoma of the lung. PMID- 6274235 TI - Nabilone and cancer chemotherapy. PMID- 6274237 TI - [The use of electrical transcutaneous nerve stimulations in chronic pain. Method and preliminary results in 28 cases (author's transl)]. AB - The analgesic effect of conventional transcutaneous nerve stimulation has been studied in 29 patients, all having a chronic pain caused by peripheral neurological disease. As already reported in the literature half of patients were improved on a short-term basis. Long-term improvement was observed mainly in patients with traumatic nerve lesions; such cases, thus appear to be the best indication for this method. It should be emphasized that whatever the etiology, the delay between the onset of pain and the beginning of transcutaneous stimulations was a critical factor. Nine out of ten patients suffering for less than one year were satisfactorily improved. This suggests that transcutaneous stimulations should be used as early as possible after the onset of neurological pain. PMID- 6274238 TI - [Hypersensitivity of the beta-adrenergic receptors in patients considered as spasmophilic. Results of two stimulation trials with isoprenaline in 105 patients (author's transl)]. AB - Two stimulation trials with isoprenaline were performed in two groups of 50 and 55 patients considered as spasmophilic. The first trial was to check the sensitivity of the beta 1 type cardiac receptors : the average isoprenaline hydrochloride dose raising by 50 p. 100 the rest cardiac rate was equal to 0.147 +/- 0.09 micrograms/kg for 0.23 +/- 0.09 micrograms/kg in the control group. A second trial allowed to stabilize the cardiac rate between + 40 and + 50 p. 100 of its initial value : the symptoms, reappearing during stimulation, under the influence of isoprenaline, were studied. In 43 out of the 50 investigated patients the usual symptoms reappeared, particularly the acrocontractural attacks which were observed then in 46 p. 100 of cases, and sometimes an acute tetanic attack with opisthotonus. All these signs which are characteristic of "the spasmophilic state" seem to relate to vegetative regulation in which the beta adrenergic component would show the most important : hypersensitivity of the beta receptors might be the common physiological base. PMID- 6274239 TI - Subchronic ocular and systemic toxicity of topically applied delta 9 tetrahydrocannabinol. PMID- 6274240 TI - [Mucinous adenocarcinoma of the ovary with pseudosarcomatous nodules (presentation of a case)]. PMID- 6274241 TI - [Radionuclide study of thyroid function in pediatrics, using 99mTc, 123I or 131I: 150 case-reports (author's transl)]. PMID- 6274242 TI - Mixed mesodermal tumour of the fallopian tube. A case report. PMID- 6274243 TI - [Plasma DHA, DHAS and cortisol levels after injection of beta 1-24 corticotrophin beta 1-24 ACTH in normal adult women (author's transl)]. AB - Plasma DHA, DHAS and cortisol levels were measured by R.I.A. before and after intravenous and intramuscular injection of beta 1-24 ACTH. In 7 normal adult women, plasma cortisol and DHA levels rose sharply in response to beta 1-24 ACTH (0,25 mg) i. v injection; in opposition, plasma DHAS levels did not change significantly 30 and 120 minutes after the injection. In 9 normal adults women, the i. m injection of Zn beta 1-24 ACTH (1 mg) caused a significant increase in both plasma cortisol, DHA and DHAS levels 8 and 24 hours after the injection. PMID- 6274244 TI - [Acromegaly and sleep apnea syndrome (author's transl)]. AB - Acromegaly associated with a Sleep Apnea Syndrome has but exceptionally been reported. Polygraphic recordings of sleep have been carried out in parallel with the determination of pituitary hormonal secretions, during the nycthemeral period before and after surgical treatment of the adenoma. There appears a Sleep Apnea Syndrome of the predominant obstructive type; the Apnea index is: 57 (N less than or equal to 4); the hypnogram is considerably jagged, with more than a thousand wakings and changes in the sleep stages, due to a great number of apneas. The deep slow sleep never occurs: no stages 3 and 4. The physiological peak of G.H. secreted in the beginning of the deep slow sleep thus does not appear in the Sleep Apnea Syndrome. The existence of a "false negative" criteria of a cured Acromegaly must be taken into consideration. The Sleep Apnea Syndrome must be differentiated from Narcolepsy and the usual Pickwickian syndrome. The Sleep Apnea Syndrome and Acromegaly seem to be two separate diseases, each one evolving independently. The cure of Acromegaly has not led to the cure of the Sleep Apnea Syndrome and the latter has not prevented the clinical and biological cure of Acromegaly. This may be an argument in favor of the independence of Acromegaly towards some hypothalamic structures. PMID- 6274245 TI - [Influence of nutritional factors on the enhancement of renal compensatory hypertrophy by hyperadrenocorticism in the rat (author's transl)]. AB - In the rat compensatory hypertrophy (RCH) was enhanced by hyperadrenocorticism induced by the administration of a long acting ACTH at a dose of 18 Y/100 g body weight/d. for 7 d. after uninephrectomy (UN). In the present experiments we compared the differences delta between the weight, the content in protein, RNA and DNA of the left solitary kidney and the same determinations done on the right kidney excised at UN 7 d. earlier. The rats drank freely a isotonic solution of NaCl (G1) or KCl (G2) or glucose (G3, G4). The rats of group G1, G2 and G3 received a standard solid food; the G4 rats ate a K poor diet. About half of the animals were treated with ACTH. RCH occurred in all the rats even when they lost body weight. The pain in weight of the solitary kidney was enhanced in all the rats treated with ACTH but not in the G2 rats loaded with KCl. This renotrophic action of hyperadrenocorticism was most prominent in the K depleted G4 rats. The protein/DNA ratio, a marker of cellular hypertrophy, was increased by hyperadrenocorticism in the G1 and G2 rats drinking respectively the NaCl or the KCl solutions. This ratio did not change in the ACTH treated G3 and G4 rats drinking the glucose solution suggesting that, in this experimental condition, cellular hyperplasia and hypertrophy occurred at the same extent. These experiments suggest that, in the uninephrectomized rat, the renotrophic action of ACTH is modulated by nutritional factors. The enhancement of RCH by ACTH may be related to hyperglycemia, hyperinsulinism or altered handling of Na+ and K+ by the nephron. PMID- 6274246 TI - Pituitary adrenal axis during transsphenoidal pituitary adenomectomy. PMID- 6274248 TI - Diphenylhydantoin inhibits calcium fluxes and insulin release in pancreatic islets. PMID- 6274249 TI - Effects of diabetes on somatogenic and lactogenic receptors in rat liver. PMID- 6274247 TI - Changes in cortisol metabolism in various physiological and pathological situations. PMID- 6274250 TI - Biochemical characterization of the genetic variants of human phosphoglycolate phosphatase (PGP). AB - Phosphoglycolate phosphatase (PGP) exhibits a wide range of activities in normal human red cells. Analysis of blood from 57 individuals of known PGP phenotype revealed no correlation between enzyme activity, electrophoretic phenotype or 2,3 DPG concentration. Neither was there evidence of variation in Km, heat stability, pH optimum or molecular size among the various electrophoretic phenotypes. However, human red cell PGP exhibits high (0.24 mM) and low (0.05 mM) Km values for phosphoglycolate irrespective of phenotype and this requires further analysis. PMID- 6274251 TI - Linkage relationship of the loci for Anderson--Fabry disease and the Xg blood groups. AB - Information provided by the follow-up of a family in which Anderson--Fabry disease segregates, together with a family reported by Ropers et al. (1977), negates the previous evidence of a probable linkage between alpha GALA and Xg. PMID- 6274252 TI - Endorphins in experimental spinal injury: therapeutic effect of naloxone. AB - An experimental model of cat spinal injury was used to investigate the hypotheses that endorphins are involved in the pathophysiology of spinal cord injury and that the opiate antagonist naloxone, by blocking the effects of endorphins, may improve physiological and neurological recovery. Experimental trauma to the cervical spine caused a reduction in blood pressure and spinal cord blood flow (SCBF) associated with increased plasma levels of beta-endorphin-like immunoreactivity; both blood pressure and SCBF were significantly improved by naloxone treatment. Naloxone-treated animals also showed less prominent spinal cord abnormalities and significantly improved neurological recovery compared with saline controls. Moreover, both the pathological and clinical findings correlated significantly with the blood pressure changes following treatment. The data support a pathophysiological role for endorphins in experimental spinal cord injury and suggest that opiate antagonists might improve the treatment of spinal trauma in humans. PMID- 6274253 TI - ACTH-induced cortisol production in multiple sclerosis. AB - We measured the total daily urinary cortisol produced by each of 8 patients with multiple sclerosis during intravenous therapy with adrenocorticotropic hormone (ACTH). Results indicated marked daily variation from patient to patient as well as in the same patient, suggesting that ACTH infusions do not result in reliable, consistent, high-level production of cortisol. We attempted to express the quantity of cortisol produced as an equivalent amount or oral prednisone. This comparison is difficult to make; but, given certain assumptions, the figures suggest that 40 U or 80 U ACTH infusions do not elicit the quantities of steroid generally considered to be indicated for the therapy of major allergic, autoimmune, or neoplastic diseases. PMID- 6274255 TI - GABA receptors in alcoholism. PMID- 6274254 TI - A family affected with intestinal polyposis and gliomas. AB - The third affected family with Turcot syndrome is presented and the problems associated with the occurrence of familial aggregates of cancer are discussed. Turcot syndrome represents the unique and discrete occurrence of polyposis coli with glioblastoma multiforme, medulloblastoma, or both. PMID- 6274256 TI - Mechanism of poliovirus inactivation by bromine chloride. AB - The mechanism of poliovirus inactivation by BrCl was determined by exposing poliovirus to various concentrations of BrCl and correlating the loss of virus infectivity with structural changes of the virus. Concentrations of 0.3 to 5 mg of BrCl per liter resulted in 95% to total inactivation of poliovirus. However, the inactivated virus retained structural integrity, as determined by buoyant density measurements of poliovirus labeled with radioactivity. However, at concentrations of 10 to 20 mg of BrCl per liter, total inactivation of poliovirus was associated with the degradation of the structural integrity of the virus. Since infectious ribonucleic acid at similar concentrations could be recovered from untreated poliovirus and poliovirus treated with 0.3 mg of BrCl per liter, it was concluded that BrCl as HOBr or bromamines inactivates poliovirus by reacting with the protein coat of the virus. Moreover, this inactivating reaction does not result in the degradation of the structure of the virion, nor does it affect the biological activity of the internal ribonucleic acid of the virus. PMID- 6274257 TI - Positively charged filters for virus recovery from wastewater treatment plant effluents. AB - Positively charged Zeta Plus filters were used to concentrate enteroviruses from 19 liters of effluent from activated sludge units. Neither the addition of salts nor the acidification of the effluent was required for adsorption of viruses to the filters. Viruses adsorbed to the filters were eluted by treating the filters with a solution of 4 M urea buffered at pH 9 with 0.05 M lysine. Eluted viruses were concentrated into final volumes of 1 to 2 ml by using a two-step concentration procedure that employed inorganic and organic flocculation. Approximately 50% of the viruses added to effluents could be recovered in the final sample. The procedure was used to monitor effluents from activated sludge units at two wastewater treatment plants for the presence of enteroviruses. PMID- 6274258 TI - Recovery of indigenous enteroviruses from raw and digested sewage sludges. AB - We examined different types of raw sewage sludge treatment, including consolidation, anaerobic mesophilic digestion with subsequent consolidation, and aerobic-thermophilic digestion. Of these, the most efficient reduction in infectious virus titer was achieved by mesophilic digestion with subsequent consolidation, although a pilot-scale aerobic-thermophilic digester was extremely time effective, producing sludges with similarly low virus titers in a small fraction of the time. Although none of the treatments examined consistently produced a sludge with undetectable virus levels, mesophilic digestion alone was found to be particularly unreliable in reducing the levels of infectious virus present in the raw sludge. PMID- 6274259 TI - Poliovirus adsorption by 34 minerals and soils. AB - The adsorption of radiolabeled infectious poliovirus type 2 by 34 well-defined soils and mineral substrates was analyzed in a synthetic freshwater medium containing 1 mM CaCl(2) and 1.25 mM NaHCO(3) at pH 7. In a model system, adsorption of poliovirus by Ottawa sand was rapid and reached equilibrium within 1 h at 4 degrees C. Near saturation, the adsorption could be described by the Langmuir equation; the apparent surface saturation was 2.5 x 10(6) plaque-forming units of poliovirus per mg of Ottawa sand. At low surface coverage, adsorption was described by the Freundlich equation. The soils and minerals used ranged from acidic to basic and from high in organic content to organic free. The available negative surface charge on each substrate was measured by the adsorption of a cationic polyelectrolyte, polydiallyldimethylammonium chloride. Most of the substrates adsorbed more than 95% of the virus. In general, soils, in comparison with minerals, were weak adsorbents. Among the soils, muck and Genesee silt loam were the poorest adsorbents; among the minerals, montmorillonite, glauconite, and bituminous shale were the least effective. The most effective adsorbents were magnetite sand and hematite, which are predominantly oxides of iron. Correlation coefficients for substrate properties and virus adsorption revealed that the elemental composition of the adsorbents had little effect on poliovirus uptake. Substrate surface area and pH, by themselves, were not significantly correlated with poliovirus uptake. A strong negative correlation was found between poliovirus adsorption and both the contents of organic matter and the available negative surface charge on the substrates as determined by their capacities for adsorbing the cationic polyelectrolyte, polydiallyldimethylammonium chloride. PMID- 6274261 TI - Cutaneous malignant mixed tumor. AB - Two cases of cutaneous malignant mixed tumor occurred in a 70-year-old man and a 69-year-old man. Nine previously described patients with cutaneous malignant mixed tumor resembled the two patients in this report, both clinically and histologically. All cases with adequate long-term follow-up evaluations were characterized by recurrence or metastasis. In contrast to benign mixed tumors, these tumors tend to occur on the extremities, are of larger size, and may exhibit rapid growth. Histologically, cellular atypism, increased cellularity, increased mitotic rate, invasion of surrounding tissue, and necrosis may be identified. PMID- 6274260 TI - Influence of pH and electrolyte composition on adsorption of poliovirus by soils and minerals. AB - The pH and the nature an concentration of simple electrolytes influenced the interaction of poliovirus type 2 with three soils, a sand, and a clay mineral. In electrolytes above pH 9 the virus was not adsorbed extensively to the substrates, but below pH 7 almost all virus was bound. For each adsorbent there was a characteristic pH region of transition from strong to weak uptake. Differences between the soils in virus uptake were shown to parallel their pH-dependent mineral. In electrolytes above pH 9 the virus was not adsorbed extensively to the substrates, but below pH 7 almost all virus was bound. For each adsorbent there was a characteristic pH region of transition from strong to weak uptake. Differences between the soils in virus uptake were shown to parallel their pH dependent mineral. In electrolytes above pH 9 the virus was not adsorbed extensively to the substrates, but below pH 7 almost all virus was bound. For each adsorbent there was a characteristic pH region of transition from strong to weak uptake. Differences between the soils in virus uptake were shown to parallel their pH-dependent charge properties, as determined by whole-particle microelectrophoresis. Only when the pH was close to or above the critical region was uptake increased with electrolyte concentration. The transition region for all substrates was above pH 7.5 the isoelectric point of the virus. Thus, it appears that when both the virus and substrate are highly negative charged, repulsive electrostatic effects may exceed inherent attractive interactions, thereby inhibiting adsorption. PMID- 6274263 TI - Effect of cAMP, Mn2+, and phosphodiesterase inhibitors on human sperm motility. AB - The effects of cyclic adenosine monophosphate (cAMP), MnCl2, caffeine, and theophylline at concentrations of 10 mM on human sperm motility and forward migration in vitro were tested. cAMP was effective in activating human spermatozoal motility and forward migration after incubation with the spermatozoa for 3 hr or more and 5 hr, respectively, whereas caffeine and theophylline were effective in activating sperm motility after incubation with the spermatozoa for 1 hr or more. Caffeine and theophylline significantly activated sperm forward migration only after incubation with the sperm for 5 hr. MnCl2, a potent sperm adenylate cyclase activator, had no significant effect in improving human sperm motility and forward migration. PMID- 6274262 TI - A clinically relevant canine lung cancer model. AB - Research on early human lung cancer is difficult; we have sought a canine correlate. Regimens included endobronchial submucosal injections and topical focal applications of benzo[a]pyrene, nitrosomethylurea, dimethylbenzanthracene, and methylcholanthrene, singly or in combinations. Sustained-release discs were placed into lung parenchyma or sutured into major bronchi. Tracheal segments were isolated as cervical pedicle grafts. Gross and histological evolution was reproducible. Columnar and basal hyperplasia and squamous metaplasia were early changes. Atypia occurred within 6 weeks and was found in all dogs within 16 to 18 weeks. Invasive cancers occurred within 8 to 65 months. No tracheal graft developed cancer. Of 15 dogs with parenchymal sustained-release implants, 1 to date has developed cancer in 8 months. Four endobronchial regimens have produced 16 cancers in 56 lungs at risk for 18 to 65 months. No cancers developed in 23 lungs at risk from eight other regimens. Of 10 dogs at risk for unilateral endobronchial cancer, 5 have had cancer. Of 23 dogs with both lungs at risk, 9 developed cancer. We have shown focal carcinogenesis with well-defined pathogenesis and an extended preneoplastic period at predictable sites in a lung cancer model. PMID- 6274264 TI - Secondary hyperparathyroidism caused by oral contraceptives. AB - Secondary hyperparathyroidism developed on two separate occasions in a patient taking an oral contraceptive (OC) in a mixture of norethindrone and mestranol (Ortho-Novum 1/80). The probable cause of this condition is an estrogenic inhibition of the calcium mobilizing effect of parathyroid hormone (PTH) or vitamin D. The possible consequences of OC therapy in this patient and patients like her include those of milk hypocalcemia, moderate hypophosphatemia, or the neurotoxic actions of excessive levels of circulating PTH. One should also consider the possible role of OC therapy in patients with unexplained parathyroid hyperplasia and with tertiary hyperparathyroidism. PMID- 6274265 TI - Polymicrobial septicemia associated with rhabdomyolysis, myoglobinuria, and acute renal failure. AB - Myoglobinuria and renal failure resulting from bacterial infection have only rarely been reported. To our knowledge, we describe the first reported case of polymicrobial septicemia resulting in rhabdomyolysis and myoglobinuric renal failure. Renal failure secondary to myoglobinuria has an excellent prognosis; in our patient, recovery was complete. The frequency of rhabdomyolysis, myoglobinuria, and renal failure in septicemia is unknown and can only be determined by an increased awareness of this potential complication of septicemia. PMID- 6274266 TI - Application of a microwave tissue coagulator to hepatic surgery the hemostatic effects on spontaneous rupture of hepatoma and tumor necrosis. PMID- 6274267 TI - Platelet alpha 2-adrenergic receptors in major depressive disorder. Binding of tritiated clonidine before and after tricyclic antidepressant drug treatment. AB - The specific binding of tritiated (3H)-clonidine, an alpha 2-adrenergic receptor agonist, to platelet membranes was measured in normal subjects and in patients with major depressive disorder. The number of platelet alpha 2-adrenergic receptors from the depressed group was significantly higher than that found in platelets obtained from the control population. Treatment with tricyclic antidepressant drugs led to significant decreases in the number of platelet alpha 2-adrenergic receptors. These results support the hypothesis that the depressive syndrome is related to an alpha 2-adrenergic receptor supersensitivity and that the clinical effectiveness of tricyclic antidepressant drugs is associated with a decrease in the number of these receptors. PMID- 6274268 TI - Presynaptic adrenergic receptor sensitivity in depression. The effect of long term desipramine treatment. AB - Stimulation of presynaptic alpha 2-adrenergic receptors located on norepinephrine (NE)-containing cells in the brain decreases the firing rate and turnover of NE in these neurons. To assess whether abnormalities in the regulation of the NE system during desipramine hydrochloride treatment may be present in depressed patients, the effects of an alpha 2-agonist, clonidine hydrochloride, on plasma levels of the NE metabolite 3-methoxy-4-hydroxy/phenethyleneglycol (MHPG) and on blood pressure (BP) were evaluated in ten depressed patients before and during long-term desipramine treatment. Long-term desipramine treatment significantly attenuated the effects of clonidine on plasma MHPG level and BP, indicating that during desipramine treatment alpha 2-adrenergic receptors had become subsensitive. In addition, plasma MHPG levels were significantly reduced during long-term desipramine treatment. These findings are discussed in relation to the hypothesized therapeutic mechanism of action of desipramine and the hypotheses relating noradrenergic function and depression. PMID- 6274269 TI - Studies of the purification of tumor antigens: progress and problems. PMID- 6274270 TI - Is there any diagnostic relevance of human antibodies which react with mouse mammary tumor virus (MuMTV)? AB - Applying an indirect immunofluorescence assay with prolonged serum incubation on frozen-cut mouse mammary tumor slices, antibodies are detectable in human sera which react with viral inclusion bodies consisting of intracytoplasmic A particles (iAp). These iAp are known to represent intracellular entities of the mouse mammary tumor virus (MuMTV), mainly related to MuMTV core constituents. In 1,449 women studied, the antibody incidence is partially correlated with proliferating changes of the mammary gland. However, epidemiological data obtained thus far have clearly shown that the detection of antibody is not useful in breast cancer diagnosis (antibody incidence in breast cancer patients 22.8%, in lactating women 20.6%, in controls 5.9%). Studying antibody reagibility to particular iAp polypeptides by means of a radioimmunoprecipitation (RIP) technique, there has been preliminary evidence that human serum factors react with more than one mouse virus protein. The most prominent reactivity was seen to be directed to the 14,000 dalton protein (Ap14). The detection of serum antibody activity is considered as being of important indicative value in that it reflects the existence of MuMTV-related antigens in man. PMID- 6274271 TI - Metastatic adenocarcinoma of unknown primary site: diagnostic electron microscopy to determine the site of tumor origin. AB - Poorly differentiated malignant tumors and metastatic adenocarcinomas have been studied by electron microscopy. Rapid ultrastructural analysis allows the accurate diagnosis of the cell of tumor origin to be reported in conjunction with the routine surgical signout of paraffin sections. The identification of specialized intracellular structures is useful for determining the site of origin of metastatic tumors. Particularly useful in this material was the presence of lamellar (surfactant) bodies, typical of alveolar cell carcinoma of the lung, as well as the presence of apical terminal webs, diagnostic of gut epithelial cells. PMID- 6274272 TI - Congenital glioblastoma multiforme associated with congestive heart failure. AB - A case of congenital glioblastoma multiforme in a premature hydropic female infant was studied. Study and results of a literature review show that teratoma is the commonest tumor in that age group, and that the sex distribution in newborns showed a slight preponderance in females, in contrast with the preponderance of these neoplasms in males among children and adults. PMID- 6274273 TI - Salivary gland heterotopia: a unique cerebellopontine angle tumor. AB - An unusual maldevelopmental lesion, a salivary gland heterotopia of the right cerebellopontine angle, was studied. There was an associated primary adenoid cystic carcinoma. Possible modes of development and the relationship to salivary heterotopia of the middle ear are discussed. PMID- 6274274 TI - [Fructose. Physiopathology of its metabolism and critical review of the therapeutic uses]. PMID- 6274275 TI - Subacute combined degeneration, neutrophilic hypersegmentation, and the absence of anemia. A case report. PMID- 6274276 TI - Prevention of chemotherapy-induced nausea and vomiting in patients with cancer. PMID- 6274277 TI - [Malignant fibrous histiocytoma of the bone]. AB - The data of clinico-morphological study of 10 observations of malignant fibrous histiocytoma of the bone indicating its distinct nosological importance are presented. Clinico-morphological comparisons showed the typical variant of the morphological structure to have the worst prognosis. In two observations, the tumor was examined electron microscopically. All the 5 main types of cells were found: histiocyte- and fibroblast-like, giant multinuclear, xanthomatous, and poorly-differentiated mesenchymal. In one case, intranuclear virus-like inclusions of the herpesvirus type were found. Their presence in 4 main types of cells suggests their common histogenetic origin from poorly-differentiated mesenchymal cells. PMID- 6274278 TI - [1 of the variants of Wilms' tumor]. AB - A peculiar variant of Wilms' tumor was studied. Out of 77 cases of nephroblastoma 2 observations were selected in which the tumor consisted of loosely arranged oval and polygonal cells with clear nuclei. Occasional formed tubules were found in the tumor. The cells had anastomosing processes, few organoids, microfibrils. A nonepithelial nature of the tumor is assumed. A trend for primary metastasis of the tumor into bones was observed. Because of the difficulties and rare occurrence of such tumors, their histology and differential criteria are presented at the light and ultrastructural levels. Indication of this variant in the diagnosis may also suggest the possibility of metastatic affection of the skeleton. PMID- 6274279 TI - [Brain lesions in children with para-influenza]. AB - An analysis of 1274 autopsy protocols of children parainfluenza was revealed as the main disease in 73 and as a concomitant disease in 107 cases, not infrequently as a generalized infection. In 13 observations a detailed study of the brain was performed. In 4 cases the involvement of the brain consisted mainly in cushion-like outgrowth of ependymocytes and endotheliocytes of the blood vessels. In 1 case a comprehensive analysis revealed isolated meningoencephalitis induced by type 3 parainfluenza virus. Besides, hemodynamic disorders and the nonspecific changes of moderate intensity were found. In 8 children only these nonspecific changes were detected. PMID- 6274280 TI - [Structural characteristics of the tumor and the state of the surrounding tissue in middle-aged and elderly breast cancer patients]. AB - Investigations of mammary glands of 241 patients of 50-70 years of age or older suffering from carcinoma and having been is menopause for 10 years or more established that the main histological form of carcinoma was infiltrating carcinoma of glandular, solid, solid-scirrhous structure of various degrees of malignancy (I, II, III) (81.3%), irrespective of the age and duration of menopause. The main forms of carcinoma were observed in occasional cases, most frequently it was lobular invasive carcinoma (8.3%). In 65% of the patients carcinoma was detected in the presence of involutive processes and changes corresponding to nonproliferating mastopathy. In one-third of the patient carcinomas developed in the presence of proliferative processes, half of these patients showing transition from proliferation of the epithelium of ducts and lobules to cancer in situ and its invasion into the surrounding tissues of mammary glands. PMID- 6274281 TI - [Organ-specific ultrastructural cellular signs of cancerous breast tumors]. AB - The ultrastructural analysis of 40 cases of mammary carcinomas revealed two groups of cells: with ultrastructural organ specific features (differentiated cells) and without them (undifferentiated cells). Intracytoplasmic lumens, secretory granules (lipid, protein, and mixed lipid-protein), and myoepithelial cells are the main organ specific features of mammary carcinomas. The histospecific features showing the epithelial origin of the tumor cells include cell-to-cell contacts, ductules, intercytoplasmic lumens with microvilli, and fragments of basal lamina. These ultrastructural features of mammary carcinomas can facilitate the differential diagnosis of tumors of this organ. PMID- 6274282 TI - [Pigmented neuroectodermal tumor (melanoameloblastoma)]. AB - A pigmented tumor (melanoameloblastoma) in a 4-month-old infant localized in the upper jaw is described. The tumor consisted of cells with melanin granules in the cytoplasm. The detected neurosecretory granules and melanosomes in the parenchymal cells of the tumor suggest to be of neuroectodermal origin. PMID- 6274283 TI - Paraneoplastic encephalomyelitis and neuropathy. Report of a case. PMID- 6274284 TI - Cisplatin therapy for adenoid cystic carcinoma. AB - Adenoid cystic carcinoma of salivary gland origin is a relentless disease that produces a high rate of local recurrence, metastases, and death, regardless of the type of treatment. There have been few previous reports of effective chemotherapy. Cisplatin has been chosen for a trial in the treatment of patients with persistent, recurrent, or metastatic adenoid cystic carcinoma because of its demonstrated activity against a variety of tumor types, especially those with a low growth fraction. Ten patients have been included in a pilot study. Seventy percent of these patients have had a favorable subjective response and objective tumor regression. Further clinical trials are suggested. PMID- 6274285 TI - Salivary gland tumors of the lip. AB - The UCLA experience with minor salivary gland tumors of the lip is presented and contrasted with that of the literature. The incidence of benign to malignant tumors of the lip does not follow the inverse relationship stated in the axiom that the smaller the salivary gland the greater the probability that a developing tumor will be malignant. Benign tumors represent over 80% of all salivary gland tumors of the lip. There is no preponderant malignant tumor for the lip. Adenoid cystic carcinoma, mucoepidermoid carcinoma, and adenocarcinoma occur with almost equal frequency. Because of the indolent nature of these tumors, excellent survival rates can be achieved with wide local excision with few recurrences, if the tumors are adequately treated when first seen. PMID- 6274286 TI - Sweat gland tumors of the head and neck. AB - Surgeons performing soft-tissue surgery in the head and neck area occasionally will encounter other varieties of neoplasms, such as skin-appendage tumors, in addition to basal cell carcinoma and squamous cell carcinoma. Sweat gland tumors are the most common variety of the skin-appendage tumors and the most easily confused with basal cell carcinoma and squamous cell carcinoma. We report this case to illustrate this fact. The classification of sweat gland tumors and their suggested management are reviewed. PMID- 6274287 TI - The histogenesis of so-called mixed tumor evaluating from parotin binding. AB - We evaluated the histogenesis of so-called mixed tumors of the parotid gland by comparing the tumor cells with normal parotid gland cells. Using the indirect technique of the enzyme antibody method, the paraffin sections of human parotid glands, initially reacted with 0.1 mg/ml of parotin solution in PBS for 20 min at 37 degrees C, were treated for 20 min at 37 degrees C with anti-parotin subunit rabbit IgG (0.01 mg/ml) containing 5 mg/ml BSA, and then with HRPO-labeled anti rabbit gamma globulin antibodies (swine IgG). After a staining for enzyme by Karnovsky's method, DAB reactions were observed on the duct cells of human parotid glands in 12 of 13 cases. We surveyed the parotin binding as a marker of duct cell on the so-called mixed tumors of 11 cases. DAB reactions were recognized on the cells of myxoid portion, along with the polygonal cells of solid sheet, board cord, and ductular formation, but the cells of chondroid portion and the spindle-shaped cells forming trabecular structure did not. PMID- 6274288 TI - Bilateral breast cancer in a Chinese male. PMID- 6274289 TI - The transmission of two new Australian serotypes of bluetongue virus to sheep. PMID- 6274290 TI - Monospecific fluorescent antiserum for study of bovine viral diarrhoea virus. PMID- 6274291 TI - Outbreaks of egg-drop syndrome-1976 in Japan and its etiological agent. AB - A condition similar to egg-drop syndrome-1976 (EDS-76) occurred in 14 broiler breeding flocks in 2 farms in Japan from December 1978 to January 1980, and it was diagnosed as EDS-76 by serologic and virological investigations. Egg production fell suddenly when the hens were 30 to 55 weeks of age, and the depression lasted 3 to 7 weeks. Production fell 6 to 25%. Depressed egg production was accompanied by the laying of shell-less, soft-shelled, and thin shelled eggs associated with loss of egg-shell pigment. Eleven isolates of hemagglutinating adenovirus were isolated from cloacal swabs (10 isolates) and a uterus (1 isolate) of hens in one farm. One isolate, cloned and named JPA-1, had the same antigenicity in serologic tests and the same biological and physicochemical properties as the BC14 strain of EDS-76 virus. PMID- 6274292 TI - Pathogenicity and distribution of egg-drop syndrome-1976 virus (JPA-1) in inoculated laying hens. AB - Rhode Island Red laying hens that lacked hemagglutination-inhibition antibody were inoculated orally with the JPA-1 strain of adenovirus isolated from a flock affected with egg-drop syndrome-1976 in Japan and observed for 80 days. Inoculated hens laid abnormal eggs, such as shell-less, soft-shelled, and cracked eggs and those with loss of pigmentation, from 8 days postinoculation (PI). Fifteen out of 16 hens laid abnormal eggs. Egg-production rte fell from 94% to 50% between 13 and 16 days PI. When the abnormal eggs were excluded, egg production was 17%; then it recoverd slowly, reaching 67% by the end of the experiment. The virus was recovered from various organs of inoculated hens from 3 to 7 days PI. Fluorescent antigens of the virus were observed in the epithelial cells and desquamated cells from the epithelium of the uterus and isthmus 10 and 14 days PI. PMID- 6274294 TI - Serologic and cross-protection studies with several infectious bronchitis virus isolates from Delmarva-reared broiler chickens. AB - Five isolates, from broiler-type chickens, capable of causing acute respiratory disease were identified as infectious bronchitis virus (IBV). The agar-gel precipitin (AGP) test was effective in identifying the isolates as IBV. Virus neutralization (VN) and cross-challenge tests indicated two of the isolates to be Massachusetts types and one to be a JMK type. The remaining isolates were different from strains normally encountered in chickens reared on the Delmarva Peninsula. One isolate, although serologically similar to the Connecticut strain, was able to infect Connecticut-virus-immune chickens. Another isolate was similar to Arkansas 99, a strain not previously reported in Delmarva. PMID- 6274293 TI - Cross-immunity in chickens using seven isolates of avian infectious bronchitis virus. AB - Groups of 75 chickens were each infected with one of 7 isolates of infectious bronchitis virus (Mass 41, Holland 52, SE 17, Ark 99, Clark 333, JMK, and Florida). Following recovery, they were challenged along with susceptible controls to the homologous and 6 heterologous isolates. Cross-immunity was determined by virus recovery 4 or 5 days post-challenge. Challenge with the homologous isolate resulted in 90-100% protection. Challenge with heterologous isolates gave variable results and an overall average resistance of 38%. The SE 17-recovered chickens had a 50% protection, whereas the Holland-52-recovered chickens ahd a 13.3% protection. PMID- 6274295 TI - Adenovirus isolation and serology from wild bobwhite quail (Colinus virginianus). PMID- 6274296 TI - Plasma cell changes in the gland of Harder following infectious bursal disease virus infection of the chicken. AB - Tissue changes in the gland of Harder (GH) were studied following infection of day-old chicks with infectious bursal disease virus (IBDV). Broiler chicks that lacked maternal antibody to IBDV were inoculated with IBDV at 1 day of age and were maintained segregated from uninoculated controls. Plasma cell (PC) content of the GH was lowered among infected chickens from 1 to 7 weeks postinoculation. Since the GH in uninoculated control birds was not densely populated with PCs until 3 weeks of age, the PC depletion found in young chickens infected with IBDV became more clear with age. After the chickens were 3 weeks old, fewer (P less than .01) plasma cells were observed in the GH of IBDV-infected chicks than in uninoculated controls. Lymphoid follicles and heterophil populations in the GH did not appear to be affected by IBDV infection, nor could necrotic or degenerative changes be found in the acini or excretory ducts. The observation of fewer PCs suggests that a major lymphoid tissue of the upper respiratory tract is comprised by IBDV infection of day-old chicks.U PMID- 6274297 TI - Detection and quantification of antibodies to infectious bronchitis virus by enzyme-linked immunosorbent assay. AB - An enzyme-linked immunosorbent assay (ELISA) for detecting and quantifying antibodies to infectious bronchitis virus (IBV) is described. Purified antigen, prepared on sucrose density gradients, was required to decrease the nonspecific background, and saline was found to be superior to bicarbonate buffer for coating the cuvettes with antigen. The sensitivity of the test in measuring antiserum titers could be altered greatly and linearly by adjusting the protein content of the antigen. The ELISA was able to detect an antibody response to IBV infection earlier than the virus-neutralization (VN) test. Antibody titers obtained by ELISA were considerably higher than those obtained by VN. Serotypes of IBV could not be differentiated with ELISA because of extensive antiserum cross-reactivity. The utility of ELISA in studies on IBV is discussed. PMID- 6274298 TI - Cooperative control of infectious laryngotracheitis. PMID- 6274299 TI - Effectiveness of maternal antibody against challenge with infectious bronchitis viruses. PMID- 6274300 TI - Laryngotracheitis in show chickens. PMID- 6274301 TI - [Concentration of the mucosal disease (MD/VD) virus from infected cattle intestines for the agar gel precipitation (AGP) test]. PMID- 6274302 TI - [Occurrence of comedocarcinoma in dogs]. PMID- 6274303 TI - [Virus etiology of an upper respiratory organ disease in chickens]. PMID- 6274304 TI - [Suitability of ferrets for the IBR-IPV vaccine efficacy evaluation]. PMID- 6274305 TI - Purification of sphingomyelinase to apparent homogeneity by using hydrophobic chromatography. AB - Placental sphingomyelinase has been purified to apparent homogeneity by a procedure that makes extensive use of hydrophobic interaction chromatography on sphingosylphosphocholine-CH-, octyl-, hexyl- and Blue-Sepharoses. Enzyme purification is about 10000- 14000-fold over starting extract with excellent yield (usually greater than 28%). Purification of bis-4-methylumbelliferyl phosphate phosphodiesterase activity generally paralleled that of sphingomyelinase during the final stages of the procedure. The enzyme also hydrolysed bis-p-nitrophenyl phosphate, but at a lower rate compared with bis-4 methylumbelliferyl phosphate. A single major protein was observed under non denaturing conditions. Sphingomyelinase, denatured by reduction and alkylation, is composed of a major polypeptide chain with an apparent molecular weight of 89 100 on sodium dodecyl sulphate/polyacrylamide-gel electrophoresis. Two minor lower-molecular-weight components were consistently obtained at 47 500 and 30 700. These results were also obtained after maleoylation of the reduced and alkylated sample. The enzyme contains a blocked-N-terminal amino acid. An extensive search for contaminating enzymes revealed the presence of minor amounts of acid phosphatase, which were removed from the final enzyme sample. The highly purified enzyme is stable for several weeks when stored with Triton X-100 at 4 degrees C. The pure enzyme aggregates under denaturing and electrophoretic conditions and special care must be taken to ensure that hydrophobic bonding of the protein is decreased as much as possible. The reproducibility and large scale of this procedure should facilitate further study on the structure and kinetic properties of the enzyme. PMID- 6274306 TI - Apurinic endonuclease from Saccharomyces cerevisiae. AB - An endonuclease cleaving depurinated and alkylated double-stranded DNA has been purified 500-fold from Saccharomyces cerevisiae, strain MB 1052. The enzyme has an Mr of 31 000 +/- 2000, a sedimentation value of 3.2S and a diffusion coefficient of 9.5 X 10-7 cm2/s. The enzyme was active only at apurinic/apyridiminic sites, regardless of whether they were produced by heating the DNA at acidic pH or by alkylation with the ultimate carcinogen methyl methanesulphonate. Native DNA was not acted upon. U.v.-irradiated DNA and DNA treated with the ultimate carcinogen N-acetoxy-2-acetylaminofluorene were cleaved to an extent related to the extent of apurinic/apyridiminic sites. Enzymic activity was not dependent upon Mg2+, but was stimulated approx. 3-fold by 4mM Mg2+. The enzyme did not bind to DEAE-cellulose or CM-cellulose at KCl concentrations greater than 160 mM. The endonuclease was obtained free of exonuclease and 3-methyladenine-DNA glycosylase activity in five chromatographic steps. PMID- 6274307 TI - Purification and properties of adenylyl sulphate:ammonia adenylyltransferase from Chlorella catalysing the formation of adenosine 5' -phosphoramidate from adenosine 5' -phosphosulphate and ammonia. AB - Extracts of Chlorella pyrenoidosa, Euglena gracilis var. bacillaris, spinach, barley, Dictyostelium discoideum and Escherichia coli form an unknown compound enzymically from adenosine 5'-phosphosulphate in the presence of ammonia. This unknown compound shares the following properties with adenosine 5' phosphoramidate: molar proportions of constituent parts (1 adenine:1 ribose:1 phosphate:1 ammonia released at low pH), co-electrophoresis in all buffers tested including borate, formation of AMP at low pH through release of ammonia, mass and i.r. spectra and conversion into 5'-AMP by phosphodiesterase. This unknown compound therefore appears to be identical with adenosine 5'-phosphoramidate. The enzyme that catalyses the formation of adenosine 5'-phosphoramidate from ammonia and adenosine 5'-phosphosulphate was purified 1800-fold (to homogeneity) from Chlorella by using (NH(4))(2)SO(4) precipitation and DEAE-cellulose, Sephadex and Reactive Blue 2-agarose chromatography. The purified enzyme shows one band of protein, coincident with activity, at a position corresponding to 60000-65000 molecular weight, on polyacrylamide-gel electrophoresis, and yields three subunits on sodium dodecyl sulphate/polyacrylamide-gel electrophoresis of 26000, 21000 and 17000 molecular weight, consistent with a molecular weight of 64000 for the native enzyme. Isoelectrofocusing yields one band of pI4.2. The pH optimum of the enzyme-catalysed reaction is 8.8. ATP, ADP or adenosine 3'-phosphate 5' phosphosulphate will not replace adenosine 5'-phosphosulphate, and the apparent K(m) for the last-mentioned compound is 0.82mm. The apparent K(m) for ammonia (assuming NH(3) to be the active species) is about 10mm. A large variety of primary, secondary and tertiary amines or amides will not replace ammonia. One mol.prop. of adenosine 5'-phosphosulphate reacts with 1 mol.prop. of ammonia to yield 1 mol.prop. each of adenosine 5'-phosphoramidate and sulphate; no AMP is found. The highly purified enzyme does not catalyse any of the known reactions of adenosine 5'-phosphosulphate, including those catalysed by ATP sulphurylase, adenosine 5'-phosphosulphate kinase, adenosine 5'-phosphosulphate sulphotransferase or ADP sulphurylase. Adenosine 5'-phosphoramidate is found in old samples of the ammonium salt of adenosine 5'-phosphosulphate and can be formed non-enzymically if adenosine 5'-phosphosulphate and ammonia are boiled. In the non-enzymic reaction both adenosine 5'-phosphoramidate and AMP are formed. Thus the enzyme forms adenosine 5'-phosphoramidate by selectively speeding up an already favoured reaction. PMID- 6274308 TI - Purification and properties of the insulin-stimulated cyclic AMP phosphodiesterase from rat liver plasma membranes. AB - The peripheral high-affinity cyclic AMP phosphodiesterase from rat liver plasma membranes was purified to apparent homogeneity. The procedure used involved the initial purification of liver plasma membranes and the solubilization of the enzyme by using a high-ionic-strength medium. This was followed by chromatography of the enzyme on DEAE-cellulose, Affi-Gel Blue, a novel affinity column and Sephadex G-100. A 9500-fold purification of the enzyme with a 24% yield was achieved by this procedure. The purified enzyme was apparently monomeric (Mr 52000) as it exhibited identical molecular weights on analysis by gel filtration, sedimentation and sodium dodecyl sulphate/polyacrylamide-gel electrophoresis. It is suggested that the non-Michaelis kinetics exhibited by the enzyme are due to it obeying a mnemonical mechanism, where it displays Km 0.7 micrometer, Vmax. 9.1 units/mg of protein and Hill coefficient (h) 0.62. Cyclic GMP acts as a poor substrate for the enzyme, with Km 120 micrometer and Vmax. 0.4 unit/mg of protein, and also as an inhibitor of the enzyme, with I50 (concentration giving 50% inhibition) 150 micrometer when assayed at 0.4 micrometer-cyclic AMP. Inhibition by 5'-AMP is unlikely to be of physiological importance, as it is only a weak inhibitor of the enzyme (I50 47 mM assayed at 0.4 micrometer-cyclic AMP). PMID- 6274309 TI - Characterization of the phosphorylated form of the insulin-stimulated cyclic AMP phosphodiesterase from rat liver plasma membranes. AB - Incubation of intact purified rat liver plasma membranes with insulin, cyclic AMP and ATP led to the activation of the peripheral "low-Km" cyclic AMP phosphodiesterase. When (gamma-32P]ATP was included in the incubation mixture, after purification of this enzyme to homogeneity it was found to contain 1 mol of alkali-labile 32P/mol of enzyme. Treatment of the homogeneous phosphorylated enzyme with alkaline phosphatase released all of the 32P from the protein while restoring its activity to the native state. The reversibility of the activation that is achieved by the phosphorylation of this enzyme could also be demonstrated with a high-speed supernatant from rat liver. This restored the activity of the activated membrane-bound enzyme to its native state. The Ka for the cyclic AMP dependence of this process (1.6 micrometer) was unaffected by a range of ATP concentrations (1-10 mM) and by a range of membrane protein concentrations (0.2-2 mg/ml). Adenylyl imidodiphosphate could not substitute for ATP, and concanavalin A could not substitute for insulin, as essential ligands in the activation process. The purified activated enzyme exhibited Km 0.6 microM, Vmax 10.9 units/mg of protein and Hill coefficient (h) 0.47. The Vmax. for this activated enzyme was much higher than that of the native enzyme, yet h was much lower. PMID- 6274310 TI - Immunological characterization of lysyl hydroxylase, an enzyme of collagen synthesis. AB - Antibodies to pure lysyl hydroxylase from whole chick embryos were prepared in rabbits and used for immunological characterization of this enzyme of collagen biosynthesis. In double immunodiffusion a single precipitation line was seen between the antiserum and crude or pure chick-embryo lysyl hydroxylase. The antiserum effectively inhibited chick-embryo lysyl hydroxylase activity, whether measured with the biologically prepared protocollagen substrate or a synthetic peptide consisting of only 12 amino acids. This suggests that the antigenic determinant was located near the active site of the enzyme molecule. Essentially identical amounts of the antiserum were required for 40% inhibition of the same amount of lysyl hydroxylase activity units from different chick-embryo tissues synthesizing various genetically distinct collagen types. In double immunodiffusion a single precipitation line of complete identity was found between the antiserum and the purified enzyme from whole chick embryos and the crude enzymes from chick-embryo tendon, cartilage and kidneys. These results do not support the hypothesis that lysyl hydroxylase has collagen-type-specific or tissue-specific isoenzymes with markedly different specific activities or immunological properties. The antibodies to chick-embryo lysyl hydroxylase showed a considerable degree of species specificity when examined either by activity inhibition assay or by double immuno-diffusion. Nevertheless, a distinct, although weak, cross-reactivity was found between the chick-embryo enzyme and those from all mammalian tissues tested. The antiserum showed no cross-reactivity against prolyl 3-hydroxylase, hydroxylysyl galactosyl-transferase or galactosylhydroxylysyl glucosyltransferase in activity-inhibition assays, whereas a distinct cross-reactivity was found against prolyl 4-hydroxylase. Furthermore, antiserum to pure prolyl 4-hydroxylase inhibited lysyl hydroxylase activity. These findings suggest that there are structural similarities between these two enzymes, possibly close to or at their active sites. PMID- 6274311 TI - Interactions of tervalent lanthanide ions with bacterial collagenase (clostridiopeptidase A). AB - Tervalent cations of the lanthanide (rare-earth) elements reversibly inhibit bacterial collagenase (clostridiopeptidase A; EC 3.4.24.3). Sm(3+), whose ionic radius is closest to that of Ca(2+), is the most effective inhibitor, completely suppressing clostridiopeptidase activity at a concentration of 100mum in the presence of 5mm-Ca(2+). Er(3+) and Lu(3+), which both have ionic radii smaller than either Ca(2+) or Sm(3+), inhibit less efficiently, and La(3+), which is slightly larger than Ca(2+) or Sm(3+), inhibits only weakly. These findings indicate a closely fitting, stereospecific, Ca(2+)-binding pocket in clostridiopeptidase, which excludes ions that are only slightly larger than Ca(2+) [ionic radius 0.099nm (0.99 A)]. By contrast, trypsin, an enzyme whose activity does not depend on Ca(2+), requires lanthanide concentrations 50-100 fold greater for inhibition. Furthermore, the relative efficiency of inhibition of trypsin by lanthanides increases as the lanthanide ions become smaller and the charge/volume ratio increases. At a concentration of 50mum, Sm(3+) lowers the apparent K(m) for the hydrolysis of Pz-peptide by clostridiopeptidase from 5.4mm to 0.37mm and the apparent V(max.) from 0.29 Wunsch-Heidrich unit to 0.018 unit. Thus Sm(3+) enhances the affinity of this enzyme for its substrate; inhibition of hydrolysis of Pz-peptide may result from the excessive stability of the enzyme Sm(3+)-substrate complex. Inhibition by Sm(3+) is competitive with regard to Ca(2+). The apparent dissociation constant, K(d), of Ca(2+) is 0.27mm, where the K(i) for Sm(3+) is 12mum. Clostridiopeptidase is more thermolabile in the absence of Ca(2+). With Sm(3+), thermoinactivation of the enzyme at 53 degrees C or 60 degrees C is initially accelerated, but then becomes retarded as heating continues. Lanthanide ions bind to gelatin and collagen. In so doing, they appear to protect these substrates from lysis by clostridiopeptidase through mechanisms additional to supplanting Ca(2+) at its binding site on the enzyme. Collagen and gelatin sequester sufficient lanthanide ions to gain partial protection from clostridiopeptidase in the absence of an extraneous source of these inhibitors. PMID- 6274312 TI - Kinetic and e.p.r. studies on the inhibition of xanthine oxidase by alloxanthine (1 H-pyrazolo [3, 4-d] pyrimidine-4,6-diol). AB - The inhibition by alloxanthine of oxidation of xanthine by xanthine oxidase is characterized by a prolonged transient phase. Kinetic data accord with a mechanism that involves rapid formation of a reduced enzyme-alloxanthine complex that subsequently undergoes a relatively slow-reversible reaction. In this scheme the slowly formed complex cannot be fully reoxidized by oxygen. From the Ki value for the dissociation of alloxanthine from the rapidly formed complex (1.15 microM) and values of 0.37 min-1 and 0.011 min-1 for the forward and reverse rate constants of the slow reaction, an overall inhibition constant for alloxanthine of 35 nM was calculated. A molybdenum (V) e.p.r. signal from the slowly formed reduced enzyme-alloxanthine complex is described. The rate of appearance of this new signal is consistent with this assignment. The signal (the "Alloxanthine signal") was simulated with g1 2,0269, g2 1,9593, g3 11.9444 and shows indications of hyperfine coupling to nitrogen. Similarities between it and the Very Rapid signal are discussed. Close structural analogies between the catalytic intermediate represented by the Very Rapid signal and the inhibitor complex represented by the Alloxanthine signal are suggested. PMID- 6274314 TI - Nuclear association states of rat uterine oestrogen receptors as probed by nuclease digestion. AB - The solubilization of oestrogen receptors from uterine nuclei by micrococcal nuclease and deoxyribonuclease I was examined after the injection of oestradiol or Nafoxidine into castrated female rats. At 1h after an injection of oestradiol, 30% (0.18pmol/mg of DNA) of the nuclear oestrogen receptors was solubilized by 5 min of mild digestion with either nuclease. No further receptor release occurred, although DNA hydrolysis continued throughout a 20min interval. The limitation in receptor solubilization was not due to an artifact of digestion conditions or insufficient nuclease concentrations. Similar patterns of receptor solubilization and DNA hydrolysis were obtained with both nucleases whether the animals had been injected with oestradiol 1h before death or if the uteri from uninjected animals were incubated with [(3)H]oestradiol for 1h in vitro. When uterine nuclei were digested with these enzymes 12h after the animal was injected with oestradiol there was little change in the quantity of nuclease-sensitive sites (0.11pmol/mg of DNA); however, the quantity of nuclease-resistant sites decreased 10-fold. These values correspond quantitatively to the changes in salt-resistant and salt extractable sites observed over a 12h interval after oestradiol treatment. Nuclease digestion of uterine nuclei obtained 16h after Nafoxidine treatment gave a pattern qualitatively and quantitatively similar to that observed 1h after oestradiol treatment, a result consistent with the agonist/antagonist action of this compound. An analysis by sucrose-density-gradient centrifugation of the time course of nuclease-dependent receptor solubilization indicated that the solubilized receptors were not associated with discrete nucleosomal fragments. We believe that these data indicate that only a portion of the receptors translocated to the nucleus become associated with chromatin, and this association may occur on regions of chromatin that are preferentially susceptible to nucleolytic cleavage. PMID- 6274313 TI - The separate roles of glucose and insulin in the induction of glucokinase in hepatocytes isolated from neonatal rats. AB - 1. The specificity of the effect of glucose on the induction of glucokinase activity that occurs when hepatocytes freshly isolated from 13-day-old rats are incubated in Medium 199 together with insulin [Wakelam & Walker (1980) FEBS Lett. 111, 115-119] was examined. A pattern that is different from other known effects of glucose is found, and metabolism of this compound is not necessarily to account for this particular effect. 2. The effects of a raised glucose concentration and of insulin on the induction can be separated. The hexose initiates the process in the absence of insulin in a manner that is sensitive to actinomycin D but not to cycloheximide. The subsequent effect of insulin is dependent on the prior effect of glucose or other positive analogue, does not require the presence of glucose and is inhibited by cycloheximide but not by actinomycin D. 3. Induction of glucokinase in vitro in hepatocytes from neonatal animals is inhibited by adrenaline, glucagon and dibutyryl cyclic AMP, but not by vasopressin or angiotensin II. The inhibition by cyclic AMP is on the stage requiring insulin and is comparatively specific, because total protein synthesis is not apparently diminished. 4. The implications of these results are discussed with reference to possible mechanisms of induction and to the situation in vivo. PMID- 6274315 TI - The use of proteinases to determine the topological location of cytochrome P-450 in vesicles derived from smooth endoplasmic reticulum of rat liver. AB - 1. The phospholipid bilayer of intact vesicles from smooth endoplasmic reticulum is impermeable to macromolecules. Specific and non-specific proteinases were used to investigate the site of membrane proteins in the transverse plane of the bilayer. 2. When two proteinases were used in conjunction, denaturing effects additional to proteolysis were observed on cytochrome P-450 content and glucose 6 phosphatase activity which did not depend on the integrity of the phospholipid bilayer. 3. When lipid peroxidation was inhibited, these effects were not observed. PMID- 6274316 TI - Action of low-density lipoprotein and compactin, a competitive inhibitor of 3 hydroxy-3-methylglutaryl-CoA reductase, on the synthesis of dolichol-linked oligosaccharides and low-density-lipoprotein receptor in human skin fibroblasts. AB - To explore whether there is an inter-relationship between the rate of low-density (LD) lipoprotein binding to its receptor and the formation of dolichol-linked oligosaccharides, experiments were performed with human fibroblasts where the synthesis of lipoprotein receptor and dolichyl saccharides was under control of LD lipoprotein and compactin. Pretreatment of the cells with nonlabelled LD lipoprotein resulted in a suppression of both the binding of 125I-labelled LD lipoprotein to the receptor and the synthesis of dolichyl saccharides from [14C]acetate and [3H]mannose, but not from [3H]mevalonolactone. Compactin, in contrast, inhibited only the formation of dolichol-linked oligosaccharides. Mevalonolactone (1 microM) abolished the inhibitory effect of LD lipoprotein on dolichyl saccharide formation, but was not able to restore the receptor-binding capacity, thus suggesting that the synthesis of lipoprotein receptor is not coupled to the formation of dolichyl saccharides. PMID- 6274317 TI - An alternative pathway for the biosynthesis of isoprenoid compounds in bacteria. AB - The pattern of incorporation of radioactivity from [1-14C]acetate and [2 14C]acetate into the polyprenyl side-chain of ubiquinones in bacteria (Azotobacter vinelandii, Pseudomonas sesami, Escherichia coli and Rhodopseudomonas capsulata) was studied. For this purpose, a new degradation method involving a modified Barbier-Wieland reaction of laevulinic acid was developed, and used along with the iodoform reaction. Both C-1 and C-2 of acetate were incorporated exclusively into C-2 of laevulinic acid suggesting that the well-known pathway through acetoacetyl-CoA ('acetoacetate pathway') was not operative in these bacteria. An alternative pathway ('acetolactate pathway'), starting with pyruvate and acetaldehyde as the distal precursors, and utilizing the reactions of leucine and valine metabolism, was postulated. It was also postulated that C-1 of acetate is incorporated not directly, but after oxidation to CO2. The pattern of incorporation of radioactivity from [U-14C]valine, [U 14C]alanine and NaH14CO3 into the side-chain of ubiquinone of R. capsulata was in agreement with the operation of the 'acetolactate pathway'. PMID- 6274318 TI - Regulation of collagen post-translational modification in transformed human and chick-embryo cells. AB - Changes in the regulation of collagen post-translational modification in transformed cells were studied in three established human sarcoma cell lines and in chick-embryo fibroblasts freshly transformed by Rous sarcoma virus. The collagens synthesized by all but one of these and by all the control human and chick-embryo cell lines were almost exclusively of types I and/or III. The relative rate of collagen synthesis and the amounts of prolyl hydroxylase activity and immunoreactive protein were markedly low in all the transformed human cell lines. The other enzymes studied, lysyl hydroxylase, hydroxylysyl galactosyltransferase and galactosylhydroxylysyl glucosyltransferase, never showed as large a decrease in activity as did prolyl hydroxylase, suggesting a more efficient regulation of the last enzyme than of the three others. The chick embryo fibroblasts freshly transformed by Rous sarcoma virus differed from the human sarcoma cells in that prolyl hydroxylase activity was distinctly increased, whereas the decreases in immunoreactive prolyl hydroxylase protein and the three other enzyme activities were very similar to those in the simian-virus-40 transformed human fibroblasts. It seems possible that this increased prolyl hydroxylase activity is only a temporary phenomenon occurring shortly after the transformation, and may be followed by a decrease in activity later. The newly synthesized collagens of all the transformed cells that produced almost exclusively collagen types I and/or III had high extents of lysyl hydroxylation, and there was also an increase in the ratio of glycosylated to non-glycosylated hydroxylysine. The data suggest that one critical factor affecting modification is the rate of collagen synthesis, which affects the ratio of enzyme to substrate in the cell. PMID- 6274319 TI - Effect of lutropin and cycloheximide on lutropin receptors and cyclic AMP production in Leydig tumour cells in vitro. AB - A system to study lutropin-induced desensitization of tumour Leydig cells in vitro has been investigated. Tumour Leydig cells were purified on a Percoll gradient and then incubated for 30 min with lutropin (0-1000ng/ml). The cells were then washed and incubated in suspension media at 32 degrees C. 125I-labelled human choriogonadotropin binding and basal and lutropin-stimulated cyclic AMP production were determined at various times. Initially the cells showed a dose dependent decrease in human choriogonadotropin binding (1.18 and 0.13fmol/10(6) cells respectively) followed by an increase at 1 h (2.32 and 0.87fmol/10(6) cells respectively). Human choriogonadotropin binding remained elevated in the cells pre-incubated without lutropin, whereas the cells pre-incubated with lutropin showed a dose-dependent decrease over the next 10 h (2.20-0.18fmol/10(6) cells respectively). Basal production of cyclic AMP initially reflected the pre incubation conditions (1.17-21.19ng/10(6) cells per h for 0-1000ng of lutropin/ml respectively). However, by 1 h there was a marked rise in basal cyclic AMP production which returned to the initial lower values by 4 h. At all time intervals studied, lutropin-induced cyclic AMP production showed a decrease that was proportional to lutropin concentration in the pre-incubated media. The decreases in human choriogonadotropin binding produced by pre-incubations with lutropin (100ng/ml) was partially inhibited by the presence of cycloheximide in the pre-incubation media and totally prevented by the continuous presence of cycloheximide. These results demonstrate that desensitization of tumour Leydig cells occurs after exposure to lutropin in vitro. This desensitization involves both a loss of plasma membrane receptors for lutropin and lutropin-stimulated adenylate cyclase. These events can be prevented by cycloheximide and are therefore probably dependent on protein synthesis. PMID- 6274320 TI - The kinetics of the beta-galactoside-proton symport of Escherichia coli. AB - beta-Galactoside transport by Escherichia coli occurs with the concomitant uptake of a proton. The kinetics of beta-galactoside uptake at various values of external pH are interpreted in terms of a model in which both the galactoside and the proton are substrates of the transport reaction. The values of some of the kinetic constants for this two-substrate reaction were determined. The observed effects of the protonmotive force on the apparent Michaelis constant for galactoside can be explained in terms of the proton being a substrate of the transport reaction. PMID- 6274321 TI - Effects of trifluoperazine and pimozide on stimulus-secretion coupling in pancreatic B-cells. Suggestion for a role of calmodulin? AB - The possible involvement of calmodulin in insulin release was evaluated by studying the effects on intact islets of trifluoperazine and pimozide, two antipsychotic agents known to bind strongly to calmodulin in cell-free systems. Trifluoperazine (10-100mum) produced a dose- and time-dependent inhibition of the two phases of glucose-stimulated insulin release. The effect was not reversible by simple washing of the drug, but could be prevented by cytochalasin B or theophylline. Trifluoperazine also inhibited the release induced by glyceraldehyde, oxoisocaproate, tolbutamide or barium, but not that stimulated by 10mm-theophylline or 1mm-3-isobutyl-1-methylxanthine. Pimozide (0.5-10mum) also produced a dose-dependent inhibition of insulin release triggered by glucose, leucine or barium, but did not affect the release induced by methylxanthines. Glucose utilization by islet cells was not modified by trifluoperazine (25mum), which slightly increased cyclic AMP concentration in islets incubated without glucose. The drug did not prevent the increase in cyclic AMP concentration observed after 10min of glucose stimulation, but suppressed it after 60min. Basal or glucose-stimulated Ca(2+) influx (5min) was unaffected by 25mum trifluoperazine, whereas Ca(2+)net uptake (60min) was inhibited by 20%. Glucose stimulated Ca(2+) uptake was almost unaffected by pimozide. In a Ca(2+)-free medium, trifluoperazine decreased Ca(2+) efflux from the islets and did not prevent the further decrease by glucose; in the presence of Ca(2+), the drug again decreased Ca(2+) efflux and inhibited the stimulation normally produced by glucose. In the absence of glucose, trifluoperazine lowered the rate of Rb(+) efflux from the islets, decreased Rb(+) influx (10min), but did not affect Rb(+) net uptake (60min). It did not interfere with the ability of glucose to decrease Rb(+) efflux rate further and to increase Rb(+) net uptake. The results show thus that trifluoperazine does not alter the initial key events of the stimulus secretion coupling. Its inhibition of insulin release suggests a role of calmodulin at late stages of the secretory process. PMID- 6274322 TI - Cyclic AMP phosphodiesterase activity during differentiation of rabbit erythroid bone marrow cells. AB - Changes in the activity of cyclic AMP phosphodiesterase during differentiation of rabbit bone marrow erythroid cells were investigated. The cells were separated by velocity sedimentation at unit gravity into six fractions corresponding to different stages of development: proerythroblasts, basophilic cells, polychromatic cells, early orthochromatic and late orthochromatic cells and reticulocytes. Cyclic AMP phosphodiesterase was found to be very active in the most immature cells, the proerythroblasts, which also have the highest content of cyclic AMP. After differentiation into basophilic erythroblasts, a 4-fold decrease in cyclic AMP phosphodiesterase activity was observed. In these cells the amount of cyclic AMP was about 80% lower than that in proerythroblasts. In polychromatic cells a further drop in phosphodiesterase activity occurred. After the final cell division the enzyme activity was very low and the levels of cyclic AMP in the early and late orthochromatic cells remained constant. Kinetic studies demonstrated a heterogeneity of erythroid cell cyclic AMP phosphodiesterase: high affinity, low-Km (5.5 X 10(-6) M) and low affinity, high-Km (0.1 X 10(-3) M) enzymes were found. The phosphodiesterase activity was dependent on the presence of Mg2+ and was activated by Ca2+ at low Mg2+ concentrations (1 mM). The changes in cyclic AMP phosphodiesterase activity during differentiation and maturation of erythroid cells suggest the possible importance of this enzyme in the physiological control of cyclic AMP concentrations in developing erythroblasts. The loss of cyclic AMP phosphodiesterase activity after cessation of cell division supports the concept of the significance of the final cell division in erythroblast differentiation. PMID- 6274323 TI - Cyclic AMP-binding and cyclic AMP-dependent protein kinase activities in the cytosol of differentiating bone marrow erythroblasts. AB - Cytosolic cyclic AMP-binding capacity and cyclic AMP-dependent protein kinase activity have been studied in relation to differentiation and maturation of rabbit bone marrow erythroblasts. Using cells fractionated by velocity sedimentation at unit gravity, it was found that both activities decreased in dividing cells when calculated in terms of cell number but remained constant per cell volume. After the final cell division, cyclic AMP-dependent protein kinase activity did not change further, whereas cyclic AMP-binding capacity declined. There were no qualitative, but only quantitative, changes in the cyclic AMP binding proteins that are present in the cytosol of developing erythroblasts. In the immature cells, the apparent KD for the interaction of binding proteins with cyclic AMP was 4 X 10(-8) M. The data suggest that changes in cyclic AMP-binding activity during differentiation of erythroid cells are due both to changes in the amount of binding proteins and in their affinity for cyclic AMP. Plasma membranes of erythroblasts were also able to bind cyclic AMP but only in dividing cells. PMID- 6274324 TI - Plant flavokinase. Affinity-chromatographic procedure for the purification of the enzyme from mung-bean (Phaseolus aureus) seeds and conformational changes on its interaction with orthophosphate. AB - Flavokinase was purified, for the first time from a plant source [mung bean (Phaseolus aureus)] by affinity chromatography in the presence of orthophosphate and by using C-8 ATP-agarose (ATP linked through the C-8 position to beaded agarose), Cibacron Blue and riboflavin--Sepharoses. An altered substrates saturation pattern was observed in the presence of K2HPO4. The conformational changes of the enzyme in the presence of K2HPO4 were monitored by fluorescence spectroscopy. These results highlight the regulatory nature of this enzyme. PMID- 6274325 TI - Dolichyl phosphate phosphatase from Tetrahymena pyriformis. AB - A soluble dolichyl phosphate phosphatase from Tetrahymena pyriformis was purified about 68-fold. The enzyme appeared to be specific for dolichyl phosphate and existed in two interrelated forms, one of mol.wt. about 500000 and the other of mol.wt. about 63000. The enzyme was strongly inhibited by 5 mM-Mn2+ and was strongly stimulated by Mg2+. Tetrahymena in the exponential growth phase contained more of this enzymic activity than cells in stationary or lag phase. The dolichyl phosphate phosphatase may be loosely bound to mitochondrial membranes. Two roles proposed for this enzyme are (1) that of releasing dolichol from its phosphorylated biosynthetic form for its use in the cell as unesterified dolichol or dolichyl ester and/or (2) that of regulation of synthesis of glycoproteins or some other glycosylated compound. PMID- 6274326 TI - Purification and properties of human chorionic gonadotropin/lutropin receptor from plasma-membrane and soluble fractions of bovine corpora lutea. AB - Plasma-membrane and soluble fractions containing human chorionic gonadotropin/lutropin receptor were prepared from bovine corpora lutea by ultracentrifugation. The plasma-membrane and soluble fractions were studied for physicochemical properties, salts and gangliosides. The receptor preparations obtained from the plasma-membrane purified individually by sucrose-density gradient centrifugation, which resulted in a partial dissociation of the hormone binding subunit from the intact functional receptor unit, which consists of both hormone-binding (regulatory) and adenylate cyclase-associated (catalytic) subunits. The fractions containing the functional receptor unit were further purified by gel filtration on Sepharose-6B and chromatography on concanavalin A Sepharose. The 'receptor' was finally purified by affinity chromatography on a column of controlled-pore glass covalently coupled to hu man chorionic gonadotropin. The purified receptor from the plasma-membrane and the soluble fractions contained binding capacities of 901000 and 87000 fmol of human chorionic gonadotropin/mg of protein. Yields of 0.02 and 0.22mg of protein were obtained from 250 g of bovine corpora lutea, which represents a 10000- and 1000 fold increase respectively in the specific binding with 125I-labelled human chorionic gonadotropin. Immunization of rabbits with a partially purified receptor fraction generated antibodies that specifically inhibited the binding of the 125I-labelled human chorionic gonadotropin to the receptor. PMID- 6274327 TI - Inhibition of rabbit tissue kininase by anti-(endo-oligopeptidase A) antibodies. AB - Highly purified rabbit brain endo-oligopeptidase A injected into goats produced, after 60 days of immunization, antisera that specifically inhibit purified rabbit brain endo-oligopeptidase A. An immunoreactive kininase having the same specificity as rabbit brain endo-oligopeptidase A for bradykinin was detected in several rabbit tissues. The highest amount of this immunoreactive kininase was found in the 25000 g supernatant fraction (S fraction) of heart, liver, skeletal muscle, ovary, brain and testis homogenates, corresponding to 89, 86, 78, 59, 56 and 53% respectively of the whole kininase activity found in the S fraction. PMID- 6274328 TI - Adenosine deaminase activity and superoxide formation during phagocytosis at different cell densities. PMID- 6274329 TI - Angiotensin-converting enzyme measured in mouse tissue by inhibition of histidyl leucine peptidase. PMID- 6274331 TI - Inhibition of epidermal growth factor binding by prostaglandin A's. PMID- 6274330 TI - Thromboxanes and prostacyclin: positive and negative modulators of tumor growth. PMID- 6274332 TI - Interaction of the inhibitor of the 3',5' cyclic AMP phosphodiesterase of Dictyostelium discoideum with the Affi-Gel blue. PMID- 6274333 TI - Occurrence of the adenylate cyclase "G protein" in membranes of Dictyostelium discoideum. PMID- 6274334 TI - Inhibition of liver glycogen synthase phosphatase by calcium: new evidence for an interaction between synthase activation and phosphorylase a. PMID- 6274335 TI - UV light-induced cross-linking of strychnine to the glycine receptor of rat spinal cord membranes. PMID- 6274336 TI - Adenocarcinoma of the human exocrine pancreas: presence of secretin and caerulein receptors. PMID- 6274337 TI - Interfacial spin trapping in model membrane systems. PMID- 6274338 TI - Effects of aphidicolin on retrovirus DNA synthesis in vivo. PMID- 6274339 TI - DNA methylation in normal and SV40-transformed human fibroblasts. PMID- 6274340 TI - Vanadate increases oxygen affinity and affects enzyme activities and membrane properties of erythrocytes. PMID- 6274341 TI - Studies on brain opiate receptor stability in vitro: inhibition of opiate receptor binding by adenosine-5'-diphosphate. PMID- 6274342 TI - Similarities in the mechanisms by which formyl-methionyl-leucyl-phenylalanine, arachidonic acid and leukotriene B4 increase calcium and sodium influxes in rabbit neutrophils. PMID- 6274343 TI - The src gene product (pp60 src) of avian sarcoma virus rapidly induces DNA synthesis and proliferation of calcium-deprived rat cells. PMID- 6274344 TI - Effect of 1,4-dihydroxy-5,8-bis (2-(2-hydroxyethylamino) ethyl amino)-9,10 anthracenedione (dihydroxyanthraquinone) on the replication of simian virus 40 chromosome. PMID- 6274345 TI - Chick bone collagenase inhibitor and latency of collagenase. PMID- 6274346 TI - Inactivation of phosphofructokinase 2 by cyclic AMP - dependent protein kinase. PMID- 6274347 TI - Poly (ADP-ribose) synthetase activity in rat testis mitochondria. PMID- 6274348 TI - Steric factors involved in the action of glycosidases and galactose oxidase. PMID- 6274349 TI - Induction of 2,3-bisphosphoglycerate synthase in Friend leukemia cells. PMID- 6274350 TI - Molecular actions of angiotensin. PMID- 6274351 TI - Imidazole derivatives as inhibitors of cytochrome P-450-dependent oxidation and activators of epoxide hydrolase in hepatic microsomes from a marine fish. PMID- 6274352 TI - Anthracenedione activation by NADPH-cytochrome P-450 reductase; comparison with anthracyclines. PMID- 6274353 TI - The molecular basis of neurotransmission at the muscarinic receptor. PMID- 6274354 TI - Nitroreductase-induced binding of nitroaromatic radiosensitizers to unsaturated lipids. Nitroxyl adducts. PMID- 6274355 TI - Inhibition of benzodiazepine receptor binding by several tryptophan ad indole derivatives. PMID- 6274356 TI - Inhibition of dog brain synaptosomal Na+ -K+ ATPase and K+-stimulated phosphatase activities by long chain n-alkyl-amine and -piperidine, and N'-alkylnicotinamide derivatives. PMID- 6274357 TI - Changes in brain polyphosphoinositide metabolism induced by cationic amphiphilic drugs in vitro. PMID- 6274358 TI - Effect of heparin on the subcellular distribution of human placental 7 ethoxycoumarin O-deethylase activity. PMID- 6274360 TI - Induction of lupus inclusions by BUdR is mediated by a human lymphoid cell factor. PMID- 6274359 TI - Release of collagenase, neutral protease, and prostaglandins from cultured mammalian synovial cells by hydroxyapatite and calcium pyrophosphate dihydrate crystals. AB - Hydroxyapatite (HA) and calcium pyrophosphate dihydrate (CPPD) crystals were phagocytosed when added to cultured human rheumatoid or normal canine synovial cells. Collagenase and neutral protease secretion into the culture medium was increased 5- to 8-fold over control values in the presence of HA and increased 3 fold in the presence of CPPD crystals. HA but not CPPD crystals induced a 300 fold increase in human rheumatoid synovial cell culture fluid prostaglandin (PG) E2 levels and an 8-fold increase in PGF alpha levels. This mechanism may be important in the pathogenesis of the destructive arthropathies associated with HA and CPPD crystals. PMID- 6274361 TI - [Moxalactam and cefotaxime--two new beta-lactam antibiotics compared with three well-known cephalosporins]. AB - This study compares the minimum inhibitory concentrations of moxalactam and cefotaxime, two beta-lactam-antibiotics, with those of cefoxitin, cefuroxime, and cefazolin for 523 gram-positive and -negative strains. Moxalactam and cefotaxime inhibit nearly all clinically important Enterobacteriaceae at concentrations less than 0.05 microgram/ml. Cefoxitin, cefuroxime, and cefazolin suppress bacterial growth at about 5--10 times higher concentrations. All strains of Pseudomonas are normally resistant to those three established cephalosporins, but about 60% are susceptible to 32 micrograms/ml of cefotaxime and moxalactam, a concentration which falls below achievable serum levels however does not exhibit any therapeutical effect. Against Staphylococcus aureus cefazolin is the most effective of all antibiotics tested (MIC 0.125--1 microgram/ml), cefotaxime appears to be somewhat less active (MIC 2--4 micrograms/ml), and a minimum of 8 micrograms/ml of moxalactam is needed to prevent bacterial growth. Streptococcus faecalis seems to be resistant to 128 micrograms/ml of moxalactam whereas the effect of cefotaxime is quite different. After transfers of Enterobacteriaceae on media containing cefotaxime or moxalactam a striking decrease of susceptibility sometimes occurs. PMID- 6274362 TI - Pharmacokinetics of pipecurium bromide, a new non-depolarizing neuromuscular blocking agent, in humans. AB - The pharmacokinetics of 2 beta, 16 beta-bis-(4'-dimethyl-1'-piperazino)-3 alpha, 17 beta-diacetoxy-5 alpha-androstane dibromide (pipecurium bromide, Arduan), a new non-depolarizing neuromuscular blocking agent, was studied in surgical patients. Plasma concentration-time curves of pipecurium bromide were evaluated by fitting the data to a bi-exponential equation. The average half-lives were T 1/2 alpha = 4.1 +/- 1.4 (n = 8) and T 1/2 beta = 44 +/- 7 (n = 8) min. The pharmacokinetic parameters of pipecurium bromide were found as follows: apparent distribution volume V beta = 261 +/- 28 (n = 8) ml/kg, plasma clearance Cl = 320 +/- 55 (n = 8) ml/min. The microconstants of two-compartment open models were also calculated. PMID- 6274363 TI - Influence of plafibride, an antiplatelet and hypolipemic agent, on prostacyclin and thromboxane synthesis, 3',5'-cyclic AMP phosphodiesterase activity and serum clearance of a lipid emulsion. AB - The activity of N-2-(p-chlorophenoxy)-isobutyryl-N'-morpholinomethylurea (plafibride, ITA 104) on arachidonic acid metabolism, the 3',5'-cyclic AMP phosphodiesterase and the serum clearance of a lipid emulsion is reported in order to clarify its mechanism of action on platelet aggregation and lipid metabolism. Plafibride did not act on the arachidonic acid metabolism as far as platelet aggregation was concerned, since it did not modify the generation of prostaglandin endoperoxides nor prostacyclin. Neither did it act on the generation of thromboxane A2. Plafibride inhibited the activity of 3',5'-cyclic AMP-phosphodiesterase, which is one of the principal mechanisms of inhibition of platelet aggregation. The serum c clearance of a commercial lipid emfy the generation of prostaglandin endoperoxides nor prostacyclin. Neither did it act on the generation of thromboxane A2. Plafibride inhibited the activity of 3',5' cyclic AMP-phosphodiesterase, which is one of the principal mechanisms of inhibition of platelet aggregation. The serum clearance of a commercial lipid emulsion was increased by plafibride which also possessed a strong hypotriglyceride activity. The correlation between platelet antiaggregant and hypolipemic activity of plafibride is discussed in this paper. PMID- 6274364 TI - [Bacteriological in vitro studies with a new cephalosporin: cefotiam (author's transl)]. AB - MIC determinations with the dilution method using DST-agar show that 7 beta-[2 amino-thiazol-4-yl)acet-amido-3-less than ([1-(2-dimethylaminoethyl)-1H-tetrazol 5-yl]thio)methyl greater than-ceph-3-em-4-carboxylic acid (cefotiam) is approximately ten times more active than cefoxitin or cefuroxime against non selected bacterial isolates. Cefotiam is also significantly more active than cefoxitin or cefoperazone against mezlocillin- and piperacillin-resistant cultures. The determination of susceptibility to cefotiam gives reliable results with 30 microgram discs. Inhibition zones of up to 12 mm (corresponding to MIC superior or equal to 64 microgram/ml) should be considered as resistance and zones larger or equal to 18 mm (corresponding to MIC inferior or equal to 8 microgram/ml) as susceptibility. PMID- 6274365 TI - [A human pancreatic glucagonoma, ultrastructural, immunocytochemical and radioimmunological investigations (author's transl)]. AB - The authors report a case of glucagonoma in a 52 years old man presenting a migratory necrolytic erythema. By conjugated means of arteriography and splenoportography with plasma glucagon assays the tumour was localized in the tail of the pancreas. Surgical excision was easy but hepatic metastases revealed the malignant nature of the tumor. This glucagonoma has been investigated by several approaches including electron microscopy, immunocytochemistry and radioimmunological techniques. The tumor contained scattered glucagon and pancreatic polypeptide immunoreactive cells; insuline, glucagon, somatostatin, pancreatic polypeptide, gastrin and VIP antisera gave negative results. Ultrastructurally, these cells showed atypical secretory granules different from A granules of the normal glucagon cell. Radio immunological determinations carried out after gel permeation chromatography of plasma revealed high molecular weight (4 000, 9 000, 14 000) immunoreactive glucagon peptides. They have been thought to be proglucagon forms which did not react with specific antiglucagon sera used in cytological studies. Reported data are consistent with the classification of this tumor in the category of glucagonoma with the "glucagonoma syndrome". PMID- 6274367 TI - [Dependence of blood platelet aggregation capacity on agents affecting the intrathrombocyte Ca2+ content]. AB - The effect of agents changing absorption and redistribution of Ca2+ in platelets on platelet aggregation was studied. It was shown that inhibition of intercellular Ca2+ absorption by verapamil and tetracain block of intracellular Ca2+ redistribution decrease platelet aggregation. A joint action of verapamil and tetracain increase the inhibition of aggregation. EDTA decreases platelet aggregation affecting platelet regulatory mechanisms associated with the action of cAMP and probably directed at a decrease in the platelet cytoplasmic Ca2+ content. PMID- 6274366 TI - [Perspectives on immunologic damage]. AB - The most important steps followed by the science of Immunology since the end of last century to attain the knowledge on the subpopulations of T and B lymphocytes are summarized. General immunologists observed that the immune response could have adverse effects to the host. First, it was the observation of anaphylaxis and the reaction to tuberculin; next, the term hypersensitivity was coined and finally, four different mechanisms of the immunologic harm were characterized. Through the last years, a fifth hypersensitivity mechanism has been proposed where of the function of the cells having membrane receptors are damaged and against which, autoantibodies are formed. This autoimmune response may block or may stimulate the receptors. The author analyses the perspectives had by this mechanism of "cellular function altered by autoantibodies against membrane receptors". An important tissular damage does not exist like in the other hypersensitivity mechanisms, but there is rather a change in the biological activity of a tissue, due to the fact that several of its receptors become blocked or are stimulated. PMID- 6274368 TI - [Lidocaine action on the ion currents of normal and depolarized myocardial fibers in membrane potential fixation]. AB - The effect of lidocaine on the transmembrane ion currents of normal and partially depolarized atrial trabecula was studied in the frog by fixation of membrane potential with duoble sucrose bridge. Lidocaine in a concentration below 12 X 10( 4) g/ml produced no effect on the slow input current. Lidocaine decreased the potassium conduction at the membrane potential values below--45 mW, and increased at--30 mW. Kinetic parameters of the rapid input sodium current and its amplitude significantly changed under the action of lidocaine in normal cardiac tissue, and its effect increased in the depolarized tissue. It is suggested that depolarization is a factor increasing sensitivity to lidocaine of rapid sodium canals, while the capacity of lidocaine to inhibit effectively the rapid sodium current in the partially depolarized myocardium determine its antiarrhythmic activity. PMID- 6274369 TI - [Effect of cold stress on the concentrations in the hypothalamus, pituitary, and blood of beta-lipotropin and beta-endorphin in the rabbit]. AB - To evaluate the rabbit availability in beta-Lipotrophin studies we have investigated their brain, pituitary and plasmatic concentrations in basal conditions and after cold-stress. Previous silicic Acid extract and Sephadex G-75 column chromatography, the two peptides were measured through two specific and sensitive RIAs. The results reveal the presence of beta LPH and beta EP and their increase after cold-stress in hypothalamus, pituitary and peripheral plasma concentrations while not in midbrain. PMID- 6274370 TI - [Chromatography of membrane proteins in sodium dodecyl sulfate. II) Membrane proteins of some human cutaneous tumors]. PMID- 6274371 TI - [Oblongata in the spinal trigeminal nucleus of the adult rat (author's transl)]. AB - The substantia gelationsa has been noted as a region of central pain control, In 1979, we reported that an intense thiamine monophosphatase (TMPase) activity was specifically localized on the plasma membrane of the synaptic glomeruli, especially dense sinusoid axon terminals (DSA). In the present investigation, the ultracytochemical localization of TMPase activity was studied in the spinal trigeminal nucleus of the adult rat. Vibratome sections of aldehyde-fixed specimens were incubated in the incubation medium (veronal-acetate buffer, pH 5. 8, 27 mM, thiamine monophosphoric acid chloride 2mM, lead nitrate 2.7 mM, sucrose 146 mM, final pH 5.4) for 30 min at 37 degrees C, and processed for light as well as electron microscopy. At the light microscopic level, the reaction products, lead sulfide, were observed to be localized in the substantia gelatinosa of the caudal medulla oblongata in the spinal trigeminal nucleus. At the electron microscopic level, the TMPase activity was positive on the plasma membrane of DSA in the region where is particularly concerned with the relay of thermal and noxious stimuli. It has been suggested by us previously that there may be some relationship between the TMPase activity and pain, mechanism in the spinal cord, and the results that the situation seems to be hold true in the spinal trigeminal nucleus, also. PMID- 6274372 TI - Effects of glycopyrrolate and atropine combined with antacid on gastric acidity. AB - Glycopyrrolate, atropine or saline were administered i.v. with oral magnesium trisilicate 1 h before induction of anaesthesia to patients about to undergo Caesarean section. Both anticholinergics greatly reduced gastric acidity. It is suggested that glycopyrrolate should be used in combination with antacid therapy before obstetric anaesthesia and puerperal tubal ligation because of its prolonged duration of action, effect on gastric secretion and failure to cross the placental barrier. PMID- 6274373 TI - Recovery from pancuronium blockade in the neonatal intensive care unit. AB - Neuromuscular blockade with pancuronium and its antagonism was evaluated in 33 critically ill infants. The evoked contraction of the adductor pollicis from indirect stimulation of the ulnar nerve was measured. The neuromuscular blockade recovered spontaneously from pancuronium in seven infants, 23 required one or more doses of atropine 0.02 ng kg-1 and neostigmine 0.06 mgkg-1. In three infants the blockade failed to reverse. Immature infants less than 32 weeks did not show any significant different in their requirement for pancuronium compared with mature infants. Age and birth weight of the infant, dose of pancuronium and duration of its administration did not affect the requirements for reversal. Train-of-four and tetanus; twitch ratios were lower (P less than 0.05) in infants less than 32 weeks of developmental age reflecting immaturity of neuromuscular transmission. PMID- 6274374 TI - Radiotherapy alone or with chemotherapy in the treatment of small-cell carcinoma of the lung: the results at 36 months. 2nd report to the Medical Research Council on the 2nd small-cell study. AB - This report compares the results at 36 months for 121 patients treated with radiotherapy alone (R) and 115 with radiotherapy followed by 3-drug chemotherapy (RC) for small-cell carcinoma of the lung of "limited" extent. The RC patients had an increased survival (P = 0.009 by log-rank test). The median survival was 25 weeks for the R patients and 43 weeks for the RC patients, but at 36 months, only 4 (3%) of the R patients and 5 (4%) of the RC patients were still alive. There was evidence of recurrence of the primary cancer in 41 (35%) of the 117 R and 35 (32%) of the 110 RC patients who died. Distant metastases were more frequent in the R series, being reported in 99 (82%) compared with 82 (71%) of the RC patients (P less than 0.05 by log-rank test). The numbers of R patients alive and considered free of metastases were 10 (8%) at 12 months, 3 (2%) at 24 months and 3 (2%) at 36 months; the corresponding figures for the RC patients being 30 (26%), 9 (8%), and 4 (3%). PMID- 6274375 TI - Breast cancer: a comparison of response to endocrine therapy and oestrogen excretion patterns including unusual metabolites. AB - The urinary excretion patterns of oestrogen metabolites, including unusual metabolites, were determined by gas chromatography and mass spectrometry for 63 women with advanced breast cancer and 39 normal postmenopausal women. The concentration of total unusual metabolites excreted was found to be an excellent discriminant between breast-cancer patients and controls (P less than 0.0001). Discrimination between responders and non-responders to endocrine therapy was attempted, using several different indices. Of these, the ratio of Classical Oestrogens to Unusual Metabolites (CE/UM) proved a fair discriminant, but the product of this ratio and the oestriol ratio (CE/UM x E3R) was much the best discriminant. This product, termed a Pattern Index, has considerable potential, not only as a discriminant for selecting therapy, but also as a rapid index of patient response to that therapy. PMID- 6274376 TI - Experimental ulcerative herpetic keratitis. IV. Preliminary observations on the efficacy of a herpes simplex subunit vaccine. AB - Systemic vaccination of rabbits with an inactivated type 1 virus subunit vaccine induced humoral and cell-mediated immune responses. Following ocular infection with type 1 herpes virus corneal ulceration and virus excretion were reduced in the vaccinated rabbits. PMID- 6274377 TI - Wheat bran increases high-density-lipoprotein cholesterol in the rat. AB - 1. Five groups of male Sprague Dawley rats were given for 6 months a diet with high protein and fat contents but a very low dietary fibre content (group B), and this diet mixed with (g/kg) 50 low-methoxyl pectin (group L), 50 high-methoxyl pectin (group H), 50 guar gum (group G) and 200 wheat bran (group WB, corresponding to 100 wheat fibre) respectively. 2. The weight increment was significantly lower in group G than in the other groups. Assuming no energy value of the dietary fibre, the weight increment (/kJ) was the same in groups B, L and H, lower in group G and higher in group WB, indicating that a proportion of the bran fibre might in fact be available as a source of energy. 3. Wheat bran increased total plasma cholesterol and high-density-lipoprotein cholesterol after 6, 12 and 26 weeks. Group G had significantly lowered plasma cholesterol after 12 weeks. Pectin on the other hand did not significantly influence total or HDL cholesterol levels. It is therefore possible that the plasma cholesterol lowering effect of pectin previously demonstrated in the rat is dependent on the presence of significant amounts of dietary cholesterol as our diets did not contain added cholesterol. Plasma triglycerides decreased with age but were similar in all groups. PMID- 6274378 TI - Effects of removal of the forestomach and caecum on the utilization of dietary urea in golden hamsters (Mesocricetus auratus) given two different diets. AB - 1. Surgical removal of the forestomach, the caecum or both the forestomach and the caecum, was performed on growing male golden hamsters (Mesocricetus auratus) given a low-or high-fibre diet. 2. In Expt 1, 18 d after surgery, the hamsters which were given a concentrate before surgery were given an experimental low fibre diet containing urea or [15N] urea (10 g/kg diet) for 9 d. In Expt 2, 5 d after surgery, the hamsters which were given a forage diet before surgery were adopted to an experimental high-fibre diet containing urea (10 g/kg) and dried Italian ryegrass (Lolium italicum) powder (400 g/kg diet) for 5 d, and 10 d after surgery, given a diet containing [15N]urea (10 g/kg diet) for 9 d. 3. In both experiments, removal of the caecum resulted in a significant lowering of both weight-gain and the digestibility of the dry matter, and removal of the forestomach resulted in a significant decrease of both accumulation of 15N in the body protein and proportion of 15N retained in the body protein as a proportion of the administered dosage. 4. The effect of removal of the caecum on the utilization of [15N] urea was not apparent in Expt 1, but the utilization of [15N] urea was significantly lowered by removal of the caecum in Expt 2. 5. These results suggest that the caecum may play a more important role in food utilization than the forestomach. However, the forestomach of the golden hamster plays a significant role in the utilization of dietary urea. It is presumed that the role of the caecum in the utilization of urea may vary under different dietary conditions. PMID- 6274379 TI - Association of the thyroid hormone receptor with rat liver chromatin. AB - We have investigated the association of the triiodothyronine (T3) nuclear receptor with rat liver chromatin by the use of selective endonuclease digestion and differential solubilization. The T3 receptor was found in a fraction of chromatin having some of the characteristics of active chromatin: (a) It is highly sensitive to DNase I and micrococcal nuclease digestion; (b) it is enriched in nonhistone proteins and depleted of histone (H-1). Micrococcal nuclease and pancreatic DNase I excised two receptor-containing fragments from chromatin, a minor (12--14 S) form and a major (5.5--6.0 S) form. The latter structure has a Stokes radius of 42 +/- 2 A and an estimated molecular weight of 95400 when a partial specific volume of 0.725 cm3/g for protein is used. It contains DNA but no histones. Similar receptor-containing fragments were excised from chromatin of other rat tissues, including brain, kidney, and heart. Both the 5.5--6 S and the 12--14 S receptor-containing chromatin structures are converted by 0.5 M KCl to the 3.5 S form (R0 35 A molecular weight 50500). Titration with micrococcal nuclease and pancreatic DNase I revealed that the 5.5--6 S form is preferentially excised by endonuclease. Neither receptor occupancy nor thyroidal status had an apparent effect on the susceptibility of chromatin to endonucleolytic digestion nor did they influence the distribution of T3 receptors in chromatin. Our results suggest that T3 receptors are not tightly associated with nucleosomes, the basic subunit of chromatin, but are associated with the DNA linking nucleosomes in structurally modified regions of chromatin in rat liver nuclei. The T3 receptor-containing fragment may well represent a higher order of structural complexity necessary for T3 action at the cellular level. PMID- 6274380 TI - Interactions of wild-type and mutant M protein of vesicular stomatitis virus with viral nucleocapsid and envelope in intact virions. Evidence from [125I]iodonaphthyl azide labeling and specific cross-linking. AB - Four different temperature-sensitive M protein mutants (tsM) of vesicular stomatitis virus (VSV) were characterized with regard to the association of the mutated M protein either with nucleocapsids or with membranes in the intact virions. Virions were labeled with the photoreactive hydrophobic probe [125I]iodonaphthyl azide (INA) to assess interactions between viral proteins and the lipid envelope. In wild type (wt) virions, the three major structural proteins--G, M, and N--were labeled in the ratio ca. 1.0:0.4:0.2. INA labeled only the membrane-associated peptide of G protein, both in the intact virion and in reconstituted G protein--viral lipid vesicles, demonstrating the specificity of INA for lipid bilayer regions. Labeling of tsM virions with INA resulted in a 2--3-fold greater incorporation into M protein than was found for wt virions, suggesting increased M--membrane associations in the mutant virions. Temperature stable revertants from tsM possessed wt labeling characteristics. Interaction of the M protein with nucleocapsids was assessed from the abundance of disulfide linked M--N complexes found after disruption of the virions by sodium dodecyl sulfate solution under nonreducing conditions. The abundance of such complexes was 30--80% less from tsM virions than from wt virions, suggesting decreased M- nucleocapsid interactions in tsM virions. Temperature-stable revertants from tsM resembled wt in the abundance of M--N complex formed. We conclude that the mutations alter M protein in such a way as simultaneously to increase its association with membrane and to decrease its affinity for nucleocapsids in the intact virion. PMID- 6274381 TI - Diffusion-driven mechanisms of protein translocation on nucleic acids. 2. The Escherichia coli repressor--operator interaction: equilibrium measurements. PMID- 6274382 TI - Liposome-mediated delivery of deoxyribonucleic acid to cells: enhanced efficiency of delivery related to lipid composition and incubation conditions. AB - Delivery of liposome-encapsulated simian virus 40 (SV40) DNA to African green monkey Related to been used as a probe to study liposome--cell interactions and to determine conditions which favor the intracellular delivery of liposome contents to cells. The efficiency of DNA delivery by various liposome preparations (monitored by infectivity assays) was found to be dependent both on the magnitude of vesicle binding to cells and on the resistance of liposomes to cell-induced leakage of contents. Acidic phospholipids were much more effective in both binding and delivery, and phosphatidylserine (PS) was the best in both aspects. The inclusion of 50 mol % cholesterol in liposomes reduces the cell induced leakage of vesicle contents (2--5-fold) and substantially enhances the delivery of DNA to cells (2--10-fold). Following incubation of cells with negatively charged liposomes containing SV40 DNA, infectivity can be enhanced greatly by brief exposure of the cells to glycerol solutions. In contrast, only slight enhancement by glycerol was observed for SV40 DNA encapsulated in neutral or positively charged liposomes. The results of competition experiments between empty phosphatidylcholine liposomes and DNA-containing PS liposomes also suggest possible differences in the interaction of neutral and negatively charged liposome preparations with cells. Morphological studies indicate that the glycerol treatment stimulates membrane ruffling and vacuolization and suggest that the enhanced uptake of liposomes occurs by an endocytosis-like process. Results obtained with metabolic inhibitors are also consistent with the interpretation that the enhancement of liposome delivery in glycerol-treated cells occurs via an energy-dependent endocytotic pathway. Pretreatment of cells with chloroquine, a drug which alters lysosomal activity, further enhanced infectivity in glycerol-treated cells (4-fold). This observation suggests the involvement of a lysosomal processing step at some point in the expression of liposome-encapsulated DNA and, more importantly, illustrates the possibility of altering cellular mechanism to engineer more efficient delivery by liposomes. Under optimal conditions determined in this study, the efficiency of liposome mediated SV40 DNA delivery was increased more than 1000-fold over that obtained by simply incubating cells with liposomes. It is also demonstrated that these conditions enhance delivery of other molecules, besides DNA, which are encapsulated in liposomes. PMID- 6274383 TI - Fluorescent probe of ribonuclease A conformation. AB - The reaction of ribonuclease (RNase) with N-[[(iodoacetyl)amino]ethyl]-5 naphthylamine-1-sulfonic acid (1,5-IAENS) yields a derivative in which one fluorescent group is covalently attached to the protein. Several arguments suggest that the chemical modification has occurred at the enzyme active site: (i) 1,5-IAENS should have the same specificity as iodoacetamide, i.e., carboxymethylate one histidine of the active site; (ii) the derivatized protein is enzymatically inactive; (iii) in the native state of the protein, the fluorescent group is (almost) completely protected from the aqueous solvent; (iv) this group has no motions other than those of the protein. The fluorescence properties of the derivatized RNase change markedly upon unfolding induced by guanidine hydrochloride (Gdn . HCl), as seen from fluorescence intensity, maximum emission wavelength, and polarization measurements. Upon Gdn . HCl-induced unfolding, the fluorescent group is transferred from a nonpolar to a highly polar environment. Dynamic fluorescence measurements show also that unfolding results in a markedly increased mobility of the fluorescent label with respect to its proteic environment. These results are compared to those of Young & Potts (1963) [Young, D. M., & Potts, J. T. (1963) J. Biol. Chem. 238, 1995--2002], who studied the fluorescence properties of a surface-labeled derivative of RNase. PMID- 6274384 TI - Hydroxylamine oxidoreductase: a 20-heme, 200 000 molecular weight cytochrome c with unusual denaturation properties which forms a 63 000 molecular weight monomer after heme removal. PMID- 6274385 TI - Covalent complex between yeast cytochrome c and beef heart cytochrome c oxidase which is active in electron transfer. PMID- 6274386 TI - Purification and properties of poly(ADP-ribose) polymerase from lamb thymus. AB - Poly(ADP-ribose) polymerase was purified 2900-fold from lamb thymus with a recovery of 5%. Addition of exogenous DNA was essential for activity of the purified enzyme, and the reaction was stimulated by the addition of either a mixture of histones or purified histone H1. The enzyme is inhibited by sulfhydryl binding agents such as phenylmethanesulfonyl fluoride or N-ethylmaleimide. It does not require magnesium or other metal ion cofactors for activity. The enzyme migrated as a single polypeptide chain with an apparent molecular weight of 135 000 when gel electrophoresis was performed in the presence of sodium dodecyl sulfate. The apparent molecular weight was 175 000 when determined by gel filtration on Sepharose CL-6B-200. The isoelectric point was pH 9.6, and the pH optimum for activity was 8.6-8.8. The apparent Km for NAD+ was 160 microM at 37 degrees C. The activity of the purified polymerase was unaffected by the presence of ADP-ribose, 3',5'-cAMP, or NaF. Nicotinamide, 5-methyl-nicotinamide, theophylline, and thymidine markedly inhibited enzyme activity. Lamb thymus DNA, originally associated with the enzyme, was more effective than commercially obtained calf thymus DNA as an enzyme activator. PMID- 6274387 TI - Purification of rabbit brain endooligopeptidases and preparation of anti-enzyme antibodies. AB - Endooligopeptidase A was purified approximately 3 000-fold and endooligopeptidase B approximately 1200-fold from the 25 000g supernatant fraction of rabbit brain homogenate. The purified enzymes were homogeneous on the basis of acrylamide gel electrophoresis under denaturing and nondenaturing conditions, isoelectric focusing, immunochemical criteria, and specific activities of the elution profile of gel filtration on Sephadex G-100. The only peptide bond cleaved by endooligopeptidase A in bradykinin, Arg1-Pro2-Pro3-Gly4-Phe5-Ser6-Pro7-Phe8-Arg9, is Phe5-Ser6, whereas endooligopeptidase B hydrolyzes the Pro7-Phe8 peptide bond of bradykinin and the Pro3-Gly4 bond of des-Phe8-Arg9-bradykinin. The specific activity of the homogeneous enzymes using bradykinin as substrate was 1087 nmol min-1 mg-1 for endooligopeptidase A and 292 nmol min-1 mg-1 for endooligopeptidase B. Gel filtration suggested molecular weights of 75 000 and 68 000 for endooligopeptidases A and B, respectively. Sodium dodecyl sulfate gel electrophoresis suggested that each endooligopeptidase consisted of a single polypeptide chain with molecular weights of 74 000 and 69 000 for the A and B enzymes, respectively. Purified endooligopeptidase A or B injected into goats produces monospecific antisera directed against each enzyme. The antibody prepared against each endooligopeptidase did not react with or inhibit the activity of the other enzyme. PMID- 6274388 TI - Characterization of the major apolipoproteins secreted by two human hepatoma cell lines. AB - Two newly described human hepatoma derived cell lines, Hep G2 and Hep 3B [Knowles, B. B., Howe, C. C., & Aden, D. P. (1980) Science (Washington, D.C.) 209, 497-499], synthesize and secrete into the culture medium most of the major plasma apoproteins (apoA-I, apoA-II, apoB, apoC-II, apoC-III, and apoE). The synthesized apoproteins were identified by direct two-dimensional gel analysis of the culture medium or by two-dimensional analysis following purification of the apoproteins by ultracentrifugation or immunoprecipitation. We found that the apoA I synthesized by both of the hepatoma cell lines consists of two isoproteins designated 2 and 3 which are more basic than the major plasma apoA-I isoproteins designated 4 and 5. The apoE synthesized by both cell lines is composed mainly of an array of isoproteins with increasingly higher molecular weights and lower isoelectric points as compared to those of the major apoE isoproteins found in plasma. These precursors of apoE are converted to the major apoE isoproteins upon treatment with Clostridium perfringens neuraminidase and represent sialo apoE isoproteins. ApoA-II, apoC-II, apoC-III-1, and apoC-III-2 correspond to the protein forms present in plasma. The human hepatoma cell lines (Hep G2 and Hep 3B) provide a unique model for studies of the regulation of human apoprotein and lipoprotein synthesis and catabolism. PMID- 6274389 TI - Effect of sterol structure on acyl chain ordering in phosphatidylcholine vesicles: a deuterium nuclear magnetic resonance and electron spin resonance study. PMID- 6274390 TI - Spin-label detection of hemoglobin-membrane interaction at physiological pH. AB - The interaction between hemoglobin and the cytoplasmic surface of human erythrocyte membranes at physiological pH was studied by monitoring the electron paramagnetic resonance (EPR) signal of spin-labeled membrane ghosts in hemoglobin solutions of various concentrations. The EPR spectra indicate the existence of a significant hemoglobin-membrane interaction which exhibits a substantial hemoglobin concentration dependence over the concentration range 0-12 mg/mL. An equilibrium binding model yields a hemoglobin-membrane dissociation constant, Kd, on the order of 10(-4) M, at and above physiological pH; the interaction is classified as very low-affinity binding. The interaction increases significantly when the pH is decreased. Half-saturation of the binding sites occurs at a ratio of about 10(8) hemoglobins per cell. PMID- 6274391 TI - Polypeptide molecular weights of the (Na+,K+)-ATPase from porcine kidney medulla. AB - The molecular weights of the polypeptide chains from (Na+,K+)-ATPase of porcine kidney medulla have been determined by analytical sedimentation equilibrium. The alpha-subunit molecular weight is 93 900, and the beta subunit is a glycoprotein with a polypeptide molecular weight of 32 300 (41 400 including protein and carbohydrate). Amino acid and carbohydrate compositions are presented together with related properties (i.e., partial specific volumes, extinction coefficients, and hydrophobic/hydrophilic amino acid content). PMID- 6274392 TI - Phosphorus-31 nuclear magnetic resonance of phosphoenzymes of sodium- and potassium-activated and of calcium-activated adenosinetriphosphatase. AB - Prior studies identified phosphoenzyme intermediates in the turnover of sodium- and potassium-activated adenosinetriphosphatase [(Na,K)ATPase] from several sources and of the calcium-activated adenosinetriphosphatase [(Ca)-ATPase] of skeletal muscle sarcoplasmic reticulum. In both cases, the transphosphorylation is to a beta-aspartyl carboxyl group at the active site. We now report observation of a K+-sensitive phosphorylated intermediate of purified (Na, K)ATPase from the salt gland of the duck using high-field 31P nuclear magnetic resonance. Addition of ATP to a suspension of this enzyme in the presence of Mg2+ and Na+ produced a resonance at about +17 ppm relative to 85% phosphoric acid. Addition of inorganic phosphate and Mg2+ to (Na,K)ATPase also produced a resonance at about +17 ppm which was enhanced in the presence of a saturating concentration of the inhibitor, ouabain; again, addition of K+ made this resonance disappear. These findings are consistent with earlier kinetic characterization of an acid-stable (Na,K)ATPase phosphoenzyme intermediate by 32P labeled phosphate incorporation into a denatured precipitate of the enzyme. We attribute the +17-ppm resonance to formation of an acyl phosphate at an aspartyl residue of the catalytic site of (Na,K)ATPase. This is supported by our finding of a similar resonance at +17 ppm after phosphorylation of another membrane-bound cation transport enzyme, sarcoplasmic reticulum (Ca)ATPase, as well as by a similar resonance at about +17 ppm after phosphorylation of the model dipeptide L seryl-L-aspartate. PMID- 6274393 TI - Sulfur-free parathyroid hormone analogues containing D-amino acids: biological properties in vitro and in vivo. AB - Three sulfur-free analogues of bovine parathyroid hormone (bPTH) containing D amino acids were synthesized by the solid-phase method and their biological properties compared in an in vitro bioassay (rat renal adenylate cyclase assay), a receptor assay for parathyroid hormone (PTH) (canine renal membranes), and an in vivo bioassay (chick hypercalcemia assay). The analogue [Nle8,Nle18,D-Tyr34] bPTH-(1-34)-amide, which was found to be more than 4 times as potent in vitro as unsubstituted PTH, is the most potent analogue of PTH yet synthesized. The enhanced potency was largely attributable to increased affinity for the PTH receptor. In vivo, however, this analogue was only one-third as potent as bPTH-(1 34). Cumulative evidence suggests that the nearly 15-fold decline in the relative potency when the compound was assayed in vivo is due to the substitution of norleucine for methionine. The other analogues, [D-Val2,Nle8,D-Tyr34]bPTH-(1-34) amide and [D-Val2,Nle8,Nle18,D=Tyr34]bPTH-(2-34)-amide, were only weakly active in vitro and in vivo, indicating that substitution with D-amino acids at the NH2 terminus of PTH causes markedly diminished receptor affinity. In fact, the placement of a D-amino acid at the NH2 terminus is more deleterious to biological activity than is omission of amino acids at positions 1 and 2. PMID- 6274394 TI - Two contiguous thrombin fragments of human somatotropin form a functionally active recombinant, but the two homologous fragments from sheep hormone do not. AB - Two thrombin fragments of reduced-carbamidomethylated human somatotropin representing the full primary structure of the native hormone (residues 1-134 and 135-191) have been found to form a recombinant molecule with properties similar to those of reduced-carbamidomethylated human somatotropin as shown by circular dichroism spectroscopy, two receptor-binding assays, and radioimmunoassay. In contrast, the homologous thrombin fragments of reduced-carbamidomethylated sheep hormone (residues 1-133 and 134-191) do not undergo recombination. Furthermore, neither the reduced-alkylated nor the reduced and nonalkylated C-terminal thrombin fragment of sheep hormone is able to interact with the reduced carbamidomethylated N-terminal thrombin fragment of human hormone, under conditions which favor the recombination of the two human somatotropin fragments. PMID- 6274395 TI - Stoichiometry and spectral properties of the MoFe cofactor and noncofactor redox centers in the MoFe protein of nitrogenase from Azotobacter vinelandii. AB - Reductive EPR and optical titrations of oxidized MoFe protein using reduced methyl viologen as reductant were used to quantitate the stoichiometry of the various spectroscopically and electrochemically distinct redox centers in the oxidized MoFe protein. Three centers were found to correlate with the EPR signal development (MoFe cofactor centers), and three centers were found to be independent of the EPR signal (P clusters) but to demonstrate distinct optical and kinetic properties. Oxidative EPR and optical titrations of reduced MoFe protein are reported which support the presence of three P-cluster centers. The optical titrations show a distinct change in kinetic behavior between the MoFe cofactor and P-cluster centers. Controlled potential coulometry demonstrates that incremental oxidation of reduced protein by methylene blue, thionine, or indigodisulfonate occurs specifically at three P-cluster sites. Subsequent oxidation by methylene blue and thionine (but not indigodisulfonate) causes the EPR signal to disappear. Three P-cluster sites, two EPR sites, and one presently uncharacterized site are suggested by the results of this study. PMID- 6274396 TI - Membrane channel forming polypeptides. 270-MHz proton magnetic resonance studies of the aggregation of the 11-21 fragment of suzukacillin in organic solvents. AB - 270-MHz 1H NMR studies of the 11-21 suzukacillin fragment Boc-Gln-Aib-Leu-Aib-Gly Leu-Aib-Pro-Val-Aib-Aib-OMe (11-G) and its analogue Boc-Ala-Aib-Leu-Aib-Gly-Leu Aib-Pro-Val-Aib-Aib-OMe (11-A) have been carried out in CDCl3 and (CD3)2SO. The NH chemical shifts and their temperature coefficients have been measured as a function of peptide concentration in both solvents. It is established that replacement of Gln by Ala is without effect on backbone conformation. Both peptides adopt highly folded 310 helical conformations stabilized by seven intramolecular 4 leads to hydrogen bonds. Nonlinear temperature dependences are demonstrated for free NH groups in the Gln(1) peptide. Aggregation is mediated by intermolecular hydrogen bonds formed by solvent-exposed NH groups. A major role for the Gln side chain in peptide association is suggested by differences in the NMR behavior of the Gln(1) and Ala(1) peptides. For the Gln(1) peptide in CDCl3, the carboxamide side chain carbonyl group forms an intramolecular hydrogen bond to the peptide backbone, while the trans side chain NH shows evidence for intermolecular interactions. In (CD3)2SO, the cis carboxamide NH is involved in intermolecular hydrogen bonding. The possible role of the central Gln residue in stabilizing aggregates of peptide channel formers is discussed, and a model for hexameric association is postulated. PMID- 6274397 TI - Secondary structure of prokaryotic 5S ribosomal ribonucleic acids: a study with ribonucleases. AB - The structures of 5S ribosomal RNAs from Escherichia coli and Bacillus stearothermophilus were examined by using ribonucleases A, T1, and T2 and a double helix specific cobra venom ribonuclease. By using both 5' and 3'-32P-end labeling methods and selecting for digested but intact 5S RNA molecules, we were able to distinguish between primary and secondary cutting positions and also to establish the relative degree of cutting. The data reveal the predicted similarities of the higher order structure in the two RNAs but also demonstrate a few significant differences. The data also provide direct evidence for three of the helical regions of the Fox and Woese model of 5S RNA [Fox, G. E., & Woese, C. (1975) Nature (London) 256, 505] and support other important structural features which include a nucleotide looped out from a helical region which has been proposed as a recognition site for protein L18. PMID- 6274398 TI - Myxothiazol, a new inhibitor of the cytochrome b-c1 segment of th respiratory chain. AB - Myxothiazol inhibited oxygen consumption of beef heart mitochondria in the presence and absence of 2,4-dinitrophenol, as well as NADH oxidation by submitochondrial particles. The doses required for 50% inhibition were 0.58 mol myxothiazol/mol cytochrome b for oxygen consumption of beef heart mitochondria, and 0.45 mol/mol cytochrome b for NADH oxidation by submitochondrial particles. Difference spectra with beef heart mitochondria and with cell suspensions of Saccharomyces cerevisiae revealed that myxothiazol blocked the electron transport within the cytochrome b-c1 segment of the respiratory chain. Myxothiazol induced a spectral change in cytochrome b which was different from and independent of the shift induced by antimycin. Myxothiazol did not give the extra reduction of cytochrome b typical for antimycin. Studies on the effect of mixtures of myxothiazol and antimycin on the inhibition of NADH oxidation indicated that the binding sites of the two inhibitors are not identical. PMID- 6274399 TI - The insect brain (Na+ + K+)-ATPase. Binding of ouabain in the hawk moth, Manduca sexta. AB - (1) A quantitative study has been made of the binding of ouabain to the (Na+ + K+)-ATPase in homogenates prepared from brain tissue of the hawk moth, Manduca sexta. The results have been compared to those obtained in bovine brain microsomes. (2) The insect brain (Na+ + K+)-ATPase will bind ouabain either in the presence of Mg2+ and Pi, ('Mg2+, Pi' conditions) or in the presence of Na+, Mg2+, and an adenine nucleotide ('nucleotide' conditions) as is the case for the bovine brain (Na+ + K+)-ATPase. The binding conditions did not alter the total number of receptor sites measured at high ouabain concentrations in either tissue. (3) Potassium ion decreases the affinity (increases the KD) of ouabain to the M. sexta brain (Na+ + K+)-ATPase under both binding conditions. However, ouabain binding is more sensitive to K+ inhibition under the nucleotide conditions. In bovine brain ouabain binding is equally sensitive to K+ inhibition under both conditions. (4) The enzyme-ouabain complex has a rate of dissociation that is 10-fold faster in the M. sexta preparation than in the bovine brain preparation. Because of this, the M. sexta (Na+ + K+)-ATPase has a higher KD for ouabain binding and is less sensitive to inhibition by ouabain than the bovine brain enzyme. (5) This data supports the hypothesis that two different conformational states of the M. sexta (Na+ + K+)-ATPase can bind ouabain. PMID- 6274400 TI - Anomalous potential response and (Na+ + K+)-ATPase in in vitro frog gastric mucosa. AB - In general, increasing K+ on the nutrient side decreases the transmucosal PD (nutrient becomes more negative) but after bathing the mucosa in zero K+ media for about 30 min, or longer, elevation of K+ on the nutrient side increases the PD, an anomalous effect. In Cl- media, increasing nutrient K+ from zero to 4 mM produces an increase in PD (an anomalous response) of 3.1 and 5.3 mV in 2 and 5 min, respectively. Ouabain (10(-3) M) to the nutrient side abolished the anomalous response as did removal of Na+ (choline for Na+) from bathing media. In SO4(2-) media (SO4(2-) for Cl-), a significant anomalous PD response was observed when K+ on the nutrient side was increased from zero to 1, 2 or 3 mM but not to higher K+ concentrations. In this case, ouabain also abolished the anomalous response. It is postulated, on the basis of the effects of ouabain and the use of choline media, that an electrogenic (Na+ + K+)-ATPase pump is present on the nutrient-facing membrane in which more Na+ than K+ are transported per cycle. PMID- 6274401 TI - Studies on (Na+ + K+)-activated ATPase. XLV. Magnesium induces two low-affinity non-phosphorylating nucleotide binding sites per molecule. PMID- 6274402 TI - Studies on (Na+ + K+)-activated ATPase. XLIX. Content and role of cholesterol and other neutral lipids in highly purified rabbit kidney enzyme preparation. AB - (1) The neutral lipids and the free and bound fatty acids of a highly purified (Na+ + K+)-ATPase preparation from rabbit kidney outer medulla have been analysed. (2) On a dry weight basis, the total lipid content is nearly the same as the total protein content, and consists for 66% of phospholipids and for 34% of neutral lipids and free fatty acids. In the latter category cholesterol is the main component (71%). (3) On a molar basis the enzyme preparation contains 382 mol phospholipids, 67 mol free fatty acids, 9, 16 and 12 mol mono-, di- and triacylglycerols, 249 and 19 mol free and esterified cholesterol per mol enzyme. (4) The fatty acid composition of each lipid and of the free fatty acid fraction, present in the enzyme preparation, is reported. (5) All cholesterol and part of the phospholipids can be removed by hexane extraction, leaving 66% of the (Na+ + K+)-ATPase activity. Oxidation of all cholesterol to cholest-4-en-3-one by cholesterol oxidase leaves 85% of the (Na+ + K+)-ATPase activity. These results indicate that cholesterol is not essential for (Na+ + K+)-ATPase activity. PMID- 6274403 TI - Relation between red cell membrane (Na+ + K+)-ATPase and band 3 protein. AB - This study is designed to examine the participation of the major red cell membrane protein, band 3 protein, in the chain which transmits information from the cardiac glycoside site on the external face of the cell (Na+ + K+)-ATPase to the megadalton glycolytic enzyme complex within the cell. The experiments show that the anion transport inhibitor, 4,4'-diisothiocyano-2,2'-stilbenedisulfonic acid, affects the resonance of 2,3-diphosphoglycerate, as does the cardiac glycoside cation transport inhibitor, ouabain. Resonance shifts induced by the cardiac glycoside alone are modulated by addition of the anion transport inhibitor which indicates that there is coupling in the red cell between the (Na+ + K+)-ATPase and band 3 protein. Band 3 protein was separated from the membrane and partially purified following the technique of Yu and Steck ((1975) J. Biol. Chem. 250, 9170-9175). When glyceraldehyde-3-phosphate dehydrogenase was added to the separated band 3 protein preparation, addition of cardiac glycosides caused shifts in the 31P resonance of glyceraldehyde 3-phosphate. These experiments indicate that there is coupling between the (Na+ + K+)-ATPase and band 3 protein in the separated preparation and suggest that the anion and cation transport systems may be closely related spatially and functionally in the intact red cell. PMID- 6274404 TI - Localization of 2',3'-cyclic nucleotide 3'-phosphodiesterase in human erythrocyte membranes. AB - The location of 2',3'-cyclic nucleotide 2',3'-phosphodiesterase in human erythrocyte membranes was determined. This was accomplished by comparing the enzyme's accessibility with that of glyceraldehyde-3-phosphate dehydrogenase (cytoplasmic surface marker) and acetylcholinesterase (external marker) in sealed and unsealed ghosts and normal and inverted membrane vesicles. The results showed that 2',3'-cyclic nucleotide 3'-phosphodiesterase, like glyceraldehyde-3 phosphate dehydrogenase, meets several criteria for an inner (cytoplasmic) membrane location: (1) the enzyme was accessible to substrate in unsealed ghosts and inside-out vesicles but not in sealed or right-side-out vesicles, (2) latent activity in sealed ghosts could be exposed with detergent (Triton X-100), (3) activity in unsealed ghosts was gradually sequestered during resealing and could be re-exposed with detergent, and (4) the enzyme was susceptible to trypsin proteolysis only in unsealed ghosts. These results demonstrate that the active site of 2',3'-cyclic nucleotide 3'-phosphodiesterase faces the cytoplasm of erythrocytes and that the enzyme may not span the lipid bilayer of the membrane. The localization of the phosphodiesterase on the inner membrane surface of erythrocytes suggests that the similar enzyme of myelin may be embedded within the major dense line of the compact lamellae. PMID- 6274405 TI - Purification and characterization of plasma membrane fractions from cultured pituitary glands. AB - Highly purified plasma membrane fractions have been prepared from GH3 pituitary cells grown in suspension cultures. These membrane fractions have been obtained by differential and sucrose gradient centrifugation and were characterized in terms of their lipid content, marker enzyme analysis and the binding of 3H labelled thyrotropin-releasing hormone (TRH) to its receptor. Alkaline phosphatase and 5'-nucleotidase activities were enriched 12-to 15-fold in the plasma membrane fraction with somewhat greater enrichment (28-fold) of the specific binding component for [3H]TRH, with a specific activity of 2286 fmol [3H]TRH bound per mg protein. A single class of binding sites for TRH was observed with an apparent dissociation constant of 18 nM, a value similar to that observed for intact cells. No detectable TRH binding to the nuclear fraction was observed that could not be ascribed to residual plasma membrane contamination. By electron microscopy, these fragments appeared to be sealed vesicles with an average diameter of approximately 1800 A. The binding of 125I-labelled wheat germ agglutinin was used as a marker for plasma membrane purification. In addition to specific binding to this membrane fraction, specific binding was also observed in the nuclear fraction. Studies with fluorescein-labelled wheat germ agglutinin revealed that, in fixed cells, fluorescence was restricted to the plasma membrane. However, if the cells were treated with Triton before labelling, most of the fluorescence was then associated with the cell nucleus. Hence, the use of wheat germ agglutinin binding as a specific membrane marker must be reevaluated. PMID- 6274406 TI - Chronic exposure to inhaled anesthetics increases cholesterol content in Acholeplasma laidlawii. AB - Acholeplasma laidlawii cells were grown in cholesterol-enriched medium and exposed continuously to either air (control), 4.0 vol.% halothane in air at 1 atm pressure (4% atm halothane), or 80% cyclopropane in oxygen for 24 h at 37 degrees C. Cells grown in the presence of 4% atm halothane or 80% cyclopropane had approximately twice as much membrane cholesterol content/mg protein as the control cells. Cells grown in an anesthetic environment also tended to have a higher membrane cholesterol/phospholipid molar ratio compared to control cells. Membranes isolated from halothane-exposed cells grown in a cholesterol-enriched medium were more ordered at 37 degrees C (measurements were made with no anesthetic present) than membranes from control cells grown in an identically enriched medium. This difference in membrane physical state between control and anesthetic-exposed cells decreased as the temperature decreased, and disappeared at approx. 23 degrees C. Continuous exposure of A. laidlawii to 4% atm halothane or 80% cyclopropane for 24 h did not markedly affect membrane fatty acid composition, either in cells grown on an unsupplemented medium or in cells grown in a medium enriched in myristic, palmitic or stearic acids. These results further support the hypothesis that an increased membrane cholesterol content may play a role in the tolerance or dependence that develops after chronic exposure to anesthetic agents. PMID- 6274407 TI - Comparison of the interaction of cyclic nucleotide-dependent protein kinases with mononucleosomes and free histones. AB - Arginine-rich histones H2A, H2B, H3 and H4 contain two regions of interaction with cyclic nucleotide-dependent protein kinases: a substrate phosphorylation site and a region which noncompetitively inhibits cyclic nucleotide binding to the protein kinases. We have compared the interaction of cyclic nucleotide dependent protein kinases with these two sites in histones which are organized in nucleosome structures with the interaction of the enzymes with free histones. Whereas histones in solution are readily phosphorylated by cyclic GMP-dependent protein kinase and the catalytic subunit of cyclic AMP-dependent protein kinase, mononucleosomes are not phosphorylated by these enzymes. Histones extracted from mononucleosomes can be phosphorylated, indicating that the lack of phosphorylation of nucleosomes is not due to covalent modification of the histones but to their organization within the nucleosome structure. Whereas histones in solution are effective noncompetitive inhibitors of cyclic GMP binding to cyclic GMP-dependent protein kinase and of cyclic AMP binding to the regulatory subunits of cyclic AMP-dependent protein kinase, mononucleosomes do not affect cyclic nucleotide binding. These studies indicate that histones which are organized in nucleosome structures are neither substrates nor modifiers of cyclic nucleotide-dependent protein kinases. PMID- 6274408 TI - Purification and properties of a neutral endodeoxyribonuclease from guinea pig epidermis. AB - Guinea pig epidermal DNAase I was purified from an epidermal extract by a procedure including DEAE-cellulose chromatography, Sephadex G-100 gel filtration and Con A-Sepharose affinity chromatography. The purified enzyme contained no detectable activities of acid DNAase, alkaline RNAase, phosphodiesterase or acid or alkaline phosphatase, but was contaminated with acid RNAase activity. The molecular weight of the enzyme was estimated to be 33 000 by sucrose density gradient centrifugation and Sephadex G-100 gel filtration. Its isoelectric point is 5.2 +/- 0.1. The enzyme requires divalent cations and exhibits two pH optima that are dependent on divalent cations: in the presence of Mn2+, the optimum pH is about 7.5 in 50 mM Tris-HCl buffer and in the presence of Mn2+, the pH is 6.4 in 50 mM cacodylate-HCl buffer. The enzyme hydrolyzes native DNA about 6-times faster than denatured DNA, producing 5'-phosphoryl and 3'-hydroxyl terminated oligonucleotides with an average chain length of about eight nucleotides, and converts double-stranded and circular DNA to relaxed and linear forms. The enzyme is inhibited by G-actin and antiserum against bovine pancreatic DNAase A. Thus this enzyme is classified as DNAase I. PMID- 6274409 TI - In vitro conversion of S13 viral DNA in phage particles to the double-stranded DNA. AB - The conversion of single-stranded DNA in S13 intact phage particles to the double stranded replicative form DNA was observed in cell extracts prepared from Escherichia coli H560 (S13s, polA, endA) cells lysed with lysozyme and the non ionic detergent, Brij 58. The DNA product, which associated with a rapidly sedimenting component, was identified as RFII-DNA with a gap by sedimentation analysis. The conversion was inhibited by N-ethylmaleimide, but not by rifampicin, nicotinamide mononucleotide or polymyxin B. The dnaB gene product was involved in the replicative system. Similar extracts prepared from a S13 resistant E. coli strain K12W6 also catalyzed this synthesis. PMID- 6274410 TI - S1 nuclease does not cleave DNA at single-base mis-matches. AB - Three assays have been designed to detect the cleavage of duplex phi X174 DNA at single-base mis-matches. Studies with S1 nuclease failed to detect cleavage at mis-matches. S1 nuclease digestion at 37 and 55 degrees C failed to produce a preferential degradation of a multiply mis-matched heteroduplex when compared to a mis-match-free homo-duplex as analyzed by sedimentation on sucrose gradients. Other heteroduplex templates were not cleaved by S1 nuclease at a defined single base mis-match when assayed by gel electrophoresis or by marker rescue. In all cases, the amount of S1 nuclease employed was at least 10-times more than that required to render a single-stranded phi X174 DNA molecule completely acid soluble. The rate of hydrolysis of single-base mis-matches by S1 nuclease was estimated to be less than 0.016% of the rate at a base in single-strand phi X174 DNA. In no instance did we detect activity by S1 nuclease directed at mis-matched sites in our template molecules. Similarly, the single-strand specific endonuclease from Neurospora crassa does not cleave heteroduplex templates at a defined single-base mis-match when assayed by marker rescue. PMID- 6274411 TI - In vitro inhibition of yeast valyl-tRNA synthetase by the valine homologue of ochratoxin A. PMID- 6274412 TI - The kinetics of unphosphorylated, phosphorylated and proteolytically modified fructose bisphosphatase from fat liver. AB - Phosphorylation of fructose-bisphosphatase (D-fructose-1,6-bisphosphate 1 phosphohydrolase, EC 3.1.3.11) by the catalytic subunit of cyclic AMP-dependent protein kinase from pig muscle decreased the K0.5 for fructose-bisphosphate from 21 to 11 microM. When the phosphorylated fructose-bisphosphatase was treated with trypsin the K0.5 increased to 22 microM. The K0.5 also increased when the phosphoenzyme was treated with a partially purified phosphatase from rat liver. There was no difference between the unphosphorylated and phosphorylated enzyme with respect to pH dependence, the pH optimum being about 7.0 for both. Limited treatment of fructose-bis-phosphatase with subtilisin, which cleaves the enzyme at its unphosphorylatable N-terminal part, increased the pH optimum more than limited treatment with trypsin, which releases the phosphorylated peptide at the C-terminal part of fructose-bisphosphatase. The phosphorylated site on the phosphorylated fructose-bisphosphatase was more easily split off by trypsin treatment than the corresponding unphosphorylated site. The results suggest in addition to the glucagon-induced phosphorylation of fructose-bisphosphatase described by Claus et al. [1] that the phosphorylation-dephosphorylation of fructose-bisphosphatase could be of importance for the hormonal regulation of the enzyme in vivo. PMID- 6274413 TI - Tripeptidyl carboxypeptidase activity of kininase II (angiotensin-converting enzyme). AB - The degradation of des-Arg9-brady kinin and its analogues by highly purified preparations of hog lung and kidney kininase II (angiotensin-converting enzyme; peptidyldipeptide hydrolase, EC 3.4.15.1) was studied. The degradative peptides fragments were separated and isolated by high performance liquid chromatography and identified by amino acid analysis. Both enzymes released C-terminal tripeptides from des-Arg9-bradykinin, des-Arg9-(Leu8)-bradykinin, Pro-Pro-Gly-Phe Ser-Pro-Phe, Pro-Gly-Phe-Ser-Pro-Phe, Gly-Phe-Ser-Pro-Phe, Bz-Gly-Ser-pro-Phe and Bz-Gly-Ala-Pro-Phe. Hydrolysis of Phe-Ser-Pro-Phe, Bz-Gly-His-Pro-Phe, Bz-Gly-Phe Pro-Phe and Bz-Gly-Gly-Pro-Phe by both enzymes was negligible. These data indicate that kininase II can release C-terminal tripeptides of substrates having a proline residue in the penultimate position such as des-Arg9-bradykinin and its analogues, and that this enzyme is able not only to act as a dipeptidyl carboxypeptidase but also acts as a tripeptidyl carboxy-peptidase. The tripeptidyl carboxypeptidase enzyme was sensitive to inhibition by kininase II inhibitors. PMID- 6274414 TI - Identification of platelet receptors for bovine von Willebrand factor on human platelets by a new platelet receptor test. AB - A new test for the quantitation of platelet receptor activity for bovine von Willebrand factor in human platelet extracts is described. The test is based on the competition for the von Willebrand factor occurring between platelets, an antibody against von Willebrand factor, and platelet extracts containing the von Willebrand receptor. Bovine von Willebrand factor aggregates human platelets directly. In the presence of EDTA, anti-bovine von Willebrand factor antibody disaggregates platelet aggregates induced by bovine von Willebrand factor. Disaggregation is measured by following the decrease in light transmission accompanying disaggregation. Platelet extracts inhibited the disaggregating activity of the antibody, and the percent inhibition was directly proportional to the amount of the platelet extract added. The decrease in platelet disaggregating activity of the antibody was used as an indicator of platelet receptor activity in a system consisting of platelet-rich plasma, bovine von Willebrand factor, antibody and platelet extract. With this method we found von Willebrand factor receptors in the whole platelet extract, the Triton X-100 platelet membrane extract and in platelet extract material which bound to a bovine Willebrand factor-Sepharose 6MB affinity column. PMID- 6274415 TI - Differences in the beta-adrenergic responsiveness between high and low passage rat glioma C6 cells. AB - Responsiveness to catecholamines was studied in two different strains of rat glioma C6 cells. The C6 cells of low passage possessed a high capacity to accumulate cyclic AMP in response to (-)-isoproterenol. Cholera toxin was also able to stimulate cyclic AMP accumulation in these cells. High passage C6 cells were unresponsive to (-)-isoproterenol or to cholera toxin except in the presence of a high concentration of phosphodiesterase inhibitor. The affinity of beta adrenergic receptors on both strains for (-) [3H] dihydroalprenolol was similar; however, C6 low passage possessed several times the number of beta-adrenergic receptors found in C6 high passage. This difference correlated with the difference found in (-)-isoproterenol-stimulated adenylate cyclase between C6 low passage and high passage. The sodium fluoride-stimulated adenylate cyclase was similar in both strains. Cyclic AMP phosphodiesterase activity was 2-3 times higher in homogenates of C6 high passage than in low passage. In intact cells, the rate of breakdown of cyclic AMP was 5-times faster in C6 high passage than in low passage. Thus, differences in beta-adrenergic receptor number and phosphodiesterase activity explain in part the lack of responsiveness of C6 high passage. Our studies indicate that continuous subculturing of rat glioma C6 cells led to complex alterations in the beta-adrenergic receptor-adenylate cyclase system. PMID- 6274416 TI - Activation of superoxide production and differential exocytosis in polymorphonuclear leukocytes by cytochalasins A, B, C, D and E. Effects of various ions. AB - All of the common cytochalasins activate superoxide anion release and exocytosis of beta-N-acetylglucosaminidase and lysozyme from guinea-pig polymorphonuclear leukocytes (neutrophils) incubated in a buffered sucrose medium. Half-maximal activation of both processes is produced by approx. 0.2 microM cytochalasin A, C greater than 2 microM cytochalasin B greater than or equal to 4-5 microM cytochalasin D, E. While maximal rates of O2- release and extents of exocytosis require extracellular calcium (1-2 mM), replacing sucrose with monovalent cation chlorides is inhibitory to neutrophil activation by cytochalasins. Na+, K+ or choline inhibit either cytochalasin B- or E-stimulated O2- production with IC50 values of 5-10 mM and inhibition occurs whether Cl-, NO3- or SCN- is the anion added with Na+ or K+. Release of beta-N-acetylglucosaminidase in control or cytochalasin B-stimulated cells is inhibited by NaCl(IC50 approximately 10 mM), while cytochalasin E-stimulated exocytosis is reduced less and K+ or choline chloride are ineffective in inhibiting either cytochalasin B- or E-stimulated exocytosis. Release of beta-glucuronidase, myeloperoxidase or acid phosphatase from neutrophils incubated in buffered sucrose is not stimulated by cytochalasin B. Stimulation of either O2- or beta-N-acetylglucosaminidase release by low concentrations of cytochalasin A is followed by inhibition of each at higher concentrations. It appears that all cytochalasins can activate both NAD(P)H oxidase and selective degranulation of neutrophils incubated in salt-restricted media and that differential inhibition of these two processes by monovalent cations and/or anions is produced at some step(s) subsequent to cytochalasin interaction with the cell. PMID- 6274417 TI - Effect of thyroid status on alpha- and beta-catecholamine responsiveness of hamster adipocytes. AB - It has been suggested that part of the increased beta-catecholamine responsiveness in hyperthyroid animals is due to a decrease in alpha catecholamine action. The present results indicate that neither hyperthyroidism nor hypothyroidism altered the alpha 2-adrenergic inhibition of adenylate cyclase or the alpha 1-adrenergic stimulation of phosphatidylinositol turnover in adipocytes from the white adipose tissue of hamsters. No effect of hyperthyroidism was found on the Kd for binding of [3H]dihydroergocryptine or the number of binding sites in membranes prepared from hamster adipocyte tissue. The stimulation of cyclic AMP due to beta-catecholamines was enhanced in adipocytes from hyperthyroid hamsters, as was lipolysis. However, in adipocytes from hyperthyroid hamsters the maximal stimulation of cyclic AMP due to isoproterenol, ACTH or epinephrine plus yohimbine, as seen in the presence of adenosine deaminase and theophylline, was less than in adipocytes from euthyroid hamsters. The activation of adenylate cyclase by isoproterenol was the same in membranes from hyperthyroid as compared to those from euthyroid hamsters in the absence or presence of guanine nucleotides. These data suggest that thyroid status has little effect on alpha-catecholamine action by enhances the activation of lipolysis by beta-catecholamine agonists. PMID- 6274418 TI - Cyclic nucleotides in the denervated rat diaphragm and the effect of cyclic AMP on ornithine and adenosylmethionine decarboxylases. AB - The concentrations of cyclic AMP and cyclic GMP were measured in the denervated rat diaphragm at various times following unilateral phrenicectomy. Cyclic AMP concentration was raised by the second day after operation, reached a peak by the third day, followed by another increase at around 10 days. By contrast, cyclic GMP concentration was decreased within a day after denervation and remained below control levels at all subsequent times studied. Epinephrine in vitro produced a comparable increase in the concentration of cyclic AMP in both normal and denervated tissue. The concentration of adenosine appeared unchanged in the denervated diaphragm by comparison with its innervated control. Activity of ornithine decarboxylase was elevated in the diaphragms of rats treated with dibutyryl cyclic AMP, but this effect could also be achieved with sodium butyrate alone. Adenosylmethionine decarboxylase activity, was unaffected after treatment with either compound. These observations and others discussed are taken to indicate a lack of direct relationship between cyclic AMP concentrations and the activity of the rate-limiting enzymes of polyamine biosynthesis in the rat diaphragm. PMID- 6274419 TI - Induction and repression of enzymes involved in exogenous purine compound utilization of Bacillus cereus. AB - 5'-Nucleotidase, adenosine phosphorylase, adenosine deaminase and purine nucleoside phosphorylase, four enzymes involved in the utilization of exogenous compounds in Bacillus cereus, were measured in extracts of this organism grown in different conditions. It was found that adenosine deaminase is inducible by addition of adenine derivatives to the growth medium, and purine, nucleoside phosphorylase by metabolizable purine and pyrimidine ribonucleosides. Adenosine deaminase is repressed by inosine, while both enzymes are repressed by glucose. Evidence is presented that during growth of B. cereus in the presence of AMP, the concerted action of 5'-nucleotidase and adenosine phosphorylase, two constitutive enzymes, leads to formation of adenine, and thereby to induction of adenosine deaminase. The ionsine formed would then cause induction of the purine nucleoside phosphorylase and repression of the deaminase. Taken together with our previous findings showing that purine nucleoside phosphorylase of B. cereus acts as a translocase of the ribose moiety of inosine inside the cell (Mura, U., Sgarrella, F. and Ipata, P.L. (1978) J. Biol Chem. 253, 7905-7909), our results provide a clear picture of the molecular events leading to the utilization of the sugar moiety of exogenous AMP, adenosine and inosine as an energy source. PMID- 6274421 TI - Vanadate: a potent inhibitor of multifunctional glucose-6-phosphatase. AB - Vanadate has been found to be a potent inhibitor of both the hydrolytic and synthetic activities of the multifunctional enzyme glucose-6-phosphatase (D glucose-6-phosphate phosphohydrolase, EC 3.1.3.9). The enzyme, when studied in both microsomal preparations and in situ using permeable isolated hepatocytes, is inhibited by micromolar concentrations of vanadate. The inhibition by vanadate is greater in detergent-treated than in untreated microsomes. In both the microsomal preparations and permeable hepatocytes, the inhibition by vanadate is competitive with the phosphate substrate and is greater for the phosphotransferase than the hydrolase activity of the enzyme. The Ki values of vanadate for carbamyl phosphate : glucose phosphotransferase and glucose-6-phosphate phosphohydrolase determined with permeable hepatocytes are in good agreement with the values determined with detergent-dispersed microsomes. The previously described inhibition of glucose-6-phosphate phosphohydrolase by ATP (Nordlie, R.C., Hanson, T.L., Johns, P.T. and Lygre, D.G. (1968) Proc. Natl. Acad. Sci. USA 60, 590-597) can now be explained by the vanadium contamination of the commercially available ATP samples used. In contrast with glucose-6-phosphatase, hepatic glucokinase and hexokinase were not inhibited by vanadate. Physiological implications and utilitarian experimental applicability of vanadate as a selective metabolic probe, based on these observations, are suggested. PMID- 6274420 TI - Platelet aggregation and sphingomyelinase D activity of a purified toxin from the venom of Loxosceles reclusa. AB - A facile and quantitative assay for measuring the activity of sphingomyelinase D in recluse spider venom has been developed using L-alpha-[palmitoyl-1 14C]lysophosphatidylcholine as substrate. This assay avoids the problem of substrate insolubility that occurs when sphingomyelin and other insoluble lipids are used as substrates. This assay has been employed in gel filtration and isoelectric focusing isolation techniques to purify sphingomyelinase D from spider venom. The purified sphingomyelinase exhibits four active enzyme forms in isoelectric focusing with pI values of 8.7, 8.4, 8.2, and 7.8. Each active form when examined in SDS-polyacrylamide gel electrophoresis gave an estimated molecular weight of 32 000. The four active enzyme forms were immunologically cross-reactive with each other as demonstrated with radioimmune assays using an antiserum developed to one of the active forms. Each active form hydrolysed sphingomyelin to release choline and produce N-acylsphingosine phosphate. One of the active enzyme forms was characterized further in dermonecrosis and platelet aggregation measurements. This purified sphingomyelinase D was identified as a poisonous toxin that can developed typical dermonecrotic spider lesions when injected into experimental animals at levels expected to be delivered in a normal bite. Furthermore, the purified toxin acts to aggregate human blood platelets. The toxin-induced platelet aggregation has been related to serotonin release as aggregation occurs, and it has been shown to be inhibited by EDTA over the range of 0.6 yo 3.0 mM EDTA. It is suggested that spider-induced dermonecrosis could result in part from platelet aggregation at and near the site of envenomation. PMID- 6274422 TI - [Functions of pheophytin, plastoquinone, iron and carotenoids in plant photosystem 2 reaction centers]. AB - Photoreduction of the intermediary electron acceptor, pheophytin (Ph), in photosystem-2 (PS-2) reaction centers of spinach chloroplasts or subchloroplast particles (TSF-II and TSF-IIa) at 220 K and Eh approximately -450 mV produces a narrow ESR signal of Ph. (g = 2.0033; delta H approximately 13 G) and a "doublet" centered at g = 2.00 with a splitting of 52 G at 7 K. The doublet (but not the narrow signal) is eliminated after extraction of lyophylized TSF-II with hexane, containing 0.1-0.2% methanol, or after extraction of Fe with LiClO4 and o phenantroline, and the signal is restored by reconstitution with plastoquinone-A (PQ) or Fe++, respectively. The Fe removal results also in the development of a photoinduced ESR signal of PQ. (g approximately 2.0044; delta H approximately 9.2 G). The conclusion is made that the primary electron acceptor, Q, is in fact a complex PQ-Fe++ and that the exchange interaction of Ph. with PQ. -Fe++ is responsible for the ESR doublet. Photoreduction of Ph in TSF-IIa is accompanied by the 3-fold decrease in the formation of carotenoid triplet state (measured by the characteristic flash-induced absorbance changes) which is suggested to be a result of charge recombination in the pair [P680+ .PH.]. PMID- 6274423 TI - [Breakdown of guanosine-5'-monophosphate by UV- and gamma-irradiation in a vacuum]. PMID- 6274424 TI - [Effect of polymyxin B on the conductivity of negatively charged bilayer lipid membranes]. AB - Effect of polymyxin B on the planar bilayer lipid membranes (BLM) formed from synthetic phosphatidic acid has been studied. The addition of cholesterol to phospholipid in molar ratio 1 : 2 was followed by an increase of BLM conductance from 2 x 10(-8) to 3 x 10(-7) Ohm-1 cm-2. It was suggested that the observed increase of conductance was due to the fluidity of the membrane matrix in the presence of cholesterol. It was shown that 10(-6)--10(-5) M polymyxin slightly affected the conductance of BLM from phosphatidic acid. It was found that polymyxin increased conductance of negatively charged BLM modified by palmitic acid from 10(-8) to 10(-6) Ohm-1 cm-2. PMID- 6274425 TI - [The reaction between myosin and a spin-labeled derivative of ATP and Mn2+ at room temperature]. PMID- 6274426 TI - [Paramagnetic label study of actin]. PMID- 6274427 TI - [Localization of metallo-acceptor centers specifically binding divalent cations in myosin molecules]. AB - The effects of DTNB and 4 M LiCl on ESR spectra saturation curves of myosin, spin labelled at S1-SH groups in the presence of low concentrations of Mg2+ and Mn2+ ions have been studied. It has been shown that both DTNB and 4 M LiCl have taken off the influence of Mn2+ bound to high affinity metal binding sites of myosin on the character of saturation curves of spin-labelled protein. The results lead to a conclusion that high affinity metal binding sites of myosin are located in the LC2.light chains. PMID- 6274428 TI - [Light-induced hyperpolarization of membrane potential in Nitella flexilis call plasmalemma]. AB - Light-induced hyperpolarization of the membrane potential in Nitella flexilis cell plasmalemma was investigated by the clamping method. It is shown that this response is of an excitation character. The equivalent electric diagram used in the work and the electromotive force included on its basis allowed consideration of two possible (from the common point of view) mechanisms of the response: the action of the electrogenic ion pump and membrane specific permeability to a definite ion. The membrane being in the hyperpolarization state is not capable of acting as a bicarbonate electrode. It is suggested that the response under study is defined by the action of the electrogenic pump, the role of which is performed by HCO2(3)-dependent H+-ATPase with the ion channel as an exciting one. PMID- 6274429 TI - [Detection of the lethal process in plankton noctiluca by means of a forbidden transition of ESR of Mn2+ ion]. AB - A forbidden ESR line of Mn2+ that is connected with the penetration of Mn into the plancton organism and binding it to a marcomolecule is selected from the experiment. A method for saturating the plancton organism with paramagnetic ions is proposed. It is shown that the constant of the axial electric field in the spin hamiltonian of Mn2+ ion described the dynamics of a selforganizing system. It is tested that the lethal process in the plancton with paramagnetic ion enrichment originated from boson avalanche. Experiments are performed with plancton noctiluca which illustrate the occurrence of avalancheline lethal process in the case of paramagnetic ion enrichment with limiting concentration. The meaning of these results for the problems of oceanology and pollution-ocean inhabitants interaction in the case of paramagnetic ions is discussed. PMID- 6274430 TI - [Intracellular calcium-initiator of inactivation of choline receptors]. AB - It has been shown that inactivation of cholinoreceptors observed in the region of neuron depolarization is conditioned by the supply of Ca ions into the cell through the electro--excitable Ca2+-channels. It is proved by three means: 1) variation of outer concentration of Ca2+(Ca2+h); 2) application of the blockers of electroexcitable Ca2+ channels; 3) variation of intracellular concentration Ca2+(Ca2+b). PMID- 6274431 TI - [Mechanism of activation of Na,K-ATPase in nerve fibers during rhythmic excitation]. AB - The activation mechanism of Na,K-ATPase in nerve fibres after rhythmic excitation was studied. 3H-ouabain binding to a nerve was found to depend on the frequency of rhythmic excitation. The maximum of 3H-ouabain binding to a nerve crab was at 10 imp/sec. Rhythmic excitation was found not to change Na,K-ATPase affinity to ouabain, but appeared to increase the concentration of ouabain-sensitive sites in the nerve membrane. Transformation of inactive forms of the enzyme into active ones was supposed to be a possible cause of greater 3H-ouabain binding to the nerve during rhythmic exitation. PMID- 6274432 TI - [Effect of polymyxin B on ion permeability of bacterial membranes]. AB - Effect of polymyxin B on the movement of K+ and H+ in polymyxin-sensitive cells of E. coli under different metabolic states has been studied. It was shown that polymyxin B induced the efflux of K+, decreased the efflux of H+ and inhibited the consumption of oxygen in bacterial cells. The effect of antibiotic on ion movement was independent of respiratory conditions. It was suggested that polymyxin B increased ion permeability and destroyed lipid-protein interactions of the respiratory chain simultaneously. PMID- 6274433 TI - [Proton generator of superhigh frequency]. AB - Possible mechanism of superhigh frequency (10(10)-10(12) Hz) electromagnetic oscillation generation by an external proton in a system of hydrogen bonds of biomacromolecules is briefly discussed. The external proton in a proton channel deforms the potential profile of the proton of hydrogen bond in such a way, that there appears a possibility of the low frequency proton tunneling along the hydrogen bond. The interaction with the neighbouring bonds leads to further lowering of the generated frequency. PMID- 6274434 TI - [Analysis of ESR spectra in Mn2+-plant adenylate kinase complex]. AB - Interaction of plant adenylate kinase with Mn2+-adenine nucleotide binary complex was studied by ESR technique at room temperature. The ligand environment of Mn2+ in the ternary Mn2+-adenine nucleotide-enzyme complex was shown to change, as a result of enzyme binding as compared with that of binary complex. These changes seem to be due to substitution of protein molecules for water and adenine nucleotide ones, coordinated to Mn2+ ion on ternary complex formation. The same results were obtained in ESR studies on rabbit muscle myokinase. This fact may be considered as an evidence, that plant adenylate kinase is identical to animal one in its interaction with adenine nucleotides and manganese ions. PMID- 6274435 TI - [Selectivity of the channels formed by flavomycoin in a lipid bilayer as a function of the concentrations of transported ions]. AB - Experiments on selective properties of the flavomycoin channels in a lipid bilayer in the solution of KCl show that the permeability ratio PCl/PK decreased with KCl concentration. A mathematical model of ion transport is developed which provides a quantitative description of the observed nonmonotonic dependence of transmembrane potential at a zero current on the concentration of KCl. An increase in PK relative to PCl is due to the difference in location of the binding sites of K+ and Cl- in a channel. The binding sites of Cl- are closer to the channel mouth as compared with those of K+. PMID- 6274437 TI - [Light-induced generation of membrane potential in intact chloroplasts suspended in H2O or D2O media]. AB - The membrane potential changes induced by short flashes and continuous light were investigated in isolated chloroplasts of Peperomia metallica suspended in H2O- or D2O media. The potential generated in H2O-suspended chloroplasts by a single turnover flash is approximately two times lower than the maximal level of potential induced by continuous light. The photoelectric response of D2O suspended chloroplasts differs from that of H2O-suspended chloroplasts by an increased amplitude and a prolonged phase of the potential rise. Te dark decay of the potential proceeds 2-3 times slower in the D2O-suspended chloroplasts as compared to the H2O-suspended chloroplasts. The magnitude of the flash-induced potential is somewhat lower for the chloroplasts in D2O than for the chloroplasts in the H2O medium. The results obtained suggest that the substitution of H2O for D2O results in a decrease of the ionic conductance and an increase of stability of thylakoid membranes. It was shown that the rise of electrical potential under continuous illumination proceeds in two stages. The difference kinetics of membrane potential changes are observed under conditions of separate activity of two systems of photosynthesis. PMID- 6274436 TI - [Sites responsible for stimulation of brain Na,K-ATPase by serotonin]. AB - Serotonin (10(-6)-10(-3) M) stimulates Na,K-ATPase in the rat brain cortex homogenate with a maximal effect at 10(-4) M. Deseril (10(-5) M), antagonist of serotonin receptors, removes the stimulating effect. Deseril has no influence on noradrenaline-induced ATPase activation; the alpha-adrenergic blocker phentolamine does not affect serotonin activation. The effects of the two transmitters are additive. It is assumed that the mediators interact with different membrane sites, serotonin activation being initiated via the serotonin receptors. PMID- 6274438 TI - [Mapping of the 16S rRNA regions in ribosomes capable of complementary binding of oligonucleotides]. AB - 16S rRNA regions have been mapped on the surface of the 30S ribosomal subunit of E. coli due to their ability to bind the statistical mixture of hexadeoxyribonucleotides and thus to be hydrolyzed with RNAase H. These regions were found to be located around 80, 996-998, 1044-1046, 1408-1410, 1423, 1484 1485, 1495, 1500-1506, 1531-1532 16S rRNA nucleotide residues. PMID- 6274439 TI - Correlation between mixed lymphocyte cultures and anti-Epstein Barr virus (EBV) antibodies in Hodgkin's disease and non Hodgkin lymphoma patients. AB - Cellular and humoral immunity was examined in Hodgkin's disease and non Hodgkin lymphoma (33) patients and compared with a normal group. Mixed lymphocyte cultures (MLC) were used as parameter for cell mediated immunity and anti-Epstein Barr virus (EBV) antibodies for specific humoral immunity. High stimulation indices coincided with low anti-VCA and anti-EBNA titers in the control group (r = -0.343). This negative correlation was not found in non Hodgkin lymphomas and was substantially lower (r = -0.142) in Hodgkin's disease. The alterations in immunoregulatory mechanisms in these patients are discussed. PMID- 6274440 TI - Unmodified phagocytosis after periodate treatment of human granulocytes separated by a two-step method using Ficoll-Hypaque and polyvinylic alcohol medium. AB - A two-step method separating granulocytes and mononuclear cells from a single sample of 10 ml human blood is described. The technique combines a Ficoll-Hypaque treatment and a sedimentation in a Polyvinylic alcohol medium. This method (FHP) is then compared to the Ficoll-Hypaque-Dextran procedure. The FHP method requires approximately 2 h to be carried out and results in the preparation of granulocytes (greater than 99% purity) with high yield (40.7 % +/- 9.1). The cell viability exceeds 95%. Moreover the enzymic and phagocytic properties of polymorphonuclear cells are preserved after isolation. This method is then applied to study the influence of periodate treatment on nonspecific phagocytosis. In contrast with monocytes, we observed no decrease of phagocytic process. PMID- 6274441 TI - [Increased concentration of angiotensin I converting enzyme in the alveolar fluid of patients with sarcoidosis]. AB - Angiotensin converting enzyme (ACE) was assayed both in serum (SACE) and in bronchoalveolar fluid lavage (LACE) in 14 healthy controls and in 45 patients with sarcoidosis with mediastinal and pulmonary involvement. Concentration of SACE was 4466 +/- 2202 U x 100 ml-1 (mean +/- SD) in sarcoidosis and 2470 +/- 547 U x 100 ml-1 (chi +/- SD) in sarcoidosis and 2470 +/- 547 U . 100 ml-1 in controls. Concentrations of LACE were 65.2 +/- 48.4 U . 100 ml-1 and 21.1 +/- 14.7 U . 100 ml-1 respectively in sarcoidosis and in controls. These results are in favor of an intraalveolar secretion of ACE in sarcoidosis. LACE could be a better criterium than SACE for the evaluation of the pulmonary activity of sarcoidosis. PMID- 6274442 TI - Hepatic processing of IgA: studies using analytical and preparative subcellular fractionation. PMID- 6274443 TI - [Dynamics of the concentration of cyclic adenosine monophosphate in the tissues of rats with insulin hypoglycemia]. AB - The paper is concerned with the data on the cAMP content in rat tissues 15, 30, 60 and 120 minutes after insulin injection. Rapidly developing heterogenous changes were recorded in the cAMP content of rat tissues during insulin-induced hypoglycemia and stress reaction. The evidence obtained indicates that during stress reaction the line of the changes in the entire body depends on the triggering of numerous neurohumoral mechanisms that regulate the most essential processes of body activity via the adenylate cyclase system. PMID- 6274444 TI - [Study of mitochondrial permeability for labeled cAMP]. AB - Incubation of rat liver mitochondria with 3H-and 14C-cAMP under the conditions which are conducive to the activation of mitochondrial respiration by cAMP resulted in the penetration of cAMP into the mitochondria. Subfractionation of mitochondria was made with the use of digitonin and 0.1% Triton X-100. cAMP was incorporated into all the fractions investigated. A close correlation (r = +0.97) between 3H- and 14C-labelling of the mitochondria and submitochondrial fractions was found thus showing that the label was not only diluted but also accumulated by the mitochondria. PMID- 6274445 TI - [Sodium 2,3-dithiopropanesulfate blockade of the effect of cholera enterotoxin on adenylate cyclase and the concentration of cyclic 3',5'-adenosine monophosphate in the small intestine mucosa of the rabbit]. AB - Activation of adenylate cyclase in membrane preparations of the rabbit small intestine mucosa by cholera enterotoxin in the presence of sodium 2,3 dithiopropanesulfate (DTPS) is similar to that in the presence of dithiothreitol (DTT). Both DTT and DTPS do not influence the activity of adenylate cyclase without cholera enterotoxin. DTPS activates cyclic 3,5-AMP phosphodiesterase of mucosa homogenates (K0.5 = 10(-5) M). Combined administration of cholera enterotoxin and DTPS in situ into isolated loops of the rabbit small intestine decreases the activating effect of cholera enterotoxin on adenylate cyclase and diminishes the content of cyclic AMP in the mucosa. The destruction of disulfide bonds of cholera enterotoxin by DTPS is assumed to lower its ability to penetrate the mucosal cells of the small intestine. PMID- 6274446 TI - [Effect of various chemotherapeutic agents on DNA synthesis in the cells of human strains of breast and laryngeal cancer, chorionepithelioma and Wilms' tumor]. PMID- 6274447 TI - [Effect of cytostatics on the development of denervation disorders in skeletal muscles]. AB - Experiments on rats were made to study the effect of cytostatics on the rest membrane potentials (RMP) of muscle fibres and chemosensitivity of the botulinum toxin (BT) poisoned m. soleus. Intramuscular injection of the sublethal dose of BT on the 5th day evoked the blockade of the synaptic neuromuscular transmission, depolarization of the muscle cells and the decreased sensitivity to acetylcholine. Daily intraperitoneal injections of vincristine (25 micrograms/100 g) and fluorouracil (5 mg/100 g) to rats did not affect the development of the neuromuscular transmission blockade induced by BT. The cytostatics did not change the RMP of the myocytes or chemosensitivity of the normal muscles. However, both the drugs prevented the depolarization of myocytes and the decreased chemosensitivity of the muscles paralyzed with BT. It is assumed that the delayed appearance of the cytostatic-induced denervation is a consequence of the suppressed division of the satellite cells. PMID- 6274448 TI - [Effect of chronic administration of lithium chloride on the development of dopamine receptor sensitivity during morphine withdrawal in rats]. AB - The morphine withdrawal syndrome was studied in male Wistar rats. Spontaneous aggressiveness, enhanced apomorphine aggressiveness, lowered pain threshold and decreased dopamine turnover were observed after withdrawal of 10-day treatment with the increasing doses of morphine (30-300 mg/kg). These changes attested to the increased sensitivity of dopamine receptors. Administration of morphine in conjunction with lithium chloride in a dose of 2 mekv/kg prevented the development of dopamine receptor hypersensitivity. Also, this method did not produce the increased spontaneous and apomorphine aggressiveness or the decreased dopamine turnover. Meanwhile the pain threshold remained lowered. PMID- 6274449 TI - [Cytomorphological changes in human tumor cells exposed to high-polar compounds]. AB - The effects of some high-polar compounds on the cytomorphological characteristics of cultured cells of synovial and osteogenic human sarcomas (Sa-2 and Sa-4) were studied. Incubation of Sa-2 cells with 1 or 2% dimethylsulfoxide (DMSO) or 100 mM dimethylformamide (DMF) during 4-6 days induced the cellular, structural and functional changes in Sa-2 cells which were considered as manifestations of pseudoepithelial differentiation. DMSO did not influence the activity of Sa-2 cell enzymes, while DMF suppressed their activity, mainly that of acid and alkaline phosphatases. DMSO and DMF induced in Sa-4 cultured the appearance of the cells which were similar cytomorphologically to normal osteoblasts. DMSO and DMF significantly depressed the activity of acid phosphatase in Sa-4 cells and transformed the positive reaction of alkaline phosphatase to the negative one. DMSO and DMF prolonged the time of the doubling of Sa-2 and Sa-4 cells. The effects of DMSO and DMF were reversible. Methylformamide (200 mM) and dimethyl acetamide (0,1 and 10 mM) did not induce analogous changes in human sarcoma cells. PMID- 6274450 TI - [Effect of chemo- and radiotherapy on the growth of human strains of respiratory and excretory organ tumors transplanted in athymic mice]. AB - The effect of single and five-fold administration of 12 chemotherapeutic drugs on human strains of larynx, lung and kidney cancers and Wilms' tumor transplanted in nude mice was studied. Wilms' tumor was found to be more responsive to dactinomycin and cyclophosphamide. Larynx, lung and kidney cancers were resistant to all the chemotherapeutic drugs studied. The effect of irradiation of larynx cancer which resulted in short-term tumor growth retardation was also studied. PMID- 6274451 TI - [Repair processes in wound tissues of experimental animals following administration of glycine]. AB - A study was made of the effect of glycine given in doses approximating the physiological ones on the repair of processes in rat wound tissues. It was disclosed that in the early periods of wound healing, glycine administration leads to the increased content of cAMP and cAMP/cGMP ratio in the wound muscle and then in the granulation tissue, which appears to promote the intensification of the repair processes manifesting in the changes in tissue metabolism (DNA, collagen), in anti-inflammatory action, as well as in a more rapid maturation of the granulation tissue and wound reduction. It was also found that the doses of glycine tested do not affect the content of insulin and hydrocortisone in the blood of experimental animals. PMID- 6274452 TI - Human trophoblasts: cellular source of colony-stimulating activity in placental tissue. AB - Colony-stimulating activity (CSA) is a class of protein factors that stimulates the in vitro growth and differentiation of hematopoietic committed stem cells into mature granulocytes and macrophages. In man, production of CSA is mediated by monocytes-macrophages, lymphocyte, and endothelial cells. One of the best studied and most potent sources of CSA is conditioned media derived from cultured human placenta. The cellular source of this placental CSA was studied using cloned term placental cell lines induced by a simian virus 40 (SV-40) Wild-Type (wt) or temperature sensitive (tsA) mutants. Conditioned media from SV-40 wt transformed placental cells grown at 37 degrees C and tsA-transformed placental cells grown at 33 degrees C (the temperature at which the cells exhibit the transformed phenotype) and at 40 degrees C (temperature at which the cells express a normal differentiated phenotype) contained high levels of CSA. At the restricted temperature (40 degrees C), these cell lines expressed normal trophoblastic characteristics in that they produced human chorionic gonadotropin, pregnancy-specific-beta1-glyco protein, and placental alkaline phosphatase, indicating that the SV-40 transformation has not interfered with normal cellular functions of these trophoblasts. The CSA released by these cell lines resemble those of CM from fresh placental cells, in that the level of activity is high and that formation of both granulocyte/macrophage and eosinophil colonies is stimulated. PMID- 6274454 TI - Serum factors inhibiting E-rosette formation in patients with lung malignancies. PMID- 6274453 TI - The interaction of fibrinogen with human platelets in a plasma milieu. AB - Fibrinogen binds to specific receptors on human washed platelets and these sites are induced by adenosine diphosphate (ADP). This interaction is assumed to be the basis for the participation of the molecule in ADP-stimulated aggregation of platelets, but fibrinogen binding to platelets in plasma has not been directly demonstrated. In this study, we have characterized the interaction of 125I fibrinogen to platelets in the platelet-rich plasma (PRP) of afibrinogenemic patients. In either citrated or heparinized PRP, association of fibrinogen with platelets was demonstrable and was dependent on ADP dose. This binding reached equilibrium in 10-15 min, and saturation was achieved at fibrinogen concentrations greater than 0.5 microM. A linear Scatchard plot was derived that indicated a single class of binding sites with an affinity constant of Ka = 1.8 X 10(6) M(-1), and 32,000 fibrinogen molecules were maximally bound per platelet. The kinetics of the platelet fibrinogen interaction in plasma were essentially the same at 37 degrees C and 22 degrees C, but fewer molecules were bound at 37 degrees C. The rate constants of association were k 22 degrees C on = 0.9 X 10(6) M-1 . min-1 and k 37 degrees C on = 0.4 X 10(6) M-1 . min-1, respectively. Stabilization of the platelet-bound fibrinogen occurred only to a partial extent in both heparinized and citrated plasma. These results are similar to those obtained with washed platelets and establish that the previously defined steps in ADP-induced binding of fibrinogen to platelets occur in plasma, namely receptor induction by ADP, initial reversible binding, and irreversible binding. PMID- 6274455 TI - Cutaneous angiosarcoma arising in a mastectomy scar after therapeutic irradiation. AB - A cutaneous angiosarcoma developed in a mastectomy scar four years after post operative irradiation. Clinical and histological data are presented. The authors believe that radiotherapy was determinant in the genesis of this rare type of tumour. PMID- 6274456 TI - The uniqueness of protein sequences: a Monte Carlo analysis. PMID- 6274457 TI - Viruses and indoor air pollution. PMID- 6274458 TI - Inhalation of toxic products from fires. PMID- 6274459 TI - An autopsy case of Hodgkin's disease: intranuclear inclusion bodies in the tumor cells. PMID- 6274460 TI - Plasma cyclic-AMP and cyclic-GMP response levels in diabetes mellitus subjected to Cooper's aerobic programmed treadmill test. PMID- 6274461 TI - Insulin-induced enhancement of uptake of noradrenaline in atrial strips. AB - 1 Addition of insulin to the organ bath increased the force of contraction of guinea-pig left atrial strips driven electrically at 1 Hz. 2 The positive inotropic response to insulin remained unaltered in atria depleted of catecholamine or when beta-adrenoceptors were blocked by addition of propranolol to the organ bath. 3 The response os isolated atria to noradrenaline was significantly reduced in the presence of insulin. 4 Insulin affected neither the calcium accumulating abilities of the heart sarcolemma, mitochondria or microsomes, nor the cyclic adenosine 3',5'-monophosphate (cyclic-AMP)-protein kinase-induced stimulation of microsomal calcium uptake. 5 Addition of insulin to the organ bath enhanced significantly the ability of the cardiac tissue to take up [3H]-noradrenaline as well as [3H]-metaraminol. The activities of monoamine oxidase and catechol-O-methyl transferase were not changed after addition of insulin to homogenates of the heart. 6 The ability of insulin to facilitate uptake of noradrenaline would be expected to cause a decrease in the amount of the amine reaching the receptors, thus leading to a diminished response to this amine. This may explain, at least in part, insulin-induced subsensitivity to noradrenaline. 7 This view is supported by the observation that after blockade of amine uptake by destruction of nerve terminals, insulin failed to reduce the positive inotropic response to noradrenaline. PMID- 6274462 TI - Comparative studies of (--)-, (+/-)-propranolol, atenolol, guanethidine, bretylium and tetracaine on adrenergic transmission. AB - 1 Effects of (--)-, (+/-)-, and (+)-propranolol, atenolol, guanethidine, bretylium and tetracaine were studied on relaxation responses of rabbit ileum and contractile responses of rabbit pulmonary artery and guinea-pig vas deferens to electrical nerve stimulation (2 to 50 Hz). 2 In the ileum, inhibition by tetracaine 3.3 x 10(-6) M occurred at high frequencies of stimulation, while bretylium 1.2 x 10(-4) M and guanethidine 2 x 10(-5) M inhibited responses at all frequencies, the latter producing greater inhibition at low frequencies. 3 (+/-) Propranolol 10(-5) M produced a tetracaine-type inhibition after 1 h and a bretylium-pattern after 2 h in the ilea and pulmonary arteries and a transition from bretylium- to guanethidine-pattern in the vas deferens, while atenolol 2 x 10(-5) to 10(-4) M produced guanethidine-type inhibition in all preparations. 4 ( -)-, (+/-)-, and (+)-Propranolol 3 x 10(-6) to 3.3 x 10(-5) M were equipotent in the vas deferens and ileum. However, inhibition by (--)-propranolol 3.3 x 10(-5) M persisted in the ileum, while that by the (+)-isomer was partially restored by washing. 5 (--)- or (+)-Propranolol 3.3 x 10(-5) M or atenolol 2 x 10(-5) M did not inhibit relaxation of the ileum after the bath temperature was maintained at 4 degrees C for 2 h during drug application. 6 In conclusion, propranolol and atenolol both have gradually developing guanethidine-like adrenergic neurone blocking actions. PMID- 6274463 TI - Dissimilar effects on body temperature in the cat produced by guanosine 3',5' monophosphate, acetylcholine and bacterial endotoxin. AB - 1 Guanosine 3',5'-monophosphate (cyclic GMP) and N2-2'-O-dibutyryl guanosine 3',5'-monophosphate (db cyclic GMP) have been injected into the third cerebral ventricle (i.c.v.) of the unanaesthetized cat and the effects of rectal temperature and on behavioural and autonomic activities observed and compared with those of acetylcholine and physostigmine. 2 Acetylcholine (100 nmol) and physostigmine (100 nmol) injected together i.c.v. produced a rise in body temperature in cats at an environmental temperature of 20-24 degrees C, which was abolished by pretreatment i.c.v. with atropine (200 nmol). 3 Cyclic GMP and db cyclic GMP (10--1250 nmol) had no effect on body temperature in cats at an environmental temperature of 20--24 degrees C but produced hypothermia (1250 nmol) in cats at an environmental temperature of 9--11 degrees C. 4 The O-somatic antigen of Shigella dysenteriae (20 microgram/kg i.v.) produced fever in cats which was not potentiated by caffeine (25 mg/kg i.p.). Levels of endogenous cyclic GMP in c.s.f. taken from the cisterna magna during fever induced by bacterial endotoxin in the presence or absence of paracetamol (50 mg/kg i.p.) and/or caffeine were similar to values for afebrile cats. 5 It is concluded that exogenous cyclic GMP and db cyclic GMP can inhibit central events mediating autonomic and behavioural thermoregulation stimulated in cats by exposure to cold environments. PMID- 6274464 TI - Clonidine activates membrane potassium conductance in myenteric neurones. AB - 1 Intracellular recordings were made from neurones in the myenteric plexus of the ileum removed from guinea-pigs. The effects of clonidine and adrenaline on membrane potential and resistance were observed.2 Clonidine (100 pM-30 nM) caused a concentration-dependent membrane hyperpolarization associated with a fall in neurone input resistance.3 The amplitude of the clonidine hyperpolarization, but not the conductance increase, was greater in cells with lower resting potentials and smaller in more polarized neurones. In a given cell, membrane hyperpolarization decreased and membrane depolarization increased the clonidine effect.4 Low potassium solutions enhanced and high potassium solutions reduced the hyperpolarizing action of clonidine but did not significantly change the conductance increase caused by clonidine.5 The concentration-effect curve for clonidine was displaced to the left when the extracellular calcium concentration was reduced. Conversely, clonidine was almost ineffective in elevated calcium concentrations. This was true for both the hyperpolarization and the conductance increase.6 It is suggested that clonidine activates a potassium conductance by causing an elevation in the free intracellular calcium concentration.7 Clonidine reversibly depressed the amplitude of the nicotinic fast excitatory postsynaptic potential and the noncholinergic slow excitatory postsynaptic potential.8 All the effects of clonidine were shared by adrenaline and the actions of both were reversed or prevented by phentolamine (100 nM-1 muM). PMID- 6274465 TI - The inflammatory effects of hydroperoxy and hydroxy acid products of arachidonate lipoxygenase in rabbit skin. AB - 1. The inflammatory effects of hydroperoxy (HPETE) and hydroxy (HETE) acids, synthesized by arachidonic acid lipoxygenases, have been investigated in rabbit skin. 2. High doses (10-20 micrograms) of 5-, 12- or 15-HPETE or the 5,12-di hydroxy acid, leukotriene B4 (0.1-1 micrograms), caused small but significant increases in plasma exudation following intra-dermal injection. 3. Leukotriene B4 was equipotent with prostaglandin E2 and prostacyclin in potentiating bradykinin induced plasma exudation, and was 100 times more active in this property than any other lipoxygenase product tested. 4. Leukotriene B4-induced plasma exudation was enhanced by prostaglandin E2. 5. The mono-HETEs were relatively inactive in causing or enhancing plasma exudation. 6. Leukotriene B4 (0.1 microgram) or prostaglandin E1 (1.0 micrograms) significantly elevated leukocyte accumulation in rabbit skin, whereas PGE2, 5-HPETE, 5-HETE, 12-HPETE or 12-HETE were inactive at doses up to 1 microgram. PMID- 6274467 TI - Radioprotection of mouse skin by WR-2721 in single and fractionated treatments. AB - The radioprotective action of WR-2721 on mouse skin has been studied using single doses, two and five fractions. Significant radioprotection was observed with 200 500 mg/kg, in animals breathing air or oxygen at the time of irradiation. The degree of radioprotection increased with increasing dose, and was significantly greater in air than in oxygen, especially at low drug doses. Cumulative drug toxicity limited the fractionated treatments to lower drug doses, but the protection observed was similar to that seen with single doses at the same drug levels. The protection factors observed are lower than many published values for skin, and we conclude that a single protection factor cannot yet be stated for any dose of WR-2721. Local tissue oxygenation may account for some of the discrepancies in different studies. PMID- 6274466 TI - Electrical properties of smooth muscle cell membrane and neuromuscular transmission in the guinea-pig basilar artery. AB - 1 The membrane properties of smooth muscle cells and neuromuscular transmission in the guinea-pig basilar artery were investigated by use of microelectrodes.2 The membrane potential was -47.0 mV and the muscle tissue possessed cable-like properties as determined by the current-voltage relationships. The mean value of the space constant was 0.78 mm.3 An outward current produced a graded response and, in some cases, spike generation. This membrane response was enhanced in the presence of tetraethylammonium (TEA, 5 mM), and an increased concentration of TEA (10 mM) generated spontaneous spikes in most of the cells. Action potentials induced by TEA were abolished in the presence of MnCl(2) (5 mM) but not by isoprenaline (4 x 10(-6) M).4 Acetylcholine (ACh), over 10(-7) M, hyperpolarized the membrane and decreased the membrane resistance. This hyperpolarization increased in the presence of low [K](o) (below 5.9 mM), but decreased in [K](o) concentrations over 17.8 mM. Pretreatment with atropine (10(-6) M) suppressed the ACh-induced hyperpolarization. Therefore, this action of ACh is due to an increase in the K-conductance of the membrane produced by activation of the muscarinic receptors.5 Noradrenaline in concentrations up to 10(-4) M did not modify the membrane potential and resistance, while 10(-5) M, histamine, 5 hydroxytryptamine and adenosine triphosphate (ATP) depolarized the membrane. The depolarization induced by histamine or ATP was suppressed by reducing [Na](o). The histamine-induced depolarization was accompanied by an increase and the ATP induced one by a decrease in the membrane resistance. The action of histamine was suppressed by treatment with H(1)- but not H(2)-receptor blocking agents (dephenhydramine and cimetidine, respectively).6 Perivascular nerve stimulation (0.2 ms pulse duration) evoked excitatory junction potentials (e.j.ps). An increase in the number and frequency of stimuli enhanced the e.j.p. amplitude. In the presence of 1 mM TEA, a spike was evoked on the e.j.ps. A very high concentration of phentolamine (3.6 x 10(-4) M) or the usual concentration of tetrodotoxin (10(-7) M) abolished the generation of e.j.ps. Spontaneously generated miniature e.j.ps were never recorded from the resting membrane.7 The results are discussed in relation to regional specificities of smooth muscle cells of cerebral arteries in the guinea-pig. PMID- 6274468 TI - 99mTechnetium DMSA and the prediction of recovery in obstructive uropathy. PMID- 6274469 TI - Features of mammorgraphically negative breast tumours. AB - Mammography is a highly sensitive and specific technique in the diagnosis of breast cancer, but some tumours show no radiological features of malignancy. A total of 9.3 per cent of 323 consecutive patients with operable breast cancer had negative mammographic reports, and the clinical radiological and pathological findings in these cases are compared with those in the mammographically diagnosed group. Misinterpretation of carcinoma as a benign lesion and diagnostic difficulties in the dense dysplastic breast were found to be the main causes of error. PMID- 6274470 TI - The treatment of pleomorphic adenomas by formal parotidectomy. AB - We reviewed 100 patients who underwent formal parotidectomy. Seventy-two were new patients in whom a parotid lump proved to be a pleomorphic adenoma. Twenty-eight were patients in whom a tumour had recurred after treatment for pleomorphic adenoma by local excision (with or without irradiation) elsewhere. After a follow up of 1--15 years (median 4 years), there were no recurrences in the primary group but 5 recurrences among the secondary cases. Permanent facial nerve damage was not seen in the primary group, but 11 patients undergoing secondary parotidectomy were left with some permanent facial weakness. Malignant change occurred in 4 patients in the secondary group. PMID- 6274471 TI - A double-blind trial of suloctidil v. placebo in intermittent claudication. AB - In a recent double-blind trial lasting over 6 months, 40 patients suffering from intermittent claudication were randomly allocated to receive 300 mg of suloctidil per day or exactly matching placebo capsules. In addition to treadmill walking distance, other objective criteria including ankle blood pressure response and muscle blood flow measured by 133Xe clearance were used to assess the effectiveness of therapy. Nine patients (4 in the suloctidil group and 5 controls) did not complete the trial according to the protocol. Of the remaining 31 patients, 17 were in the control group and 14 received suloctidil. A significant improvement in the absolute walking distance, the level of beta thromboglobulin (beta TG) compared to pre-therapy value and the time for the ankle pressure index to return to the pre-exercise value was observed in patients receiving suloctidil. PMID- 6274472 TI - Gene action and the analysis of behaviour. PMID- 6274473 TI - Evidence for a raised concentration of a circulating sodium transport inhibitor in essential hypertension. AB - A cytochemical technique that measures the ability of plasma to stimulate guinea pig renal glucose-6-phosphate dehydrogenase (G6PD) activity in vitro, which is a marker of its ability to inhibit Na+-K+-adenosine-triphosphatase (Na+-K+-ATPase), was used in 19 patients with essential hypertension and 23 normotensive, healthy subjects. The ability of plasma to stimulate G6PD was significantly greater in the hypertensive patients when they were taking their normal sodium diet than in the normotensive subjects, and was significantly correlated with blood pressure. The ability of plasma to stimulate G6PD was inversely correlated with plasma renin activity in the hypertensive patients and increased with age and sodium intake in the normotensive subjects. These results support the hypothesis that essential hypertension, and also perhaps the increase in blood pressure with age in communities that consume large quantities of salt, is in part due to an increase in a circulating concentration of an inhibitor of Na+-N+-ATPase. PMID- 6274474 TI - Human papovavirus isolated from urine of a child with acute tonsillitis. PMID- 6274475 TI - Outbreak of rotavirus gastroenteritis among premature infants. PMID- 6274476 TI - Experience with selective venous sampling in diagnosis of ACTH-dependent Cushing's syndrome. AB - Twenty-three patients with adrenocorticotrophic hormone-(ACTH)-dependent Cushing's syndrome were subjected to selective venous catheterisation and sampling for ACTH on a total of 26 occasions. Out of 10 patients with pituitary dependent disease, nine had raised ACTH concentrations in one or both high internal jugular vein samples. Eight patients had 11 proved sites of ectopic hormone production: of these, six were correctly identified by the sampling technique, and in four of them this was the only accurate method of localisation. The results of one catheterisation were misleading, and on 10 occasions they were inconclusive; five patients remained undiagnosed by any method. Overall, 15 of the 26 catheterisations provided diagnostically valuable information. Selective venous catheterisation and sampling for ACTH is effective in confirming a pituitary source of the hormone and may be valuable in locating the source of ectopic ACTH production in some cases. PMID- 6274477 TI - The noradrenergic system in cultured aggregates of fetal rat brain cells: morphology of the aggregates and pharmacological indices of noradrenergic neurons. AB - Spherical aggregates formed rapidly in culture by re-aggregation of trypsin dissociated brain cells from the 17-day-old fetal rat. Over about 10 days in initially random distribution of cells evolved into a 3-layered arrangement; cells with characteristics of neurons were found largely in the intermediate layer. The survival of neuronal and glial cell types was evaluated histologically and verified by electron microscopy, which revealed synaptic and myelin structures that rapidly increased in number after 18 days in culture. Levels of norepinephrine (NE) and dopamine (DA) reached peaks of 9.5 and 4.4 ng/mg protein, respectively, at culture day 21. Uptake of [3H]NE paralleled these amine levels and was blocked by desipramine or pretreatment with either reserpine or 6-OH-DA. Autoradiographs of aggregates labeled with [3H]NE showed a high density of silver grains over cells, apparently neurons, with branching processes traced for 120 micrometer. Previously accumulated [3H]NE was released under depolarizing conditions (high [K+] or vertridine) only in the presence of Ca2+. Release was induced to a lesser extent by kainic greater than glutamic acid. Thus, such aggregates appear to contain catecholaminergic neurons capable of synthesis, uptake and release of NE. The time course of development of these functions supports suggestions that aggregate preparations might be useful in studying neurochemical or morphological aspects of brain development and function in vitro. PMID- 6274478 TI - Modification of neuronal activity in olfactory cortex slices by extracellular K+. PMID- 6274479 TI - Changes in prolactin binding sites in the rabbit hypothalamus induced by physiological and pharmacological variations of prolactin serum levels. PMID- 6274480 TI - Spiroperidol, naloxone, diazepam and QNB binding in the monkey cerebral cortex. PMID- 6274481 TI - [The use of silica as an anti-caking agent in food powder flavors]. PMID- 6274482 TI - [The use of silica in vegetable and powder soups]. PMID- 6274483 TI - [Beta adrenergic receptors on the branchial epithelial cells of trout (Salmo gairdneri) adapted to seawater]. PMID- 6274484 TI - [Adenosine-3',5'-phosphate, adenylate kinase, and cyclic AMP phosphodiesterase of the eye lens during aging]. AB - Compared to other tissues, the level of cyclic adenosine 3',5' monophosphate (cAMP) of the whole eye lens, at all ages, is rather low. In the epithelial cell layer cAMP content is of the same order of magnitude as in brain tissue but decreases considerably with age. Adenylate cyclase activities measurable only in the epithelial cells diminishes with age. Reduction of cAMP phosphodiesterase activity is observed during aging in lens fibre cells. PMID- 6274485 TI - Serum levels of 25-hydroxyvitamin D in postmenopausal osteoporosis. AB - Serum concentrations of 25-hydroxy-vitamin D were measured in a group of women with symptomatic postmenopausal osteoporosis, assessed by bone biopsy. A competitive protein binding assay was used, which included a chromatographic step. Accurate surveys of dietary or therapeutic vitamin D intake and light environment were obtained in each patient. Women with severe postmenopausal osteoporosis were found to have significantly (P less than 0.001) higher serum levels of 25-hydroxyvitamin D than age-matched normal women, the mean values being 27.5 ng/ml (+/- 13.6 SD) and 9.2 ng/ml (+/-5.7), respectively. The authors hypothesize that the reduction in 1,25-dihydroxyvitamin D, recently reported in postmenopausal osteoporotic women, might be responsible for the increased serum levels of 25-hydroxyvitamin D through an inadequate product inhibition of liver vitamin D 25-hydroxylase. PMID- 6274486 TI - Studies on the role of 24-hydroxylation of vitamin D in the mineralization of cartilage and bone of vitamin D-deficient rats. PMID- 6274488 TI - Effects of crude oil ingestion on avian intestinal function. AB - Intestinal function in mallard ducklings (Anas platyrhynchos) on a freshwater regime was studied after a 7-day dietary ingestion of 0.25% and 2.5% Prudhoe Bay crude oil (PBCO) or a 2.5% paraffin mixture with an in vivo luminal perfusion technique. Dietary ingestion of 2.5% PBCO may have an effect on the integrity of the duckling intestine. There were no significant effects of PBCO on the absorption of Na, Cl, K, or H2O compared with control animals. The ducklings fed 2.5% paraffin had a significant depression in Na and H2O absorption compared with controls. However, this depression dose not seem to be related to an effect on intestinal mucosa Na-K-ATPase activity. PMID- 6274487 TI - Biochemical and histological studies on various bone cell preparations. AB - Four different cell populations--designated PF, OB, OC, and PC--were isolated from calvaria of 18-day-old chick embryos for analysis of the effects of hormones on bone tissue. The cell populations were studied with histological and biochemical methods. Apart from the well-known cell types present in calvaria, a new cell type was found in the noncalcified organic matrix between the osteoblastic layer and calcified matrix. These cells were provisionally called osteocytic osteoblasts. They represent the "transition state" between osteoblasts and osteocytes. On the basis of histological studies with light microscopy (LM), transmission electron microscopy (TEM) and scanning electron microscopy (SEM), the PF population was considered to originate primarily from the periosteal fibroblasts, the OB population from the osteoblasts and osteocytic osteoblasts. The population of cells still present in calvaria from removal of periosteal fibroblasts and osteoblasts was called the OC population. This cell population was very much enriched with osteocytes. The fourth isolated population (PC) was a mixed population of fibroblasts, osteoblasts, and preosteoblasts. On exposure to parathyroid hormone (PTH), all four cell populations showed increased lactate production, but only the OB and OC populations displayed increased cAMP production. Prostaglandin E1 (PGE1) stimulated cAMP production in both OB and PF cells. From the results of this study it was concluded that PTH receptors are present on all of the cell types studied, but that occupancy of the receptor induces adenylate cyclase stimulation only in osteocytes and fully differentiated osteoblasts. PMID- 6274489 TI - Influence of vitamin D3 status, phenobarbital, and diphenylhydantoin treatment on the plasma 25-hydroxyvitamin D3 concentrations in the rat. PMID- 6274490 TI - Dibutyryl cyclic GMP, a competitive inhibitor of cholecystokinin/pancreozymin and related peptides in the gallbladder and ileum. PMID- 6274491 TI - Effect of digitonin on rat myometrium subcellular membrane fractions. AB - Isopycnic centrifugation experiments using sucrose density gradients showed that in digitonin-treated microsomes the distribution of the plasma membrane (PM) marker 5'-nucleotidase was shifted to higher densities. The treatment also caused similar but less pronounced changes in the distribution of protein, the putative endoplasmic reticulum (ER) marker NADPH-dependent cytochrome c reductase, and the inner mitochondrial marker cytochrome c oxidase. Similar experiments using more purified membrane fractions showed that the digitonin treatment led to a comparable increase in the densities of the fractions N1 and N2 previously described as subfractions of plasma membrane and to considerably less increase in the density of the fraction N3B which is enriched in the endoplasmic reticulum and the inner mitochondrial markers. Digitonin inhibited the ATP-dependent Ca uptake by the N1 fraction in a concentration-dependent manner (I50 = 0.3 mg/mL). Digitonin (0.5 mg/mL) inhibited the ATP-dependent azide-insensitive Ca uptake by all the fractions. The results support the hypothesis that (a) N1 and N2 are subfractions of plasma membrane, and (b) ATP-dependent azide-insensitive Ca uptake in rat myometrium is a property of plasma membranes. PMID- 6274492 TI - Thallous ion activation of p-nitrophenylphosphatase of rat myometrium plasma membrane. AB - Addition of thallous ion (Tl+) inhibited the spontaneous mechanical activity of rat myometrium in K-free Krebs solutions with an I50 value of 30 microM. The corresponding value for I50 for similar inhibition by addition of K+ was 150 microM. Tl+ as well as K+-activated p-nitrophenylphosphatase (PNPPase) of isolated rat myometrium plasma membrane vesicles. Half maximal activation was caused by 0.47 mM Tl+ or 1.6 mM K+. Maximal enzyme activities obtained using Tl+ and K+ were comparable. The Km values for the substance p-nitrophenylphosphatase using Tl+ (1.24 mM) and K+ (1.46 mM) were also similar. Activation by either ion was inhibited by ouabain, Na+, inorganic phosphate, and vanadate (V +5). The results suggest that Tl+ can substitute for K+ for activation of the Na-K pump of rat myometrium plasma membrane. PMID- 6274493 TI - The contribution of sensory inputs to the pattern generation of breathing. AB - Current concepts of the basic neural control system and its modulation by afferent inputs are reviewed. It is emphasized that, in analogy with locomotion, the central pattern generator (CPG) for automatic metabolic respiration does not depend on any afferent feedback from receptors sensitive to the movements of the "pump," or the streams of pumped air, for its production of a rhythmic motor output provides the CPG receives some "drive" inputs above threshold and adequate bias. The operation for a variety of reflexes and feedback loops is of fundamental importance, however, for adapting the breathing pattern to the varying requirements for gas exchange to the many behavioural, nonmetabolic demands on the breathing apparatus which are competing with its primary metabolic control functions. The presentation is focussed also on available evidence that the respiratory CPG exerts powerful modulations on the transmission in these reflex pathways controlling the pattern of breathing and adjusting it to the various metabolic and behavioural demands. Mechanisms for "gating," "phasic gain changes," and "phase-dependent reflex reversal" are exemplified. PMID- 6274494 TI - Some properties of the human erythrocyte receptors for Neisseria gonorrhoeae. AB - The nature of the receptors for T1 and T4 Neisseria gonorrhoeae on erythrocytes and other cells was investigated. In general, cells of nonprimate origin contained few receptors for gonococci. Receptors for T4 gonococci were only uncovered when host cells were pretreated with trypsin. Trypsinization, while unnecessary for T1 adherence to erythrocytes, enhanced attachment in inverse proportion to original erythrocyte sensitivity. Receptors for T1 and T4 organisms on trypsinized and trypsin-neuraminidase-treated erythrocytes were blocked by concanavalin A and peanut lectins, respectively, but a distinction could be made between them with wheat germ lectin and galactose oxidase. Of a number of sugars tested as inhibitors, only D-galactose blocked adherence of T4 but was without effect on T1. While the identity of erythrocyte receptors is uncertain, likely candidates are "band 3" protein and glycophorin, by virtue of their galactose content, lectin binding capacity, and partial exposure on the outer surface of the erythrocyte. PMID- 6274495 TI - The electron transport chain of Bacterionema matruchotii. AB - The membrane fraction of Bacterionema matruchotii contains an electron transport chain with oxidizing activity for NADH and succinate. Respiration was inhibited by KCN, 2-heptyl-4-hydroxyquinoline-N-oxide, UV light irradiation and CO. UV light irradiation, analysis of membrane extracts, and reconstitution of respiration in UV light treated membranes suggested that respiration is mediated by a menaquinone derivative. The membranes contained cytochromes a, b, and c. Inhibition studies and the effect of KCN and CO on the cytochrome spectrum indicated the presence of an a+a3 cytochrome oxidase and cytochrome o. The membrane fraction from cells grown under O2-limiting conditions contained nitrate reductase activity. In B. matruchotii, electron transport is coupled to oxidative phosphorylation as judged by the effects of substrates and inhibitors on the intracellular ATP concentration. PMID- 6274496 TI - Inability to rescue viral genes from human cells biochemically transformed by herpes simplex virus type 2 DNA. PMID- 6274497 TI - Solubilization of microsomal-associated phosphatidylserine synthase and phosphatidylinositol synthase from Saccharomyces cerevisiae. AB - Membrane-associated cytidine 5'-diphospho-1,2-diacyl-sn-glycerol (CDP diacylglycerol):L-serine O-phosphatidyltransferase (phosphatidylserine synthase, EC2.7.8.8.) and CDP-diacylglycerol:myo-inositol phosphatidyltransferase (phosphatidylinositol synthase, EC 2.7.8.11) were solubilized from the microsomal fraction of Saccharomyces cerevisiae. A variety of detergents were examined for their ability to release phosphatidylserine synthase and phosphatidylinositol synthase activities from the microsome fraction. Both enzymes were solubilized from the microsome fraction with Renex 690 in yield over 80% with increase to specific activity of 1.6-fold. Both solubilized enzymatic activities were dependent on manganese ions and Triton X-100 for maximum activity. The pH optimum for each reaction was 8.0. The apparent Km values for CDP-diacylglycerol and serine for the phosphatidylserine synthase reaction were 0.1 and 0.25 mM, respectively. The apparent Km values for CDP-diacylglycerol and inositol for the phosphatidylinositol synthase reaction were 70 microM and 0.1 mM, respectively. Thioreactive agents inhibited both enzymatic activities. Both solubilized enzymatic activities were thermally inactivated at temperatures above 30 degrees C. PMID- 6274498 TI - Modification of the poliovirus capsid by ultraviolet light. AB - Ultraviolet (UV) irradiation of type I poliovirus resulted in a modified (M) particle that had lost infectivity, lacked ability to adsorb to HeLa cells, lacked VP4, and reduced in S value. Additional irradiation resulted in the loss of VP2, further reduction in S value, and permeability of the capsid to RNAse, This particle (C) as well as M contain the genome. Acid pH (5.5-65) and sulfhydryl-reducing substances (dithiothreitol. reduced glutathione, and L cysteine) inhibited UV-induced modification of the capsid. UV irradiation at alkaline pH (7.5-8.5) resulted in more extensive modification of the capsid than irradiation at neutral pH. Ionic compounds were found to inhibit the modifying reaction. PMID- 6274499 TI - The management of brain metastases in germ cell tumors. AB - Twelve patients, with brain metastases from germ cell tumors were treated with combined whole brain irradiation and systemic chemotherapy. Two prognostic categories were identified. Patients with single brain metastases had a mean survival of 29.9 months, while patients with multiple brain lesions had a mean survival of 13.7 months. Four of the six patients with single brain lesions remain alive at 13, 14, 35 and 41 months following the clinical diagnosis of brain metastasis. The improved survival among patients with single lesions can be attributed to the sensitivity to chemotherapy of their tumors. Meningeal carcinomatosis, a previously unreported complication of metastatic germ cell tumor, developed in two patients who were initially treated for single brain metastasis. Both patients developed a clinical picture suggesting meningeal invasion by tumor and both had cytologic evidence in the cerebrospinal fluid of malignant cells. PMID- 6274500 TI - Gastrointestinal metastases from malignant tumors of the lung. AB - Autopsy data of 423 cases of primary tumor of the lung over a 36-year period were evaluated for the presence of gastrointestinal tract metastases. Fifty-eight cases (14%) were found and were analyzed for histologic nature of tumor, anatomic location, symptomatology and complications. The most common histologic type of lung tumor causing gastrointestinal tract metastasis was squamous cell (19 cases, 33%), followed by large cell (17 cases, 29%), and oat cell (11 cases, 19%). The esophagus was the most common site of involvement (33 cases). Fourteen of the 33 cases were involved by direct extension of the tumor. The middle third of the esophagus had metastases more commonly (16/33, 49%) than the other two sites. Most patients with gastrointestinal metastases had no symptoms. In those patients with symptoms, dysphagia was most common when the tumor involved the proximal gastrointestinal tract (esophagus, stomach), whereas, pain was most commonly seen with involvement of the distal gastrointestinal tract (small bowel, large bowel). Six of 20 patients (30%) with small bowel involvement experienced perforation and peritonitis as complications of metastatic involvement and two patients with large bowel metastasis had obstruction; a third had dehiscence of a previous anastomotic site. Gastrointestinal tract metastases from primary carcinoma of the lung are more common than previously thought and may be associated with serious clinical complications. PMID- 6274501 TI - Meningeal carcinomatosis 21 years following bronchiolo-alveolar carcinoma: diagnosis by cisternal CSF examination. AB - An unusual case is described of pure meningeal carcinomatosis in the absence of other evidence of tumor occurring 21 years after lobectomy for a localized bronchiolo-alveolar carcinoma. The metastatic disease was confined to the leptomeninges and was documented only by a cisternal cerebrospinal fluid (CSF) examination. Four previous lumbar punctures had shown hypoglycorrachia as the only CSF abnormality. PMID- 6274502 TI - Small cell carcinoma of the ovary with hypercalcemia: a report of eleven cases. AB - Eleven cases of a small cell ovarian cancer associated with hypercalcemia that was reversed by removal of the tumor are reported and compared with three similar cases in the literature. The 14 cases account for 50% of recorded cases of ovarian-cancer-related hypercalcemia. The 14 tumors occurred in women between the age of 13 and 35, with an average of 22 years and, with two exceptions, behaved clinically in an aggressive fashion. Light and electron microscopic study confirmed the epithelial nature of the neoplasms but failed to disclose their specific subtype or origin. PMID- 6274503 TI - Clinical hyperthermia: results of a phase I trial employing hyperthermia alone or in combination with external beam or interstitial radiotherapy. AB - Forty-three patients with advanced, locally accessible neoplasms were treated in a Phase I clinical trial employing hyperthermia alone or hyperthermia combined with either high-dose-rate external beam or low-dose-rate interstitial radiotherapy (interstitial thermoradiography). All patients had failed previous conventional therapeutic attempts, including various combinations of surgery, chemotherapy and radiation therapy. Many had received tolerance or near tolerance levels of prior radiation that restricted dose prescriptions in this trial to subcurative values. A number of tumors with different histologies were treated, including squamous cell carcinoma (14), adenocarcinoma (14), melanoma (8), malignant fibrous histiocytoma (2), and sarcoma (5). The response evaluation criteria used included no response (NR--less than 50% decrease in tumor volume), partial response (PR--50% less than or equal to tumor volume reduction less than 100%) and complete response (CR--complete tumor disappearance). For all tumor types, hyperthermia therapy alone resulted in a total response rate of 45% (27% PR, 18% CR). Hyperthermia combined with high-dose-rate external beam radiotherapy yielded a total response rate of 80% (53% PR, 27% CR). Seventeen patients treated with interstitial thermoradiography displayed a 100% total response rate (29% PR, 71% CR). By tumor histologies for all treatment groups, total response rates have ranged from 50% to 79% for all types except melanoma, which has shown a 100% (8/8) response rate to date. Response durations have varied from one to 24 months. Twelve of the 43 patients remain alive; three have no evidence of disease (NED) while nine have either stable local disease or are NED in the treated volumes but have metastatic disease. Complications have been minimal and have included one third-degree burn and three second-degree burns from fringing RF fields, one vaginal-rectal fistula, a superficial focal soft tissue necrosis, and some minor blistering. The results of this Phase I trial demonstrate that hyperthermia alone or combined with radiation can be safely applied in the treatment of malignant disease. Most importantly, the data suggest that hyperthermia, especially when combined with interstitial thermoradiography, can yield remarkable results in the eradication of local cancers. PMID- 6274504 TI - Erythroleukemia and other hematologic complications of intensive therapy in long term survivors of small cell lung cancer. AB - Eight patients with small cell bronchogenic carcinoma treated with intensive combination chemotherapy, with and without radiotherapy, have been followed for a minimum of two and a half years without relapse. One patient, after a prodrome of macrocytic sideroblastic anemia, leukopenia, and thrombocytopenia, experienced erythroleukemia 34 months after starting chemotherapy, and cytogenetic studies revealed extensive chromosomal abnormalities. Another patient had persistent macrocytic anemia and pancytopenia two years after cessation of therapy. The remaining six patients had normal peripheral blood smears and cell counts. A significant incidence of preleukemia syndromes and acute leukemia appearing as late complications in intensively treated small cell lung cancer patients requires confirmation in larger series of long-term survivors. Prospective determination of marrow karyotype abnormalities may help to identify patients at greatest risk for developing secondary leukemia. PMID- 6274505 TI - Meningosis prophylaxis with intrathecal 198Au-colloid and methotrexate in childhood acute lymphocytic leukemia. AB - Since 1972, telecobalt irradiation plus intrathecal methotrexate (ITMTX) has been successfully replaced in Jena by intrathecal colloidal radioactive gold (198Au) plus ITMTX for meningosis prophylaxis in leukemia. Seventy-three children with acute lymphocytic leukemia (ALL) were given 1.24-4.89 mCi (45.8-181 MBq) of colloidal 198Au IT after successful initiation of remission. During cytostatic therapy, the following relapses occurred: meningosis leucaemica, five patients (6.8%); bone-marrow relapse and the meningosis leucaemica, one patient; and bone marrow relapse, 20 patients (27.4%). In 18 children, combination chemotherapy was terminated after two and a half or three years of treatment. After that time, one meningeal relapse and six bone-marrow relapses occurred. Within the first 24 hours after application of radioactive gold, headaches, vomiting, and fever occurred in less than 10% of the children. An apathy syndrome, leukecephalopathy, or severe infections, were not observed in a single case. Radioactive gold spreads in the subarachnoid space and is phagocytized by the arachnoidea. The tumoricide effect extends selectively over the space of distribution of the latent meningosis leucaemia. The cerebral parenchyma remains unaffected by radiation. Thus, radioactive gold may be preferable to telecobalt irradiation in preventing central nervous system leukemia. PMID- 6274507 TI - Sarcomatous transformation of nasopharyngeal angiofibroma. AB - A case of fibrosarcoma arising in a recurrent nasopharyngeal angiofibroma 18 years after radiation therapy is described. A review of the medical literature revealed two other documented cases of sarcomatous transformation of angiofibroma, and in both, the angiofibromas had also been irradiated before the sarcomatous transformation. These occurrences should caution against the indiscriminate application of radiation therapy in nasopharyngeal angiofibromas. PMID- 6274506 TI - Diagnosis of primary breast carcinoma through immunohistochemical detection of antigen related to mouse mammary tumor virus in metastatic lesions: a report of two cases. AB - Recent investigations have established that approximately half of human breast carcinomas contain an immunohistochemically detectable antigen which is cross reactive with the 52000-dalton major glycoprotein (gp52) of the mouse mammary tumor virus (MMTV). This antigen can be localized in paraffin-embedded sections of routinely fixed tissues using heterologous antibodies to gp52 or MMTV. This report describes two patients with metastatic carcinoma in axillary lymph nodes without any clinical evidence of a primary lesion in the breast or elsewhere. The localization of the gp52-related antigen in paraffin-embedded sections of both metastatic lesions suggested the presence of primary mammary carcinoma. In both instances, this suggestion was ultimately confirmed by the finding of primary lesions in which the gp52-related antigen was also found. PMID- 6274508 TI - Carcinoembryonic antigen in carcinoma of the uterine cervix: antigen distribution in primary and metastatic tumors. AB - Immunohistochemical staining for carcinoembryonic antigen (CEA) was performed on primary tumors and regional lymph nodes from 100 patients undergoing radical hysterectomies and pelvic lymphadenectomies for invasive carcinoma of the uterine cervix. Antigen staining was present in 82% of keratinizing squamous carcinomas as compared with 50% in nonkeratinizing tumors. Seventeen patients with CEA producing cervical cancer had regional lymph nodes metastases, all of which stained positively for CEA. Conversely, antigen could not be detected in lymph nodal metastases from primary tumors devoid of CEA. The pattern and intensity of CEA staining in primary tumors and their metastases were similar in all cases. Antigen was present in highest concentrations in the cytoplasm and tumor cell membrane of all tissues examined and isolated nuclear staining was absent. The clinical implications of these findings are discussed. PMID- 6274509 TI - Oral contraceptives and hepatocellular carcinoma. AB - A 23-year-old woman was seen with an acute hemoperitoneum caused by rupture of a hepatocellular carcinoma. She died two years after hepatic artery ligation. The patient had been taking oral contraceptives for periods of one year and ten months, separated by a pregnancy. The association between oral contraceptive steroids and hepatocellular carcinoma is briefly reviewed. PMID- 6274510 TI - Malignant lymphoproliferative diseases induced by Epstein-Barr Virus in immunodeficient patients, including X-linked, cytogenetic, and familial syndromes. PMID- 6274511 TI - Quantitation of differences between spontaneous and induced liver tumors in mice with an automated image analyzer. AB - Spontaneous hepatocellular neoplasms of B6C3F1 mice (basophilic neoplasms) as well as those induced with the herbicide Nitrofen (eosinophilic neoplasms) were observed with an image analyzing computer to determine if quantifiable morphologic differences existed between them. Several morphologic parameters were measured on 5 liver sections from each of the following groups: (a) unexposed control liver; (b) non-trabecular basophilic neoplasms; (c) trabecular basophilic neoplasms; (d) small well circumscribed eosinophilic neoplasms; and large irregular eosinophilic neoplasms without (e) and with (f) pulmonary metastases. The total number of hepatocytes per unit area was significantly smaller in the eosinophilic neoplasms than in the basophilic neoplasms or the controls. This was the result of a greatly increased cell cross-sectional area in the eosinophilic neoplasms, caused predominantly by a larger cytoplasmic cross-sectional area. Nuclear cytoplasmic ratios of cells in eosinophilic neoplasms were lower than in the other groups for this reason. This demonstrates that quantitative morphologic differences exist between the spontaneous and induced neoplasms, which supports the conclusion that Nitrofen is a true carcinogen, and not a promoting agent. PMID- 6274512 TI - Expression of human fetal brain antigens by human tumors of neuroectodermal origin as defined by monoclonal antibodies. AB - The antigenic relationships between human tumors of neuroectodermal origin and fetal brain were investigated by the production of hybridoma antibodies derived from a fusion of P3-NS1/1-Ag 4-1 (NS1) myeloma cells with splenocytes from a mouse multiply immunized with an homogenate of second-trimester human fetal brain tissue. Two monoclonal antibodies (MAs), 4D2cl 6 and 7H10cl 4, were studied in detail by cell surface radioimmunoassay (CS-RIA), quantitative absorption, indirect immunofluorescence, and peroxidase-anti-peroxidase (PAP) immunohistology. MA 4D2cl 6 binds to 5 of 14 glioblastoma (GBM) cell lines, 1 of 2 melanoma cell lines, 1 of 3 neuroblastoma cell lines, and 1 of 5 fetal fibroblast lines by CS-RIA and to 13 of 13 GBM, 1 neuroblastoma, and fetal brain, liver, spleen, and adult spleen unfixed frozen tissue by PAP analysis. MA 7H10cl 4 binds to 13 of 14 GBM, 1 of 3 neuroblastoma, and 1 medulloblastoma cell line(s) by CS-RIA analysis and to 13 of 13 GBM, 1 neuroblastoma, fetal brain, liver, spleen, thymus, and adult spleen by PAP analysis. Control non-central nervous system tumors and normal adult tissue, including brain, were unreactive with both MAs by CS-RIA, PAP, and absorption analysis. Tissue distribution and localization analyses established that MAs 4D2cl 6 and 7H10cl 4 recognize specificities of shared fetal-neuroectodermal-lymphoid distribution which are operationally specific within the adult central nervous system and which are not related to previously described oncofetal or onconeural antigens. PMID- 6274513 TI - Effects of the phorbol ester 4-O-methyl-12-O-tetradecanoylphorbol-13-acetate on mouse skin in vivo: evidence for its uselessness as a negative control compound in studies on the biological effects of phorbol ester tumor promoters. PMID- 6274514 TI - O-and C-D-glucosyluronic acid derivatives of delta 1-tetrahydrocannabinol: synthesis and differential behavior to beta-glucuronidase. PMID- 6274515 TI - The (Na+ + K+)-stimulated ATP-A from Yoshida sarcoma ascitic tumor cells- comparison of the properties of partially purified and highly purified enzyme. PMID- 6274516 TI - Quantitative topography of organelles in the liver. A combined histochemical and morphometric analysis. AB - After seven days of feeding fructose the liver of Wistar rats showed enormous accumulations of glycogen, which completely altered the original pattern of distribution of organelles. A quantitative morphological method was used to analyze these changes. The cytoplasm was mapped into arbitrary "distance classes" corresponding to concentric rings beginning at the outer nuclear membrane. This allowed the density of organelles in a given zone to be estimated. In cells filled with glycogen as a result of the fructose feeding, the following rearrangements were found: in the intermediate zone of both cellular poles (i.e., bile canalicular pole and sinusoidal pole) the mitochondria disappeared, being replaced by glycogen. The endoplasmic reticulum was accumulated in the perinuclear zone of both cellular poles, as in control animals, but was reduced throughout the rest of cytoplasm. It showed a peripheral density maximum at the biliary canalicular pole, in contrast to the cells of control animals. These changes in the distribution of the organelles and cellular "compartments" correspond to histochemical findings and demonstrate an adaptive reaction in the liver parenchyma to fructose ingestion, the organelles arranging themselves in cytoplasmic regions which still show a metabolic activity. PMID- 6274517 TI - Changes in corticotropic responsiveness and mitochondrial ultrastructure of adrenocortical cells from the inner zone of the duck (Anas platyrhynchos) adrenal gland: the effects of cycloheximide, puromycin, and chloramphenicol. PMID- 6274519 TI - Concomitant induction of cytotoxic and suppressor cells: modulation by theophylline. PMID- 6274518 TI - Calmodulin is involved in the first step of oocyte maturation: effects of the antipsychotic drug fluphenazine and of anticalmodulin antibodies on the progesterone-induced maturation of xenopus laevis oocyte. AB - Specific anticalmodulin antibodies were microinjected into full-grown Xenopus laevis oocyte, and it is shown that in ovo blockade of the complex Ca2+ calmodulin accelerates the kinetics of progesterone-induced maturation, even though the molar ratio of antibody binding sites to total calmodulin was only 0.16. Addition of 200 microM fluphenazine to the oocyte incubation medium resulted in a similar acceleration of steroid-induced maturation. Neither protein kinase inhibitor (PKI) nor maturation promoting factor (MPF)-induced maturation are affected either by the antipsychotic drug or by anticalmodulin antibodies; this result suggests that the adenylate cyclase system may be the target for anticalmodulin antibodies and fluphenazine effects. PMID- 6274520 TI - [Abrikosov's tumor (granular cell myoblastoma) with a laryngeal localization. Report of 2 cases]. PMID- 6274521 TI - Studies on antiviral glycosides: V. Formation of incomplete Sendai virions in the presence of phenyl-6-chloro-6-deoxy-beta-D-glucopyranoside. PMID- 6274522 TI - Facile desulfurization of thiocarbonyl groups to carbonyls by superoxide. A model of metabolic reactions. PMID- 6274523 TI - Potent orally active inhibitors of angiotensin-converting enzyme (ACE). PMID- 6274524 TI - [A preliminary study of hepatitis A virus in Chinese tupaia (author's transl)]. PMID- 6274525 TI - [Carriers of Salmonella among the people of four villages in Abidjan area (Ivory Coast) (author's transl)]. AB - The authors relate the results of a bacteriological investigation carried out among the people of four villages from Abidjan outskirts (about 1,704 inhabitants). They searched for individuals expelling Salmonellas from their faeces. This investigation, which lasted a year enabled, out of 1,754 cultures to isolate 109 strains belonging to various serotypes, four new ones among them. The isolations frequency increases with the rainy season, and has been especially important in one of the villages. Animals expell Salmonellas from their dejections, in a high percentage of cases. PMID- 6274526 TI - [Serological survey for the prevalence of certain arboviruses in the human population of the south-east area of Central African Republic (author's transl)]. AB - A serological survey of antibodies to arboviruses was carried out in the human population of the south-east part of Central African Republic in April 1979. Four hundred and fifty nine serum samples were tested using the haemagglutination inhibition test (H. I.) and fifty of them by the complement fixation test (C. F.). Only 11% of the population tested had no H. I. antibodies against the following arboviruses: Chikungunya, Semliki-forest, Sindbis, Yellow fever, Uganda S, West-Nile, Zika, Bunyamwera and Zinga. It seems that Chikungunya virus has recently been very active mainly in adult population. The same observation was reported for Zika virus. The following antigens were used for complement fixation test: Ilesha, Bwamba, CHF-Congo, Dugbe, Bhanja, Tataguine, Nyando, Bangui and Orungo. A positive reaction was noted in 88% of serum samples tested for Orungo virus. Antibodies were also detected by CF for CHF-Congo and Bhanja viruses. PMID- 6274527 TI - [Natural pathogenicity for man of an antigenic variant of Soldado virus from Morocco (author's transl)]. AB - Pathogenicity of Soldado virus was clearly established in the past for seabirds but remained questionable for man. In a scientist, repeatedly bitten by Ornithodoros (A.) maritimus larvae at Essaouira (Morocco), the association of fever of unknown origin and of neutralizing antibody against an antigenic variant of Soldado virus isolated from the same place, lead to the conclusion that this virus may act as a pathogen for man. Epidemiological implications of such an observation are briefly discussed. PMID- 6274528 TI - [Biological health check-up for woodworkers in Ivory Coast: first results (author's transl)]. AB - A random sample of 125 woodworkers in Ivory Coast was subjected to a biological health check-up. Being relatively simple this check-up has allowed us however to reveal the following pathological states for subjects not complaining of any illness: 2 anemia, 25 haemoglobinopathies, I monoclonal dysglobulinemia, I massive dysglobulinemia with splenomegaly, 8 massive hypergammaglobulinemia. Otherwise the following frequent values gave been determined according to RICHTERICH's statistical approach: age, height, weight, hematocrite haemoglobinemia, haemoglobin type, proteinemia, lipidemia, cholesterolemia, triglyceridemia, alpha-lipoproteinemia, A. M. G. immunoglobulinemia. PMID- 6274529 TI - [Hepatoma manifested by an orbito-sinus metastasis]. PMID- 6274530 TI - [Oculo-dento-osseous dysplasia]. PMID- 6274531 TI - Thoracic radiation therapy and Adriamycin/cisplatin-containing chemotherapy for locally advanced non-small-cell lung cancer. AB - One-hundred nine patients with limited-stage unresectable non-small-cell lung cancer were treated with Adriamycin--cisplatin-based combination chemotherapy and thoracic irradiation. Of this number, 73 received chemotherapy (one course) prior to irradiation and 37% (27/73) had tumor regression following chemotherapy alone. Sixty-eight percent of patients (73/107) experienced tumor regression following combined chemotherapy and irradiation. Age more than 65 years, a malignant ipsilateral pleural effusion, and no response to chemotherapy alone were all strong negative prognostic factors. Three-year survivals were as follows: all 109 patients, 14.0%; 89 patients without malignant pleural effusion, 17.0%; 71 patients with neither a malignant pleural effusion nor malignant ipsilateral supraclavicular adenopathy, 23.1%. PMID- 6274532 TI - Phase II trial of methylglyoxal-bis-(guanylhydrazone) in non-small-cell lung cancer. AB - Fifty-two patients with metastatic or recurrent non-small-cell lung cancer (NSCLC) were treated, during a phase II trial, with methylglyoxal-bis (guanylhydrazone) (MGBG). Of the 44 patients who had adequate trials, 4 had partial responses (PR), for an overall 9% PR rate. Response durations ranged from 3 to 5+ months. Prior treatment with chemotherapy may have adversely affected response rate; 15% of previously untreated patients responded, compared to only 4% of previously treated patients. A syndrome of weakness and fatigue was the most serious side effect. Anorexia and weight loss, stomatitis, nausea and vomiting, diarrhea, and peripheral neuropathy were the other toxic effects. We conclude that MGBG has activity in NSCLC, especially in previously untreated patients, and further studies are indicated in that population. PMID- 6274533 TI - A phase II trial of m-AMSA in patients with small-cell lung cancer. AB - Eighteen patients with small-cell lung cancer were treated with m-AMSA in doses of 90 mg/m2 or 120 mg/m2 intravenously every 3 weeks. There were no useful therapeutic responses in the 16 patients receiving adequate trials of the drug. Myelosuppression was the only significant dose-limiting side effect of treatment. m-AMSA in this dose and schedule is not sufficiently active in small-cell lung cancer to warrant further study in a heavily treated patient population. PMID- 6274534 TI - Phase I trials of WR2721 in combination with radiation therapy and with the alkylating agents cyclophosphamide and cis-platinum. AB - Three phase I trials of the radioprotector S-2-(3-aminopropylamino) ethylphosphorothioic acid (WR2721) have accessed 60 patients. Study 1 is devised to determine the maximum tolerated dose (MTD) of a single dose of the protector 15 to 30 minutes before a single radiation treatment of a size used routinely in palliative management. Study 2 plans to determine the MTD for up to 15 daily doses of the drug over 3 weeks during palliative radiotherapy. Also, the multipe dose study will establish the MTD before palliative irradiation for fewer than five fractions a week. Study 3 uses the existing single-dose MTD determined in Study 1 as pretreatment 15 to 30 minutes before cyclophosphamide or cis-platinum. Toxic symptoms include emesis, hypotension, hypertension, somnolence, and sneezing. Only one serious episode of hypotension, considered idiosyncratic, and one instance of moderate to severe vomiting have occurred. Forty-one patients have been entered in Study 1 and a dose of 600 mg/m2 has been reached. The next step is to proceed to the planned highest level of 740 mg/m2. Of five patients in the multiple-dose study, one has been given, without toxicity, WR2721 at the level of 100 mg/m2 for 15 fractions over 3 weeks. Fourteen patients are accessed to the alkylating agent study. Using protector doses of 450 mg/m2, a cyclophosphamide level of 1500 mg/m2 has been accomplished. However, two of three patients who received 450 mg/m2 of WR2721 before 120 mg/m2 of cis-platinum have shown moderate elevation of the serum creatinine, both of which returned to normal. PMID- 6274535 TI - A collaborative study of cytomegalovirus antibodies in mothers and young children in 19 countries. AB - Regional variations in the incidence of cytomegalovirus (CMV) injections in mothers and young children were investigated by testing cord blood and serum samples from infants and children up to four years old for the presence of CMV antibodies in 19 different regions of the world. The samples were tested by both the local virus laboratory and the reference laboratory, using the same batch of complement-fixing CMV antigen and techniques which were validated by comparison of the titres recorded for samples of coded sera sent to each laboratory. The incidence of CMV antibodies varied from 44-100% in mothers and from 3-95% in young children. The number of children with CMV antibodies increased with age in five areas; suggesting that there was some child to child transmission of CMV infection between children in these regions. In the other regions, the absence of any significant age-related increase indicated that the main pathway of CMV infection in early life was by transmission from mothers to their infants. The significance of these findings is discussed. PMID- 6274536 TI - Coliphages as ecological indicators of enteroviruses in various water systems. AB - The occurrence of coliphages and enteroviruses in a variety of water systems in Czechoslovakia was monitored for two years. Two host strains of Escherichia coli bacteria were used to test 1161 water samples for the presence of bacteriophages. These strains were polyvalent hosts for a broad spectrum of morphologically distinct coliphages, and their use thus gave quantitative data on the degree of viral pollution in any given water sample. Ninety-two water samples were tested in parallel for the presence of enteroviruses, by using a flocculation method to concentrate the viruses followed by isolation in cultures of a buffalo green monkey (BGM) kidney continuous cell line. The enterovirus and coliphage recovery rates showed similar differences when waters with different levels of pollution were compared. Seasonal fluctuations of both the coliphage and enterovirus (mostly poliovirus) levels in river water were demonstrated by statistical analysis of the data collected. The levels increased in the winter and sharply declined in the summer months as the river water temperature increased. Chemical pollution did not seem to influence the survival of either the coliphages or the enteroviruses in the observed rivers. PMID- 6274537 TI - Chemosensitivity of human neoplasms with in vitro clone formation. Experience at the University of Southern California - Los Angeles County Medical Center. AB - We analyze experience with 600 specimens for in vitro chemosensitivity assessment of human neoplasms utilizing a soft agar colony-forming technique. Good test reproducibility is demonstrated. Disaggregation with collagenase enhances yield and does not alter chemosensitivity profiles. Therapeutic exposure to chemotherapy prior to biopsy reduces in vitro sensitivity to the specific agents used in vitro. The cyclophosphamide derivatives 4-hydroperoxycyclo phosphamide (4 HC) and phosphoramide mustard are active in vitro, and produce comparable rank order sensitivities among tested tumors. There is marked reduction of in vitro 4 HC sensitivity in patients with prior therapeutic cyclophosphamide exposure, supporting the use of this derivative in test systems. Rank order of test results among specimens is compared at 0.1 microgram and 10 microgram drug/ml. Substantial differences in rank order at these two dose levels are demonstrated, indicating that the in vitro test dose selected is an important variable. PMID- 6274538 TI - Studies on the in vitro transfer of DNA binding benzo[a]pyrene metabolites from rat hepatocytes to human fibroblasts. AB - The release and reabsorption of benzo[a]pyrene (BP) metabolites were studied in isolated rat hepatocytes incubated with BP. There was a rapid uptake of BP by the hepatocytes which was followed by an excretion of organic as well as water soluble metabolites. Upon further incubation the BP-dihydrodiols and BP-phenols were reabsorbed by the cells and finally excreted as water-soluble conjugates. The metabolism of BP by the hepatocytes also resulted in the formation and release of reactive intermediates to the incubation medium. The DNA-binding intermediates released from the cells were consistent with trans-7,8-dihydroxy 7,8-dihydrobenzo[a]pyrene-9,10-oxide(s) and 9-hydroxybenzo[1]pyrene-4,5-oxide(s). The BP-metabolites were found to bind to hepatocyte and fibroblast DNA. The degree of DNA binding was estimated as the formation of DNA strand breaks. BP metabolites formed within the hepatocytes and transferred to the fibroblasts were found to induce DNA strand breaks in the latter cells. Addition of DNA to the medium markedly decreased the fraction of single stranded DNA in these cells. Under conditions where the excretion from hepatocytes of BP-phenols and BP dihydrodiols were increased, the amount of DNA strand breaks in the fibroblasts increased. PMID- 6274539 TI - Presence of benzo[a]pyrene metabolizing activities in isolated rat liver nucleoli. AB - A benzo[a]pyrene (B[a]P) hydroxylase activity and epoxide hydrolase (EH) activity have been found in rat liver nucleoli obtained from untreated (C) and 3 methylcholanthrene (3-MC) pretreated rats. A comparison of the enzyme activities was made in rat nuclei and nucleoli. Light and electron microscopic observations of nucleolar preparations did not reveal significant contamination either by intact nuclei or by nuclear membranes, and glucose 6-phosphatase, a marker of microsomal activity, was not detected in our nucleolar preparations. NADPH cytochrome c-reductase could be measured in C and 3-MC nuclei and very low but detectable activity was found in the nucleoli. Nucleolar preparations revealed significant hydroxylating activity, which was inducible by 3-MC in nuclei but not in nucleoli. The presence of EH in nucleoli was demonstrated with phenanthrene 9,10-oxide and B[a]P 4,5-oxide, but the nucleolar activities were lower than those obtained using intact nuclei. The addition of 1,1,1-trichloropropene 2,3 oxide completely inhibited EH activity. Furthermore the presence of covalently bound metabolites of B[a]P formed in isolated nucleoli was demonstrated by cytofluoromicroscopy. PMID- 6274540 TI - Inhibition of spontaneous hepatocarcinogenesis in C3H/HeN mice by Edi Pro A, an isolated soy protein. PMID- 6274541 TI - K+ fluctuations in the extracellular spaces of cardiac muscle. Evidence from the voltage clamp and extracellular K+ - selective microelectrodes. PMID- 6274542 TI - Mechanisms of action of lidocaine and quinidine on action potential duration in rabbit cardiac Purkinje fibers. An effect on steady state sodium currents? PMID- 6274543 TI - Studies of heterogeneity of angiotensin-converting enzyme and acid phosphatase in granulomatous lesions of skin. AB - Activities of angiotensin-converting enzyme and acid phosphatase were markedly elevated in the lesions of metal-induced hypersensitivity and foreign body types of granulomas. We biochemically studied these enzymes by means of gel filtration and electrophoresis and compared them with the same enzymes of normal dermis. Two forms of angiotensin-converting enzyme found in normal dermis were increased proportionally. Catalytic properties and heat stability of the enzyme in the lesions and normal dermis were the same. On the other hand, acid phosphatase in the lesions showed a different isozyme pattern from that seen in normal dermis. The enzyme from normal dermis had apparent molecular weight of 150000 and 90000, whereas an additional enzyme with a molecular weight of 230000 was detected in granulomatous lesions. Three isozymes were demonstrable in normal dermis but two additional isoenzymes appeared in granulomatous lesions: the isozyme patterns between the foreign body and hypersensitivity granulomas are different. PMID- 6274544 TI - The metabolic effects of chronic hyperglucagonaemia. AB - A subject with a benign glucagonoma was studied before and after complete resection of his pancreatic tumour. Studies were undertaken pre- and post operatively to determine the effects of chronic hyperglucagonaemia on glucose tolerance and glucose kinetics both in the fasting state and during physiological insulin infusions, employing the [3H]-3-glucose technique. In addition the plasma cyclic AMP response to an acute infusion of glucagon was studied pre- and post operatively. The basal immunoreactive glucagon levels pre- and post-operatively were 10492 +/- 1296 and 149 +/- 15 pg/ml respectively. Pre- and post-operative oral glucose tolerance tests did not differ but were abnormal. Pre-operatively basal hepatic glucose production was normal and it was suppressed rapidly by the low dose insulin infusion, despite continuing hyperglucagonaemia. The metabolic clearance rate of glucose was slightly reduced. There was no plasma cyclic AMP response to a glucagon infusion, suggesting down-regulation of the glucagon receptor by the chronic hyperglucagonaemia. Post-operatively the hepatic glucose production and clearance rate of glucose fell, whereas the plasma cyclic AMP responses to the glucagon infusion reverted to a normal pattern. It is concluded that chronic hyperglucagonaemia is not a major factor in the development of the glucose intolerance, but it may lead to down-regulation of the biological action of glucagon. PMID- 6274545 TI - Gamma-melanotrophin-like immunoreactivities in human pituitaries, ACTH-producing pituitary adenomas, and ectopic ACTH-producing tumours: evidence for an abnormality in glycosylation in ectopic ACTH-producing tumours. AB - Using gel exclusion chromatography on Bio-Gel P-60, gamma-melanotropin-like immunoreactivity (gamma-MSH-LI) in three human pituitary glands, two ACTH producing pituitary adenomas, and three ectopic ACTH-producing tumours (two medullary thyroid carcinomas and one thymoma) was divided into one or two molecular weight classes. The largest component eluted near the position of mouse 16K fragment and was designated big gamma-MSH-LI. This big gamma-MSH-LI was present in all samples. The second one, designated intermediate gamma-MSH-LI, eluted between the position of mouse 16K fragment and human ACTH, and was demonstrated only in two ectopic ACTH-producing tumours. No gamma-MSH-LI emerged at the elution position of synthetic gamma 3-MSH. Affinity chromatography on concanavalin A-agarose revealed that a significant fraction (52-68%) of gamma-MSH LI from human pituitary glands, ACTH-producing pituitary adenomas, and one ectopic ACTH-producing tumour bound to the column and was eluted with alpha methyl-D-mannopyranoside. In two ectopic ACTH-producing tumours which contained big and intermediate gamma-MSH-LI, a relatively small fraction (27-35%) of gamma MSH-LI bound to the column and was similarly eluted. These observations suggest that human gamma-MSH-LI is glycosylated and that there is an abnormality in the glycosylation of gamma-MSH-LI in some ectopic ACTH-producing tumours. PMID- 6274546 TI - Evidence for secondary hyperparathyroidism in the osteomalacia associated with chronic liver disease. AB - Previous reports have suggested that secondary hyperparathyroidism is extremely uncommon in hepatic osteomalacia. This, together with other findings, has led to suggestions that in chronic liver disease there may be selective resistance of bone to vitamin D or a specific bone mineralization defect unrelated to Vitamin D. To examine these possibilities, twenty-five patients with chronic liver disease have been studied by bone biopsy, serum calcium and inorganic phosphate, plasma 25-hydroxyvitamin D, plasma immunoreactive parathormone (iPTH), fasting urine cAMP, fasting renal tubular maximal reabsorptive capacity for phosphate (TmP/GFR) and fine grain hand x-rays. Nine of the patients had osteomalacia on bone biopsy, eight of these had subnormal levels of plasma 25-hydroxyvitamin D and the other had a borderline result. Based on the consensus of all the tests, five of these had evidence of secondary hyperparathyroidism. Plasma iPTH was higher in patients with osteomalacia than in patients without osteomalacia (P less than 0.01) or controls (P less than 0.01). Urine cAMP was higher in patients with osteomalacia than in patients without osteomalacia (P less than 0.001) or controls (P less than 0.01). TmP/GFR was significantly lower in patients with osteomalacia than in controls (P less than 0.05) but not significantly different from patients without osteomalacia. The findings of this study indicate that hyperparathyroidism occurs in a substantial proportion of patients with the osteomalacia of chronic liver disease. Moreover, osteomalacia in chronic liver disease is clearly related to reduced levels of plasma 25-hydroxyvitamin D. We conclude that hepatic osteomalacia is a vitamin D deficiency state and there is no need to suggest an unusual aetiology. PMID- 6274547 TI - Lack of suppression of insulin secretion by exercise in patients with insulinoma. AB - Serum glucose, insulin and glucagon concentrations were measured in three patients with insulin-producing tumours of the pancreas while performing an exercise test. In contrast to the normal adrenergic inhibition of insulin release in response to exercise, plasma insulin concentration remained at a constant and high level during exercise in patients with insulinomas. Their plasma glucose concentrations fell during exercise and in the post-exercise period. No significant changes occurred in plasma glucagon concentration. An exercise test may be a useful new diagnostic tool in organic hyperinsulinism. PMID- 6274548 TI - [Hormone receptor diseases (author's transl)]. PMID- 6274549 TI - Biochemical aspects of gastric secretion. PMID- 6274550 TI - Pancreatic exocrine secretion. PMID- 6274551 TI - Hepatic fibrosis. PMID- 6274552 TI - Platelets, prostaglandins and thrombotic disorders. PMID- 6274553 TI - Eleven years experience of oral urea treatment in liver malignancies. PMID- 6274555 TI - Solitary midbrain metastasis. AB - The available clinical and pathological data of 5 cases with solitary midbrain metastasis including 2 of the present study are reviewed. Progressive dementia occurred in one case and mild dementia in another who also developed ocular symptoms. Ocular symptoms with sensory and coordination disturbances were seen in one, and only ocular symptoms in another case. Right-sided hemiplegia of 5 years duration occurred in the remaining case. Survival in tegmentum lesions is short. PMID- 6274554 TI - Myasthenia gravis. A survey. AB - Myasthenia gravis is a chronic disease characterized by a fluctuating weakness of voluntary muscle, with a preference for the muscles innervated by cranial nerves. The pathophysiological mechanism is a loss of postsynaptic acetylcholine receptors to less than 20-30% so that the safety margin of neuromuscular transmission is lost. It is probable that the function of the remaining acetylcholine receptors is impaired by antibodies against receptor protein, which can be demonstrated in the serum in 80-90% of the patients, and which are highly specific for the disease. An experimental autoimmune myasthenia can be induced in many animal species by immunization with purified receptor protein and this disease is remarkably similar to the human myasthenia with exception of the fluctuating course. The human disease has to be considered as an autoimmune disease, although the initiating mechanism is unknown. The occurrence of tumors of the thymus in 10-15% and the presence of germinal centres in about 70% of the thymus glands removed by operation are highly suggestive of the importance of the thymus in the pathogenesis, but the definite mechanism (harbouring of an abnormal antigen in myoid cells, or/and false instruction of thymocytes with lack of suppressor cells) is essentially unknown. In most patients the disease tends ot have a favourable course from 5-10 years after onset and complete remission occur in about 20% after 10-20 years. Therapy with anticholinesterases, providing an increase in acetylcholine, is of partial benefit in most patients. Thymectomy has an excellent effect in about 30% of the patients without thymoma under the age of 40 during the first three years of the disease, and is of benefit in still another 30-40%. The use of prednisone and immuno-suppressive drugs has improved the prognosis of the 20% of the patients with severe life threatening symptoms, half of whom have a thymoma. PMID- 6274556 TI - Meige's syndrome: clinical, pharmacological and radiological observations. AB - The clinical pharmacological, and neuroradiological observations in six patients with spontaneous blepharospasm-oromandibular dystonia (Meige's) syndrome are recorded. This group consisted of five males and one female, mean age at onset being 50.3 years. The duration of symptoms ranged from three months to 12 years, three patients having had symptoms for over four years. The dyskinesia was arrhythmic and asymmetrical in the orbicularis oculi and masseter muscles electrophysiologically. Pharmacological studies evinced no consistent response to parenteral physostigmine, no response to oral levodopa and no significant improvement in the dyskinesia following oral haloperidol. Lumbar air encephalogram was done in five patients, and showed frontal cortical atrophy without ventricular dilation in three. It is concluded that Meige's syndrome is a distinct nosological entity, and that physostigmine test is unlikely to be helpful in the differential diagnosis from neuroleptic-induced tardive dyskinesia. Neurotransmitter imbalance in the basal ganglia in this disorder remains to be established, and at present there is no satisfactory drug treatment for this progressively disabling movement disorder. PMID- 6274557 TI - Neurologic manifestations of homocystinuria. AB - Recognition of the disease homocystinuria by the neurologist may be of importance as is illustrated in the dramatic case history of a 20 year old woman. Confusion with Marfan's syndrome frequently occurs. The clinical and pathologic anatomical features are described. A review of the literature is given and the use of contraceptives is discussed. PMID- 6274558 TI - Relation between EEG and disability scores in multiple sclerosis. AB - A disability-scoring and an EEG-spectral analysis was performed in patients suffering from clinically and biochemically (oligoclonal aspect in cerebro-spinal fluid) proven multiple sclerosis at two stages of their illness, that are before and after a short intensive immuno suppressive therapy. In both stages a relationship is found between the degree of disability and the amount of beta activity in the fronto-central area and the amount of the theta activity in the temporal area. Significant improvement in the clinical state of the patient and a marked increase of the mean alpha frequency in the parietooccipital region could be demonstrated after short intensive immunosuppressive therapy. PMID- 6274559 TI - Metastatic meningioma. PMID- 6274560 TI - Archaeology: the big neurosurgical dig. PMID- 6274561 TI - Medical and surgical management of functional pituitary tumors. PMID- 6274562 TI - Mecillinam in cerebrospinal fluid in children. AB - The penetration of mecillinam, a new beta-lactam penicillin-like antibiotic, into cerebrospinal fluid (CSF) was investigated in 11 children in whom all but 1 was presumed to have non-inflamed meninges. 30 minutes after a single intravenous dose of 30mg/kg, a concentration of approximately 0.30 micrograms/ml was achieved in the CSF (corresponding mean serum concentration 75 micrograms/ml), and this level was maintained during the next 4 hours in spite of rapidly declining serum concentrations, indicating a slow equilibration of mecillinam over the blood liquor barrier. 1 child with suspected inflamed meninges showed a much higher CSF level of 12.1 micrograms/ml. It is concluded that mecillinam, as is the case for other penicillins, apparently crosses non-inflamed meninges poorly. PMID- 6274563 TI - Factors influencing cardiac hypertrophy in hypertensive patients. AB - 1. Seventeen male patients with essential hypertension were studied after 4 weeks of placebo and after 8 weeks of beta-adrenoceptor-blockade therapy with atenolol (100 mg/kg). 2. The influence of the following factors on left ventricular wall thickness and left ventricular mass index as determined by echocardiography was examined: patient's age, duration of hypertension, arterial pressure, blood pressure variability, supine heart rate, maximal exercise heart rate, left ventricular wall stress and 24 h urinary catecholamines. 3. Left ventricular mass index was related to systolic blood pressure (r = 0.54, P less than 0.05) and to extent of increase in heart rate with maximal exercise (r = 0.62, P less than 0.05). No significant correlation was present between mass index and other variables. 4. After atenolol therapy, left ventricular mass index decreased by 14 g/m2 (12%). Changes in mass were related to its initial value (r = 0.69, P less than 0.01) and to % change in wall stress (r = 0.64, P less than 0.05). Patients who had a decrease in mass index of 10% or greater had an initially lower diastolic pressure (P less than 0.001). Other factors did not appear to influence significantly the regression of hypertensive left ventricular hypertrophy. PMID- 6274564 TI - Ki+-Nao+ countertransport in erythrocytes of patients with essential hypertension. AB - 1. We describe in this paper a new ouabain-insensitive pathway for Na+ and K+ in human erythrocytes. K+ efflux was measured in cells loaded by the p chloromercuribenzene-sulphonate (PCMBS) procedure to contain approximately equal amounts of Na+ and K+. K+ efflux was stimulated by external Na+ in the presence of ouabain and frusemide. Na+-stimulated K+ efflux was 0.35 +/- 0.12 (mmol h-1 l 1 of cells) in eight normal subjects and 0.64 +/- 0.13 in 13 patients with essential hypertension. 2. The Na+-stimulated K+ efflux was not observed in cells loaded in the presence of EGTA. This inhibition by EGTA suggests that K+ efflux is dependent on intracellular calcium. The Ki+-Nao+ countertransport of hypertensive patients was also inhibited by EGTA. The elevated K+-Na+ countertransport of hypertensive patients could be due to a higher intracellular Ca2+ content (Ca2i+) or to an increased affinity for Cai2+. The relationship of this pathway to the Gardos effect is not clear since Na+-stimulated K+ efflux occurs without metabolic depletion or inhibition of the Ca2+ pump. As a tentative hypothesis, we relate the Ca2+-dependent downhill movement of K+ and Na+ to the Ca2+-dependent channels described in muscle and nerve, but other hypotheses cannot be excluded at this stage of our research. PMID- 6274565 TI - Elevated plasma adrenaline reflects sympathetic overactivity and enhanced alpha adrenoceptor-mediated vasoconstriction in essential hypertension. PMID- 6274566 TI - An increase in a circulating inhibitor of Na+,K+-dependent ATPase: a possible link between salt intake and the development of essential hypertension. AB - 1. The plasma's ability to stimulate guinea-pig renal glucose 6-phosphate dehydrogenase (G6PD) in vitro was measured by a cytochemical technique in 23 normotensive subjects and 19 patients with hypertension, all of whom were studied on their normal sodium intake. The ability of plasma to stimulate renal G6PD was significantly (P less than 0.001) increased in the hypertensive patients (mean 195 +/- 52 units/ml) compared with the normotensive subjects (mean 22.2 +/- 5.8 units/ml). In all 42 individuals, there was a significant correlation between diastolic pressure and the ability of plasma to stimulate G6PD (r = 0.69 P less than 0.001). 2. The ability of plasma to stimulate G6PD was greatest in the hypertensive patients with values of plasma renin activity below the normal range. In the normotensive subjects the ability of plasma to stimulate G6PD was significantly greater in the older subjects. 3. As the ability of plasma to stimulate G6PD reflects its ability to inhibit Na+,K+-dependent ATPase, these results suggest that patients with essential hypertension have an increase in a circulating inhibitor of Na+,K+-ATPase. The results support the hypothesis that a rise in a circulating sodium transport inhibitor may, in part, be responsible for the rise in blood pressure in essential hypertension, and may form the link between salt intake, abnormalities of sodium transport and a rise in blood pressure. PMID- 6274567 TI - Contribution of cardiovascular reflexes to differences in beta-adrenoceptor mediated responses in essential hypertension. PMID- 6274568 TI - Endogenous agonist regulation of alpha-adrenoceptors in man. AB - 1. Plasma noradrenaline concentrations and platelet alpha-receptor properties were measured in 14 normal subjects, five phaeochromocytoma patients and nine patients with multiple system atrophy and sympathetic nervous system degeneration (MSA). 2. Plasma noradrenaline concentrations in phaeochromocytoma were 10- to 20 times greater than those of normal subjects and one-fifth to one-half of normal in MSA patients. 3. Platelet alpha-receptor numbers were increased in MSA and decreased in phaeochromocytoma compared with normal subjects. There was no change in platelet alpha-receptor affinity in either group. 4. The inverse relationship between alpha-receptor numbers and plasma noradrenaline concentrations may reflect regulation of alpha-receptor number by endogenous adrenergic agonist concentrations in man. PMID- 6274569 TI - Distribution and properties of angiotensin-converting enzyme in cerebral microvessels of the rabbit and rat. PMID- 6274570 TI - Blood pressure, renal and metabolic effects of ACTH in normotensive man. AB - 1. The blood pressure, renal and metabolic effects of adrenocorticotropic hormone (ACTH) have been studied in six normotensive subjects and two patients with Addison's disease on maintenance steroid therapy. 2. In normotensive subjects, 5 days ACTH treatment (0.5 mg 12 hourly) was associated with a rise in systolic blood pressure and mean arterial pressure. There was a small rise in diastolic pressure but no consistent change in heart rate. Plasma sodium increased and plasma potassium fell. Serum creatinine and urea concentrations were unchanged. Fluid intake increased and urine output was unchanged but ACTH withdrawal was associated with a diuresis. There was an initial reduction in urinary sodium excretion and a natriuresis after ACTH withdrawal. Plasma volume and body weight rose. 3. ACTH produced increases in plasma cortisol, 11-deoxycortisol, corticosterone, deoxycorticosterone, aldosterone, 17 alpha-hydroxyprogesterone and 17 alpha,20 alpha-dihydroxyprogesterone. Plasma renin concentration fell. 4. Patients with Addison's disease showed no change in blood pressure or in any other metabolic variable studied. 5. The effects of ACTH in man resembled those found in sheep. PMID- 6274571 TI - ACTH stimulates plasma renin and angiotensin II in man. AB - 1. Adrenocorticotropic hormone (ACTH; 10 i.u./day) was infused for 34 h into normal male subjects. Some subjects were additionally treated with propranolol or indomethacin. Others received sham infusions or hydrocortisone infusions instead of ACTH. 2. ACTH, but not sham or hydrocortisone infusions, led to a significant increase in plasma renin activity and angiotensin II concentration with a lag period of 7--10 h and a maximum response after 24 h. ACTH may be a physiological regulator of renin secretion, perhaps through a 'trophic' effect on the juxtaglomerular apparatus. 3. The effect of ACTH on renin is not mediated by a rise in plasma renin substrate, probably not by renal beta-adrenoreceptors, but perhaps by prostaglandins. 4. A dissociation between plasma cortisol and aldosterone during ACTH infusion suggests that ACTH, in this dosage, stimulates aldosterone on the second day through renin and angiotensin II, before its secretion is finally suppressed during more prolonged infusion. PMID- 6274572 TI - Effects of the oral angiotensin-converting enzyme inhibitor MK-421 in human hypertension. AB - 1. The effects of the new oral angiotensin-converting enzyme (ACE) inhibitor MK 421 on blood pressure, plasma renin activity, angiotensin-II, aldosterone and converting enzyme activity were assessed in 16 hypertensive patients followed for 6--18 weeks. 2. After an initial titration period, maintenance doses of 2.5--40 mg once daily produced satisfactory blood pressure control in 11 and a partial but significant decrease in the remainder. 3. Treatment was associated with no change in heart rate and no orthostatic hypotension. At 24 h after the first effective dose, blood pressure was still decreased, plasma ACE was suppressed to 16% of the baseline, plasma angiotensin to 58%, aldosterone to 42%, and plasma renin activity was elevated to 228% of the baseline. 4. Magnitude of blood pressure fall was significantly correlated with the height of pretreatment blood pressure but not with baseline levels of plasma renin activity. PMID- 6274573 TI - Effect of converting enzyme inhibition on the systemic and renal responses to acute isotonic volume expansion in normal men. PMID- 6274574 TI - Alpha 1-Adrenoceptor blockade: dissociation of its effects on renin release and arterial blood pressure in man. AB - 1. The possibility that the juxtaglomerular alpha 1-adrenoceptors mediate an inhibitory action on renin release in man was examined in seven patients with essential hypertension, by measuring (i) the acute effects of prazosin (0.25 mg intravenously), a selective alpha 1-adrenoceptor-blocking agent, on arterial pressure and plasma renin activity, the degree of alpha-blockade induced by the drug being assessed by comparing the pressor response with that to a test dose of phenylephrine before and after prazosin administration, and (ii) the increases in plasma renin activity in response to isoprenaline before and during the prazosin induced alpha-blockade. 2. Twenty minutes after the infusion of prazosin, when the pressor response to phenylephrine was reduced by 80% with respect to control, (i) mean arterial pressure was practically unchanged, (ii) plasma renin activity was almost doubled and (iii) the increases in plasma renin activity in response to isoprenaline were significantly greater, both in absolute and percentage values, than those observed before prazosin. 3. The increments in baseline plasma renin activity induced by prazosin in the absence of decrease in arterial pressure and the enhancement in renin responsiveness to the beta-adrenoceptor stimulus suggest that, in man, the juxtaglomerular alpha 1-adrenoceptors exert a direct, suppressive action on renin release. PMID- 6274575 TI - alpha-Adrenoceptors and age-dependent decrease in plasma renin activity in essential hypertension. PMID- 6274576 TI - Beta-Endorphin, an endogenous depressor agent in the rat? AB - 1. The role of opiates in cardiovascular regulation has been investigated. 2. In urethane-anaesthetized renal hypertensive rats (two-kidney, one-clip Goldblatt model), intracerebroventricular beta-endorphin (10 microgram) caused a greater fall in mean arterial pressure than in sham-operated controls. 3. Unilateral injection of beta-endorphin into the nucleus tractus solitarii of the urethane anaesthetized rat resulted in a U-shaped dose--response relationship, with a fall in mean arterial pressure and heart rate occurring at low doses. Doses above 10 ng caused a rise in pressure, accompanied by a variable effect on heart rate. 4. The fall in blood pressure and heart rate was prevented by prior subcutaneous administration of naloxone. Naloxone caused an increase in blood pressure when administered alone. 5. These results suggest a depressor role of an endogenous brain opiate, possibly beta endorphin; a site of action is probably the nucleus tractus solitarii. PMID- 6274577 TI - Effects of bromocriptine on blood pressure and plasma beta-endorphin in spontaneously hypertensive rats. AB - 1. Immunoreactive beta-endorphin (IR-beta EP) was two- to three-fold higher in pituitary neuro-intermediate lobes (N-IL) of spontaneously hypertensive rats (SHR) than of normotensive Wistar--Kyoto (NT-WKY) controls. 2. Plasma levels of IR-beta EP were lower in SHR than in NT-WKY rats. 3. Intravenous injections of morphine lowered blood pressure of both SHR and NT-WKY rats to the same level; naloxone restored blood pressure of both groups to pre-morphine values. 4. Infusion of bromocriptine in SHR for 1 week lowered blood pressure and N-IL IR beta EP concentration. 5. These results confirm and extend postulated dopaminergic defect in this model of hypertension. PMID- 6274578 TI - Decrease in alpha-adrenoceptor-mediated vasoconstriction parallels the antihypertensive response to propranolol in patients with normal renin essential hypertension. PMID- 6274579 TI - Membranes, ions and hypertension. PMID- 6274580 TI - Haemodynamic effects of noise exposure before and after beta 1-selective and non selective beta-adrenoceptor blockade in patients with essential hypertension. AB - 1. Noise stimulation (100 dBA) for 10 min caused a significant increase in diastolic (7.0%, P less than 0.001) and mean arterial blood pressure (4.3%, P less than 0.01) in patients with essential hypertension. 2. The blood pressure response to noise was due to an increase in total peripheral resistance (4.8%, P less than 0.02); heart rate, stroke volume and cardiac output were unchanged. 3. beta 1-selective adrenoceptor blockade (metoprolol) did not change the haemodynamic reaction pattern induced by noise. 4. Noise exposure during non selective beta-adrenoceptor blockade (propranolol) caused an accentuated blood pressure response with increments of both systolic and diastolic blood pressure as well as a more pronounced rise in total peripheral resistance. 5. The haemodynamic changes induced by noise stimulation at 100 dBA totally disappeared after 5 min of quiet rest at 40 dBA. PMID- 6274581 TI - Management of thallium poisoning. PMID- 6274582 TI - Management of small cell bronchogenic carcinoma. PMID- 6274583 TI - Computer programming for nucleic acid studies. II. Total chemical shifts calculation of all protons of double-stranded helices. AB - A FORTRAN computer program called SHIFTS is described. Through SHIFTS, one can calculate the NMR chemical shifts of the proton resonances of single and double stranded nucleic acids of known sequences and of predetermined conformations. The program can handle RNA and DNA for an arbitrary sequence of a set of 4 out of the 6 base types A,U,G,C,I and T. Data files for the geometrical parameters are available for A-, A'-, B-, D- and S-conformations. The positions of all the atoms are calculated using a modified version of the SEQ program [1]. Then, based on this defined geometry three chemical shift effects exerted by the atoms of the neighboring nucleotides on the protons of each monomeric unit are calculated separately: the ring current shielding effect: the local atomic magnetic susceptibility effect (including both diamagnetic and paramagnetic terms); and the polarization or electric field effect. Results of the program are compared with experimental results for a gamma (ApApGpCpUpU) 2 helical duplex and with calculated results on this same helix based on model building of A'-form and B form and on graphical procedure for evaluating the ring current effects. PMID- 6274584 TI - Hepatitis A, B and N: some practical aspects. PMID- 6274586 TI - The nervous system: a communication's symphony. PMID- 6274585 TI - Studies on ovarian and adrenal studies at different phases of the menstrual cycle. IV. The effect of dexamethasone suppression and subsequent ACTH stimulation at different phases of the menstrual cycle and following the administration of 150 mg of depot-medroxy-progesterone acetate (DMPA). AB - In an attempt to assess the effect of depot-medroxyprogesterone acetate (DMPA) on adrenal function, to peripheral levels of a variety of steroids were estimated after dexamethasone suppression and subsequent ACTH stimulation in 5 subjects. The steroid responses observed 16 and 54 days after a single injection of 150 mg of DMPA were compared with those obtained in the same women before DMPA treatment or with those found at different phases of the cycle in another group of 6 normally menstruating women. The levels of cortisol, pregnenolone, 17 hydroxypregnenolone, dehydroepiandrosterone, androstenedione and dihydrotestosterone were significantly suppressed by dexamethasone and stimulated by ACTH in all phases of the cycle, and before and after DMPA administration. The extent of suppression or stimulation (expressed as a percentage) was different in case of different steroids, but was rather constant with regard to the same steroid. Hence, the functional capacity of the adrenal as reflected by the levels of these steroids. Dexamethasone administration did not exert any influence on the levels of progesterone and 17-hydroxyprogesterone during the different phases of the cycle; however, ACTH administration increased the levels of these two steroids in the follicular phase and also after DMPA treatment. The levels of estradiol and lutropin were not influenced by dexamethasone or ACTH at any of the occasions studied. It is concluded that the functional capacity of the adrenal cortex is constant during the various cycle phases and is not affected by the administration of a single dose of 150 mg of DMPA. PMID- 6274587 TI - Antibody mediated destruction of keratocytes infected with herpes simplex virus. AB - The ability of complement-dependent antibody to mediate destruction of rabbit corneal cells infected with the RE and KOS strains of HSV-1 was examined. Three sources of target cells were used: BHK-21, a monitor of surface antigen expression and cell lysis; SIRC, a continuous cell line of rabbit corneal origin; stromal keratocytes, grown from the middle layer of excised rabbit corneas. Rabbits were infected intrastromally with the RE strain, which causes a high incidence of stromal keratitis, and sacrificed at designated times. Results showed that all three cell types, infected with either HSV strain, expressed surface antigens and were susceptible to lysis by antibody produced during the course of stromal disease. 51Cr-release assays showed that cytolytic antibody appeared as early as 5 days postinfection and reached maximum by day 14-20. Findings suggest a direct relationship between the in vitro demonstration of complement dependent cytolysis of infected stromal cells and the disease process in experimental animals. PMID- 6274588 TI - Increased ability of human embryonic fibroblasts to accumulate Ca2+ due to cytomegalovirus infection. AB - Cytomegalovirus infection of fibroblasts enhances the ability of these cells to accumulate Ca2+. This functional alteration precedes the development of the cytopathic effect, and it is proposed that increased intracellular Ca2+ concentration may initiate the rounding phenomenon. Changes in the aggregative behaviour and the antigenicity of the infected cells were seen to accompany this functional alteration. PMID- 6274589 TI - Reversible ventilation and perfusion abnormalities in unilateral obstructed lung. AB - An intraluminal carcinoid tumor obstructing the left mainstem bronchus produced hypoxemia through alteration in ventilation/perfusion matching. Studies of regional lung function using 133-xenon (133Xe) and a multiprobe computerized instrumentation system documented a reduction of perfusion to 22 percent and ventilation to 6 percent of the total. There was negligible washout of intravenously injected 133Xe from the left lung consistent with air trapping. Four days after left mainstem bronchial sleeve resection, perfusion, ventilation and washout of injected xenon had significantly improved and by four months postresection, all measurements were virtually normal, although complete restoration of perfusion in relation to ventilation was delayed. Regional lung function studied with a multiprobe system in this patient provided a clinical model for the study of ventilation and perfusion inter-relationships in large airway obstruction and demonstrated that a prolonged time may be required for return of perfusion to normal. PMID- 6274590 TI - Measurement of surface temperature in lung cancer. AB - We measured the surface temperature of the tumors in 27 patients with primary lung cancer, using a flexible bronchofiberscope and a thermometer made from chromel-constantan. The thermometer was inserted into the aspiration channel of the bronchofiberscope, and temperature was studied under bronchofiberscopic located in the central portion of the lung were higher than those of the carina of the trachea (P less than 0.01), truncus superior (P less than 0.01), or truncus inferior (P less than 0.05), and almost equal to those of the peripheral lung. Thus, lung cancer showed a significantly higher surface temperature. When 67Ga scintigraphy was performed in 22 patients with lung cancer, the temperature of the high-uptake group was higher than that of the low-uptake group. PMID- 6274591 TI - Anticomplement immunoenzymatic method of detecting Epstein-Barr nuclear antigen in nasopharyngeal carcinoma cells and normal epithelial cells. PMID- 6274592 TI - [Fascial closure of median laparotomies with a synthetic, resorbable suture material (polyglycolic acid)]. AB - In a prospective randomized study polyglycolic acid sutures (PGA) and polyester sutures for closure of median abdominal fascial incisions are compared. Occurrence of infection, incisional hernia, wound dehiscence and suture fistulas is evaluated. With regard to these complications no significant differences could be detected. PGA shows its advantage in less tissue reaction (avoiding fistulae). PMID- 6274593 TI - [X-ray diagnosis of small cell carcinoma of lung (author's transl)]. PMID- 6274594 TI - [Clinical analysis of 49 cases of bronchiolo-alveolar carcinoma (author's transl)]. PMID- 6274595 TI - An immunodeficient child with inflammatory bowel disease: involvement of cyclic nucleotides and effects of lithium. AB - A 3-year-old male with inflammatory bowel disease and hypogammaglobulinemia was found to have decreased T lymphocyte function. His serum was shown to depress normal T cell proliferative responses to phytohemagglutinin. Incorporation of lithium chloride to in vitro cultures enhanced autologous lymphocyte responses to phytohemagglutinin. Since lithium acts by inhibiting cAMP production, the child's lymphocytes were postulated to have increased levels of cAMP. Both lymphocytes and serum were shown to contain elevated levels of cAMP. In vivo therapy with lithium citrate was initiated and enhanced T cell numbers and function were observed concomitantly. Serum cAMP was also reduced to normal levels. The patient showed initially marked clinical improvement as assessed by mood, weight gain, and diminution of diarrhea. This clinical improvement was unfortunately not sustained despite the continued improvement in immune parameters and cAMP levels. PMID- 6274596 TI - Urinary catecholamines, plasma renin activity and blood pressure in newborns: effects of narcotic withdrawal. AB - Blood pressure and plasma renin activity were studied in 13 infants of drug dependent mothers (heroin and/or methadone) and 20 control newborns for a period of 21 days. Urinary excretion of epinephrine and norepinephrine were also measured at 24 and 48 h of age. The systolic blood pressure was found to be significantly elevated in newborns of drug-dependent mothers compared with controls (p less than 0.01), but diastolic blood pressure levels were similar. Plasma renin activity was significantly elevated (p less than 0.001) at 48-72 h and urinary epinephrine levels were significantly lower (p less than 0.05) at 24 48 h of age in the drug group, and norepinephrine levels were equivalent. Systolic hypertension and elevated plasma renin activity in these newborns of dependent mothers may be due to increased beta-adrenergic receptor sensitivity. PMID- 6274597 TI - [cGMP subunit structure of the phosphodiesterase in the outer segments of the retinal rods]. PMID- 6274598 TI - [Pharmacological data on the existence of opiate receptors in the myocardium of warm-blooded animals]. PMID- 6274599 TI - [Nature of the cell targets of the action of the antitumor preparation methylnitrosourea]. PMID- 6274600 TI - [Kidney tissue phosphoprotein phosphatase and the regulation of glutamate dehydrogenase activity]. PMID- 6274601 TI - [Isolation and identification of the matrix RNA that codes the polypeptide precursor of beta-lipotropic and corticotropic hormones. The synthesis of complementary DNA]. PMID- 6274602 TI - [Translation initiation factor activity in Krebs-2 cells infected with the encephalomyocarditis virus]. PMID- 6274603 TI - Neurochemical and behavioural correlates of the interaction between amphetamine and delta 9-tetrahydrocannabinol in the rat. AB - A single dose of delta 9-tetrahydrocannabinol (THC) (10 or 100 mg/kg) was administered via stomach gavage following the single injection intraperitoneally of an acute dose of amphetamine (1 or 3 mg/kg) in the rat. The THC effects, which included a reduction in spontaneous locomotor activity, the development of prostration, and circling behaviour, were antagonized in a dose-dependent manner by pretreatment with amphetamine. Although no change in the endogenous levels of noradrenaline, dopamine and serotonin in hypothalamus, hippocampus, caudate putamen or the rest of brain occurred with THC, it appears that THC may have increased dopamine turnover in the brain. The dose-dependent amphetamine antagonism to the behavioural effects of THC was paralleled by a dose-related increase in brain dopamine concentration. This elevation in dopamine levels may be instrumental in triggering the neuronal events responsible for the amphetamine antagonism of the actions of THC. The subtle interaction of THC with brain dopamine neural systems may be involved in the reinforcing properties of the self administration of THC. PMID- 6274604 TI - Modification by hyperoxia of chlorphentermine- or phentermine- induced effects on newborn rat lung morphology and metabolism. AB - Treatment of newborns with 20 mg/kg/day chlorphentermine orally for 1 week increased incorporation of thymidine into lung DNA without an associated change in tissue morphology or cyclic AMP levels. An increase in chlorphentermine dose to 60 mg/kg resulted in an accumulation of alveolar hypertrophic macrophages and a rise in incorporation of thymidine into lung DNA; however, cyclic AMP levels were decreased. In contrast, 20 or 60 mg/kg/day for 1 week phentermine-induced depression in the incorporation of thymidine into pulmonary DNA was accompanied by a decrease in cyclic AMP but no apparent alteration in tissue morphology. Hyperoxia did not modify the phentermine-induced changes in cyclic AMP levels and pulmonary ultrastructure. In contrast, hyperoxia altered the responsiveness of newborns to 20 mg/kg chlorphentermine as evidenced by the presence of foam cells. Data suggest that the chlorphentermine-induced increase in DNA synthesis in newborn lung seems independent of changes in cyclic AMP and tha modification of drug-induced alterations by hyperoxia may be related to the chemical structure of a compound. PMID- 6274606 TI - Aspiration cytology in the diagnosis of liver cancer. PMID- 6274607 TI - Photochemistry of phenylurea herbicides and their reactions in the environment. PMID- 6274605 TI - Adenosine and guanosine 3',5' cyclic monophosphate phosphodiesterase activities in rat small and large bowel following single and multiple exposure to 1,2 dimethylhydrazine. AB - The hydrolytic activities of adenosine 3',5'-cyclic monophosphate (cAMP) and guanosine 3',5'-cyclic monophosphate (cGMP) phosphodiesterases (PDE) in rat large and small bowel tissue were determined after exposure to the colon carcinogen 1,2 dimethylhydrazine (DMH). Immediate increases in the cAMP-PDE activities were found in the tissues after a single exposure to the chemical which returned towards normal while chronic exposure resulted in no visible changes. The increased cAMP-PDE activities may be the factor responsible for the diminished intracellular cAMP concentrations found after exposure to the carcinogen. In contrast, the cGMP-PDE activities changed little in the colon and increased in the small bowel, suggesting that the determinant for the increases in the concentration cGMP was the activation of guanylate cyclase enzymes following exposure to DMH. Ratios of cAMP to cGMP and cAMP-PDE to cGMP-PDE in the colon showed major changes after carcinogenic insult by the DMH, suggesting that such measurements might serve as useful markers for exposure to environmental toxicants. PMID- 6274608 TI - [Fatigue and depression (author's transl)]. AB - Fatigue is usually stated as a major symptom of the depressive condition. Its diagnosis value seems higher in mild depressions, that in severe depressive syndromes. In this later case the symptom is overwhelmed by psychic pain. Contrary to the common viewpoint fatigue does not seem a consequence of sadness and lack of interest. It seems we have to make a distinction between the fatigue related to anxiety and hypochondria and an other kind of fatigue that is the subjective aspect of motor and mental retardation. Its relationships with circadian rhythms and nor-adrenergic transmission are probably the best ways to explore the biological mechanisms. PMID- 6274609 TI - [Mianserin, presynaptic receptor and biochemical model of depression of Tissot (author's transl)]. PMID- 6274610 TI - [Are antidepressants different? (author's transl)]. AB - There are different groups of antidepressants with different mechanisms of action. However, by different mechanisms all these drugs are able to activate beta-adrenergic receptors, and the antidepressant effect could be related to this activation. Thus, the therapeutic effect of antidepressants is probably qualitatively similar but drugs can differenciate by the onset of action, by other effects (stimulant or sedatif) or by side-effects. PMID- 6274611 TI - Specific thyroxine nuclear receptors: DEAE-sephadex chromatography of rat liver nuclear extract. AB - Specific binding of thyroxine to protein fraction obtained by DEAE-Sephadex chromatography of dialyzed (0.4 mol 1-1 KCl) nuclear extract of purified rat liver nuclei was studied. A procedure for several-fold purification of thyroxine nuclear receptor included isolation of purified rat liver nuclei (by centrifugation through 1.7 mol 1-1 sucrose), extraction, dialysis, Sephadex G-100 and DEAE-Sephadex A-50 chromatography. It was found that a protein peak eluting at 0.05 mol 1-1 NaCl does not show any fraction specifically binding thyroxine. However, specific binding of nuclear receptor for thyroxine was found in a peak eluting at 0.20 mol 1-1 NaCl, the capacity being about 600 fmol mg-1 protein. Moreover, total binding capacity of intact rat liver homogenate, nuclei, non dialyzed and dialyzed (0.4 mol 1-1 KCl) nuclear extract, Sephadex G-100 and DEAE Sephadex A-50 chromatography protein fractions for thyroxine was tested. A marked increase (about 20 fold) in the total thyroxine binding expressed in fmol mg-1 of protein in DEAE-Sephadex A-50 eluate was found as compared to whole nuclei, nuclear extract or Sephadex G-100 chromatography fractions. Partial fractionation of rat liver nuclear extract combined with thyroxine binding studies demonstrated that rat liver nuclei contain a non-histone protein fraction--a receptor that is responsible for a specific binding of thyroxine. PMID- 6274612 TI - Seasonal changes in pituitary-adrenal reactivity in hibernating spermophiles. AB - Basal level of 11-hydroxycorticosteroids in plasma was measured during one year period in spermophiles and correlated with a physiological state of the animals. Maximum level was found in Spring after the animals had awaken from hibernation, while a minimum level was observed during hibernation. During the waking up process the level of corticosteroids increased slowly which may show that this process does not act as a stressor. In contrast, the falling asleep process starts under a decreased level of corticosteroids in plasma and is accompanied by a further decrease. Maximum stimulatory effect of adrenaline and isoprenaline on pituitary-adrenal axis was found after waking up in Spring. It was considerably less in Summer, while no such effect was found before hibernation period. However, at the same time the sensitivity of adrenals to ACTH was sufficient. From this it may be concluded that seasonal changes in pituitary-adrenal response to catecholamines may be due to changes in central regulatory mechanisms. PMID- 6274613 TI - Adrenergic mechanisms involved in the control of pituitary-adrenal activity in the rat: a beta-adrenergic stimulatory mechanism. AB - Epinephrine or isoproterenol was infused into a lateral tail vein of female Wistar rats under Nembutal anesthesia. After 20 min of diffusion, trunk blood was collected for the determination of plasma corticosterone (B) and ACTH immunoreactivity (ACTHi). Infusion of l-epinephrine resulted in a dose-related increase in plasma ACTHi and B. Maximal levels were similar to those observed during ether stress. The pituitary-adrenal system appeared more sensitive than the cardiovascular system to epinephrine, since the ED50 values of epinephrine for its effects on ACTHi and heart rate were 165 and 840 ng/kg . min, respectively. The effect of epinephrine on pituitary-adrenal activity could be mimicked by the beta-adrenergic agonist l-isoproterenol and could be blocked by the beta-adrenergic antagonist l-propranolol, whereas d-propranolol was ineffective. The response of the pituitary-adrenal system to epinephrine was not caused by effects on peripheral parameters such as the distribution or clearance of ACTH or B but was mediated by an increase in ACTH release. The pituitary adrenal response to epinephrine and isoproterenol was not related to changes in heart rate, blood pressure, or vasopressin secretion. Infusion of epinephrine at a dose that induced a maximal increase in plasma ACTHi and B (1000 ng/kg . min) resulted in a circulating epinephrine concentration of 11 pmol/ml, which is within the physiological range. From these data we conclude that 1) circulating epinephrine can stimulate pituitary-adrenocortical activity, 2) this action is mediated by a beta-adrenergic receptor mechanism, and 3) such a mechanism may be involved in the response of the pituitary-adrenal axis during certain forms of stress. PMID- 6274614 TI - Effects of thyrotropin, acetazolamide, 4-acetamido-4'-isothiocyanostilbene-2,2' disulfonic acid, perchlorate, and ouabain on the distribution of iodide ion in cells and luminal fluid of turtle thyroid. PMID- 6274615 TI - Role of lipoproteins and 3-hydroxy-3-methylglutaryl coenzyme A reductase in progesterone production by cultured bovine granulosa cells. AB - The relative contributions of lipoproteins and 3-hydroxy-3-methylglutaryl coenzyme A (HMG CoA) reductase to progesterone production by bovine granulosa cells exposed to plasma or liquor folliculi (LF) were studied. LF did not contain and very low density lipoprotein (VLDL), intermediate density lipoprotein (IDL), or low density lipoprotein (LDL). These lipoproteins were present in the plasma at concentrations of 92 micrograms protein/ml for VLDL and IDL together and 139 micrograms protein/ml for LDL. In contrast, high density lipoprotein (HDL) was present in LF at a concentration (763 micrograms protein/ml) that was 59% of that in plasma (1293 micrograms protein/ml). Bovine granulosa cells exposed to human plasma produce progesterone in response to dibutyryl cAmP. Sixty-three percent of the progesterone released by the cells was dependent on LDL but not HDL derived from human plasma. When cells were exposed to bovine plasma, 75% of the progesterone release was dependent on the presence of lipoproteins in the medium. Both LDL and HDL of bovine origin were able to support progesterone production, although LDL was effective at concentrations (on a molar basis) 20-fold lower than HDL. The LF was able to support progesterone production 45% as well as bovine plasma. The differences between the greater ability of the whole fractions and the lesser ability of their respective lipoprotein-deficient derivatives to support progesterone synthesis were 4-fold for bovine plasma, 2.7-fold for human plasma, and 1.7-fold for LF. The relative abilities of equivalent concentrations of LDL to restore the rate of progesterone synthesis seen in the lipoprotein deficient fraction toward that seen in the whole fraction were greatest in the LF, intermediate in human plasma, and least in bovine plasma. These observations taken together suggest that the low level of support of progesterone synthesis that is offered by LF is due to its deficiency in LDL. HMG CoA reductase, the regulated and rate-limiting enzyme of cholesterol synthesis, was induced (2- to 3 fold) by dibutyryl cAMP and was suppressed by both human and bovine LDL and to a lesser extent by bovine HDL. Compactin, a competitive inhibitor of HMG CoA reductase, inhibited progesterone production relatively little when cells were exposed to complete plasma or LF. However, when cells were exposed to a lipoprotein-deficient bovine plasma or LF, compactin was very efficient in reducing (by 76%) progesterone release. Bovine granulosa cells exposed to plasma primarily use cholesterol derived from LDL in order to produce progesterone. Their ability to produce progesterone when exposed to LF was limited, and the cells were probably more dependent on de novo cholesterol synthesis than cells exposed to plasma. PMID- 6274616 TI - The effects of chronic human chorionic gonadotropin treatment on Leydig cell function. AB - The purpose of this study was to examine the effect of chronic daily hCG treatment on interstitial cell function in the rat, as judged by plasma testosterone levels, the testicular binding of labeled hCG, and the capacity of the testis to respond to gonadotropin stimulation by the production of testosterone in vitro. TWenty-four hours after the first injection of 100 IU hCG there was a significant decline in hCG binding to testis homogenates and an inability to respond to hCG stimulation in vitro, After 7 days of daily injections of 10 IU or 100 IU hCG, the loss of hCG binding was maintained. However, despite the marked decline in hCG binding, there was an enhanced testosterone response to hCG stimulation in vitro, and plasma testosterone levels were significantly elevated. With continued injections of hCG for 14 or 21 days, the testes remained hyperresponsive to hCG stimulation in vitro, but hCG binding returned to control levels, and plasma testosterone concentrations declined and were not statistically different from controls. The latter changes probably result from the formation of specific hCG antibodies (Kd at 4 C, 7.8 +/- 4.5 X 10(-10) M) that were detected in plasma from rats treated for 14 or mopre days with hCG. The formation and levels of the hCG antibodies in these animals were sufficient to neutralize the effects of the exogenous hCG, thereby returning plasma testosterone levels to normal and restoring the complement of hCG receptors. PMID- 6274617 TI - Relaxin receptor in the rat myometrium: regulation by estrogen and relaxin. PMID- 6274618 TI - Effects of glucocorticosteroids and glucagon on argininosuccinate synthetase, argninosuccinase, and arginase in fetal rat liver. AB - In the present study we examined the in vivo effects of glucocorticosteroids and glucagon on the developmental formation of the individual urea cycle enzymes argininosuccinate synthetase, argininosuccinase, and arginase during the late fetal period. In particular, addition of exogenous glucagon caused a rise in argininosuccinase and arginase activities in the livers of rat fetuses at term but not at earlier stages. Glucagon produced a rise in argininosuccinase activity earlier if fetuses were previously treated with cortisol. When fetuses were deprived of corticosteroid (hypophysectomy in utero), glucagon no longer promoted the argninosuccinase activity, indicating that adrenal glucocorticoids are required for normal enhancement of the enzyme activity by glucagon. Dibutyryl cAMP was still effective in hypophysectomized fetuses. Results obtained by injected combinations of inducers indicated that a glucocorticosteroid-glucagon interaction might be involved in the regulation of argininosuccinate synthetase and argininosuccinase. No synergistic action was found on arginase activity in vivo. PMID- 6274620 TI - Effect of divalent cations on the binding of 3,5,3'-triiodothyronine to isolated rat liver nuclei. PMID- 6274619 TI - Stress-induced inhibition of the plasma corticosterone response to a subsequent stress in rats: a nonadrenocorticotropin-mediated mechanism. AB - The present study was designed to define further the relationships between ACTH and corticosterone secretion after repeated administration of a discrete restraint stress in rats. The possibility that plasma ACTH and corticosterone responses to stress may be modified by prior exposure to stress was examined in male rats using a 2-min restraint stress. The peak plasma ACTH response to a single restraint stress occurred at 2.5-5 min after the onset of the stress, and plasma ACTH returned to the basal concentration by 30 min. The plasma corticosterone concentration after this stress peaked at 15-30 min and returned to the control range by 60-90 min. The time courses of the plasma ACTH and corticosterone responses to restraint stress after administration of three prior stresses at 90-min intervals were similar to those after a single stress. Stress induced increments in plasma concentrations of ACTH and corticosterone were similar in rats that received either a single restraint stress or as many as seven stresses repeated at 90-min intervals. Next, we examined the plasma ACTH and corticosterone responses to repeated stress applied at intervals of less than 90 min (30 or 60 min), that is, at times at which the plasma corticosterone concentration had not yet returned to basal levels. The plasma ACTH responses to the second stress were similar in magnitude and duration to the response after a single stress, whether the second stress was applied 30, 60, or 90 min after the first stress. The plasma corticosterone response to a second stress applied at 90 min was identical to the response after the initial stress. In contrast, the plasma corticosterone responses to a second stress applied 30 or 60 min after the initial stress were markedly reduced. The decrease in plasma corticosterone response to the second stress did not result from a decrease in secretion of bioactive ACTH; no difference was found between the magnitude of the plasma ACTH response to the initial stress and a subsequent stress applied 30 min later using either bioassay or immunoassay measurements. Also, the rate of corticosterone catabolism was not increased by prior stress; the rate of disappearance of corticosterone from plasma was identical after an initial or a subsequent stress applied at 30 min. We were unable to demonstrate decreased adrenocortical responsiveness to ACTH after an initial stress in dexamethasone-suppressed rats; in these rats the plasma corticosterone response to exogenous ACTH was not decreased by prior restraint stress. These data clearly define a period of decreased adrenocortical response to subsequent stress after stress-induced activation of adrenocortical secretion. Furthermore, this altered adrenocortical response appears to be mediated by a nonadrenocorticotropin mechanism. PMID- 6274621 TI - Induction of cryptic lactogenic hormone binding in livers of adult female mice treated neonatally with estradiol or nafoxidine. AB - Female mice were treated with 25 micrograms of either estradiol-17 beta or the antiestrogen Nafoxidine (U11100A) or sesame oil carrier within 36 h of birth. The former treatments result in persistently elevated serum PRL levels in the adult animals. Therefore, when the mice reached maturity (3-6 months of age), their livers were examined for the level of lactogenic binding. When microsomal preparations were examined for their ability to bind either lactogenic hormone, PRL, or hGH, no differences in the control or treated groups were observed. The presence of masked or cryptic binding sites were observed. The presence of masked or cryptic binding sites was examined by either solubilization of the membrane preparations or by treatment with the methyl donor S-adenosyl-L-methionine. In all cases cryptic sites were present. In control animals unmasking of cryptic binding sites produced an increment in binding of 20-60%, but in the estradiol-17 beta- and Nafoxidine-treated animals, a 120-170% increase in binding sites was observed with no significant changes in Ka. These increases observed in the latter groups are similar to the 94-175% increase seen in livers of late pregnant and lactating mice. Thus, the persistently high serum PRL levels in neonatally treated mice result in the induction of hepatic lactogenic receptors, primarily in a masked or cryptic form. PMID- 6274622 TI - Modulation by thyroid status of hepatic low Km phosphodiesterase. AB - The effects of thyroid status on the activity of hepatic cAMP phosphodiesterase (PDE) were studied in the rat. Male rats were rendered hyperthyroid by treatment with T3 or hypothyroid by treatment with propylthiouracil. The hepatic particulate low Km PDE was solubilized, and its activity was measured at concentrations of 0.12-1.3 microM cAMP. The Km decreased in hypothyroidism and tended to increase in hyperthyroidism with respect to individual controls. The maximal velocity (Vmax) was unaffected by changes in thyroid status. The increases in Km correlated with increasing plasma T3, whereas the Vmax did not. Concentrations of cAMP increased in the livers from hyperthyroid rats and decreased in those from hypothyroid, in comparison with euthyroid rat livers. The effects of thyroid status on various aspects of hepatic lipid metabolism reported from this laboratory may, in part, have resulted from alterations in hepatic cAMP concentrations. These alterations, may have resulted secondarily from changes in the activity of hepatic PDE by changes in the Km for cAMP with little change in the Vmax. PMID- 6274623 TI - In vivo corticotropin-releasing factor-induced secretion of adrenocorticotropin, beta-endorphin, and corticosterone. AB - A 41-residue peptide purified as a corticotropin-releasing factor/beta-endorphin releasing factor (CRF) in vitro was tested for its ability to stimulate the secretion of ACTH, beta-endorphin, and corticosterone in three animal groups: 1) unanesthesized rats bearing indwelling venous cannulae, 2) rats pretreated with chloropromazine plus morphine sulfate plus pentobarbital (CPZ-MS-Nb, and 3) rats with hypothalamic deafferentiations in the frontal and lateral retrochiasmatic areas. In all three bioassays iv administration of 0.1-10 micrograms CRF elicited a dose-related increase in plasma ACTH and beta-endorphin values over a 5- to 15 min period. Corticosterone secretion was also elevated but responded maximally with all doses of CRF tested. Pretreatment of CPZ-MS-Nb animals with 20 micrograms dexamethasone 4 h before assay abolished the CRF-induced hormone secretion. These data suggest that CRF may play a physiological role in the regulation of the hypothalamic-pituitary-adrenal axis. PMID- 6274624 TI - Antagonistic interactions of adenosine and prostaglandin F2 alpha modulate acute responses of luteal cells to luteinizing hormone. PMID- 6274625 TI - Involvement of alpha 1-adrenergic receptors in the inhibitory effect of catecholamines on the thyrotropin-induced release of thyroxine by the mouse thyroid. AB - TSH enhanced the release of T4 from the mouse thyroid incubated in vitro. Norepinephrine, a nonspecific alpha-adrenergic agonist, and methoxamine, an alpha 1-agonist, inhibited the TSH-stimulated release of T4 at 10(-4) and 10(-5) M, whereas clonidine, an alpha 2-agonist, exerted a weak inhibition. The inhibitory effect of 10(-5) M norepinephrine on the T4 release stimulated by TSH was prevented by prazosin, an alpha 1-antagonist, at concentrations higher than 10( 7) M, whereas yohimbine, an alpha 2-antagonist, exerted weak activity in antagonizing the inhibition induced by norepinephrine. Norepinephrine, methoxamine, and clonidine did not significantly reduce the cAMP accumulation stimulated by TSH in the mouse thyroid incubated in vitro. These findings in the mouse thyroid indicate that catecholamines act by way of alpha 1-adrenergic receptors to suppress TSH-stimulated release of T4 without reducing the cAMP levels stimulated by TSH in the mouse thyroid. PMID- 6274626 TI - Pituitary receptor sites for gonadotropin-releasing hormone: effect of castration and substitutive therapy with sex steroids in the male rat. AB - To examine the role of pituitary gonadotropin-releasing hormone (GnRH) receptors (pit GnRH-R) in the regulation of gonadotropin secretion, male rats were orchidectomized and then selectively received substitutive therapy with sex steroids. Pituitary content of GnRH-R was determined by saturation analysis, using radioiodinated [D-Trp6,(N-Et)Pro5,des-Gly10]GnRH as tracer. Castration produced a rapid and sustained increase of the number of GnRH-R, which doubled after 2 days, and after 10 days the pituitary content of GnRH-R was 258 +/- 23 fmol/pituitary compared to 103 +/- 12 fmol/pituitary for sham-operated control animals. No change of the affinity constant (Ka) was observed (Ka = 1.13 +/- 0.08 X 10(10) M-1; n = 14). Plasma LH increased 5- to 10-fold and FSH-2- to 3-fold after castration, and hypothalamic GnRH content was depleted by 30-60%. Immediate substitution of castrated rats with testosterone propionate (250 micrograms daily) prevented the increases of both plasma gonadotropins and of GnRH-R. Treatment of acutely castrated rats for 7 days with testosterone propionate (50 200 micrograms), 5 alpha-dihydrotestosterone propionate (25-400 micrograms), or estradiol benzoate (2 micrograms) prevented the rise in pit GnRH-R in a dose related manner and normalized the other parameters studied except that plasma FSH remained slightly elevated. In contrast, when substitutive therapy was started 8 days after castration or later, the 7-day treatment with sex steroids reduced plasma gonadotropins, but pit GnRH-R remained elevated, and hypothalamic GnRH content remained depleted. These results indicate that the marked increase of gonadotropin secretion after castration is mediated at least in part, by an increase in the number of pit GnRH-R. Sex steroids were able to reverse all castration-induced endocrine changes in acutely castrated rats, but in long term castrated animals their action at higher centers to normalize hypothalamic GnRH content, and indirectly, to reduce pit GnRH-R content, was either delayed or ineffective. Thus, the rapid feedback action of sex steroids in long term castrated rats may be predominantly exerted at the pituitary level. PMID- 6274627 TI - Ethanol-induced release of adenosine 3',5'-monophosphate into the medium by rat testis in vitro. PMID- 6274628 TI - In vivo adrenocorticotropin-releasing activity of neurohypophyseal hormones and their analogs. AB - The ability of 21 neurohypophyseal hormones and related synthetic peptides to raise plasma ACTH or corticosterone concentrations was studied in female rats anesthetized with chlorpromazine, morphine, and pentobarbital. Corticotropin releasing factor (CRF) activity was significantly correlated with pressor but not with antidiuretic or oxytocic activity. However, peptides with little or no pressor but very potent antidiuretic activity had weak CRF activity if given in a dose greater than 4 antidiuretic units/100 g BW; the dose-response slopes of these analogs were significantly flatter than that of arginine vasopressin. Pretreatment with antagonists with antipressor but not antiantidiuretic activity consistently reduced the CRF activity of the analogs with potent pressor activity. We conclude that the CRF activity of the neurohypophyseal hormones is primarily related to pressor activity. PMID- 6274629 TI - Phosphatidylcholine synthesis and glycogen depletion in fetal mouse lung: developmental changes and the effects of dexamethasone. AB - We measured the rate of choline incorporation into phosphatidylcholine in lung slices; the glucogen content of the lung; and the activities of pulmonary cholinephosphate cytidylyltransferase, cholinephosphotransferase and phosphatidate phosphatase in the mouse during late fetal and early postnatal development. We also examined the effect of maternal dexamethasone administration on these parameters of fetal lung maturation. There was a development increase in the rate of choline incorporation between 17 days gestation (term is 19 days) and the immediate newborn period. There was also a developmental decrease in the glycogen content of the lung but this did not occur until 18 days. There was a developmental increase in the activities of cholinephosphate cytidylytransferase and phosphatidate phosphatase but little change in the activity of cholinephosphotransferase. Dexamethasone doubled the rate of choline incorporation into phosphatidylcholine at 17 and 18 days gestation. It decreased the glycogen content of the fetal lung by 74% at 18 and 19 days, but had no effect at 16 and 17 days. Dexamethasone increased the activity of pulmonary cholinephosphate cytidylyltransferase by 37% and that of cholinephosphotransferase by 27% at 17 days. It increased the activity of phosphatidate phosphatase by 25% at 16 days and by 32% at 19 days. These data show that the normal development profile of these parameters of fetal lung maturation in the mouse, as well as the effects of glucocorticoids thereon, are generally similar to those in the rabbit and rat. However, stimulation of cholinephosphotransferase by glucocorticoids has not been generally observed in other species. Furthermore, since the changes in the rate of choline incorporation precede those in glycogen depletion, the data suggest that the relationship between phospholipid synthesis and glycogen degradation is not simply that of precursor to product. PMID- 6274630 TI - In vivo removal of a few heavy metals in certain tissues of the fish Notopterus notopterus. PMID- 6274631 TI - Effect of acute treatment with cadmium on ethanol anesthesia, body temperature, and synaptosomal Na+-K+-ATPase of rat brain. PMID- 6274632 TI - Effects of silica on human lung fibroblasts: survival data analysis of time-lapse cinematography data. PMID- 6274633 TI - Quartz hemolysis as related to its surface functionalities. PMID- 6274634 TI - A network model for the organization of type IV collagen molecules in basement membranes. AB - Type IV collagen was solubilized from a tumor basement membrane either by acid extraction or by limited digestion with pepsin. The two forms were similar in composition and the size of the constituent chains but differed when examined by electron microscopy and in the fragment pattern produced by bacterial collagenase. The acid-soluble form showed after rotary shadowing strands mainly of a length of 320 nm which terminated in a globule, or two strands connected by a similar globule. The globule was identified as a non-collagenous domain (NC1) which under dissociating conditions could be separated into two peptides showing a monomer-dimer relationship. Higher aggregates of NC1 were visualized under non dissociating conditions. Some of the acid-extracted molecules have retained the previously 7-S collagen domain. The pepsin-solubilized form lacked domain NC1 and consisted mainly of four triple-helical strands (length 356 nm) joined together at the 7-S domain (length 30 nm). Common to both forms of type IV collagen was a small collagenase-resistant domain NC2 which was composed of collagenous and non collagenous elements and located between the 7-S domain and the major triple helix. These data indicate that the collagenous matrix of basement membranes consists of a regular network of type IV collagen molecules which is generated by two different interacting sites located at opposite ends of each molecule. The 7 S collagen domain connects four molecules while the NC1 domain connects two molecules. The maximal distance between identical cross-linking sites (7-S or NC1) was estimated to be about 800 nm comprising the length of two molecules. PMID- 6274635 TI - Binding of human carbonic anhydrase to human hemoglobin. AB - The ability of human carbonic anhydrases to interact with human CO-hemoglobin have been studied with the counter-current distribution technique in aqueous/aqueous biphasic systems. The experimental results show that human carbonic anhydrase II interacts with human CO-hemoglobin whereas human carbonic anhydrase I does not. THe interaction between CO-hemoglobin and carbonic anhydrase II was quantified using the theoretical model developed previously for one-to-one interacting systems. [Backman, L. and Shanbhag, V.P. (1979) J. Chromatogr. 171, 1-13]. The apparent association constant was estimated to be 4.1 x 10(5) l mol-1 at pH 8.0 and 21 degrees C for the association of carbonic anhydrase II and CO-hemoglobin. PMID- 6274636 TI - Platelet and leucocyte calmodulins: isolation and characterisation. AB - The calcium-dependent regulatory proteins, calmodulins, have been isolated from human blood platelets and guinea pig peritoneal polymorphonuclear leucocytes using the urea methanol procedure of Grand et al. [Biochem. J. 177, 521-529 (1978)]. The calmodulins were purified to homogeneity as indicated by polyacrylamide gel electrophoresis and both proteins comigrated with bovine brain calmodulin with mobilities corresponding to molecular weights of 16 000-17 000. The yield of calmodulin from platelets was higher on a wet weight basis than the yield from leucocytes but the former compared favourably with yields reported for brain and other tissues. Both calmodulin preparations significantly stimulated brain cyclic nucleotide phosphodiesterase, erythrocyte ghost Ca2+ ATPase and platelet phosphorylase kinase activities at the microgram level. Stimulation of Lubrol-solubilised brain adenylate cyclase was only marginally significant with platelet calmodulin and rarely demonstrable with the leucocyte preparations. Although biological activities of both proteins were retained during storage at 20 degrees C, higher-molecular-weight aggregates slowly formed which could not be dissociated during dodecylsulphate/mercaptoethanol denaturation. PMID- 6274638 TI - Electron transport to nitrogenase in Klebsiella pneumoniae: purification and properties of the nifJ protein. AB - In Klebsiella pneumoniae, the physiological electron flow to nitrogenase involves specifically, in addition to nitrogenase reductase, the products of the nifF and nifJ genes. The J protein was purified to homogeneity and was found to be an iron sulfur protein devoid of molybdenum. In its native state, the J protein is a dimer of Mr about 245 000, made up of two subunits of the same molecular weight. It contains about 30 mol iron and 24 mol labile sulfur/mol protein. The addition of J protein to crude extracts of a nifJ mutant reestablishes pyruvate-supported acetylene-reducing activity. This activity is further enhanced by addition of pure nitrogenase (Kp1). Based on its physical properties, the J protein is probably an oxidoreductase whose physiological role might be to transfer electrons from a metabolic donor to the F protein. In addition, another protein whose activity is also dependent on the nifJ gene seems to be required for the formation of a fully active Kp1. PMID- 6274637 TI - Electron transfer between azurin from Alcaligenes faecalis and cytochrome c551 from Pseudomonas aeruginosa. AB - The electron transfer equilibrium and kinetics between azurin from Alcaligenes faecalis and cytochrome c551 from Pseudomonas aeruginosa have been studied. The equilibrium constant K = ([Cyt(III)] . [Az(I)])/([Cyt(II)] . [Az(II))]) = 0.5 at 25 degrees C is about seven times smaller than that observed between the cytochrome c551 and the titrations confirmed a 43-mV difference between the mid point potentials of +266 mV and +309 mV for the Alcaligenes and Pseudomonas azurins respectively. The kinetics of the reaction between Alcaligenes azurin and Pseudomonas cytochrome c551 were investigated by the temperature-jump chemical relaxation method. Only a single relaxation mode was observed throughout the range of concentrations and temperatures examined. Thus, the slow relaxation time observed in the reaction between P. aeruginosa azurin and cytochrome c551 is not observed with the Alcaligenes azurin. The simplest mechanism that can therefore be ascribed to the investigated system is: [formula: see text]. This scheme is similar to that proposed earlier for the reaction between P. aeruginosa azurin and cytochrome c551 but does not involve the conformational transition proposed for azurin. The specific rates for the electron transfer are still fast: 1.8 x 10(6) M-1 . s-1 and 3.0 x 10(6) M-1 . s-1 respectively at 25 degrees C. PMID- 6274639 TI - Beckwith-Wiedemann (exomphalos-macroglossia-gigantism--EMG) syndrome and malignant lymphoma. PMID- 6274640 TI - Specific binding of prolactin in human mammary tumors. PMID- 6274641 TI - The changing role of endocrine therapy in the management of advanced breast cancer. PMID- 6274642 TI - Autoradiographic visualization of rat brain adenosine receptors using N6 cyclohexyl [3H]adenosine. PMID- 6274643 TI - Ontogeny of opioid pharmacology and receptors: high and low affinity site differences. AB - The development of the high affinity binding of a variety of opiates and enkephalins is distinct from low affinity binding. During the first 2 weeks after birth, low affinity binding in both brain and spinal cord remains relatively constant while high affinity binding increases up to 3-fold. Differences in the development of analgesic and respiratory sensitivity to opiates are also found. Whereas morphine, beta-endorphin and D-Ala2-Met5-enkephalin-amide depress respiratory rates in 2-day old rats at doses equal to or lower than those active in 14-day old rats, morphine's analgesic ED50 is 40-fold greater in 2-day old than in 14-day old rats. Similarly, beta-endorphin and D-Ala2-Met5 enkephalinamide are analgesic in 14-day old rats but not 7-day old rats. The small effect of spinal transections on morphine analgesia in 14-day old rats suggests that the change in analgesic sensitivity is at a segmental spinal level and not a result of descending pathways. These results suggest an interesting correlation between high affinity binding and analgesia and between low affinity binding and respiratory effects. PMID- 6274644 TI - Comparative actions of substances affecting cyclic AMP metabolism on the isometric contractions of fast and slow skeletal muscle during single-pulse and subtetanic stimulation. AB - Increasing concentrations of isoprenaline (0.5-4 muM) produced a dose-dependent increase in both TD and dT/dtmax during direct single-pulse stimulation of hemidiaphragm of the rat. The same drug during the same type of stimulation produced an insignificant change in these parameters of the isometric contraction of the isolated guinea-pig soleus muscle. On the contrary, isoprenaline produced a dose-dependent decrease of the isometric contraction during subtetanic stimulation of the soleus muscle. Contrary to the results obtained on hemidiaphragm, there was no interaction between halothane and aminophylline on the soleus muscle. In the soleus muscle, aminophylline (0.3-3.2 mM) produced a dose-dependent increase in TD and dT/dtmax during single-pulse stimulation, whereas isoprenaline failed to do so under the same experimental conditions, in spite of the fact that both substances are activators of cyclic AMP system. The beta2-selective adrenoceptor agonist terbutaline acted in the same way as isoprenaline. During subtetanic stimulation aminophylline (0.3-3.2 mM) produced a dose-dependent decrease of both parameters of the isometric contraction of hemidiaphragm, which is opposite to the results obtained during single-pulse stimulation. It is concluded that various types of electrical stimulation can produce different responses in slow and fast-contracting muscles, depending on the fundamental biochemical differences of two types of muscle, but some of these responses are the same irrespective of the method of muscle activation. PMID- 6274645 TI - Opiate receptor-binding activity of [D-Thr2,Thz5]-and [D-Met2,Thz5] enkephalinamides. AB - The binding characteristics of two highly potent analgesic enkephalin analogs, [D Thr2, Thz5]-and [D-Met2, Thz5]-enkephalinamide, to the rat brain membrane preparation have been investigated using tritiated Leu-enkephalin, human beta endorphin or dihydromorphine as the primary ligand. Concentrations for 50% inhibiting activity of the enkephalin analogs are much lower in displacing tritiated dihydromorphine in comparison with that for the other two tritiated ligands. In addition, a correlation between analgesic potency and capacity to displace tritiated dihydromorphine is observed. These data may explain the high antinociceptive activity of the enkephalin analogs and support the hypothesis that the mu-receptor is involved with analgesia. PMID- 6274646 TI - High-K+-induced inhibition of cAMP accumulation in smooth muscle. AB - This study examines a possibility that high-K+ depolarization induced inhibition of cAMP accumulation by beta-adrenoceptor stimulation (observed in our previous studies) is mediated by depolarization induced release of acetylcholine. Rat uterine strips were depolarized by high-K+ solution with or without atropine. Isoproterenol produced similar degrees of relaxation and increases in cAMP levels in depolarized smooth muscle with or without atropine. It is suggested that the release, if any, of acetylcholine from smooth muscle depolarization is not the primary cause of the observed inhibition of cAMP accumulation. PMID- 6274647 TI - Influence of naloxone and methysergide on the analgesic effect of clomipramine in rats. AB - The effects of the tricyclic antidepressant clomipramine were studied in two analgesic tests in rats: (1) vocalization threshold response; and (2) scored behavioral response to electric shock to the tail. Clomipramine (20-50 mg/kg i.p.) produced analgesia, decreasing behavioral response scores and increasing vocalization threshold. Morphine also reduced the response scores in the second test. Naloxone (0.8 mg/kg i.p) or methysergide (20 mg/kg i.p.) (no effect when given alone) abolished the analgesic effect of clomipramine as evaluated by vocalization threshold response. Naloxone alone (0.6 or 2 mg/kg i.p.) increased the behavioral response at 20 and 30 V but did not modify the score at 40 V. Naloxone reduced the analgesic effect of clomipramine or morphine in the behavioral test. These results suggest that the analgesic effect of clomipramine could involve both serotonergic and endorphin central systems. PMID- 6274648 TI - alpha 1-Adrenoceptor-mediated responses in the lateral geniculate nucleus are enhanced by chronic antidepressant treatment. AB - Responses of single dorsal lateral geniculate neurons to iontophoretic noradrenaline, serotonin, and carbachol were studied following acute or chronic administration of tricyclic antidepressants. Long-term (15-20 day) treatment of rats with a variety of clinically effective tricyclics, including the atypical iprindole, significantly enhanced alpha 1-adrenoceptor mediated activations induced by noradrenaline. This change appears to require chronic treatment, since acute and short term (6-8 day) tricyclic regimens fail to consistently affect noradrenaline's action. Long term antidepressant treatment was also effective in enhancing geniculate neuron sensitivity to serotonin, in accord with previous studies, but failed to modify responses to the cholinergic agonist carbachol. It is suggested that the modulation of noradrenaline and serotonin receptor activity may represent a slowly developing action of tricyclic antidepressants which correlates with their delayed clinical effects. PMID- 6274649 TI - Adrenal steroid-induced changes in beta-adrenergic receptor binding in rat hippocampus. AB - Bilateral 6-hydroxydopamine (6-OHDA) lesions of the dorsal noradrenergic bundle have been shown to increase [3H]dihydroalprenolol (DHA) binding in the rat hippocampus. The effect of adrenalectomy on this increase in receptor binding was examined. Groups of rats received stereotaxically placed 6-OHDA injections aimed at the dorsal bundle (DB) or vehicle injections (control). Half of each group subsequently received bilateral adrenalectomies (ADX) and the other half received sham operations, yielding four groups: control, DB, ADX, and DB + ADX. The DB lesions produced a 41% increase in maximum binding in close agreement with a previous report. No statistically significant change in [3H]DHA receptor binding was observed following adrenalectomy treatment alone. By contrast, the DB + ADX group showed a significant increase in maximum [3H]DHA receptor binding compared to the DB group. This adrenalectomy-induced increase was reversed after 1 week of corticosterone treatment (1.0 mg/kg/12 h, s.c.). These results suggest an interaction between noradrenergic mechanisms and corticosterone in the hippocampus. PMID- 6274650 TI - Atypical characteristics of frog and chick erythrocyte beta-adrenoceptors. AB - The characteristics of beta-adrenoceptors on frog and chick erythrocytes have been compared to those present on rat erythrocyte, lung and cerebral cortical membranes. The receptors have been examined using the specific ligand [3H]dihydroalprenolol ([3H]DHA) with conventional filtration techniques. The equilibrium dissociation constant (KD) of [3H]DHA and the affinities of the non selective beta-adrenoceptor antagonists propranolol, timolol and alprenolol, were very similar in all preparations examined. The relative order of potency of the catecholamines, isoprenaline greater than adrenaline greater than noradrenaline in rat lung, rat erythrocyte and frog erythrocyte; isoprenaline greater than noradrenaline greater than or equal to adrenaline in chick erythrocyte and rat cortex, suggest an overall beta 2-classification for the former and beta 1 classification of the latter tissues. However, the use of highly selective beta 1 or beta 2-agents and computer-assisted iterative curve fitting revealed that whereas rat cortex and lung possess both receptor subtypes (lung 80% beta 2, 20% beta 1: cortex 65% beta 1, 35% beta 2) only beta 2-sites were observed in rat erythrocytes. On the other hand, chick and frog erythrocytes possess a homogeneous population of receptors that do not strictly correspond either to beta 1- or beta 2-adrenoceptors. The atypical nature of these non-mammalian erythrocyte beta-adrenoceptors is discussed in relation to their previous extensive use as model beta-adrenoceptor systems. PMID- 6274651 TI - Effect of N-methylpyridinium-2-aldoxime methane sulphonate (P2S) on rat intestinal, (Na-K)ATPase and adenyl cyclase activities. AB - The effect of N-methylpyridinium-2-aldoxime methane sulphonate (P2S), a drug recommended for prophylactic and therapeutic purposes in organophosphate poisoning, on intestinal (Na-K) ATPase and adenyl cyclase activities, was tested in rats. Intestinal (Na-K) ATPase activity was determined 5 h after intragastric administration of either 0.15 M NaCl or P2S 200 mg/kg body weight. P2S decreased significantly jejunal and colonic (Na-K)ATPase activity, 17.1 +/- 4.8 (S.E.) and 13.5 +/- 3.0, as compared to that in saline-treated rats, 41.5 +/- 3.0 (S.E.) and 25.4 +/- 1.2 mumol Pi/mg protein per h, respectively. Pretreatment with methyl prednisolone did not prevent the decrease in enzyme activity induced by P2S. Mucosal PGE2 and cAMP contents, adenyl cyclase and phosphodiesterase activities, were similar in P2S and saline-treated rats. It is thus suggested that P2S induced inhibition of intestinal (Na-K)ATPase activity might be among the mechanisms contributing to looseness of the stool frequently observed following P2S administration. PMID- 6274652 TI - Comparative actions of mexiletine on sodium channels in nerve, skeletal and cardiac muscle. AB - Mexiletine's actions on voltage-clamped sodium channels of frog myelinated nerve and skeletal muscle are described. Mexiletine blocks half the sodium channels (infrequent depolarizations) of single myelinated nerves at a 83 micro M concentration while only 26 micro M is required to do the same in skeletal muscle preparations where similar vaseline-gap techniques are utilized. Mexiletine's potency for block of sodium current in nerve is clearly related to its lipid distribution characteristics given proper consideration of the drug class to which it belongs. Hyperpolarizing prepulses, which are typically used to remove normal sodium inactivation, appear to reduce drug blocking potency suggesting that noninactive channels have a considerably lower affinity for the drug than do inactive channels. Direct evidence supporting selective drug block of inactive channels is also given. In addition the effects of this drug on sodium channels of guinea pig papillary muscle have been studied using measurements of maximum upstroke velocity of intracellularly recorded action potentials. In these myocardial studies 5 to 20 micro M mexiletine depressed upstroke velocity of papillary muscle action potentials in a frequency-dependent fashion. No basal (nonfrequency-dependent) block was observed in heart at these therapeutic concentrations of mexiletine. comparisons are made between skeletal and cardiac muscle effects of mexiletine, especially relating to the important role played by sodium channel inactivation. PMID- 6274653 TI - The effect of trypsin of rate, force and cyclic AMP in guinea pig atria. AB - Trypsin (10(-6)M produced a positive inotropic and chronotropic effect in left and right atria respectively and an increase in cyclic AMP. The effects were blocked by aprotinine while propranolol, phentolamine, promethazine and cimetidine and reserpine pretreatment did not alter trypsin activity. Trypsin effects were potentiated by RO 20,1724, a phosphodiesterase inhibitor. The cardiac effects of trypsin may be due to increase in cyclic AMP and are in agreement with previous work indicating that trypsin can activate cardiac adenylate cyclase. PMID- 6274654 TI - Naloxone inhibits and morphine potentiates the adrenal steroidogenic response to ACTH. AB - The administration of morphine to hypophysectomized rats potentiated the steroidogenic response of the adrenal cortex to exogenous adrenocorticotrophic hormone (ACTH) in a dose-dependent fashion. Conversely, the opiate antagonist naloxone inhibited the adrenal response to ACTH. Naloxone pretreatment also antagonized the potentiating effect of morphine on ACTH-induced steroidogenesis in a dose-dependent manner. Neither morphine nor naloxone, administered to hypophysectomized rats, had any direct effect on adrenal steroidogenesis. These adrenal actions were stereospecific since neither the (+)-stereoisomer of morphine, nor that or naloxone, had any effect on the adrenal response to ACTH. The administration of human beta-endorphin to hypophysectomized rats had no effect on the adrenal corticosterone concentration nor did it alter the response of the adrenal gland to ACTH. These results indicate that morphine can potentiate the action of ACTH on the adrenal by a direct, stereospecific, dose-dependent mechanism that is prevented by naloxone pretreatment and which may involve competition for ACTH receptors on the corticosterone-secreting cells of the adrenal cortex. PMID- 6274655 TI - Effect of norepinephrine uptake blocker on beta-adrenergic receptors of the rat cerebral cortex. PMID- 6274656 TI - Functional characterization of postjunctional alpha-adrenoceptors in rat aorta. PMID- 6274657 TI - Effect of naloxone on the cardiovascular and sympathetic response to hypovolemic hypotension in the rat. PMID- 6274658 TI - Alpha-noradrenaline modulation of D,L-allylgycine seizures. AB - The preferential alpha 2-noradrenergic agonist clonidine dose-relatedly increased the onset of seizures and mortality times, and decreased severity in rats treated with D,L-allylglycine. These effects were reduced by a dose (2.5 mg/kg) of the preferential alpha 2-antagonist yohimbine, which was itself inactive on the allylglycine seizures. Yohimbine 10 mg/kg alone decreased onset and mortality times. The preferential alpha 1-antagonist prazosin had a slight anticonvulsant effect. These results suggest that the reduction of NA release at alpha 2 noradrenergic receptors or antagonism of noradrenaline effects at alpha 1 receptors exerts seizure suppressant effects. PMID- 6274659 TI - Reversal of the antiaversive and anticonvulsant actions of diazepam, but not of progabide, by a selective antagonist of benzodiazepine receptors. PMID- 6274660 TI - Rapid changes in cerebellar adenosine receptors following experimental seizures. PMID- 6274661 TI - Psychotomimetic opioid binding: specific binding of [3H]SKF-10047 to etorphine inaccessible sites in guinea-pig brain. PMID- 6274662 TI - Effect of acute and chronic pentylenetetrazol treatment on benzodiazepine and cholinergic receptor binding in rat brain. AB - The binding of [3H] diazepam and [3H] flunitrazepam in rat cerebral cortex was not altered by either acute or chronic administration of pentylenetetrazol except in rats made to convulse 30 min before sacrifice. Rats treated for up to 6 months with doses of pentylenetetrazol which are below seizure threshold in naive rats, became increasingly sensitive to the CNS stimulant effect of pentylenetetrazol as demonstrated by the development of myoclonus and convulsions during treatment periods. These effects were not correlated with any changes in benzodiazepine binding in cerebral cortex or cerebellum and [3H] quinuclidinyl benzilate binding in cerebral cortex. Acute convulsant doses of pentylenetetrazol increased benzodiazepine binding in cerebral cortex, but only in those rats which actively convulsed. Benzodiazepine and cholinergic receptors of the cortex, and benzodiazepine receptors of the cerebellum, therefore, do not appear to change with either the acute or chronic subconvulsive administration of pentylenetetrazol. PMID- 6274663 TI - Prostacyclin binding to guinea pig pulmonary receptors. AB - [3H] Prostacyclin bound to membranes of guinea pig lung. Specific binding was saturable, and the results revealed two receptor classes (Kd =16 nM and 258 nM). The binding capacity of the high affinity site was 105 fmol [3H]prostacyclin per mg of membrane protein, and that of the low affinity site was 1257 fmol per mg membrane protein. A comparison of selected prostaglandins as inhibitors of [3H] prostacyclin binding revealed some of the structural requriements of the ligand for occupation of the high affinity receptor. PMID- 6274664 TI - Affinity of thiocolchicoside and thiocolchicoside analogues for the postsynaptic GABA receptor site. PMID- 6274665 TI - Endogenous GABA determines the characteristics of [3H]GABA-binding. AB - Since the identification of sodium-independent GABA-binding sites in membrane preparations from the mammalian central nervous system numerous publications have dealt with the effects of putative endogenous inhibitors of GABA-binding. However, controversy has arisen over the existence and identity of such inhibitors and the possible artifactural role of residual endogenous GABA in the interpretation of GABA-binding data. Using direct determination of GABA concentrations we have demonstrated that it is extremely difficult to remove endogenous GABA from rat brain membrane preparations and have shown that, even in well-washed preparations, there is a source of GABA production which compounds this problem. When care has been taken to ensure that GABA concentrations are minimal, analysis of binding data reveals the presence of two classes of GABA binding sites with dissociation constants of the order of 9 nM and 318 nM and maximum binding capacities of 0.63 and 1.65 pmol/mg protein respectively (i.e., 63 and 165 pmol/g original wet tissue). A theoretical analysis of the effects of endogenous GABA demonstrates how failure to take into account even very low concentrations of GABA leads to misinterpretation of the results. PMID- 6274666 TI - Mammalian beta-adrenoceptors: concomitant biospecific elution with protein kinase activity from Sepharose-alprenolol. AB - The mammalian beta-adrenoceptors were solubilized from crude guinea pig lung membranes with the plant glycoside digitonin. The solubilized receptors were absorbed by affinity gels with (+/-)alprenolol as ligand and could be eluted biospecifically in good yield with either agonists or antagonists. A more than 100-fold purification can be achieved in a one-step procedure. Antagonist-eluted beta-adrenoceptors have affinities for antagonists similar to those of particulate or solubilized receptors. Their binding constants for agonists, however, are comparable to those of the particulate receptor. A cAMP-independent protein kinase activity was eluting concomitantly with the beta-adrenoceptors. The protein kinase phosphorylated endogenous substrates with 50 000 and greater than 150 000 subunit molecular weight. PMID- 6274667 TI - Calcium ions, morphine tolerance and noradrenergic transmission in the mouse vas deferens. AB - The response of the isolated vas deferens of the mouse to electrical stimulation is inhibited by morphine and levorphanol via an opiate receptor, the inhibition decreasing with increasing stimulation frequency (0.2-16 Hz). Tolerance to the locomotor stimulant effect of morphine was induced over 48 h using a slow release preparation. Vasa from mice similarly treated with the slow release preparation showed a shift to the right of the morphine and levorphanol twitch inhibition curves. The reduction in the fractional release of [3H]noradrenaline by morphine and levorphanol was less in vasa from morphine-pretreated mice. Altering the Krebs solution by reducing the Ca2+ or Na+ or adding Mg2+ increased the effect of opiate agonists in vasa from naive and morphine-tolerant mice. Therefore, tolerance to morphine has not changed the ability of these ions to modulate opiate responses. PMID- 6274668 TI - Does ethanol stimulate brain opiate receptors? Studies on receptor binding and naloxone inhibition of ethanol-induced effects. AB - The ability of ethanol to stimulate opiate receptors was investigated. Administration of ethanol 2-3.5 g/kg in rats induced analgesia as tested by the tail flick method, reduced body temperature, impaired sensorimotor performance and induced "sleep". Naloxone HCI (0.5-10 mg/kg) did not significantly attenuate these ethanol-induced behavioural changes. [3H]Etorphine binding to opiate receptors in membrane preparations from rat and mouse brains was unchanged by ethanol concentrations of 0.05-3% (v/v). It was concluded that the ethanol effects studied were not mediated by naloxone-sensitive opiate receptors. PMID- 6274669 TI - Suppressive action of cholecystokinin octapeptide on the behavioral effects of L DOPA in the rat. AB - Intracerebroventricular (i.c.v.) administration of cholecystokinin octapeptide (CCK-8) appeared to suppress the effects of L-DOPA and antagonize the actions of thyrotropin-releasing hormone (TRH). This was not only proved by the behavioral rating, but also by locomotor activity counting. thus, the sedative action of CCK 8 and the antagonizing effect of this peptide on the central actions of TRH were clearly demonstrated. PMID- 6274670 TI - Presynaptic effects of 4-aminopyridine and streptomycin on the neuromuscular junction. AB - Studies were done to assess the effects of 4-aminopyridine (4AP) and streptomycin (SM) on transmitter release parameters and extracellularly recorded presynaptic action potential. The application of 5 micrometer 4AP resulted in a marked increase in the mean quantal content (m1) associated with an increase in the total number of the store of available quanta (n) but had no effect on the probability of release (p) and the fractional release (P). Focal recording showed that 50 micrometer 4AP modified the shape of the presynaptic action potential from a triphasic configuration to a diphasic one. In contrast, prolongation of the muscle action potential was found only at higher concentrations (greater than 1 mM) of of 4AP. Thus, the increase in n with 4 AP was associated with prolongation of the presynaptic action potential evoked by blocking the K current. SM (172 micrometer mM) had no effect on p and P. Reduction of n by SM was completely reversed by 4AP. As the presynaptic action potential change induced by 4AP was not antagonised by SM, it may be that the decrease of n by SM followed a modification of the voltage-dependent Ca channel. PMID- 6274672 TI - Are there subtypes (isoreceptors) of multiple opiate receptors in the mouse vas deferens? AB - The hypothesis of heterogeneity of opiate receptors was tested with the electrically stimulated mouse vas deferens. The experimental approach was based on the phenomenon of tolerance development of selective opiate receptors. It was found that opioids classified as mu-receptor agonists, e.g. sufentanyl and normorphine, do not necessarily exhibit cross-tolerance to each other. The same holds true for a variety of presumed kappa-receptor agonists. On the other hand, the delta-receptor agonists under investigation displayed complete cross tolerance. These findings give rise to an interpretation which may postulate the existence of subtypes (isoreceptors) of the already known opiate receptors. PMID- 6274671 TI - Excitability ot beta-endorphin, morphine and catecholamines of the vas deferens of the rat at different stages of development. AB - The effect of beta-endorphin and morphine on neuromuscular twitching of the vas deferens was examined in rats of 10-120 days of age. The maximal tension developed by the smooth muscle was markedly dependent on the age of the rat and on the kind of stimulus applied. The potency of beta-endorphin to inhibit the electrically induced twitching or of catecholamines to induce muscle contraction was not significantly altered by development of the animal. The excitatory action of morphine showed significant variations in activity reaching maximal sensitivity at about two months of age. Age or the onset of puberty do not significantly modify the sensitivity of the vas deferens to beta-endorphin, the peptide may play the role of modulating adrenergic transmission. PMID- 6274673 TI - Effects of corticotrophin (ACTH) on recovery of sensorimotor function in the rat: structure-activity study. AB - The recovery of sensorimotor function in rats was studied using a foot-flick response test after crushing the sciatic nerve. Every other day, the animals received a subcutaneous injection of ACTH1-24 or vehicle, immediately after the 'foot-flick' test. Rats treated with ACTH1-24 showed a faster recovery of sensorimotor function as compared to vehicle-treated rats. This beneficial effect was dose-dependent. In contrast, no effect on toe-spreading could be detected. To investigate what part of the peptide molecule would contain the active site and to exclude steroid mediation, smaller sequences of ACTH1-24 devoid of corticotrophic activity were tested. Treatment with ACTH1-16NH2, ACTH4-10 or [Met(O2)4,D-Lys8,Phe9]ACTH4-9 (Org. 2766) mimicked the effect of ACTH1-24, whereas treatment with ACTH11-24 did not effect the return of sensorimotor function. The stimulatory effects of corticotrophin1-24 and congeners on recovery of sensorimotor function are discussed in terms of a direct effect of these peptides on nervous tissue, probably through enhanced brain and spinal cord protein synthesis. PMID- 6274674 TI - Postsynaptic spinal alpha-adrenoceptors mediate effects of intrathecal clonidine. AB - The cardiovascular effects of intrathecal administration of clonidine were examined after depletion of spinal cord noradrenaline or 5-hydroxytryptamine by pretreatment with 6-hydroxydopamine or 5,6-dihydroxytryptamine. Despite marked reduction in spinal noradrenaline or 5-hydroxytryptamine levels, resting blood pressure and heart rate were unchanged and the clonidine-induced hypotension and bradycardia were unimpaired. The results suggest that intrathecal clonidine acts via inhibitory spinal alpha-adrenoceptors located postsynaptically on the preganglionic nerve cell bodies. PMID- 6274675 TI - [Antibody production in rats vaccinated with inactivated Sendai virus (author's transl)]. AB - Adult male and female rats were inoculated with tween 80-ethylether-formalin ultraviolet inactivated Sendai virus (Sv-V) and examined for the production of hemagglutination inhibition (HI) antibody. There were no significant differences in the antibody titers between males and females, and among the various routes of inoculation except for the intranasal which was not effective. The antibody became detectable 7 days after a single inoculation with 10(5) HAU of Sv-V. The antibody titer, which had its peak 21 days after the inoculation, persisted for 200 days and declined gradually thereafter. The HI antibody titers were correlated with inoculated Sv-V doses and a predominant booster reaction with the vaccine was observed. Maternal antibodies were detected in sucklings born to dams hyperimmunized with the vaccine. The titers were similar to those of the dams until 3 weeks after birth but declined rapidly after weaning at 4-week-old. The titers of fetuses and neonates before suckling were significantly lower than those of the sucklings. PMID- 6274676 TI - Prevalence of feline viral antibodies in random-source laboratory cats. AB - Over a period of 1973 to 1979, a serologic survey of virus infections was conducted on feline sera collected in four universities which located in different prefectures; Obihiro, Saitama, Kanagawa and Tokyo. A significant hemagglutination-inhibition (HI) antibody titer of 1 : 8 or higher to feline panleukopenia virus (FPLV) was detected in 130 (58%) of the 226 sera used. No remarkable difference in the HI antibody prevalence in cats to FPLV was recognized by years or localities. Of a total of 188 cats tested, 99 (53%) presented positive serum neutralizing (SN) antibody titers to the No. 1 strain of feline calicivirus (FCV). Especially in Kanagawa, 17 (77%) of the 22 cats had positive SN titers. However, only 42 (22%) of the 188 sera showed positive SN titers to the Kyoritsu strain of FCV. Such lower positivity in the cats was observed with 13% in the SN test to human reovirus type 3 (Reo-3). The incidence of positive SN antibodies to feline rhinotracheitis virus (FRV) also remained in low values of 20 to 27% with the exception of high percentage of 86 in Tokyo. The dissemination of FPLV, FRV, FCV and Reo-3 was briefly discussed in relation with the age distribution of viral antibodies in cats. PMID- 6274677 TI - Effects of Sendai virus infection on body weight, body temperature and some hematological and serobiochemical values of mice. AB - Body weight and body temperature of JCL: ICR mice infected with Sendai virus decreased depending on the viral dose. In severely infected mice, there were significantly higher values of hemoglobin, hematocrit, red and white blood cell counts, and leucin aminopeptidase (LAP), and lower values of reticulocytes and lactic dehydrogenase (LDH). The other values tested were comparable to those of the non-infected mice. PMID- 6274678 TI - A comparison between the enzymes in the cornea of the vitamin-A deficient rat and those of rat leucocytes. PMID- 6274679 TI - Desensitization of the beta-adrenergic receptor-adenylate cyclase complex in rabbit iris-ciliary body induced by topical epinephrine. PMID- 6274680 TI - The brachymorphic mutation of mice and altered developmental patterns of limb bud 3':5' cyclic adenosine monophosphate. AB - Concentrations of cyclic AMP (cAMP) were determined in paired fore and hind limbs from day 12-16 of development in murine fetuses homozygous for the brachymorphic (bm) mutation and normal controls. A developmental rise in cAMP occurred 1 day earlier in bm/bm than in +/+ hind limbs and cAMP was higher in day-13 bm/bm than in +/+ fore limbs. Since cAMP is well documented to stimulate chondrogenic differentiation, premature cartilage determination secondary to altered levels of cAMP could play a role in bm/bm short-limbed dwarfism. PMID- 6274681 TI - Effect of cyclonicate, a new hypolipemic drug, on lipolysis in vitro. AB - A new hypolipemic drug, cyclonicate (3,3,5-trimethyl-cyclohexyl nicotinate), was studied and its effects were compared with those of nicotinic acid. Experiments were carried out in vitro on rat adipose tissue on spontaneous lipolysis, and on lipolysis stimulated by noradrenaline, theophylline, dibutyryl cyclic AMP. Cyclonicate, like nicotinic acid, reduced the lipolytic stimulation of theophylline. Its effect was dose-dependent but 10 times lower (IC50 = 1 mM) than that of nicotinic acid. The inhibitory activity of both compounds was higher on theophylline-induced lipolysis than on noradrenaline-stimulated lipolysis. Neither one had any effect on dibutyryl cyclic AMP. In the absence of stimulating drugs, cyclonicate increased the FFA/glycerol ratio, more than nicotinic acid. Moreover, cyclonicate inhibited theophylline-induced FFA release much less than glycerol release, while under the same conditions, nicotinic acid inhibited both FFA and glycerol release. Thus cyclonicate would influence not only triglyceride hydrolysis, but also FFA utilization by adipose tissue. PMID- 6274682 TI - On the function of multiple subunits of cytochrome c oxidase from higher eukaryotes. PMID- 6274683 TI - Active site-directed inhibition of galactosidases by conduritol C epoxides (1,2 anhydro-epi- and neo-inositol). PMID- 6274684 TI - Is the acid phosphatase of Escherichia coli with pH optimum of 2.5 A polyphosphate depolymerase? PMID- 6274685 TI - Mr-values of mature subunits I and III of beef heart cytochrome c oxidase in relationship to nucleotide sequences of their genes. PMID- 6274686 TI - The production of oxygen-derived radicals by neutrophils from selenium-deficient cattle. PMID- 6274687 TI - Presence of fructose-6-phosphate,2-kinase in pancreatic islets. PMID- 6274688 TI - Epinephrine activation of phosphofructokinase in perfused rat heart. PMID- 6274689 TI - The NH2-terminal amino acid sequence of cellular retinoic-acid binding protein from rat testis. PMID- 6274690 TI - Cyclic AMP-dependent protein kinase of yeast. PMID- 6274691 TI - Isolation and structural organization of the human corticotropin--beta-lipotropin precursor gene. PMID- 6274692 TI - Multiplicity of mitochondrial proteinases in yeast. PMID- 6274693 TI - A re-evaluation of the role of fatty acids in the physiological regulation of the proton conductance of brown adipose tissue mitochondria. PMID- 6274694 TI - Topography of the iron-sulphur subunit in mitochondrial ubiquinol:cytochrome c reductase. PMID- 6274696 TI - Calmodulin and cyclic nucleotide-phosphodiesterase activities in rat mammary gland during the lactogenic cycle. PMID- 6274695 TI - Effect of Ca2+ on the inhibition of calcium-activated neutral protease by leupeptin, antipain and epoxysuccinate derivatives. PMID- 6274697 TI - Insulin inactivation of rat hepatocyte cyclic AMP-dependent protein kinase. PMID- 6274698 TI - Stimulation of human leukocyte degranulation by leukotriene B4 and its omega oxidized metabolites. PMID- 6274699 TI - Increases of prostacyclin and PGE2 levels after acute thyrotropin stimulation in cultured porcine thyroid cells. PMID- 6274700 TI - Conformational selection of syn-cAMP upon binding to the cAMP: receptor protein. PMID- 6274701 TI - Increases in protein A24 following treatment of Ehrlich ascites tumor cells with 1-methyl-1-nitrosourea and 1,3-bis(2-chloroethyl)-1-nitrosourea. PMID- 6274702 TI - Primary structure of the histone H3 and H4 genes and their flanking sequences in a minor histone gene cluster of Xenopus laevis. PMID- 6274703 TI - Ligand binding to the blue copper center of horse liver alcohol dehydrogenase. PMID- 6274704 TI - [Clinical aspects and diagnosis of hypertensive states in children and adolescents (I)]. PMID- 6274705 TI - [Effect of intrahypothalamic administration of gamma-aminobutyric acid on hypothalamic temperature and overall body-heat production]. PMID- 6274706 TI - [Effect of blockers of slow Na-Ca channels of frog myocardium on contractility and potassium contracture]. PMID- 6274707 TI - [The effects of the angiotensin converting enzyme inhibitor, captopril, on catecholamine concentrations in rat plasma, heart, brain and kidneys (author's transl)]. AB - Two studies were designed to examine the effect of the angiotensin converting enzyme inhibitor, captopril, on the sympathetic nervous system. In the first study, blood pressure (BP), heart rate (HR), and norepinephrine (NE) concentrations in the heart were measured in rats which had been given the agent orally in tap water (0.5 mg/ml) for 2, 9, 19, 29, and 58 days. BP and HR were measured using a tail-microphone to which a tachometer to record HR was connected in the unanesthetized and unrestricted condition. Heart NE was extracted with perchrolic acid and measured with the THI method on a high pressure liquid chromatography. The same study was also done in the control rats. The BP of the rats which had been given captopril for 9 days or more was significantly lower than in the control rats, while HR was not different between the two groups of rats. The ratio of heart weight/body weight was significantly lower in the captopril rats than in the control rats 58 days after the captopril administration. The ratio was significantly correlated with BP in these captopril and control rats (r = 0.59, p less than 0.01). In contrast to the control rats, the NE concentrations in the heart gradually increased in the captopril rats, thus being significantly higher in the latter than in the former after 29 and 58 days of captopril administration (p less than 0.01 for both observations). In addition, the lower the BP was, the higher the NE concentrations in the heart was in all the rats (r = 0.52, p less than 0.001). In the second study, BP, HR, and NE concentrations in plasma, heart, brain and the left kidney were measured in rats which had been on captopril for 2 and 29 days. Renal renin content (RRC) was also measured in the right kidney. In this study, BP and HR were recorded through a carotid catheter which had been inserted 4 hrs previously under light ether anesthesia. BP was significantly lower and NE concentrations in the heart was higher in the captopril rats than in the control rats after 29 days of captopril administration. There was a significant negative correlation between BP and NE concentrations in the heart in the captopril and the control rats (r = -0.88, p less than 0.001). No difference in HR was found between the 2 groups. NE concentrations in plasma, brain and kidney showed no significant differences between the captopril and the control rats in either of the stages of sacrifice. RRC was markedly reduced in the rats with 2 days of captopril, while it increased in the rats with 29 days of captopril. However, the RRC had no definite relationship with the NE concentrations in plasma or that in the kidney. The results show that chronic administration of captopril reduces the heart weight in normotensive rats. This effect points on the one hand to a decrease in "cardiac after-load" and on the other to changes of humoral factors such as plasma angiotensin II induced by captopril... PMID- 6274708 TI - [The correlation between serum angiotensin I-converting enzyme activity and the renin-angiotensin-aldosterone system in hypertensive patients (author's transl)]. AB - In order to investigate the role and origin of serum angiotensin I-converting enzyme activity (ACEA) in hypertension, the correlation between serum ACEA and the renin-angiotensin-aldosterone system was evaluated in hypertensive patients of whom 36 had essential hypertension, five had hypertension associated with chronic renal failure, three had renovascular hypertension, and one had primary aldosteronism. Serum ACEA was 17.0 +/- 3.3 U (mean +/- sd) in the normotensive control, 20.1 +/- 7.1 U in patients with essential hypertension, 11.6 +/- 3.4 U in patients with chronic renal failure, 31.7 +/- 1.1 U in patients with renovascular hypertension, and 9.7 U in primary aldosteronism in a recumbent state. There was a significant correlation between serum ACEA and plasma renin activity (PRA) (r = 0.615, p less than 0.001, n = 45) and plasma aldosterone concentration (PAC) (r = 0.599, p less than 0.001, n = 34) in a recumbent state. However, there was no significant correlation between serum ACEA and mean blood pressure. Serum ACEA elevated with furosemide and an upright posture significantly correlated with elevation in PRA (r = 0.369, p less than 0.05, n = 32) but did not significantly correlate with elevation in PAC. It is suggested, therefore, that the kidney is the suspected source of the plasma activity of the enzyme and that serum ACEA plays a possible role in the regulations of blood pressure and electrolyte metabolism modulating the renin-angiotensin-aldosterone system. PMID- 6274709 TI - [Serum and urine calcitonin levels in patients with medullary thyroid carcinoma and with non-thyroidal cancer, and their immunological size-heterogeneity (author's transl)]. PMID- 6274710 TI - [The study of serum dehydroepiandrosterone sulfate levels in normal children and children with pituitary dwarfism (author's transl)]. PMID- 6274711 TI - [Autoregulation of the gonadotropin receptors in rat testes after human chorionic gonadotropin administration (author's transl)]. PMID- 6274712 TI - [Studies on the measurement of vitamin D derivatives in human plasma. II. A fundamental study on the multiple assay for vitamin D derivatives (author's transl)]. PMID- 6274713 TI - [A case of pseudohypoparathyroidism with frequent normocalcemia (author's transl)]. PMID- 6274714 TI - Studies on interactions between plasminogen activator-containing fluids and avain and murine lymphocytes. PMID- 6274715 TI - Blood glucose and insulin levels in normal subjects following a meal with and without added sugar beet pulp. AB - The fibrous components of sugar beet pulp were investigated to determine whether they would reduce the post-prandial rise in blood glucose and plasma insulin levels when incorporated into a mixed meal. On two separate occasions six healthy volunteers were given either a control meal (providing 86 g carbohydrate) or an identical meal with the addition of 20 g sugar beet pulp (test meal). Blood glucose and plasma insulin levels were measured post-prandially for 3 hours. There was no significant difference between the mean blood glucose of plasma insulin curves at any time after the two meals. Since viscous types of dietary fibers are known to be effective in reducing post-prandial hyperglycaemia and insulinaemia, it would seem that either the physio-chemical nature of the fibre or the procedure employed to extract the sugar renders the particulate fibre inactive. PMID- 6274716 TI - Studies on the toxicity and binding kinetics of abrin in normal and Epstein Barr virus-transformed lymphocyte culture-I: experimental results - 1. AB - The effects of treatment with varying doses of abrin, a D-galactose binding lectin, on DNA and protein synthesis of normal and Epstein Barr virus-transformed lymphocytes have been investigated. Activation, stimulation and relative toxicity factor indices are studied as well as possible relationships between the DNA and protein synthesis rates, as measured by simultaneous 3H-TdR and 14C-leucine uptake. PMID- 6274717 TI - Studies on toxicity and binding kinetics of abrin in normal and Epstein Barr virus-transformed lymphocyte culture-I: experimental results - 2. AB - The effects of treatment with varying doses of abrin, a D-galactose binding lectin, on DNA and protein synthesis of normal and Epstein Barr virus-transformed lymphocytes have been investigated. Studies of activation and stimulation indices, as well as two new indices; the metabolic self and cross-coupling indices, lead to the prediction that there are three morphologically distinct subpopulations of EBV-transformed lymphocytes with different abrin binding site numbers. This conclusion is supported by SEM morphological differences. PMID- 6274718 TI - Studies on the toxicity and binding kinetics of abrin in normal and Epstein Barr virus-transformed lymphocyte culture-I: experimental results - 3. AB - The effects of treatment with varying doses of abrin, a D-galactose binding lectin, on DNA and protein synthesis of normal and Epstein Barr virus-transformed lymphocytes have been investigated. Using data on EBV-transformed lymphocyte cell density as a function of time and dose of abrin, one can demonstrate that the mean number of sites bound/EBV-lymphocyte needed to exert a biological influence upon the cell DNA synthesis lies between 59,264 and 370,000 sites/cell. Using a simple packing model, one can demonstrate that a theoretical estimate places the number of binding sites between 57,600 and 360,000 sites/cells. PMID- 6274719 TI - Allosteric and covalent control of glycogen synthase from leukocytes. PMID- 6274720 TI - Multiple forms of PRPP synthetase in human leukemic white blood cells seen with isoelectric focusing. PMID- 6274721 TI - Comparison between the effect of polycations and detergent on the specificity of cat liver microsomal glucose-6-phosphatase. PMID- 6274722 TI - NAD-glycohydrolase activity of staphylococcal entertoxin A (SEA). PMID- 6274723 TI - Contraception with LHRH agonists, a new physiological approach. PMID- 6274724 TI - Developmentally regulated transcription from Drosophila melanogaster chromosomal site 67B. PMID- 6274725 TI - Ketoconazole approved for systemic fungal infections. PMID- 6274726 TI - Strain difference in the expression of mammary tumor virus antigen in the male genital organs of mice during aging. AB - Mammary tumor virus (MTV) antigen expression was tested in the male genital organs (testis, epididymis, prostate, seminal vesicle and coagulating gland) of DD/Tbr, KF and DDD strains (dd (German) mouse group), GR strain (Swiss origin), SHN and SLN strains (different Swiss origin), C3H and A strains (American origin), BALB/c and C57BL strains (low mammary tumor strains), and DDf and SHNf strains (MTV eliminated strains). Microimmunodiffusion analysis showed that expression of MTV antigen is tissue extracts was found most frequently in the prostate, sometimes in the seminal vesicle and coagulating gland but rarely in the epididymis and not at all in the testis of mammary cancer strains. No expression was found in these organs from BALB/c and C57BL strains. The earliest expression was found in GR, SHN and SLN strains of Swiss origin. Expression was not lost in SHNf strain, indicating that it is due to genetically transmitted MTV, rather than a milk-factor. Male genital organs from two mammary cancer strains of American origin became positive occasionally at rather higher age than in the strains of Swiss origin. DD/Tbr, KF and DDD strains expressed MTV antigen in the male genital organs at intermediate age between the strains of Swiss origin and American origin. Complete disappearance of MTV antigen expression in these organs of DDf strain indicates that the expression of MTV in DD/Tbr is due to a milk-factor. PMID- 6274727 TI - Geographical pathology of gastric carcinoma: histological types among Chileans. AB - Nine hundred and twenty-three Chilean cases of gastric carcinoma who had received surgical resection of the stomach were histologically classified. In both males and females, diffuse carcinoma was a common type among younger patients, while adenocarcinoma was predominant in elderly patients. Statistical analysis demonstrated identical type distributions in all age groups in the Chilean series, as was seen in other ethnic groups surveyed previously. PMID- 6274728 TI - Membranous obstruction of the inferior vena cava and hepatocellular carcinoma in South Africa. AB - This study was undertaken to establish the frequency of membranous obstruction of the inferior vena cava in South Africa, to characterize the pathology of the lesion and to define its relationship to hepatocellular carcinoma. Over a 9-yr period 101 cases of membranous obstruction of the inferior vena cava into the right atrium was occluded in all cases in which it was examined, and two basic patterns of the abnormality in the hepatic portion of the inferior vena cava have been described. The histologic picture in the liver was a chronic congestive fibrosis. In 44 cases of congestive fibrosis, diagnosed from liver biopsy specimens from black patients over a 5-yr period, vena caval membranes were demonstrated on cavography in 38 (86.4%). Hepatocellular carcinoma developed in 48 of the 101 cases (47.5%), indicating the importance of this defect in the pathogenesis of hepatocellular carcinoma in South Africa. PMID- 6274729 TI - Hepatotoxicity of vitamin A and ethanol in the rat. AB - To study the possible hepatotoxicity of vitamin A supplementation and its potentiation by ethanol, rats were fed diets with either normal or fivefold increased vitamin A content, both with or without ethanol. Ethanol with a normal vitamin A diet produced the expected proliferation of the smooth endoplasmic reticulum and moderate mitochondrial lesions. Vitamin A supplementation by itself produced endoplasmic reticulum proliferation, slight enlargement of mitochondria, and moderate decrease in cytochrome oxidase activity and cytochrome aa3 content. The combination of high vitamin A and ethanol resulted in much more striking lesions, with giant mitochondria containing paracrystalline inclusions and depression of oxygen consumption in state-3 respiration with five different substrates, including palmitate and palmitoyl coA. The depression of fatty acid oxidation may have contributed to the lipid accumulation. The blood levels of vitamin A were unaffected whereas liver levels of vitamin A were increased by vitamin A supplementation and decreased by ethanol. As a net result the liver vitamin A content of the high-A-ethanol groups was not greater than that of the normal-A-control group, suggesting that a metabolite of vitamin A rather than vitamin A itself may have been responsible for the potentiation of vitamin A toxicity by ethanol. Mitochondrial toxicity reflected itself also in decreased content of various cytochromes and reduced activity of enzymes, including glutamate dehydrogenase. The activity of the latter was increased in the serum. Implications of these findings for the routine treatment of alcoholics with vitamin A and the monitoring for possible signs of toxicity are discussed. PMID- 6274730 TI - Studies of the antisecretory activity of morphine in rabbit ileum in vitro. AB - We studied the influence of morphine on intestinal secretion induced by three different secretogogues in an attempt to elucidate the mechanism for its antisecretory activity. In rabbit ileal mucosa in vitro morphine (10(-4) M) did not influence the electrical response to the subsequent administration of prostaglandin E2 or of acetylcholine but did inhibit the net secretion of chloride provoked by both agents and prevented the reduction of sodium adsorption induced by the prostaglandin. Morphine reduced the peak electrical response to cholera toxin and reduced toxin-induced net chloride secretion. The modes of action of morphine appeared to be by both enhancing adsorption, as it does in control, nonsecreting mucosa, and by inhibiting secretogogue activity. PMID- 6274731 TI - Immunocytochemical localization of intrinsic factor--cobalamin bound to the guinea pig ileum in vivo. AB - The absorption of dietary cobalamin requires that a complex of intrinsic factor cobalamin bind to specific receptors in the distal small intestine. The cellular location of these binding sites is unknown. To localize the intrinsic factor cobalamin binding site, homogeneously purified hog intrinsic factor (7 pmol) and cobalamin (7 pmol) were instilled into ileal loops (n = 10) in anesthetized guinea pigs. Tissue controls for binding specificity were intrinsic factor cobalamin in colon (n = 3) and jejunal (n = 3) loops, as well as intrinsic factor alone in ileal loops (n = 3). Intrinsic factor-[57Co]cobalamin was instilled in three ileal loops maintained for 90 min, and the radioactivity in the liver and kidneys was measured. Portions of each loop were washed in Krebs-Ringer buffer + Ca++, pH 7.4, or in either 1 mM Na2 ethylenediaminetetraacetic acid or in pH 4.5 acetate buffer. All tissue was processed for ultrastructural immunocytochemistry using a monospecific antisera to hog intrinsic factor as part of a well characterized indirect immunoperoxidase method. Essentially all immunoreactive intrinsic factor-cobalamin was associated with the microvillous pits on mature ileal absorptive cells. Intrinsic factor binding was not present in colon or jejunum, or in the ileal loops instilled with intrinsic factor alone. Ethylenediaminetetraacetic acid and acid pH decreased intrinsic factor-cobalamin binding but caused severe disruption of the microvillous architecture. Absorption of [57Co]cobalamin was demonstrated, but intrinsic factor was not identified within enterocytes. The localization of specific intrinsic factor-cobalamin binding sites to the microvillous pit suggests that the intrinsic factor cobalamin receptor may maintain a relatively fixed position within the surface membrane, and although alternative explanations are possible, our findings support the hypothesis that intrinsic factor is not internalized into the enterocyte during cobalamin absorption. PMID- 6274732 TI - Cellular mechanisms of bile formation. PMID- 6274733 TI - Membranous obstruction of the inferior vena cava: etiology and relation to hepatocellular carcinoma. PMID- 6274734 TI - Biliary artresia and non-A, non-B hepatitis? PMID- 6274735 TI - Comparison of gallbladder contractions induced by motilin and cholecystokinin in dogs. PMID- 6274736 TI - [Assessment of prognostic criteria in uterine carcinomas (author's transl)]. AB - Prognostic criteria of cervical and uterine carcinomas can be important for both physician and patient. Compared with patients not subjected to surgical treatment, a multitude of prognostic criteria must be investigated in operated women via the systemic examination of the surgical specimen, reaching beyond the weight attached to the clinically assumed staging. These criteria enable quantitative and qualitative characterization of the carcinomas allowing individualisation of therapy adapted to the tumour. PMID- 6274737 TI - [Restrictase BamHI, HindII, EcoRI and SmaI mapping of hybrid lambda-phi 80 phages with recombination in the structural gene region]. AB - Hybrids lambda H lambda T80 with recombination in the region of structural genes have lambda head and phi 80 tail genes. In this paper the molecular structure of 5 independently isolated hybrids was established using restriction endonucleases. It has been shown that all of them have a recombinant head or tail. A deletion of 4,8% lambda was demonstrated in the immunity region of phi 80vir phage. Co ordinates of restriction sites for BamHI, HindIII, EcoRI and SmaI restriction endonucleases on phi 80 DNA were calculated. PMID- 6274738 TI - [Lambda H-lambda T 80 hybrid study of the DNA structural gene region in lambda and phi 80 phages]. AB - Hybrids lambda H lambda T80 are formed due to recombination of the phage lambda att80 and phi 80 prophage partially deleted in the region of structural genes. Genetic structure of 22 independently isolated lambda H lambda T80 hybrids was determined by the restriction method and it was shown that recombination took place in the genes A, C, D and H. The frequencies of hybrid formation diminish from 1.10(-3) to 4.10(-5) for this gene order, which suggests that the polar divergence of nucleotide sequencies in the region of structural genes exists. It was found that formation of hybrids with recombination in the region of "weak" homology (gene H) was possible only when the region of "strong" homology was present in the deleted phi 80 prophage to initiate recombination. PMID- 6274739 TI - Dietary fiber: is it really helpful? PMID- 6274740 TI - [Prevention of reservoir pollution from pesticide and mineral fertilizer residues originating with surface runoff]. PMID- 6274741 TI - [Sanitary improvement in the working conditions in deep open-cut mines]. PMID- 6274742 TI - [Intracutaneous 239Pu distribution after its application in nitric acid solutions]. PMID- 6274743 TI - [Changes in the peripheral blood and hemolytic activity of the dust in animals receiving beryllium and pure quartz dust]. PMID- 6274744 TI - Some recent trends in studies of human lymphoid cells: B-cells, Epstein-Barr virus, and transformation. PMID- 6274745 TI - Epstein-Barr virus: its site of persistence and its role in the development of carcinomas. PMID- 6274746 TI - Episomal and nonepisomal herpesvirus DNA in lymphoid tumor cell lines. AB - Tumor cell lines derived from Herpesvirus saimiri (H. saimiri)- and Herpesvirus ateles (H. ateles)-induced lymphomas of New World primates and rabbits contain multiple copies of viral genomes. Partial denaturation mapping and blot hybridizations of episomal DNA from lymphoid tumor cell line No. 1670 showed that a 12.5md-fragment is missing which represents the EcoRI D- and H-fragments of virion L-DNA. However, the missing piece can be demonstrated in total cellular DNA by reassociation kinetics, possibly because it persists in integrated form. Both episomal and nonepisomal H-DNA are heavily methylated in a number of the lymphoid cell lines, and methylation may be reduced by conventional methylation inhibitors (S-adenosyl homocystein, SIBA) as well as by the tumor promoting phorbol ester TPA. PMID- 6274747 TI - Immunopathology of the X-linked lymphoproliferative syndrome. AB - The immunopathogenesis of 25 kindreds affecting 100 males with the X-linked lymphoproliferative syndrome (XLP) is being studied comprehensively by our registry and laboratory group. XLP is a combined variable immune deficiency with Epstein-Barr virus (EBV) induced phenotypes of: (1) fatal infectious mononucleosis (IM), (2) chronic IM progressive to malignant lymphoma, (3) acute IM progressive to acquired agammaglobulinemia or (4) malignant lymphoma. Cytogenetic studies of peripheral blood lymphocytes from 15 surviving males and 21 carrier females reveal random karyotype errors in several kindreds. Often polyclonal Ig or selective IgM increases and lymphocytosis with plasmacytoid forms typifies the IM phenotypes. Weakly reactive EBV-specific antibodies are found and anti-EB nuclear antigen is lacking. Antibodies to EBV are paradoxically elevated in female carriers. Initially all lymphoid tissues show immunoblastic proliferation with plasma cell differentiation and focal to extensive necrosis. Thymus gland and other lymphoid organs become depleted in T cell regions and Hassall's corpuscles may become destroyed. Multinucleated giant cells may be seen destroying the corpuscles or calcified corpuscles are found. The lymphoid infiltrates and lesions resemble graft-versus-host response in the fatal IM phenotype. Extensive necrosis in lymph nodes and deficient Ig secretion of B cells characterize acquired agammaglobulinemia phenotypes. The malignant lymphomas span the spectrum of B cell differentiation with most being immunoblastic sarcomas. One case probably was monoclonal thus far, others are being studied. EBV DNA hybridization of tissues from 7 patients with fatal IM revealed 1 to 20 EBV genome equivalents per cell. The patients lacked appropriate EBV antibody responses. Our studies of XLP support the hypothesis that immune deficiency the EBV permits chronic and fatal lymphoproliferative diseases in XLP following EBV infections. Owing to this knowledge, rational bases for prevention by genetic counseling and by providing high titer gammaglobulin and antiviral therapy is being attempted. PMID- 6274748 TI - Human leukemic lymphocytes--biochemical parameters of the altered differentiation status. PMID- 6274749 TI - The relative role of viral transformation and specific cytogenetic changes in the development of murine and human lymphomas. PMID- 6274750 TI - Transforming genes of retroviruses: definition, specificity, and relation to cellular DNA. PMID- 6274751 TI - A cellular protein phosphorylated by the avian sarcoma virus transforming gene product. PMID- 6274752 TI - The transformation-specific protein pp60src from an avian sarcoma virus. PMID- 6274753 TI - Proteolytic processing of avian and simian sarcoma and leukemia viral proteins. PMID- 6274754 TI - Characterization of the hematopoietic target cells of defective avian leukemia viruses by velocity sedimentation and density gradient centrifugation analyses. PMID- 6274755 TI - PRCII, a representative of a new class of avian sarcoma viruses. AB - The Poultry Research Center Virus II (PRC II) is a replication-defective avian sarcoma virus with envelope determinants of the A and B subgroups. In nonproducing cells transformed by PRCII the products of the replicative genes gag, pol, and env are not demonstrable, but a single polyprotein of Mr 105,000 (p105) can be detected. P105 contains peptides of the gag proteins p19 and p27 plus transformation-specific sequences. It does not contain peptides of gPr95env of Pr180gag-pol (with the possible exception of one pol peptide). The transformation-specific sequences of p105 are distinct form those of p100 of avian carcinoma virus MH2, of p110 coded for by avian myelocytoma virus MC29, and of p75 or p40 of avian erythroblastosis virus AEV. They also show no resemblance to p60src of Rous sarcoma virus. P105 is phosphorylated on a tyrosine residue and has an associated phosphokinase activity. P105 appears to be capable of autophosphorylation and of phosphorylating homologous immunoglobulin. PMID- 6274756 TI - Genetics of leukemogenesis by avian leukosis viruses. PMID- 6274757 TI - Avian lymphoid leukosis is correlated with the appearance of discrete new RNAs containing viral and cellular genetic information. PMID- 6274758 TI - The molecular basis of avian retrovirus-induced leukemogenesis. PMID- 6274759 TI - Genetic approaches toward elucidating the mechanisms of type-C virus-induced leukemia. PMID- 6274760 TI - Ontogeny of Abelson murine leukemia virus target cells. PMID- 6274761 TI - Enhancing effect of murine leukemia virus on fibroblast transformation by normal BALB/c mouse DNA fragments. PMID- 6274762 TI - Role of viruses in the etiology of naturally occurring feline leukemia. PMID- 6274763 TI - Relationship of the feline oncornavirus associated cell membrane antigen to a feline sarcoma virus encoded polyprotein. PMID- 6274764 TI - Feline leukemia virus nonproducer lymphosarcomas of cats as a model for the etiology of human leukemias. PMID- 6274765 TI - Genomic integration of bovine leukemia provirus and lack of viral RNA expression in the target cells of cattle with different responses to BLV infection. PMID- 6274766 TI - Regulation of human T-cell proliferation: T-cell growth factor and isolation of a new class of type-C retroviruses from human T-cells. AB - The discovery, characterization, and purification of human T-cell growth factor (TCGF) has led to the establishment of continuously growing T-lymphoblast cell lines from normal people and from patients with certain T-cell neoplasias. In contrast to normal T-cells, neoplastic mature T-cells respond directly to TCGF, requiring no prior lectin or antigen in vitro activation. The transformed T-cell lines have phenotypic characteristics consistent with the neoplastic cells of their disease of origin. A novel retrovirus, human T-cell lymphoma-leukemia virus (HTLV), has been isolated from the fresh and cultured cells of two of these patients. Subsequent characterization of this virus has shown that it is not significantly related to any known animal retrovirus, is not an endogenous (genetically transmitted) virus of man, and so far has been associated only with fresh or cultured T-cells from patients with T-cell neoplasia. These results suggest that HTLV infected some mature T-cells of some people and that it might be involved in some neoplasias involving these cells. PMID- 6274767 TI - Viral gene expression in cells transformed by simian sarcoma virus, an infectious primate type C retrovirus. PMID- 6274769 TI - [Management of surgically treated breast cancer with special reference to hormone receptors and hormone therapy]. PMID- 6274768 TI - ELISA for the detection of antigens cross-reacting with primate C-type viral proteins (p30,gp70) in human leukemic sera. PMID- 6274770 TI - Multiple glomus tumours in one digit. PMID- 6274771 TI - [Effect of suloctidil on cerebral venous outflow in the dog (author's transl)]. AB - The effects of intravenous administration of suloctidil [erythro-1-(4 isopropylthiophenyl)-2-n-octylamino-propanol], (MY 103) on cerebral venous outflow (VOF), blood gases, cerebral metabolic rate of oxygen (CMRO2) and other hemodynamic parameters were studied and effects compared to those of cinnarizine and papaverine, in gallamine immobilized dogs. MY103 and papaverine at doses 0.1 1.0 mg/kg, i.v. caused significant increases in VOF by 6.1-38.3% and 12.1-26.1% respectively. The changes in VOF persisted for 1.4 min and 0.5 min in mean following 1.0 mg/kg, i.v. of MY103 and papaverine, respectively. Cinnarizine also increased VOF by 10.0 and 18.7% at doses 0.3 and 1.0 mg/kg i.v. The duration of VOF changes by cinnarizine 1.0 mg/kg, i.v., 2.4 min (mean), was no longer than the duration seen with the same dose of MY103 or papaverine. Mean blood pressure (MBP) decreased dose dependently with these drugs and consequently the cerebral vascular resistance (CVR) decreased 8.6-40.2% with MY103, 13.6-25.0% with cinnarizine and 16.1-39.5% with papaverine administration, respectively. MY103 and papaverine at a dose of 1.0 mg/kg i.v. did not affect pH, Pco2 and Po2 in arterial blood, but these two drugs did show a tendency to lower Pco2 and elevate Po2 in cerebral venous blood. CMRO2 was not significantly affected by MY103 or papaverine. It is suggested that cerebral vasodilation may be the mechanism for VOF increase following MY103 administration. PMID- 6274772 TI - Production of superoxide anion by phagocytosing human and pig leukocytes. PMID- 6274773 TI - [Organization of systematic after-care of cancer patients in a general hospital]. AB - For better care and control of our cancer-patients we have built up an organization for systematic follow-up. Two years' experiences are reported. After some difficulties in the beginning we now are able to follow-up and record the further course of the disease in all cancer-patients. Systematic oncologic follow up contributes to greater living quality of the patients and leads the physician always to compare the treatment with that of other clinics and hospitals; it also enhances interdisciplinary exchange and -last not least-scientific evaluation of the therapeutic results. PMID- 6274774 TI - [Assessment of the results of aorto-coronary bypass surgery with the technetium 99m pyrophosphate scintigram (author's transl)]. AB - In 40 patients with angiographically-documented coronary artery disease, technetium 99m pyrophosphate (99mTc-PYP) myocardial scintigrams were obtained prior to and four to six weeks after aorto-coronary bypass surgery. In the majority of patients, preoperative and postoperative exercise testing with simultaneous pulmonary artery pressure recordings was performed. In 22 of 30 patients with preoperatively increased 99mTc-PYP myocardial activity, no tracer accumulation could be found postoperatively. In the latter patients, there was also a significant increase in exercise capacity and lessening of ischemic ST segment depression in the exercise ECG. In the remaining eight patients in whom increased tracer accumulation was found to persist postoperatively, there was no improvement in exercise capacity. In ten patients with no myocardial tracer accumulation preoperatively, unchanged myocardial scintigrams and a significant decrease of the ischemic ST-segment depression in the exercise ECG were seen postoperatively except in one patient with perioperative myocardial infarction. These results were also partially confirmed by repeated coronary angiography and ventriculography. Patent bypass grafts were associated with regional improvement in left ventricular function. The results indicate that postoperative absence of myocardial tracer accumulation appears due to amelioration of preexisting myocardial ischemia at rest. Thus, in the assessment of the results of aorto coronary bypass surgery, the 99mTc-PYP scintigram offers an important diagnostic potential. PMID- 6274775 TI - [The experimental study of epidermal growth factor -effects of epidermal growth factor on the cyclic nucleotide system and protein phosphorylation in pig epidermis- (author's transl)]. AB - Incubation of pig skin slices with epidermal growth factor (EGF) caused an increase in intracellular cyclic GMP concentration of pig epidermis. A significant increase was found after 1 hr of incubation and reached a peak by 6 hrs. EGF caused no change in the cyclic AMP level, nor did it affect epinephrine induced cyclic AMP responses. EGF also stimulated protein phosphorylation in pig epidermis. The stimulation was both time and dose dependent, and was not inhibited by the simultaneous addition of cycloheximide. The addition of cyclic GMP did not mimic the EGF treatment and EGF stimulated protein phosphorylation in 20 minutes. Phosphoproteins stimulated by EGF were identified by SDS-PAGE and autoradiographic analysis. The identified phosphoproteins were fibrous keratin proteins (MW. 65,000, 60,000, 56,000 and 51,000) and soluble protein (MW. 30,000). Calcium ion potentiated the phosphorylation of keratin proteins by EGF. Ionophore A23187 plus calcium ion also enhanced the phosphorylation of keratin proteins. PMID- 6274776 TI - Bronchial cast shadow due to cellular impaction of the bronchi. AB - The genesis of unusual radiographic shadows by bronchi filled with tumor cells is reported. These shadows, similar to those caused by mucus or pus, are rod-like and indicate the replacement of air by water dense materials. The unifying term, "bronchial cast shadow" is proposed as the radiographic diagnostic sign. PMID- 6274777 TI - Intermittent stimulation with LH-RH in postmenopausal hypergonadotropism. AB - Ten postmenopausal women received 8 consecutive (1 hour interval) i.v. bolus injections of either 25 or 500 micrograms of LH-RH. Gonadotropin release was monitored before and during LH-RH stimulation. After the first LH-RH bolus a dose dependent LH release was seen. All consecutive LH-RH boli elicited similar LH responses resulting in pulse-type LH release. Throughout the whole stimulation period the pituitary gland remained highly responsive to LH-RH, FSH release was uncharacteristic. It is concluded that during postmenopause LH-RH is hyper secreted. Thus the pituitary gland remains responsive to repeated injections of even supraphysiological LH-RH doses. PMID- 6274778 TI - Influence of metabolic control on platelet functions in diabetic mellitus. AB - Two groups of diabetic patients in poor metabolic control were followed prospectively for platelet function in the course of adequate insulin therapy. All 8 ketoacidotic and 38 nonacidotic, but markedly hyperglycemic, patients showed decreased platelet aggregation at the time of entering the hospital in comparison to subsequent controls. Platelet adenylate cyclase activity rose during the observation periods significantly. Changes of beta-thromboglobulin plasma levels were not uniform. These data give evidence for a metabolic factor which modifies platelet functions to a considerable degree. PMID- 6274779 TI - Erythrocyte membrane microviscosity in diabetes. AB - Decreased erythrocyte membrane fluidity was revealed by analyses with fluorescence depolarization using 1,6-diphenyl-1,3,5-hexatriene (DPH) and with electron paramagnetic resonance using spin-labelled stearates (EPR). The decreased membrane fluidity measured by DPH appeared to be related to the increase in membrane or plasam free cholesterol to phospholipid (C/PL) mol ratios. The decreased membrane fluidity observed by EPR appeared to be located, at least in part, in a deeper portion of membrane lipid core and the change may not be derived principally from the effect of cholesterol. Increases of membrane sphingomyelin (+ lysophosphatidylcholine) and decreases of phosphatidylethanolamine were found in erythrocytes of diabetic patients. The contributing factors to the decreased erythrocyte membrane fluidity in diabetic patients were suggested to be not only an increase in membrane C/PL mol ratios but also multifactorial changes involving membrane phospholipid fractions and other complex factors. It is considered that the determinant factors of membrane fluidity are so complex that the decreased membrane fluidity observed with diabetic erythrocytes cannot be closely correlated with any single constituent measured. The decreased membrane fluidity observed in this study appears to be related to poor metabolic control and is often associated with diabetic microangiopathy. PMID- 6274780 TI - Thyroxine binding globulin and thyroid function tests in patients with hepatocellular carcinoma. AB - To determine the prevalence of elevated serum concentrations of thyroxine binding globulin (TBG) in patients with hepatocellular carcinoma (HCC) and the influence of the associated cirrhosis, TBG was measured in 39 patients with HCC, 22 with and 17 without cirrhosis, in 20 patients with cryptogenic macronodular cirrhosis but without HCC, and in 40 matched controls. The mean serum TBG concentration in the patients was 34.5 +/- 17.7 microgram per ml, compared to 21.4 +/- 6.8 microgram pr ml in controls and 20.5 +/- 6.3 microgram per ml in cirrhosis without HCC (p less than 0.01). The presence or absence of cirrhosis in the HCC patients did not significantly influence the frequency with which elevated TBG levels were found; levels were normal in every subject with cirrhosis and no HCC. The mean thyroxine (T4): TBG ratio was 5.58 +/- 1.78 in controls and was reduced in HCC patients with both elevated (3.33 +/- 0.80, p less than 0.001) and normal TBG values (4.39 +/- 1.90, p less than 0.05), and in cirrhotics without HCC (4.29 +/- 1.01, p less than 0.01). T4 and TBG concentrations correlated significantly in controls, in HCC patients with elevated TBG, and in the cirrhotics without HCC. It is concluded that in patients with HCC (i) TBG levels may be elevated both in the presence or absence of cirrhosis; (ii) there is reduced binding of T4 TBG, and (iii) a low T4:TBG ratio excludes the diagnosis of hyperthyroidism in the presence of high T4 levels. TBG levels are normal in patients with cryptogenic macronodular cirrhosis without HCC. PMID- 6274782 TI - Histological classification of regional lymph nodes in relation to postoperative survival in primary lung cancer. PMID- 6274781 TI - Evidence that glucagon stabilizes rather than activates mitochondrial functions in rat liver. AB - The present study is concerned with the question as to whether the acute treatment of intact rats or hepatocytes with glucagon and dibutyryl cAMP, respectively, leads to a stabilization or an activation of mitochondrial functions, such as state-3 respiration, succinate dehydrogenase activity and pyruvate carboxylase activity. For this purpose, the influence of various parameters of mitochondria preparation (isolation medium, washing steps, storage) as well as of phospholipase A inhibitors (cinchocain, chloroquine) on the expression of the hormone effect was examined. With regard to the above mentioned functions, the values displayed by control mitochondria were found to be considerably higher if mannitol instead of sucrose had been used for isolation. Accordingly, only small effects of hormone treatment became apparent. The addition of cinchocain or chloroquine to the sucrose medium yielded results similar to those obtained with mannitol. Furthermore, the hormone effect on state 3 respiration and succinate dehydrogenase activity was only small if the mitochondria had been prepared faster than usual and had been used without washing. Regarding pyruvate carboxylase, a considerably smaller glucagon effect was observed when it was assayed at 25 degrees C and not (as usual) at 37 degrees C. Our results indicate that glucagon application stabilizes rather than activates mitochondrial functions. PMID- 6274783 TI - Primary adenocarcinoma of an anal gland with secondary perianal fistulas. AB - A case of primary mucinous (colloid) adenocarcinoma associated with multiple perianal fistulas and a supra-anal intergluteal abscess of a short duration is described. The origin of this tumor has been much debated. Some believe that the tumor occurs primarily in an anal duct or its glands with secondary fistula formation, whereas others claim that the tumor arises in a chronic fistulous tract. The present study substantiates the former concept on the basis of the mode of clinical presentation and the pathologic findings of a large tumor, without involvement of the anorectal mucosa. The residual anal ducts and glands were found in close relationship with the tumor. PMID- 6274785 TI - Modulation of the time relation of action potential impulses propagating along an axon. PMID- 6274784 TI - A postmortem study of malignant glucagonoma with heart muscle hypertrophy, including chemical, histochemical, immunohistological and ultrastructural observations. AB - The autopsy findings in a 58 year old female with the typical glucagonoma syndrome with heart muscle hypertrophy are described. Despite remission of a skin rash and hyperglucagonemia following removal of metastatic liver tumors, the patient died of massive cerebral hemorrhage caused by uncontrolled hypertension and septic cerebral angiitis. On the basis of surgical and autopsy materials, the tumors in the pancreas and liver were defined as malignant glucagonoma by histological, histochemical, immunohistochemical, and ultrastructural studies. The pathohistological changes in several organs that might be related to the hypersecretion of pancreatic glucagon are discussed. PMID- 6274786 TI - Alteration in sodium channel gate kinetics of the Hodgkin-Huxley equations applied to an electric field model for interaction between excitable cells. PMID- 6274787 TI - Reversible ultrastructural changes in human fibroblasts grown in hepes buffered MCDB-104 supplemented with human serum. AB - Prolonged maintenance of human dermal fibroblasts in MCDB-104 medium supplemented with pooled human serum or platelet factor deficient preparations of human serum led to appearance of a large number of membrane bound inclusions, resembling lysosomes, and proliferation of small, Golgi associated vesicles. These inclusions did not appear if the cells were grown in Dulbecco-Vogt's modification of Eagle's minimal essential medium or in minimal essential medium supplemented with the same human serum fractions. Cells that acquired inclusions during a 10 d incubation in MCDB-104 subsequently lost these inclusions when transferred to Dulbecco'Vogt's medium for 4 d. Similar reversal of effects of MCDB-104 was also produced by MCDB-104 buffered with bicarbonate instead of HEPES. PMID- 6274789 TI - Covert infection of a mouse mammary tumor cell line with Mycoplasma hyorhinis: cosedimentation with mouse mammary tumor virus in sucrose density gradients. AB - Supernatant fluids from cultures of a mouse mammary tumor (MT) cell line were found to produce a specific cell detachment effect when inoculated into HeLa cells. The cell detachment factor (CDG) responsible for this effect was examined. Repeated attempts to cultivate this CDF in bacteriologic, fungal, and mycoplasma media were unsuccessful. However, using the DNA fluorochrome staining technique and specific immunofluorescent staining procedures, the CDF was identified positively as a noncultivable strain of Mycoplasma hyorhinis. It was also noted that this CDF could be labeled with [3H]uridine in MT cell cultures, concentrated, and banded at a density of 1.18 g/cm3 when centrifuged to equilibrium in a 20 to 60% sucrose gradient. Using a multiple antibiotic treatment regimen, the MT cells were "cured" of the M. hyorhinis contaminant. Re infection of these cells with an exogenous strain of M. hyorhinis resulted in the same cell detachment effect, and this strain when labeled with [3H]uridine also sedimented at a density of 1.18 g/cm3. The salient feature of these studies is that M. hyorhinis sediments at the same density of mouse mammary tumor virus (MMTV) in sucrose density gradients. This was demonstrated by sucrose density gradient analyses of a purified sample of MMTV, assaying for reverse transcriptase activity, and a [3H]uridine labeled sample of the M. hyorhinis present in the MT cell cultures. PMID- 6274788 TI - Development of two cloned epithelial cell lines from normal adult mouse and rat ventral prostates. AB - Two epithelial cell lines were established, one from adult C3H mouse and one from adult Fischer rat ventral prostate. These cell lines were obtained from explant cultures, using Ham's F12 medium supplemented with HEPES, insulin, testosterone, hydrocortisone, epidermal growth factor, and 7.5% fetal bovine serum. A low concentration of trypsin and EDTA in Ca++- and Mg++-free phosphate buffer was used for passaging the cells. The rat cell line was established following implantation of prostate tissue in nude mice. These cell lines stained positively for acid phosphatase and were dependent upon epidermal growth factor for growth. Morphological studies, including electron microscopy, revealed a highly characteristic epithelial morphology of both cell lines. These cell lines have hypotetraploid chromosome number and are capable of metabolizing benzo(a)pyrene. We propose the application of these cells as models for the study of prostate carcinogenesis. PMID- 6274790 TI - Carbohydrate surface constituents of T cells mediating delayed-type hypersensitivity that control entry into sites of antigen deposition. AB - Peritoneal exudate T lymphocytes (PETLs) that mediate delayed-type hypersensitivity (DTH) to sheep red blood cells in mice were modified by in vitro treatment methods which modify surface carbohydrate constituents. Neuraminidase treatment resulted in the release of both N-acetylneuraminic acid and N-glycolyl neuraminic acid, and periodate treatment in the formation of the corresponding C7 analogues. Treatment of PETLs with neuraminidase led to a transient reduction of DTH reactions in syngeneic cell recipients. After treatment with neuraminidase plus galactose oxidase, adoptive mediation of DTH was more markedly reduced. Incubation of PETLs with periodate caused a permanent loss of DTH transferring capacity. The oxidation-induced effects following treatment of PETLs with neuraminidase plus galactose oxidase or with periodate could be reversed by subsequent reduction with borohydride of the previously formed aldehyde moieties. Decreased DTH reactions observed after the various treatment procedures were paralleled by reduced immigration of PETLs into sites of antigen deposition, indicating that the reduction/oxidation state of cell surface carbohydrates is crucial for induction of inflammatory processes by T cells. Trapping of PETLs in the liver could not be the sole mechanism since neuraminidase- and periodate treated PETLs, but not neuraminidase plus galactose oxidase-treated PETLs, accumulated in the liver. It therefore appears that alterations in the reduction/oxidation state of the cell surface can lead to unresponsiveness of T cells to inflammatory signals. PMID- 6274791 TI - Enhanced immune reactivity to hepatoma 22a cells in MTV-infected compared with MTV-free C3H mice. AB - The transplantability and growth rate of mouse mammary and non-mammary tumours (sarcoma and hepatoma) were compared in MTV-free (MTV-) and MTV-infected (MTV+) C3H mice. It was demonstrated that sarcoma and hepatoma, in contrast to mammary tumours, grew better in MTV- mice. The difference between C3H sublines in the growth of hepatoma transplants was found to be determined by the stronger specific immune response and higher natural lymph node cell cytotoxicity in MTV+ mice. PMID- 6274792 TI - Effects of trifluoperazine on human neutrophil function. AB - The interaction of cytochalasin B-treated human neutrophils with the synthetic tripeptide, N-formyl-methionyl-leucyl-phenylalanine (FMLP) results in a time- and concentration-dependent generation of superoxide anion (O2-) by an extracellular release of granule-associated beta-glucuronidase and lysozyme from these cells. Granule exocytosis was not accompanied by significant cytoplasmic lactate dehydrogenase extrusion. FMLP-stimulated O2- production occurs but is significantly curtailed in the absence of extracellular calcium. Nevertheless, incubation of neutrophils with EGTA in calcium-free medium had no effect on the O2- -generating system. Trifluoperazine (TFP), an inhibitor of calmodulin (a calcium-binding protein), caused a dose-related inhibition of FMLP-elicited degranulation and O2- production in the presence of absence of extracellular calcium. This effect TFP could be reversed by washing the cells before contact with FMLP. These data suggest that TFP represents a useful tool for defining the relevance of calmodulin and calcium to neutrophil function. PMID- 6274793 TI - The role of accessory cells and T cell-growth factor in induction of cytotoxic T lymphocytes against herpes simplex virus antigens. AB - The roles of accessory cells and T cell-growth factor (TCGF) in the in vivo induction of herpes simplex type 1 (HSV) specific cytotoxic lymphocytes (CTL) were evaluated. Spleen cells from animals infected with HSV 4-6 weeks previously were depleted of adherent cells by passage over Sephadex G10. Unlike intact cells, such depleted spleen cells failed to respond by producing H-2 restricted virus-specific CTL upon culture for 5 days with infectious HSV. The CTL response could be restored either by adding normal genetically compatible peritoneal cells as accessory cells or by the addition of TCGF. To obtain optimum restoration accessory cells needed to be added soon after culture initiation but with TCGF addition, partial restoration was evident when added as late as 72 hr after culture. TCGF also permitted intact spleen cells to respond to heat-inactivated virus. The results are interpreted to indicate that accessory cells are essentially required for the presentation of virus to specific helper cells with such cells responding by the production of TCGF. The results also indicate that certain forms of virus may trigger the response of CTL precursors but not the response of helper cells. PMID- 6274794 TI - Augmentation of the antibody forming cell response to neuraminidase-treated cells by myxoviruses. AB - Following the inoculation of Newcastle Disease virus, Sendai virus and influenza virus CBA mice made a specific plaque-forming cell response to virus-treated sheep red blood cells but not to virus-treated L1210 cells. This non-specific response to virus-treated L1210 could be exactly reproduced by treatment of L1210 with bacterial neuraminidase. Neuraminidase-treated cells from eight of nine vertebrates including chickens and syngeneic mice also registered plaques. Inoculations of the above viruses were considered to have evoked separate antibody forming cell responses to viral antigens, foreign cell components in the virus inoculum and to neuraminidase revealed cell membrance antigens. PMID- 6274796 TI - Nephroblastomas-morphologic assessment based on 36 cases. PMID- 6274795 TI - Polymorphonuclear leucocyte membrane-stimulated oxidative metabolic activity-- the effect of divalent cations and cytochalasins. AB - The effect of divalent cations and cytochalasins on the oxidative metabolic response of polymorphonuclear leucocytes (PMNL) following membrane stimulation by opsonized zymosan, phorbol myristate acetate (PMA), digitonin and n-formyl methionyl-leucyl-phenylalanine (FMLP) was investigated using several techniques. For optimal ferricytochrome C reduction, oxygen consumption, luminol- and lucigenin-dependent chemiluminescence by PMNL on exposure to opsonized zymosan, extracellular magnesium was necessary. In contrast FMLP- and digitonin-induced PMNL metabolic activities were less dependent on extracellular magnesium than on calcium. Phorbol myristate acetate-induced PMNL oxidative metabolic activity occurred independently of extracellular magnesium and calcium concentration. Incubation of PMNL with cytochalasins B and E had little effect on PMA-induced metabolic activity, enhanced FMLP-induced metabolic activity but inhibited digitonin-induced metabolic activity. Investigation of zymosan-induced PMNL metabolic activity after incubation with cytochalasins demonstrated a dissociation between the different parameters of metabolic activity measured. The results described in this study support the suggestion that PMNL oxidative metabolism can be activated by at least two different mechanisms depending on the stimulus used, and that these mechanisms can in part be differentiated by their dependence on extracellular divalent cations. PMID- 6274798 TI - Rickettsia sennetsu in human blood monocyte cultures: similarities to the growth cycle of Ehrlichia canis. AB - Microscopic examination of cultured human monocytes infected with Rickettsia sennetsu and stained by the Giemsa method revealed the presence of various organismal growth forms in the cytoplasm of the infected cells. The growth forms were loosely scattered individual organisms, clusters of organisms, various sizes of dense inclusion bodies in intact and vacuolated cytoplasm, and organisms in close proximity to disintegrated monocytes. The appearance and the morphology of these R. sennetsu growth forms were similar to those of Ehrlichia canis propagated in canine monocytes. Specific identification of R. sennetsu was made by staining cultured monocytes with fluorescein-conjugated globulins extracted from the pooled sera of patients recovering from sennetsu rickettsiosis. Mice infected with cultured R. sennetsu developed gross pathological changes indicative of infection, and the organism was demonstrated in their spleens, peritoneal macrophages, and mononuclear blood cells. Human monocyte culture appeared to be more sensitive than the previously used African green monkey kidney cell line (BSC-1) for the isolation of R. sennetsu from samples containing minute infectious quantities of this organism. PMID- 6274797 TI - Permissiveness of rabbit monocytes and macrophages for herpes simplex virus type 1. AB - The permissiveness of rabbit monocytes and macrophages for herpes simplex virus was examined. Peripheral blood monocytes, alveolar macrophages, and peritoneal exudate macrophages were studied for their ability to replicate herpes simplex virus strains RE and KOS. Results indicated different degrees of interaction with virus depending on the macrophage type. Only peritoneal exudate macrophages showed evidence of virus replication. Productive infection was limited, with only a small number of cells (0.02%) yielding infectious virus. Higher numbers of cells appeared to be abortively infected. Approximately 40% expressed antigens, whereas virtually all were killed by exposure to virus. Coreless particles were seen by electron microscopy in about one-third. Alveolar macrophages were also killed by virus and showed evidence of virus adsorption, but showed no indication of productive or abortive infection. Monocytes neither adsorbed nor replicated virus, and viability was unaffected. Results suggest that differences in degrees of cellular maturation or differentiation, or both, account for the spectrum of interactions seen between herpes simplex virus and rabbit macrophages. PMID- 6274799 TI - Influence of sublethal antibiotic concentrations on bacterial adherence to saliva treated hydroxyapatite. AB - The influence of growth in the presence of sublethal concentrations of nine antibiotics on the ability of certain potentially odontopathic bacteria to attach to saliva-treated hydroxyapatite surfaces which mimic teeth was studied. Cells of Actinomyces viscosus LY7 and S2, Bacteroides gingivalis 381, Capnocytophaga ochraceus 6, and Actinobacillus actinomycetemcomitans N27 attached in lower numbers to saliva-treated hydroxyapatite when grown in the presence of 50% of the minimum inhibitory concentration of tetracycline. Electron microscopic observations of negatively stained preparations indicated that tetracycline-grown A. viscosus LY7 cells had fewer fimbriae than did untreated cells, which may account for the impaired ability of the treated cells to attach. However, cells of Actinomyces naeslundii L13 and S4 attached in higher numbers when grown in the presence of tetracycline, clindamycin, erythromycin, chloramphenicol, or neomycin. Streptococcus mutans strains H12 and JBP also exhibited increased adherence to saliva-treated hydroxyapatite when grown in the presence of 50 or 25% of the minimum inhibitory concentration of penicillin. Thus, growth in the presence of sublethal antibiotic concentrations could increase as well as decrease the adherence of bacteria to saliva-treated hydroxyapatite. Antibiotic grown cells of the Actinomyces strains showed enhanced hemagglutination activity, but this did not correlate with their ability to attach to saliva-treated hydroxyapatite. Sublethal concentrations of antibiotics in the growth media also affected the coaggregation reactions of several organisms; the effects were specific for one member of the coaggregation pair. PMID- 6274800 TI - In vitro infection of murine macrophages with Junin virus. AB - Mouse peritoneal macrophages were successfully infected with two strains of Junin virus producing high titers with no apparent cell damage. Infected cultures survived longer than noninfected cultures. The pattern of virus release suggested a persistent infection. Virus replication was delayed in macrophages from mice previously immunized with Junin virus. These results support the opinion that macrophages are targets for virus replication in vivo infections. PMID- 6274801 TI - Persistent infection with bovine herpesvirus type 1: rabbit model. AB - Persistent infection with bovine herpesvirus type 1 (BHV-1) was established in all rabbits after conjunctival inoculation of virus. Spontaneous reactivations of BHV-1 with and without the appearance of recurrent ocular lesions were observed in persistently infected rabbits. BHV-1 was reactivated predictably and shed from all persistently infected rabbits after the administration of dexamethasone. During all reactivations, BHV-1 isolation was restricted to the inoculated eye. PMID- 6274802 TI - Mode of action of staphylococcal leukocidin: effects of the S and F components on the activities of membrane-associated enzymes of rabbit polymorphonuclear leukocytes. AB - The cytotoxic action of the S component of leukocidin from Staphylococcus aureus on rabbit polymorphonuclear leukocytes was supported by the following observations, (i) Leukocytes displayed a large chemotactic response to the S component (10(-10) M) as well as to the chemotactic factor N formylmethionylleucylphenylalanine (10(-11) M). (ii) The S component stimulated high levels of phospholipase A2 activity in the cell membranes, with concomitant synthesis and release of prostaglandins. (iii) Uptake of 45Ca into leukocytes exposed to the S component was about double the rate of uptake into untreated cells. The increased 45Ca uptake into the cells was not inhibited by trifluoperazine and ruthenium red. (iv) Indomethacin and alloxazine, which had no effects on the binding of the S component to the cells, attenuated markedly the stimulation of phospholipase A2 activity, the syntheses of prostaglandins, and the increased uptake of 45Ca caused by the S component. The F component of leukocidin, bound to rabbit leukocytes with the aid of the S component, rapidly induced complete release of 86Rb from preloaded leukocytes. This release resulted from stimulation of ouabain-insensitive (Na+ + K+)-adenosine triphosphatase activity and inhibition of cyclic AMP-dependent protein kinase. PMID- 6274803 TI - Chemostat studies of the effect of environmental control on Streptococcus sanguis adherence to hydroxyapatite. AB - Streptococcus sanguis is a major component of early dental plaque. The ability of S. sanguis to adhere to salivary pellicle appears to involve specific bacterial surface receptors. The nature of these receptors is still not known; however, the component molecules may be subject to environmental control as has been shown for teichoic acids and certain proteins. To study these environmental effects, a chemostat was employed to vary the growth conditions of Streptococcus sanguis strain G9B. This strain has been used extensively to study the adhesion of [(3)H]thymidine-labeled batch-grown cells to saliva-coated hydroxyapatite beads. The effects of dilution rate, pH, and carbon source on adhesion were studied with a competition assay in which the labeled batch cells were used as a reference standard. In this assay, cells from the chemostat were harvested and compared for their ability to inhibit adhesion of labeled cells relative to unlabeled control batch-grown cells. Subsequent studies used chemostat grown cells labeled with [(3)H]thymidine as a reference standard so that results were internally controlled and reflected only the particular alteration in environment which was studied. These results indicated that when glucose was used as a growth-limiting substrate, cells grown at relatively high dilution rates (D = 0.5 h(-1); mean generation time = 1.4 h) behaved similarly to batch-grown cells and appeared to compete for the same binding sites. Cells grown at D = 0.1 h(-1) (mean generation time = 7 h) no longer competed with either batch-grown cells or chemostat cells grown at D = 0.5 h(-1). Moreover, adsorption isotherms of such slow-growing cells (D = 0.1 h(-1)) suggested that binding was no longer specific. When fructose was used as the growth-limiting carbohydrate, cells grown at D = 0.1 h(-1) did not show this loss of specificity and competed nearly as well as control batch-grown glucose cells. However, the effect of pH appeared to be independent of carbohydrate source, because cells grown in either glucose or fructose at pH 5.5 at D = 0.1 h(-1) lost the ability to compete with reference batch or chemostat cells grown at D = 0.5 h(-1). This effect was very sharp, since cells grown in the pH range from 6 to 7.5 at D = 0.5 h(-1) competed nearly as well as control cells. A similar effect of pH was found for batch cultures grown with excess glucose. These studies reinforce the idea that the environment can profoundly affect the bacterial surface and consequently the ability of the organism to adhere, a property which appears to be a primary event in some infectious diseases and in dental plaque formation. PMID- 6274805 TI - Fine structure and FSH binding of Sertoli cells in the blue fox (Alopex lagopus) in different stages of reproductive activity. AB - A clearly different Sertoli cell morphology was found in the immature blue fox and in the adults in and out of season. Immature cells had small, basally, located nuclei rich in peripheral heterochromatin, and few cytoplasmic organelles. Sertoli cells from adults in season had basally located, large, convoluted nuclei with homogenous nucleoplasma and prominent nucleoli. The basal and intermediate parts of the cytoplasm contained an extensively developed ER, numerous mitochondria, free ribosomes, lipid droplets and residual bodies, while the apical cytoplasm showed few distinct structures. In Sertoli cells from adults out of season the nuclei were dislocated towards the lumen, and apart from numerous dilated cisternae of ER, there were generally fewer organelles, but more glycogen particles. A marked rise in FSH binding was found towards the breeding season. PMID- 6274804 TI - Characteristic differences between saliva-dependent aggregation and adhesion of streptococci. AB - Comparison of saliva-mediated aggregation of Streptococcus sanguis, Streptococcus mitis, and Streptococcus mutans and adhesion of these organisms to saliva-coated hydroxyapatite showed that there was no relationship between these two activities. Adsorption of salivary aggregating activity to bacteria appears to have little effect on the ability of the residual saliva to support adherence; conversely, adsorption of salivary adherence factors to hydroxyapatite does not affect aggregation. Although heating saliva significantly reduces bacterial aggregation, it has little or no effect on adherence. A comparison of aggregation and adhesion with serial dilutions of saliva demonstrated that adhesion could still be detected at 100 to 500-fold-lower concentrations of salivary protein that bacterial aggregation. These findings support the concept that aggregation and adherence involve two distinct mechanisms of microbial clearance in the oral cavity. PMID- 6274806 TI - Biological studies with low and high molecular weight inhibin preparations. AB - The effects of both high and low molecular weight inhibin preparations on testicular and pituitary receptors were studied. Both these preparations were able to inhibit the binding of labelled hFSH to testicular receptor in a dose related manner, but were unable to affect the binding of labelled hCG to its receptor, suggesting that the observed inhibition of FSH binding was specific to inhibin. In addition, the binding of LHRH to pituitary receptors was affected by inhibin preparations. Interestingly, the antiserum raised against high molecular weight inhibin was able to neutralize, totally, the biological effect of high molecular weight inhibin, and only, partially, the biological effect of low molecular weight inhibin. PMID- 6274807 TI - Nerve conduction times in uremia. PMID- 6274808 TI - Influence of the motor unit action potential morphology (MUAP) on the EMG signal: theoretical and experimental study. AB - The aim of this work is the study of the influence of the MUAP morphology on the compound EMG signal by means of a simulation model. The simulated EMG is obtained as the sum of a number of MUS chosen on the basis of considerations concerning the muscle fiber dimension, the concentric needle electrode uptake area and the conduction volume. Myopathic and neurogenic conditions have been simulated together with the normal case, adopting typical MUAPs. The interspike interval (ISI) has been simulated by means of a stochastic variable. Both time and frequence domain analysis have been performed in the simulated EMG. The parameters obtained have been compared with those obtained from the experimental EMG in corresponding groups of pathologies. The model has provided clarifying data concerning the influence of the morphology of the MUAP, confirmed by the experimental findings. Simulation of abnormalities in the firing pattern as well as in the recruitment, seems to be necessary in order to improve the prediction capabilities of the model. PMID- 6274809 TI - Induction of Epstein-Barr virus by an new tumor promoter, teleocidin, compared to induction by TPA. AB - The effect of teleocidin, a new, naturally occurring tumor promoter on induction of Epstein-Barr virus (EBV), was compared with that of a known tumor promoter, 12 O-tetradecanoyl-phorbol-13-acetate (TPA). Early antigen (EA) and/or capsid antigen (VCA) of EBV was induced in the EBV genome-carrying cell lines C-6 and P3HR-I cells by teleocidin, its effect being maximal at a concentration of 12.5 ng/ml. The production of infectious EBV from P3-HR-I cells was enhanced by teleocidin maximally at a concentration of 0.5 to 2.5 ng/ml. The outgrowth of EBV transformed cells from peripheral lymphocytes of seropositive healthy donors was also enhanced by teleocidin at a concentration of 0.02 to 0.5 ng/ml. TPA tested simultaneously in all experiments exhibited the same activities as teleocidin, and was effective at similar concentrations. Teleocidin enhanced both EA and VCA synthesis in P3HR-I cells additively with n-butyrate, but not with TPA. This suggests that teleocidin and TPA have a common mechanism of action, although their chemical structures are different. PMID- 6274810 TI - Establishment of human cytotoxic T-cell lines specific for Epstein-Barr virus transformed autologous cells. AB - EBV-specific cytotoxic T cells (Tc) induced in vitro have continuously proliferated in vitro for over 9 months. The long-term maintenance of the Tc growth was dependent on periodic supplementation of both irrediated EBV transformed autologous lymphoblastoid cell line (LCL) cels and conditioned medium. The latter was derived from supernatants of human spleen-cell cultures stimulated with phytohemagglutinin. The cultured Tc maintained significant cytotoxic activity to autologous LCL cells but not to EBV-unrelated target cells, including K-562, B-, T-, null-cell lines and mitogen-stimulated antologous peripheral blood lymphocytes. Thus, established EBV-specific Tc lines from five different individuals always exhibited highly significant cytotoxicity against exhibited highly significant cytotoxicity against autologous LCL cells but not always against allogeneic LCL cells. Furthermore, restriction of the Tc to the autologous LCL was more pronounced after long-term culture than it was initially. This suggests that certain clones of Tc which arignificant cytotoxicity against exhibited highly significant cytotoxicity against autologous LCL cells but not always against allogeneic LCL cells. Furthermore, restriction of the Tc to the autologous LCL was more pronounced after long-term culture than it was initially. This suggests that certain clones of Tc which arignificant cytotoxicity against exhibited highly significant cytotoxicity against autologous LCL cells but not always against allogeneic LCL cells. Furthermore, restriction of the Tc to the autologous LCL was more pronounced after long-term culture than it was initially. This suggests that certain clones of Tc which are probably restricted to HLA are selectively established during long-term cultivation. PMID- 6274812 TI - Studies on lung tumours. V. Susceptibility of mice to dimethylnitrosamine-induced tumour formation in relation to H-2 haplotype. AB - Male mice of the strain A/Sn, of its congeneic partner strain A.SW, C57BL/10ScSnA (abbreviated: B10), and of two congeneic strains on a B10 background (B10.A and B10.AKM) were investigated for their susceptibility to lung tumour induction by dimethylnitrosamine (DMN) administered either by intraperitoneal injection or in the drinking water. Strain B10 (haplotype H-2b) proved to be very resistant, whereas strains A/Sn (H-2a) and A.SW (H-2s) were highly susceptible. The introduction of the haplotypes H-2a and H-2m in the resistant strain B10 resulted in a significant increase in sensitivity towards DMN-induced lung tumour formation. Lung tumour incidences in male (B10.A x B10)F hybrids, receiving DMN in drinking water, were found to be intermediate between, and significantly different from, the incidences of identically-treated parent strains B10.A and B10. Males of back-cross (B10.A x B10) x B10 BC proved to be of low susceptibility to lung tumour formation by DMN, tumour incidence being very low and not significantly different from that observed in identically-treated B10 males. It is concluded that, at least in the model system of B10-derived congeneic strains, H-2 haplotype is one of the factors important in determining susceptibility towards DMN-induced lung tumours. Comparison of C3Hf (H-2k), C3H/Sn (H-2k) and the latter's congeneic strains C3H.B10 (H-2b) and C3H.NB (H-2p) of male mice showed that these strains were moderately susceptible to both lung tumour and hepatoma formation by DMN. Accordingly, the presence of an H-2 haplotype from a lung-tumour-resistant strain (H-2b, B10) on the background of a strain of intermediate susceptibility (C3H) does not decrease susceptibility to lung tumour formation. The results were considered in the light of the H-2 haplotype dependence of spontaneous lung tumours, and consequently attention has been paid to the histological types of the induced and spontaneous lung tumours. PMID- 6274811 TI - Comparison of various serological and direct methods for the diagnosis of BLV infection in cattle. AB - The sensitivities of several serological and direct methods for the detection of bovine leukemia virus (BLV) infection in cattle have been critically compared. Among the serological methods, the radioimmunoassay (RIA) using the BLV glycoprotein (gp) was found to be more sensitive than the RIA using the internal virion antigen (p25) and the immunodiffusion test using the virion gp antigen. The differences in sensitivity between these tests were particularly evident with sera of cattle in early stages of infection. The sensitivities of the syncytia induction assay and the competitive RIA for the direct demonstration of BLV infection in cattle are comparable. Following natural infection, antibodies to the gp antigen in most cattle were detected by the RIA several months before infectious BLV became detectable by the SIA in the blood lymphocytes. It is concluded that the most accurate, sensitive, and rapid method for the detection of BLV infection in cattle is the RIA using the virion gp antigen. The data presented confirm that BLV infection as well as the antibody responses of naturally infected cattle to the BLV p25 and gp antigens are persistent. PMID- 6274813 TI - Natural killer cell activity in a population of leukemia-prone wild mice (Mus musculus). AB - Natural cell-mediated cytotoxicity against YAC-I targets was measured in splenocytes from leukemia-prone wild mice trapped near Lake Casitas (LC) in southern California. Cytotoxicity was mediated by cells that were non-adherent to nylon wool, non-phagocytic and resistant to thy-1.2 antiserum plus complement. Natural MuLV viremia in LC mice did not impair splenic cytotoxicity against TAC-I target cells, Cells infected with amphotropic and ecotropic MuLV of wild mouse origin were not appreciably lysed by LC splenic effectors. Although variable levels of cytotoxicity were detected against TAC-1 by normal spleen cells, consistently low levels of cytotoxicity against allogenic LC lymphoma, sarcoma and carcinoma targets were found using the same splenocytes. These results indicate that LC mice possess splenocytes with the characteristics of natural killer (NK) cells as defined in inbred mice. The resistance of LC-derived targets to lysis by LC NK cells suggests that NK cells may not be involved in natural tumor immunosurveillance or that the development of spontaneous tumors may involve escape from NK-mediated effector mechanisms. PMID- 6274815 TI - The formation and persistence of benzo(a)pyrene metabolite-deoxyribonucleoside adducts in rat skin in vivo. PMID- 6274814 TI - Association of Epstein-Barr virus with nasopharyngeal carcinoma in Alaskan native patients: serum antibodies and tissue EBNA and DNA. AB - Biopsy specimens from Alaskan Native patients with nasopharyngeal carcinoma (NPC) and from other patients seen on the otolaryngology service were tested for Epstein-Barr virus-specific DNA and nuclear antigen (EBNA). Serum samples from both groups were tested for various EBV-related antibodies. EBV DNA and EBNA results were in agreement in 29 of 31 tissue specimens tested by the two methods. Ten of 11 biopsies containing NPC cells were positive for EBV DNA. Two NPC patients had biopsies that showed only atypical epithelium but were also positive for EBV DNA or EBNA. The other tissue specimens were negative except for biopsies from two patients: one with a parotid gland lymphoepithelial lesion; another with undifferentiated carcinoma of salivary gland origin. PMID- 6274816 TI - Degeneration and regeneration of unmyelinated fibers in experimental leprous neuropathy. PMID- 6274817 TI - Cyclic nucleotides and lipolysis. AB - The recent literature regarding the mechanisms of regulation of lipolysis with emphasis on the role of cyclic nucleotides is reviewed. The following conclusions appear warranted at present. (1) Cyclic AMP (cAMP) is a major regulator of lipolysis. However, mechanisms other than the production and catabolism of cAMP also exist. (2) Insulin can lower adipocyte cyclic AMP levels, but this effect cannot explain all aspects of the antilipolytic effect of insulin. (3) Insulin stimulates cyclic AMP phosphodiesterase and inhibits adenylate cyclase in adipocytes. In addition, there are probably other targets of insulin action. The possibilities include cAMP dependent protein kinase, phosphoprotein phosphatase, and triacylglycerol lipase. (4) Cyclic GMP is probably not directly involved in the regulation of lipolysis. (5) Cytosolic Ca2+ probably plays an important role in the regulation of lipolysis. The nature of such a role for Ca2+ and the potential role of calmodulin in the regulation of lipolysis remain to be explored. PMID- 6274818 TI - Regulation of adipose-tissue lipolysis by phosphorylation of hormone-sensitive lipase. AB - Hormone-sensitive lipase has been purified from rat adipose tissue to almost 50 per cent protein purity and partially characterized. The isolated enzyme can be phosphorylated by ATP-Mg2+ in the presence of the catalytic subunit of cyclic AMP dependent protein kinase from the same tissue. Its activity towards emulsified triglyceride is thereby increased two-fold. The enzyme is phosphorylated also in the intact adipocyte, verifying the physiological relevance of the findings with the isolated enzyme. Noradrenaline causes a rapid increase in phosphorylation of the enzyme in intact adipocytes, immediately followed by a marked increase of its activity. Addition of dibutyryl-cyclic AMP to the adipocytes causes the same effects. The extent of phosphorylation of the enzyme after maximal noradrenaline stimulation of the adipocytes is rapidly decreased by insulin addition in close association with inhibition of the lipase activity. The results demonstrate that these hormones regulate the activity of the hormone-sensitive lipase, ie the rate of lipolysis in the adipocytes, by changes of the degree of phosphorylation of the enzyme. PMID- 6274819 TI - Adenosine and lipolysis. AB - The present review briefly summarizes work from the author's laboratory aiming at the clarification of the role played by adenosine in the regulation of adipose tissue circulation and metabolism. Studies on isolated fat cells demonstrate that adenosine, in mumolar concentrations, can inhibit lipolysis induced by eg noradrenaline. The effect appears to be due to inhibition of cyclic AMP formation. Studies of subcutaneous adipose tissue in situ also demonstrate the effect of exogenous adenosine on lipolysis. Furthermore, adenosine causes vasodilation and inhibition of noradrenaline release from sympathetic nerve endings. The actions of adenosine are antagonized by theophylline and other methylxanthines in concentrations lower than those required to inhibit cyclic AMP breakdown. It is shown that adenosine is normally present in adipose tissue in approximately 0.3 microM concentration and that the level is increased by sympathetic nerve stimulation. From a quantitative comparison of adenosine levels and dose-effect relationships as well as from pharmacological studies it is concluded that adenosine is a physiological regulator of adipose tissue circulation and lipolysis, while the physiological role of adenosine as a transsynaptic modulator of noradrenaline release remains to be established. PMID- 6274821 TI - Petechnetate-cimetidine abdominal imaging. PMID- 6274820 TI - Computerized scintigraphy of oesophageal bolus transit in asthmatics. PMID- 6274822 TI - Pyrophosphate scintigraphy: improved detection of the acute myocardial infarction with computer data manipulation and Fourier method analysis. PMID- 6274823 TI - Nosocomial viral infections: I. Epidemiology and significance. AB - Viral illnesses in Strong Memorial Hospital were monitored over a 17-month period. Using criteria based primarily on the incubation periods for a number of common virus infections, the infections we found were classified as hospital- or community-acquired. Hospital-acquired viral infections occurred on most hospital services; the majority of infections occurred on the pediatric and psychiatric services. Infections due to herpesviruses were seen more frequently in a group of patients aged 14 years or older, while infections in patients aged three years or younger were more likely to be due to respiratory syncytial virus, influenzavirus, adenovirus, or parainfluenza virus. Patients with nosocomial infections due to viruses were hospitalized an average of 9.3 days longer than uninfected controls; thus nosocomial viral infections result in increased costs of hospitalization. PMID- 6274824 TI - Scintigraphy in the diagnosis of beta cell carcinoma of the pancreas: case report. PMID- 6274825 TI - West Nile fever encephalitis. AB - Three cases of meningoencephalitis caused by the West Nile virus in young people are described. All patients had high fever, severe headaches, and meningeal irritation. One patient had papillitis and a maculopapular rash. Lymphadenopathy, which is a common finding in West Nile fever, was not found in any of our patients. Duration of the disease was one to two weeks, and recovery was complete. Cerebrospinal fluid examination revealed an increase in protein and pleocytosis (predominantly polymorphonuclears). We believe that West Nile encephalitis is not rare in Israel. PMID- 6274826 TI - [Detection of papilloma virus in Heck's focal epithelial hyperplasia and the differential diagnosis of white-sponge nevus]. AB - Clinical, histological, and electron microscopical findings helped in to differentiating hyperplasia of a Turkish boy's oral mucosa as Heck's disease, and the hyperplasia of a young German man's oral mucosa as white sponge nevus. Electron microscopic investigations confirmed the presence of papillomavirus in the lesions of FEH. PMID- 6274827 TI - A quantitative approach to trace the corticotrophs in culture after adrenalectomy. AB - Anterior pituitaries of adrenalectomized and sham operated adult rats were dispersed by trypsin and cultured for 4 and 8 days. Adrenalectomy caused a moderate increase in number of corticotrophs in both zero-time cell suspensions and cultures. There was a pronounced elevation of immunoreactive ACTH content in both cells and media and an enhanced secretory response to stimulation of cultures with stalk-median eminence extract containing cortiocotropin releasing (CRF) activity. Some cells identified as corticotrophs by a specific immunostaining incorporated tritiated thymidine into their nuclei suggesting their ability to enter the cell cycle. The relatively smaller increase in number of ACTH cells and the considerably higher ACTH producing capacity of the corticotrophs after adrenalectomy seem to be inconsistent with the quantal response model of hormone secretion recently introduced by Rodbard. PMID- 6274828 TI - Silicotungstic acid for cytochemical localization of water soluble substance(s) of cholinergic motor nerve terminal. AB - Silicotungstic acid (STA), an electron dense substance and a powerful precipitating agent of quaternary ammonium salts such as choline and acetylcholine, was employed on the frog motor end-plate in order to prove that STA reacts with diffusible substance(s) in nerve terminals. Thus, STA treatment and osmium tetroxide (OsO4) fixation were performed in three different ways. No reaction was detectable when STA treatment followed osmification, while simultaneous treatment with STA and OsO4 darkened both presynaptic and synaptic vesicle membranes. When STA was employed directly on fresh tissues which were subsequently fixed by OsO4, small black precipitates were observed in the synaptic vesicles and none on other synaptic structures. The possible reaction of STA with acetylcholine is discussed. PMID- 6274829 TI - Cytochemical detection of mannose-specific receptors for glycoproteins with horseradish peroxidase as a ligand. AB - Horseradish peroxidase (HRP), a glycoprotein rich in mannose groups, was used as a ligand to detect receptors for glycoproteins in formalin-fixed, frozen sections of rat liver. Specific binding of HRP occurred to surface membranes of sinusoidal cells but not to those of parenchymal cells. The binding sites were visualized after the peroxidatic reaction in erythrocytes had been suppressed by methanol H2O2 and phenylhydrazine, the latter reagent also decreasing the nonspecific background adsorption of HRP. Several factors influencing the reaction were studied systematically. The specific binding of HRP to sinusoidal cells was greatly decreased or abolished when tissue blocks were fixed for longer than 1-2 h in a cold 4% formaldehyde solution and the frozen sections subsequently treated for 30 min in cold methanol. The specific binding of HRP increased when the concentration of HRP in the medium was increased from 10 microgram/ml to 40 microgram/ml, when the time of incubation with HRP was increased from 1 h to 4 h, or when the temperature of incubation with HRP was increased from 4 degrees C to 22 degrees C, the pH of the incubation medium was increased from 7.0 to 10.0. Little or no specific binding of HRP was observed in the absence of added Ca++. The binding of HRP was suppressed by 10 mM mannose or 0.004% mannan whereas the suppression of the binding reaction by galactose or galactan required 30-40 ties higher concentrations. PMID- 6274830 TI - Comparative immunocytochemical localization of putative opioid ligands in the central nervous system. AB - We report a detailed comparative immunocytochemical mapping of enkephalin, CCK and ACTH/beta-endorphin immunoreactive nerves in the central nervous system of rat and guinea pig. Enkephalin immunoreactivity was detected in many groups of nerve cell bodies, fibers and terminals in the limbic system, basal ganglia, hypothalamus, thalamus, brain stem and spinal cord. beta-endorphin and ACTH immunoreactivity was limited to a single group of nerve cell bodies in and around the arcuate nucleus and in fibers and terminals in the midline areas of the hypothalamus, thalamus and mesencephalic periaqueductal gray with lateral extensions to the amygdaloid area. Cholecystokinin immunoreactive nerve fibers and terminals displayed a distribution similar to that of enkephalin in many regions; but striking differences were also found. An immunocytochemical doublestaining technique, which allowed simultaneous detection of two different peptides in the same tissue section, showed that enkephalin-, CCK- and ACTH/beta endorphin-immunoreactive nerves although closely intermingled in many brain areas, occurred separately. The distributions of nerve terminals containing these neuropeptides showed striking overlaps and also paralleled the distribution of opiate receptors. This may suggest that enkephalin, CCK, ACTH and beta-endorphin may interact with each other and with opiate receptors. PMID- 6274831 TI - [Intraarterial therapy of maxillary adenoid cystic carcinoma using cis-platinum (author's transl)]. AB - A woman suffering from extended maxillary adenoid cystic carcinoma of mixed cribriform solid type was treated by selective intraarterial infusion of Cisplatinum using a new intraarterial infusion technique. Within four weeks--the entire dosage of Cisplatinum was 400 mg--the patient came into complete remission, clinically and histologically, suggesting that the adenoid cystic carcinoma may be more susceptible to chemotherapy than thought up to the present day. However, primary chemotherapy should be followed by radiotherapy or surgical treatment. PMID- 6274832 TI - Effect of an exercise run to exhaustion on cAMP in the rat heart. AB - The purpose of this investigation was to determine whether adenosine 3',5'-cyclic monophosphate (cAMP) content is increased in vivo in the heart as a result of exercise at a time when there is rapid cardiac glycogen utilization. Rats were run to exhaustion on a treadmill for a period of 164.5 +/- 9.5 min. Blood norepinephrine and epinephrine were significantly elevated approximately 2.5-fold above resting levels at the end of the treadmill run. Myocardial glycogen was reduced by 54.7% at exhaustion compared with control values. Myocardial cAMP was significantly elevated 88% above control levels as a result of the run. Associated with the depletion of myocardial glycogen and the elevation of cAMP was an activation of phosphorylase to its a form. These data suggest that myocardial glycogen metabolism during exercise is, in part, mediated by hormonal influences that are associated with increases in cAMP. PMID- 6274833 TI - Ultrastructural changes in atrial myocardium of the ageing rat. AB - Ageing changes in the fine structure of atrial myocardial cells were studied in rats ranging from 2-32 months in age. The most striking change observed was the increasingly frequent appearance, from about 6 months onwards, of helically aggregated strands containing filaments which in respects other than arrangement bore resemblance to thick myofilaments. Other ageing changes included the accumulation of dense bodies and various types of mitochondrial degradation. PMID- 6274834 TI - Comparison of the renal excretory mechanisms of cefmenoxime and other cephalosporins: effect of para-aminohippurate on renal clearance and intrarenal distribution of cephalosporins in rabbits. AB - The renal excretory mechanism of cefmenoxime in rabbits was compared with that of 6 other cephalosporins (cefotaxime, deacetylcefotaxime, cefotiam, cefazolin, cephaloridine, and cefsulodin). The clearance ratios (Cf-Drug/CInulin=CRf) of cefmenoxime (337) and cefazolin (73) were considerably higher than those of the 5 other cephalosporins (0.9-20). When p-amino-hippurate (PAH) was administered concurrently with each of the cephalosporins, the CRf values of the cephalosporins except for cefsulodin were significantly decreased. These findings indicate that cefmenoxime and the 5 other cephalosporins except cefsulodin are actively incorporated in the proximal tubular cells and secreted into the tubular lumen. In the case of cefotiam and cefsulodin, glomerular filtration tended to exceed urinary excretion with highest dose of PAH (40 mg/kg/minute), suggesting the possibility of tubular reabsorption of these drugs. On the other hand, glomerular filtration of cefmenoxime and the 4 other cephalosporins did not exceed urinary excretion. The drug concentration ratio of the cortex to medulla indicated that the tubular cell level of cefmenoxime was lower than, higher than, and similar to those of cephaloridine, cefotaxime, and the remaining cephalosporins, respectively. These results demonstrate that the renal excretory mechanisms of cefmenoxime is similar to that of cefazolin but not to that of the remaining cephalosporins. PMID- 6274835 TI - Microbial resistances: a predictive test for streptothricin antibiotics. PMID- 6274836 TI - Light and electron microscopy of the morphological response of Escherichia coli and Serratia marcescens to cefmenoxime (SCE-1365), a new broad-spectrum cephalosporin. AB - The morphological response of Escherichia coli and Serratia marcescens to cefmenoxime (SCE-1365), 7 beta-[2-(2-aminothiazol-4-yl)-(Z)-2 methoxyminoacetamido]-3-[(1-methyl-1H-tetrazol-5-yl)thiomethyl]ceph-3-em-4 carboxylic acid, a new broad-spectrum cephalosporin, was investigated. The action of cefmenoxime was bactericidal against both E. coli and S. marcescens even at rather low concentrations. The pattern and the rate of the decrease of colony forming units (CFU) were similar over a fairly wide range of concentration; this was especially noticeable in S. marcescens. In E. coli filamentous cells were induced at low concentrations of the cephalosporin. Spheroplasts appeared at higher concentrations, and they lysed later. A bulge was formed at the middle of the cell at concentrations near the minimal inhibitory concentration (MIC), and vacuole-like structures surrounded by membrane were also observed in the cytoplasm of the filamentous cells. In S. marcescens filamentation of cells occurred over a considerable range of concentration. With longer times of incubation, granular structures and fused nuclear regions were noticed in the sparse cytoplasm. PMID- 6274837 TI - Comparison of the renal excretory mechanisms of cefmenoxime and other cephalosporins: renal clearance in rats and rabbits. AB - The renal excretory mechanism of cefmenoxime was compared with those of 6 other cephalosporins (cefotaxime, deacetylcefotaxime, cefotiam, cefazolin, cephaloridine and cefsulodin) in rats and rabbits. In rats, renal clearance (Cf Drug: corrected for serum protein binding) of cefmenoxime (4.06 ml/minute) was lower than that of cefazolin, similar to that of cefotiam, and higher than those of cefotaxime, deacetylcefotaxime, cephaloridine, and cefsulodin. In rabbits, the value for cefmenoxime (2,519 ml/minute) was markedly higher than those for the other drugs. If the clearance ratio (Cf-Drug/CInulin = CRf) is more than unity, tubular secretion of a drug is indicated. In rats, the tubular secretion of cefmenoxime equalled the glomerular filtration (CRf = 2.17). The tubular secretion of cefmenoxime was lower than that of cefazolin but similar to that of cefotiam. None of the other cephalosporins seemed to be secreted significantly. In rabbits, a large amount of cefmenoxime was secreted (CRf = 208). The tubular secretion was larger than those of the other cephalosporins. All the drugs except cefsulodin showed significant tubular secretion. A species difference was observed in the CRf of the cephalosporins: the values were generally higher in rabbits than in rats. PMID- 6274838 TI - Preovulatory changes prostaglandins E2 and F2 alpha in ovine follicles. PMID- 6274839 TI - Neurological complications of spinal analgesia. PMID- 6274840 TI - Insertion of transposons through the major cotransduction gap of Escherichia coli K-12. AB - The major cotransduction gap of the Escherichia coli chromosome extends from mini 31 to 34. We have inserted transposons through this gap which, by sequential transduction, link sbcA (min 29.8) with manA (min 35.7) and thus eliminate the gap. These results indicate that the length of DNA in the region, as measured by transduction, is not significantly different from the length obtained by conjugational time of entry. Since this segment of the E. coli chromosome has few known genes, these transposon insertions will be useful for genetic manipulations in the region of the gap. We describe the usefulness of these markers for rapidly mapping mutations which may be isolated in the region from min 27 to 37. PMID- 6274841 TI - Physical and genetic characterization of symbiotic and auxotrophic mutants of Rhizobium meliloti induced by transposon Tn5 mutagenesis. AB - We have physically and genetically characterized 20 symbiotic and 20 auxotrophic mutants of Rhizobium meliloti, the nitrogen-fixing symbiont of alfalfa (Medicago sativa), isolated by transposon Tn5 mutagenesis. A "suicide plasmid" mutagenesis procedure was used to generate TN-5-induced mutants, and both auxotrophic and symbiotic mutants were found at a frequency of 0.3% among strains containing random TN5 insertions. Two classes of symbiotic mutants were isolated: 4 of the 20 formed no nodules at all (Nod-), and 16 formed nodules which failed to fix nitrogen (Fix-). We used a combination of physical and genetic criteria to determine that in most cases the auxotrophic and symbiotic phenotypes could be correlated with the insertion of a single Tn5 elements. Once the Tn5 element was inserted into the R. meliloti genome, the frequency of its transposition to a new site was approximately 10-8 and the frequency of precise excision was less than 10-9. In approximately 25% of the mutant strains, phage Mu DNA sequences, which originated from the suicide plasmid used to generate the Tn5 transpositions, were also found in the R. meliloti genome contiguous with Tn5. These later strains exhibited anomalous conjugation properties, and therefore we could not correlate the symbiotic phenotype with a Tn5 insertion. In general, we found that both physical and genetic tests were required to fully characterize transposon-induced mutations. PMID- 6274842 TI - Beta-lactamase-specifying plasmids isolated from Neisseria gonorrhoeae have retained an intact right part of a Tn3-like transposon. AB - In three beta-lactamase-producing strains of Neisseria gonorrhoeae, ampicillin resistance is due to the presence of a 7.4-kilobase plasmid. Heteroduplex analysis has shown that the R plasmid contains a 1.6-kilobase segment homologous to the right part (the region coding for the beta-lactamase) of the Tn3-like transposon Tn2301 the 1.6-kilobase DNA segment is not transposable, but it can give rise to a functional transposon, when linked to the left part of TN2301. This provides strong evidence that the R plasmids of N. gonorrhoeae are deletion derivatives of a plasmid that contained an entire TN3-like transposon. PMID- 6274843 TI - Effect of catabolite repression on the mer operon. AB - The plasmid-determined mer operon, which provides resistance to inorganic mercury compounds, was subject to a 2.5-fold decrease in expression when glucose was administered at the same time as the inducer HgCl2. This glucose-mediated transient repression of the operon was overcome by the addition of cyclic AMP. Permanent catabolite repression of the operon was observed in the 1.6- to 1.9 fold decrease in expression in mutants lacking either adenyl cyclase (cya) or the catabolite activator protein (crp). The effect of the cya mutation on mer expression could be overcome by the addition of cyclic AMP at the time of induction, In addition to these effects on the whole cells of a wild-type strains, we examined the effect of catabolite repression on the expression of the mercuric ion [Hg(II)] reductase enzyme, assayable in cell extracts, and on the Hg(II) uptake system, assayable in a mutant strain which lacked reductase activity. There was a two- to threefold effect of repression on the Hg(II) reductase enzyme assayable in vitro after induction under catabolite repressing conditions (either with glucose or in the crp and cya mutants). We did not find a similar repressing effect on the induction of the Hg(II) uptake system, which is also determined by the mer operon. PMID- 6274844 TI - Clustering of nitrogen fixation (nif) genes in Rhizobium meliloti. AB - A cloned 17.3-kilobase region of the Rhizobium meliloti genome with homology to the Klebsiella pneumoniae nitrogenase structural genes was studied. Limits on the extent of homology were determined. Transposon mutagenesis of this region of the genome verified that it contained functional nif genes, Some transposon insertions resulted in a defective symbiotic phenotype, whereas others had no noticeable effect on symbiotic competence. The relative position of insertions yielding these two phenotypic classes suggested that at least three distinct units of gene expression are present in this region. Hybridization of RNA from alfalfa root nodules and from vegetatively grown Rhizobium to this cloned DNA showed that at least 11.1 kilobases of the region was transcribed actively and that transcription was specific for the symbiotic state. PMID- 6274845 TI - Mapping of a plasmid, coding for colonization, factor antigen I and heat-stable enterotoxin production, isolated from an enterotoxigenic strain of Escherichia coli. AB - The non-autotransferring plasmid NTP113 codes for production of colonization factor antigen I and heat-stable enterotoxin, NTP113, which has a molecular weight of 58 X 10(6), was digested with BamHI, EcoRI, and HindIII and combinations of these restriction endonucleases, and the products of these digestions were analyzed by agarose gel electrophoresis. The results were used to construct a partial restriction map of NTP113. Transposons coding for resistance to ampicillin, kanamycin, and tetracycline were inserted into NTP113, and we obtained a series of deletion mutants, as determined by the loss of tetracycline or kanamycin resistance from strains carrying the insertion mutants. A number of plasmid mutants obtained by insertion or deletion did not code for colonization factor antigen I, but most of these mutants still coded for heat-stable enterotoxin production. The position of the inserted transposons and of the deletions were determined on the restriction map. Two regions of NTP113 were required for the expression of colonization factor antigen I, and the two sites were separated by a length of DNA corresponding to a molecular weight of about 25 X10(6). PMID- 6274846 TI - Physical and genetic analysis of deletion mutants of plasmid R91-5 and the cloning of transfer genes in Pseudomonas aeruginosa. AB - We isolated deletion mutants of Pseudomonas aeruginosa plasmid R91-5 by both in vitro and in vivo means. Many of the deletion mutants selected on the basis of resistance to donor-specific phages fell into a few groups of apparently identical mutants, although the mutants were nonsibs. By analyzing plasmids with large deletions, we found that the essential replication genes of R91-5 were within a 3.85-kilobase region between coordinates 45.5 and 48.9. The origin of plasmid transfer (oriT) was mapped to a 4.5-kilobase region between coordinates 1.7 and 6.2. We indirectly determined the direction of plasmid transfer from oriT. By combining the data from our analysis of the deletions with data from complementation tests between cloned R91-5 fragments and known reference mutants, we ordered and mapped the 10 known transfer (tra) cistrons of R91-5. All of the tra cistrons mapped within the Tra2 region, and their order was as follows: traX, -Y, -T, -Q, -(V, R), -U, -(S, Z), -W (the cistrons in parentheses could not be ordered with respect to each other). PMID- 6274847 TI - Cloning characterization of iaaM, a virulence determinant of Pseudomonas savastanoi. AB - Genes for indoleacetic acid production (iaaM and iaaH) are necessary for gall induction by the olive pathogen Pseudomonas savastanoi. In strain 2009 these determinants are borne on plasmid pIAA1. To map and characterize the genes, fragments of pIAA1 generated by EcoRI endonuclease treatment were cloned in Escherichia coli by using plasmid RSF1010 as vector. We isolated a recombinant plasmid encoding iaaM, the locus for tryptophan 2-monooxygenase. This plasmid, called pLUC1, was characterized by restriction endonuclease hydrolysis. It contained a 2.75-kilobase-pair segment of pIAA1. By cloning this segment in the EcoRI site of pBR328 and pBRH3B we showed that efficient expression of iaaM was dependent on the orientation with respect to the vector promoters, and thus determined the direction of transcription. To more finely map iaaM and confirm the orientation of transcription, plasmid pLUC1 was subjected to transposon Tn/mutagenesis. The promoter-distal end of iaaM was mapped between coordinates at 1.7 and 2.15 kilobase pairs of the cloned segment. PMID- 6274848 TI - Amino-terminal sequence of the Tn3 transposase protein. AB - The amino-terminal sequence of the Tn3 transposase protein was determined to be Pro-Val-Asp-Phe-Leu-Thr-Thr-Glu-Gln-Val-Glu-Ser.... This was determined both from an active transposase protein purified from a transposase overproducing mutant strain and from a hybrid transposase-beta-galactosidase fusion protein. The amino acid sequence corresponded to the DNA sequence of the transposase gene beginning at an ATG initiation codon, as previously predicted from the analysis of transposase-beta-galactosidase gene fusions. PMID- 6274850 TI - Analogs of cyclic AMP as chemoattractants and inhibitors of Dictyostelium chemotaxis. AB - Aggregative amoebae of Dictyostelium discoideum, D. mucoroides, D. purpureum, and D. rosarium react chemotactically to cyclic AMP (cAMP). We measured the chemotactic activity of 14 cAMP analogs and found that these four species have a similar sensitivity to chemical modifications of cAMP; this suggests that the cAMP receptor is identical in all of these species. Besides the induction of a chemotactic response, cAMP analogs also may delay or prevent cell aggregation. cAMP analogs like N1-O-cAMP, 2'-H-cAMP, and 5'NH-cAMP are chemotactically nearly as active as cAMP and induced no, or only a short, delay of cell aggregation. Other cAMP derivatives, such as 6-Cl-cPMP and 8-Br-cAMP, are chemotactically active only at high concentrations and delayed cell aggregation for several hours. Still other cAMP analogs, which do not induce a chemotactic reaction in D. mucoroides, D. purpureum, and D. rosarium, either prevented cell aggregation [cAMPS(S), cAMPS(R), and 3'-NH-cAMP[ or had no effect on cell aggregation [cAMPN(CH3)2(S) and cAMPN(CH3)2(R)]. cAMP analog 3'-NH-cAMP prevented cell aggregation by the inhibition of chemotaxis, whereas cell locomotion was not affected. Although we cannot provide a satisfactory explantation for these observations, our data suggest that occupation and activation of the cAMP receptors do not always induced a chemotactic response. PMID- 6274849 TI - Differential effects of antibiotics inhibiting gyrase. AB - Both oxolinic acid and coumermycin A1, inhibitors of DNA gyrase, block DNA synthesis in Escherichia coli. At low concentrations of oxolinic acid, the rate of bacterial DNA synthesis first declines rapidly but then gradually increases. This gradual increase in synthesis rate depended on the presence of wild-type recA and lexA genes; mutations in either gene blocked the increase in synthesis rate. In such mutants, oxolinic acid caused a rapid decline, followed by a slow, further decrease in DNA synthesis rate. Coumermycin A1, however, produced a more gradual decline in synthesis rate which is unaffected by defects in the recA or lexA genes. An additional difference between the two drugs was observed in a dnaA mutant, in which initiation of replication is temperature sensitive. Low concentrations of oxolinic acid, but not coumermycin A1, reduced thermal inhibition of DNA synthesis rate. PMID- 6274851 TI - A homogeneous cyclic CMP phosphodiesterase hydrolyzes both pyrimidine and purine cyclic 2':3'- and 3':5'-nucleotides. AB - A homogeneous preparation of cyclic CMP phosphodiesterase (Helfman, D. M., Shoji, M., and Kuo, J. F. (1981) J. Biol. Chem. 256, 6327-6334) was found to catalyze the hydrolysis of both pyrimidine and purine cyclic 2':3'- and 3':5'-nucleotides. Hydrolysis of cyclic 2':3'-nucleotides resulted in the formation of both 2'- and 3'-nucleotides, although relative amounts of the products were variable. Hydrolysis of cyclic 2':3'-CMP or cyclic 2':3'-UMP yielded predominantly 3' nucleotides. In contrast, hydrolysis of cyclic 2':3'-AMP produced equal amounts of 2'- and 3'-nucleotides, while the major product formed from cyclic 2':3'-GMP was 2'-nucleotide. When conventional pyrimidine and purine cyclic 3:5' nucleotides were used as substrates, the enzyme hydrolyzed specifically the 3' bond to yield only 5'-nucleotides. The relative rate of hydrolysis of cyclic 2':3'-nucleotides was cyclic CMP greater than cyclic UMP greater than cyclic GMP greater than or equal to cyclic AMP, respectively, whereas that for cyclic 3':5' nucleotides was cyclic CMP greater than cyclic UMP greater than or equal to cyclic AMP greater than cyclic GMP, respectively. Furthermore, kinetic analysis suggested a single species of catalytic site on the enzyme may be involved in the hydrolysis of both pyrimidine and purine cyclic 2':3'- and 3':5'-nucleotides. These findings indicate that the present enzyme is the first multifunctional phosphodiesterase reported to date that is capable of hydrolyzing such a diversity of cyclic nucleotides. PMID- 6274852 TI - The sequence of a mouse embryonic beta-globin gene. Evolution of the gene and its signal region. AB - We have determined the nucleotide sequence of an embryonic beta-globin gene of the Balb/c mouse. It possesses structural features typical of known expressed beta-globins, including 5'-untranslated region, potential capping site, initiation and terminator codons, poly(A) addition signal, splicing signals, and intervening sequences. There is a striking 15-base homology between the putative RNA polymerase binding site of this gene and the corresponding region of a human embryonic beta-gene which may be important in the control of expression during embryonic development. The sequence of the gene predicts the sequence of the entire y2 protein, which has not previously been available. As expected from evolutionary considerations, it is more homologous to human embryonic beta-globin than to mouse adult beta-globin. PMID- 6274853 TI - Nucleotide sequences of transcription and translation initiation regions in Bacillus phage phi 29 early genes. AB - phi 29 DNA directs the synthesis of three major proteins of Mr = 22,400, 13,900, and 10,500 in a cell-free transcription-translation system derived from Bacillus subtilis. We have determined the locations of the coding regions for these early proteins on the phi 29 genome, and our results are in agreement with genetic evidence that the 22.4-kilodalton protein is the product of cistron 17 (p17) and the 13.9-kilodalton protein is the product of cistron 6 (p6). The 13.9-kilodalton and 10.5-kilodalton proteins are encoded on a polycistronic mRNA previously designated G3b RNA. We have determined the nucleotide sequence of a HindIII restriction fragment of phi 29 DNA, the H fragment, that encodes the 13.9 kilodalton protein and contains two early promoters, G3a and G3b. The nucleotide sequence of the ribosome binding site contains a polypurine region capable of forming a very stable complex with nine bases on the 3' end of B. subtilis 16 S rRNA. This strong Shine-Dalgarno complementarity supports the hypothesis that an extensive mRNA . rRNA interaction is a requirement for efficient translation by B. subtilis ribosomes (McLaughlin, J. R., Murray, C. L., and Rabinowitz, J. C. (1981) J. Biol. Chem. 256, 11283-11291). The nucleotide sequences of both promoters are very similar to the "consensus" sequence for Escherichia coli promoters. While E. coli RNA polymerase initiates transcription from both of these promoters in vitro, only the G3b promoter is utilized by B. subtilis RNA polymerase under the same conditions. PMID- 6274854 TI - Biosynthesis of galactosyl-beta 1,3-N-acetylglucosamine. AB - The biosynthesis of galactosyl-beta 1,3-N-acetylglucosamine has been demonstrated using membrane preparations from pig trachea. Unlike the UDP-galactose:2 acetamido-2-deoxy-D-glucose 4 beta-galactosyltransferase, which is inhibited by high levels of N-acetylglucosamine, the UDP-galactose:N-acetylglucosamine 3 beta galactosyltransferase shows no inhibition at 200 mM N-acetylglucosamine. About 80% of the total disaccharide synthesized at 200 mM N-acetylglucosamine was base labile suggesting the 1,3-linkage, alpha-Lactalbumin inhibits galactose incorporation into galactosyl-beta 1,4-N-acetylglucosamine but has little or no effect on the activity of the 1,3-galactosyltransferase. Escherichia coli beta galactosidase readily hydrolyzed the base-stable product, but not the base-labile component. The apparent 1,3-linked disaccharide was reduced with NaBH4 and was isolated by Bio-Gel P-2 column chromatography. Methylation analysis by gas chromatography/mass spectrometry showed tetramethyl galactose and a 3-substituted N-acetylglucosaminitol. Neither the beta 1,4 nor the beta 1,3 disaccharide was hydrolyzed by green coffee bean alpha-galactosidase. Both disaccharides were readily hydrolyzed by bovine testes beta-galactosidase. This is the first report on the galactosyltransferase which catalyzes the synthesis of the galactosyl-beta 1,3-N-acetylglucosamine linkage such as found in the Type I chain of human blood group substances. A tissue survey in rats showed only rat intestine to have readily detectable UDP-galactose: N-acetylglucosamine 3 beta galactosyltransferase activity. The intestinal membrane fraction like the tracheal enzyme catalyzes the synthesis of two disaccharides as judged by base treatment, and these appear to be the beta 1,3 and beta 1,4 isomers of galactosyl N-acetylglucosamine. PMID- 6274855 TI - Incorporation of E-5-(2-halovinyl)-2'-deoxyuridines into deoxyribonucleic acids of herpes simplex virus type 1-infected cells. AB - E-5-(2-Bromovinyl)-2'-deoxyuridine (BrvdUrd) and E-5-(2-iodovinyl)-2' deoxyuridine (IvdUrd) are among the most potent and selective inhibitors of herpes simplex virus type 1 (HSV-1) replication. To elucidate the site of inhibition, we examined whether the halovinyl analogs are incorporated into DNA using two approaches. (i) In assays with purified DNA polymerases omitting dTTP from the reaction system, addition of either BrvdUTP or IvdUTP increased the polymerization reaction, indicating that these two analog triphosphates can be alternate substrates. (ii) When HSV-1-infected Vero cells were grown in the presence of either BrvdUrd or IvdUrd, there was an increase in the density of both the viral and cellular DNA. The viral DNA had 40% of its thymidine moiety substituted by IvdUrd when the concentration of [125I]IvdUrd was 24 microM (in the absence of added thymidine). At 30 microM BrvdUrd and 1 microM [2 14C]thymidine, the viral DNA had only 11% of its thymidine moiety substituted by BrvdUrd, presumably because of the presence of added thymidine. Following digestion of [125I]IvdUrd-substituted DNA with DNase 1, venom phosphodiesterase, and alkaline phosphatase, the radioactivity co-migrated with nonradioactive IvdUrd in thin layer chromatography. Under similar conditions, no detectable incorporation of either [125I]IvdUrd or BrvdUrd into mock-infected Vero cell DNA was observed. Thus, IvdUrd and BrvdUrd are incorporated into DNA of HSV-1 infected cells but not into DNA of uninfected cells. PMID- 6274856 TI - Poly(ADP-ribose) has a branched structure in vivo. AB - We have searched for the presence of branching in the chromosomal polymer poly(ADP-ribose) as it occurs in vivo. Treatment of the polymer with phosphodiesterase asnd phosphomonoesterase results in the conversion of internal residues to the nucleoside ribosyladenosine and the conversion of points of branching to diribosyladenosine. We have detected diribosyladenosine in digests of the polymer derived from carcinogen-treated SV40 virus-formed 3T3 cells and in normal rat liver, kidney, and spleen. The frequency of residues involved in branching varied from 0.8 to 1.6 mole % over a 50-fold range of total levels of poly(ADP-ribose). Thus, branching seems to be a general feature of poly(ADP ribose) as it occurs in vivo. PMID- 6274857 TI - Characterization of a follitropin-binding component prepared from immature bovine testes in the absence of detergent. PMID- 6274858 TI - Demonstration of receptors for insulin and insulin-like growth factors on Swarm rat chondrosarcoma chondrocytes. Evidence that insulin stimulates proteoglycan synthesis through the insulin receptor. AB - The insulin-like growth factor, multiplication stimulating activity (MSA), and insulin were recently shown to stimulate proteoglycan synthesis in monolayer cultures of chondrocytes derived from the Swarm rat chondrosarcoma. Insulin produced significant stimulation at a concentration of less than 1 ng/ml, suggesting that insulin was acting through the insulin receptor rather than through a somatomedin receptor. In this paper we have shown that the insulin-like growth factors IGF-I and IGF-II also are potent stimulators of [35S]sulfate incorporation into macromolecules recovered from the medium and cell layer matrix of the chondrosarcoma chondrocytes. Proinsulin was 3% as potent as insulin in stimulating [35S]sulfate incorporation. We identified receptors for insulin and the insulin-like growth factors, MSA, IGF-I, and IGF-II. Insulin, at concentrations 1000 times the concentration required to produce the biologic response, did not compete for binding of 125I-MSA-II-1 or of 125I-IGF-II and only partially competed for 125I-IGF-I binding. Anti-insulin receptor IgG stimulated proteoglycan synthesis and competed for 125I-insulin binding. Fab fragment prepared from anti-insulin receptor IgG completely blocked the stimulation of [35S]sulfate incorporation into macromolecules by insulin while only partially inhibiting the biologic response to insulin-like growth factors, MSA, IGF-I, and IGF-II. Similarly, the anti-insulin receptor IgG only partially inhibited the binding of 125I-IGF-I and 125I-IGF-II while completely blocking the binding of 125I-insulin. We conclude that insulin stimulates proteoglycan synthesis in the chondrosarcoma chondrocytes by acting through the insulin receptor whereas the insulin-like growth factors probably act through their own receptors. PMID- 6274859 TI - Nucleotide sequence of bovine prolactin messenger RNA. Evidence for sequence polymorphism. AB - Hybrid molecules containing DNA sequences complementary to bovine pituitary mRNA were constructed in the Pst I site of pBR322 by the dC . dG tailing technique. Recombinant plasmids containing bovine prolactin (bPRL) sequences were amplified in bacteria and identified by hybridization to purified [32P]bPRL cDNA sequences. Nucleotide sequence analysis was performed on the inserts from two of the positive clones. One clone, pBPRL72, contained a 982-base pair insert that included 67 nucleotides of the 5'-untranslated region, the complete coding region of the preprolactin protein (690 nucleotides), and the entire 3'-untranslated region (150 nucleotides) of bPRL mRNA. The nucleotide sequence analysis of clone pBPRL72 predicted the sequence of a 30-amino acid signal peptide and confirmed the published amino acid sequence of the protein with one exception. A comparison of the pBPRL72 cDNA sequence with a second bPRL clone, pBPRL4, revealed four silent nucleotide differences. Three of the base changes occurred in the third position of amino acid codons, and one occurred in the 3'-noncoding region. The sequence polymorphism suggests the existence of alleles or multiple loci for bPRL that do not alter the protein structure. PMID- 6274860 TI - The size of the thyroid hormone receptor in chromatin. AB - We have used radiation inactivation and target theory to determine the size of the functional unit for T3 binding in rat liver chromatin. The process involves exposure of frozen chromatin samples to a beam of high energy electrons produced in a linear accelerator and subsequent measurement of the residual capacity to bind hormone. Our experiments were carried out using three forms of solubilized chromatin: 1) sonicated, containing the receptor in fragments which sedimented faster than 30 S; 2) digested by nuclease, containing the receptor in a form which sedimented at 5-6 S; 3) digested by nuclease and made 0.5 M in KCl, containing the receptor in a form which sedimented at 3.8 S. We have shown that in each sample preparation the receptor retained the ability to bind T3 with the same capacity and affinity that had previously been measured with high molecular weight chromatin. Irradiation caused a reduction in the capacity to bind T3 but did not change the affinity of the remaining receptors for the hormone. In each preparation, the radiation resulted in a simple exponential loss of binding capacity with dose, indicating that a single target size was detected. Within the variation of the measurements, the target size for each form of the receptor was the same, 59,000 daltons. PMID- 6274861 TI - Differences in hyaluronate binding to plasma and cell surface fibronectins. Requirement for aggregation. PMID- 6274862 TI - Amino acid sequence at the ATP-binding site of cGMP-dependent protein kinase. AB - The amino acid sequence at the ATP-binding site on the cGMP-dependent protein kinase has been determined. For this determination the enzyme was labeled covalently by 5'-p-fluorosulfonyl[14C]benzoyladenosine and fragmented using cyanogen bromide or digested by trypsin after succinylation. The 14C-labeled peptides were purified by gel filtration and high performance liquid chromatography. The amino acid sequence around the site was found to be: -Val-Glu Leu-Val-Gln-Leu-Lys-Ser-Glu-Glu-Ser-Lys-Thr-Phe-Ala-Met-*Lys-Ile-Leu-Lys--Lys-Arg His-Ile-Val-Asp-Thr-Arg-Gln-Gln-Glu-His-Ile-Arg-Ser-Glu-Lys-, in which *Lys is the lysine residue that was modified by the affinity reagent. When this sequence was compared with that of the ATP-binding site of the catalytic subunit of cAMP dependent protein kinase, a high degree of structural homology was observed for this site in the two proteins. PMID- 6274863 TI - Studies of Bungarus fasciatus venom NAD glycohydrolase. Inhibition, modification, and transglycosidation reactions. AB - The properties of the NAD-binding domain of Bungarus fasciatus venom NAD glycohydrolase were investigated by a number of biochemical techniques. Inhibitor studies indicated that an intact dinucleotide was required for effective enzyme recognition. Adenosine derivatives, AMP, ADP, ATP, adenosine diphosphoribose (ADP rib), and 2'-phosphoadenosine diphosphoribose (phospho-ADP-rib), were linear competitive inhibitors of the NADase-catalyzed reaction. Nicotinamide analogs were noncompetitive inhibitors with Ki values that varied with the nature and position of substituent groups. Selective enzyme inactivation by 2,4-pentanedione indicated the importance of a lysinyl residue for NADase activity. The importance of a carboxyl group was also indicated by the sensitivity of the NADase toward N ethyl-5-phenyl-isoxazolium-3'-sulfonate (Woodward's Reagent K). Reagents which modified sulfhydryl, histidyl, and arginyl residues were not effective in inactivating B. fasciatus venom NADase. The purified snake venom NADase catalyzed a transglycosidation (pyridine base exchange) reaction. The functioning of a variety of substituted pyridine bases in this reaction was demonstrated by the formation of the corresponding NAD analogs. Kinetic studies of the transglycosidation reaction were consistent with the partitioning of an enzyme ADP-rib intermediate between water and the substituted pyridine base. Concentrations of pyridine bases required for enzyme inhibition differed significantly from those needed for the pyridine base exchange reaction, suggesting two sites or two forms of the NADase differing in affinity for pyridine bases. A number of NAD analogs were also shown to be effective ribosyl donors in the transglycosidation reaction. PMID- 6274864 TI - Effects of magnesium and N-ethylmaleimide on the binding of 3H hydroxybenzylisoproterenol to beta-adrenergic receptors. PMID- 6274865 TI - Irreversible inhibition of bovine lung angiotensin I-converting enzyme with p [N,N-bis(chloroethyl)amino]phenylbutyric acid (chlorambucil) and chlorambucyl L proline and with evidence that an active site carboxyl group is labeled. AB - Bovine lung angiotensin I-converting enzyme is rapidly and irreversibly inactivated by p-[N,N-bis(chloroethyl)amino]phenylbutyric acid (chlorambucil) and by the chlorambucil derivative of L-proline (chlorambucyl-proline). Chlorambucil is a nitrogen mustard alkylating agent that is used as an antineoplastic drug. At any one concentration, the inactivation is pseudo-first order with time. Inhibition by both substances is active site directed as suggested by the formation of a reversible enzyme-inhibitor complex prior to the alkylation reaction and by the fact that L-Phe-L-Pro, a reversible inhibitor which is competitive with substrate, is also competitive with both irreversible inhibitors in protecting the enzyme against inactivation. The second order rate constant for inactivation increases in the pH range 5-8 and reaches a value of 3.5 X 10(3) M-1 . min-1 for chlorambucil and 4.8 X 10(2) M-1 . min-1 for chlorambucyl-proline. Chlorambucyl [U-14C]L-proline reacts 1:1 with the converting enzyme and the uptake of radioactivity paralleled the loss of enzyme activity with and without protection by Phe-Pro. Once bound, the radioactive chlorambucyl proline was released (as the dihydroxy derivative) by hydroxide ion with a second order rate constant of 2.2 M-1 . min-1 at 25 degrees C. The radioactive label is also removed by hydroxylamine at pH 10. The lability of the irreversibly bound inhibitor in alkali and in hydroxylamine indicates that an ester bond is formed by the alkylation of an aspartic acid or glutamic acid side chain. PMID- 6274866 TI - Characterization of endogenous protein phosphorylation in isolated rat liver lysosomes. AB - Membranes prepared from highly purified rat liver lysosomes contain endogenous protein-phosphorylation activities. The transfer of phosphate to membrane fractions from [gamma-32P]ATP was analyzed by gel electrophoresis under acidic denaturing conditions. Two phosphopeptides were detected, with molecular weights of 3,000 and 14,000. Phosphorylation of these proteins was unaffected by the addition of cAMP, cGMP, or the heat-stable inhibitor of cAMP-dependent protein kinase. No additional phosphorylation was observed when cAMP-dependent protein kinase was included in the reaction or when exogenous protein kinase substrates were added. The 14,000-dalton 32P-labeled product was formed rapidly in the presence of low concentrations (250 microM) of either Ca2+ or Mg2+. This product was labile under both acidic and alkaline conditions, suggesting that this protein contains an acyl phosphate, present presumably as a catalytic intermediate in a phosphotransferase reaction. The lower molecular weight species required a high concentration (5 mM) of Mg2+ for phosphorylation, and micromolar concentrations of Ca2+ stimulated the Mg2+-dependent activity. The addition of Ca2+ and calmodulin stimulated the phosphorylation reaction to a greater extent than with Ca2+ alone. This activity was strongly inhibited by 0.2 mM LaCl3 and to a lesser extent by 50 microM chlorpromazine or trifluoperazine. These results suggest that the 3000-dalton peptide may be phosphorylated by a Ca2+, calmodulin dependent kinase associated with the lysosomal membrane. PMID- 6274867 TI - Incorporation of canavanine into rat pars intermedia proteins inhibits the maturation of pro-opiomelanocortin, the common precursor to adrenocorticotropin and beta-lipotropin. AB - Pro-opiomelanocortin, the common glycoprotein precursor to adrenocorticotropin and beta-lipotropin, is the most abundant protein synthesized in rat neurointermediate lobes. Dissected rat neurointermediate lobes were incubated in the presence of canavanine, an analog of arginine, to determine (a) whether canavanine could be incorporated into pro-opiomelanocortin molecules and (b) if incorporation occurs, whether there is any effect on the processing mechanism of the prohormone. Preincubation of rat neurointermediate lobes for 16 h in the presence of 10 mM canavanine results in the production of pro-opiomelanocortin molecules in which most, if not all, the arginine residues have been replaced by canavanine. Identification of canavanine-containing pro-opiomelanocortin forms was done by two-dimensional electrophoresis, tryptic and chymotryptic peptide mapping, as well as by analysis, on polyacrylamide gels in the presence of sodium dodecyl sulfate, of the fragments resulting from a partial digestion with chymotrypsin. During pulse-chase experiments, canavanine-containing pro opiomelanocortin molecules were found to be processed at a much slower rate than the normal precursor forms: after a 2-h chase, conversion of approximately 25% of the analog-containing prophormone was observed compared to 83% of the nonanalog containing precursors. Moreover, the small proportion of canavanine-containing precursor molecules which had undergone cleavage during the chase yielded atypical large molecular weight peptides. These results indicate that canavanine incorporation into neurointermediate lobe proteins considerably slows down the conversion of pro-opiomelanocortin into its different end products. PMID- 6274868 TI - A novel method for measuring cell surface-bound thrombin. Detection of iodination induced changes in thrombin-binding affinity. PMID- 6274869 TI - Characteristics and metabolism of alpha 1 adrenergic receptors in a nonfusing muscle cell line. AB - The BC3H1 nonfusing muscle cell line possesses binding sites for [3H]prazosin. These binding sites are typically alpha 1 adrenergic receptors as shown by their greater affinity (3700-fold) for prazosin than for yohimbine. Both kinetic and equilibrium analyses indicated that [3H]prazosin interacted with only one category of independent binding sites with the following characteristics. KD = 0.13 +/- 0.01 nM. Bmax = 97 +/- 5 fmol/mg of protein corresponding to 25,000 sites/cell (n = 17). Biosynthesis of the alpha 1 adrenergic receptor was investigated at cell confluency (when the number of cells and their total protein content were constant). Phenoxybenzamine (10(-9) M) irreversibly blocked 50% of the alpha 1 receptors in intact cells. More than 95% blockade of receptors was obtained with 10(-7) M phenoxybenzamine. After this blockade, new alpha 1 adrenergic receptors reappeared in the cells with monoexponential kinetics. These new receptors corresponded to synthesized receptors since their appearance was blocked by cycloheximide (1 micrograms/ml). The cycloheximide action was reversible. If one makes the simple and probable hypotheses that the receptor production is constant and that degradation is a monoexponential process, the analysis of the kinetics of reappearance allows the determination of the rate constant for receptor degradation (k = 0.03 h-1) and the rate of receptor production (r = 3.2 fmol/mg/h) corresponding to the synthesis of about 760 receptors/cell/h. The half-life of the receptor was 23 h. PMID- 6274870 TI - Effect of retinoic acid treatment of F9 embryonal carcinoma cells on the activity and distribution of cyclic AMP-dependent protein kinase. PMID- 6274871 TI - Creatine kinase of heart mitochondria. Functional coupling of ADP transfer to the adenine nucleotide translocase. PMID- 6274872 TI - Thyroid hormone nuclear receptor. Evidence for multimeric organization in chromatin. PMID- 6274873 TI - Translational control by messenger RNA competition for eukaryotic initiation factor 2. AB - Translation of globin mRNA in a micrococcal nuclease-treated reticulocyte lysate was studied in the presence of increasing amounts of Mengovirus RNA, under conditions in which the number of translation initiation events remains constant as judged by the transfer of label from N-formyl[35S]methionyl-tRNAf into protein. The translation of globin mRNA is progressively inhibited by low concentrations of Mengovirus RNA, free of detectable traces of double-stranded RNA, concomitant with the increasing synthesis of Mengovirus RNA-directed products. On a molar basis, Mengovirus RNA apparently competes about 35 times more effectively than globin mRNA for a critical component in translation. The competition is relieved by the addition of highly purified eukaryotic initiation factor 2 (eIF-2). Addition of eIF-2 does not stimulate overall protein synthesis, but shifts it in favor of globin synthesis. No stimulation of globin mRNA translation by eIF-2 is seen when Mengovirus RNA is absent. These experiments show that Mengovirus RNA competes, directly or indirectly, with globin mRNA for eIF-2. In direct binding experiments using isolated mRNA and eIF-2, Mengovirus RNA is shown to compete with globin mRNA for eIF-2 and to exhibit a 30-fold higher affinity for this factor. The binding of Mengovirus RNA to eIF-2 is much more resistant to increasing salt concentrations than is the binding of globin mRNA, again reflecting its high affinity. These results reveal a direct correlation between the ability of these mRNA species to compete in translation and their ability to bind to initiation factor eIF-2. They suggest that the affinity of a given mRNA species for eIF-2 is essential in determining its translation, relative to that of other mRNA species. Messenger RNA competition for eIF-2 may contribute significantly to the selective translation of viral RNA in infected cells. PMID- 6274874 TI - Cyclic AMP-dependent protein phosphorylation in canine renal brush-border membrane vesicles is associated with decreased phosphate transport. AB - It is known that the administration of parathyroid hormone to dogs results in phosphaturia and decreased phosphate transport in brush-border vesicles isolated from the kidneys of those dogs. Parathyroid hormone has been shown to activate adenylate cyclase at the basal-lateral membrane of the renal proximal tubular cell. It has been postulated that parathyroid hormone-induced phosphaturia is effected through phosphorylation of brush-border protein by membrane-bound cAMP dependent protein kinase. An experimental system was designed such that phosphorylation of brush-border vesicles and Na+-stimulated solute transport could be studied in the same preparations. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of membrane vesicles revealed cAMP-dependent phosphorylation of 2 protein bands (Mr = 96,000 and 62,000), which was enhanced by exposure of the inside of the membrane vesicles to ATP and cAMP. Cyclic AMP-dependent phosphorylation of brush-border vesicles was accompanied by inhibition of Na+ stimulated Pi but not D-glucose transport or 22Na+ uptake. When renal brush border vesicles from parathyroidectomized and normal dogs were phosphorylated in vitro in the presence and absence of cAMP, both the cAMP-dependent phosphorylation and inhibition of Na+-stimulated Pi transport were greater in vesicles isolated from kidneys of parathyroidectomized dogs relative to control animals. We conclude that the cAMP-dependent phosphorylation of brush-border membrane-vesicle proteins is associated with specific inhibition of Na+ stimulated Pi transport. The phosphaturic action of parathyroid hormone (PTH) could be mediated through the cAMP-dependent phosphorylation of specific brush border membrane proteins. PMID- 6274875 TI - Survival of bovine leukosis virus in bovine whole blood, serum and plasma. PMID- 6274876 TI - Pathological fracture of the radius through a cyst caused by pyrophosphate arthropathy. Report of a case. PMID- 6274877 TI - Synovial sarcoma. AB - One hundred and eighty-five patients, whose ages ranged from two to seventy years (mean, thirty years), have been treated at the Mayo Clinic for synovial sarcoma. Sixty-nine per cent of the tumors occurred in the lower extremity: 25 per cent, in the upper extremity; and 6 per cent, in the trunk. Histological characteristics included a predominant bimorphic pattern in 33 per cent of the tumors, a monomorphic pattern in 31 per cent, and a mixed pattern in 36 per cent. For all patients, the five-year survival rate was 38 per cent and the ten-year rate was 23 per cent, with a median survival time of thiry-nine months. However, for those patients treated since 1960, the five-year survival rate was 55 per cent and the ten-year rate was 38 per cent. Female patients had a significantly better survival time than did male patients. Younger patients had a better survival rate, and the prognosis was better for those patients whose tumor were either less than five centimeters in diameter or located in the lower extremity. Survival rates were best for those patients who had wide local excision, but this type of surgery was performed for the lesions that had a more favorable prognosis. A regression study indicated that the size of the tumor was the single most important prognostic variable for survival. Second in importance was the extent of disease on initial presentation, followed by the chronological time of treatment, the age of the patient, and the anatomical site of the tumor. PMID- 6274878 TI - The relationship between tumor promoter binding and Epstein-Barr virus induction in human lymphoblastoid cell lines. AB - The tumor promoter TPA (12-0-tetradecanoyl-phorbol-13-acetate) efficiently induces the synthesis of Epstein-Barr virus (EBV) antigens in EBV-genome harboring human lymphoblastoid cells. We attempted to obtain information on the binding of TPA to cells and on the relationship between TPA-binding and EBV induction by the use of tritiated TPA (3H-TPA). Our data show: (1) In the absence of cells TPA can bind to serum completely within 60 minutes. (2) Cells can compete for a proportion of serum-bound TPA. (3) Binding of TPA to cells reaches equilibrium within 60 minutes and is higher at 37 degrees than 0 degrees C. In the absence of serum, the rate of binding is about twice as high as in the presence of serum. (4) Dissociation of TPA from cells also seems to be rapid. When the cells are incubated with cold TPA after prior treatment with 3H-TPA, followed by washing, a much higher rate of release of labeled TPA is observed than in cultures to which fresh medium is added exclusively. Dissociation is higher in the presence of serum than in the absence of it. If radiolabeled cells are analyzed after serial washing, the rate of cell-associated/noncell-associated radioactivity indicates that the proportion of molecules required for EBV induction is dissociated rapidly. PMID- 6274880 TI - Alterations in calcitonin and parathyroid hormone responsiveness of adenylate cyclase in F9 embryonal carcinoma cells treated with retinoic acid and dibutyryl cyclic AMP. PMID- 6274879 TI - Functional receptors for epidermal growth factor on human osteosarcoma cells. PMID- 6274881 TI - Uptake of putrescine by 3T3 and SV3T3 cells and its effect on ornithine decarboxylase activity. PMID- 6274882 TI - Folate polyglutamate and monoglutamate accumulation in normal and SV40 transformed human fibroblasts. AB - Folate polyglutamate and monoglutamate accumulation was measured in normal diploid and SV40-transformed human fibroblasts by Sephadex G-10 gel filtration chromatography. The cells were first depleted of folates and then provided with limiting amounts of [3H]-folic acid in order that the cells would accumulate only forms of folate necessary for proliferation. Both the normal and the transformed cells accumulated monoglutamate and polyglutamate forms, but by 72 hours of labeling the transformed cells contained 3-10 times more polyglutamate than the normal cells. The growth rates for the normal and transformed cells were similar at this limiting folic acid concentration. Thus, if folate polyglutamates are more important for the proliferation of SV40-transformed cells than the normal cells, then inhibition of polyglutamate formation may be an important potential target for chemotherapy. PMID- 6274883 TI - Effect of H+ on the kinetics of Na+-dependent amino acid transport in Ehrlich ascites tumor cells: evidence for H+ as an alternative substrate. AB - The effects of H+ on the kinetics of alpha-aminoisobutyric acid (AIB) influx in Ehrlich ascites tumor cells have been investigated at different external Na+ concentrations. Elevation of [H+] in the presence of both high (154 mEq/l) and low (10 mEq/l) external Na+ leads to decreases in the maximum influx (Jmax) and increases in the apparent Michaelis-Menten constant (Km) for influx of AIB. In the virtual absence of external Na+ (0.96 +/- 0.04 mEq/l), alterations in [H+] are without measurable effect on AIB flux. Furthermore, addition of AIB (10 mM) to cell suspensions (pH 5.90) stimulates H+ uptake by the cells in either the presence or absence of Na+. The data are consistent with two kinetic models for Na+-dependent amino acid transport: an order bireactant (Na+-binding necessary before AIB binding) system or a random bireactant system. Both models require that H+ serve as an alternative substrate for Na+. The consistency of the models was tested by fit to data from the present study (not used to evaluate the kinetic parameters) and by prediction of the pH dependence of Na+-dependent amino acid transport compared to earlier studies. PMID- 6274884 TI - Differentiation of Dictyostelium discoideum mutant cells in a shaken suspension culture and the effect of cyclic AMP. AB - In Dictyostelium discoideum, 16 mutants in which cells differentiate into spores and stalk cells without normal morphogenesis were isolated. All these mutants are rapidly developing and capable of differentiating in a shaken suspension of phosphate buffer. The developmental kinetics of specific activities of enzymes in one of the mutants, HTY 1851, cultured in the suspension was compared with that in the parental strain, X2, developed on a solid surface. Most of the enzyme activities appeared much earlier and the peaks of the activities were lower in HTY1851 than X2, but the order or appearance of the activities was the same in both the strains cultured under the conditions described above. These results suggest that the biochemical steps in the development of the mutant in a shaken suspension are essentially the same as those of the parental strain X2 on a solid surface. It was also found that addition of cyclic AMP (2.5 X 10-5 M to 1 X 10-4 M) to the mutant cell suspension 6-8 h after the initiation of development induced an increase in the number of spores and the specific activities of some enzymes to values twice as high as those of an untreated control. PMID- 6274885 TI - Phagocytosis by macrophages. II. The dissociation of the attachment and ingestion steps. AB - The phagocytic process of mouse peritoneal macrophages was dissociated, using bovine serum albumin (BSA)-coated particles containing spin-labelled cholestanone, into 2 steps: attachment of particles to the cell surface and ingestion of the particles into the cytoplasm. The number of particles was estimated from electron spin resonance (e.s.r.) measurements. The particles ingested into the cytoplasm were distinguished from those attached to the cell surface by treatment with a membrane-impermeable reducing agent, ascorbate. The validity of the assay method was tested under various conditions. The measurements provided accurate and reproducible data. The phagocytic reaction was followed as a function of time and the rate constants for the attachment and ingestion steps were obtained from the initial phase. Both steps were highly dependent on temperature. Divalent cations in the incubation medium were essential for the attachment step but apparently had no effect on the ingestion step. The metabolic inhibitors, KCN and 2-deoxyglucose, inhibited both steps. Cytochalasin B inhibited both steps, while colchicine inhibited only the attachment step but apparently had no effect on the ingestion step. PMID- 6274886 TI - [Fistula-type pararectal mucinous carcinoma. A case report (author's transl)]. AB - A case of the exceptionally rare fistula type of pararectal mucinous carcinoma is reported, the patient presenting with clinical signs of a chronic abscess in the ischiorectal hollow. The unusual gelatinous secretion suggested the need for a biopsy of the fistula cavity. Pathological examination confirmed the diagnosis, where as a barium enema and rectoscopy gave negative results, eliminating the possibility of a digestive tract origin for the tumour. Local recurrence occurred two and a half years after an abdominoperineal resection, and was treated by local excision. Aetiopathogenic hypotheses and current therapeutic procedures are discussed. PMID- 6274887 TI - Isolation and characterization of three new polymyxins in polymyxins B and E by high-performance liquid chromatography. AB - Two polymyxin antibiotics, polymyxins B and E (colistin), have been separated analytically into ten to thirteen components on a commercial reversed-phase material by isocratic elution with a mixture of acetonitrile, phosphate/formate and acetate buffer containing sodium sulphate and triethylamine. The analytical system was transferred to a preparative system, using a C18-bonded stationary phase, without extensive impairing of the selectivity. The major components of each product were isolated and characterized by high-performance liquid chromatography, amino acid analysis and identification of the fatty acid. Three components were isolated and characterized for the first time. The fatty acid was also identified in some of the minor components. PMID- 6274888 TI - Use of periodate and methylamine for the quantitation of intracellular 5-fluoro 2'-deoxyuridine-5'-monophosphate by high-performance liquid chromatography. AB - High-performance liquid chromatography was used for the quantitation of [3H]fluorouracil and metabolites in L1210 cells with or without pre-treatment with methotrexate. Ribonucleotide pools were evaluated on a chemically bonded anion-exchange column. Deoxynucleotide pools were determined following periodate and methylamine treatment which eliminated the ribonucleotides (greater than 99.9% complete) and allowed for a rapid quantitation of the acid-soluble 5-fluoro 2'-deoxyuridine-5'-monophosphate (FdUMP) pools. FdUMP levels as low as 10 fmol/10(6) cells were detectable by polystyrene anion-exchange chromatography. Confirmation of these FdUMP values was performed by rechromatography on a reversed-phase ion-pair column. The use of periodate and methylamine as a means for the reliable elimination of the ribonucleotides will allow accurate isolation of deoxynucleotides by high-performance liquid chromatography. PMID- 6274889 TI - Silica Sep-Pak preparative chromatography for vitamin D and its metabolites. PMID- 6274890 TI - Determination of cefotaxime and desacetylcefotaxime in plasma and urine by high performance liquid chromatography. AB - A high-performance liquid chromatographic method is described for the analysis of the anti-bacterial agent cefotaxime and desacetylcefotaxime in physiological fluids. Plasma or serum samples were mixed with chloroform--acetone to remove proteins and most lipid material. The aqueous phase was then freeze-dried, reconstituted in mobile phase and chromatographed on a reversed-phase column using UV detection at 262 nm. Urine was analysed directly after centrifugation to remove particulate matter. The detection limit was 0.5--1.0 micrograms/ml for serum and 5 micrograms/ml for urine. The method has been applied to the analyses of cefotaxime and desacetylcefotaxime in plasma, serum, urine, cerebrospinal fluid, saliva, and pus from infected wound secretions. Two additional metabolites, which are lactones in which the beta-lactam ring has been opened, could be separated by this method. PMID- 6274891 TI - Assay of 5-aminosalicylate and its acetylated metabolite in biological fluids by high-performance liquid chromatography on dynamically modified silica. PMID- 6274892 TI - Acute effects of propylthiouracil (PTU) on thyroidal iodide organification and peripheral iodothyronine deiodination: correlation with serum PTU levels measured by radioimmunoassay. PMID- 6274893 TI - A new in vitro assay for human thyroid stimulator using cultured thyroid cells: effect of sodium chloride on adenosine 3',5'-monophosphate increase. AB - A new sensitive in vitro assay for human thyroid stimulator (HTS) was developed using human thyroid adenoma cells in monolayer culture. After being cultured for 2 days, the cells were incubated in 0.3 ml Hank's solution without 0.8% NaCl (medium 1) and with thyroid stimulator (bovine TSH or 3 mg patient serum immunoglobulin G) at 37 C for 2 h. The cAMP generated in the cells and the medium during the incubation was measured by RIA. The assay was sensitive enough to elicit a 1.7- to 7.9-fold increase in cAMP at a TSH concentration of 10 microU/ml. HTS was detected in 33 (82.5%) of the 40 patients with untreated graves' disease using this assay system. In Hank's solution (medium 2), however, HTS was detected in only 5 (23.8%) of the 21 patients with untreated GRaves' disease. cAMP increment upon stimulation by either TSH or HTS in medium 1 was larger than that in medium 2, and the difference in the response to HTS using the two media was much greater than that in the response to TSH. Therefore, all HTS immunoglobulin G studies showed higher activity using medium 1 than using medium 2 when expressed as bovine TSH equivalent. Analysis by the Lineweaver-Burk plot of dose-response curves of the effect of TSH and HTS stimulation on cAMP increment showed an increase in the Km upon the addition of NaCl to the medium. A similar inhibitory effect of NaCl (150 mM) was also observed in the assay system of human thyroid adenylate cyclase stimulator using crude plasma membrane fractions. In summary: 1) an assay for HTS measuring cAMP production in cultured thyroid adenoma cells was developed and the assay using low NaCL medium was found to be the most sensitive, and 2) the inhibitory effect of NaCl on the response to HTS was much greater than that on the response to TSH. These data suggest different behaviors of these two stimulators at their receptor sites. PMID- 6274894 TI - Sodium salicylate augments the plasma adrenocorticotropin and cortisol responses to insulin hypoglycemia in man. AB - To test the hypothesis that prostaglandins attenuate neuroendocrine responses to changes in circulating glucose levels in man, we studied the effects of sodium salicylate (SS), a prostaglandin synthesis inhibitor, on the plasma ACTH and cortisol responses to insulin hypoglycemia. Six normal men were given insulin (0.05 U/kg, iv) on 2 different days during the infusion of either SS (40 mg/min) or saline. Compared to the saline control, SS had no significant effect on either the rate of fall of plasma glucose after insulin or the glucose nadir (mean +/- SEM, 33 +/- 3 vs. 36 +/- 3 mg/dl; P = NS). Peak ACTH levels after insulin were higher during SS compared to those during saline in all six subjects (316 +/- 95 vs. 102 +/- 26 pg/ml; P less than 0.05), and SS had a clear effect to increase both the overall ACTH response (F = 21.3; P less than 0.01, by analysis of variance) and the plasma cortisol response (F = 6.72; P less than 0.05, by analysis of variance). The most striking example of this effect of SS occurred in one subject whose peak plasma ACTH was only 44 pg/ml during saline but reached 750 pg/ml during SS despite an identical fall of plasma glucose to 42 mg/dl. Augmentation of the ACTH and cortisol responses to insulin hypoglycemia may be the result of an alteration by SS of recognition of glucose levels by glucose sensitive cells of the brain, and effect which could be due to the inhibition of prostaglandin synthesis. PMID- 6274895 TI - Multiple forms of immunoreactive beta-endorphin are present in an ectopic adrenocorticotropin-producing tumor but not in normal pituitary or pituitary adenomas. AB - Human ACTH-producing tumor and plasma have been examined by gel filtration and ion exchange chromatography to detect the possible presence of reported multiple forms of immunoreactive beta-endorphin (I-EP) Ion exchange chromatography of I-EP obtained from gel filtration showed four components of I-EP [two major peaks in the positions of EP-(1-31) and EP-(1-27) and two minor peaks in the positions of N-acetyl EP-(1-31) and N-acetyl EP-(1-27)] in two ectopic ACTH-producing lung cancers, and two components of I-EP [the major peak in the position of EP-(1-31) and minor peak in the position of N-acetyl EP-(1-31) in an ectopic ACTH-producing thyroid cancer. Only a single peak in the position of EP-(1-31) was present in plasma from a patient with Nelson's sindrome and a patient with Addison's disease, in the pituitary adenomas from six patients with Cushing's disease, and in the nontumorous pituitary tissues from a patient with Cushing's disease and a patient with acromegaly. These data suggest that the posttranslational processing of EP in human pituitary is different from that in the ectopic ACTH-producing tumor. PMID- 6274896 TI - Isolated adrenocorticotropin deficiency associated with polyglandular failure. AB - A female patient presented with isolated ACTH deficiency and diabetes mellitus. During the course of a year's replacement with hydrocortisone, goiter developed, which, on biopsy, was diagnosed as typical Hashimoto's thyroiditis. Four years later, she experienced tetany, and the diagnosis of idiopathic hypoparathyroidism was made. Immunological studies revealed the presence of antithyroid antibodies in the serum and cell-mediated immunity to the thyroid and parathyroid. These data strongly suggest that the autoimmune process is involved in the genesis of some, though not all, forms of isolated ACTH deficiency. PMID- 6274897 TI - Adrenocortical function in old age: response to acute adrenocorticotropin stimulation. AB - To elucidate the controversial point whether, in analogy with delta 5-steroid secretion, adrenal delta 4-steroid secretion is significantly decreased in elderly persons, we studied the response of plasma levels of both delta 5 steroids (dehydroepiandrosterone, 17-hydroxypregnenolone, and pregnenolone) and delta 4-steroids (cortisol, androstenedione, 17-hydroxyprogesterone, and progesterone) to acute ACTH stimulation in four groups of young and elderly males and females, respectively. To study the possible influence of sex hormones on adrenocortical function in elderly persons, we performed the same study in male with prostatic carcinoma treated with estrogens. The data show that in elderly persons, the response of plasma delta 4-steroids to ACTH stimulation is comparable or higher than that in young subjects, whereas the response of delta 5 steroids is significantly decreased; estrogen treatment does not change this pattern. It is concluded that in elderly persons adrenal delta 4-steroid secretory capacity is unimpaired, whereas delta 5-steroid secretion is significantly decreased. The responsible mechanism remains to be elucidated. PMID- 6274898 TI - Human chorionic gonadotropin subunit measurement in primary hyperparathyroidism. AB - The concentration of subunits of hCG (hCG alpha and hCG beta) was determined in plasma or serum from 70 patients with primary hyperparathyroidism (1 degrees HPT). Two of three patients with parathyroid carcinoma showed elevation in plasma concentrations of both subunits, which fell after surgical removal of the tumor. An extract prepared from the tumor of 1 of these patients contained the subunits in high concentrations, whereas in extracts similarly prepared from tissues removed from 7 patients with benign 1 degrees HPT, the subunits were not detectable or were present in much lower concentrations. In 42 cases of benign 1 degrees HPT, samples from veins containing a 10-fold gradient of parathyroid hormone obtained during selective venous catheterization and peripheral samples from the same patients were analyzed for hCG subunits. Only 1 patient demonstrated a mild elevation of hCG alpha in parathyroid venous effluent alone that may have represented subunit release by apparently benign parathyroid tissue. Thirty patients with multiple endocrine neoplasia type I were tested; 8 evidenced clinically active islet cell tumors, and 6 of these 8 showed high circulating concentrations of hCG alpha (and hCG beta in 1 case). Patients in the multiple endocrine neoplasia type I group with 1 degrees HPT or pituitary tumors, but no evident pancreatic islet disease, did not show elevations in subunit concentrations. Thus, in patients with 1 degrees HPT, determination of subunits of hCG may be helpful in making the diagnosis of parathyroid carcinoma or in screening for associated (and probably malignant) pancreatic islet disease. PMID- 6274899 TI - Adrenocorticotropin-secreting pituitary adenomas originate from the anterior or the intermediate lobe in Cushing's disease: differences in the regulation of hormone secretion. PMID- 6274900 TI - Binding of steroids to mineralocorticoid receptors: implications for in vivo occupancy by glucocorticoids. PMID- 6274901 TI - Nature of thyrotropin displacement activity in subacute thyroiditis. AB - Using a radioreceptor assay and serum immunoglobulin (Ig) prepared by ammonium sulfate precipitation, significant TSH displacement activity (TDA) was demonstrated in 5 of 15 patients with subacute thyroiditis tested during the acute phase. Using a cAMP generation assay, adenyl cyclase stimulation by Ig from patients with subacute thyroiditis was not demonstrated. The nature of the TDA demonstrated in subacute thyroiditis was investigated to determine whether the factor measured was TSH receptor antibody, as is found in Graves' hyperthyroidism, or thyroglobulin, which is know to give false positive responses in the radioreceptor assay. When Ig was prepared by DEAE+-Sephadex chromatography, mean TSH displacement indices were similar to those given by ammonium sulfate-prepared Ig for both Graves' disease and subacute thyroiditis. On the other hand, when Ig was prepared by DEAE+-cellulose chromatography, which isolates highly purified IgG, mean indices were significantly less than for ammonium sulfate-prepared Ig for both Graves' hyperthyroidism and subacute thyroiditis. Thyroglobulin was not detected in Ig prepared by any of the 3 methods. Although high concentrations of crude thyroid-soluble fraction and purified thyroglobulin gave strongly positive responses in the radioreceptor assay, concentrations of thyroglobulin over the range found in the sera of patients with subacute thyroiditis could not be shown to give positive responses. Moreover, TSH displacement indices did not correlate with serum thyroglobulin levels. As determined by species cross-reactivity and dose-responses studies, the TDAs demonstrated in subacute thyroiditis and Graves' hyperthyroidism were similar. It was concluded that the TDA demonstrated in subacute thyroiditis represents antibody which binds to, but does not stimulate, the TSH receptor. PMID- 6274902 TI - Presence of immunoreactive gamma-melanocyte-stimulating hormone, adrenocorticotropin, and beta-endorphin in human gastric antral mucosa. AB - gamma MSH-like, ACTH-like, and beta-endorphin-like immunoreactivities (gamma MSH LI, ACTH-LI, and beta-endorphin-LI, respectively) were detected in water extracts of four human gastric antral mucosa. The concentrations of gamma MSH-LI, ACTH-LI, and beta-endorphin-LI in the boiling water extracts were 9.9 +/- 3.3, 6.2 +/- 1.8, and 3.9 +/- 1.3 ng/g (mean +/- SE), respectively. Gel exclusion chromatography on a Bio-Gel P-60 column showed that most ACTH-LI and beta endorphin-LI were eluted at the elution positions of human ACTH and beta endorphin, respectively. The major peak of gamma MSH-LI was eluted at the elution position of big gamma MSH-LI, but another peak was eluted at the elution position of small gamma MSH-LI, as in bovine intermediate pituitary. Concanavalin A agarose affinity chromatography showed that 52% of gamma MSH-LI was specifically bound to the column, but only 8% of ACTH-LI and none of beta-endorphin-LI were specifically bound. These results suggest that there exists an ACTH/beta lipotropin common precursor protein in human antral mucosa and that the processing of the precursor is accelerated as a bovine intermediate pituitary, indicating possible roles of these peptides in the function of the gastrointestinal tract. PMID- 6274903 TI - A case of pituitary adenoma with possible simultaneous secretion of thyrotropin and follicle-stimulating hormone. AB - A TSH- and FSH-screening pituitary adenoma was demonstrated in a 23-yr-old man who presented with bitemporal hemianopsia, but without clinical or laboratory evidences of hyper- or hypothyroidism or hypogonadism. Basal TSH (29-45 microunits/ml) and FSH (44-63 mIU/ml) were moderately elevated. GH and ACTH secretion were impaired, and basal PRL and testosterone were normal. TRH and LRH elicited a significant increase in TSH and FSH, respectively. The plasma glycoprotein alpha-subunit concentration was normal, and its response to simultaneous administration of TRH and LRH was low. Plasma TSH was not suppressed completely by exogenous T3. After operation and radiotherapy, elevated TSH and FSH as well as failure of T3 to suppress TSH were corrected. No significant changes in thyroid hormone concentration in the blood were detected in spite of the reduction of TSH. TSH and FSH concentrations in the tumor extract were lower than those in normal pituitaries. Histologically, the tumor was very vascular and consisted mostly of a single type of cell. The tumor cells were positively stained for FSH beta-subunit by immunohistochemical study, but for none of other pituitary hormones or subunits examined. Electron microscopic examination revealed relatively abundant single type secretory granules of high electron density. It is possible that the tumor simultaneously secreted TSH and FSH, the former possibly being immunologically active, but biologically inactive. PMID- 6274904 TI - Cushing's disease: transient secondary adrenal insufficiency after selective removal of pituitary microadenomas; evidence for a pituitary origin. PMID- 6274905 TI - Steroid and adenosine 3',5'-monophosphate formation in granulosa and thecal cells from human preovulatory follicles in response to human chorionic gonadotropin. AB - Granulosa and thecal cells from the preovulatory follicles of 14 women were mechanically isolated and separately incubated for short term periods (0.5-4 h) in the presence and absence of hCG. After incubation, tissue cAMP levels and medium progesterone content of (P), androstenedione (A), and 17 beta-estradiol (E2) were determined. All follicles appeared healthy and mature, as judged by their number of granulosa cells, histological examination of the oocytes, athe steroid levels in antral fluid, as well as the appearance of the oocyte-cumulus complexes. Under basal conditions, both granulosa and thecal cells had the capacity to synthesize all of the steroids measured. The predominant steroid formed by the granulosa cells was P, while the thecal cells formed A as the major steroid. Both cell types produced considerable amounts of E2. The addition of hCG in various concentrations gave rise to concentration-dependent increase in cAMP formation in both cell types. Furthermore, hCG caused a statistically significant increase in the formation of P and A by the thecal cells, while steroid formation by the granulosa cells was not significantly altered. With higher concentrations of hCG, however, there was a tendency toward a stimulation of P synthesis in the granulosa cells from most of the follicles tested. It is concluded that in preovulatory follicles of human origin, follicular steroidogenesis is not rigidly compartmentalized between the two cell types. Furthermore, both granulosa and thecal cells are sensitive to stimulation with hCG in this type of follicle. PMID- 6274906 TI - Testosterone secretion by cultured arrhenoblastoma cells: suppression by a luteinizing hormone-releasing hormone agonist. AB - A patient with virilization was studied. The basal urinary excretion of 17 ketosteroids was at the upper limit of normal, but the plasma testosterone concentration was greatly elevated. Testosterone secretion could be stimulated by hCG, suppressed by dexamethasone, and was not affected by ACTH. At operation, an arrhenoblastoma of the left ovary was found. Isolated tumor cells in culture secreted testosterone. The addition of a LRH agonist (10 ng/ml) suppressed the secretion of testosterone by 50% (P less than 0.01). The inhibiting effect of a LRH agonist on steroidogenesis suggests that LRH receptors were present on this tumor and that treatment with LRH agonists might be beneficial in patients with metastatic steroid hormone-secreting ovarian and testicular tumors. PMID- 6274907 TI - Acute dopaminergic inhibition of aldosterone secretion is independent of angiotensin II and adrenocorticotropin. AB - To clarify whether dopaminergic inhibition of aldosterone secretion is physiologically dependent on stimuli from tropic hormones, we attempted to block angiotensin II (AII)- and ACTH-mediated increases in the plasma aldosterone concentration (PAC) with dopamine in normal human subjects. The effect of dopamine on metoclopramide-induced aldosterone secretion also was studied. Six normal male subjects in metabolic balance on a 150 meq/day sodium and 60 meq/day potassium intake received AII infusion at 2, 4, and 6 pmol/kg . min, each dose for 30 min, on each of 2 consecutive days. On the first day, the subjects received a vehicle infusion from 60 min before to the end of the AII infusion; on the second day, dopamine (4 micrograms/kg . min) was substituted for vehicle. AII in the presence of vehicle increased PAC from 5.4 +/- 1.3 to 19.9 +/- 2.9 ng/100 ml; AII in the presence of dopamine increased PAC from 4.8 +/- 0.5 to 19.5 +/- 1.8 ng/100 ml (P = NS). After an interval of 3 weeks on an ad libitum diet, the same protocol was repeated except that ACTH (5, 10, and 20 U/h) was substituted for AII. ACTH in the presence of vehicle increased PAC from 8.1 +/- 2.7 to 27.3 +/- 3.1 ng/100 ml; in the presence of dopamine, ACTH increased PAC from 4.7 +/- 0.5 to 34.9 +/- 6.1 ng/100 ml (P = NS). Metoclopramide increased PAC from 4.5 +/- 0.6 to 17.8 +/- 2.3 ng/100 ml in the presence of vehicle and from 4.4 +/- 0.5 to 7.2 +/- 0.7 ng/100 ml in the presence of dopamine (P less than 0.01). Dopamine did not decrease basal PAC. Dopamine inhibits increases in aldosterone secretion induced by dopamine antagonist but does not alter AII- or ACTH-induced steroid secretion. Acutely, dopaminergic inhibition of aldosterone secretion is independent of AII and ACTH. PMID- 6274908 TI - Neurophysin biosynthesis in vitro in oat cell carcinoma of the lung with ectopic vasopressin production. AB - The incorporation of labeled compounds into neurophysins of a transplantable human oat cell carcinoma of the lung with ectopic vasopressin production was studied in vitro. Neurophysins in cell extracts and in incubation media were isolated by immunoprecipitation and analyzed by sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis. When cells were incubated with L [35S]cysteine for 12 h, SDS-polyacrylamide gel electrophoresis of the immunoprecipitates from cell extract and medium resolved two forms of neurophysins with apparent molecular mass of 10,000 (10K) and 20,000 (20K). Both forms of [35S]-neurophysins were completely displaced from the immunoprecipitates by excess human neurophysin. Incubation of cells with L-[35S]cysteine and D-[3H] glucosamine hydrochloride revealed that glucosamine was incorporated into the 20K neurophysin region, but not into 10K species. To observe the kinetics of labeling of the two forms of neurophysins, cells were incubated with L[35S]cysteine for varying periods of time. After short labeling periods, most of the radioactivity resided in 20K species, which plateaued after 1 h, whereas 10K neurophysin progressively increased in its height. When cells were chased with unlabeled cysteine after the exposure to a short pulse of labeling, 20K neurophysin peak gradually decreased with an apparent initial half-life of 1 h. In contrast, the label in 10K neurophysin steadily increased, which exceeded the former by 3 h of chase. Analysis of 20K neurophysin in cell extract by isoelectric focusing on polyacrylamide gel demonstrated that it was principally composed of a protein with an apparent isoelectric point (pI) of 5.7. These results suggest that neurophysin is synthesized in ectopic vasopressin-producing tumors by post translational processing from a glycosylated proneurophysin with an apparent molecular mass of 20,000 daltons and a pI of 5.7. PMID- 6274909 TI - Hypothyroidism modulates beta adrenergic receptor adenylate cyclase interactions in rat reticulocytes. AB - We have investigated alterations in beta adrenergic receptor binding sites of rat reticulocytes occurring in animals rendered hypothyroid by thyroidectomy. Beta adrenergic receptor interactions were assessed by measuring the number of (-)[3H] dihydroalprenolol binding sites and the ability of an agonist to compete for occupancy of the receptors. The number of receptors was significantly reduced in cells from the hypothyroid animals. In addition, there were significant agonist specific alterations in binding. Using computer assisted curve fitting techniques, it was found that the ability of (-)isoproterenol to stabilize a high affinity guanine nucleotide sensitive "coupled" form of the receptor was impaired. Reticulocytes from hypothyroid animals have, in addition, a reduction in the concentration of the nucleotide regulatory protein as assessed by the number of 42,000 Mr substrates for cholera toxin catalyzed ADP ribosylation. These alterations are associated with reductions in catecholamine and NaF stimulated adenylate cyclase activity. Diminished coupling of beta adrenergic receptors with other regulatory components of the adenylate cyclase system represents a mechanism by which altered thyroid states modulate beta adrenergic receptor function and beta adrenergic responsiveness of tissues. PMID- 6274910 TI - Glucose and triiodothyronine both induce malic enzyme in the rat hepatocyte culture: evidence that triiodothyronine multiplies a primary glucose-generated signal. AB - We have stimulated in a cultured hepatocyte system the synergistic interaction between triiodothyronine (T3) and dietary carbohydrate in the induction of malic enzyme (ME). Kinetic studies revealed that isolated hepatocytes equilibrate with media T3 within 5 min; nuclei equilibrate with media T3 by 45 min; and the half time of T3 metabolism was 10 h in 10% serum. We demonstrated nuclear T3 receptors in isolated hepatocytes and the induction of ME by T3 in physiological concentrations. However, in the complete absence of T3 glucose could still induce ME. At all concentrations of glucose (100-1,000 mg/dl), T3 (0.3 nM free T3) resulted in a relatively constant (1.4- to 1.7-fold) increase in ME response. The augmentation in ME activity was paralleled by an enhanced rate of enzyme synthesis as determined by [3H]leucine incorporation into immunoprecipitable ME. Cells cultured in serum free media also demonstrated a glucose-dependent increase in ME. Insulin greatly stimulated the glucose induction of ME, whereas dexamethasone had very little influence on ME induction. These studies demonstrate the usefulness of an adult hepatocyte tissue culture model for the study of the effects of T3 on gene expression in cells that are not derived from tumor. They clearly demonstrate that well established effects of T3 can be simulated in such a system at levels of free hormone that approximate those in extracellular body fluids. Our results indicate that an increased concentration of glucose per se can induce the formation of ME in the absence of alterations in extrahepatic hormones or factors. Moreover, our findings confirm inferences from in vivo studies that T3 acts as a multiplier of a glucose-induced signal. PMID- 6274911 TI - Secondary effect of aldosterone on Na-KATPase activity in the rabbit cortical collecting tubule. AB - The possibility that mineralocorticoids have a direct influence on renal Na-K ATPase activity has been the focus of intense research effort and some controversy for a number of years. Early studies were hindered by an inability to differentiate between possible glucocorticoid vs. mineralocorticoid effects on this enzyme within the multitude of cells that comprise the heterogeneous mammalian nephron. This study attempts to circumvent this problem by monitoring Na-K ATPase activity in the rabbit renal cortical collecting tubule (CCT), a proposed target epithelium for mineralocorticoids. Using an ultramicro assay, Na K ATPase activity was measured in CCT from normal, adrenalectomized (adx), and adx rabbits subjected to one of several corticosteroid treatment protocols. The results indicate that Na-K ATPase activity in the CCT decreased by 86% subsequent to adrenalectomy. Injection of physiological doses of aldosterone (10 micrograms/kg) but not dexamethasone (100 micrograms/kg) restored CCT Na-K ATPase activity in adx rabbits to normal levels within 3 h after injection. An insignificant rise in activity was observed 1.5h after aldosterone treatment. In addition, spirolactone SC 26304, a specific mineralocorticoid antagonist, blocked the action of aldosterone on Na-K ATPase.. Therefore an acute increase in Na-K ATPase activity participates in the action of aldosterone on Na transport in this segment. To differentiate between primary vs. secondary activation of this enzyme, adx animals were treated with amiloride before the injection of aldosterone with the intent of blocking luminal membrane Na entry into CCT. In these animals, pretreatment with amiloride blocked the increase in CCT Na-K ATPase act activity seen with aldosterone alone at 3 h. Thus the increase in activity with aldosterone appears to be a secondary adaptation that is dependent on an aldosterone-enhanced increase in the passive entry of Na across the luminal membrane. The subcellular mechanism by which Na modulates Na-K ATPase activity remains obscure. PMID- 6274912 TI - Apolipoprotein C-III-1 activates lysosomal sphingomyelinase in vitro. AB - Apolipoprotein (apo)C-III-1 from human very low density lipoprotein stimulates 14 fold the activity of lysosomal sphingomyelinase from human fibroblasts. At the sphingomyelin concentrations tested, maximal stimulation was obtained with 5 microM apoC-III-1 or apoC mixture. Apolipoproteins A-I, A-II, B, and C-I conferred little or no stimulation. Sphingomyelinase was stimulated 20-fold by lysophosphatidylcholine with an optimum concentration of 70 microM using 0.3 mM substrate. Sphingomyelinase activity was inhibited by concentrations of apoC-III 1 and lysophosphatidylcholine three- to fivefold above stimulatory levels. Triton X-100 activated sphingomyelinase 300-fold with a pH optimum of 5.0, while the pH optimum with the biological activators was 4.0. These results raise the possibility of an in vivo activity for the biological activators. The proteins that enter lysosomes as part of a lipoprotein complex may activate lysosomal enzymes that degrade the lipid components. PMID- 6274913 TI - Hydrogen ion secretion by the collecting duct as a determinant of the urine to blood PCO2 gradient in alkaline urine. AB - Several theories have been advanced to explain the elevation in urinary PCO2 during bicarbonate loading and include: (a) H+ secretion, (b) countercurrent system for CO2, (c) the "ampholyte" properties of bicarbonate, and (d) mixing of urine of disparate bicarbonate and butter concentrations. In this study microelectrodes were used to measure in situ and equilibrium pH (pHis and pHeq) and PCO2 in control and bicarbonate loaded rats before and after infusion of carbonic anhydrase. The disequilibrium pH method (pHdq = pHis - pHeq) was used to demonstrate H+ secretion. Control rats excreting an acid urine (pH = 6.04 +/- 0.06) failed to display a significant disequilibrium pH at the base (BCD), or tip (TCD) of the papillary collecting duct. Urine pH (7.54 +/- 0.12), and urine to blood (U-B) PCO2 increased significantly during NaHCO3 loading while PCO2 at the BCD and TCD also increased (95 +/- 4 and 122 +/- 4). Furthermore, an acid disequilibrium pH was present at both the BCD and TCD (-0.42 +/- 0.04 and -0.36 +/- 0.03) and was obliterated by carbonic anhydrase. Comparison of the PCO2 in the BCD or TCD with the adjacent vasa recta revealed similar values (r = 0.97). It is concluded that H+ secretion by the collecting duct into bicarbonate containing fluid with delayed dehydration of H2CO3, is the most likely determinant of the U-B PCO2 in alkaline urine. Similar values for PCO2 in the collecting duct and the adjacent vasa recta suggests trapping of CO2 in the medullary countercurrent system. The rise in PCO2 occurs both along the collecting duct and after exit from the papilla. PMID- 6274914 TI - Isolated nephron segments from rabbit models of obstructive nephropathy. AB - Micropuncture and microcatheterization studies have been used extensively to investigate the pathophysiologic alterations in renal function induced by urinary tract obstruction. The present isolated tubule microperfusion studies were designed to examine the intrinsic alterations in segmental nephron function induced by 24 h of bilateral (BUO) and unilateral (UUO) urinary tract obstruction. Following UUO superficial proximal convoluted tubule reabsorption rate (J(v)) was not different from contralateral control (0.75+/-0.08 vs. 0.73+/ 0.11 nl/mm per min, NS). Following UUO J(v) in juxtamedullary proximal convoluted tubules (JMPCT) was reduced 32% (0.69+/-0.06 vs. 0.47+/-0.04 nl/mm per min, P < 0.02). Following UUO J(v) in proximal straight tubules (PST) was reduced 52% (0.25+/-0.02 vs. 0.12+/-0.03, P < 0.01). Thick ascending limb (T-ALH) function was assessed by measurement of ability to lower perfusate chloride ion concentration (DeltaCl). Following UUO DeltaCl was reduced 76% (-39+/-9 vs. -9+/ 1 meq/liter, P < 0.001). Cortical collecting tubule (CCT) function was assessed by measurement of antiduiretic hormone (ADH)-dependent osmotic water flow. Following UUO osmotic water flow was reduced 76% (0.90+/-0.08 vs. 0.22+/-0.04 nl/mm per min, P < 0.01) and this ADH resistance could not be overcome by cAMP. Nephron segments were then examined following relief of BUO. There were no differences in intrinsic function following relief of BUO when compared with UUO. We conclude that in UUO and BUO (a) the intrinsic tubular defects are identical, (b) the natriuresis noted is due, in part, to disordered JMPCT, PST, and T-ALH NaCl reabsorption, (c) the impaired concentrating ability is due, in part, to depressed function in T-ALH and ADH resistance of the CCT, and (d) the ADH resistance occurs at a site distal to the intracellular generation of cAMP. PMID- 6274915 TI - Decreased adrenal responsiveness to angiotensin II: a defect present in spontaneously hypertensive rats. A possible model of human essential hypertension. AB - 30% of patients with essential hypertension have a decreased adrenal response to angiotensin II (A II) on a low but not a high sodium intake. They also have a compensatory increase in the activity of the renin-angiotensin system best documented in a sodium-restricted state.To assess whether such a mechanism could account for the hypertension in genetically hypertensive rats, adrenal responsiveness to A II was determined in three groups of rats; spontaneously hypertensive rats (SHR), normotensive Wistar rats (WKY), and normotensive Sprague Dawley rats (SDR). Animals in each group were placed on either a low or high sodium diet for 14 d with balance assessed by sodium excretion. The animals were then decapitated, blood was obtained for plasma renin activity (PRA), A II and aldosterone and adrenals isolated for the preparation of purified glomerulosa cells. The cells were incubated in Krebs-Ringer bicarbonate solution, containing bovine serum albumin, for 60 min in the absence and presence of increasing concentrations of A II. The PRA, basal aldosterone output, and adrenal sensitivity to A II were similar in the three groups of rats on the high sodium diet. On the low sodium diet the SHR had a significantly (P < 0.01) higher PRA (25+/-7 ng/ml per h) than either the WKY (12+/-2 ng/ml per h) or the SDR (7+/-1 ng/ml per h) and lower basal aldosterone output (68+/-17 vs. 154+/-43 and 197+/ 21 ng/10(6) cells per h, respectively). In addition, the slope of the A II dose response curve was more shallow (P < 0.01) in the cells from the SHR than those obtained from the WKY and SDR.Thus, the SHR PRA and aldosterone responses to sodium restriction and aldosterone response to A II were similar to that previously described in a subgroup of patients with essential hypertension suggesting that the SHR will serve as a model for exploring the mechanism(s) responsible for the hypertension in these patients. PMID- 6274916 TI - Functionally abnormal Na+-K+ pump in erythrocytes of a morbidly obese patient. AB - The Na(+)-K(+) pump in the erythrocytes of a mordibly obese patient shows a unique constellation of functional abnormalities. The number of pump units, measured by [(3)H]ouabain binding to intact cells, as well as the enzymatic activity of the (Na(+)-K(+))-dependent ATPase in erythrocyte membranes were found to be markedly increased compared with control cells (18-fold and 14-fold, respectively). There was a concomitant fivefold increase in the rate of pump mediated uptake of (86)Rubidium (a K analogue); this was balanced by an increased rate of (86)Rb efflux. In striking contrast to normal cells, however, a major portion of this efflux (80%) was inhibited by ouabain, and thus appeared to be mediated by the Na(+)-K(+) pump. Erythrocytes from this patient had elevated levels of intracellular K(+) and reduced levels of intracellular Na(+). This finding, taken together with the ouabain inhibition of K(+) efflux and the absence of associated abnormalities, argues against the possibility that the increased number of Na(+)-K(+) pump units was a compensation for a primary increase in the permeability of the erythrocyte membrane to monovalent cations, as is seen in a variety of erythrocyte disorders. Further evidence for a primary abnormality of the enzyme was our observation that the cardiac glycoside ouabain bound to these cells with reduced affinity and had a right shifted dose response for pump inhibition. The markedly increased number of Na(+)-K(+) pump units in these cells did not appear to extend to mononuclear leukocytes.In conclusion, the erythrocytes from this patient have a very large number of functionally abnormal Na(+)-K(+) ATPase units. A unique abnormality of the erythrocyte Na(+)-K(+) ATPase of these cells is the most likely explanation for these findings. PMID- 6274917 TI - Modern diuretics and the kidney. PMID- 6274918 TI - Identification and localization of the motor nuclei and sensory projections of the glossopharyngeal, vagus, and hypoglossal nerves of the cockatoo (Cacatua roseicapilla), Cacatuidae. AB - Horseradish peroxidase (HRP) has been applied to the proximal severed ends of glossopharyngeal (N IX), vagus (NX), and hypoglossal (N XII) cockatoo in order to localize the motoneurons and sensory projections of these nerves which are involved in the control of the bird's feeding and phonatory behaviors. Application of HRP to N IX labeled four rhombencephalic nuclei: (1) a large celled, retrofacial nucleus supplying M. geniohyoideus, the major tongue extensor; (2) a dorsal nucleus composed of medium-sized cells, projecting to most branches of N IX; (3) a ventrolateral nucleus supplying, amongst other structures, the floor of the pharynx and larynx; and (4) a ventral portion of the dorsal motor nucleus of the vagus. Neurons labeled by application of HRP to the cervical vagus comprise the classically defined dorsal motor nucleus and a ventrolateral medullary nucleus which is coextensive with that of the glossopharyngeus: together they probably constitute a nucleus ambiguus. Application of HRP to hypoglossal branches labeled a large nucleus intermedius (IM) and neurons ventral, ventrolateral, and caudal to it. The rostral third of IM supplies the lingual muscles, the caudal two-thirds the tracheosyringeal muscles. Many labeled neurons were found in the "jugular" ganglion following HRP treatment of each of the three nerves, especially N IX and N XII, which innervate the tongue. Central projections of these neurons are to nuclei of the descending trigeminus and to largely nonoverlapping portions of the principal trigeminal nucleus. It is hypothesized that these afferents provide sensory information necessary for the efficient processing and passage of food in the mouth. PMID- 6274919 TI - Efferent projections of the medial preoptic nucleus and medial hypothalamus in the pigeon. AB - The efferent projections of the medial preoptic nucleus (POM), anterior-medial hypothalamic area (AM), and the posteromedial hypothalamic nucleus (PMH) in the pigeon were traced by the autoradiographic technique. Similar and differential connections were noted from these regions. Projections from POM and AM-PMH were traced to nucleus septalis lateralis, nucleus dorsomedialis thalami, nucleus dorsolateralis anterior thalami (pars ventralis), posterior hypothalamic and medial mammillary areas, area ventralis tegmenti (Tsai), central gray of midbrain and nucleus intercollicularis and substantia grisea periventricularis of the midbrain. The density of silver grains in these regions differed with POM and AM PMH injections. Other projections were observed exclusively from only one or two of the nuclear regions injected. Connections from POM and the rostral part of AM were seen to the median eminence, neurohypophysis, and the nucleus of anterior pallial commissure. Only cells of the anterior part of AM project fibers to nucleus septalis medialis. In the hypothalamus, projections from POM are concentrated in the periventricular region and in the preoptic-hypophyseal tract in the extreme lateral hypothalamus, while AM-PMH projections are heaviest in the medial hypothalamus and lateral preoptic area. A major difference in the connections of PMH from POM is the more substantial PMH projection to the midbrain. A prominent projection courses dorsolaterally and posteriorly from PMH toward nucleus ovoidalis and splits into two pathways: a lateral pathway which heavily innervates n. intercollicularis and the periventricular gray and a ventrolateral projection to the midbrain tegmentum. The projections described above provide anatomical substrates for neuroendocrine, autonomic, and behavioral functions. PMID- 6274920 TI - Retinal projections in the goldfish: a study using cobaltous-lysine. AB - Cobaltous lysine applied to the distal stump of a severed optic nerve was used to study the retinal projections of normal adult goldfish. Both the termination areas of optic axons and the pathways they traveled were established. Contrary to previous descriptions of the goldfish visual system, the optic nerves do not decussate completely at the optic chiasm. Fascicles that entered the ipsilateral optic tract innervated targets in the ipsilateral thalamus and optic tectum. Other optic fibers crossed the posterior commissure from the contralateral side of the brain and also innervated the ipsilateral tectum and thalamus. In addition, optic fibers bilaterally innervated a hypothalamic target in close proximity to the infundibulum that may correspond to the nucleus tuberis lateralis. The contralateral preoptic region contained two discrete areas of innervation, each served by separate fascicles. The ipsilateral preoptic region was similarly innervated, but more sparsely. Fibers that entered the controlateral ventral thalamus originated from three fascicles and terminated in three distinct targets. In contrast, three targets in the contralateral dorsal thalamus were served by one fascicle, and fibers passed from one nucleus to the other two. Innervation of the ipsilateral thalamus was similar to that seen contralaterally. Each main optic tract divided into three tracts, two of which entered the optic tectum, while the other innervated several pretectal areas. Other fibers innervated an accessory optic nucleus located near nucleus glomerulosus. The contralateral tectum contained numerous radially oriented optic fascicles. These fascicles represented optic fibers that left thalamic and pretectal targets to enter the optic tectum from beneath the stratum periventriculare. Optic fibers were also observed in the transverse commissure, tractus rotundus, horizontal commissure, tectobulbar tract, and fasciculus retroflexus. Therefore, it appears that many of the anomalous projections seen after tectal ablation or after optic nerve crush are not in fact aberrant. Such projections probably reflect the presence of unusually large numbers of optic fibers in tracts that normally contain optic axons, as well as increased innervation of areas that normally receive sparse retinal projections. Filled tectal cells that could represent cells projecting to the retina were not observed in either tectal lobe. The ipsilateral retinal projections could not be attributed to cobaltous-lysine being transneuronally transported in readily detectable amounts. PMID- 6274921 TI - The afferent and efferent organization of the lateral geniculo-prestriate pathways in the macaque monkey. AB - Both anterograde (autoradiographic) and retrograde (horseradish peroxidase) tracing techniques were used to identify and characterize projections from the dorsal lateral geniculate nucleus (DLG) of the thalamus to the subdivisions of occipital prestriate cortex (generally defined by areas 18 and 19) in macaque monkeys. It was found that the DLG projects to area 19 and the anterior portion of area 18 located on the lateral, ventral, and ventromedial surfaces of the hemisphere. There is a general topographical organization such that the medial portion of the DLG projects to dorsal prestriate located between the lunate and superior temporal sulci, while the lateral portion of the DLG projects to ventral prestriate extending from the inferior occipital to the occipitotemporal sulci. The DLG projects to most of the occipitotemporal sulcus. In contrast DLG inputs to the lunate, superior temporal, and inferior occipital sulci are limited in extent, and involve only a portion of the bank or shoulder of each sulcus which is continuous with the cortex located on the surfaces of the surrounding preoccipital gyri. The projection to the inferior occipital sulcus is more extensive than the ones to the lunate and superior temporal sulci and involves the floor as well as the ventral bank. This means that there are certain functional subdivisions, such as those located within the lunate (eg, visual area 2) and superior temporal sulci which do not receive DLG input. Regardless of the location of these projections the terminal pattern was the same and occurred in horizontally segregated "patches" which were restricted to layer V and the lower, adjacent portion of layer IV. In contrast, the projections from the nearby pulvinar to cortical layers IV, III, and I of the same prestriate areas do not overlap with those from the DLG. An attempt was made to identify possible afferent inputs to these DLG-prestriate paths. The HRP experiments reveal that the DLG-prestriate cells are concentrated in the DLG interlaminar zones and that their distribution overlaps the distribution of terminals from the superior colliculus and, as shown previously, prestriate cortex. Intraocular injections of radioactive precursors demonstrated transsynaptic transport to a number of structures except prestriate cortex. While the latter result does not prove that the DLG-prestriate cells do not receive retinal input, one conclusion is that the DLG-prestriate projection could be organized like pulvino-prestriate systems which receive their inputs from the midbrain and cortex. PMID- 6274922 TI - The organization of centrifugal projections from the anterior olfactory nucleus, ventral hippocampal rudiment, and piriform cortex to the main olfactory bulb in the hamster: an autoradiographic study. AB - The centrifugal projections from the various subdivisions of the anterior olfactory nucleus (AON) can be categorized into four groups based on the organization of terminal fields in the main olfactory bulb (MOB). Pars lateralis and dorsalis have bilaterally asymmetric laminar projections to the MOB. The ipsilateral projections terminate primarily in the superficial half of the granule cell layer and in the deep third of the glomerular layer, whereas the contralateral projections terminate primarily in the superficial half of the granule cell layer and do not extend into the glomerular layer. Pars ventralis and posterior have bilaterally symmetric laminar projections with heavy terminations both in the superficial half of the granule cell layer and in the deep third of the glomerular layer. Pars medialis sends predominantly ipsilateral projections to the deep half of the granule cell layer. Pars externa has predominantly contralateral projections with a very narrow terminal field immediately deep to the internal plexiform layer. The projections to the MOB from the ventral hippocampal rudiment (HR) and the piriform cortex (PC) are exclusively ipsilateral. The projections from the ventral HR terminate primarily in the deep half of the granule cell layer. The projections from the PC also terminate predominantly in the granule cell layer, but there is a progressive shifting of terminal fields from the superficial half of this layer toward deeper regions for centrifugal axons arising from progressively more caudal levels of the PC. The laminar termination patterns of cortical afferents to the ipsilateral MOB thus are correlated with the mediolateral axis of the olfactory peduncle and the rostrocaudal axis of the piriform cortex. The centrifugal axons from these various sources enter directly into the granule cell layer of the caudal MOB or pass through the internal plexiform layer of the accessory olfactory bulb to reach the middle and anterior part of the MOB. We have termed these two routes the final common bulb pathway. The centrifugal axons from the laterally situated sources join the anterior and bulbar limbs of the anterior commissure before entering the final common bulbar pathway. In contrast, the centrifugal axons from pars medialis and the ventral HR travel diffusely in the cellular layer of the ipsilateral olfactory peduncle. A small component of the centrifugal projections from the PC travels in association with the lateral olfactory tract. PMID- 6274923 TI - Cholinergic and catecholaminergic afferents to the olfactory bulb in the hamster: a neuroanatomical, biochemical, and histochemical investigation. AB - A series of neuroanatomical, biochemical, and histochemical studies have been conducted to determine the sources of cholinergic afferents to the main olfactory bulb (MOB) in the hamster. Following horseradish peroxidase (HRP) injections that are restricted to the MOB, retrograde neuronal labeling is observed bilaterally in the anterior olfactory nucleus, locus coeruleus, and raphe nuclei, and ipsilaterally in the ventral hippocampal rudiment, dorsal peduncular cortex, piriform cortex, nucleus of the lateral olfactory tract, anterior pole of the medial septal area and vertical limb of the diagonal band, nucleus of the horizontal limb of the diagonal band (HDB), and hypothalamus. Spread of HRP into the accessory olfactory bulb results in additional neuronal labeling ipsilaterally in the bed nucleus of the accessory olfactory tract, medial amygdaloid nucleus, and bed nucleus of the stria terminalis, and bilaterally in the posteromedial cortical amygdaloid nucleus. Retrograde tracing studies also have been conducted in cases with lesions in the basal forebrain or hypothalamus to assess the extent to which such lesions interrupt fibers of passage from other sources of centrifugal afferents, and the effects of such lesions on choline acetyltransferase (CAT) activity and catecholamine content in the MOB and on acetylcholinesterase (AChE) activity in the forebrain have been evaluated. Lesions in the basal forebrain reduce or eliminate CAT and AChE activity in the MOB in direct relationship to the extent of damage to the HDB. Norepinephrine (NE) content in the MOB also is reduced by basal forebrain lesions, but in relationship to damage of the medial forebrain bundle (MFB). The hypothalamic lesions have no effect on AChE activity in the forebrain or on CAT activity in the MOB, but they eliminate retrograde labeling in the locus coeruleus and raphe nuclei and reduce the NE content of the MOB to undetectable levels. The dopamine content of the MOB is not reduced by any of the lesions. Anterograde tracing studies have been conducted to compare the rostral projection patterns of the HDB with the distribution of AChE activity. Most of the rostrally directed axons travel in association with the MFB. A small component of axons travels in association with the lateral olfactory tract. Within the MOB, the axons terminate predominantly in the glomerular layer and in the vicinity of the internal plexiform layer. The projection and termination patterns of the HDB correspond well with the distribution of AChE activity. These various results indicate that the HDB is the major source of cholinergic afferents to the MOB. PMID- 6274924 TI - Toxicity of diminazene aceturate (Berenil) to camels. PMID- 6274925 TI - Pathogenesis of pigeon herpesvirus infection. PMID- 6274926 TI - Herpesvirus as a cause of fatal disease in Australian wallabies. PMID- 6274927 TI - Vertical transmission of Border disease infection. PMID- 6274928 TI - Histochemical and ultrastructural features of an equine pulmonary granular cell tumour (myoblastoma). PMID- 6274929 TI - Malacoplakia-like lesion in the lymph node of a pig. PMID- 6274931 TI - Evaluation of blood-brain barrier permeability changes in rhesus monkeys and man using 82Rb and positron emission tomography. AB - Dynamic positron tomography of the brain with 82 Rb, obtained from a portable generator [82Sr (25 days) - 82 Rb (76 sec)], provides a means of studying blood brain barrier (BBB) permeability in physiological and clinical investigations. The BBB in rhesus monkeys was opened unilaterally be intracarotid infusion of 3 M urea. This osmotic barrier opening allowed entry into the brain of intravenously administered rubidium chloride. The BBB opening was demonstrated noninvasively using 82Rb and positron emission tomography and corroborated by the accumulation of 86Rb in tissue samples. Positron emission tomography studies can be repeated every 5 min and indicate that dynamic tomography or static imaging can be used to study BBB permeability changes induced by a wide variety of noxious stimuli. Brain tumors in human subjects are readily detected because of the usual BBB permeability disruption in and around the tumors. PMID- 6274930 TI - The pathogenesis of natural and simulated natural foot-and-mouth disease infection in cattle. PMID- 6274932 TI - CT differentiation of solitary from diffuse bronchioloalveolar carcinoma. AB - The therapy and prognosis of bronchioloalveolar carcinoma vary greatly with the solitary versus the diffuse form of the disease. Solitary disease demonstrates a high resectability rate with good long term prognosis. This contrasts with the rapidly fatal course associated with the diffuse form. To date, categorization of patients into either the solitary or the diffuse form has been based solely on conventional radiography. Multiple authors have reported cases of disease not demonstrated radiographically but discovered at surgery or autopsy. With the superiority of chest computed tomography (CT) for demonstrating parenchymal abnormalities, we propose that preoperative CT may be crucial in the workup of patients with presumed solitary bronchioloalveolar cell carcinoma. Early identification of diffuse disease or confirmation of the presence of solitary disease by CT may allow the institution of proper therapy and better evaluation of patient prognosis. PMID- 6274933 TI - Differential diagnosis of hepatic masses on computed tomography, with particular reference to hepatocellular carcinoma. AB - Differential diagnosis was attempted in 225 cases of hepatic masses using computed tomographic (CT) criteria. Hepatocellular carcinomas were suggested by the presence of an isodense mass, a narrow circular zone surrounding the mass, bulging of the tumor from the hepatic surface, decreased attenuation of an entire hepatic lobe, and diffuse homogeneous enhancement following bolus injection. The presence of liver cirrhosis on CT also favored the diagnosis of hepatocellular carcinoma. More than half the metastatic tumors showed findings rarely encountered in hepatocellular carcinomas, i.e., more than 10 lesions, masses with nodular margins and showing a gradual decrease in density toward the center, multiple calcifications, and peripheral enhancement following bolus injection. All cavernous hemangiomas were correctly diagnosed by serial scanning after bolus injections. Hepatic abscesses also revealed pathognomonic CT findings, while cholangiocarcinoma did not disclose characteristic features. Rapid serial scanning after the bolus injection of contrast medium was very helpful in the differential diagnosis of hepatic masses by CT. PMID- 6274934 TI - CT demonstration of fat tissue in malignant renal neoplasms: atypical Wilms' tumors. AB - It has been implied that the recognition of fat in a renal tumor suggests that the lesion is benign. Several authors have suggested tht angiomyolipoma is the only common renal tumor containing mature adipose tissue. Other neoplasms may contain mature fat that can be identified with current diagnostic imaging techniques. Under discussion is our experience with Wilms' tumor containing a predominance of adipose tissue. PMID- 6274935 TI - Effect of adrenocorticotropin on milk and plasma cortisol and prolactin concentrations. AB - Milk cortisol and prolactin concentrations were measured in 12 lactating Holstein cows for 3 days (Experiment 1). On day 3, cows were divided randomly into groups: IV1, IV4, IM, and control. Group IV1 received one intravenous injection of 49 IU adrenocorticotropin, group IV4 the same dosage four times at 2-h intervals, group IM 240 IU adrenocorticotropin intramuscularly, and control cows were injected with saline. Cortisol concentrations in blood plasma were increased in all groups following adrenocorticotropin injection, while milk cortisol concentrations increased fourfold in groups IV4 and IM and remained unchanged in group IV1. In Experiment 2, infusion of 18% saline into the mammary gland of three cows increased the somatic cell count in the infused quarter but had no effect on prolactin and cortisol of milk or plasma. In Experiment 3, six cows each were assigned to treatment groups of saline controls, IV2-0 (40 IU adrenocorticotropin intravenously at 0 and 2 h post-milking) or IV2-8 (40 IU adrenocorticotropin intravenously at 8 and 10 h post-milking). Half udders of each cow were milked before treatment and 4 h later. Increases of cortisol in plasma increased cortisol concentrations of milk 4 h after each treatment (IV2-0 and IV2-8). However, by 12 h after treatment in IV2-0 cortisol concentrations of milk had returned to normal. Increases in adrenal cortisol secretion are followed rapidly (within 4 h) by increased cortisol concentrations in milk. However, these decline rapidly in the absence of sustained increments in blood cortisol. Increased cortisol concentrations in milk most likely represent sustained elevation in plasma cortisol. PMID- 6274936 TI - Melanotic neuroectodermal tumour of the maxilla. PMID- 6274937 TI - A study of beta-adrenergic and prostaglandin receptors in patients with aspirin induced bronchospasm. AB - Nine patients with aspirin-induced bronchospasm were compared with age-matched normal controls to test the hypothesis that a defect in the beta-adrenergic system could lead to excessive dependence on prostaglandins to maintain bronchodilator function. Lymphocyte beta-receptor density and affinity were measured by 3H-dihydroalprenolol binding. Beta-receptor function was measured in terms of cyclic AMP accumulation in response to isoproterenol. Prostaglandin (PG) responsiveness was measured in terms of cyclic AMP accumulation caused by PGE1 stimulation. No difference was found in either ther number of beta-adrenergic receptors (1234 +115 vs 1213 +82) or dissociation constant (1.40 +0.23 nM vs 1.19 +0.07nM) (mean +SEM) between the asthmatic and the control group, respectively. Furthermore, neither group showed any difference in maximally stimulated cyclic AMP in response to isoproterenol and PGE 1. Thus we find no evidence of an inherent defect of beta-adrenergic receptor and function to account for the susceptibility to aspirin-induced bronchospasm. PMID- 6274938 TI - The limits of sponsorship for women physicians. PMID- 6274939 TI - The actress and the doctor. PMID- 6274940 TI - Attitudes of single parents toward health issues. PMID- 6274941 TI - The health and social service picture. PMID- 6274942 TI - Isoniazid toxicity as a cause of peripheral neuropathy. Case presentation and discussion. PMID- 6274943 TI - Neurilemoma (schwannoma) as a cause of tarsal tunnel syndrome. Case report and review. PMID- 6274944 TI - [Comparative study of tissue cultures from normal and pathological corneal epithelia (author's transl)]. AB - Changes in corneal epithelium in macular dystrophy are secondary. Those in lattice dystrophy are primary, the epithelial cells showing the characteristics of cancer cells as well as hypertrophy of the lysosomal system and vacuolisation with autolysis. After infection by herpes virus the epithelial cells show an increase in number and volume of the lysosomes, which liberate enzymes with cytolytic effects. PMID- 6274945 TI - Apert's syndrome: (acrocephalosyndactylism). AB - A case history is presented that illustrates the clinical and radiographic findings of an Apert's syndrome patient. The incidence of this condition at birth is approximately 1:160,000 with a greater frequency found in the children of older parents. A brief comparison is made with two similar syndromes. Carpenter's syndrome and LMBB syndrome. Recognition and diagnosis of each syndrome is difficult due to the phenotypic overlap and is achieved through an understanding of the similarities as well as the difference. In the Apert's syndrome patient, the hand and foot deformities present particular problems regarding everyday performance. The hands of these patients are generally surgically corrected to improve hand function through removal of the syndactylism or partial amputation. However, this was not performed on this patient for he was able to write legibly and had good pincer grasping ability. The feet in the Apert's syndrome patient present unique problems due to the malformations that exist rather than acquired deformities because of excessive joint motion. The structure of the foot is generally compatible with conventional shoewear. If the patient's psychological status warrants it, elective surgical intervention would be indicated to remove hypertrophic bone at weight-bearing areas to relieve pressure. PMID- 6274946 TI - The 24-h urinary cyclic adenosine 3', 5' monophosphate/creatinine ratio: an useful approach to the diagnosis of parathyroid disorders and function. AB - 24-h urinary cyclic adenosine 3', 5'-monophosphate/creatinine (cAMP/Cr) ratio was assessed in 10 patients with hypoparathyroidism, 6 with primary hyperparathyroidism, 7 with normocalcemic hypercalciuria and recurrent nephrolithiasis, 14 with osteomalacia, 25 with Paget's disease and 53 with symptomatic postmenopausal osteoporosis. In hypoparathyroid subjects the mean values of 24 h cAMP/Cr ratio were significantly lower than the control values, whereas in patients with parathyroid adenoma the mean values were higher and fell after parathyroid surgery. Patients with nephrolithiasis due to absorptive hypercalciuria showed low or normal cAMP/Cr ratio, whereas in those with osteomalacia and mean values of cAMP/Cr ratio were significantly higher than the control values and decreased after vitamin D treatment. The mean value of the 24 h urine cAMP/Cr ratio was normal in patients with Paget's disease or postmenopausal osteoporosis and increased significantly after long term treatment with calcitonin or diphosphonate. This increase paralleled a significant decrease of calcium plasma level. A significant improvement of fractional calcium absorption was observed in women with postmenopausal osteoporosis at the end of treatment with calcitonin or diphosphonate. PMID- 6274947 TI - Seasonal variation in urinary excretion of cyclic AMP in healthy people. AB - In a group of healthy young men and women the daily urinary excretion of cyclic adenosine 3', 5'-monophosphate (cAMP), calcium, hydroxyproline and the renal threshold phosphate concentration were evaluated at monthly intervals during 1 yr. A significant seasonal variation in cAMP urinary excretion was demonstrated, with a maximum occurring in spring and a minimum in winter. A clear annual rhythm was also observed when the other above parameters were considered. These findings are of importance in the interpretation of urinary cAMP values in clinical situations, and in the study of bone metabolism. PMID- 6274948 TI - Pharmacology of inhibitors of parietal cell function. PMID- 6274949 TI - Studies on camelpox virus. PMID- 6274950 TI - Relationship between peroxisomes and endoplasmic reticulum investigated by combined catalase and glucose-6-phosphatase cytochemistry. AB - The theoretical advantages of electron microscopic cytochemistry were utilized to look for evidence of possible connections between peroxisomes and the endoplasmic reticulum in rat liver. Established cytochemical procedures for catalase (peroxisomes) and glucose-6-phosphatase (endoplasmic reticulum) were carried out, and evidence was sought of diffusion of reaction products between the organelles. No such diffusion was observed: lead phosphate was found in the endoplasmic reticulum and in the nuclear envelope but not in peroxisomes; oxidized diaminobenzidine (DAB) was seen only in peroxisomes. In addition, both types of cytochemistry were carried out on the same tissue. The two kinds of reaction product could be distinguished by virtue of their different electron opacities. No mixing of the two reaction products was observed. These results do not support the hypothesis that peroxisomes and endoplasmic reticulum may be connected; rather, they support the idea that the two organelles exist as separate cellular compartments. PMID- 6274951 TI - Autoradiographic demonstration of uptake and retention of 3H-estradiol after in vitro incubation. AB - An in vitro incubation method is described for the demonstration of 3H-estradiol in sections of mouse uterus by thaw-mount autoradiography. The method involves the incubation of tissue sections in 5 nM 3H-estradiol with a subsequent 2 hr chase in medium containing 3.5 g% bovine serum albumin. The distribution of the silver grains observed compares favorably to that seen by others with dry-mount autoradiography after in vivo injection. The labeling is inhibited by excess estradiol or diethylstilbesterol, but not by progesterone or hydrocortisone. Its subcellular distribution appears predominantly nuclear in presumptive target cells. Some regional variability in degree of labeling is present throughout the sections, but is far less marked within a given area. Because the observed labeling has been retained during a 2 hr chase and can be inhibited, it is likely to represent physiologically significant uptake and retention of 3H-estradiol in target cells. Preliminary results with human mammary carcinoma suggest this method may be applicable to the investigation of estrogen target cells in human tissue. PMID- 6274952 TI - Localization of cyclic adenosine monophosphate phosphodiesterase in mouse alveolar cells. PMID- 6274953 TI - Histochemical and ultrastructural characterization of subendothelial glycoprotein microfibrils interacting with platelets. AB - The interaction of human blood platelets with collagenase-treated rabbit subendothelium was studied by histochemical ultrastructural methods and by morphometric semi-quantitative analysis. Aortas were deendothelialized and incubated: 1) with a highly purified bacterial collagenase whose specificity was controlled; and 2) with the same collagenase followed by chymotrypsin. For histochemical studies, tannic acid, ruthenium red, and peroxidase-labeled Ricinus communis and concanavalin A were used. Electron microscopy showed that after digestion of fibrillar collagen by collagenase, adherent and aggregated platelets were observed on Ricinus communis-, concanavalin A-, and ruthenium red-positive glycoprotein microfibrils. After successive incubation with collagenase and chymotrypsin, the microfibrils disappeared. No platelets were observed on the remnant amorphous elastin. Morphometric analysis confirmed the interaction of platelets with collagenase-treated subendothelium. In addition, glycoproteins were extracted from collagenase-treated rabbit aortas using 5 M guanidine. Using an in vitro quantitative test, significant platelet adhesion to these glycoproteins was observed. Our results show an interaction between platelets and noncollagenic glycoprotein microfibrils. PMID- 6274954 TI - Cultured human monocytes require exposure to bacterial products to maintain an optimal oxygen radical response. AB - Freshly isolated human blood monocytes displayed a vigorous oxygen radical response, measured as release of superoxide anion (O2-), after stimulation with phorbol myristate acetate (PMA) or opsonized zymosan. High O2- release was observed with cells isolated by using a variety of procedures. Monocytes cultured in endotoxin-free medium M199 with or without 5% heat-inactivated autologous serum gradually lost this ability to produce O2- in response to PMA over the course of 4 days. The decreased responsiveness to PMA was accompanied by decreased adherence and viability. The loss of function, adherence, and viability was prevented by supplementing the culture medium with either bacterial lipopolysaccharide (LPS) or muramyl dipeptide (MDP). The O2- response of monocytes cultured for several days without bacterial products could be partially restored by the addition of LPS on day 2 or 3 of culture. Partial restoration could be detected in monocytes after only 1 hr of exposure to LPS, although a maximal response required a 2-day exposure. The minimum effective concentration of MDP was 1 ng/ml; stereoisomers of MDP, which are inactive as adjuvants, had no effect at 1 micrograms/ml. The minimum effective concentration of LPS was 1 pg/ml, corresponding to fewer than 10 molecules of LPS per monocyte. These results suggest that exposure to LPS or other bacterial products, represented here by MDP, may be required to preserve the microbicidal potential of human monocyte-macrophages in vivo. PMID- 6274955 TI - Immunoglobulin expression by human B lymphocytes clonally transformed by Epstein Barr virus. AB - Epstein Barr virus (EBV) immortalizes human B lymphocytes, resulting in long-term lymphoblastoid cell lines. To date, such cell lines have typically been derived by suspension culture of mixed mononuclear cells, infected in vivo or in vitro with EBV. We have derived 208 lymphoblastoid lines by clonal transformation of peripheral blood lymphocytes from 3 neonates, 3 normal adults, and 1 patient with systemic lupus erythematosus (SLE), utilizing colony formation in agarose and then separate propagation of the colonies in liquid medium. A total of 180 lines have been analyzed for expression of cytoplasmic and/or secreted immunoglobulin isotypes. Such analysis reveals that 167 (92%) of the lines are monoclonal with respect to isotype expression. All clonally derived lines from neonates produce only IgM. Most clonal transformants from normal adults also produce IgM (81%), but clones producing IgG (16%) or IgA (3%) were also found. Clonal transformation of lymphocytes from the SLE patient revealed a shift in isotype expression, with increased numbers of IgG-secreting clones (48%) and IgA-secreting clones (13%). The data support the use of EBV-mediated clonal transformation as a probe for the analysis of states of isotype differentiation among circulating B cells, and as a method for deriving immortalized, monoclonal human B cell lines. PMID- 6274956 TI - Association of Sendai virion envelope and a mouse surface membrane polypeptide on newly infected cells: lack of association with H-2K/D or alteration of viral immunogenicity. PMID- 6274957 TI - Pharmacologic modulation of release of arachidonic acid from human mononuclear cells and lymphocytes by mitogenic lectins. PMID- 6274958 TI - Establishment of human T lymphocyte clones highly cytotoxic for an EBV transformed B cell line in serum-free medium: isolation of clones that differ in phenotype and specificity. PMID- 6274959 TI - T cell-mediated immunoregulation of Epstein Barr virus- (EBV) induced B lymphocyte activation in EBV-seropositive and EBV-seronegative individuals. AB - Infection with Epstein Barr virus (EBV) is accompanied by seroconversion and life long persistence of the virus in B lymphocytes. During acute EBV-induced infectious mononucleosis, suppressor T cells become activated, which may provide an additional mechanism of host defense against the causative agent. When cultures of lymphocytes from normal adults seropositive for EBV were stimulated with the B95-8 strain of EBV, purified B cells produced increasingly higher numbers of immunoglobulin- (Ig) secreting cells, whereas in co-cultures of autologous B and T cells a profound suppressor T cell activity inhibited further B cell activation after 10 to 12 days in culture. No such T cell-mediated inhibitory effect was seen in cultures of lymphocytes obtained from normal adults seronegative for EBV, indicating a correlation between the suppressor effect with evidence of prior immunity to this virus. The T cell-mediated suppression in patients with infectious mononucleosis is characterized by an early-acting inhibitory effect on B cell differentiation that is not specific in that all polyclonal B cell activators are inhibited, whereas in EBV-seropositive normal subjects suppression is delayed in time and affects only EBV-activated cultures. These data indicate that after infection with EBV, immunoregulatory T cells are generated that are capable of inhibiting further EBV-induced activation of autologous B cells and thus may provide an additional unique mechanism of host defense against persisting EBV-infected B cells. PMID- 6274960 TI - Characterization of the secretory activity of leukotriene B4 toward rabbit neutrophils. AB - We have described in det ail the secretory activity of leukotriene B4 toward rabbit neutrophils. Leukotriene B4 rapidly and vigorously degranulates rabbit neutrophils. This activity is stereospecific, cytochalasin B-dependent, and is enhanced by extracellular calcium. Pretreatment with leukotriene B4 deactivates rabbit neutrophils, i.e., cells so treated do not respond to stimulation by an additional bolus of leukotriene B4. In addition, the secretory activity of leukotriene B4 is sharply dependent on the simultaneous presence of cytochalasin B. Rabbit neutrophils therefore exhibit the previously described desensitization to the effect of cytochalasin B. In these and other discussed respects the characteristics of the leukotriene B4-induced degranulation of rabbit neutrophils are strikingly similar to those of the chemotactic factors. These results support the hypothesis that leukotriene B4 mediates, at least in part, the secretory, and possibly other, activities of chemotactic factors. PMID- 6274961 TI - B lymphoblast antigen (BB-1) expressed on Epstein-Barr virus-activated B cell blasts, B lymphoblastoid cell lines, and Burkitt's lymphomas. AB - Two new cell surface antigens expressed on B lymphoblastoid cell lines (B-LCL) were defined with cytotoxic mouse monoclonal antibodies. One marker, BB-1 (for B lymphoblast antigen-1), was detected on human and nonhuman primate B-LCL, Epstein Barr virus (EBV)-activated B cell blasts, most Burkitt's lymphomas, and Ia+ B lymphoblast-like myelomas. Polyclonal B cell activators such as pokeweed mitogen (PWM) and lipopolysaccharide (LPS) also induced the expression of BB-1 on immunoglobulin (Ig)-positive cells. In contrast, BB-1 could not be detected on normal lymphoid tissues by complement-dependent cytotoxicity and immunofluorescence (IF) assays or by analysis with a fluorescence-activated cell sorter (FACS). T cell blasts, T cell leukemias, and pre-B cell or erythroblastic leukemia cell lines were also BB-1 negative. Of particular interest was the finding that BB-1 was expressed on the Jijoye lymphoma but only marginally on a subline of Jijoye, P3HR-1, that lacks receptors for EBV and produces a defective virus incapable of transforming lymphocytes. A second lymphoblast antigen (LB-1) unlike BB-1, was present on both T and B cell blasts and virus-transformed T- and B-LCL but not on normal lymphoid tissues. PMID- 6274962 TI - Abnormal anti-Epstein Barr virus antibodies in carriers of the X-linked lymphoproliferative syndrome and in females at risk. AB - The asymptomatic hemizygous female carriers of the X-linked lymphoproliferative syndrome (XLP) have abnormal antibody responses to EBV. This suggests partial expression of the defect that leads to EBV-provoked life-threatening diseases in their affected sons. EBV specific antibodies were measured in 65 serum samples of 12 obligate carrier females and seven of their daughters (females at risk) during periods ranging from 1 to 5 yr. Abnormal qualitative antiviral capsid antigen (VCA) IgG titers were nearly fourfold higher than normal controls, two carriers had persistent IgM anti-VCA antibody, two-thirds had persistent IgA anti-VCA antibody, and half of the women had titers to early antigen (EA). Five of seven females exhibited a similar persistent pattern. In contrast, none of the unaffected family members nor 23 normal controls expressed IgA or IgM titers to VCA even with high exposure to the virus, and anti-EA was detected in only one control. Therefore, these findings may prove useful for detecting carriers of the syndrome. Abnormal anti-EBV titers similar to the carrier pattern have been reported in patients and other immunosuppressed individuals, and are indicative of active viral infection. PMID- 6274963 TI - Correlation of human neutrophil secretion, chemoattractant receptor mobilization, and enhanced functional capacity. AB - Studies were performed to elucidate further the phenomenon of secretagogue mediated enhancement in the binding of the chemoattractant f-met-leu-[3H]phe to human neutrophils (PMN). Specific f-met-leu-[3H]phe binding to unstimulated PMN reached maximum levels after 10 to 15 min of incubation at 0 degrees C with a saturating concentration of peptide, and consisted of a readily displaceable and a nondisplaceable component. PMN, preexposed to A23187 (2.5 X 10(-8) M) or PMA (0.5 ng/ml) for 30 min at 37 degrees C to stimulate limited and preferential release of specific (secondary) granules (10 to 20% of total lysozyme, no beta glucuronidase), demonstrated an approximate doubling in the displaceable component of f-met-leu-["3H]phe binding, accompanied by an increasing nondisplaceable component that could not be explained by bulk pinocytosis of extracellular fluid (assessed by [3H]sucrose uptake). The increase in f-met-leu [3H]phe binding was not affected by inhibitors of protein synthesis, could not be attributed to the secreted products lysosyme or lactoferrin acting on the cell, and, on the basis of studies with PMN from patients with chronic granulomatous disease, could not be attributed to the effects of reactive oxygen species generated in low concentration during stimulation. Functional studies on PMN indicated that preexposure to secretagogues at concentrations demonstrated to increase receptor availability also enhanced subsequent f-met-leu-phe-mediated superoxide and hydrogen peroxide generation. The present data demonstrate that secretagogues may activate PMN to enhance their subsequent responses in f-met-leu phe-mediated processes, and, combined with previous reports, support the concept that specific granules provide a source of preformed membrane and receptor material that is translocated to the cell surface during the secretion associated with directed locomotion. PMID- 6274964 TI - The effect of ischemia on cyclic adenosine 3',5'monophosphate accumulation in mouse skin. AB - The incubation of adult mouse skin pieces in a buffered salts medium at 37 degrees C led to a rapid accumulation of cyclic adenosine 3'5-monophosphate (cyclic AMP) in the tissue. In mouse skin maximum accumulation occurred after 2 min incubation; levels reverted to near control levels after a further 7 min incubation. the increase in cyclic AMP contents of the skin pieces was probably not due to the release of materials which activate adenylate cyclase after binding to cellular receptors. Thus, cyclic AMP accumulation was unaffected by the inclusion of alpha- or beta-adrenergic antagonists, or by the pretreatment of adult mouse skin with indomethacin (an inhibitor of prostaglandin synthetase). Furthermore, adenosine, a known activator of epidermal adenylate cyclase, could not be detected in the incubation medium. The functional integrity of epidermal adenylate cyclase was maintained during the cyclic AMP accumulation in response to ischemia. Thus, adenosine, histamine, isoproterenol and prostaglandin E2 (PGE2) augmented the cyclic AMP response. Cyclic AMP accumulation at 37 degrees C was not observed in newborn mouse skin; this lack of cyclic AMP accumulation was probably not due to increased activity of low affinity cyclic AMP phosphodiesterase in newborn mouse skin. PMID- 6274965 TI - Resistance to cefamandole: a collaborative study of emerging clinical problems. AB - Cefamandole resistance in five patients was studied. Microorganisms emerged resistant to cefamandole during therapy with the drug in three patients with complicated infections. This resistance was associated with an enhanced production of beta-lactamase and/or with a change in the substrates and the isoelectric focusing patterns of the enzymes. Cross-resistance to other beta lactam antibiotics developed concurrently in isolates from these patients. Disk diffusion tests did not detect resistance to cefamandole in the pretreatment isolate from the fourth patient; this isolate produced inactivating enzymes, and resistance was detected only in broth dilution tests. In the fifth patient, infection with a cefamandole-resistant Enterobacter developed during postoperative therapy with the drug. Resistance to cefamandole in the isolate from this patient was unstable and was associated with inducible beta-lactamase activity. These examples emphasize the need for close monitoring of patients who are given cefamandole and for thorough in vitro evaluation of isolates from the patients both before and after treatment. PMID- 6274966 TI - Increased risk of infection with hepatitis A and B viruses in men with a history of syphilis: relation to sexual contacts. AB - Serum samples from 269 men with at least one episode of syphilis were tested for the presence of markers of hepatitis A and B viruses. The patients were divided into three groups: 177 homosexual men, 35 heterosexual men, and 57 men with unknown sexual contacts. The incidence of hepatitis B surface antigen or its antibody (anti-HBs) was three times greater in the homosexual men (67%) than in the men with unknown sexual contacts (25%) and the heterosexual men (23%). The incidence of anti-HBs increased with age, reaching 84% in the homosexual men 40 49 years of age. A positive correlation was found between the presence of markers of hepatitis A and B viruses and the number of episodes of syphilis in younger homosexual men (P less than 0.01). Antibody to hepatitis A virus was found in 36% of the homosexual men, 18% of the men with unknown sexual contacts, and 20% of the heterosexual men. Only one patient was found to have IgM antibodies to hepatitis A virus. PMID- 6274967 TI - Pleomorphic, enveloped, virus-like particles associated with gastrointestinal illness in neonates. AB - Pleomorphic, enveloped, virus-like particles were detected by electron microscopy in the stools of symptomatic infants during an outbreak of gastrointestinal illness in a neonatal intensive-care unit. To determine the incidence of virus like particles in the stool and their relation to gastrointestinal symptoms, eight surveys of stools for the particles were conducted over 40 weeks. The incidence of virus-like particles in the stool decreased from 69% to less than 10% over the study period. Most infants surveyed were premature; overall, 32 (36%) of 88 neonates were positive for virus-like particles. Statistically significant associations were found between virus-like particles in the stool and gastrointestinal symptoms within one week of each survey. These symptoms included water-loss stools, blood in the stool, gastric retention, bilious gastric aspirates, and abdominal distention. Several infants with virus-like particles whose mothers had gastrointestinal or "flu-like" symptoms before delivery were identified in the community (not part of the survey study). PMID- 6274968 TI - Persistent infection with adenovirus types 5 and 6 in lymphoid cells from humans and woolly monkeys. AB - Chronic infection with adenovirus types 5 and 6 was established in primary mononuclear leukocytes from human umbilical cord blood and in Epstein-Barr virus (EBV)-transformed B lymphocytes from human umbilical cord blood and from woolly monkey blood. Adenovirus could be recovered from cultures of primary leukocytes and of EBV-transformed lymphocytes for two and three months, respectively, without visible alteration of cell growth. Infection in cultures of EBV transformed lymphocytes from woolly monkey blood was obliterated by exposure to antibody, but EBV-transformed lymphocytes from human umbilical cord blood contained small amounts of virus for prolonged periods that restored infection in the culture when antibody was removed. Thus, chronic infection of lymphoid cells by some adenoviruses is maintained by at least two mechanisms: cell-to-cell spread of virus in the absence of antibody and intracellular persistence of infectious virus in the presence of antibody. PMID- 6274969 TI - Virus-specific IgG and IgM antibodies in normal and immunocompromised subjects infected with cytomegalovirus. AB - Levels of IgG and IgM antibodies t human cytomegalovirus (CMV) were measured using a solid-phase radioimmunoassay. Individuals positive by complement-fixation test consistently had detectable IgG titers by radioimmunoassay, but no quantitative relationship was apparent. An elevated IgM titer was considered specific for CMV infection because sera from individuals with other herpesvirus infections did not cross-react. In patients with mononucleosis, elevated titers of IgM antibody to CMV correlated (P less than 0.001) with active infection and were highest during viremia. Titers of IgG antibody to CMV during and after symptomatic infection were similar to those of asymptomatic positive individuals. Increases in CMV-specific IGM were observed in both primary and reactivated infections in cardiac transplant recipients. In a small group of cardiac transplant recipients with recurrent symptomatic disease, IgM titers were low at the time of viruria and did not increase with CMV tissue involvement, a result which suggests that quantitative deficiencies in IgM may be related to the severity of CMV infections. PMID- 6274970 TI - In vitro susceptibility of antimony-resistant Leishmania to alternative drugs. PMID- 6274971 TI - [Virucidal activity of alcohols. Virucidal efficiency of alcohols against viruses in liquid phase (author's transl)]. PMID- 6274972 TI - [Virucidal activity of alcohols. Virucidal efficiency of ethanol on viruses in organic materials (author's transl)]. PMID- 6274973 TI - [Primary Epstein-Barr virus infection in an aged patient (author's transl)]. PMID- 6274974 TI - [Infectious complications of bone marrow transplantation (author's transl)]. PMID- 6274975 TI - [Poly(ADP-ribose) polymerase and DNA (author's transl)]. PMID- 6274976 TI - [Studies on the prostaglandin E2 receptor in human corpora lutea (author's transl)]. AB - It is known that Prostaglandins (PGs) have a specific effect on the corpus luteum function. In this paper we have studied the physical and chemical characteristics of PGE2 receptor in human corpora lutea to explain the mechanism on corpus luteum function. The presence of PGs binding sites in corpora lutea has been reported, but hardly any papers are available on the presence of PGE2 binding sites in human corpora lutea. The present study was undertaken to determine, using the proportion graph method, whether PGE2 receptors are present in human corpora lutea or not. The number of binding sites and association constant of PGE2 receptor to human corpora lutea of the luteal phase of the cycle and gestation were examined. There is evidence (1) that PGE2 receptors were present in human corpora lutea, and the binding sites have one specific and one non-specific binding system. (2) That the number of binding sites in early, mid and late luteal phase corpora lutea were respectively 105-157, 190-196, 162-190 f moles/mg protein. Association constants were 2.0-12.0 x 10(11) M-1. (3) That the number of binding sites of 6th week gestation corpora lutea were 98-100 f moles/mg protein, and those of 12th week gestation were 40-85 f moles/mg protein. Association constants of the gestation corpora lutea were 17.0-20.0 x 10(11) M-1. PMID- 6274977 TI - [Cell functions of cultured human choriocarcinoma following the administration of methotrexate (author's transl)]. AB - Treatment of choriocarcinoma should be eradicated of trophoblastic cells which undergo destruction in both of primary and metastatic sites and Methotrexate (MTX) has a most effective value. While MTX is being administration to the patients, there is no determination of clear relationship between proliferation of trophoblastic cells and production of human chorionic gonadotropin (hCG). In this report, we attempted to clarify the changes of the dividing compartments in cell cycle of human choriocarcinoma in vitro in the administration of MTX (concentration in 5 x 10(-6) M and 5 x 10(-7) M) to the experimental cells. Cell samples were stained by propidium iodide (PI) and fluorescein isothiocyanate (FITC), then were measured nucleic acid and protein content by means of flow microfluorometry (FMF). MTX was remarkably increased the intensity of FITC associated with protein production, in dose that is not only killing effect to the cells but inhibition of DNA synthesis. Moreover, the most difference in both subjects were as follows; Ever when MTX in low concentration was removed after 48 hours, 50% of experimental cells recovered to the level of controls in contrast to the persistence of remarkable inhibition of DNA synthesis in high concentration. PMID- 6274979 TI - [A case of mixed mesodermal tumor of the ovary (author's transl)]. PMID- 6274978 TI - Development of ovarian tumors experimentally studied in mice by 60Co-ray irradiation. AB - Female C3H strain mice aged 5 to 6 weeks were exposed to a single dose of 400 Rad irradiation to their lower abdomens and the morphological changes of their ovaries were observed until 72 weeks after irradiation. The incidence of tumor development increased gradually from 34 weeks after irradiation till 72 weeks, when ovarian tumors were noted in more than 90% of the animals. Three types of ovarian tumors developed after a prolonged period, postirradiation: tubular adenoma, granulosa cell tumor and luteinizing tumor. Mixed tumor types account for 68% of the tissue examined. The tubular adenoma is believed to arise from the germinal epithelial inclusion cyst, the granulosa cell tumor and luteinizing cell tumor arise from the interstitial cells. In the case of the granulosa cell tumor, it is believed that tumor cells one capable of secreting estrogen, because vaginal smears demonstrated estrus and histological sections of endometrium demonstrated hypertrophy. PMID- 6274980 TI - Functional alpha 1 protease inhibitor produced by a human hepatoma cell line. AB - Alpha 1 protease inhibitor antigen was identified in the culture medium of the human ascites hepatoma cell line SK-HEP-1. Trypsin inhibitory activity and alpha 1 Pl antigen accumulated in serum-free medium concomitantly over a period of several days. Radioactive alpha 1 Pl antigen was detected in conditioned medium from cultures supplemented with 35S-L-methionine, indicating a synthesis and release of the protein. Alpha 1 Pl antigen in conditioned medium appeared to be antigenically identical to that in human plasma, and the newly synthesized (radiolabeled) antigen co-migrated with plasma, alpha 1 Pl after immunoelectrophoresis or SDS-polyacrylamide gel electrophoresis. Moreover, evidence is presented that the synthesized inhibitor exhibits functional activity, since the 35S-labeled alpha 1 Pl in conditioned medium complexes with trypsin. We conclude that SK-HEP-1 cells in culture produce functionally active alpha 1 Pl which may be identical to that in plasma. PMID- 6274981 TI - Cytotoxicity of the glycolipid region of streptococcal lipoteichoic acid for cultures of human heart cells. AB - The ability of LTA of Streptococcus pyogenes to stimulate cell division or to kill tissue culture cells derived from human heart was investigated. Initial studies indicated that at low concentrations, ranging from 0.01 to 1.0 micrograms/ml, LTA stimulated cell division, whereas at higher concentrations, ranging from 10 to 1000 micrograms/ml, it killed the cells. Deacylated lipoteichoic acid, which lacked cell membrane binding activity, similarly stimulated or killed the heart cells depending on the concentration added to the tissue cultures. Fractionation of LTA after mild ammonia hydrolysis yielded a polyglycerophosphate and polar lipid fraction, both of which retained the glucose from the glycolipid moiety of the LTA molecule, and a neutral lipid fraction that was devoid of phosphorus or glucose. Toxic activity was present only in the fractions containing glucose. Oxidation of LTA or its deacylated derivative with sodium metaperiodate reduced the assayable glucose content without destroying the polyglycerophosphate backbone and resulted in a parallel loss of cytotoxicity, strongly suggesting that the glucose moieties of S. pyogenes LTA must be intact for the toxic activity against human heart cells to be expressed. PMID- 6274982 TI - Lactoferrin interacts with deoxyribonucleic acid: a preferential reactivity with double-stranded DNA and dissociation of DNA-anti-DNA complexes. AB - LF was found to bind to deoxyribonucleic acid as assessed by immunofluorescence studies on cell nuclei, affinity chromatography of DNA on immobilized LF, and gel chromatography of an LF-DNA reaction mixture. LF immobilized on Sepharose 4-B was reacted with 125I-labeled DNA in both its double-stranded and single-stranded configurations; dsDNA eluted with a 0.69M NaCl buffer, whereas ssDNA eluted with a 0.25M NaCl buffer. Additional evidence for a preferential reactivity with dsDNA was provided by the enzymatic treatment of preformed dsDNA-LF and ssDNA-LF complexes with S1 endonuclease, and DNAse 1--DNase digestion alone liberated free LF. The interaction of LF with DNA partially inhibited the binding of anti-DNA antibodies from patients with SLE, as assayed in a standard Farr assay. Furthermore, DNA-anti-DNA (labeled with 125I-IgG) complexes could be dispersed in vitro by the addition of LF. It is hypothesized that the release of LF by neutrophils chemotactically attracted to DNA-anti-DNA complexes may act as a feedback loop to modulate the inflammatory response in SLE. PMID- 6274983 TI - Effect of potassium depletion on the vasopressin-sensitive cyclic AMP system in rat outer medullary tubules. AB - The effects of PDN on VP-sensitive cAMP metabolism were examined in MCT and MAL microdissected from the rat kidney. VP-sensitive adenylate cyclase activity was significantly reduced (delta -46%; p less than 0.05) in MAL of PDN rats but, in sharp contrast, was significantly increased (delta +79%; p less than 0.02) in MCT of PDN rats compared to controls. cAMP phosphodiesterase activity was significantly increased in both MAL (delta +59%; p less than 0.005) and MCT (delta +79%; p less than 0.001) of PDN rats compared to controls. The increase in cAMP accumulation in MAL measured in response to VP in intact tubules did not differ between PDN and controls, whereas cAMP accumulation in response to VP was significantly higher (delta +127%; p less than 0.001) in MCT of PDN rats compared to controls. The present results would indicate that the observed in vivo resistance to the antidiuretic effect of VP that occurs in PDN is not due to an impairment in VP-sensitive cAMP accumulation in MCT, but would rather suggest that a defect exists at a cellular step subsequent to cAMP generation. In addition, our results illustrate that the extent and directionality of in situ accumulation of cAMP measured in intact tubules cannot always be predicted from rhe activities of enzymes controlling its synthesis and degradation (adenylate cyclase and cAMP phosphodiesterase), which are measured in vitro in disrupted tubules. PMID- 6274984 TI - Contrasting patterns of protein kinase activities among normal peripheral blood lymphocyte subpopulations and lymphoid cell lines. AB - Human peripheral mononuclear cell suspensions were separated into E-rosette positive (T cell) and E-rosette-negative (non-T cell) fractions and assayed for cAMP binding activity and histone and casein kinase activity, in the presence and absence of cAMP. Each of these activities was higher in E-rosette-positive than E rosette-negative fractions. Cyclic AMP binding and histone kinase activities were further noted to be higher in T lymphoblastoid cell lines when compared to lines of non-T lymphoid cell origin. In addition. DEAE-cellulose chromatography revealed a qualitative contrast of isoenzymic composition of protein kinase from these cell lines. Cyclic AMP-independent casein kinase activity was high in both types of cell lines. These results suggest that high protein kinase activity is characteristic of T cells and may be related to their higher sensitivity to cAMP agonists in comparison to non-T cells. This study also provides further evidence for an association between increased cAMP-independent protein kinase activity and the transformed state, which has been noted for a variety of other cell types. PMID- 6274985 TI - Isolation of human cytomegalovirus from peripheral blood T cells of renal transplant patients. AB - Previous reports have demonstrated that human CMV can be isolated from the mononuclear leukocyte fraction of peripheral blood taken from patients excreting CMV. Many renal transplant patients show evidence of an active CMV infection, and in these patients T cell responses are often impaired. Thus the purpose of the current study was to determine whether the virus could be isolated from either the T and/or B lymphocytes of viruric patients 3 months after renal transplantation. The virus was recovered from a small percentage of T cells (3 per 10(6) cells), but not from B cells. Thus it would appear that virus infection of T cells cannot explain the depressed cell-mediated immunity observed in these transplant patients unless a specific regulatory subset of T cells is infected. However, the isolation of CMV from T cells implies that they must be considered as one of the possible cells for virus persistence in vivo. PMID- 6274986 TI - 'Benign' fibrous histiocytoma of the trachea. AB - Fibro-histiocytic neoplasms are uncommon in the respiratory tract. This paper presents a clinical and histological description of a case of fibrous histiocytoma at the level of the third tracheal ring. This cases shares some features with the four cases previously described, viz., the growth occurs mainly in young adults; recurrences are common; and all have shown similar benign histological patterns. Ultrastructural investigations confirmed the fibro histiocytic nature of the neoplasm. PMID- 6274987 TI - Side chain hydroxylation of C27-steroids and vitamin D3 by a cytochrome P-450 enzyme system isolated from human liver mitochondria. AB - The present study was undertaken to obtain information on the involvement of cytochrome P-450 in the 26-hydroxylation on bile acid intermediates and in the 25 hydroxylation of vitamin D3 in human liver mitochondria. Cytochrome P-450 was solubilized from human liver mitochondria and purified two times to a specific content of 0.125 nmol per mg protein. Furthermore, a ferredoxin was isolated from the mitochondria and partly purified. This iron-sulfur protein had properties similar to bovine adrenal ferredoxin. A mitochondrial NADPH-ferredoxin reductase was also isolated and purified to homogeneity. This enzyme was a flavoprotein with properties very similar to the bovine adrenal NADPH-ferredoxin reductase. The cytochrome P-450 preparation catalyzed 26-hydroxylation of C27-steroids and 25-hydroxylation of vitamin D3 when reconstructed with NADPH, the ferredoxin and the ferredoxin reductase. With different substrates the following turnover numbers (nmol product X nmol P-450(-1) X min-1) were found: cholesterol, 8; 5 cholestene-3 beta, 7 alpha-diol, 10; 7 alpha-hydroxy-4-cholesten-3-one, 23; 7 alpha, 12 alpha-dihydroxy-4-cholesten-3-one, 27; 5 beta-cholestane-3 alpha, 7 alpha-diol, 28; 5 beta-cholestane-3 alpha, 7 alpha, 12 alpha-triol, 41; and vitamin D3, 0.16. The hydroxylation reactions were inhibited by CO and metyrapone. The human liver mitochondrial ferredoxin and ferredoxin reductase could be replaced by adrenal ferredoxin and adrenal ferredoxin reductase without reduction of activity, but they could not be replaced by microsomal NADPH cytochrome P-450 reductase. It is concluded that human liver mitochondria contain cytochrome P-450 involved in the oxidation of the side chain of C27-steroids and vitamin D3. PMID- 6274988 TI - Burkitt's lymphoma. AB - Clinical and epidemiologic features of Burkitt's lymphoma are reviewed. Epidemiologic studies suggest that simultaneous infection with Epstein-Barr (E-B) virus and malaria may be involved as etiologic agents. On the other hand we have found that in the Amazon region of Brazil and Peru both malaria and E-B virus infection is common among children, yet Burkitt's lymphoma is rare. The possibility exists that other concomitant etiologic agents and genetic factors are also involved. Several investigators suggested the possible involvement of Reo 3 virus. We have found antibodies against Yaba virus. A laboratory worker who accidentally inoculated himself with Yaba virus developed a histiocytoma which when inoculated into Asiatic monkeys produced typical Yaba tumors. This was the first case that Koch's postulates were fulfilled in a virus induced neoplasm in man. Therapeutically, the best clinical results were obtained in those patients who were treated with small doses of cyclophosphamide. On the basis of somewhat inadequate follow-up studies, it is estimated that "five year cures" were obtained in about 10% of the patients. PMID- 6274989 TI - Behavioral and electrocortical effects of beta-endorphin after intracerebral infusion in rats. AB - In rats with cannulae chronically implanted into specific areas of the brain, the effects of beta-endorphin were studied on behavior and electrocortical activity. It has been shown that analgesia and catatonia were evoked after infusing beta endorphin into the III cerebral ventricle or into the hypothalamus. On the contrary, beta-endorphin microinjected into the caudate nucleus or into the substantia nigra produced an intense pattern of stereotyped movements, occasional contralateral circling, contralateral myoclonic jerks and asymmetric posture. Stereotyped gnawing, grooming and "wet-dog" syndrome preceded catatonia after intrahypothalamic administration of beta-endorphin. Bilateral (III ventricle) or ipsilateral (hypothalamus, caudate nucleus and substantia nigra) high voltage electrocortical spikes and other ECoG pathological changes accompanied by motor disorders (stereotypies, myoclonic jerks) or without any overt behavioral change were constantly observed. Behavioral and electrocortical changes evoked by beta endorphin were long-lasting and rapidly reversed by the specific opiate antagonist, naloxone. PMID- 6274990 TI - Oat cell carcinoma-the importance of the team approach. PMID- 6274991 TI - Metastatic neonatal Wilms' tumor, A case autopsy report with review of th literature. PMID- 6274992 TI - Receptor activities for low-density lipoprotein and acetylated low-density lipoprotein in a mouse macrophage cell line (IC21) and in human monocyte-derived macrophages. AB - IC21 macrophages, a permanent culture of a line of cells derived from a single colony of mouse peritoneal macrophages transformed with simian virus 40, demonstrate most of the characteristics of lipoprotein metabolism that have been described for primary cultures of rodent or canine peritoneal macrophages. IC21 macrophages have low but demonstrable low-density lipoprotein (LDL) receptor activity. They actively degrade acetylated LDL (AcLDL), which has a negative charge and is not recognized by the LDL receptor. Incubation of IC21 macrophages with human lipoprotein-depleted serum leads to a marked increase in cholesterol synthesis, as measured by incorporation of labeled acetate into sterols. Sterol synthesis is inhibited by further incubation with AcLDL; incubation with LDL also decreases cholesterol synthesis with an accumulation of radioactivity from acetate in sterol intermediates, which indicates that some uptake of LDL occurs. Incubation with AcLDL but not LDL leads to a marked stimulation of cholesterol esterification, as measured by labeled oleic acid incorporation into cholesteryl esters, and a concomitant increase in cellular cholesteryl ester content. IC21 macrophages as compared with human monocyte-derived macrophages are shown to have marked difference in their abilities to degrade native LDL and AcLDL. Human monocyte-derived macrophages degrade LDL at low concentrations at a rate sevenfold greater than do IC21 macrophages. The rate of cholesteryl ester synthesis after LDL receptor induction and incubation with LDL increases linearly with LDL concentration in HMD macrophages, but no increase was found in similarly incubated IC21 macrophages. IC21 macrophages degrade AcLDL at a rate two- to fourfold greater than do human monocyte-derived macrophages. PMID- 6274993 TI - Detection of the human T cell lymphoma virus p19 in cells of some patients with cutaneous T cell lymphoma and leukemia using a monoclonal antibody. AB - A monoclonal antibody specific for the internal p19 protein of a type-C retrovirus (HTLV) isolated from human neoplastic T cells has been developed. Its specificity has been shown by radioimmune precipitation and by affinity chromatography of iodinated HTLV proteins. By indirect immune fluorescence this antibody recognizes only HTLV-producing cells. Examination of cells from patients with cutaneous T cell lymphomas and leukemias and with other types of lymphomas and leukemias indicated that HTLV p19 expression is rare. The monoclonal antibody will be useful in determining the natural reservoir of HTLV, possibly in a subset of mature T cell neoplasias. PMID- 6274994 TI - Mitochondrial N-formylmethionyl proteins as chemoattractants for neutrophils. AB - Mitochondria synthesize several hydrophobic proteins. Like bacteria, mitochondria initiate protein synthesis with an N-formylmethionine residue. Because N formylmethionyl peptides have been found to be chemotactic for polymorphonuclear leukocytes (PMN), mitochondria isolated from cultured human cells and purified bovine mitochondrial proteins were tested for PMN chemotactic activity in vitro. Nondisrupted mitochondria were not chemotactic. However, intact mitochondria that had been incubated with a lysosomal lysate did stimulate PMN migration. Antibodies directed against two mitochondrial enzymes, cytochrome oxidase and ATPase, (both of which contain mitochondrially synthesized subunits) but not anti C3 or anti-C5 decreased mitochondrially derived chemotactic activity. In addition, purified bovine mitochondrial N-formylmethionyl proteins stimulated PMN migration in vitro, whereas nonformylated mitochondrial proteins did not. Furthermore, the chemotactic activity of purified mitochondrial proteins and disrupted mitochondria was decreased by the formyl peptide antagonist butyloxycarbonyl-phenylalanine-leucine-phenylalanine-leucine-phenylalanine. Finally, disrupted mitochondria and purified mitochondrial proteins stimulated PMN-directed migration (chemotaxis), according to accepted criteria. In addition to other chemotactic factors, release of N-formylmethionyl proteins from mitochondria at sites of tissue damage, may play a role in the accumulation of inflammatory cells at these sites. PMID- 6274995 TI - The collagenase of Entamoeba histolytica. AB - The present work was designed to investigate the capacity of trophozoites of Entamoeba histolytica to adhere to and digest human collagen types I and III in vitro. The time-course of binding of ameba to both human collagen types I and III was similar. However, the kinetics of detachment were different for each collagen type. Trophozoites of E. histolytica cultured on heat-reconstituted type I collagen gels produced a well-defined area of lysis. Quantitative studies using 14C-labeled collagen revealed that after 24 h of incubation, Entamoeba digested three and a half times more type I than type III collagen, thus suggesting the presence of a collagenase with higher specificity for type I collagen. This activity was optimum with trophozoites harvested after 42 h in culture (1.5 X 10(5) trophozoites/ml). The digestion of type I collagen was a function of the number of trophozoites, and was inhibited by EDTA, L-cysteine, and serum, but not by soybean trypsin inhibitor, phenylmethanesulfonyl fluoride, or N-ethylmaleimide (NEM). Electrophoretic analysis of the type I collagen fragments revealed three main classes of polypeptides of 75,000, 50,000, and 25,000 daltons. Subsequent proteolysis of these collagen fragments was probably carried out by other proteases derived from trophozoites. This activity was inhibited with 10 mM NEM. Collagenase activity appeared to be located at the plasma membrane and direct contact of the ameba with the substrate is required for collagen digestion. The results suggest that collagenase activity of E. histolytica may play an important role in tissue invasion. PMID- 6274996 TI - A note on the inhibition of in vitro meiotic maturation of mammalian oocytes by dibutyryl cyclic AMP. AB - Cytogenetic data are presented which indicate species differences in the response of oocytes maturing in vitro in the presence of dibutyryl cyclic AMP (dbcAMP). Inhibition of ewe oocyte maturation was noted at germinal vesicle, MI, and MII in the presence of 25-750 microgram/ml of dbcAMP. Cow oocytes were primarily affected at MI and MII by similar concentrations, and only 750 microgram/ml delayed golden hamster oocytes at these stages. These findings highlight the variability between the responses of the oocytes of these mammals and the rat, mouse, or pig. PMID- 6274997 TI - [Surface ultrastructure of the pulmonary alveolar region. Part 2. Common lung cancer cells]. PMID- 6274998 TI - [Cylindroma in otorhinolaryngology]. PMID- 6274999 TI - Partial replacement of succinate dehydrogenase function by phage- and plasmid specified fumarate reductase in Escherichia coli. AB - Phages capable of transducing succinate dehydrogenase mutants (sdh) of Escherichia coli were isolated from pools of artificially constructed recombinant lambda phages using a selective casein digest medium. These phages produced characteristically dense turbid plaques, and as prophages they increased the aerobic growth efficiencies of sdh mutants on complex media but were unable to promote growth with succinate as sole carbon and energy source (an essential feature of sdh + strains). The phages were identified as fumarate reductase transducing phages (lambda frdA) by the presence of a characteristic 4.9 kilobase pairs R.HindIII fragment of bacterial DNA, the expression of a polypeptide with a relative molecular mass of 72,000 (the frdA gene product) and by comparing their transducing activities with authentic lambda frdA phages. In parallel studies a strain containing a ColE1-frd hybrid plasmid (pGS1 = pLC16.43) was characterized. Transfer of pGS1 to sdh mutants was accompanied by increased aerobic growth efficiencies on complex media and the ability to utilize succinate as sole carbon and energy source. It was concluded that fumarate reductase can replace succinate dehydrogenase but the extent of the reversal of the sdh lesion depends on frd gene dosage and the titration of the repressor which normally prevents aerobic synthesis of the reductase. PMID- 6275000 TI - Structure of an experimentally evolved gene duplication encoding ribitol dehydrogenase in a mutant of Klebsiella aerogenes. AB - We have previously described a system of experimental evolution in which many of the mutants of Klebsiella aerogenes selected for faster growth on xylitol ('evolvants') synthesized elevated levels of ribitol dehydrogenase and have presented genetic evidence implicating gene duplication in the enzyme superproduction in some of the evolvants. Here we describe a physical approach to the screening for gene duplications and subsequent structure determination. Nick translated, cloned ribitol operon (rbt) DNA was used as a hybridization probe to identify fragments containing rbt operon sequences in restriction digests of total bacterial DNA. Whilst several of the evolvants probably harbour duplications spanning the entire rbt operon, one of the spontaneously arising evolvants (strain A3) was shown to harbour a small (5.8 kilobase pairs) direct DNA repeat which encodes the dehydrogenase (but not the kinase) of the closely linked D-arabitol operon as well as the dehydrogenase (but not the kinase) of the rbt operon. The hybridization data suggest that there are 4 to 5 copies of the repeat arranged contiguously on the chromosome. The genetic instability of strain A3, the rbt fragment hybridization pattern of an A3 segregant and the activities of the pentitol catabolic enzymes in A3 are all consistent with the proposed gene duplication structure. PMID- 6275001 TI - Characterization of 3': 5' -cyclic AMP phosphodiesterase in Klebsiella aerogenes and its role in substrate-accelerated death. AB - Cyclic AMP phosphodiesterase in Klebsiella aerogenes is a soluble cytoplasmic enzyme with an apparent Km of 0.9 mM and a pH optimum of 7.0. It was inhibited by EDTA, Mg2+ and other metal ions. The enzyme activity was inhibited or activated by some nucleotides but not by any metabolite except pyruvate. It was inhibited by the methylxanthines, caffeine, theophylline and methylisobutylxanthine. During starvation or substrate-accelerated death, the enzyme activity remained essentially constant. It is postulated that during substrate-accelerated death the enzyme acts as a drain on the cellular cyclic AMP levels. The cyclic nucleotide concentrations during substrate-accelerated death are proposed to be controlled directly by adenylate cyclase. PMID- 6275002 TI - Molecular cloning in plasmid pBR322 giving altered expression of the tetracycline resistance gene. AB - The two HindIII fragments of polyoma virus DNA were cloned in the HindIII site of plasmid pBR322, a site located in the RNA polymerase promoter involved in the expression of tetracycline resistance. Although insertion of foreign DNA into this site did not always result in the complete loss of tetracycline resistance, Escherichia coli K12 strain chi 1776 harbouring recombinant plasmids exhibited reduced growth properties in liquid culture with tetracycline and could easily be differentiated from bacteria transformed by non-recombinant plasmids. The formation of plasmid multimers increased the resistance to tetracycline at the level of the induction period, presumably as a result of a gene dosage effect. PMID- 6275003 TI - Involvement of adenosine 3':5'-cyclic monophosphate in the germination of blastospores of Candida albicans. AB - The germination of blastospores of Candida albicans is accompanied by a rise in the intracellular concentration of adenosine 3':5'-cyclic monophosphate (cyclic AMP). Germination was induced either by peptides isolated from seminal plasma or by an amino acid mixture, and both germination and the rise in cyclic AMP required a temperature of 37 degrees C. The rise occurred during the first hour of incubation, but full germination required a temperature of 37 degrees C for 4 h. Germination and the rise in cyclic AMP, in the presence of suboptimal concentrations of inducers, were stimulated by theophylline. Pre-incubation of cells with dithiothreitol in the absence of inducers inhibited subsequent germination but not the rise in cyclic AMP. Germination and the rise in cyclic AMP were inhibited if dithiothreitol or N-succinimidyl-3-(4 hydroxyphenyl)propionate was present during the induction period. PMID- 6275004 TI - Escherichia coli 5'-nucleotidase: purification, properties and its release by osmotic shock. AB - Escherichia coli 5'-nucleotidase was purified to apparent homogeneity as judged by polyacrylamide gel electrophoresis. Its molecular weight was estimated by sodium dodecyl sulphate-polyacrylamide gel electrophoresis, by gel exclusion chromatography, and by membrane filtration; values of 66 000, 48 500, and 15 000 to 30 000, respectively, were obtained. The enzyme was completely released from bacteria by osmotic shock treatment. The apparently anomalous behaviour of 5' nucleotidase in terms of the molecular sieving hypothesis for the release of enzymes by osmotic shock proposed by Smith & Wyatt (1974) and extended by Broad & Smith (1979) is discussed. PMID- 6275005 TI - Hybrid plasmids containing the citrate synthase gene (gltA) of Escherichia coli K12. AB - The Clarke-Carbon colony bank containing ColE1-Escherichia coli hybrid plasmids was screened by conjugation for complementation of the citrate synthase lesion of a gltA mutant. Three ColE1-gltA+ plasmids were identified: pLC26-17 (16.3 kilobase pairs), pLC27-18 (16.35 kb) and pLC31-28 (26.0 kb). The citrate synthase activities of strains containing the hybrid plasmids were amplified 3- to 10 fold. Genetic studies indicated that the smaller plasmids may contain at least part of the succinate dehydrogenase gene (sdh). A physical map of a 19.4 kb region of the E. coli chromosome containing the citrate synthase gene (gltA) was constructed by restriction analysis with the isolated plasmids. The relative positions of 20 restriction sites were defined and a region (3.1 kb) containing the gltA+ gene was identified in the segment common to all three plasmids. PMID- 6275006 TI - Respiratory biogenesis during the cell cycle of aerobically grown Escherichia coli K12. The accumulation of iron-sulphur clusters and their orientation in the membrane. AB - The magnitudes of four signals detectable in intact cells by electron paramagnetic resonance spectroscopy, and assigned to membrane-associated protein, increase continuously during the cell cycle of Escherichia coli K12. Studies on membrane multilayers prepared from cells separated according to their age in the cycle suggest that the orientation within the membrane of the ferredoxin-type signal is invariant throughout the cycle. PMID- 6275007 TI - Sequence relationships between plasmids carrying genes for lactose utilization. AB - Southern hybridization experiments carried out between pSC101::Tn951 DNA and lambda dlac DNA allowed the location and orientation of the lac operon within the transposon to be deduced. The same method was used to detect Tn951 on Lac plasmids from 11 independent isolates from three continents. None of these plasmids was found to carry an entire Tn951 sequence but they all contained lac genes homologous to the lac genes of Tn951. The lac operon of one of these plasmids was bordered by a sequence homologous to that found at the left-hand side of Tn951. It is concluded that the lac determinants of the Lac plasmids analysed and of Tn951 have evolved from a common ancestor but that the distribution of these determinants cannot be attributed to a spread of the transposon Tn951. PMID- 6275008 TI - Plasmids in epidermolytic strains of Staphylococcus aureus. AB - Thirty-four epidermolytic toxin-producing strains of Staphylococcus aureus obtained from a variety of sources were screened for the presence of plasmid DNA. All serotype ii toxin producers harboured a large 42 kilobase pairs (kb) plasmid. Elimination of these plasmids resulted in the simultaneous loss of a bacteriocin determinant (Bac+) and type ii toxin production (Toxii+). Some strains producing serotype i toxin (Toxi+) contained similar 42 kb plasmids. Elimination of these plasmids resulted in the loss of only bacteriocin production. Strains producing both toxin serotypes readily lost Toxii+ and Bac+, which were carried on the same plasmid, but Toxi+ could not be eliminated. Thus Toxi+ was probably chromosomally determined, while Toxii+ was a plasmid-encoded marker. In some strains cadmium resistance was also linked to the 42 kb plasmid. The 42 kb plasmids from seven strains with different phenotypes were analysed with restriction endonucleases EcoRI and HindIII. The plasmids shared 19 of 22 HindIII fragments indicating that they are closely related to each other. PMID- 6275009 TI - Porin from the outer membrane of Escherichia coli: immunological characterization of native and heat-dissociated forms. AB - Antisera against porin oligomers isolated from the outer membrane of Escherichia coli O26K60 and against porin monomers from the same bacterial strain were elicited in rabbits by intramuscular administration with Freund's complete adjuvant. Antibodies against native porin oligomers reacted strongly with porin oligomers, as revealed by sodium dodecyl sulphate-polyacrylamide gel immunoperoxidase (SGIP) analysis, the enzyme-linked immunosorbent assay (ELISA) and immunodiffusion, but showed no significant reaction with denatured monomers. The antibodies were completely absorbed by the intact outer membrane peptidoglycan complex, which suggests that they were directed against antigenic determinants expressed on the outside of the intact outer membrane. Antibodies directed against denatured porin monomers reacted strongly with monomers in all tests but reacted only very weakly with porin oligomers. They were not absorbed by the native porin situated in the intact outer membrane. This indicates that the major antigenic determinants of the denatured porin monomer are hardly related to those of the native trimer situated in the intact outer membrane. The antigenic determinants of the denatured monomer seem to become fully expressed only after dissociation and denaturation of the porin. It is concluded that the immunological relationship of denatured porin monomers derived from many strains of E. coli and other Enterobacteriaceae which was reported in previous studies may not indicate that native porin trimers of these strains are also related. PMID- 6275010 TI - DNA methylation--a regulatory signal in eukaryotic gene expression. PMID- 6275011 TI - Mapping temperature-sensitive mutants of vesicular stomatitis virus by RNA heteroduplex formation. AB - Duplex RNA molecules made by hybridization of virion and mRNA of vesicular stomatitis virus (VSV) were digested with ribonuclease and separated into five size classes, each containing the gene and the mRNA for one of the VSV proteins. Denaturation of the duplexes yielded full size mRNA lacking poly(A) tails. Utilizing duplex formation between the RNAs from VSV temperature-sensitive (ts) mutants and their revertants and subsequent RNase digestion under varying salt conditions, specific cleavages within a certain duplex were seen for representative mutants from complementation groups, III, IV and V. Specific cleavages were not seen for a group II mutant. From these results gene assignments cannot be made for group II; equivocal assignments are made for group III and clear assignments made for group IV and V. The assignment for the group V mutants, however, does not conform to expectations. Nevertheless, from these studies and other published ones, there is the suggestion that interactions may exist between the gene products of complementation groups II and V during VSV transcription and morphogenesis. These results also support the lack of transcriptional splicing for VSV mRNAs. PMID- 6275013 TI - Neonatal infection with mouse thymic virus: spleen and lymph node necrosis. AB - Mouse thymic virus (TA) is a naturally occurring herpesvirus of laboratory and wild mice, which produces massive thymic necrosis when inoculated into newborn mice. Our histopathological study showed necrosis not only of the thymus but also of the spleen and lymph nodes which was noticeable by day 7 and complete by day 14. Both spleen and lymph nodes regenerated to an almost normal histological pattern by day 70. The results show that TA infects multiple lymphoid tissues causing massive necrosis in all, and is not limited to a single site, the thymus. TA infection was found to be a persistent herpesvirus infection in both the lymph nodes and spleen. During the period of acute infection, as necrosis increased, the response of cell suspensions of lymph nodes to the T cell mitogens concanavalin A and phytohaemagglutinin was virtually non-existent. Activity returned to normal as the histological repair progressed. PMID- 6275012 TI - Antibody-dependent enhancement of plaque formation on cell lines of macrophage origin - a sensitive assay for antiviral antibody. AB - An assay for antiviral antibody based on antibody-dependent plaque enhancement (ADPE) in the macrophage cell line P388D1 is described which is as sensitive as, or more sensitive than a radioimmune assay. The method is applicable to a range of Togaviridae and Bunyaviridae although not to Picornaviridae and Herpesviridae. The variables affecting the assay are investigated. PMID- 6275014 TI - Human cytomegalovirus-associated DNA polymerase and protein kinase activities. AB - Human cytomegalovirus (HCMV), purified exclusively from the extracellular media, contained a DNA polymerase activity in addition to a protein kinase activity. The DNA polymerase expressed its maximum activity in the presence of 5 to 10 mM MgCl2. The enzyme was able to use effectively activated calf thymus DNA, poly(dA).oligo(dT)12--18 and poly(dC).oligo(dG)12--18 as the template primers. The DNA polymerizing activity was eluted with 0.18 to 0.2 M-KCl from a phosphocellulose column. It was relatively resistant to phosphonoacetic acid inhibition even at a high concentration of 100 micrograms/ml with activated calf thymus DNA as the template primer, but the DNA polymerase activity was totally suppressed at this concentration when poly(dA).oligo(dT)12--18 was used as the template primer. The enzyme activity was inhibited by ammonium sulphate at 0.01 to 0.3 M with either activated calf thymus DNA or poly(dA).oligo(dT)12--18 as the template primer. The protein kinase has maximum activity in the presence of 10 to 20 mM-MgCl2, and preferred virion proteins as phospho-acceptor to protamine sulphate. Histone, caesin and bovine serum albumin (BSA) were found to be poor substrates. The phosphorylated protein pattern of the in vivo [32P]orthophosphate labelled virions was not identical to that of the in vitro phosphorylated Nonidet P40-dissociated virions, although seven phosphorylated polypeptides did co migrate in SDS--polyacrylamide gel electrophoresis (SDS--PAGE). Procedures known to solubilize virions showed that the DNA polymerase and protein kinase were internal components of the virion. PMID- 6275015 TI - Proteus mirabilis converting phage 5006Mpa has an oversized genome. AB - Proteus mirabilis phage 5006Mpa is a converting variant for ampicillin resistance of phage 5006M. We show here that the ampicillin resistance marker of transposon Tn1 is located on a 9.8% insertion with respect to the wild-type phage genome. This renders the 5006Mpa genome 5% oversized, albeit without loss of wild-type genetic material from the phage population. PMID- 6275016 TI - Flow cytometric analysis of DNA content of mouse liver cells following in vivo infection by human adenovirus type 5. AB - Human adenovirus type 5 caused acute hepatocellular damage when injected intravenously into C57B1/6 mice. Early protein (P) and late virus protein (V) antigen staining by immunofluorescence was located principally, if not exclusively, in hepatocytes. Autoradiography following incorporation of tritiated thymidine into Ad5-infected liver cells in vivo indicated that DNA synthesis was initiated in hepatocytes. Flow cytometry showed that cells with a 4n DNA content prior to infection were highly susceptible to infection, with 75% being lost from the liver cell population by 5 days post-infection. This observation was correlated with the observed pathology during the course of the infection. PMID- 6275017 TI - Mechanism of early and late polykaryocytosis induced by the bovine leukaemia virus. AB - Syncytia formation induced by the bovine leukaemia virus (BLV) has been classified as 'early' or 'late' polykaryocytosis. Early polykaryocytosis arises in the first 24 h in mixed cultures of BLV-infected cells with indicator cells. Late polykaryocytosis is observed 4 to 8 days after infection of sensitive cells with cell-free infectious BLV. Our results demonstrate that the two phenomena proceed from different mechanisms. Late polykaryocytosis results from an active process dependent on the integrity of the virus genome. In contrast, early polykaryocytosis is a passive process which does not require any de novo virus synthesis but is dependent on the presence of virus proteins in inducer cells. PMID- 6275018 TI - The interaction of encephalomyocarditis virus with its erythrocyte receptor on affinity chromatography columns. AB - Glycophorin, the major sialoglycoprotein in the human erythrocyte surface membrane, can serve as a red cell receptor for both wheat-germ agglutinin (WGA) and encephalomyocarditis (EMC) virus since glycophorin bound to WGA--Sepharose can at the same time bind EMC virus. In contrast, glycophorin bound to WGA- Sepharose cannot bind EMC virus in the presence of SDS. The evidence suggests that virus binding to glycophorin-WGA--Sepharose occurred in the absence of SDS because glycophorin was present in aggregated complexes which were large enough either to accommodate both EMC virus and WGA at the same time, or alternatively to provide sufficient attachment sites for multivalent binding of virions. PMID- 6275019 TI - Analysis of deoxycytidine (dC) deaminase activity in herpes simplex virus infected or HSV TK-transformed cells: association with mycoplasma contamination but not with virus infection. AB - Deoxycytidine (dC) deaminase activity has been previously reported to be induced in herpes simplex virus (HSV)-infected cells (Chan, 1977). In contrast, we report here that HSV infection of either hamster cells naturally deficient in this enzyme activity or mouse cells containing a low level of activity never resulted in appearance of stimulation of dC deaminase, whereas thymidine kinase (TK) was always induced. Surprisingly, dC deaminase activity, which differed by electrophoretic mobility from the mouse or human cell enzyme, was discovered in some cells selected for the presence of HSV TK after infection with u.v. irradiated HSV. Evidence is presented which suggests that the appearance of this new enzyme was not due to the presence of virus genes but rather to mycoplasma contamination. PMID- 6275020 TI - Effect of 2-deoxy-D-glucose on cytomegalovirus-induced DNA synthesis in human fibroblasts. AB - 2-Deoxy-D-glucose (dGlc) was found to selectively inhibit virus DNA synthesis in human embryonic lung cells infected with human cytomegalovirus (HCMV). The effective concentration of dGlc was approx. 10-fold higher in culture medium containing glucose instead of sodium pyruvate. This inhibitory action of dGlc was fully reversible following replacement of the inhibitor medium by fresh medium after a 48 h treatment of infected cells. Virus DNA synthesis could be selectively inhibited by addition of dGlc even after initiation of HCMV DNA replication. In contrast, virus DNA synthesis in herpes simplex virus-infected cells was insensitive to dGlc. The drug was found to deplete HCMV-infected cells of uridine triphosphate and caused a progressive reduction of uridine incorporation into RNA. To substantiate a possible interference by dGlc with the expression and/or function of virus-induced, chromatin-associated factors essential for virus DNA replication, DNA synthesis of chromatin preparations from dGlc-treated, HCMV-infected cells was analysed. In contrast to preparations of untreated or phosphonoacetic acid (PAA)-treated, HCMV-infected cells, those of dGlc-treated cells lacked significant in vitro DNA-synthesizing activity; virus DNA was not synthesized by these preparations. Tunicamycin in the presence of low concentrations of dimethyl sulphoxide was also found to be effective in abolishing HCMV-induced DNA replication. It is thus suggested that dGlc interferes with the function of an 'early' chromatin-associated glycoprotein essential for virus DNA synthesis. PMID- 6275021 TI - Host cell-dependent differences in the oligosaccharide moieties of the VSV G protein. AB - The oligosaccharide moieties of vesicular stomatitis virus glycoprotein from virus grown in four different cell lines have been characterized by sequential enzymic degradation followed by ion-exchange chromatography and analytical gel filtration. Whilst the same two peptide sites are glycosylated in all cell lines, the extent of sialylation of the oligosaccharides is, however, a function of the cell line in which the virus is produced. Using specific purified glycosidases for sequential degradation of glycopeptides obtained after Pronase digestion, the oligosaccharide structures from the different host cell lines appear similar. However, differential sensitivity of the glycopeptides to treatment with a partially purified mixture of endo- and exoglycosidases indicates that the oligosaccharide structures are not identical. PMID- 6275022 TI - Early interaction between mouse hepatitis virus 3 and cells. AB - The interaction between mouse hepatitis virus 3 (MHV3) and cells was studied in order to investigate whether or not early events occurring after infection could be involved in the difference in virus replication seen between mouse strains with different genetic sensitivities to MHV3 infection. Kinetic data showed that MHV3 uptake by both macrophages and L cells was time- and temperature-dependent. In addition, treatment of cells with cytochalasin B or prostaglandin E1, prior to virus infection, resulted in a strong inhibition of sheep red blood cell phagocytosis without any effect on MHV3 uptake. Similar uptake of radiolabelled MHV3 was shown by whole spleen cells, purified T lymphocytes and thymocytes. Furthermore, no difference in 3H-labelled MHV3 uptake was seen between macrophages originating from resistant A/J mice, semi-susceptible (C57Bl/6 x A/J)F1 and susceptible animals. These results indicate, therefore, that genetically related in vivo sensitivity toward MHV3 infection is not related to differential uptake of virus by cells. PMID- 6275023 TI - Variety of endogenous proviruses in the genomes of chickens of different breeds. AB - We have compared endogenous proviruses in DNA of chickens of 11 breeds by means of Southern's technique. Many of the endogenous virus loci found were missing from the genomes of the extensively studied white leghorn chickens, although some of the proviruses and especially ev-1 appeared to be widely spread among different chickens. Most of the proviruses were similar to Rous-associated virus (RAV-O) as judged by the EcoRI digestion patterns. Several genetically different proviruses were found in the DNA of 28 brown leghorns. They contain neither ev-1 provirus in their genome, nor any other one common for all individuals. All four Italian partridge-coloured chickens examined appeared to be free from ALV-related sequences in their DNA showing the possibility of normal life without known endogenous proviruses. Possible causes of inter-breed differences of chicken endogenous proviruses are discussed. PMID- 6275024 TI - Detection of hepatitis B virus-specific DNA in the genomes of human hepatocellular carcinoma and liver cirrhosis tissues. AB - Hepatitis B virus-related DNA was detected in the chromosomal DNA of three out of seven hepatocellular carcinomas and two out of five cirrhosis samples examined, by means of the blot-hybridization technique, described by Southern (1975). The integration patterns were not identical but some similarities raise the question of whether there are some preferred sites of viral integration. PMID- 6275025 TI - Comparison of bluetongue type 20 with certain viruses of the bluetongue and Eubenangee serological groups of orbiviruses. AB - The genome of bluetongue virus type 20 consists of 10 segments of double-stranded RNA each of which contains unique sequences as determined by oligonucleotide mapping. The 10 polypeptide products of the virus genome were detected in virus infected cells, and in pulse--chase experiments there was no secondary cleavage of the primary gene products. Using stringent conditions for RNA--RNA reassociation, no significant homology could be detected between the genomes of bluetongue type 20 isolated in Australia and representative serotypes isolated in other geographic regions. The results suggest sequence divergence between geographically isolated viruses and not the recent introduction of a bluetongue virus into Australia. PMID- 6275026 TI - Persistence and expression of Marek's disease virus DNA in tumour cells and peripheral nerves studied by in situ hybridization. AB - We have used cloned fragments of Marek's disease virus (MDV) DNA and in situ hybridization to search for virus DNA and study its expression in infected chick embryo fibroblasts (CEF), lymphoblastoid cell lines, tumours and neural lesions. DNA from the HPRS 16/att strain of MDV was cleaved with EcoRI endonuclease and several fragments were cloned in Escherichia coli using the vector PBR322. Seven fragments ranging in size from 2.6 to 11 kbp representing approx. 25% of the MDV genome were labelled in vitro and annealed to EcoRI digests of DNA from infected cells and tumours following separation and transfer according to the Southern blotting procedure. Most of the selected MDV DNA fragments hybridized to fragments of corresponding sizes in EcoRI digests of DNA from cell lines and tumours and failed to hybridize to digests of uninfected chick cell DNA. In situ hydridization using 3H-labelled DNA with specific activity of 10(8) d/min/microgram as probe showed intranuclear MDV DNA in infected CEF, in every cell of two lymphoblastoid cell lines and in the majority of infiltrating or proliferating lymphoid cells found in type 'A' lesions of grossly enlarged peripheral nerves. Both intranuclear and cytoplasmic RNA were detected in cells that contained virus DNA. However, comparatively little virus RNA appears to be transcribed in cell lines and in infected tissues from the regions of virus DNA (25% of genome) used as probe in this study. Our results favour the hypothesis that the accumulation of lymphoid cells in nerves is not the result of an inflammatory response to infected nerve cells but is rather the consequence of proliferating transformed cells. PMID- 6275027 TI - Complementation between phosphonoacetic acid-resistant and -sensitive variants of herpes simplex viruses: evidence for an oligomeric protein with restricted intracellular diffusion as the determinant of resistance and sensitivity. PMID- 6275028 TI - Analysis of the genome of equine herpesvirus type 1: arrangement of cleavage sites for restriction endonucleases EcoRI, BglII and BamHI. AB - The genome of an Australian isolate of equine herpesvirus type 1 (equine abortion virus) has been analysed using the restriction endonucleases EcoRI, BglII and BamHI, and a physical map constructed. Terminal fragments were identified by exonuclease treatments, and linkage of fragments was deduced by a combination of single- and double-digest experiments and cross-blot hybridizations. The genome has a mol. wt. of 100 x 10(6) and is comprised of a short unique region bounded by repetitive sequences, which is present in both orientations in approximately equal amounts in the DNA population, and a long unique region existing in only one orientation. PMID- 6275029 TI - Murine cytomegalovirus particle types in relation to sources of virus and pathogenicity. AB - Murine cytomegalovirus (MCMV) preparations from mouse embryo fibroblasts and from infected salivary glands were purified on potassium tartrate density gradients and examined by electron microscopy. The cell culture virus contained multicapsid enveloped virions which are ruptured easily and account for the excess number of free capsids in these preparations. Salivary gland virus consisted of single capsid enveloped virus and equal numbers of free capsids. Particle types were imperfectly separated on density gradients, but successful separation was achieved by filtration through 220 and 450 nm Millipore membrane filters. Naked capsids were not infectious and could not be rendered infectious by centrifugal adsorption, showing that centrifugal enhancement of MCMV infectivity is not mediated through this mechanism. Although present in excess number in (avirulent) cell culture virus preparations, naked capsids did not interfere with the action of (virulent) salivary gland virus in newborn mice. PMID- 6275030 TI - Determination of the equilibrium dissociation constants and number of glycine binding sites in several areas of the rat central nervous system, using a sodium independent system. AB - Parameters affecting the binding of [3H]glycine to membrane fractions isolated from the cerebral cortex, midbrain, cerebellum, medulla oblongata, and spinal cord of the rat were investigated in a Na+-free medium. A [3H]glycine binding assay was established in which the binding was specific, saturable, pH-sensitive, and reversible. Conditions were chosen in an effort to minimize binding to glycine uptake sites. From data on specific [3H]glycine binding Scatchard plots were prepared and the KD and Bmax values were calculated. Two glycine binding sites (high and low affinity) were identified only in the medulla (KD: 44, 211 nM; Bmax: 361, 1076 fmol/mg protein) and spinal cord (KD: 19, 104 nM; Bmax: 105, 486 fmol/mg protein). The ranges of the KD and Bmax values for the other three areas studied were 59 to 144 nM and 882 to 3401 fmol/mg protein, respectively. When the glycine content of each area, expressed as fmol/neuron, was plotted against the respective KD (high affinity), a negative correlation was found (r = -0.90; p less than 0.05). A similar negative correlation was found between the glycine content and Bmax (r = --0.88; p less than 0.05). Hill plots indicated a slope of essentially 1.0 for all areas. GABA, taurine, strychnine, diazepam, bicuculline, and imipramine had little or no effect on [3H]glycine binding. PMID- 6275031 TI - Glutamic acid decarboxylase and gamma-aminobutyric acid in Huntington's disease fibroblasts and other cultured cells, determined by a [3H]muscimol radioreceptor assay. AB - A sensitive and reproducible [3H]muscimol radioreceptor assay was developed for measuring low levels of both glutamic acid decarboxylase activity and gamma aminobutyric acid. By using this technique, endogenous gamma-aminobutyric acid and glutamic acid decarboxylase activity were detected in two rat neuroblastomas, B35 and B50, a human medulloblastoma cell line, TE671, and cultured human skin fibroblasts. Glutamic acid decarboxylase activities and gamma-aminobutyric acid levels were compared for human skin fibroblasts obtained from patients with Huntington's disease and their controls in a well-controlled, blind study. However, no significant difference was found to either measure between Huntington and control cells. Glutamic acid decarboxylase activity was relatively low in all cell types examined except for the TE671 cells, which had more than four times the activity found in the other cells. This human medulloblastoma cell line appears to be a good model for studying gamma-aminobutyric acid metabolism and the control of glutamic acid decarboxylase expression. PMID- 6275032 TI - [3H]Diazepam displacing activity in human cerebrospinal fluid. AB - Pooled human cerebrospinal fluid was separated by Sephadex G-50 chromatography. The presence of three peaks, A, B and C, was demonstrated by monitoring absorbance at 254 and 280 nm. All peaks showed [3H]diazepam displacing activity in the membrane receptor test. Peak B was further separated on Bio-Gel P-4. At least two major fractions free of salt and GABA in the molecular weight range of approximately 700--3600 were shown to displace [3H]diazepam in the receptor test. This activity was enhanced by a factor of 3 in the presence of 10 microM-GABA. PMID- 6275033 TI - Electroconvulsive shock and reserpine: effects on beta-adrenergic receptors in rat brain. AB - Electroconvulsive shock (ECS) administered once daily for up to 14 days decreases beta-adrenergic receptor binding in the cortex and hippocampus in a time dependent manner. The decrease in binding in the cortex lasts at least 1 week after the last shock. In the striatum, hypothalamus, or cerebellum, 14 days of ECS did not produce significant changes in beta-adrenergic receptor binding. The brain regional pattern of beta-adrenergic receptor changes suggests that repeated ECS affects beta 1-adrenergic receptors in brain regions that receive a noradrenergic innervation activated by ECS. The effects of ECS on neurotransmitter receptor binding appear to be highly selective. Of five receptors in the cortex and three receptors in the hippocampus measured, only beta-adrenergic receptor binding is decreased. Chronic footshock stress does not alter beta-adrenergic receptor binding sites in the cortex, indicating that the effects of ECS are not due to stress alone. The effects of ECS on reserpine induced alterations in beta-adrenergic receptor binding sites were also examined. Ten days of ECS following chronic reserpine injections reverses the increased binding of beta-adrenergic receptors. PMID- 6275034 TI - Inhibition of [3H]GABA binding to postsynaptic receptors in human cerebellar synaptic membranes by carboxyl and amino derivatives of GABA. AB - Fifty synthetic analogues of GABA were tested for their ability ot interact with GABA receptors, using [3H]GABA binding to human cerebellar membranes as an in vitro model. The most active compounds were found to be aliphatic and heterocyclic aminosulphonic acids. Compounds with highly substituted nitrogen atoms were only weakly active unless a long alkyl chain, which can interact with the postsynaptic membrane, was present. It was concluded that a pyramidal nitrogen atom is favoured fro binding of GABA analogues to human cerebellar membranes. PMID- 6275035 TI - Intracellular localization of 2',3'-cyclic nucleotide 3'-phosphodiesterase in a neuronal cell line as examined by immunofluorescence and cell fractionation. AB - The enzyme 2',3'-cyclic nucleotide 3'-phosphodiesterase (CNPase, EC 3.1.4.37) occurs not only in myelin fractions and glial cells, but can also be shown to be present in a CNS cell line of neuronal origin (B104). Direct immunofluorescence microscopy of B104 cells with fluorescein isothiocyanate-conjugated rabbit anti CNPase antibodies shows a discrete and specific intracytoplasmic location of CNPase. Fractionation of the cells was performed by differential centrifugation of a cell homogenate and continuous sucrose density-gradient centrifugation. As monitored by marker enzyme activities, CNPase seems to be associated with endoplasmic reticulum membranes. PMID- 6275036 TI - Immunologic requirements for the adoptive transfer of ectromelia virus meningitis. AB - Intracerebral infection with the Hampstead egg strain of ectromelia virus in mice produced a meningitis with clinical signs and death. A single injection of cyclophosphamide three days after infection delayed the onset of pathology and clinical signs. Adoptive transfer of immune spleen cells into preinfected recipients could induce the meningitis earlier. These donor-immune cells were virus-specific in their action. The Ig- population, deficient in B lymphocytes, could transfer meningitis; but donor-immune cells depleted of T lymphocytes by anti-theta treatment or the Ig+ population, deficient in T-cells, were unsuccessful. Studies with labeled cells showed a preferential accumulation of labeled donor cells in cerebrospinal fluid in the first 24 hours post-transfer, whereas labeled recipient cells accumulated between 24 and 48 hours. The results demonstrated that the adoptive transfer of ectromelia virus meningitis was a cell mediated inflammatory process. PMID- 6275037 TI - The role of the major histocompatibility complex in the adoptive transfer of ectromelia virus meningitis. AB - Adoptive transfer of ectromelia virus meningitis was most efficient when donor immune spleen cells and recipients were compatible in the K region of the H-2 gene complex. Weak responses could be obtained with H-2D region compatibility, but none occurred with H-2I region compatibility. Spleen cells used in adoptive transfers and cells found in cerebrospinal fluid from infected mice were found to have identical H-2-imposed restriction in their in vitro cytotoxic activity. This suggests a significant role for the major histocompatibility complex in the generation of virus-specific T lymphocytes and the recognition of virus-infected tissue in the central nervous system by these cells. PMID- 6275038 TI - Gaucher's disease and glioblastoma multiforme in two siblings: a clinicopathologic study. AB - The clinical and pathologic features of two brothers with the adult form of Gaucher's disease, both of whom developed cerebral glioblastoma multiforme, are presented. Neither patient had a long-standing neurologic disorder, and morphologic evidence of nervous system glucocerebroside storage was not seen. Despite the known increased incidence of malignancy in Gaucher's disease, a review of the literature and of sixty-two Mayo Clinic cases of adult onset Gaucher's disease revealed only one other instance of the association with glioma. The significance of the relationship is unknown. PMID- 6275039 TI - The fine structure of hyaline inclusions in meningioma. AB - Hyaline inclusions of three meningiomas were studied at the ultrastructural level. The inclusions apparently arise as the result of fusion of smaller microvillus-containing intracytoplasmic cysts. The content of the inclusions consists of finely reticulated filamentous material. These findings suggest a secretory differentiation of meningeal cells in these meningiomas. PMID- 6275040 TI - Nonlinear interactions in ganglion cell receptive fields. PMID- 6275041 TI - Tactile spatial resolution. II. Neural representation of Bars, edges, and gratings in monkey primary afferents. PMID- 6275042 TI - Patterns of increased metabolic activity in somatosensory cortex of monkeys Macaca fascicularis, subjected to controlled cutaneous stimulation: a 2 deoxyglucose study. PMID- 6275043 TI - Rheobase, input resistance, and motor-unit type in medial gastrocnemius motoneurons in the cat. PMID- 6275044 TI - Homonymous projection of individual group Ia-fibers to physiologically characterized medial gastrocnemius motoneurons in the cat. PMID- 6275045 TI - gamma-Aminobutyric acid concentration, L-glutamate 1-decarboxylase activity, and properties of the gamma-aminobutyric and postsynaptic receptor in cobalt epilepsy in the rat. AB - Crude mitochondrial synaptosomal (P2) fractions were used to measure L-glutamate 1-decarboxylase (GAD) activity, and crude synaptic membranes were isolated from rat brains and used to determine gamma-aminobutyric acid (GABA) concentration and postsynaptic GABA receptor binding characteristics in rats with cobalt, copper, or glass implanted in right and left cerebral cortices. Copper was employed as a positive metal control because it elicits a morphological profile similar to that of cobalt but is non-epileptogenic. From tissue adjacent to the lesion, GAD activity was assessed by counting trapped 14CO2 liberated from [14C]glutamate and was reduced maximally to 25% of glass controls 7 days following cobalt insult, a period of peak seizure incidence. No reduction in GAD activity was observed 1 or 21 days after cobalt treatment or at any time period in copper-or glass-treated animals. A radioligand [3H]GABA receptor assay was utilized to determine GABA levels, postsynaptic receptor number (Bmax), and the affinity of the postsynaptic receptor for the ligand (KD) in tissue surrounding the lesion. GABA concentration was reduced maximally to 47% of glass controls 7 days following cobalt implantation. Scatchard plot analysis of tissue adjacent to the cobalt lesion revealed a significant increase in apparent receptor density (Bmax) to 200% of glass controls 7 days after bilateraL cobalt implantation (Bmax = 3.97 +/- 0.83 pmol/mg of protein, cobalt versus 1.36 +/- 0.17, glass control). Moreover, at 7 days, no change in kinetic parameters was noted after copper treatment. From days 7 to 21, the density (Bmax) of postsynaptic GABA receptors in cobalt-treated tissue appears to return slowly to glass control values. Results from the present study suggest that degeneration of the GABA pathway in the frontal cortex of the cobalt-epileptic rat occurs and, coupled with the increased Bmax, may represent a "denervation supersensitivity" phenomenon. PMID- 6275046 TI - Extracellular current flow and the site of transduction by vertebrate hair cells. AB - The transduction process of a vertebrate hair cell commences with the application of mechanical stimuli to the hair bundle, a cluster microvillous stereocilia and single axonemal kinocilium. In an effort to determine where within the hair bundle transduction occurs, I have measured extracellular potentials around the hair bundles of mechanically stimulated hair cells from the bullfrog's sacculus. Stimulus-dependent signals up to 17 microV in peak-to-peak amplitude have been found. These appear to be due to the flow of transduction current on the basis of their amplitude, phase, dependence on stimulus size and orientation, proportionality to membrane potential, and sensitivity to an ototoxic antibiotic. The responses are consistently larger near the top of the hair bundle than at its base, suggesting that the transduction apparatus lies at or near the distal ends of the stereocilia. PMID- 6275047 TI - Correlative firing patterns of serotonergic neurons in rat dorsal raphe nucleus. AB - In this study, local neuronal interactions in the midbrain dorsal raphe nucleus (DRN) were analyzed by the use of autocorrelation and cross-correlation histograms. The autocorrelograms of serotonin (5-hydroxytryptamine; 5-HT) containing cells showed an initial trough and subsequent periodicity which corresponded to their regular rhythmic firing pattern. Cross-correlograms revealed that all adjacent 5-HT neurons recorded from single micropipettes displayed complicated patterns of functional interactions. The interactions could be grouped into the following categories: (1) synchronization (interspike intervals less than 10 msec), (2) synchronization and direct inhibition, (3) synchronization and mutual inhibition. In contrast, cross-correlograms generated from adjacent neuronal pairs consisting of one 5-HT cell and one non-5-HT neuron or from non-adjacent 5-HT neuronal pairs usually failed to show functional interactions. The results of the present study are consistent with previous findings of a powerful auto-and mutual regulatory system for 5-HT neurons in the DRN. PMID- 6275048 TI - Immunological, biochemical, ultrastructural, and electrophysiological characteristics of a human glioblastoma-derived cell culture line. AB - This report presents the results of a study using multiple techniques of the established human cell line, LM, which has been developed in culture medium from a patient with a right temporoparietal glioblastoma. This cell line has human subtetraploid karyotype and has several features of a transformed line in culture. These include continuous propagation for 10 years, ability to form tumor nodules when transplanted into immunologically suppressed hamsters, and pleomorphic appearance. Ultrastructurally, it is characterized by multiple nuclei, few actin cables, and numerous surface-membrane microvilli, as well as abundant 9- to 10-nm cytoplasmic filaments. By its immunological reactivity, the line can be shown to contain glial fibrillary acidic protein at low levels, consistent with its glial origin and continued nature. Dibutyryl cyclic adenosine monophosphate (db-cAMP) induces formation of long astrocytic-like processes as well. Its membrane electrical characteristics include a low resting membrane potential and short time constant. Used in a microtiter antiglioma antibody cytotoxicity assay, LM yields a positive reaction to antibodies in the sera of 80% of patients with astrocytomas and only 9% of normal blood-bank donors, suggesting that it shares common antigens with other astrocytic tumor lines. The varied characteristics of this glioblastoma-derived line emphasize the "multiforme" nature on the neoplasm and suggest that for characterization of any such line, multiple parameters are necessary to allow comparison with other long term glioblastoma lines in the literature. The usefulness of the LM line in in vitro cell biological, immunological, chemotherapeutic, and radiobiological studies of gliomas makes such efforts very worthwhile. PMID- 6275049 TI - Right anterior oblique first-pass radionuclide ejection fractions: effects of temporal smoothing and various background corrections. AB - Thirty-seven patients undergoing contrast left ventriculography were studied by first-pass radionuclide angiography (FPRA) in the right anterior oblique view. Ejection fraction (LVEF) was calculated from FPRA using (a) a spatially and temporally varying background correction (BGC) based on a matrix of activity in lung and left atrium and (b) BGC with temporal fluctuation but with no allowance for spatial variations. The two methods were performed on both raw and temporally smoothed data. All four LVEFs correlated well with contrast LVEF (r = 0.90 - 0.94). Absolute values differed significantly from contrast values except for the method using the spatially and temporally varying BGC on smoothed data, which provided the closest overall agreement at all levels of LVEFs, despite occasional large individual variations. The same method on raw data overestimated low LVEFs and the method applying only temporal fluctuation in background underestimated high LVWFs. Allowance for spatial and temporal variations in background is therefore important when first-pass radionuclide angiography is performed in the RAO view. PMID- 6275050 TI - Compton scatter image simulating jugular venous reflux. AB - In a radionuclide cerebral angiographic study, Tc-99m photons from the subclavian vein may scatter in the superficial tissues of the neck and head, resulting in an image simulating the jugular venous reflux. In a scintillation camera peaked at 140 keV with a 20% window, any scattered photons with a scatter angle of less tha 53.5 degrees may be counted in the Tc-99m window. This scatter angle is large enough to allow counting of many secondary photons from Compton collisions in an area quite distant from the radioactive source to be counted, provided the scatter area and source are separated by air. PMID- 6275051 TI - Re:diagnosis of alcoholic cirrhosis with the right-to-left hepatic lobe ratio: concise communication. PMID- 6275052 TI - Tc-99m PPi localization in acute experimental myocardial infarction: application of macro- and microautoradiography. PMID- 6275053 TI - Influence of dietary fibers on fermentation in the human large intestine. AB - Fiber sources were fed to 24 men to evaluate the effect of fiber on large intestinal microflora. The diets were low fiber basal or the basal diet supplemented with coarse bran, fine bran, cabbage, or cellulose (Solka floc). Twelve subjects (Group 1) were fed all fiber sources using a latin square design with 2-week experimental periods followed by a 3-week period on the basal diet. Group 2 made one diet change after the first experimental period, then remained on that diet throughout the experiment; during the last period their fiber intake was double (to approximately 26 g/day). A microbial culture system was utilized with fecal microflora to measure cell wall indigestibility and volatile fatty acid (VFA) production. Volatile fatty acids were also measured in fresh feces. The in vitro substrates were alfalfa, coarse bran, cabbage, or cellulose. In this system, dietary fiber had a significant (P less than .05) effect on resultant cell wall indigestibility and VFA level. Previous diet in group 1 subjects significantly (P less than .05) affected in vitro VFA. In groups 2 significant (P less than .05) period and subject effect were present for the in vitro VFA data. The VFA concentrations in feces of humans were significantly affected by the fiber consumed. PMID- 6275054 TI - Effects of processed rye bran and raw rye bran on glucose metabolism in alloxan diabetic rats. AB - Alloxan diabetic rats (D) and control rats (C) were for 14 days fed a basic diet made from a mixture of rye bread and milk containing either no bran (group O), processed bran (group PB) or raw bran (group B). The D-B animals has a lower urinary glucose excretion (36 +/- 4 mmoles/24 hours) than the D-PB animals (63 +/ 5 mmoles/24 hours), which in their turn had a lower excretion than the D-0 animals (100 +/- 3 mmoles/24 hours). The effects of the two brans on weight, serum insulin levels, and non-fasting blood glucose levels were similar in D-rats as well as in C-rats, with the exception of the lower nonfasting blood glucose level in C-B animals than in C-PB animals. PMID- 6275055 TI - 3-hydroxy-3-methylglutaryl coenzyme A reductase activity in the small intestine of rats fed non-purified and semipurified diets. AB - The effects of diet types on microsomal 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase in the intestinal villus and crypt cells isolated by scraping were studied in rats. The jejunoileal gradient for the reductase specific activity, a phenomenon usually observable in rats fed a commercial non-purified diet, disappeared when sucrose- or glucose-enriched semipurified diets were ingested. Feeding these semipurified diets resulted in a significant increase in reductase activity in jejunal villi and crypts with a concomitant decrease in that of the ileal populations. Neutral detergent fiber prepared from the non purified diet, but not cellulose powder, when included to semipurified diets at the levels equivalent to those in the non-purified diet, eliminated the ileojejunal gradient, reductase activity was virtually the same in the proximal and distal portions of the intestine or the distribution pattern resembled that shown in rats fed a non-purified diet. Potato and corn starches replaced for sucrose in a low-fiber semipurified diet were also effective for elimination of the gradient, but wheat starch was less effective in the respect. It is clear that intestinal sterogenesis via HMG-CoA reductase is specifically regulated by the complex interplay of dietary components, in particular carbohydrate sources. PMID- 6275056 TI - Composition and protein quality of Lupinus mutabilis. AB - The chemical composition and the protein quality of three samples of Lupinus mutabilis (a raw, semi-sweet variety; cooked, water-extracted seeds; and alcohol extracted oil cake) were studied. The protein content varied from 47.7% dry weight (raw seeds) to 65.3% (oil-cake). Compared to the FAO reference pattern sulfur- containing amino acids are first limiting. The water-extracted sample contained 26.9% oil and the polyunsaturated/saturated fatty acid ratio of 30 seed samples was 5.3. Alkaloid content of raw seed was high (3.3%), but could be reduced or nearly eliminated by water-and-alcohol extraction or plant breeding. Other anti-nutritive substances were present only in trace quantities. Protein quality measured as protein efficiency ratio (PER) gave low values for the non supplemented lupin proteins (1.34 semi-sweet variety; 1.53 water-extracted seeds; 1.19 oil-cake; 3.09 casein), but the PER's were improved by the addition of 0.2% DL-methionine to the diets (3.05, 2.69, 2,81, respectively). Raw as well as processed lupin protein showed an excellent apparent digestibility (80.0-85.8%, casein 87.1%). Studies of net protein utilization (NPU) and biological value (BV) confirmed the importance of methionine supplementation, The true digestibility of 92% was equivalent to that of casein. PMID- 6275057 TI - Survey of industrial workers for antibodies to toluene diisocyanate. PMID- 6275058 TI - Causes of death among employees of a synthetic abrasive product manufacturing company. AB - Analysis of the causes of death among workers employed in the manufacture of synthetic abrasive products was performed using data from death certificates and employment records. The subjects were 1,030 white male workers who had been employed by a single manufacturer and who were identified through a review of death certificates issued during a 20-year period. Proportional mortality analysis revealed excess digestive cancer and respiratory disease deaths. Personnel records permitted refinement of the study population to 968 with classification of each individual into one of seven employment categories. Case control analysis of workers ever employed in the individual categories revealed elevated odds ratios for respiratory disease deaths among those most exposed to the synthetic abrasive dust. The excesses were greatest in those exposed 20 or more years. Excesses of esophageal and rectal cancers were noted in two manufacturing areas although the numbers were small. Review of the literature further supports the findings that synthetic abrasive exposures may be associated with elevated respiratory morbidity and mortality. PMID- 6275059 TI - Amyloid in a pleomorphic adenoma of the parotid gland. Electron microscopic observations. PMID- 6275060 TI - Hepatic collagen synthesis in a rat model of cirrhosis, and its modification by colchicine. AB - Carbon tetrachloride inhalation in phenobarbitone treated rats caused a rapid rise in hepatic prolyl hydroxylase activity which was followed by an increase in hepatic collagen and free proline concentrations. Colchicine in a dose of 5 micrograms/day largely prevented the increase in hepatic collagen. This effect was not mediated by impairment of prolyl hydroxylase activity. Colchicine is of potential therapeutic value in preventing the progression of chronic liver disease to cirrhosis. PMID- 6275061 TI - Curved striated and round bodies in glomerulopathies of children; an ultrastructural study. AB - Extracellular curved striated bodies and round bodies were found in 62 and 41 per cent, respectively of 360 renal biopsies performed in children with a variety of nephropathies. They were observed in the basal lamina and mesangial matrix. The fact that they commonly occurred together appears to suggest a common aetiology and they are probably derived from altered protein of the basal lamina and mesangial matrix, or from proteins of immune deposits. PMID- 6275062 TI - Collagenase and neutral metallo-proteinase activity in extracts of inflamed human gingiva. PMID- 6275063 TI - A highly sensitive and specific assay for vertebrate collagenase. PMID- 6275064 TI - Calcium and the alpha-action of catecholamines on guinea-pig taenia caeci. AB - 1. The involvement of calcium in the alpha-action of adrenaline on guinea-pig taenia caeci was studied by measuring the changes in membrane potential and muscle contraction, using the sucrose-gap method, and by determining the (42)K efflux, in the presence of a beta-blocker (propranolol, 1.8 x 10(-6)m).2. In the presence of extracellular calcium, the hyperpolarization caused by adrenaline (3 x 10(-6)m) was sustained during the period of its application (5 min), both in active preparations (at 36 degrees C) and in quiescent muscle (22 degrees C). In the absence of calcium, adrenaline caused a transient hyperpolarization which was smaller at 36 degrees C than at 22 degrees C and passed off within 5 min, while adrenaline was present.3. Both the sustained and the transient hyperpolarization were associated with an increase in (42)K efflux which had a similar time course. (42)K flux measurements were made in depolarized tissue (52.8 mm-potassium), in which the effect was consistent and more pronounced than in polarized muscle (2.8 mm-potassium).4. The transient hyperpolarization which is resistant to calcium removal and EGTA (0.1-2.0 mm) could be evoked only once but, following a short exposure to calcium (2.5 mm) for 20 sec and readmission of calcium-free medium, it was restored.5. The sustained and the transient hyperpolarization and the increase in (42)K efflux were abolished by the alpha-antagonist phentolamine (10( 5)m); their amplitude was dependent on the adrenaline concentration in the range 10(-7) to 3 x 10(-6)m, and both responses persisted in the absence of sodium or chloride.6. The hyperpolarization and the increase in (42)K efflux were greater at higher external calcium concentrations (0.3-2.5 mm).7. Cobalt (0.6 mm), D600 (2.5 x 10(-5)m) and the bee toxin apamin (10(-7)m) reduced the alpha-response.8. In the presence of apamin, in calcium-containing solution, the sustained hyperpolarization caused by adrenaline was preceded by, or converted to, depolarization, spike discharge and contraction.9. The depolarizing effect of adrenaline in the presence of apamin persisted in sodium-free or chloride-free medium, but was blocked in the absence of calcium and diminished by cobalt and D600.10. It is concluded that the alpha-response of guinea-pig taenia caeci consists of two components, both involving calcium. First, the activation of alpha-receptors increases calcium entry, which leads to the opening of potassium channels, a sustained hyperpolarization and inhibition of muscle activity. Secondly, in the absence of external calcium, a transient hyperpolarization is revealed, presumably due to the release of bound calcium from a limited cellular store which can be replenished by addition of external calcium, and this leads to an increase in potassium permeability. PMID- 6275065 TI - A study of desensitization of acetylcholine receptors using nerve-released transmitter in the frog. AB - 1. Desensitization of acetylcholine (ACh) receptors was studied at the frog neuromuscular junction under voltage clamp.2. ACh was applied directly to junctional receptors by stimulating the motor nerve with trains of impulses. End plate currents (e.p.c.s) were used to estimate the total number of channel openings by the junctional ACh receptors, and miniature end-plate currents (m.e.p.c.s) were used to measure changes in post-synaptic sensitivity. Under the conditions of these experiments the changes in m.e.p.c. amplitudes were shown to be post-synaptic in origin and thus provided a measure of desensitization.3. When the acetylcholinesterase was inhibited with diisopropylfluorophosphate, neostigmine, or collagenase treatment to prolong the duration of the nerve released ACh in the synaptic cleft, desensitization developed during repetitive stimulation of 1000 impulses at 5-33 impulses/sec and then recovered after the conditioning trains, with a time constant of about 25 sec.4. When the acetylcholinesterase was active so that the duration of ACh in the synaptic cleft resulting from each nerve impulse was brief (< 300 musec), desensitization developed in response to 300-500 pairs of nerve stimuli if the interval between the impulses of each pair was 25 msec or less. When the interval was 30 msec or greater, however, measurable desensitization did not occur, even if the total number of channel openings was many times greater than in the experiments with shorter intervals or inhibited esterase where desensitization readily occurred.5. The desensitization observed to pairs of impulses was enhanced by chlorpromazine and decreased when the post-synaptic membrane was depolarized, properties similar to those described previously for desensitization to bath and ionophoretic application of ACh.6. These results indicate that desensitization to nerve released transmitter is not a simple consequence of receptor activation, is not due to blockade of the open receptor channels by ACh, and does not result from ACh binding directly to desensitized receptors with a resulting shift in the receptor population towards the desensitized state.7. We suggest that the desensitization observed to nerve-released transmitter is a two-step process with both steps initiated by ACh. In the first step ACh converts some receptors into a desensitizable state which has an apparent lifetime of less than 30 msec; in the second step ACh desensitizes the desensitizable state. PMID- 6275066 TI - Mechanistic implications of the potassium-potassium exchange carried out by the sodium-potassium pump. AB - 1. The magnitude of the K-K exchange carried out by the Na pump in human red cells was measured as a function of the external K concentration in cells with high and low intracellular K. The apparent K1/2 for external K and the Vm increased by the same proportion when intracellular K was raised. The result suggests that the K-K exchange is part of a ping-pong reaction mechanism. 2. The velocity of the exchange increases monotonically with intracellular phosphate and ATP concentration; neither substrate inhibits at very high concentration. 3. These results seem to require that the enzyme species which carries out the exchange is both phosphorylated and combined with ATP. PMID- 6275067 TI - Simultaneous changes in the equilibrium potential and potassium conductance in voltage clamped Ranvier node in the frog. AB - 1. In voltage clamped myelinated nerve fibres, the K+ conductance has been calculated from current recordings obtained in low and high K+ media, taking into account the changes in EK resulting from accumulation of depletion of K+ ions near to nodal membrane. 2. At the end of a depolarization, the instantaneous K+ current reverses at a potential (instantaneous reversal potential) differing from the Nernst potential calculated using the external and internal bulk concentrations (theoretical Nernst potential). During a depolarization, EK, as estimated from the instantaneous reversal potential, changes continuously. This change depends on the size, the duration and the direction of the time dependent K+ current. The variation of EK is attributed to continuous changes in K+ concentration near the membrane during voltage pulses which turn on the K+ conductance. 3. The chord conductance [GK = IK/(E-EK), as calculated using the instantaneous reversal potential values for EK, has been analysed as a function of time and membrane potential. As previously reported it increases with the initial K+ concentration in the external medium. 4. The time course of the K+ current depends on both the kinetics of the conductance increase and the rate of change in the driving force for K. The kinetics of the conductance increase can satisfactorily be described by a single exponential function following a delay after the onset of the depolarizing voltage clamp pulse. 5. This delay increases when the holding potential is made more negative. It decreases with membrane depolarization and it is independent of the external K+ concentration. At a given membrane potential, the turning on of the K+ conductance is found to be faster at high than at low external K+ concentrations. 6. At repolarization the turning off of the conductance cannot be described by a single exponential function. It is faster at low than at high external K+ concentrations. 7. The results suggest that the change in K+ conductance proceeds in a multi-step transition or (and) that the K+ conductance is determined by several types of K+ channels. PMID- 6275068 TI - Evidence for the existence of three types of potassium channels in the frog Ranvier node membrane. AB - 1. In voltage clamped myelinated fibres, the K+ current was recorded in high-K+ media to allow analysis without complications due to K+ accumulation. 2. After a depolarization, the tail of K+ current following repolarization decreases in two phases: a fast phase lasting about 20 msec and a slow exponential phase lasting several hundred milliseconds. When the duration of the depolarization is increased, the amplitude at time zero of the fast phase increases (activation of the conductance) and then decreases slowly (inactivation of the conductance). Simultaneously, the amplitude of the slow phase, extrapolated to time zero of repolarization, increases slowly and reaches a steady-state level (about 20% of the maximum instantaneous current) after about 600 msec of depolarization. 3. The fast phase of the tail current is blocked by external application of 4 aminopyridine (4-AP) (KD = 10(-5)M). The slow phase is unaltered by 4-AP (10(-7) 10(-2)M). 4. In the presence of 4-AP (10-3M), the remaining slow K+ current, activated by depolarizations, does not inactivate. 5. During depolarizations and repolarizations, the conductance of the slow current (GKs) varies exponentially. The steady-state value of the slow conductance and its time constant of activation vary with voltage. The variation of the slow conductance with time and voltage can be described by a closed-open mode, assuming that each channel is gated by one particle. The activation kinetics of the slow current is unaltered by long lasting (500 msec) prepolarizations. 6. The fast K+ conductance, calculated from the fast tail current, is fully inactivated at the end of a 3 min depolarization to 0 mV. 7. The fast K+ conductance can be decomposed into two components: one component (GKf1) activating between -80 and -30 mV and inactivating very slowly (tau = 45 sec at E = 0 mV); one component (GKf2) activating between -40 mV and +30 mV and inactivating slowly (tau = 2 sec at E = 0 mV). t = 12 degrees C. 8. The maximum slow and fast conductances increase with [K]0. While the maximum fast conductance tends to saturate at high external K+ concentrations, the maximum slow conductance shows no sign of saturation. 9. A comparison between motor and sensory fibres shows that, while the amplitude of maximum slow and fast conductances are identical for both types of fibres, the amplitude of fast-1 conductance is larger and consequently the amplitude of fast 2 is smaller in motor than in sensory fibres. The different spike frequency adaptations observed on both types of fibres are discussed in relation to these different relative fast conductances amplitudes. 10. It is concluded that the K+ conductance of the nodal membrane is composed of three components (GKS, GKf1 and GKf2) corresponding to three different and distinct types of K+ channels. PMID- 6275069 TI - Restoration of focal multiple innervation in rat muscles by transmission block during a critical stage of development. AB - 1. The soleus, extensor digitorum longus and peroneus tertius muscles of the hind leg were paralysed by botulinum toxin injection in neonatal rats varying in age from 8 to 31 days. The soleus muscle was similarly paralysed in adult rats.2. Muscles were excised after different periods of block, neuromuscular transmission was assessed in vitro, and the nerve terminal size and amount of terminal sprouting and multiple innervation determined histologically using silver (Ag) and zinc iodide-osmium (ZIO) tetroxide stains.3. Recovery from the block induced by botulinum toxin was more rapid in immature multiply-innervated muscles than in muscles of older rats in which all multiple innervation had been eliminated.4. Terminals in muscles blocked during the first month after birth grew rapidly in size and developed a characteristic granulose morphology in the first few days following the block. This change did not occur in fully adult rat solei.5. Sprouts growing from the terminals were infrequent in muscles paralysed before day 16. Terminal sprouts were more frequent in muscles paralysed between 16 and 31 days of age, but were very infrequent in adult solei.6. In confirmation of Thompson, Kuffler & Jansen (1979) paralysis begun at 10 days was followed initially by a continued fall in multiple innervation detected electrophysiologically. After 2 days the percentage of muscle fibres with more than one input rose. The extra inputs did not come from terminal sprouts. They innervated the single synaptic site on each muscle fibre as they do during normal synaptogenesis.7. The amount of multiple innervation regained 5-10 days after the start of paralysis became progressively less the later paralysis started and was approximately equal to the level existing at the time the block was begun. Thus paralysis starting on or after day 16 (when all excess inputs have normally withdrawn) caused no return of multiple innervation.8. The return of multiple inputs and the swelling of the nerve terminals are presumably responses of the motoneurone to growth stimuli from inactive muscle. It is not clear whether the return of multiple innervation in the 10-15 day old rats is due to reactivation of inputs still in close contact with the end-plate before withdrawal or to regrowth of partly retracted nerve branches.9. A parallel is drawn between the limited period when inactivity can reinstate multiple innervation, and the critical period in the developing visual cortex. PMID- 6275070 TI - The effect of burst patterning of preganglionic input on the efficacy of transmission at the cat stellate ganglion. AB - 1. The effect of burst patterned preganglionic stimulation on the efficacy of transmission at the cat stellate ganglion in vivo was studied to determine whether the increase in acetylcholine (ACh) release that occurs at preganglionic terminals with similar stimulation is synaptically active.2. Stimulation of the whole or a portion of the thoracic sympathetic trunk between rami T4 and T5 with recording of evoked compound action potentials in the inferior cardiac nerve yielded a preparation with a subliminal fringe with a mean value 2.5 times the size of the discharge zone.3. Preganglionic stimulation with 0.5 s bursts at 10, 20 and 40 Hz with 10 s interburst intervals caused an increase with time in the area of the compound action potential evoked by a single interburst test stimulus. The increase reached 70-75% of the final value after the first burst, 75-90% after 5 bursts and was virtually complete after 5 min. It was then maintained without attenuation as stimulation was continued.4. On cessation of burst patterned stimulation test compound action potentials returned to control levels with a half-time of 5 min.5. The increase in area of test compound action potentials with 40 Hz intraburst frequency was always as great as the increase following tetanization of the pathway. The increases with 20 and 10 Hz bursts were less.6. There was no increase in area of compound action potentials when equally spaced pulses at 2 Hz or less were applied preganglionically, nor was there any increase when 40 Hz burst patterns were delivered directly to the inferior cardiac nerve.7. It is concluded that burst patterned preganglionic activity at frequencies similar to those observed in vivo can recruit rapidly into the discharge zone all of the ganglionic neurones that were originally in the subliminal fringe. It is proposed that this effect is related directly to the increase in ACh release at ganglionic synapses that such activity has been shown to induce. PMID- 6275072 TI - A further study of the phospholipase-independent action of beta-bungarotoxin at frog end-plates. AB - 1. The effect of beta-bungarotoxin (beta-BuTx) at the frog neuromuscular junction has been investigated further in order to distinguish more clearly between phospholipase- independent and phospholipase-dependent actions on transmitter release. 2. Inhibition of the enzymatic activity, by substitution of strontium for calcium, allowed determination of the dose-response curve of the early rapid decrease in transmitter release caused by the toxin. In the presence of strontium ions there was, however, still about 7% residual enzymatic activity, and electrophysiological evidence of it could be seen in room-temperature experiments at high concentrations of beta-BuTx. This residual enzymatic activity could be suppressed by lowering the temperature to 5 degrees C. 3. In normal calcium Ringer solution beta-BuTx produced the typical triphasic effect on the amplitude of end-plate potentials (e.p.p.s). Lowering the temperature markedly delayed an then diminished the secondary transient increase. There was, however, comparatively little temperature influence on the first rapid decrease in e.p.p. amplitude. Enzymatic assays confirmed the temperature dependence of the toxin's phospholipase activity on model phospholipid substrates. 4. The kinetics of the phospholipase-independent action of beta-BuTx were examined in strontium-Ringer compared to calcium-Ringer solution, as well as in calcium-Ringer at different temperatures. Both the time to onset of inhibition and the time to 50% inhibition of the e.p.p., during the first phase of toxin action, are temperature-dependent and briefer in calcium than in strontium-Ringer solution. It is suggested that calcium is more effective than strontium in promoting this phospholipase- independent interaction of beta-BuTx with the nerve terminal membrane. PMID- 6275071 TI - Multiple receptor types for octopamine in the locust. AB - 1. Three different pharmacological classes of octopamine receptor mediate the actions of octopamine on the locust extensor-tibiae neuromuscular preparation. A receptor classification scheme is proposed based on the results of detailed studies with agonists and antagonists. 2. Octopamine1 class receptors mediate the slowing of a myogenic rhythm found in a specialized proximal bundle of muscle fibres. Octopamine2A class receptors mediate the increase in amplitude of slow motoneurone twitch tension and octopamine2B class receptors mediate the increase in relaxation rate of twitch tension induced by firing either the fast or the slow motoneurones. 3. Octopamine1 receptors can be distinguished from the 2A and 2B classes since chlorpromazine (and yohimbine) are much better blocking agents than metoclopramide at the former receptors, whereas the converse is true for the latter class. Also clonidine is a more effective agonist than naphazoline for the former receptors and the converse is true for the latter class. 4. Octopamine 2A can be distinguished for octopamine 2B receptors since metoclopramide, mianserin and cyproheptadine show a strong preference for blocking the former class. Also naphazoline is a much better agonist than tolazoline at the former receptors and tolazoline is a much better agonist than clonidine at a latter. 5. The results are discussed in terms of the location of the various classes of octopamine receptors, their possible relationship to vertebrate alpha-adrenoreceptors, and the significance of the results for studies on octopamine receptors in the vertebrate central nervous system. PMID- 6275073 TI - beta-Receptor numbers and thermodynamics in denervation supersensitivity. AB - 1. Denervation supersensitivity to adrenergic agonists occurs after degeneration of the sympathetic nervous system in the disease called multiple system atrophy (MSA) or the Shy Drager Syndrome.2. Supersensitivity to the chronotropic effect of i.v. isoprenaline on the heart was demonstrated in eight subjects with sympathetic nervous system degeneration and MSA.3. There was an increased number of beta-receptors present in MSA as measured by [H(3)]dihydroalprenolol ([H(3)]DHA) binding to beta-receptors on lymphocytes isolated from venous blood taken from the MSA subjects compared with [H(3)]DHA binding to lymphocytes from seven normal subjects. There was no difference in the affinity of lymphocyte beta receptors for [H(3)]DHA in MSA.4. [H(3)]DHA binding to lymphocytes from MSA subjects was decreased at lower temperatures but was unaffected by lower temperatures in lymphocytes from normal subjects.5. Equilibrium constants for [H(3)]DHA binding to normal and MSA lymphocytes were similar, indicating that the affinity of the beta-receptors was similar in both groups. The equilibrium constants were little affected by cooling from 37 to 4 degrees C suggesting that the heat of reaction (enthalpy) for [H(3)]DHA binding was low. The Gibbs free energy change on binding was negative and similar in quantity for both normal and MSA lymphocytes. There was a similar, large, increase in entropy on binding of [H(3)]DHA to both normal and MSA lymphocytes, showing that the binding reaction was entropy driven.6. If lymphocyte beta-receptors reflect the status of cardiac beta-receptors, increased numbers of cardiac beta-receptors may contribute to the denervation supersensitivity to isoprenaline in MSA with sympathetic degeneration. PMID- 6275074 TI - Actions of indomethacin and prostaglandins on neuro-effector transmission in the dog trachea. AB - Neuro-effector transmission in the smooth muscle layer of the dog trachea was studied in vitro using the micro-electrode and double sucrose gap methods.1. Electrical field stimulations with short duration (50-100 musec) applied to the whole tissue produced an excitation of the intrinsic nerves, and evoked excitatory junction potentials (e.j.p.s) followed by twitch tension development and subsequent long lasting relaxation of the smooth muscle tissue.2. The effects of field stimulations were abolished by tetrodotoxin (2 x 10(-7)m), and atropine (1.7 x 10(-5)m) selectively blocked both the e.j.p. and twitch tension. On the other hand, propranolol (1.9 x 10(-5)m) suppressed the generation of the prolonged relaxation evoked by the field stimulations.3. E.j.p.s recorded by the double sucrose gap method showed gradual and continuous reduction in amplitude during prolonged exposure in Krebs solution (1-2 hr), and there were no changes in the membrane potential or in the input membrane resistance.4. With application of indomethacin (10(-5)m), a gradual and continuous reduction in the amplitude of e.j.p. was no longer observed, and (after the initial increase in the amplitude) e.j.p.s with a constant amplitude were obtained during 1-1.5 hr. Indomethacin (10(-5)m) modified neither the resting membrane potential nor the input membrane resistance of smooth muscle cells.5. After pre-treatment with indomethacin, low concentrations (10(-11)-10(-8)m) of prostaglandin E(1) or E(2) (PGE series) markedly suppressed the amplitude of e.j.p. with no changes in the resting membrane potential or in the input membrane resistance.6. During the repetitive field stimulation at the stimulus frequency of 0.1-1 Hz, the amplitude of the e.j.p.s was gradually reduced (the depression process). The depression was not affected by applications of prostaglandins, indomethacin or alpha- and beta adrenoceptor blockers.7. These results indicate that in the dog tracheal smooth muscles, the endogenous PGE series may play an important role in feed-back inhibitory mechanisms, at the nerve terminals related to acetylcholine release. PMID- 6275075 TI - Calcium current-dependent and voltage-dependent inactivation of calcium channels in Helix aspersa. AB - 1. Inactivation of the Ca channels has been examined in isolated nerve cell bodies of Helix aspersa using the suction pipette method for voltage clamp and internal perfusion.2. Satisfactory suppression of outward currents was essential. This was achieved over most of the voltage range by substitution of Cs ion for K ion and by the use of TEA intra- and extracellularly and 4-AP extracellularly. A small time- and voltage-dependent non-specific current remained at potentials above +60 mV.3. In these solutions, Ca current approaches E(Ca) but cannot be detected in the outward direction. The Ca channel appears to be impermeable to Cs and Tris ions.4. Inactivation of Ca currents occurs as a bi-exponential process. The faster rate is 10-20 times the slower rate and is about one twentieth the rate of activation. The development of inactivation during a single voltage-clamp step and the onset of inactivation produced by prepulses followed after brief intervals by a test pulse, have roughly similar time courses.5. The rates of inactivation increase monotonically at potentials more positive than about -25 mV. The amount of steady-state inactivation increases with membrane depolarizations to potentials of about +50 mV. At more positive potentials, steady-state inactivation is reduced.6. Intracellular EGTA slows the faster rate of inactivation of I(Ca) and reduces the amount of steady-state inactivation measured with a standard two pulse protocol. The effect is specifically related to Ca chelation and hydrogen ions are not involved. This component of inactivation is referred to as Ca current-dependent inactivation and is consistent with observations that increased Ca(i) inactivates the Ca channel. The process does not depend upon current flow alone since Ba currents of comparable or greater magnitude have smaller initial rates of inactivation. Furthermore, application of Ba ion intracellularly in large concentrations has no effect on steady-state inactivation.7. The bi-exponential inactivation process that persists in the presence of EGTA(i) is similar to that occurring when extracellular Ba ion carries current through the Ca channel. Steady-state inactivation also persists and is similar in the two cases. Therefore it is concluded that inactivation is voltage-dependent as well as Ca current dependent.8. Diffusion models that included reasonable values for the effect of binding on diffusion, even when combined with declining influxes, did not account for this ;mixed' form of calcium- and voltage-dependent inactivation. A compartmental model in which the particular kinetic model of voltage-dependent inactivation was not critical described the Ca current-dependent inactivation. PMID- 6275076 TI - Parameters affecting the slow inward channel repriming process in frog atrium. AB - 1. The time of recovery (from the inactivation) of the slow inward current was studied in the frog atrium, using the double sucrose gap voltage clamp technique. 2. The 'repriming' process was found to be distinct from the current inactivation, and to depend on experimental protocol: double pulses given at low frequencies (at 'rest') gave a faster recovery time when compared to recovery during constant stimulation, with interposed stimuli monitoring the recovery. Longer durations of the clamp pulses led to a faster recovery process. 3. Changing the holding potential of the membrane (with double pulses to the same absolute membrane potential monitoring the recovery process) greatly affect the repriming with depolarized levels slowing down the process. 4. The recovery time was fastest following clamp pulses to intermediate membrane potentials (in the plateau range). This was determined by double pulses, from a constant hold potentials, to different levels. 5. Decreasing extracellular Ca prolonged, and increasing Ca enhanced the recovery process. 6. The recovery process was markedly slowed down in Na or in K-free solutions. 7. The recovery process was enhanced in solutions with a raised concentration of Mg or H ions (lower pH). In higher Mg solutions, the inactivation of the slow inward current was slower. 8. It is proposed that the recovery process is sensitive to alterations in intracellular Ca ions and to variations in extracellular surface charges. The possible implications are discussed. PMID- 6275077 TI - Evidence that histamine and carbachol may open the same ion channels in longitudinal smooth muscle of guinea-pig ileum. AB - 1. Membrane potential was recorded intracellularly by micro-electrode in separated longitudinal muscle of guinea-pig ileum. Electrotonic potentials were evoked in longitudinal strips by passing current between large external electrodes in the partition chamber.2. Histamine increased the frequency of action potential discharge at low concentrations and depolarized the membrane. At higher concentrations it caused substantial depolarization and action potential discharge was abolished. Carbachol had similar actions but the maximal depolarization by carbachol (using 10(-4)m) was some 4-5 mV greater than maximal depolarization by histamine (using 10(-4)m).3. The change in size of evoked electrotonic potentials was used to estimate the effects of carbachol and histamine on the conductance of the smooth muscle membrane. The equilibrium potentials for histamine and carbachol depolarizations were estimated from their relative effects on potential and conductance and were found to be not significantly different; measurements of the effects on conductance showed that 10(-4)m-histamine increased conductance about 8-fold whilst 10(-4)m-carbachol had a much greater effect on conductance. This difference could explain the differing maximal depolarizing effects of these agents if both were assumed to open channels having the same ionic selectivity (i.e. equilibrium potential).4. The efflux of (42)K was studied in separated strips of longitudinal ileal muscle from guinea-pig. In the presence of a concentration of carbachol (2 x 10(-5)m or 10( 4)m) having a maximal effect on (42)K efflux rate, histamine (10(-4)m) did not increase efflux further although 120 mm-potassium did so. Experiments with the irreversible muscarinic receptor blocker, propylbenzilylcholine mustard, indicated that the number of muscarinic receptors did not limit the (42)K efflux response to carbachol and it was suggested that the response was limited by the availability of ion channels which could be opened by activated muscarinic receptors.5. Contractions to histamine and carbachol in 120 mm-potassium depolarizing solution were followed upon washing by a relaxation below basal tension. Carbachol, but not histamine, showed a pronounced and long lasting secondary contraction following this relaxation.6. These results are consistent with the idea that activated histamine and activated muscarinic receptors open the same ion channels in the smooth muscle membrane to produce depolarization, increased action potential discharge and contraction, although muscarinic receptors can open more of these. However, there was evidence that the opening of these channels is not the only pathway between receptor activation and contraction. PMID- 6275078 TI - Action of 5-hydroxytryptamine on intestinal ion transport in the rat. AB - 1. 5-HT increased the electrical activity of rat jejunum both in vivo and in vitro. The increased potential difference and short-circuit current resulted from a stimulation of electrogenic chloride secretion. NaCl absorption may also have been inhibited. 2. 5-HT did not alter cyclic AMP levels in isolated enterocytes. 3. The 5-HT response in vivo was unaffected by atropine, cyproheptadine, propranolol and hexamethonium. Phenoxybenzamine reduced the maximum response without affecting the dose required to produce a 50% maximum response. Methysergide, at a dose of 40 mg/kg, had a similar effect while a lower dose of 2 mg/kg produced no change. Mianserin competitively antagonized the response to 5 HT, a dose of 2 mg/kg producing a fourfold increase in the amount of 5-HT required to produce a 50% maximum response. 4. Acetylcholine and 5-HT seem to act independently in inducing intestinal secretion since atropine did not block the response to 5-HT and Mianserin did not alter the response to acetylcholine. 5. Experiments in which the intestinal villi or crypts were subjected to preferential damage suggested that 5-HT primarily produced its response at the crypt cell level. PMID- 6275080 TI - Chemotherapy of influenza and herpes virus infections. PMID- 6275079 TI - Effect of Escherichia coli heat-stable enterotoxin, cholera toxin and theophylline on ion transport in porcine colon. AB - 1. The effect of heat-stable enterotoxin (ST) of Escherichia coli, cholera toxin (CT), and theophylline (a phosphodiesterase inhibitor) on ion and water transport was studied with an in vivo isolated loop system of the pig colon.2. All three agents abolished net Na absorption as a result of a decrease in the lumen to blood Na flux alone. With all three agents, net Cl absorption was reduced, but not abolished, and net HCO(3) secretion was elicited. Luminal p(CO2) was reduced with CT and theophylline from that observed in normal Ringer alone.3. Theophylline resulted in a prompt and sustained increase in both cyclic adenosine monophosphate (cyclic AMP) and cyclic guanosine monophosphate (cyclic GMP) levels in colonic mucosa studied in vitro. ST selectively elevated cyclic GMP, whereas CT selectively elevated cyclic AMP. These responses paralleled the time course and magnitude of response of the transepithelial electrical potential difference (psi(LB)) measured in vivo.4. Ion replacement studies in the presence or absence of theophylline showed that in the absence of Na, Cl absorption was slightly reduced and HCO(3) secretion was elicited; no further additive effects of theophylline in the absence of luminal Na were observed. In the absence of luminal Cl, net Na absorption was abolished and HCO(3) was absorbed; theophylline resulted in significant net Na and HCO(3) secretion. Theophylline also increased psi(LB) in the absence of either luminal Na or Cl.5. Results suggest that in the presence of theophylline or enterotoxin, the coupled Na-H and Cl-HCO(3) exchange processes that are normally responsible for at least half of the net NaCl absorption by this tissue are interrupted. Active HCO(3) secretion is observed and Cl absorption under these conditions can be entirely explained as a consequence of psi(LB). Thus, these studies indicate that the colon may participate in the production of diarrhoea of enterotoxigenic origin. They also suggest an important functional role of cyclic nucleotides in controlling the acidity and volume of colonic contents. PMID- 6275081 TI - [Computed tomography finding in circumscribed lesions of the liver (author's transl)]. PMID- 6275082 TI - Response of the epididymal duct in the corpus epididymidis to efferent or epididymal duct ligation in the mouse. AB - Different parts of the epididymal duct were ligated when mice were 90 days old. The mice were killed 1--4 weeks later. PAS-positive materials appeared in the epithelial cells of Segment IV (corpus epididymidis) after ligation of the efferent ducts or at Segment II (middle part of caput) but not when the ligature was distal to Segment II. The inclusions were seen as early as 1 week after ligation and became increased in size and number with time. PMID- 6275083 TI - Proteolytic enzymes and gonadal hormones of the ovarian follicle wall during ovulation in the domestic fowl (Gallus domesticus). AB - There were no significant changes in the activities of collagenase, acid and neutral protease up to ovulation. Neutral protease activity increased significantly in the post-ovulatory follicle obtained immediately after ovulation. Acid protease and collagenase showed an increasing activity with time after ovulation. The progesterone concentration of the follicle wall rose 3 h before ovulation, peaked 1 h before ovulation and remained high until ovulation. Plasma progesterone values were high but decreased before those in the follicle. Follicular progesterone concentrations decreased markedly soon after rupture, but oestradiol values then increased. PMID- 6275084 TI - Inhibition of ovulation and LH secretion in the gilt after treatment with ACTH or hydrocortisone. AB - Treatment with ACTH (100 i.u.) or hydrocortisone acetate (250 mg) twice daily for 12 days to increase cortisol concentrations blocked ovulation in all gilts. The preovulatory surge of LH was also blocked in the treated gilts. Oestrous cycle length and number of days in oestrus were similar for all treatments except that in one of the 2 experiments ACTH suppressed oestrus in 4 of the 5 gilts treated. PMID- 6275085 TI - Silica stimulation of chemotactic factor release by guinea pig alveolar macrophages. PMID- 6275086 TI - Isolation of Chinese hamster cell mutants defective in the receptor-mediated endocytosis of low density lipoprotein. PMID- 6275087 TI - Binding of RNA polymerase and the catabolite gene activator protein within the cat promoter in Escherichia coli. PMID- 6275088 TI - Evidence that cyclic GMP may regulate cyclic AMP metabolism in the isolated frog ventricle. PMID- 6275089 TI - Mitochondrial acyl-CoA, adenine nucleotide translocase activity and oxidative phosphorylation in myocardial ischaemia. PMID- 6275090 TI - Simian virus 40 as a probe for studying inducible repair functions in mammalian cells. AB - We describe the use of Simian Virus 40 (SV40) as a molecular probe for studying the cellular functions induced in cultured monkey kidney cells in response to DNA damaging agents. (a) Ultraviolet (UV) irradiation of SV40-infected cells inhibits viral DNA replication. Replication forks are blocked by the first pyrimidine dimer encountered. In some cases, a single-strand break seems to occur at the level of the dimer inhibiting the fork of replication. This break, which can be visualized by electron microscopy studies, might be the first step in an excision repair pathway. (b) Treatment of monkey kidney cells with acetoxy-acetyl aminofluorene or UV light before infection with UV-irradiated SV40 induces a mutagenic replication mode, as shown by an increase of the mutation frequency of thermosensitive SV40 mutants. (c) A possible recombination assay using various SV40 mutants infecting the same cell is proposed and discussed. PMID- 6275091 TI - On the presence of met 5-enkephalin receptors on the plasma membrane of Deiters' neurons and their modulation of GABA transport. AB - Met 5-enkephalin blocks the active transport of GABA across plasma membranes of rabbit Deiters' neurons prepared by microdissection. Such an effect is receptor mediated, being reversed by the antagonist Naloxone. This result shows that Deiters' neurons bear enkephalin receptors, and suggests a hypothesis on the mechanism of enkephalin-mediated neuronal inhibition. PMID- 6275092 TI - Maternal diet, breast feeding and infants' growth. A field study in the Ivory Coast (West Africa). PMID- 6275093 TI - Viral protein synthesis in mouse hepatitis virus strain A59-infected cells: effect of tunicamycin. AB - We identified eight protein species in virions of mouse hepatitis virus strain A59. Based on their sizes, prosthetic groups, and locations in virions, these proteins were designated gp180/E2, gp90/E2, pp54/N, gp26.5/E1, gp25.5/E1, p24/E1, p22/X, and p14.5/Y. The positions of the last two proteins in virions are not known. Host protein synthesis in Sac(-) cells infected with mouse hepatitis virus strain A59 was inhibited, and the following novel proteins appeared: gp150, gp90, p54, gp26.5, gp25.5, p24, p22, and p14.5. Except for gp150, these polypeptides all co-electrophoresed with mouse hepatitis virus strain A59 structural proteins. In addition, all of these proteins could be immunoprecipitated with a convalescent mouse serum or a rabbit antiserum raised against purified disrupted virus. After a 15-min pulse of infected cells with radioactive amino acids at 7h postinfection, gp90 was not detected, whereas gp26.5 and gp25.5 were only labeled to a small extent. During a subsequent chase period gp150 was processed to gp90, whereas the radioactivity in gp26.5 and gp25.5 increased concomitantly with a reduction of label in p24. Tunicamycin, an antibiotic which inhibits the synthesis of glycopeptides bearing N glycosidically linked oligosaccharides, prevented the appearance of gp150 in mouse hepatitis virus strain A59-infected cells. Instead, a 110,000-dalton protein accumulated. In contrast, the syntheses of the smaller viral glycoproteins gp26.5 and gp25.5 were resistant to this drug, indicating that these glycosylations were of the O glycosidical type. Although the production of infectious virus in tunicamycin-treated cells was inhibited by more than 99%, release of noninfectious viral particles continued. An analysis of these particles revealed that they lacked the peplomeric glycoproteins gp90/E2 and gp180/E2. Obviously, although the surface projections were not essential for budding of virus particles from the cells, they were required for infectivity. PMID- 6275094 TI - Proteins Specified by bovine herpesvirus 1 (infectious bovine rhinotracheitis virus). AB - An electrophoretic analysis of radioactively labeled, purified, "empty" and DNA containing infectious bovine rhinotracheitis virions revealed the presence of 25 to 33 structural (virion) polypeptides. A total of 11 of these polypeptides could be labeled with [3H]glucosamine and were identified as glycoproteins. In addition to the 25 structural polypeptides, infectious bovine rhinotracheitis virus infected cells also contained at least 15 nonstructural (nonvirion) polypeptides that were not present in purified virions. Expression of the viral polypeptides in infected cells was controlled temporally. Thus, most viral polypeptides could be categorized as "alpha" (immediate early), "beta" (early), or "gamma" (late) on the basis of their order of appearance in infected cells and whether their syntheses were dependent upon prior viral protein or DNA synthesis. None of the glycoproteins belongs to the alpha class, although at least one (GVP11) was synthesized in the absence of viral DNA synthesis. Serum from a cow in which infectious bovine rhinotracheitis virus lesions were reactivated by dexamethasone precipitated both structural and nonstructural polypeptides. PMID- 6275095 TI - Deletion of a 9,000-base-pair segment of the vaccinia virus genome that encodes nonessential polypeptides. AB - Deletions contained within the genomes of unstable and stable variants of vaccinia virus (strain WR) were analyzed. Restriction endonuclease mapping and hybridization to specific 32P-labeled DNA probes indicated that more than 6 X 10(6) daltons of DNA were deleted from the variants. In each case, the deletion occurred on the left side of the genome and started very close to the junction of the inverted terminal repetition and unique sequence. Both variants also contained a new SstI side on the right side of the genome. Hybridization selection and cell-free translation experiments indicated that these variants lost the ability to synthesize at least eight early mRNA's mapping within the deleted region. Although the deleted DNA was not essential for replication of the WR strain of vaccinia virus under laboratory conditions of infection, it presumably has a defined role under other circumstances. This conclusion was based on the conservation within the Elstree strain of vaccinia, the Utrecht strain of rabbitpox, and the Brighton strain of cowpox virus of sequences homologous to the deleted DNA. Moreover, mRNA's that hybridized to the deleted vaccinia virus DNA segment and encoded similar size polypeptides were made in cells infected with rabbitpox and cowpox viruses. PMID- 6275096 TI - Integration of Rous sarcoma virus DNA into chicken embryo fibroblasts: no preferred proviral acceptor site in the DNA of clones of singly infected transformed chicken cells. AB - We analyzed retroviral integration into a host genome by using avian sarcoma virus infection of natural target cells under conditions where secondary integration via virus spread was inhibited. This was accomplished by using the noninfectious pol- env- alpha variant of the Bryan high-titer strain of Rous sarcoma virus. A total of 12 independent Bryan high-titer Rous sarcoma virus transformed chicken embryo fibroblast clones were obtained and mapped by using restriction endonucleases. Provirus-cell junction fragments were identified with appropriate hybridization probes. We found that expression of the viral genes could occur after proviral integration at many sites on the chicken genome and that there was no apparent preference for specific integration sites. PMID- 6275097 TI - Structural organization and biological activity of molecular clones of the integrated genome of a BALB/c mouse sarcoma virus. AB - BALB/c mouse sarcoma virus (BALB-MSV) is a spontaneously occurring transforming retrovirus of mouse origin. The integrated form of the viral genome was cloned from the DNA of a BALB-MSV-transformed nonproducer NRK cell line in the Charon 9 strain of bacteriophage lambda. In transfection assays, the 19-kilobase-pair (kbp) recombinant DNA clone transformed NIH/3T3 mouse cells with an efficiency of 3 X 10(4) focus-forming units per pmol. Such transformants possessed typical BALB MSV morphology and released BALB-MSV after helper virus superinfection. A 6.8-kbp DNA segment within the 19-kbp DNA possessed restriction enzyme sites identical to those of the linear BALB-MSV genome. Long terminal repeats of approximately 0.6 kbp were localized at either end of the viral genome by the presence of a repeated constellation of restriction sites and by hybridization of segments containing these sites with nick-translated Moloney murine leukemia virus long terminal repeat DNA. A continuous segment of at least 0.6 and no more than 0.9 kbp of helper virus-unrelated sequences was localized toward the 3' end of the viral genome in relation to viral RNA. A probe composed of these sequences detected six EcoRI-generated DNA bands in normal mouse cell DNA as well as a smaller number of bands in rat and human DNAs. These studies demonstrate that BALB-MSV, like previously characterized avian and mammalian transforming retroviruses, arose by recombination of a type C helper virus with a well conserved cellular gene. PMID- 6275098 TI - Vertebrate DNAs contain nucleotide sequences related to the transforming gene of avian myeloblastosis virus. AB - Avian myeloblastosis virus contains a continuous sequence of approximately 1,000 nucleotides which may represent a gene (amv) responsible for acute myeloblastic leukemia in chickens. This sequence appears to have been acquired from chicken DNA and to be substituted for the envelope gene in the viral genome. We used hybridization probes enriched for the amv sequences and conditions that facilitate annealing of partially homologous nucleotide sequences to show that cellular sequences related to amv are present in the genomes of all vertebrates ranging from amphibians to humans but were not detected in fish, sea urchins, or Escherichia coli. In contrast to the preceding findings, nontransforming endogenous proviral nucleotide sequences closely related to the remainder of the avian myeloblastosis virus genome and to the entire myeloblastosis-associated helper virus are present only in chicken DNA. The amv-related cellular sequences appear to be highly conserved during evolution and to be contained at only one or a few locations in the genome of vertebrates. Within closely related species, they appear to share common evolutionary genetic loci. These findings and similar ones obtained with other highly oncogenic retroviruses containing a transforming gene suggest a general mechanism for acquisition of viral oncogenic sequences and an essential role for these sequences in the normal cellular state. PMID- 6275099 TI - Epstein-Barr virus DNA. X. Direct repeat within the internal direct repeat of Epstein-Barr virus DNA. AB - The 3,360-base-pair internal direct repeat (IR) in Epstein-Barr virus DNA separates the short and long unique DNA domains. IR has a single BamHI site. The juncture between the short unique domain and IR has been mapped by restriction endonucleases and is less than 2,600 nucleotides before the BamHI site in IR. The junction between IR and the long unique domain has been sequenced and is approximately 650 nucleotides after the BamHI site in IR. Thus, relative to the start of IR at the juncture with the short unique domain, the last repeat is at least 90 base pairs short of being complete. There is homology between the 250 nucleotide fragments to the left and the right of the unique BamHI site in IR. A 35-base-pair sequence of the left fragment is directly repeated within the right fragment, once fully and once partially. The implications of these findings are discussed. PMID- 6275100 TI - Structure of varicella-zoster virus DNA. AB - Varicella-zoster virus (VZV) DNA was prepared from nucleocapsids and from enveloped virions of a laboratory strain (Ellen) and directly from the vesicle fluids of patients with zoster infections. VZV Ellen nucleocapsid DNA was cleaved with 11 different restriction endonucleases and electrophoresed in agarose gels. The restriction profiles of the nucleocapsid DNA were identical to those of the DNA recovered from purified virions, but differed from those of another VZV strain (KM). In vitro-labeled VZV K.M. DNA purified directly from vesicle fluid yielded a distinct restriction pattern which appeared to be unchanged after several tissue culture passages of the isolate from that fluid. Restriction endonuclease analysis (EcoRI or BglII) of VZV DNA revealed the presence of four cleavage fragments with a molar ratio of approximately 0.5. No individual fragments with molar ratios of 0.25 were noted. This observation suggests that the VZV genome may contain one invertible segment. Comparison of the electrophoretic migrations of VZV DNA fragments relative to those of DNAs of known size permitted calculation of the VZV genome size to be 72 X 10(6) to 80 X 10(6) daltons. These results were confirmed by electron microscopy which demonstrated a genome size of about 76 X 10(6) daltons for passaged and unpassaged VZV DNA. Electron microscopy also revealed that some of the DNA molecules recovered from nucleocapsids or directly from vesicle fluids were superhelical circles. PMID- 6275101 TI - Genomic arrangement of an adenovirus-simian virus 40 hybrid virus, Ad2+ND4del. AB - Ad2+ND4del is an adenovirus type 2-simian virus 40 hybrid virus nondefective for growth in human cells. The virus was first observed when stocks of Ad2+ND4, a hybrid isolated from primary monkey kidney cells, were propagated in human cells. This paper describes the DNA sequence at two sites of DNA recombination, the site of the left adenovirus type 2-simian virus 40 junction and the site of a deletion of internal simian virus 40 sequences. Since the deletion was observed when the virus was switched from monkey to human cells, an analysis of gene expression in the region of DNA rearrangement may prove useful for the elucidation of molecular events that accompany virus growth in different hosts. PMID- 6275102 TI - Avian acute leukemia virus OK10 has an 8.2-kilobase genome and modified glycoprotein gp 78. AB - We have analyzed the structure of OK10-BM virus, an avian acute leukemia virus produced by a bone marrow-derived cell line of macrophage origin, and compared it with that of OK10 AV, an associated virus originally present in the OK10 virus stock. The RNAs of OK10-BM virus and OK10 AV had the same mobility in agarose gels, corresponding to 8.0 to 8.5 kilobases, a size considerably larger than that of the transforming component (5 to 6 kb) of most other avian acute leukemia viruses. Fingerprint analysis showed a close relationship between OK10-BM virus and OK10 AV RNAs. The polypeptide compositions of OK10-BM and OK10 AV viruses were similar except for the envelope glycoproteins. In analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the large envelope glycoprotein of OK10-BM virus migrated at M(r) = 78,000 (gp78), whereas OK10 AV had the characteristic 85,000-dalton glycoprotein (gp85) of nondefective avian leukemia viruses. gp78 was weakly labeled with methionine, glycine, proline, or mannose, suggesting that purified OK10-BM virus had reduced amounts of the modified envelope glycoprotein. In cell-free rabbit reticulocyte lysates, OK10-BM virion RNA directed the synthesis of a 200,000-dalton polypeptide (p200), a 180,000 dalton polypeptide (pr180), and a 76,000-dalton polypeptide (pr76), whereas OK10 AV RNA gave rise only to pr180 and pr76, suggesting that p200 may represent an OK10-BM-encoded transforming protein. No biochemical evidence for the presence of an associated helper virus was found in the OK10-BM virus population produced by the macrophage cell line. However, when OK10-BM virus was serially passaged in chicken embryo fibroblasts, a virus having structural properties similar to those of OK10 AV (OK10 AV-specific oligonucleotides and gp85) appeared after three passages. Moreover, nonproducer clones of transformed cells could be readily obtained in OK10-BM virus-infected quail cell cultures. It is thus likely that the bone marrow-derived macrophage cell line produces a transforming virus defective in its env gene and low amounts of an associated helper virus, which upon transfer to fibroblasts is preferentially replicated. PMID- 6275103 TI - Isolation and characterization of a new bacteriophage, Cp-1, infecting Streptococcus pneumoniae. AB - Several pneumococcal phages showing a morphology completely different from those of all other previously found pneumococcal bacteriophages have been isolated. Bacteriophage Cp-1, one of the phages isolated, showed an irregular hexagonal structure and a short tail of 20 nm. The virion density was 1.46 g/cm3. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed the presence of nine polypeptides. The polypeptide showing a molecular weight of 39,000 accounted for more than the 90% of the total protein. The nucleic acid of Cp-1 was linear, double-stranded DNA with a mean length of 6.3 microns and a guanine-plus-cytosine content of 41%; its buoyant density was 1.699 and 1.422 g/cm3 in CsCl and CS2SO4, respectively. Its sedimentation coefficient (S20,w) was 19S. Cp-1 DNA showed a remarkable resistance to a large number of restriction endonucleases. A total of 12 fragments, ranging in molecular weight from 1.3 X 10(6) to 0.09 X 10(6), were produced by AluI, two fragments (molecular weight, 5.5 X 10(6) and 0.9 X 10(6)) were generated by HindIII, and two fragments (molecular weight, 6.0 X 10(6) and 5.7 X 10(6)) were produced by HaeIII. The easy visualization of th plaques produced by Cp-1, the small size of Cp-1 DNA (12 X 10(6) daltons), and other biological and physiochemical properties make this phage potentially useful for genetic studies. PMID- 6275104 TI - Theiler's murine encephalomyelitis virus group includes two distinct genetic subgroups that differ pathologically and biologically. AB - The intracellular development and RNA composition of Theiler's murine encephalomyelitis virus (TMEV) isolates were determined by electron microscopy, sucrose gradient centrifugation, and RNase T1 fingerprinting. Replication of FA virus, a virulent strain of TMEV, was characterized by the appearance of viral crystalline arrays in the cytoplasm of infected cells. In contrast, cells infected with the less virulent isolates (WW, TO4, BeAn 8386, and Yale) showed no crystalline arrays; instead, virions were found to be arranged between two layers of membranes in the cytoplasm of infected cells. Analysis of the RNAs of TMEV isolates showed that the RNAs were single-stranded molecules having sedimentation coefficients of 35S. RNase T1 fingerprinting of TMEV RNA revealed that striking differences between the virulent and less virulent TMEV isolates existed. Moreover, base composition analysis of RNase T1-resistant oligonucleotides of two TMEV isolates which represented the two subgroups indicated that there were no substantial oligonucleotides common to both subgroups. Based on these findings and the known difference in virulence, we suggest that the TMEV group contains two genetically district subgroups of viruses. PMID- 6275105 TI - Intracellular vesicular stomatitis virus leader RNAs are found in nucleocapsid structures. AB - Previous studies demonstrated that cytoplasmic extracts of cells infected with vesicular stomatitis virus contain plus-strand leader RNAs which sediment at 18S on sucrose gradients as a complex with viral N protein. The work presented in this paper demonstrated that these 18S complexes were stable on CsCl density gradients, banding at a buoyant density near that of genome nucleocapsids, and exhibited a morphology in an electron microscope similar to the disk structures found in virus genome nucleocapsids. Minus-strand leader RNAs were also found in 18S complexes on sucrose gradients. Quantitation of intracellular leader RNA suggested that, late in infection, approximately three-quarters of total intracellular leader RNA was encapsidated. PMID- 6275106 TI - Abelson virus-infected cells can exhibit restricted in vitro growth and low oncogenic potential. AB - We have designed a method for growing bone marrow cells infected with Abelson murine leukemia virus which permits examination of target cell growth early after infection. This culture system increases the efficiency of target cell growth by favoring rapid growth of a mixed population of adherent cells in the primary culture. The nonadherent Abelson virus-infected cell populations expressed pre-B cell differentiation markers characteristic of Abelson virus-transformed cells (mu-heavy chains of immunoglobulin M and terminal deoxynucleotidyltransferase). Early after infection, these cell populations exhibited restricted in vitro and in vivo growth properties which differed from those of an established Abelson virus-transformed cell line, 2M3. These included a marked dependency upon the adherent cell layer for growth and viability, a lower efficiency of agar colony formation, and a lower capacity for tumor production in syngeneic animals. Growth of the early populations could be maintained in the absence of the adherent cell layer by using conditioned medium from long-term adherent cell cultures established in the absence of viral infection. After passage of the populations for several weeks, the in vitro growth properties gradually shifted toward that of the 2M3 cell line. Twelve-week-old populations grew independently of the adherent cell layer and showed an increased efficiency of agar colony formation. These data indicate that many lymphoid target cells exhibit an intermediate transformed phenotype when infected with Abelson virus. Growth of these cells in culture is mediated via a synergistic interaction between intracellular expression of the viral transforming gene and an exogenous growth-promoting activity which can be provided by cultures of adherent bone marrow cells. PMID- 6275107 TI - Experiments with cloned complete tumor-derived bovine leukemia virus information prove that the virus is totally exogenous to its target animal species. AB - Taking advantage of the existence of a unique SacI restriction site in the long terminal repeats of the integrated bovine leukemia virus proviral DNA isolated from a bovine tumor, the total viral information (about 9.2 kilobases) was cloned in the lambdoid vector lambda WES. lambda B. Use of this cloned bovine leukemia virus DNA allowed us, for the first time, to definitely rule out the existence of any endogenous bovine leukemia virus sequence in the bovine, ovine, caprine, murine, feline, chicken, or human genomes. These data prove the absence of acquired cellular information in the provirus that has given rise to a tumor. PMID- 6275108 TI - Expression of adenovirus type 2 late genes in HeLa cells infected with adenovirus type 2 or adenovirus type 2-simian virus 40 hybrid viruses. AB - The relative levels of expression of the five 3'-coterminal mRNA families encode in the 16.4 to 99 map unit transcription unit of adenovirus type 2 (Ad2) are determined in part by the frequency of utilization of five possible polyadenylation sites. The possibility that polyadenylation frequencies at these sites may be regulated was tested by examining the expression of the fiber (polypeptide IV) gene in cells infected with nondefective (ND) Ad2-simian virus 40 hybrid viruses which express a sixth late mRNA family. Both the relative rate of fiber protein synthesis and the relative abundance of fiber mRNA were normal in cells infected with the hybrid viruses Ad2+ND1, Ad2+ND2, or Ad2+ND4. PMID- 6275109 TI - Cell surface T antigen in cells infected with simian virus 40 or an adenovirus simian virus 40 hybrid, Ad2+D2. AB - Cell surface T antigen, detected by a radioimmune assay that uses 125I-labeled Staphylococcus aureus protein A and antibodies against either authentic T antigen or D2 hybrid T antigen, was found in simian virus 40-transformed and -infected cells and in cells infected with an adenovirus-simian virus 40 hybrid, Ad2+D2. In simian virus 40 lytic infection, the surface T antigen appeared at the same time as the nuclear T antigen. PMID- 6275111 TI - Structure and phosphorylation of the Fujinami sarcoma virus gene product. AB - The Fujinami avian sarcoma virus (FSV) transforming gene product, P140, is a fusion protein which contains both gag-related and FSV-specific methionine containing tryptic peptides. The virion protease p15 cleaved p140 into two fragments: an N-terminal 33K fragment which contained all but one of the gag related tryptic peptides and a C-terminal 120K fragment which contained all of the FSV-specific tryptic peptides. The 33K gag-related fragment from P140 phosphorylated in FSV-transformed cells contained only phosphoserine, whereas the 120K C-terminal FSV-specific fragments contained both phosphoserine and phosphotyrosine. P140 isolated from cells infected at the nonpermissive temperature with an isolate of FSV which is temperature sensitive for transformation had a normally phosphorylated 33K fragment, but a hypophosphorylated 120K fragment deficient in both phosphotyrosine and phosphoserine. When P140 was immunoprecipitated from cells and phosphorylated in vitro at tyrosine residues in the immune complex kinase reaction, only the FSV specific fragment was labeled. These data define the structure of FSV P140 and locate the phosphorylated amino acids within the two regions of the polypeptide. PMID- 6275110 TI - Mutant carrying deletions in the two simian virus 40 early genes. AB - We isolated a simian virus 40 mutant, dl2194, which carried deletions in both early genes. One deletion removed 234 base pairs in the 54/59 region within the small-t-antigen coding sequence and the large-T-antigen gene intron. The second deletion removed 57 base pairs at the C terminus of the large-T-antigen coding sequence (0.20 map unit). dl2194 was a viable mutant, it carried a normal helper function for adenovirus growth on monkey cells, and it displayed the transformation properties of a small-t-antigen-negative single mutant. Therefore, none of the known large-T-antigen functions seemed to be altered by the C terminal deletion. PMID- 6275112 TI - Dexamethasone induction of the intracellular RNAs of mouse mammary tumor virus. AB - We have studied the kinetics of dexamethasone induction of mouse mammary tumor virus (MMTV) RNAs and proteins in virus-infected rat XC cells and GR mouse mammary tumor cells. A detectable increase in viral RNA in infected XC cells was present within 10 min after hormone addition, and half-maximal induction was achieved in less than 2 h. The increase in viral RNA concentration was apparent first in nuclear RNA and later in the cytoplasm. Within the first 15 min of induction, only genome-sized RNA (35S, 7.8 kilobases) was present in augmented amounts, whereas the major subgenomic RNA (24S, 3.8 kilobases) did not appear until at least 30 to 60 min postinduction. The sequential appearance of these RNAs, the probable mRNA's for the gag and env proteins, paralleled the order of appearance of the gag and env proteins, respectively, after hormone treatment. An additional species of viral RNA (20S, 2.5 kilobases) was detected during these induction experiments, but the role of this RNA is not known. Both subgenomic RNAs contain sequences derived from both the 5' and 3' termini of genomic RNA and are presumably spliced. After dexamethasone induction of infected XC cells, we detected two smaller env-related proteins which were not found in full hormone induction. The functional role of these smaller proteins is not known. A previously reported smaller species of RNA (13S, 1.0 kilobase) did not appear to be induced and was shown to be cellular rather than viral in origin. In the fully induced infected XC and GR mammary tumor cells, the only viral RNAs present were the 35S and 24S RNAs. In addition, mammary tumors contained only these two viral RNAs. Thus, tumor cells appear to contain only the viral RNAs which direct the synthesis of the gag, pol, and env proteins of the virion. PMID- 6275113 TI - Comparisons of rotavirus polypeptides by limited proteolysis: close similarity of certain polypeptides of different strains. AB - The polypeptides of SA11 rotavirus produced in virus-infected cells were analyzed by limited proteolysis, using Staphylococcus aureus V8 protease. This clearly distinguished between all the known primary gene products of the virus and allowed relationships between other infected-cell proteins, and between infected cell and virus structural proteins, to be ascertained. A comparison of the proteolysis cleavage patterns between SA11 rotavirus and the human rotavirus Wa was also performed which demonstrated a marked conservation in the digestion patterns among nonstructural and inner-shell structural proteins, but a marked variation in the digestion patterns among outer-shell structural proteins. PMID- 6275114 TI - Ultraviolet irradiation inhibits encapsidation of simian virus 40 chromatin. AB - During normal maturation and majority of pulse-labeled simian virus 40 DNA progresses from chromatin to previrions and virions within 5 h. UV light inhibits this progression. In heavily irradiated cultures (108 J m-2) most of the simian virus 40 DNA synthesized immediately before irradiation remains as chromatin for at least 5 h. This inhibition of maturation seems to be a result of the inhibition of protein synthesis. The data suggest that the pool of proteins required for maturation is sufficient to convert one-third of the simian virus 40 DNA molecules labeled in a 10-min pulse (at 33 h postinfection) from chromatin to previrions and virions and is exhausted within 1 h. PMID- 6275115 TI - Genetic mapping of endogenous mouse mammary tumor viruses: locus characterization, segregation, and chromosomal distribution. AB - The restriction endonuclease EcoRI has been used to study the inheritance of strain difference in endogenous mouse mammary tumor virus DNA sequences. This enzyme, which cleaves at only one site within the nondefective viral genome, generates DNA fragments containing mouse mammary tumor virus sequences which vary in size according to the locations of EcoRI restriction sites in the flanking mouse sequences, thereby defining unique integration sites of the viral genome. Recombinant inbred strains of mice have been used to study the inheritance of these DNA fragments which hybridize to mouse mammary tumor virus cDNA sequences. The results define 11 segregating units consisting of 1 or 2 fragments. These units were shown to segregate among the recombinant inbred strains, and in some instances linkage was established. Two units were shown to be linked on chromosome 1. Another unit was mapped to chromosome 7, which is presumably identical to the previously defined genetic locus Mtv- 1. One other mouse mammary tumor virus locus was tentatively assigned to chromosome 6. The results are consistent with the view that integration of mouse mammary tumor virus can take place at numerous sites within the genome, and once inserted, these proviruses appear to be relatively stable genetic entities. PMID- 6275116 TI - Host Susceptibility to endogenous viruses: defective, glycoprotein-expressing proviruses interfere with infections. AB - Three defective endogenous avian leukosis viruses, ev3, ev6 and ev9, interfered with subgroup E virus infections, ev3, ev6, and ev9 expressed high levels of subgroup E envelope glycoproteins. These glycoproteins reduced the activity of cellular receptors for subgroup E viruses. ev3 and ev6 protected chickens and cultured cells from subgroup E virus infections. PMID- 6275118 TI - Gene products of McDonough feline sarcoma virus have an in vitro-associated protein kinase that phosphorylates tyrosine residues: lack of detection of this enzymatic activity in vivo. AB - The primary translational product of the McDonough (SM) strain of feline sarcoma virus (FeSV) is a 180,000-dalton molecule, SM P180, that contains the p15-p12-p30 region of the FeLV gag gene-coded precursor protein and a sarcoma virus-specific polypeptide. In addition, cells transformed by SM-FeSV express a 120,000-dalton molecule, SM P120, that is highly related to the non-helper virus domain of SM P180. Both SM-FeSV gene products were found to be intimately associated with the membrane fraction of SM-FeSV-transformed cells. Immunoprecipitates containing SM P180 and SM P120 exhibited a protein kinase activity capable of phosphorylating tyrosine residues of both viral gene products but not immune immunoglobulin G molecules. By independently immunoprecipitating each of the two SM-FeSV proteins we found that most of the tyrosine-specific phosphorylating activity was associated with the SM P120 molecule. In vivo analysis of 32P-labeled SM P180 and SM P120 revealed their phosphoprotein nature; however, both molecules exhibited low levels of phosphorylation and did not contain phosphotyrosine residues. Finally, we did not detect any significant elevation in the levels of phosphotyrosine in the protein fraction of SM-FeSV transformants. Thus, if SM FeSV were to induce malignant transformation by a mechanism involving phosphorylation of tyrosine residues, the viral gene products must interact with a small subset of cellular proteins that do not represent a significant fraction of the total cellular protein content. PMID- 6275117 TI - Characterization of reticuloendotheliosis virus strain T DNA and isolation of a novel variant of reticuloendotheliosis virus strain T by molecular cloning. AB - Reticuloendotheliosis virus strain T (REV-T) is a highly oncogenic avian retrovirus which causes a rapid neoplastic disease of the lymphoreticular system. Upon infection, this virus gives rise to two species of unintegrated linear viral DNA, which are 8.3 and 5.5 kilobase pairs long and represent the helper virus (REV-A) and the oncogenic component (REV-T), respectively. Restriction endonuclease cleavage maps of these two DNA components indicate that REV-T DNA has a large portion of the genome deleted with respect to REV-A DNA and a substitution about 0.8 to 1.5 kilobase pairs long that is unrelated to REV-A DNA. These additional sequences comprise the putative transforming region of REV-T (rel). A chicken spleen cell line transformed by REV-T produced virus which upon infection gives rise to three species of unintegrated linear viral DNA (8.3, 5.5, and 3,3 kilobase pairs). We isolated the proviruses of the 8.3- and 3.3-kilobase pair species from this cell line by cloning in the phage vector Charon 4A. Restriction enzyme mapping showed that the two proviral clones are proviruses of REV-A and a variant of REV-T, respectively. A subclone of the variant REV-T provirus specific for the rel sequences of REV-T was used as a hybridization probe to demonstrate that the rel sequences are different from the putative transforming sequences of Schmidt-Ruppin Rous sarcoma virus strain A, avain myelocytomatosis virus, avian myeloblastosis virus, avian erythroblastosis virus, Abelson murine leukemia virus, and Friend erythroleukemia virus. In addition, the rel-specific hybridization probe was used to identify a specific set of sequences which are present in uninfected avian DNAs digested with several restriction enzymes. The corresponding cell sequences are not arranged like rel in REV-T. PMID- 6275119 TI - Genetic recombination of bacteriophage T7 in vivo studied by use of a simple physical assay. AB - A new physical method was developed to assay genetic recombination of phage T7 in vivo. The assay utilized T7 mutants that carry unique restriction sites and was based on the detection of a new restriction fragment generated by recombination. Using this assay, we reexamined the genetic requirements for recombination of T7 DNA. Our results were in total agreement with previous findings in that recombination required the products of genes 3 (endonuclease), 4 (primase), 5 (DNA polymerase), and 6 (exonuclease). Recombination was found to be independent of DNA ligase and DNA packaging and maturation functions. PMID- 6275120 TI - Delayed appearance of pseudotypes between vesicular stomatitis virus influenza virus during mixed infection of MDCK cells. AB - In intact Madin-Darby canine kidney (MDCK) cell monolayers, vesicular stomatitis virus (VSV) matures only at basolateral membranes beneath tight junctions, whereas influenza virus buds from apical cell surfaces. Early in the growth cycle, the viral glycoproteins are restricted to the membrane domain from which each virus buds. We report here that phenotypic mixing and formation of VSV pseudotypes occurred when influenza virus-infected MDCK cells were superinfected with VSV. Up to 75% of the infectious VSV particles from such experiments were neutralized by antiserum specific for influenza virus, and a smaller proportion (up to 3%) were resistant to neutralization with antiserum specific for VSV. The latter particles, which were neutralized by antiserum to influenza A/WSN virus, are designated as VSV(WSN) pseudotypes. During mixed infections, both wild-type viruses were detected 1 to 2 h before either phenotypically mixed VSV or VSV(WSN) pseudotypes. Coincident with the appearance of cytopathic effects in the monolayer, the yield of pseudotypes rose dramatically. In contrast, in doubly infected BHK-21 cells, which do not show polarity in virus maturation sites and are not connected by tight junctions, VSV(WSN) pseudotypes were detected as soon as VSV titers rose to the minimum levels which allowed detection of pseudotypes, and the proportion observed remained relatively constant at later times. Examination of thin sections of doubly infected MDCK monolayers revealed that polarity in maturation sites was preserved for both viruses until approximately 12 h after inoculation with influenza virus, when disruption of junctional complexes was evident. Even at later periods, the majority of each virus type was associated with its normal membrane domain, suggesting that the sorting mechanisms responsible for directing the glycoproteins of VSV and influenza virus to separate surface domains continue to operate in doubly infected MDCK cells. The time course of VSV(WSN) pseudotype formation and changes in virus maturation sites are compatible with progressive mixing of viral glycoproteins at either intracellular or plasma membranes of doubly infected cells. PMID- 6275121 TI - Expression of Epstein-Barr viral early antigen in monolayer tissue cultures after transfection with viral DNA and DNA fragments. AB - Antigens associated with the Epstein-Barr virus (EBV) replicative cycle were found in the nucleus and cytoplasm of human placental, Vero, BSC-1, and owl monkey kidney cells transfected with EBV DNA prepared from several different strains of virus. The number of antigen-positive nuclei increased when transfection was followed by cell fusion induced by inactivated Sendai virus. About 1,200 antigen-positive foci were induced per micrograms of EBV DNA. On the basis of their reactivity with various well-characterized human sera, it appears that the antigens are part of the early antigen complex. None of the four restriction endonucleases, EcoRI, HindIII, SalI, and BamHI, destroyed the ability of EBV DNA to induce early antigen. However, only SalI seemed to leave intact the full spectrum of antigen expression by the HR-1 and FF41 strains of EBV DNA. By means of transfection with recombinant DNA plasmids containing different EBV (FF41) DNA fragments regenerated by EcoRI, we showed that the coding region for early antigen was at least partially contained on the 17.2-megadalton EcoRI B fragment. PMID- 6275122 TI - Molecular genetics of herpes simplex virus. V. Characterization of a mutant defective in ability to form plaques at low temperatures and in a viral fraction which prevents accumulation of coreless capsids at nuclear pores late in infection. AB - In herpes simplex virus-infected cells, coreless capsids accumulate at the nuclear pores soon after infection, but subsequently disappear, suggesting that, as in adenovirus-infected cells (S. Dales and Y. Chardonnet, Virology 56:465-483, 1973), the release of viral DNA from nucleocapsids takes place at the nuclear pores. A nonlethal mutant, HSV-1(50B), produced by mutagenesis of HSV DNA fragments and selected for delayed production of plaques at 31 degrees C, accumulated coreless capsids at the nuclear pores late in infection in contrast to wild-type viruses. Recombinants selected for ability to produce plaques at 31 degrees C by marker rescue with digests of herpes simplex virus 2 DNA and selected clone fragments of HSV-1 DNA no longer accumulated empty capsids at nuclear pores late in infection. These results suggest that herpes simplex viruses encode a function which prevents accumulation of coreless capsids at nuclear pores, presumably by preventing uptake, unenvelopment, and DNA release from progeny virus, and indicate that the cold sensitivity of plaque formation and accumulation of coreless capsids might be related or comap in the S component of the genome. PMID- 6275123 TI - Characterization of a 70S polyuridylic acid polymerase isolated from foot-and mouth disease virus-infected cells. AB - A polyuridylic acid polymerase complex isolated from foot-and-mouth disease virus infected cells sedimented at 70S in a sucrose gradient and appeared in the exclusion volume of an agarose column whose molecular weight cutoff was 5 x 10(6). Phenol extraction of the complex yielded a heterogeneous band of virus specific RNA and an apparently host cell-derived 4.5 to 5S RNA, both of which are essentially single stranded. Neither RNA served as a template in the cell-free enzyme reaction. Polyacrylamide gel analysis revealed five polypeptides with molecular weights of 50,000, 56,000, 60,000, 70,000, and 74,000 and with molar ratios of 1:2:2:1:1, respectively. Autoradiography showed P56 to be the only major virus-induced polypeptide; the other proteins are apparently of host cell origin. Electron microscopic examination suggested a cartwheel shape for the polymerase complex which was seen to dissociate as polyadenylic acid was added. Antibody previously shown to inhibit enzyme activity aggregated the 70S units. PMID- 6275124 TI - Genetic relationship between the Mus cervicolor M432 retrovirus and the Mus Musculus intracisternal type A particle. AB - The genetic relationship between the retrovirus-like intracisternal type A particle (IAP) from Mus musculus and the novel retrovirus (M432) from M. cervicolor has been determined by heteroduplex and restriction endonuclease analyses of molecular clones of the respective genomes. We have found a major homology region (3.7 kilobase pairs) which probably begins near the 3' end of the M432 gag gene, spans the pol gene, and ends in the env gene. A second region (0.6 kilobase pairs) of weak homology was also observed adjacent to the 3' long terminal repeats of the respective genomes. The IAP genome is well conserved in the cellular DNA of all species of the genus Mus. In contrast, cellular DNA sequences related to the 5' end of the M432 genome, which shares no homology with the IAP genome, are found only in M. cervicolor and the closely related species M. cookii. These results suggest that the infectious M432 retroviral genome arose as a result of a recombinational event(s) between the IAP genome and another, as yet unidentified, class of retrovirus-related sequences or other cellular sequences. PMID- 6275125 TI - Properties of an intracisternal A-particle-associated endonuclease activity which is stimulated by ATP. AB - An endonuclease activity associated with purified proteinase K-treated intracisternal A-particles was identified and characterized. The activity required divalent cations, preferring Mn2+ to Mg2+. Salt concentrations above 50 mM inhibited the activity. The endonuclease was greatly stimulated by ATP, ADP, and dATP, whereas AMP appeared to produce a slight inhibition. GTP had no apparent effect on the activity. The enzyme introduced single-stranded nicks into DNA and nicked preferentially supercoiled DNA duplexes in the presence of ATP, although linear duplexes also functioned as substrates. Single-stranded DNA was not nicked to any great extent. The molecular weight of the enzyme was estimated to be about 40,000. The characteristics of this enzyme are very similar to those of the endonuclease found associated with Friend murine leukemia virus. PMID- 6275126 TI - Molecular cloning of viral DNA from human genital warts. AB - The DNA of human papilloma virus type 6 (HPV 6) has been cloned in Escherichia coli K-12 by using pBR322 as vector. The DNA was cloned at the BamHI and EcoRI cleavage sites. This DNA was mapped by employing further restriction endonucleases and by terminal labeling. No major differences were noted as compared to HPV 6 DNA originating directly from a genital wart. The existence of at least two DNA subtypes (HPV 6a and 6b) became apparent. PMID- 6275127 TI - Acyclovir-resistant mutants of herpes simplex virus type 1 express altered DNA polymerase or reduced acyclovir phosphorylating activities. AB - The biochemical properties of four acyclovir-resistant mutants are described. Two of these mutants, PAAr5 and BWr, specified nucleotidyl transferase (DNA polymerase) activities which were less sensitive to inhibition by acyclovir triphosphate than their wild-type counterparts. Another mutant, IUdRr, exhibited reduced ability to phosphorylate acyclovir. The fourth mutant, ACGr4, both induced an altered DNA polymerase and failed to phosphorylate appreciable amounts of acyclovir. BWr, a new acyclovir-resistant mutant derived from the Patton strain of herpes simplex virus type 1, induced a DNA polymerase resistant to inhibition by acyclovir triphosphate, but, unlike the polymerases induced by PAAr5 and ACGr4, still sensitive to phosphonoacetic acid. Resistance of BWr to acyclovir mapped close to the PAAr locus and was separable from mutations in the herpes simplex virus thymidine kinase gene by recombination analysis. PMID- 6275128 TI - Cleavage of the capsid protein precursors of encephalomyocarditis virus in rabbit reticulocyte lysates. AB - The cleavage pathway of the three capsid protein precursors of encephalomyocarditis virus, proteins A1, A, and B, was studied in rabbit reticulocyte lysates. Kinetic data suggested that the three proteins were cleaved through a sequential of A1 to A to B. PMID- 6275129 TI - Suppression of vesicular stomatitis virus defective intefering particle generation by a function(s) associated with human chromosome 16. AB - Human-mouse somatic cell hybrids were made between adenine phosphoribosyltransferase-deficient mouse L cells and a strain of human primary fibroblasts and selected in medium containing alanosine and adenine (J. A. Tischfield and F. H. Ruddle, Proc. Natl. Acad. Sci. U.S.A. 71:45-49, 1974). These hybrids were tested for the generation of defective interfering (DI) particles of vesicular stomatitis virus to determine whether or not a host gene controls the induction of DI particles. None of the seven independently arising hybrid clones tested generated detectable DI particles during 13 successive undiluted passages. In addition, the parental human cells also failed to generate DI particles. In contrast, the parental mouse cells generated a detectable level of DI particles during continuous passage. Thus, failure to generate DI particles appears to act in a dominant fashion in these hybrids. Human chromosome 16 and adenine phosphoribosyltransferase were present, as a direct consequence of the selection system, in all of the hybrid clones that failed to generate DI particles. It was the only human chromosome observed in the cells of every hybrid clone. This was verified by both isozyme and karyotype analyses. After hybrids were back-selected (with 2,6-diaminopurine) for loss of human adenine phosphoribosyltransferase and chromosome 16, they gained the ability to generate DI particles. Replication of DI particles already present in virus stocks, however, was normal in all of the hybrid clones and the parental human cells. This suggests that the induction, but not the replication, of DI particles is affected by the human genome and that a factor on human chromosome 16 seems to selectively suppress the mouse cell's ability to generate DI particles in the hybrids. These results support the idea that the induction of DI particles is controlled in part by host cell function(s), as suggested previously (C. Y. Kang and R. Allen, J. Virol. 25:202 206, 1978). PMID- 6275130 TI - Comparison of the restriction endonuclease maps of unintegrated proviral DNAs from four xenotropic murine leukemia viruses. AB - From purified linear and superhelical DNAs, the restriction endonuclease maps of four xenotropic murine leukemia virus DNAs from NFS, NZB, BALB/c, and AKR mice were determined with ten restriction endonucleases. Each xenotropic proviral DNA was found to be a unique restriction endonuclease map, with differences in the gag, pol, env, and terminal repeated sequence regions. However, type-specific SacI and EcoRI sites in the env region were identical in all four xenotropic murine leukemia virus DNAs and were not found in ecotropic murine leukemia virus DNA. Comparison of the xenotropic murine leukemia virus DNA maps with maps of ecotropic murine leukemia virus DNA showed that the pol and terminal repeated sequence regions were highly conserved. Other similarities in ecotropic and some xenotropic viral DNAs suggest common origins. PMID- 6275131 TI - Env gene products of AKR dual-tropic viruses: examination of peptide maps and cell surface expression. AB - The env gene products of nine AKR dual-tropic murine leukemia viruses were compared by peptide mapping and were assayed for expression on the cell surface of infected fibroblasts. Seven virus isolates expressed the env gene polyprotein on the cell surface. The env gene products of six of the seven viruses had identical peptide maps. The analysis of structure and expression of env gene products carried out in this study characterizes a subset of dual-tropic murine leukemia viruses shown by others to be thymotropic. PMID- 6275132 TI - Cloning overlapping DNA fragments from the B95-8 strain of Epstein-Barr virus reveals a site of homology to the internal repetition. AB - Overlapping, sheared DNA fragments from the B95-8 strain of Epstein-Barr virus were cloned in Charon 4A. Eleven recombinant phages plus one recombinant plasmid contained all of the sequences found in B95-8 virion DNA. Analysis of recombinant DNA molecules revealed a previously undetected site of homology to the internal repetition found in Epstein-Barr virus DNA. This site was adjacent to or at a site which was unstable when the recombinant DNA was propagated as phage DNA in procaryotic hosts. PMID- 6275133 TI - Virilizing adrenal adenoma secreting testosterone. PMID- 6275135 TI - Electron microscopic study of cardiac lesions induced in rats by isoproterenol and by repeated stress. With suggestion that idiopathic cardiomyopathy may be a "disease of adaptation". AB - An electron microscopic study of cardiac lesions induced in rats by isoproterenol and by repeated stress is described, and a possible pathogenetic mechanism of idiopathic cardiomyopathy is proposed. Three types of cardiac cells could be roughly differentiated according to the structure and arrangement of myofibrils: 1) normo-type with normal structure and normal arrangement of myofibrils, 2) nondestroyed-degenerated-type in which myofibrils show normal structure and abnormal arrangement, and 3) destroyed-degenerated-type with destroyed myofibrils. Various degrees and frequency of mitochondrial changes (accumulation, variation of size, swelling, loss of cristae and myelin figure), increase of glycogen granules, dilation of sarcoplasmic reticulum or T-tubules and nuclear changes were seen in every type. These 3 types seemed to correlate well with the degree of myocardial beta-adrenergic stimulation by isoproterenol or repeated stress. Normo-types were extensively observed in hearts exposed to mild myocardial beta-adrenergic stimulation. Nondestroyed-degenerated-types and destroyed-degenerated-types were frequently observed in hearts exposed to excessive myocardial beta-adrenergic stimulation. These results show a similarity between cardiac lesions induced in rats by isoproterenol or repeated stress and those seen in human idiopathic cardiomyopathy, and suggest the possibility that idiopathic cardiomyopathy is a "Disease of Adaptation." PMID- 6275134 TI - Herpes simplex encephalitis. Clinical Assessment. AB - Continuing evaluations of antiviral agents for treatment of herpes simplex encephalitis (HSE) provided an opportunity to collect clinical data from 113 patients in whom the diagnosis was proved by viral isolation. Occurrence of HSE was in all ages and in both sexes and was nonseasonal. Characteristically, patients had behavioral changes, fever, confusion, speech disturbances, and, less frequently, seizures. The EEG was the most useful neurodiagnostic aid followed by technetium and computed axial tomographic scans. Employing a logistic regression model for variable selection, the diagnosis could be predicted by clinical findings and neurodiagnostic tests in 83% of the proved cases, but the evidence in 25% was falsely positive. There was evidence of localization by either clinical or neurodiagnostic assessment in all patients with proved disease. Among patients wtih negative findings for HSE, similar focal findings predominated in all but a few. The CSF and brain scans were normal in many patients with proved HSE. This extensive clinical experience in patients wtih diagnosis proved by viral isolation shows that diagnosis cna be confirmed only by brain biopsy. PMID- 6275136 TI - Encephalomyocarditis (EMC) virus myocarditis in DBA/2 mice. I. Acute stage. AB - Severe myocarditis was induced in inbred DBA/2 mice inoculated with M variant of EMC virus. Yellowish-white patches were seen on the surface of the ventricles and atria, and histologically, myocardial necrosis and calcification was evident on the fourth day after virus inoculation. Myocardial lesions appeared earlier and were more extensive than we observed in previous myocarditis induced by Coxsackie B viruses. Spontaneous perimyocardial lesions were observed in control DBA/2 mice, which were exclusively limited to the right ventricle; however, these lesions were different in localization from lesions observed in infected mice in which extensive myocardial lesions were noted. This animal model is considered to be excellent for studies on the pathogenesis and natural history of viral myocarditis. PMID- 6275137 TI - Studies on regression of atherosclerosis--role of lipid containers. AB - The appearance of the aortic wall of rabbits fed a lanolin containing diet was examined by scanning (SEM) and transmission electron microscopy (TEM) in order to clarify the mechanism of removal of lipids deposited in the aorta. SEM study showed that circulating leukocytes penetrated into the arterial wall in the early phase of the experimental atherosclerosis. In some specimens, spherical bodies with a diameter of 10-30 mu were observed along the cleavage of the marginal folds between the endothelial cells. TEM demonstrated that these bodies contained many lipid particles, and numerous active microvilli were seen on the body surface. In the deeper subendothelial space, villi of each body interdigitated. From these results, it is postulated that the circulating leukocytes penetrate under the endothelium and take up the deposited lipids. Then, they return to the blood stream, containing lipid rich particles. The deposited lipids in the atherosclerotic lesion would be removed by this process. We named this phenomenon as "exo-tissuesis with lipid containers". While lipids are believed to be removed from the atherosclerotic lesion by HDL, lecithin cholesterol acyltransferase and others at the molecular level, we suggest that lipid containers play important roles in the regression and prevention of atherosclerosis on a major scale cellular and tissue level. PMID- 6275138 TI - Intracellular Ca++ transients and relaxation in mammalian cardiac muscle. AB - Calcium sensitive photoprotein aequorin was micro-injected into the cells of cat and rat ventricular cardiac muscles. Light and tension were recorded from the injected muscle in order to clarify the relation between the intracellular Ca++ transient and relaxation. The time course of the falling phase of the light signal and the time course of the relaxation were measured under the various conditions, i.e., changes of stimulation frequency, [Ca++] o, and the administration of adrenaline and caffeine. High stimulation frequency accelerated the time course of the fall of light and the relaxation of tension. Caffeine prolonged the relaxation of tension and the falling phase of light. The time course of relaxation did not change substantially but the falling phase time course of the light was accelerated by increase of [Ca++] o and adrenaline. These results confirm that a fall in [Ca++] i always precedes relaxation but suggest that the decrease of [Ca++] i is not always the rate limiting step of the relaxation in the mammalian cardiac muscle. PMID- 6275139 TI - [Effects of dibutyryl cyclic AMP on hepatic energy metabolism during hemorrhagic hypotension (author's transl)]. PMID- 6275140 TI - [Creatinine phosphokinase and adenylate kinase activities in skeletal muscle of malignant hyperthermia susceptible patients (author's transl)]. PMID- 6275141 TI - [Antibacterial activity of mecillinam against E. coli isolated from urinary tract infections (author's transl)]. AB - We studied susceptibility of 100 strains of clinically isolated E. coli by disc method to mecillinam. When inoculum size on agar plate was adjusted to 10(6) cells/ml and 10(9) cells/ml, respectively 96% and 63% strains were judged sensitive. Thus, in case of mecillinam MIC value as well as susceptibility tested by disc method highly depend on inoculum size and in the hospital laboratory inoculum size on agar plate should be carefully controlled even if it will be troublesome for routine disc examination. As it is suggested that clinical effectiveness of mecillinam will correspond to the result of 10(6) cells/ml inoculum size, we think the inoculum size should be adjusted 19(6) cells/l during the laboratory disc study. PMID- 6275142 TI - [Clinical evaluation of cefotiam in internal medicine, with special reference to infectious disease associated with hematological disorders (author's transl)]. AB - Clinical evaluation of cefotiam (panspolin), a new cephem antibiotics, was performed in the infectious disease associated with hematological disorders and in the respiratory system. In hematological dis orders, 40% of good and 25% of fair results were obtained in clinical effects. In respiratory infections, however, 92% of good results were obtained. Opportunistic infection due to Gram negative bacilli are often experienced in patients with leukemia. Since cefotiam has sufficient bacteriocidal effects in broad spectrum, it would be a good therapeutic agent against infectious diseases associated with hematological disorders. PMID- 6275143 TI - [Clinical evaluation of the new type pivmecillinam tablet (contains 100 MG) on the treatment for female acute simple urinary tract infections (author's transl)]. AB - Clinical studies of the new type pivmecillinam tablet (contains 100 mg) was performed on 24 patients with female acute simple urinary tract infections, receiving 300 mg 3 times daily for 7 days orally. Causative organisms in the cystitis cases were isolated singly as E. coli in 18 cases, Staph. aureus in 1 case and Staph. epidermidis in 1 case. From only 1 case with cystitis, E. coli and Staph. epidermidis were isolated mixedly. Clinical efficacy of the new type pivmecillinam tablet was evaluated as excellent in all 21 cases with female acute simple cystitis and 3 cases with female acute pyelonephritis. Side effect was not observed in the all cases. PMID- 6275144 TI - Effect of sodium intake on the brain and the aortic angiotensin-converting enzyme activity in spontaneously hypertensive rat. AB - Effect of sodium intake on angiotensin-converting enzyme activity was studied in five areas of the brain (cerebral cortex, midbrain, striatum, thalamus and hypothalamus) and in subcellular fractions of the aorta (homogenate, mitochondria, microsomes and supernatant) in normotensive, spontaneously hypertensive, and stroke-prone spontaneously hypertensive rats. Angiotensin converting enzyme activity was significantly higher in the hypothalamic area than in the other areas of the brain in spontaneously hypertensive rat. The enzyme activity of subcellular fractions of the aorta showed an extremely high value in the supernatant in normotensive, spontaneously hypertensive, and stroke-prone spontaneously hypertensive rats. Sodium intake resulted in a marked decrease in the aortic converting enzyme activity, while it did in a significant rise of the enzyme activity in the midbrain area in spontaneously hypertensive rat, and in the midbrain and striatum areas in stroke-prone spontaneously hypertensive rat. It is likely therefore that sodium intake lowers the converting enzyme activity of the aorta. Increased converting enzyme activity of the brain in spontaneously hypertensive rats may play a possible role in hypertension induced by sodium intake. PMID- 6275145 TI - [EB virus receptor test]. PMID- 6275146 TI - [Clinicopathological studies in patients with hepatocellular carcinoma. --VII. Two discrete systems of prognosis forecasting as revealed by RI scanning-- (author's transl)]. PMID- 6275147 TI - [Granular cell tumor of the esophagus (report of a case) (author's transl)]. PMID- 6275148 TI - [Cytomegalovirus inclusion disease with ulcerative changes of gastrointestinal tract (author's transl)]. PMID- 6275149 TI - [Serum complement profile and its clinical significance in patients with hepatocellular carcinoma and liver cirrhosis (author's transl)]. PMID- 6275150 TI - Fatal herpesvirus tamarinus infection in cotton-topped marmosets (Saguinus oedipus). PMID- 6275151 TI - Renal imagings in the diagnosis of polycystic kidney disease. PMID- 6275152 TI - Evaluation of residual renal function and prediction of the prognosis of polycystic kidney disease by 99m Tc-dimercaptosuccinic acid (DMSA) renal scintigraphy. PMID- 6275153 TI - Spontaneous necrotizing arteritis and glomerulonephritis in mice. PMID- 6275154 TI - Metabolic heterogeneity of the nephron. PMID- 6275155 TI - [Immunochemotherapy in prostatic cancer (author's transl)]. PMID- 6275156 TI - [Outbreak and control of Sendai virus infection in mouse breeding colonies (author's transl)]. PMID- 6275157 TI - Equivalency of endothelial cell growth supplement to irradiated feeder cells in carcinogen-induced morphologic transformation of Syrian hamster embryo cells. AB - Endothelial cell growth supplement (ECGS), an extract of bovine neural tissue with growth-promoting properties for human endothelial and epithelial cells and for mouse BALB/c fibroblast-like cells, can be substituted for feeder cells in a quantitative 7-day Syrian hamster embryo cell colony in vitro model of carcinogenesis. Inclusion of 50 or 100 micrograms ECGS/ml medium throughout the 7 day growth period produced results equal to those obtained with feeder cells. The frequency and morphology of normal fibroblast colonies and carcinogen-induced morphologically transformed cell colony growth in the presence of ECGS were similar to those in the presence of feeder cells. A positive dose-response relationship in transformation by benzo[a]pyrene occurred. The frequency of transformed colonies following UV irradiation and treatment of the cells with the cocarcinogenic tumor promoter 12-O-tetradecanoylphorbol 13-acetate was greatly augmented, and lymphotoxin, a lymphokine with anticarcinogenic activity, reduced transformation. Thus ECGS can substitute for feeder cells in supporting in vitro transformation and eliminates a potential complex source of variability for studies where interaction(s) with feeder cells are a consideration. The mechanics of this model system was simplified, and its versatility for the study of physiologic, carcinogenic, and other pathophysiologic processes was broadened. PMID- 6275158 TI - Effect of thymectomy on development of Lucke renal adenocarcinomas in virus infected leopard frog tadpoles. AB - Thymectomy of very young leopard frog (Rana pipiens) larvae dramatically reduced in vitro lymphocyte responses to the nitrogen phytohemagglutinin-P and prolonged survival of Lucke tumor alloimplants. However, tumor incidence in premetamorphic or metamorphosing frogs that had received injections as embryos of Lucke tumor herpesvirus (LTHV) was the same in thymectomized and normal immunocompetent animals. This result suggests that T-cell-mediated cytotoxicity does not prevent development of Lucke tumors in immunocompetent larvae and that if tumor-specific transplantation antigens appear, they may induce tolerance rather than destructive immunity in normal LTHV-injected larvae. PMID- 6275159 TI - Rat mammary tumor classification: notes on comparative aspects. AB - Mammary tumors (170 spontaneous and 1,613 induced with 7,12 dimethylbenz[a]anthracene) in inbred SD rats were classified histologically. The neoplasms were divided into three main categories: fibroepithelial, epithelial, and connective tissue. In the fibroepithelial category, compound tumors showing a wide range of histologic structures with variation in the arrangement of both epithelial and connective tissue elements that differ among lobules occurred in 44 (25.9%) spontaneous and 1,027 (63.7%) induced neoplasms, whereas fibroadenomas occurred in 41 (24.1%) untreated and 175 (10.9%) treated rats. Uncommon fissured tumors were found in 1 (0.7%) untreated and 6 (0.4%) treated animals, whereas carcinosarcomas were found in only 10 untreated animals. In the epithelial category, tubular adenomas occurred in 48 (28.2%) untreated and 164 (10.2%) treated animals. Cystadenomas, duct papillomas, intraductal carcinomas, and anaplastic carcinomas were found less frequently. Adenoacanthomas occurred in only 12 (0.8%) of the treated animals. Among tumors with a fibrous component only, fibromas occurred in 22 (13%) untreated and 97 (6.0%) treated animals, Fibrosarcomas were less frequent, occurring in 4 untreated (2.4%) and 54 (3.4%) treated animals. PMID- 6275160 TI - [Angiography in the diagnosis of pancreatic insulinoma]. PMID- 6275161 TI - [Results of treatment of synovial sarcoma]. PMID- 6275162 TI - Techniques for studying the pharmacodynamic effects of cardiac glycosides on patients' own erythrocytes during glycoside therapy. AB - We have measured the effects of digoxin on the cation transport mechanisms of patients' erythrocytes during treatment with digoxin for atrial fibrillation and cardiac failure in sinus rhythm. The results show that during short-term treatment with digoxin there is occupation of erythrocytic cardiac glycoside receptors by digoxin with resultant inhibition of active cation transport. These effects correlate well with the patients' clinical responses to treatment. During long-term treatment, however, these effects are not seen, suggesting that there is pharmacological tolerance to the effects of digoxin. The clinical implications of these results are discussed. PMID- 6275165 TI - Apparent cerebral atrophic findings on cranial computed tomography in nephrotic children with steroid therapy and in patients of infantile spasms with ACTH therapy. PMID- 6275164 TI - Zinc concentrations in plasma and erythrocytes in digitalized patients. AB - Determination of zinc, sodium, potassium and magnesium in plasma and red blood cells were performed in 31 controls and 63 patients treated with digitalis. In digitalized patients sodium and zinc concentrations in red blood cells were significantly increased. The intraerythrocyte magnesium and potassium levels as well as all investigated electrolytes in plasma remained within the normal range. There was a close relationship between the increase of sodium and zinc content in red blood cells, indicating alterations in transmembrane transport mechanisms induced by digitalis therapy. PMID- 6275166 TI - Superoxide anion radical (O2) generation by adriamycin and anthraquinone analogues. PMID- 6275163 TI - [The problem of the cellular receptor for cardiac glycosides (author's transl)]. AB - This review concerns the Na+, K+ -ATPase as well as the Na+, K+ -pump in the intact membrane and the highly specific inhibition of this transport system by cardiac glycosides. The interaction between glycoside and enzyme and the regulation of the kinetics of glycoside binding by ATP, K+, Na+, Mg2+ and Ca2+ are described. Emphasis is placed on the significance of the Na+, K+ -pump as the pharmacological receptor for cardiac glycosides. The problem encountered and progress made in attempting to correlate the inotropic action of cardiac glycosides with the binding of these drugs to the heart muscle and with the inhibition of the Na+, K+ -pump are reported. Recent results concerning increases of the intracellular Na+ concentration which are obtained by a partial inhibition of the Na+, K+ -pump and which are followed by an elevation of the intracellular Ca2+ -activity are reviewed. The discovery of a digitalis-like endogenous activity corresponds to the high specificity of the receptor for cardiac glycosides. PMID- 6275167 TI - [The practice of the public health nurses]. PMID- 6275168 TI - [Activities of the public health nurse. Infant welfare center]. PMID- 6275169 TI - [The public health nurse in a psychosocial service]. PMID- 6275170 TI - [Home care services in the Fribourg district]. PMID- 6275172 TI - [Care of a woman with cancer]. PMID- 6275173 TI - [In 2 neighboring communities of Neuchatel]. PMID- 6275171 TI - [The Association of Home Care Services]. PMID- 6275174 TI - [The Lausanne Center of home care services in the Red Cross section of Lausanne]. PMID- 6275175 TI - [How do nurses and Red Cross auxiliaries work together?]. PMID- 6275177 TI - [The domain of infancy]. PMID- 6275176 TI - [Who is the Red Cross auxiliary?]. PMID- 6275178 TI - [The cancer patient at home]. PMID- 6275179 TI - [Against violence. Rawson Prison, Argentine]. PMID- 6275180 TI - [What you should know about the ICN]. PMID- 6275181 TI - [Los Angeles, 28 June to 3 July 1981. Enthusiasm]. PMID- 6275182 TI - [Los Angeles, 28 June to 3 July 1981. New professional satisfaction and new courage from Los Angeles]. PMID- 6275183 TI - [The long-term patient in the acute hospital]. PMID- 6275184 TI - [From "gentle birth" to gentle aging and dying]. PMID- 6275185 TI - [Early warning system as a management tool for the nursing service]. PMID- 6275186 TI - [Swiss education for operating room nursing]. PMID- 6275187 TI - [The significance of the nurse in the health care team]. PMID- 6275188 TI - [Year of the Handicapped]. PMID- 6275189 TI - [Rapid organization of the operating room in mass admissions]. PMID- 6275190 TI - [The steel throat]. PMID- 6275191 TI - [A nursing education project]. PMID- 6275192 TI - [Reasons for leaving of nursing assistants]. PMID- 6275193 TI - [Conflict, its psychosocial, relational and therapeutic implications]. PMID- 6275194 TI - [Ergosociotherapy]. PMID- 6275195 TI - ["Soins infirmiers" presents itself]. PMID- 6275196 TI - [The care milieu, bureaucratic and solitary]. PMID- 6275197 TI - [Regression phenomena in adults who find themselves again in a student situation]. PMID- 6275198 TI - [Various behavior patterns and their possible origin in adults in education]. PMID- 6275199 TI - [Technological, scientific progress and the status of today's patients]. PMID- 6275200 TI - ["Krankenpflege" presents itself]. PMID- 6275201 TI - [The patient as a human being among humans]. PMID- 6275202 TI - [Thesis on the fundamental concerns of the nursing profession]. PMID- 6275204 TI - [Kardex for the community health nurse]U. PMID- 6275203 TI - [Self determination of patients]. PMID- 6275205 TI - Immunocytochemical and immunoelectron microscopic studies on Mallory bodies. PMID- 6275206 TI - Interactions of vascular wall cells with collagen gels. AB - Culture properties of bovine aortic endothelial cells and bovine aortic smooth muscle cells have been examined in relation to collagen gels. Endothelial cells grown on collagen maintain typical monolayer morphology not only in relationship to the overlying medium but also with respect to the collagen substrate. Endothelial cells placed within a collagen matrix assume a mycelial pattern resembling that of the microvasculature. Overlayment of confluent endothelial cells with collagen induces separation of the cells, changes in morphology, and reinitiation of growth. These shape changes do not require protein synthesis, appear to be independent of fibronectin, are not inhibited by cytochalasin D, but are inhibited by colchicine. Endothelial cell growth can be reinitiated by collagen. This phenomenon appears to be related to a change in cell shape and perhaps to separation of cells at the intercellular junction. In contrast, smooth muscle cells plated on collagen infiltrate the gels and assume a spindle-like, elongated morphologic appearance. Overlayment with collagen does not alter smooth muscle cell shape. Migration into the collagen gels is significantly enhanced when cells are cultured in medium containing platelet-released products, independent of growth stimulation itself. Migration is accompanied by collagen gel degradation. Release of labeled collagen into the medium by smooth muscle cells and appearance of TCA fragments in collagenase assay suggest secretion of collagenase. In summary, endothelial cells, but not smooth muscle cells, are restricted to the surface of collagen gels. The ability of the smooth muscle cells to invade is stimulated by platelet products and may be related to the synthesis of a smooth muscle collagenase. PMID- 6275207 TI - A rapid procedure for the screening and quantitation of barbiturates, diazepam, desmethyldiazepam and methaqualone. AB - A procedure is presented for the concomitant identification and quantitation of certain commonly abused weak acid and natural drugs from whole blood. This procedure offers several advantages over other procedures in that: (A) a means of extraction of the drugs from blood using a very inexpensive source of diatomaceous earth (Celite 560) is presented, (B) the addition of three internal standards allows the quantitation of several drugs from blood with just one extraction of the blood sample, (C) the extracts are virtually free of any contaminants from biological sources or from biologically inert drug metabolites, and (D) the barbiturates and desmethyldiazepam are derivatized to enhance their limits of detection and reduce peak tailing frequently observed when using the nitrogen-phosphorous detector and gas chromatography (NPD/GC). Some 1000 positive driving-under-the-influence cases have been quantitated by this method. PMID- 6275208 TI - Detection of tetrahydrocannabinol in blood. PMID- 6275209 TI - Effect of hormones on cyclic AMP in dog pancreatic tissue and secretion. PMID- 6275210 TI - Synovial sarcoma of the abdominal wall: a case report and study of its fine structure. AB - A case of synovial sarcoma in the abdominal wall of a 50-year-old Iraq female is reported. The ultrastructure of the tumor has been studied. Eight other cases of synovial sarcoma in the abdominal wall which reported in the English literature up to 1978 have been reviewed. Synovial sarcomas in the abdominal wall tend to occur with a much greater frequency in females in contrast to such tumors in the extremities or the neck which tend to occur with a much greater frequency in males. No ultrastructure studies of abdominal wall synovial sarcomas could be found in the English literature. Ultrastructure characteristics of this tumor were similar to those arising in the usual sites and in the neck. However, the present study has indicated that the light and dark cells reported by earlier workers are not solely limited to the epitheloid areas but are also present in the spindle cell zones. PMID- 6275211 TI - Effect of alkali metal salts on Sarcoma I in A/J mice. AB - The chloride salts of lithium (Li+) and cesium (Cs+) were evaluated for their ability to influence the growth of Sarcoma I implants in A/J mice. The administration of daily doses of either 1 or 3 mEq/kg CsCl to these mice reduced the incidence and size of tumor implants. This effect was not apparent in animals receiving a smaller dose (0.5 mEq/kg) of the same drug. At the time of sacrifice the serum level of Cs+ in this latter group was approximately half that recorded in animals receiving the higher doses of CsCl. No effect on tumor incidence or rate of growth was observed in animals receiving different doses of LiCl. Because of the similarities that existed between cesium and potassium, it was postulated that the effect of cesium was due to alterations in the intracellular composition of the tumor cells. Also, the possible role of cytotoxic agents in potentiating the inhibitory effect of cesium on tumors was discussed. PMID- 6275212 TI - Phase II trial of extended indications for resection is small cell carcinoma of the lung. AB - Surgical resection offers distinct theoretical advantages as the "local" modality in treatment of Stage I and II small cell carcinoma of the lung. We have treated 10 such patients by initial resection since 1975; all survivors but one received adjuvant chemotherapy for the full course thereafter. One patient died of a pulmonary embolus; the other nine remain without evidence of disease from 7 to 69 months after resection. A trial was undertaken of extended indications for resection in selected patients with Stage III-M0 disease. Criteria for patient selection have been developed gradually; these exclude patients for reasons of refusal, physiological inadequacy, disease unsuited to gross total eradication, or lack of adequate initial response to chemotherapy. Of six patients who survived the exclusion criteria and underwent resection, one has had a relapse at 26 months. All others remain without evidence of disease, 5 to 25 months after the start of treatment. We believe that systematic patient selection on the basis of defined criteria will identify a subset of patients having markedly improved chances for disease control. This group may represent as many as half of the patients first presenting with localized or MO disease. Patients excluded as candidates for resection have continued to receive standard nonsurgical combined modality therapy. PMID- 6275213 TI - Pulmonary sarcoidosis and serum angiotensin-converting enzyme. PMID- 6275214 TI - A new method which investigates lysosomal sensitivity to superoxide with endocytotic, site-specific, chemiluminescent probes. AB - The end result of cellular autophagocytosis and lysosomal digestion is incompletely understood in regard to the roles of the superoxide anion radical (O2-.). Cultured glial cells sequestered endocytotically two probes that were site-specific to the lysosome vacuome and sensitive to free-radical activities. The Sepharose-4B-polyisoluminol probe emitted chemiluminescent light in proportion to externally injected O2(-).. Bioluminescence measurements of beta glucuronidase activity in intact glial cell lysosomes was achieved with the second site-specific, enzyme-specific Sepharose-4B-(dodecanate)5' luciferinylglucuronate probe. Considerable activity was found in the lysosomal vacuome. In conclusion, we suggest that the use of site-specific, chemiluminescent probes can be of importance in the study of free radicals. PMID- 6275215 TI - Application of chemiluminescent probes in investigating lysosomal sensitivity to superoxide versus suspected radical scavengers. AB - The role of the superoxide anion radical (O2(-).) relative to catalytic/inhibitory substances in lysosomes is poorly understood. Cultured glial cells sequestered endocytotically two probes that were site-specific to the lysosome vacuome and sensitive to radical activities. The Sepharose-4B-isoluminol probe emitted chemiluminescent light in proportion to externally injected O2(-). and was inhibited or catalysed by various radical scavengers, transition metals and other substances which may affect lysosomal metabolism and lipofuscin formation. We conclude that lysosomal radical activities may be inhibited by butylated hydroxytoluene, hydrocortisone, ACF, RNA, alpha-tocopherol, and, in special circumstances, with fully metabolized iron. Choline and oxidized copper and iron cations in overload concentrations in incubated freshly in lysosomes may catalyse radical activities, and they may be important factors in lipofuscin formation and its role in aging. PMID- 6275216 TI - An application of bioelectrical impedance for the detection of tissular bubbles in the diver: preliminary results. PMID- 6275217 TI - [Cerebral and pulmonary hemorrhage after lymphography (author's transl)]. PMID- 6275218 TI - Cytomegalovirus and the otolaryngologist. AB - Human cytomegalovirus (CMV) is ubiquitous and has been documented as the etiologic factor in several diseases ranging from respiratory tract infection to congenital deafness. Special clinical manifestations of CMV of interest to the otolaryngologist are presented as related to the temporal bone and audiovestibular system, the facial nerve, the aerodigestive tract, the thyroid, the hematopoietic-lymphoreticular system, and salivary gland. In addition, current concepts of CMV oncogenic potential are discussed as they relate to head and neck neoplasms. The results of a study performed on surgical pathology specimens obtained from 26 patients, in an attempt to isolate CMV in tissue culture from salivary gland and/or neoplasms of the head and neck, detected no evidence of viral presence. The significance of these findings and the role of future research is discussed. PMID- 6275219 TI - Clinical-pathological determinants in prognosis of fibrous histiocytomas of head and neck. AB - Review of the literature reveals 76 cases of fibrous histiocytoma of the deep structures of the head and neck. To this we have added 11 of our own cases, and analyzed the total group to determine clinical and histological features of a high risk population. Age, sex, presence of pain, location of tumor, size of tumor, and local invasion appear to be important clinical criteria. Bizarre giant cells, numerous and atypical mitoses, necrosis, and inflammation appear to be important histologic features. The treatment of choice is wide local excision. Chemotherapy appears to produce tumor regression. PMID- 6275220 TI - [Hepatoblastoma in adults (author's transl)]. AB - During autopsy a hepatoblastoma was found in a 73-year-old woman. The tumor contained fetal hepatocytes as an epithelial component and spindle-shaped cells resembling fibroblast as well as focal osteoid tissue representing mesenchymal components. Hepatoblastoma are predominantly found in children and are very rare in adults. Hepatoblastoma probably arise from multipotent blastema which can develop into epithelial and mesenchymal tissue. PMID- 6275221 TI - Effects of the intracerebroventricular injection of antinociceptive doses of acetylcholine on the stereospecific binding of 3H-dihydromorphine. PMID- 6275222 TI - The beta-adrenergic receptor in human lymphocytes: subclassification by the use of a new radio-ligand, (+/-)-125 Iodocyanopindolol. PMID- 6275223 TI - The (non)specificity in the lipid-requirement of calcium- and (sodium plus potassium)-transporting adenosine triphosphatases. PMID- 6275224 TI - Reduced number of adrenal angiotensin II receptors in the spontaneously hypertensive rat. PMID- 6275225 TI - The effects of hormonal and circadian cycles, stress, and activity on levels of cyclic AMP and cyclic GMP in pituitary, hypothalamus, pineal and cerebellum of female rats. PMID- 6275226 TI - Inhibitory effects of pergolide on peripheral adrenergic neurotransmission in spontaneously hypertensive rats. PMID- 6275227 TI - Identification of solubilised beta 1 and beta 2 adrenoceptors in mammalian lung. PMID- 6275228 TI - Central actions of valproic acid in man and in experimental models of epilepsy. PMID- 6275229 TI - The effect of beta-endorphin on arginine-induced growth hormone and prolactin release. PMID- 6275230 TI - Response of rhesus monkey lymphocytes to short-term administration of THC. AB - Four Rhesus monkeys were subjected to daily administration of 2.5 mg of tetrahydrocannabinol (THC)/kg body wt, after establishing the norms for complete blood count, T- and B-cell concentrations, and the dose response of thymidine incorporation after PHA stimulation. THC was administered daily for 3 weeks, the treatment was stopped, and then the animals were allowed to recover for 4 weeks. Cellular responses, incorporation studies and fibrinogen levels were determined during the treatment and recovery phases. Compared to 4 vehicle-treated animals, the THC-treated animals experienced significant augmentation of both their total white cell and their neutrophil counts during the recovery phase which returned to normal levels during the recovery phase. There was no alteration in total lymphocyte count or T- or B-cell concentrations. Fibrinogen levels of the THC treated animals during the treatment phase were also elevated compared to controls, and the levels diminished to the same values as the vehicle-treated animals during recovery phase. Possible mechanisms for the response of Rhesus monkeys to short-term administration of THC are discussed. PMID- 6275231 TI - Chronic haloperidol treatment increased calcium-dependent phosphorylation in rat striatum. AB - In previous studies, we observed that when rats were chronically treated wih haloperidol, there was a significant increase of calmodulin activity in their striatal membranes. Calmodulin is known to modulate calcium-dependent protein phosphorylation in neural membranes. In the present study, we found that the total 32P-incorporation in the striatal proteins from chronic haloperidol-treated rats was significantly increased in comparison to saline-treated rats. A majority of the phosphorylation was attributed to the calcium-mediated activity, since it could be blocked by a calcium chelating agent (EGTA). By using EGTA to inhibit phosphorylation, the results indicated that the haloperidol-treated rats had approximately 3.5-fold greater Ca++-dependent protein kinase activity than the saline-treated rats. Exogenous calcium alone was insufficient to stimulate phosphorylation in the haloperidol-treated rats to the same magnitude as in the saline-treated rats. Calmodulin may be required. 32P-incorporation of two striatal proteins at molecular weight 40 and 52 kilodaltons were markedly stimulated by calcium. Cyclic AMP-mediated phosphorylation seemed to take only a small part in the alteration of total phosphorylation. Therefore, the increase of calmodulin activity and calcium-dependent phosphorylation appears to play a major role in the drug-induced dopamine receptor supersensitivity in rat striatum. PMID- 6275232 TI - Ontogeny of alpha 1- and beta-adrenergic receptors in rat lung. AB - The binding characteristics of the alpha 1-selective adrenergic ligand [3H] prazosin were determined in particulate membranes of rat lung from day 18 of gestation to adulthood. Specific binding was present at all ages studied, was reversible and inhibition of specific binding by agonists followed the order of potency: (-)-epinephrine = (-)-norepinephrine much greater than (-)-isoproterenol greater than (+)-norepinephrine. Inhibition by antagonists followed the order of potency: prazosin greater than WB4101, much greater than yohimbine. Binding capacity increased during the neonatal period from 52 +/- 9 fmoles x mg-1 protein in lung preparations on day 18 of a 21 day gestation increasing to 105 +/- 4 fmoles x mg-1 protein (mean +/- SE) by postnatal day 15. Binding activity decreased thereafter, reaching adult levels by 28 days of postnatal age, 62 +/- 3 fmoles x mg-1 protein. This pattern of alpha 1-adrenergic receptor density was distinct from that of beta-adrenergic receptors identified in rat lung membrane with the beta- adrenergic antagonist, (-)-[3H]dihydroalprenolol ((-)-[3H]DHA). ( )-[3H]DHA binding increased dramatically during this same time period, from 46 +/ 4 fmoles x mg-1 protein on day 18 of gestation to 496 +/- 44 fmoles x mg-1 protein in the adult lung. Affinity for [3H]-prazosin and (-)-[3H]DHA did not change with age. Pulmonary alpha 1-adrenergic receptors are present as early as 18 days of gestation in the rat and alpha 1-adrenergic receptor density is maximal by 15 days of postnatal age. The timing of the changes in alpha 1 adrenergic receptors correlates with the timing of increased sympathetic innervation of the developing rat lung and is distinct from that of beta adrenergic receptor sites. PMID- 6275233 TI - A quantitative analysis of the shaking behavior induced in rats by beta-endorphin and [D-Ala2, Met5]enkephalinamide. AB - beta-Endorphin (5-80 microgram) or [D-Ala2,Met5]enkephalinamide (DALA) (5-40 microgram) was administered intracerebroventricularly to rats. With both opioid peptides, there was no direct relationship between log dose and mean number of wet-dog shakes (WDS) that occurred during the following 15 min. When the results were analyzed quantitatively, the dose of DALA that caused 50% of the rats to shake at least twice was 8.6 microgram (4.9-15 microgram). beta-Endorphin had such poor efficacy that an ED 50 could not be obtained. Morphine (1 and 5 mg/kg, s.c.) antagonized shaking caused by the optimal dose of DALA (20 microgram). Naloxone (0.1-10 mg/kg, s.c.) attenuated both DALA- and beta-endorphin-induced WDS in a dose-related manner. This latter result differentiates shaking associated with opioid peptides from that caused by thyrotropin releasing hormone (TRH), another endogenous stimulant of WDS in rats. There was no cross-tolerance between RX 336-M (7,8-dihydro-5',6'-dimethylcyclohex-5'-eno-1',2',8',14 codeinone), a novel shake inducing agent, and beta-endorphin. This finding again differentiates beta-endorphin-induced shaking from that caused by TRH and also from that associated with several exogenous stimulants of WDS. PMID- 6275234 TI - [Optimization of the dosage fields of a proton beam in treating cervical cancer]. PMID- 6275235 TI - Medical testimony in malpractice case arising out of total hip replacement surgery--sciatic nerve damage--discovery deposition of defendant orthopedic surgeon. PMID- 6275236 TI - [Clinical aspects of ocular onchocerciasis in Ivory Coast nine years after the interruption of the vectorial transmission (author's transl)]. AB - Report of 262 ocular controls in Ivorian adults nine years after they left their respective villages in hyperendemic onchocerciasic areas which were flooded consequently to the building up of the Kossou dam. After that period 50 p. 100 of the controlled persons have still a positive skin biopsy. On the other hand, parasitism by Onchocerca volvulus microfilariae and lesions of keratitis punctata have nearly disappeared (respectively 1 and 2 cases). Patients with microfilariae in the anterior ocular camera are very rare (3,4 p. 100) though many patients show definitive changes of the anterior and posterior ocular segments. Such kind of surveys are necessary to have a good evaluation of the regressive evolution of ocular schistosomiasis after the interruption of the vectorial transmission. PMID- 6275237 TI - [Immunologic changes in patients with Hodgkin's disease in clinical remission and recurrence]. PMID- 6275238 TI - Spin immunoassay. PMID- 6275239 TI - Membrane immunoassay: a spin membrane immunoassay for thyroxine. PMID- 6275240 TI - Specific radioimmunoassays for rabbit liver fructose bisphosphatase, pyruvate kinase, and glycerol-3-phosphate dehydrogenase. PMID- 6275241 TI - Radioimmunoassay of bovine type II cAMP-dependent kinase. PMID- 6275242 TI - Antibodies to prolactin receptors and growth hormone receptors. PMID- 6275243 TI - Measurement of thyrotropin receptor antibodies. PMID- 6275244 TI - Production and application of an antibody specific for the cardiac beta adrenergic receptor. PMID- 6275245 TI - A hypothesis for the mechanism of superoxide production by phagocytic cells. AB - When arachidonic acid is reacted with Fe2+, the Fe2+ is oxidized to Fe3+, and lipid peroxides are formed as detected by absorption at 232 nm. However, there is a large discrepancy between the amount of arachidonic acid converted to stable oxidized products and the amount of Fe2+ converted to Fe3+. We suggest that under appropriate conditions, electrons from the ferrous iron may reduce oxygen to superoxide in the presence of arachidonic acid. The results are consistent with evidence using bromophenacyl-bromide suggesting that release of fatty acids from cell phospholipids might be required for phagocytes to synthesize superoxide. PMID- 6275246 TI - Cardiac nuclear medicine, part III: acute myocardial infarction and other applications. PMID- 6275247 TI - Blood pressure changes: oestrogens in climacteric women. AB - An analysis of data from a prospective study which compared the effect of conjugated equine oestrogens (Premarin) with that of piperazine oestrone sulphate (Ogen) demonstrated that postmenopausal women who received conjugated equine oestrogens had a higher rate of a rise in blood pressure and development of hypertension than women who received piperazine oestrone sulphate. Conversely, a greater proportion of women who took oestrone sulphate had a fall in blood pressure than those who took equine oestrogens. A hypothesis to account for these differences, based on the known potent biological action of equine oestrogen, is developed. PMID- 6275248 TI - Possible ranitidine hepatitis. AB - A 63-year-old woman developed anicteric hepatitis after two weeks of therapy with ranitidine. Despite continuation of therapy, her symptoms resolved within five days and transaminase levels returned to normal in the next four weeks, at which time use of the drug ceased. The gamma-glutamyl transpeptidase level returned to normal two weeks later. Although liver biopsy and drug rechallenge were not performed, no other cause for the reaction could be found. PMID- 6275249 TI - Bone marrow transplantation using fractioned total body irradiation: Absence of pulmonary toxicity despite cytomegalovirus viraemia. AB - Bone marrow transplantation was performed on a 22-year-old male with acute myeloid leukaemia in early relapse. The donor was his HLA-identical, but ABO incompatible, brother. Total body irradiation (TBI) was given in six daily doses of 2GY rather than as the usual single dose of 10GY. After transplantation, cytomegalovirus viraemia occurred, but the patient did not develop pneumonia despite the reported high association of these events. Four hundred days after transplantation, the patient was fit and active with no haematological abnormalities. The use of fractionated TBI may be associated with a lower incidence of respiratory complications than standard single-dose TBI. PMID- 6275250 TI - Hepatitis A virus in cell culture. II. Growth characteristics of hepatitis A virus in Frhk-4/R cells. AB - The propagation of hepatitis A virus (HAV) is shown in a rapidly growing fetal rhesus monkey kidney cell line (Frhk-4/R). In the course of ten passages through Frhk-4/R cells the HAV, which was released in the supernatants of the cell cultures, was adapted to these cells. In the tenth passage 6 days after infection, it was possible to detect low amounts of HAV in the supernatants by radioimmunoassay (RIA). With indirect immunofluorescence 3 days after infection, a specific granular fluorescence was demonstrated in the cytoplasm of cells infected with HAV from the tenth passage. A persistent infection of Frhk-4/R cells appeared with a main peak at a density of 1.32 g/cm(3) in CsCl density gradients. In the 1.32 g/cm(3) fractions, typical HAV particles are shown by electron microscopy. Furthermore the HAV produced in Frhk-4/R cells is shown to be a useful antigen for diagnostic tests. PMID- 6275251 TI - Hepatitis A virus in cell culture. III. Propagation of hepatitis A virus in human embryo kidney cells and human embryo fibroblast strains. AB - The propagation and adaptation of hepatitis A virus (HAV) in human embryo kidney cells (HKC) is shown. The growth curve of HAV in the first passage through HKC is compared to the growth curve in the tenth passage through HKC. It is shown that in the course of 18 passages through HKC, HAV adapted to these cells causing the virus to grow much more rapidly. The cell-bound HAV is compared to the HAV released in the cell-culture supernatant during the ninth passage through HKC. The HAV from the tenth passage through HKC is shown to be able to replicate also in a human embryo fibroblast strain (HFS). Furthermore, adaptation of the HAV to HFS is demonstrated. PMID- 6275252 TI - Complement-fixing antibodies to human cytomegalovirus induced early nuclear antigens in mononucleosis. AB - Early nuclear antigen (CMNA) induced by human cytomegalovirus (HCMV) was extracted from infected human embryonic fibroblast cells and purified by ds DNA Sephadex chromatography. The purified antigen was added to acid-fixed preparations from human embryonic fibroblasts and these in vitro converted nuclei were exposed to human sera to estimate the antibodies to CMNA by anti-complement immunofluorescence staining. Serial serum samples were obtained from eight patients suffering from acute HCMV infection (mononucleosis), from nine patients with Epstein-Barr virus (EBV) mononucleosis, and from 20 healthy persons who had been shown to possess antibodies to HCMV late antigens. Acute HCMV infection is characterized by the presence of anti-CMNA antibodies at high titer (1:16 - 1:32) together with the elevated level of IgG antibodies to anti-HCMV-late antigens. During convalescence the anti-CMNA titer decreased to a lower level which was maintained for long period. The anti-CMNA antibodies were also regularly detected in sera of persons possessing antibodies to HCMV late antigens but without any sign of acute HCMV infection. It is concluded that antibodies to CMNA are not transitory and their presence even at high titer (1:16 - 1:32) in a serum sample cannot be taken as a presumptive evidence of acute virus infection. This method seems to be valuable for measuring antibodies to nuclear DnA-binding antigens induced by other viruses. PMID- 6275253 TI - Improved measurement of neutron spectrum from 35 MeV protons on thick beryllium. PMID- 6275254 TI - "Battered data"--some clinical effects of the abuse of multiple regression methods: a second look at some observations on Ro07-0582 (Misonidazole). AB - The paper presents an argument for the wider use of simple and standard statistical methods in the design and evaluation of both laboratory and clinical experiments with toxic radiosensitizers. It also presents an argument against the frequent practice of superposing a curve estimated from laboratory observations upon scattergrams of clinical observations to thereby assist the reader to the (often fallacious) inference that the two disparate sets of observations have enough in common to be graduated by the same curve. Such a practice achieves a substitution of reasoning by analogy for the more reliable statistical procedures of estimation and of testing a hypothesis. PMID- 6275255 TI - [Sigmoid neoplasms in patients with ureterosigmoidostomy]. PMID- 6275257 TI - [Pyrophosphate analogs in the pyrophosphorolysis reaction catalyzed by Escherichia coli RNA polymerase]. AB - Processive pyrophosphorolysis of RNA from ternary RNA polymerase-nascent RNA delta D111 T7 DNA complex has been followed in the absence of nucleoside triphosphates. Series of inorganic pyrophosphate analogs were investigated for their ability to sustain the reaction and to compete with inorganic pyrophosphate for the reaction. Methylenediphosphonic, imidodiphosphonic, phosphonacetic acids, inorganic triphosphate, methylenediphosphonic and phosphate were found to be capable of substituting the inorganic pyrophosphate in RNA degradation reaction with tantamount efficiency. They give rise to nucleoside monophosphates for phosphonoacetic acid, nucleoside triphosphates for inorganic pyrophosphate and inorganic triphosphate, nucleoside triphosphates analogs for methylenediphosphonic, imidodiphosphonic acids and methylenediphosphonic acid phosphate as the low molecular weight product of the reaction. The problem of specific interaction of RNA polymerase with nucleoside triphosphates and inorganic pyrophosphate is discussed in the terms of structural requirements for the compounds to be a potent substrate for RNA polymerase. PMID- 6275256 TI - Ubiquitin - protein conjugates. AB - The data available at present indicates there are three distinct functions of ubiquitin, two of which are related to protein conjugation. The first of these has been extensively studied by our laboratory and others interested in nucleosomes and changes in chromatin states. The ubiquitin-histone (Ub-2A, Ub-2B) conjugation reaction now appears to be a very dynamic process. In the deconjugation (lyase) reaction, both the histone 2A and the ubiquitin are left intact and in a form which makes possible ready reconjugation. Accordingly, this may be a mechanism for "moment-to-moment" Control of the genome. The second function in which ubiquitin is conjugated involves proteolytic activity. This activity is correlated with protein turnover. In this process, the ubiquitin protein conjugate apparently serves as a "signal" for the protease cleavage of the protein. The released ubiquitin is also intact and is probably available for reconjugation. In the third function, ubiquitin was suggested to serve as a "hormone". The studies thus far have been carried out primarily on induction of T and B-lymphocytes, reduction or delay of Coombs' positivity and reduction of spleen weight. The precise physiological role of this reported function is still unclear, particularly because the ubiquitin used was probably not the physiologically active form. PMID- 6275258 TI - [Interaction of RNA polymerase with hybrid plasmids carrying Escherichia coli threonine operon]. AB - The RNA polymerase binding with two hybrid plasmids carrying threonine operon genes was studied. RNA polymerase binding sites were localized using nitrocellulose binding assay and electron microscopy visualization of the RNA polymerase--DNA complexes. To confirm that observed RNA polymerase binding sites are real promoters we analyzed RNA transcripts synthesized in vitro by hybridization with DNA fragments. The promoters of following genes were localized on the plasmid maps: ampicilline, threonine and RNA-primer of DNA replication. Two latter promoters bound RNA polymerase only being in the supercoiled DNA molecule. Several additional binding sites were found. The positions of some of these sites corresponded with known sites of rho-independent transcription termination. PMID- 6275259 TI - [Restriction mapping of plasmid pSa1, isolated from Streptomyces albus G strain]. AB - A novel plasmid designated pSa1 has been isolated from Streptomyces albus G strain producing SalGI restriction endonuclease. Molecular weight of the plasmid is 3.4 +/- 0.2 mD. The action of 12 restriction endonucleases on the plasmid DNA was studied. Restriction map of pSa1 DNA was established for SmaI, HindII, XbaI and KpnI endonucleases. PMID- 6275260 TI - [Molecular mechanisms of photoreception. V. Reconstitution of a sodium channel on an artificial lipid membrane modified by bovine rod outer segment components]. AB - An attempt is made to reconstitute the Na+-conductivity elements of rod outer segment (ROS) plasma membrane on the artificial lipid membrane (ALM). ALM were modified by preparation with bovine ROS plasma membrane fragments. Discrete fluctuations of the ALM conductivity were observed at addition of this preparation to the ALM bathing solutions to a final concentration of 0.1--1.0 microgram/ml. The magnitude of these fluctuations was about 25 pS at 140 mM NaCl. The modified ALM possessed preferentially the Na+-conductivity, which was at least five times as great as that for K+ or Li+. The modified ALM showed practically no conductivity for Cl-. The Na+-channels were voltage-dependent. The Na+-channels were "sensitive to visible light" at some experimental conditions. The optimal conditions were obtained for reconstitution of Na+-channel on the ALM. The temporal and termal stabilities of the modified preparation were investigated. It was shown that the frequency of the modified ALM conductivity fluctuations are dependent on temperature and on lipids composition of ALM. The data obtained in our work are in a good agreement with the results of electrophysiological studies of photoreceptor cells. It may be indicated, that the investigated ALM contains the Na+-channel of the ROS plasma membranes. It is suggested that the reconstituted system will be useful for studying the principles of regulation of ROS plasma membrane sodium conductivity and the nature of a mediator in a photoreceptor transduction mechanism. PMID- 6275261 TI - [Modification of guanine residues in double-stranded DNA by aminomethylol compounds without denaturation of nucleic acid]. AB - With the application of radioactive formaldehyde and glycine the ability of aminomethylol compounds to combine with S1 nuclease treated DNA at 25 degrees and pH 5.8--7.4 has been shown. The reaction leads to modification of 22--26% of base pairs without changes of the DNA UV-absorption spectrum. Besides that the flexibility coefficient, the kinetics of despiralization under the action of formaldehyde and the stability of DNA molecule towards the S1 nuclease action permit to conclude that modification does not cause DNA despiralization. In experiments with the use of synthetic double-stranded polynucleotides poly(dA) times poly(dT), poly(rC) times poly(rl), poly(rG) times poly(dC) and poly(dC-dG) times poly(dC-dG) it has been shown that binding of methylol compounds to nucleic acids is due to reaction with guanine residues. Methylol derivatives of glycine reacts with guanine residues of double-stranded DNA only 10 times slower than with the monomer--deoxyguanosine-5'-phosphate. The studied reaction is reversible and the half-period of modified DNA reduction is found to be 5 hours at 25 degrees and pH 6.5. The rate constants of forward and reverse reactions and equilibrium constants of the reaction between methylolglycine and native DNA were determined. PMID- 6275262 TI - [Electron microscopy of colonic polyps. II. Leuchtenberg's inclusion bodies]. AB - Characteristic features of Leuchtenberger's inclusion bodies of adenomatous, adenopapillary and papillary polyps of the colon were studied by electron microscope. Authors believe that in the development of intracellular inclusion bodies focal degradation of the cytoplasma, of intercellular inclusion bodies, strangulation of certain areas of it may play a role. It is assumed that the development of inclusion bodies is a result of impairment of the cell membrane of proliferating, undifferentiated epithelial cells of the colonic mucosa. Increasing frequency of inclusion bodies in polyps undergoing malignant transformation seems to be an important morphologic feature. PMID- 6275263 TI - [Hashish]. PMID- 6275264 TI - Absence of detectable alteration in the kinetoplast DNA of a Trypanosoma brucei clone following loss of ability to infect the insect vector (Glossina morsitans). AB - A monomorphic bloodstream population of Trypanosoma brucei EATRO 1244 was derived from a cloned pleomorphic parental population by 77 rapid passages through mice. Loss of pleomorphism was accompanied by increased virulence of trypanosomes towards the mammal, by loss of ability to infect the tsetse fly, Glossina morsitans, loss of ability to transform to the procyclic stage in vitro at 26 degrees C, and by loss of oligomycin-sensitive ATPase activity in trypanosome homogenates. No differences in the maxicircle component of the kinetoplast DNAs (kDNA) of the two populations were detected by electron microscopy of kDNA network spreads or by electrophoretic analysis of restriction endonuclease digests. It appears, therefore, that loss of transmissibility and associated ability of the trypanosomes to activate the mitochondrion need not necessarily be the result of deletions in the mitochondrial (maxicircle) genome. We suggest that point mutations in critical mitochondrial genes, undetectable using out methods, or mutations of nuclear genes coding for important mitochondrial enzymes, may account for the observed changes in phenotype. PMID- 6275265 TI - ACTH levels after the dexamethasone suppression test in depression. PMID- 6275266 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 5-1982. A 15-year-old boy with slowly progressive dementia. PMID- 6275267 TI - Resistance to antiviral drugs of herpes simplex virus isolated from a patient treated with acyclovir. PMID- 6275268 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 6-1982. A 55-year-old man with eight months of obstructive jaundice. PMID- 6275269 TI - The benzodiazepine receptor: anatomical aspects. PMID- 6275270 TI - Benzodizepines: biochemistry of function. PMID- 6275271 TI - Inward current channels activated by intracellular Ca in cultured cardiac cells. PMID- 6275272 TI - Effects of nifedipine on electrical and mechanical responses of rat and guinea pig vas deferens. PMID- 6275273 TI - Agonist and antagonist benzodiazepine receptor interaction in vitro. PMID- 6275274 TI - Type C virus particles in a cord T-cell line derived by co-cultivating normal human cord leukocytes and human leukaemic T cells. PMID- 6275275 TI - Antagonistic properties of quipazine at presynaptic serotonin receptors and alpha adrenoceptors in rat brain cortex slices. AB - Rat brain cortex slices preincubated with 3H-serotonin or 3H-noradrenaline were superfused with physiological salt solution, and the effect of quipazine on the 3H overflow evoked by electrical field stimulation was examined. 1. The electrically evoked tritium overflow from brain slices preloaded with 3H serotonin was increased by quipazine in a concentration-dependent manner. The concentration-response curves of unlabelled serotonin and noradrenaline for their inhibitory effects on impulse-evoked 3H overflow were shifted to the right by quipazine. 2. In brain slices preincubated with tritiated noradrenaline, quipazine caused an increase in electrically stimulated 3H overflow which was less marked than the effect on brain slices preloaded with 3H-serotonin. The concentration-response curve of unlabelled noradrenaline for its decreasing effect on stimulation-evoked 3H overflow was shifted to the right by quipazine. We conclude that quipazine blocks presynaptic inhibitory serotonin receptors and alpha-adrenoceptors on monoaminergic neurones of the rat brain cortex. PMID- 6275276 TI - Influence of acid-base alterations on myocardial sensitivity to catecholamines. AB - The influence of "respiratory" and "metabolic" acid-base alterations on the myocardial sensitivity to catecholamines was studied in the isolated rat atria. The ability of noradrenaline for increasing the atrial rate was enhanced during alkalosis and conversely, it was decreased by acidosis. These changes in sensitivity shifted the concentration-effect curve for noradrenaline to the right by about 0.5 log unit when the pH was lowered from 7.60 to 7.00. No changes in the maximum attainable response were detected. Essentially the same shifts of the concentration-effect curves were obtained with changes in pH brought about by altering the pCO2 or at constant pCO2. The decrease in the pH produced a similar shift to the right of the concentration-effect curve for isoprenaline, after the extraneuronal uptake inhibition by hydrocortisone and also in atria tissue with low content of endogenous noradrenaline (reserpine-pretreated and newborn rats). The ability of isoprenaline for increasing cyclic AMP levels in atrial tissue was also enhanced by alkalosis and decreased by acidosis. However, the shift to the right of the concentration-effect curve for cyclic AMP induced by the decrease in the pH was greater than the shift detected in the chronotropic-effect curve. In addition a decrease in the maximum increment of cyclic AMP was detected under acidosis, in spite of equal maximal chronotropic response. Our results support the hypothesis that the alterations in the sensitivity to catecholamines induced by the changes in pH are not due to a release of endogenous noradrenaline nor to alterations of the mechanisms which remove catecholamines from the biophase. The fact that cyclic AMP response to catecholamines was also reduced by acidosis strongly suggests that the mechanism(s) involved is located in the earlier steps of the events leading to the chronotropic effect of the beta-agonists. PMID- 6275277 TI - AQA 39*, a new bradycardic agent which blocks myocardial calcium (Ca) channels in a frequency- and voltage-dependent manner. PMID- 6275278 TI - The electrophysiological basis of the bradycardic action of AQA 39 on the sinoatrial node. AB - The electrophysiological effects of the bradycardic agent AQA 39 (5,6-dimethoxy-2 [3-[[ alpha-(3,4-dimethoxy) phenylethyl] methylamino] propyl] phthalimidine hydrochloride) on small preparations of the S-A node of the rabbit heart were investigated. The following results were obtained: 1. The decrease in the rate of the spontaneous preparation resulted from a lower rate of diastolic depolarization, a slower upstroke and a longer duration of the action potential. Concomitantly, overshoot and maximum diastolic potential were decreased. 2. The drug effect on rate strongly depended on the potential during diastole. AQA 39 acted stronger the more positive the maximum diastolic potential. 3. In voltage clamp experiments, the membrane potential was held at -40 mV and transiently depolarized by square pulses to -10 mV. At a low pulsing rate (0.005 Hz), the main effect was a reduction of the time-dependent potassium current (Ik); the slow inward current (Isi) was only slightly reduced. However, when the pulsing rate was increased to 1 Hz, a clear reduction of Isi was observed. 4. When the block of Ca channels had reached a steady state during continuous pulsing in the presence of the drug, part of the block could be removed by rest periods, relief of block being dependent on the membrane potential during rest. At a fixed rest period of 45s, relief of block was nearly complete for potentials negative to -55 mV but negligible positive to -35mV. 5. AQA 39 shifted the dose-response curve to ionophoretic application of acetylcholine to higher concentrations, suggesting a competitive action of the drug with acetylcholine at the muscarinic receptor. PMID- 6275279 TI - Effects of a new antiarrhythmic compound [2-benzal-1-(2' diisopropyl-amino-ethoxy imino)-cycloheptane hydrogen fumarate] on the electrophysiological properties of mammalian cardiac cells. AB - Intracellular microelectrodes were used to study the effects of Th 494 [2-benzal 1-(2' diisopropyl-amino-ethoxy-imino)-cycloheptane hydrogen fumarate; 1-100 mumol/1) on transmembrane electrical activity of sinus node and Purkinje fibres of the rabbit as well as on atrial trabeculae and papillary muscles of the guinea pig. In the atrial and in the ventricular myocardium (32 degrees C; driving rate 0.3-0.5 Hz) Th 494 exerted the following electrophysiological actions: no change of the resting potential nor of the amplitude of the action potential; concentration- dependent reduction of the maximum rate of rise (dV/dt)max of the action potential; slight increase of the action potential duration at lower concentrations (1-20 mumol/l), loss of the plateau at higher concentrations (above 20 mumol/l). The isometric force of contraction was moderately reduced by Th 494 (about 20% reduction by 2 mumol/l). The h infinity-curve relating (dV/dt)max of the action potential to the membrane potential was depressed by Th 494 without being shifted along the voltage axis. The reduction of (dV/dt)max was considerably more pronounced at higher driving frequencies. After interruption of stimulation for various periods, (dV/dt) max of the first action potential attained a steady-state value in a two-exponential fashion, suggesting use dependence as well as a change in the recovery kinetics of the fast Na+ channel by Th 494. In Purkinje fibres (37 degrees C) Th 494 reduced (dV/dt) max in a similar manner. The duration of the action potential was considerably decreased at the level of the plateau. In the primary pacemaker region of the sinus node (37 degrees C) Th 494 moderately reduced the rate of diastolic depolarization and diminished at higher concentrations the amplitude of the action potential. All effects of Th 494 were only slowly reversible by drug-free perfusion. In view of its effect on (dV/dt) max, Th 494 resembles quinidine in its potential dependence, whereas its time-dependence bears greater similarity with lidocaine. PMID- 6275280 TI - Gaba induced excitatory responses in the guinea-pig small intestine are antagonized by bicuculline, picrotoxinin and chloride ion blockers. AB - Contractions of the guinea-pig ileum elicited by GABA were reversibly antagonised by various concentrations of picrotoxinin and bicuculline. This antagonism was specific for GABA-induced responses, responses to ACh, BK, and HA being unaffected. The chloride ion channel blockers, furosemide, 2 x 10-4 mol/l, and piretanide, 5.5 x 10-5 mol/l, substantially reduced GABA-induced contractions of the ileum but not those elicited by ACh or electrical stimulation of the cholinergic nerves. The action of GABA in stimulating the intrinsic cholinergic nerves appears to be Cl- dependent. PMID- 6275281 TI - Further characterization of opioid receptors in the striatum mediating muscular rigidity in rats. AB - In previous studies, we had found that opioid receptors in the striatum can mediate muscular rigidity which can be recorded as a tonic EMG activity in the gastrocnemius-soleus muscle of rats. We have now tried to evaluate the type of opioid receptors mediating the EMG activity. For this purpose, opioids regarded to be selective agonists at one type of opioid receptors were injected into the striatum of unanesthesized rats. Morphine, a micro-type agonist, in the dose of 40 nmol induced a pronounced EMG activity, while 10 to 40 nmol of [D-Ala2, D Leu5] enkephalin, a delta-type agonist, led to no or little effect, respectively. beta-Endorphin, which reacts with the epsilon -type of receptors, was completely ineffective in doses of 3 or 15 nmol. The benzomorphan compound Mr 2033-Cl, which is a kappa-type agonist, induced a moderate activity in the dose of 15 nmol; 3 nmol were ineffective. beta-Casomorphin-4 was the most potent of the drugs studied in our system, since 9 nmol induced a pronounced rigidity, which was naloxone-reversible (2 mg/kg i.p.). The doses of these opioids effective in comparison with their known in vitro potencies suggest that the rigidity is mediated by a group of striatal opioid receptors, which has some similarity to the micro-type, but also shows some differences. In contrast, haloperidol, injected either into the striatum (30 nmol) or systemically (2 mg/kg i.p.), did not induce any EMG activity, suggesting that a decrease in dopaminergic neurotransmission is not the primary mechanism inducing the rigidity. beta-LPH62 77 and beta-LPH66-77 (15 nmol) intrastritally] were also completely inactive. PMID- 6275283 TI - Androgen-associated hepatoma in a hemodialysis patient. AB - A patient on hemodialysis treatment developed hepatocellular carcinoma (HCC) after long-term therapy with androgenic anabolic steroids. The tumor progressed very rapidly, and there was no evidence of regression despite discontinuation of the drug. In view of the evidence of an increased of malignancy in patients with chronic uremia and hemodialysis, and of the higher frequency of HCC correlated to treatment with C17-alkylated anabolic steroids, it is necessary to further evaluate the efficiency of this treatment in the aforementioned group of patients. PMID- 6275284 TI - Effect of renal artery occlusion on plasma cAMP. PMID- 6275282 TI - Resistance of adrenergic neurotransmission in the toad heart to adrenoceptor blockade. AB - Stimulation of sympathetic nerves to the toad heart produced increases in both the rate and force of cardiac beat. Although these responses were abolished by treatment with bretylium (10-6 mol.1-1) or 6-hydroxydopamine (100 mg.kg-1), and surgical sympathetic denervation, they were not abolished by treatment with propranolol (10-6 mol-1-1) and phentolamine (3 X 10-6 mol.1-1), either alone or in combination. The responses remaining after adrenoceptor blockade could not be ascribed to the effects of neurally released dopamine, ATP, adenosine, histamine or a variety of neuropeptides, although the participation of an as yet unidentified co-transmitter cannot be ruled out. Quantitative analysis of the interactions between propranolol and adrenaline on cardiac adrenoceptors, after blockade of alpha-receptors and amine uptake mechanisms, revealed that these interactions do not comply with the conditions for simple competitivity. Therefore, in addition to its action on beta-receptors, adrenaline seems to be producing excitation of the toad heart by acting on adrenoceptors which cannot be classified as either alpha- or beta-receptors. These results, together with the existence of close neuromuscular gaps (less than 50 nm) in the toad heart, are consistent with the hypothesis that sympathetic excitation of the toad heart is mediated by both "extra-junctional" beta-adrenoceptors, and "junctional" adrenoceptors which are neither alpha- nor beta-receptors. PMID- 6275285 TI - Levels of dynorphin-(1-13) immunoreactivity in rat neurointermediate pituitaries are concomitantly altered with those of leucine enkephalin and vasopressin in response to various endocrine manipulations. AB - The levels of dynorphin-(1-13), leucine enkephalin, beta-endorphin and vasopressin immunoreactivity (ir-DYN, ir-1-ENK, ir-beta-END, ir-VP) have been determined in the anterior and in the neurointermediate lobes of the pituitary of rats subjected to a variety of manipulations. Dehydration of rats by 5 days enforced inhibition of a 2% solution of NaCl resulted in a significant decrease in the levels of ir-DYN, ir-1-ENK and ir-VP, but not in those of ir-beta-END in the neurointermediate lobe of the pituitary. In contrast, substitution of drinking water by a solution containing 20 microgram/ml dexamethasone for 5 days produced a significant increase in the neurointermediate pituitary content of ir DYN, ir-1-ENK and ir-VP, whereas levels of ir-beta-END remained unaffected. This treatment, however, resulted in a significant fall in the ir-beta-END content of the adenopituitary without changing levels of ir-DYN in this structure. Adrenalectomy was associated with a significant decrease in the ir-DYN, ir-VP and ir-1-ENK content of the neurointermediate lobe of the pituitary and a pronounced elevation in the ir-beta-END but not ir-DYN content of the adenohypophysis. These observations are indicative that the regulation mechanisms of the functional state of particular endorphins differ between the anterior and neurointermediate lobes of the pituitary. The concomitant alterations in levels of ir-DYN, ir-1-ENK and ir-VP detected suggest that a common or similar mechanism of regulation may exist for these peptides. A common biosynthetic origin, however, appears to be unlikely, since Brattleboro rats which are unable to synthesize vasopressin possess unchanged ir-DYN- and ir-1-ENK- levels in the pituitary. PMID- 6275286 TI - Temporal and other effects of catechol estrogens on prolactin secretion in the rat. AB - Temporal alterations in plasma prolactin levels caused by the administration of 2 hydroxyestradiol and 2-hydroxyestrone (100 microgram/kg) into the right atrium of freely-moving conscious male rats were examined. The catechol estrogens were given in a single bolus via an indwelling cannula and plasma prolactin concentration was monitored by taking blood samples every 2 min. A pulsatile elevation of plasma prolactin occurred approximately 4 h after the injection of 2 hydroxyestradiol and a small increase was also observed when it was administered to rats bearing a Silastic capsule containing estradiol. 2-Hydroxyestrone had no effect in untreated male rats but produced a 5- to 6-fold elevation in plasma prolactin level 4 h after its administration to rats implanted with estradiol. It is proposed that 2-hydroxyestrone suppresses the action of estradiol on prolactin secretion from the pituitary and that the accumulated hormone is released when the concentration of this catechol estrogen falls below a critical level. A longer latent period was required to produce an elevation in plasma prolactin levels by 2-hydroxyestradiol than by estradiol. PMID- 6275287 TI - Tolerance and dependence of dorsal horn neurones of the cat: the role of the opiate receptors of the substantia gelatinosa. PMID- 6275288 TI - Evidence for central selective dopamine receptor stimulation in the mediation of nomifensine-induced hyperalgesia and the effects of opiate antagonists. PMID- 6275289 TI - Binding of (3H)-5,5 diphenylhydantoin to rat brain membranes. PMID- 6275290 TI - Cyclic GMP in rat hippocampus: regulation by the septo-hippocampal pathway. PMID- 6275291 TI - Phospholipids and benzodiazepine recognition sites of brain synaptic membranes. PMID- 6275292 TI - Multiple opiate receptors: differential binding of mu, kappa and delta agonists. PMID- 6275293 TI - Chronic ethanol imbibition interferes with delta-, but not with mu-opiate receptors. PMID- 6275294 TI - Ionic currents in the somatic membrane of rat dorsal root ganglion neurons-I. Sodium currents. PMID- 6275295 TI - Ionic currents in the somatic membrane of rat dorsal root ganglion neurons-II. Calcium currents. PMID- 6275296 TI - Ionic currents in the somatic membrane of rat dorsal root ganglion neurons-III. Potassium currents. PMID- 6275297 TI - Comparative toxicity of kainic acid and other acidic amino acids toward rat hippocampal neurons. PMID- 6275298 TI - Spontaneous and potassium-induced release of acetylcholine from mouse forebrain minces. PMID- 6275299 TI - A receptor for protons in the membrane of sensory neurons may participate in nociception. PMID- 6275300 TI - Electrophysiological demonstration of a dentato-rubrospinal projection in cats. PMID- 6275301 TI - Changes in histamine in the rat striatum following local injection of kainic acid. PMID- 6275302 TI - Ionic mechanisms of alpha-autoinhibition of [3H]noradrenaline secretion in guinea pig vas deferens: possible role of extracellular sodium. PMID- 6275303 TI - Prostaglandins in denervated skeletal muscle of the rat: effect of direct electrical stimulation. PMID- 6275304 TI - Neuromuscular transmission in neonatal mice injected with serum globulin of myasthenia gravis patients. AB - Neuromuscular transmission was studied in neonatal mice following injection with serum globulin of patients with myasthenia gravis (MG). Compared to controls, these mice showed significant reduction in successive muscle action potentials evoked by repetitive nerve stimulation, amplitude of miniature endplate potentials, and postjunctional sensitivity to acetylcholine. There was no change in evoked isometric tension, quantal content of endplate potentials, or input resistance of the endplate membrane. These results confirm earlier reports of neuromuscular block in animals following injection of globulin of myasthenic patients, and demonstrate that decrease in amplitude of evoked potentials and of miniature endplate potentials is due to reduction in sensitivity to acetylcholine rather than in input resistance of the postsynaptic membrane. These findings are compatible with a postsynaptic defect in MG caused by a humorally mediated autoimmune mechanism. PMID- 6275305 TI - gamma-Aminobutyric acid receptors in normal human brain and Huntington disease. AB - GABA receptor binding curves were measured in well-washed membrane homogenates from eleven regions of normal human brain and four regions from brains of Huntington disease patients. Computer analysis suggested two populations of receptor sites of different affinity (KD = 10 nM and 240 nM) present in all brain regions but in variable quantity (cerebellar cortex and cerebral cortex greater than basal ganglia greater than deeper brain structures). The number of GABA receptor sites in the caudate-putamen region of Huntington brain was less than normal, but the number of sites was increased in Huntington disease substantia nigra. No differences from normal were found in cerebellar cortex or frontal cortex from Huntington disease, and no significant changes in binding affinity were observed for any of the four regions tested under the conditions used. PMID- 6275306 TI - Absence of evoked potential abnormalities in acute transverse myelopathy. AB - We studied 12 consecutive patients with inflammatory acute transverse myelopathy (ATM) and no prior history of neurologic disease. All had normal pattern shift visual and brainstem auditory evoked potentials. All of nine patients tested also had normal median nerve short-latency somatosensory evoked potentials. None developed new neurologic signs in 18 months' mean follow-up. In contrast, 72% of 50 patients with definite, probable, or possible multiple sclerosis (MS) had abnormalities in at least one evoked potential test. When defined as a virtually or completely transverse inflammatory lesion, acute transverse myelopathy differs from MS. PMID- 6275307 TI - Phenytoin prolongs far-field somatosensory and auditory evoked potential interpeak latencies. PMID- 6275308 TI - Trigeminal nerve stimulation: short latency somatosensory evoked potentials. AB - We have devised a technique to measure trigeminal somatosensory evoked potentials, stimulating the mentalis nerve at the mandibular foramen and recording the evoked early potential on the scalp at 3.2 msec. The early potential was easily recognized in all normal subjects and on the asymptomatic side of patients with facial pain. The latency of the early potential in normals was 1 to 2 msec less than half the latency of the mechanically induced jaw jerk. The early potential appeared simultaneously on both sides after stimulation of either the right or left mentalis nerve and may have its origin in deep brainstem structures, the Gasserian ganglion, or the trigeminal nerve. The technique is useful in documenting the functional integrity of peripheral and central afferent pathways of the trigeminal nerve. PMID- 6275309 TI - Lymphocyte abnormality in human myotonic dystrophy and experimental drug-induced myotonia. AB - In the cytoplasm of peripheral blood lymphocytes in 3 of 13 patients with myotonic dystrophy, myelin-like structures were observed electronmicroscopically. Some were connected to the cytoplasmic membranes, and some were surrounded by a limiting membrane possessing acid phosphatase activity. These findings suggest an aberration of cytoplasmic membranes in lymphocytes. Similar structures had appeared in lymphocytes of normal rats that became myotonic with 20, 25 diazacholesterol. These findings suggest that a primary genetic defect in human myotonic dystrophy, which participates in the formation of myelin-like structures in lymphocytes, might be also responsible for the occurrence of a myotonic phenomenon. PMID- 6275310 TI - Epstein-Barr in acute nontraumatic mononeuropathies. PMID- 6275311 TI - [Symptomatology and the resectability of bronchopulmonary carcinoma]. PMID- 6275312 TI - Retinal projection to the nucleus of the optic tract in the cat as revealed by retrograde transport of horseradish peroxidase. AB - Retrograde transport of horseradish peroxidase injected iontophoretically into the nucleus of the optic tract of cats revealed that the direction-selective cells in this pretectal nucleus receive direct retinal projections from small retinal ganglion cells, the so-called gamma-cells. These cells from a horizontal band on the contralateral retina. Few labeled cells are found in the ipsilateral temporal retina. The input from the contralateral retina is 10 times more numerous than from the ipsilateral one. In both retinae the highest concentration of labeled cells is near the area centralis. PMID- 6275313 TI - Origins of the pretectal and tectal projections to the central lateral nucleus in the cat. AB - The pretectal and tectal projections to the thalamic intralaminar nuclei in the cat were studied following horseradish peroxidase injections centered in the central lateral nucleus. Retrogradely labeled neurons were found in both the pretectum and the superior colliculus, ipsilateral and, to a lesser extent, controlateral to the injection site. Labeled pretectal neurons were found throughout all pretectal nuclei; the densest concentrations were in the medial pretectal nucleus, the anterior pretectal nucleus and the nucleus of the posterior commissure. The major source of tectal projections were the small neurons in the stratum griseum intermediate. PMID- 6275314 TI - Neuronal and glial cells in the superficial layers of early postnatal mouse neocortex: immunofluorescence observations. AB - Sections of immature (postnatal day 5) mouse cerebral cortex was examined for several cell-type specific immunological markers. Glial fibrillary acidic (GFA) protein or vimentin were detected in astrocytic cell processes and--more rarely- cell bodies located in the superficial layers, but not within putative Cajal Retzius cells (CRs). These cells did, however, react with cholera toxin, tetanus toxin and NS-4 antibodies. In agreement with previous ultrastructural observations, we conclude that CRs are neurons, or at least cells which display the basic characteristics of neurons. PMID- 6275315 TI - Voltage-clamp studies of a slow inward current in bullfrog sympathetic ganglion cells. AB - Voltage-dependent inward membrane currents of bullfrog sympathetic ganglion cells were analyzed with the voltage clamp method. Two distinct inward currents, a tetrodotoxin (TTX)-sensitive fast inward current (INa) and a TTX-insensitive slow inward current (Isi), were recorded in Ringer solution. The Isi was markedly depressed by removal of external Ca2+ and by the addition of Mn2+, Co2+ or D-600. This suggests that the Isi is, almost exclusively, due to Ca2+. These results indicate that under physiological conditions a Ca2+ influx is induced during initiation of action potentials in these ganglion cells. PMID- 6275316 TI - Long-term potentiation investigated in a slice preparation of striate cortex of young kittens. AB - In slice preparations from kitten striate cortex, long-term potentiation (up to 15 h) of field potentials was demonstrated after conditioning stimulation of white matter. Current source-density analysis and measurement of response latencies of cortical units indicated that the potentiation represented enhancement of both geniculo-cortical transmission in layer IV and intracortical transmission in layer II--III. PMID- 6275317 TI - Desensitization-like changes in GABA receptor binding of rat fascia dentata after entorhinal lesion. AB - Destruction of the hippocampal perforant path fibers reduced the binding of [3H]GABA to membranes prepared from the rat fascia dentata. This result could be detected 1--4 months after surgery, but not in 10 days or less. Such a delayed response appears most compatible with a transsynaptic effect on GABA receptors. Values of low affinity KD and Bmax decreased by about 65%, but no changes were detected in high affinity binding, Hill slope or pharmacological specificity. These findings are consistent with a desensitization of postsynaptic low affinity GABA receptors, possible caused by excessive release of GABA. PMID- 6275318 TI - Calcium spike induced by electrical stimulation to the sensory nerve terminal of the frog muscle spindle. AB - A calcium spike in the sensory nerve terminal of the frog muscle spindle could be elicited by electrical pulses, which were given across an air-gap on which the first myelinated segment of the parent axon outside the spindle capsule was bridged, after the sodium spike had been blocked by treatment with tetrodotoxin (TTX). The threshold current was 1.1-2.4 nA higher than that for the sodium spike, suggesting that the calcium channels distribute distally to the site of afferent impulse initiation, or that the density of calcium channels on the encoding site may be less than that along portions distal to the site. PMID- 6275319 TI - Changes in rat hippocampal benzodiazepine receptors and lack of changes in muscarinic receptors after fimbria-fornix lesions. AB - The binding of [3H] L-quinuclidinylbenzilate to muscarinic receptors and [3H] flunitrazepam to benzodiazepine receptors were studied in the rat hippocampus after lesion of the fimbria-fornix. While the muscarinic receptors showed no change, the benzodiazepine receptors did change considerably at various time intervals. Two days after the lesion the specific [3H] flunitrazepam binding decreased 38%, while at 5 days it increased 65%. After 14 days of the lesion it still was significantly above normal. These changes are due to a variation in the number of sites, and not in affinity. Possible interpretations of these results are discussed. PMID- 6275321 TI - Modification of sodium channel kinetics by the insecticide tetramethrin in crayfish giant axons. AB - The effects of the pyrethroid insecticide tetramethrin on voltage-dependent sodium channels were studied with internally perfused crayfish giant axons. At low concentrations in the order of 10-8-10-9M, tetramethrin caused an increase in depolarizing after-potential which in turn triggered repetitive after-discharges. Under Voltage clamp conditions, the sodium current was markedly prolonged during a step depolarization, and a large and prolonged sodium tail current appeared upon step repolarization. A population of sodium channels having activation and inactivation kinetics identical to those in control axons was observed in the tetramethrin-poisoned axons, indicating that only a fraction of the channels was modified. The modified channels exhibited remarkably slow kinetics, activating with a time course of 100 msec to 2 sec and inactivating with a time course of 1 5 sec depending on the membrane potential. The voltage dependence of the modified channels was shifted in the direction of hyperpolarization by about 10-20 mV with respect to normal sodium channels. The large inward sodium tail current associated with step repolarization of the membrane decayed with a time course of 20-600 msec. A kinetic hypothesis describing the behavior of sodium channels in a tetramethrin-poisoned axon is presented and discussed in relation of the behavior of the sodium channels modified by other toxins. PMID- 6275322 TI - Syringocystadenoma papilliferum of the eyelid. AB - Two patients presented with slowly developing papillated or hyperkeratotic lesions of their lid margins that began early in the fourth decade. Upon excision, these tumors were discovered to be examples of syringocystadenoma papilliferum, a lesion not previously well documented in the ophthalmic literature. By light microscopy, each tumor displayed surface poral openings where the epidermis underwent a transition into duct-like epithelium, which ramified as luminal channels throughout the tumor. Papillae projected into many of the duct-like spaces and were lined by columnar to multilaminar nonkeratinizing epithelium. A plasma cell-rich infiltrate was present in the connective tissue cores of the papillae. Electron microscopy performed on one of the lesions revealed that the channels were true ducts, with the innermost cells displaying microvilli, small numbers of tonofilaments, and apical junctional complexes composed of a zonula occludens and zonular adherens. These findings, coupled with the location of the tumors at the lid margin and the presence of dilated apocrine glands of Moll deep to one of the tumors, support an apocrine origin. The clinical differential diagnosis of syringocystadenoma papilliferum with respect to other more common lid tumors is discussed. PMID- 6275320 TI - Differentiation of agonist conformation and antagonist conformation in multiple opioid receptors. AB - To differentiate the opiate (naloxone) receptor and the enkephalin receptor in rat brain, we solubilized the receptor molecules by detergent and determined the molecular weights by gel filtration. The receptor preparation was bound to [3H] naloxone or [3H] Met5-enkephalin, and was solubilized by Triton X-100. On gel chromatography with a Sepharose 6B column, the agonist and the antagonist conformation of opioid receptors eluted as molecules with the molecular weights of 240,000, and 120,000 and with Stokes' radii of 5.5 nm and 4.3 nm, respectively. Further, it was also disclosed that Na+ was bound to the antagonist conformation of opioid receptors but not to the agonist conformation. PMID- 6275323 TI - Eyelid fibrous histiocytoma. PMID- 6275324 TI - Fibrous histiocytoma of the corneoscleral limbus: The ultrastructure of a distinctive inclusion. AB - A case of epibulbar fibrous histiocytoma is presented, with emphasis on the electron microscopic aspects due to the paucity of ultrastructural descriptions of this rare tumor. A tan-yellow conjunctival tumor developed at the corneoscleral limbus in a 31-year-old male. The clinical diagnosis was confirmed by routine histopathologic evaluation, which demonstrated a typical matted or storiform cellular pattern. Electron microscopy revealed a mixed cell population, composed primarily of histiocytic and fibroblastic cells. The histiocytic cells contained lysosomes and phagosomes and had interdigitating cell processes. The fibroblastic cells were characterized by prominent rough-surfaced endoplasmic reticulum and variable amounts of lipid within vacuoles, but did not possess lysosomes or phagosomes. Both cells occasionally contained an unusual curvilinear structure, suggesting a shared histogenesis. This distinctive inclusion appeared to be a modification of the rough-surfaced endoplasmic reticulum. The nature and origin of the tumor cells are discussed on the basis of their ultrastructure. PMID- 6275325 TI - Proton-beam irradiated epithelioid cell melanoma of the ciliary body. AB - A malignant ciliary body melanoma received proton-beam irradiation. After an apparent failure of the tumor to respond, the eye was enucleated. A predominantly epithelioid cell tumor appeared viable by light microscopy, and a low degree of mitotic activity persisted, despite therapy. The tumor cells, however, displayed degenerative changes ultrastructurally, presumably results of the radiotherapy. These consisted of numerous cytoskeletal filaments, lipid vacuoles, prominent phagolysosomes, and nuclear convolutions and fragmentations. The mitochondria were fewer in number in the present tumor than typically encountered in epithelioid cells. A rare leptomeric structure was discovered, probably an organizational modification of the cytoplasmic filaments. The tumor's capillaries showed radiation-induced changes in terms of thickened basement membranes and perivascular fibrin deposition. The foregoing features are indicative of cellular and metabolic injury from the radiotherapy, but these were evidently not sufficiently injurious to sterilize the tumor. PMID- 6275326 TI - Electron microscopy for the diagnosis of ocular viral infections. AB - Tear samples obtained from patients with suspected viral conjunctivitis were examined by virus culture and by the pseudoreplica method for electron microscopy (EM). Adenoviruses were isolated in 13 of the 34 cases (38%) clinically diagnosed as adenovirus conjunctivitis. Characteristic adenoviral particles were demonstrated by electron microscopy in seven of the culture-positive, and two of the culture-negative samples. In cases of herpes simplex virus ocular infection, tears were virus positive by culture in one-third of the patients; two-thirds of the culture positive specimens were also positive by EM. This study demonstrates the potential of electron microscopy as a rapid, specific, and relatively inexpensive method for the laboratory diagnosis of ocular viral infections. PMID- 6275327 TI - Early assessment for discharge planning. PMID- 6275328 TI - Nursing responsibilites for discharge planning in a community hospital. PMID- 6275329 TI - [Pregnancy and placental proteins (SP1, HCG, PP5, alpha 2-PAG) in trophoblastic tumor diseases]. PMID- 6275330 TI - Dysentery form gastroenteritis in infancy. PMID- 6275331 TI - Mandibular and para-mandibular tumors in children. Report of 16 cases. AB - Sixteen cases of mandibular tumors or paramandibular soft tissue tumors with mandibular involvement are reported. These include such rare mandibular tumors or tumor-like conditions as melanotic progonoma, intraosseous haematoma secondary to von Willebrand's disease, post-irradiation osteosarcoma, monostotic eosinophilic granuloma, aneurysmal bone cyst and osseous hemangiopericytoma. Three cases of cherubism, one of fibrous dysplasia or aggressive fibromatosis and one of central giant cell reparative granuloma are also reported. The soft tissue tumors comprise round cell sarcoma, parotid adeno-carcinoma with generalised metastases, embryonal rhabdo-myo-sarcoma, neuro-fibro-sarcoma and congenital cystic hygroma. In all the cases the disease was well advanced when the patient presented for X ray examination. The specific X-ray diagnosis of mandibular and paramandibular tumors in childhood is more difficult than that of similar tumors in other parts of the body. PMID- 6275332 TI - Granular cell myoblastoma of the trachea. PMID- 6275333 TI - Gallium-67 whole body scintigraphy in cytomegalovirus (CMV) mononucleosis. PMID- 6275334 TI - Effect of cortisone on postnatal development of ion transport in rabbit small intestine. AB - To study the effect of corticosteroids on postnatal maturation of Na+ transport in the small intestine, we studied 10-12-day-old suckling rabbits after they had received cortisone acetate, 20 mg/kg SC on days 3, 4, and 5 of life. When killed, the cortisone-injected animals weighed significantly less than saline-injected controls. In jejunal villus enterocytes isolated from this cortisone-treated group, the specific activities of sucrase and Na+-K+-ATPase were significantly greater than those in control enterocytes. Studied in Using chambers, a significant electrical and ion-flux response to glucose was observed in the jejunal epithelium of the treated group, but not in controls. We conclude that exogenous cortisone, administered early in life, can stimulate the precocious development not only of certain epithelial enzymes but also glucose-facilitated Na+ transport in the jejunum of the rabbit. PMID- 6275336 TI - Plant fiber intake in the pediatric diet. PMID- 6275335 TI - Lung surfactant in an era of replacement therapy. AB - In this paper current knowledge of the pulmonary surfactant system with particular emphasis on aspects that relate to exogenous surfactant replacement therapy for the neonatal respiratory distress syndrome (RDS) is considered. The work provides an overview, but concentrates on several facets of lung surfactant research that help to elucidate and evaluate past and present attempts toward such therapy. Subjects addressed include the functional need for specific lung surfactant components to obtain optimal surface activity and some of the required surface property measurements to characterize such activity. Also discussed is current knowledge of lung surfactant synthesis and secretion and the potential effect of exogenous surfactant on the endogenous pulmonary surfactant system. A primary theme throughout is that an analysis of previous clinical trials involving surfactant replacement shows the necessity for strong interactions with related basic science investigations. It is suggested that future human infant surfactant replacement trials proceed deliberately, and include maximal correlations with basic in vitro research on lung surfactant biophysics and biochemistry and with experiments in animal models. PMID- 6275337 TI - [Various problems of the etiology of serous meningitis in children]. PMID- 6275338 TI - The Epstein-Barr virus: historical, biologic, pathologic and oncologic considerations. PMID- 6275339 TI - Prenatal cytomegalovirus infection: epidemiology, pathology and pathogenesis. PMID- 6275340 TI - Effects of uridine triphosphate on contractility, cyclic nucleotide levels and membrane potential in the isolated frog ventricle. AB - A study has been made of the effects of uridine 5'-triphosphate (UTP) on the isolated frog ventricle. Preparations were superfused with solutions containing different concentrations of UTP, and changes in contractility, cyclic nucleotide levels and membrane potential were measured. UTP produced a long lasting increase in isometric twitch tension, which was unaffected by adrenergic receptor antagonists (propranolol and phentolamine). The levels of adenosine 3',5'-cyclic monophosphate (cyclic AMP) and guanosine 3',5'-cyclic monophosphate (cyclic GMP) were measured at different times during exposure of the ventricle to 10(-4) M UTP. The increase in the force of contraction was found to be accompanied by a rise in intracellular cyclic AMP. Cyclic GMP levels were seen to fall initially, but then to increase later, as both twitch and cyclic AMP started to decline. UTP also produced marked changes in the shape of the action potential; its duration and positive overshoot were both increased. The effects of UTP on twitch tension, cyclic nucleotide levels and action potential parameters were all dose-related. The change in contractility was found to be linearly related to (a) the ratio of the amount of cyclic AMP: cyclic GMP present in the fibres, and (b) to the increase in duration of the action potential. These results suggest that cyclic AMP, cyclic GMP and the availability of Ca2+ may all be involved in mediating the response to UTP. PMID- 6275341 TI - Tubular adaptation to Pi restriction in hypophysectomized rats. AB - The renal tubule adapts its tubular transport capacity for inorganic phosphate (Pi) in response to a reduction in the Pi supply. In order to assess whether growth hormone plays a critical role in this adaptive response we have studied the change in the tubular handling of Pi which follows Pi restriction in hypophysectomized (HPX) rats and compared it to that occurring in intact counterparts. HPX and intact rats were maintained either on a 1.2 g/100 g phosphorus diet or fed a 0.2 g/100 g phosphorus diet for 3, 6 or 12 days. HPX rats received ACTH and thyroxine in doses which normalize their low glomerular filtration rate (GFR). Then the maximal net Pi reabsorption per volume of glomerular filtrate (max. TRPi/ml GFR) were determined during acute Pi infusion by clearance technique. The results indicate that HPX rats responded to Pi restriction by raising their tubular capacity to reabsorb Pi. However, the rapidity and the magnitude of the adaptive response was significantly less in HPX than in intact rats. The adaptation to Pi restriction was also observed in HPX rats after thyroparathyroidectomy. It is concluded that growth hormone and other pituitary hormones do not play a key role in the adaptive response to Pi restriction. The reduced adaptive response observed in HPX rats with intact thyroparathyroid glands could be due to the decreased Pi demand consecutive to impaired growth. PMID- 6275342 TI - Rate of pyridine nucleotide oxidation and cytochrome oxidase interaction with intracellular oxygen in hearts from rats with compensated volume overload. PMID- 6275344 TI - Analysis of miniature spontaneous inhibitory postsynaptic currents (sIPSCs) from current noise in crayfish opener muscle. AB - Deviating from the normal situation, some crayfish muscle fibres showed spontaneous inhibitory activity: discharge of large inhibitory postsynaptic currents, IPSCs, alternating with long lasting bursts of current noise. Analysis of the bursts of current noise revealed that they are composed of spontaneous miniature unit currents, sIPSCs. In the burst periods the sIPSCs occurred with an average rate of 3.5--10 k Hz and had an amplitude of about alpha = 90 pA at a driving force delta E = 10 mV. The peak conductance gamma alpha = alpha/delta E of the sIPSCs was gamma alpha = 9.2 nS +/- 0.5 (S.D., n = 5) for membrane potentials between E = --60 mV and E = --80 mV. gamma alpha seemed to decrease when the membrane was hyperpolarized. The time constants of decay, tau of the sIPSCs were identical with tau of the IPSCs. Further, tau and its potential dependence agreed with the mean lifetimes of inhibitory postsynaptic channels operated by gamma-aminobutyric acid (GABA) [Dudel et al. 1977, 1980]. Synchronized opening of about 750 inhibitory synaptic channels generates a sIPSC. Analysis of this anomalous bursting inhibitory activity thus yields the size of the inhibitory quantum of transmission, which could not be obtained from IPSCs. PMID- 6275343 TI - Effect of triiodothyronine on (Na+ G K+)-adenosine triphosphatase and (Na+ + Mg2+)-dependent phosphorylated intermediate in rat salivary glands. AB - The present studies concern the effect of triiodothyronine (T3) on rat salivary (Na+ + K+)-adenosine triphosphatase (NaK-ATPase). The results indicate that T3 selectively increased submandibular and parotid NaK-ATPase units with a single, large dose of T3, different dosage of T3, three successive doses of T3 given on alternate days, and "physiological" doses of T3 with daily injection for 2 weeks. Sublingual NaK-ATPase was insensitive to T3 in the above experiments. The effect of T3 on submandibular and parotid NaK-ATPase is selective since Mg-ATPase and 5' nucleotidase showed no significant differences under different thyroid status. The lack of response of NaK-ATPase to reverse T3, further substantiates the specificity of T3 on salivary NaK-ATPase. PMID- 6275346 TI - Feed composition and left abomasal displacement in dairy cattle. A case-control study. AB - The present case-control study elucidates the significance of the crude fiber content in the ration for the development of left abomasal displacement. On the basis of approximate relative risk estimations a crude fiber content in the complete ration higher than 16 to 17% is recommended for the prevention of abomasal displacement. PMID- 6275345 TI - Growth of sympathetic postganglionic axons after partial chemical destruction with guanethidine. AB - Partial sympathectomy of neonatal rats was produced by treatment with guanethidine. The number of neurons of the superior cervical ganglion decreased to 15% of control values and remained constant throughout the subsequent observation period of 4 months. The volume of the remaining neuronal perikarya increased faster than that of control animals during the observation period, and the density of adrenergic innervation of the iris and the noradrenaline content of the heart were found to partially recover after the initial decrease produced by the guanethidine treatment, reaching 33% and 30% of control values, respectively, after 4 months. The noradrenaline content of the duct of the vas deferens was greatly reduced by guanethidine treatment, but almost recovered after 4 months. The frequency of sympathetic preganglionic impulses was elevated throughout the 4 months observation period, and it is suggested that this increase is responsible for the compensatory changes in cell volume, terminal density and terminal noradrenaline content of the remaining neurons. PMID- 6275347 TI - [Cauda equina syndrome complicating enkylosing spondylarthritis]. PMID- 6275348 TI - [Ectopic pheochromocytoma in black Africans : rule or exception? Apropos of a case of recurrent ectopic pheochromocytoma]. PMID- 6275350 TI - [Cure of herpetic encephalitis with adenine arabinoside]. PMID- 6275349 TI - [Location of an insulinoma by transhepatic phlebography with staged insulin assays (author's transl)]. AB - Insulinoma was easily diagnosed in a 73-year-old woman who had organic hypoglycaemia associated with hyperinsulinism, but the tumour could not be located by echotomography and computerized tomography. The state of the patient's arteries precluded arteriography. Pancreatic phlebography was carried out by the portal transhepatic route and blood was collected at different levels for plasma insulin assays. A very high gradient at the pancreatic isthmus indicated the site of the tumour, which was found on surgery to be precisely there and could be enucleated. This technique cannot be used systematically to locate insulinomas, but it is unquestionably helpful when the tumour cannot be located by other methods or when these are contra-indicated. PMID- 6275351 TI - [Mycosis fungoides associated with primary cancer of the liver in an African patient]. PMID- 6275352 TI - Mononucleotide and dinucleotide frequencies, and codon usage in poliovirion RNA. AB - The polio type 1 (Mahoney) RNA sequence (1) has been analyzed in terms of the distribution of its mononucleotides, dinucleotides and trinucleotides (codons). The distribution of adenosine in the sequence is nonuniform, being lower at the 5' end and higher at the 3' end. The dinucleotide CG is relatively rare and the dinucleotides UG and CA are relatively more common than expected. Codon usage is decidedly nonrandom. Codons containing CG are avoided and those ending in adenosine are favored. The asymmetric use of mononucleotides, dinucleotides and codons in polio RNA is unexplained at the present time although the lowered CG frequency may be the result of a DNA origin for polio RNA. PMID- 6275354 TI - A small segment of polyoma virus DNA enhances the expression of a cloned beta globin gene over a distance of 1400 base pairs. AB - The hemoglobin beta-1 gene of the rabbit was linked to a 244 bp DNA fragment from the beginning of the polyoma virus late region, not including the viral origin of replication. After transfection of such recombinant DNAs into mouse 3T6 and human HeLa cells, the polyoma sequences were found to strongly enhance the level of correct beta-globin gene transcripts over a distance of at least 1400 bp. These findings are similar to those obtained with a segment of DNA from the corresponding region of the SV40 genome (J. Banerji, S. Rusconi and W. Schaffner, 1981, Cell, in press) which, however, shows very limited sequence homology to the polyoma 244 bp segment. Using the same assay, a complete copy of polyoma virus DNA was found to interfere with the enhancement of globin gene expression in a cell type-specific manner which may be due to incorrect transcription. In contrast to the complete polyoma virus genome, the 244 bp DNA fragment will be particularly useful as a component of mammalian expression vectors since it almost exclusively yielded high levels of correct beta-globin gene transcripts. PMID- 6275355 TI - Dissection of the r-determinant of the plasmid R100.1: the sequence at the extremities of Tn21. AB - We have sequenced the extremities of the transposon Tn21, isolated from the r determinant of the multiple antibiotic resistance plasmid R100.1, and show that it is a member of the Tn3 family of elements. PMID- 6275353 TI - A region of the polyoma virus genome between the replication origin and late protein coding sequences is required in cis for both early gene expression and viral DNA replication. AB - Deletion mutants within the Py DNA region between the replication origin and the beginning of late protein coding sequences have been constructed and analysed for viability, early gene expression and viral DNA replication. Assay of replicative competence was facilitated by the use of Py transformed mouse cells (COP lines) which express functional large T-protein but contain no free viral DNA. Viable mutants defined three new nonessential regions of the genome. Certain deletions spanning the PvuII site at nt 5130 (67.4 mu) were unable to express early genes and had a cis-acting defect in DNA replication. Other mutants had intermediate phenotypes. Relevance of these results to eucaryotic "enhancer" elements is discussed. PMID- 6275356 TI - The 5' termini of RNAs encoded by the transposable element copia. AB - The 5' termini of copia-specific RNAs in Drosophila melanogaster tissue culture cells were determined by S1 nuclease mapping and cap analysis. Both major copia RNAs share an identical set of heterogeneous 5' ends. Three major cap 1 structures M7GpppCmpUp, M7GpppCmpCp and M7GpppGmpUp together with several other minor caps were found. Almost all the 5' termini, as judged by S-1 nuclease mapping, were located either in a pyrimidine-rich part of the terminal direct repeat or apparently outside of the copia element, suggesting that a proportion of copia transcripts derive from promoters external to the genetic element. PMID- 6275357 TI - Sequences at the capped 5'-ends of polyoma virus late region mRNAs: an example of extreme terminal heterogeneity. AB - We have localized with respect to the genomic DNA sequence the capped 5'-termini of polyoma virus late region mRNAs. A minimum of fifteen different purine termini were found within a 94 base pair region (66.36 to 68.12 map units, nt 5075-5168) immediately preceding the sequence determining the late region mRNA leader repeat (1-3). The most common termini occur at nearly every possible purine within a 25 bp sequence proximal to the leader repeat unit. These do not bear the usual positional relationship to a sequence resembling the 'TATA' box consensus. Deletion mutants lacking minor cap sites and sequences upstream from the principal cap sites were viable. A deletion mutant lacking one of the principal cap sites formed small plaques, while a slightly larger deletion further impinging on the principal cap site region was non-viable. The principal cap sites, which we assume to be transcriptional initiation points, are included in a DNaseI hypersensitive region of polyoma virus chromatin (4). PMID- 6275358 TI - The actin gene in yeast Saccharomyces cerevisiae: 5' and 3' end mapping, flanking and putative regulatory sequences. AB - The 5' and 3' flanking regions of the yeast actin gene have been sequenced and the ends of the actin mRNA were determined by the single-strand nuclease mapping procedure. The mRNA starts with a pyrimidine residue 141 (or 140) nucleotides upstream from the initiation codon. The actin gene lacks a typical "TATA" box 30 base pairs upstream from the mRNA start site but it contains a region homologous to the canonical sequence 5'-GGCTCAATCT-3' which is found in several eukaryotic genes 70 to 80 bp upstream from the mRNA cap site. Judging from the S1 nuclease mapping, there are two populations of actin mRNA terminating 98 and 107 nucleotides downstream from the stop codon. The 3' termini are preceded by three AATAAA sequences found in most eukaryotic polyadenylated mRNAs. PMID- 6275359 TI - Structure and sequence of the mitochondrial 20S rRNA and tRNA tyr gene of Paramecium primaurelia. AB - The sequence and structure of the large (20s) mitochondrial (mt) rRNA gene and flanking regions from Paramecium primaurelia have been determined. The gene contains two regions of strong homology with other large mt rRNAs: one 44-base region near the 5' end and a 321-base region near the 3' end. Another region of strong homology to both ends of E. coli 23s RNA exists at loci consistent with these regions. The Paramecium gene appears to be 2204 bases in length and contains slightly more homology to E. coli rRNA than its mammalian or fungal counterparts. The gene, located about 1200 bp from the replicative terminal end of the linear mt DNA, is transcribed in the same polarity as replication. Previous R-looping studies detected no large introns within the gene. Here we describe sequences resembling degenerate rRNAs, one of which could represent a small intron. A tRNA tyr gene was found on the same DNA strand, 127 bp downstream from the large rRNA presumptive 3' end. The tRNA is flanked on both sides by short DNA regions of approximately 90% A + T content. PMID- 6275361 TI - Nucleotide sequences of the 5S ribosomal RNA genes and their adjacent regions in Schizosaccharomyces pombe. AB - The organization of 5S ribosomal RNA (rRNA) genes in the genome of Schizosaccharomyces pombe has been investigated by restriction and hybridization analyses. The 5S rRNA genes were not linked to the other three species of rRNA genes which formed a repeating unit of 6.9 megadaltons, but located in other regions surrounded by heterogeneous sequences. The 5S rRNA gene organization in S. pombe is therefore different from those in other yeasts; Saccharomyces cerevisiae and Torulopsis utilis. Four restriction segments of different sizes each containing a single 5S rRNA gene were cloned on a bacterial plasmid, and the sequences in and around the RNA coding regions were determined. In the RNA coding regions, the sequences in four clones were identical with an exception that one residue has been substituted in one clone. In the flanking regions, the sequences were extremely rich in the AT-content and highly heterogeneous. The sequences were also markedly different from those in the corresponding regions of the other two yeasts. THe presence of T-clusters in the regions immediately after the RNA coding sequences was only notable homology among the four clones and the other two yeasts. PMID- 6275360 TI - The sequence of the Drosophila melanogaster gene for yolk protein 1. AB - We have determined the complete nucleotide sequence of the hormonally regulated yolk protein 1 (YP1) gene of Drosophila melanogaster. We have also determined the sequence location of the 5' and 3' ends of both the mature mRNA and the gene's only intron. The YP1 gene contains sequences similar to those found in many other eukaryotic genes. Among these sequences are the Hogness-Goldberg box, the capping site, the ribosome binding site and the polyadenylation signal sequence, all perhaps involved in transcriptional or translational control. Also among these sequences are the consensus splice sequences. They occur at each end of the 76 nucleotide intron. One distinctive secondary structure likely to occur in either the DNA or RNA and which might be involved in the regulation of transcription or translation was also found in the YP1 gene sequence. We show the protein sequence predicted by the DNA sequence and the RNA mapping results. PMID- 6275362 TI - Analysis of transcription of the human Alu family ubiquitous repeating element by eukaryotic RNA polymerase III. AB - A series of clones that contain human Alu family elements are actively transcribed in soluble in vitro RNA polymerase III systems. The 5' ends of the in vitro transcripts are located about 170 nucleotides upstream of the eponymous Alu I site of the repeat, while a region associated with specifying of the initiation site for in vitro transcription lies in the region between 79 and 106 nucleotides upstream of the central Alu site. Thus, the RNA polymerase III transcription unit defined by the human Alu family is similar to other RNA polymerase III transcription units in possessing an internal region that is required for active transcription in vitro. PMID- 6275363 TI - 5S ribosomal RNA genes of the newt Notophthalmus viridescens. AB - The genes which code for the 5S ribosomal RNA in the newt, Notophthalmus viridescens have been cloned and analyzed. Two types of repeating unit were detected: a major type consisting of a 120 bp coding region with a 111 bp spacer, and a minor type composed of a coding region, a pseudogene, and a 113 bp spacer. The pseudogene is a 36 bp segment which corresponds to the 3' terminal third of the 5S RNA gene, and is situated immediately 3' to the gene, being separated from it by 2 bp. Two recombinant plasmids were obtained in which the major and minor units were arranged in an interspersed pattern. PMID- 6275364 TI - Release of a globin gene enriched chromatin fraction from chicken erythrocyte nuclei following DNase II digestion. AB - Mild digestion of chicken erythrocyte nuclei with deoxyribonuclease II results in the release of a chromatin fraction which is 4- to 13-fold enriched for the globin coding sequences when compared to total chicken DNA. The remaining nuclear pellet is depleted in these sequences. A maximum of 25% of the globin genes have been recovered in the released fraction. The addition of 5 mM sodium butyrate to the digestion buffer is required to obtain reproducible globin gene enrichment. The released fraction contains equimolar amounts of the four core histones and a subset of the nonhistone chromosomal proteins. The globin genes are released as large chromatin fragments which exceed the 1.6 kilobase size of the transcribed portion of the gene. PMID- 6275365 TI - Characterization and mapping of RNase III cleavage sites in VSV genome RNA. AB - Ribonuclease III cleaves the genome RNA of vesicular stomatitis virus (VSV) to yield an array of fragments which range in size from 3.5 to 0.1 x 10(6) daltons under partial digestion conditions. The locations of the RNase III cleavage sites which give rise to these fragments have been ordered relative to the 3' end of the virion RNA by digestion of 3' end-labeled RNA. Based on a map of the cleavage sites we predicted that fragments having the same size could be generated which contain information from each gene. Annealing of individual VSV mRNA probes to Northern blots of the separated RNase III-generated fragments confirmed that fragments having the same size are, in fact, generated which contain information from each coding region of the VSV genome. Analysis of maps of partial digestion products indicates that fragments having the same size arise repeatedly along the 3' half of the genome. The cleavage of VSV RNA by RNase III can be detected only if the nuclease treated molecules are denatured. This suggest that the structure features in VSV RNA which signal cleavage involve areas of higher order RNA structure. PMID- 6275366 TI - Equilibria and kinetics of lac repressor-operator interactions by polyacrylamide gel electrophoresis. AB - We describe the use of gel electrophoresis in studies of equilibrium binding, site distribution, and kinetics of protein-DNA interactions. The method, which we call protein distribution analysis, is simple, sensitive and yields thermodynamically rigorous results. It is particularly well suited to studies of simultaneous binding of several proteins to a single nucleic acid. In studies of the lac repressor-operator interaction, we found that binding to the so-called third operator site (03) is 15-18 fold weaker than operator binding, and that the binding reactions with the first and third operators are uncoupled, implying that there is no communication between the sites. Pseudo-first order dissociation kinetics of the repressor-203 bp operator complex were found to be temperature sensitive, with delta E of 80 kcal mol-1 above 29 degrees C and 26 kcal mol-1 below. The half life of the complex (5 min at 21 degrees C) is shorter than that reported for very high molecular weight operator-containing DNAs, but longer than values reported for much shorter fragments. The binding of lac repressor core to DNA could not be detected by this technique: the maximum binding constant consistent with this finding is 10(5) M-1. PMID- 6275367 TI - Nucleotide sequence of the transforming early region E1b of adenovirus type 12 DNA: structure and gene organization, and comparison with those of adenovirus type 5 DNA. AB - The nucleotide sequence of the entire transforming early region of E1b of the highly oncogenic adenovirus type 12 (Ad12) DNA has been determined. The total sequence (3860 base pairs) encompasses the entire transforming early region E1 of Ad12 DNA. From the sequence for the E1b region of Ad12, and the transcription map of the E1b region (1, 2, 3, and this paper) the structure and gene organization of the early region E1b of Ad12 DNA were analyzed and compared with those of the E1b region in the non-oncogenic Ad5 DNA (4, 5). Most of the sequences in the E1b region of Ad12 was highly homologous to that of Ad5. It is predicted that the Ad12 region E1b codes for polypeptides of 53.9, 19.1, and 8.9 kd. This situation is identical with that of the Ad5 region E1b which codes for polypeptides of 54.9, 20.6, and 8.3 kd. The function of these predicted polypeptides encoded by the E1b regions in cell transformation is discussed. PMID- 6275369 TI - Hospital diets: a question of fibre. PMID- 6275368 TI - The organization of the dihydrofolate reductase gene of baby hamster kidney fibroblasts. AB - Using cloned DNA complementary to mouse dihydrofolate reductase (DHFR) mRNA, the organization of the hamster DHFR gene has been determined in two baby hamster kidney (BHK) cell lines, A5 and B1. A5 cells are highly methotrexate-resistant, containing 200-fold more copies of the DHFR gene than do the parental B1 cells. The DHFR gene has the same organization in A5 and B1 cells, suggesting that it has not been altered by the amplification process. The BHK DHFR gene spans a maximum of 10.7 kb and contains at least three introns. Thus the BHK DHFR gene is much smaller than the mouse DHFR gene, which has a minimum size of 42 kb and at least five introns. This striking size difference is probably due to much smaller introns in the BHK DHFR gene. PMID- 6275370 TI - [Treatment of urinary tract infections with penicillin Hx]. PMID- 6275371 TI - [Usefulness of the determination of technetium -99m pyrophosphate accumulation indicators in the evaluation of the activity of rheumatoid arthritis]. PMID- 6275372 TI - [Triglyceride storage diseases]. PMID- 6275373 TI - [Bronchial asthma - is the hypothesis of adrenergic beta receptor blockade obsolete?]. PMID- 6275374 TI - Detection of normal and aberrant chromosomes in chicken embryos and in tumor cells. PMID- 6275375 TI - Chronic lead ingestion and nephropathy in ringed turtle doves. PMID- 6275376 TI - Relative efficiency of test procedures to detect lymphoid leukosis virus infection. AB - A total of 2544 comparisons were used to study the relative efficiency of various test procedures currently used to detect lymphoid leukosis virus infection in chickens. Meconium, blood, cloacal swabs, and egg albumen were collected from day old chicks and adult hens from four breeder flocks. The phenotypic mixing was used as the biological assay for infectious virus. The complement fixation test and enzyme-linked immunosorbent assays were used as direct assays for the group specific antigen of the avian leukosis-sarcoma viruses. Analysis of test results of biological assays has shown that blood from virus-infected chicks was more frequently positive than meconium. Also, direct assay for group-specific antigen was as efficient as the biological assays for detection of positive albumen samples from freshly laid eggs. The rate of false-positive reactors recognized by direct assays was lower in albumen samples than in cloacal swabs collected from hens that produce endogenous virus or group-specific antigen. Data also showed a relatively high concordance, ranging from 71 to 100%, between various assays with test materials from adult hens. Progress in reducing rates of exogenous lymphoid leukosis virus infection can be significant if dams are selected on the basis of any one of the test procedures examined. For complete eradication however, day old chicks should be selected on the basis of the biological assay of blood of individual chicks reared in small groups. PMID- 6275377 TI - [Clavine ergot alkaloids, metabolites of Penicillium gorlenkoanum]. PMID- 6275378 TI - [Use of cyproheptadine (Peritol) as a drug blocking corticotropin secretion in the treatment of Cushing's syndrome]. AB - The results are presented of the combined drug treatment of 39 patients with Icenko-Cushing's disease, using steroidogenesis inhibitor chloditane (o.p-DDD) and corticotropin secretion blocker peritol (cyproheptadine). The control group of 40 patients, suffered from the above disease of the same severity, was treated with alone chloditane. In the control group the inhibition of the adrenocortical glucocorticoid activity by chloditane was accompanied by twofold increase in the blood plasma corticotropin level. The relapse was noted in 24 of 40 patients after 5 to 6 months. The inhibition of glucocorticoid activity did not induce corticotropin secretion activation in patients, treated with chloditane and peritol. At the delayed terms of the treatment course the basal level of plasmatic corticotropin continued to lower and averaged the subnormal values. Relapses of the disease were seen in only 8 of 39 patients. The combined chloditane-perikol treatment of Icenko-Cushing's disease is likely to be perspective. PMID- 6275379 TI - Characterization of pro-opiocortin-converting activity in purified secretory granules from rat pituitary neurointermediate lobe. AB - Lysates of secretory granules from rat pituitary neurointermediate lobes were incubated with [3H]arginine- or [3H]phenylalanine-labeled toad pro-opiocortin. The processed products formed were identified by immunoprecipitation with adrenocorticotropin (ACTH) and endorphin antisera and by migration behavior on acid/urea/polyacrylamide gels. Pro-opiocortin was cleaved by the proteolytic activity in the secretory granule fraction to approximately 21,000 Mr ACTH, approximately 13,000 Mr ACTH, alpha-melanotropin, 16,000 Mr NH2-terminal glycopeptide, beta-lipotropin, and an endorphin-related peptide. Characterization of this pro-opiocortin-converting activity shows that it (i) is present in membrane and soluble fractions of the granule lysates, (ii) has a pH optimum of 5.0, (iii) appears to cleave at pairs of basic amino acid residues in the precursor, and (iv) is inhibited by leupeptin, pepstatin A, and p chloromercuribenzoate but not diisopropyl fluorophosphate, N alpha-p-tosyl-L lysine chloromethyl ketone hydrochloride, chloroquine, or EDTA. These inhibitor studies suggest that the converting-enzyme activity is due to an acid thiol, arginyl protease, distinct from any known cathepsin B-like activity. PMID- 6275380 TI - Novel sensory adaptation mechanism in bacterial chemotaxis to oxygen and phosphotransferase substrates. AB - The involvement of methylation in the chemosensory response of bacteria to many attractants has been clearly established by studies in several laboratories. It has been assumed that adaptation of Salmonella typhimurium and Escherichia coli to all attractants involves methylation of a transmembrane methyl-accepting chemotaxis protein. The methyl donor in this reaction is S-adenosyl-L-methionine, and the protein methyltransferase is the product of the cheR gene. In contrast, adaptation to oxygen and phosphotransferase substrates were found to be independent of this methylation system. In E. coli AW660 (tsr tar trg), which lacks the known methyl-accepting chemotaxis proteins, chemotaxis was normal to oxygen and to substrates of the phosphotransferase system such as D-mannose, D glucose, and N-acetyl-D-glucosamine. When S-adenosyl-L-methionine was depleted by methionine starvation or by addition of 1-aminocyclopentane-1-carboxylic acid, methylation-dependent adaptation to serine, aspartate, and ribose was defective in wild-type E. coli and S. typhimurium. However, adaptation to oxygen and phosphotransferase substrates was independent of S-adenosyl-L-methionine and the cheR product. These results suggest that there are methylation-independent and methylation-dependent mechanisms for sensory adaptation in bacteria. PMID- 6275381 TI - Nucleosome phasing and micrococcal nuclease cleavage of African green monkey component alpha DNA. AB - The micrococcal nuclease cleavage of intact nuclear chromatin from African green monkey cells and of the completely deproteinized sequences was studied by using high-resolution analytical and DNA sequencing gels and secondary restriction enzyme analysis. When deproteinized component alpha DNA was used as substrate, not all phosphodiester bonds in the 172-base-pair repeat units were cleaved with equal frequency by the nuclease. A distinct preference for the cleavage of A-T rather than G-C bonds was observed; however, A + T-richness in itself did not confer susceptibility to cleavage by micrococcal nuclease. The results suggested that, in deproteinized DNA, nuclease cleavage at particular dinucleotides may be influenced more by the effect of adjacent sequences than by the composition of the dinucleotide. In contrast to complex cleavage patterns of the deproteinized component alpha DNA which arose because of multiple cleavage sites in the repeat unit, micrococcal nuclease cleaved component alpha nuclear chromatin at one site per nucleosome repeat, near position 126 in the nucleotide sequence. This simple chromatin cleavage pattern is consistent with the discrete nucleosomal structure of component alpha in chromatin and a direct phase relationship between the component alpha DNA sequence repeats and the nucleosome protein structural repeats. PMID- 6275382 TI - Mitochondria are associated with microtubules and not with intermediate filaments in cultured fibroblasts. AB - Triple-immunofluorescence experiments with antibodies to cytochrome c oxidase, tubulin, and vimentin have been used to immunolabel the mitochondria, microtubules, and intermediate filaments inside the same cultured fibroblasts. In particular, fibroblasts were immunolabeled after they had either been transformed by infection with Rous sarcoma virus or given long-term treatment with cycloheximide. These treatments induced redistribution of the intermediate filaments into a perinuclear arrangement, segregated away from the microtubules, which remained extended to the cell periphery. In such cells, many labeled mitochondria were observed to be codistributed with the peripherally located microtubules. From these results, we infer that an association, probably involving some type of chemical linkage(s), between mitochondria and microtubules exists in these cells that is independent of the intermediate filaments. PMID- 6275383 TI - Nonrandom association of polymorphic restriction sites in the beta-globin gene cluster. AB - By using probes for epsilon-, Psibeta(1)-, and beta-globin genes, we found four additional polymorphic restriction sites that have frequencies >0.1 in persons of Mediterranean area origin, Asian Indians, and American Blacks. Three of these (HincII sites) and the two previously described polymorphic HindIII sites [one in intervening sequence (IVS) II of each gamma-globin gene] are distributed over 32 kilobases (kb) of DNA located 5' to the delta-globin gene. This region of DNA comprises two-thirds of the beta-globin gene cluster. Since each of these five polymorphic sites can be present (+) or absent (-), in theory there exist 32 possible combinations of sites (haplotypes). However, in Italians, Greeks, Indians, and Turks, 3 of the 32 haplotypes, (+----), (-+-++), and (-++-+), account for 92% of 89 beta(A) chromosomes examined. The observed frequencies for these haplotypes are 0.64, 0.15, and 0.13 in the populations studied, in contrast to expected frequencies (based on the observed gene frequencies at each of the five sites) of 0.20, 0.006, and 0.005, respectively. In American Blacks, a fourth haplotype, (----+), which is rare in non-Black populations, has a frequency of 0.37 in contrast to its expected frequency of 0.05. These results suggest a nonrandom association of DNA sequences over 32 kb 5' to the delta-globin gene in all populations studied. Two other polymorphic sites 3' to the delta gene (the newly discovered Ava II site in IVS II of the beta-globin gene and the BamHI site 3' to it) are nonrandomly associated with each other but randomly distributed with respect to the above haplotypes. This suggests that randomization of sequences has occurred within 12 kb of DNA between these two nonrandomly associated sequence clusters. Nonrandom association of polymorphic restriction sites has practical consequences in that it limits the usefulness of these additional HincII sites for prenatal diagnosis of hemoglobinopathies by linkage analysis. These sites provide little additional information for detection of beta thalassemia, while the polymorphic Ava II site, which lies outside the nonrandomly associated sequences 5' to the delta gene, improves the test applicability from 52% to 70% of couples at risk. PMID- 6275384 TI - Detection of genes coding for human differentiation markers by their transient expression after DNA transfer. AB - We have developed an assay for specific genes in DNA based on transient expression. DNA prepared from patients with acute myeloblastic or acute lymphoblastic leukemia or from the continuous leukemic cell line HL60 was transferred to LTA cells; 48-72 hr later, these recipients expressed hemopoietic differentiation markers as detected by monoclonal antibodies against My-1 (granulocyte specific) and OK-T3 (T-cell specific) using immunofluorescence. The efficiency of transfer was dose and time dependent. We found that genes not expressed in the original cells were expressed after transfer by using this assay. Restriction enzyme analysis showed that My-1 was not expressed with DNA that had been incubated before transfer with either HindIII or Sal I but was present after digestion with either EcoRI or BamHI; after digestion of DNA with the same enzymes, OK-T3 expression was observed only in the HindIII-treated DNA. These studies indicate that DNA was transferred; this approach may provide an efficient method for detecting the activity of specific genes in the absence of selection. PMID- 6275385 TI - Varicella zoster virus DNA exists as two isomers. AB - Fragments of varicella zoster virus DNA produced by EcoRI endonuclease cleavage were cloned in vector pACYC 184 and those produced by HindIII cleavage were cloned in pBR322. Restriction enzyme cleavage maps established by double digestion and blot hybridization showed that varicella zoster virus DNA has a Mr of 80 +/- 3 x 10(6) and exists as a population of two isomers. PMID- 6275386 TI - Purification of chicken intestinal receptor for 1 alpha, 25-dihydroxyvitamin D3 to apparent homogeneity. AB - The chicken intestinal 1 alpha, 25-dihydroxyvitamin D3 receptor-like protein has been purified to apparent homogeneity as determined by sodium dodecyl sulfate gel electrophoresis. The techniques employed for the purification include selective precipitation of the receptor by Polymin P (polyethyleneimine) and (NH4)2SO4 and batch adsorption to and selective elution from hydroxylapatite, followed by gel exclusion and DEAE-cellulose chromatography. Finally, the labeled receptor was eluted at a pH of approximately 6.0 on a chromatofocusing column. The protein was purified 6100-fold and the receptor was obtained in 9% yield. PMID- 6275387 TI - Benzodiazepine receptor increases after repeated seizures: evidence for localization to dentate granule cells. AB - Repeated seizures, whether induced by kindling or electroshock, result in increased numbers of benzodiazepine receptors in hippocampal formation membranes. We sought to determine the cellular constituents containing the receptor increases. Binding studies of microdissected samples localized the receptor increases to fascia dentata. [3H]Flunitrazepam autoradiographic studies showed increases of silver grain density over the granule cell and molecular layers of fascia dentata but not in other regions of hippocampal formation. Destruction of granule cells by colchicine or neonatal x-irradiation was associated with marked decrease of benzodiazepine receptor binding. Together, these results provide evidence for localization of the receptor increases to the somata and dendritic tree of the granule cells. We suggest that this cellular localization may provide a clue to the network of altered neural circuitry underlying amygdala kindling. PMID- 6275388 TI - Rat preprocarboxypeptidase A: cDNA sequence and preliminary characterization of the gene. AB - Rat carboxypeptidase A cDNA clones have been isolated from a cDNA library prepared from pancreatic mRNA. An almost complete mRNA sequence has been deduced that predicts a polypeptide having 78% amino acid sequence homology with bovine carboxypeptidase A. The amino acid sequence of the activation and signal peptides of the carboxypeptidase A precursor were inferred from the nucleotide sequence. The cDNA was used as a probe to identify DNA fragments containing carboxypeptidase A sequences in a bacteriophage lambda library of rat genomic DNA. Heteroduplexes revealed that the DNA coding sequence occupies 5.5 kilobases and is interrupted by nine intervening sequences. The nucleotide sequence of the 5' end of the gene and the adjacent flanking region provides information on the site of initiation of transcription and the putative control regions. There is no evident relationship between the localization of intervening sequences in the gene and functional/structural domains of the protein. PMID- 6275389 TI - DNA sequences affecting specific initiation of transcription in vitro from the EIII promoter of adenovirus 2. AB - We have identified those sequences affecting the level of specific initiation of transcription in vitro from the EIII promoter of adenovirus 2. Mutants containing deletions in and around the initiation sites were constructed in cloned viral DNA fragments and assayed for their ability to initiate transcription in vitro. Three classes of mutants were studied with deletions in the following regions: -38 to 268, -21 to -71 (which includes the T-A-T-A-A box), and -29 through the cap sites (+1 and +3). Deletions that remove some or all of the area from -28 to several nucleotides downstream from the cap sites essentially abolished specific transcription. Small deletions in the region -30 to -41 reduced transcription to approximately 60% of wild type; larger deletions in the region -35 to -268 reduced transcription to 30-40% of wild type. Deletions beginning from approximately +10 to +25 and extending further downstream reduced transcription to 20-40% of wild type, whereas a deletion beginning at +31 had little or no effect. Our results suggest that the region including the T-A-T-A-A box and extending to the area immediately beyond the cap sites is essential for specific transcription in vitro from the EIII promoter. However, sequences upstream from the T-A-T-A-A box and those downstream from the cap sites appear to significantly modulate the levels of transcription. PMID- 6275390 TI - Transposon-specified site-specific recombination. AB - Cointegrate DNA molecules containing two copies of a transposable element appear to be intermediates in the transposition process. These structures are resolved by site-specific recombination to yield the normal end products of transposition. The transposable element gamma delta (Tn1000) synthesizes a product interchangeable with the Tn1/3tnpR protein in promoting Tn1/3 site-specific recombination. These data support the hypothesis that cointegrates containing directly repeated copies of Tn1/3 are obligatory intermediates in interreplicon transposition of Tn1/3. In addition, we show here that the reaction is independent of the element-encoded tnpA gene product. Tn501, which specifies mercury resistance, also produces cointegrates as intermediates in interreplicon transposition. The appearance of Tn501-specified recombination activity that can act on these cointegrates requires growth of cells in the presence of Hg2+. PMID- 6275392 TI - Sizes and map positions of several plasmid-DNA-encoded transcripts in octopine type crown gall tumors. AB - The sizes and relative map positions of RNA molecules encoded by the plasmid DNA of several octopine-type tobacco crown gall tumors were determined by blot hybridization. At least six discrete polyadenylylated RNAs (sizes 0.73-1.75 kilobases) were detected in octopine-producing tumors (lines A6S/2, E9, and 15955/01). In one tumor (line 15955/1) in which octopine could not be detected, one of the RNA species was missing and another was diminished in abundance. Evidence is presented suggesting that the transcripts are encoded entirely by the plasmid DNA and do not originate from the surrounding plant DNA sequences. PMID- 6275391 TI - Nucleotide sequence and the encoded amino acids of human serum albumin mRNA. AB - The complete nucleotide sequence of human serum albumin mRNA has been determined from recombinant cDNA clones and from a primer-extended cDNA synthesis on the mRNA template. The sequence is composed of 2078 nucleotides, starting upstream from a potential ribosome binding site in the 5' untranslated region. It contains all the translated codons and extends into the poly(A) at the 3' terminus. Part of the translated sequence codes for a hydrophobic prepeptide, Met-Lys-Trp-Val Thr-Phe-Ile-Ser-Leu-Leu-Phe-Leu-Phe-Ser-Ser-Ala-Tyr-Ser, followed by a basic propeptide, Arg-Gly-Val-Phe-Arg-Arg. These signal peptides are absent from mature normal serum albumin and, so far, have not been identified in their nascent state in humans. A remaining 1755 nucleotides of the translated mRNA sequence code for 585 amino acids, which are in agreement, with few exceptions, with the published amino acid sequence for human serum albumin. The mRNA sequence verifies and refines the repeating homology in the triple-domain structure of the serum albumin molecule. PMID- 6275394 TI - Running elevates plasma beta-endorphin immunoreactivity and ACTH in untrained human subjects. AB - Twenty minutes of submaximal treadmill running was associated with an elevation in plasma levels of beta-endorphin immunoreactivity (P less than 0.02). This increase was greater in men (14.9 +/- 3.4 fmole/ml) than women (2.6 +/- 1.2 fmole/ml)(P less than 0.05). Plasma levels of ACTH and growth hormone also increased after running. ACTH increased more in men (7.8 +/- 1.1 fmole/ml) than in women (1.1 +/-0.44 fmole/ml)(P less than 0.02). There was a similar growth hormone response in both sexes. No correlation can at this time be made with levels in the central nervous system. Changes in plasma levels of beta-endorphin immunoreactivity may be responsible for some of the euphoria and analgesia anecdotally associated with running. PMID- 6275393 TI - Structure of two human alpha-tubulin genes. AB - The ability of a chicken alpha-tubulin cDNA probe to cross-hybridize with human DNA under stringent conditions has been exploited to screen two independently constructed human genomic libraries. Nine clones were isolated, accounting for 60% of the bands observed in a whole genomic Southern blot of human DNA. Two clones were selected for further analysis by restriction mapping, orientation experiments using 3'- or 5'-specific probes, and electron microscopy of heteroduplexes. One clone, 2 alpha, contains an alpha-tubulin-specific region of 5.0 kilobases that includes three intervening sequences. The second clone, 19 alpha, contains an alpha-tubulin-specific region of 5.4 kilobases and has somewhat diverged 5' and 3' ends. Clone 19 alpha has only two intervening sequences that correspond to the first two in clone 2 alpha. However, these intervening sequences differ in size between clones 2 alpha and 19 alpha and show no detectable sequence homology. The sum of the lengths of sequences in either clone that hybridize to the cDNA probe accounts for essentially the entire length of the cDNA molecule. PMID- 6275395 TI - Diurnal influences on serum luteinizing hormone responses to opiate receptor blockade with naloxone or to luteinizing hormone-releasing hormone in the immature female rat. PMID- 6275396 TI - Down-regulation of prolactin receptors in rabbit mammary gland: differential subcellular localization. PMID- 6275397 TI - Mechanistic studies of the gastric (H+ + K)-ATPase. PMID- 6275398 TI - Electrogenic Na-K antiport and electrogenic Na-C1 symport in basolateral membrane of frog stomach. PMID- 6275399 TI - Hormonal control of the Na entry process at the apical membrane of frog skin. PMID- 6275400 TI - The rheogenic nature of sodium transport in leaky epithelia studied by equivalent circuit analysis. PMID- 6275401 TI - Current studies on the mechanisms underlying the onset of female puberty. PMID- 6275402 TI - Role of prostaglandins in the control of pituitary hormone secretion. PMID- 6275403 TI - Ectopic production of growth hormone-releasing factor by carcinoid and pancreatic islet tumors associated with acromegaly. PMID- 6275405 TI - Regulation of catecholamine secretion by endogenous opioid peptides. PMID- 6275404 TI - Antifertility effects of luteinizing hormone-releasing hormone (LHRH) agonists. PMID- 6275406 TI - Polymorphism of estrogen receptors in human breast cancer. PMID- 6275408 TI - Regulation of thyrotropin secretion by 3,5,3'-triiodothyronine and thyroxine. PMID- 6275407 TI - Receptors, enzymes, and hormonal responses of endometrial cells. AB - Studies of hormone-related biochemical parameters of human endometrium are facilitated by the availability of a variety of types of endometrial tissue resulting from changes in hormonal environment during the menstrual cycle or from decidual, hyperplastic, and neoplastic transformations. Estrogen and progesterone receptors levels, rates of metabolism of ovarian hormones, enzymatic activities regulated by hormones or affecting intracellular concentrations of steroids, rates of production of prolactin and prostaglandins, and endogenous levels of hormones have been measured in samples of endometrial tissue and in separated glands and stroma. These studies have also included an evaluation of effects of estrogens and progestins administered to patients or added to the medium during in vito incubations of endometrium under organ culture conditions. Epithelial cells derived from isolated glands and stromal cells have been grown in primary cultures and also subcultured. These studies have revealed two mechanisms by which progestins exert an antiestrogenic effect on the endometrial tissue: viz, they were found to provoke a decline in estrogen receptor levels and to increase the metabolism of E2 through stimulation of estradiol 17 beta-dehydrogenase activity. In addition, studies of cells in culture have led to the unexpected finding of wide and rapid fluctuations in the levels of estrogen receptors, thus providing an opportunity to investigate factors involved in the regulation of concentrations of specific E2-binding sites. PMID- 6275409 TI - Relation of prolactin to development of spontaneous mammary and pituitary tumors. PMID- 6275410 TI - Chlorinated insecticides and hormone receptors of the prostate. PMID- 6275411 TI - Inhibitors of prolactin secretion: a mini-review. PMID- 6275412 TI - Retinoic acid receptor and surface markers: models for the study of prostatic cancer cells. PMID- 6275413 TI - The development of gamma-emitting hormone analogs as imaging agents for receptor positive tumors. PMID- 6275414 TI - Polypeptide hormones and the prostate. AB - Results presented in this paper demonstrate that both insulin and prolactin have direct effects on the growth and functions of prostatic epithelium. They also act on the prostate indirectly by potentiating the effects of testosterone on these cells. Both are capable of inducing cell multiplication and increasing the activity of ornithine decarboxylase in cells. However, the intracellular events which follow the interaction of these hormones with the cell membrane receptors are not clearly understood. Little is known at the present time about the specific effects of insulin and prolactin on prostatic growth regulation and function and about their interactions with other protein and steroid hormones. A considerable amount of further research is needed to establish the basic influences of insulin and prolactin on the prostate. This inquiry is essential before the role of these hormones in normal and abnormal growth of the prostate can be understood. PMID- 6275415 TI - Visualization of prolactin binding sites in prostate tissue. PMID- 6275416 TI - The role of prolactin in the growth of the prostate gland. PMID- 6275417 TI - An open, multi-centre evaluation of a 5-day course of pivmecillinam in urinary tract infections in general practice. AB - One hundred and eleven general practice patients (101 females and 10 males) with acute uncomplicated cystitis received a 5-day course of pivmecillinam at a daily dose of 200 mg 3-times a day. Positive bacteriological cultures were found in 63 (57%) patients before treatment and three-quarters of all patients were totally symptom-free after treatment, with the post-treatment clinical symptom score falling by 94%. In those patients with a positive culture the bacteriological cure rate was 90%. Pivmecillinam was generally well tolerated. Side-effects were reported in 8 (7%) patients and only 1 of these patients had to stop treatment prematurely. PMID- 6275418 TI - Method of sacrifice influences leucine enkephalin binding to mouse brain. AB - The methods used for sacrifice of animals and the time taken for any dissection procedures of the brain tissue at room temperature have been shown to influence the number of available binding sites for [3H] leucine enkephalin (10 nM) to mouse brain homogenates. These factors should be considered when utilizing receptor binding techniques as an indirect method for evaluating the in vitro functional state of opiate receptor populations. PMID- 6275419 TI - The correlation between swim-stress induced antinociception and [3H] leu enkephalin binding to brain homogenates in mice. AB - Mice which had been made to swim for 3 minutes showed a tail flick latency which was significantly longer than that of unswum controls. The [3H] leu-enkephalin [LE] binding to brain homogenates from swum mice was significantly reduced when compared with that form unswum controls. Scatchard analysis revealed that the reduction in binding occurred at the LE low affinity site. However, when homogenates were allowed a preincubation period of 20 min at 37 degree C, the difference in LE binding between swum and unswum mice was no longer apparent. These data are interpreted to suggest that the reduced LE binding may be due to the occupation of a proportion of the opiate receptor population by an endogenous ligand. A correlation between the duration of the swim induced antinociceptive response and the changes in LE binding is described which although non significant, is consistent with the interpretation for the involvement of endogenous opiates in the observed increases in tail flick latency. PMID- 6275420 TI - The effects of acute administration of diazepam on the binding of [3H]-diazepam and [3H]-gaba to rat cortical membranes. AB - Specific [3H]-diazepam binding and [3H]-GABA binding were measured in cortical membranes of untreated rats and rats which had been administered unlabeled diazepam (5.0 mg/kg, IP) thirty minutes prior to sacrifice. Washed and unwashed membranes from control animals showed identical levels of [3H]-diazepam binding. Unwashed membranes of diazepam-treated animals showed consistently and significantly lower binding of [3H]-diazepam than membranes derived from control animals and treated similarly. [3H]-GABA was almost non-existent in unwashed membranes of either group of animals. The binding capability of membranes of treated animals for [3H]-diazepam returned to control levels upon washing with buffer prior to the binding assay. The specific binding of [3H]-GABA in membranes derived from either group of animals also improved after the buffer washes. However, no difference could be detected in [3H]-GABA binding between control and diazepam-treated animals. The failure of diazepam to modulate [3H]-GABA binding in unwashed membranes and the participation of an endogenous inhibitory material repressing [3H]-GABA binding are discussed. PMID- 6275421 TI - Adrenalectomy reverses the effects of delta-9-THC on mouse brain 5 hydroxytryptamine turnover. AB - Acute administration of certain cannabinoids, including delta 9 tetrahydrocannabinol (delta 9- THC), resulted in elevated levels of plasma corticosterone in mice. The rank order potency of these cannabinoids is the same as others have reported using behavioral tests. The maximally effective dose of delta 9-THC (30 mg/kg) in this test also increased the amount of 3H-tryptophan found in the brains of mice given an intravenous injection of 3H-tryptophan 10 min prior to decapitation. This effect was associated with an increase in the amount of 3H-5-hydroxytryptamine synthesized during the pulse period. Adrenalectomy was found to inhibit these effects of delta 9-THC. The possibility that corticosterone may mediate the effects of delta 9-THC on tryptophan disposition and metabolism is discussed. PMID- 6275422 TI - Anti-inflammatory mechanism of prozime-10, a proteolytic enzyme. AB - When Prozime-10 (P-10), a protease extracted from cultured both of Aspergillus melleus, was injected intravenously into anesthetized dogs, plasma ACTH was increased with a latency of 30 min, and this was followed by remarkable elevation of plasma cortisol in many instances. A similar increase in plasma cortisol was elicited after trypsin and alpha-chymotrypsin were injected. Plasma histamine was raised promptly prior to an increase in plasma ACTH after P-10 in every case. However, in certain cases, changes in cortisol occurred simultaneously with ACTH after P-10. Such a rapid elevation of cortisol can be explained, partly, by direct stimulation of the adrenal cortex by histamine. PMID- 6275423 TI - Endogenous ligands and modulators of the digitalis receptor: some candidates. PMID- 6275424 TI - Computed tomography using proton energy loss. PMID- 6275426 TI - Environmental enrichment to alleviate maze performance deficits in rats with microcephaly induced by x-irradiation. PMID- 6275425 TI - Physiological implications of the presence, distribution, and regulation of calmodulin in eukaryotic cells. PMID- 6275427 TI - Effects of malnutrition, maternal stress, or ACTH injections during pregnancy on sexual behavior of male offspring. PMID- 6275428 TI - Effect of a short-term fasting and altitude hypoxia on the cytochrome oxidase activity of rat brain mitochondria. AB - The effect of short-term fasting and thirst, prolonged fasting and hypoxic hypoxia upon the activity of cytochrome oxidase was studied in mitochondrial fractions obtained from the brain and the liver. The investigation was carried out in two groups of rats, 5 and 60 days old. a) The activity of cytochrome oxidase in mitochondria isolated from the brain cortex, subcortical regions and the medulla oblongata rises, while the changes in liver mitochondrial fractions are reverse. b) A significant increase of mitochondrial cytochrome oxidase was found in 5-day-old rats after both types of fasting and hypoxia in all regions of the brain, as well as in the liver. c) The cytochrome oxidase activity in brain and liver mitochondria of 60-day-old rats was not affected appreciably after 24 h nutritional deprivation, with the exception of a significant rise of activity in the medulla oblongata. Prolonged fasting and hypoxia again markedly increased the activity of this enzyme in all regions of the brain and in the liver. PMID- 6275429 TI - Comparison between grafts with intact nerves and standard free grafts of the rat extensor digitorum longus muscle. AB - Standard grafts and nerve-intact grafts of the extensor digitorum longus muscle were compared in the rat. In standard grafts the muscle was completely removed from its bed and replaced; nerve-intact grafts were treated in an identical manner except that the muscle nerve was not severed. Nerve-intact grafts underwent the same sequence of skeletal muscle fibre degeneration and regeneration as standard grafts. In nerve-intact grafts the intramuscular portions of the nerve fibres initially degenerated, but within a week new nerve fibres had regenerated back to the original zone of motor end-plates. By 60 days the weight of nerve-intact grafts approached those of control muscles. Contractile tension in nerve-intact grafts was greater than that of standard grafts. In standard and nerve-intact grafts choline acetyltransferase activity rapidly decreased to low values and then increased along curves roughly paralleling the muscle weights. In nerve-intact grafts, neuromuscular transmission was established early in the second week whereas a considerably later return was seen in standard grafts. Either the early onset or the topographical pattern of reinnervation are potentially major factors in determining the success of free muscle grafts. PMID- 6275430 TI - Functional properties of muscle autografts substituted for the rat levator ani muscle. AB - The present report deals with the functional properties (contraction parameters and neuromuscular transmission) of muscle grafts and transposed muscles substituted for the levator ani muscle in the rat. The experiments were divided into four main groups. Group I - the levator ani [LA] was excised and replaced in its own bed. Group II - the extensor digitorum longus, a fast muscle (with or without predenervation), and Group III - the soleus, a slow twitch muscle, were substituted for the LA. In group IV, the gracilis anterior muscle was either freely grafted in place of the LA or transposed a) with intact innervation, b) with its vascular supply intact or c) with preserved neuro-vascular supply. The optimum results of twitch and tetanic tension, and the amplitude of stimulation EMG responses was found in the case of LA resutured into its own bed and in the case transposition of the gracilis anterior muscle had been performed with its neuro-vascular supply intact in place of the LA. On the basis of these functional findings and morphological and anatomical observations (Grim et al. 1982), a surgical procedure is suggested for patients with anal incontinence (Grim et al. 1981, Dittertova-Vlasakova et al. 1982). PMID- 6275431 TI - Digoxin-like immunoreactivity of certain steroid and other hormones. PMID- 6275432 TI - Extramammary Paget's disease. AB - Extramammary Paget's disease is an in situ skin and mucosal carcinoma frequently associated with and probably arising in a subjacent or regionally proximate carcinoma. Microscopic spread of tumor cells almost always extends beyond clinically apparent disease. Surgical treatment requires carefully planned, systematic excision under precise histologic control. An ideal treatment method remains to be developed. Inadequate excision usually results in recurrences that can be successfully treated by reexcision. Associated invasive carcinomas occur frequently, and mortality is high in these patients. PMID- 6275433 TI - Teres major--latissimus dorsi skin-muscle flap for chest-wall reconstruction. AB - The complete survival of a teres major--latissimus dorsi skin-muscle flap for chest reconstruction is reported in a patient in whom the primary neurovascular pedicle of the latissimus muscle had previously been sacrificed. At operation, collateral vessels of 1-mm proportions were noted to have developed between the innervated teres major and denervated latissimus dorsi muscle. In this case, the teres major muscle rotated with the latissimus muscle to affect reconstruction of a full-thickness anterior chest resection. This report suggests the possibility of a denervated, relatively devascularized muscle obtaining additional blood supply from adjacent innervated vascularized muscles and supports the concept of rotating the muscles as a unit to encourage viability of the denervated component. PMID- 6275434 TI - Cutaneous silica granuloma. AB - Cutaneous silica granuloma is a rare and fascinating cause of delayed scarring. Symptoms may arise many years after minor wounding if glass, sand, or other silica-containing particles have been embedded. Such granulomatous lesions are often nodular, indurated, and erythematous. They may respond to surgical excision or medical treatment, and in rare cases, they may spontaneously regress. We present one case in which three separate areas of cutaneous silica granulomas developed after a latent period of about 20 years and then involuted without intervention. The etiology and therapeutic options are discussed. PMID- 6275435 TI - In vivo effects of leukotriene B4, C4 and D4. Evidence that changes in blood pressure are mediated by prostaglandins. AB - Intra-jugular nanomole injections of leukotrienes B4, C4 and D4 (LTB4, LTC4, LTD4) in anesthetized guinea-pigs have been shown to cause dose-dependent increases of the mean arterial blood pressure. While the responses to LTB4 were monophasic, the responses to LTC4 and LTD4 were characterized by a fast (10-50 sec), medium high, first pressor phase followed by a second, longer lasting (3-9 min), more important pressor phase. Like antigen-antibody reactions, leukotrienes induced cardiac effects such as tachycardia and rhythm disturbances as well as respiratory difficulties, convulsions and sometimes death of the animals. The prostaglandin synthesis inhibitor, indomethacin, reduced the pressor response and the tachyarrhythmic effects of LTB4, C4 and D4. These results raise the possibility that leukotrienes produce their hemodynamic effects in guinea-pigs by stimulating the synthesis and release of biologically active derivatives of arachidonic acid. PMID- 6275436 TI - CSF Vasopressin and cyclic nucleotide concentrations in senile dementia. PMID- 6275437 TI - Effect of an ACTH 4-9 analog on human cortical evoked potentials in a constant foreperiod reaction time paradigm. PMID- 6275438 TI - beta-Endorphin and beta-lipotropin plasma levels in chronic schizophrenia, primary affective disorders and secondary affective disorders. PMID- 6275439 TI - A repeated test procedure to assess onset and duration of the cue properties of ( ) delta 9-THC, (-) delta 8-THC-DMH and (+) delta 8-THC. AB - Rats were trained in a two-lever operant box in drug discrimination procedure to respond differentially to the effects induced by 3 mg/kg of (-) delta 9 tetrahydrocannabinol (delta 9-THC) and the drug vehicle. Tests with (-) delta 9 THC and the dimethyl-heptyl (DMH) homologue of (-) delta 8-THC indicated that (-) delta 8-THC-DMH was more potent but had a slower onset of action than (-) delta 9 THC. Two ways of testing the onset and duration of action were compared. In one procedure (separate tests) the time course of the drug action was established by testing each time interval on separate days with a new injection each test day, whereas in the other procedure (repeated tests) all intervals were evaluated after a single injection. The results were similar for both procedures. The median time intervals for the decay of the (-) delta 9-THC stimulus were 122 and 127 min for the separate and repeated tests procedures respectively. The median time intervals for the onset of action of the (-) delta 9-THC-like effects of (-) delta 8-THC-DMH were 65 and 62 min for the separate and repeated tests procedures respectively. The median time intervals for the decay of (-) delta 8-THC-DMH (0.30 and 0.56 mg/kg) was between 8 and 24h after injection. Furthermore, a stereoselective action is indicated, as (+) delta 8-THC (5.6 and 10 mg/kg) did not substitute for (-) delta 9-THC. PMID- 6275440 TI - Relationship between plasma delta-9-tetrahydrocannabinol concentration and pharmacologic effects in man. AB - The relationship between each of two pharmacologic effects (tachycardia and psychological "high") of delta-9-tetrahydrocannabinol (THC) and plasma THC concentration was investigated in three male and three female experienced marihuana smokers. Each subject smoked 1% THC cigarette on two occasions separated by 2 h. Heart rate and subjective psychological self-rating were determined frequently throughout the 4 h study period. Data were analyzed by calculating the area under the parameter versus time curves, constructing hysteresis plots, and calculating the decay rate constants from pharmacologic effect versus time plots. In both males and females, dose inhaled and psychological response were apparently equivalent for the first and second cigarettes. While all subjects exhibited marked tachycardia in response to the first cigarette, heart rate in both male and female subjects was not increased as markedly during the second cigarette. Interestingly, female subjects had less tachycardiac response than males during the second cigarette. Hysteresis plots revealed that both heart rate and subjective psychological effect were elicited in an effect compartment which was "deep" relative to the reference plasma compartment. The time courses of tachycardiac and psychological responses lagged behind the plasma THC concentration-time profile. Zero-order decay rate constants for subjective psychological rating did not change substantially from the first to second cigarette. This study suggest that plasma THC concentration is a poor predictor of simultaneously occurring physiological and psychological pharmacologic effects. PMID- 6275441 TI - Subsensitivity of human beta-adrenergic adenylate cyclase after salbutamol treatment of depression. AB - Although numerous studies have suggested that depression may be associated with a reduction in synaptic noradrenaline in the brain, direct beta-adrenergic receptor agonist have only recently been tested in the treatement of depression. Moreover, newer theories of antidepressant action suggest that a reduction in beta adrenergic receptor sensitivity is a better correlate of antidepressant treatment than noradrenaline turnover changes. Eleven depressed patients were treated with salbutamol, a beta-2-adrenergic agonist, and beta-2-adrenergic receptor sensitivity was evaluated before, during, and after treatment. beta-Adrenergic receptor sensitivity was evaluated by measuring the plasma cyclic AMP increase after an IV dose of salbutamol. The beta-adrenergic agonist exhibited antidepressant efficacy and induced subsensitivity of the beta-adrenergic adenylate cyclase with a time course paralleling the antidepressant effects. The results support the concept that receptor sensitivity changes occur during antidepressant therapy. PMID- 6275442 TI - Anticonvulsant doses of inosine result in brain levels sufficient to inhibit [3H] diazepam binding. AB - Several purines have been shown to be competitive inhibitors of [3H] diazepam binding. Inosine has also been shown to have benzodiazepine-like neurophysiologic, pharmacologic and behavioral effects, and to partially inhibit caffeine-induced seizures in mice. Using presumptive therapeutic doses of inosine, levels were determined in mouse brain at various times following injection. Inosine and hypoxanthine concentrations in brain increased several fold following inosine administration, indicating that inosine permeated the blood-brain barrier. The levels of inosine and hypoxanthine attained in brain were sufficient to inhibit by more than 50% the GABA-stimulated [3H] diazepam binding. These data suggest that the anticonvulsant properties of inosine are related to its interaction with the benzodiazepine receptor. PMID- 6275443 TI - Gnawing and changes in reactivity produced by microinjections of picrotoxin into the superior colliculus of rats. AB - Lesions of the superior colliculus in rats attenute the oral stereotypy produced by systemic administration of dopamine agonists. Current evidence suggest that such drugs affect the superior colliculus by reducing transmission in the inhibitory GABAergic nigrotectal pathway. To investigate whether the superior colliculus plays a direct role in producing stereotyped oral movements, the present experiment therefore examined the effects of collicular microinjections of the GABA antagonist picrotoxin on the behaviour of rats observed in an open field. Gnawing was observed after injections of picrotoxin (25 ng) into sites in the intermediate and deep layers of the superior colliculus, consistent with the superior colliculus playing a direct part in producing the stereotyped gnawing seen after systemic administration of dopamine agonists. However, gnawing was only observed after a period in which the animal showed strong avoidance reactions to stimuli that normally evoked orienting or little reaction. This change in reactivity was observed after injections of picrotoxin into all parts of the colliculus, but the most sensitive (responding to doses as low as 12.5 ng) were mainly in the superficial and intermediate layers. It appears that there may be more than one GABAergic system within the superior colliculus. PMID- 6275444 TI - Collicular picrotoxin alleviates akinesia but not sensory neglect in rats with bilateral 6-hydroxydopamine lesions of ventral midbrain. AB - Anatomical and biochemical investigations have suggested that GABA transmission in the superior colliculus consequent upon activity of the nigrotectal pathway is increased following 6-hydroxydopamine (6-OHDA) lesions of the ascending dopamine systems. Moreover, it has been proposed that this increase in inhibitory activity within the colliculus may be responsible for the sensory neglect commonly observed after dopamine denervation. The present experiment sought to test this proposal by examining the effects of injections of the GABA antagonist picrotoxin into the superior colliculus of 6-OHDA lesioned rats, in the hope that the neglect caused by the 6-OHDA would be reversed. However, in 33 of 36 cases studied intracollicular microinjections of picrotoxin produced no detectable improvement in orientation to sensory stimuli, although a wide range of other behavioural effects was observed. These included stereotyped exploratory movements (e.g. head waving, walking, sniffing and rearing) similar to those produced in the 6-OHDA treated rats by systemic injection of apomorphine (0.1 mg/kg). These data indicate that 6-OHDA lesions of substantia nigra and ventral midbrain areas do not produce sensory neglect simply by increasing GABA transmission within the nigrotectal pathway. Instead, such an increase in nigrotectal activity may impair production of particular kinds of movement, possibly related to exploratory behaviour. PMID- 6275445 TI - Nonspecific excitatory effects of morphine: reverse-order precipitated withdrawal and dose-dose interactions. AB - We recently reported that, in naive mice pretreated with naloxone, morphine can cause withdrawal-like signs that seemingly are not mediated by "opiate" receptors. Such results were confirmed and extended here with another mouse strain. Repetitive vertical jumping could occur respective of injection sequence and depended on dose and dose ratio of the two drugs. PMID- 6275446 TI - Effects of ovariectomy, castration, and chronic lithium chloride treatment on stereotyped behavior in rats. AB - The effects of ovariectomy, castration, and chronic lithium chloride treatment on stereotyped behavior (SB) induced by apomorphine (APO) (0.3--0.6 mg/kg) were investigated in rats. Duration of stereotyped behavior (interval between APO injection and termination of SB) increased in ovariectomized rats compared to female control rats. Castration had no effect on the duration of stereotyped behavior. On the other hand, ovariectomized rats that were treated chronically with LiCl (2mEq/kg daily) showed no difference in duration of stereotyped behavior compared to female controls chronically treated with LiCl. Neither treatment group showed a significantly altered intensity of stereotyped behavior compared with the appropriate control. These findings are consistent with the hypothesis that estrogen deficit contributes to a greater incidence of neuroleptic-induced tardive dyskinesia in postmenopausal women than in men of comparable age. Furthermore, LiCl may attenuate the symptoms associated with increased postsynaptic dopamine receptor sensitivity following ovariectomy. PMID- 6275448 TI - Relaxation studies of ion transport systems in lipid bilayer membranes. PMID- 6275447 TI - Excitatory and depressant effects of delta 9-tetrahydrocannabinol and cannabidiol on cortical evoked responses in the conscious rat. AB - The influences of delta 9-tetrahydrocannabinol (THC) and cannabidiol on electrically evoked cortical potentials of conscious rats with chronically implanted electrodes were investigated. Specifically, the cannabinoids' effects on a transcallosal evoked response were compared with those of ethosuximide, phenytoin, and pentylenetetrazol. THC produced dose-related opposite effects: Low doses increased the amplitude of the response, whereas higher doses reduced the response. Other drugs that can cause or exacerbate seizures, i. e., phenytoin and pentylenetetrazol, also increased the amplitude of the cortical response. In contrast, cannabidiol, over a wide dosage range, caused only depression. Ethosuximide, like cannabidiol, elicited a depressant effect. The data indicate that under the conditions of the present investigation, cannabidiol shares electrophysiological properties with ethosuximide but not with phenytoin, and that cannabidiol is a relatively selective, centrally acting drug. In addition, our findings support the suggestion that augmentation of neurotransmission in central pathways may contribute to the convulsant actions of THC, and the cannabinoids' depressant effects may, at least partially, account for their anticonvulsant actions. PMID- 6275449 TI - [Concept of the stochastic structure of the track of a heavy charged particle in matter]. PMID- 6275450 TI - [Potentiation of the protective action of sulfur-containing radioprotectors]. PMID- 6275451 TI - [Changes in rat liver adenylate kinase activity in radiation sickness and the administration of ATP to irradiated animals]. PMID- 6275452 TI - [Enzymatic activity of epithelial colonies of the small intestine and erythroid endocolonies after irradiation]. PMID- 6275454 TI - Catecholamines and the sodium pump in excitable cells. PMID- 6275453 TI - Intra-arterial chemotherapy of malignant fibrous histiocytoma of the pelvis. AB - In 3 cases of malignant fibrous histiocytoma of the bony pelvis, intra-arterial chemotherapy was followed by surgical resection when possible in an attempt to control local disease without unacceptable deformity and treat undetected systemic metastases. Cis-diamminedichloroplatinum (CDDP) and a combination of intra-arterial doxorubicin, 5-(dimethyltriazeno)imidazole-4-carboxamide, and intravenous cyclophosphamide (CYADIC) were used. In 2 patients who underwent surgical resection, no viable tumor cells were found. The third patient did not respond to these or other forms of treatment. The patients treated with resection have shown no evidence of local recurrence at 24 and 9 months, respectively. PMID- 6275455 TI - Effect of precursors of the 1, 2, and 3 series prostaglandins on renin release and renal blood flow in the dog. AB - The precursors of the monoene, diene, and triene series of prostaglandins, eicosatrienoic acid, arachidonic acid, and eicosapentaenoic acid, respectively, were infused at 3 X 10-6, 10-5, and 3 x 10-5 g/kg/min directly into the renal artery of non-filtering, denervated kidneys of conscious propranolol-treated dogs. Renal blood flow was measured with an electromagnetic flow probe around the renal artery and renal renin secretion rate from blood samples taken from catheters in the aorta and renal vein. The highest dose of arachidonic acid increased renal blood flow by 54 +/- 19% and increased renin secretion rate seven fold. Eicosatrienoic acid produced a smaller increase in renal blood flow but did not significantly increase renin secretion rate. Eicosapentaenoic did not change either blood flow or renin secretion rate. We conclude that compared with arachidonic acid the precursors of the 1 and 3 series of prostaglandins are not significantly involved in the regulation of renal blood flow or renin secretion. PMID- 6275456 TI - Evidence for a dual action of converting enzyme inhibitor on blood pressure in normal man. AB - We studied the effect of a converting enzyme inhibitor (CEI), Captopril SQ 14,225 50 mg p.o. in eight supine normal subjects under a high sodium (150 mEq/d) and low sodium (25 mEq/d) diet. On high sodium, plasma renin (PRA) and aldosterone were basal and Saralasin did not lower mean blood pressure. However, CEI induced an 11.4 +/- 3.2 mm fall in blood pressure (p less than 0.02) and either indomethacin 50 mg or ibuprofen 800 mg (PI), when given simultaneously on another day abolished the blood pressure response (2.5 +/- 0.9 mm Hg, p greater than 0.5). In contrast, on a low salt diet where renin was increased, CEI induced a drop in blood pressure which was not significantly altered by PI (12.8 +/- 1.1 vs. 10.0 +/- 3.1 mm Hg, p greater than 0.5). CEI increased plasma renin on both diets (1.7 +/- 0.5 to 3.5 +/- 0.8 and 2.8 +/- 0.6 to 12.5 +/- 3.1 ng/ml/hr respectively both p less than 0.05). Aldosterone did not change (high Na+) or fell (low Na+). Inhibition of Prostaglandin synthesis did not significantly block the renin rise from CEI suggesting that the direct angiotensin II negative feedback is relatively independent of acute prostaglandin release. Our studies suggest that CEI has a dual hypotensive action. In a low renin state, the hypotensive action appears to be mediated through vascular prostaglandins. PMID- 6275457 TI - Actions of indomethacin and prostaglandins E2 and D2 on nerve transmission in the nictitating membrane of the cat. AB - The effects of PGE2, PGD2 and cyclooxygenase inhibitors on sympathetic nerve transmission in the nictitating membrane of anesthetized cats were studied to further characterize the actions of these prostaglandins and to define their possible role as neuromodulators. PGD2 depressed neurotransmission throughout a broad range of stimulus free quencies, apparently by presynaptic inhibition of norepinephrine release. PGE2 enhanced the effects of both nerve stimulation and exogenous norepinephrine in intact preparations but only depressed the effects of exogenous norepinephrine in the chronically denervated nictitating membrane, suggesting that part of the effect of PGE2 on neurotransmission was presynaptic. When norepinephrine reuptake was blocked by desipramine, PGE2 still enhanced neurotransmission. Indomethacin, but not other cyclooxygenase inhibitors (aspirin, ibuprofen, flurbiprofen, meclofenamic acid), inhibited the response of the nictitating membrane to nerve stimulation without depressing the effects of exogenous norepinephine. Curiously, indomethacin, but again not other cyclooxygenase inhibitors, specifically antagonized the ability of PGE2 to enhance nerve transmission. These results further characterize the pharmacological effects of PGE2 and PGD2 at the nictitating membrane. The lack of effect of cyclooxygenase inhibitors suggests that neither endogenous PGE2 or PGD2 play a functional role at this synapse. The effects of indomethacin appear to be unrelated to inhibition of prostaglandin synthesis. PMID- 6275459 TI - [Detection of IgM antibody to hepatitis A virus by HAVAB-M RIA kit (author's transl)]. PMID- 6275458 TI - Arachidonate metabolism in the mouse thyroid implication of the lipoxygenase pathway in thyrotropin action. AB - The present study of compares the effects of various inhibitors of arachidonate metabolism on mouse thyroid cyclo-oxygenase and lipoxygenase activities and thyrotropin-augmented cyclic-AMP accumulation. Mouse thyroid homogenate converts [1-14C]- arachidonate to several products of the cyclo-oxygenase pathway as well as one major product of the lipoxygenase pathway, 12-L-hydroxyeicosatetraenoic acid (12-Hete). Prostaglandin (PG) formation in thyroid homogenates is inhibited by 1-10 microM indomethacin and etya. 12-HETE accumulation is reduced by 91%, 83% and 20% by 5 microM ETYA, 15-HETE, and indomethacin, respectively. Thyrotropin stimulated cyclic-AMP accumulation, measured in whole thyroid lobes by radioimmunoassay, is reduced by 45% and 73% by 50 microM and 100 microM ETYA, respectively; indomethacin is without effect at these concentrations. 15-HETE reduces thyrotropin-augmented cyclic-AMP accumulation by 57% and 100 microM. In product inhibition studies, 10 microM 12-HETE reduced the formation of radiolabeled 12-HETE by 20%. 10 microM PGE2, PGF2 alpha or PGD2 had no effect on [1-14C]-PG formation. 12-HETE, however, reduced PG synthesis by 76% at 10 microM. This is the first report implicating the arachidonate lipoxygenase pathway in thyrotropin action at the level of cyclic-AMP regulation. Additionally, our finding that 12-HETE inhibits prostaglandin synthesis suggests that the cyclo oxygenase and lipoxygenase pathways in the mouse thyroid may be highly integrated. PMID- 6275460 TI - Diverse applications of radioimmunoassay. PMID- 6275461 TI - [Response of several adrenal steroids to rapid ACTH stimulation test in essential hypertension and primary aldosteronism (author's transl)]. AB - Plasma concentration of progesterone (P), 17 alpha-hydroxyprogesterone (17 alpha P), 11-desoxy-cortisol (S), cortisol (F), 11-desoxycorticosterone (DOC), corticosterone (B), aldosterone (ALD), dehydroepiandrosterone-free (DHA-f), dehydroepiandrosterone-sulfate (DHA-s) and testosterone (TEST) were measured in 26 essential hypertension (E.H.) and 6 primary aldosteronism (P.A.) before, 30 min. and 60 min. after the intravenous infusion of ACTH 250 micrograms. Seven E.H. patients were similarly studied in the condition of low and high sodium intake and plasma renin activity (PRA) was also measured at the same time. P.A. showed significantly higher ALD than that of E.H., whereas DHA-f and DHA-s levels of these patients were lower than that of E.H. In men with E.H., TEST level showed step down response to ACTH. Low sodium intake induced a marked increase of ALD and PRA in E.H. In low sodium intake other mineralocorticoids such as DOC and B showed high value at 60 min (20% t value) after ACTh infusion. From these observations, it was suggested that the mineralocorticoids have a good response to ACTh in low sodium intake rather than high sodium intake, especially ALD has a clearly augmented response to ACTh. On the other hand, PRA showed only a reserve response to ACTH infusion. PMID- 6275462 TI - [Contrast radiographic study of ductal pathology of the breast]. AB - On the basis of personal experience, the possibility and the limits of galactography are discussed. Galactography has been used by the authors in any cases of discharge from the nipple, and has shown to be the only radiological examination able to discover intraductal tumours. Furthermore it allows to establish the site and extension of the lesions. Among the limits of the method the impossibility to distinguish between benign and malignant lesions is stressed out. PMID- 6275464 TI - [Neuromuscular transmission troubles in a case of acute lithium intoxication (author's transl)]. AB - In the course of an acute intoxication by lithium, neuromuscular junction has been tested by 1 sec bursts of repetitive stimulation. Recorded data concern: (1) inhibition of acetylcholine synthesis, at the time of high seric rates; (2) exhibition, at rather lower concentrations, of two types of potentiation: a primary, moderate potentiation, high frequency-sensitive (30 or 50 HZ), and a more lasting cumulative potentiation, enhancing the post-tetanic potentiation by nearly 300%. Results are compared with some of the neurophysiological data in the literature. PMID- 6275463 TI - [Parameters of a cell population synchronized for radiosensitivity studies (author's transl)]. AB - Human cultured cells (EUE) were synchronized by the method of the mitotic harvest and the degree of synchronization, during the first duplication interval, determined by various tests which include growth curves, mitotic index cell-size distributions. Values of synchronization index initially greater than 90% and desynchronization rate of about 2%/hour were evaluated. The survival after 1.75 Gy of 31 MeV protons irradiation shows a sensitive period in the late G1 followed by an increase in radioresistance to a maximum in S. PMID- 6275465 TI - [Purine catabolism in rat brain (author's transl)]. AB - In soluble rat brain fraction, the specific activities of purine nucleoside phosphorylase, guanine deaminase, 5'Nucleotidase and adenosine deaminase, decrease in their mentioned order. A kinetic parameter comparison between these enzymes shows that 5'Nucleotidase with AMP has the lowest KM and the greatest Vmax values, while purine nucleoside phosphorylase has its lowest KM and its greatest Vmax values with guanosine and with inosine, respectively. The enzymes activity is not modified by the metabolic intermediates differently from their own reaction products which behave as competitive inhibitors. PMID- 6275466 TI - Effect of physiological electron donors and acceptors on F1-ATPase. AB - The hydrolytic activity of F1-ATPase isolated from rat liver was enhanced in the presence of NADH, FADH2, QH2 or reduced cyt c. The extent of this activation depended largely on substrate concentration. F1-ATPase sensitivity to bicarbonate or dinitrophenol activators decreased in the presence of any of those electron donors, which originated as well a slight sensitivity to oligomycin and a sensitivity increase to the inhibitory anion OCN-. In the presence of oxidized carriers the sensitivity to bicarbonate, dinitrophenol, or OCN- was not modified, and the enzyme remained oligomycin insensitive. PMID- 6275467 TI - Insulin receptor interaction in human placental plasma membranes. AB - Insulin-receptor interaction in partially purified preparations of human placental plasma membranes from normal mothers at term of pregnancy has been characterized. 125I-insulin became rapidly and reversibly bound to plasma membranes, being time and temperature dependent. The binding readily appeared at 1.0 ng/ml insulin concentration which falls within the physiological range of peripheral blood. Low levels of unlabeled insulin inhibited binding; 20 ng/ml insulin produced fifty per cent inhibition. Scatchard plots of data from competitive insulin binding proved to be curvilinear. The insulin greater ability for binding observed in this preparation can be explained by the purification degree achieved at the plasma membranes. 125I-insulin was less degraded by partially purified placental plasma membranes than by a microsomal-membrane preparation obtained without differential centrifugation in sucrose linear gradient. All these properties strongly suggest that the insulin-binding sites characterized in the plasma membrane fraction of the placenta represent biologically important receptors to hormone. PMID- 6275468 TI - [Cerebral irradiation as adjuvant therapy in the treatment of small cell carcinoma of the lung (author's transl)]. AB - The authors present the results of a non randomized study of 33 patients suffering from small cell carcinoma of the lung. After a similar course of monthly chemo-therapy (Endoxan Oncovin, Natulan, CCNU) and mediastinal irradiation in a dose of 30 grays in 10 fractions over two weeks, 14 patients were given adjuvant cerebral irradiation, the 19 others made up the control group (30 grays to the brain overall in 10 fractions over two weeks). With a minimum follow up for 12 months, 42% of the control group had evidence of cerebral secondaries, as opposed to none in the irradiated group with any neurological signs whatsoever. The indisputable efficacy of adjuvant cerebral irradiation contrasts with the absence of any significant improvement in survival time : 2 patients are alive and in complete remission, 1 in each group, while the mean survival was 12.3 and 11.7 months respectively. This is explained by the continued occurrence of extra-cerebral metastatic deposits such as the liver or mediastinum where the efficacy of systemic therapy remains uncertain. The therapeutic approach is currently orientated in two directions : - to raise the dose delivered to the mediastinum even if it appears radiologically normal. - to continue research into new chemo-therapeutic combinations more specifically active at the hepatic level. PMID- 6275469 TI - Human basement membrane antigens from lung, placenta and kidney. AB - Three human basement membranes, glomerular basement membrane (GBM) from renal cortex, alveolar basement membrane (ABM) from lung parenchyma and trophoblast basement membrane (TBM) from the terminal villi of placenta have been isolated by sieving and sonication techniques. Canine GBM and ABM were also prepared. There were marked differences among the membranes from human tissues. Compared to GBM, TBM had very little collagen but contained high concentrations of charged amino acids. ABM was intermediate in composition between GBM and TBM and contained desmosine and isodesmosine indicative of the presence of elastin. Canine ABM (c ABM) did not contain desmosine or isodesmosine. In the canine system an antigen was detected in ABM which was not present in GBM. The membrane preparations were analyzed for fibronectin content using a specific antiserum to fibronectin. This glycoprotein could not be detected in GBM whereas it was present in ABM in amounts up to 0.8% and in TBM in amounts as high as 7.2%. All the membranes induced the formation of precipitating antibodies in rabbits. Soluble material obtained from the membranes by alkali extraction, reduction of disulfide bonds, enzymatic digestion with elastase, plasmin or collagenase provided immunologically reactive fragments. These soluble fragments gave reactions of identity among the three basement membranes in immunodiffusion reactions in gels with antisera raised to all three BMs. The finding that plasmin digests basement membranes suggests that it may play a role in connective tissue remodeling. The fact that elastase degrades basement membranes provides an endogenous system for injury which may be triggered by infections. PMID- 6275470 TI - Isolation of human glomerular basement membrane antigens by affinity chromatography utilising Goodpasture's kidney antibody eluates. AB - The soluble material produced from a 96-hour digest of sonicated human glomerular basement membranes with purified collagenase was shown to contain at least 12 components by sodium dodecyl sulphate-polyacrylamide gel electrophoresis with molecular weights ranging from 20 x 10(3) to greater than 200 x 10(3) daltons. Five of these components produced antigenic peaks after two-dimensional immunoelectrophoresis into agar plates containing rabbit antiserum to the solubilised glomerular basement membrane. Two groups of antigenic components were demonstrated in these gels by two-dimensional immunoelectrophoresis autoradiographic techniques utilising 125I-labelled collagenase-digested human glomerular basement membranes reacted with antibodies eluted at acid pH from the kidney of the Goodpasture's patient. This acid eluate was shown to contain contaminants of alpha 1-antitrypsin and albumin co-eluting with the antibodies bound to the glomerular basement membrane. After removal of these contaminants, the antibodies were linked to cyanogen bromide-activated Sepharose and used to affinity purify four antigenic fractions from the collagenase-digested glomerular basement membrane. These fractions were eluted by 0.2 M glycine pH 2.8 and with 0.5 M ammonium thiocyanate and had molecular weights of 22-25, 43-45, 65-70 and 205 x 10(3) daltons. The smaller molecular weight components were shown to be common to both included and excluded fractions obtained by Ultragel AcA 44 chromatography of the digested material in 10 mM phosphate pH 8. This suggests that the larger molecular weight component would be an aggregate of a smaller component. Preliminary carbohydrate and amino acid analyses indicated that the different elution procedures elicited antigens with different chemical characteristics. PMID- 6275471 TI - EC collagen: biosynthesis by corneal endothelial cells and separation from type IV without pepsin treatment or denaturation. AB - Primary cultures of rabbit corneal endothelial cells were labeled with proline and the medium proteins were fractionated by sedimentation through sucrose density gradients. A new collagen, endothelial cell (EC) collagen, was completely separated from type IV collagen without denaturation by this technique. Sodium dodecyl sulfate electrophoresis demonstrated EC collagen fragments of 180K, 120K, and 60K daltons. Prior pepsin treatment produced only the 60K fragment after electrophoresis. The cyanogen bromide peptide pattern of the 180K EC peptide demonstrated that EC was different from type IV. A preliminary structural model for the largest helical form (approximately 540K daltons) of EC collagen is presented. PMID- 6275472 TI - Morphiceptin ( beta-casomorphin (1-4) amide): a peptide opioid antagonist in the field stimulated rat vas deferens. AB - The effect of Morphiceptin (H-Tyr-Pro-Phe-Pro-NH2) on opioid inhibition of field stimulated rat vas deferens (RVD) was studied. The morphiceptin was found to produce a weak inhibition of the twitch response of the RVD. In addition morphiceptin antagonized full agonists acting on the same receptor system (i.e. beta-endorphin selective antagonist for the delta receptor system of RVD by virtue of being a weak partial agonist. PMID- 6275473 TI - 3-Chlorodibenzazepine binding to cholinergic receptors and ion channels. AB - The interactions of 3-chlorodihydrodibenzazepines with nicotinic receptors and ion channels from Torpedo californica electric organ and muscarinic receptors from rat brain were revealed by their inhibition of the binding of specific radiolabelled probes. Desmethylchlorimipramine (DCI) was twice as active as chlorimipramine (CI) in inhibiting [3H]phencyclidine and [3H]perhydrohistrionicotoxin binding to nicotinic ion channels; inhibition constants were 0.13 and 0.28 micro M in the presence and 1.2 and 3.4 micro M in the absence of carbamylcholine. Both of the drugs had marginal affinity for the nicotinic receptor (i.e., the [3H]3-quinuclidinyl benzilate binding to muscarinic receptors from rat brain stem and cerebral cortex with inhibitions constants below 0.1 micro M. Little regional specificity or activation by N-ethylmaleimide was apparent in the muscarinic binding of CI and DCI, properties associated with the binding of receptor antagonists. PMID- 6275475 TI - Effect of infection of the bursa of Fabricius of day-old chickens with infectious laryngotracheitis virus on subsequent bursa development and antibody responses. PMID- 6275474 TI - Adrenal Size and function after irradiation with 131I-6 beta-Iodocholesterol in rabbits. AB - Administration of high doses of 131I-6 beta-iodocholesterol to rabbits is followed by marked size reduction of the adrenal glands with concomitant changes of the histological structure and with a decrease of serum cortisol. Stimulation of 131I-6 beta-iodocholesterol uptake into the adrenals by additional administration of ACTH had only a minor additional effect on adrenal weight. However, with further accumulation of labeled cholesterol in the adrenals the radioactivity was reduced in blood urine, and the ovaries. PMID- 6275476 TI - Some effects of copper deficiency on leucocyte function in sheep and cattle. AB - Peripheral blood leucocytes from copper-deficient (less than 8 mumol per litre plasma) and copper-sufficient (more than 8 mumol per litre) ewes and lambs were tested for their ability to ingest and kill Candida albicans in vitro. Killing capacity was significantly reduced in both copper-deficient ewes and lambs, although phagocytic capacity was apparently unimpaired. Similar results were obtained with copper-deficient calves, in which it was further shown that copper repletion restored candidacidal activity. Leucocyte and erythrocyte superoxide dismutase (LSOD and ESOD respectively) activities were both significantly decreased in hypocupraemic ewes and there was a linear relationship between their activities in the two cell types: LSOD activities were reduced by approximately 35 per cent and ESOD activities by 67 per cent. Cyanide inhibition tests indicated that the manganese-dependent form of the enzyme contributed little to the total activity of LSOD or ESOD. There was a tendency for leucocytes from copper-deficient ewes to release greater amounts of superoxide anion after stimulation with opsonised zymosan. Their viability was similar to that of leucocytes from control sheep and was unaffected for up to three hours after stimulation. PMID- 6275477 TI - Cell mediated and humoral immune response of chickens to live infectious bronchitis vaccines. AB - A lymphocyte transformation microassay was used to study cell-mediated immune (CMI) responses in chickens following inoculation with live infectious bronchitis virus vaccines H 120 and H 52 and challenge with Massachusetts-41 (M-41). Humoral immunity was monitored for comparison, using haemagglutination inhibition (HI) microassay. Specific stimulation of infectious bronchitis virus sensitised lymphocytes was demonstrated by their lack of response to Mycoplasma synoviae and Newcastle disease virus antigens. Positive infectious bronchitis virus stimulation indices appeared before HI titres and ranged from 2.0 to 7.1 over a period of five to 33 days after primary vaccination with H 120. Revaccination with H 52 produced a strong secondary HI response and a relatively weak CMI response. Challenge of vaccinated and revaccinated birds also resulted in strong HI and weaker CMI secondary responses but the more pronounced CMI responses were in the revaccinated-challenged group. There was no correlation between CMI and HI antibody production but a negative correlation was demonstrated between peak CMI responses after vaccination and severity of clinical symptoms after subsequent challenge. PMID- 6275478 TI - Pathology of natural canine adenovirus pneumonia. PMID- 6275479 TI - Preparation and efficacy of an inactivated subunit vaccine against Aujeszky's disease virus infection. PMID- 6275480 TI - [Rotavirus and acute diarrhea in infancy: comparison of hospitalized and outpatient cases (author's transl)]. PMID- 6275481 TI - [Progressive centripetal degeneration in polyneuropathies (author's transl)]. AB - This is a clinicopathologic report on three patients with sensory polyneuropathies of different origin. Sensory loss involved all four limbs reaching the upper third of the thighs and the elbow level or higher, in all three patients. In addition to the limbs the central region of the anterior aspect of the trunk, from lower abdomen up to level T2, and on the top of the scalp were involved. There was minimal weakness. This pattern of sensory deficit can best be explained by a length dependent degeneration of fibers. Familial amyloidosis, Portugese type, was responsible for the neuropathy in the first patient, diabetes mellitus in the second and alcoholism in the third one. On teased nerve fiber study, single regenerating fibers were isolated on sural nerve biopsy specimens from patients 1 and 2. Segmental demyelination and/or remyelination occurred in 11 per cent of the fibres in patient 1, in 36 per cent in patient 2 and in 4 of the 19 fibres isolated in patient 3. On cross sections of nerve specimens embedded in Epon there was a striking loss of myelinated fibres which was less important and predominated on smaller fibres in patients 1 and 2. On electron microscopic examination loss of unmyelinated fibres was conspicuous in all three patients. On single fiber studies as well as on sections of embedded specimens, myelinated fibres occasionally showed demyelination in contact to amyloid deposits. The present study demonstrates that in this pattern of neuropathy degeneration of myelinated fibers begins in the distal part of longest axons and may be associated with axonal sprouting in more proximal parts of degenerating axons. As the neuropathy progresses axons of shorter and shorter length become involved. PMID- 6275482 TI - [Thecomas and granulosa cell tumors: clinical, morphological and therapeutic aspects]. PMID- 6275484 TI - EBV infections in brazil. III -- Infections mononucleosis. PMID- 6275483 TI - [Diarrheic syndrome and beta cell tumors of the pancreas: the Verner-Morrison syndrome: report of a case]. PMID- 6275486 TI - [Current data on hormonal receptors in breast cancer]. PMID- 6275485 TI - [What is new in the field of viruses in hepatology?]. PMID- 6275487 TI - [Pharmacology of converting enzyme inhibitors (captopril and related products)]. PMID- 6275488 TI - [Current clinical use of converting enzyme inhibitors]. PMID- 6275489 TI - [Rational use of curariform drugs in anesthesia and resuscitation]. PMID- 6275490 TI - [Inhibitory action of magnesium thermophosphate on the fecundity of planorbids and its possible significance in the control of schistosomiasis mansoni]. PMID- 6275491 TI - [Seroepidemiology of Epstein-Barr virus]. PMID- 6275492 TI - Effects of pentoxifylline and various methylxanthine derivatives on transepithelial water permeability of frog urinary bladder. AB - Theophylline, pentoxifylline and MIX (isobutyl methylxanthine) increase the water permeability of frog urinary bladder when added to the serosal side of the epithelium; this hydrosmotic effect is explained by an inhibition of adenylate cyclase. Both pentoxifylline and MIX exhibit an higher biological activity than theophylline. In contrast to theophylline, pentoxifylline and MIX stimulate the water permeability when added to the mucosal side of the epithelium; this probably results from a better penetration of these drugs into cells. PMID- 6275493 TI - Adenylate cyclase activity in gastric mucosal biopsies and cAMP in gastric juice before and after cimetidine treatment in healthy subjects. AB - Previous studies have revealed augmented parietal call sensitivity to histamine stimulation after cessation of cimetidine treatment in healthy subjects. To elucidate one possible mechanism for this effect, we measured cAMP in gastric mucosa and gastric juice during histamine stimulation in vivo and histamine stimulated adenylate cyclase activity in vitro in biopsy material obtained by gastroscopy. In eight healthy subjects the cAMP content of gastric corpus mucosal biopsies was measured before, during, and after infusion of 2.5 and 25.6 microgram . kg-1 . h-1 of histamine dihydrochloride. No increase in cAMP content was found. In 10 healthy subjects cAMP secretion into gastric juice was studied before and 36 to 84 h after cessation of 4 weeks' treatment with cimetidine, 1 g/day. No significant increase of cAMP output was found in response to infusion of 2.5 and 25.6 microgram . kg-1 . h-1 of histamine dihydrochloride either before or after the treatment period. Thus histamine does not stimulate cAMP secretion into gastric juice in man. In the same 10 subjects histamine-stimulated adenylate cyclase activity was studied in vitro in biopsy material obtained before and 60 h after cessation of cimetidine treatment. The dose-response curve of the adenylate cyclase activity was not shifted to the left. The results indicate that the augmented parietal cell sensitivity to histamine, seen after stopping cimetidine treatment in healthy subjects, is not due to increased sensitivity of the adenylate cyclase-cAMP system to histamine stimulation. PMID- 6275494 TI - Adrenergic modulation of the release of pancreatic polypeptide after intraduodenal and oral glucose in man. AB - Intraduodenal infusion of glucose increased the concentration of pancreatic polypeptide (PP) in serum from 13.4 (4.0-20.4) to 36.9 (20.7-81.3) pM. Alpha blockade with phentolamine increased the PP concentration from 15.0 (4.0-23.7) to 24.9 (4.6-50.2) pM, and after intraduodenal glucose to 46.8 (23.6-132.8) pM. The PP release after intraduodenal glucose was small, transient, and significantly reduced when beta blockade with propranolol was administered. Oral glucose increased the concentration of PP from 19.3 (4.2-37.0) to 61.1 (14.1-141.7) pM. Isoprenaline increased the PP concentration from 13.5 (4.6-33.8) to 56.0 (5.7 137.3) pM, and after oral glucose to 77.5 (25.3-134.7) pM. The increase in PP concentration was eliminated when propranolol was added to isoprenaline. We conclude that an intestinal phase of PP release exists after intraduodenal glucose in healthy humans, and that the PP release after intraduodenal and oral glucose can be modified by the adrenergic nervous system. Alpha blockade stimulates the PP cell; beta blockade or stimulation respectively inhibits or stimulates the PP cell. PMID- 6275495 TI - A prospective, partly randomized study of the effectiveness of repeated examination of the colon after polypectomy and radical surgery for cancer. AB - Results of 629 colonoscopies and 130 double-contrast examinations, performed during the first 2 years of a prospective, partly randomized cancer prophylactic programme, are reported. The patients were not more than 75 years old and had no previous diagnosis of adenoma or cancer. Repeated colonoscopy 3 months after polypectomy in 80 patients increased the chance of obtaining a complete colonoscopy from 70% to 86%, and the hepatic flexure was reached in a further 9%. Thirteen had polypectomy during the second colonoscopy. The gain in number of patients with adenomas was highest in those with sessile villous adenomas, carcinoma in situ, and mucosal carcinoma. Repeated colorectal examinations every 6 months in these resulted in recurrence rates of 13 of 33, 3 of 23, and 2 of 15 (18 months), respectively. The same figures for patients with stalked polyps, allocated to examination every 6 months, were 3 of 34, 1 of 21, and 2 of 14. Colonoscopy was also performed within 3 months of radical surgery for colorectal cancer, and 35 of 142 patients had polypectomy. Repeated examinations every 6 months resulted in recurrence rates for adenomas of 11 of 85, 3 of 46, and 1 of 34. One new cancer after 12 months could be treated radically. Five uncomplicated laparotomies were performed after the 629 colonoscopies because of perforation or bleeding. PMID- 6275496 TI - High resolution SEM of cultured cells: preparatory procedures. PMID- 6275497 TI - Morphology of temperature-sensitive strains of viral-transformed 3T3 mouse fibroblasts in vitro as determined by SEM and STEM. AB - The morphological characteristics of BALB/c 3T3 mouse fibroblasts transformed with two temperature-sensitive strains of simian virus 40 (SV-40) were studied. When grown at the permissive temperature (32 degrees C, transformed phenotype) strains A7B4b and A209B4a both closely resemble the morphology described for wild type SV-40 cells. Scanning electron images show numerous knobs and filopodia as characteristic surface features in both subconfluent and confluent cultures. Scanning-transmission electron images reveal that the two strains are also similar in intracellular structure. When grown at the restrictive temperature (39 degrees C, untransformed phenotype) the two strains differ from each other. The A209B4a strain is more elongated in subconfluent cultures, and shows interactions of filopodia at cell-cell interfaces when confluent similar to those described for wild-type 3T3 cells. This strain also shows intracellular accumulation of polymorphic material thought to be secondary lysosomes. The A7B4b cells are flattened but generally not elongated when subconfluent, and do not show any obvious interactions at confluency. Intracellularly, this strain also differs from A209B4a. PMID- 6275498 TI - Long-term effects of endocrine treatment on serum pituitary hormones in advanced prostatic carcinoma patients. AB - Peripheral serum concentrations of FSH, LH, prolactin and ACTH were measured in 22 patients with advanced prostatic carcinoma treated by castration, polyestradiol phosphate (Estradurin) administration and a combination of castration and Estradurin administration during the first 12 months of treatment. Estradurin treatment alone (80 mg i.m.l once a month) did not result in any significant changes in the circulating concentrations of FSH, LH and prolactin. Therefore, the clear-cut inhibition of testicular steroidogenesis observed under this kind of treatment does not appear to be due to an inhibition of pituitary gonadotropin secretion, and is most likely due to a direct estrogen effect on Leydig cells. Castration led to grossly elevated serum FSH and LH levels. In this group, serum FSH remained at a high level, whereas LH was close to pretreatment levels 9 months after castration, suggesting differences in pituitary capacity to secrete FSH and LH under these conditions. Concentrations of circulating FSH, and to a lesser extent LH, in the combination treatment group were between those found in the castration and estrogen-only treatment groups, suggesting that the secretion of both gonadotropins can be suppressed by estrogen. No changes in serum prolactin and ACTH concentrations were seen in the three treatment groups. PMID- 6275500 TI - [Herpesvirus infections in puppies: virological data and histopathological findings]. PMID- 6275499 TI - Arthralgia and crystal deposits in Crohn's disease. AB - Ten patients with Crohn's disease and recurrent pain in the knee joints were subjected to arthroscopy. Biopsies obtained from the synovial membrane were examined under polarizing light microscopy. The arthroscopy revealed crystalline deposits in 7 patients and the microscopic examination of the synovial membrane demonstrated positively birefringent crystals in 4 patients. The crystals with positive birefringence had the rod or rhomboid shape typical of pyrophosphate crystals. As arthroscopy crystals in 7 patients and polarizing microscopy revealed crystals in one further patient, crystal deposits were thus found in 8 patients altogether. All patients had normal serum uric acid values. The crystal deposits were interpreted as pyrophosphate and their possible connection with the recurrent arthralgia in Crohn's disease is discussed. PMID- 6275501 TI - [Involvement of the spine in articular chondrocalcinosis]. PMID- 6275503 TI - [Dislocation of the hip during the first year. Requirements for prevention]. PMID- 6275502 TI - [How to manage an inflammatory syndrome]. PMID- 6275504 TI - [Anti-inflammatory agents and rheumatic diseases. New products: why make them?]. PMID- 6275505 TI - [Diagnosis and therapy of early-onset osteoporosis]. PMID- 6275506 TI - [Schwartz-Bartter syndrome]. PMID- 6275507 TI - [Profile of blood pressure in exertion]. PMID- 6275508 TI - [Hypertension crisis: diagnosis and emergency therapy]. PMID- 6275509 TI - [Myocardial scintigraphy under stress in the detection of coronary insufficiency]. PMID- 6275510 TI - [New mode of administration for an anti-coagulant]. PMID- 6275511 TI - [Weather conditions in myocardial infarction, cerebrovascular strokes and suicide attempts]. PMID- 6275512 TI - [Evaluation in the arteritis patient, of the peripheral and cardiac effects of vasodilator agents by telethermography, thallium 201 scintigraphy and measurement of ejection fraction]. PMID- 6275513 TI - [Must large tonsils be removed in children?]. PMID- 6275514 TI - [Inhibition and its manifestations]. PMID- 6275515 TI - [Legionnaires' disease]. PMID- 6275516 TI - [Bacterial antigen therapy in asthma]. PMID- 6275517 TI - [Cystitis]. PMID- 6275518 TI - [Diagnosis of peripheral adenopathy in children]. PMID- 6275519 TI - [Abscess of the brain in 1981]. PMID- 6275521 TI - [False migraines]. PMID- 6275520 TI - [Principal aspects of depression and their therapy]. PMID- 6275522 TI - [Echographic surveillance of induction of ovulation]. PMID- 6275523 TI - [Human basophil degranulation test (HBDT) for diagnosis of respiratory allergic disorders. A study of eighty-four HDBT performed in forty-seven patients (author's transl)]. AB - The HBDT uses basophils of patients with respiratory allergic disorders. In this simple method the basophils are stained by toluidine blue and the erythrocytes are destroyed. The basophils are then exposed to solutions containing different concentrations of the antigen. The basophil degranulation rate in response to a given antigen can be determined. The test is positive if the degranulation rate is over 50% and negative if the rate is under 30%. Our clinical study induces 47 patients. The correlate rate between HBDT and skin tests is 81,5%. This test's technical simplicity allows it's routine use by a non specialized laboratory. PMID- 6275524 TI - [Still's disease in the adult (author's transl)]. AB - Four new cases of adult Still's disease have been observed in the past three years. Clinical and biological features are high fever, polyarthralgia, macular rash on forearm, pericarditis, splenomegaly, lymphadenopathy, alopecia, anemia, hyper-leucocytosis, raised ESR. Transient neurological findings may also be observed with abnormal reflexes, cranial nerve paralysis. High dose steroids treatment can be used in case of systemic involvement. The long term prognosis is usually good with treatment. In conclusion, the diagnosis of Still's disease is essentially clinical and should not be mistaken for septicemia. PMID- 6275525 TI - [Surgery for bronchogenic carcinoma in patients over seventy years old (author's transl)]. AB - 125 patients aged over 70 years underwent operation for bronchogenic carcinoma at the Centre Chirurgical Marie Lannelongue for a period extending from 1963 to 1976. There were 123 male and 2 female patients. 112 had a squamous cell carcinoma. A total of 32 patients had a pneumonectomy performed, 86 a partial resection (lobectomy or bilobectomy) and 7 an exploratory thoracotomy. Post operative death was definitely higher (15,6%) in the pneumonectomy group than in the lobectomy one (2,6%). The main cause of post-operative death (62%) was pulmonary embolism. Post-operative complications were mainly related to cardiac arrhythmias. Overall survival rate after 5 years was 25,2%, slightly higher in the lobectomy group (25,9%) than in the pneumonectomy group (22,2%), higher in the differentiated squamous cell carcinoma group (31,1%) than in the undifferentiated (6,3%). The quality of life was reasonably satisfactory in most of the patients considering their age. Surgery for bronchogenic carcinoma in patients over 70 years old is recommended specially with limited resection is possible. PMID- 6275526 TI - [Changes in blood cell counts and coagulation factors during labor and after delivery (author's transl)]. AB - Hemostasis and blood cell counts were studied during pregnancy, during labor, and after delivery in 76 women. Results were compared with data from a control group of 24 women. Previously published data are reviewed and the variations of studied factors are discussed. No reliable biologic test is available at present for assessing the risk of puerperal thromboembolism. PMID- 6275527 TI - [Surgery in patients with chronic renal failure on maintenance dialysis (author's transl)]. AB - Among 606 patients on maintenance dialysis in the same unit, 111 (18.3%) underwent various surgical procedures. 14 additional patients on dialysis in other units were included in this group. A total of 138 surgical procedures were performed in these 125 patients, of which 112 were on hemodialysis. In 57 patients, surgery was warranted by a complication directly related to the chronic renal failure. In 81 patients, surgery was indicated because of an associated condition, a complication resulting from therapy, or a coincidental surgical affection. After these 138 procedures, 27 post-operative complications (19,6 %) and 15 deaths (10,86 %) were reported. The mean age for deceased patients is 66 years. These findings show that surgical morbidity and mortality, even after major procedures, are not prohibitive in patients on dialysis, if certain measures are taken during dialysis, anesthesia, surgery, and post-operative care. These measures are reviewed in the study. PMID- 6275528 TI - [Venous thrombosis, circulating anticoagulant and systemic lupus erythematosus. Two cases reported in two identical HLA sisters (author's transl)]. AB - The observation of two cases of thrombophlebitis of the calf which occurred in two sisters within a period of three years is reported. The investigation revealed in both cases the presence of a circulating anticoagulant with anti prothrombinase activity in association with biological signs of systemic lupus erythematosus (L.E.): anti-DNA antibodies, decreased complement levels, false positive BW reactions. In one patient, the skin biopsy showed a continuous IgM band on immuno-fluorescence. After respectively 4 years and 18 months follow-up both patients are still free from clinical symptoms. A review of the literature is presented and the clinical and etiological significance of the presence of a circulating anticoagulant, its relationship with L.E., particularly with "latent" L.E. (presence of biological signs only), and its association with thrombophlebitis are discussed. PMID- 6275530 TI - [Mechanisms of acute pulmonary edema during meningococcal shock in children (author's transl)]. AB - Meningococcic shocks may induce fatal acute pulmonary edemas. Ten children, who died in this condition, have been compared to 19 others, who during the treatment of the shock, developed right heart failure, without acute pulmonary edema (8 deaths, 11 survivals). The three groups of children could not be differentiated regarding their clinical status on admission, during the shock phase, or the treatments administered. Composition, volume and speed of administration of fluids were similar in the three groups. An increase in pulmonary capillary permeability could have occurred, and lead to the development of acute pulmonary failure, as presented by the children of the first group. PMID- 6275529 TI - [Osteosclerotic myeloma. Report on three cases and review of the literature (author's transl)]. AB - Three cases of osteosclerotic myeloma are reported. Clues for diagnosis are agarose gel electrophoresis and sclerotic bone biopsy. Review of 50 cases of literature points out frequency of peripheral neuropathy and Ig A type Immunoglobulin. Iliac crest biopsy shows thickening of bone structures and plasmocytosis in medullary spaces. Main X-Ray signs are mentioned and pathogenesis is discussed. PMID- 6275531 TI - [Long-term result (5-10 years) of radiotherapy of inoperable carcinomas of the prostate (author's transl)]. AB - A series of 59 consecutive patients with locally inoperable adenocarcinomas of the prostate were irradiated following a strict protocol and controlled dosimetry in a national cooperative study in the USA. Twenty-one of these patients are living without signs of recurrence or recurrence or metastases. Additional patients who lived 5 to 10 years after treatment without recurrence or metastases died of other causes. Autopsies in several of these cases failed to show residual or metastatic tumor. The failures are primarily due to bone metastases outside of the irradiated territory. The urethral and rectal complications are rare; they can be reduced by further fractionation. PMID- 6275532 TI - [Tiapride in detoxication of chronic alcoholics (author's transl)]. AB - Tiapride was used in 55 chronic alcoholics. It has a sedative effect on the anxiety, aggressiveness and agitation observed during the alcohol withdrawal syndrome. It is also effective against tremor, insomnia and fatigue. Fatigue or depression do not occur as side-effects. Tiapride induces a psychological feeling of wellbeing which is heightened by continuation of detoxication and general management. PMID- 6275533 TI - [Surgically curable hypertension (author's transl)]. AB - A case of hypertension was cured by simultaneous surgical treatment of an obstructive urolithiasis associated with a pheochromocytoma. Primary devascularization of the adrenal tumor, reducing blood pressure and cardiac rhythm variations was allowed by preoperative arteriography. PMID- 6275534 TI - [Gastric carcinoma. Therapy and prognosis. A report of a surgical series of 235 cases (author's transl)]. AB - In a series of 235 gastric cancer cases, 222 of them justified in doing an operation and 155 an excision. The operation mortality rate fluctuated between 25 p. cent and 10 p. cent according to the type of treatment. For the whole group a five years actuarial survival was 13,3 p. cent; 22,5 p. cent in the case of excision with the aim of curing. A study of prognosis factors brings out the favourable role played by tumors of the ulcerated type, not reaching the serous wall, without lymph embole in the wall, and of course without metastases or extension at section cuts. PMID- 6275535 TI - [Primary sclerosing cholangitis (author's transl)]. AB - Two case of Primary Sclerosing Cholangitis (PSC) associated with inflammatory bowel disease are reported. In one of these cases, inflammatory bowel disease was asymptomatic and discovered only by performing colonoscopy. Endoscopic retrograde cholangiography is of particular value, showing the specific lesions of PSC. Treatment is only symptomatic. PMID- 6275536 TI - [Amyloidosis and polycythemia vera (author's transl)]. AB - The authors report a case of systemic amyloidosis with predominantly hepatic, splenic and renal deposits and polycythemia vera with secondary myelofibrosis. The course of both diseases during a 9-year-follow-up and the late appearance of a monoclonal IgM lead the authors to examine a possible relationship between the two diseases. PMID- 6275537 TI - [The lung after radiation therapy. Report of a case of primary lymphosarcoma of the lung (author's transl)]. PMID- 6275538 TI - [Pheochromocytoma in black African. Five cases whose three ectopic. Review of literature: thirty-three cases (author's transl)]. AB - The authors report five cases of pheochromocytoma observed in black African people, followed in the Hospital University Center of Treichville. Three of these pheochromocytoma were in ectopic location. In connection with these observations, the authors review the African literature, regrouping 33 cases, which allows to define the principal aspects of the disease in black African patients. The over morbidity amongst the female patient, the frequent discovery during pregnancy or even at the onset of labour, the frequency ectopic localization are quite original characteristics. PMID- 6275539 TI - [Prophylaxis with cefazolin in surgery of the hip (author's transl)]. AB - Our study shows that prophylactic use of an adequate antibiotic appears to be warranted in osteoarticular surgery, during the immediate pre and post-operative periods. The mechanism by which this prophylaxis is effective remains under debate. Surgeons should be encouraged to abandon the use of wide-spectrum antibiotics or combinations of several antibiotics for prophylactic purposes. The only useful prophylaxis is the prescription of one antibiotic effective against the most common agent i.e. staphylococcus aureus. Our results suggest an early action of the antibiotic, as soon as the leukocytes and microorganisms make contact. Selection of highly resistant microorganisms by polyvalent antibiotic therapy is precluded by the prophylactic regimen advocated here. Use of cefazolin during a very short period appears to be a satisfactory solution to prophylactic problems in osteoarticular surgery. PMID- 6275540 TI - [Hematotoxicity of thiamphenicol. Report of two cases of acute erythroblastopenia (author's transl)]. AB - Thiamphenicol is known to produce two hematological side effects: a decrease in blood cell count concerning mainly red blood cells; in this case regression is observed after withdrawal of the drug; the frequency of this side effect appears to be higher than that observed after chloramphenicol administration. Severe and sometimes fatal pancytopenia is rare and related to primary bone marrow failure; its incidence seems to be increased by high dosages, long-term or repeated administration, interaction with other hematotoxic drug administration, renal failure, elderly or immune suppression. PMID- 6275541 TI - [Giant-cell arteritis with initial resistance to corticosteroids (author's transl)]. AB - Treatment of giant-cell arteritis rests on corticosteroids. Three observations of Horton arteritis with initial resistance to cortisone are reported. Diagnosis was substantiated by the examination of biopsy specimens. Resistance to therapy was overcome by changing to another corticosteroid. Observations such as those reported here are very uncommon. PMID- 6275542 TI - ["Lymphoid" blastic transformation in chronic myelogenous leukemia. Report of three cases (author's transl)]. AB - Three cases of chronic myelogenous leukemia (CML) were studied, occurring in 22, 43, and 30-year-old-men. Two observations (nos. 1 and 3) concerned typical CML, treated by discontinuous busulfan; in the last patient (no. 2), also presenting in addition with a constitutional deficiency from Hageman factor, diagnosis (Ph 1 chromosome) was based on a moderate leukocytosis with myelemia, spontaneously regressive for more than one year. Chronic phase duration was 17, 16 and 8 months respectively. During the first blast crisis, abnormal cells were rather of granular type in one case (no 1), undifferentiated in the other two. In the three observations, complete remission was easily obtained with prednisolone - vincristine but revealed very brief; 2, 2 and 4 months. Among the three patients a second blast crisis was preceded, in two cases (nos. 1 et 3), by a new CML phase during 3 and 1 months respectively. Treatment was then purely palliative by 6-mercaptopurine and hydroxyurea. PMID- 6275543 TI - [Avulsion of the ureter due to external violence. Report of four cases (author's transl)]. AB - Four cases of traumatic ureteral damage are presented. The etiology of ureteral injuries is gunshot wounds for one case and motor vehicle accidents for three cases. Need for a high index of suspicion and diagnostic measures with IUP is stressed. Prompt operative management is recommended. Conservative surgery is emphasized. PMID- 6275544 TI - [Three cases of regressive hypertension after renal trauma. (author's transl)]. AB - Three cases of post-renal contusion hypertension are reported. The treatment was conservative in the three cases, and we think it had no incidence on return to normal arterial pressure. There was a good correlation between arterial pressure and renal vein renin activity. PMID- 6275545 TI - [A clinical study of tiapride in ten patients with chronic respiratory failure and in ten patients before fiberoptic bronchoscopy (author's transl)]. AB - Ten patients with chronic respiratory failure (PaO2 = 56,19 +/- 10,54; PaCO2 = 49,17 +/- 7,31 torr) were given intramuscular injections of tiapride, Changes in alveolar ventilation were assessed clinically and by monitoring blood gases. No adverse effects were recorded. PaO2 was similar before and one hour after tiapride. There was no hypoventilation but, on the contrary, a slight decrease in PaCO2 (49,17 +/- 7,3 and 47,10 +/- 7,3 torr) and a significant increase in pH (7,358 +/- 0,023 and 7,378 +/- 0,028; p less than 0,05). Tiapride was also given to ten patients before fiberoptic bronchoscopy. Endoscopy was well tolerated in eight patients, in six there was no anxiety, and in all normal vigilance was retained. PMID- 6275546 TI - [Local and regional extension of breast carcinoma; report of one hundred and sixty-two cases. (author's transl)]. AB - Following mastectomy for cancer, one hundred and sixty-two biopsy samples were examined to determine the frequency of tumoral lesions other than those of the main tumor. If tumoral remnants close to the excision zone or less than five cm from the main tumor are eliminated, 44.4 per cent of cases had other remote tumors. In the present series, the frequency of other tumoral lesions was far from negligible for small tumors, and increased when the main tumor had intraductal components and when four or more axillary lymph nodes were invaded. These results, which confirm those of other authors, have to be considered for effective treatment of breast cancers, even those of a small size. PMID- 6275547 TI - [Severe digestive hemorrhage. Place and results of arterial embolization (author's transl)]. AB - Arterial embolization for hemostatic purposes in digestive hemorrhage was carried out in 11 cases of acute hemorrhage and in 2 cases of chronic hemorrhage. In all cases (11) in which selective embolization was possible, immediate hemostasis was obtained. The two hemorrhagic relapses observed later were due to carcinomatous lesions. There was only one serious complication in the form of ischemic stenosis after jejunal embolization. Embolization is the method of choice in all cases in which surgery is contra-indicated. Apart from this category of patients of which the prognosis remains bad in spite of hemostasis, there are other indications in which embolization may permit in spite of a reasonable risk, transformation of emergency surgery into cold surgery with lesser mortality. Finally, in a few cases hemostasis alone and medical treatment may prove to be sufficient and avoid any further surgical operation. PMID- 6275548 TI - [Partial progressive lipoatrophy. A new case-report (author's transl)]. AB - A new case of lipoatrophic diabetes with some peculiar features is reported. The disease was acquired, with onset at approximately thirty years of age, and partial, involving only the face and the upper limbs. The authors were able to observe the successive occurrence of the various symptoms: progressive lipoatrophy, hyperinsulinism, insulin-resistant diabetes, major lipid disturbances, and finally renal failure with a nephrotic syndrome and high blood pressure. Nosologic borders and physiopathology are discussed with a review of the literature. PMID- 6275549 TI - [Lingual necrosis with temporal arteritis (author's transl)]. PMID- 6275550 TI - [Pericarditis occurring in the course of meningococcal meningitis. Description of a case and review of literary data. (author's transl)]. AB - The authors report a new case of acute pericarditis in a 21 month-old girl, occurring on the fourth day of a meningococcal meningitis. The patient's recovery from the pericarditis was achieved thanks to a surgical drain in association with antibiotherapy, and despite a relapse after a puncture. On the basis of this case and the literary data reviewed, they endeavor to assess the frequency of this rare complication, to examine the most likely physiopathological hypothesis and to study the most effective treatment. PMID- 6275551 TI - [Glomerulonephritis in Henoch-Schoenlein purpura. Anatomo-clinical correlations and immunopathological study of twenty-five cases. (author's transl)]. AB - Twenty-five observations (children and adults) of glomerulo-nephritis secondary to Schoenlein-Henoch purpura were studied retrospectively. Histological studies done early in the course of the disease-less than one year after the first attack enabled us to made, in function of the evolution, clinical and pathological correlations using the method of Levy et al. The intensity of the clinical signs at the time of biopsy appeared proportional to the degree of cellular proliferation whether it was endo or extracapillary form; it is much more difficult to define when mesangial proliferation predominates or in the absence of proliferation. In addition to the usual immunopathological abnormalities, this study found that IgE plasma concentration was frequently elevated. Thus, IgE may play a role in the pathogenesis of this systemic disease. PMID- 6275552 TI - In vitro evaluation of anticancer drugs with the human tumor stem cell assay. PMID- 6275553 TI - [Methods of functional examination of peripheral nerves]. PMID- 6275554 TI - [Trophic disorders of neurologic origin. Perforating plantar disorders]. PMID- 6275555 TI - [Vulvar sclero-atrophic lichen. Bowen's disease and Paget's disease of the vulva]. PMID- 6275556 TI - Acute myelogenous leukemia after successful treatment of small cell carcinoma of the lung. PMID- 6275557 TI - [Current concepts on the aetio-pathogenesis of acute leukaemia (author's transl)]. PMID- 6275558 TI - [Biological and immunological markers in acute leukaemia (author's transl)]. PMID- 6275559 TI - Biochemical actions of anti-alcoholic agents. PMID- 6275560 TI - Assay of endogenous opiate receptor ligands in human CSF and plasma. PMID- 6275561 TI - Reversion from deficiency of galactose-1-phosphate uridylytransferase (GALT) in an SV40-transformed human fibroblast line. AB - Control SV40-transformed human fibroblasts can be readily adapted to growth on medium containing galactose as sole hexose source (galactose-MEH). However, most cells from a line of SV40-transformed skin fibroblasts from a patient with galactosemia (galactose-1-phosphate uridylyltransferase (GALT) deficiency) died in galactose-MEM. Surviving cells of this line either grew in completely sugar free media or had acquired significant amounts of GALT activity. Two presumptive revertant cell lines with GALT activity were characterized in detail. The expression of GALT in these two lines was stable in nonselective conditions. Each had different reaction maximum velocities with respect to uridine diphosphoglucose (UDPg) concentration as compared to residual activity in the parental cell strain or control cells. Both appeared to demonstrate heat inactivation profiles for GALT than differed from the parental cells or controls. UDPG concentration was found to significantly alter the thermostability of GALT. A competitive radioimmunoassay for GALT showed that these two lines had amounts of the GALT protein comparable to that of the parental cell strain or control cells. The electrophoretic mobility of GALT from the two presumptive revertants was found to differ from control cells. It was concluded that structural gene changes were probably responsible for the apparent reversion in these lines. PMID- 6275562 TI - Epstein-Barr (EB) virus antibodies in rheumatoid arthritis. PMID- 6275564 TI - Peripheral neuropathy: etiological, clinical laboratory and electrodiagnostic correlation. PMID- 6275563 TI - Naloxone reverses tissue effects of live Escherichia coli sepsis. AB - Hypoxia of the superficial gastric epithelium induced by a systemic infusion of live Escherichia coli organisms was mimicked by a local intra-arterial infusion of the naturally occurring opiate beta-endorphin. Naloxone, a specific opiate antagonist, reversed the gastric epithelial hypoxia induced by sepsis and also prevented the development of systemic acidosis. The mean blood pH of septic dogs had declined during the experiment from 7.42 +/- 0.06 to 6.88 +/- 0.17, whereas corresponding values for the naloxone-treated group were 7.38 +/- 0.06 and 7.32 +/- 0.08. These experiments, which support the concept of beta-endorphin involvement in the pathogenesis of septic shock, indicate a direct tissue response to circulatory beta-endorphin and highlight a further beneficial effect of naloxone in the management of sepsis. PMID- 6275565 TI - The physical chemistry of the gelation of sickle hemoglobin. PMID- 6275566 TI - Prenatal diagnosis of sickle cell anemia and beta-thalassemia by amniocentesis. AB - Prenatal diagnosis by amniocentesis alone is possible for about 90% of pregnancies at risk for sickle cell anemia and about 70-75% of pregnancies at risk for beta-thalassemia. It should be stressed that to obtain these percentages, a previous homozygous normal or affected child or, alternatively, the couple's parents are needed to confirm the linkage of variant genes to respective DNA markers (polymorphic restriction sites). Families at risk should be studied prior to or early in pregnancy to determine whether these methods will be applicable to their specific case. If appropriate markers are identified and their linkage to the genes being studied is verified, then prenatal diagnosis by amniocentesis can be done at 16-18 weeks; and for these couples, fetoscopy, with its increased risk of fetal loss, can be avoided. The change of errors in diagnosis due to crossing-over between the mutant gene and the linked marker is small. The probability of such a recombination between the beta-globin gene and the polymorphic G gamma Hind III or the 3' Hpa I site is 1/3,000 and 1/14,000 per generation, respectively (13). A more serious problem is non-paternity because of the errors caused in linkage analysis. Improved methods to enable direct detection of the beta S and/or beta thal mutations would remove both of these potential sources of error. PMID- 6275567 TI - Some biochemical activities of 10,10-difluoro-13-dehydroprostacyclin, a chemically stable analog of prostacyclin, in human blood platelets. PMID- 6275568 TI - A peptide derived from fibrin(ogen) inhibits angiotensin converting enzyme and potentiates the effects of bradykinin. PMID- 6275569 TI - [Recombinant DNA and genetic manipulation, reality and fantasy (author's transl)]. AB - Genetic manipulation deliberately tries to influence the progeny of a living organism to achieve a human purpose. Well-known instances are artificial insemination and embryo transfer. The species barrier is not broken in this procedure and a surrogate mother is required in every case. In frogs, the nuclei og a number of fertilized ova may be replaced by the nuclei of vegetative cells of a single frog (a) and thus produce a clone of identical frogs (a). This is impossible in mammals. Parts of plants and even cultures of vegetative plant cells may develop into complete plants. This fact is gratefully used for the rapid propagation of useful hybrids. The recombinant DNA procedure is a method by which particles of any DNA can be multiplied in a culture of bacteria. This DNA can consequently be cloned. The basic principles of the procedure are discussed. When the recombinant DNA technique is used, the species barrier is passed. It can only be used, however, to influence the heritable characteristics of unicellular organisms or vegetative cells in cultures. This may possibly include plants. The risks run by the investigator using this technique are much smaller than those to which he is exposed in the case of pathogenic bacteria. When non-pathogenic bacteria showing a high degree of biological containment are used, the hazard of spreading in the environment is small. This problem undoubtedly merits further attention in other organisms. Mammals cannot be 'clonized' or freaks produced using recombinant DNA procedures. PMID- 6275570 TI - [Information about the social-psychiatric service]. PMID- 6275571 TI - Gentamicin nephrotoxicity. I. Acute biochemical correlates in rats. PMID- 6275572 TI - Gentamicin nephrotoxicity. II. Plasma membrane changes. PMID- 6275573 TI - Aryl hydrocarbon hydroxylase (AHH) induction by polybrominated biphenyls (PBBs): enchancement by photolysis. AB - Irradiation of the commercial polybrominated biphenyl (PBB( mixture, fireMaster BP-6, in cyclohexane solution at 300 nm for 930 min resulted in a marked diminution of the major components of the mixture. Administration of the photolyzed PBB mixture of fireMaster BP-6 to immature male Wistar rats caused both dose-related decreases in thymus weight and increase in hepatic microsomal benzo[a]pyrene hydroxylase (AHH), 4-dimethylaminoantipyrine N-demethylase and NADPH-cytochrome c reductase activities and cytochrome P-450 content. The dose effecting half-maximal AHH induction for the photolyzed PBBs (9 mg . KG-1) was approximately 6 times lower than that of fireMaster BP-6 (50 mg. kg-1). Furthermore, the concentration of photolyzed PBBs (2 micrometers) required to displace 50% of the specifically-bound [3H] TCDD from its high-affinity cytosolic Ah receptor was approximately 150 times lower than that required for fireMaster BP-6 (300 micrometers), as measured by sucrose density gradient centrifugation analysis. The results suggest that the photolysis of the commercial PBB mixture yields products which possess increased biologic activity. PMID- 6275574 TI - Acute type B viral hepatitis due to accidental transfusion of blood containing HBs Ag in a patient with liver cirrhosis. AB - Patients with liver cirrhosis have the opportunity of receiving blood transfusions rather frequently. Accidental transfusion of blood containing HBs Ag occurred in such a case. Recently, we encountered a case of hepatocellular carcinoma based on posthepatitic cirrhosis in which an emergency blood transfusion because of massive hematemesis from a gastric ulcer was followed by acute B type viral hepatitis. The remainder of the transfused blood was preserved an we tested it serologically when the symptoms of acute viral hepatitis were manifested. We are able to detect hepatitis B surface antigen in the serum. In this patient, the preceding liver cirrhosis did not influence on the clinical course of acute B type viral hepatitis and conversely, the acute B type viral hepatitis did not have any influence on the subsequent clinical course of the liver cirrhosis. PMID- 6275575 TI - Derivatization of estrogen conjugates for analysis by capillary gas chromatography. AB - By using 20 meter wall-coated open tubular glass capillary columns of high stability, analysis of methyl ester, methyloxime trimethylsilyl ether derivatives of estrogen glucuronides had been achieved. Relative retention times of five glucuronide conjugates on OV-1 stationary phase are reported. Estrogen sulfates conjugated at the 3-position were shown to be quantitatively hydrolyzed and derivatized in a single trimethylsilylation step, and this method of direct derivatization was compared to two solvolysis methods. These analytical methods could be further developed to allow rapid and quantitative analysis of estrogens in biological fluids, and may prove particularly useful for analysis of labile compounds. PMID- 6275576 TI - A short efficient synthesis of 16-oxygenated estratriene 3-sulfates. AB - A novel synthesis of sodium 17-oxo-16 alpha-hydroxy-1,3,5(10)-estratrien-3-yl sulfate (4), sodium 16 alpha, 16 beta-dihydroxy-1,3,5(10)-estratrien-3-yl sulfate (5) and sodium 16-oxo-17 beta-hydroxy-1,3,5(10)-estratrien-3-yl sulfate (6) is described. 16 alpha-Bromo-3-hydroxy-1,3,5(10)-estratrien-17-one (1) was efficiently synthesized in one step with 70-97% yield by bromination of 3-hydroxy 1,3,5(10)-estratrien-17-one with cupric bromide. 3,16 alpha-Dihydroxy-1,3,5(10) estratrien-17-one (3) was quantitatively obtained by controlled stereospecific hydrolysis of the bromoketone 1 with sodium hydroxide in aqueous pyridine. The bromoketone 1 was converted to the 16 alpha-hydroxy-17-ketone 3-sulfate 4 by sulfation with chlorosulfonic acid in pyridine and a subsequent controlled hydrolysis in a high yield without formation of the other ketols. Treatment of the sulfate 4 with sodium borohydride have the triol sulfate 5. The sulfate 4 was also rearranged to the 17 beta-hydroxy-16-ketone 6 with sodium hydroxide in water in a quantitative yield. PMID- 6275577 TI - Zika virus, a cause of fever in Central Java, Indonesia. AB - In 1977 and 1978 selected in-patients at the Tegalyoso Hospital, Klaten, Indonesia who had recent onsets of acute fever were serologically studied for evidence for alphavirus and flavivirus infections. A brief clinical history was taken and a check list of signs and symptoms was completed on admission. Acute and convalescent phase sera from 30 patients who showed evidence that a flavivirus had caused their illnesses were tested for neutralizing antibodies to several flaviviruses which occur in South-east Asia. Paired sera from seven patients demonstrated a fourfold rise in antibody titre from acute to convalescent phase. The most common clinical manifestations observed in this series of patients included high fever, malaise, stomach ache, dizziness and anorexia. None of the seven patients had headache or rash despite the fact that headache and rash had been associated with two of the three previously studied. The onsets of illness clustered toward the end of the rainy season when populations of Aedes aegypti, a probable vector in Malaysia, were most abundant. PMID- 6275578 TI - An epidemic of rotavirus diarrhoea in Manipur, India. AB - Recurrent epidemics of acute diarrhoeal disease in young children have occurred during the winter months in Manipur (India) for many years. During November 1979, children admitted to hospital for acute diarrhoea were investigated and, in addition to pathogenic Escherichia coli and Shigella rotovirus was detected by ELISA in a number of stool samples, especially of those less than one year old. The importance of this finding and the need for further study are discussed. PMID- 6275579 TI - Allopurinol in the treatment of visceral leishmaniasis. AB - Allopurinol was used in the treatment of 10 patients with kala-azar. Of six patients who had previously failed to respond satisfactorily to Pentostam, "cures" were achieved in four. However, it was necessary to add Pentostam to the allopurinol in one, and another relapsed after apparent "cure" but again responded to allopurinol. The response of four patients who had had no previous treatment for kala-azar was less satisfactory. PMID- 6275580 TI - The spectrum of liver disease in the United Arab Emirates. AB - Of 209 patients with liver disease attending the Al Qassimi Hospital, Sharjah, 172 were suffering from acute hepatitis. The proportion harbouring the hepatitis B surface antigen was unknown. Of the remainder, 10 were suffering from chronic active hepatitis, five from primary hepatoma and six from cirrhosis. Infection with hepatitis virus B was regarded as of aetiological significance in three cases of chronic active hepatitis, four of primary hepatoma and two of cirrhosis. PMID- 6275581 TI - Use of enzyme linked immunosorbent assay in intestinal and extra-intestinal amoebiasis (amoebic liver abscess). AB - The sera of patients with amoebic liver abscess, intestinal amoebiasis and non specific hepatomegaly were examined for the presence of antibodies to Entamoeba histolytica antigen by enzyme linked immunosorbent assay (ELISA). All the amoebic liver abscess sera were strongly positive. The test was also positive in all the proved cases of intestinal amoebiasis but the ELISA values were relatively slightly lower. Nine of 10 cases of non-specific hepatomegaly with stools negative for E. histolytica gave positive results with ELISA. The IHA titre in all but one of the non-specific hepatomegaly cases was within normal value. PMID- 6275582 TI - [Cytogenetic action of the discharges from superphosphate manufacture on animals]. AB - Susceptibility of animal chromosome apparatus to a prolonged action of industrial superphosphate pollutants was investigated by registration of structural mutations in the bone marrow cells of the yellow gopher at the anaphase and metaphase of mitosis. A significant rise in chromosomal aberration frequency is shown for rodents caught in the technogenic province surrounding the works. PMID- 6275583 TI - [Genome instability and neoplasia]. AB - Necessity of oncogene for the neoplastic cell transformation is substantiated. The oncogene may be introduced into the cell by a virus; be present in the cell from its formation but be repressed by products of regulatory genes; be arisen from separated DNA segments. The latter possibility is discussed in view of data on the presence of mobile elements and Sarc sequences in the cell genome. PMID- 6275584 TI - [Effect of cortisol, adrenaline and ACTH on the absorption and intracellular distribution of protein-labelled lipoproteins by rat liver and adrenal tissues]. AB - One hour incubation of slices prepared from rat liver or adrenal glands with 125I labeled lipoproteins of different classes resulted in label binding and its penetration into the tissue cells. The greatest amounts of labeling were found in fractions of mitochondria, lysosomes, microsomes and the final supernatant. In relation to the weight unit, slices from adrenal glands consumed several times as much label as did liver slices. Epinephrine increased the amount of the label bound to the low density lipoproteins by several times in the fractions of large cell membranes, and decreased the 125I level in mitochondria. Cortizole increased the incorporation of the protein labeled lipoproteins very low density into the liver mitochondria. ACTH did not influence the incorporation or cellular distribution of the label in adrenal glands. PMID- 6275585 TI - Levels of polyamines and nucleic acids in human breast carcinoma. AB - Polyamine and nucleic acid levels were determined in normal human breasts, in primary infiltrating ductal carcinoma breasts, in the uninvolved tissues of the same carcinoma-bearing breasts and in the urine of the same patients. The results showed that the production of polyamines in the carcinoma tissue is significantly higher than in normal tissue; the presence of a carcinoma in the breast does not modify the levels of polyamines and nucleic acids in the surrounding uninvolved tissue; spermidine and spermine are correlated both with DNA and RNA in the carcinoma; the urinary polyamine levels in the carcinoma-bearing patients were not significantly different from that of controls. There appeared to be a close relationship between polyamine biosynthesis and tumoral mammary cell development and a very fine control between the biosynthesis of both spermidine and spermine and nucleic acids. In addition, the localized solid breast carcinoma, with non proven metastases, did not provoke an increase in polyamine excretion. PMID- 6275587 TI - Malignant mixed Mullerian tumors of the uterus. A clinical and pathologic study of 34 cases. PMID- 6275586 TI - Unexpected breast cancer detection after outpatient biopsy for benign lesion. AB - At the Istituto Nazionale Tumori of Milan, in its Outpatient Department, 1302 breast surgical biopsies were carried out in 1978. Fibroadenomas occurred in 44.2% and benign dysplasias in 39.3%. Sixty-two cancers (4.8%) and 23 atypical hyperplasias (1.8%) were unexpected findings, with an increasing incidence in women over 50. Seventy-nine duct resections were performed for significant nipple discharge and revealed 5 unexpected carcinomas and 28 papillomas or papillomatosis. In our opinion and experience, this procedure of performing breast biopsy in the presence of a solid lump in women over 30 without hospitalization improves early detection of cancer, lowers costs, cuts down admission waiting lists and increases the number of women who better accept a surgical diagnosis and treatment. PMID- 6275588 TI - High dose BCNU followed by autologous bone marrow infusion in glioblastoma multiforme. PMID- 6275589 TI - [Activity of Krebs cycle dehydrogenase and tissue respiration enzymes in cerebral hemispheres of rats under stress]. AB - It is established that emotional-painful stress and myocardium necrosis evoked in animals after the emotional-painful stress is accompanied by a disturbance in the creative metabolism and partial inhibition of cytochrome oxidase in cerebral hemispheres. A conclusion is made that the found changes may be one of the pathogenetic factors in formation of the neurosis-like states under stress and diseases originated from the effect of the emotional-psychic overstrain. PMID- 6275590 TI - [Specific and metabolic effect of oxythiamine]. AB - Increasing doses of oxythiamine were studied as exerting the effect on transketolase inactivation in rat tissues. A conclusion is made that in the process of synthesis de novo there is a transient form of the enzyme accessible for interaction with oxythiamine pyrophosphate. Injection of oxythiamine in the increasing doses are accompanied by a decrease in the glycogen amount, increase in the intracellular level most of the studied intermediates of glycolysis and pentose cycle as well as cAMP. The probable biochemical mechanism of the oxythiamine action is connected with the activation of processes dependent on cAMP. PMID- 6275591 TI - Ultrastructural and Energy dispersive analysis of inorganic inclusions in a muscle biopsy. AB - A muscle biopsy that, by light microscopy, exhibited mild atrophy consistent with chronic denervating disease was submitted for ultrastructural analysis. Inorganic structures within the tissue were defined by energy dispersive analysis as aluminosilicates, magnesium silicates, and iron deposits. These structures were localized in the interstitial (intercollagenous) area and in high concentrations within degenerated muscle bundles. An alteration of the blood/muscle barrier has obviously occurred; however, the extent to which the end result commonly occurs in humans is unknown. Clarification of the question raised by this observation will have to be derived from animal studies, and clinicians must have an awareness of the need for electron microscopy/energy-dispersive x-ray analysis in muscle biopsies from patients with similar therapeutic backgrounds. PMID- 6275593 TI - Inclusions in Bone Marrow Cells. PMID- 6275592 TI - Ultrastructural and Immunological methods in diagnostic pathology in a community hospital. AB - Five unusual tumors and one case of glomerulonephritis were examined by light microscopy, immunochemical methods, and electron microscopy in order to arrive at a correct diagnosis. We propose that these techniques can be used in community hospitals for selected cases in order to classify certain types of tumors, identify the primary site of some metastatic tumors, and help in the precise classification of some autoimmune diseases. PMID- 6275594 TI - [Verner-Morrison syndrome]. PMID- 6275595 TI - Signet-ring cell carcinoma involving the urinary bladder. Report of a case and review of 21 cases. AB - A case of signet-ring cell carcinoma involving the urinary bladder, which is a rare variant of adenocarcinoma, is presented. 21 cases have so far been reported in the occidental and Japanese journals. In 13 patients (62%) the carcinomas were of urachal origin and 8 (38%) of bladder origin. The prognosis of signet-ring cell carcinoma originating in bladder is poorer compared to that in urachus; 2 year survival of 40% for the bladder cancer group and 70% for the urachus. A radical cystectomy with excision of adjacent tissues, which might improve the prognosis in this fatal malignancy, should be considered. PMID- 6275596 TI - [Transmission of Marek's disease to wild feathered game]. AB - The following wild feathered game were tested from 6 to 12 months of age: common pheasant (Phasianus colchicus), common gray partridge (Perdix perdix), European quail (Coturnix coturnix), and Greek partridge (Alectoris graeca). Only pheasant was found to be susceptible to infection with a virulent virus of Marek's disease. In this bird the first clinical symptoms of the disease (nervous changes) were observed in 28 days from infection. Immunoprecipitation antibodies to the Marek's disease virus occurred in the serum on the 75th day from infection, and patho-morphological changes, typical of acute Marek's disease, were observed on the 75th to 85th day from infection. The remaining species of wild feathered game appeared not to be susceptible to infection with the Marek's disease virus. In these birds, like in pheasants, immunoprecipitation antigens of the Marek's disease virus were not found to be present in the epithelial cells of feather follicles. In the infected Greek partridge, common gray partridge and quail, serum was not found to contain any antibodies. Laying quail had no antibodies in the yolk of the eggs they laid. The results indicate a different oncogenesis of Marek's disease in the domestic fowl and pheasant. It is concluded that wild feathered game play no important role in the epizootology of Marek's disease. PMID- 6275597 TI - [Electron microscopy of parvoviruses and rotaviruses in canine enteritis]. PMID- 6275598 TI - Parapoxvirus infection of the red squirrel (Sciurus vulgaris). PMID- 6275600 TI - Virus-like particles associated with disease in guinea fowl. PMID- 6275599 TI - Field and laboratory studies of equine influenza viruses isolated in 1979. AB - Experimental ponies developed signs of disease four days after the intranasal instillation of A/England 1/79 equine influenza virus and virus was recovered from the nasopharynx from the second to the ninth day. No significant antigenic difference was found between the virus and the prototype A/Miami 1/63 virus, using post infection ferret and chicken sera and post vaccination pony sera. No antigenic differences were found between four viruses isolated between January and July 1979, although some differences were found in their ability to detect haemagglutination inhibiting antibody in convalescent horse sera. PMID- 6275601 TI - Diazoxide treatment of canine insulinoma. PMID- 6275602 TI - Propagation of bovine rotavirus by dogs. PMID- 6275603 TI - [Part played by bovine adenoviruses in calf diseases]. AB - Parallel investigations were carried out on twice obtained blood serum samples (at 15-20 day intervals) from a total of 176 affected calves, exhibiting a respiratory syndrome on 20 cattle breeding farms, with the use of the immunodiffusion test with antigens obtained from prototypic strains of B-10 and Misk/67 bovine adenoviruses (1st serotype for 1st subgroup and 8h serotype for 2nd subgroup). In 16 out of 20 recorded enzootics adenoviruses participated in a varying degree in the etiology of the disease. In 23.28 per cent of the diseases calves there were newly appearing antibodies against the group-specific adenovirus antigens. In 14.20 per cent of the animals the antibodies were against 1st antigen subgroup, and in 9.03 per cent of them the antibodies were against the antigen of the 2nd subgroup. In 6 of the enzootics the relative share of adenoviruses was shown to be considerable (up to 40 per cent), and in 11 of them their participation was only partial. PMID- 6275604 TI - [Combined method (suspension-monolayer) of preparing an inactivated foot-and mouth disease vaccine]. AB - Semiproductional and productional experiments were carried out to cultivate BHK21C13 cells in 30- and 250-litre fermentors, to produce F.M.D. virus in suspension and in roller, and to produce a F.M.D. vaccine in 500-litre automatic mixing containers, employing a combined method 'suspension-monolayer'. The F.M.D. virus that was replicated in suspension had a 10(-6,0-6,5) titer, and the virus that was obtained in monolayer - a 10(-7,0-8,0) titer. It was demonstrated that the F.M.D. vaccines that were produced after the combined method were sterile, innocuous, and immunogenic. PMID- 6275605 TI - [Rotaviruses as a cause of an epidemic of acute gastroenteritis]. PMID- 6275606 TI - [Difficulties and errors in the cytological diagnosis of breast cancer]. AB - The paper deals with retrospective evaluation of cytograms of tumor punctates from 142 patients with breast cancer. It was found that adequate diagnosis is hard to make in cases of lobular and scirrhous carcinoma due to indistinct atypia of cells and nuclei. Errors in cytological diagnosis may arise both from absence of cell structure in tapping material and misinterpretation of morphological features of malignant cells in scirrhous carcinoma and highly-differentiated lobular cancer. PMID- 6275608 TI - [Case of the diffuse form of pulmonary adenomatosis with a lupus syndrome]. PMID- 6275607 TI - [Diagnostic value of determining chorionic gonadotropin and trophoblastic beta 1 glycoprotein in trophoblastic disease]. AB - The diagnostic value of determinations of the levels of chorionic gonadotropin (HCG) and trophoblastic beta 1-glycoprotein (TBG) was studied in 160 cases of trophoblastic disease. Trophoblastic disease was diagnosed in 86% of cases, when HCG level was determined immunologically and in 100 % -- by radioimmunological means and in 86 and 33% when TBG level was established by immunoradioautographical and immunodiffusion methods, respectively. The results of HCG and TBG assays by immunological and immunoradioautographical methods matched in 96% of cases. The said methods are highly specific and may find a wide application in diagnosing trophoblastic disease. The radioimmunological method of HCG assay is the most sensitive. However, in view of possible HCG synthesis in cases of other tumor, its application should be preferred in assessing the efficacy of therapy rather than for primary diagnosis of trophoblastic disease. PMID- 6275609 TI - [Characteristics of collagen protein biosynthesis in fibroblast cultures from the skin of patients with systemic scleroderma]. PMID- 6275610 TI - [Effect of rations containing fluorine ion on biochemical and hematological indices of body status]. AB - A study was made of the fluorine-ion content in the vegetables and grain grown on experimental plots of land fertilized for 4-6 years with fluorine-containing fertilizers: active slurry potassium tetrafluoroborate and sodium fluorosilicate. It was found that the fluorine-ion content in individual samples of the vegetables and grain was increased 2-32-fold as compared to that in the control samples. As for the diet, the total content of fluorine-ion was increased 6.1 12.3-fold. Experiments were made on rabbits and rats whose diets contained the vegetables and grain from the experimental plots of land. The animals of the experimental groups manifested a lowering of the prothrombin index by the end of the 6th month of the experimental studies. There was a statistically significant fall in the cholinesterase activity and an increase in the activity of glutaminopyruvic aminotransferase (by 35-42 units). The activity of total alkaline phosphatase progressively increased at the expense of the elevated activity of liver and intestinal isozymes. PMID- 6275611 TI - [Biliary tract carcinomas. II. Biliary tract cancer]. AB - The authors discuss the biliary tract carcinoma in the second paper on biliary tract carcinomas. It ranks second as regards frequency, being responsible for 4 per cent of the biliary operations. The analysis is based on 48 observations during the period 1952-1979 at the Chair of Abdominal Surgery - 31 males and 17 females (2:1). Morbidity rate is the highest among the patients aged from 40 to 60. The patients are admitted for operation up to the 1-2 months since the beginning of their complaints, mostly due to the lasting jaundice, present in all the patients (100%). Regardless of the application of the present methods, biliary ducts carcinoma remains a serious diagnostic and surgical problem. Those methods, however, are the only possible ones, for an earlier detection of the tumors, hence, for the improvement of the unsatisfactory, so far, operability, lethality and survival. PMID- 6275612 TI - [Clinical results with a complex conservative treatment method in infiltrative ophthalmopathy (preliminary report)]. AB - The present methods for the treatment of infiltrative ophthalmopathy are still unsatisfactory, some of them associated with considerable and lasting side effects. Clinical results are present in the communication, from the application of an original therapeutic complex, consisting in the parallel administration of ACTH, intravenous infusions, moderate doses cortison, diuretics and sanation via antibiotics and eventual surgical intervention of the existing focal infectious agents. The complex is applied in the course of 4 weeks. Regression of the clinical symptomatics is observed and assessed by a particular method, it reaches an average of about 50 per cent of the initial manifestations. In 45 per cent of the patients the result was assessed as very good and excellent. The subjective complaints, congestion, chemosis and edema are best and most rapidly influenced, whereas exophthalmos and the eventual ophthalmoplegic phenomena showed no essential changes. The correlations between the results obtained, accompanying the disease (Basedow disease, focal infections) and the severity of ophthalmopathy were discussed. The method was admitted to be perspective, harmless and could ensure good and lasting therapeutic results of properly and persistently applied. PMID- 6275613 TI - [Epidemiological role of Blattella germanica (L.) in amebiasis]. PMID- 6275614 TI - [Development and results of adrenal autotransplantation (author's transl)]. AB - 56 cases of adrenalectomy and autotransplantation of human adrenal tissue have been reported in the literature since 1955. The most common indication was bilateral adrenal hyperplasia. A reduction in postoperative corticosteroid substitution and a return to almost physiological function on discontinuation of long-term substitution can be achieved. Recurrent disease is easily cured by partial excision of the transplanted tissue, situated usually in small muscle pockets, under local anaesthesia. The loss of adaptation to circadian rhythm and stress are adverse effects. Further development of this technique, which is still relatively rarely implemented, may make it more universally applicable for a wider indication of bilateral adrenalectomy, without loss of adrenal function. PMID- 6275615 TI - [Preoperative diagnosis of a Meckel's diverticulum (author's transl)]. AB - The most common complications of a Meckel's diverticulum is bleeding. With the introduction of abdominal scintigraphy we have a non-invasive examination method at our disposal which enables this condition to be diagnosed with accuracy preoperatively. A relevant case is described. PMID- 6275616 TI - Activation of benzo(a)pyrene and 2-acetamidofluorene to mutagens by microsomal preparations from different animal species: role of cytochrome P-450 and P-448. AB - 1. The metabolic activation of benzo(a)pyrene and 2-acetamidofluorene to mutagens was studied with liver microsomal preparations from rat, guinea-pig, hamster and mouse, untreated or pretreated with phenobarbitone or 3-methylcholanthrene. 2. Liver microsomal preparations from all animal species activated benzo(a)pyrene, that from mouse being the most efficient. Similarly, microsomal preparations from guinea-pig, hamster and mouse could activate 2-acetamidofluorene, but that from rat exhibited very weak activity. 3. Activation of benzo(a)pyrene into mutagenic intermediates by liver microsomal preparations was increased for all animals except mouse by pretreatment with 3-methylcholanthrene. In contrast, pretreatment with phenobarbitone decreased the activation by microsomal preparations from all species. 4. Activation of 2-acetamidofluorene by liver microsomal preparations from rat and guinea-pig, but not mouse and hamster, was increased by pretreatment of the animals with phenobarbitone. Pretreatment with 3-methylcholanthrene decreased the activation of this carcinogen by microsomal preparations from all species. 5. The metabolic activation of benzo(a)pyrene is catalysed by cytochrome P-448 but not cytochrome P-450. 6. The activation of 2-acetamidofluorene to mutagens may involve, in addition to the mixed-function oxidases, other microsomal enzyme systems. PMID- 6275617 TI - Chemotherapy of lung cancer. AB - The potential for substantial improvement in the outcome of patients with carcinoma of the lung seem most likely to develop in the field of chemotherapy. In the past decade, striking advances in the management of small cell carcinoma have yielded response rates and longer survival. While the greatest improvement can be predicted for patients whose disease is limited in extent, combination chemotherapy and combined modality therapy generally are effective in causing tumor regression for the majority of patients. About 20 percent of patients with disease limited to the thorax and lymph nodes will survive two years. In non small cell tumors, response rates are improved with intensive drug combinations, although the majority of cases are unresponsive to present regimens. Careful staging and evaluation of patients indicates that patients with good performance status and limited extent of disease appear to obtain the most benefit from intensive treatment. The considerable morbidity of some treatments often influences the choice for or against chemotherapy in patients with non-small cell carcinomas. For the future, problems of particular interest will be in investigation of factors-biologic, pharmacokinetic, immunologic-that are related to the failure to cure small cell carcinoma, the most therapeutically responsive pulmonary tumor. Additionally, in the non-small cell tumors, more effective therapies as well as clarification of the basis for relative resistance to cytotoxic agents are areas for intensive investigation. PMID- 6275618 TI - Effect of lithium chloride on the circadian rhythm in the flight activity of the microchiropteran bat, Taphozous melanopogon. AB - We have performed experiments on the influence of Li+ offered through drinking water i) on the period length (tau) of the rhythm in the flight of bats under LL and ii) on phase angles during entrainment under LD cycles. Bats subject to entrainment exhibited no qualitative and quantitative change in psi values after Li+ treatment. In contrast the ingestion of Li ions by bats under free-running conditions shortens the tau of the bat rhythm by an amount that is a function of period length prior to ingestion. Tau is, thus homeostatically conserved in the face of Li administration in bats. PMID- 6275619 TI - Altered heme environments in opossum and rabbit methemoglobins. AB - The effect of pH and inositol hexaphosphate (IHP) on the symmetry of the heme environments in opossum (Didelphius marsupialis) and new Zealand White rabbit hemoglobins has been studied using electron spin resonance (ESR). Each methemoglobin is found to contain two different heme environments as detected by the rhombicity observed in the ESR spectrum. In both cases the rhombic nature of the ESR spectrum is influenced by pH and IHP binding, although in the case of rabbit methemoglobin the high spin ESR signal disappears above pH 8.0. In both hemoglobins, amino acid alterations in the alpha-chains are known to affect the properties of the ferrous derivatives. It is concluded that these alterations also provide the basis for the ESR spectral properties observed with the methemoglobins. PMID- 6275620 TI - A family of sequences with regional homology to bovine 1.715 satellite DNA. AB - The organization of sequences homologous to the bovine 1.715 satellite DNA was studied in the bovine genome. A novel family of sequences, containing different numbers of copies of the 1.4 kb satellite period alternating with 1.2 kb sequence not related to this satellite, was found. Possible mechanism of the generation of these sequences is proposed. PMID- 6275621 TI - Different protein-lipid interaction in human red blood cell membrane of Rh positive and Rh negative blood compared with Rhnull. AB - 1-anilino-naphthalene-8-sulfonate (ANS) fluorescence measurements have revealed that red blood cell membrane of the Rhnull type undergoes a transition at about 16 degrees C. In contrast, viscosity measurements of the extracted membrane lipids showed the usually observed transition at about 18 degrees C. Lower values of titratable sulfhydryl (SH) groups were observed in Rhnull membrane using 5,5' dithiobis-(2-nitro-benzoic-acid) (Nbs2). In contrast, disulfide bonds in Rhnull membrane were estimated to be about 3 times the value of the controls. Spin labeling experiments using 2-(3-carboxypropyl)-4, 4 dimethyl-2-tridecyl 3 oxazolidinyloxyl were carried out with phospholipase A2 modified membranes. The mobile part of the spectra was significantly increased on the Rhnull membrane. In the presence of D-glucose, infrared spectrometry showed a larger reduction of the intensity of the POO-band in Rhnull membrane. In contrast to controls, binding of the reagent diethylpyrocarbonate resulted in no significant changes of the Rhnull membrane as determined by electron spin resonance (ESR) measurements. D-glucose transport activity was found to be at the upper level of a group of Rh positive and Rh negative persons. It is suggested that the intensity of the polar protein lipid interaction is reduced in Rhnull membrane. PMID- 6275622 TI - [Experimental gene transfer in mammalian somatic cells]. PMID- 6275623 TI - [Migrating and insertion sequences of DNA]. PMID- 6275624 TI - [Intercellular contacts of epithelial layers: the physiological significance and their potential regulation]. PMID- 6275626 TI - Detection of different types of receptors for deoxyribonucleic acid in competent Bacillus subtilis. AB - Competent cells of Bacillus subtilis are able to bind but not to take up DNA in magnesium-free medium. Wall-membrane complexes, but not cell walls, bind homologous DNA. Binding of DNA to membrane vesicles suggests that DNA-receptor sites are located at the membrane level. On the basis of competition experiments with homologous and heterologous DNAs, in cells and in membrane vesicles, different types of DNA receptors are detected. PMID- 6275625 TI - [Postoperative treatment of malignant glial tumors of the brain with CCNU]. PMID- 6275627 TI - [Benign and semimalignant soft tissue tumours (author's transl)]. AB - 1033 cases of benign and semimalignant tumours of the soft somatic tissue are examined. Tumours of the fatty tissue and those of blood vessels do not cause any difficulties in diagnostics and therapy. Concerning semimalignant tumours there is a high risk of recidivation but no metastasising. Likewise there is a high risk of recidivation in tumours of the fibrillar and embryomorphous tissue. Patients suffering from these tumours are included in the aftercare programme corresponding to the postoperative treatment of patients with malignant tumours. PMID- 6275628 TI - [Cylindroma of the trachea]. PMID- 6275629 TI - [The virus inactivating efficacy of peracids and peracidous disinfectants (author's transl)]. AB - It has always been an aim to develop chemicals able to kill a broad spectrum of microorganisms. Research in this field led to peracids inactivating vegetative bacterial forms, spores, fungi and viruses. Since liquid peracids are very unstable, powders were developed which dissolved in water, formed peracids. In this paper the inactivating efficacy of peracids derived from powders as well as of pure peracids is presented. It is shown that both inactivate the following viruses: poliomyelitisvirus type 1, coxsackievirus B3, adenovirus type 5 and SV40. The most resistant viruses were the picornaviruses, especially coxsackievirus B3. But in all cases, inactivation is in accordance with the guidelines of the Deutsche Vereinigung zur Bekampfung der Viruskrankheiten (DVV) (15). Furthermore, it is shown that pure peracids are more reactive than peracids formed from powders. But with all peracids there remains residual infectivity, which may be due to inhomogenic virus populations. PMID- 6275630 TI - An outbreak of Clostridium perfringens Hobbs type 21 food poisoning. PMID- 6275631 TI - Investigations on viral pollution in the Romanian section of the Danube river during 1972-1977 period. PMID- 6275632 TI - [Microbial resistance to formaldehyde. II. Dependence of the microbicidal effect upon the concentration of and the period of exposure to formaldehyde (author's transl)]. PMID- 6275633 TI - [Relative effectiveness of disinfectants against rotaviruses (author's transl)]. PMID- 6275634 TI - Dispersal and fate of coliphages in the River Saar. AB - The dispersal of coliphages was analysed quantitatively in longitudinal profiles of the partially canalized River Saar. The numbers of coliphages ranged between 0 and 2380 PFU (plaque-forming units)/ml in the water and between 0 and 2550 PFU/g in the sediments, depending on the degree of fecal pollution. In canalized parts the phages were found mainly in the sediments, whereas in non-canalized parts they prevailed in the water phase (Fig. 3). Determinations performed at municipal sewage water outfalls revealed that in the canalized part most of the phages precipitated to the sediment on a water course of only about 250 m (Fig. 5). In the non-canalized part of the River Saar, sediments were not contaminated to this extent, since phages rather remained in the water, due to high current velocities (Fig. 5). According to these results coliphages are, just like some fecal bacteria, closely associated with particles. They are rapidly inactivated in the sediment and water (inactivation rates: 0.12 to 0.66 d-1). PMID- 6275635 TI - [Carcino-embryonic antigen in cases of cervical carcinoma (author's transl)]. AB - Carcino-embryonic antigen (CEA) was determined by means of solid-phase radio immuno-assay through one year in 83 patients with Groups I through IV cervical carcinoma. The intention was to test CEA diagnosis for its value as tumour marker in therapeutic monitoring and detection of recurrences. CEA values prior to treatment were above normal, that is in excess of 2.5 ng/ml, in 42.8 per cent of the cervical carcinoma patients. CEA values declined during therapeutic radium and cobalt 60 irradiation in 81 per cent of those patients in whom original values had been increased to more than 2.5 ng/ml. No substantive change was recordable from those patients in whom original values had been below 2.5 ng/ml. Recurrence diagnosis gave good agreement between CEA levels and clinical findings in 58 per cent of the cases, falsely negative data in 37 per cent and falsely positive values in four per cent. The development of recurrence was repeatedly sensed by means of CEA determination up to six months before clinical manifestation in 72 patients who had been under long-time monitoring. The usefulness of CEA determination in therapeutic monitoring and after-care of patients with cervical carcinoma was found to be limited to the effect that only increased values were of clinical relevance, whereas no conclusions whatever could be drawn from low values. PMID- 6275636 TI - Prevalence of bluetongue virus precipitating antibodies in sheep and goats of Punjab, India. PMID- 6275637 TI - [Comparative studies of the efficacy of paramunity inducer PIND-AVI, mitogen PHA P and rhinopneumonitis virus on peripheral leukocytes in the horse]. PMID- 6275638 TI - Three-dimensional sequential study of the intestinal surface in experimental porcine CV 777 coronavirus enteritis. PMID- 6275639 TI - [Water-borne outbreaks of rotavirus infections]. AB - The clinico-epidemiological characteristics of group rotavirus infection observed in one settlement in March-April is presented. 173 cases of infection were registered in all age groups (children under 7 years constituted only 10.4% of the patients). In most of the apartments found to be the foci of infection only one patient was registered. The virological study of fecal samples from 24 patients showed the presence of human rotaviruses in the feces of 10 patients (41.7%). Neutralizing antibodies in titers of 1:4 to 1:64 were detected in all taken paired sera (21). THe clinical study of 53 hospitalized patients showed that in all cases the infectious process had acute beginning. In most patients the disease took mild forms. The peculiarities of the distribution of the patients in different age groups, the location of the foci of infection and its seasonal character, as well as some objective data on the presence of pollutions in the water mains, indicated that this infection could spread through water via the system of centralized water supply. PMID- 6275640 TI - [Immunogenic properties of Clostridium perfringens type A anatheta-hemolysin]. AB - Guinea-pigs were immunized with anatheta-hemolysin preparations adsorbed on aluminum hydroxide, as well as with the mixture of anatheta-hemolysin and type A Cl. perfringens toxoid, purified and concentrated. Anatheta hemolysin preparations were obtained with the use of homogeneous theta hemolysin, as well theta hemolysin of various purification degrees. As a result, antatheta hemolytic guinea-pig sera capable of neutralizing 2,000-8,000 HU of theta-hemolysin were obtained. Tests made to establish the degree of protection in the immunized guinea-pigs did not show that the animals immunized with the mixture of anatheta hemolysin and type A Cl. perfringens toxoid, purified and concentrated, had any advantages in the degree of protection over the animals immunized with the toxoid alone. But there is no doubt that this component plays a positive role under the conditions of natural gas gangrene when the hemolytic action of Cl. perfringens toxin becomes considerably pronounced. PMID- 6275641 TI - [Clinicoelectrophysiological aspects of hereditary amyotrophies]. AB - In 100 patients with the neural and spinal forms of hereditary amyotrophies the potentialities of combined electrophysiological examinations (stimulatory, general, and local electromyography) were estimated. It is shown that the method of determining the speed of the impulse transmission along the afferent and efferent fibres of the peripheral nerves and the daily fluctuations of that speed may have an additional significance in diagnosing this pathology, A comparative analysis of the data obtained showed that the most informative was combined electrophysiological examination of the patient. PMID- 6275642 TI - [Clinicoelectroneuromyographic characteristics of neuralgic amyotrophy]. AB - Clinical and genealogical examinations of 155 patients with neural amyotrophies were carried out. Use was made of general and local electromyography, determinations of the speed of impulse transmission along the efferent fibres of the peripheral nerves, and examinations of the H-reflex. The examinations revealed a great diversity of the clinical forms of the neural amyotrophies, each forms having a definite clinical picture, type of hereditary transmission, and characteristic electrophysiological changes. The electroneuromyographic examinations showed that in cases of Charcot-Marie-Tooth's classical neural amyotrophy neuritic changes were prevailing in rapidly-progressing autosomo dominant neural amyotrophies and autosomo-recessive infantile neural amyotrophy the peripheral efferent neuron was affected over its entire length. A complex examination of Roussy-Levy's syndrome revealed a pronounced dissociation between the relatively unimpaired motor function and the considerable delay of the impulse transmission along the efferent fibres of the peripheral nerves. In heterozygotic carries signs of mononeuron dysfunction were determined. PMID- 6275643 TI - [Clinicoelectroneuromyographic study of neuromuscular syndromes combined with connective tissue pathology]. AB - An electroneuromyographic examination of 132 patients with pseudohypertrophic and myosclerotic forms of myodystrophies, collagenoses, mucopolysaccharidoses, Marfan's disease, and Chernogubov-Ehlers-Danlos' disease was carried out. A relationship between the duration of the M-response potential and the state of the interstitial connective tissue of the skeletal muscles was revealed. This duration appeared to be longer in cases of proliferation, sclerosis, and fibrosis of the interstitial tissue, and shorter in cases of a deficiency of the collagen fibres and increase of their elasticity. PMID- 6275644 TI - Translation and characterization of poly(A)-lacking RNA from lupin root nodules. AB - Total polysomal RNA from yellow lupin root nodules was fractionated by double oligo(dT)-cellulose chromatography. Poly(A)-containing and poly(A)-lacking RNA fractions showed considerable messenger activity in wheat germ and rabbit reticulocyte cell-free systems. The sizing of poly(A)-lacking RNA on sucrose density gradient gives rise to separation of 14S mRNA from 22-24S mRNA species. A single polypeptide with molecular weight of 22,000 was coded for by 14S mRNA, while two polypeptides with an apparent mol. wt. of 90,000 and 87,000 were the main products of 22-24S mRNA fraction. High concentrations of unfractionated poly(A)-lacking RNA as well as the addition of poly(A) led to preferential synthesis of the 22,000 product. Preliminary results suggest the presence of m7GpppX cap structure at 5' terminus of the separated 14S and 22-24S mRNA species. This comes from the competition experiments with m7GMP and m7GTP as well as from the fact that the poly(A)-lacking RNA preparation was susceptible to methylation by methyl-transferase from vaccinia virus (methylated is the 2'-O nucleotide adjacent to 7-methylguanosine). Digestion by T1 RNAase of methylated poly(A)-lacking RNA produced two short 5'-terminal oligonucleotides 10 and 17 nucleotides in length. PMID- 6275645 TI - Hormonally stimulated adenylate cyclase and cAMP dependent protein kinase in membranes of rabbit erythroid cells separated according to density. AB - Plasma membranes were prepared after density gradient separation of erythroid cells obtained from bled animals. In a fraction enriched in young reticulocytes (lowest density), the basal and the prostaglandin stimulated adenylate cyclase were greatly augmented if compared with the membranes from unfractionated cells or from the layers of higher densities. Potentiation by GTP or soluble factor(s) of the prostaglandin stimulated adenylate cyclase was found solely in the fractions containing the youngest cells (lowest density). A very significant augmentation of both the basal and the effectors stimulated adenylate cyclase was obtained when the white blood cells and the platelets were removed by filtration through alpha-cellulose prior to density separation. A small population of probably very young reticulocytes was shown to contain a very active adenylate cyclase coupled to the hormonal receptor. Upon short time of maturation this coupling could no longer be detected. The cAMP generated in the fraction enriched in young reticulocytes increased the phosphorylation of some membrane proteins. The presence of a hormonally regulated adenylate cyclase and eventually the phosphorylation of some specific membrane proteins by the cAMP generated in situ permit to envisage possible functions of this system in young reticulocytes. PMID- 6275646 TI - [Effect of allopurinol on the liberation of free fatty acids from isolated rat and rabbit adipose tissue, stimulated by noradrenaline or adrenocortico-tropic hormone]. PMID- 6275647 TI - [Clinical study of the use of a deproteinized guar flour in the treatment of obesity]. AB - The Authors have studied an experimental product (L. G. 140/S), based on GUAR GUM, to establish its effectiveness and tolerability as a dietetic support in the treatment of obesity. The clinical research work has been conducted on 21 obese subjects (18 females and 3 males) aged 17 to 58 and treated with a hypocaloric and hypoglycemic diet. This preparation has proved useful in helping the patients to conform to the dietary restrictions and/or in normalizing their intestinal transit. No objective or subjective symptoms certainly referable to this preparation have been observed. The administration of the product has been interrupted for precautionary reasons in some cases, since it was not indispensable. Considering the good results they have obtained in terms of tolerability and agreeableness of this product, the Authors conclude that it can be considered a useful support to the restrictive dietary regimen, which is still considered - together with physical activity - the only sound and effective remedy in the treatment of obesity. PMID- 6275648 TI - Recurrence of breast cancer. A retrospective study of 569 cases in clinical stages I-III. AB - Material comprising 569 cancer patients in clinical stages I-III is presented; recurrence, factors influencing recurrence and survival after recurrence have been studied. The 10-year recurrence rate for the whole material was 52%. Frequency of recurrence correlated with the clinical stage, the axillary biopsy finding and with the malignancy grade of infiltrating ductogenic carcinoma. The incidence ratio of local recurrence and distant metastases was 1:2; the most common site of the former being the chest wall and of the latter the viscera. After diagnosis of the first local recurrence, subsequent 5- and 10-year survival rates were 42% and 24% and after diagnosis of the first distant metastasis correspondingly 15% and 13%. Survival after the detection of axillary and supraclavicular metastases was better than after the detection of chest wall recurrences. Prognosis for distant metastases was best for soft tissue metastases and poorest for visceral and multiple metastases. PMID- 6275649 TI - Carcinoma diagnosed in excised gastric ulcers. AB - Seven patients with ulcer cancer are reported. Three were peroperatively, one of them also preoperatively, suspected of malignancy and showed carcinomatous infiltration of the gastric wall, penetrating deep into the subserous layer in all cases, and lymph node infiltration in two. These patients died of their cancer, respectively 1/2, 2 and 41/2 years after operation. Four patients showed peroperative gross signs of benign gastric ulcer. In 3 of the cases carcinoma, respecting the subserous layer, was found histologically in the excised ulcer. After subsequent radical surgery in 2 and biopsy from the site of ulcer excision in 1, carcinoma was not found in the specimens. In the fourth case an excised piece of a prepyloric ulcer on the posterior wall showed superficial carcinoma. After subsequent radical operation persistent carcinoma was found in the resected specimen at the site of the biopsy. The 4 patients are alive without signs of cancer, respectively 31/2, 5, 8 and 8 years after the operation. The series demonstrates that histological examination of excised gastric ulcers may lead to the diagnosis of a carcinoma, which has limited extension and good prognosis after operative treatment. PMID- 6275650 TI - The influence of LiCL on animal halves isolated from ascidian eggs. PMID- 6275651 TI - [Usefulness of serum pseudocholinesterase isoenzymes in acute and chronic liver diseases and neoplasms (experimental and clinical study)]. AB - This paper was designed to study experimentally in rats hepatic and serum pseudocholinesterase, (CHE), and its isoenzyme activity, and also to analyze its behavior in acute hepatitis, cirrhosis and primary and secondary hepatic tumours. Five isoenzymes in rat liver homogenates and 4 to 5 in rat serum were found. In normal human serum 4 to 5 CHE-isoenzymes were recognized. Cuali and quantitative decreases in all serum CHE isoenzymes were found in all patients with severe liver disease. Isoenzyme No. 1 decreased significatively in cirrhotics, showing a double peak inscription. Isoenzyme No. 5 was elevated in the three patients with hepatoma. PMID- 6275652 TI - Influence on cerebral blood flow of infusion of sodium bicarbonate during respiratory acidosis and alkalosis in the dog. AB - In anaesthetized dogs, a mixed acid-base disturbance was induced by adding a pronounced metabolic alkaline to an established respiratory acidosis or alkalosis. Cerebral blood flow (CBF) was measured by the radioisotope washout method. In the hypocapnic dogs, the addition of metabolic alkalosis did not significantly change cerebral blood flow. In the hypercapnic dogs, the intravenous infusion of alkali led to a substantial reduction of cerebral blood flow, parallelled by a reduction of cerebrovenous oxygen tension. Acid-base analysis of cerebrospinal fluid (CSF) indicated an increased bicarbonate concentration. Hypercapnia is suggested to facilitate the passage of bicarbonate over the blood-brain barrier, leading to cerebral vasoconstriction by means of increased extravascular pH. PMID- 6275653 TI - Disopyramide anticholinergic action. AB - The effect of disopyramide on cholinergic transmission has been studied using the frog isolated abdominis rectus preparation and the guinea pig isolated vas deferens hypogastric nerve preparation. Disopyramide (2 X 10(-5) mol litre-1) reduced the response of the abdominis rectus to carbachol (1.36 X 10(-6)--4.8 X 10(-5) mol litre-1). Furthermore, disopyramide (1 X 10(-6)--3 X 10(-4) mol litre 1) produced a concentration-dependent reduction in the response to carbachol (5.46 X 10(-6) mol litre-1). The response to potassium chloride (3.34 X 10(-2) mol litre-1) was unaltered by disopyramide (1 X 10(-6)--3 X 10(-4) mol litre-1). Disopyramide produced a dose-related ganglionic blockade at concentrations greater than 2 X 10(-5) mol litre-1. Complete blockade to ganglionic transmission occurred at 3 X 10(-4) mol litre-1 disopyramide. PMID- 6275654 TI - Bell's palsy - part of a polyneuropathy? AB - Some authors have suggested that Bell's palsy should be considered a part of a polyneuritis cranialis and others, reporting Bell's palsy in patients with diabetes mellitus, proposed that it is an overt sign of a subclinical polyneuropathy. To study whether nerves other than the facial nerve were affected, the clinical findings of 112 patients with Bell's palsy were correlated with a quantitative EMG done on an asymptomatic leg muscle. The electromyographic results obtained from these leg muscles showed changes similar to those occurring in peripheral nerve lesions, and the changes correlated well with the degree of facial palsy. The results indicate that the patients with symptomatic facial dysfunction have an acute subclinical polyneuropathy. PMID- 6275655 TI - Quantitative aspects of rotavirus excretion in childhood diarrhoea. AB - The amount of rotavirus (antigen) in diarrhoeal stools was determined from serially diluted faecal specimens using a solid phase radioimmunoassay (RIA) method. The acute stage RIA titres for rotavirus antigen ranged from 1:200 to 1:200 000 with a mean titre of 1:2 936 in a group of 16 infants with age range 2 to 14 months and 1:4 703 in a group of 12 children aged 25 to 94 months. In the younger age-group the diarrhoea lasted longer (mean 7.8 days) than in the older children (mean 5.4 days) despite the fact that the quantity of rotavirus was similar. In most of the younger infants there was either no demonstrable rotavirus present at the time of cessation of diarrhoea, or the amount of virus was on the decrease. In contrast, among the older children there were cases with a high titre of rotavirus in the stools a few days after the diarrhoea had ceased. These children were considered as potential spreaders of rotavirus infection in the community. However, none of the long-term follow-up stool specimens, collected 28 to 47 days after the onset of diarrhoea, were positive for rotavirus RIA. PMID- 6275656 TI - Risk of cytomegalovirus infection in nurses and congenital infection in their offspring. AB - The risk of contracting cytomegalovirus (CMV) infection in nursing of infants and of congenital CMV infection in infants born to such nursing personnel were investigated. The investigation comprised 292 women working in paediatric clinics or day nurseries and a control group of 163 women who had no professional contact with infants. Among the women younger than 25, those who had tended infants for more than six months were significantly (p less than 0.001) more often seropositive for CMV than were those--mainly student nurses--with less than six months' infant nursing service, but ot more often than control women. At ages above 25 there was no demonstrable difference between the groups. In a separate study the occupation of 36 mothers of infants with congenital CMV infection was investigated. Compared to a control group no overrepresentation of nurses was found. All six congenitally infected infants born to nurses developed normally. PMID- 6275657 TI - A sequential study of human B lymphocyte function from birth to two years of age. AB - The immunoglobulin synthesizing capacity of individual lymphocytes from newborns and infants aged 2, 6, 12 and 24 months respectively has been studied. The technique has been to expose purified lymphocytes to a T lymphocyte dependent (pokeweed mitogen) or a T lymphocyte independent (Epstein-Barr virus) activator of B lymphocytes. Activation has been measured as immunoglobulin secretion of individual B lymphocytes using a hemolytic plaque assay. B lymphocytes from newborns can be made to synthesize IgM at adult levels, but not IgG and IgA. Within 24 months from birth the secretion of IgG has reached adult capacity whereas IgA formation is still diminished. Lymphocytes synthesizing IgG subclasses appear at different times insofar as IgG1 and IgG3 are well demonstrable within 12 months from birth whereas IgG2 and IgG4 has not at all reached adult levels even 24 months after birth. The T lymphocyte dependent activator (pokeweed mitogen) fails to induce immunoglobulin synthesis in peripheral blood lymphocytes from newborns because of a defect helper T cell function. Such T cell capacity appears at the age of 6 months. The data unequivocally demonstrate restricted but definite B lymphocyte functional capacity already at birth and a gradual but partial acquisition of adult competence until the age of 2 years. PMID- 6275658 TI - [Effect of alcohols on synaptic stimulation transmission in the isolated sympathetic ganglion of the frog]. PMID- 6275659 TI - Characteristics and behavior of plasma membranes. PMID- 6275660 TI - The role of dietary fiber in lipid metabolism. PMID- 6275661 TI - Effects of hypolipidemic drugs on bile acid metabolism in man. PMID- 6275662 TI - Cholesterol metabolism by ovarian tissue. PMID- 6275663 TI - Cranial neural crest cells: inhibition of differentiation in culture. PMID- 6275664 TI - Receptor interactions of nerve growth factor. PMID- 6275665 TI - Cellular interactions in Schwann cell development. PMID- 6275666 TI - Refractory period measurement in the clinical domain. PMID- 6275667 TI - Hyperexcitability of pathologically myelinated axons and positive symptoms in multiple sclerosis. PMID- 6275668 TI - Action potential electrogenesis in mammalian central axons. PMID- 6275669 TI - Ionic and gating currents in mammalian myelinated nerve. PMID- 6275670 TI - Distribution of sodium and potassium channels in mammalian myelinated nerve. PMID- 6275671 TI - Pharmacological properties of sodium channels in nerve membranes. PMID- 6275672 TI - Isolation of sodium channels from excitable tissues. PMID- 6275673 TI - A review of membrane structure with perspectives on certain transmembrane channels. PMID- 6275674 TI - Approaches to the development of pharmacological interventions in multiple sclerosis. PMID- 6275675 TI - Electrophysiology of demyelinating diseases: future directions and questions. PMID- 6275676 TI - Effects on arterial receptors of ergot derivatives used in migraine. PMID- 6275677 TI - Impairment of cerebral serotonin and energy metabolism during ischemia: relevance to migraine. PMID- 6275679 TI - Serotonin and cyclic nucleotides in migraine. PMID- 6275678 TI - Monoamine sensitivity of smooth muscle in vivo in nociception disorders. AB - A significant degree of supersensitivity to 5-HT and DA was detected when carrying out the computerized venotest on migraine patients during an attack. A similar supersensitivity was observed during morphine abstinence and naloxone precipitated withdrawal in addicts. Mild abstinence after slight and short morphine treatment provoked monoamine supersensitivity in volunteers. In these conditions, the administration of morphine inhibited the 5-HT and DA supersensitivity. In spontaneous central panalgesia, monoamine supersensitivity is detectable, as well as in panalgesia induced in headache sufferers by means of PCPA 5-HT deprivation. By means of the venotest, the ergot derivatives were confirmed as being partial 5-HT agonists. These drugs can also carry out their therapeutic activity by potentiating 5-HT at a central level in 5-HT-deficient neurons. The presence of opiate receptors in the human vein is stressed. The high supersensitivity of the venous smooth muscle to 5-HT and DA both in headache and systemic pain sufferers and during morphine withdrawal suggests a pathophysiological analogy between these conditions. PMID- 6275680 TI - Endorphins and modulation of pain. PMID- 6275681 TI - Natural opioids in migraine. PMID- 6275682 TI - Endorphins in the pathogenesis of headache. PMID- 6275683 TI - Effect of antimigraine drugs on nonopioid analgesia. PMID- 6275684 TI - New enteric viruses in the dog. PMID- 6275685 TI - Motor functions of the intestine. PMID- 6275686 TI - Persistent viral infections of food animals: their relevance to the international movement of livestock and germ plasm. PMID- 6275687 TI - Mechanisms of hemostasis and therapy of thrombosis: new concepts based on the metabolism of arachidonic acid by platelets and endothelial cells. PMID- 6275688 TI - [Types of ocular attacks and lysosomal enzymes in Behcet's disease (author's transl)]. PMID- 6275689 TI - [Experimental endophthalmitis induced by herpes simplex virus. 2. Intracarotid inoculation (author's transl)]. PMID- 6275690 TI - [Thin-layer chromatography separation of L-ascorbic acid from its oxidation products (author's transl)]. PMID- 6275691 TI - PIPIDA scintigraphy for cholecystitis: false positives in alcoholism and total parenteral nutrition. AB - A review of gallbladder scintigraphy in patients with potentially compromised hepatobiliary function revealed two groups in whom cholecystitis might be mistakenly diagnosed. In 200 consecutive hospitalized patients studied with technetium-99m-PIPIDA for acute cholecystitis or cholestasis, there were 41 alcoholics and 17 patients on total parenteral nutrition. In 60% of the alcoholics and 92% of those on parenteral nutrition, absent or delayed visualization of the gallbladder occurred without physical or clinical evidence of cholecystitis. A cholecystagogue, sincalide, did not prevent the false positive features which presumably are due to altered bile flow kinetics related to alcoholism and parenteral nutrition. Four patients on parenteral nutrition undergoing cholecystectomy for suspected cholecystitis had normal gallbladders filled with jellylike viscous thick bile. A positive (nonvisualized or delayed visualized) gallbladder PIPIDA scintigram in these two populations should not be interpreted as indicating a need for cholecystectomy. PMID- 6275692 TI - Breast parenchymal patterns: analysis of 332 incident breast carcinomas. AB - A detailed analysis of 332 incident breast carcinomas is presented. The study results indicate that there is validity to the breast parenchymal patterns as an indication of risk for developing breast cancer and that risk is independent of length of follow-up and age of patient. All cases were drawn from a referral-type practice at Hutzel Hospital, Detroit, MI. The cases were compared to controls for computation of relative risk up to and beyond a follow-up period of 48 months. They were also compared to a consecutive series of prevalent breast carcinomas seen at Hutzel Hospital during the 6 years 1975-1980. The extent of disease in the cases described in this report is somewhat more favorable than in a consecutive series of prevalent cancers operated on a Hutzel Hospital. Most breast cancers occur in the P2 breast, a smaller number in the DY, and only 12% in the combined N1P1 type. The conclusions of other investigators regarding parenchymal patterns as risk indicators are reviewed. Particular attention was paid to comparing the length of follow-up and age of patient. There was no diminution of proportions of cases in the P2/N1-P1 or DY/N1-P1 after a 48 month follow-up. There was a significant diminution in the proportion of cases falling into DY/N1-P1 after age 50, and less so in P2/N1-P1. Furthermore, relative risk was unaffected by time or age. PMID- 6275693 TI - Abdominal scintigraphy for ectopic gastric mucosa: a retrospective analysis of 143 studies. AB - A retrospective analysis of 143 studies of abdominal scintigraphy with technetium 99m pertechnetate for the diagnosis of ectopic gastric mucosa (usually located in Meckel diverticulum) was undertaken. The rectilinear scanner was used for 57 studies and the gamma camera for 86. The final diagnosis, at laparotomy or after clinical and laboratory investigation with long-term clinical follow-up, was correlated to the interpretation of the scintigrams. Of 57 rectilinear scans, five were true positive, 46 true negative, two false positive, and four false negative. Of the 86 camera studies 10 were true positive, 74 true negative, one false negative, and one patient showed large bowel activity (positive atypical study). The results of this analysis indicate the value of the properly performed and interpreted abdominal pertechnetate scintigraphy in the diagnosis of (bleeding) ectopic gastric mucosa; it was also shown that the sensitivity and the specificity of this test have improved since the introduction of the gamma camera and the standardization of the procedure. PMID- 6275694 TI - Nuclear magnetic resonance imaging of the liver: initial experience. AB - Nuclear magnetic resonance (NMR) scans of the liver were obtained in 12 normal volunteers and 32 patients using a whole-body machine developed by Thorn-EMI Ltd., and the results were compared with x-ray computed tomography (CT). Two types of NMR scan, saturation-recovery and inversion-recovery, were performed in order to obtain values for the spin-lattice relaxation time, T1. Although the saturation-recovery scans show little soft-tissue detail, the inversion-recovery scans demonstrated the interlobar fissure, hepatic veins, portal veins, bile ducts, and gallbladder. In comparison with CT (Siemens Somatom 2), both types of NMR scan showed some blurring due to respiratory movement but much less linear artifact across the liver from the air-fluid interface in the stomach. Focal disease within the liver was demonstrated by both CT and NMR, although an area of focal atrophy and another of hepatic infarction were only recognized with NMR. In diffuse disease the pattern varied. In steatosis CT was virtually diagnostic, while NMR showed no specific features. In hemochromatosis, hepatitis, eight cases of cirrhosis, and one of Wilson disease, both techniques showed abnormalities of varying specificity. In two cases of cirrhosis and one of primary biliary cirrhosis, only the NMR scan was abnormal. Nuclear magnetic resonance images are now sufficiently anatomically detailed to permit serious comparisons with technically advanced computed tomography. The information revealed is fundamentally different and can be expected to have some diagnostic utility. PMID- 6275695 TI - Disseminated mucormycosis in a case of metastatic carcinoma. AB - A 50-year-old white woman who underwent bilateral mastectomy for fibrocystic disease of the breast in 1974 and laminectomy for lumbar disc fusion in 1979, was diagnosed early in 1980 at Roswell park Memorial Institute as a case of metastatic anaplastic carcinoma. She was started on platinum combination therapy, but in the course of her treatment she became increasingly jaundiced, then semi comatose and died suddenly two months after the diagnosis of carcinoma. Histopathologic examinations revealed undifferentiated small-cell carcinoma in the lung, liver, lymph nodes, and bone. Mucormycosis, characterized by the presence of broad coenocytic hyphae associated with thrombosis, infarction, and necrosis, was unexpectedly discovered at postmortem examination in the brain, lung, and heart. The etiologic agent, Absidia corymbifera, was isolated from a lung specimen and was identified by its morphologic characteristics and by the formation of zygospores in crosses with a compatible tester strain of the mating type (-). PMID- 6275696 TI - Collaborative calibration of the U. S. National and the College of American Pathologists reference preparations for specific serum proteins. PMID- 6275697 TI - Russell bodies: a light and electron microscopic immunoperoxidase study. AB - Russell bodies have been previously regarded as aggregates of immunoglobulin. Light and electron microscopic immunoperoxidase studies show no detectable immunoglobulin determinants in the Russell body cores. Denaturation of antigens during tissue preparation appears to be an unlikely explanation, since immunoglobulins in the cytoplasm of plasma cells are clearly demonstrated. The presence of immunoglobulins on the surface of small intracellular Russell bodies may represent the immunoglobulin determinants in the surrounding rough endoplasmic reticulum. It seems likely that Russell bodies contain non immunoglobulin molecules, by-products of immunoglobulin synthesis, or some altered form of immunoglobulins that no longer can be recognized by the anti immunoglobulin antibody. The non-uniform dye staining pattern of Russell bodies further suggests that Russell bodies may be heterogenous in nature. PMID- 6275698 TI - Sarcoidosis. PMID- 6275699 TI - Infantile vitamin D-resistant rickets associated with total parenteral nutrition. PMID- 6275700 TI - Stability of an extemporaneous formulation of injectable cholecalciferol. AB - THe stability of an extemporaneous formulation of injectable cholecalciferol 40,000 IU/ml was assessed. A preparation of cholecalciferol 40,000 IU/ml in a solution of propylene glycol with ethanol 10% was prepared. The solution was stored at 4 degrees C in 2-ml sterile glass vials protected from light. Cholecalciferol content was measured initially and periodically for 199 days using a high-pressure liquid chromatography technique. A 7.3% loss of cholecalciferol potency from baseline was observed over the 199-day period. In contrast to currently available preparations, this extemporaneous formulation of injectable cholecalciferol potentially offers a versatile dosage form for therapeutic or prophylactic therapy of vitamin D deficiency. It was easily prepared and retained more than 90% potency of cholecalciferol for at least six months. The addition of this formulation to large-volume parenteral or parenteral nutrition solutions is not recommended, however, because compatibility data are not available. In addition, the biological activity of the preparation has not been assessed. PMID- 6275701 TI - Calculation of sodium bicarbonate requirement in metabolic acidosis. AB - Despite the host of complications which may be associated with intravenous sodium bicarbonate infusion, the use of this agent is a frequent necessity in patients with metabolic acidosis. No satisfactory formula for calculating bicarbonate dose had previously been described, although such an approach might be expected to reduce the incidence of these complications. The authors have devised a simple formula for bedside calculation of bicarbonate requirement in metabolic acidosis, designed to elevate th pH to the region about 7.30, and report their experience with the use of this formula in 13 instances. In all but one, the post-infusion pH was between 7.25 and 7.37, with a mean of 7.30 +/- 0.04 and no instances of serious overtitration. It is concluded that the formula is useful as a pragmatic aid in the management of patients with metabolic acidosis. PMID- 6275702 TI - Infrequent elevation of plasma ACTH in patients with bronchogenic carcinoma. AB - Older methods for radioimmunoassay of ACTH in human plasma have detected a high frequency of plasma ACTH elevation in patients with lung cancer, even in the absence of Cushing's syndrome. It now appears evident that these assay systems measure biologically inactive forms and fragments of ACTH. Since ectopic Cushing's syndrome may be clinically atypical, a radioimmunoassay for ACTH that correlates with the clinical state would be of value. Accordingly, circulating plasma corticotropin (ACTH) levels were measured in 39 consecutive patients with biopsy-proven, untreated lung cancer, all without evidence of Cushing's syndrome. In only one instance was ACTH found to be elevated by the method of assay used. Two interpretations are justified: (1) elevated plasma ACTH levels are infrequent in patients with lung cancer, or (2) ACTH measurements vary according to the assay system used. PMID- 6275703 TI - Cushing's syndrome due to ectopic ACTH production by a nasal paraganglioma. AB - A patient is described with Cushingoid features and a tumor of the paranasal sinuses. Laboratory studies revealed hyperglycemia, hypokalemia, alkalosis, hypercortisolemia, and elevated serum ACTH levels. The cortisol level increased briskly in response to parenteral ACTH, but was only partially suppressed by dexamethasone. Microscopically, the tumor was found to be a paraganglioma containing many neurosecretory granules.. Treatment with methyrapone, o,p'DDD, and irradiation of the tumor resulted in a lasting remission. It is concluded that this unique patient suffered from Cushing's syndrome due to ectopic ACTH production by a paraganglioma of the paranasal sinuses. PMID- 6275704 TI - Chemotherapy-related conversion of tumor. PMID- 6275705 TI - Roentgenologic manifestations of pulmonary metastases in choriocarcinoma and invasive mole. AB - From 1949 to 1975, a total of 3,915 chest films were taken for 429 cases of choriocarcinoma and 441 cases of invasive mole. The incidences of pulmonary metastases were 85.1% and 65.0%, respectively. The various forms of pulmonary metastases were studied and correlated with clinical symptoms and pathologic changes. In order to follow up the progression or regression of the various forms, serial chest films were taken at intervals of 10 to 14 days in 27 cases. In eight cases postmortem pulmonary arteriograms were obtained on the autopsied lung specimens for the study of vascular changes of metastatic lesions. Pathologic examinations and pulmonary arteriography were also done on the surgically resected lung specimens. As a result of this study, a relative comprehensive knowledge about the nature and development of the various forms of metastatic shadows has been deduced. It is rational to say that the various forms seen on the chest films represent only the various evolutionary changes of the same lesion. PMID- 6275706 TI - Psychological changes effected by estrogen-progestogen and clonidine treatment in climacteric women. AB - The psychological responses of 23 symptomatic climacteric women were analyzed to compare the effects of opposed estrogen therapy, i.e., conjugated equine estrogen (Premarin) and medrogestone (Colpro), with those of clonidine (Dixarit). Ten asymptomatic postmenopausal women constituted a control group. Statistical analysis of the data of this randomized double-blind prospective study showed that the hormone-treated group experienced significant improvement in various measurements of depression and anxiety whereas the clonidine-treated group did not. Ratings based on subjective reports by the patients support these findings. Various inverse and positive associations between the psychological variables are presented. PMID- 6275707 TI - Peritoneal macrophage alterations caused by naturally occurring mouse hepatitis virus. AB - During routine harvest of murine resident peritoneal cells for macrophage function assays the authors recently noted that mice showed a 3-4-fold spontaneous increase in number of peritoneal cells within 1 week of being placed in one of their animal rooms. While the mice appeared clinically normal, the collected macrophages had highly convoluted membranes, showed enhanced spontaneous tumor cell killing, and showed increased erythrophagocytosis. Histopathologic findings included mild peritonitis and occasional foci of individual hepatocyte necrosis. The results of routine murine serologic studies and bacterial cultures of the peritoneal cavity were negative. Immunosuppressed mice placed in this room showed severe hepatic necrosis within 4 days, and ultrastructural particles characteristic of corona virus could be demonstrated in the necrotic foci. Mouse hepatitis virus (MHV) was isolated from these livers. Untreated mice showed positive MHV titers, as detected by the enzyme-linked immunoabsorbent assay (ELISA) after 21 days in the room. This episode demonstrates that MHV have profound effects on macrophage parameters while causing few clinical signs or histopathologic alterations. Secondly, the complement fixation assay for MHV as included in routine viral screens appears relatively insensitive for detecting outbreaks of MHV. PMID- 6275708 TI - Ultrastructural immunohistochemical localization of virus in acute and chronic demyelinating Theiler's virus infection. AB - Mice experimentally infected with Theiler's murine encephalomyelitis virus (TMEV) develop a persistent infection of the central nervous system (CNS). The most striking feature of this infection is the occurrence of inflammatory primary demyelination in the spinal cord white matter. The pathogenesis of myelin degeneration in this model has not been clarified, but morphologic and immunologic data suggest that the host immune response plays a major role in the production of myelin injury. Because of low virus titers in infected adult mice and of the small size of TMEV, virus particles have never been observed in this demyelinating model. Yet elucidation of the types of cells in the CNS supporting virus replication would be important for a better understanding of both virus persistence and virus-induced demyelinating pathology. The present paper is a sequential study of the localization of TMEV in the spinal cord in infected mice by ultrastructural immunohistochemical techniques. Results indicate that virus replication is mainly in neurons during the acute phase of the disease, while in the chronic phase viral inclusions are mainly found in macrophages in and around demyelinating lesions. Other cells are also infected, but to a lesser degree. In the neuronal system both axoplasmic and dendritic flow appear to facilitate the spread of virus in the CNS. In macrophages, the presence of virus particles and the association of virus with altered components of the cytoskeleton support active virus production rather than simple internalization. The macrophage appears to play an important role in both the establishment of virus persistence and in the process of demyelination in this animal model. PMID- 6275709 TI - A quantitative ultrastructural study of peripheral blood lymphocytes containing parallel tubular arrays in Epstein-Barr virus and cytomegalovirus mononucleosis. AB - In the normal peripheral circulation there exists a subpopulation of lymphocytes that is ultrastructurally distinct. This lymphocyte is identified with the electron microscope by the presence of cytoplasmic microtubulelike inclusions called parallel tubular arrays (PTAs) and contains Fc-receptors for cytophilic antibody. In this study, lymphocytes containing PTAs (PTA-lymphocytes) were quantitated from serial peripheral blood specimens obtained from two patients with Epstein-Barr virus (EBV) mononucleosis and two patients with cytomegalovirus (CMV) mononucleosis. These data were then correlated with the clinical state of the patient. It was determined that both the percentage and absolute number of PTA-lymphocytes were highest during the acute phase of the illness. In follow-up specimens, three of the four patients' absolute lymphocyte count fell to within normal limits before the absolute PTA-lymphocyte count. In one patient, the absolute PTA-lymphocyte count was significantly elevated 13 months after the initial clinic visit. Although the PTA-lymphocyte count was highest during the acute phase of the illness, there was no consistent correlation with the clinical state of the patient during follow-up. The estimation of absolute PTA-lymphocyte counts was determined to be valid after a morphometric analysis of the cellular areas occupied by PTAs during the acute and convalescent phases of the disease revealed no statistical differences. Electron microscopy was also performed on the peripheral blood of a patient with syphilis. Although a hematologic workup of this patient during the acute phase of his illness revealed a large number of atypical lymphocytes, electron-microscopic examination of the same specimen revealed both a normal number and a normal percentage of PTA-lymphocytes. The immunologic role of this ultrastructurally distinct third population (non-T, non B) of lymphocytes, or "killer cells," in the course of infectious mononucleosis is discussed. PMID- 6275710 TI - Alterations in the proximal nephron of beige mice with the Chediak-Higashi syndrome. AB - The proximal nephron of C57 beige mice with a genetic defect analagous to the Chediak-Higashi syndrome (CHS) has been compared with that of normal C57 black mice. The concanavalin A-horseradish peroxidase (Con A-HRP) technique stained the brush border of the proximal straight tubule heavily in black mice and weakly in beige mice. In beige mice this method stained the brush border of the proximal convoluted tubules weakly and the brush border of the proximal straight tubules only negligibly. Periodic acid-Schiff staining showed no such difference between beige and black mice but revealed an increase distally in the size of the CHS inclusions in the proximal straight tubule of beige mice. Immunostaining visualized abundant lysozyme in the first portion of the proximal nephron but none in the more distal segments of beige and black mice alike. At the ultrastructural level, the proximal convoluted tubules of black mice contained two morphologic types of heterophagosomes, which apparently differed in accord with the stage of their development. Proximal straight tubules contained morphologically different heterophagic bodies. The mature stages of these heterophagosomes were greatly enlarged in CHS mice. With the periodic acid thiocarbohydrazide-silver proteinate (PA-T-SP) method for localizing glycoprotein ultrastructurally, the microvillar brush border, apical invaginations of the plasmalemma, Golgi cisternae, and lysosomal inclusions stained selectively in the proximal nephron in both strains. The proximal straight nephron of beige mice after staining with the PA-T-SP method appeared depleted of the strongly reactive apical invaginations in some areas, particularly where large heterophagosomes bordered the apical plasmalemma. The enlarged secondary lysosomes of heterophagic origin in beige mice varied in showing both diffuse and focal PA-T-SP reactivity. Lysosomal acid phosphatase activity appeared decreased, and peroxisomes were normal in size but increased in number in the proximal nephron of beige mice. PMID- 6275711 TI - Blood groups, serum proteins, and red cell enzymes in the Nganasans (Tavghi) reindeer hunters from Taimir Peninsula. AB - The Nganasans are made up of two recently tribal populations. These, the Avam and Vadey, were established in the seventeenth century from small reindeer hunting bands, themselves apparently descended from the Yukaghir. Data on 13 blood systems have been described for the first time in the Vadey Nganasans, and the results compared with those previously reported for the two Avam subgroups. As a whole, the Nganasans are characterized by low frequency of B blood group, high frequencies of Ns, cDE, Fy(a), Hp(2), absence of A2, P(c), K, and apparently an absence of cde alleles or haplotypes. Measurement of intrapopulation heterogeneity reveals significant divergence among the two Avam subdivisions (chi 2/16=57.59; P less than 0.001), as well as between the total Avam and Vadey (chi 2/17=79.31; P less than 0.001). Founder principle, and local genetic drift, are believed to account for the greater difference between the Avam and Vadey subgroups than that observed between the two Avam populations. The Nganasans of the Taimir Peninsula appear to be the last group of reindeer hunters remaining in Northern Siberia. For ages they have lived in relative isolation, and therefore are the least touched genetically, either by surrounding herding groups originating in Southern Siberia, or by recent Caucasian admixture. PMID- 6275712 TI - Hypothalamic control of endocrine thermogenesis. AB - Hypothalamic and ruminal cooling raised serum thyrotropin (TSH), adrenocorticotropin (ACTH), norepinephrine (NE), and glucose in conscious goats in 20 degree C ambient temperature. Cooling of the preoptic anterior hypothalamus (POAH) for 2 h initially evoked shivering and vasoconstriction, leading to 1.5 degree C rise in rectal temperature (Tr). Pituitary-thyroid activation by POAH cooling was shown by peak rises in TSH of 60% at 40 min, in triiodothyronine (T3) of 54% at 80 min, and in thyroxine (T4) of 40% at 140 min. At 60 min, ACTH and NE peaked at 57 and 65%, respectively. TSH, ACTH, and NE declined during the 2nd h of POAH cooling as Tr plateaued; when POAH cooling was stopped, these hormones fell below basal level as vasodilation and panting restored Tr to normal. In contrast to the core hyperthermia evoked by POAH cooling, ruminal intubation with O degree C water (1 liter/10 kg) led to general hypothermia, Tpoah and Tr falling 1.6 degree C at 40 min. Pituitary-thyroid responses were less but ACTH and NE more, compared with POAH cooling. TSH peaked at 37% at 20 min, T3 at 55% at 60 min, and T4 at 18% at 200 min. ACTH peaked at 250% at 30 min and NE at 120% at 20 min. Thermosensitive neurons in the POAH seem to mediate more sensitive and complete control over TSH than over ACTH, or NE release, whereas extrahypothalamic core thermosensitivity (e.g., brain stem, spinal cord, abdomen) may influence ACTH and NE more than TSH release. PMID- 6275713 TI - Thyroid hormone-sensitive brown adipose tissue respiration in the newborn rabbit. AB - The effects of thyroid hormone treatment on brown adipose tissue (BAT) and liver metabolism were assessed by measuring oxygen consumption, sodium-potassium adenosine triphosphatase (Na-K-ATPase), and mitochondrial alpha-glycerophosphate dehydrogenase (alpha-GPD) activities in tissues from triiodothyronine- (T3) and vehicle-injected (for 3 days) newborn and adult rabbits. In the newborns, basal BAT cellular respiration was increased [mean (%/- SE) = 119 +/- 18 vs. 65 +/- 4 microliter O2/10(6) cells-1 . h in controls (P less than 0.005)], whereas hepatic respiration was unchanged. Ouabain had no effect on basal BAT cellular respiration, but suppressed hepatic respiration by 30% in both newborn groups. T3 treatment had no effect on NE- (10(-6) M) stimulated BAT respiration, whereas adult hepatic respiration was increased almost twofold. alpha-GPD activities were increased in both newborn BAT and adult liver but not in newborn liver. Na-K ATPase activity was significantly increased only in newborn liver. In conclusion, 1) both BAT and liver are thyroid-hormone sensitive in the newborn rabbit, but the responses to T3 treatment are different in the two tissues; 2) the failure to stimulate both hepatic alpha-GPD and respiration in the newborn appears to be a developmental phenomenon characteristic of the rabbit; 3) thyroid hormones have little effect on sodium transport-dependent respiration in either BAT of liver in the newborn rabbit. PMID- 6275714 TI - Mineralcorticoid receptors along the nephron: [3H]aldosterone binding in rabbit tubules. AB - To identify the site of mineralocorticoid action along the nephron, we measured the specific binding of [3H]aldosterone to nephron segments microdissected from aldosterone-deficient rabbits. Specific binding was defined as the difference between binding measured in the absence or in the presence of 2,000-fold excess of unlabeled hormone (in 10(-18) mol X cm tubule length-1 +/- SE). High specific binding capacity was found in the branched collecting tubule (108 +/- 4), the cortical collecting tubule (119 +/- 9), and the outer medullary collecting tubule (115 +/- 16), whereas specific binding was negligible in the proximal convoluted tubule (8 +/- 9), pars recta (2 +/- 6), medullary thick ascending limb (4 +/- 6), cortical thick ascending limb (6 +/- 2), and distal convoluted tubule (6 +/- 6). In cortical collecting tubules, Scatchard analysis of the specific [3H]aldosterone binding indicated a dissociation constant (KD) of 2.2 X 10(-9) M and a maximum number of binding sites of 157 X 10(-18) mol X cm tubule length-1. The steroid specificity was assessed from the competition of various steroids for [3H]aldosterone binding sites. Receptors from the cortical collecting tubule revealed the following sequence of affinities: aldosterone greater than DOCA greater than spironolactone greater than dexamethasone greater than 5 alpha dihydrotestosterone = progesterone = 17 beta-estradiol, indicating that the binding sites in the collecting tubule are mineralocorticoid receptors. These results demonstrate significant [3H]aldosterone binding to receptors of high affinity and mineralocorticoid specificity only in the collecting tubule and suggest that this nephron segment is the target site of mineralocorticoid action in the rabbit kidney. PMID- 6275715 TI - Multiple sites for interaction of prostaglandin and vasopressin in toad urinary bladder. AB - The interaction of vasopressin with prostaglandins were examined in the toad bladder by determining water flows, cAMP levels, and cAMP-dependent protein kinase activity. Both water flow and activation of cAMP-kinase in response to vasopressin were enhanced after prostaglandin inhibition, consistent with inhibition of vasopressin-induced cAMP generation by endogenous prostaglandins. On the other hand exogeneous PGE stimulated cAMP generation. PGE1 (10(-7) M) alone did not increase water flow but activated kinase more than vasopressin only. Addition of PGE1 (10(-7) M) and vasopressin inhibited water flow as compared with vasopressin along but increased the kinase ratio above that with vasopressin only. PGE2 (10(-5) M) increased the cAMP content and kinase ratio even more than vasopressin but again resulted in no water flow. Addition of vasopressin and PGE2 (10(-5) M) increased water flow but did not alter cAMP content or the kinase ratio compared with PGE2 alone. Similar results were obtained with PGE1. Accordingly, prostaglandin dissociates cAMP levels and kinase ratio from the hydroosmotic response, suggesting that PGE2 inhibits steps distal to cAMP. Consistent with this, in bladders pretreated with naproxen or meclofenamate, PGE2 (10(-8) to 10(-6) M) inhibited the response to submaximal doses of cAMP (5 mM) or 8-bromo-cAMP (0.03 mM). Furthermore, pretreatment with naproxen significantly enhanced the response to cAMP (5 mM). These studies provide evidence for vasopressin-PGE interaction at the site of cAMP generation and also at a step(s) unrelated to cAMP generation. PMID- 6275716 TI - Effects of vasopressin and urea on Ca2+-calmodulin-dependent renal prostaglandin E. AB - Vasopressin (AVP stimulated immunoreactive E (iPGE) synthesis and the release of [3H]arachidonate (AA) from prelabeled slices of rat inner medulla (IM) in the presence but not in the absence of Ca2+ (plus 2 mM EGTA). Urea (700-1,200 mosM) inhibited these actions. By contrast Ca2+ deprivation or urea did not suppress AVP-induced increases in cAMP or stimulation of iPGE by exogenous AA. At 10-50 microM, trifluoperazine (TFP) suppressed AVP-induced increases in [3H]AA release and iPGE accumulation, but not increases in cAMP. Basal acyl hydrolase activity (AH) of 2,000 g particulate fractions of IM was suppressed by EGTA. Ca2+, but not Mg2+ or AVP, restored AH to levels observed in the absence of EGTA. Ca2+-induced increases in particulate AH were inhibited by urea (700-1,200 mosM) or TFP (10-50 microM), but not 1,000 mosM NaCl. Partial depletion of particulate calmodulin like activity suppressed Ca2+-induced increases in AH. The latter was restored with 1 microM purified exogenous calmodulin but not troponin C. The results demonstrate that AVP stimulation of AA and PGE release in IM are Ca2+-dependent processes suppressed by urea. They also suggest a role for Ca2+-calmodulin dependent AH in the control of PG synthesis in IM. PMID- 6275717 TI - Intracellular potassium activity in the rabbit proximal straight tubule. AB - Double-barreled liquid ion-exchanger microelectrodes were used to measure basolateral membrane potential (VBL) and intracellular potassium activity (aiK) in superficial proximal straight tubules (sPST) of the rabbit perfused in vitro. The mean +/- SE (number of cells in parentheses) value of VBL was -37.8 +/- 2.49 (20) vM and aiK was 48.6 +/- 2.27 (20) mM. The calculated Nernst equilibrium potential (EK) across the basolateral membrane was -68 mV. Lowering both potassium concentration to 0.1 mM reversibly decreased both VBL and aiK to -12.2 +/- 1.21 (19) mV and 11.3 +/- 1.29 (19) mM, respectively. Bath ouabain (10(-5) resulted in similar changes. These results demonstrate that intracellular potassium is actively accumulated in sPST perfused in vitro and that accumulation results primarily from Na-K-ATPase activity in the basolateral membrane. During recovery from low K bath, the temporal relationship between VBL and aiK and the effects of ouabain and high K bath on recovery are used to demonstrate directly electrogenic pumping. Lowering bath pH to 6.7 (HCO-3 = 5 mM) and the presence of 0.5 mM BaCl2 in the bath resulted in a large and rapid depolarization of VBL with little or no change in aiK. These results suggest that the response of VBL to both maneuvers is caused by a decrease in potassium permeability of the basolateral membrane. PMID- 6275718 TI - Sodium-coupled nonelectrolyte transport across epithelia: emerging concepts and directions. AB - Since the proposal of the sodium-gradient hypothesis, research efforts have focused on two major areas: 1) establishing the thermodynamic efficacy of the electrochemical gradient for sodium as a driving force for uphill solute accumulation, and 2) determining the mechanism of energy coupling in cotransport systems. On the whole it is now reasonably well established that the Na+ electrochemical gradient can indeed energize the uphill accumulation of various nonelectrolytes in epithelia. This conclusion is substantiated by electrophysiological studies on intact epithelia using both conventional and Na+ selective microelectrodes and has been repeatedly verified in studies using brush border membrane vesicles. One important additional concept that has emerged from these studies is that the overall transport process is intimately associated with, and perhaps controlled by, the electrical properties of leaky epithelia. The mechanism of energy coupling, as deduced from kinetic studies and modeling, has proven to be more elusive. Interpretations of earlier kinetic measurements on intact epithelia are complicated by unstirred layer effects and varying electrochemical Na+ gradients. Even more recent studies using brush-border membrane vesicles have failed to provide a unified mechanism, even for Na+ glucose transport, because of the various methods used to extract kinetic parameters and the variety of assumptions underlying different kinetic models. The common assumption that a translocation step rate limits the coupled entry process has been recently challenged, leading to novel proposals that are not based on mobile carriers. Future mechanistic studies will undoubtedly rely on recent advances in the isolation and purification of Na+-dependent proteins and their subsequent reconstitution into well-defined artificial membranes. PMID- 6275719 TI - Alkaline secretion by amphibian duodenum. III. Effect of DBcAMP, theophylline, and prostaglandins. PMID- 6275720 TI - ECT and diabetes mellitus. PMID- 6275721 TI - A study of intestinal protozoa including non-pathogenic Entamoeba histolytica from patients in a group of mental hospitals. AB - Twenty-five per cent of 174 patients residing in three mental hospitals were found to be infected with protozoa. Seven per cent of the patients were infected with Entameba histolytica. Isoenzyme characterization of the E. histolytica grown in culture showed that there were no pathogenic zymodemes of this organism in any of the specimens tested. (Am J Public Health 1982;72:178-180.) PMID- 6275722 TI - Rhabdomyogenesis in renal neoplasia of childhood. AB - Of 220 consecutive primary renal tumors of childhood, 17 contained substantial amounts of histologically identifiable striated muscle cells (over 10% of sampled tumor parenchyma). These tumors could be further subclassified into two groups: Wilms' tumors with "massive" rhabdomyogenesis (one-third or more of the tumor parenchyma composed of muscle), and Wilms' tumors with "moderate" rhabdomyogenesis (10-30% muscle composition. The former tumors were invariably seen in young children, under 4 years of age; often the patients were infants 1 year of age, or younger, and more than half of the patients in this group had bilateral tumors. Bilaterality was not seen in patients harboring tumors with "moderate" rhabdomyogenesis, whom in addition, were older children. In both groups, there was a tendency for polypoid intrapelvic growth. All but one of the tumors described in this report were classified as Wilms' tumor; the single exception was considered be be a primary rhabdomyosarcoma of the kidney. Patients with congenital malformations related to Wilms' tumor (one aniridia, one hemihypertrophy) were seen only in the group with"massive" rhabdomyogenesis. However, anatomical lesions consistent with neoplastic multifocal origin were present in both groups. Thus, our findings indicate a definite correlation between extensive rhabdomyogenesis and clinical behavior. This relation is expressed in patterns of age distribution, bilaterality and manner of growth, which are sufficiently consistent to individualize this histologic variant as a cytodifferentiated form of nephroblastoma. PMID- 6275723 TI - Alpha-1-antitrypsin in a malignant mixed mesodermal tumor of the ovary. AB - A malignant mixed mesodermal tumor of the ovary was found to contain multitudinous clusters of eosinophilic hyaline globules in both its epithelial and mesenchymal components. These periodic acid-Schiff-positive, diastase resistant globules revealed typical positive ring-like staining for alpha-1 antitrypsin (AAT) using an indirect immunoperoxidase technique. Similar findings have previously been reported in certain germ cell and liver tumors and hepatocytes of patients bearing the Z-allele for AAT. The globules also stained with antisera against kappa and lambda immunoglobulin light chains, indicating the presence of surface immunoglobulin. In the electron microscope, the inclusions appeared granular and may have been derived from flocculent material in the dilated rough endoplasmic reticulum. These observations suggest a structural change in the tumor AAT analogous to that proposed for hereditary AAT deficiency. As a histogenetic marker, the presence of AAT in both epithelial and stromal cells suggests their origin from a common precursor cell.U PMID- 6275724 TI - Rapid detection of viruses in biopsy or autopsy specimens by the pseudoreplica method of electron microscopy. AB - A rapid pseudoreplica method of electron microscopy (EM) was used to detect viruses in brain tissues obtained from 57 patients suspected of having herpes simplex encephalitis (HSE). Herpesvirus particles were demonstrated in 18 of the 20 (90%) specimens from which herpes simplex virus was isolated in tissue culture. The procedure takes less than 2 hours, as compared to the time (greater than or equal to 2 days) usually required for virus isolation or standard thin section electron microscopy. Although the immunofluorescence test has comparable rapidity, it was found to be less sensitive and to yield occasionally false positive reactions. Our findings indicate that, until a brain biopsy can be replaced by less invasive procedures, examination of brain tissue by the pseudoreplica method offers a rapid diagnosis of a herpesvirus encephalitis. Furthermore, the demonstration of viruses in brain, lung and liver specimens obtained by biopsy or at autopsy suggests that this electron-microscopic method would be advantageous for detecting viruses in tissue samples, particularly in the case of viruses which may not be readily demonstrable by conventional methods. PMID- 6275725 TI - A seroepidemiological study of amebiasis in adults in Maracaibo, Venezuela. AB - Eight hundred and forty-eight sera collected from adult patients attending the Central and University Hospitals and from blood donors at the Blood Bank in Maracaibo, Venezuela, were screened for serologic evidence of Entamoeba histolytica infection, employing the indirect hemagglutination test. The seropositivity rates obtained ranged from 4.4-6.5% and most of the positive sera showed low titers of 1:128-1:256. A predominance of positive reactors was observed in the first decades of life with a decline after 44 years of age. Parasitologic examinations made in the Central Hospital population revealed E. histolytica infection in 4% of the patients. PMID- 6275726 TI - Schistosomiasis japonica in Barrio San Antonio, Basey, Samar, in the Philippines. V. The enzyme-linked immunosorbent assay (ELISA) compared with quantitative stool examination and the circumoval precipitin (COP) test. PMID- 6275727 TI - Viruses and bacteria in pediatric diarrhea in Thailand: a study of multiple antibiotic-resistant enteric pathogens. AB - Children with diarrhea admitted to a rehydration ward of a children's hospital in Bangkok were investigated to determine the prevalence of enteric pathogens, the extent of children's previous antibiotic therapy, and the frequency of plasmid mediated antibiotic resistance among infecting bacteria. Rotavirus (36%), enterotoxigenic Escherichia coli (18%), Shigella (9%), Salmonella (6%), Campylobacter jejuni/coli (4%), and Vibrio cholerae (2%) infections were among 105 children with diarrhea. Antibiotics were detected in 29% of urines collected from children on admission. All Shigella, 83% of enterotoxigenic E. coli, and 40% of Salmonella were resistant to more than one antibiotic. Sixty-two percent of 24 antibiotic-resistant enteric pathogens transferred R factors to E. coli K12 by conjugation. Four of four multiresistant E. coli that produced heat-labile and heat-stable toxin transferred resistance and the ability to produce heat-labile toxin. An analysis of plasmids by agarose gel electrophoresis indicated enterotoxigenicity and antibiotic resistance were associated with separate plasmids in transconjugants from these six matings. Antibiotics are used frequently in the treatment of pediatric diarrhea in Bangkok, which has undoubtedly contributed to the high frequency of plasmid-mediated resistance among enteric pathogens. PMID- 6275728 TI - Genetic resistance to lethal flavivirus encephalitis: effect of host age and immune status and route of inoculation on production of interfering Banzi virus in vivo. AB - Phenotypic expression of resistance to Banzi virus infection by C3H/RV mice develops at about 4 weeks of age and occurs when the virus is given at a peripheral inoculation site. In contrast, young or adult congenic C3H/He mice die with an encephalitic illness regardless of the route of inoculation. Data are presented here which suggest that interfering virus (I.V.) production by mice genetically resistant to lethal flavivirus encephalitis may contribute to their survival. The level of I.V. in the brains of weanling C3H/RV mice was lower and I.V. was detected for a shorter period of time than in brains of adult resistant mice. After intracerebral inoculation of virus, to which the C3H/RV mice succumb, I.V. was not detected in the brain at any time after infection. Interfering virus production appeared to originate in cells of the lymphoreticular system with subsequent amplification in the central nervous system. Cyclophosphamide, an immunosuppressive drug which compromises resistance of C3H/RV mice to lethal Banzi virus infection, enhanced production of I.V. in the brains of resistant mice and stimulated I.V. production in the brains of susceptible C3H/He mice. It is hypothesized that I.V. is produced, initially, by a lymphoid cell type which is cyclophosphamide-resistant. PMID- 6275729 TI - [Cytomegalovirus in the salivary glands of the insectivores (author's transl)]. PMID- 6275730 TI - [Placental mechanism of fetal glycemic regulation normally and in asphyxia]. PMID- 6275731 TI - Cutaneous malakoplakia. AB - A case of cutaneous malakoplakia is reported. The patient is a 53-year-old white woman on immunosuppressive therapy 6 years after renal transplantation for end stage nephrosclerosis. She developed yellow-pink papular lesions in her natal cleft which, on histopathologic examination, showed typical features of malakoplakia, namely, a diffuse dermal infiltrate of histiocytes (von Hansemann cells) which contained calcified Michaelis-Gutmann bodies with a concentric ring appearance. Gram-negative bacteria were also identified in some of the histiocytes. The Michaelis-Gutmann bodies stained positively with the PAS diastase, mucicarmine, and Grocott stains and thus resembled fungal spores from which they must be differentiated. Electron microscopy showed that the Michaelis Gutmann bodies developed within phagolysosomes of the histiocytes. Malakoplakia appears to be caused by an acquired defect in the intracellular digestion of phagocytized bacteria. PMID- 6275732 TI - Cutaneous malakoplakia. Report of a case and review of the literature. AB - Cutaneous malakoplakia occurring in the perianal region of a 64-year-old man is reported and the features by light and electron microscopy are described. Review of the literature disclosed that cutaneous malakoplakia is rare, there having been only seven previous cases reported. Current concepts on the pathogenesis of this condition are discussed. PMID- 6275733 TI - Angiofibromas and tuberous sclerosis. PMID- 6275734 TI - Studies on chronic herpetic keratitis--estimation of virus-harboring ganglion cells. AB - The reservoir site of herpes simplex virus (HSV) during the chronic phase of herpetic keratitis was confined to the trigeminal ganglion. Other ocular tissues, the cerebrum and the cerebellum were also examined and not found positive for the isolation of HSV by cocultivation with green monkey kidney cells 28-224 days after infection. The percentage of virus-harboring trigeminal ganglionic cells was estimated by determining the number of dispersed ganglionic cells needed to produce cytopathology of HSV on 50% of Vero cell monolayers. The percentage of infected ganglionic cells were calculated to range from 0.5% to 0.3% during this chronic period. PMID- 6275735 TI - A simple logarithmic amplifier for the photographic quantitation of DNA in gel electrophoresis. PMID- 6275736 TI - 32P-base analysis of DNA. PMID- 6275737 TI - A method for the concentration and for the colorimetric determination of nanomoles of inorganic pyrophosphate. PMID- 6275738 TI - A facile tritium release assay for mammalian L-dihydroorotate dehydrogenase. PMID- 6275739 TI - Enzymatic assays of L-serine in biological material. PMID- 6275740 TI - Differential interferometry in the analytical ultracentrifuge. I. Design, construction, and alignment of differential interferometer. PMID- 6275741 TI - Differential interferometry in the analytical ultracentrifuge. II. General applications. PMID- 6275742 TI - N-chloroacetyl-[125I]iodotyramine: an alkylating agent with high specific activity. PMID- 6275743 TI - A new continuous, monodimensional electrophoretic system for the separation and quantitation of individual glycosaminoglycans. PMID- 6275744 TI - Chemical quantitation of hemoglobin glycosylation: fluorometric detection of formaldehyde released upon periodate oxidation of glycoglobin. PMID- 6275745 TI - The inhibitory effect of isoproterenol and dibutyryl cyclic-AMP on the immediate skin reaction. AB - The concomitant injection of isoproterenol with house dust allergen inhibited the immediate skin allergic reaction in asthmatic patients. Pretreatment with dibutyryl cyclic AMP showed an inhibitory effect on the whealing formation to the allergen. Those results showed the inhibitory effect of isoproterenol on the skin reaction through increasing intracellular cyclic AMP levels. PMID- 6275746 TI - Viral antibody studies of laboratory dogs with diarrheal disease. AB - Viral antibody studies were done on laboratory dogs in an epizootic of gastrointestinal disease. Increased hemagglutination-inhibition antibody titers to a parvovirus (PV) antigenically related to feline panleukopenia virus were found in convalescent serum specimens of 78% (20/26) of the affected dogs and in 83% (5/6) of apparently healthy dogs. With one exception, all dogs tested had significant levels of hemagglutination-inhibition antibody to this PV. Similar increased antibody titers were found to feline panleukopenia virus. Also, neutralizing antibody responses were detected to the canine coronavirus in 24% (6/25) and canine herpesvirus in 45% (10/22) of the affected dogs. However, antibody titers did not increase to canine distemper virus, infectious canine hepatitis virus, canine parainfluenza virus, or minute virus of canines. Subsequent serotesting of the colony provided evidence that additional PV infections occurred in pups from each of 8 litters born 3 to 8 months after the epizootic. These findings indicated the continued presence of the PV for more than 1 year in the infected colony. Of 19 laboratory personnel who worked with the affected dogs, none, including 4 with a concurrent diarrheal disease, developed or had antibodies to the PV or canine coronavirus. PMID- 6275747 TI - Canine infectious tracheobronchitis: effects of an intranasal live canine parainfluenza-Bordetella bronchiseptica vaccine on viral shedding and clinical tracheobronchitis (kennel cough). AB - A modified-live intranasal (IN) canine parainfluenza (CPI)-virus Bordetella bronchiseptica vaccine was evaluated in dogs for efficacy against laboratory induced canine infectious tracheobronchitis. The comparative efficacies of IN and parenteral administrations of the CPI virus fraction were also evaluated. The frequency and duration of clinical tracheobronchitis, blood serum agglutination titer, humoral antibody response, and duration of CPI virus and B bronchiseptica shedding were measured. Group A dogs were vaccinated subcutaneously or IM with an experimental CPI vaccine and challenge exposed with CPI virus. Group B dogs were vaccinated IN with avirulent CPI virus-B bronchiseptica live antigens and challenge exposed with virulent CPI virus and virulent B bronchiseptica. The IN vaccination (group B) significantly reduced (P less than or equal to 0.001) the occurrence of clinical tracheobronchitis by 96%. The combined challenge exposure of virulent CPI and virulent B bronchiseptica produced a synergistic enhancement of the clinical signs of kennel cough. The percentage of days after challenge exposure that virus shedding was detected for controls equaled 70% as compared with 50% and only 1% for parenterally and IN vaccinated dogs, respectively. Isolation of virulent B bronchiseptica microorganisms was reduced 89% in dogs vaccinated IN compared to controls. The geometric mean humoral antibody titers to CPI virus after 2 parenteral vaccinations and 1 IN vaccination were 1:43 and 1:34, respectively. PMID- 6275748 TI - Resistance of porcine preimplantation embryos to pseudorabies virus. PMID- 6275749 TI - Intrauterine transmission of ovine progressive pneumonia virus. AB - Ovine fetuses, newborn lambs, and ovine colostrum were examined for ovine progressive pneumonia virus. The lambs and colostrum were also examined for specific antibody. Virus was isolated from 1 fetus, from 2 newborn lambs, and from most samples of colostrum. The fetus was about 100 days old and was carried by a seronegative ewe in contact with seropositive sheep. Both newborn lambs were carried by seropositive ewes. One lamb was dead at birth; the other lamb was normal and had not nursed. Antibody specific for the virus was present in the colostrum of 12 of 14 seropositive ewes and in the serum of 8 of 11 lambs that had nursed seropositive ewes, but not in the serum of lambs that had not nursed. PMID- 6275750 TI - Further studies of naturally occurring latent bovine herpesvirus infection. AB - Attempts were made to enhance the isolation rate of latent bovine herpesviruses from trigeminal ganglia by using a fibroblastic fetal bovine kidney cell line, in addition to Madin-Darby bovine kidney cells, and by superinfecting with a temperature-sensitive helper strain of bovine herpesvirus-1 (BHV-1). Four isolates of latent BHV-1 were obtained from 44 pairs of trigeminal ganglia--thus reflecting no increase in isolation rate over that previously observed. Two isolates of a latent bovine herpesvirus-3 strain DN599 were also obtained. Analysis of plaque morphology of BHV-1 isolates obtained in this and a previous study indicated that several biologically heterogeneous strains were capable of establishing latent infections. PMID- 6275751 TI - Egg transmission of avian adenovirus-associated virus and CELO virus during experimental infections. PMID- 6275752 TI - Bovine leukemia virus infection in a large Holstein herd: cohort analysis of the prevalence of antibody-positive cows. PMID- 6275753 TI - Bovine leukemia virus infection in a large Holstein herd: prospective comparison of production and reproductive performance in antibody-negative and antibody positive cows. AB - Serum samples from lactating cows in a purebred Holstein herd were tested annually (from 1977 to 1980) for antibodies to bovine leukemia virus (BLV), using the agargel immunodiffusion test. Production and reproductive variables were obtained from Dairy Herd Improvement Association records. All milk and fat production values were converted to 3.5% fat-corrected milk (FCM). Variables examined included: FCM, 305-day actual; FCM, 305-day mature-equivalent; FCM, total lactation; FCM per day, 305-day actual; total days milked during lactation; days nonlactating; age at calving; calving interval; days open, and number of times bred. Lactations were stratified from 1 to greater than 5 for comparison of variables. A matched case-control analysis was performed to assess the risk of clinical mastitis in BLV-infected cows. The retention of BLV antibody-negative and antibody-positive cows in the herd was compared. There were no significant trends in the means of production and reproductive variables between BLV antibody negative and antibody-positive cows. The relative risk of clinical mastitis in BLV antibody-positive cows was 1.3, which was not significant (P greater than 0.05). Survivorship analysis over 3 years demonstrated no significant difference in the retention of BLV antibody-negative and antibody-positive cows in the herd. The BLV-infected cows did not have lower milk production, poorer reproductive efficiency, increased prevalence of mastitis, or lesser longevity in the herd than did noninfected cows. PMID- 6275754 TI - Effect of colostral antibody on bovine leukemia virus infection of neonatal calves. AB - Pairs of newborn calves were exposed to bovine leukemia virus (BLV) when they were given their 1st colostrum feeding. Calves that were given 10(6) BLV-infected lymphocytes in colostrum free of BLV-specific antibody became infected. Calves that were fed 10(7), 10(8), or 10(9) infected lymphocytes in colostrum that contained BLV-specific antibody did not become infected. One of 2 calves inoculated intradermally with 250,000 infected lymphocytes was protected by colostral antibody, but the other was not. Colostral antibody titers in the unprotected calf decreased normally until the calf was 4 months old and then increased markedly; this pattern indicates that the presence of colostral antibody may have prolonged the latent period of the BLV infection. PMID- 6275755 TI - Stability of viability and immunizing potency of lyophilized, modified equine arteritis live-virus vaccine. AB - The Bucyrus strain of equine arteritis virus, previously modified to avirulence and vaccinal virus by 131 serial passages in primary cell cultures of horse kidney followed by 111 passages in primary cell cultures of rabbit kidney, was further passaged in cultures of the E. Derm (NBL-6) cell line, a continuous diploid cell line. Pools of the 16th and 25th passages of the virus in this last equine dermal cell line were lyophilized and stored in lots at 37 C, 23 to 28 C, 4C, and -20 C. The viability of the vaccinal virus deteriorated rapidly during storage at 37 C and at 23 to 28 C, but was relatively stable at 4 C and at -20 C for at least 1 year. The vaccine stored at 4 C for 9 months protected 2 horses, subsequently inoculated with avirulent virus, from arteritis, whereas the 2 nonvaccinated control horses inoculated simultaneously developed severe signs of the disease and died of acute arteritis. PMID- 6275757 TI - Displacement of protein-bound thiopental by sodium methiodal may contribute to anesthetic complications during canine myelography. PMID- 6275756 TI - Ovine progressive pneumonia: pathologic and virologic studies on the naturally occurring disease. AB - Pathologic and virologic studies were conducted on 13 mature ewes with serum precipitin antibodies to progressive pneumonia virus (PPV). Pulmonary lesions of ovine progressive pneumonia were found in 4 sheep, a meningoencephalitis resembling visna in 1 sheep, chronic proliferative carpal arthritis in 2, and massive lymphoid proliferation in the mammary gland in 3. Virus producing cytopathic effect typical of PPV was isolated from the lungs, mediastinal lymph node, spleen, and choroid plexus of 4 sheep and from the carpal synovium of 2 sheep with chronic carpal arthritis. Three viral isolates selected for further study were antigenically related to visna virus by immunofluorescence and immunodiffusion, but these 3 isolates were not neutralized by antisera to reference strains of visna virus. Seemingly, infection of sheep by ovine retroviruses is common in the United States, and these viruses are capable of causing disease in more than 1 organ system. PMID- 6275758 TI - Effects of storage on concentration of hydrocortisone (cortisol) in canine serum and plasma. AB - A specific radioimmunoassay was used to measure concentrations of hydrocortisone (cortisol) in the serum and plasma of 4 dogs. Differences (P greater than 0.05) in concentrations of cortisol were not found between serum and plasma (from EDTA treated and heparinized blood samples). Differences (P greater than 0.05) in serum or plasma concentrations of cortisol were not found between samples stored at 4 C for various times (10 minutes, 10 hours, 40 hours) after collection, but before removal of RBC. In a study designed to determine the stability of cortisol in serum samples stored at room temperature, degradation was dependent on the initial serum concentrations of cortisol. Decreases (P greater than 0.05) did not occur in concentrations of cortisol in serum samples stored up to 15 days when initial concentrations of cortisol were less than 15 ng/ml. However, when initial concentrations of cortisol were approximately 55 ng/ml and 80 ng/ml, significant (P greater than 0.05) degradation occurred after 9 and 5 days of storage, respectively. Results of this investigation indicate that either serum or plasma of dogs is suitable for radioimmunoassay of cortisol and that samples (with and without added coagulants) incubated at 4 C may be left uncentrifuged for up to 40 hours without cortisol degradation. However, prolonged storage of serum at room temperature is detrimental, particularly for samples having large concentrations of cortisol. PMID- 6275760 TI - Reference material for the quantitation of alpha quartz in respirable dust by x ray diffraction. PMID- 6275759 TI - Plasmid DNA content of multiresistant Staphylococcus aureus strains. AB - Multiresistant S. aureus strains (also called methicillin-resistant in clinical laboratories) share many common properties. In addition to the commonly used epidemiological markers (pattern of resistance to antibiotics and metal ions, serotype and lysotype), we considered using the molecular and genetical properties of the plasmid DNA harboured by these strains. In this paper we report an epidemiological study of two groups of S. aureus strains sampled since 1975 and carrying new resistance characters which appeared with increasing frequency in France. The Tm-SgR strains (5 strains) carry, in addition to 13 resistance characters commonly observed, two new characters: the resistance to gramin components A and B (Sg). The two new characters were shown to be encoded by a single plasmid. Amongst the 5 strains studied, two harbour a similar plasmid DNA content while the three others were clearly distinct. Because of the variability of the plasmid DNA content observed in these strains, the similarity of this content observed between two of them suggest a close phylogenic relationship. These two strains are probably derivatives from a single clone. The spread of the two new characters amongst the distinct strains might be related to the spread of ancestrally related Tm-Sg plasmids. The Tob-GenR strains (16 strains) carry, in addition to 12 wide-spread resistance markers, the newly observed resistance to many aminoglycosides, mainly the resistance to tobramycin (Tob) and gentamicin (Gen). They all harbour a 22-kb plasmid DNA generating similar restriction cleavage patterns and conferring the resistance to penicillin G, arsenate, arsenite and cadmium. Some of them harbour one or two extra small plasmids (3.4 and 2.3 kb); the presence of these two plasmids was not associated with a modification of the resistance pattern of the strains. Most of the resistance characters--including the new ones--are probably encoded by chromosomal genes. In this epidemiological situation, the spread of the new aminoglycoside resistance characters carried by the 16 strains having the same phenotype and harbouring the same 22-kb plasmid is attributed to the spread of derivatives from a single clone. PMID- 6275761 TI - [Signal symptoms in tumours of the petrous bone (author's transl)]. AB - Supra-labyrinthic tumours usually produce lesions of the Vth and VIth cranial nerves, whereas infra-labyrinthic tumours affect the sensory-motor nerves. Both types of tumour may reveal themselves by cochleo-vestibular disorders and/or middle ear symptoms. The author has treated a series of patients with such tumours and describes the most frequent types of intrapetrosal growth: cholesteatoma (12 cases), glomic tumours (11 cases), facial nerve tumours (3 cases), metastases (2 cases). Meningiomas can also be encountered, as well as rarer tumours (4 cases), such as embryonic sarcoma, chordoma, chondroma and chondrosarcoma, cavernous angioma, eosinophilic granuloma, solitary plasmocytoma and fibrous dysplasia of the petrous bone. Some signal symptoms (sudden deafness, mucous otitis media, paralysis of vocal cords) can be particularly misleading. PMID- 6275762 TI - [Diagnosis of glomus jugulare tumours (author's transl)]. AB - The present study is based on 16 personal cases and on an analysis of the principal series published since 1968. Clinical investigations aim at detecting lesions of the sensory-motor and XIIth cranial nerves, deafness which indicates invasion of the labyrinth, lesions of central auditory and vestibular pathways and possible associated conditions. Radio-otology is fundamental in revealing alterations of the jugular foramen and above all perhaps, changes in the tympanic bone and in the 3rd segment of the fallopian canal. The most significant sign of invasion is infra-labyrinthic hollowing out on tomographic sections in lateral projection and Guillen's incidence. Angiography and jugulography supply information on the size of the tumour and on its feeding vessels which usually arise from the external carotid artery but also, in advanced forms, from the internal carotid and vertebral arteries. Computerized tomography for the time being has one single major applications : detection of lesions extending to the posterior and middle fossae. Taken together, these investigations should help to classify tumours of the glomus jugulare into : exclusively tympanic, tympano mastoidal, tumours extending posteriorly and to the petrous bone, and tumours extending intracranially. Such a classification is of paramount importance in deciding on the best surgical approach. PMID- 6275763 TI - [Round table. Microsurgery and problems of tumors of the base of the skull]. PMID- 6275764 TI - [Radiotherapy in cases of tumours glomus jugulare (author's transl)]. AB - Eight cases of glomus jugulare tumours were irradiated with megavoltage from 1964 through 1978. All had extensive bone destruction of the base of skull 5 000 to 6 000 rads in 7 of the cases. Regression of functional signs was constant whereas regression of palsies was rare and recalcification of bone lesions exceptional. All patients are living with apparent control of disease for periods of 16 to c years. Accurate technique is indispensible to prevent radiation myelitis. For the extensive jugulare tumours results of radiotherapy compare very favorably with those of extensive surgery. PMID- 6275765 TI - [Atopic dermatitis and pox-virus superinfections (author's transl)]. AB - Subjects afflicted with atopic dermatitis are particularly predisposed to such viral superinfections as verruca, herpes (eczema herpeticum) and vaccinia (eczema vaccinatum). Knowledge of superinfection by the molluscum contagiosum virus is more recent and the authors describe one case. The orf virus (responsible for ovine ecthyma contagiosum), however, has never been described as responsible for the profuse superinfection of an atopic eczema. The authors observed one case of this in a child, atypical as result of his age (16 months), the cervical localization and the aspect, which was a telangiectasic granuloma type of the initial element, and finally of the satellization observed one week later. This child suffered from a generalized atopic eczema. Propagation of the orf lesions could have been favored by scratching, local corticoid treatment and by the deficit in cellular immunity observed in patient with atopic dermatitis. Indeed, viral superinfections, which are unusual by their profusion or their chronic nature, are often observed in immunodepressed subjects. PMID- 6275766 TI - [Cutaneous blastomycosis and its treatment. Apropos of a new Moroccan case]. PMID- 6275767 TI - [Parotid tumours. Only 31% of mixed tumours. In one hundred and seventy-five parotidectomies (author's transl)]. PMID- 6275768 TI - Vasoactive intestinal peptide (VIP) secreting tumour of the pancreas. AB - A 51-year-old woman with a history of several years' watery diarrhoea caused by a vasoactive intestinal peptide (VIP) secreting tumour of the pancreas (VIPoma) is presented. Measured stool outputs ranged from 1000 to 2600 ml/24 h. S-K 2.5 mmol/I. The pancreatic VIPoma was localised by abdominal angiography and removed. The plasma concentration of vasoactive intestinal peptide was 80 pmol/l before and 5 pmol/I after the operation (normal under 30 pmol/I). The patient has been symptom-free for 15 months since surgery. PMID- 6275769 TI - Diethyldithiocarbamate in treatment of acute cadmium poisoning. AB - An insoluble complex was synthesized by the reaction of sodium diethyldithiocarbamate trihydrate (DDTC) with CdCl2 X 2.5 H2O (Cd). Elemental analyses of the product yielded a percent composition of each element which was consistent with the postulated structure of two mole equivalents of (DDTC)- and one mole equivalent of Cd++. The DDTC formed a complex with 11 other metal ions tested, but not with Ca++ or Mg++. The complex with Cd was highly insoluble in water, 0.1 N HCl, 0.1 N NaOH, human serum, and carbon tetrachloride, but it was soluble in pure dimethyl sulfoxide. The DDTC protected mice from a greater than LD100 dose of Cd. This protective effect was more pronounced when DDTC treatment was delayed at least 30 min after i.p. administration of Cd. More than 98 percent of mice treated with 500 mg per kg of DDTC 30 min to five hrs after Cd administration survived, and 50 percent survived when treatment wad delayed for eight hrs. Administration of DDTC prior to or immediately after Cd gave less protection. The lowest (DDTC)-/Cd++ molar dose ratio which resulted in 100 percent survival following the otherwise lethal dose of Cd was 7.6, which was less than four times the theoretical stoichiometric molar ratio for formation of the complex. The LD50 of the complex in mice wa approximately 650 mg per kg of the i.p. route, indicative of a low degree of dissociation in vivo. PMID- 6275771 TI - The origin of spontaneous electrical activity at the end-plate zone. AB - Two types of spontaneous electrical activity are present at the end-plate zone: low-voltage negative potentials that correspond to miniature end-plate potentials, and larger voltage negative-positive potentials. The electrogenic origin of the latter has been uncertain. The origin of these larger potentials was investigated in the rat phrenic nerve diaphragm preparation and in human gastrocnemius muscle just prior to intubation during administration of preoperative anesthesia. In the hemidiaphragm the larger voltage negative positive potentials were rarely triggered by intracellular or tungsten microelectrodes. The negative-positive potentials, however, were clearly triggered by contact of the concentric needle electrode with muscle hemidiaphragm at the end-plate region. The potentials were abolished by curare. Likewise, the equivalent potentials observed at the human gastrocnemius end-plate zone were blocked by neuromuscular blocking agents. Therefore, these positive-negative discharges represent postsynaptic muscle fiber action potentials and not nerve fiber activity. They were probably presynaptically activated by mechanical irritation of the motor axon terminal and preterminal branches. PMID- 6275770 TI - Dependence of the Achilles tendon reflex on the excitability of spinal reflex pathways. AB - Muscle afferent activity from the triceps surae was recorded during experimentally induced alterations in amplitude of the Achilles tendon jerk. No changes in the neural afferent response to tendon percussion or in the background level of neural activity occurred when the reflex response was altered by discomfort, distraction, changes in attention, or changes in the rate of tendon percussion. Reinforcement of the Achilles tendon jerk by forceful contraction of the forearm muscles did not alter the relationship between intensity of the tendon tap and amplitude of the evoked neural afferent volley. Nevertheless, such maneuvers lowered the reflex threshold and raised reflex sensitivity so that a smaller afferent volley was required to produce a tendon jerk, and an increase in the afferent volley produced a disproportionately greater increase in reflex electromyographic activity than would have occurred at rest. Reinforcement maneuvers potentiated the H-reflex but did not alter the electrically induced afferent volley or the background level of neural activity. It is concluded that these changes in reflex responsiveness occurred through intrinsic spinal mechanisms independent of the fusimotor system. PMID- 6275772 TI - [Structure of the tricyclic triaminotricarboxylic acid (Y) from the antibiotic actinoidin]. AB - The products of triaminotricarboxylic acid (Y) transformation in alkaline and reduction (HI/P) hydrolysis and the PMR spectra of the Y-Phe dipeptide were studied and the nature of the 3 amino acid substitutes and their position in the aromatic nuclei were determined. With regard to the data on the structure of the aromatic skeleton published earlier the structure of the Y amino acid as 3-(2 chlor-4-seryl-phenoxy)-5-(4-seryl-phenoxy)-p-hydroxyphenylglycine was suggested. The presence of 2 alcoholic groups of the phenylserine fragments of the Y amino acid in the actinoidine aglycone was shown with additional acetylation (Ac2O/Py) of N-acetylmethoxyaglycone. It was found with O-acetylation of N acetylmethoxyactinosaminyl aglycone that the actinosamine amino sugar in the actinoidine molecule was bound with one of the above groups. PMID- 6275773 TI - [Carbohydrate linkage with the peptide portion of the ristomycin A molecule]. AB - The mannose monosaccharide residue in ristomycin A is bound via C-3 phenolic oxygen of the 1,2,3,5-substituted ring with actinoidinic amino acid. The tetrassacharide branch is attached to the only phenolic oxygen of triaminotricarboxylic acid. The ristosaminyl residue is linked via one of the two aliphatic hydroxyl groups of the same amino acid. By the location of neutral carbohydrates risotmycin A does not differ from ristocetin A. PMID- 6275774 TI - [Achievements in the field of obtaining and studying new semisynthetic cephalosporin antibiotics. Research in the field of directed cephalosporin synthesis]. PMID- 6275775 TI - [Migration of the ampicillin transposon in enteropathogenic Escherichia]. AB - Migration of Tn 1 from plasmid RP4 into the chromosome of enteropathogenic Escherichia (EPE) of serogroups O124 and O111 was studied. E. coli K 12 LC 411 (RP4) was used as the donor. It was shown that the transposition rate markedly differed depending on the period of ;the cell isolation from Tn 1 in the chromosome, i. e. during conjugation or after several subcultures of the transconjugants from the autonomic plasmid onto the selective media. During the conjugation process the migration rate of Tn 1 was equal to 0.5 and 0.8 per cell acquiring the plasmid. The transposition rate in the EPE carrying the R factor for a long period of time was 2.4 . 10(-2) and 1.4 . 10(-2) respectively for each serogroup. The above differences in the migration rate of Tn 1 were not concerned with changes in the environment. PMID- 6275776 TI - Comparative multiple-dose pharmacokinetics of cefotaxime, moxalactam, and ceftazidime. AB - The pharmacokinetics of cefotaxime, moxalactam, and ceftazidime were investigated in six human volunteers who received in a crossover fashion doses of 0.5, 1.0, and 2.0 g of each drug by a 5-min infusion. Doses of 1.0 g were repeated after the administration of probenecid. Serum and urine concentrations were assayed with an agar diffusion method. Serum concentrations of moxalactam exceeded those of ceftazidime at all times and were distinctly higher than those of cefotaxime. The normalized area under the concentration time curve (defined as the ratio of the area under the curve per dose) reflects this relationship: compared with cefotaxime the normalized area under the curve of moxalactam was 3 to 4 times higher, and that of ceftazidime was 2 to 3 times higher. By intra-individual comparisons, the area under the curve of moxalactam was 44% larger than that of ceftazidime. With increasing doses, cefotaxime exhibited a nonlinear increase of the area under the curve. The half-lives of moxalactam, ceftazidime, and cefotaxime were 2.34, 1.95, and 1.16 h, respectively. The volume of distribution averaged 0.20 +/- 0.03, 0.23 +/- 0.02, and 0.25 +/- 0.04 liters per kg, and the mean total body clearance was 84, 131, and 328 ml/min for moxalactam, ceftazidime, and cefotaxime, respectively. The 24-h urinary recovery was highest for moxalactam (75 +/- 4%) followed by ceftazidime (68 +/- 11%) and cefotaxime (53 +/- 6%). The influence of probenecid on serum concentrations, half-life, area under the curve, and clearance was most apparent with cefotaxime, whereas the pharmacokinetics of moxalactam and ceftazidime were only slightly affected. After the 0.5- and 2.0- g doses of cefotaxime, desacetyl-cefotaxime activity (determined by high-pressure liquid chromatography) reached a peak of 2.7 and 9.9 mug/ml and declined with a half-life of 1.9 and 1.4 h. The ratio of the R(-) and S(-) epimers of moxalactam, which could be differentiated by high-pressure liquid chromatography, fell rapidly from 0.81 at 0.17 h to 0.5 at 5 h, indicating the presence of twice as much of the microbiologically less active S(-) epimer. From a pharmacokinetic standpoint it appears reasonable to conclude that moxalactam and possibly ceftazidime could be administered twice daily and that cefotaxime could be administered three or even four times daily. PMID- 6275777 TI - Quantitation of herpes simplex virus type 1 shed in preocular tear film of rabbits treated with acyclovir. AB - The quantity and duration of herpes simplex virus type 1 shedding in the preocular tear film of rabbits were measured before, during, and after administration of acyclovir topically, intravenously, and by these routes. Topical administration reduced shedding significantly, Intravenous administration was without effect and in the combination regimen added nothing to the effectiveness of local application. The effects of acyclovir were temporary because there were significant increase in preocular tear film virus shedding after cessation of treatment. PMID- 6275778 TI - Selective suppression of alimentary tract microbial flora as prophylaxis during granulocytopenia. AB - Oral nonabsorbable antibiotics have been used to suppress the rectal flora in granulocytopenic patients. Problems with these therapies, i.e., compliance, acquisition of undesirable flora, and cost, motivated the search for an alternative therapy which would increase compliance and effectively reduce the Enterobacteriaceae without creating a microbiol vacuum. Trimethoprim sulfamethoxazole was found to be easily taken, to suppress the Enterobacteriaceae, and to maintain the anaerobic rectal flora for biological stability of the rectal ecosystem. However, concurrent use of parenteral antibiotics profoundly influenced rectal flora and temporarily destroyed the colonization resistance afforded by the anaerobes. PMID- 6275779 TI - Pharmacokinetics of ceftriaxone in humans. AB - Pharmacokinetics of the investigational cephalosporin ceftriaxone were studied after 30-min intravenous infusions of three ascending single doses of 0.5, 1, and 2 g crossed over in 12 normal subjects. Serially collected plasma and urine samples were analyzed for ceftriaxone by high-performance liquid chromatography. Plasma concentration-time profiles were characterized by a linear two-compartment open model with the following respective mean (+/- standard deviation) parameters at 0.5-, 1-, and 2-g dose levels: elimination half-life, 6.5 +/- 0.7, 6.2 +/- 0.8, and 5.9 +/- 0.7 h; apparent volume of distribution, 8.5 +/- 1.1, 9.0 +/- 1.1, and 10.1 +/- 1.0 liters; and plasma clearance, 929 +/- 150, 1,007 +/- 130, and 1,190 +/- 150 ml/h. The respective renal excretion parameters were as follows: renal clearance, 373 +/- 60, 399 +/- 50, and 533 +/- 128 ml/h; and percentage of dose excreted unchanged in the 48-h urine samples, 41 +/- 8, 39 +/- 5, and 43 +/- 10. The 6-h elimination half-life of ceftriaxone was 2- to 10-fold longer than those reported for marketed and other known investigational cephalosporins. The small dose-related increases in the apparent volume of distribution and clearance parameters can be explainhe 48-h urine samples, 41 +/- 8, 39 +/- 5, and 43 +/- 10. The 6-h elimination half-life of ceftriaxone was 2- to 10-fold longer than those reported for marketed and other known investigational cephalosporins. The small dose-related increases in the apparent volume of distribution and clearance parameters can be explainhe 48-h urine samples, 41 +/- 8, 39 +/- 5, and 43 +/- 10. The 6-h elimination half-life of ceftriaxone was 2- to 10-fold longer than those reported for marketed and other known investigational cephalosporins. The small dose-related increases in the apparent volume of distribution and clearance parameters can be explained by the concentration-dependent plasma protein binding of ceftriaxone in humans. The impact of the small dose-dependent changes in the pharmacokinetics of ceftriaxone is anticipated to be of negligible clinical significance. PMID- 6275781 TI - Comparative in vitro activity of cefmenoxime, cefotaxime, cefuroxime, cefoxitin, and penicillin against Neisseria gonorrhoeae. AB - The in vitro activity of a new beta-lactam antibiotic, cefmenoxime, was compared with those of cefotaxime, cofuroxime, cefoxitin, and penicillin against 72 beta lactamase-negative and 26 beta-lactamase-positive Neisseria gonorrhoeae. Cefmenoxime was as active as cefotaxime and more active than the other three antimicrobial agents. It inhibited all isolates, regardless of beta-lactamase activity, at a concentration of less than or equal to 0.015 microgram/ml. PMID- 6275780 TI - Ketoconazole, amphotericin B, and amphotericin B methyl ester: comparative in vitro and in vivo toxicological effects on neutrophil function. AB - We investigated a number of parameters for host defense after the in vitro addition of the antifungal agents ketoconazole, amphotericin B (AMB), and amphotericin B methyl ester (AME). Similar assays were repeated before and after patients received the former two drugs. Viability by trypan blue exclusion, adherence by nylon wool columns, chemotaxis by the under-agarose technique, phagocytosis and killing by chemiluminescence, colony counts, and acridine orange direct visualization were assayed. In striking contrast to AMB and AME, ketoconazole demonstrated no significant effect on neutrophils. Adherence in the presence of therapeutic plasma levels of AMB and AME was decreased (P less than or equal to 0.005) at low drug concentrations, whereas at higher concentrations, adherence was increased (P less than 0.001). The chemotactic responses of cells incubated with AMB and AME demonstrated marked suppression. Phagocytic capacity and killing were decreased (P less than or equal to 0.005) with AMB as compared with control assays and assays performed in the presence of ketoconazole and AME. However, no difference were observed between two patients who received AMB and two other treated with ketoconazole. PMID- 6275782 TI - Comparative in vitro activity of ceftizoxime, cefoperazone, and cefoxitin against anaerobic bacteria. AB - Against 482 obligate anaerobes studied by the agar dilution technique, ceftizoxime was significantly more active than both cefoxitin and cefoperazone (P less than 0.001); the latter two agents were comparable in activity. The enhanced activity of ceftizoxime, as compared with the activity of cefoxitin, was against both gram-positive and gram-negative anaerobes (especially Lactobacillus and bacteroides spp). Cefoperazone, however, was more active than cefoxitin against gram-positive anaerobes (particularly Lactobacillus spp.) but was less active than cefoxitin against gram-negative anaerobes (particularly Bacteroides fragilis and Veillonella spp.). PMID- 6275783 TI - Susceptibility of zygomycetes to amphotericin B, miconazole, and ketoconazole. AB - Susceptibility to amphotericin B, miconazole, and ketoconazole was determined for 25 clinical isolates of zygomycetes in yeast nitrogen base broth and human serum. In yeast nitrogen base broth, 7 of 25 isolates were susceptible to an imidazole or amphotericin B. In serum, all were resistant to the imidazoles and inhibited by amphotericin B, the current drug of choice for infections caused by zygomycetes. PMID- 6275784 TI - Cefmenoxime (SCE-1365), a new cephalosporin: in vitro activity, comparison with other antimicrobial agents, beta-lactamase stability, and disk diffusion testing with tentative interpretive criteria. AB - The in vitro activity of cefmenoxime (SCE-1365) was evaluated in a multiphased collaborative investigation. Over 7,500 consecutive clinical isolates were tested in five laboratories, and greater than 90% of the following organisms were inhibited by cefmenoxime at the following concentrations: Enterobacteriaceae and non-enterococcal streptococci, /=22 mm, susceptible; 1.0). In short, amino acid transport in neonatal rat hepatocytes is increased as a result of an existing low-K(m) component for the Na(+)-dependent alpha-aminoisobutyrate uptake, which endows the hepatocytes with a high capability for concentrating amino acids at low ambient values. The concomitant enhancement of K(+) transport reflects changes in the electrochemical gradient for Na(+) across the hepatocellular membrane and, along with this, presumably alterations in the membrane potential; the latter might be the driving force for the enhanced alpha aminoisobutyrate transport in the alanine-preferring system during postnatal age. PMID- 6275851 TI - Effects of micromolar concentrations of free calcium ions on the reduction of heart mitochondrial NAD(P) by 2-oxoglutarate. AB - 1. The reduction of mitochondrial NAD(P) by 2-oxoglutarate was monitored as a measure of 2-oxoglutarate dehydrogenase activity in its intramitochondrial locale. In the absence of ADP, steady-state reduction of NAD(P) by 0.5 mM-2 oxoglutarate in the presence of 0.5 mM-L-malate was markedly increased by extramitochondrial Ca2+, with 50% activation at pCa 6.58, when the Na+ concentration was 10 mM, the Pi concentration ws 5 mM and the added Mg2+ concentration was 1 mM. Omission of Pi resulted in 50% activation at pCa 6.77; omission of Mg2+ resulted in 50% activation at pCA greater than or equal to 7.3. 2. The activation of 2-oxoglutarate dehydrogenase could be reversed on addition of an excess of EGTA. The rate of inactivation was dependent on the concentration of Na+, with K0.5 2.5 mM, which is consistent with the rate of withdrawal of Ca2+ from the mitochondria being the limiting factor. 3. The steady-state reduction of cytochrome c by 2-oxoglutarate (0.5 mM) also showed a marked dependence on pCa in the absence of ADP; in the presence of an excess of ADP, no such effect of Ca2+ was detectable. 4. Mitochondria from the hearts of senescent rats showed an undiminished rate of dehydrogenase activation by Ca2+ but a rate of inactivation by excess EGTA that was diminished by 40%. Direct studies of Ca2+ egress with Arsenazo III confirmed a decrement in rate with old age. 5. Studies of 2 oxoglutarate dehydrogenase activity as a function of the mitochondrial context of Ca2+, as measured by atomic-absorption spectrophotometry, showed half-maximal activation at a mitochondrial content of 1.0 nmol of Ca2+/mg of protein, and saturation at 3 nmol/mg. 6. These findings support the model advanced by Denton, Richards & Chin [(1978) Biochem. J. 176, 899-906], of a control of the tricarboxylate cycle by intramitochondrial Ca2+, and demonstrate the range of mitochondrial Ca2+ content over which this may occur. In addition, they raise the possibility of a disturbance of this control mechanism in old age. PMID- 6275852 TI - Renal glycerol metabolism and the distribution of glycerol kinase in rabbit nephron. AB - Glycerol and dihydroxyacetone are metabolized by rabbit kidney-cortex tubules, isolated by collagenase treatment. Half-maximal concentrations of both substrates were determined with regard to uptake rates and product formations. Maximal uptake rates were 643 and 329 mumol/h per g of protein for dihydroxyacetone and glycerol respectively. Glucose and lactate were found as major metabolic products. Glycerol kinase, the enzyme catalysing the first step in renal glycerol and dihydroxyacetone metabolism, was measured radiochemically as described by Newsholme, Robinson & Taylor [(1967) Biochim, Biophys. Acta 132, 338-346] and adapted for studies of the localization of this enzyme along the different structures of rabbit nephron. The results show that glycerol kinase is located exclusively in the proximal segments, i.e. the proximal convoluted tubules and the pars recta, but is negligible in the other structures studied. The activities were close to the maximal dihydroxyacetone uptake rates measured in tubule suspensions. PMID- 6275853 TI - Rates of pinocytic capture of simple proteins by rat yolk sacs incubated in vitro. AB - The rates of pinocytic uptake of a number of small 125I-labelled simple proteins (insulin, ribonuclease A and lysozyme) by rat yolk sacs incubated in vitro were determined both before and after treating these proteins with reagents that are known to increase the rate of capture of 125I-labelled bovine serum albumin. Uptake of the untreated forms of all three proteins was extremely rapid, indicating that adsorptive pinocytosis is the principal mechanism by which yolk sac cells capture these simple proteins, but these rates show no simple correlation with molecular charge. In contrast with albumin, the rates of uptake of treated proteins were either unchanged or lower than that of the corresponding untreated protein preparations; polymeric forms of 125I-labelled lysozyme larger than dimers were ingested at rates significantly lower than that of the monomer. PMID- 6275854 TI - An investigation of sites that bind human somatotropin (growth hormone) in the liver of the pregnant rabbit. AB - The binding of 125I-labelled human somatotropin (growth hormone) to a crude membrane preparation from the liver of pregnant rabbit, and to receptors solubilized from this fraction by Triton X-100, was dependent on time, temperature and receptor concentration. At 4 degrees C a steady state was reached after 20 h, and maximum specific binding (as a percentage of total tracer added) was approx. 50% for both membrane-bound and solubilized receptors. Solubilization did not significantly affect the binding properties of the receptor at low concentrations of Triton X-100 (less than 0.05%, v/v, in the assay tube). However, at higher concentrations (approx. 0.1%, v/v), the detergent lowered the ability of some hormones, for example ovine prolactin, to displace 125I-labelled human somatotropin, but did not affect other hormones such as bovine somatotropin. Some somatogenic hormones, such as bovine somatotropin, and some lactogenic hormones, such as ovine prolactin, displaced 125I-labelled human somatotropin from membrane-bound and solubilized receptor preparations. Furthermore, 85% of 125I-labelled bovine somatotropin was displaced from membrane bound receptors by ovine prolactin, and 125I-labelled ovine prolactin was almost completely displaced by bovine somatotropin. Scatchard analysis of the binding data for human somatotropin suggested a single class of binding sites in the membrane-bound receptor preparation, with an affinity (Ka) of 1.9 X 10(9) M-1 and a capacity of 1726 fmol/mg of protein; these values were slightly increased by solubilization (Ka = 3.2 X 10(9) M-1, capacity = 2103 fmol/mg of protein). Scatchard analysis of binding to membrane-bound receptors also indicated a single class of high-affinity binding sites for bovine somatotropin (Ka = 4.8 X 10(9) M 1, capacity = 769 fmol/mg) and for ovine prolactin (Ka = 6.1 X 10(9) M-1, capacity = 187 fmol/mg). PMID- 6275855 TI - Maximum activities and properties of glucose 6-phosphatase in muscles from vertebrates and invertebrates. AB - 1. The maximum catalytic activities of glucose 6-phosphatase were measured in a large number of muscles from vertebrates and invertebrates. The activities range from less than 0.1 to 8.0 mumol/min per g fresh wt. at 30 degrees C: the highest activity, observed in the flight muscle of the wasp (Vespa vulgaris), is similar to that in rat liver. The hydrolytic activity was shown to be specific towards glucose 6-phosphate. 2. The pH optimum was 6.8 and the Km was approx. 0.6 mM (flight muscle of a moth). 3. Almost all of the glucose 6-phosphatase activity from extracts of the flight muscle of a moth and the pectoral muscle of a pigeon were recovered in the cytosolic fraction (i.e. 150,000 g supernatant). 4. During development of the locust (Schistocerca gregaria), the activity of the phosphatase in the flight muscle increased during the first 3 days after the final moult. 5. The activity of glucose 6-phosphatase from insect and avian muscle was separated from that of non-specific phosphatase on a Bio-Gel P-100 column. 6. For the activities from 63 muscles, there was a strong positive correlation between those of glucose 6-phosphatase and hexokinase, but no correlation between the activities of glucose 6-phosphatase and fructose bisphosphatase. It is suggested that the role of glucose 6-phosphate in muscle is either to produce glucose from glucose 6-phosphate derived from glycogen or to provide the enzymic basis for a substrate ("futile") cycle between glucose and glucose 6-phosphatase in muscle to improve the sensitivity of the mechanism that regulates the rate of glucose phosphorylation. PMID- 6275856 TI - Dephosphorylation of oestradiol nuclear receptor in vitro. A hypothesis on the mechanism of action of non-steroidal anti-oestrogens. AB - We show that a mouse uterus nuclear phosphatase exists that is capable of inactivating nuclear oestrogen receptor complexed to oestradiol-17 beta in vitro but is ineffective when the receptor is complexed with the two non-steroidal anti oestrogens, nafoxidine and tamoxifen. We suggest that the long half-life of the tamoxifen-receptor complex versus the short half-life of the oestradiol-receptor complex in uterine nuclei in vivo is the result of the ineffectiveness of the phosphatase in dephosphorylating the anti-oestrogen-receptor complex. PMID- 6275857 TI - The insulin-stimulated cyclic AMP phosphodiesterase binds to a single class of protein sites on the liver plasma membrane. AB - The peripheral cycle AMP phosphodiesterase from rat liver plasma membranes binds with high affinity (2.4 nM) to a single class of receptor sites on the liver plasma membrane. These receptor sites appear to be proteins, as they are trypsin- and heat-labile. The sensitivity of these sites to denaturation by trypsin and heat is a first-order process. The presence of Ca2+ (5 mM) increases the affinity of these sites for the enzyme, but does not alter their total number. The receptor sites and the cyclic AMP phosphodiesterase occur in similar numbers, at around 2 pmol/mg of plasma-membrane protein. It is proposed that the peripheral, liver plasma-membrane cyclic AMP phosphodiesterase is attached to a specific site on the insulin receptor and that the binding of insulin to the receptor site triggers a conformational change in the enzyme such that the enzyme can be phosphorylated and activated by an endogenous cyclic AMP-dependent protein kinase. PMID- 6275858 TI - Mechanism of action of the cardiac glycosides on the heart. PMID- 6275859 TI - Altered states of cardiac beta-adrenoreceptors. PMID- 6275860 TI - Increases in cyclic AMP levels in rat brain regions in vivo following isoproterenol. PMID- 6275861 TI - Titers of antibodies to RANA in rheumatoid arthritis and normal sera. Relationship to Epstein-Barr virus infection. AB - The disease specificity of antibodies to rheumatoid arthritis nuclear antigen (RANA) was examined by comparing anti-RANA titers in sera from 100 patients with rheumatoid arthritis (RA) with sera from 93 healthy controls. Anti-RANA antibodies were found in 86% of the RA sera and 56% of the controls. The higher titers in the RA sera were unrelated to clinical features or to measurements of circulating immune complexes or rheumatoid factors. To study the relationship of these antibodies to previous Epstein-Barr virus (EBV) infection, antibodies to the EB virus capsid antigen (VCA) were examined and found in 94% of the RA sera and 97% of the adult controls. Four of the six RA sera without anti-VCA antibodies had detectable anti-RANA antibodies, so that we might suggest anti RANA can arise in the absence of EBV infection. From absorption experiments with non-EBV transformed extracts, we inferred that high anti-RANA titers could be due to reactions with non-Epstein-Bar virus related nuclear antigens. These data cast doubt on current speculation about a possible pathogenic role for Epstein-Barr virus in this disease. PMID- 6275862 TI - Evidence of a generalized metabolic defect in patients with hereditary chondrocalcinosis. Increased inorganic pyrophosphate in cultured fibroblasts and lymphoblasts. AB - A gene in the heterozygous state appears responsible for a 2-fold increase in pyrophosphate content of both fibroblasts and lymphoblasts cultured from patients who have dominantly inherited chondrocalcinosis. Cells from unaffected family members of this large kindred showed a pyrophosphate content in the same range as was found in unaffected, unrelated controls. Similar cells from individuals homozygous for the gene would be useful in delineating the precise biochemical abnormality responsible for the increased pyrophosphate content. PMID- 6275863 TI - Release of pyrophosphate by normal mammalian articular hyaline and fibrocartilage in organ culture. AB - Calcium pyrophosphate dihydrate crystals are found most frequently in fibrocartilaginous tissue and to a lesser extent in hyaline articular cartilage. Previous investigators found that pyrophosphate (PPi) was released into the medium by immature rabbit hyaline cartilage and osteoarthritic human cartilage in organ culture but not by normal human or mature rabbit cartilage. By employing a sensitive fluorometric assay for PPi and correcting for hydrolysis of PPi during the incubations, we detected PPi release by all normal mammalian cartilage studied. PPi release per mg wet weight of lapine and canine cartilage was paralleled by uronic acid production. Meniscal fibrocartilage, the most common site of calcium pyrophosphate deposits, also elaborate PPi. PMID- 6275864 TI - [Viral hepatitis: serological study in Puerto Rico, review and literature survey- 1981]. PMID- 6275865 TI - [Physiology of the cortical dendrites]. PMID- 6275866 TI - [Liver neoplasms in children]. AB - Presentation is made of 27 cases of liver neoplasms in infancy and childhood that were admitted to the Pediatric Hospital of the National Medical Center of the IMSS, through a period of 15 years. Sixteen cases were diagnosed as hepatoblastoma, 2 as hepatocellular carcinoma, 4 as mesenchymal hamartoma, 2 as hemangioendothelioma of infantile type, 2 were embryonal rhabdomyosarcoma of the intrahepatic bile-ducts and one case of multiple bile-ducts hamartoma. The hepatoblastoma and the mesenchymal hamartoma are neoplasms that appear more frequently before the age of 2 years; the hepatocellular carcinoma, after 5 years of age and the infantile hemangioendothelioma generally appears before 6 months old. In all, benign and malignant cases, the clinical manifestations were non specific and did not help to establish the diagnoses. The mesenchymal hamartomas were characterized by the fast growth that led to think in neoplasms of malignant nature. The selective treatment is lobectomy with a better prognosis if the neoplasm is located in the left lobe. The right lobe was the most frequently affected by the benign tumors, as well as by the malignant, and the lung was the organ wherein metastases more often were found. PMID- 6275867 TI - Intracerebroventricular administration of nanogram amounts of beta-endorphin and Met-enkephalin causes retrograde amnesia in rats. AB - The intracerebroventricular (icv) administration of 5.0 or 25.0 ng of beta endorphin or Met-enkephalin causes retrograde amnesia for a shuttle avoidance task ion rats. In both cases, the higher dose was more effective than the lower one. The present results confirm previous similar findings obtained using systemic administrations of these compounds, and suggest that the amnestic effect of beta-endorphin and Met-enkephalin is mediated centrally. PMID- 6275868 TI - [Behavior of DNCB and tuberculin skin tests in relation to the various histological types of pulmonary cancer]. PMID- 6275869 TI - [Activity of 2',3'-cyclic nucleotide 3'-phosphohydrolase in the central nervous system of rat fetal alcohol syndrome during development (author's transl)]. PMID- 6275871 TI - Effect of hypocarbia and hypercarbia on the antagonism of pancuronium-induced neuromuscular blockade with neostigmine in man. AB - The effects of variations in carbon dioxide concentration on the antagonism of pancuronium-induced neuromuscular block by neostigmine were studied in 21 patients: normocarbia (PE'CO2 5.4%, PaCO2 4.93 kPa, n = 7), hypocarbia (PE'CO2 3.6%, PaCO2 3.30 kPa, n = 7) and hypercarbia (PE'CO2 7.5%, PaCO2 7.13 kPa, n = 7). Mechanical and electromyographic responses to ulnar nerve stimulation (0.1 Hz and 2 Hz) were recorded. A 90% block during nitrous oxide in oxygen anaesthesia was maintained by incremental single injections of pancuronium and reversed with neostigmine 0.35 mg kg-1 with atropine 0.0175 mg kg-1. The recovery of twitch tension up to 50% was similar in all groups but thereafter slower in the hypercarbia group. The recovery times from 25% to 75% twitch tension correlated with PaCO2 (r = 0.55, P less than 0.05). A residual block of about 10% was seen in hypercarbic patients. However, the recovery of e.m.g. amplitude and train-of four ratios was similar in all groups. Thus, the impaired recovery of twitch tension seems to be the result of depressed contractility rather than failure of neuromuscular transmission. PMID- 6275870 TI - Pyrophosphate heart scan in patients with progressive systemic sclerosis. AB - Scintigraphic examination of the myocardium, using 99mTc-labelled pyrophosphate, was carried out in 17 patients suffering from systemic sclerosis. This connective tissue disorder very often affects the myocardium secondarily. The results of the cardiac scan were compared with the information obtained from the electrocardiogram of systolic time intervals. In addition, spirometry was undertaken to detect a potential relation between cardiac and pulmonary involvement. The scan was found to be positive in seven patients and electrocardiographic findings were pathological in five patients only. The systolic time intervals were abnormal in three patients only. A ventilation disturbance was recorded in 10 patients. No clear relation was found between the results of the individual examinations. It is concluded that pyrophosphate heart scintigraphy may detect myocardial impairment in some cases of systemic sclerosis before it is manifested by heart failure. Examination of systolic time intervals is of little importance. PMID- 6275872 TI - Antagonism of pancuronium in renal failure: no recurarization. AB - Neuromuscular transmission was measured using train-of-four stimulation, during and after anaesthesia, in 20 patients with end-stage renal failure. Neuromuscular blockade was provided with pancuronium in single doses of either 3 or 6 mg per 70 kg, and antagonized at 10% recovery with atropine and neostigmine 2.5 mg per 70 kg. Reversal was followed by progressive recovery of muscle twitch in every patient during the 3 h of the study. Recovery was more rapid after the smaller dose of pancuronium and was inversely correlated with the duration of blockade. It is concluded that, when pancuronium is antagonized with neostigmine in patients with renal failure, neuromuscular transmission recovers without evidence of recurarization. However, when large doses of pancuronium are antagonized with neostigmine 2.5 mg, recovery may be insufficient to ensure normal ventilatory function. PMID- 6275873 TI - Ontogenetic variation in rat liver, lung and kidney monooxygenase induction by low doses of benzo(A)pyrene and cigarette-smoke condensate. AB - The specific lung-AHH induction, which we previously observed after the inhalation of cigarette smoke, is not due to the route followed by the inhaled smoke, for the same phenomenon occurs after i.p. injection of either cigarette smoke condensate (CSC) or benzo(a)pyrene in low doses. In this respect lung AHH behaves completely differently from the liver and kidney enzyme, in which organs, basal AHH activity (which is low in the foetus) increases rapidly after birth to reach the adult level 2 months later, and is only inducible by CSC and low doses of BP in unweaned rats. In the lung, the basal AHH activity (low in the foetus) increases abruptly at birth, peaks in 5-day-old rats and then decreases slightly. Contrary to enzyme activity in other tissues, lung AHH cannot be induced in unweaned young animals. The enzyme subsequently becomes sensitive to inducing agents and is highly inducible in 90-day-old rats. Similar behaviour occurs in 2 other enzymes linked to cytochrome P1450: ethoxycoumarin deethylase and ethoxyresorufin deethylase. The results could be related to the particular susceptibility of the lung to develop cancer after the inhalation of cigarette smoke. PMID- 6275874 TI - An ultrastructural study of the effects of asbestos fibres on cultured peritoneal macrophages. AB - The effects of crocidolite and chrysotile fibres on lavaged peritoneal macrophages have been studied by both scanning (SEM) and transmission (TEM) electron microscopy. SEM provided little information (as the surface topography did not reflect the underlying cytoplasmic organization) except that it showed that individual macrophages often partially engulfed many long fibres in a random fashion. TEM revealed the fibres in and protruding from membrane-bound vacuoles, free in the cytoplasm and penetrating the nucleus. The cellular distribution of the fibres is discussed in terms of the cytotoxic nature of the fibres and their ability to produce a selective release of enzymes from the macrophages. PMID- 6275875 TI - The pathogenesis of chronic viral hepatitis in the nude mouse and its influence on liver regeneration after partial hepatectomy. AB - The pathogenesis of chronic viral hepatitis due to infection with mouse hepatitis virus Type 1 has been followed over a period of 30 days in the nu/nu mouse. The initial histopathology is that of a focal hepatic necrosis which evolves into a chronic hepatitis with cirrhosis. The histopathology of the experimentally induced hepatitis with mouse hepatitis Type 1 is indistinguishable from that produced by the disease in nude mouse colonies due to the natural infection in the United Kingdom and also indistinguishable from the histopathology produced by the nude mouse hepatitis virus isolated in Japan. The virus produces a temporary suppression and delay in liver regeneration of nude mice which have been partially hepatectomized and this is compared to the delay in liver regeneration induced by another more virulent strain of mouse hepatitis, Type A59, in the nude mouse. PMID- 6275876 TI - Origin of crystalloid inclusions in macrophages I: studies of peritoneal macrophages after erythrocyte ingestion. AB - In an attempt to investigate the origin of crystalloid inclusions present in bone marrow macrophages of normal adult mice and in man, murine peritoneal macrophages (which normally do not contain inclusions of this nature) were studied by light and electron microscopy 24 h after 3 daily injections of various preparations of erythrocytes. The preparations included mouse erythrocytes with and without pretreatment with collagenase, human erythrocytes and human-erythrocyte ghosts. Crystalloid inclusions were only rarely seen in the peritoneal macrophages in these circumstances. The results offered little support for the idea that crystalloid inclusions are derived from the breakdown of erythrocytes. PMID- 6275877 TI - Origin of crystalloid inclusions in macrophages II: evidence for derivation from eosinophil granulocyte breakdown. AB - Peritoneal macrophages were studied by light and electron microscopy in normal adult mice 24 h after 3 daily injections of preparations of leucocytes and platelets. Crystalloid inclusions, similar to those seen in bone marrow macrophages of normal adult mice and in man, were occasionally observed in peritoneal macrophages after administration buffy-coat white cells but not after platelets. They were much more frequently seen following the ingestion of eosinophil-rich granulocytes and were almost always associated with secondary lysosomes. Energy-dispersive analysis of X-ray provided further evidence that both crystalloid inclusions and lysosomes originated from the injected granulocytes. These observations suggest that crystalloid inclusions in marrow macrophages are derived from granulocyte breakdown and that in this respect eosinophil granulocytes are of prime importance. PMID- 6275878 TI - Unusual intracytoplasmic immunoglobulin inclusions in chronic lymphocytic leukaemia. AB - Unusual intracytoplasmic immunoglobulin inclusions were found by immunofluorescence in three patients with chronic lymphocytic leukaemia. The inclusions contained the same immunoglobulin chains as those detected on the plasma membrane, except for delta chains which were expressed on the cell surface and not in the cytoplasmic inclusions. The cytoplasmic staining persisted throughout culture for 8 or more days. An initial study of patients 1's cells showed that the inclusions contained only mu chains, and kappa chains gradually became apparent after in vitro culture. In a second study, the fresh lymphocytes contained both mu and and kappa chains. Initially, biosynthetic experiments showed production of mu chains which polymerized in the cytoplasm and were not secreted. Subsequently there was synthesis of heavy and light chains which assembled into monomeric subunits that were retained and secretion of free light chains. The apparent molecular weight of these immunoglobulin chains was larger than that of their secretory counterparts. Immunoelectronmicroscopy revealed cytoplasmic mu chains in strands of endoplasmic reticulum. In the two other patients, immunofluorescence displayed unusual staining patterns of bright networks in perinuclear areas. PMID- 6275879 TI - Structural similarities between human receptors for somatomedin C and insulin: analysis by affinity labeling. AB - Human placental receptors for insulin and somatomedin C (Sm-C) were affinity labeled with [125I]insulin and [125I]Sm-C by using the bifunctional cross-linking agent disuccinimidyl suberate. Analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis demonstrated that both labeled hormones were specifically cross-linked to three protein species with apparent molecular weights of 240 000, 310 000, and 330 000. Following disulfide bond reduction, subunits of approximately 140 000 daltons were evident. Partial reduction of disulfide bonds yielded intermediate-sized species with apparent molecular weights of 180 000, suggesting the existence of an additional, smaller subunit attached to the 140 000-dalton subunit. Limited proteolysis of the hormone-receptor complexes with chymotrypsin, trypsin, and Staphylococcus aureus V-8 protease gave similar but not identical results for each labeled receptor. The distinction between the two receptors was further documented by inhibition of affinity labeling with graded amounts of the native hormones. These data demonstrate a substantial structural similarity between the human Sm-C and insulin receptors paralleling the homology of the native hormones and their actions. PMID- 6275880 TI - 23,24,25-Trihydroxyvitamin D3, 24,25,26-trihydroxyvitamin D3, 24-keto-25 hydroxyvitamin D3, and 23-dehydro-25-hydroxyvitamin D3: new in vivo metabolites of vitamin D3. AB - Four new in vivo metabolites of vitamin D3 were isolated from the blood plasma of chicks given large doses of vitamin D3. The metabolites were isolated by methanol chloroform extraction and a series of chromatographic procedures. By use of mass spectrometry, ultraviolet absorption spectrophotometry, and specific chemical reactions, the metabolites were identified as 23,24,25-trihydroxyvitamin D3, 24,25,26-trihydroxyvitamin D3, 24-keto-25-hydroxyvitamin D3 and 23-dehydro-25 hydroxyvitamin D3. PMID- 6275881 TI - Interaction of saccharide haptens with myeloma proteins. A 270-MHz proton nuclear magnetic resonance study. PMID- 6275882 TI - Mechanism of action of methoxatin-dependent alcohol dehydrogenase. PMID- 6275883 TI - Modification of the allosteric activator site of Escherichia coli ADP-glucose synthetase by trinitrobenzenesulfonate. AB - Limited modification of Escherichia coli B ADP-glucose synthetase (EC 2.7.7.27) by trinitrobenzenesulfonate (TNBS) appeared to affect primarily the allosteric properties of the enzyme. There was little loss of the catalytic activity assayed in the absence of activator. However, the abilities of fructose 1,6-bisphosphate or hexanediol 1,6-bisphosphate to activate the enzyme, or of 5'-adenylate to inhibit the enzyme, were rapidly lost upon trinitrophenylation. Modification progressively decreased the affinity for activator, decreased the Vmax at saturating concentrations of activator, and decreased the cooperativity among activator binding sites. These effects could be completely prevented by the presence of allosteric effectors during reaction with TNBS, although a low amount of trinitrophenylation still occurred. Substrates partially protected the enzyme from reaction with TNBS. The lysyl epsilon-amino side chain was modified by trinitrophenylation, but the target was not primarily the same residue which could form a Schiff base with pyridoxal phosphate, another activator of the enzyme. A large peptide containing most of the trinitrophenyl residue was isolated after cleavage of the enzyme and was identified as part of the N terminal amino acid sequence. The migration of the enzyme on polyacrylamide gel electrophoresis or on agarose column chromatography was unchanged by modification. However, the ability of fructose-1, 6-P2 to induce the oligomerization of a mutant form of the enzyme was completely prevented by trinitrophenylation. This effect could be protected against by the presence of activator or inhibitor during reaction with TNBS. PMID- 6275884 TI - Resonance Raman spectra of cytochromes c and b in Paracoccus denitrificans membranes: evidence for heme--heme interactions. AB - Resonance Raman (RR) scattering from cytochromes b and c in the bacterium Paracoccus denitrificans was recorded by exciting with the 568.2-, 530.9-, and 520.8-nm lines of a Kr+ laser. The main features of the spectra were similar to those of the analogous cytochrome b-c1 complex derived from pigeon breast mitochondria. Differences in the 1300-cm-1 region were interpreted in terms of marker bands for heme type and spectral coupling of the hemes on the membranes. It is difficult to explain the results without invoking sharing of electronic and vibrational wave functions among the hemes. This conclusion documents the potential to study the physics of electron transport in functioning membrane by monitoring the RR spectra. PMID- 6275885 TI - Protein kinases of retinal rod outer segments: identification and partial characterization of cyclic nucleotide dependent protein kinase and rhodopsin kinase. AB - Protein kinase activity of dark-adapted bovine rod outer segments is partitioned by centrifugation into soluble and membrane-bound fractions. The soluble kinases are separated by DEAE-cellulose chromatography into three peaks of activity, which can be classified by substrate specificity and cyclic nucleotide dependence into two categories. One peak of protein kinase activity has the characteristics reported for rhodopsin kinase (category one); it phosphorylates only bleached rhodopsin, and its activity is not affected by light, exogenous adenosine cyclic 3',5'--monophosphate (cAMP), guanosine cyclic 3',5'-monophosphate (cGMP), or a protein kinase inhibitor from skeletal muscle. Rhodopsin kinase has an apparent molecular weight of 68 000. The second category of kinase includes two peaks of activity which are stimulated severalfold by cAMP or cGMP but not by light. These protein kinases phosphorylate soluble proteins including histones and a protein kinase substrate prepared from rat intestine but not rhodopsin. The two peaks elute from DEAE-cellulose with 0.09 and 0.20 M KCl, suggesting that they are similar respectively to type I and type II cyclic nucleotide dependent protein kinases that have been characterized in other tissues. The activity of type I kinase is variable and much less than that of the type II enzyme; its molecular weight was not determined. The type II protein kinase has an apparent molecular weight of 165 000. This study confirms that different protein kinase enzymes catalyze selectively the phosphorylation of bleached rhodopsin and soluble proteins, and it repudiates the speculation in a previous publication [Farber, D. B., Brown, B. M., & Lolley, R. N. (1979) Biochemistry 18, 370-378] that a single protein kinase might catalyze both phosphorylation reactions. PMID- 6275886 TI - Synthesis of disaturated phosphatidylcholine by cholinephosphotransferase in rat lung microsomes. AB - 1. Incubation of rat lung microsomes with cytidine diphospho[methyl-14C]choline resulted in synthesis of radioactively labeled phosphatidylcholine. 2. 10-15% of this phosphatidylcholine appeared to be disaturated species. In similar experiments with rat liver microsomes only 2-3% of the radioactivity was present in the disaturated species. 3. When de novo synthesis was blocked after 5 min by addition of Ca2+ no increase in the proportion of disaturated phosphatidylcholine was observed upon further incubation of lung microsomes. Under these conditions the enzymes involved in a remodeling mechanism, i.e. phospholipase A and acyl CoA: lysophosphatidyl-choline acyltransferase, remain fully active. 4. Addition of diacylglycerols from egg phosphatidylcholine containing trace amounts of di[1 14C]palmitoyl glycerol resulted in direct incorporation of 14C label into phosphatidylcholine. The rate of phosphatidylcholine synthesis measured from incorporation of di[1-14C]palmitoyl glycerol equalled that observed with labeled CDP choline. 5. These results support the conclusion that disaturated phosphatidylcholine in lung can be formed by direct utilization of disaturated diacylglycerol and is not produced exclusively via remodelling of de novo synthesized unsaturated species. PMID- 6275887 TI - Isolation and identification of tachysterol3 in rat skin exposed to ultraviolet light. AB - In order to investigate the mechanism of the cutaneous synthesis of vitamin D3 in vivo, a photolysis by-product of previtamin D3 in organic solvent, namely, tachysterol3 was isolated and identified in ultraviolet-exposed rat skin. Tachysterol3 was identified by chromatographic comigration of an ultraviolet absorbing peak wih authentic tachysterol3 in straight-phase HPLC (Zorbax SIL column) and reversed-phase HPLC (Nucleosil 5C18 column), and by the ultraviolet absorption spectrum. The comparison of the extent of formation of tachysterol3 between in rat skin in vivo and in ethanol solution indicated that the yield in rat skin is similar to that in organic solvent. The results suggest that the reaction mechanism in rat skin in vivo may be identical wit that in organic solvents. PMID- 6275888 TI - Effect of thyroid state on ketogenic capacity of the isolated perfused liver of starved rats. AB - The effect of different thyroid states on the oxidation of free fatty acids was investigated in the isolated perfused liver of 24-h-starved rats. 1. Compared with the euthyroid control the oxidation of oleate to ketone bodies as well as to CO2 was increased in hyper-, while it was unchanged in hypothyroid livers. 2. The addition of carnitine stimulated oleate oxidation in livers from eu- and hyperthyroid rats, but was without effect in hypothyroid livers. 3. Glucose did not affect the thyroid hormone-mediated effect of oleate conversion to ketone bodies. 4. Hepatic oxidation of octanoate was similar in all thyroid states. 5. Re-esterification of oleate was enhanced in hypo-, but reduced in hyperthyroidism. 6. The concentration of hepatic malonyl-CoA was decreased in hypo- and unchanged in hyperthyroid livers. 7. The concentration of cyclic AMP was elevated in the liver of hyperthyroid rats, no differences were observed between eu- and hypothyroid livers. However, increasing the hepatic cyclic AMP content by the addition of glucagon did not stimulate ketogenesis in eu- and hypothyroid livers. 8. The results indicate that thyroid hormones stimulate oleate oxidation by an accelerated transport of its CoA derivative into the mitochondrial compartment. PMID- 6275889 TI - ATP synthesis during exogenous NADH oxidation. A reappraisal. AB - This paper reports a reinvestigation on the pathway for mitochondrial oxidation of exogenous NADH and on the related ATP synthesis, first reported 30 years ago (Lehninger, A.L. (1951) J. Biol. Chem. 190, 345-359). NADH oxidation, both in intact and in water-treated mitochondria, is 90% inhibited by mersalyl, an inhibitor of the outer membrane NADH-cytochrome b5 reductase, and 10% inhibited by rotenone. The mersalyl-sensitive, but not the rotenone-sensitive, portion of NADH oxidation is stimulated by exogenous cytochrome c. Part of ATP synthesis is independent of exogenous NADH and cytochrome c, and is inhibited by rotenone and antimycin A, and is therefore due to oxidation of endogenous substrates. Another part of ATP synthesis is dependent on exogenous NADH and cytochrome c, is insensitive to rotenone and antimycin A, and is due to operation of cytochrome oxidase. It is concluded that (i) oxidation of exogenous NADH in the presence of cytochrome c proceeds mostly through NADH-cytochrome b5 reductase and cytochrome b5 on the outer membrane and then through cytochrome oxidase via the cytochrome c shuttle, and (ii) ATP synthesis during oxidation of exogenous NADH is partly due to oxidation of endogenous substrates and partly to operation of cytochrome oxidase receiving electrons from the outer membrane via cytochrome c. PMID- 6275890 TI - Affinity chromatography purification of cytochrome c oxidase and b-c1 complex from beef heart mitochondria. Use of thiol-sepharose-bound Saccharomyces cerevisiae cytochrome c. AB - A method for simultaneous purification of cytochrome c reductase and cytochrome c oxidase using a cytochrome c affinity column is presented. Cytochrome c from Saccharomyces cerevisiae was linked to an activated thiol-Sepharose gel via its Cys-102 residue located far from the lysine residues on the front side of the molecule, responsible for the interaction with the reductase and oxidase. In previously reported affinity chromatography techniques these lysine residues most probably reacted with the column. Cytochrome c oxidase and reductase from bovine heart mitochondria bind specifically to the affinity column and can be recovered separately at different ionic strength in the elution buffer. The enzymes are highly pure and active. PMID- 6275891 TI - EPR studies of the photodissociation reactions of cytochrome c oxidase-nitric oxide complexes. AB - Three complexes of NO with cytochrome c oxidase are described which are all photodissociable at low temperatures as measured by EPR. The EPR parameters of the cytochrome a2+(3)-NO complex are the same both in the fully reduced enzyme and in the mixed-valence enzyme. The kinetics of photodissociation of cytochrome a2+(3)-NO and recombination of NO with cytochrome a2+(3) (in the 30-70 K region) revealed no differences in structure between cytochrome a2+(3) in the fully reduced and the mixed-valence states. The action spectrum of the photodissociation of cytochrome a2+(3)-NO as measured by EPR has maxima at 595, 560 and 430 nm, and corresponds to the absorbance spectrum of cytochrome a2+(3) NO. Photodissociation of cytochrome a2+(3)-NO in the mixed-valence enzyme changes the EPR intensity at g 3.03, due to electron transfer from cytochrome a2+(3) to cytochrome a3+. The extent of electron transfer was found to be temperature dependent. This suggests that a conformational change is coupled to this electron transfer. The complex of NO with oxidized cytochrome c oxidase shows a photodissociation reaction and recombination of NO (in the 20-40 K region) which differ completely from those observed in cytochrome a2+(3)-NO. The observed recombination occurs at a temperature 15 K lower than that found for the cytochrome a2+(3)-NO complex. The action spectrum of the oxidized complex shows a novel spectrum with maxima at 640 and below 400 nm; it is assigned to a Cu2+B-NO compound. The triplet species with delta ms = 2 EPR signals at g 4 and delta ms = 1 signals at g 2.69 and 1.67, that is observed in partially reduced cytochrome c oxidase treated with azide and NO, can also be photodissociated. PMID- 6275892 TI - Partial purification of an intercalated disc-containing cardiac plasma membrane fraction. AB - The sarcolemma of cardiac muscle cells contains a specialised junctional region, the intercalated disc which includes three types of intercellular junction, the macula and fascia adherens and the nexus or gap junction. To facilitate the isolation of these junctions a procedure for the partial purification from mouse hearts of a subcellular fraction containing the intercalated disc region of the sarcolemma was developed. This involved investigating methods of tissue disruption that preserve the integrity of the intercalated disc and minimise myofibrillar entrapment of organelles. Examination of the distribution of marker enzymes showed that relative to the homogenate the intercalated disc fraction prepared by sucrose density centrifugation was only enriched 1.5- to 3-fold in 5' nucleotidase and (Na+ + K+)-ATPase activities, whereas mitochondrial and sarcoplasmic reticulum marker enzymes were low. The properties of the intercalated disc-containing fraction were compared with the vesicular sarcolemmal fractions devoid of junctional complexes prepared by other methods. PMID- 6275893 TI - Fluid phase activation of proenzymic C1r purified by affinity chromatography. AB - 1. Proenzymic C1r was purified from human plasma in a two-step technique involving indirect affinity chromatography on Sepharose Ig anti-C1s. The capacity of C1r to monomerize at pH 5.0 and to redimerize at neutral pH was used for selective elution of C1r. The yield in purified C1r was 39% from plasma; no trace of contaminating serine proteases was detected from [3H]diisopropyl phosphorofluoridate labelling of C1r. 2. C14 was able to undergo a two-way autoactivation: an intramolecular catalytic process catalysed by proenzymic C1r itself and an intermolecular reaction catalysed by activated C1r formed in the process of the reaction. DFP (5mM) and C1 Inh at a C1 Inh/C1r ratio of 1:1 were effective on the solely intermolecular activation, leading to partial inhibition of the autoactivation from proenzymic C1r: C1r formed during the activation was titrated by the inhibitors. Calcium, high ionic strength or acid pH decreased C1r activation. The pH effect was characterized by a slowed-down reaction below pH 6.0 and no net influence at values as high as 10.5. The two types of activation developed similarly as a function of pH. 3. Peripheral iodination of C1r revealed differences in label distribution between proenzymic (A chain moiety 48%, B chain moiety 52%) and activated C1r (A chain 20%, B chain 80%). Two different conformational states of C1r were also suggested by 125I-labelling at different temperatures. PMID- 6275894 TI - The affinity of rabbit muscle fructose-bisphosphatase for fructose bisphosphate. AB - This paper reports that microM concentrations of fructose bisphosphate are titrated by rabbit muscle fructose-bisphosphatase (D-fructose-1,6-bisphosphate 1 phosphohydrolase, EC 3.1.3.11) when the enzyme concentration is varied in the range which secures measurable initial velocities of reaction: a result that can only be explained by supposing that the enzyme has a greater affinity for fructose bisphosphate than suggested by Fernando, J., Enser, M., Pontremoli, S. and Horecker, B.L. (1968) Arch. Biochem. Biophys. 126, 599-606. The results also suggest that the keto form of the substrate may be the preferred configuration and that the enzyme is inhibited by magnesium-bound fructose bisphosphate. PMID- 6275895 TI - The EPR spectra of copper transferrin complexes at 2-4 GHZ. PMID- 6275896 TI - Spectral and other studies on the intestinal haem receptor of the pig. AB - We recently demonstrated the presence of a Triton-solubilized high-affinity haem binder on the pig duodenal brush border membrane. The association of haem to the binding factor was determined using radioactive haem and is now studied by a spectrophotometric technique. The binding alters the Soret absorption band of haem from 395 nm to 413 nm. The dissociation constant for the binding of haem to the solubilized binding factor was estimated to be about 10(-9) M by difference spectroscopy. Human serum albumin could not prevent the solubilized binding factor from binding haem. Trypsin digestion destroyed the binder. PMID- 6275897 TI - The effect of guanidine hydrochloride on the conformation of bovine pancreatic DNAase I as measured with circular dichroism. AB - The denaturation of bovine pancreatic DNAase I (EC 3.1.21.1) by guanidine hydrochloride (GdnHCl) has been investigated with circular dichroism in the presence and absence of 1 mM Ca2+ at the wavelength region of 210-240 nm at 12.25 and 36 degree C. The change of the molar ellipticity at 220 nm by GdnHCl titration showed cooperative transition at each temperature and the midpoints of the titrations occurred near 2 M GdnHCl. At each temperature, the denaturation of DNAase I in the presence of 1 mM Ca2+ occurred a little slowly as compared with that in the absence of Ca2+. This suggests that 1 mM Ca2+ can to some extent stabilize the secondary structure of DNAase I against GdnHCl denaturation. The apparent free energy for the denaturation of DNAase I obtained by GdnHCl titration was calculated as 9.3 +/- 0.3 kcal/mol and 8.9 +/- 0.2 kcal/mol at 25 degree C in the presence and absence of 1 mM Ca2+, respectively. The possible regions for the alpha -helix and beta -structure of DNAase I were predicted from the amino acid sequence by probability calculation of Chou, P.Y. and Fasman, G.D., Adv. Enzymol. 47, 45-148. The characteristic feature is that the NH2 terminal half of DNAase I is rich in beta -structure and the COOH-terminal half contains mainly alpha -helix. PMID- 6275898 TI - The reaction of the trifluoromethylphenylcarbamylated lysine-13 derivative of horse cytochrome c with cytochrome oxidase. AB - The kinetics of oxidation of horse cytochrome c and the trifluoromethylphenylcarbamylated lysine-13 derivative by cytochrome c oxidase (ferrocytochrome c: oxygen oxidoreductase, EC 1.9.3.1) were compared using both spectrophotometric and polarographic methods under different experimental conditions. The rate constants measured spectrophotometrically in 0.025 M tris cacodylate buffers were similar with the two cytochrome at pH 7.8, but those with the derivative were slightly higher at pH 6. Rates measured with polarographic assays in these buffers were the same with the horse and the derivative cytochromes c at pH 6, but at pH 7.8 the rates with the derivative were less at cytochrome c concentrations between 0.05 and 0.5 micro M and were greater at higher concentrations. The pH optima in the polarographic assays of the derivative and the native pigments were different in 0.025 M Tris-cacodylate buffers; in spectrophotometric assays at pH 7.8 the trifluoromethylphenylcarbamylated lysine-13 cytochrome c showed a greater sensitivity to changes in ionic strength than did the native cytochrome. The variations in apparent Km and V values calculated from spectrophotometric and polarographic assays with the two cytochromes cannot be explained as due to changes in binding of cytochrome c to cytochrome oxidase. The large excess of O2 uptake seen in polarographic assays with horse cytochrome c over that expected from spectrophotometric measurements was not apparent with the trifluoromethylphenylcarbamylated lysine-13 derivative. Thus, the derivative seems to have decreased ability to form the combination of cytochrome c with the oxidase giving high turnover rates. PMID- 6275899 TI - Presence and characterization of two protein kinase activities in human seminal fluid. AB - The presence of two protein kinase (ATP: protein phosphotransferase, EC 2.7.1.37) activities has been demonstrated in human seminal fluid, utilizing partially dephosphorylated phosvitin and lysine-rich histones as model acceptor substrates. Both kinase activities were maximal in the presence of MgCl2 and a sulfhydryl protecting agent such as dithiothreitol; however, the histone kinase was stimulated to a greater extent by the latter. The histone kinase displayed a broad shoulder of activity at pH values of 7.1 to 7.6 with optimal activity at pH 8.0, and was inhibited by increased ionic strength (53% at 160 mM NaCl) and by the cyclic AMP-dependent protein inhibitor from rabbit muscle. The kinase activity towards phosvitin exhibited a broad pH profile with maximal activity at pH 7.2, was slightly stimulated by NaCl (20% at 160 mM), and was unaffected by the cyclic AMP-dependent protein kinase inhibitor. Kinetic studies revealed more than one apparent Km for the protein substrates and ATP. These differences in enzymic properties of kinase activities towards phosvitin and lysine-rich histones strongly indicate the presence of multiple enzymes. It appears that the histone kinase activity is attributable to the free catalytic subunit of a cyclic AMP-dependent enzyme. The protein kinase activities of seminal fluids from vasectomized men were 12-20% of those found for seminal fluids of normal men. This suggests that sperm may be a major source of protein kinase activities in seminal fluid. PMID- 6275900 TI - The site of histone H2b phosphorylated by a cyclic nucleotide independent histone kinase. AB - A cyclic nucleotide independent histone kinase was demonstrated in bovine thymus extract. This enzyme was very similar to that found previously in the cytoplasm and nucleus of human tonsillar lymphocytes (Farago et al., 1973, Biochim. Biophys. Acta 297, 517 and 1974, ibid 370, 459). High-performance liquid chromatography of the tryptic phosphopeptides of calf thymus histones H1 and H2b phosphorylated by the histone kinase or by the catalytic subunit of cylic AMP dependent protein kinase showed that the intrinsic substrate specificity of these enzymes differed significantly. Under our experimental conditions the histone kinase phosphorylated preferentially the Ser-32 residue, and it did not phosphorylate the Ser-36 residue of histone H2b, while Ser-36 was phosphorylated preferentially by the cyclic AMP dependent protein kinase. A peptide containing the amino acid sequence of histone H2b from Gly-26 to Lys-34 (Gly-Lys-Lys-Arg-Lys Arg-Ser-Arg-Lys-Ala) was synthesized. This peptide was a competitive inhibitor of histone H1 phosphorylation by the histone kinase and it was also a substrate for this enzyme. PMID- 6275901 TI - Diethylstilbestrol treatment increases the amount of choline kinase in rooster liver. AB - Studies have been performed on the mechanism by which diethylstilbestrol stimulates the activity of choline kinase in livers from cockerels. The enzyme was purified 700-900-fold by affinity chromatography. The increased enzyme activity could not be accounted for by diethylstilbestrol alteration of the kinetic constants of the enzyme. Rabbit antibody was raised to the purified enzyme. Titration studies with antiserum demonstrated a 2-fold increase in the amount of choline kinase in diethylstilbestrol-treated cytosol, which correlated with a 2-fold elevation of the activity of the enzyme. We conclude that diethylstilbestrol stimulates the activities of choline kinase in cockerel liver by corresponding increase in the amount of enzyme. PMID- 6275902 TI - Differential regulation of calcium-dependent and calcium-independent cyclic nucleotide phosphodiesterases from heart by palmitoylcarnitine and palmitoyl coenzyme A. AB - Regulation of Ca2+-dependent (peak I) and Ca2+-independent (peak II) phosphodiesterases from the heart by various fatty acyl esters and phospholipids were studied. DL-Palmitoylcarnitine stimulated the basal activity (in the absence of Ca2+) of peak I enzyme, while non-competitively inhibiting peak II enzyme with respect to cyclic AMP. It had no effect on other species of Ca2+-independent phosphodiesterases, including cyclic AMP- and cyclic GMP-specific enzymes from the lung, and cyclic CMP enzyme from the liver Palmitoyl-CoA and phosphatidylserine also stimulated the basal activity of peak I enzyme, but they were without effect on peak II enzyme. In comparison, DL-palmitoylcarnitine inhibited Ca2+-dependent activity of cardiac myosin light chain kinase, whereas phosphatidylserine was without effect. It is conceivable that differential regulation of phosphodiesterases by these lipids could profoundly alter the levels or effects, or both, of cyclic nucleotides and Ca2+ in the myocardium. PMID- 6275903 TI - Evidence for selective regulation of the phosphorylation of myocyte proteins by isoproterenol and prostaglandin E1. AB - Both isoproterenol and prostaglandin E1 increased the activation state of cyclic AMP-dependent protein kinase in cultured myocytes; however, only isoproterenol enhanced phosphorylase activity and contractile state. Following the incubation of intact myocytes with 32PO3-(4), 32 phosphoproteins were resolved from total cellular proteins by electrophoresis in sodium dodecyl sulfate polyacrylamide gels followed by autoradiography. Isoproterenol stimulated 32PO3-(4) incorporation into 16 proteins, including 2 phosphoproteins not observed under control conditions. By contrast, prostaglandin E1 neither caused a measurable change in the protein phosphorylation pattern nor interfered with isoproterenol's capacity to do so. Isoproterenol stimulated myocyte protein phosphorylation in either the presence or absence of extracellular Ca2+. The results suggest that the regulation of protein phosphorylation following adenylate cyclase stimulation is: (1) an agonist-specific process and not due solely to a random accumulation of intracellular cycle AMP and activation of protein kinase; (2) the Ca2+ mobilization component of beta-receptor activation does not account for the paradoxical effects of isoproterenol and prostaglandin E1; (3) activation of cyclic AMP-dependent protein kinase does not always result in an enhancement of protein phosphorylation. PMID- 6275904 TI - Catecholamine binding to the alpha-adrenergic receptors of hamster adipocytes. Evidence that guanine nucleotides regulate this binding to the alpha 2-receptor subtype. AB - The binding characteristics of the alpha-component of (-)-[3H]norepinephrine to hamster adipocyte membranes were studied. Binding was rapid, equilibrium in 20 min at 25 degrees C. Dissociation of specific binding by 10 microM phentolamine suggested dissociation from two different sites. The time course of dissociation induced by a 50-fold dilution was unchanged by the addition of norepinephrine, suggesting the absence of cooperative binding sites. [3H]norepinephrine binding was saturable, yielding curvilinear Scatchard plots. Computer modeling of these data further supported the existence of two classes of binding sites, one with high affinity (KD = 23 nM) but low binding capacity (96 fmol/mg protein) and one with low affinity (KD = 400 nM) but high binding capacity (1000 fmol/mg protein). Adrenergic ligands competed with [3H]norepinephrine binding in the following order of potency: (-)-norepinephrine greater than (-)-epinephrine much greater than (+)-norepinephrine greater than (-)-isoproterenol. Displacement by the selective alpha-adrenergic drugs prazosin, clonidine, and yohimbine yielded biphasic curves consistent with binding of [3H]norepinephrine to both alpha 1-(14 22%) and alpha 2-(78-86%) receptor subtypes. Although Gpp(NH)p failed to alter the binding of [3H] dihydroergocryptine, it severely reduced the binding affinity of (-)-epinephrine, (-)-norepinephrine and the selective alpha 2-agonist, clonidine. The inhibitory effects of clonidine and of the alpha-component of (-) epinephrine on the adrenocorticotropin-stimulated cyclic AMP production in the intact adipocyte were closely correlated with their effects on the binding of both [3H]norepinephrine and [3H]dihydroergocryptine. Conversely, yohimbine but not prazosin markedly antagonised the alpha-inhibitory effect of norepinephrine on cyclic AMP production. These data led to conclude that [3H]norepinephrine can be successfully used to study the entire alpha-adrenergic receptor population of hamster fat cells and that the predominant alpha 2-receptor subtype exists in two different affinity states for agonists, the proportions of which are modulated by guanine nucleotides. PMID- 6275905 TI - Cholinergic muscarinic stimulation of steroidogenesis in bovine adrenal cortex fasciculata cell suspensions. AB - Acetylcholine was found to acutely stimulate cortisol production by bovine fasciculata adrenocortical cell suspensions. This effect was maximal at 10(-4) M acetylcholine concentration, resulted in a 5-fold increase in cortisol production over the control after 1 h incubation, and represented about one fifth of the ACTH maximal stimulation under the same conditions. Acetylcholine-stimulated steroidogenesis was concentration-dependent (10(-8)-10(-5) M), proportional to the cell number (5 X 10(5)-1 X 10(6)) and reached a plateau after 30 min incubation. Use of various cholinergic specific agonists and antagonists showed that the steroidogenic action of acetylcholine was a typical muscarinic effect. This character is in agreement with the previously demonstrated presence of muscarinic receptors in bovine adrenocortical tissue. The steroidogenic effect of acetylcholine required the presence of extracellular calcium in the medium and was impaired upon addition of tetracaine and procaine. No change in cyclic AMP nor cyclic GMP levels could be detected in the system under acetylcholine stimulation. Acetylcholine appeared to exhibit a synergistic effect in combination with ACTH, and exogenous cyclic AMP; these observations suggest a different mechanism of action for acetylcholine and ACTH and point to a possible cholinergic participation in the regulation of adrenocortical differentiated functions in vivo. PMID- 6275906 TI - Regulation of adrenergic stimulation of hepatic adenylate cyclase by divalent cations. AB - Liver plasma membrane adenylate cyclase was stimulated paradoxically by an alpha 2-adrenergic mechanism under conditions of low metal ion and low GTP concentrations. In untreated membranes, epinephrine stimulation was GTP-dependent and was mediated by beta-adrenergic receptors since it was completely blocked by propranolol, but unaffected by dihydroergocryptine. Pre-treatment of membranes to remove or reduce divalent cations and guanine nucleotides changed epinephrine stimulation to a form that was mediated by alpha 2-receptors since it was completely blocked by dihydroergocryptine, phenoxybenzamine and yohimbine, but not by propranolol or prazosin. The pre-treatment did not alter enzyme activation by isoproterenol or glucagon, alpha 2-Adrenergic stimulation of adenylate cyclase in depleted membranes required the presence in the assay of 1-2 mM Mg2+ and small amounts of exogenous GTP (less than or equal to 50 nM). Increasing the Mg2+ or GTP concentration in the assay produced a progressive reversal of epinephrine stimulated activity from an alpha 2-adrenergic form to a predominantly beta adrenergic form. Readdition of Ca2+ or Mg2+, but not Mn2+, into depleted membranes by incubation in the presence of metal reestablished the pattern of enzyme sensitivity to epinephrine to that seen with untreated membranes i.e., it changed from alpha 2- to beta-receptor mediation. Alterations in membrane and assay content of metal ions and GTP did not result in the activation of the enzyme by vasopressin or angiotensin II. These findings demonstrate the ability of Ca2+, Mg2+ and GTP to control the coupling of beta- and alpha 2-adrenergic receptors with liver adenylate cyclase. It is hypothesized that the cations act by regulating the interaction of the receptors with adrenergic agonists and/or the guanine nucleotide binding protein(s) which is postulated to be involved in control of the enzyme. PMID- 6275907 TI - Testicular membranes with improved stability of the gonadotropin receptor. AB - A plasma membrane fraction has been prepared from rat testis using an aqueous double-phase polymer system containing dextran, poly(ethylene glycol) 6000 and Zn2+. The membrane-associated gonadotropin receptor for lutropin and human choriogonadotropin can be markedly stabilized by a thawing-washing step of frozen membranes which prolongs the apparent half-life of the unoccupied membrane associated receptors from less than 1 h at 37 degrees C to greater than 5h. Also, no degradation of 125I-labeled human choriogonadotropin was detected following incubation with the membrane fraction. The equilibrium binding was characterized by an apparent association constant of 1.6 x 10(10) M-1 and a receptor content of 33 fmol/mg protein. Binding kinetics yielded as association rate constant of 1.0 x 10(8) M-1 x min-1, while the dissociation rate constant for human choriogonadotropin was too low to be accurately determined under the conditions used. In contrast, ovine lutropin could be reversibly bound to the membranes leaving the previously occupied receptors available for binding by 125I-labeled human choriogonadotropin. PMID- 6275908 TI - Development of cholinephosphotransferase in guinea pig lung mitochondria and microsomes. AB - Development of mitochondrial and microsomal choline phosphotransferase in the fetal guinea pig lung was investigated. The activity in fetal mitochondria was more than twice of that in fetal microsomes. However, in adult lung, the enzyme was distributed mostly in microsomes. In fetal lung, both the mitochondrial and microsomal enzyme activity was greatest at approx. 81% of the total gestation period (55 days). The specific activity in the microsomal fraction than declined until term, but increased again in the 24-h newborn from 1.0 to 2.3 nmol/min per mg protein. The activity in the mitochondrial fraction declined after 61 days (2.8 nmol/min per mg) to a minimal level at term (0.6 nmol/min per mg). Although the enzyme activity decreased from day 55 (1.2 nmol/min per mg), the amount of phosphatidylcholine gradually increased between day 55 and term. PMID- 6275909 TI - Oxygenation dependence of the transverse relaxation time of water protons in whole blood at high field. AB - At high and medium magnetic field, the transverse NMR relaxation rate (T-1(2)) of water proteins in blood is determined predominantly by the oxygenation state of haemoglobin. T-1(2) depends quadratically on the field strength and on the proportion of haemoglobin that is deoxygenated. Deoxygenation increases the volume magnetic susceptibility within the erythrocytes and thus creates local field gradients around these cells. From volume susceptibility measurements and the dependence of T-1(2) on the pulse rate in the Carr-Purcell-Meiboom-Gill experiment, we show that the increase in T-1(2) with increasing blood deoxygenation arises from diffusion of water through these field gradients. PMID- 6275910 TI - Dissociation of human choriogonadotropin from the pseudopregnant rat ovary as a complex to a receptor fragment during perifusion and incubation. AB - Pseudopregnant rats were injected intravenously with radioactively-labelled human choriogonadotropin (CG). The animals were killed 2 h after the injection and the ovaries, liver and kidney were subjected to perifusion. Radioactivity was released from the ovaries at an increasing rate during perifusion, mainly in a complex form with a molecular size between human CG and the solubilized receptor human CG complex. The increase in the rate of radioactivity release was inhibited by N-ethylmaleimide and CuCl2, but not by MgCl2, Trasylol, N alpha-tosyl-L phenylalanine chloromethyl ketone or N alpha-p-tosyl-L-lysine and CuCl2 chloromethyl ketone. Intact hormone dissociation from the complex at pH 3. After perifusion the ovarian tissue radioactivity only as receptor-hormone complexes. Only free radioiodine released from the control tissues, liver and kidney during perifusion. The low molecular weight hormone complex was also released from a homogenate of pseudopregnant rat ovaries prelabelled in vivo with radioactivity labelled human CG during incubation in a hypotonic medium. The release of this complex was likewise inhibited by alkylating agents and heavy metals, and intact hormone dissociated from the complex at pH 3. A similar human CG complex was released also from purified receptor-human CG complex during incubation with ovarian homogenate, and presence of N-ethylmaleimide or use of heat inactivated ovarian homogenate inhibited this process. The present results indicate that the in vivo bound human CG sheds from the luteal tissue in perifusion and incubation as a low molecular weight complex. This may be a facet in the processing and elimination of occupied LH receptors from the ovary. PMID- 6275911 TI - Purification, characterization and production of rabbit antibodies to rat liver particulate, high-affinity, cyclic AMP phosphodiesterase. AB - The cyclic nucleotide phosphodiesterase (EC 3.4.16) activities of a rat liver particulate fraction were analyzed after solubilization by detergent or by freeze thawing. Analysis of the two extracts by DEAE-cellulose chromatography revealed that they contain different complements of phosphodiesterase activities. The detergent-solubilized extract contained a cyclic GMP phosphodiesterase, a low affinity cyclic nucleotide phosphodiesterase whose hydrolysis of cyclic AMP was activated by cyclic GMP and a high affinity cyclic AMP phosphodiesterase. The freeze-thaw extract contained a cyclic GMP phosphodiesterase and two high affinity cyclic AMP phosphodiesterase, but no low affinity cyclic nucleotide phosphodiesterase. The cyclic AMP phosphodiesterase activities from the freeze thaw extract and from the detergent extract all had negatively cooperative kinetics. One of the cyclic AMP phosphodiesterases from the freeze-thaw extract (form A) was insensitive to inhibition by cyclic GMP; the other freeze-thaw solubilized cyclic AMP phosphodiesterase (form B) and the detergent-solubilized cyclic AMP phosphodiesterase were strongly inhibited by cyclic GMP. The B enzyme appeared to be converted into the A enzyme when the particulate fraction was stored for prolonged periods at -20 degrees C. The B form was purified extensively, using DEAE-cellulose, a guanine-Sepharose column and gel filtration. The enzyme retained its negatively cooperative kinetics and high affinity for both cyclic AMP and cyclic GMP throughout the purification, although catalytic activity was always much greater for cyclic AMP. Rabbit antiserum was raised against the purified B enzyme and tested via a precipitin reaction against other forms of phosphodiesterase. The antiserum cross-reacted with the A enzyme and the detergent-solubilized cyclic AMP phosphodiesterase from rat liver. It did not react with the calmodulin-activated cyclic GMP phosphodiesterase of rat brain, the soluble low affinity cyclic nucleotide phosphodiesterase of rat liver or a commercial phosphodiesterase preparation from bovine heart. These results suggest a possible interrelationship between the high affinity cyclic nucleotide phosphodiesterase of rat liver. PMID- 6275912 TI - Effects of Ca2+, calmodulin and cyclic AMP on the phosphorylation of endogenous proteins by homogenates of rt islets of langerhans. AB - Activation of Ca2+ -calmodulin- and cyclic AMP-dependent protein kinases has been suggested to be involved in stimulus-secretion coupling in the pancreatic beta cell. To study the properties of suc kinases and their endogenous protein substrates homogenates of rat islets of Langerhans were incubated with [gamma 32P]ATP. Phosphorylated proteins were separated by sodium dodecyl sulphate polyacrylamide gel electrophoresis and detected by autoradiography. The phosphorylation of certain proteins could be enhanced by Ca2+ plus calmodulin or by cyclic AMP. The major effect of Ca2+ and calmodulin was to stimulate the phosphorylation of a protein (P53) of molecular weight 53,100 +/- 500 (n = 15). Maximum phosphorylation of protein P53 occurred within 2 min with 2 micrometers free Ca2+ and 0.7 micrometers calmodulin. Incorporation of label into protein P53 was inhibited by trifluoperazine or W7 but not by cyclic AMP-dependent protein kinase inhibitor. Phosphorylation of a proteins of similar molecular weight could be enhanced to a lesser extent in the absence of Ca2+ but in the presence of cyclic AMP and 3-isobutylmethylxanthine: this phosphorylation was blocked by cyclic AMP-dependent protein kinase inhibitor. Cyclic AMP also stimulated incorporation of label into polypeptides of molecular weights 55,000 and 70 80,000. The results are consistent with the hypothesis that protein phosphorylation mechanisms may play a role in the regulation of insulin secretion. PMID- 6275913 TI - Studies on protein kinases in two human rectocolic cell lines by polyacrylamide gel electrophoresis. PMID- 6275914 TI - The effect of the administration of the adrenocorticotrophic hormone to rats on in vivo steroidogenesis and on the reduction cytochrome P-450 in adrenal mitochondria. PMID- 6275915 TI - Ionic channel density of excitable membranes can act a bifurcation parameter. AB - As the maximal K+-conductance (or K+-channel density) of the Hodgkin-Huxley equations is reduced, the stable resting membrane potential bifurcates at a subcritical Hopf bifurcation into small amplitude unstable oscillations. These small amplitude solutions jump to large amplitude periodic solutions that correspond to a repetitive discharge of action potentials. Thus the specific channel density can act as a bifurcation parameter, and can control the excitability and autorhythmicity of excitable membranes. PMID- 6275916 TI - Fetal and postnatal development of the adrenal glands in Macaca mulatta. PMID- 6275917 TI - GnRH-receptor interaction. VI. Effect of progesterone and estradiol on hypophyseal receptors for GnRH, and serum and hypophyseal concentrations of gonadotropins in ovariectomized ewes. PMID- 6275918 TI - Temperature dependence of electron transfer between bacteriopheophytin and ubiquinone in protonated and deuterated reaction centers of Rhodopseudomonas sphaeroides. AB - The rate of the electron-transfer reaction between bacteriopheophytin and the first quinone in isolated reaction centers of Rhodopseudomonas sphaeroides has an unusual temperature dependence. The rate increases about threefold with decreasing temperature between 300 and 25 K, and decreases abruptly at temperatures below 25 K. Partial deuteration of the reaction centers alters the temperature dependence of the rate constant. Qualitative features of the temperature dependence can be understood in the context of a theory of nonadiabatic electron transfer (Sarai, 1980. Biochim. Biophys. Acta 589:71-83). We conclude that very low-energy (10-50 cm-1) processes, perhaps skeletal vibrations of the protein, are important to electron transfer. Higher-energy vibrations, possibly involving the pyrrolic N--H bonds of bacteriopheophytin, also are important in this process. PMID- 6275919 TI - Inactivation of voltage-gated delayed potassium current in molluscan neurons. A kinetic model. AB - Voltage-gated delayed potassium current in molluscan neurons is characterized by a marked inactivation. Inactivation can accumulate between repetitive pulses, giving rise to current patterns in which the maximum current during a second voltage pulse is less than the current at the end of the preceding pulse (cumulative inactivation). Other features of inactivation of this current include an onset time-course that can be characterized by the sum of two exponential processes and an early minimum in the recovery-vs.-time curve. A simple four state model is developed that can, when supplied with rate constants derived from voltage-clamp experiments, reproduce these features of inactivation. The model incorporates state-dependent inactivation rates. Upon depolarization, both open and closed channels can be inactivated, although inactivation of closed channels is much faster. Upon repolarization, recovery from inactivated states is sufficiently slow that little recovery occurs during a short interpulse interval. Cumulative inactivation comes about as a result of fast inactivation during the second pulse, further limiting the peak current from the level at the end of the previous pulse. PMID- 6275920 TI - Reaction of C-type cytochromes with the iron hexacyanides. Mechanistic implications. AB - The reaction of c-cytochromes with iron hexacyanides is similar in mechanism to the interaction of cytochromes with their physiological oxidants and reductants in that the formation of complexes precedes electron transfer. Analysis of the kinetics of oxidation and reduction of a number of c-cytochromes by solving the simultaneous differential equations defining the mechanism is possible, and allows assignment of all six rate constants describing a minimum three-step mechanism [cyto(Fe(+3)) + Fe(+2) right harpoon over left harpoon cyto (Fe(+3)) - Fe(+2) right harpoon over left harpoon cyto(Fe(+2)) - Fe(+3) right harpoon over left harpoon cyto(Fe(+2)) + Fe(+3)]. We find that the usual steady-state approximations are not valid. Furthermore, the ratio of first-order rate constants for electron transfer was approximately 1.0, and no correlation was found between any of the six rate constants and the differences in oxidation reduction potential of the iron-hexacyanides and different cytochromes c. However, it was found that the ratio of the rate constants for complex formation between ferricytochrome c and potassium ferrocyanide and ferrocytochrome c and potassium ferricyanide was proportional to the difference in oxidation-reduction potentials. Thus the minimum three-step mechanism given above accurately describes the observed kinetic data. However, this mechanism leads to a number of conceptual difficulties. Specifically, the mechanism requires that the collision complexes formed [cyto(Fe(+3)) - Fe(CN)(6) (-4) and cyto(Fe(+2)) - Fe(CN)(6) ( 3)] have very different equilibrium constants, and further requires that formation of the collision complexes be accompanied by "chemistry" to make the intermediates isoenergetic. A more complex five-step mechanism which requires that the reactants [Fe(CN)(6) (-4) and ferricytochrome c or Fe(CN)(6) (-3) and ferrocytochrome c] form a collision complex followed by a first-order process before electron transfer, was found to yield results similar to those of the three-step mechanism. However, describing the formation of the collision complex in terms of a rapid equilibrium circumvents conceptual difficulties and leads to a physically reasonable mechanism. In this mechanism the reactants are in rapid equilibrium with the collision complexes and the rate constants for complex formation are controlled by diffusion and accessibility. The collision complexes then rearrange, possibly through conformational changes and/or solvent reorganization, to yield isoenergetic intermediates that can undergo rapid reversible electron transfer. The five-step mechanism can be described by the same rate constants obtained from the three-step mechanism with the appropriate adjustments to account for rapid equilibrium. This more complex analysis associates the oxidation-reduction potential of a particular cytochrome with the relative magnitude of the first-order conversion of the oxidant and reductant collision complexes to their respective intermediates. Thus the cytochromes c control their oxidation-reduction potential by chemical and/or structural alterations. This mechanism appears to be general in that it is consistent with the observed kinetics of 11 different cytochromes c from a wide variety of sources with a range of oxidation-reduction potentials. PMID- 6275921 TI - Dynamics of potassium ion currents in squid axon membrane. A re-examination. AB - The original experiments of Cole and Moore (1960. Biophys. J. 1:161-202.), using conditioning and test membrane potentials to examine the dynamics of the potassium channel conductance in the squid axon, have been extended to test voltage levels by the use of tetrodotoxin to block the sodium conductance. The potassium currents for test voltage levels from -20 to +85 mV were superposable by translation along the time axis for all conditions tested: (a) with depolarizing conditioning voltages; (b) with hyperpolarizing conditioning voltages; and (c) in normal and in high potassium external media. The only deviations from superposition seen were when the internal sodium concentration was abnormally high and the potassium currents showed saturation at high levels of depolarization. Some restoration toward normal kinetics could be obtained by rapidly repeated depolarizations. PMID- 6275922 TI - Potassium ion currents in the crayfish giant axon. Dynamic characteristics. AB - The kinetics of the voltage-sensitive potassium channel in crayfish axon have been examined. The conductance increase after a step depolarization from rest can be described by a first-order kinetic process raised to the third power. When conditioning voltage levels preceded the test pulse, the steady-state conductance was found to be independent of initial conditions. Depolarizing conditioning voltages in general allowed superposition of test voltage potassium currents by a shift along the time axis. Hyperpolarizing conditioning voltages produced a delay in onset of conductance during the test pulse and changed the kinetics so that superposition was not possible. The delay increased during the hyperpolarization with a first-order lag having a time constant in the range of 1.5-3 ms. Return to the resting level caused recovery from the delayed state to follow a single exponential decay with a time constant of 1.9-2.2 ms. The steady state delay vs. voltage curves were not saturated at potentials as negative as -180 mV. PMID- 6275923 TI - Rotational dynamics of protein and boundary lipid in sarcoplasmic reticulum membrane. AB - We have used spin labels and electron paramagnetic resonance (EPR) to study the correlation between the rotational dynamics of protein and lipid in sarcoplasmic reticulum (SR) membranes. A short-chain maleimide spin label was used to monitor the submillisecond rotational mobility of the Ca-ATPase enzyme (using saturation transfer EPR); a free fatty acid spin label was used to monitor the submicrosecond rotational mobility of the bulk lipid hydrocarbon chains (using conventional EPR); and a fatty acid spin label derivative (long-chain maleimide) attached to the enzyme was used to monitor the mobility of hydrocarbon chains adjacent to the protein (i.e., boundary lipid). In the native SR membranes, the protein was highly mobile (effective correlation time 50 microseconds). The spectra of the hydrocarbon probes both contained at least two components. For the unattached probe, the major component indicated nearly as much mobility as in the absence of protein (effective rotational correlation time 3 ns), while a minor component, corresponding to 25-30% of the total signal, indicated strong immobilization (effective correlation time greater than or equal to 10 ns). For the attached hydrocarbon probe, the major component (approximately 70% of the total) was strongly immobilized, and the mobile component was less mobile than that of the unattached probe. When the lipid-to-protein ratio was reduced 55% by treatment with deoxycholate, protein mobility decreased considerably, suggesting protein aggregation. A concomitant increase was observed in the fraction of immobilized spin labels for both the free and attached hydrocarbon probes. The observed hydrocarbon immobilization probably arises in part from immobilization at the protein-lipid boundary, but protein-protein interactions that trap hydrocarbon chains may also contribute. When protein aggregation was induced by glutaraldehyde crosslinking, submillisecond protein mobility was eliminated, but there was no effect on either hydrocarbon probe. Thus protein aggregation does not necessarily cause hydrocarbon chain immobilization. PMID- 6275924 TI - ESR spin-label studies of lipid-protein interactions in membranes. AB - Lipid spin labels have been used to study lipid-protein interactions in bovine and frog rod outer segment disc membranes, in (Na+, K+)-ATPase membranes from shark rectal gland, and in yeast cytochrome oxidase-dimyristoyl phosphatidylcholine complexes. These systems all display a two component ESR spectrum from 14-doxyl lipid spin-labels. One component corresponds to the normal fluid bilayer lipids. The second component has a greater degree of motional restriction and arises from lipids interacting with the protein. For the phosphatidylcholine spin label there are effectively 55 +/- 5 lipids/200,000 dalton cytochrome oxidase, 58 +/- 4 mol lipid/265,000 dalton (Na+, K+)-ATPase, and 24 +/- 3 and 22 +/- 2 mol lipid/37,000 dalton rhodopsin for the bovine and frog preparations, respectively. These values correlate roughly with the intramembrane protein perimeter and scale with the square root of the molecular weight of the protein. For cytochrome oxidase the motionally restricted component bears a fixed stoichiometry to the protein at high lipid:protein ratios, and is reduced at low lipid:protein ratios to an extent which can be quantitatively accounted for by random protein-protein contacts. Experiments with spin labels of different headgroups indicate a marked selectivity of cytochrome oxidase and the (Na+, K+)-ATPase for stearic acid and for cardiolipin, relative to phosphatidylcholine. The motionally restricted component from the cardiolipin spin label is 80% greater than from the phosphatidylcholine spin label for cytochrome oxidase (at lipid:protein = 90.1), and 160% greater for the (Na+, K+) ATPase. The corresponding increases for the stearic acid label are 20% for cytochrome oxidase and 40% for (Na+, K+)-ATPase. The effective association constant for cardiolipin is approximately 4.5 times greater than for phosphatidylcholine, and that for stearic acid is 1.5 times greater, in both systems. Almost no specificity is found in the interaction of spin-labeled lipids (including cardiolipin) with rhodopsin in the rod outer segment disc membrane. The linewidths of the fluid spin-label component in bovine rod outer segment membranes are consistently higher than those in bilayers of the extracted membrane lipids and provide valuable information on the rate of exchange between the two lipid components, which is suggested to be in the range of 10(6)-10(7) s 1. PMID- 6275925 TI - The active form of cytochrome c oxidase: effects of detergent, the intact membrane, and radiation inactivation. AB - Cytochrome oxidase is a multisubunit, intrinsic membrane protein with a complex function that includes oxidation of cytochrome c, reduction of oxygen and generation of a membrane potential. To clarify the relationship of its normal function to protein and membrane structure, we have examined the kinetic behavior of rat liver cytochrome oxidase in the intact inner mitochondrial membrane and in detergent solubilized states. Dissolution of rat liver mitochondrial membranes alters the kinetic parameters of the oxidase in a manner dependent in part on the dispersing agent, and characterized by a large increase in maximal activity which is not attributable to exposure of more oxidase or diminished affinity for cytochrome c. The most profound effect of solubilization of the membrane is seen on the low affinity reaction of cytochrome c, suggesting that the electron transfer pathway from this site to oxygen is sensitive to alterations in hydrophobic interactions within the oxidase. Purified rat liver and beef heart oxidase exists predominantly in a monodisperse, 300 kilodalton form in laurylmaltoside (Rosevear et al., 1980). However, a smaller, 130 kd species that exhibits high turnover rates equal to the 300 kd form is detected in some beef heart preparations, implying that the dimer may not be essential for high activity. Radiation inactivation studies on purified oxidase reveal a molecular weight for the functional unit of approximately 70 kd. It is concluded that less than a complete set of subunits may be sufficient for both normal binding of cytochrome c and rapid electron transfer to oxygen. PMID- 6275927 TI - Melting experiment concerning the topological structure of closed circular double stranded DNA. PMID- 6275926 TI - High resolution nuclear magnetic resonance studies of the conformation and orientation of melittin bound to a lipid-water interface. AB - Previously, the size and stoichiometry of mixed micelles of perdeuterated dodecylphosphocholine and melittin were characterized and the 1H NMR spin systems of most amino acid residues of micelle-bound melittin identified. One- and two dimensional 1H-1H Overhauser experiments have now been used to obtain qualitative information on intramolecular proton-proton distances. These data show that the N terminal and the C-terminal segments of melittin form two spatially distinct, compact domains; using lipid spin labels these could be located near the micelle surface. For the C-terminal domain a detailed conformation was determined by using the distance contraints from the Overhauser studies as input for a distance geometry algorithm. PMID- 6275928 TI - The mass spectra of the trimethylsilyl derivatives of the hydroxy and acid metabolites of delta 1- and delta 6-tetrahydrocannabinol. AB - Deuterium labelling and high resolution mass measurements have been used to investigate the fragmentation mechanisms leading to diagnostic ions in the mass spectra of the trimethylsilyl derivatives of 58 hydroxy and acid metabolites of delta 1- and delta 6-tetrahydrocannabinol and of two related compounds, 2 alpha- and 2 beta-hydroxy-delta 6-tetrahydrocannabinol. The spectra of most of the hydroxy metabolites contained abundant ions which were characteristic of the position of hydroxy substitution. These could be used diagnostically to determine the structures of polysubstituted metabolites. PMID- 6275929 TI - [Use of the proton magnetic relaxation technic in the study of experimental burns]. AB - Experiments with the use of proton magnetic relaxation have shown the presence of marked biophysical changes in the organs during deep skin burns with an area of 6% of the body surface. The method is highly responsive to the changes in tissue hydratation and is characterized by good reproducibility of the results. The technical simplicity of the method (5-10 minutes per sample) permits using proton magnetic relaxation on a large scale. The data obtained allow a conclusion that the method under consideration should be recommended for studying the status of water in experimental burns and in clinical trials. PMID- 6275930 TI - ["Micro-lead" technic for recording ion currents through the membranes of individual myocardial cells]. AB - Patch-voltage-clamp method has been used for registration of the fast inward current in single heart muscle cells. Ionic current were measured on a small patch of membrane which is of about 1/200-1/300 of the total cell surface (diameter of patch is of about 5 micrometer). To correct the distortions of raw signal caused by the stray capacity in feedback of current-voltage converter and adjustable increasing of high-frequency gain was used. This allows one to shorten the net time of transient process in response to switching the command potential up to 30 microseconds. The channel population in membrance patch of micrometer in diameter is, on the one hand, large enough to record directly the integral ionic current and is, on the other hand, small enough for fluctuations of ionic current to be pronounced. This circumstance permits the method to be applied for ionic current investigations both by the classical methods and by statistical analysis. PMID- 6275931 TI - Hexose transport in Epstein-Barr virus (EBV) negative lymphoma lines and their EBV converted, virus genome carrying sublines. AB - Increased hexose uptake is a marker for viral transformation, as has been shown in non-human fibroblasts transformed by oncogenic viruses. If this phenomenon is a general expression of viral induced transformation it should also apply on different oncogenic virus-cell systems. Recently two human EBV-negative lymphoma lines were converted to a stable EBV-positive state by infection with EBV. According to their biochemical and biological properties they enable us to study events associated with EBV-transformation. We analysed the uptake of (3H) glucosamine and (3H) 2-deoxy-D-glucose into BJAB and Ramos and their EBV converted sublines and found a clear increase of the rate of uptake of both sugars in the EBV-positive sublines. Control experiments confirmed that the increased uptake was due to alterations on the level of the hexose membrane carriers and not due to increased metabolism. The observation of increased hexose uptake in the only presented available virus transformed human cell system is a strong argument for the general importance of this transformation-associated membrane change. PMID- 6275932 TI - Cardiovascular actions and interaction of chlordimeform in the dog. PMID- 6275933 TI - Etiology of childhood diarrhea and oral rehydration therapy in northeastern Brazil. PMID- 6275934 TI - Postexposure immunoprophylaxis against B virus (Herpesvirus simiae) infection. AB - Local infiltration of antiserum into sites inoculated with B virus protected rabbits from an otherwise fatal encephalomyelitis. Treatment was effective when delayed for six hours but not after 24 hours. Homologous rabbit antisera were more effective than heterologous monkey antisera, and protection was unrelated to neutralisation titres. Protection apparently depended not on neutralisation of inoculated virus but on destruction of infected cells before they produced progeny virus. Normal human immunoglobulin able to neutralise B virus did not protect. Intravenously administered antibody was effective only if large doses were given. The findings suggest that persons bitten or scratched by monkeys latently infected with B virus may be treated successfully by immunoprophylaxis with specific antibody. Stocks of human or of more readily available simian antisera should be held in laboratories where such animals are used. PMID- 6275935 TI - Is bran useful in diverticular disease? PMID- 6275936 TI - Doubts about diverticular disease. PMID- 6275937 TI - Physiological changes underlying jet lag. PMID- 6275938 TI - Immune response of infants in tropics to injectable polio vaccine. PMID- 6275939 TI - Extensor digitorum brevis--a predictor of neuropathy in the leg? PMID- 6275941 TI - Confirmation of herpes simplex virus type 2 infections in herpes-like genital lesions by a simple complement-fixation test. AB - The presence of complement-fixing antibody to an early herpes simplex virus type 2 (HSV-2) antigen (the AG-4 antigen) was correlated with HSV-2 infection in the sera of patients with genital herpes. Eighty-eight per cent of sera taken two weeks after clinical diagnosis of a primary or recurrent herpes infection in patients, confirmed to have HSV-2 by virus isolation and typing, contained the anti-AG-4 complement-fixing antibody. None of the patients with genital HSV-1 had the antibody, and only 9% of controls or patients with facial HSV-1 infection had positive results for the antibody. This correlation was used to identify genital HSV-2 infections when either no virus sample had been taken or when virus isolations had been unsuccessful. Thus, a simple complement-fixation test can confirm an HSV-2 virus infection without isolation of the virus from the herpetic lesion. PMID- 6275940 TI - Incidence of herpes simplex virus types 1 and 2 isolated in patients with herpes genitalis in Sheffield. AB - Thirty-one strains of herpesvirus (HSV), isolated from patients presenting with the clinical features of herpes genitalis, were typed by polypeptide analysis of virus proteins in sodium dodecyl sulphate polyacrylamide gels. Nineteen (61.3%) of the isolates were shown to be HSV type 1 and 12 (38.7%) HSV type 2. There was no obvious difference in the incidence of HSV-1 in primary or recurrent infections and no apparent correlation between the genital site of isolation and virus type. The high incidence of genital HSV-1 infection in this group of patients is probably due to the increased practice of oro-genital contact and has possible implications for the future development of drugs and vaccines in the control of genital herpes. PMID- 6275942 TI - Facilitation of monosynaptic reflexes by voluntary contraction of muscle in remote parts of the body. Mechanisms involved in the Jendrassik Manoeuvre. AB - The facilitation of the tendon reflex of quadriceps, induced by voluntary contraction of a muscle group in the upper limb, evolves in the three phases and depends on several factos. These include the strength and type (ballistic or ramp) of the contraction. In the absence of any electromyographic activity in the conditioning muscle (from a lesion of its nerve trunk or by blockage of conduction by injection of xylocaine), a voluntary attempt to contract it causes only a moderate facilitation of the quadriceps motor nucleus; while reflex activation of the conditioning muscles, by vibration or sudden stretching. causes a more marked facilitation. Thus, it is concluded that at least two distinct mechanisms act sequentially. First, a general motor facilitation of supraspinal origin induces the first moderate phase of facilitations (phase I). This early facilitation precedes the onset of electromyographic activity in the conditioning muscle. Secondly, the more marked facilitation (phase II) is related to the stimulation of afferents originating from the conditioning muscles, either by fusimotor excitation in voluntary contraction or by manoeuvres which stimulate the spindle receptors mechanically. To investigate the transmission pathways of these facilitations, the latencies of these two phases were compared at two distant motor nuclei, the trigeminal motor nucleus and the quadriceps nucleus, following voluntary contractions of the tibialis anterior. Since the facilitations of the trigeminal motor nucleus precede those of the quadriceps motor nucleus, a slowly-travelling rostro-caudal facilitation is postulated during phase II when the facilitation is related to afferents coming from the contracting muscle. A long loop, therefore, is postulated to interpret the second phase of the facilitation curves. PMID- 6275943 TI - Neostriatal enkephalin-immunoreactive neurones project to the globus pallidus. AB - The origin of enkephalin-immunoreactive nerve terminals in the globus pallidus was investigated by combining immunocytochemistry with stereotaxic injection of neurotoxic agents (colchicine and kainic acid) and microknife deafferentations. The intracerebral administration of colchicine, irrespective of whether in the caudate putamen or in globus pallidus, induces the appearance of enkephalin immunoreactive cell bodies and fibres in the caudate putamen. No immunoreactive cell bodies were depicted in the globus pallidus after this treatment. Kainic acid injections in the caudate putamen produced topographic depletions of enkephalin-immunoreactive terminals in the globus pallidus. The more anterior injections produced medial-anterior depletions, while posterior injections gave latero-posterior depletions. Injections in the globus pallidus produced only a non-specific loss of fluorescence restricted to the tip of the cannula. Coronal microknife cuts produced a combination of build-up and depletion of enkephalin immunofluorescence according to the position of the cut. The build-up of immunoreactive materials was always observed in the caudate-putamen side of the cut while depletions observed in the globus pallidus were related to the extent of deafferentation of this nucleus from the caudate putamen. All these observations confirmed the neostriatal origin (caudate putamen) of the enkephalinergic fibres present in the paleostriatum (globus pallidus). PMID- 6275944 TI - ACTH 1-24 and lysine vasopressin selectively activate dopamine synthesis in frontal cortex. AB - The accumulation of [3H] catecholamines from [3H] tyrosine in frontal cortical, septal, striatal and hippocampal slices was examined following intracerebroventricular (i.c.v.) injections of ACTH 1-24, lysine vasopressin (LVP) and saline. Both ACTH 1-24 and LVP (1 microgram) selectively increased the accumulation of [3H] dopamine (DA) in frontal cortical slices, but did not affect that of [3H] norepinephrine (NE). LVP but not ACTH 1-24 also inhibited the accumulation of [3H] DA in striatal slices. ACTH 1-24 did not alter the accumulation of [3H] NE in hippocampal slices, nor did LVP alter the accumulation of either catecholamine (CA) in septal slices. In vitro incubations with ACTH analogs of LVP failed to alter the rate of accumulation of [3H] CAs in striatal, substantia nigral and frontal cortical slices, except for an inhibitory effect at high doses. This effect is believed to be an artifact of precursor dilution caused by release of tyrosine following degradation of the peptides. Neither peptide modified the increased [3H] CA accumulation stimulated by 26 mM K+, nor did ACTH 1-24 modify the inhibition of [3H] CA accumulation caused by 3 X 10 -6 M haloperidol or 3 X 10 -7 M apomorphine. Selective activation of the mesocortical DA system has also been reported ot occur in response to footshock, suggesting the possibility that endogenous ACTH and/or LVP might mediate the stress-induced activation of mesocortical DA synthesis. Alternatively, i.c.v. injections of these peptides may themselves be stressful and thus indirectly elicit the response. PMID- 6275945 TI - Lesions in nucleus reticularis gigantocellularis: effect on the antinociception produced by micro-injection of morphine and focal electrical stimulation in the periaqueductal gray matter. AB - The effect of bilateral electrolytic lesions in the nucleus reticularis giganto cellularis (NGC) on the antinociceptive efficacy of morphine and electrical stimulation applied in the periaqueductal central gray matter (PAG) was investigated. Antinociception, evaluated by standard hot plate and tail-flick analgesiometric tests, was reliably produced by morphine (5 microgram) and focal electrical stimulation (40-200 micro A) administered in the PAG of rats via chronic indwelling cannula/electrode assemblies. Subsequent to the initial antinociceptive testing, bilateral electrolytic lesions were introduced in the NGC and the antinociceptive efficacy of morphine and stimulation in the PAG was again evaluated. Lesions in the NGC prevented the expression of the antinociception produced by the microinjection of morphine in the PAG whereas the antinociception resulting from electrical stimulation in the PAG was unaffected. Further, lesions in the NGC did not alter baseline (control) nociceptive thresholds in either analgesiometric test. These results provide additional support for involvement of the NGC in morphine-induced antinociception and, in addition, suggest that the NGC is not essential to a tonically-active inhibitory system or to the antinociception produced by focal electrical stimulation in the PAG. PMID- 6275946 TI - Evidence for GABA receptors on serotonergic afferent terminals in the substantia nigra and on nigral efferent projections to the caudal mesencephalon. AB - Changes in [3H]GABA binding in rat substantia nigra (SN) were examined after (1) hemitransections at various levels posterior to SN and (2) 5, 7 dihydroxytryptamine (5, 7-DHT) induced lesions of the dorsal or median raphe or SN. Hemitransections 2 mm caudal to SN resulted in a 52% decrease in specific high affinity [3H] GABA binding in SN when measured 2-3 weeks postoperatively. No significant change in nigral GABA binding was found in rats with hemitransections placed at a postcollicular level(through the rostral pons). 5, 7-DHT placements in median raphe, or directly into SN, caused a 27% decrease in specific [3H] GABA binding in SN. 5, 7-DHT lesions of the dorsal raphe did not significantly affect the binding of [3H]GABA in SN. These data suggest that a significant protion of GABA receptors in SN may be located presynaptically on axon terminals of serotonergic projections from the median raphe to SN. Other pathways between the SN and the caudal mesencephalon also appear to contain GABA receptors; these are likely to be efferent nigral projections to the mesencephalic reticular formation. PMID- 6275947 TI - Analgesic dipeptide, kyotorphin (Tyr-Arg), is highly concentrated in the synaptosomal fraction of the rat brain. AB - To examine the physiological role of the analgesic dipeptide, kyotorphin (Tyr Arg), which was isolated from the mammalian brain, its subcellular localization was studied. Kyotorphin was determined using high-performance liquid chromatography (HPLC) with an electrochemical detector. This dipeptide was found to be concentrated in the crude mitochondrial (P2) fraction. Further subfractionation of the P2 fraction revealed that kyotorphin was exclusively localized in the synaptosomal fraction.This finding suggests the possibility that kyotorphin has a neurotransmitter/neuromodulator role in the brain. PMID- 6275948 TI - Morphine-induced hyperthermia: lack of cross-tolerance with enkephalin analogs. PMID- 6275949 TI - Prolonged depression of pelvic ganglion transmission -- a peripheral manifestation of spinal cord transection. PMID- 6275950 TI - Deoxyglucose uptake in the rat thoracolumbar cord during activation of aortic baroreceptor afferent fibers. PMID- 6275951 TI - Hormonal influences on seizure kindling: the effects of post-stimulation ACTH or cortisone injections. AB - Repeated application of brain stimulation can lead to a progressively augmenting electrical and behavioral response-- a phenomenon termed seizure kindling. In this experiment, stimulation was delivered once per day, and was followed by peripheral (intraperitoneal) administration of ACTH or cortisone. An intermediate or a high dose of either hormone (0.3 IU or 3.0 IU of ACTH/animal, 10 mg or 25 mg cortisone/animal) delayed the completion of kindling if administered shortly after each kindling stimulation. Lower doses (0.03 IU of ACTH or 2 mg of cortisone) had no significant effects. The high dose of ACTH or cortisone was no longer effective if administration was delayed more than 4 h after stimulation. Peripherally administered ACTH and cortisone can influence processes initiated by the brain stimulation which presumably underlie the augmentation of response to successive stimulations. This time-limited action is analogous to the effects of these hormones on memory consolidation. PMID- 6275952 TI - Beta-endorphin concentrations in serum, hypothalamus and central gray of hypophysectomized and mediobasal hypothalamus lesioned rats. AB - The serum and brain concentrations of beta-endorphin immunoreactivity have been studied in intact, mediobasal hypothalamus (MBH) lesioned and hypophysectomized male rats. After hypophysectomy there is a major reduction (90%) of beta endorphin concentration in the serum but only a partial reduction (20%) in the mediobasal hypothalamus. However, MBH lesions enhance beta-endorphin serum values in previously hypophysectomized rats. Long-term MBH lesions alone lead to an almost complete disappearance of beta-endorphin in the central gray matter with a slight decrease in the serum. These data clearly show that: (1) the pituitary is the major source of beta-endorphin in the serum; (2) the hypothalamus is the major source of beta-endorphin in the central gray matter; and (3) there is clear influence of the pituitary on hypothalamic beta-endorphin. PMID- 6275953 TI - Altered neuronal responsiveness to biogenic amines in rat cerebral cortex after serotonin denervation or depletion. AB - To further investigate monoaminergic mechanisms in cerebral cortex, responsiveness of cortical neurons to microiontophoretic applications of serotonin (5-HT), dopamine (DA) or noradrenaline (NA) was examined in the frontoparietal region of control, 5,7-dihydroxytryptamine (5,7-DHT)- and p chlorophenylalanine (PCPA)-treated rats anesthetized with urethane. As a rule, 100 nA applications of either one of these biogenic amines induced marked slowing or total interruption of 'spontaneous' firing overlasting the 30 s period of ejection. Given the large amounts of monoamines ejected, it could be inferred that such microiontophoretic applications produced a maximal activation of receptors. In control rats, the responses to 5-HT, DA and NA were of approximately equal duration (approximately equal to 5 min). Two to 4 weeks after denervation with 5,7-DHT, most neurons (75%) exhibited greatly prolonged responses to 5-HT (approximately equal to 14 min), and marked depressions of firing could be induced by small ejection currents (approximately equal to 2 nA) having little or no effect in the controls. In addition, 85% of the units supersensitive to 5-HT showed considerably shortened responses to DA and NA (approximately equal to 1 min). After 2-14 days of depletion with PCPA, there was no change in the responsiveness to 5-HT in spite of a 91% lowering of cortical 5 HT content equivalent to that measured after denervation. Nevertheless, responsiveness to DA and NA was again diminished in a majority (80%) of the units tested. In control or PCPA-treated rats, acute administration of the 5-HT re uptake blocker fluoxetine increased the duration of depressions induced by 100 nA applications of 5-HT but did not enhance responsiveness to low ejection currents. This suggested that, after 5-HT denervation, the suppression of re-uptake was mainly responsible for the prolongation of 5-HT responses ('presynaptic' component of supersensitivity), whereas a modification of 5-HT receptors accounted for the greater efficacy of small doses of 5-HT ('postsynaptic' component). Responsiveness to the microiontophoretic application of phenylephrine (PHE), a noradrenergic a-agonist, was comparable with that to NA in PCPA- and 5,7 DHT-treated as well as in control rats. Therefore, the hyposensitivity to DA and NA appeared indicative of a desensitization of catecholamine receptors caused by the absence of 5-HT. Such a desensitization may be viewed as an adaptive change resulting from an increased release of endogenous DA and NA. This interpretation would in turn imply that, normally, 5-HT regulates catecholamine release in the neocortex. PMID- 6275954 TI - Periodic fluctations in synaptic transmission and enhancement of transmission in hippocampal slices. AB - The properties of the synchronously activated radiatum fiber-CA1 synaptic population were examined with the in vitro hippocampal slice preparation. Periodic fluctuations in synaptic transmission and in the enhancement of synaptic transmission were observed with periods ranging from 8 to 20 s. Such periodic fluctuations did not arise from fluctuations in afferent radiatum fiber activity. The period and amplitude of the cyclic variations in the enhancement of synaptic transmission were found to be altered with repeated electrical stimulation of the radiatum fibers. These results reflect cooperative synaptic actions which must be taken into consideration in the delineation of the mechanisms of potentiation. PMID- 6275956 TI - Reduction of polyneuronal innervation of muscle cells in tissue culture after long-term indirect stimulation. PMID- 6275955 TI - Immunocytochemical localization of beta-endorphin (lipotropin C-fragment) in the developing rat spinal cord and hypothalamus. AB - Immunocytochemical studies have been performed on rat spinal cord and hypothalamus during development, using an antibody to beta-endorphin. Specific immunoreactivity was demonstrated in histological sections of spinal cord, in ventral and dorsal horn cells and nerve fibres, in the meningeal layer, the ependymal lining of the central canal, and in the endothelium of the ventral spinal artery and other blood vessels. beta-Endorphin immunoreactivity was also distributed widely in neurones, central and peripheral nerve fibres, and in non neuronal cells in cultures of spinal cord tissue explanted from rat embryos 7--10 days before birth. Immunofluorescence disappeared abruptly after the 28th postnatal day in vivo, and in the fourth week of incubation of embryonic spinal cultures. In contrast, cultures of the ventral diencephalic (hypothalamic) region of embryonic brain at the same gestational age showed a characteristically different pattern of beta-endorphin immunoreactivity which persisted for more than 7 weeks. The results provide evidence for the biosynthesis of a beta endorphin-like peptide in rat spinal cord during development. The biosynthesis terminates at an early stage both in vivo and in vitro, suggesting that control of the biosynthesis is intrinsic to spinal cord tissue and possibly to the peptide-producing cells themselves. PMID- 6275957 TI - Development of the rat septohippocampal projection: a retrograde fluorescent tracer study. PMID- 6275958 TI - Development of beta-adrenergic receptors and the in vitro accumulation of cyclic AMP in the chick spinal cord. AB - To clarify the functional development of the descending monoaminergic input to the chick spinal cord we have studied the ontogeny of beta-adrenergic receptors by measuring the specific binding the tritiated dihydroalprenolol (DHA). In addition, we examined the ability of isoproterenol to stimulate the accumulation of cyclic AMP in slices of developing chick spinal cord. Results show that the chick spinal cord contains a high density of beta-adrenergic receptors that are apparently linked to adenylate cyclase. During development, both the density of beta-receptors, as determined by the specific binding of DHA, and the response of tissue slices to isoproterenol underwent marked changes. beta-Adrenergic receptors (approximately 4 fmol/mg tissue) were first detected on the fourteenth day in ovo. Receptor density increased to approximately 20 fmol/mg by day 20. Between day 20 and the time of hatching, a sharp increase in receptor density, to approximately 50 fmol/mg, was seen. The density of receptors remained high until the second day after hatching, fell off to approximately 30 fmol/mg by the fourth day, and remained relatively unchanged through day 30. The response of spinal cord slices to isoproterenol showed a similar pattern of development with the peak response (7-fold increase in levels of cyclic AMP) occurring at or near the time of hatching. During the period between day 18 in ovo and the time of hatching, when both the response of tissue slices to isoproterenol and the density of beta-receptors increased markedly, the activity of phosphodiesterase did not change. Therefore, the pronounced changes in adrenergic responsiveness that occurred near the time of hatching appear to be related primarily to changes in the density of beta-adrenergic receptors coupled to adenylate cyclase. Such developmental changes in the density of beta-adrenergic receptors and adrenergic responsiveness may play an important role in determining the functional state of the descending monoaminergic systems in the chick spinal cord. PMID- 6275959 TI - Ontogenesis of GABA receptor sites in chick embryo cerebellum. AB - The time-course of the development of GABA receptor sites in chick embryo cerebellum was correlated with the appearance of synaptic junctions in the cerebellar cortex. At 13 days of incubation, the earliest stage examined, specific [3H]GABA binding was only 19% of that found in cerebella of adult chicks. Between 15 days of incubation and hatching, specific [3H]GABA binding increased 3-fold, already reaching at birth adult values. During this period the number of synaptic junctions also increased. Scatchard analysis of the binding data obtained at birth revealed two binding sites of KdS 40 and 174 nM and a maximal number of binding sites (n) of about 1.6 and 4.0 pmol/mg protein, respectively. The high-affinity binding site for [3H]GABA was inhibited by muscimol, GABA, imidazoleacetic acid and bicuculline (IC50: 0.007, 0.020, 0.1 and 10 microM, respectively). These values correspond to the potencies shown by those compounds in the binding to the synaptic GABA receptor. Treatment of the synaptic membranes with Triton X-100 enhanced [3H]GABA binding depending on the developmental stage studied, suggesting that GABA-modulin that inhibits the binding also appears during that period. PMID- 6275960 TI - Brain beta-endorphin-like immunoreactivity in adult rats given beta-endorphin neonatally. AB - beta-endorphin-like immunoreactivity was measured by radioimmunoassay in the brains of adult rats treated neonatally with beta-endorphin, naloxone, or vehicle. After treatment with beta-endorphin, the decreases observed in beta endorphin-like immunoreactivity in the hypothalamus, pineal, midbrain, pons medulla, hippocampus, striatum, frontal cortex, occipital cortex, and posterior cortex were highly significant but the 23% decrease in the thalamus was not significantly different from that of control rats. Neonatal administration of naloxone only resulted in a significant decrease in beta-endorphin-like immunoreactivity in the hypothalamus. In contrast, no differences were discernible in content of either beta-endorphin-like immunoreactivity or ACTH like immunoreactivity in the pituitary of rats treated with beta-endorphin, naloxone, or vehicle in the neonatal period. These same rats had shown an increased threshold to painful thermal stimulation by the tail-flick test after administration of either beta-endorphin or naloxone at birth. The results suggest that neonatally injected beta-endorphin may alter the levels of beta-endorphin like immunoreactivity in rat brain as well as the response to pain. PMID- 6275961 TI - Delta opiate receptors are involved in the endopioid-induced myoclonic contractions. AB - Morphine, the prototype mu opiate receptor agonist, decreased the spontaneous and [D-Ala2]-Met-enkephalinamide (DALA)-induced myoclonic contractions (MC) of submandibular muscles in the anaesthetized rat. The proposed kappa receptor agonists ketocyclazocine, ethylketocyclazocine and bremazocine failed to induce MC. In addition, bremazocine inhibited the spontaneous and DALA induced MC. Cyclazocine, the so-called sigma opiate receptor agonist, had a weak potency in generation of MC, but without step dose response tendency. The most potent opioid peptide in inducing the MC and electrocortical (ECoG) epileptic pattern was the delta opiate receptor agonist [D-Ala2,D-Leu5]-enkephalin (DADL). All drugs were administered intraventricularly. The results indicate that myoclonic phenomena induced by DADL and probably by other endopioids are mediated by delta opiate receptors in the rat brain. It is suggested that the combined ECoG and EMG method used in this study offers an opportunity to define further the biological role of opiate receptors and to identify the potential delta opiate receptor acting drugs, which might provide a new approach to the therapy of some seizure disorders. PMID- 6275962 TI - [Epidemiologic, pathologic and clinical studies of 122 cancers of the stomach observed during 30 years in the CHU of Abidjan]. PMID- 6275964 TI - [Insensitivity of some rats to intraperitoneal injections of collagenase and trypsin]. AB - In the rat peritoneal injections of collagenase or trypsin give rise to severe lesions. In our experience 20% of the animals remain intact. The frequency of lesions increases with older and heavier subjects. Moreover 25% of the rats who remained free of lesions after a first injection of collagenase resist to a second one. This shows that they are strongly protected against the enzyme. The exact nature and location of this protective mechanism are not known. PMID- 6275963 TI - [Effects of the destruction of the suprachiasmatic nuclei on the circadian rhythms of ACTH corticosterone and the general activity of female rats exposed to a aperiodic environment]. AB - The effects of bilateral destruction of suprachiasmatic nuclei (SCN) on the temporal patterns of plasma concentrations in ACTH and corticosterone (sequential blood samples from an aortic cannula, at 4 h intervals over 48 h) and of general activity (stabilimeter recordings) were studied in female rats under constant illumination (LL : 10 lx). Under this arrhythmic environment, the SCN lesions induced a syndrome similar to that previously described in a photoperiodic environment (12 L-12 D). This SCN syndrome included : 1) blockade of the ACTH rhythm at baseline levels ; 2) maintained fluctuations of corticosterone, with either circadian or ultradian profiles ; 3) persistent rhythmicity of general activity, with a circadian periodicity in addition to ultradian periods. The persistence of a circadian rhythmicity in the SCN syndrome under arrhythmic environmental conditions, clearly argues in favour of the occurrence of endogenous components of the circadian pacemaker outside the SCN. PMID- 6275965 TI - Why patients seek unproven cancer remedies: a psychological perspective. AB - When it is discovered that cancer has recurred, patients experience a period of acute emotional distress, with increased anxiety, fear, helplessness, and depression. Well-meaning colleagues, friends, and relatives feel compelled to tell them about unorthodox cancer "cures." These unproven remedies tend to fit holistic health concepts purporting to enhance the body's own defenses; most involve the use of chemicals or drugs, nutritional supports, vitamins, vaccines, and mind-body techniques. In the past decade, laetrile has been the most popular unorthodox remedy. The distraught patient and family are deluged with information and have trouble evaluating what is valid. Physicians who treat patients with cancer should make sure that they themselves understand the emotional basis for a patient's need to pursue unorthodox remedies. The risk is heightened when the patient senses the doctor has "given up" and has "nothing more to offer." Patients should feel able to ask questions about unproven remedies without fear that the physician will be judgmental or punitive. Participation in a clinical trial of a new and promising treatment under investigation, within the full protection of ethical guidelines, should be suggested as an alternative to unproven treatments outside the medical system that are not subject to the same constraints. PMID- 6275966 TI - Dissolution of calcium hydroxyapatite. PMID- 6275967 TI - Factors in response to treatment of early postmenopausal bone loss. PMID- 6275968 TI - Development of the cyclic AMP response to parathyroid hormone and prostaglandin E2 in the embryonic chick limb. AB - The developing chick limb was studied to determine the ability of parathyroid hormone (PTH) and prostaglandin E2 (PGE2) to increase intracellular cyclic AMP (cAMP) during various stages of development. All developmental stages examined (stages 20-21, 24-25, and 26-28) responded to PGE2 when the cells were assayed immediately following the removal of the limbs from the embryos. In contrast, only stage 26-28 limb cells responded to PTH when assayed in a similar manner. The response to PTH was temporally correlated with the appearance of cartilage matrix in vivo. Undifferentiated limb cells were also cultured and assayed at various times for hormone responsiveness. Stage 24-25 high-density cell cultures responded initially to PGE2 but not to PTH. However, by 36 h and in all subsequent time intervals tested, the response to PTH was significantly greater than that to PGE2. The PTH receptor, in contrast to that of PGE2, was shown to be sensitive to trypsin treatment, but could be generated during subsequent cell culture. The majority of the hormone-responsive cells were found in cartilaginous regions of the limb, and were shown to respond to both hormones in a dose dependent manner. The PTH-induced cAMP response was affected by low cell density and mouse serum, both of which significantly inhibit the chondrogenic potential of cultured limb cells. These findings are consistent with a temporal correlation between the development of the PTH response and chondrogenesis in vivo. PMID- 6275970 TI - 1,25-dihydroxyvitamin D3 and the regulation of macrophage function. AB - Vitamin D3 deficient (D-) mice show a depressed inflammatory response and both inflammatory peritoneal macrophages and bone marrow polymorphonuclear leukocytes of D- mice exhibit a decreased spontaneous migration under agarose. The impaired phagocytic response of peritoneal macrophages from D- mice can be corrected by incubation with 1,25-dihydroxyvitamin D3 and is not affected by interaction with other vitamin D3 metabolites. Transfer of mice from the D- to the D+ state results in correction of both the inflammatory and the phagocytic response. Intactness of phagocyte function is thus directly dependent on vitamin D3 metabolism. PMID- 6275969 TI - Comparison of 1,25-, 25-, and 24,25-hydroxylated vitamin D3 binding in fetal rat calvariae and osteogenic sarcoma cells. PMID- 6275971 TI - Epidural beta-endorphin in treatment of pain. AB - Epidural administration of 3 mg of synthetic beta-endorphin produced analgesia in 10 patients with intractable pain due to disseminated cancer. Mean onset of relief of pain was 24 +/- 3 minutes and the mean duration of analgesia was 19 +/- 3 hours. The onset of analgesia produced by the epidural injection of beta endorphin was slower and the duration less than those observed after intrathecal injection. PMID- 6275973 TI - Myocardial glycolytic and gluconeogenic enzyme activity during cardiopulmonary bypass in humans. AB - Through a study of the activity of the regulatory enzyme of glycolysis phosphofructokinase and the regulatory enzyme of gluconeogenesis fructose-1,6 diphosphatase, myocardial metabolism has been evaluated in the human heart during cardiopulmonary bypass. Three ventricular transmural biopsy specimens were obtained at open-heart surgery before cardiopulmonary bypass was instituted and three similar specimens were taken 30 minutes after cardiopulmonary bypass was started. The enzyme assays were performed on the supernatant obtained from myocardial cells disrupted by sonication. The activity of both enzymes increased during the anoxic conditions of cardiopulmonary arrest. The increase in phosphofructokinase activity is likely due to an overall increase in glycolytic activity, but the presence and increased activity of fructose-1,6-diphosphatase was unexpected. PMID- 6275972 TI - Effect of enflurane and fentanyl on the clinical characteristics of long-term succinylcholine infusion. AB - The characteristics of the neuromuscular block produced by prolonged succinylcholine infusion were compared in 40 patients anaesthetized with either nitrous oxide with enflurane (1-2 per cent inspired) or nitrous oxide and fentanyl. Neuromuscular transmission was monitored using train-of-four stimulation and the infusion rate was adjusted to keep the first twitch at 10-15 per cent of its control value. Initially, all patients, exhibited a depolarizing type block all twitches of the train-of-four being roughly the same size, and the infusion rates were similar in the enflurane (54 microgram X kg-1/min) and the fentanyl (58 microgram X kg-1/min) groups. Tachyphylaxis developed later in both groups and correlated well with the onset of phase II block (dual block). This occurred sooner and at a lower cumulative dose in the enflurane group. Fourth to first twitch ratios decreased to 50, 25 and 0 per cent in 31, 46 and 59 minutes in the enflurane group, at cumulative succinylcholine doses of 2.2, 3.2 and 4.2 mg X kg-1 respectively. Corresponding figures for the fentanyl group were 52, 73 and 86 minutes, with dose of 3.4, 5.0 and 5.9 mg X kg-1. Infusion rates increased markedly after establishment of dual block, but were similar with enflurane (0.99 mg X kg-1/min) and fentanyl (1.12 mg X kg-1/min). Ten minutes after stopping the infusion fourth to first twitch ratios failed to reach 50 per cent in most patients given enflurane who had received more than 6 mg X kg-1 succinylcholine over more than 90 minutes. Corresponding figures for fentanyl patients were 13 mg x kg-1 and 150 minutes. The block in all 15 patients (9 enflurane, 6 fentanyl) who did not recover spontaneously was successfully antagonized with atropine and neostigmine. PMID- 6275974 TI - Mediastinal lymph-node biopsy is a definitive staging procedure for bronchogenic carcinoma. AB - Mediastinal lymph-node biopsy has been performed as a staging procedure in most cases of bronchogenic carcinoma since 1965. This study was carried out to assess the results of staging by this method in 75 patients. Biopsy sites were marked with hemoclips. Three zones were defined: zone 3 (inoperable), beyond the midline and above the lower border of the aortic arch; zone 1 (operable), more than 3 cm from the carcinoma, ipsilateral to the tumour; and zone 2, between zones 1 and 3. Forty of 75 patients had biopsy specimens showing carcinoma. The proportion was highest in small cell anaplastic carcinoma and lowest in squamous cell tumours. Of the 40 positive biopsies 35 were from zone 3. All the positive biopsy specimens showing small cell anaplastic carcinoma were from zone 3. Positive specimens correlated strongly with other evidence of inoperability. Mediastinal lymph-node biopsy is a definitive staging procedure. PMID- 6275975 TI - Causes of atypical pneumonia: results of a 1-year prospective study. AB - In a protocol study of cases of atypical pneumonia over a 1-year period an etiologic agent was established in 16 cases: Legionella pneumophila in 8, Coxiella burnetii in 3, Chlamydia trachomatis in 2, Mycoplasma pneumoniae in 1, para-influenza 3 virus in 1 and cytomegalovirus in 1. In the remaining 11 cases no agent was identified; the illnesses in these cases tended to be less severe. The pneumonia took much longer to resolve in the patients with Legionnaires' disease than in all the other patients (mean interval from onset of symptoms to clearing of the chest roentgenogram: 69 days v. an average of 16 days). However, the length of stay in hospital was similar for the three groups: those with Legionnaires' disease, those with atypical pneumonia of unknown cause and those with atypical pneumonia of various other established causes. L. pneumophila infection may explain a proportion of atypical pneumonias that previously could not be diagnosed, although in this series the cause of 41% of the pneumonias remained unexplained. PMID- 6275976 TI - The effect of chronic habitual alcohol intake on the development of liver cirrhosis and hepatocellular carcinoma: relation to hepatitis B surface antigen carriage. AB - To study the effects of habitual alcohol intake on the latency period for the development of liver cirrhosis and hepatocellular carcinoma (HCC), 158 patients with cirrhosis and 79 with HCC were analyzed with respect to age at the time of diagnosis. They were classified into four groups based on hepatitis B surface antigen (HBsAg) in serum, and the history of intake of more than one small bottle of Japanese "sake" or an equivalent per day for more than 10 years. The average age of HBsAg positive male cirrhotics with a drinking habit (n = 10) was 38.8 years, 10.5 years younger than that of those without a drinking habit (n = 8) (P less than 0.05). The average age of HBsAg negative cirrhotics with a drinking habit (n = 97) was 47.9 years, eight years younger than that of those not drinking (n = 36) (P less than 0.001). There was no significant difference in the laboratory data between these groups. The average age of the HBsAg positive HCC patients with a drinking habit (n = 20) was 48.9 years, nine years younger than that of those without a drinking habit (n = 12) (P less than 0.05). The average age of HBsAg negative male HCC cases with habitual intake of more than 126 ml of ethanol per day was 51.0 years (n = 8), ten years younger than that of nondrinking male HCC cases (n = 11) (P less than 0.05). These data suggest that habitual alcohol intake may promote the development of liver cirrhosis and HCC, especially in HBsAg carriers. PMID- 6275977 TI - Hepatitis B antigen in the liver in hepatocellular carcinoma in Shikoku, Japan. AB - An analysis of 105 autopsy cases (77 male, 28 female) of hepatocellular carcinoma (HCC) showed that 90 cases (85.7%) were associated with liver cirrhosis, of which 75 cases (83.3%) were of the macronodular type. Hepatitis B surface antigen (HBsAg) was detected with orcein stain in the hepatic tissue in 58 cases (55.2%). The incidence of HBsAg in cases of HCC patients with cirrhosis (58.9%) was higher, but not significantly, than that in those without cirrhosis (33.3%). The mean age of HBsAg-positive cases was five years less than that of HBsAg-negative cases, the age distributions therefore being significantly different. PMID- 6275978 TI - Intraductal carcinoma of the breast: follow-up after biopsy only. AB - Twenty-eight women with ductal carcinoma in situ (DCIS) of the breast treated by biopsy only were identified in a histologic review of 11,760 biopsies performed between 1950 and 1968. Seven of the 25 women followed for more than three years developed invasive breast carcinoma, all in the same breast with a previously detected DCIS. Average follow-up interval for the 18 women not developing invasive carcinoma was 16 years. The invasive carcinomas presented clinically from three to ten years (average, 6.1) after the biopsies demonstrating DCIS. Four women with invasive carcinoma developed distant metastases following mastectomy. This study suggests that 28% of women treated with biopsy only for DCIS presenting as an incidental histologic finding will develop invasive carcinoma in a follow-up period of approximately 15 years. PMID- 6275979 TI - Chromate lung cancer with special reference to its cell type and relation to the manufacturing process. AB - The manufacturing process of the chromate industry can be classified into five steps. The main hazardous pollutants of the first step are trivalent chromate compounds, and those of the second and third steps are hexavalent chromate compounds. Analysis of the detailed working history of chromate workers with bronchogenic carcinoma, yielded the following results: the work history of chromatic lung cancer patients in the first step, which produced mainly trivalent chromate dust, was rather short. Patients with small cell carcinoma were engaged mainly in the second step (98.1% of the total working months), where they were heavily exposed to hexavalent chromate dusts. Patients with squamous cell carcinoma were engaged rather evenly in the second and fourth steps, and somewhat longer in the third step. Comparisons of the exposure periods revealed that the working history of small cell carcinoma patients was significantly shorter than that of squamous cell carcinoma patients. There were many more heavy smokers among the squamous cell carcinoma group as compared to the small cell carcinoma patients. Based on these results, it is concluded that the cell type of occupational lung cancer was mainly small cell carcinoma when the exposure to carcinogenic agents was heavy, and that the carcinogenicity of chromate was derived mainly from hexavalent chromate rather than trivalent compounds. PMID- 6275980 TI - Effects of tocopherol (vitamin E) acid succinate on morphological alterations and growth inhibition in melanoma cells in culture. AB - The effects of various forms of tocopherol (vitamin E) on the growth and differentiation of mouse melanoma (B-16) and mouse fibroblast (L-cells) cells in culture were studied. D-alpha-tocopherol acid succinate induced morphological alterations and growth inhibition in melanoma cells. When vitamin E acid succinate was removed 4 days after treatment, the above changes remained irreversible for a period of 24 hr, after which resistant cells and partially affected cells renewed cell division and eventually reached confluency. The relative efficacy of D and DL forms of vitamin E acid succinate remains to be evaluated. However, other forms of vitamin E such as DL-alpha-tocopherol free alcohol, Aquasol DL-alpha-tocopherol acetate, DL-alpha-tocopherol nicotinate, or sodium succinate with an equivalent volume of ethanol, at similar concentrations, were ineffective. Vitamin E acid succinate at similar concentrations did not induce morphological changes in fibroblasts. Melanoma cells were about 2-fold more sensitive to vitamin E acid succinate than were fibroblasts for the criterion of growth inhibition. Vitamin E acid succinate-induced morphological changes and growth inhibition in melanoma cells were expressed in hormone supplemented serum-free medium, but the concentration requirement was about 5 times less than that needed in serum-supplemented medium. Although cyclic adenosine 3': 5'-monophosphate-stimulating agents are known to cause growth inhibition and morphological changes in melanoma cells in culture, vitamin E acid succinate-induced morphological alterations in melanoma cells are no mediated by a rise in cellular cyclic adenosine 3':5'-monophosphate. Ethanol was sufficient to increase the melanin content in melanoma cells. These data show that vitamin E acid succinate may be a potentially useful tumor therapeutic agent. PMID- 6275981 TI - Organization and expression of endogenous virus-like (VL30) DNA sequences in nontransformed and chemically transformed mouse embryo cells in culture. AB - A cloned mouse DNA fragment containing an endogenous "virus-like" DNA (VL30 DNA) sequence was identified by virtue of its ability to hybridize to the virus-like RNA component of mixed-pseudotype AKR-murine leukemia virus virions, its lack of detectable sequence homology with cloned AKR-murine leukemia virus DNA, and its hybridization to a 5.6 kilobase pair (30S) cellular polyadenylic acid [poly(A)] containing RNA species. Restriction enzyme mapping of the cloned mouse fragment revealed the presence of a 5- to 6-kilobase pair VL30 DNA segment flanked by non VL30 segments of approximately 7 and 0.3 kilobase pairs. Southern blot analysis of VL30 DNA sequence organization in the DNA of two nontransformed mouse cell lines (AKR-2B, C3H/10T 1/2) and two chemically transformed derivatives (AKR-MCA, C3H/MCA-58) revealed 15 to 20 bands organized in an apparent strain-specific pattern. Within a given strain, however, no differences were detectable between the nontransformed cells and their chemically transformed counterparts. The expression of VL30 genes in the above cell lines was assayed by hybridization of 32P-labeled poly(A)-containing polysomal RNA to several internal restriction fragments derived from the cloned VL30 DNA sequence. The level of VL30 RNA was enhanced approximately 10-fold in both chemically transformed cell lines as compared to the nontransformed cell lines (under normal growth conditions). In addition, nontransformed AKR-2B cells maintained in the presence of purified epidermal growth factor exhibited similarly enhanced levels of VL30 RNA sequences in polysomal RNA. Since these cells displayed several growth characteristics of transformed cells but, in an epidermal growth factor-dependent and completely reversible fashion, these data suggest that the expression of VL30 genes is not simply incidental to chemically transformed cells but may be related to alterations in growth control. PMID- 6275982 TI - Mouse embryonic transforming growth factors related to those isolated from tumor cells. AB - Growth factors with the two characteristic properties of sarcoma growth factor, the ability to stimulate anchorage-independent growth of normal mouse or rat fibroblasts and the ability to compete with 125I-labeled epidermal growth factor for receptor binding, can be isolated from 12- to 13-day-old normal mouse embryos of various strains. Two size classes of the transforming factor from embryo cells can be isolated with apparent molecular weights of 20,000 and 10,000. The lower molecular-weight factor has been purified several hundred-fold and has the same properties as the peptides with a molecular weight of 10,000 produced by mouse sarcoma virus-transformed 3T3 cells. Whole-mouse embryos contain approximately 10% as much sarcoma growth factor per g of tissue as do the murine sarcoma virus transformed cells; how it is distributed among the embryonic tissues remains to be determined. PMID- 6275983 TI - Correlation between the loss of the transformed phenotype and an increase in superoxide dismutase activity in a revertant subclone of sarcoma virus-infected mammalian cells. AB - We have studied the effects of paraquat (methyl viologen), a herbicide that increases intracellular production of superoxide radical, on the viability of virus-transformed and nontransformed normal rat kidney (NRK) cells in culture. We have shown that a low concentration of paraquat (12.5 microM) is cytotoxic toward virus-transformed cell lines, including Kirsten sarcoma virus- and SV40 transformed NRK cells. The corresponding untransformed NRK cells were resistant to the same and a 4-fold higher concentration of paraquat. There was a good correlation between the susceptibility of transformed and untransformed cells to paraquat cytotoxicity and their ability to increase the superoxide dismutase (SOD) enzymatic activity. We found that paraquat is cytotoxic toward Kirsten sarcoma virus-transformed and SV40-transformed NRK cells which showed low intracellular SOD activity. The relationship between SOD activity and paraquat cytoxicity was strengthened by the finding that the tolerance of NRK cells to the drug was associated with high intracellular SOD activity. This report also describes the isolation of a revertant (revertant RE8G3) cell line derived from Kirsten sarcoma virus-transformed NRK cells after paraquat treatment which contains SOD activity at levels much higher than those found in NRK cells. This revertant is undistinguishable from NRK cells with respect to its lack of transformed cell properties. Not only are these cells normal morphologically but also they do not grow in soft agar, an in vitro property that closely correlates with in vivo tumorigenicity. Several biological and biochemical properties of RE8G3 cells, including growth characteristics, surface receptors for both transferrin and epidermal growth factor (EGF), and the EGF-dependent 32P phosphorylation of specific membrane polypeptides have been studied. The most interesting conclusion that can be drawn from these studies is that there is a correlation between loss of the transformed phenotype and an increase in both EGF receptors and EGF-dependent 32P phosphorylation of a m.w. 170,000 membrane associated protein. PMID- 6275984 TI - Block in the expression of differentiation markers of rat thyroid epithelial cells by transformation with Kirsten murine sarcoma virus. AB - Well-differentiated epithelial cells, derived from primary cultures of normal rat thyroid glands (T-79 cells), as well as a cloned cell line also derived from normal rat thyroid glands (FRT-L cells) were infected with Kirsten murine sarcoma virus carrying outer coat of the helper Kirsten murine leukemia virus. Infected T 79 and FRT-L cells changed morphologically and began to proliferate rapidly, suggesting malignant transformation by the virus. Both cell lines can support the replication of both transformation-competent and transformation-incompetent viruses such as murine or rat leukemia viruses. Infected T-79 and FRT-L cells had a high colony-forming efficiency (68 and 64%, respectively) when grown in agar and formed tumors when transplanted s.c. into syngeneic rats. These tumors morphologically resemble undifferentiated adenocarcinomas, thus showing that Kirsten sarcoma virus carrying the outer coat of the helper Kirsten murine leukemia virus is able to transform differentiated epithelial cells. Transformed T-79 and FRT-L cells, in contrast to uninfected cells, neither secrete thyroglobulin concentrate iodide, two biochemical markers of differentiated thyroid function. Thus, expression of the differentiated phenotype is blocked as a consequence of cell transformation. The system described may be useful in studying epithelial cell carcinogenesis in terms of regulated expression of differentiated functions. PMID- 6275985 TI - Metabolism and binding of benzo(a)pyrene and 2-acetylaminofluorene by short-term organ cultures of human and rat bladder. AB - The ability of organ cultures of normal human and rat bladder to metabolize the polycyclic hydrocarbon, benzo(a)pyrene (BP), and the arylamine, 2 acetylaminofluorene, has been studied. Cultures were maintained for 0 to 6 days in a chemically defined medium before incubation with [3H]BP (0.3 to 0.5 microM) or 2-[14C]acetylaminofluorene (18 to 25 microM) for 24 hr. Ethyl acetate-soluble and water-soluble metabolites were produced from both compounds by both species. The ethyl acetate extracts from [3H]BP-treated human cultures contained 9,10 dihydro-9,10-dihydroxybenzo(a)pyrene, 7,8-dihydro-7,8-dihydroxybenzo(a)pyrene, and 3-hydroxybenzo(a)pyrene. Rat bladder cultures produced similar metabolites but in slightly different proportions. Ethyl acetate-soluble products of 2 [14C]acetylaminofluorene from human cultures contained 7-hydroxy-2 acetylaminofluorene, 9-hydroxy-2-acetylaminofluorene, 2-aminofluorene, and N hydroxy-2-aminofluorene. Rat bladder cultures produced similar metabolites, but 2 aminofluorene was found in relatively higher proportion. Hydrolysis by beta glucuronidase of the water-soluble products produced from both carcinogens gave ethyl acetate-extractable derivatives. These hydrolyzable glucuronide conjugates were relatively more abundant following metabolism of the carcinogens by the rat than by the human cultures. Covalent binding to DNA occurred with [3H]BP in both human (19.7 +/- 13 pmol/mg DNA) and rat cultures (22.8 +/- 8.6 pmol/mg DNA). As with other human tissues, considerable variation (50-fold) was observed between individuals. The results demonstrate that both human and rat bladder epithelium can metabolize known potent carcinogens and, in the case of BP, can effect covalent binding between the products of metabolism and the urothelial cell DNA. In theory, carcinogenesis in the urinary bladder could thus be initiated by carcinogens produced or excreted in the urine without the necessity for their prior metabolism elsewhere in the body. PMID- 6275986 TI - Methotrexate in the chemotherapy of lung cancer. AB - Methotrexate (MTX) is an active agent in both non-small cell and small cell anaplastic lung cancer. A 26% partial response rate was obtained in 27 previously untreated patients with squamous cell bronchogenic tumors treated with 24-hour infusions of MTX at 400 mg/m2. Responses were seen predominantly in extrathoracic sites. In 38 evaluable patients with small cell carcinomas, 24-hour infusions of MTX at 200 mg/m2 in combination with cyclophosphamide and CCNU produced a 43% complete remission (CR) rate for a median survival of 62 weeks. Fifty-six percent of the CR's were seen in patients with extensive disease. PMID- 6275987 TI - Combination chemotherapy for small cell carcinoma of the lung: continuous versus alternating non-cross-resistant combinations. AB - After stratification for extent of small cell lung cancer, 109 patients were randomized to receive cycles of chemotherapy with cyclophosphamide, doxorubicin, and VP-16-213 [CAVP16 (regimen I)] or to receive CAVP16 to maximum response (minimum of three courses) and then chemotherapy with CCNU, methotrexate, vincristine, and procarbazine (COMP) alternating with CAVP16 (regimen II). A group of patients who achieved complete remission were randomized to receive whole-brain irradiation or to have observation only. Of the 44 patients with limited disease, 28 (64%) achieved a complete remission and 11 (26%) achieved a partial remission. Of the 65 patients with extensive disease, 26 (40%) achieved a complete remission and 28 (46%) achieved a partial remission. There were no significant differences between the regimens in response or survival. The projected median survival times are 14 and 10 months for limited and extensive disease, respectively. Nearly 30% of patients with limited disease will be 2 year, disease-free survivors. Twenty-nine patients were randomized to receive cranial irradiation or observation only; none of the 15 irradiated patients developed cerebral metastases, but five of 14 randomized to observation relapsed in the brain (P = 0.02). One patient died with necropsy evidence of only intracranial disease. The principal hematologic toxic effect was leukopenia. There were 31 febrile episodes (21 infectious) during neutropenia and four toxic deaths. Nonhematologic toxicity was mild. Cranial irradiation in patients who achieve complete remission delays or reduces the incidence of CNS metastases. Although alternating chemotherapy is not beneficial, combination chemotherapy with CAVP16 alone is highly effective treatment modality for small cell. PMID- 6275988 TI - Phase II study of procarbazine in small cell carcinoma of the lung. AB - Thirty-nine patients with small cell anaplastic carcinoma of the lung were given oral doses of procarbazine ranging from 100 to 200 mg/m2 on Days 1-14 every 4 weeks. All patients had previously received combination chemotherapy. No responses were observed in the 24 evaluable patients. In reviewing the literature, we were unable to find evidence that the inclusion of procarbazine in combination chemotherapy is beneficial in small cell anaplastic carcinoma. Accordingly, it is concluded that procarbazine should not be considered as a useful drug in the management of this cell type of lung cancer. PMID- 6275989 TI - Phase I-II study of isotopic immunoglobulin therapy for primary liver cancer. AB - A phase I-II study of isotopic immunoglobulin therapy was performed in 18 patients with primary liver cancer; 14 were evaluable for toxicity. The patients received a dose of 37-157 millicuries of 131I-labeled antibody. The dose-limiting factor appears to be hematologic toxicity, especially thrombocytopenia. An objective antitumor effect was seen in six of nine patients who were evaluable for response. Present results suggest that further clinical studies with isotopic immunoglobulin are indicated. PMID- 6275990 TI - Biochemical activation of AZQ [3,6-diaziridinyl-2,5-bis(carboethoxyamino)-1,4 benzoquinone] to its free radical species. AB - The aziridinyl quinone 3,6-diaziridinyl-2,5-bis(carboethoxyamino)-1,4 benzoquinone is an antitumor agent that enhances electron flow from reduced nicotinamide adenine dinucleotide phosphate (NADPH) to molecular oxygen in rat liver microsomes, rat liver nuclei, and purified NADPH-cytochrome c reductase. The process is enzymatic and has an optimum pH of 7.5; NADPH is the preferred cofactor. Electron spin resonance studies show the presence of a free radical semiquinone characterized by five hyperfine lines at g = 2.0046. PMID- 6275991 TI - Relationship of 5'-nucleotidase activity and antileukemic effect in 2' deoxycoformycin therapy. PMID- 6275992 TI - Influence of propranolol isomers and atenolol on myocardial cyclic AMP, high energy phosphates and vulnerability to fibrillation after coronary artery ligation in the isolated rat heart. AB - The isolated rat heart with ligation of the left coronary artery was used to assess the role of the beta 1- adrenergic receptor-cyclic AMP mechanism in the genesis of vulnerability to ventricular fibrillation in early myocardial ischaemia. Coronary artery ligation was followed after 3 min by a reduction in ventricular fibrillation threshold which reached a minimum at 15 min. This was accompanied by reduction of ATP and phosphocreatine while cyclic AMP was significantly increased in ischaemic myocardium. The dl-, l- and d-isomers of propranolol attenuated the decrease in ventricular fibrillation threshold and the increase in ischaemic myocardial cyclic AMP, without altering the tissue depletion of ATP. Specific beta 1-adrenergic receptor antagonism with atenolol did not prevent either the increase of tissue cyclic AMP or the reduction in ventricular fibrillation threshold and high energy phosphates. These findings suggest that the mechanism whereby vulnerability to fibrillation is increased in very early myocardial ischaemia is linked to changes in cyclic AMP content of ischaemic myocardium and appears independent of depletion of myocardial high energy phosphates. PMID- 6275993 TI - Problems of serious and nosocomial infections. Symposium. PMID- 6275994 TI - Doses of antibiotic in patients with renal insufficiency. AB - The pharmacokinetic properties of cefotaxime were studied in 30 subjects with various degrees of renal function after a single 1-gm intramuscular injection. Serum and urinary concentrations of cefotaxime and desacetyl cefotaxime were determined by high-pressure liquid chromatography. The pharmacokinetic parameters were obtained using a one-compartment open model. A significant (P less than 0.0001) linear correlation was demonstrated between the elimination rate constant of cefotaxime and creatinine clearance. The mean serum half-life of cefotaxime was 0.86 hour in normal subjects and was prolonged to 2.35 hours in hemodialysis patients. The mean 24-hour urinary recovery was 51.7% as cefotaxime and 25.6% as desacetyl cefotaxime in normal subjects. In the present study, modification of the dosage is probably unnecessary in patients with creatinine clearance of more than 30 ml/min; however, dose reduction or prolongation of dosage interval, or both, may be appropriate in patients with severe renal dysfunction. PMID- 6275995 TI - Antibiotic treatment of urinary tract infections. AB - Cefotaxime was compared with ceftezole for clinical efficacy and safety in the treatment of chronic complicated urinary tract infections. Cefotaxime was significantly superior to ceftezole in both overall clinical and bacteriological efficacy. There was no significant intergroup difference in adverse reactions or laboratory parameters. Cefotaxime, therefore, proved to be of considerable value in the treatment of chronic complicated urinary tract infections. PMID- 6275996 TI - Experience with cefotaxime in gynecology and obstetrics. AB - Minimal inhibitory concentrations of cefotaxime, determined against clinical isolates from gynecological and obstetrical infections, were less than or equal to 1.56 microgram/ml against about 65% of the aerobes tested. Against anaerobic organisms, the growth of about 65% of the strains was inhibited by less than or equal to 12.5 microgram/ml. The passage of cefotaxime into human uterine tissue was investigated after a single 1-gm intravenous (IV) injection. Peak concentration of 12.24 microgram/gm was attained after about 20 minutes. Passage to the pelvic cavity fluid was studied after a single 2-gm IV dose of cefotaxime. Peak concentration of 22.8 microgram/ml was obtained after about 80 minutes. The placental passage of cefotaxime after a single 1-gm IV injection also was investigated. Compared with the mother's serum concentration, cord serum concentration was about one fourth and amniotic fluid concentration about one tenth. A maximum concentration of 0.63 microgram/ml was seen in the neonate. Clinical response rates were extremely favorable: 98% against uterine infections, 94% against adnexitis, 91% against pelvic infections, and 93% against external genital infections. Bacteriological effectiveness by pathogen was likewise high, showing eradication rates of 95% against gram-positive aerobes, 88% against gram negative aerobes, and 96% against gram-negative anaerobes. PMID- 6275998 TI - Bacteremia in the 1980s with special reference to the role of cefotaxime. PMID- 6275997 TI - Clinical experience in the diagnosis and treatment of infections in the compromised host. AB - A number of disorders of host resistance to infections occur in cancer patients. Defects in granulocytes, complement synthesis and antibody formation, cellular immunity, and mucocutaneous boundaries are discussed. Most infections in these compromised patients are caused by a few gram-negative rods and gram-positive cocci, although other bacteria, opportunistic fungi, intracellular parasites, and DNA viruses also cause severe disease. Cefotaxime was found to be an effective antimicrobial in the treatment of 40 cancer patients with infections of the urinary or lower respiratory tracts, bacteremia, soft-tissue infections, and fever of undetermined origin. Cefotaxime in combination with ticarcillin or an aminoglycoside may improve response in infections due to opportunistic organisms such as Pseudomonas. PMID- 6275999 TI - Clinical comparison of cefotaxime versus the combination of gentamicin plus clindamycin in the treatment of peritonitis and similar polymicrobial soft-tissue surgical sepsis. PMID- 6276000 TI - Clinical evaluation of cefotaxime in the treatment of purulent meningitis in children. PMID- 6276001 TI - Microcrystal arrangement in human deciduous dental enamel studied by electron paramagnetic resonance. PMID- 6276002 TI - Adenosine 3',5'-monophosphate-dependent protein kinases from mammalian artery: isolation and properties of the isoenzymes. PMID- 6276004 TI - Crystalloid inclusions in the Sertoli cell of the koala, Phascolarctos cinereus (Marsupialia). AB - Crystalloid inclusions are a common feature in the basal region of Sertoli cells in the koala, Phascolarctos cinereus. Generally located near the nucleus, they are non membrane-bounded, slender rectangular structures composed to tubules which are orientated at right angles to the long axis of the crystalloid and regularly arranged in rows parallel to this long axis. The tubules in adjacent rows are offset from one another at definite angles and extensively interconnected by filaments. Neither the composition nor function of the crystalloids has been determined, but their association with tonofilaments and the presence of ribosomes in the vicinity suggests that they are most likely proteinaceous. PMID- 6276005 TI - Intramitochondrial crystalloids in rat pinealocytes. PMID- 6276003 TI - Lack of correlation between ultrastructural and pharmacological types of non adrenergic autonomic nerves. AB - In order to test the premise that non-adrenergic, non-cholinergic (NANC) autonomic nerves have a distinctive ultrastructural appearance, clearly different from that of cholinergic nerves, a detailed quantitative ultrastructural analysis has been made of the non-adrenergic innervation of 15 tissues thought from pharmacological evidence to be innervated by NANC nerves (rat and rabbit anococcygeus muscles; rabbit hepatic portal vein; extrinsically denervated toad lung); cholinergic nerves (atria of rat, rabbit, guinea-pig and toad); or both (guinea-pig cervical and thoracic trachealis muscle; rabbit rectococcygeus muscle; urinary bladder of rat, rabbit, guinea-pig and toad) in addition to their adrenergic supply. Following fixation with a modified chromaffin procedure allowing identification of adrenergic nerves, large, randomly selected samples of non-adrenergic nerve profiles from each tissue were analysed with respect to numbers, relative proportions, and size frequency distributions of different vesicle classes within the profiles. The neuromuscular relationships within each tissue were also analysed. On the basis of these analyses, it is clear that there are no consistent ultrastructural differences between cholinergic and NANC autonomic nerves: neither proportions nor sizes of the vesicles provide any clue as to the transmitter used by a particular nerve. The great majority of nerve profiles, whether cholinergic or NANC, contain predominantly small clear "synaptic" vesicles. Large filled "peptidergic" vesicles are no more common in most NANC PMID- 6276006 TI - T antigen binding and the control of SV40 gene expression. PMID- 6276007 TI - Theiler's virus persists in glial cells during demyelinating disease. AB - Theiler's murine encephalomyelitis virus (T-MuEV) is the agent of a persistent, demyelinating infection of the central nervous system of mice, T-MuEV RNA was detected in histological sections of brain and spinal cord of experimentally infected animals by in situ hybridization. Both neurons and glial cells contained viral RNA during the early acute phase of the disease, The amount of viral RNA in neurons, however, was considerably higher than in glial cells. During the late demyelinating phase of the disease, viral RNA was found in low amounts only in glial cells of the white matter of spinal cord. At that stage, no viral RNA was found in neurons. These results demonstrate that T-MuEV persists in glial cells of the white matter. A reconstruction of the pathogenesis of this persistent infection is proposed, based on the different levels of virus replication in neurons and glial cells. PMID- 6276008 TI - Generation of BALB-MuSV and Ha-MuSC by type C virus transduction of homologous transforming genes from different species. AB - The nature of the cell-derived (bas) sequences of BALB-MuSV, a spontaneous mouse sarcoma virus isolate, was determined. Molecularly cloned bas sequences demonstrated no detectable homology with the onc genes of other mouse transforming viruses, but exhibited a high degree of sequence homology with the ras gene of the rat-derived Harvey murine sarcoma virus (Ha-MuSV) genome. The Ha MuSV cell-derived sequence (ras) shared a colinear 750 bp region of homology with bas. Moreover, BALB-MuSV transformation was associated with the expression of high levels of a 21,000 dalton protein, immunologically related to the ras gene products, p21. Thus bas and ras represent retroviral transforming gene homologs that were independently transduced by mouse type C viruses from the genomes of different species. PMID- 6276009 TI - Formation of infectious progeny virus after insertion of herpes simplex thymidine kinase gene into DNA of an avian retrovirus. AB - We have prepared several infectious stocks of an avian retrovirus, spleen necrosis virus, containing the herpes simplex virus type 1 thymidine kinase (tk) gene. The viruses were produced after cotransfection of chicken cells with DNA from recombinants between cloned spleen necrosis virus and tk DNAs and DNA of cloned reticuloendotheliosis virus strain A. removal of sequences in the tk gene for the end of tk mRNA increased a thousand fold the yield of infectious recombinant virus. Infection of chicken or rat tk- cells with the recombinant virus transformed them to a tk+ phenotype. PMID- 6276010 TI - Abelson leukemia virus induces lymphoid and erythroid colonies in infected fetal cell cultures. AB - An examination of Abelson murine leukemia virus (A-MuL V)-hematopoietic cell interaction in cultures of fetal tissues reveals that A-MuLV can stimulate the formation of two different types of colonies. One type of colony is white and composed of A-MuLV-transformed lymphoid cells that can develop into established cell lines. These cells are indistinguishable in morphology from typical adult derived lymphoid transformants. The second type of colony is pink or red and composed of erythroid cells in various stages of differentiation. Although A-MuLV is required to induce the erythroid colonies, and at least some cells in all of these colonies are infected with the virus, no permanently growing cell lines have been established from the cells in these colonies. The frequency of the two types of colonies varies depending upon the tissue and the gestational age of the embryo. Erythroid colonies are found following infection of early and mid gestation tissues while lymphoid colonies are found following infection of mid and late gestation tissues. Mixing experiments indicate that the two types of colonies arise from distinct target cells. Because A-MuLV mutants that are defective for lymphoid cell transformation are also defective for erythroid colony induction, expression of a functional Abelson protein is probably required for colony induction. Thus A-MuLV is capable of stimulating the cells of two distinct hematopoietic lineages. In one case, infection leads to transformation, while in the second, it leads to growth and differentiation. Both types of interaction are mediated, at least in part, by the same A-MuLV gene product, a molecule previously considered to induce transformation in all stably infected cells. PMID- 6276011 TI - Harvey and Kirsten sarcoma viruses promote the growth and differentiation of erythroid precursor cells in vitro. AB - Harvey and Kirsten murine sarcoma viruses have previously been shown to transform fibroblastic cells in culture, and type C virus pseudotypes of these viruses cause erythroleukemia in susceptible mice. We report a cell culture assay for quantitating the growth-promoting effect of Harvey and Kirsten viruses on erythroid cells. Murine hemopoietic cells were infected in vitro with Harvey or Kirsten sarcoma virus, and then cultured in methylcellulose in the presence of relatively low concentrations of erythropoietin. Under these conditions, large colonies of erythroid cells form in the semi-solid culture media 6 to 8 days after infection. The induction of erythroid bursts was not caused by the murine type C helper viruses used to pseudotype either Ha-MuSV or Ki-MuSV, or by media from cells carrying the Ki-MuSV and Ha-MuSV genomes. Induction of the erythroid colonies is under genetic control at the Fv1 susceptibility locus, but not at the Fv2 susceptibility locus. A striking feature of the erythroid colonies induced by the Harvey and Kirsten viruses was that they not only proliferated to large size but also differentiated along the erythroid lineage and synthesized hemoglobin. The results indicate that Ha-MuSV and Ki-MuSV can induce proliferation of erythroid precursor cells apparently without interfering with the differentiation program of the cells. The relation between the growth-promotion effect of these viruses on erythroid precursor cells and their ability to induce erythroleukemia is discussed. PMID- 6276012 TI - The Zea mays mitochondrial gene coding cytochrome oxidase subunit II has an intervening sequence and does not contain TGA codons. AB - Cross hybridization between maize mitochondrial DNA fragments and a specific yeast mitochondrial DNA probe from the oxi 1 gene has been used to identify and isolate the maize mitochondrial gene coding cytochrome oxidase subunit II, mox 1. The DNA sequence reveals two coding regions separated from each other by a single centrally located intervening sequence. Hybridization of mox 1 DNA probes to mitochondrial RNA from plants shows that te gene is transcribed and indicates that several transcripts are spliced. TGA codons, which code Trp in the mitochondria of all species examined to date, do not occur in this gene. However, alignment of the mox 1 gene sequence with the amino acid sequences of subunit II from other organisms strongly suggests that codon CGG (normally Arg) codes for Trp in maize mitochondria, in addition to the standard Trp codon TGG. PMID- 6276013 TI - Highly conserved GC-rich palindromic DNA sequences flank tRNA genes in Neurospora crassa mitochondria. AB - In sequencing a 2200 bp region of the Neurospora crassa mitochondrial DNA encoding the 3' end of the large rRNA gene and a cluster of six tRNA genes, we have found that the tRNA genes are flanked by highly conserved GC-rich palindromic DNA sequences. An 18 bp long core sequence, 5'-CC CTGCAG TA CTGCAG GG 3', containing two closely spaced Pst I sites, is common to all these palindromic sequences. Each of the eight Pst I sites mapped in the 2200 bp region consists of two closely spaced Pst I sites; thus this 2200 bp long segment actually contains 16 Pst I sites. Between 5-10% of the N. crassa DNA may consist of these GC-rich palindromic sequences that include the 18 base long core sequence. The same core sequence is present within both the 5' and 3' side of the intervening sequence of the large rRNA gene, close to, but not at, the intron-exon boundaries. We discuss probable roles for these sequences in N. crassa mitochondrial function, including their role as signals either in the synthesis or processing (or both) of RNA in the mitochondria. PMID- 6276014 TI - Loss of rDNA methylation accompanies the onset of ribosomal gene activity in early development of X. laevis. AB - The rRNA genes of Xenopus blood cells are heavily methylated, but there are two regions in the spacer that frequently contain unmethylated CpG. The undermethylated regions coincide with two regions containing a 60 nucleotide tandemly repeated sequence, and they are present in all somatic tissues that we have tested. Sperm rDNA, by contrast, is fully methylated at these sites in the spacer, and indirect evidence suggests that this may also be the case in oocytes. Loss of methyl groups occurs progressively over the first 20 hr of development, the same period in which embryonic rRNA synthesis initiates and increases in rate. PMID- 6276015 TI - Ty elements are involved in the formation of deletions in DEL1 strains of Saccharomyces cerevisiae. PMID- 6276016 TI - An irregular satellite sequence is found at the termini of the linear extrachromosomal rDNA in Dictyostelium discoideum. AB - IN Dictyostelium discoideum most of the rRNA genes are known to be present as linear, extrachromosomal palindromic molecules. Here we report the identification, molecular cloning and partial DNA sequence analysis of the terminal Eco RI restriction fragment (Eco RI 1) of the rDNA. We identify a length heterogeneity confined to the most distal region of uncloned Eco RI 1, and demonstrate, by DNA sequence analysis of five independently isolated recombinant plasmids containing the Eco RI 1 fragment, that the heterogeneity can be accounted for by variable tracts of an unprecedented irregular satellite sequence with the general formula [CnT]m; n varies from 1 to 8 and accounts for the irregularity, while m varies from 18 to 34 and accounts for the length heterogeneity. The proximal [CnT]m pattern is identical (albeit irregular) in all five clones of Eco RI 1, while the distal [CnT]m sequence appears to vary between clones. Immediately proximal to the CnT satellite are found four nearly perfect tandem repeats of a 29 bp sequence. We discuss the evolution of these simple sequences at the rDNA termini and speculate about possible functions for the CnT satellite. Using the cloned terminal restriction fragment as probe, we also show that virtually all the rDNA in Dictyostelium exists in the linear palindromic form. PMID- 6276017 TI - Identifying P factors in Drosophila by means of chromosome breakage hotspots. AB - A syndrome of germline abnormalities in Drosophila melanogaster called hybrid dysgenesis is thought to be caused by transposable genetic elements known as P factors. Several lines of evidence presented here show that the chromosomal positions of at least some P factors can be identified as points of frequent chromosome breakage (hotspots). Starting with a strain (pi 2) in which four hotspots had been identified on the X chromosome, we found individual hotspots vanished when their part of the chromosome was replaced by the homologous part from a strain known to lack P factors. All hotspots in the non-substituted parts of the chromosome remained functional, indicating that they can act autonomously. We also observed a new breakage site coinciding with the appearance of an unstable mutation at the singed bristle locus (snW). This mutation was dysgenesis induced, and previous genetic evidence suggested that it was caused by the insertion of a P factor at that locus. We also present preliminary evidence for rapid scrambling of the positions of hotspots under certain conditions, and we describe a new procedure for efficiently determining the positions of hotspots on a given chromosome. PMID- 6276018 TI - Analysis of transforming gene products from Moloney murine sarcoma virus. AB - We previously showed that in vitro translation of M-MuSV virion RNA yielded a 62 kd gag gene product and an overlapping set of four proteins with approximate molecular weights of 37,000, 33,000, 24,000 and 18,000. In this paper we show, by use of hybrid arrest translation with cloned recombinant DNAs containing M-MuSV v mosMo sequences, that the 37, 33, 24 and 18 kd proteins are synthesized in their entirety from the v-mosMo gene. Analysis of the primary sequence of these proteins shows that each one is initiated independently from AUG codons within the v-mosMo gene and utilizes the long open reading frame predicted from the v mosMo DNA sequence. Antisera against synthetic peptides corresponding to the C terminus of the predicted v-mosMo gene product precipitate all four in vitro v mosMo proteins. PMID- 6276019 TI - A cascade of adenovirus early functions is required for expression of adeno associated virus. AB - One measurable biological activity of early adenovirus genes is their ability to promote growth of the defective adeno-associated virus, AAV. We have identified an ordered sequence of communications among the early genes of adenovirus type 2 (Ad2) that results in expression of the helper activity. We purified DNA fragments and mRNAs corresponding to early Ad2 regions E1, E2A, E3 and E4 and injected them via glass capillaries into AAV-infected cells. DNAs were placed in the nucleus, mRNAs in the cytoplasm. AAV DNA and proteins synthesized in response to the injected Ad2 nucleic acids were extracted from as few as 100 cells and identified by gel electrophoresis. Our results reveal a cascade of early Ad2 gene regulation, E1 leads to E2A leads to E4, with E4 providing the helper effect for AAV. PMID- 6276020 TI - Genetic analysis of crown gall: fine structure map of the T-DNA by site-directed mutagenesis. PMID- 6276021 TI - The remarkable instability of replication loops provides a general method for the isolation of origins of DNA replication. AB - Closed superhelical replicative intermediates of SV40 DNA were stable during incubation at elevated temperatures for prolonged times. However, when the replicative intermediates were nicked or made linear by digestion either outside or inside of the replication loop, the nascent DNA strands were spontaneously extruded at a measurable rate even at 37 degrees C. The extruded DNA was double stranded. The rate-limiting step in the extrusion was not the rate of branch migration. The instability of replication loops suggested a method for the isolation of replication origins. The method has been tested by a reconstruction experiment in which a small amount of linear SV40 replicative intermediates was mixed with a vast excess of cellular DNA. The mixture was heated overnight at 55 degrees C, and the spontaneously extruded nascent strands were isolated and inserted into pBR322. Plasmids containing SV40 DNA of less than genomic size, symmetrically distributed about the SV40 replication origin, were isolated. This result suggests that the method is generally applicable to the isolation of replication origins. PMID- 6276022 TI - Somatic expression of herpes thymidine kinase in mice following injection of a fusion gene into eggs. AB - A plasmid, designated pMK, containing the structural gene for thymidine kinase from herpes simplex virus (HSV) fused to the promoter/regulatory region of the mouse metallothionein-I gene, was injected into the pronucleus of fertilized one cell mouse eggs; the eggs were subsequently reimplanted into the oviducts of pseudopregnant mice. The first experiment produced 19 offspring, one of which expressed high levels of HSV thymidine kinase activity in the liver and kidney. pMK DNA sequences were detected in equal amounts in several tissues of the expressing mouse as well as in three mice that did not express HSV thymidine kinase activity. In all cases, several copies of the pMK plasmid were tandemly duplicated and integrated into mouse DNA. It appears as though multiple copies of the intact plasmid were fused by homologous recombination either before or after integration at a single site in the mouse genome. The overall efficiency of obtaining somatic expression of thymidine kinase in experiments performed to date is about 10% (4/41), and twice this number have integrated pMK DNA. This procedure not only provides a means of introducing new genes into mice, but it will also be a valuable system for studying tissue-specific regulation of gene expression. PMID- 6276023 TI - In vitro mutation analysis of the mating-type locus in yeast. AB - The mating-type locus (MAT) of Saccharomyces cerevisiae is a complex locus that codes for the regulators of cell type. Two unique messages are transcribed from each MAT allele. Using the in vitro mutagenesis technique whereby synthetic oligonucleotides containing restriction sites (linkers) were inserted into plasmids, we have constructed a series of mutations in cloned DNA containing either the MATa or MAT alpha locus. The new restriction site associated with each "linker" mutation has allowed the mutation to be mapped and sequenced. We have complemented genetically defined mutations (mata1, mat alpha 1 and mat alpha 2) with plasmids containing these in vitro mutations by yeast transformation, thereby mapping the genes onto the DNA sequence. MATa1 has been localized to the MATa unique region (Ya) from which the a1 message is transcribed. We find no function for the other MATa message by using our complementation assay. MAT alpha 1 maps to the MAT alpha unique (Y alpha) and adjacent (Z) region from which the alpha 1 message is transcribed. MAT alpha 2 maps to the other major message found in the common (X) region of the MAT alpha loci. Although most linker mutations that have a mutant phenotype appear to disrupt the translated portion of each gene, two mutations may disrupt transcription. PMID- 6276024 TI - A 200 base pair region at the 5' end of the chicken adult beta-globin gene is accessible to nuclease digestion. AB - A DNA segment close to the 5' end of the chicken adult beta-globin gene contains a hypersensitive site for nuclease action. Using a variety of endonucleases, we show that this hypersensitive site is in fact an accessible region, which extends from approximately 60 to approximately 260 base pairs 5' from the start of mRNA transcription. The presence of this hypersensitive region is correlated with gene activity; it is observed in nuclei isolated from the definitive erythrocytes of 14-day chicken embryos, but is not observed in nuclei isolated from the primitive erythrocytes of 5-day embryos (which do not express this gene) or in brain nuclei or purified DNA. The close spacing of accessible digestion sites on this DNA segment is not consistent with the presence of a normal nucleosome. Furthermore, a 115 base pair DNA fragment contained within this nuclease-sensitive region can be excised by Msp I digestion and released from nuclei, in at least 50% yield. Approximately one third of the released fragments behave as protein-free DNA. PMID- 6276025 TI - Inhibition of plasminogen activator release from transformed chicken fibroblasts by a protease inhibitor. AB - Chicken embryo fibroblasts (CEF) transformed by Rous sarcoma virus (RSV) exhibit increases in both a cell-associated and a secreted form of plasminogen activator (PA). The mechanism whereby the membrane-bound, cell-associated form of PA is processed to an extracellular, soluble form has been examined in cultures of chicken fibroblasts transformed by a temperature-sensitive mutant of RSV. We report that chymostatin, a protease inhibitor of limited specificity, inhibits the release of PA from tsRSVCEF while causing accumulation of cell-associated PA. Chymostatin's effect on PA release is specific, reversible and appears to be due to its anti-proteolytic capacity. Chymostatin does not inhibit cellular protein synthesis or interfere in the assay used to measure PA. A chymostatin-sensitive protease activity has been found in a membrane fraction isolated from tsRSVCEF. PMID- 6276026 TI - 5',3' Cyclic adenosine monophosphate dependent protein phosphorylation in differentiating male mouse germ cells. PMID- 6276027 TI - Microtubules and the regulation of DNA synthesis in fibroblastic cells. (A minireview). PMID- 6276028 TI - Localization of the regulatory subunit of type II cyclic AMP-dependent protein kinase on the cytoplasmic microtubule network of cultured cells. PMID- 6276029 TI - Coupling of chromatin structure to cell geometry during the cell cycle: transformed versus reverse-transformed CHO. PMID- 6276030 TI - Histiocytic differentiation of mouse malignant fibrous histiocytoma-like tumor cells by culturing with sodium butyrate. PMID- 6276031 TI - [Scleromalacia perforans (author's transl)]. PMID- 6276032 TI - Comparison of the photodynamic action of Rose Bengal and tetrachlorosalicylanilide on isolated porcine erythrocyte membranes. AB - The light-mediated effects of Rose Bengal and 3,3',4',5-tetrachlorosalicylanilide (T4CS) on porcine erythrocyte membranes have been investigated. Irradiation in the presence of Rose Bengal and oxygen causes extensive destruction of the unsaturated fatty acids of the erythrocyte membrane. This results in a decrease in the membrane flexibility as measured by a nitroxide spin probe. Irradiation in the presence of T4CS and oxygen had no measurable effect on the unsaturated fatty acids or on the membrane flexibility. Irradiation of erythrocyte membranes both in the presence of Rose Bengal and oxygen and of T4CS gave rise to polymerisation of the membrane proteins. This was evident by polyacrylamide gel electrophoresis and by freeze-fracture electron microscopy. Aggregation of membrane proteins could be observed with low levels of Rose Bengal and short irradiation times at which no loss of unsaturated fatty acids could be detected. Irradiation of the n butanol-extracted apoprotein of porcine erythrocyte membranes both in the presence of Rose Bengal and of T4CS resulted in polymerisation of the protein as measured by gel electrophoresis and electron microscopy. The results obtained from the two photodynamic compounds are compared in terms of their different mechanisms of action on the membrane. The implications of the results in determining the molecular events leading to photohaemolysis are discussed. PMID- 6276033 TI - Tissue-specificity of N-nitrosodibutylamine metabolism in Sprague-Dawley rats. AB - The distribution and metabolism of N-[14C]nitrosodibutylamine were studied in Sprague-Dawley rats. The results indicated that in addition to the liver, metabolism of the substance occurred in the nasal mucosa, the lung and the oesophagus. Metyrapone and diethyldithiocarbamate reduced the production of 14CO2 from N-[14C]nitrosodibutylamine by all these tissues. There was no indication of metabolic capacity in the urinary bladder or the kidney. The results fit with the assumption that tumours of the urinary tract are induced by metabolites reaching these tissues via the urine. Besides the liver, the oesophagus and the lung are target tissues for the carcinogenicity of N-nitrosodibutylamine in Sprague-Dawley rats and in these tissues the local formation of reactive metabolites may play a role in the pathogenesis of N-nitrosodibutylamine-induced lesions. PMID- 6276034 TI - [A microwave unit for killing experimental animals]. PMID- 6276035 TI - [Study on virus yields in Igarashi clone of Singh's Aedes albopictus cells persistently infected with Japanese encephalitis virus (2-8 strain)]. PMID- 6276036 TI - [Clinical and electroencephalographic analysis of 41 cases of infantile spasm with report on 4 cases]. PMID- 6276037 TI - [Relationship between PGs (E1 and F2 alpha) and cAMP in plasma and urine of chronic nephritis and Shen-Hsu according to the differential diagnosis of traditional Chinese medicine (author's transl)]. PMID- 6276038 TI - [Studies on the morphogenesis of herpes simplex virus. I. Electron microscopic observation on its cytoplasmic morphogenesis]. PMID- 6276039 TI - [2 cases of granular cell myoblastoma of the esophagus]. PMID- 6276040 TI - [Association of carcinosarcoma of the endometrium and ductal carcinoma of the breast. Presentation of a case]. PMID- 6276041 TI - ELISA--a simple test for detecting and differentiating antibodies to closely related orthopoxviruses. AB - In this paper, the results are reported of experiments to investigate the optimum conditions for determining, and differentiating between, the antibodies against closely related poxviruses using the ELISA technique. This assay was found to be considerably more sensitive than the virus neutralization, haemagglutination inhibition, passive haemagglutination, and indirect fluorescent antibody tests for detecting antibodies, but has not yet been compared with radioimmunoassay. The use of ELISA made it possible to differentiate between antibodies to vaccinia, whitepox, and monkeypox viruses. PMID- 6276042 TI - Shortening of the action potential and reduction of pacemaker activity by lidocaine, quinidine, and procainamide in sheep cardiac purkinje fibers. An effect on Na or K currents? PMID- 6276043 TI - Physician practice in the management of patients with uncomplicated myocardial infarction: changes in the past decade. AB - To determine changes in the patterns of care between 1970 and 1980 for patients with uncomplicated acute myocardial infarction, questionnaires were sent to almost 6000 physicians in 1979 and responses were compared with those of a similar survey taken in 1970. Almost all physicians in 1979 reported the availability and use of an intensive care/coronary care unit facility with continuous electrocardiographic monitoring. Progressive-care facilities are also becoming more widely available. The median length of hospitalization has decreased markedly. Early ambulation and an earlier return to work are more common. There is a high level of informal patient and patient-family counseling about myocardial infarction and its management, both during and after hospitalization, and wider use of educational materials. Most physicians continue to recommend progressive physical activity after hospitalization. The routine prescription of anticoagulant therapy during hospitalization has declined, while prescription of prophylactic antiarrhythmic agents has increased. Nitrate drugs and tranquilizers are routinely prescribed by a large percentage of physicians for their patients with uncomplicated myocardial infarction. Use of standard exercise tests has increased among all physician specialties. The treadmill test is most often used, and testing is typically done 6 weeks after infarction. A significant increase in the availability of and familiarity with exercise testing is characteristic of all medical specialties. Symptoms of new chest pain and palpitations are now considered important enough to warrant the recommendation to report immediately to an emergency room. Other current findings include the routine use of coronary angiography by a large percentage of physicians to evaluate the need for surgical intervention, and the routine posthospitalization prescription by only a small percentage of physicians of aspirin and of nitrate drugs for patients with uncomplicated myocardial infarction. PMID- 6276044 TI - Optimal resources for the care of patients with acute myocardial infarction and chronic coronary heart disease. Coronary Heart Disease Study Group. PMID- 6276045 TI - Enzymic measurement of urinary pyrophosphate with a centrifugal analyzer. AB - We describe a simple, rapid, and fully automated technique for measuring urinary pyrophosphates with a centrifugal analyzer (the ENI GEMSAEC). This technique depends on the enzymic magnesium-dependent reaction with UDPG pyrophosphorylase (UTP: alpha-D-glucose-1-phosphate uridylyl transferase, EC 2.7.7.9) and spectrophotometry of the NADPH formed in a combined system of phosphorylation and reduction. Many samples of urine can be analyzed quickly without pretreatment, with high sensitivity (1.3 mA/mumol of substrate) and good reproducibility. The mean within-run coefficient of variation for a 50 mumol/L pyrophosphate solution was 1.4%. We determined the optimum enzyme and magnesium concentrations necessary for use in a 4-min reaction. Because there is no inhibitory effect of chloride and phosphate ions, pyrophosphate can be measured directly in urine, without prior extraction. With this technique, the mean value (and SD) for urinary pyrophosphate excretion by 30 healthy subjects was 39.3 (SD 17.2) mumol/24 h. PMID- 6276046 TI - cAMP radioimmunoassay without interference from calcium or EDTA. AB - Calcium and EDTA interfere with radioimmunoassay of cAMP or acetylated cAMP. Both interferences with the assay of acetylated cAMP can be obviated by including in the buffer 15 mmol of CaCl2 per liter, which at the same time increases the affinity of the antibody by 89%. As a result, antibody can be decreased by 65%. This method requires no prepurification (extraction or chromatography), and no correction for analytical recovery is necessary. This assay can detect 12 fmol of cAMP per tube, so only microliter quantities of plasma are needed. PMID- 6276047 TI - Differential pulse polarography of the catalytic waves in serum from patients with urological cancers. AB - Catalytic waves obtained from the perchloric acid-soluble fraction of serum in the presence of hexaminecobalt trichloride appear to conform to a Langmuir adsorption isotherm. Wave heights for sera from patients with untreated urological cancers significantly exceed those from normal subjects. In general, treatment with diethylstilbestrol decreased the wave height. Effects of some other chemotherapeutic agents on wave heights are also noted. PMID- 6276048 TI - Clinical significance of plasma galactose and erythrocyte galactose-1-phosphate measurements in transferase-deficient galactosemia and in individuals with below normal transferase activity. AB - We correlated the clinical symptoms of transferase-deficient galactosemia with the plasma galactose and erythrocyte galactose-1-phosphate concentrations in six galactosemic patients during dietary treatment, in a child before treatment, and in 12 individuals with below-normal erythrocyte hexose-1-phosphate uridylyltransferase activity. All the treated patients were asymptomatic. Normal galactose and either normal or above-normal galactose-1-phosphate concentrations were found. Three of these patients were clinically normal as newborns while ingesting galactose-containing foods and may resemble the asymptomatic Negro galactosemic. The clinical symptoms of galactosemia were observed in the untreated patient, who showed markedly above-normal concentrations of galactose and galactose-1-phosphate, protein and reducing substances in the urine, above normal bilirubin and alkaline phosphatase in the plasma, with normal values for glucose, aspartate aminotransferase, alanine aminotransferase, and gamma glutamyltransferase. Clinical improvement in this patient paralleled the decline in erythrocyte galactose-1-phosphate. The individuals with below-normal hexose-1 phosphate uridylyltransferase activity (range 7--17 U/g of hemoglobin) had normal galactose and galactose-1-phosphate concentrations and were asymptomatic. PMID- 6276049 TI - Ovarian carcinoma; second-look laparotomy postchemotherapy. Preliminary report. PMID- 6276050 TI - Measurements of angiotensin converting enzyme in captopril-treated patients. PMID- 6276051 TI - Bromocriptine suppresses ACTH secretion from human pituitary tumour cells in culture by a dopaminergic mechanism. AB - Bromocriptine (0.13-13 microM) significantly inhibited ACTH secretion in a dose dependent manner when added to cell cultures of a human corticotrophic adenoma for 24 h. Haloperidol (13 microM), but not serotonin (13 microM), blocked this inhibition but had no significant effect when added alone. In addition, dopamine (10 microM) reduced ACTH secretion during a 4-h incubation, whereas serotonin (0.01-10 microM) was ineffective. An ectopic ACTH secreting lung carcinoid was non-responsive to doses of bromocriptine up to 13 microM. These results demonstrate a direct suppressive action of bromocriptine on a human pituitary corticotrophic adenoma through dopaminergic rather than serotoninergic mechanisms. PMID- 6276052 TI - Pituitary - adrenocortical function in patients during treatment with the angiotensin-converting enzyme inhibitor captopril. AB - To study effects on pituitary-adrenocortical activity of a sustained block of angiotensin II formation, six 'drug-resistant' patients with essential hypertension were studied before and during treatment with an inhibitor of the angiotensin-converting enzyme (Captopril, SQ 14,225). The drug was given in increasing doses (100-400 mg/day) for 2 weeks whilst patients received a moderately restricted sodium intake (60-80 mmol/day). Immunoreactive ACTH, cortisol, aldosterone, plasma renin activity (PRA) and the activity of the angiotensin-converting enzyme (ACE) were measured in blood samples drawn at 0800 0900 h. Urinary excretion of cortisol and aldosterone were measured in 24-h urine collections. Further information on pituitary-adreno-cortical function was obtained by measuring serial plasma corticosteroid levels after submaximal stimulation with a synthetic ACTH preparation. ACTH and cortisol did not change an observation which does not support the hypothesis that glucocorticoid activity is influenced by a decrease in plasma angiotensin II concentrations. PMID- 6276053 TI - Angiotensin II stimulates the release of ACTH from dispersed rat anterior pituitary cells. AB - In a rat anterior pituitary dispersed cell system, angiotensin II was found to stimulate the release of ACTH at concentrations ranging from 100 pmol/l to 1 mumol/l, with a maximal response being given by 10 nmol/l. The angiotensin II antagonist, saralasin, was able to block completely at a concentration of 1 mumol/l the stimulation of ACTH induced by 10 nmol/l angiotensin II, but had no effect on the basal release. The log dose-response curve for ACTH release by angiotensin II was shifted to the right in a parallel fashion by saralasin 10 nmol/l, suggesting competitive antagonism. The stimulation of ACTH release by a rat stalk-median eminence extract or by arginine vasopressin was unaffected by saralasin. The data are interpreted as suggesting that rat corticotrophs contain angiotensin II receptors, and that these may be involved in ACTH release in response to hypovolaemic stress. PMID- 6276054 TI - Comparison between dose-responses of prolactin, thyroid stimulating hormone and growth hormone to two different histamine H-2 receptor antagonists in normal men. AB - The effects of the histamine H2-receptor antagonists, ranitidine and cimetidine on prolactin (PRL), thyroid stimulating hormone (TSH), and growth hormone (GH) were studied in six normal males. Intravenous bolus injections of saline and of 50, 100, and 200 mg ranitidine and 300 mg cimetidine were tested. Ranitidine (100 and 200 mg) and cimetidine (300 mg) caused a significant increase in PRL secretion, whereas saline and ranitidine (50 mg) did not. TSH and GH secretion were unaffected by all doses. A dose-response relationship between ranitidine and PRL was established, and a dose of 65 mg ranitidine was found to be the minimal effective PRL-releasing dose. Plasma ranitidine concentration was measured by specific RIA. The results indicate an effect of H2-receptor antagonists on physiological functions in the control of PRL secretion. Histamine, therefore, may play a role in the regulation of PRL secretion. PMID- 6276055 TI - [Characterization of insulin-sensitive phosphodiesterase in fat cells (author's transl)]. PMID- 6276056 TI - Genetic analysis of induction of anti-polyadenylic acid antibodies and xenotropic type-C viruses. AB - Normal mice can be induced by lipopolysaccharide to produce anti-polyadenylic acid (poly A) antibodies and xenotropic (X-tropic) type-C viruses. To determine whether these traits are genetically linked, high anti-poly A antibody, high virus-producing NZB mice were crossed with low anti-poly A antibody-producing, virus-negative SWR mice. All mice in the F1 generation could be induced by LPS to produce high levels of both anti-poly A antibodies and X-tropic virus. When F1 hybrids were back-crossed to the SWR parent, all offspring were high anti-poly A antibody producers, but one-third of the mice remained virus-negative. These results demonstrate that these two LPS responses of a B lymphocyte are not genetically-linked and the anti-poly A antibody response is multigenic. PMID- 6276057 TI - Increased conversion of arachidonic acid to vasodilator prostanoids in spontaneously hypertensive rats. AB - 1. Vasodepressor responses to intravenous injection of prostacyclin, arachidonic acid, and nitroprusside were examined in anaesthetized, spontaneously hypertensive rats (SHR) of the Okamoto strain, and in their normotensive Wistar Kyoto (WKY) controls. 2. Depressor responses to prostacyclin and nitroprusside did not differ significantly between the two strains. 3. The vasodepressor effects of arachidonic acid were greater and much more prolonged in SHR than in WKY. In rats treated with indomethacin (2 mg/kg) arachidonic acid induced only transient depressor responses which did not differ significantly between these strains. 4. It is concluded that SHR do not differ from WKY in their sensitivity to prostacyclin but they have enhanced ability to transform exogenous arachidonic acid into vasodilator prostanoids. PMID- 6276058 TI - Effect of converting enzyme inhibition with captopril on in vitro angiotensin production in sheep. AB - 1. The effect of captopril on in vitro production of angiotensin I (ANG I), [Val5]-angiotensin II ([Val5]-ANG II) and [Val4]-angiotensin III ([Val5]-ANG-(2 8)) in central venous blood taken from sodium-deficient sheep was studied. 2. Captopril enhances in vitro production of ANG I but blocks the in vitro production of [Val5]-ANG II and [Val5]-ANG-(2-8). 3. The production of ANG I in blood is faster than that of [Val5]-ANG II and [Val5]-ANG-(2-8). 4. The half-life of [Val5]-ANG II and [Val5]-ANG-(2-8) in vitro in blood in the presence of captopril was 10 and 14 min, respectively. 5. This in vitro study suggests that the production of [Val5]-ANG II and [Val5]-ANG-(2-8) in blood forms a small part of the total body production of each peptide. PMID- 6276059 TI - Immunoreactive beta-endorphin levels in plasma and pituitary tissue from genetically hypertensive and normotensive rats. AB - 1. Levels of immunoreactive beta-endorphin were measured in neurointermediate lobes, anterior lobes and plasma from the Japanese and New Zealand strains of genetically hypertensive rats and their normotensive controls. 2. No significant differences were observed in beta-endorphin between the hypertensive and normotensive rats of the New Zealand strain. 3. The hypertensive rats of the Japanese strain showed significantly higher levels of beta-endorphin in neurointermediate lobe and lower levels in plasma than their normotensive controls. 4. These results suggest that the differences in beta-endorphin levels in the Japanese strain reflect a genetic difference not necessarily related to elevated blood pressure. PMID- 6276060 TI - Hypertension through adrenaline activation of prejunctional beta-adrenoceptors. AB - 1. Adrenaline can enhance the stimulation-induced release of transmitter noradrenaline in sympathetically innervated tissues by activating prejunctional beta-adrenoceptors. 2. Adrenaline incorporated into sympathetic transmitter stores by neuronal uptake can be subsequently released as a co-transmitter and can then activate prejunctional beta-adrenoceptors, thus completing a facilitatory feedback loop. 3. Rats chronically treated with adrenaline develop elevated blood pressures compared to control rats. beta-Adrenoceptor blockade prevents the rise in blood pressure. 4. Activation by adrenaline of facilitatory prejunctional beta-adrenoceptors of sympathetic nerves innervating cardiovascular effector tissues may explain adrenaline-induced rises in blood pressure. PMID- 6276061 TI - Production of angiotensin converting enzyme by cultured bovine endothelial cells. AB - 1. Endothelial cells from bovine aortae were grown to confluence in tissue culture. The cells were subcultured and angiotensin converting enzyme (ACE) production was measured while the cells were maintained in serum-free medium for 4 days. 2. During this period, there was a ten-fold increase in ACE concentration in the medium whereas the cellular content of the enzyme remained constant at approximately 23% of the total enzyme produced. Cycloheximide (1 mumol/l) blocked production of ACE. 3. The enzyme liberated into the medium closely resembled ACE in the following respects: it cleaved the dipeptide histidyl-leucine from a synthetic tripeptide substrate, was markedly Cl- activated and inhibited by EDTA (1 mmol/l), SQ 14225 (1 mumol/l) or SQ 20881 (1 mumol/l). 4. Cells maintained in culture for up to ten passages have maintained morphology typical of endothelial cells and continue to produce ACE. PMID- 6276062 TI - Dual effects of clonidine on rat prejunctional alpha-adrenoceptors. AB - 1. The effects of clonidine (1.0 mumol/l) and phentolamine (3.0 mumol/l) on the stimulation-induced efflux of radioactivity were assessed in rat isolated atria and tail arteries in which transmitter stores had been labelled with (3H) noradrenaline. 2. In atria, clonidine did not affect transmitter release when stimulation was for 10 s periods at frequencies of 1, 2 or 5 Hz, whereas at 10 Hz there was a significant 1.2-fold enhancement of release. Phentolamine enhanced release with stimulation at 5 Hz and 10 Hz by 1.5- and 2.5-fold respectively. 3. In tail arteries, clonidine reduced release when stimulation was for 30 s periods at a frequency of 1 Hz, but did not affect release at 2 Hz or 5 Hz. Phentolamine in each case produced an approximately 1.5-fold enhancement of release. 4. In the pithed rat, spinal stimulation of cardioaccelerator fibres produced a tachycardia, the magnitude of which depended on the frequency and duration of stimulation. Clonidine (10 micrograms/kg, i.v.) caused either a reduction or an enhancement of the response depending on the frequency and duration of stimulation; in the presence of phentolamine (2 mg/kg, i.v.), clonidine had no effect. 5. The results are consistent with partial agonistic actions of clonidine at prejunctional alpha-adrenoceptors in the rat cardiovascular system; such effects may be of relevance to the mechanism of the clonidine withdrawal syndrome. PMID- 6276064 TI - Regulation of uterine smooth muscle bradykinin receptors by bradykinin levels and angiotensin converting enzyme inhibitor in the rat. AB - 1. Bradykinin infusion (0.1 microgram/min i.v.) decreased the number of uterine bradykinin receptors by 20% at Day 2. Bradykinin receptors returned to control levels at Day 7. 2. Captopril infusion (1.7 micrograms/min i.v.) induced prolonged decreases in the number of uterine bradykinin receptors of 15% at Day 2 and of 13% at Day 7, respectively. 3. The number of uterine bradykinin receptors was increased in two-kidney, one clip hypertensive rats by 19%. 4. These results suggest that endogenous bradykinin participates in the regulation of uterine bradykinin receptors. 5. Decreased uterine bradykinin receptors induced by captopril might reflect increased endogenous bradykinin. PMID- 6276063 TI - Role of the kallikrein-kinin system in the renal effects of angiotensin converting enzyme inhibition in anaesthetized dogs. AB - 1. Administration of captopril (1.5 mg/kg i.v.) to anaesthetized dogs was associated with an increase in renal blood flow of 56 ml min-1 (s.e.m. = 13, n = 9) despite a significant fall of 17 mmHg (s.e.m. = 5, n = 9) in mean arterial pressure. 2. Treatment of dogs with the angiotensin receptor antagonist, Sar1 Ile8-angiotensin II (2.5 micrograms/kg per min i.v.), or the cyclo-oxygenase inhibitor indomethacin (10 mg/kg i.v.) did not prevent the renal vasodilation and hypotension following angiotensin-converting enzyme inhibition. This suggests that these effects are neither solely due to inhibition of the renin-angiotensin system nor mediated by prostaglandins. 3. Increased urinary kinin excretion, possibly reflecting increased renal concentrations of kinins, accompanied the renal vasodilation after both captopril and renal artery occlusion. 4. The kallikrein-kinin system may play a role in the regulation of the renal vasculature in anaesthetized dogs. PMID- 6276065 TI - Haemodynamic response to ACTH administration in essential hypertension. AB - 1. The haemodynamic and volume response to ACTH administration was investigated in six patients with mild, untreated essential hypertension and two patients with Addison's disease on maintenance steroids. Blood pressure, heart rate and weight were recorded daily. Plasma volume (125I-HSA) and cardiac output (thermo dilution) were measured during the control period and on the 5th day of ACTH treatment. 2. In the hypertensive subjects, mean arterial pressure rose from 94.3 +/- 2.2 to 105.7 +/- 2.8 mmHg on the 5th day of ACTH administration (P less than 0.02). Plasma volume rose from 29.8 +/- 2.2 to 34 +/- 2.2 ml/kg. Cardiac index increased from 2.85 +/- 0.21 to 3.32 +/- 0.14 l/min per m2 (P less than 0.05). Cardiac output rose from 5.81 +/- 0.69 to 6.72 +/- 0.59 l/min. Calculated total peripheral resistance, heart rate and body weight were unchanged. No such changes were seen in patients with Addison's disease. 3. The haemodynamic characteristics of ACTH in patients with mild untreated essential hypertension are similar to those in the experimental model of ACTH induced hypertension in sheep. PMID- 6276066 TI - [A case of neuronal ceroid-lipofuscinosis (Jansky-Bielshowsky type): morphological, biochemical and electrophysiological studies (author's transl)]. AB - We reported a case of late infantile neuronal ceroid-lipofuscinosis. The patient was an 8-year-old boy presenting with marked psychomotor deterioration, progressive visual failure due to retinal degeneration and optic atrophy, startle reaction to auditory stimuli, frequent myoclonus and generalized convulsions. The routine laboratory examinations were all normal. EEG was markedly abnormal because of poorly organized background activity and frequent paroxysmal spike-and wave complexes. CT scan showed evidence of severe atrophy of the cerebrum, cerebellum and brainstem. Electron microscopic examination of the biopsied rectum revealed fingerprint profiles in the neurons and pericytes beneath the muscularis mucosa. Cultured skin fibroblasts also contained electron dense inclusions, some of which showed fingerprint profiles. Urinary glycopeptides were normal. Lyscsomal enzyme activities in leukocytes and cultured fibroblasts were normal. Neurophysiological studies revealed giant cortical potentials evoked by the auditory as well as somatosensory stimulation. Simultaneous recording of the somatosensory evoked EEG and EMG potentials disclosed that the myoclonus in this patient was stimulus-sensitive and compatible with the cortical reflex myoclonus. With regard to hypothetical pathogenesis of this disease, we studied lipoperoxide in the blood before and after anti-oxidant therapy. We also measured vitamin A and carotene, since these substrates are related to retinoic acid. Although vitamin A and carotene were normal, lipoperoxide was slightly elevated. However, it was not influenced by the treatment with anti-oxidant. Significance of elevated lipoperoxide to the pathogenesis of this diseases has not solved. PMID- 6276067 TI - [Fine structures of Marinesco body and nuclear body in the substantia nigra (author's transl)]. PMID- 6276069 TI - The effect of carotid endarterectomy on mental functioning. AB - In order to ascertain the effect of carotid endarterectomy on mental functioning, carotid endarterectomy patients suffering from transient ischemic attacks (TIA) were compared preoperatively and postoperatively with cholecystectomy patients. Preoperatively, a tendency toward poorer verbal memory and greater well-being is found in carotid endarterectomy patients. Postoperatively, the overall mental functioning of the carotid endarterectomy patients is not significantly improved, though verbal fluency does show improvement. The well-being of both groups is significantly greater following operation. The complaints of the carotid endarterectomy patients during the (last) TIA had already diminished before operation and after operation remained practically on the same level. Preoperatively and postoperatively, no clear connection exists between mental functioning and age of patients in this experiment, not between mental functioning and the side of the operation. Finally, in the case of 5 patients, no long-term (4.5 months) effect of carotid endarterectomy could be indicated. PMID- 6276068 TI - Electrophoresis of cerebrospinal fluid proteins. PMID- 6276070 TI - Cardiac neuropathy in chronic alcoholics. AB - Measurement of beat-to-beat variations was employed in a group of 14 chronic alcoholics and compared with a group of normal individuals. A significant increase in the number of runs was found in the alcoholics reflecting impaired neurogenic control of the heart. It is concluded that cardiac neuropathy should be considered as a contributing factor to the development of alcoholic cardiomyopathy. PMID- 6276071 TI - Unexpected trends in the analysis of a questionnaire and interview procedure to detect transient cerebral ischemic attack in a female population. AB - By means of a case-control study, social and biomedical factors were considered in relation to transient cerebral ischemic attack occurrence as determined by questionnaire and interview among a population of females ( greater than 50 years of age) residing in the city of utrecht and surroundings. Contrary to earlier expectations, a greater TIA risk was found associated with: lower systolic and diastolic blood pressure, a low-salt diet for high blood pressure, lower body weight, smaller body surface, and use of medicines for cardiac insufficiency/arrhythmias and chronic anxiety. The importance of hemodynamic processes (i.e. hypotensive episodes of cerebral circulation) for TIA within the general population is emphasized. PMID- 6276072 TI - Astrocytomas involving the whole spinal cord. Two case reports. AB - We present two patients with an astrocytoma, involving the whole spinal cord. Operation was performed in both cases: one patient died because of complications after the operation, the condition of the other patient improved greatly. In the literature we could find three other cases of astrocytoma, extending into more than ten segments of the spinal cord. The clinical picture and the differential diagnosis with ependymomas of the same length are discussed. PMID- 6276073 TI - External ophthalmoplegia, juvenile parkinsonism and axonal polyneuropathy in two siblings. AB - In one of two siblings a clinical disorder was described, consisting of a slowly progressive juvenile parkinsonism with extensor plantar responses, external ophthalmoplegia with severe ptosis and a motor and sensory polyneuropathy. The younger sibling had only juvenile parkinsonism, Unilateral ptosis and a motor and sensory polyneuropathy. Their father was neurologically normal except for a unilateral ptosis. There did not seem to be consanguinity in this family. PMID- 6276074 TI - Clinical significance of basal ganglia calcifications detected by ct (a retrospective study of 33 cases). AB - In a retrospective study 2015 CT scans were studied for the presence of basal ganglia calcifications. These were discovered in 33 cases. No abnormalities in serum calcium or phosphate levels or any other specific clinical symptoms related to these calcifications were found. It is concluded that basal ganglia calcifications casually detected by CT generally have no clinical significance. PMID- 6276075 TI - Refractory anaemia in the glucagonoma syndrome. PMID- 6276076 TI - Atypical lymphadenopathies of the head and neck. AB - Atypical lymphadenopathies fail to achieve the morphologic criteria of a malignant neoplasm, but exceed the usual concepts of follicular, lymphoid, or sinus histiocytic hyperplasias. Rich cellular proliferations usually composed of prominent histiocytes, or immunoblasts, or both with or without a vascular scaffolding obscure the nodal architecture. Toxoplasmosis, infectious mononucleosis, zoster, and vaccination-induced lymphadenopathies are caused by infectious agents, dermatopathic lymphadenitis is associated with cutaneous disease, anticonvulsant pseudolymphoma occurs in individuals hypersensitive to anticonvulsants (usually phenytoin), and Chediak-Higashi syndrome is an inherited abnormality of lysosomal microtubule function; the causes of sinus histiocytosis with massive lymphadenopathy, giant lymph node hyperplasia, angioimmunoblastic lymphadenopathy, mucocutaneous lymph node syndrome, and this histiocytoses remain unknown. The clinical course of these abnormalities varies from self-limited acute diseases (viral lymphadenopathies, toxoplasmosis, dermatopathic lymphadenitis, and usually anticonvulsive lymphadenopathy) to protracted, but benign abnormalities (sinus histiocytes with massive lymphadenopathy, giant lymph node hyperplasia, and multifocal eosinophilic granuloma). The diagnosis of angioimmunoblastic lymphadenopathy, Chediak-Higashi syndrome, and mucocutaneous lymph node syndrome necessitates a guarded prognosis, for death or the advent of a malignant lymphoma may interrupt their clinical course. Acute disseminated histiocytosis, even though the proliferated cell lacks the cytologic criteria of malignancy, should be regarded and treated as a malignant neoplasm. PMID- 6276077 TI - Effect of preservation at low temperature (4 degrees C) on polymorphonuclear neutrophil integrity as determined by biochemical analysis. PMID- 6276078 TI - Effect of cryogenic preservation (-80 degrees C) on polymorphonuclear neutrophil integrity as determined by biochemical analysis. PMID- 6276079 TI - Glucagonoma tumors and syndrome. PMID- 6276080 TI - Covalent modification of phosphofructokinase by phosphorylation- dephosphorylation. PMID- 6276081 TI - Regulation of liver 3-hydroxy-3-methylglutaryl-CoA reductase. PMID- 6276082 TI - Phosphorylation reactions that influence the activity of elF-2. PMID- 6276083 TI - Mono (ADP-ribosyl)transferases and their effects on cellular metabolism. PMID- 6276084 TI - Glycogen synthase and glycogen synthase kinases. PMID- 6276086 TI - Gene cloning in organisms other than E. coli. PMID- 6276085 TI - Cloning vectors derived from plasmids and phage of Bacillus. PMID- 6276087 TI - Vectors for cloning in yeast. PMID- 6276088 TI - Cloning with 2-micrometer DNA vectors and the expression of foreign genes in Saccharomyces cerevisiae. PMID- 6276089 TI - Selectable markers for the transfer of genes into mammalian cells. PMID- 6276090 TI - Gene transfer into mammalian cells: use of viral vectors to investigate regulatory signals for the expression of eukaryotic genes. PMID- 6276091 TI - Use of plasmids from Staphylococcus aureus for cloning of DNA in Bacillus subtilis. PMID- 6276092 TI - Cauliflower mosaic virus on its way to becoming a useful plant vector. PMID- 6276093 TI - The Ti plasmids of Agrobacterium. PMID- 6276094 TI - Vectors for gene cloning in Pseudomonas and their applications. PMID- 6276095 TI - Host: vector systems for gene cloning in Pseudomonas. PMID- 6276096 TI - Gene cloning in Streptomyces. PMID- 6276097 TI - Gene cloning in Neurospora crassa. PMID- 6276098 TI - The structure and replication of rhinoviruses. PMID- 6276099 TI - Poxvirus DNA. PMID- 6276100 TI - Kaposi's sarcoma in young homosexual men. AB - An outbreak of KS has been observed in young homosexual men. These patients are different from those with classical KS for the following reasons: 1. geographic distribution (clustering in New York and California); 2. age (younger, mean--39 years); 3. higher incidence; 4. sexual preference (homosexual); 5. distribution of skin lesions (face, upper extremities, trunk); 6. lymph node involvement; 7. visceral lesions; 8. associated opportunistic infections (Pneumocystis carinii, toxoplasmosis); 9. history of sexually-transmitted diseases (hepatitis, syphilis, gonorrhea); and 10. aggressive course of the disease. Awareness of these features of the new KS will enable the practitioner to better recognize this important, emerging disease. PMID- 6276101 TI - The structure of human globin messenger RNA: functional, genetic, and evolutionary implications. PMID- 6276102 TI - The clinical relevance of lymphocyte surface markers in leukemia and lymphoma. PMID- 6276103 TI - Hereditary and acquired defects in the pyrimidine nucleotidase of human erythrocytes. PMID- 6276104 TI - Clinical implications of ectopic hormone production in small cell carcinoma of the lung. PMID- 6276105 TI - Isolated pleural effusion in small cell lung carcinoma: favorable prognosis. A review of the Southwest Oncology Group experience. AB - The currently accepted staging system for small cell lung cancer considers patients who present with a pleural effusion as having extensive disease, but no series of such patients has been reported. Between 1974 and 1980, 56 patients with ipsilateral pleural effusion as the only evidence of metastatic spread beyond the primary tumor and regional nodes were place on three consecutive Southwest Oncology Group studies of chemotherapy and radiation therapy for small cell lung cancer. Effusions were cytology-positive in 24: response rates and survival were not different whether cytology was positive or negative. The overall response rate was 77 percent, with 36 percent who achieved complete response, a result comparable to that for patients with limited disease. The survival of effusion "only" patients did not differ significantly among the studies. Median survival of 54 weeks and survival curve for the effusion "only" group as a whole was identical to that of all patients classed as having limited disease by the usual criteria. Long-term disease-free survival was observed just as commonly: 2/17 patients from the first study have disease-free survival at greater than five years, and overall disease-free survival at greater than two years is 22 percent. Performance status does not explain the favorable survival in this subgroup, since only 55 percent of effusion patients were fully ambulatory (comparable to extensive disease patients as a group) and even fully ambulatory patients with extensive disease rarely had disease-free survival greater than two years. Patients with ipsilateral pleural effusion as their only evidence of metastasis should be staged as having limited disease. PMID- 6276106 TI - Metabolic functions of the lung. Of what clinical relevance? PMID- 6276107 TI - Pathogenesis of asthma. Neurophysiology and pharmacology of bronchospasm. AB - Asthma results from variable and often sudden changes in airway smooth muscle tone. Allergy is not an essential component of the asthmatic response; however, mediator release plays an important role in the human asthmatic response. Histamine may act directly to cause bronchoconstriction by stimulating the H1 receptor on airway smooth muscle or indirectly by stimulation of afferent vagal fibers in airways. Histamine may also act locally on airways to augment cholinergic and, possibly, alpha-adrenergic constrictor effects, or to antagonize beta-adrenergic relaxation of airway smooth muscle. Cholinergic neural output promotes bronchoconstriction in non-atopic asthma, but parasympathetic reflexes are not a major component of human bronchial responses to inhaled allergen. The physiologic significance of the sympathetic nervous system in relaxing airway smooth muscle is incompletely defined. Recent studies suggest that direct sympathetic innervation of airways is relatively unimportant and that purinergic fibers may be the predominant inhibitory neurons in human airways. Investigations focusing on intracellular calcium metabolism in airway smooth muscle have implicated the adenyl cyclase-cyclic adenosine monophosphate system in the regulation of bronchomotor tone. Cyclic nucleotides may modulate but do not mediate respiratory muscle contraction, and their precise role in regulating bronchomotor tone remains uncertain. PMID- 6276108 TI - Hematoporphyrin derivative and laser photoradiation in the treatment of lung cancer. AB - Photoradiation therapy (PRT) was performed in 13 lung cancer cases and in one case of severely atypical squamous metaplasia following administration of hematoporphyrin derivative (HpD). The HpD is activated by visible red light (630 nm, 90 to 400 mW) from an argon dye laser. The cytocidal effects were due to the activation of the HpD, since 400 mW of power has in itself no effect on normal epithelium, even with long-term exposure. HpD is retained longer by malignant tissue than by normal tissue. Therefore, the lesions were irradiated with the red laser beam, delivered by a quartz fiber inserted through the instrumentation channel of the fiberoptic bronchoscope, 48 hours or more after intravenous injection of 2.5 to 4.0 mg/kg of HpD. A total of 14 cases received PRT. In one, two small, smooth-surfaced, squamous cell carcinoma tumors in the right B2b of a 74-year-old man who had refused surgery disappeared three days after HpD photoradiation, and the patient remained disease-free 16 months after the treatment. In 12 cases of centrally located lung cancer local effects were obtained in all. However, there was no significant improvement in survival, attributable to the fact that all were advanced-stage cases. One patient with severely atypical squamous metaplasia requested treatment, and complete disappearance of metaplastic atypic was obtained. PMID- 6276109 TI - An R plasmid of Serratia marcescens transferable to Pseudomonas aeruginosa. AB - Hospital isolates of Serratia marcescens able to transfer resistance to up to 11 antibiotics were found to contain conjugative R plasmids. One set of strains harbors only a single R plasmid with a mass of 89 megadaltons (Mdal). This plasmid codes for resistance to nine antibiotics including ampicillin, carbenicillin, cephalothin, streptomycin, kanamycin, gentamicin, tobramycin, sisomycin, and sulfonamides. The 2nd set of strains harbors 2 R plasmids, 1 with a mass of 89 Mdal, the other 57 Mdal. Analysis of progeny from genetic crosses indicates that the larger R plasmid codes for resistance to the same antibiotics as does the 89-Mdal plasmid described above. The 57-Mdal species codes for resistance to ampicillin, carbenicillin, cephalothin, kanamycin, neomycin, and tetracycline. The 89- and 57-Mdal R plasmids appear unrelated by a number of genetic and physical criteria. The 89-Mdal plasmid, but not the 57-Mdal species, is transferable by conjugation to Pseudomonas aeruginosa, and renders this species stably resistant to carbenicillin, streptomycin, kanamycin, gentamicin, tobramycin, and sisomycin. PMID- 6276110 TI - Cefotaxime (HR-756) in urinary tract infections. AB - A total of 30 hospitalized patients with upper and lower urinary tract infections were treated with cefotaxime (HR-756) by intramuscular or intravenous route. All patients had a monomicrobial infection, and the major part of bacterial isolates were gram-negative bacilli (83%) while gram-positive cocci accounted only for 17%. In 83% of the cefotaxime-treated patients both the bacteria and symptoms were eliminated at the 2nd week of follow-up, while failure or relapse occurred in 17% of the patients. No reinfections were recorded. Cefotaxime was a safe and well-tolerated drug and can be considered as an effective antibiotic in the therapy of urinary tract infections caused by "difficult' gram-negative bacilli. PMID- 6276111 TI - [Tumours of the carotid body: reflections based upon 4 cases including 2 bilateral and occurring in the same family (author's transl)]. PMID- 6276113 TI - [The clinical and pathological features of androblastoma (with report of 7 cases) (author's transl)]. PMID- 6276112 TI - [Diagnosis of granulosa and theca cell tumor of the ovary (with clinical and pathological analysis) (author's transl)]. PMID- 6276114 TI - Functional capacity of suprarenal cortex in patients on long-term dialysis. AB - The authors have repeatedly examined the functional capacity of suprarenal cortex in patients on long-term dialysis by means of ACTH stimulation test. Out of 148 examinations, normal reaction was found in 111 cases, partially insufficient in 16 cases, and fully insufficient in 21 cases. Primary suprarenal insufficiency is considered for the reason of insufficient production of cortisol which develops due to enforced changes of regulatory mechanism in glucocorticoids production as a reaction on repeated long-term losses of cortisol through semipermeable membrane. According to the authors, the mentioned facts are of great significance since they may result in changes which influence the survival of patients on long term dialysis in a negative way. PMID- 6276115 TI - The effect of hypotensive drugs on the intraocular pressure after waterloading in rabbits. AB - The waterloading test in rabbits, using 100 ml of waterloads per kg body weight, without general anaesthesia and with applanation tonometry to measure intraocular pressure (IOP), deserves a place in glaucoma research. Using this model we found adrenaline not to have an effect on the IOP with or without waterloading. Isoprenaline and to a lesser physostigmine reduced the pressure in the normal eye markedly, but these drugs did not have any effect on the pressure after waterloading. delta 9-THC (Tetrahydrocannabinol) and pilocarpine increased the IOP in the normal rabbit eye. After waterloading delta 9-THC decreased the pressure, but its value was still significantly higher than the control pressure. Pilocarpine did not reduce the pressure after waterloading. Acetazolamide and timolol did not reduce the pressure in the normal eye, but after waterloading a significant reduction was observed. PMID- 6276116 TI - Mucinous ovarian tumors with giant cell mural nodules. A report of two cases. AB - Two cases are reported in young women of a variant of ovarian mucinous tumor with giant cell mural nodules. The first tumor was a predominantly proliferating mucinous tumor with a giant cell mural nodule adjacent to foci of haemorrhage and reactive vascular proliferation. No invasive carcinoma could be identified. In the second tumor, which was examined by electron microscopy, a sharp demarcation was observed between typical invasive mucinous carcinoma and areas which, on low power examination, were indistinguishable from malignant giant cell tumor of soft parts. Closer examination revealed, in these latter areas, an intimate mixture of anaplastic, predominantly mononuclear tumor cells and multinucleate osteoclast like giant cells. The histogenesis of these lesions is suggested as being reactive to the tumor cells or their products and is discussed with reference to similar tumors reported from the ovary and other sites in the body. PMID- 6276117 TI - Granular cell myoblastoma of the uterine cervix. PMID- 6276119 TI - [Hormone receptor analysis and cytostatic agent sensitivity testing. Significance in additional therapy planning of breast carcinoma]. PMID- 6276118 TI - Evidence for an endorphin dysfunction in methadone addicts: lack of ACTH response to naloxone. AB - Chronic exogenous opiate administration might be responsible for the acute and protracted abstinence syndrome by producing a prolonged decrease in the availability of endogenous opioids (endorphins). However, the hypothesis that potent exogenous opiates may have anti-endorphin effects has been difficult to test. We have been investigating this hypothesis with neuroendocrine test paradigms which have provided preliminary evidence of anti-endorphin effects for chronic methadone. Naloxone-induced ACTH response data from chronic methadone addicts offers preliminary support for the hypothesis that chronic exogenous opiate administration has anti-endorphin effects. The subjects were 7 male methadone addicts who had been addicted to greater than or equal to 40 mg of methadone and 7 male healthy opiate-naive volunteers. Naloxone failed to produce a significant increase in ACTH in methadone addicts while opiate-naive normal volunteers demonstrated a significant naloxone-induced release of ACTH. Five of the seven methadone addicts ahd no demonstrable ACTH response to naloxone. These impaired naloxone response data reported here for recently detoxified addicts suggest that chronic methadone administration comprises the functional integrity of the endorphin system. Prolonged abstinence, post-detoxification depression and other affective symptoms which contribute to relapse may result from a prolonged endorphin derangement. PMID- 6276120 TI - [Diagnosis of the site of insulinomas: percutaneous transhepatic portal vein catheterisations with selective blood sampling for hormone determination (author's transl)]. AB - Percutaneous transhepatic portal vein catheterisation with blood sampling from the various areas of drainage, especially the pancreatic veins, was undertaken in seven patients with insulinoma to diagnose its site. In six patients measurement of serum-insulin levels revealed an abrupt rise in the vascular area later found to drain the area of the insulinoma. Insulin measurement in one patient with insulinoma in the head of the pancreas falsely indicated an islet-cell tumour in the region of the tail of the pancreas. C-peptide concentration in serum followed the concentration of insulin, but did not show such a marked rise. The method of percutaneous transhepatic portal vein catheterisation with selective blood sampling for the measurement of hormonal concentration was superior to ultrasound, computer tomography or coeliacography for determining the site of the tumour. PMID- 6276121 TI - [Therapy of soft tissue neoplasms]. PMID- 6276123 TI - Ontogeny of 3H-diazepam binding sites in different rat brain area. Effect of GABA. AB - The ontogenesis of the 3H-diazepam binding sites and their modulation by gamma aminobutyric acid have been studied in different brain areas of rat at various ages, using frozen and two Triton X-100 treated crude synaptic membrane preparations. Benzodiazepine recognition sites are present at birth in all the brain regions investigated and reach adult levels about 3 weeks later. The changes of 3H-diazepam binding with age are due to an increase in the total number of binding sites. In vitro addition of GABA produces, in newborn and adult cerebral cortex, an increase in affinity, but not in number of binding sites for 3H-diazepam. The effect is age dependent. PMID- 6276124 TI - [Membrane research today]. PMID- 6276122 TI - Ketoconazole: a review of its therapeutic efficacy in superficial and systemic fungal infections. PMID- 6276125 TI - [Peptide hormone receptors]. PMID- 6276126 TI - [Lipoprotein receptors]. PMID- 6276127 TI - [Membrane transport of sodium and potassium]. PMID- 6276128 TI - [Morphology of hypertensive cardio- and angiopathies]. PMID- 6276129 TI - Catecholestrogens and release of anterior pituitary gland hormones. I. Luteinizing hormone. AB - We investigated the effects of peripheral administration of 17 beta-estradiol (E2), estrone (E1), and the catecholestrogens, 2-hydroxyestradiol (2-OHE2) and 2 hydroxyestrone (2-OHE1), on anterior pituitary gland LH release in the prepuberal rat. Steroids in oil were injected sc into 25-day-old female and 35- to 40-day old male rats. The injection of E2, E1, or 2-OHE2 caused a surge in serum LH levels in female rats 48 h later, during the after hours. Only E1 induced a LH surge 24 h after injection. The positive effects of 2-OHE2 in the females were only observed if a massive dose was administered, the steroid was injected on 2 consecutive days, or E2 or progesterone was given to 2-OHE2-primed rats. The 2 OHE1 was totally ineffective in causing a serum LH surge under a variety of experimental protocols. In male rats, the injection of any one of the four steroids decreased serum LH levels. Even the injection of E2 or 2-OHE2 for 2 days or the injection of E2 in 2-OHE2-primed rats failed to elevate the serum LH concentration in male rats. The results suggest that 2-OHE2 and E1 could play a role in the preovulatory release of LH in the female; 2-OHE2 and 2-OHE1 could play a role in the negative feedback control of LH release in the male. PMID- 6276130 TI - Catecholestrogens and release of anterior pituitary gland hormones. II. Prolactin. AB - We investigated the effects of the peripheral administration of 17 beta-estradiol (E2), estrone (E1), and the catecholestrogens, 2-hydroxyestradiol (2-OHE2) and 2 hydroxyestrone, (2-OHE1), on anterior pituitary gland PRL release in the prepuberal rat. Steroids in oil were injected sc into 25-day-old female and 35- to 40-day-old male rats. The injection of E2, E1, or 2-OHE2, but not of 2-OHE, caused a surge in serum PRL levels in female rats 48 h later, during the afternoon hours. Only E1 induced a PRL surge 24 h after injection. In male rats, the injection of E1 or 2-OHE2, but not of 2-OHE1, elevated serum PRL levels on a chronic basis. The results suggest that 2-OHE1 plays no discernible role in PRL release in either sex, but that 2-OHE2 might play a role in the tonic release of PRL in the male and in the preovulatory release of PRL in the female. PMID- 6276131 TI - Guanine nucleotide and magnesium ion regulation of the interaction of gonadotropic and beta-adrenergic receptors with their hormones: a comparative study using a single membrane system. PMID- 6276132 TI - An analysis of the sources and quantity of 3,5,3'-triiodothyronine specifically bound to nuclear receptors in rat cerebral cortex and cerebellum. PMID- 6276133 TI - Evidence for involvement of alpha-adrenergic receptors in norepinephrine-induced prostaglandin E2 and luteinizing hormone-releasing hormone release from the median eminence. AB - Studies were conducted to determine if the stimulatory effect that norepinephrine (NE) exerts on the release of prostaglandin E2 (PGE2) and LHRH from the median eminence is mediated by alpha- or beta-adrenergic receptors. Incubation of median eminence fragments from adult male rats with different concentrations of phentolamine, an alpha-receptor antagonist, resulted in a dose-related inhibition of the release of both PGE2 and LHRH induced by NE, with an IC50 of 3.5 X 10(-7) and 0.9 X 10(-7) M for PGE2 and LHRH, respectively. Complete suppression of the NE effects was observed with a phentolamine concentration of 5 X 10(-6) M. This dose also reduced basal PGE2 and LHRH release. In contrast to phentolamine, the beta-receptor antagonist propranolol, tested at a concentration of 5 X 10(-6) M, was completely ineffective in altering the stimulatory effect of NE on either PGE2 or LHRH release. Moreover (and as previously observed for LHRH), blockade of dopaminergic receptors with Pimozide (10(-6) M) failed to inhibit the release of PGE2 induced by NE, thus indicating that the stimulatory effect of NE is not mediated by a dopaminergic mechanism. It is concluded that NE stimulates the release of PGE2 and LHRH from nerve terminals of the median eminence by first interacting with an alpha-adrenergic receptor. PMID- 6276134 TI - Pituitary-adrenal function in rats chronically exposed to cold. AB - Exposure to cold for 2 weeks was used to assess the effects of a sustained stimulus on pituitary-adrenal function in male rats. The diurnal peak in plasma and adrenal corticosterone was advanced by 4 h during the first 24 h of exposure to cold but returned to its usual time (2000 h) by the next day. Plasma ACTH and corticosterone levels were generally greater at all times during the 24-h cycle in animals exposed to cold for up to 2 weeks, with the greatest increase occurring consistently at the time of peak. When rats exposed to cold for 1 week were returned to a normal 24 C environment, plasma corticosterone tended to increase. Plasma ACTH and plasma and adrenal corticosterone responses to a superimposed acute provocative stimulus (ip saline injection) were faster, greater, and more sustained in rats exposed to cold for 3 or 7 days. Similarly, the compensatory adrenal hypertrophy response to unilateral adrenalectomy was greater in cold-exposed rats. Such animals were also more resistant to pituitary adrenal suppression by prednisolone. In contrast, there was no change in the sensitivity of the adrenal to exogenous ACTH. The results suggest that chronic exposure to cold causes a sustained activation of central mechanisms that regulate pituitary ACTH secretion as well as extra-pituitary mechanisms that regulate adrenal size; it reduces the effectiveness of negative feedback mechanisms, but does not alter those involved in the regulation of adrenal rhythmicity or adrenal sensitivity to ACTH. PMID- 6276135 TI - Alterations of brown adipose tissue in genetically obese (ob/ob) mice. II. Studies of beta-adrenergic receptors and fatty acid degradation. AB - beta-Receptor number, norepinephrine-stimulated adenylate cyclase activity and lipolysis, octanoate-induced NAD(P) redox changes, and heat production were studied in brown adipose tissue (BAT) of lean and obese (ob/ob) mice. beta Receptor number was increased 1.54-fold in purified brown adipocyte plasma membrane of ob/ob mice compared to that in lean controls. This increase was reversed by cold adaptation (5 C). Basal and norepinephrine-stimulated adenylate cyclase values were not different in the two groups. Norepinephrine stimulated lipolysis at 10 nM in BAT of lean mice, but only at 10 microM in BAT ob/ob mice. Octanoate produced an increase in the NAD(P) redox state in BAT of lean mice, but it did not modify the NAD(P) redox state in BAT of ob/ob mice. Concomitantly, octanoate increased heat production 3-fold in BAT of lean mice, but did not promote any significant increase in heat production in BAT of ob/ob mice. These two parameters were restored toward values observed in lean mice when the ob/ob mice were adapted to a cold environment. The data indicate that BAT of ob/ob mice exhibits three alterations; one at the level of the beta-receptor, one at the level of the lipolytic response to norepinephrine, and one at the level of fatty acid activation and/or beta-oxidation. PMID- 6276136 TI - Membrane potential changes of mouse adrenal zona fasciculata cells in response to adrenocorticotropin and adenosine 3',5'-monophosphate. AB - ACTH superfused onto mouse adrenal zona fasciculata tissue caused a transient, dose-dependent membrane depolarization. The log of the dose of ACTH was linearly related to the magnitude of depolarization. The onset of depolarization was rapid and dose dependent. Resting membrane potential changes observed after ACTH were blocked by CoCl2 but not tetrodotoxin or 4-aminopyridine, indicating that these depolarizations were dependent primarily on transmembrane Ca++ flux. CoCl2 also significantly blocked ACTH-stimulated adrenal steroid production; 4-aminopyridine had a much smaller and greatly delayed effect, whereas tetrodotoxin had no detectable effect on steroidogenesis. cAMP administration to adrenal zona fasciculata cells elicited transient, dose-dependent membrane depolarizations, which closely resembled those observed after ACTH treatment. In contrast to ACTH, CoCl2 did not block the cAMP-induced depolarization. These and other studies indicate that ACTH initiates a complex series of events by which steroidogenesis is stimulated. One mechanism may involve a change in membrane permeability to Ca++ independently of cAMP generation; a second mechanism may involve the activation of adenylate cyclase which subsequently influences the membrane conductance of the fasciculata cell membrane. PMID- 6276137 TI - Evidence for independent secretion of beta-endorphin immunoreactivity from rat pars distalis in vivo. AB - The distinctive chromatographic patterns of beta-endorphin-like immunoreactivity (beta END-LI) released from rat pars distalis (PD) or pars intermedia in vitro as well as the selective inhibitory effects of dexamethasone on PD were used to determine which lobe secretes beta END-LI into plasma after clonidine administration in vivo. Gel chromatography (Sephadex G-50) indicates that cultured PD cells released two major immunoreactive species which coelute with beta-lipotropin (beta LPH) or beta END standards. Conversely, virtually all of the beta END-LI secreted by neurointermediate lobe [pars intermedia plus pars nervosa (NIL)] cells in vitro resembled beta END in size, while none was detected which cochromatographed with beta LPH. Incubation of PD cells with clonidine (10( 6) M) evoked a 2-fold increase in beta END-LI release, while the drug had no effect on beta END-LI released from cultured NIL cells. Dexamethasone (10(-7) M) inhibited the clinidine-induced release of beta END-LI from PD cells, whereas it did not influence the stimulated release of beta END-LI from NIL by isoproterenol (10(-6) M). In vivo administration of clonidine (0.5 mg/kg, ip, for 15 min) increased total plasma beta END-LI from 0.75 +/- 0.16 to 1.35 +/- 0.18 ng/ml; 85% of this rise corresponded to beta LPH as determined by gel chromatography. Prior administration of dexamethasone (60 micrograms/kg, ip, for 4 h) completely prevented the clonidine-induced release of beta END-LI in vivo. These results demonstrate that clonidine probably acts selectively on the PD in vivo to release pituitary beta END-LI, and further, that PD can release beta END-LI independently of the pars intermedia. PMID- 6276138 TI - Magnesium promotes both parathyroid hormone secretion and adenosine 3',5' monophosphate production in rat parathyroid tissues and reverses the inhibitory effects of calcium on adenylate cyclase. AB - Reduced extracellular Ca2+ is known to promote PTH secretion, while severe Mg2+ depletion has the opposite effect. We have correlated the effects of Mg2+ and Ca2+ on parathyroid hormone (PTH) secretion and cAMP accumulation by rat parathyroid tissues in vitro with the effects of these two metals on adenylate cyclase activity in broken membrane preparations. PTH secretion was maximal at 0.5 mM Ca2+, falling to low levels as the Ca2+ concentration was increased to 2.5 mM. Deletion of Mg2+ from the medium resulted in a marked decrease in PTH secretion at any given Ca2+ concentration. At a constant Ca2+ concentration of 1 mM, both PTH secretion and cAMP production rose to maximal rates as the Mg2+ concentration was increased from 0 to 2 mM. The adenylate cyclase of rat parathyroid membranes was stimulated by both GTP and guanyl-5'-yl imidodiphosphate [Gpp(NH)p]. EDTA-treated membranes could not be stimulated by Gpp(NH)p. Repletion with Mg2+ was more effective than repletion with Ca2+ in restoring responsiveness to the guanine nucleotide. When membranes were maximally preactivated by Gpp(NH)p and then assayed in the presence of variable concentrations of metal ions, enzyme activity was directly inhibited by Ca2+ and stimulated by Mg2+. Adenylate cyclase sensitivity to Ca2+ inhibition was dependent upon the Mg2+ concentration; in the presence of 0.6 mM Mg2+ a 50% inhibition was produced by 0.05 mM Ca2+, while in the presence of 8 mM Mg2+ a 10 fold higher Ca2+ concentration was required for a similar inhibitory effect. The results suggest that Ca2+ may decrease PTH secretion at least in part by a direct inhibition of adenylate cyclase. Mg2+ may promote PTH secretion either by enhancing the activation of adenylate cyclase by endogenous guanine nucleotides or by competing with Ca2+ for binding to a distinct regulatory site on the enzyme. PMID- 6276139 TI - Modifications of the steroidogenic pathway during spontaneous and adrenocorticotropin-induced maturation of ovine fetal adrenal. PMID- 6276140 TI - Study of the direct action of luteinizing hormone-releasing hormone agonists at the testicular level in intact rats treated with an antiluteinizing hormone serum. AB - Is it well known that LHRH agonists can inhibit testicular functions by gonadal desensitization secondary to endogenous LH release and that a direct action at the gonadal level has also been demonstrated. Since an excess of anti-LH serum can be used, as an alternative to hypophysectomy, to neutralize the influence of endogenous LH release, the relative importance of the testis and pituitary gland in the inhibitory effect of a LHRH agonist, [D-Ser-(TBU)6,des-Gly-NH210]LHRH ethylamide (Buserelin), was studied on gonadal gonadotropin receptors in intact adult male rats treated with equine anti-LH or normal horse serum (NHS). A single administration of increasing doses (1-100 ng) of Buserelin leads to a progressive inhibition of testicular LH and PRL receptor levels by 70% and 40%, respectively, in animals injected with NHS. Treatment with the anti-LH serum completely prevents this inhibitory effect of a single dose of the LHRH agonist. After two successive injections of Buserelin in NHS-treated animals, testicular LH receptors are reduced by 70% and 80% with the 100- and 500-ng doses, respectively, while testicular PRL receptors are inhibited by 40-60%. In animals treated with the anti-LH serum, the inhibition of testicular LH receptors is reduced by only 18% (100 ng Buserelin) and 55% (500 ng Buserelin), while the inhibitory effect on PRL receptors is abolished. The present data show that endogenous LH release induced by a single injection of a LHRH agonist plays an essential role in the loss of testicular LH receptors measured 2 days later. Moreover, upon repeated injection of the LHRH agonist, neutralization of endogenous LH release by an anti-LH serum markedly reduces the inhibitory effect of the LHRH agonist on LH receptors, while it completely prevents the effect on PRL receptors, suggesting that the inhibitory effect of the LHRH agonist in the male rat is predominantly due to endogenous LH release rather than to a direct action of the peptide at the testicular level. PMID- 6276141 TI - Effects of growth hormone, prolactin, and placental lactogen on insulin content and release, and deoxyribonucleic acid synthesis in cultured pancreatic islets. AB - The direct effects of human GH (hGH), ovine pituitary PRL (oPRL), and human chorionic somatomammotropin [placental lactogen (hPL)] on the endocrine pancreas were studied in isolated pancreatic islets maintained in tissue culture. Islets of Langerhans were isolated by collagenase treatment of pancreatic tissue obtained from adult NMRI mice and adult or newborn Wistar rats. The islets were maintained for up to 3 weeks in petri dishes containing tissue culture medium RPMI 1640 supplemented with newborn calf serum or normal human serum. The release of insulin during culture and the islet content of insulin, glucagon, and DNA after culture were determined. The DNA synthesis in the newborn rat islets was evaluated by the incorporation of [methyl-3H]thymidine into islet cell DNA. In mouse islets, 1 micrograms/ml hGH, oPRL, or hPL markedly stimulated insulin release during a 2-week culture period and caused a significant increase in the insulin content in the islets after culture. While hGH did not affect the DNA content in adult mouse islets, an increase was observed in adult rat islets after 2-3 weeks of culture. In islets isolated from 3- to 5-day-old rats cultured for 2 weeks with hGH, there was a 30-40% higher DNA content than that found without hGH. Correspondingly, a significant stimulation of the incorporation of [methyl 3H]thymidine could be demonstrated 24 h after the addition of hGH, oPRL, or hPL. hCG and porcine ACTH had no effect. In conclusion, these results indicate that GH and related hormones have a direct stimulatory effect on both the insulin production and DNA synthesis in isolated islets of Langerhans. Whether the effect is directly on the beta-cell or mediated via locally produced growth factors remains to be determined. PMID- 6276142 TI - Growth hormone receptors in isolated rat adipocytes. AB - Specific GH binding sites in isolated rat adipocytes have been partially characterized. Binding of [125I]iodohuman(h)GH was rapid, reversible, and was time and temperature dependent. Maximum specific binding occurred at 37 C in approximately 40 min at pH 7.4. Bound labeled hGH was rapidly dissociable, with the addition of excess unlabeled hormone. Specific binding is inhibited by as little as 1.0-1.5 ng/ml hGH, and 50% inhibition was obtained with 15-20 ng/ml. No inhibition was observed with insulin, glucagon, hPRL, or hTSH at concentrations up to 1 micrograms/ml. This receptor does not discriminate between monkey GH, rat GH, bovine GH, and porcine GH. Specific binding varied linearly with cell concentration. Scatchard analysis revealed linear plots with a Ka of approximately 10(9) M-1 and 15,000 sites per cell. There was less than 15% degradation of [125I]iodo-hGH over 90 min at 37 C. There was a striking increase in [125I]iodo-hGH binding to adipocytes at pH 4.85. Scatchard analysis of binding at pH 4.85 revealed a curvilinear plot with an apparent increase of sites per cell from 15,000 to 60,000, and a modest increase in the apparent affinity constant of the high affinity, low capacity sites using the two-compartment model for curvilinear plots. The GH receptors in rat fat cells displayed no ability to bind labeled hPRL or human placental lactogen, consistent with minimal recognition of lactogenic peptides by these receptors. Thus, the rat adipocyte contains specific binding sites for GH that fulfill the major criteria for receptor binding. The presence of such receptors in these cells may facilitate the study of GH receptors in relation to the biological effects of the hormone on adipose tissue in various metabolic settings. PMID- 6276143 TI - Dopaminergic inhibition of pituitary beta-endorphin-like immunoreactivity secretion in the rat. AB - A pharmacologic approach was used to examine the possible role of dopamine neurons in the regulation of pituitary beta-endorphin-like immunoreactivity (beta END-LI) secretion in the rat. Blockade of dopamine receptors by haloperidol or pimozide treatment evoked dose- and time-related increases in plasma levels of beta-END-LI. Physical immobilization increased circulating beta-END-LI six-fold and pretreatment with dopamine receptor agonists (bromocriptine or pergolide) significantly attenuated this rise without affecting plasma beta-END-LI levels in non-stressed animals. Dopaminergic drugs and immobilization produced similar effects on circulating PRL as on beta-END-LI. However, the magnitude of change in levels of PRL was generally greater than the change in beta-END-LI. The present findings suggest that dopaminergic neurons inhibit the release of pituitary beta END-LI as well as PRL in the rat. PMID- 6276144 TI - The postnatal development of the pituitary thyrotropin-releasing hormone receptor in male and female rats. AB - The density of the pituitary TRH receptor in male and female rats was measured during postnatal development. The younger rats of both sexes exhibited higher [3H]TRH binding than did adults. A comparison of the properties of receptor binding between 4-5 day-old and adult pituitary showed that the increased binding was solely due to an increase in Bmax (maximum binding capacity) and not the KD (binding affinity). In conclusion, these findings demonstrate that the increased TRH-receptor density may be a contributing factor in eliciting exaggerated TSH response to exogenous TRH in neonatal rats. PMID- 6276145 TI - Evidence for the presence and characterization of angiotensin II receptors in rat anterior pituitary membranes. AB - Previous work from this laboratory (1) indicates that angiotensin II (AII) can affect release of several anterior pituitary hormones, both in vivo and in vitro. To ascertain whether specific receptors mediate the effects of AII on the anterior pituitary, specific binding as well as the kinetics of [125I] AII binding to rat anterior pituitary membranes were analyzed. Binding of [125I] AII was rapid, reaching equilibrium within 4 min at 37 C. Specific binding was approximately 90%. Increasing concentrations of ligand resulted in saturation of binding, with equilibrium attained at [125I] AII = 2 nM. Scatchard analysis of the data indicated a single class of binding sites, with an equilibrium dissociation constant, Kd = 0.49 nM, and a maximum binding capacity of 40 fmol/mg protein. Specific binding was directly proportional to membrane protein concentration (range 20-240 micrograms protein). Binding was competitively inhibited on an equimolar basis by (Sar1, Ala8) AII (Saralasin), a specific AII receptor antagonist. The decapeptide Angiotensin I was about 10-20-fold less potent in inhibiting specific AII binding. These studies demonstrate and characterize specific receptor sites for AII in the anterior pituitary gland and offer additional evidence for a role of AII in the regulation of anterior pituitary hormone secretion. PMID- 6276146 TI - Paradoxical effects of cycloheximide on the ultra-rapid induction of two hepatic mRNA sequences by triiodothyronine (T3). AB - Triiodothyronine (T3) rapidly induces the accumulation of two hepatic mRNA sequences (spot 14 and spot CyT) in thyroidectomized rats as revealed by two dimensional gel electrophoresis of in vitro translated products of isolated poly(A) containing RNA. T3 acting alone induced a 29-fold increase of spot 14 within 4 h, an increase which was completely inhibited by the concomitant administration of cycloheximide. On the other hand, CyT could be detected only after the administration of cycloheximide, and the combined action of cycloheximide and T3 resulted in a 4-fold increase in CyT 4 h after administration of both agents. These observations suggested that the early cellular action of T3 is contingent on the participation of rapidly turning over protein and mRNA. PMID- 6276147 TI - Inhibition of lordosis behavior in the female rat by intraventricular infusion of prolactin and by chronic hyperprolactinemia. AB - The role of prolactin (PRL) in the control of the lordosis reflex of female rats was investigated. In the initial series of experiments, the normal high level of sexual receptivity observed in the ovariectomized, estrogen-progesterone (E-P) primed female rat was suppressed by intraventricular infusion of 100 ng PRL. Mating behavior remained suppressed 2, 3, and 5 hours following a single infusion of PRL into the third ventricle. In contrast, infusions of either an equal volume of the solvent vehicle (saline) or 100 ng of adrenocorticotropic hormone (ACTH) were ineffective in modulating the level of mating behavior in hormone-primed female rats. In a second series of experiments, chronic hyperprolactinemia was induced by pituitary transplants under the renal capsule in intact, normal cycle diestrus rats (N=12). A significant decrement in E-P induced mating behavior was observed at 12 and 14 weeks posttransplantation but not at 4 weeks. Sham-operated animals (N=12) displayed the characteristic pattern of behavior normally observed under exogenous E-P therapy. In summary, transient exposure as well as chronic exposure to high levels of PRL can suppress mating behavior, thus suggesting a possible role for PRL in the mediation of reproductive behavior in the female rat. PMID- 6276148 TI - Vasopressin-dependent and -independent control of the release of adrenocorticotropin. AB - We tested the possibility that vasopressin mediates the responses of adrenocorticotropin (ACTH) to electrical stimulation of various areas of the hypothalamus. Thirty-three cats were anesthetized with chloralose-urethane, immobilized with gallamine, and respired artificially. Plasma ACTH was measured by RIA. Intraventricular administration of antiserum to vasopressin blocked the release of ACTH induced by electrical stimulation of the paraventricular nucleus (PVN), suggesting a role for the vasopressinergic projection from PVN to the external zone of the median eminence. In contrast, the release of ACTH induced by stimulation of areas ventral to PVN was unaffected by the antiserum. Thus, there is at least one corticotropin releasing factor released from nuclei other than PVN that is distinct from vasopressin. PMID- 6276149 TI - Water-soluble gonadotropin receptors of the rat ovary. AB - Soluble receptors for LH/hCG were characterized in aqueous and low ionic strength buffer extracts of the luteinized rat ovary. These receptors are proteins with high affinity (KA = 10(10) M-1) and specificity for hCG- and LH-like gonadotropins. The water-soluble receptors are stable and retained 60-80% of their binding capacity after lyophilization. Resolution of the water-soluble receptors by gel electrophoresis demonstrated five binding species, with molecular weights of 165,000, 81,000, 48,000, 24,000 and 12,000. These findings have demonstrated that a proportion (5-10%) of ovarian LH/hCG membrane receptors can be rendered water soluble with preservation of their binding properties and that the specific binding site for LH/hCG is present in proteins much smaller than the predominant 6.5S (mol wt, 194,000) form extracted by nonionic detergents. The stability of these small LH-binding sites is of value for further purification and analysis of structural determinants for gonadotropin binding. PMID- 6276150 TI - X-linked hypophosphatemic mice are not hypersensitive to parathyroid hormone. AB - One of the hypotheses attempting to explain the etiology of the human disease X linked hypophosphatemia (XLH) posits renal hypersensitivity to parathyroid hormone (PTH). These studies were designed to test this hypothesis in vivo, using the hemizygous hypophosphatemic (Hyp/Y) mouse as an animal model for XLH. Vehicle or 1.0 U bovine PTH (bPTH)/g BW sc was given to intact normal or Hyp mice. Two hours later a small but significant hypercalcemia was observed in both genotypes. Only normal mice remained hypercalcemic 5 h after injection. Intact normal mice, but not Hyp mice, displayed a significant bPTH-induced hypophosphatemia. Administration of bPTH caused a significant increase in both fractional excretion of phosphate (FE-P) and urinary cAMP (UcAMP) 2 h after injection. However, there was no significant differences in the magnitude of response between genotypes. In a different experiment hPTH dose-response curves (0, 0.04, 0.2, and 1.0 U bPTH/g BW sc) were constructed in normal and Hyp mice 18 h after thyroparathyroidectomy (TPTX). bPTH caused a significant hypercalcemia and hypophosphatemia in TPTX normal mice at all doses 2 h after injection. But only the highest dose of hormone caused a significant hypercalcemia in TPTX Hyp mice, and no dose caused a significant decrease in plasma P. In both genotypes a dose-dependent increase in FE-P and UcAMP was observed 2 h after bPTH administration. As with intact mice, there was no indication of a hypersensitive or exaggerated renal response in TPTX Hyp mice. In summary, results from these in vivo experiments indicate that the kidneys of Hyp mice are not hypersensitive to exogenous bPTH. Furthermore, TPTX Hyp mice appeared to exhibit "skeletal resistance" to exogenous PTH, as do osteomalacic dogs and humans. We conclude that renal hypersensitivity to PTH does not play a role in the etiology of XLH in Hyp mice. PMID- 6276151 TI - Binding and activation properties of angiotensin II in dispersed rat anterior pituitary cells. AB - The properties of angiotensin II receptors were studied in isolated rat anterior pituitary cells prepared by trypsin digestion. Angiotensin II bound in a time- and temperature-dependent manner to pituitary cells, with Kd of 4.1 x 10(-9) M. The heptapeptide, des-Asp1-angiotensin II, had only one-tenth of the affinity of the octapeptide (Ki = 5.5 x 10(-8) M). These two peptides displayed a similar potency ratio in their ability to stimulate ACTH release from pituitary cells. These results indicate that angiotensin II may play a regulatory role in controlling ACTH secretion from the pituitary gland. PMID- 6276152 TI - Granulosa cell fusion allows heterologous receptor stimulation of adenylate cyclase and progesterone accumulation. AB - The hallmark of the preovulatory follicles in the rat ovary appears to be the presence of receptors for both LH and FSH on follicular granulosa cells. We have tested the possibility that both gonadotropin receptors could share a common adenylate cyclase system utilising cell fusion techniques. Adenylate cyclase (and concomitant stimulation of progesterone synthesis) was inactivated in granulosa cells possessing both FSH and LH receptors by incubation of the cells with N ethylmaleimide. This treatment did not affect hormone binding to the cells. Subsequent fusion, using polyethylene glycol, of the cyclase-inactivated cells with granulosa cells possessing FSH receptors only and an active cyclase restored LH stimulation of cAMP and progesterone production. These findings support the hypothesis that the LH and FSH receptors on granulosa cells of preovulatory follicles share a common adenylate cyclase system. PMID- 6276153 TI - Evidence for an active step in thyroid hormone transport to nuclei: drug inhibition of L-125I-triiodothyronine binding to nuclear receptors in rat pituitary tumor cells. AB - When added to intact GH4Cl or GH3 cells, cytochalasin b, dansylcadaverine, and chloroquine inhibited the binding of L-125I-triiodothyronine (T3) to nuclear receptors within 30 min. These drugs also reduced the amount of 125I-T3 in the post-nuclear supernatant fraction of cells lysed with Triton X-100. If 125I-T3 was added before the drugs, 125I-T3 that was already bound did not dissociate, but further binding was blocked. Inhibition due to 10 microM cytochalasin b or 150 microM dansylcadaverine was reversible within 2 h of drug removal, and inhibition due to 150 microM chloroquine was partially reversible. Drug inhibition was overcome as the T3 concentration was raised. These drugs did not significantly inhibit 125I-T3 binding to isolated nuclei, and nuclei isolated from drug-treated cultures bound as much 125I-T3 as control nuclei. The results suggest that thyroid hormones reach their nuclear binding sites by an active transport process which is inhibited by the drugs studied. PMID- 6276154 TI - Induction of testicular gonadotropin-releasing hormone (GnRH) receptors by GnRH: effects of pituitary hormones and relationship to inhibition of testosterone production. PMID- 6276155 TI - Dependence of pituitary gonadotropin-releasing hormone (GnRH) receptors on GnRH secretion from the hypothalamus. PMID- 6276156 TI - Correlation between prolactin-receptor interaction, down-regulation of receptors, and stimulation of casein and deoxyribonucleic acid biosynthesis in rabbit mammary gland explants. AB - Increasing concentrations of PRL were added to culture media of rabbit mammary gland explants. After 24 h of culture, free and total PRL receptors, casein synthesis, casein messenger (m) RNA concentrations and DNA synthesis were estimated. In the PRL concentration range of 0-100 ng/ml, receptors were progressively down-regulated by the hormone, but no occupied desaturable receptors could be detected. At concentrations between 1,000 and 20,000 ng/ml, the PRL receptors were increasingly occupied without being quantitatively down regulated. There exists a reciprocal correlation between the rate of casein synthesis, casein mRNA concentration, DNA synthesis and the down-regulation of the receptors as a function of PRL concentration. The maximal responses were reached around a PRL concentration of 100 ng/ml. Interestingly, with a large excess of PRL a desensitization process was observed, the responses to the hormone being attenuated. This desensitization was more pronounced for DNA synthesis than for casein synthesis or for casein mRNA accumulation. The data suggest that down-regulation of PRL receptors occurs even at physiological concentrations of PRL. These observations are compatible with the hypothesis that each receptor can generate a limited number of intracellular relays eliciting hormonal action before being irreversibly inactivated and degraded. In addition, high concentrations of PRL induce a refractory state in the mammary gland that is possibly related to a low level of peripheral PRL receptors. PMID- 6276157 TI - Differential effects of pharmacological manipulations of central alpha 1- and alpha 2-adrenergic receptors on the secretion of thyrotropin and growth hormone in male rats. PMID- 6276158 TI - Recovery from thyroid-stimulating hormone-induced refractoriness in thyroid slices: effect of removal of hormone and new protein synthesis. AB - An initial incubation of bovine thyroid slices with TSH causes decreased responsiveness to the subsequent addition of the hormone when the adenylate cyclase -cAMP system and other metabolic parameters are measured. After the initial incubation with TSH, refractoriness persists despite incubation of thyroid slices for 24 h in the absence of added TSH. Removal of persistently bound TSH by trypsin or antibody to TSH did not reverse the refractoriness during a subsequent 2 h incubation without added TSH. However, normal TSH responsivity was restored by the removal of TSH bound during the first incubation by the addition of either trypsin or antibody to TSH at the beginning of a 24-h second incubation. Restitution of TSH responsiveness after treatment with trypsin or antibody to TSH requires new protein synthesis. While TSH-induced refractoriness does not modify stimulation of cAMP by cholera toxin, its effect on glucose oxidation is significantly diminished. Menadiol stimulation of glucose oxidation is not inhibited in thyroid slices refractory to TSH. Thus, the effect of menadiol is subsequent to the block induced by TSH, whereas that of cholera toxin is proximal to it. PMID- 6276159 TI - Immunoreactive gamma-melanotropin in rat pituitary and plasma: a partial characterization. AB - A RIA for the gamma MSH region of proopiomelanocortin has been established and validated. The antiserum was raised to synthetic bovine gamma MSH, and it cross reacts well with larger polypeptides, including gamma MSH and 16K fragment, which contain the gamma MSH sequence. Gel filtration chromatography of extracts prepared from the anterior and neurointermediate lobes of rat pituitary and from rat plasma reveal two heterogeneous immunoreactive (IR-) forms of gamma MSH in each venue which elute in molecular weight peaks of approximately 11,000 (11K) and 6,000 (6K). Addition experiments indicate that the smaller material is not an artifact generated from the larger form during preparation. By the criterion of retention on affinity columns of Concanavalin A-agarose, these peptides are glycosylated. The 6K form represents 8-17% of the total IR-gamma MSH in the anterior lobe and about 30% of that in the neurointermediate lobe. IR-gamma MSH are released by dispersed rat pituitary cells in response to several of the same secretagogues which modulate the secretion of other proopiomelanocortin-derived peptides. Changes in the plasma and anterior lobe content of IR-gamma MSHs and ACTH resulting from perturbation of the pituitary-adrenal axis are concordant. In particular, the plasma concentration of 6K IR-gamma MSH increases dramatically after imposed stress. PMID- 6276161 TI - Identification of the ovulatory follicle in the ewe: associated changes in follicular size, thecal and granulosa cell luteinizing hormone receptors, antral fluid steroids, and circulating hormones during the preovulatory period. PMID- 6276160 TI - Inhibition of granulosa cell differentiation by gonadotropin-releasing hormone. PMID- 6276162 TI - Characterization of an early inhibitory effect of glucocorticoids on stimulated adrenocorticotropin and endorphin release from a clonal strain of mouse pituitary cells. PMID- 6276163 TI - Regulation of gonadotropin receptors and steroidogenesis in cultured porcine Leydig cells. PMID- 6276164 TI - Proopiolipomelanocortin peptides in normal pituitary, pituitary tumor, and plasma of normal and Cushing's horses. AB - Using RIAs for six regions within proopiolipomelanocortin (proOLMC), gel filtration, and electrophoresis, we studied pituitary peptides in a normal horse and one with Cushing's disease caused by a pars intermedia adenoma. Almost all immunoreactive (IR) ACTH (78%) was 4,500 mol wt (4.5K) ACTH in normal pars distalis, but it was almost 100% corticotropin-like intermediate lobe peptide (CLIP) in normal pars intermedia. alpha MSH and beta MSH were found mainly in pars intermedia: equal concentrations of the beta MSH precursors, beta-lipotropin (beta LPH) and gamma LPH, were found in pars distalis. Most IR-beta-endorphin (IR beta END) was found as beta END in pars intermedia, but roughly equal concentrations of beta END and its precursor, beta LPH, were found in pars distalis. A 33K molecule containing IR-ACTH, IR-gamma 3MSH, and IR-beta END, presumed to be proOLMC, and a variety of 15-27K presumed biosynthetic intermediates were found in both normal pars distalis and pars intermedia. The pars intermedia adenoma causing Cushing's syndrome contained high IR-peptide concentrations. Several differences in precursors were noted, including the presence of three larger presumed precursors (38.5K, 47K, and 63K) that had both ACTH and beta END immunoreactivities and both deletions and additions of 15-27K intermediates. The Cushing's horse's plasma peptides reflected tumor concentrations; 4.5K ACTH was modestly elevated, but the concentrations of CLIP, alpha MSH, beta MSH, gamma LPH, and beta END were dramatically increased. About 20% of plasma IR-ACTH and 5% of IR-beta MSH and IR-beta END were found as high molecular weight forms. Normal processing of horse proOLMC appears to be similar to that in other species, but may be altered in pars intermedia tumors of horses with Cushing's disease, the plasma of which contains disproportionately increased concentrations of pars intermedia proOLMC peptides. PMID- 6276165 TI - Decreased in vitro testosterone production by isolated Leydig cells from uremic rats. PMID- 6276166 TI - Interactions of multiplication-stimulating activity with bovine endothelium: comparative studies in primary, passaged, and cloned cultures from the pulmonary and systemic circulations. AB - Endothelial cells were prepared from adult bovine pulmonary and systemic vessels and maintained as primary cultures, passaged cultures, and cloned cell strains. All cultures were shown to contain endothelial cells on the basis of several endothelial-specific and endothelial-associated traits. Both primary and passaged cells demonstrated specific receptors for the insulin-like growth factor, multiplication-stimulating activity (MSA). [125I]Iodo-MSA binding was greatest in cells derived from aortas, least in pulmonary venous cells, and intermediate in pulmonary arterial cells. When compared to MSA receptors on primary endothelial cell cultures derived from human umbilical veins or arteries, the bovine cells bound 4-10 times more MSA per cell. In all bovine cells, insulin competed weakly or not at all with [125I]iodo-MSA binding. The presence of MSA receptors on primary and passaged cells from five different vascular sources (three bovine and two human) suggests that receptors for the insulin-like growth factors may be an intrinsic component of the vascular endothelium. PMID- 6276167 TI - In vitro induction of segregational errors of chromosomes by natural cannabinoids in normal human lymphocytes. AB - It has been shown that segregational errors (SE) of chromosomes can be induced by olivetol and several halogenated inhalation anesthetics. The purpose of this study was to examine the effects of natural cannabinoids - including delta-9 tetrahydrocannabinol (THC), cannabinol (CBN), and cannabidiol (CBD) - on chromosome segregation. Lymphocytes obtained from healthy adult males were incubated with various concentrations of natural cannabinoids for 72 hours. Anaphase preparations were made from these cultures. A statistically significant increase in the incidence of SE of chromosomes was observed in the lymphocytes exposed to THC at a concentration of 3.2 x 10(-6) M, but not to CBN or CBD. A greater incidence of bridge formations, anaphase lags, micronuclei, and unequal segregations in bipolar divisions and multipolar divisions were observed in THC treated lymphocytes, compared with the controls. However, only anaphase lags and unequal segregations in bipolar divisions reached statistically significant levels. It appears that THC affects the formation of microtubules and spindles and may be considered as a mitotic poison. The value of examining SE as a part of cytogenetic studies on chemical mutagens is emphasized. A description of a classification system of SE developed in our laboratory is also presented. This system can be applied to studies using both normal human lymphocyte cultures and lymphoid cell lines. PMID- 6276168 TI - Evidence for a Mg2+-induced conformational change at the ATP-binding site of (Na+ + K+)-ATPase demonstrated with a photoreactive ATP-analogue. AB - 1. The 3'-ribosyl ester of ATP with 2-nitro-4-azidophenyl propionic acid has been prepared and its ability to act as a photoaffinity label of (Na+ + K+)-ATPase has been tested. 2. In the dark 3'-O-[3-(2-nitro-4-azidophenyl)-propionyl]adenosine triphosphate (N3-ATP) is a substrate of (Na+ + K+)-ATPase and a competitive inhibitor of ATP hydrolysis. 3. Upon irradiation by ultraviolet light, N3-ATP photolabels the high-affinity ATP-binding site and is covalently attached to the alpha-subunit and an approximately 12000-Mr component. 4. Photolabeling of the alpha-subunit by N3-ATP irreversibly inactivates (Na+ + K+)-ATPase. 5. Photoinactivation is strictly Mg2+-dependent. Na+ enhances the inactivation. ATP or ADP and K+ protect the enzyme against inactivation. 6. Mg2+, in concentrations required for photoinactivation, protects (Na+ + K+)-ATPase against inactivation by tryptic digestion under controlled conditions. 7. It is assumed that a conformational change of the ATP-binding site of (Na+ + K+)-ATPase occurs upon binding of Mg2+ to a low-affinity site. PMID- 6276169 TI - Effects of cations on high-affinity and low-affinity ATP-binding sites of (Na+, K+)-ATPase as studied by disulfides of thioinosine triphosphate and its analogue. PMID- 6276170 TI - Assembly of cytochrome c. Apocytochrome c is bound to specific sites on mitochondria before its conversion to holocytochrome c. PMID- 6276171 TI - Larger precursors of mitochondrial translation products in Neurospora crassa. Indications for a precursor of subunit 1 of cytochrome c oxidase. AB - Specific labeling in vivo of the formylated N termini of mitochondrial translation products revealed that some mitochondrially synthesized proteins were not labeled this way. As a consequence, it was worthwhile considering that larger precursor proteins of mitochondrial translation products exist. Although we used a rapid isolation procedure, only after 2-h of labeling in the presence of cycloheximide, could three additional mitochondrial translation products (molecular mass 45, 36, and 25 kilodaltons) be detected. Preincubation with cycloheximide indicated that these proteins might be larger precursors which were no longer processed due to the prolonged presence of cycloheximide. To prevent processing of the precursors during isolation, cells of the slime mutant were directly lysed in boiling sodium dodecyl sulphate solution. In this way, the same three additional mitochondrial translation products were detected after a pulse labeling of 1 min. These proteins behave in a precursor-like fashion. Labeling at 9 degrees C resulted in a partial accumulation of the three additional proteins. Finally protein blots treated with antibodies and 125I-labeled protein A, support the idea that the 45-kDa protein is a precursor of subunit 1 of cytochrome c oxidase; 50-80% of this precursor could be detected in the post-mitochondrial supernatant, indicating that this polypeptide is not tightly bound to the membrane. PMID- 6276172 TI - The isolation, mapping and transcription in vitro of a beta 0-thalassaemia globin gene. AB - The red blood cell precursors of a patient with homozygous beta 0-thalassaemia have previously been shown to contain nuclear, but not cytoplasmic, beta-globin specific transcripts. We describe the isolation of a beta-globin gene from this patient as a recombinant bacteriophage chromosome. Restriction-enzyme cleavage site mapping experiments demonstrate no detectable deletions, insertions or major rearrangements in this thalassaemia gene. Two different techniques show that the gene isolated is transcribed as efficiently in vitro as the normal beta-globin gene. PMID- 6276173 TI - Mossbauer studies on the active Fe ... [2Fe-2S] site of putidamonooxin, its electron transport and dioxygen activation mechanism. AB - Putidamonooxin, the oxygenase of a 4-methoxybenzoate monooxygenase enzyme system, catalyzes the oxidative O-demethylation of the substrate 4-methoxybenzoate in conjunction with the NADH:putidamonooxin oxidoreductase. Putidamonooxin is a conjugated iron-sulfur protein which needs iron ions as cofactors for its enzymatic activity. Putiamonooxin was isolated from Pseudomonas putida, which was grown on a 57Fe-enriched culture medium. Thus putidamonooxin was enriched in vivo with 57Fe up to about 80%. During our Mossbauer study of putidamonooxin a number of parameters have been varied: (a) the oxidation state of putidamonooxin (oxidized, reduced and aerobically reoxidized); (b) the substrate bound to putidamonooxin (4-methoxybenzoate, benzoate, 4-tert-butylbenzoate); (c) the temperature between 2.7 K and 245 K; (d) the applied magnetic field between 0 and 0.1 T and (e) the amount of iron cofactor. From our Mossbauer results it is obvious that the iron-sulfur centers of putidamonooxin are [2 Fe-2S] clusters similar to those of the plant-type ferredoxins. Further, we have evidence for the existence of iron ions (one per [2 Fe-2S] cluster), which serve as cofactors for the dioxygen activation, functioning as the dioxygen binding site and mediating the electron flow from the [2 Fe-2S] cluster to dioxygen. PMID- 6276174 TI - The effect of the redox potential on the activity of the nitrogenase and on the Fe-protein of Azotobacter vinelandii. PMID- 6276175 TI - Comparison of the effects of oligomycin and dicyclohexylcarbodiimide on mitochondrial ATPase and related reactions. PMID- 6276176 TI - Dynamics of erabutoxin b as studied by nuclear magnetic resonance. Relaxation studies of methyl proton resonances. AB - Longitudinal and transverse relaxation times were measured for well-resolved and assigned methyl proton resonances of erabutoxin b at 270 MHz, 300 MHz and 500 MHz. Both longitudinal and transverse magnetization decay curves are non exponential due to cross-relaxation and cross-correlation effects. The longitudinal and transverse relaxation rates were obtained from the initial slope of both magnetization decay curves. The correlation times for the isotropic tumbling motion of the protein were determined to be 2.82 ns at 300 K and 1.62 ns at 330 K from the analysis of the relaxation data of some alpha protons. Using these values, the relaxation data of methyl protons were fitted to various theoretical models. Most of the methyl resonances could be fitted well to a model which allowed methyl rotation (in the range 0.01-0.05 ns) and an external contribution from protons assumed to be in positions derived from X-ray coordinates. The data for a few methyl groups, however, could not be fitted in this way. For these a smaller number of external protons than predicted by the X ray coordinates was assumed. Additionally, a larger amplitude motion had to be introduced into the model for particular residues. This additional motion requires concerted protein motion close to these residues, since the X-ray structure suggests that steric hindrance would prevent local motion. These results are consistent with the idea of a flexible and dynamic structure for proteins. PMID- 6276177 TI - Collagenase in the Walker 256 carcinoma. A study of the latent and active enzyme in vivo and in vitro. AB - A latent form of collagenase had been isolated from crude extracts of the insoluble, fibrous material from Walker tumor homogenates. Purified preparations of this enzyme yielded a major unit of Mr approximately 62000, as determined by gel filtration on AcA 54 Ultrogel. In its activated form collagenase had been purified to apparent homogeneity with an approximate Mr of 42000. The active enzyme cleaved soluble collagen into three-quarter and one-quarter length fragments in the manner of vertebrate collagenases. Latent collagenase from culture media eluted with an apparent Mr of 53000 and was thus slightly larger in size than its activated form that eluted at 42000. Extracted latent collagenase and latent collagenase from culture media could be activated enzymatically by trypsin or chymotrypsin and non-enzymatically by mersalyl, an organic mercurial compound. We suggest that latent collagenase from Walker tumors are complexes of active enzyme with inhibitor(s) of low molecular weight(s) and are not true zymogens. PMID- 6276178 TI - Kinetic studies of proton transfer in the microenvironment of a binding site. AB - Excitation of 8-hydroxypyrene 1,3,6-trisulfonate to its first electronic singlet state converts the compound from weak base (pK degrees = 7.7) into a strong acid (pK* = 0.5). The dissociation of the proton in water or dilute salt solution is a very fast reaction, K12 = 1 X 10(10) S-1. In concentrated salt solutions the dissociation is slowed as an exponential function of the chemical activity of the water in the solution. This kinetic parameter has been used to gauge the properties of the microenvironment of the binding sites of bovine serum albumin at which this compound is bound. Time-resolved fluorometry reveals two distinct steps: a rapid dissociation of the proton with tau = 300 +/- 40 ps which lasts approximately 0.5 ns, followed by a slower reaction with tau = 3.3 ns. The first rapid phase represents proton dissociation taking place in the binding site. From the rate constant K = 3.3 X 10(9) s-1 we estimate that the ability of the water molecules in the site to hydrate the ejected proton is equivalent to a salt solution with water activity of 0.85. The slow phase represents the escape of the proton from the binding site. The rate of the escape, 1.4 X 10(8) s-1, is significantly slower than diffusion-controlled dissociation. It is concluded that the shape of the site or its lowered proton conductivity do not allow a rapid escape of the proton to the bulk. Still it should be remembered that the escape of the proton is 10(5)-10(6)-times faster than a typical turnover of an enzyme.U PMID- 6276179 TI - Direct evidence of a glucagon-dependent regulation of the concentration of glucagon receptors in the liver. PMID- 6276180 TI - Evidence for intragranular processing of pro-opiocortin in the mouse pituitary intermediate lobe. AB - Mouse pituitary neurointermediate lobes were pulse-incubated in [3H] arginine or [3H] lysine for 10 min and then chase-incubated for periods 0 to 4h. The labeled peptides from the lobes were analysed by immunoprecipitation with specific antisera, and thereafter, by acid-urea polyacrylamide gel electrophoresis. Using this paradigm, the synthesis of a prohormone common to adrenocorticotropin (ACTH) and endorphin was detected in 10 min pulse labeled lobes. Following a chase period, processing of the prohormone to several forms of ACTH (mol. wt. 25000, 23000, and 13000), beta-lipotropin and beta-endorphin was observed. To determine the intracellular site of processing of the prohormone, subcellular fractionation studies of labeled lobes were carried out. Analysis of the fractions from the pulse-labeled lobes revealed that the newly synthesized labeled prohormone was primarily localized in a granule-enriched fraction. In lobes that were pulsed and then chase-incubated for 1 h, there was a decrease in the amount of prohormone and an appearance of processed products in the granule-enriched fraction. In another paradigm, where the secretory granule-fraction was isolated from pulse labeled lobes and then incubated in vitro for 6 h at pH 5.5, processing of the endogenous labeled prohormone within the isolated granule fraction was observed. These data suggest, that in the mouse neurointermediate lobe, the ACTH/endorphin prohormone (pro-opiocortin) is rapidly packaged into secretory granules after synthesis and processed intragranularly. PMID- 6276181 TI - Plasma membrane specialization and intracellular polarity of freshly isolated rat hepatocytes. AB - It was investigated whether rat hepatocytes maintain their plasma membrane specialization (sinusoidal, lateral and bile canalicular sites) and their intracellular polarity (peribiliary region, rich in lysosomes and poor in mitochondria) after isolation. The morphology of the hepatocytes and the cytochemical localization of marker enzymes for the bile canalicular membrane (alkaline phosphatase, adenosine triphosphatase and 5' nucleotidase), for the lysosomes (acid phosphatase) and for the mitochondria (beta-hydroxybutyrate dehydrogenase and succinate dehydrogenase) were studied in situ and directly after isolation using both light and electron microscopy. The morphology of the cells and the cytochemical activity of acid phosphatase, succinate dehydrogenase and beta-hydroxybutyrate dehydrogenase showed that in isolated cells, as in situ, the lysosomes were concentrated in bands, devoid of mitochondria. Unlike in situ the reaction product of alkaline phosphatase, adenosine triphosphatase and 5'nucleotidase was evenly distributed along the entire plasma membrane of the isolated cells. Morphologically, no tight or gap junctions or desmosomes could be detected in the isolated cells, while the plasma membrane appeared to be homogeneously covered with uniform microvilli. In conclusion it can be stated that during isolation the hepatocytes loose their distinct plasma membrane specialization, but maintain their peribiliary region rich in lysosomes and poor in mitochondria. PMID- 6276182 TI - Expression of the murine mammary tumor virus in the milk of F1 hybrids of virus negative mouse strains. PMID- 6276183 TI - Growth kinetics of small cell carcinoma of the lung. PMID- 6276184 TI - Effects of megestrol acetate on growth and secretion of a pituitary tumor. PMID- 6276185 TI - Chemotherapy of small cell carcinoma of the lung: comparison of a cyclic alternative combination with simultaneous combinations of four and seven agents. PMID- 6276186 TI - Polymorphism of germ-line immunoglobulin VH genes correlates with allotype and idiotype markers. AB - The polymorphic nature of the immunoglobulin VH genes was investigated by Southern blot analysis of liver DNA of sixteen different mouse strains and hybridization with VH probes. Differences in restriction enzyme pattern (REP) were observed and six different patterns of restriction fragments were found for the sixteen strains analyzed. No equivalent polymorphism was observed in another multigene family, the actins. The six patterns correlate with immunoglobin constant region allotypes (Igh-1). Experiments with Igh-1-congenic strains suggest that the VH REP is linked to immunoglobulin constant region haplotype. Mouse strains which share inherited idiotypes also share identical VH restriction pattern. This provides a structural basis for the genetic linkage between idiotypes and allotypes. It also indicates that different strains carry different VH gene repertoires, which may be the basis for the expression of different inherited idiotypes in various strains. We propose that a VH group in a set of linked genes that are coinherited as a cluster with the constant region genes and that VH and Ch can be regarded as an extended haplotype. PMID- 6276187 TI - Spinal and Supraspinal sites for morphine and nefopam analgesia in the mouse. AB - Using the tail-flick and hot-plate assays, morphine and nefopam were tested for analgesic activity following intraperitoneal (i.p.), intracranial (i.c.) and intraspinal (i.s.) injection in mice. By the i.p. route, morphine was equipotent on both analgesic tests. Nefopam was one-third as potent as morphine on the hot plate test, but did not affect the tail-flick. Intracranial morphine was more effective on the hot-plate than on the tail-flick, but i.s. morphine was most potent on the tail-flick. Naloxone, 0.5 mg/kg i.p., totally reversed morphine's effects on the tail-flick, but only partially reversed these actions on the hot plate, suggesting the possibility that morphine's effects on the mouse hot-plate test may be mediated via multiple receptor types. Nefopam was more potent by the i.c. route than by the i.p. route, but its was inactive spinally. Nefopam analgesia was unaffected by naloxone treatment. It is concluded that nefopam is a novel, centrally acting, non-narcotic analgesic. PMID- 6276188 TI - Intraventricular cholecystokinin-octapeptide produces hypothermia in rats. PMID- 6276189 TI - Presynaptic inhibition of sympathetic neurotransmission by adenosine in the rat kidney. AB - The effect of adenosine on responses to sympathetic nerve stimulation was studied in the isolated perfused rat kidney. Adenosine at 1, 3 and 10 micrograms/ml caused significant impairment of renal sympathetic neurotransmission as evidenced in a dose-dependent reduction in the vasoconstrictor responses elicited by periarterial nerve stimulation at 2, 4 and 8 Hz. Vasoconstriction to exogenous norepinephrine was unaffected by adenosine except at 10 micrograms/ml where there was a slight reduction in response to norepinephrine. Theophylline, an adenosine receptor antagonist, inhibited the above action of adenosine at the frequencies of 2 and 4 Hz. At 2 Hz, 10 microM theophylline completely blocked the effect of 1 and 3 micrograms/ml adenosine and reduced by 50% the inhibitory action of 10 micrograms/ml. Higher concentration of theophylline (50 microM) was required to antagonize the action of adenosine at 4 Hz. These results indicate that adenosine can inhibit sympathetic neurotransmission in the rat kidney through a presynaptic purinergic mechanism. The physiological and pharmacological significance of this presynaptic action of adenosine is discussed. PMID- 6276190 TI - Cyclic AMP increase in canine parotid gland mediated by histamine H2-receptors. AB - Histamine, 4-methylhistamine and dimaprit induced a dose-dependent increase in the cyclic AMP content of chopped canine parotid gland in the presence of 3 isobutyl-1-methylxanthine. Metiamide, but not mepyramine, inhibited the effect of these agonists dose-dependently. The cyclic AMP content was also increased by 2 (2-pyridyl)ethylamine. This effect was completely blocked by propranolol. These results indicate that the cyclic AMP response to histamine in canine parotid gland is mediated by H2-receptors. Mepyramine enhanced the effect of histamine. However, 2-(2-pyridyl)ethylamine had no significant inhibitory effect on the cyclic AMP response to 4-methylhistamine. A combination of aminoguanidine and quinacrine potentiated the effect of histamine, though no difference was observed in the histamine concentration of the media with or without these inhibitors when determined at the end of incubation. This suggests that there exist active histamine-metabolizing systems in canine parotid gland. PMID- 6276192 TI - Enhancement of cocaine-induced hyperactivity in mice by benzodiazepines: evidence for an interaction of GABAergic processes with catecholaminergic neurons? AB - The effects of nine benzodiazepines on the locomotor stimulation induced in mice by cocaine (4 mg . kg-1 i.p.) were studied. These benzodiazepines markedly enhanced cocaine-induced hyperactivity. This effect was observed at low doses, e.g. doses at least 8 times lower than those required to depress the stimulation caused by cocaine. Nitrazepam-induced enhancement of the hyperactivity elicited by cocaine was reduced or suppressed by blocking dopaminergic receptors with pimozide (0.015--0.03 mg . kg-1), by interrupting GABAergic transmission with picrotoxin (0.25--0.5 mg . kg-1) or blocking alpha- or beta-adrenergic receptors with prazosin (0.25 mg . kg-1) or dl-propranolol (4 mg . kg-1) respectively. At these doses, neither pimozide, picrotoxin, prazosin nor propranolol were able to modify the spontaneous locomotor activity or the stimulation elicited by cocaine alone. Strychnine (0.25--0.50 mg . kg-1) or methysergide (2 mg . kg-1) failed to alter the enhancement by nitrazepam of cocaine-induced hyperactivity. These results suggest that an interaction of benzodiazepines with some catecholaminergic processes, either directly or through the involvement of a GABAergic link, may account for their facilitatory activity on cocaine-induced locomotor stimulation. PMID- 6276191 TI - Influence of age on the effects of chronic fluphenazine on receptor binding in rat brain. AB - The effects of age on alterations in brain dopaminergic (spiperone), beta adrenergic (DHA), alpha-adrenergic (WB-4101), and cholinergic (QNB) binding induced by chronic administration of fluphenazine was studied in the rat. Compared to age-matched saline controls, older-age (25 month) fluphenazine rats showed: (a) a similar percent increase in specific spiperone binding (at 0.1 nM) but a slightly smaller increase in the Bmax of spiperone binding in the striatum than younger fluphenazine-treated rats; (b) a substantially greater increase in the Bmax of DHA binding and a slightly greater increase in Bmax of WB-4101 binding in the cerebral cortex than younger fluphenazine-treated rats. There was no significant interaction of age with the effect of chronic fluphenazine on QNB binding in rat striatum. PMID- 6276193 TI - Clonidine inhibition of pancreatic secretion in rats: a possible central site of action. AB - The effects of clonidine on pancreatic secretion were studied in rats fitted with chronic or acute fistulas. Subcutaneous and intracerebroventricular injections of clonidine in conscious rats induced a dose-dependent inhibition of basal pancreatic secretion involving volume, bicarbonate output and protein output with an ED50 of about 10 micrograms/kg. Clonidine inhibition of pancreatic secretion was not dependent on the associated inhibition of gastric acid output. In conscious rats, the pancreatic inhibitory effect of clonidine was completely antagonized by yohimbine and slightly by piperoxane and prazosin. Propranolol, mianserin, naloxone and cimetidine did not antagonize the clonidine effect. Clonidine decreased the basal pancreatic secretion in anaesthetized rats and this action was completely reversed by yohimbine. Clonidine inhibited the pancreatic secretion stimulated by 2-deoxyglucose. This effect was reversed by yohimbine, while prazosin had no effect. Clonidine did not inhibit the pancreatic secretion induced by electrical stimulation of the vagus nerves. These results suggest that clonidine inhibition of pancreatic secretion is mediated through alpha 2 adrenergic receptors, and at least in part by a central nervous system mechanism. Yohimbine alone increased basal pancreatic secretion in conscious rats. This suggests that alpha 2-adrenergic receptors might be involved in the physiological nerve tone to the pancreas. PMID- 6276194 TI - Calcium-opiate antagonism in the periaqueductal grey (PGA) region. PMID- 6276195 TI - Dose-dependent feeding in the rat following substantia nigra acetylcholinesterase blockade. PMID- 6276196 TI - Potentiation of bovine growth hormone release by an inhibitor of potassium channels. PMID- 6276197 TI - In vivo alpha 2-adrenoceptor-mediated inhibition of isoproterenol-stimulated free fatty acid output in dog bone marrow adipose tissue. PMID- 6276198 TI - Coronary spasm produced by picrotoxin in cats. AB - Picrotoxin (2 mg/kg i.v.) was administered to 6 vagotomized chloralose anesthetized cats while monitoring coronary blood flow (electromagnetic flow probe), blood pressure and ECG. Coronary constriction occurred in each animal as measured by an increase in coronary vascular resistance (CVR) (mean = 25.2 +/- 7.5%, P less than 0.05) followed by ST segment elevation (0.30 +/- 0.06 mV, P less than 0.05). Pretreatment with phentolamine prevented the increase in CVR and the elevation in ST segment. These results demonstrate that picrotoxin produces alpha-receptor mediated coronary spasm. PMID- 6276199 TI - A characterization of dynorphin-(1-13) on the guinea pig ileal longitudinal muscle. AB - Using naloxone as the antagonist, a comparison of pA2 values obtained from the guinea pig ileal longitudinal muscle preparation revealed that the pA2 value for dynorphin-(1-13) was significantly different from that of the pure narcotic agonists such as morphine, beta h-endorphin, Leu- and Met-enkephalin and that of the mixed agonist-antagonists such as nalorphine. In addition, no cross-tolerance to dynorphin-(1-13) could be demonstrated whereas a pronounced cross-tolerance existed for other opioid peptides on a ileal strip made tolerant to morphine by implantation of morphine pellets to the guinea pig for 72 h. Thus, dynorphin-(1 13) would appear to have a unique pharmacology on this peripheral opioid receptor preparation quite distinct from that of other known opioid peptides. PMID- 6276200 TI - [3H]rauwolscine (alpha-yohimbine): a specific antagonist radioligand for brain alpha 2-adrenergic receptors. AB - [3H]Rauwolscine, a specific and potent alpha 2-antagonist radioligand, was used to characterize alpha 2-receptor binding in bovine cerebral cortex. [3H]Rauwolscine binding was reversible, stereospecific, and saturable. Association, dissociation, and saturation studies revealed one site interactions (k -1/k+1 = 1.2 nM, KD = 2.5 nM, Bmax = 160 fmol/mg protein) and competition studies indicated that [3H]rauwolscine labeled the alpha 2-receptor. Agonists inhibited [3H]rauwolscine binding in a shallow, GTP-sensitive manner. These results suggest that [3H]rauwolscine specifically labels both the high and low affinity states of the alpha 2-receptor in brain membranes. PMID- 6276201 TI - In situ molecular weight determination of brain and peripheral benzodiazepine binding sites. PMID- 6276202 TI - Studies on calmodulin isolated from Tetrahymena cilia and its localization within the cilium. PMID- 6276203 TI - RNA polymerase II in C6 glioma cells. Alpha-amanitin blockade of cAMP phosphodiesterase induction by beta-adrenergic stimulation. PMID- 6276204 TI - Expression of transformation-associated traits in the myogenic cell lines L6 and L8. PMID- 6276205 TI - Digitalis-induced stimulation. Support for a two-receptor model. PMID- 6276207 TI - The induction of spore cells in vitro by membranes of Dictyostelium discoideum. PMID- 6276206 TI - Thyrotropin induces changes in the morphology and the organization of microfilament structures in cultured thyroid cells. PMID- 6276208 TI - Binding of ligands to the asialoglycoprotein receptor causes a reduction in Ca2+ flux through the plasma membrane of isolated hepatocytes. PMID- 6276209 TI - Localization of genes on fractionated rat chromosomes by molecular hybridization. PMID- 6276210 TI - Identification of beta-adrenergic receptors in the pigmented mammalian iris ciliary body diaphragm. PMID- 6276211 TI - Normal and conditionally transformed bovine lens epithelial cell lines containing alpha-and gamma-crystallin. PMID- 6276212 TI - Beta 2 receptor-mediated stimulation of Friend erythroleukemia cell growth by thyroid hormones. AB - Thyroid hormones are known to enhance normal erythroid colony growth (CFUE) and this enhancement depends on a functional beta 2-adrenergic receptor mechanism. we investigated the response of Friend cells to thyroid hormones, catecholamines, and other compounds influencing cellular cAMP activity. The thyroid hormones L T3, L-T4, and "reverse T3" stimulated erythroleukemia colony growth in a serum substituted methylcellulose culture system with peak activity at 10(-7) M. Various beta-adrenergic compounds enhanced Friend leukemia colony growth; however, the alpha-adrenergic agonist phenylephrine was inactive. Dibutyryl cyclic AMP and the phosphodiesterase inhibitor theophylline also enhanced Friend leukemia colony formation. Adrenergic antagonists with beta 2 specificity abrogated the stimulatory effect of L-T3, L-T4, and of "reverse T3" at equimolar concentrations. These experiments demonstrate that thyroid hormones, beta adrenergic agonists, the phosphodiesterase inhibitor theophylline, and dbcAMP have a direct effect on the proliferation of Friend erythroleukemia cells. We conclude that thyroid hormones' action requires a functioning beta 2-adrenergic receptor mechanism. Thyroid hormones directly modulate the growth of neoplastic erythroid cells in a manner consistent with their effects on normal erythropoiesis. PMID- 6276214 TI - DNA-polymerase induced by Herpesvirus papio (HVP) in cells of lymphoblastoid cultures derived from lymphomatous baboons. Report V. AB - A new DNA-polymerase was found in the cells of suspension lymphoblastoid cultures which produce lymphotropic baboon herpesvirus (HVP). This enzyme was isolated in a partially purified form. Some of its properties vary from those of other cellular DNA-polymerases. HVP-induced DNA-polymerase has a molecule weight of 160,000 and sedimentation coefficient of about 8 S. The enzyme is resistant to high salt concentration and N-ethylmaleimide, but it is very sensitive to phosphonoacetate. It effectively copies "activated" DNA and synthetic deoxyribohomopolymers. Attempts to reveal the DNA-polymerase activity in HVP virions were unsuccessful. PMID- 6276213 TI - Schistosoma mansoni: rapid isolation and purification of schistosomula of different developmental stages by centrifugation on discontinuous density gradients of Percoll. PMID- 6276215 TI - The effect of inorganic pyrophosphate (PPi) anions on the in vitro collagen fibril formation. AB - This paper presents a study on the effect of sodium salt of pyrophosphate (Na PPi) in different concentrations (1-10 mMol/l) on the in vitro collagen fibril formation. Collagen was mixture of type I and III collagen dissolved in neutral buffer containing sodium chloride of 1.0 mol/l. Na-PPi added to collagen delayed the fibrillary precipitation of collagen from the solution. Electronmicroscopically the fibrils are of native collagen type, but they are thicker and show a pattern of fine substriation. Calcium and other divalent cations showed differences in their influence on the effect of Na-PPi. This experimental model was developed in order to interpret the supposed role of PPi in the inhibition of apatite crystal formations as well as the fibrogenic effect of its calcium salt in PPi-arthropathy. The effect of PPi on the collagen fibril formation should also be taken into consideration. PMID- 6276216 TI - Demonstration of injured hepatocytes after carbon tetrachloride administration by loss of histochemical glucose-6-phosphatase reaction. PMID- 6276217 TI - Electroncytochemical studies on changes of the adenylcyclase activity following experimental hypoxia and ischaemia in perfused rat hearts. AB - Changes in activity and localization of adenylcyclase were studied in rat heart perfused according to LANGENDORFF after N2-and CO-hypoxia and ischaemia. Under hypoxic conditions an increase of adenylcylase activity was found in the first minutes. Following 3 h nitrogen or 30 min CP perfusion the intensity of reaction was diminished. This later decrease of reaction intensity was not encountered under ischaemic conditions. Changes in localization were not observed either in hypoxia or in ischaemia. After reserpine or propranolol pre-treatment the posthypoxic/ischaemic initial activity-increase did not occur. It is assumed that the early increase in adenylcyclase activity (reaction intensity) following hypoxia/ischaemia is due to endogenous catecholamine release. PMID- 6276218 TI - Effects of hypochlorite on thiamine and its derivatives. AB - Thiamine and its phosphorylated derivatives reacted with hypochlorite with reaction rates following the order: thiamine greater than thiamine monophosphate greater than thiamine diphosphate from pH 4.0 to 6.5. At least one unknown transient intermediate was formed and at least one non-thiochrome product was fluorescent. Chemiluminescence was also observed. PMID- 6276219 TI - thiD locus of Escherichia coli. PMID- 6276220 TI - 3H-Diazepam binding sites in Roman high- and low-avoidance rats. AB - The binding of 3H-diazepam to membrane benzodiazepine receptors was examined in 2 psychogenetically selected lines of rats, which differ according to the selection criterion in avoidance behaviour (RHA/Verh greater than RLA/Verh) and, in addition, in emotionality (RHA/Verh less than RLA/Verh). RHA/Verh rats tended to show higher specific diazepam binding in all CNS-subregions when compared with RLA/Verh animals. Significant differences were found in the cortex, striatum, hippocampus, thalamic region and pons-medulla. These results reinforce the contention that a system involving benzodiazepine receptors may play a role in emotional behaviour. PMID- 6276222 TI - Lack of effect of cortisone, thyroxine and insulin on the developmental pattern of mouse intestinal glucose-6-phosphatase activity. PMID- 6276221 TI - Effect of lithium on normal and chronic granulocytic leukemia colony forming cells (CFU-GM). AB - The effect of lithium on normal and chronic granulocytic leukemia (CGL) granulo monocytic precursors (CFU-GM) has been studied. Lithium slightly increases normal CFU-GM growth whereas it is without effect or moderately inhibitory for CGL CFU GM. It is suggested that it is unlikely that lithium therapy enhances the proliferation of a silent leukemic clone. PMID- 6276223 TI - Effect of benzo[a]pyrene on Friend virus leukemogenesis, CFU-S viability, and induction of humoral immunity. PMID- 6276224 TI - Alpha-thioglycerol sensitive excitatory oxytocin receptors in fowl rectum. AB - The selective antagonism of the excitatory responses of oxytocin by alpha thioglycerol and its potentiation by Mg++ indicate the presence of specific oxytocin receptors in fowl rectum. In addition, the sensitivity of the rectum to very small doses of oxytocin suggests a possible facilitatory role of neurohypophyseal hormones in oviposition. PMID- 6276225 TI - The damage of the hepatic mixed functional oxygenase system by CCl4: significance of incorporation of 14CCl4 metabolites in vivo. PMID- 6276226 TI - [Effect of the tetracyclic antidepressants pyrazidol and inkazan on the adrenergic nerve transmission in the rat vas deferens]. AB - The effect of new tetracyclic antidepressants pyrazidol and inkazan on adrenergic neurotransmission in the isolated rat vas deferens was studied by examining vas deferens contractions in response to the transmural electric stimulation of the postganglionic sympathetic nerves and addition of noradrenaline (NA) or BaCl2. Pyrazidol and inkazan were found to be capable to increase vas deferens contractions in response to transmural electric stimulation or addition of NA. When given in high concentrations, these antidepressants inhibit the adrenergic neurotransmission since inkazan possesses low sympatholytic activity and pyrazidol exhibits certain alpha-adrenolytic effect. PMID- 6276227 TI - [Quinolizidinyl derivatives of tricyclic systems]. AB - Since the pharmacological screening of several lupinyl- and lu lupinyldene derivatives of three-ring systems showed several interesting activities on the C.N.S., new investigations have been undertaken in order to understand the neurochemical mechanisms underlying such activities (influence on the uptake of choline, norepinephrine and serotonin by isolated rat brain synaptosomes). Thus the series of lupinyl derivatives has been completed with N-lupinyl-2 chloroiminodibenzyl and N-lupinyl-2,3-hexamethyleneindole; moreover, for all the three-ring systems so far considered, the corresponding epi-lupinyl- and epi lupinylidenderivatives have been prepared in order to check the significance of the steric relationships between the quinolizidine ring and the tricyclic systems. The latter compounds differ from those formerly described for the equatorial position (rather than axial) of the methylene or methine group joining the quinolizidine nucleus to the three-ring systems. PMID- 6276228 TI - Ca2+-dependent association between a Ca2+-activated neutral proteinase (CaANP) and its specific inhibitor. PMID- 6276229 TI - Frequent deletion of Bacillus subtilis chromosomal fragment in artificially constructed phage Rho 11phisA+. PMID- 6276230 TI - Calculating the distribution of ventilation-perfusion ratios from inert gas elimination data. AB - It is well known that the major cause of hypoxemia in lung disease is ventilation perfusion (VA/Q) inequality, but it has been extremely difficult to measure the distribution of ventilation-perfusion ratios except in terms of unrealistically simple (albeit useful) models. The multiple inert gas elimination technique provides considerable information concerning the shape, position, and dispersion of the VA/Q distribution, although it cannot precisely define all features of the distribution. Although there are many techniques for obtaining information about the distribution from inert gas elimination data, we have found the most flexible and useful approach to be a multicomponent analysis with enforced smoothing, sometimes known as ridge regression. This presentation describes in some detail the physiological and mathematical principles principles involved in the transformation of inert gas elimination data into a representative distribution of ventilation-perfusion ratios by enforced smoothing techniques. It is important to realize that with this approach and any other approach aimed at estimating the distribution of ventilation-perfusion ratios, the results must be properly interpreted. PMID- 6276232 TI - Use of noninvasive fluorometry and spectrophotometry to study epithelial metabolism and transport. AB - Various examples illustrating the use of spectrophotometry and fluorometry in epithelia are presented. The first example uses the redox level of cytochrome aa3, measured spectrophotometrically as an index of tissue anoxia in cortical tubules and slices from the rabbit kidney. In the second example the redox level is used to measure the kinetics of aerobic energy production during transition to anoxia in the midgut of the tobacco hornworm. In the third application, the redox level of mitochondrial NADH is measured fluorometrically in a cortical tubule suspension from the rabbit kidney. Inhibition of active transport work causes reduction of NAD whereas increased work elicits oxidation of NAD, both occurring as expected from mitochondrial transitions to a lesser or more active state, respectively. Another use of NADH fluorescence is the determination of the relative effectiveness of metabolic substrates to deliver reducing equivalents to the respiratory chain in a particular tissue. Redox changes in mitochondrial NAD may be used to distinguish between primary metabolic and primary transport effects of hormones, drugs, and changes in the state of the organism. Finally, examples are provided of the use of an intracellular pH-sensitive dye and an extracellular calcium-sensitive dye in kidney tubules. PMID- 6276231 TI - Biochemical analysis of triggering signals induced by bridging of IgE receptors. AB - Bridging of IgE receptors on rat mast cell plasma membranes induces phospholipid methylation and a monophasic increase in cyclic AMP. The stimulation of phospholipid methylation in the plasma membrane appears to be intrinsic to the processes leading to Ca2+ influx and histamine release. Evidence was obtained that IgE receptors are closely associated with methyltransferases and adenylate cyclase in the plasma membranes. The activation of one enzyme is regulated by the other. An increase in the cyclic AMP level before receptor bridging suppressed phospholipid methylation. On the other hand, inhibition of phospholipid methylation may affect the initial rise in cyclic AMP. Our experiments also indicated that bridging the receptor activates a membrane-associated proteolytic enzyme. Inasmuch as the inhibition of the enzyme activation results in the suppression of both phospholipid methylation and initial rise in cyclic AMP induced by receptor bridging, the proteolytic enzyme may be involved in the activation of methyltransferases and adenylate cyclase. PMID- 6276233 TI - Molecular events involved in 1,25-dihydroxyvitamin D3 stimulation of intestinal calcium transport. AB - There is a biphasic response of intestinal calcium transport to 1,25 dihydroxyvitamin D3 (1,25-(OH)2-D3). The first or rapid response is by existng mature villus cells, whereas the slow second response is by maturing crypt cells. For both responses, [3H]1,25-(OH)2-D3 localizes in the nucleus before initiating the transport events. This localization is brought about by a specific cytoplasmic receptor, which has a molecular weight of 67,000, is highly specific for 1,25-(OH)2-D3, and has a Kd of 5 X 10(-11) M. Its essentiality for intestinal calcium transport response to 1,25-(OH)2-D3 can be demonstrated in neonatal rat pups. In cultured chick intestinal duodena calcium transport begins to appear within 4 h after the addition of 1,25-(OH)2-D3. The response of this calcium transport system to 1,25-(OH)2-D3 is totally blocked by cycloheximide in a reversible manner. Similarly, it is blocked by actinomycin D in a partially reversible manner. These results make it obvious that the rapid calcium transport response to 1,25-(OH)2-D3 involves nuclear activity and transcription of DNA into functional proteins. The exact nature of the transport proteins remains largely unknown except for the calcium-binding protein originally discovered by Wasserman and colleagues. The transport proteins are believed to operate at the brush border membrane surface to facilitate the transfer of calcium and phosphorus into the absorption cells. PMID- 6276235 TI - Electron microscopic characterization of avian myeloblastosis virus RNA by the non-protein technique of spreading. AB - The quantitative analysis of the behaviour of retroviral RNA (AMV RNA) under the conditions of the non-protein technique of electron microscopic visualization on a mono-molecular film of BAC was performed. This technique resulted in visualization of intact molecules the mean length of which was 12% larger in comparison with molecules spread by the cytochrome c method. The method was found to be extremely sensitive to the surface properties of the supporting membrane which distinctly affect the shape and size of molecules. Arrangement of the surface potential of the supporting foil by means of ethidium bromide led to high reproducibility of RNA molecule stretching and to an increase in their length. The conditions were worked out in which extended linear RNA molecules were visualized, even under gentle denaturation. These conditions represent a suitable approach to the electron microscopic visualization of the protein--AMV RNA complexes. PMID- 6276234 TI - [Effect of complex-forming compounds on pancreatic insulocytes]. PMID- 6276236 TI - [The effect of thyroid hormones on low density lipoprotein receptors in cultured human skin fibroblasts (author's transl)]. AB - The catabolic rate of low density lipoprotein (LDL) has been shown to be delayed in hypothyroidism and increased in hyperthyroidism. LDL catabolism is mainly mediated through the LDL receptor pathway in the parenchymal cells. In the present paper the effect of thyroid hormones in LDL receptors was studied in cultured human skin fibroblasts. Human fibroblasts were established from 2 normal subjects, one heterozygous patient and 2 homozygous patients with familial hypercholesterolemia (FH). Human LDL (density 1.020 approximately 1.050 g/ml) from the serum of healthy subjects was prepared by differential ultracentrifugation. 125I-LDL was prepared by the ICl method of MacFarlane. Prior to incubation with 125I-LDL, cells were preincubated in a medium containing 5% lipoprotein-deficient serum obtained from a myxoedematous patient. Uptake and degradation of LDL by normal cells and FH heterozygous cells increased about 20% with 1.0 microgram/ml of L-triiodo-thyronine (T3) or L-thyroxine (T4). In FH homozygous cells, no increase of LDL receptor activity was observed with T3 or T4. These results suggest that thyroid hormones regulate LDL catabolism through the LDL pathway. PMID- 6276237 TI - [Effects of synthetic human parathyroid hormone (1-34) on plasma and urinary adenosine 3',5'-monophosphate and urinary phosphate and calcium in patients with hypoparathyroidism and pseudohypoparathyroidism (author's transl)]. PMID- 6276238 TI - [Anterior pituitary function before and after treatment in twenty-three patients with isolated adrenocorticotropic hormone (ACTH) deficiency reported in Japan (author's transl)]. AB - Anterior pituitary function in 23 patients (16 men and 7 women, aged 27 to 68) with isolated ACTH deficiency was analyzed. Four were our own cases while the other 19 cases were ascertained by questionnaire. Both the baseline TSH levels and the peak TSH responses to TRH were high before treatment in more than half the cases but were normalized after treatment. This abnormality was found in patients younger than 50. The peak prolactin responses to TRH were excessive before and after treatment in three-fourths of the cases but decreased after treatment. The peak HGH responses to ITT were excessive in 3 patients before treatment an increased after treatment in 5 out of 6 cases. The peak LH and FSH responses to LH-RH were low or high in 20-30% of cases, but these abnormal responses were reduced to half after treatment. These results demonstrate that many disorders of the anterior pituitary function were found in patients with isolated ACTH deficiency but that these disorders became normal after treatment. PMID- 6276239 TI - [Subclinical hypothyroidism associated with the positive perchlorate discharge test in aged subjects (author's transl)]. PMID- 6276240 TI - [Studies on insulin metabolism: enzymatic characteristics and biological significance of insulin-degrading activities in rat liver cell fraction (author's transl)]. PMID- 6276241 TI - Cardiac emergencies. AB - Learning to effectively treat cardiac emergencies is not an easy task. It is my opinion that mastery of material presented herein will ensure minimal competency. Finding practical experiences to help develop the necessary medical skills will require imagination and creativity. The following are offered as possible resources:1. Attend the American Heart Association's basis and advanced life support classes. (CPR training must be repeated at six-month intervals.) 2. Spend several days and/or nights in the emergency room at a local hospital observing (helping with(emergency care. #. Spend time with a mobile coronary care unit if available in your community. 4. Watch and do venipunctures at the local blood bank or donor center. 5. Observe in the intensive care unit at a local hospital. 6. Assist an oral surgeon in preparing intravenous lines and monitor a patient during sedation. 7. Observe administration of oxygen by an anesthesiologist in the operating room at a local hospital. PMID- 6276242 TI - Spin-labelling of DNA with hydrazine mustard spin label (HMSL). AB - 1. The hydrazine mustard spin label (HMSL), recently synthesized in our laboratory (Raikova, 1977) was used for spin-labelling of DNA. 2. It alkylates both double- and single-stranded DNAs. 3. The reaction of HMSL with DNA was studied with respect to the kinetics of alkylation, dependence on salt concentration and base specificity. 4. It was found that HMSL is a base-specific reagent, alkylating preferentially guanine. According to their ability to bind HMSL, the four deoxyribonucleotides are ordered in the following way: G greater than A greater than C greater than T. 5. The EPR spectra obtained strongly depended on the secondary structure of the spin-labelled DNA: unlike the immobilized spectra of the double-stranded DNAs (2AZZ = 44.8G), the EPR spectra of single-stranded DNAs were non-immobilized (2AZZ = 32.8 G). 6. When sheared double-stranded DNA was spin-labelled, the parameters of the EPR spectrum depended also on the GC content of DNA. PMID- 6276243 TI - Isolation of nucleoside phosphotransferases from chromatin of Morris hepatoma 9121 nuclei. PMID- 6276244 TI - Heterogeneity of juvenile hormone binding compounds in fat bodies of a cockroach. AB - Fat bodies of adult vitellogenic or non-vitellogenic females and those of adult males contain 2 populations of saturable JH binding compounds. The respective Kd vales are 2-5 x 10(-9) M and 1-4 x 10(-8) M. Fat bodies of penultimate nymphs, hemolymph of any animal, and midgut epithelia contain one saturable JH binder with a Kd of 1-5 x 10(-8) M. Unlabeled JH I and III, as well as the JH analog ZR 515 compete effectively for the binding sites in these tissues. The 2 classes of hormone binders were separated by DEAE ion-exchange column chromatography. The very high affinity molecules of the adult fat bodies are presumably the specific JH receptors which may function in translocation of the hormone to the nucleus. The compound with relatively lower hormone affinity may be the JH hemolymph carrier which is also produced by the fat bodies and then exported. PMID- 6276245 TI - Properties of LH/hCG receptors in porcine corpus luteum homogenates and subcellular fractions, and factors influencing the recovery of membrane-bound hormone. AB - An evaluation of several techniques commonly used to separate bound from free hormone was made for the binding of radiolabelled human chorionic gonadotropin to mid-luteal pig corpus luteum homogenates and subcellular fractions. Compared to millipore filtration with 0.22 or 0.45-mu HAWP filters, centrifugation of homogenates at 2 500 x gav or 1 000 x gav for 15 min recovered only 50 and 33% respectively of the total bound hormone present. This discrepancy was accentuated further when hormone binding to post-nuclear subcellular fractions was studied. Centrifugation of a nuclear fraction at 2 500 x gav for 15 min recovered 70% of the membrane-bound receptors present (as measured by filtration), whereas centrifugation of mitochondrial, lysosomal and microsomal fractions under these conditions recovered only 46, 16 and 4% respectively. Divalent metal ions influenced hormone binding in 2 ways: (i) Specific bonding of hCG was greatest in the absence of metal ions and in the presence of low levels of chelating agents. Increasing concentrations of magnesium and calcium ions appeared to interfere with the interaction of hormone and receptor. (ii) Divalent metal ions reduced the recovery of particle-bound hormone by centrifugation, at both high and low speeds. The results illustrate the pitfalls of using low-speed centrifugation to recover membrane-bound hormone-receptor complexes when small membrane vesicles are present. PMID- 6276247 TI - hCG-induced inhibition of testicular steroidogenesis: an oestradiol-mediated process? AB - Single i.v. injections of hCG (10 U) in adult male rats resulted, within 24 h, in a 2-fold decrease in the maximal LH-stimulated testosterone production in vitro, while pregnenolone production was not changed. In addition to the changes in steroidogenesis, a concomitant depletion of the oestradiol-cytosol receptor was observed. In contrast with the observations after 24 h, a 2-fold increase in both testosterone and pregnenolone production was observed at 72 h after a single i.v. injection of hCG (10 U). At 72 h after the hCG treatment, oestradiol-cytosol receptor levels were not different from values observed after injection of saline. Tamoxifen administration (50-500 microgram s.c. injections) resulted, within 24 h, in depletion of oestradiol-cytosol receptor levels. This decrease of oestradiol binding, however, was not paralleled by decreases in testosterone production. Simultaneous administration of tamoxifen and hCG did not prevent the hCG-induced inhibition of testosterone production observed 24 h after administration of hCG. It is concluded that the present results offer no proof for an obligatory role of the oestradiol receptor in gonadotropin-induced inhibition of testosterone production in testicular Leydig cells. PMID- 6276246 TI - Hormonal control of phosphodiesterase activity in cultured rat Sertoli cells. AB - Phosphodiesterase activity was studied in cultures derived from 19-day-old rats and enriched with Sertoli cells. Pretreatment of such cultures with follicle stimulating hormone or L-isoproterenol increased cAMP-phosphodiesterase activity 5.2 and 2.0 times, respectively. cGMP-phosphodiesterase activity was not affected. Similar effects were observed in freshly isolated cells. The stimulatory effect was enhanced by the phosphodiesterase inhibitor 3-isobutyl-1 methylxanthine and was mimicked by cholera toxin and dbcAMP. Increased activity was observed after a latent period of 1 h. Stimulation was blocked by cycloheximide and actinomycin D. The enzyme had an apparent Km for cAMP of 1.4 micro M. Its activity was enhanced by Mg2+ but not by Ca2+. It is concluded that phosphodiesterases play an important role in the hormonal control of Sertoli-cell function and may contribute to the refractory state of these cells after stimulation with various agonists. PMID- 6276248 TI - Characterization of cytoplasmic ecdysteroid receptors in the hypodermis of the crayfish, Orconectes limosus. AB - [3H]Ecdysone, [3H]20-OH-ecdysone and [3H]ponasterone A were specifically bound by the 120 000 g supernatant of the homogenate from crayfish hypodermis. According to the HPLC test, the bound hormone was not metabolized. Analysis of the cytosol receptor by sucrose-density-gradient centrifugation, gel filtration and enzymatic degradation revealed the protein character of the receptor, a molecular weight of 70 000-80 000 D, and 5S. The binding capacity was rapidly lost after treatment with N-ethyl-maleimide or heating for 10 min at 60 degrees C. Optimal conditions for the demonstration of the receptor were incubation at 0-8 degrees C for 30 min and a salt concentration of 120 mM. A sequence of moulting hormones, and steroid hormones without moulting hormone activity were tested for ligand specificities. Estimation KD values yielded 3-6 x 10(-11) M for ponasterone A, 2-3 x 10(-9) M for 20-OH-ecdysone and 2 x 10(-8) M for ecdysone. PMID- 6276249 TI - Prolactin receptor turnover in explants of pseudopregnant rabbit mammary gland. AB - Pseudopregnant rabbit mammary glands in organ culture were used to investigate prolactin (PRL) receptor turnover. Chloroquine (100 microM) results in an increase in prolactin receptor levels (15.7 +/- 1.2% to 35.9 +/- 3.5% specific binding), whereas cycloheximide (1 microgram/ml) induces a rapid decline (to 6.4 +/- 1.2%) suggesting a rapid synthesis and degradation of the receptor molecule. Inhibitors of cellular transcription have little effect on receptor levels. Neither actinomycin D nor dichlororibofuranosylbenzimidazole (DRB) diminish PRL receptor levels whereas total protein synthesis is almost completely inhibited, and chloroquine increases the binding even in the presence of transcriptional inhibitors. These results imply that receptor synthesis continues and that the mRNA for the receptor protein is particularly stable. Ouabain (3 micrometers), which blocks the ATP-dependent Na+/K+ pump, provokes a greater than 60% reduction in PRL receptor levels without modifying total protein synthesis. Dinitrophenol (DNP, 1 mM), an oxidative uncoupler, has little effect on receptor levels, possibly due to a blockage of both synthesis and degradation. Prolactin is capable of inducing a 60% down-regulation of its own receptor, and this phenomenon appears to be energy-dependent because it is partially inhibited by DNP. This process seems to involve an increased rate of receptor degradation. These studies suggest that, at any one time, the level of PRL receptors in a target cell is the result of a dynamic equilibrium between receptor synthesis and degradation and that the most frequent modulations occur at the level of translation and lysosomal degradation. In conclusion, in mammary glands of the pseudopregnant rabbit, the prolactin receptor molecule appears to have a short half-life; the mRNA for this protein, however, is relatively stable. PMID- 6276250 TI - Specificity of the cyclic GMP-binding activity and of a cyclic GMP-dependent cyclic GMP phosphodiesterase in Dictyostelium discoideum. AB - The nucleotide specificity of the cyclic GMP-binding activity in a homogenate of Dictyostelium discoideum was determined by competition of cyclic GMP derivatives with [8-3H]-cyclic GMP for the binding sites. The results indicate that cyclic GMP is bound to the binding proteins by hydrogen bonds at N2H2, N1-H and/or C6 = O, N7, 2(1)-OH and 3(1)-O and possibly via a charge-charge interaction with the phosphate moiety of cyclic GMP. Cyclic AMP only competes with cyclic GMP for binding at a 20,000-fold higher concentration. The same cyclic GMP derivatives were used to modify the hydrolysis of [8-3H]cyclic GMP by phosphodiesterase. The phosphodiesterase is activated by cyclic GMP. The nucleotide specificity of activation of the enzyme differs from the specificity of the enzyme for hydrolysis. This indicates that activation by cyclic GMP and hydrolysis of cyclic GMP occur at different sites of the enzyme. Cyclic AMP neither activates the cyclic GMP phosphodiesterase nor competes with cyclic GMP for hydrolysis. This indicates that intracellular cyclic AMP does not interfere with the action of intracellular cyclic GMP in D. discoideum. PMID- 6276251 TI - Characterization of angiotensin II receptors in the anterior pituitary gland. AB - Specific and high affinity binding sites for angiotensin II were demonstrated in the anterior pituitary gland by binding studies with [125I] iodoangiotensin II. The binding properties of the pituitary receptors were similar to those of angiotensin II receptors present in the adrenal gland. The concentration of binding sites in rat anterior pituitary (293 +/- 50 fmoles/mg protein) was less than in the adrenal gland, but was much greater than in smooth muscle. Angiotensin II receptors were identified in the anterior pituitary tissue of mature and immature animals of both sexes, and in species including rat, rabbit and dog. No binding of angiotensin II was detected in posterior pituitary homogenates, or in GH3 pituitary tumor cells. Collagenase-dispersed anterior pituitary cells also contained specific binding sites for angiotensin II, with equilibrium binding constant (Ka) of 3.6 x 10(9) M-1. The presence of specific high-affinity angiotensin II receptor in the anterior pituitary gland provides a mechanism by which angiotensin-like peptides could modulate the process of pituitary hormone secretion. PMID- 6276252 TI - Impaired metabolic response to nerve stimulation in brown adipose tissue of hypothyroid rats. AB - In brown adipose tissue of the rat, chemically or surgically induced hypothyroidism caused the following effects. A large decrease of the magnitude of the metabolic response to electrical nerve stimulation. The deactivation half time of the response was reduced to 70% of the control value, with no change in catechol O-methyltransferase activity. Pre-incubation of tissues with norepinephrine, 10(-5) M, increased the response to subsequent nerve stimulation almost to that of the controls. The catecholamine analogue dose-response curves were shifted to the right. The shift was very pronounced for isoproterenol (K50 426 nM versus 2 nM), somewhat less marked for norepinephrine (7373 nM versus 194) and very slight for phenylephrine (2803 nM versus 1649); there was almost no change in Emax values. An increase of octanoate oxidative capacity. A decrease of the capacity of the stereoselective binding of (-)-[3H]dihydroalprenolol of the high-affinity (Kd 2.0 nM) sites to a fourth of the control value and an increase by a factor of 2.9 of the Kd of the low-affinity binding sites. This decrease of binding to the beta-receptors was not sufficient quantitatively to explain the decrease in the metabolic response, suggesting the existence of an additional defective reaction which could occur between the binding to the beta-receptors and the activation of the triglyceride lipase. These results show that the sharp decrease of the metabolic response of brown adipose tissue to nerve stimulation has multiple causes. The findings are discussed in the context of the drastic decrease of cold resistance in hypothyroid rats. PMID- 6276253 TI - Presence and disappearance of nerve growth factor receptors on sensory neurons in culture. PMID- 6276254 TI - Sciatin: a myotrophic protein increases the number of acetylcholine receptors and receptor clusters in cultured skeletal muscle. PMID- 6276255 TI - Interactions between mesoderm and ectoderm in segment formation in the embryo of a glossiphoniid leech. PMID- 6276256 TI - Chemoresponsiveness to cAMP and folic acid during growth, development, and dedifferentiation in Dictyostelium discoideum. AB - Chemoresponsiveness to cAMP and to folic acid are monitored in growing, developing, and dedifferentiating amebae of the cellular slime mold Dictyostelium discoideum. Two semiquantitative assays are employed, one measuring the directed movement of cells up a gradient of chemoattractant ('chemotaxis' assay) and the other measuring the outward spreading of cells in response to a chemical stimulant distributed equally throughout the substratum ('spreading' assay). Vegetative amebae possess relatively insignificant levels of chemotactic responsiveness to cAMP. Six h after the initiation of development, at approximately the same time as the onset of aggregation, cells rapidly acquire chemotactic responsiveness to cAMP. During 'erasure', a dedifferentiation induced by resuspending aggregating cells in fresh nutrient medium, chemotactic responsiveness to cAMP is lost just after the erasure event. By the same chemotactic assay, it is demonstrated that vegetative amebae possess a significant level of chemotactic responsiveness to folic acid. Two h after the initiation of development, cells completely lose chemotactic responsiveness to folic acid. During erasure, cells reacquire chemotactic responsiveness to folic acid at approximately the same time that they lose responsiveness to cAMP. Dramatically different results are obtained by the spreading assay. When cells lose chemotactic responsiveness to folic acid early in development and when erasing cells lose chemotactic responsiveness to cAMP, they retain the spreading response to the two stimulants, respectively. The different results obtained for chemoreception employing the two assays are discussed in terms of molecular mechanisms, and a testable hypothesis is proposed for the possible roles of chemoresponsiveness and erasure in late morphogenesis. PMID- 6276257 TI - Adenosine 3' :5' monophosphate receptor sites in Xenopus laevis oocytes. AB - Adenosine 3':5' cyclic monophosphate (cyclic AMP) binds to proteins present in the 100,000 g supernatant fractions of full-grown oocytes of Xenopus laevis. Optimal pH for the binding is 4.0. Two receptor binding sites have been characterized by density gradient centrifugation as peaks with sedimentation constants of 4.6 S and 5.9 S. The apparent dissociation constants for the two cyclic AMP binding sites are 7 nM and 40 nM. The total cyclic AMP binding capacity of oocyte cytosol is 15.8 +/- 2.2 fmoles/oocyte and remains constant during meiotic maturation induced by progesterone. PMID- 6276258 TI - Cyclic AMP phosphodiesterase in Leydig cell preparations of the rat testis. AB - Enzymatic digestion of the interstitial tissue of early juvenile and adult rat testes resulted in an enrichment of the Leydig cell population. The cells of the intertubular preparation from adult testes were separated by centrifugal elutriation, according to differences in sedimentation velocity, a counter-flow centrifugation technique leading to 70% Leydig cell purity. Using this approach, it was possible to demonstrate that Leydig cells from adult testes contain only low affinity isoenzymes of cyclic AMP phosphodiesterase (PDE; E.C.: 3.1.4.17), an intracellular regulator of cAMP. Starch gel electrophoresis showed that the isozyme of cAMP PDe of Leydig cells is masked in crude testis homogenates due to the relatively low level of these cells in the total population. In Leydig cells, there are two different electrophoretic forms expressed which resemble two of eleven different molecular forms of cAMP PDE demonstrated for comparison in 21 different organs of the adult rat. An interstitial cell preparation from early juvenile testes, with a Leydig cell content of up to 20%, was also investigated electrophoretically with regard to molecular forms of cAMP PDE, the properties of which were characterized by kinetics analysis of cAMP hydrolysis. The results presented are discussed in relation to the onset of testosterone synthesis in Leydig cells of prepubertal rats leading to the initiation of male puberty. PMID- 6276259 TI - The effects of neuraminidase and gangliosides on ovarian LH/hCG receptors during rat development. AB - Ovaries of neonatal rats are not endowed with specific LH/hCG receptors up to 6-8 days of age. Treatment of ovarian membranes of the neonatal rat with neuraminidase results in a specific binding of radioactively labeled hCG, while an increase of hormone binding is observed after neuraminidase treatment of ovarian membranes of the 21-day-old rat. These changes in hormone receptor sites in the ovary are dependent on the neuraminidase concentration used and are due to a receptor with a dissociation constant (KD) of about 10(-9) M. The KD of the receptor in the LH/hCG sensitive ovary without neuraminidase treatment is about 10(-10) M. These results indicate the presence of two different LH/hCG receptors in the ovarian membrane. The unmasking effect of neuraminidase onto LH/hCG receptors indicate that ganglioside-like structures are responsible for the masking of receptors in the neonatal, insensitive rat ovary and also in the 21 day-old sensitive ovary. Ganglioside preparations are able to inhibit the binding, and the fractionation of ovary gangliosides results in a fraction with a rather high inhibition potency of LH/hCG binding to the receptor. It is hypothesized that the masked receptor in the sensitive period represent a store of receptors for the reconstitution of the ovarian cells with active receptors after internalization of the hormone-receptor complex. Thus the masking of the receptors in the early postnatal rat ovary could be a prerequisite for the female differentiation of hypothalamic centers. The observed neuraminidase effect in vitro could reflect a physiologic situation. Neuraminidase was found in the ovary, and during early postnatal development the neuraminidase activity pattern coincides with that of the ovarian LH/hCG receptor changes. PMID- 6276260 TI - Antiserum to SV40 T antigens from a rabbit inoculated with autochthonous cells transformed in vitro by SV40. AB - A morphologically transformed cell line was established after infection with SV40 of the primary culture derived from a kidney removed surgically from a rabbit. The transformed cells were free of detectable infectious SV40 particles but produced them upon fusion with permissive monkey cells, and produced SV40 specific large and small T antigens. Repeated subcutaneous inoculations of the transformed cells into the animal from which the parental kidney cells had been derived failed to result in tumor formation but raised a substantial level of antibodies to the SV40-specific large and small T antigens as well as to the 58,000 dalton molecular weight cellular protein (middle T antigen). PMID- 6276261 TI - Aryl hydrocarbon hydroxylase and its components in human lung microsomes. AB - Microsomal fractions were prepared from specimens of human tissues obtained at surgical resections of 16 patients suffering from different types of pulmonary tumors. The contents of cytochrome P-450 and the activity of aryl hydrocarbon hydroxylase in human samples were much lower than the levels in rodent lung microsomes. The content of cytochrome b5 and the activities of NADPH-cytochrome P 450 reductase and of NADH-cytochrome b5 reductase, however, corresponded to those of rodent lung microsomes, with mean values of 0.12 nmol/mg protein, 0.039 and 3.58 mumol/min/mg protein, respectively. PMID- 6276262 TI - Rapid growth of carcinoma of the colon. PMID- 6276263 TI - Involvement of the duodenum by gastric carcinoma. PMID- 6276264 TI - Lung cancer: a persistent challenge. AB - The most frequently diagnosed type of lung cancer now is probably adenocarcinoma. This cell type is not clearly related to smoking, but may reflect environmental and occupational factors. Staging systems assess the extent of primary and metastatic disease, aid in determining appropriate treatment, and suggest prognosis. Clinical stage correlates with survival rate, but most staging systems underestimate the extent of metastases in lung cancer. PMID- 6276265 TI - [Cardiovascular system changes in workers in toluene diisocyanate manufacture]. PMID- 6276267 TI - Interaction of lipoproteins with macrophages. PMID- 6276266 TI - Lack of association of cytomegalovirus with adenocarcinoma of the colon. AB - Tumor specimens from patients with adenocarcinoma of the colon or rectum were examined for the presence of cytomegalovirus (CMV), and specimens of normal mucosa from the same patients were studied in parallel. Frozen sections of 14 specimens were made and the presence of CMV mRNA assayed by in situ hybridisation using 3H-labelled CMV-DNA as a probe. Nine of these sections were also tested for cytomegalovirus antigens by immunofluorescence. No viral nucleic acids or antigens were detected. In addition to these direct approaches, the specimens were disaggregated and 19 were successfully cultured in various media over several months without yielding virus on any occasion. Areas containing epithelial cells were found in some cultures, foci of bipolar cells in others, while, in several, fibroblastic cells predominated. To ensure that any virus containing cells were not lost by this method, the disaggregated tumour and normal intestinal cells were directly co-cultivated and also fused with human embryo lung cells, which are permissive for cytomegalovirus replication. The resulting cultures were examined over two to three months for the presence of cytomegalovirus, and in no instance was virus found, despite attempted induction by iododeoxyuridine. Two fusion cultures became contaminated with cytomegalovirus, strain AD-169, which was being handled in the laboratory at the same time. The strain was identified by the pattern of viral DNA fragments produced by restriction endonuclease cleavage. Thus the accidental passage of virus in the heterokaryons did not alter its DNA and would further indicate the absence of any cytomegalovirus genomes in the adenocarcinoma cells. PMID- 6276268 TI - Lysosomal granules in leukemic monocytes and their relation to maturation stages. PMID- 6276269 TI - Histiocytic medullary reticulosis re-visited. PMID- 6276270 TI - Immunohistochemical marking of malignant fibrous histiocytoma and malignant histiocytosis. PMID- 6276271 TI - Malignant histiocytic disorders of the skin. PMID- 6276272 TI - The role of macrophages as effector cells. PMID- 6276273 TI - Intracellular "labyrinths" in acute leukaemia. Do they indicate the maturational ability of leukaemic cells? AB - The occurrence and ultrastructure of the so-called "labyrinths" were investigated in the bone marrow cells from 13 patients with preleukaemia and acute leukaemia. Except for one patient, labyrinths were found in the myeloid cells from patients with preleukaemia and acute myelogenous leukaemia. Labyrinths could not be detected in lymphoblasts from patients suffering from acute lymphoblastic leukaemia or in plasma cells from a patient with plasma cell leukaemia. In patients with preleukaemia and smoldering leukaemia, labyrinths could be detected in all differentiation forms, from myeloblasts to mature polymorphonuclear granulocytes. Since it is improbable that labyrinths develop in consequence of exogenous effects, their presence in the mature granulocytes points to the in vivo maturational ability of leukaemic precursors in certain cases of acute myelogenous leukaemia and preleukaemia. PMID- 6276274 TI - Culture of rat Sertoli cells isolated with a modified procedure. Morphological identification of cell population and cell reactivity. AB - The effects of growing of Sertoli cells isolated from rat seminiferous epithelium by a modified procedure, and responses of these cells to dBcAMP or FSH stimulation was estimated using morphological methods. The modified isolation procedure included repeated mechanical rinsing of tubule pieces with a modified EDTA containing Hanks medium. Moreover, streptornase instead DNase was added to the trypsine containing medium and this resulted in a better dispersion of tubule components. The culture conditions remained unmodified. A high degree homogeneity of the cultured cell population and an evident reactivity of these cells to dBcAMP and FSH was achieved. Furthermore, an observation of giant cells, visible in monolayer among the typical Sertoli cells, is discussed in this paper. Contrary to small number of myoid cells, being survived in the culture, these giant cells did not show positive reaction to alkaline phosphatase. PMID- 6276275 TI - Problems of the classification of tumors of the nervous system and the significance of electron microscopy. PMID- 6276276 TI - [Modulation by endogenous opioid peptides of non-adrenergic neurotransmission in the guinea-pig taenia coli (author's transl)]. AB - The effects of opioid peptides on th response to stimulation of adrenergic or non adrenergic inhibitory nerve were investigated in the guinea-pig taenia coli. Leu enkephalin, met-enkephalin, (D-ala2) met-enkephalin and beta-endorphin inhibited non-adrenergic inhibitory response, in a dose-dependent manner. (D-ala2) metenkephalin and beta-endorphin were the most potent compounds in inhibiting the elicited relaxation, having ID50 values of 1 micro M. The inhibitory actions of these opioid peptides were reversed by naloxone (1 micro M). Naloxone itself potentiated the non-adrenergic inhibitory response. On the contrary, the adrenergic inhibitory response was not affected by application of leu-enkephalin, beta-endorphin or naloxone. Using an immunohistochemical method, met-enkephalin immunoreactive nerve fibers were observed in the longitudinal muscle layer, Auerbach plexus and circular muscle layer of the taenia. From these observations, it is concluded that endogenous opioid peptides may play a regulatory role in non adrenergic inhibitory neurotransmission but endogenous opioid peptides may play a regulatory role in non-adrenergic inhibitory neurotransmission but not in adrenergic neurotransmission. PMID- 6276277 TI - Effects of dietary bran and the colon carcinogen 1,2-dimethylhydrazine on faecal beta-glucuronidase activity in mice. PMID- 6276278 TI - [Treatment of chronic constipation with swelling substances]. AB - In 20 patients with chronic constipation and, in many cases, years of laxative abuse, in a prospective and randomized study, treatment with new, specially prepared swelling substances administered at the recommended doses, was carried out after first stopping the use of laxatives. After only one week of this treatment, a marked increase in the weight of the stools, and a reduction in stool transit time (p less than 0.05 in each case) were observed. PMID- 6276279 TI - [Pathomechanism of cholestasis]. PMID- 6276280 TI - [Clinicopathological studies of DNA content in nuclei of cells isolated from patients with esophageal carcinoma (author's transl)]. PMID- 6276281 TI - [A microangiographic study on the vascular pattern of the stomach in early carcinoma with particular reference to the color change in gastric endoscopy (author's transl)]. PMID- 6276282 TI - Experimental study on segmental demyelination in tellurium neuropathy. AB - Weanling rats fed a diet containing elemental tellurium became paralyzed of their hind legs based on segmental demyelination of the sciatic nerves. Recovery from the paralysis and remyelination took place despite continued receiving of the diet. The author could divide the process of demyelination and remyelination into four stages (Stage I-IV). The earliest changes were observed in the Schwann cell cytoplasm in which the Golgi complex revealed shrinkage, fragmentation and vacuolation. These changes soon involved the endoplasmic reticulum (Stage I). Degenerative changes of the Golgi complex and the endoplasmic reticulum were considered to disturb the active Schwann cell metabolism inhibiting the synthesis and maintenance of the myelin, resulted in disintegration and destruction of the myelin sheath (Stage II). Schwann cells proliferated around the demyelinated axons extending numerous elongated processes (Stage III). When remyelination began, only one simple shaped Schwann cell was associated with the axon. Remyelinated fibers were generally small in size, which was considered to be related to the formation of the short segments as well as to the Schwann cell proliferation (Stage IV). PMID- 6276283 TI - [Cyclic AMP accumulation in psoriatic skin: differential responses to epinephrine, AMP and histamine (author's transl)]. AB - Epidermal adenylate cyclase can be activated independently by epinephrine, adenosine and histamine resulting in the accumulation of cyclic AMP. Using the uninvolved and involved keratome-sliced skin from psoriatic patients, we investigated the effects of these agents in vitro on the intra-cellular cyclic AMP levels of the skin. In the involved skin of psoriasis, epinephrine-induced cyclic AMP accumulation was decreased, whereas no decrease in adenosine- or histamine-induced cyclic AMP accumulation was seen. Since keratome-sliced skin samples had various amounts of dermal contamination, we also investigated the effect of epinephrine on the "pure" epidermal cyclic AMP level. After the incubation with epinephrine, pure epidermal samples, which were micro-dissected free from stratum corneum, dermis and skin appendages, were assayed for cyclic AMP level. Again cyclic AMP accumulation was decreased in the involved skin. Thus epinephrine-induced cyclic AMP accumulation was shown to be decreased in the involved epidermis of psoriasis. PMID- 6276284 TI - Impaired hormonal regulation of adenosine 3',5'-monophosphate release in adipose tissue from hyperglycemic sand rats in vitro. AB - Egyptian sand rats (Psammomys obesus) developed under laboratory holding conditions and in dependence of different food intake various states of hyperglycemia and hyperinsulinism. Associated with the development of this syndrome insulin resistance of muscle- and adipose tissue appears. The question arose as to whether hyperglycemia and hyperinsulinism generally leads to hormone resistance. Therefore, we selected sand rats with different degrees of hyperglycemia and various stages of hyperinsulinism. Two groups of sand rats were investigated: normoglycemic and hyperglycemic animals. In these sand rats the catecholamine action on cAMP production adipose tissue was studied in vitro. There was a significant reduction of hormone responsiveness in adipose tissue of the hyperglycemic group compared with the noradrenaline response of adipose tissue of the normoglycemic sand rats. In no case had insulin significantly inhibited the noradrenaline stimulated cAMP production. By means of adenosine deaminase studies the interference of the eventual release of adenosine with the hormone action on adipose tissue in vitro could be excluded. In the sand rats correlations between IRI levels in the decapitation blood and noradrenaline stimulated cAMP release associated with simultaneous occurrence of insulin- and catecholamine resistance in adipose tissue seem to indicate that hormone resistance could be a general phenomenon during the development of hyperinsulinism. PMID- 6276285 TI - Alterations of plasma levels of pituitary hormones after oral application of metyrapone. PMID- 6276286 TI - Effects of a Met-enkephalin analogue and naloxone infusion on anterior pituitary hormone secretion in acromegaly. AB - The effect of a long-acting analogue of Met-enkephalin (Damme) and naloxone on anterior pituitary hormone secretion has been investigated in 14 acromegalic patients. Damme produced a progressive fall in circulating LH and cortisol and stimulated prolactin release in normoprolactinaemic patients. Naloxone infusion significantly stimulated gonadotrophin and cortisol secretion without modifying basal prolactin release both in normo-and hyperprolactinamic patients. GH levels remained unchanged during naloxone and Damme infusion. The data suggest that endogenous opiate receptors do not play a major role in modulating GH hypersecretion in acromegaly. PMID- 6276287 TI - The mode of genetic transmission of gouty family with increased phosphoribosylpyrophosphate synthetase activity. AB - The mode of genetic transmission of gout and increased activity of phosphoribosylphrophosphate synthetase (PRPPS) was studied in one family. Among 15 members of Family F, two male members had gout and had PRPPS activity of erythrocyte lysates three times higher than normal subjects. Five female members had activity 2.5 times higher than normal. The difference between the activities of male and female affected members was statistically significant (P less than 0.05). To examine the genetic trait of this abnormal PRPPS, the incorporation of 3H-adenine into erythrocytes or lymphocytes was studied using autoradiography. The number of grains which show the uptake of labeled adenine into cells revealed a normal distribution pattern in two normal persons and in two male patients, and a mixed pattern of the two cell populations in two female affected members. These results suggested mosaicism in female members and X-linked dominant transmission of this trait. Thermal inactivation of PRPPS of an affected female was intermediate between that from a normal subject and that from the affected males. This result showed the heterogeneity of the PRPPS from the hemolysate of an affected female. The genotype of PRPPS on the X-chromosome was assumed and the lod score between PRPPS and Xg was also estimated. From these findings and electrophoretical study, it was suggested that the abnormal enzyme was a mutant enzyme transmitted in an X-linked dominant trait, and that the mutation occurred on the structural gene of the PRPPS. PMID- 6276288 TI - Expression of GALT in two unrelated 9p- patients. Evidence for assignment of the GALT locus to the 9p21 band. AB - The quantitative expression of GALT and galactose utilization have been investigated in two patients with 9p deletion. Case 1 had a distal deletion of the band 9p22 leads to pter, while case 2 had an interstitial deletion of the region 9p133 leads to p23. In the former patient GALT activity and galactose utilization were found to be normal: in the latter decreased GALT activity and a significant decrease of galactose utilization were present. The above findings suggest that the GALT locus is in the 9p21 band. PMID- 6276289 TI - Primary culture of normal rat adrenocortical cells. I. Culture conditions for optimal growth and function. AB - Rat adrenocortical cells retained their differentiated characteristics over 2 wk in culture without a specific requirement for additives other than inorganic salts, amino acids, vitamins, and fetal bovine serum. The cells were maintained free from fibroblast overgrowth by substitution of D-value in place of L-valine in the medium. Corticotropin (ACTH) inhibited the growth of adrenocortical cells in this medium and the effect was reversible. The adrenocortical cells had a limited capacity for growth as reflected by total cell counts and [3H]thymidine uptake with cells from young animals demonstrated a greater potential for DNA synthesis than cells obtained from mature animals. A very sensitive assay for ACTH using a small number of cells in primary culture also is described. PMID- 6276290 TI - Primary culture of supporting cells from bovine testes. AB - Supporting cells from 6- to 8-wk-old bovine calf testes were maintained in cell culture and were characterized morphologically and biochemically. Light and electron microscopy showed that the cultures consisted principally of supporting cells that were identified as epithelial cells with nucleolar vesicles. The bovine supporting cells in culture responded to follicle stimulating hormone with increased cAMP synthesis and enhanced incorporation of [3H]leucine into secreted proteins. No androgen binding protein could be detected in the medium. PMID- 6276291 TI - Regulation of uridine kinase activity in BALB/C-3T3 cells by serum components. AB - After the stimulation of quiescent density-inhibited BALB/c-3T3 cells with fresh bovine calf serum, uridine kinase activity measured in cellular extracts increased between hours 3 and 6 of incubation and remained elevated through 12 h after stimulation. The addition of either partially purified platelet-derived growth factor (PDGF) or platelet-poor plasma (PPP) also caused increased uridine kinase activity by 6 h, but the increased activity was not maintained and the activity returned to the prestimulated level by 12 h. However, when PDGF and PPP were added in combination an increased level of uridine kinase activity was maintained in a manner similar to that seen after the addition of serum. The components of PPP eluted in the void volume from Sephadex G-50 chromatography did not induce uridine kinase activity when present alone, although they did act synergistically with PDGF to allow the maintenance of elevated levels or uridine kinase activity over the period from 6 to 12 h after stimulation. Thymidine kinase activity was not induced by the addition of either PDGF or PPP alone, although either serum or the combination of PDGF and PPP did produce and induction of thymidine kinase activity in late G1. PMID- 6276292 TI - Hormonal effects of glucose-6-phosphatase in the liver & kidney of rats of different ages. PMID- 6276293 TI - A site-specific DNA methylase from Escherichia coli K. PMID- 6276294 TI - Mitogenic response of peripheral blood lymphocytes from patients with Graves' disease incubated with solubilized thyroid cell membranes containing TSH receptor and with thyroglobulin. AB - Transformation of peripheral blood lymphocytes (PBL) in response to solubilized TSH receptor and to human thyroglobulin (hTG) was carried out in patients with Graves' disease (GD). Mean stimulation index (SI) in response to solubilized TSH receptor was significantly increased only in hyperthyroid GD patients (SI = 3.0 +/- 1.8 SD, n = 13, p less than 0.025) when compared to normal subjects (SI = 1.5 +/- 0.8 SD,n = 14). All other subgroups of GD patients (i.e., euthyroid GD patients after treatment as well as those with/without ophthalmopathy or TSH displacing antibody [TDA]) showed a tendency to elevated mean SI, which, however, was never significantly increased. In contrast, patients who were suspected of having a "disseminated autonomy" showed a tendency to decreased mean SI compared to GD patients. The mitogenic response of PBLs induced by solubilized TSH receptor could be suppressed by incubation of antigen and PBLs with bTSH. Control experiments with purified protein derivate of tuberculin(PPD) as stimulating antigen revealed, however, that bTSH itself suppresses the mitogenic response of PBLs and that the decrease of SI in this experiment is not due to blockage of the antigenic determinants of the TSH receptor protein. Human thyroglobulin (hTG) produced no significant increase in the SI of PBLs. Only 9 out of 47 GD patients showed an SI higher than the mean of normals + 2 SD. All those patients, however, who showed an elevated SI ( greater than 2.0) also revealed an increased SI with solubilized TSH receptor. PMID- 6276295 TI - Cyclosporin A abrogates proliferation of T cells and generation of suppressor and cytotoxic T-cell function induced by Epstein-Barr virus. AB - Cyclosporin A (CYA) promotes the outgrowth in vitro of Epstein-Barr-virus(EBV) infected cells of immune donors. In the present study, the effects of CYA on the T-cell responses developed to an in-vitro EBV infection were studied. Cyclosporin A, by acting on the responder cells and not on stimulator cells, strongly inhibited the proliferation of T cells normally induced by EBV-infected autologous cells. Moreover, T cells from cultures not exposed to CYA exerted suppression on both alloantigen-induced DNA synthesis and PWM-stimulated immunoglobulin producton of autologous peripheral blood mononuclear cells. In contrast, T cells from cultures treated with CYA exhibited significantly less or no suppressor activity as determined in both indicator system. Finally, CYA abrogated the generation of cytotoxic T cells against EBV-infected autologous cells, whereas non-CYA -treated T cells killed the virus-transformed target cells. Both suppressor and cytotoxic T-cell functions are known to play an essential role in the control of EBV infection by limiting the continuous growth of the virus-infected cells. These results, therefore, stongly suggest that cyclosporin A promotes the outgrowth of EBV-infected cells by abrogating the T cell responses to the Epstein-Barr virus. PMID- 6276296 TI - Induction of suppressor cells after activation of human peripheral blood mononuclear cells with sodium periodate. AB - Pretreatment of normal human peripheral blood mononuclear cells (MN) with sodium periodate (NaIO4) resulted in the induction of suppressor cells. The mitogenic response of fresh allogeneic and autologous cells to phytohaemagglutinin (PHA), Concanavalin A (Con A), pokeweed mitogen (PWM), various antigens and in mixed lymphocyte culture was suppressed when NaIO4 pretreated cells were present. PWM induced plaque-forming-cell responses were also suppressed by NaIO4-pretreated cells. Treatment of cells with mitomycin C before the NaIO4 treatment abolished the suppressive activity. A ratio of 1 : combination that resulted in the strongest suppression. Supernatants from NaIO4-pretreated cell cultures were not inhibitory. PMID- 6276297 TI - Management of Wilms' tumor and neuroblastoma. PMID- 6276298 TI - Oxidation of amino acids by human neutrophils. AB - We studied the oxidation of alanine and methionine by human neutrophils. Phagocytosis enhanced the decarboxylation of amino acids by human neutrophils. Decarboxylation of amino acids was dependent on the myeloperoxidase system (MPO- H2O2--Cl-). This was further confirmed using purified canine MPO. Human neutrophils and the MPO system were about 10 times more efficient in decarboxylating alanine than methionine. They also oxidized methionine to methionine sulfoxide. The fraction of methionine decarboxylated by human neutrophils or the MPO system was small compared to the fraction which was oxidized to methionine sulfoxide. Thus methionine was preferentially oxidized to methionine sulfoxide by the MPO system. However, once methionine was oxidized to methionine sulfoxide, it was readily decarboxylated by the MPO system. The results suggest that the thio group of methionine prevents its carboxylic group from being decarboxylated. PMID- 6276299 TI - Hepatoblastoma. PMID- 6276300 TI - Potentiating effect of bile on enterotoxin-induced diarrhea. AB - The influence of bile acids on adenosine 3',5'-phosphate-induced intestinal secretion was studied in mice. Bile flow was stopped by ligation of the common bile duct, and secretion was induced in ligated loops of the small intestine. The decrease of bile led to inhibition of hypersecretion after challenge with heat labile enterotoxins from Vibrio cholerae and Escherichia coli, as well as with prostaglandin E1. In contrast, the fluid response induced by dibutyryl-adenosine 3',5'-phosphate was unaffected by intestinal bile. Injection of bile or bile acids into intestinal loops before cholera toxin challenge restored the toxin induced secretion in the bile-depleted intestine. At the subcellular level the decrease of intestinal bile led to inhibition of cholera toxin-activated adenylate cyclase, whereas the bile concentration did not influence the binding of 125I-labeled toxin to the intestinal epithelial cells. The results suggest that intestinal bile interacts with adenylate cyclase in the induction of fluid secretion by enterotoxins and prostaglandin E1. PMID- 6276301 TI - Cross-protection in nonhuman primates against Argentine hemorrhagic fever. AB - The susceptibility of the marmoset Callithrix jacchus to Tacaribe virus infection was investigated to perform cross-protection studies between Junin and Tacaribe viruses. Five marmosets inoculated with Tacaribe virus failed to show any signs of disease, any alterations in erythrocyte, leukocyte, reticulocyte, and platelet counts or any changes in hematocrit or hemoglobin values. No Tacaribe virus could be recovered from blood at any time postinfection. Anti-Tacaribe neutralizing antibodies appeared 3 weeks postinfection. The five Tacaribe-infected marmosets and four noninfected controls were challenged with the pathogenic strain of Junin virus on day 60 post-Tacaribe infection. The former group showed no signs of disease, no viremia, and no challenge virus replication, whereas the control group exhibited the typical symptoms of Argentine hemorrhagic fever, high viremia, and viral titers in organs. Soon after challenge, the Tacaribe-protected marmosets synthesized neutralizing antibodies against Junin virus. These results indicate that the marmoset C. jacchus can be considered an experimental model for protection studies with arenaviruses and that the Tacaribe virus could be considered as a potential vaccine against Junin virus. PMID- 6276302 TI - Experimental parainfluenzavirus infection in mice: fatal illness with atrophy of thymus and spleen in mice caused by a variant of parainfluenza 3 virus. AB - Intracerebral inoculation of the YN strain of parainfluenza 3 virus was found to induce an acute fatal illness characterized by marked thymic and splenic atrophy in newborn mice. Previously we showed that the YN strain contains three distinct plaque-type variants, LT, SC, and M. Of these, the M-type variant induced this fatal illness, whereas the other two variants induced hydrocephalus. PMID- 6276303 TI - Alteration of granuloma angiotensin I-converting enzyme activity by regulatory T lymphocytes in murine schistosomiasis. AB - Granulomatous inflammatory lesions in murine schistosomiasis mansoni undergo a spontaneous diminution at the chronic phase of the disease concurrent with an increase in angiotensin I-converting enzyme (ACE) activity. The objective of this investigation was to determine whether T cells influence ACE activity within the granulomas. Cyclophosphamide and cimetidine treatments of mice, which augment delayed-type hypersensitivity reactions, enhanced liver granuloma size and decreased granuloma ACE activity. The suppression of liver granuloma by the adoptive transfer of spleen cells from chronically infected, immunomodulated mice into recipients with acute infection increased ACE activity of granulomas and granuloma macrophages. The increased ACE activity was dependent upon the transfer of Thy 1.2+ splenic lymphocytes. The level of the ACE activity was proportionate to the number of administered spleen cells. Thus, the level of ACE activity within the egg-induced granuloma is dependent upon T lymphocytes present in the spleen and granulomas of infected animals. PMID- 6276304 TI - Physiology of "mutans-like" Streptococcus ferus from wild rats. AB - Strains of Streptococcus ferus isolated from the oral cavities of wild rodents inhabiting sucrose-rich and sucrose-poor environments have many traits in common with the "mutans" streptococci. Thus, S. ferus HD3 and 8S1, like cariogenic S. sobrinus 6715-13, from adherent, alpha (1 leads to 3) glucopyranosyl-glucose linkage-rich, plaquelike deposits in vitro and in vivo through the action of constitutive glucosyltransferase(s) enzymes on sucrose, produce and degrade intracellular polysaccharide, produce short-chain fatty acids from the catabolism of mono- and disaccharides, carry the c antigen of S. mutans, and penetrate, persist, and proliferate in a sucrose-augmented fashion in the oral cavities of specific-pathogen-free rodent caries models. However, unlike infection with common S. mutans, infection with tested S. ferus strains does not cause caries. This avirulence appeared to result more from the reduced aciduricity of S. ferus than from differences in glucosyltransferase complements. Studies showed that despite generally similar growth rates and extracellular glucan syntheses, the acidogenic metabolism of S. ferus was more inhibited by declining environmental pH than was cariogenic S. sobrinus 6715-13 and that, in vitro, less hydroxyapatite was solubilized by S. ferus metabolic end products. The physiology of these S. ferus strains demonstrated that, in addition to plaque formation and acid production, acid tolerance was crucial to the carious process. PMID- 6276305 TI - Delayed hypersensitivity to herpes simplex virus: murine model. AB - Cell-mediated immunity has been shown to be clinically important in recovery from herpes simplex virus (HSV) infections. To investigate the role of delayed hypersensitivity (DH) in immunity and protection against HSV, we developed a murine model using the ear-swelling assay. Mice were infected subcutaneously with HSV-1 and ear-challenged, and the swelling was quantified. Significant ear swelling was detected by 3 to 4 days postinfection and peaked at 6 days. The kinetics of development of ear swelling were typical of DH: maximal swelling occurred 24 h post challenge and was diminished by 48 h, and the cellular infiltrate was predominantly mononuclear. Four-hour swelling, indicative of antibody-mediated, immediate-type hypersensitivity, was not detected until 15 days post immunization. The DH response was virus specific and could be transferred to normal recipients with lymph node T cells, but not with B cells or immune serum. This system will provide a useful model for evaluating the protective role of DH in HSV infection and for studying the specificity and interaction of T cells which mediate the response. PMID- 6276306 TI - Polymyxin B suppresses the endotoxin inhibition of concanavalin a-mediated erythrocyte agglutination. AB - The lectin agglutinability of human erythrocytes has been utilized to examine interactions of gram-negative endotoxin with mammalian cell plasma membranes. Erythrocytes treated in buffer with Escherichia coli 0127:B8 lipopolysaccharide (LPS) or Salmonella minnesota Re595 glycolipid for 1 h became resistant to agglutination by the lectin concanavalin A (ConA) in buffer free of LPS or glycolipid. Polymyxin B, a cationic cyclic lipopeptide which specifically binds to the lipid A toxophore, was tested for possible effects on the LPS and glycolipid inhibition of Con A erythrocyte agglutination. The presence of polymyxin B during the initial 1-h treatment with LPS or glycolipid blocked the ability of the endotoxins to render erythrocytes refractory to agglutination by ConA. Inhibition by polymyxin B was stoichiometric, and in repeated experiments, LPS was completely suppressed in the hemagglutination assay at a polymyxin B:LPS weight ratio of 1:4.1 (increasing polymyxin concentration, constant LPS concentration) and 1:5.1 (constant polymyxin concentration, increasing LPS concentration). These stoichiometry values are similar to values obtained for inhibition by polymyxin B of LPS lymphoid cell activation. It was concluded, therefore, that endotoxin inhibition of ConA erythrocyte agglutination reflects interactions of erythrocyte membranes with the lipid A region of endotoxin. In addition, the stoichiometry of polymyxin B inhibition suggests a similar extent of lipid A-dependent LPS interaction with erythrocytes and lymphoid cells. PMID- 6276307 TI - Boosterable IgE antibody response in mice without the use of adjuvant. AB - A mouse model is presented giving boosterable IgE antibody responses preceding the IgG antibody responses without using adjuvants. Groups of CBA mice were injected subcutaneously with minute doses of penicilloyl bovine gammaglobulin, 3.8-94 ng daily for one or two 10-day periods. Some groups of animals were simultaneously given Bordetella pertussis bacteria intraperitoneally. The IgE antibody responses were recorded with passive cutaneous anaphylaxis (PCA) in rats as well with RAST. IgG antibody responses were determined with a double antibody assay. An overall good agreement between the antibody activity in the PCA and RAST was seen, both tests showing boosterable IgE antibody responses. Some discrepancies between the two tests are discussed. PMID- 6276308 TI - Is eosinophil chemotactic factor identical with leukotriene B? AB - Eosinophil chemotactic factor (ECF) is a chemotactic factor for eosinophils and neutrophils that is non-peptide in nature. By several lines of evidence, this substance is closely similar or identical to 5,12-di-HETE (leukotriene B). Common denominators induce their derivation from arachidonic acid, their cells of origin, thier physiochemical characteristics and their biological activity. Differences in the ECF activity generated by different types of leukocytes suggest, however, that isomers of leukotriene B or as yet unidentified molecules might be involved. For this reason, ECF should not be called leukotriene B at this point of time. PMID- 6276309 TI - Ketoconazole (R 41 400) in the treatment of dermatophyte infections: a clinical, mycological, and morphological study. AB - Ketoconazole (100 mg, orally, once daily) was investigated in nine patients with extensive dermatophyte infections. After treatment ranging between one week and three months, clinical cures (healing of lesions and negative cultures) were observed in all cases. In vitro growth of Trichophyton rubrum, T. mentagrophytes, and T. verrucosum isolated from infected skin scales were completely inhibited by concentrations of ketoconazole of 10 microgram/ml and above. No evidence for the development of drug resistance was obtained from regular in vitro sensitivity tests. PMID- 6276310 TI - Assays for transferrin and transferrin receptors in tumour and other mouse tissues. PMID- 6276311 TI - The role of transferrins in gallium uptake. PMID- 6276312 TI - Relationships between the metabolism of 67 Ga and iron. PMID- 6276313 TI - Studies on the uptake of 67 Ga and 59 Fe and the binding of transferrin by cultured mouse tumour cells. PMID- 6276314 TI - Vitamin D activity of different vitamin D3 esters in chicken, Japanese quail and in rats. PMID- 6276315 TI - Calcinogenic activity of vitamin D3 and vitamin D3 palmitate in rat and rabbit. AB - Experimental hypervitaminosis D was produced in rabbits by feeding 25,000 I.U. vitamin D3 or the corresponding amount of vitamin D3 palmitate per kg of diet. Hypercalcemia and hyperphosphatemia was accompanied by increased CaBP activity and reduced weight gain in the vitamin D3 group as well as in the vitamin D3 ester group. The degree of calcification in the aorta and in the kidney also was similar in both groups. Increasing the vitamin intake by giving 10,000 I.U. vitamin D3 or vitamin D3 palmitate per day resulted in earlier and more widespread calcific deposits. In rats, receiving 50,000 I.U. vitamins D3 or vitamin D3 palmitate per kg of diet, calcification in soft tissue was much less extensive than in rabbits. But again, no difference was seen between vitamin D3 and vitamin D3 palmitate. These results indicate, that vitamin D3 esters are not suitable as a less calcinogenic form of vitamin D3. PMID- 6276316 TI - Serum calcium, phosphorus and magnesium responses to massive dosing of cholecalciferol (CC) and 25-OH-CC in young and aged ewes. AB - Twenty-four ewes were divided into two age groups (one-year-old and nine-year old) and used to determine the influence of cholecalciferol (CC) and 25-OH-CC on serum concentrations of Ca, P, and Mg with time post-injection. The ewes were maintained in slatted-floor pens and fed 800 g per head daily of a diet which analyzed 0.38% calcium, 0.31% phosphorus, and 0.14% magnesium. This diet was fed throughout the 21-day trial. The ewes were injected on days 0 and 7 as follows: (control) 5 ml ethanol; (CC) 50 mg CC in 5 ml ethanol; and (25-OH-CC) 25 mg 25-OH CC in ethanol. Blood samples were pre-injection (day 0) and on days 1, 2, 3, 5, 7, 9, 12, 17 and 21 of the trial. Serum Ca averaged 10.81, 11.06 and 11.25 mg/100 ml for the one-year-old ewes and 10.51, 11.06 and 11.54 mg/100 ml for the nine year-olds across sampling times in groups A to C, respectively. Serum P across sampling times averaged 6.58, 7.80 and 9.51 mg/100 ml in one-year-old ewes and were different (P less than .05) from each other. Serum P averaged 7.06, 8.56 and 8.59 mg/100 ml for the nine-year-old ewes. Serum Mg values were 2.51, 2.32 and 2.19 mg/100 ml for the one-year-old and 2.38, 2.14 and 2.00 mg/100 ml for the nine-year-old ewes across all sampling times for control, CC and 25-OH-CC groups, respectively. Serum Mg in one-year-old ewes was lower (P less than .05) in both CC and 25-OH-CC injected ewes than controls, and was lowest (P less than .05) for 25-OH-CC when compared with the control in nine-year-old ewes. External symptoms of hypervitaminosis (reduced feed intake and leg abnormalities) were apparent after the second injection with 25-OH-CC, and were most pronounced in the aged ewes. PMID- 6276317 TI - The new CDC Guidelines on Infection Control. PMID- 6276318 TI - CDC guidelines on infection control. PMID- 6276319 TI - Consumer education in discharge planning for continuity of care. AB - The need to establish continuity of care for patients in transition from the hospital; to community is essential for adequate community adjustment. The Resource Group model of discharge planning is based upon a consumer education model with emphasizes the responsibility of the individual in discharge planning. In these groups, the partial hospitalization program staff representative and the clinical pharmacist, in conjunction with inpatient clinical social workers, begin discharge planning with inpatients during the early phase of hospitalization. Increased utilization of partial hospitalization services has resulted in greater continuity of care. PMID- 6276320 TI - Studies on the source and release of collagenase in thermally burned corneas of vitamin A-deficient and control rats. AB - In previous studies we found that a mild thermal burn of the vitamin A-deficient rat cornea caused collagenase release into the medium of corneas placed in culture media 72 hr after applying the burn. Collagenase was released only on day 1 of culture and was not released from identically burned corneas of control rats. We now demonstrate that in deficient corneas, this collagenase release on day 1 of culture increased gradually with increasing time between burn and sacrifice, reaching a maximum at 16 hr after burning a remaining high up to 72 hr. In control rats day-1 collagenase release also increased to a maximum at 16 hr after the burn but then declined to almost zero at 72 hr. Trypsin treatment of day-1 media from both control and deficient corneas, taken at 72 hr after the burn, showed an almost complete absence of latent (inhibited) collagenase. Histologic observations revealed a close correlation between the presence of infiltrating polymorphonuclear neutrophils (PMNs) and the ulcerative lesions seen in burned, deficient corneas. When PMN infiltration was blocked by application of a tissue adhesive, no ulceration occurred and collagenase activity in the day-1 media dropped to almost zero. If burned and unburned areas of deficient corneas were separated and cultured separately, the burned area (containing most of the PMNs) was found to have 10 times the collagenase activity of the unburned area. In the controls, PMNs and some collagenase activity was detectable only 16 hr after the burn. We concluded that in the burned, vitamin A-deficient cornea there is increased attraction of PMNs to the lesion, resulting in collagenase release by these and possibly other cells, and ultimately resulting in ulceration. PMID- 6276321 TI - The effect of X-irradiation on Na-K ATPase and cation distribution in rabbit lens. AB - The Na-K ATPase activity of rabbit lens was measured at various times after exposure to a single dose of 2000 rads of X-ray and was compared with that in contralateral control eyes. A decrease in enzyme activity in both whole lens and in isolated capsule-epithelium was first observed 3 to 4 weeks after irradiation and became increasingly marked at 7.5 weeks after X-ray. These findings are consistent with our earlier observations that active transport of cations is reduced in these lenses and support the view that loss of membrane ATPase is responsible for the impairment of the cation pump in X-irradiated lenses. Despite a significant loss of the enzyme, X-irradiated lenses were able to maintain near normal levels of total cations (Na+ + K+), thus accounting for their normal hydration. The results of the changes in lens Na+ and K+ levels revealed that between 4 and 7.5 weeks after X-ray, the gain in Na+ was compensated by an equivalent loss of K+. A breakdown of this relationship of 1:1 exchange of Na+ for K+ is accompanied by a disproportionate increase in Na+ and water. PMID- 6276322 TI - Characteristics of BK papovavirus DNA prepared directly from human urine. AB - DNA was prepared directly from preparations pf papovavirus excreted in the urine of 2 patients on immunosuppressive therapy, without preliminary amplification in cell cultures. EcoRI, BamHI, and HindIII restriction patterns were typical of BK virus DNA and were identical for the two isolates. However, there were significant differences in the sizes of the HindIII fragments when compared with those of standard cell-culture-adapted BK virus strains. PMID- 6276323 TI - Identification of rotavirus particle types. AB - Negative-contrast electron microscopy of purified rotavirus particles reveals two particle types: single-shelled and double-shelled particles. The relationship of these particle types, seen by negative staining, to the enveloped and various types of nonenveloped particles seen in thin sections of virus-infected cells was determined. Thin-section and negative-contrast electron microscopic analyses were performed on cell lysates from simian rotavirus. SA11-infected cells and on highly purified double- and single-shelled particles. In thin sections, double shelled particles appeared as smooth-edged ovals containing dense nucleoids, whereas single-shelled particles had ragged edges and threads of material extending from their centers. The majority of nonenveloped particles seen in thin sections of infected cells were identified as double-shelled particles. Enveloped particles showed typical membrane structure and were observed rarely in crude rotavirus stocks, although they constitute about 10% of the particles within infected cells. It is hypothesized that the enveloped form is a transient one and the envelope is lost in the endoplasmic reticulum of the host cells. Finally, the 50-55 nm type IV particles seen within lysosome-like bodies in infected cells were identified as subviral particles formed from input virions. PMID- 6276324 TI - Inhibitory effect of retinoids on Epstein-Barr virus induction in Raji cells. AB - Induction of Epstein-Barr virus (EBV) early antigen after treatment with various combinations of croton oil and n-butyrate was markedly inhibited by retinoids 7901, 7902, Ro 10-9359 and Ro 11-1430. Possible administration of retinoids to virus capsid antigen IgA antibody-positive individuals in high-risk areas for nasopharyngeal carcinoma to prevent EBV activation and development of this cancer is discussed. PMID- 6276325 TI - Interaction of Herpes simplex virus with human cell lines at various stages of lymphoid differentiation. AB - The growth of herpes simplex virus (HSV) type 1 in human lymphoid cell lines, arrested at various stages of differentiation, was studied. The synthesis of viral DNA and antigens and the production of infectious virus were followed in null, B and T leukemia-lymphoma cell lines. It was found that while "nondifferentiated" null cells and cells differentiating along the B pathway, even at very early stages of differentiation, supported HSV growth, cell lines of T origin at all stages of differentiation were generally nonpermissive. The failure of T cells to support the growth of HSV did not result from inefficient adsorption to the host cells. However, viral DNA synthesis was not detected, and a significant reduction in the ability of synthesize HSV antigens was observed. It is suggested that the block in the growth cycle of the virus occurs at a stage after adsorption of the virion to the cells, but prior to synthesis of its DNA and proteins. PMID- 6276326 TI - Effect of protease inhibitors on the activation of Epstein-Barr virus repressed in cultured lymphoid cells. AB - The effects of five microbial and one synthetic protease inhibitors on the activation of Epstein-Barr virus expression in latency infected lymphoblastoid cells were studied. All protease inhibitors tested suppressed both viral antigen induction by 5-iododeoxyuridine treatment and cell growth in general. Among the inhibitors, however, antipain and pepstatin exerted a relatively strong and significant suppressive effect on antigen induction as compared to their inhibitory effect on cell growth. PMID- 6276327 TI - Epstein-Barr virus susceptibility of B cells correlates to the presence of complement receptors but not surface immunoglobulins. AB - Nuclear antigen (EBNA) inducibility by Epstein-Barr virus (EBV) was assessed in relation to the presence of complement (C3) receptors and surface immunoglobulins (SIg) on leukemic lymphocytes. When erythrocyte-antibody-complement complex(EAC) positive, SIg-negative B cells were exposed to EBV, EBNA synthesis occurred with a high frequency. In contrast, no significant EBNA induction was evident in SIg positive, EAC-negative B cells. Cells possessing both markers were EBV infectable, in proportion to the frequency of EAC-rosette formation. These data seem to indicate that EBV susceptibility of human B cells is closely related to the presence of C3 receptors but not SIg. Leukemic T cells and myelogenous leukemia cells were not EBV-susceptible, although some cells were significantly positive for C3 receptors. PMID- 6276328 TI - Replication of Heliothis zea baculovirus in an insect cell line. AB - The baculovirus, Heliothis zea nuclear polyhedrosis virus (HzSNPV), was grown in the H. zea cell line, IPLB-1075. Light microscopy observations showed that 95% of inoculated cells had CPE by 6 days postinoculation (p.i.). Growth curves of intracellular and extracellular virus showed that maximum titers (TCID50 units) were reached 3-4 days p.i. Studies on HzSNPV morphogenesis revealed that ultrastructural events were typical of baculovirus replication in vitro. EcoRI restriction enzyme analyses of viral DNA produced in larvae and in cell culture were identical and indicated that this virus was HzSNPV. PMID- 6276329 TI - Incorporation of 32P-phosphate into membrane phospholipids during infection of cultured human fibroblasts by herpes simplex virus type 1. AB - Infection of cultured human skin fibroblasts by herpes simplex virus leads to increased incorporation of labeled inorganic phosphate into host membrane sphingomyelin. Incorporation into the other major membrane phospholipids, including phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol and phosphatidylserine, was unaffected. Since sphingomyelin has been shown to serve as precursor to ceramides (via sphingomyelinase) in monkey kidney cells and since enhanced synthesis of ceramide-based glycolipids has been shown to occur after herpes simplex virus infection, we suggest that the observed increase in labeling of sphingomyelin may reflect mobilization for glycolipid synthesis. The distribution of phosphate label among the phospholipids of the viral envelope was identical to that among the phospholipids of the cellular cytoplasmic membrane fraction and differed from that of the nuclear fraction. PMID- 6276330 TI - Microscopic examination of the intracellular fate of infectious herpes simplex virus DNA. AB - The intracellular fate of 3H-labeled exogenous herpes simplex virus type 2 DNA following the exposure of recipient cells to selected DNA facilitators and the ultrastructural changes which ensued in transfected cultures were examined. In most cases, electron microscopic radioautography demonstrated that exogenous DNA reached the nucleus within 1 h postinoculation. At later time periods, enlargement of nuclei and margination of chromatin were noted. Cells treated with polyornithine differed significantly from those treated with other facilitators in that large cytoplasmic accumulations of labeled material associated with electron-dense, membrane-bound bodies persisted after 1 h postinoculation. In contrast, labeled DNA released by intact virions was detected in the nucleus as early as 20 min postinfection (p.i.), and progeny virus was evident by 8 h p.i. These results suggest that the efficiency of each facilitator is related to the rapidity with which exogenous DNA penetrates the recipient cell, traverses the cytoplasm, and the amount of DNA which is finally associated with the nuclear compartment. PMID- 6276331 TI - Resurgence of fetal isozymes in cancer: study of aldolase, pyruvate kinase, lactic dehydrogenase, and beta-hexosaminidase. PMID- 6276332 TI - Review: prostaglandins in diarrheal states. PMID- 6276333 TI - The effect of cyclic nucleotides on tolerance induction by dinitrophenyl isologous immunoglobulin G. AB - When tolerance to a hapten is induced by injecting the hapten conjugated to isologous immunoglobulin (Ig) G (dinitrophenyl (DNP)-IgG), the hapten remains on the surface of the antigen binding cell (ABC) for the duration of the tolerant state. Although this has been interpreted as a receptor blockade of ABC, it does not necessarily indicate that physical blocking of antigen receptors is the only mechanism of tolerance. Data are presented which suggest that cyclic nucleotides play a role in the in vitro induction of tolerance by DNP-IgG. Agents that elevate intracellular cyclic adenosine 3',5' monophosphate (cAMP) levels were added to lymphocytes during tolerance induction in vitro by DNP-IgG. Dibutyryl cAMP, theophylline, aminophylline, and isobutyl alpha-methylxanthine were able to inhibit the induction of tolerance to DNP. On the other hand, addition of dibutyryl cyclic guanosine 3',5'-monophosphate (cGMP) to lymphocytes incubated with doses of DNP-IgG that ordinarily are too low to induce tolerance facilitated tolerance induction. The possible role of these intracellular second messengers in the induction to tolerance is discussed. PMID- 6276334 TI - Restoration by cyclic guanosine monophosphate and extracellular calcium of butylated hydroxyanisole-suppressed primary murine thymus-dependent antibody response. AB - The effect of dibutyryl cyclic guanosine monophosphate (dbc-GMP) on butylated hydroxyanisole (BHA)-induced suppression of the primary in vitro thymus-dependent antibody response of BDF1 mouse spleen cultures was studied. When added at 0 hr relative to antigen addition, 1 mg of dbc-GMP (8 mM) restored by greater than 70% the BHA-inhibited primary immunoglobulin (Ig)M plaque-forming cell (PFC) response to sheep erythrocytes (SRBC). The suppression was not reversed by the addition of 50 microgram of dibutryl cyclic adenosine monophosphate (dbc-AMP), which is known to reverse suppressor T-cell activity. The addition of 10 mM extracellular calcium (Ca2+) at the same time as antigen to BHA-inhibited cultures resulted in more than 80% restoration of the anti-SRBC PFC response. Quantitation of c-GMP by radioimmunoassay demonstrated that BHA lowered by 58% the c-GMP content of splenic lymphocytes and abrogated the ability of lipopolysaccharide of E. coli (LPS) to elevate c-GMP levels in splenic lymphocytes. The data suggest that BHA exerts its immunosuppressive effect on the primary in vitro antibody response by inhibiting guanylate cyclase activity and effectively lowering c-GMP levels; exogenous dbc-GMP and Ca2+ can freely reverse the immunosuppressive effect of BHA. PMID- 6276335 TI - Acromegaly associated with neoplasia. (Description of a case and review of the literature). PMID- 6276336 TI - About recombinant deoxyribonucleic acid in medicine and industry. PMID- 6276338 TI - Skeletal muscle cytochrome c and myoglobin, endurance, and frequency of training. AB - This study was undertaken to evaluate the effects of various training frequencies on performance capacity, the mitochondrial marker cytochrome c, and myoglobin, which is responsible for storage and transport of O2, in the three types of skeletal muscle. Female rats were trained by treadmill running up to 120 min/day, either 2, 4, or 6 days/wk for 14 wk. As a result of training, exercise time to exhaustion was increased in proportion to the number of training sessions per week. Cytochrome c concentration increased (range 20-90%) as a linear function of the number of exercises per week in the fast-twitch red vastus lateralis (FTR), the slow-twitch soleus (STR), and the mixed plantaris muscles. However, the concentration of cytochrome c in fast-twitch white vastus lateralis (FTW) muscles increased to approximately the same extent (40-50%) in all training groups. The increases in myoglobin concentration (13-45%) with training were significantly related to frequency in FTR muscle but not in STR muscle. Myoglobin levels in FTW muscle remained unchanged, regardless of training group. These results provide evidence that the capacity to perform endurance exercise and the mitochondrial content of the red skeletal muscle types are directly affected by training frequency. PMID- 6276337 TI - Immunocytochemical detection of anti-ACTH reactivity in pancreatic islet cells of normal and steroid diabetic rats. AB - Morphologically different pancreatic islet cells of the rat reveal immunoreactivity under incubation with anti-ACTH 1-39. In normal rats, anti-ACTH reactivity is detectable predominantly in peripheral, polygonal cells. Paraffin serial section treated alternately with anti-ACTH and anti-glucagon show that the major portion of the anti-ACTH reactivity is localized in A cells. A few A cells, however, lack anti-ACTH reactivity. Anti-ACTH reactivity was also detected in individuals, round-to-oval, occasionally quite large and more centrally situated cells of pancreatic islets. In contrast, islets of steroid diabetic rats reveal a different distribution islets. In contrast, islets of steroid diabetic rats reveal a different distribution of anti-ACTH reactive cells. The number of peripheral reacting cells is greatly reduced; whereas there is an increase in the number of immunoreactive, large, round or polygonal cells, which are distributed throughout the islet. Preliminary investigations indicate that adsorption of ACTH 18-39 and glucagon onto anti-ACTH 1-39 reduces immunoreactivity in normal and steroid diabetic rats, whereas adsorption of ACTH 1-24 does not. PMID- 6276339 TI - Severe Epstein-Barr virus pulmonary involvement. AB - Two cases of severe Epstein-Barr virus (EBV) pulmonary involvement are described. Both were documented by EBV-specific serology and other causes of respiratory tract infection were excluded by culture and serology. Our first case describes fatal infectious mononucleosis (IM) with severe pneumonia in a patient without a definable immunologic disorder. Previous cases of pulmonary involvement in IM are reviewed. PMID- 6276340 TI - Health education in baccalaureate nursing curriculum: myth or reality? AB - This survey of baccalaureate nursing curricula was conducted to identify content and methods being used to help students develop beginning competence in health education. Catalogs from 200 schools revealed only minimal explicit attention to the teaching role of nurses. The 36 schools which gave evidence of health education and sent course materials showed little agreement on what health education encompassed or what reasonable expectations were for the baccalaureate nursing student. While there can be no generalization of these findings to baccalaureate nursing education, serious questions are raised concerning the thoroughness with which nursing educators are treating the health education component of the baccalaureate curriculum. Given the centrality of health education to prevention, health promotion, greater client control over his own health, reduced anxiety and more appropriate utilization, it would appear that health education is scarcely an area to be dealt with superficially in baccalaureate nursing education. PMID- 6276341 TI - Teaching through groups. PMID- 6276342 TI - Evaluating a curriculum. PMID- 6276343 TI - Advocacy-a tool for radical nursing curriculum planners. PMID- 6276344 TI - Contract grading in clinical evaluation. AB - The use of contracts in grading clinical performance has provided faculty with a solution to two important problems. One has been how to ensure that students are graded consistently among the various faculty who are grading the same clinical component of a course. The second has been to increase the opportunity for individualized attention to student learning needs. We have been pleased to see all the faculty members increase their input and involvement in the continuing modifications of the contract. Although we have taken the major responsibility for the semi-annual revisions of the contract, other faculty members have been increasingly more explicit in their evaluations and suggestions as their experience with the contract has expanded. Their confidence in this method of clinical evaluation has increased with each semester of use. We plan to continue in the refinement of the instrument and will develop materials for the use of other faculty groups who are interested in developing similar contracts. We are also designing a survey to attempt to discover how widespread is the use of contract grading in clinical areas. PMID- 6276345 TI - Viral hepatitis (adenovirus) in a California sea lion. AB - A juvenile California sea lion (Zalophus californianus) died 28 days after admission to the California Marine Mammal Center. Necropsy revealed hemorrhagic fluid in the stomach and hepatomegaly. Histologically, there was evidence of multifocal necrotizing hepatitis and acute suppurative bronchopneumonia. Amphophilic intranuclear inclusions were found, and electron microscopy revealed virions morphologically classifiable as adenovirus. PMID- 6276346 TI - Antibody response to an inactivated vaccine for rhinotracheitis, caliciviral disease, and panleukopenia in nondomestic felids. AB - The efficacy of an inactivated vaccine for the prevention of feline viral rhinotracheitis (FVR), feline caliciviral disease (FCVD), and feline panleukopenia (FPL) was tested in 27 nondomestic adult felids from 7 species. The vaccine was given IM at the standard domestic cat dose in 19 animals and double this dose in 8 others. The animals were vaccinated either 1, 2, or 3 times. Serum neutralization (SN) antibodies to FVR (mean SN titer, 23) developed in all 15 animals that were previously seronegative, and SN antibodies to FCVD (mean SN titer, 11) developed in 19 of 21 animals that were previously seronegative. There was no significant increase of SN antibody titers by doubling the vaccine dose or by administering a 3rd vaccination. The optimal response could be obtained by using the domestic cat vaccination protocol of a single dose given twice, 4 weeks apart. The critical evaluation of the SN antibody titer for FPL was complicated by preexisting titers to FPL from previous vaccinations, but in 23 animals the titers became higher, whereas they remained unchanged in only 4 animals. The persistence of the SN titers was evaluated 7 to 9 months later and found to be satisfactory for FVR (mean SN titer, 18) FCVD (mean SN titers, 43) and FPL (mean SN titer, 517). Enhanced persistence of titer could not be demonstrated by doubling the dose or administering a 3rd vaccination. PMID- 6276348 TI - Avian pox in blue-fronted Amazon parrots. AB - During a 1-month period at a quarantine station, an epornitic of avian pox occurred in blue-fronted Amazon parrots (Amazona aestiva). Clinical signs included conjunctivitis, blepharitis, and varying degrees of anorexia and respiratory distress. Lesions included periocular ulcerations and scabs and necrotic plaques in the oral cavity. Histologically, the lesions consisted of epithelial hyperplasia, secondary inflammatory changes, and eosinophilic inclusions which, by electron microscopy, were shown to contain poxvirus. When chicken embryos were inoculated with material from eyelid scabs and pharyngeal plaques, lesions of avian pox developed on the chorioallantoic membrane. The death rate of infected birds was high because of secondary bacterial and fungal infections, but uncomplicated cases were usually self-limiting. Periocular lesions also developed in 2 other species of psittacine birds housed in the same facility. PMID- 6276347 TI - Prevalence and sequelae of feline leukemia virus infection in laboratory cats. AB - Over a 4-year period, 1,683 pound-source cats received at a research institution were screened for feline leukemia virus (FeLV) infection, using an indirect fluorescent antibody test. Viremia was detected in 83 of the cats, for a prevalence of 4.9%. During this period, FeLV infection was detected in 5 kittens on a research project; lymphoma or anemia developed 6 to 17 months after the infections were detected. It was concluded that apparently healthy cats infected with FeLV may not be appropriate for some biomedical research projects. PMID- 6276349 TI - Fatal herpesvirus infection in patas monkeys and a black and white colobus monkey. AB - Fatal herpesvirus infections were diagnosed in 3 patas monkeys and 1 black and white colobus monkey over a 4-week period. Herpesvirus was isolated from 1 patas monkey and from the black and white colobus monkey. Both isolates had growth characteristics similar to Herpesvirus hominis and Herpesvirus simiae. The isolate from the colobus monkey antigenically appeared to be H simiae or H simiae like, whereas the isolate from the patas monkey could not be conclusively identified with the antisera used. All affected animals were housed in close proximity to rhesus monkeys, the carrier host of H simiae. PMID- 6276350 TI - Avian pox in pen-raised bobwhite quail. PMID- 6276351 TI - Chronic endocrine disorder associated with inclusion body hepatitis in a sulfur crested cockatoo. PMID- 6276352 TI - Cholangiocellular adenocarcinoma, leiomypma, and pheochromocytoma in a sea otter. PMID- 6276353 TI - Transmission of equine infectious anemia virus from horses without clinical signs of disease. AB - Twenty seven adult horses positive to the agar gel immunodiffusion (AGID) test for equine infectious anemia (EIA), but with no history of clinical EIA, were used in transfusion studies to determine whether infectious EIA virus was present in 1 to 5 ml of their blood. Of 27 recipients, 21 (78%) became AGID test-positive at an average of 24 days after inoculation. Two horses that were initially negative when screened were retested and found to carry infectious virus in 5-300 ml of whole blood; the other 4 horses were not retested. Horse flies (Tabanus fuscicostatus Hine) were unable to transmit EIA virus from 10 AGID test-positive donors with no history of clinical EIA, but virus was transmitted from a pony with artificially induced acute EIA and from a horse that had recovered from a clinical attack of EIA 9 months earlier. Histopathologic changes indicative of EIA were noted in all test-positive recipients. The most consistent lesion was paracortical lymphoid hyperplasia in the splenic lymph node. PMID- 6276354 TI - Detection of equine infectious anemia virus in a horse with an equivocal agar gel immunodiffusion test reaction. AB - A horse whose serum reacted equivocally in the agar gel immunodiffusion (AGID) test for equine infectious anemia was studied over a 3-year period. The horse remained afebrile and virus was detected in only 1 of 6 horse inoculation tests. The intensity of AGID test reactions increased temporarily following this evidence for virus. Although the AGID test reaction was equivocal and 5 of the 6 transmission attempts failed, the 1 successful transmission proved the horse was infected. PMID- 6276355 TI - In-vitro activity of SQ 26,776 against multiply-resistant enterobacteria- preliminary results. PMID- 6276356 TI - Recovery of eggs of two parasitic nematodes, Ascaris sp. and Trichuris sp.: Interlaboratory study. AB - An interlaboratory study was conducted to determine the effectiveness of the Nacconol ether centrifugation method for recovering parasitic nematode eggs from 3 contaminated products: a crop (cabbage), a sludge fertilizer (Milorganite), and a sewage effluent (Minneapolis). Six replicate samples for each of the 3 products were seeded with eggs at 3 different levels: 200 Ascaris suis and 8 Trichuris muris; 15 A.suis and 15 T.muris; 8 A.suis and 180 T.muris. Recovery was low for all samples except sewage effluent, in which recoveries greater than 100% in 2 samples resulted from the misidentification of arthropod eggs as Ascaris sp. The average mean percent recovery for the other samples was 22.53. Repeatability for replicate samples and reproducibility of results by individual laboratories were poor, and the method is not recommended for quantitative estimates of nematode egg contamination of foods and food-contact materials. However, the Nacconol ether centrifugation method can be used as an all-or-none test. (Only 13% of 1146 counts were falsely negative.) Of 69 samples, only 4 were falsely negative for A.suis eggs and only 1 was falsely negative for T.muris eggs in counts of 6 replicates. PMID- 6276358 TI - Partial purification and cleavage specificity of a site-specific endonuclease, SciNI, isolated from Spiroplasma citri. AB - A site-specific endonuclease, SciNI, has been partially purified from the plant pathogen Spiroplasma citri. The enzyme recognizes the sequence 5'-G-C-G-C-3' and cleaves between the first G and C. 3'-C-G-C-G-5' SciNI is an isoschizomer of HhaI, but generates DNA fragments with 5' rather than 3' single-stranded protrusions. PMID- 6276357 TI - Polypeptides encoded by the mer operon. AB - HgCl2-induced polypeptides synthesized by Escherichia coli minicells containing recombinant or natural HgR plasmids were labeled with [35S]methionine and analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. All plasmids examined encoded two heavily labeled, HgCl2-inducible polypeptides of 69,000 and 12,000 daltons. Most plasmids also encoded two additional HgCl2 inducible proteins in the 14,000- to 17,000-dalton range. Antiserum prepared against a purified mercuric ion reductase reacts with the 69,000-dalton polypeptide and a minor 66,000-dalton protein seen in several different HgR minicells. Recombinant plasmids constructed from portions of mer DNA from the IncFII plasmid NR1 were also analyzed in the minicell system. Five HgCl2 inducible polypeptides (69,000, 66,000, 15,100, 14,000, and 12,000 daltons) were synthesized in minicells carrying pRR130, a recombinant derivative containing the EcoRI-H and EcoRI-I restriction fragments of NR1. The EcoRI-H fragment of NR1 encodes the three small mer proteins of 15,100, 14,000, and 12,000 daltons and the amino-terminal 40,000 daltons of the mercuric ion reductase monomer. PMID- 6276359 TI - P1 transduction mapping of the trg locus in rac+ and rac strains of Escherichia coli K-12. AB - The trg locus, which had been located at min 31 in the cotransduction gap in the terminus region of the chromosome of Escherichia coli, has been mapped by transduction with bacteriophage P1. This locus exhibited no cotransduction with fnr when rac+ strains were used. If rac strains were used, which removed approximately 27 kilobase pairs of DNA, trg and fnr exhibited 8.2% cotransduction. Although this mapping of trg at min 31.1 considerably reduces the size of the cotransduction gap, trg exhibited no cotransduction with a Tn10 insertion located on the other side of the gap at min 34.2. PMID- 6276360 TI - The herpes simplex virus thymidine kinase gene is not transcribed in Saccharomyces cerevisiae. AB - The herpes simplex virus thymidine kinase gene has been cloned into a chimeric yeast plasmid cloning vehicle and transformed into appropriate yeast strains. Plasmids carrying the herpes simplex virus thymidine kinase gene can be propagated as autonomously replicating plasmids, but no RNA specific to the thymidine kinase coding sequence was detected. PMID- 6276361 TI - Evidence for an internal promoter preceding tufA in the str operon of Escherichia coli. AB - We constructed plasmids carrying tufA from which the major promoter for the rpsL rpsG-fus-tufA operon (also called the str operon) had been removed. These plasmids continued to express tufA, as judged by the ability to complement mocimycin resistance and by electrophoretic analysis of synthesized proteins. Tn5 transpositions into fus, the gene for elongation factor G, which lies immediately on the 5' side of tufA, failed to obstruct the expression of tufA. The subcloning of a 2,000-base-pair PstI-SmaI DNA fragment (containing the intercistronic region between tufA and fus, the distal portion of fus, and the proximal portion of tufA) next to promoterless tetracycline resistance genes (tet) yielded a plasmid that was capable of bestowing resistance to 12 microgram of tetracycline per ml. The removal of an EcoRI fragment that lies within fus destroyed the ability of the 2,000-base-pair PstI-SmaI fragment to promote the transcription of tet. These data indicate that, in addition to the operon's major promoter rpsLp, there is an internal promoter, tufAp, which can be used for the transcription of tufA, tufAp probably lies within fus, about 50 base pairs upstream from its 3' end and 120 base pairs from the start codon of tufA. The relative activities of tufB and of tufA-from-tufAp were estimated by a comparison of beta-galactosidase activities of almost identical EF-Tu-beta-galactosidase protein fusions; they were approximately equal. PMID- 6276362 TI - Cloning and endonuclease restriction analysis of uidA and uidR genes in Escherichia coli K-12: determination of transcription direction for the uidA gene. AB - The two genes of the Escherichia coli K-12 uid region (the structural gene uidA and the regulatory gene uidR) were isolated on a ColE1-uid hybrid plasmid from the bank of Clarke and Carbon. We made a restriction map of this region and correlated it with the genetic map by subcloning the uid restriction fragments into plasmids pBR322, pBR325, and pACYC177. In these plasmids, amplification of the products of the uidA and uidR genes occurred. The enzyme coded for by uidA was identified by polyacrylamide gel electrophoresis of crude extracts from strains containing the uidA plasmid. A 1-megadalton EcoRI-BamHI segment contained the uidAo operator, and the direction of transcription of the uidA gene was determined. The restriction analysis also suggested that the order of the loci in this region is manA, uidA, uidAo, uidR. PMID- 6276363 TI - Uptake and fate of bacteriophage phi W-14 DNA in competent Bacillus subtilis. AB - Phage phi W-14 DNA (in which one-half of the thymine residues are replaced by alpha-putrescinyl thymine) was taken up by competent Bacillus subtilis cells at a rate threefold higher than the rate of homologous DNA uptake. In contrast to other types of heterologous DNA, the amount of phi W-14 DNA taken up in 15 min exceeded the amount of homologous DNA taken up by a factor of two to three, as measured in terms of acid-precipitable material. The amount of phi W-14 DNA taken up was even greater than this analysis indicated if allowance was made for the fact that phi W-14 DNA was degraded more rapidly after uptake than homologous DNA. Competition experiments showed that the affinity of phi W-14 DNA for homologous DNA receptors was lower than the affinity of homologous DNA and was similar to the affinities of other types of heterologous DNA. The more rapid and more extensive uptake of phi W-14 DNA appeared to occur via receptors other than the receptors for homologous DNA, and these receptors (like those for homologous DNA) were an intrinsic property of competent cells. Uptake of phi W-14 DNA was affected by temperature, azide, EDTA, and chloramphenicol, as was uptake of homologous DNA. This was consistent with entry of both DNAs by means of active transport. After uptake, undegraded phi W-14 [3H]DNA was found in the cells in a single-stranded form, whereas a portion of the label was associated with recipient DNA, presumably as a result of incorporation of monomers resulting from degradation. Acetylation of the amino groups of the putrescine side chains in phi W-14 DNA decreased the affinity of this DNA for its receptors without affecting its ability to compete with homologous DNA. PMID- 6276364 TI - Transposon mutagenesis in Caulobacter crescentus. AB - Transposons Tn5 (Km) and Tn7 (Tp and Sm) were transferred to Caulobacter crescentus via P-type antibiotic resistance factors. Transposition was demonstrated by the isolation of chromosomal insertions of each transposon. With C. crescentus strains harboring RP4 aphA::Tn7, the introduction of a wild-type RP4 resulted in the loss of the resident plasmid. Simultaneous selection for Kmr and Smr yielded colonies with chromosomal insertions of Tn7. Examination of over 10,000 chromosomal insertions of Tn7 indicated no auxotrophic or motility mutants. Thus, Tn7 appears to have a high specificity of insertion in C. crescentus. The Mu-containing plasmid pJB4JI transferred Tn5 to C. crescentus, but the plasmid was not maintained. Control experiments showed that recovery of Mu-containing plasmids occurred at very low frequencies in C. crescentus and that the plasmids which were recovered had undergone extensive deletion of plasmid DNA. Presumably, some part of the Mu genome was not tolerated by C. crescentus. The instability of the Mu-containing plasmids makes them excellent vectors for the introduction of transposons, and we have used pJB4JI to isolated chromosomal insertions of Tn5. When several thousand of these insertion mutants were examined, we found auxotrophic and motility mutants at frequencies of 1 and 2%, respectively. These results indicate that Tn5 had a low specificity of insertion in C. crescentus and therefore would be a useful mutagen for obtaining a variety of mutant phenotypes. PMID- 6276365 TI - Cloning of phoE, the structural gene for the Escherichia coli phosphate limitation-inducible outer membrane pore protein. AB - The hybrid plasmid pLC44-11 from the Clarke and Carbon collection, which was known to carry the proA gene, was shown also to contain the phoE gene. In vitro recombination techniques were used to subclone a 4.9-kilobase-pair DNA fragment of pLC44-11 into the plasmid vectors pACYC184 and pBR322. Expression of this fragment in a minicell system showed that it codes for the PhoE protein and for polypeptides with apparent molecular weights of 47,000 and 17,000. These results supply definite proof for the earlier supposition that the phoE gene is the structural gene for the outer membrane PhoE protein. Overproduction of the PhoE protein in a phoS strain resulted in reduced amounts of OmpF and LamB proteins. PMID- 6276366 TI - Physical mapping and cloning of bacteriophage T4 anti-restriction endonuclease gene. AB - We have proposed that the ability of T4 to produce non-glucosylated progeny after a single cycle of growth on a galU rglA rglB+ mutant of Escherichia coli is due to the initiation of the rglB+ function by a phage-coded, anti-restriction endonuclease protein. Based on this hypothesis, we screened T4 deletion mutants for failure to give a burst in this host. The absence of an arn gene in phage mutants lacking the 55.5- to 58.4-kilobase region is verified by their inability to protect secondary infecting non-glucosylated phage from rglB-controlled cleavage. A functional arn gene was cloned on plasmid pBR325, and the 0.8 kilobase insert DNA was shown to be homologous to the DNA missing in the arn deletion phage. PMID- 6276367 TI - Spread of a "Pseudomonas-specific" beta-lactamase to plasmids of enterobacteria. AB - Eleven isolates including Escherichia coli, Salmonella enteritidis, and Shigella sonnei, obtained in Brazil, Hong Kong, Indonesia, Thailand, and the United States, were found to produce beta-lactamase of the PSE-1 type, which was previously considered to be Pseudomonas specific. The enterobacterial strains produced a beta-lactamase with the same isoelectric point, immunological reactions, and substrate profile as those of the prototype PSE-1 enzyme determined by Pseudomonas plasmid RPL11. The producer strains were resistant to multiple antibiotics, and all contained plasmids, ranging in size from 37 x 10(6) to 130 x 10(6), that belonged to at last six incompatibility groups. Plasmids of IncH2 and IncFIme were shown to contain 8 x 10(6)-molecular-weight transposons Tn1401 and Tn1402 that encoded PSE-1 beta-lactamase production, resistance to streptomycin and spectinomycin via AAD(3"), and resistance to sulfonamide. PSE-1 beta-lactamase was not Pseudomonas specific and appeared to have spread among plasmids found in enterobacteria by transposition. PMID- 6276368 TI - Mobilization of nonconjugative antibiotic resistance plasmids in Haemophilus ducreyi. AB - A clinical isolate of Haemophilus ducreyi was found to harbor three plasmids: a 23.5-megadalton (Mdal) phenotypically cryptic plasmid, a 7.0-Mdal ampicillin resistance plasmid, and a 4.0-Mdal sulfonamide resistance plasmid. The two smaller plasmids were transferable by conjugation to Haemophilus recipients, but only if the donor cell harbored the 23.5-Mdal plasmid as well, indicating that this large plasmid had mobilizing capabilities. Transfer was also possible to Escherichia coli recipients. Haemophilus influenzae transconjugants which had acquired both the 23.5-Mdal plasmid and one of the R-plasmids could subsequently retransfer the R-plasmid to other Haemophilus recipients at higher frequencies. A derivative of the 23.5 Mdal plasmid was isolated which was shown by restriction endonuclease analysis to contain an ampicillin resistance transposon and to have retained its conjugative ability. PMID- 6276370 TI - Extracellular exonuclease as a stage 0 biochemical marker in Bacillus subtilis sporulation. AB - Calcium-dependent exonuclease activity was produced by sporulating cells of Bacillus subtilis 168. Nuclease activity was released into the culture medium at approximately the same time as sporulation proteases, and production of these enzymes was tied to DNA replication. Results suggest that nuclease production is a function of the spo0H locus. PMID- 6276369 TI - Pheromone produced by the myxobacterium Stigmatella aurantiaca. AB - An extracellular, diffusible signaling molecule (pheromone) was produced by Stigmatella aurantiaca during fruiting body formation. The pheromone decreased the aggregation period in both the light and the dark and substituted for light in stimulating the maturation of aggregates into fruiting bodies. The cells were more sensitive to lower concentrations of pheromone in the light than in the dark, possibly explaining the stimulation of aggregation and fruiting body formation by light. The pheromone also interacted cooperatively with GMP to shorten the aggregation period. The pheromone behaved chemically as a low molecular-weight lipid. PMID- 6276371 TI - [Ultrastructural study of enamel and dentin in regional odontodysplasia]. PMID- 6276373 TI - Inactivation of yeast fructose-1,6-bisphosphatase. In vivo phosphorylation of the enzyme. AB - Incorporation of 32P into yeast fructose-1,6-bisphosphatase (EC 3.1.3.11) was observed after addition of glucose to a cell suspension incubated with (32P)orthophosphoric acid. The 32P counts were coincident with the enzyme band when immunoprecipitates were subjected to sodium dodecyl sulfate disc gel electrophoresis. The incorporation of phosphate was associated with a decrease in enzyme activity. Approximately 1 mol of phosphate was incorporated/mol of enzyme. The phosphate is bound to the enzyme in a phosphoester linkage with a serine residue. Release of 32P accompanying enzyme reactivation was observed both in vivo and in cell-free extracts. PMID- 6276372 TI - [Effect of vitamin D3 and vitamin D3 sulfate on dental and bone tissues in the pig]. AB - The repairing effects of vit. D3 and vit. D3 sulfate upon the hard tissues of vit. D3 deficient pigs have been studied using SEM and electron microprobe analyses. It was demonstrated that a vit. D3 therapy could balance lesions induced by vit. D3 deficiency. The effects of vit. D3 sulfate were not so clear. Results obtained were varying, sometimes similar to those obtained with vit. D3 but not always statistically significant. The rate of mineralization of dentine and bone was higher in animals having received vit. D3 and vit. D3 sulfate compared to the deficient group. In dentine and bone, the Ca/P ratio, thus the crystalline phases, wee modified, which was not the case for enamel and acellular cementum. It was shown that dentine and bone, which both include cells and cell processes in the calcified matrix, undergo similar modifications in the course of vit. D3 deficiencies and their therapy. On the contrary, enamel and acellular cementum in vit. D3 deficiency presented a lower rate of mineralization without any structural defects or modifications in Ca, P, Mg and F- ratios. Repair of lesions due to vit. D3 deficiencies by vit. D3 and, to a lower degree, by vit. D3 sulfate, concern, in order, bone, dentine, cementum and very little enamel. PMID- 6276374 TI - Demonstration of the feasibility of observing nuclear magnetic resonance signals of 77Se covalently attached to proteins. AB - Previous 77Se NMR relaxation time studies established the utility of 77Se NMR spectroscopy in studying low molecular weight (less than 500) selenium-containing molecules. Since the spin rotation and chemical shift anisotrophy mechanisms contributed significantly to the 77Se spin-lattice relaxation in these compounds, it was questionable as to whether the latter mechanism would be efficient enough to enable 77Se resonances to be observed in a reasonable period in high molecular weight selenobiomolecules. Thus, to address this problem, disulfide bonds of ribonuclease-A and lysozyme were reductively cleaved under denaturing conditions, and the resulting 7-8 sulfhydryl groups were treated with a new sulfhydryl group reagent containing selenium, 6,6'-diselenobis(3-nitrobenzoic acid), to give proteins containing covalently attached selenium in the form of selenenyl sulfides. The observation of high resolution 77Se NMR spectra of these proteins under denaturing conditions was accomplished. Five to six 77Se NMR resonances, which fell in a chemical shift range of 14-15 ppm, were observed for each protein and are compared to the chemical shifts of several model selenenyl sulfides derived from cysteine. PMID- 6276375 TI - The role of the membrane in the regulation of activity of microsomal glucose-6 phosphatase. AB - The factors regulating glucose-6-phosphatase (EC 3.1.3.9) activity and substrate specificity in hepatic microsomes were studied by determining the rate-limiting reaction for the hydrolysis of glucose-6-P, and by examining the effect of detergent activation on phosphotransferase activity. Examination of the pre steady state kinetics of glucose-6-phosphatase revealed that the steady state rate is determined by the rate of hydrolysis of the enzyme-P intermediate. Treatment of the enzyme with detergent does not alter the extent of the rapid release of glucose per mg of protein, but activates the steady state rate of catalytic turnover. Specificity of the enzyme was evaluated by comparing the effects of mannose and glucose as phosphate acceptors in the phosphotransferase reaction catalyzed by glucose-6-phosphatase. Untreated glucose-6-phosphatase discriminates against mannose as compared with glucose in that mannose and glucose bind to the enzyme-P intermediate of untreated enzyme, but mannose is not an acceptor of Pi. Mannose is an acceptor, however, after treatment of microsomes with detergent. These data cannot be explained in terms of the currently accepted "compartmentation" model for the regulation of glucose-6-phosphatase. The detergent-induced changes in kinetic properties appear to reflect alterations in the intrinsic characteristics of glucose-6-phosphatase, which could result from interaction with its membrane environment. PMID- 6276376 TI - Phosphorylation by guanosine 3':5'-monophosphate-dependent protein kinase of synthetic peptide analogs of a site phosphorylated in histone H2B. AB - Analogs of a synthetic heptapeptide substrate corresponding to the sequence around a phosphorylation site in histone H2B were used to assess the substrate specificity of cGMP-dependent protein kinase. cGMP-dependent kinase phosphorylated the oligopeptide Arg-Lys-Arg-Ser32-Arg-Lys-Glu with favorable kinetic parameters as compared to those for cAMP-dependent kinase (Glass, D. B., and Krebs, E. G. (1979) J. Biol. Chem. 254, 9728-9738). The contribution of each amino acid to the ability of the peptide to be phosphorylated by cGMP-dependent or cAMP-dependent kinase was studied by replacement of individual residues and evaluation of the kinetic constants of the substituted peptides. Peptides containing acetylated lysine residues or nitroarginine residues were poor substrates for both kinases. Substitution of either arginine 29 or lysine 30 with alanine increased the Km values and decreased the Vmax values for both kinases. Substitution of lysine 34 with alanine increased the Vmax values for both kinases but did not affect the Km values for either enzyme. Substitution of the phosphorylatable serine with a threonine residue greatly depressed the Vmax for both kinases. Peptides in which arginine 31 or arginine 33 were replaced by an alanine residue revealed several apparent differences in the specificity requirements between cGMP-dependent and cAMP-dependent kinases. PMID- 6276377 TI - Endocytosis and degradation mediated by the asialoglycoprotein receptor in isolated rat hepatocytes. PMID- 6276378 TI - Function and regulation of methionine5-enkephalin and its receptors in murine neuroblastoma cells. AB - A homogeneous class of enkephalin receptors found in murine neuroblastoma clone N1E-115 (Chang, K.-J., and Cuatrecasas, P. (1979) J. Biol. Chem. 254, 2610-2618) has been confirmed using a centrifugation assay employing cellular membranes. In intact N1E-115 cells, synthetic methionine5-enkephalin inhibited prostaglandin E1 induced intracellular cyclic AMP formation in a naloxone-sensitive manner. Upon demonstrating intracellular methionine5-enkephalin immunocytochemically (Knodel, E., and Richelson, E. (1980) Brain Res. 197, 565-570), analyses of crude N1E-115 extract were made by radioimmunoassay or opiate receptor binding assay following fractionation by molecular sieve chromatography and high pressure liquid chromatography on a mu-Bondapak C18 column. Extracted methionine5-enkephalin immunoreactive material behaved similarly to synthetic methionie5-enkephalin in these analyses. Growth curve studies of the N1E-115 cells indicated that the quantity of methionine5-enkephalin immunoreactive material synthesized per milligram of cellular protein and the maximum number of enkephalin receptor sites per milligram of membrane protein increased as the cells progressed from logarithmic to stationary phase, with no change in the apparent affinity of the enkephalin receptors for [3H]methionine5-enkephalin. These data suggest that adrenergic clone N1E-115 has functional methionine5-enkephalin membrane receptors, that this clone synthesizes methionine5-enkephalin, and that both the enkephalin receptor number and the content of stored methionine5-enkephalin are regulated with respect to cell division. PMID- 6276379 TI - Direct inhibition of rat testicular androgen biosynthesis by arginine vasotocin. Studies on mechanisms of action. PMID- 6276380 TI - The refined crystal structure of ribonuclease A at 2.0 A resolution. AB - This paper describes the structure of bovine pancreatic ribonuclease A, refined by a restrained parameter least squares procedure at 2.0 A resolution, and rebuilt using computer graphics. The final agreement factor (formula see text) is 0.159. The positions of the 951 main chain atoms have been determined with an estimated accuracy of 0.17 A. In addition, the model includes a phosphate group in the active site and 176 waters, many of them with partial occupancy. The bond lengths in the refined structure of RNase A differ from the ideal values by an overall root mean square deviation of 0.022 A; the corresponding value for angle distances is 0.06 A. The root mean square deviation of planar atoms from ideality is 0.017 A, and root mean square deviation of the peptide torsion angles from 180 degrees is 3.4 degrees. The model is in good agreement with the final difference Fourier maps. Two active site histidines, His 12 and His 119, form hydrogen bonds to the phosphate ion. His 119 is also hydrogen bonded to the carboxyl of ASp 121 and His 12 to the carbonyl of Thr 45. The structure of the RNase A is very similar to that of RNase S, particularly in the active site region. The root mean square discrepancy of all atoms from residues 1 to 16 and 24 to 123 is 1.06 A and the root mean square discrepancy for the active site region is 0.6 A. PMID- 6276381 TI - Chemical and spectral properties of carbon monoxide: methylene blue oxidoreductase. The molybdenum-containing iron-sulfur flavoprotein from Pseudomonas carboxydovorans. AB - Carbon monoxide:methylene blue oxidoreductase, the key enzyme of CO-oxidation in energy metabolism of the carboxydobacterium Pseudomonas carboxydovorans, has been isolated in good yield and purity and found to contain FAD, molybdenum, iron, and labile sulfide in the ratio of 1:1:4:4. The enzyme is, therefore, a new molybdenum-containing iron-sulfur flavoprotein, exhibiting chemical and spectral properties quite similar to those of xanthine oxidase. Analytical data on the spectral characteristics of the enzyme in the oxidized and various reduced states are presented. Carbon monoxide:methylene blue oxidoreductase turned out to be photoreducible in the presence of EDTA and urea and was subject to reoxidation by air oxygen; no flavoprotein semiquinone was formed. Unphysiological electron acceptors, e.g. methylene blue, were used as oxidizing substrates whereas NAD or NADP turned out to be ineffective. Methylene blue reduction with CO was not affected by the presence of allopurinol, and carbon monoxide:methylene blue oxidoreductase was not able to catalyze the reduction of methylene blue with xanthine, adenine, or aldehydes. CO was the only reducing substrate used by the enzyme. Carbon monoxide:methylene blue oxidoreductase formed no sulfite adduct, and the reactivity with ferricyanide or cytochrome c was significant but slow. As known for other molybdenum hydroxylases, carbon monoxide:methylene blue oxidoreductase was rapidly inactivated by methanol, but the enzyme exhibited no ability to catalyze the oxidation of NADH with methylene blue, and NAD was not able to overcome methanol inhibition. PMID- 6276382 TI - Inhibitors of specific aminoacyl-tRNA synthetases prevent thyrotropin-induced desensitization in cultured human thyroid cells. PMID- 6276383 TI - Evidence for the inorganic nature of the cyanolyzable sulfur of molybdenum hydroxylases. AB - Activation of the desulfo forms of milk xanthine oxidase, chicken liver xanthine dehydrogenase, and aldehyde oxidase with S2- is greatly facilitated in the presence of reducing agents. Upon anaerobic incubation with 1 mM S2- and 1 mM dithionite, desulfo xanthine oxidase and chicken liver xanthine dehydrogenase prepared by cyanide treatment of active enzymes, are activated to the specific activity predicted by their molybdenum content. Routine preparations containing desulfo molecules are also similarly activated to the extent predicted. Cyanide inactivated chicken liver xanthine dehydrogenase was reconstituted with 35S2- in the presence of dithionite. 85% of enzyme-bound radioactivity was shown to be in the form of cyanolyzable sulfur, by comparison of enzyme activity, bound radioactivity, and 35SCN- yields from exposure of labeled enzyme to cyanide. This radiolabeled enzyme allowed the determination of the following. 1) The cyanolyzable sulfur is largely removed from the polypeptide by incubation at 37 degrees C for one hour in 1% sodium dodecyl sulfate, pH 7, or for 15 min in 6 M guanidinium chloride, pH 6.2. 2) The cyanolyzable sulfur is "acid labile." [35S]Methylene blue is formed in the theoretical quantity from oxidized or substrate-reduced enzyme under the standard conditions for labile sulfur analysis by the methylene blue method. These data strongly support the conclusion that the cyanolyzable sulfur is a terminal sulfur ligand of the Mo atom, and is not part of an organic moiety. PMID- 6276384 TI - Thermodynamics of triiodothyronine nuclear receptor interaction. Role of hydrophobicity in triiodothyronine binding to its receptor. AB - We have investigated the thermodynamic properties of triiodothyronine (T3) solubilized hepatic nuclear receptor interaction. These studies revealed that the equilibrium constant was markedly dependent on temperature and van't Hoff plots revealed a significant deviation from linearity. Both enthalpy and entropy changes decreased with increasing temperature such that at 37 degrees C both were negative. (delta H0 -27.1 kcal/mol; delta S0, -45.1 cal/mol.deg; and the heat capacity delta Cp0 (25 degrees C), -759 cal/mol.deg). Several T3 analogues revealed similar thermodynamic characteristics. Our finding that the T3-receptor interaction is characterized by a negative heat capacity is compatible with the previous proposal of hydrophobic bonding by Jorgensen. The large negative entropy change at 37 degrees C for T3-receptor reaction is in contrast to the more positive entropy values for several T3-plasma protein reactions. One possible explanation of this difference in entropy values between the nuclear receptor and plasma proteins is that T3 induces a conformational change in the receptor, a concept supported by previous data from our laboratory indicating that the occupation of the T3 receptor results in an alteration in the chromatographic migration of the receptor. PMID- 6276385 TI - Regulation of sphingomyelin long chain base synthesis in human fibroblasts in culture. Role of lipoproteins and the low density lipoprotein receptor. AB - We have studied regulation of synthesis of long chain bases in human fibroblasts using 3 different radioactive precursors and 2 different hydrolysis and separation procedures. Serum and low density lipoproteins inhibited synthesis. Inhibition of long chain base synthesis by various concentrations of low density lipoproteins paralleled inhibition of cholesterol synthesis. This inhibition was dependent on the low density lipoprotein receptor pathway since fibroblasts from homozygous familial hypercholesterolemic patients did not show the inhibition observed with normal fibroblasts. Incorporation of precursor palmitate into free or total long chain bases was inhibited by low density lipoproteins to the same extent as incorporation into sphingomyelin long chain bases. We thus propose that an enzyme in the pathway leading to sphinganine synthesis, probably palmitoyl CoA:L-serine C-palmitoyltransferase (decarboxylating) EC 2.3.1.50, is regulated by low density lipoproteins. PMID- 6276386 TI - Cyclic AMP-dependent and cyclic AMP-independent antagonism of insulin activation of cardiac glycogen synthase. AB - The hormonal regulation of glycogen synthase has been studied with isolated perfused hearts that were depleted of 85% of their endogenous glycogen. Glycogen depletion alone promoted a 3-fold activation of glycogen synthase and magnified by 3-fold the response to insulin. Glycogen depletion also facilitated the detection of epinephrine-promoted glycogen synthase inactivation. Hormonal effects on glycogen synthase have been correlated with changes in phosphorylase, phosphorylase kinase, and tissue cAMP levels. Insulin activation of glycogen synthase was observed within 90 s of hormone addition and was maximal by 4 min. A half-maximum effect was obtained at an insulin concentration of 100 microunits/ml. Insulin-dependent activation is reversed by beta-adrenergic agonists, alpha-adrenergic agonists, and glucagon. Each promote the same degree of inactivation and the maximum extent of inactivation produced by each is independent of whether or not the tissue has been stimulated with insulin. beta Adrenergic agonists and glucagon act via cAMP, alpha-agonists most likely act via intracellular Ca2+ translocation, and insulin action would appear to be independent of either cAMP or Ca2+. The action of epinephrine on cardiac glycogen synthase is mediated by interaction with both alpha- and beta-receptors. As indicated by dose-response curves, receptor occupancy of each occurs to an almost equal extent at suboptimal epinephrine concentrations. Regulation of cardiac glycogen synthase by epinephrine thus is mediated by two second messenger systems which converge to produce the end physiological response. PMID- 6276387 TI - Yeast gene TRP5: structure, function, regulation. AB - The nucleotide sequence of the yeast gene TRP5 and its 5' and 3' flanking regions was determined. The deduced coding sequence for tryptophan synthase contains 2,127 base pairs. The protein chain has a calculated molecular weight of 76,544. Yeast tryptophan synthase, a bifunctional protein, has a primary structure which corresponds to an Escherichia coli tryptophan synthase alpha chain-beta chain fusion. An NH2-terminal 239 amino acid segment of yeast tryptophan synthase is homologous with E. coli tryptophan synthase alpha subunit, while a distal 389 amino acid residue segment is homologous to the E. coli tryptophan synthase beta chain. This order of segments of the yeast enzyme is the reverse of the chromosomal order characteristic of all prokaryotes that have been examined. The two segments are joined by a connecting region of 28 residues in the yeast enzyme which is not homologous to either the alpha or beta chains of the bacterial enzyme. A portion of the connecting region of yeast tryptophan synthase exhibits nucleotide sequence similarity to the 3' terminus of E. coli trpC and the trpC trpB intercistronic region. Active site cysteine, histidine, and lysine residues in the beta 2 subunit of E. coli tryptophan synthase are conserved in the yeast enzyme. Also conserved in the yeast enzyme are 6/8 amino acid residues having an important role in maintaining the structure and function of the E. coli tryptophan synthase alpha subunit. S1 nuclease mapping was used to identify three major mRNA transcripts with different 5' termini. Potential T-A-T-A sites for transcription initiation were identified, as well as other sequences that occur frequently in yeast genes. A 5' flanking region of TRP5 was shown by DNA/DNA hybridization to be present in multiple copies in the yeast genome. TRP5 mRNA levels, measured by RNA/DNA hybridization, increased 2- to 7-fold in response to starvation for either tryptophan or histidine, indicating transcriptional regulation. PMID- 6276388 TI - Definition of 5' and 3' structural boundaries of the chromatin domain containing the ovalbumin multigene family. AB - Hen oviduct nuclei were subjected to pancreatic DNase I treatment under conditions known to preferentially degrade transcriptionally active genes (Weintraub, H., and Groudine, M. (1976) Science (Wash. D. C.) 93, 848-856). The ovalbumin gene, its structurally related genes, X and Y, and the spacer and flanking DNA were all found to exist in a DNase I-sensitive configuration. The DNase I-sensitive region was extended more than 20 kilobases beyond the 5' end of the X gene and approximately an equal distance beyond the 3' end of the ovalbumin gene before it became DNase I-resistant. The transition from a DNase I-sensitive to a -resistant conformation in oviduct chromatin occurred in a gradient fashion with 10 kilobases of DNA. Thus, ovalbumin and its related genes, X and Y, exist in a 100-kilobase DNase-sensitive domain in the oviduct tissue. In contrast, the entire domain was resistant to DNase I in spleen, liver, and erythrocyte nuclei. When the transcription of ovalbumin, X, and Y genes was eliminated by the withdrawal of hormone from estrogen-stimulated chicks, the entire domain remained in a DNase I-sensitive configuration. We conclude that DNase I-sensitive domains may provide the structural capability for gene expression and appear to be a result of the differentiation process since they are cell-specific and contain potentially expressible genes of that cell type. Repetitive sequences within this domain have been mapped and the possible relationship of these repetitive sequences to the DNase I-sensitive structure is discussed. PMID- 6276389 TI - Evolution of human immunoglobulin kappa J region genes. AB - Immunoglobulin kappa chain variable region genes are assembled from two discontinuous DNA segments, a V and a J gene. The J region genes, in addition to encoding amino acid positions 96-108 of the kappa polypeptide chain, also provide sequences required for both DNA and RNA splicing reactions. For purposes of evolutionary comparison and to establish the complexity of the kappa J region locus in man, we have determined an approximately 3000 basepair nucleotide sequence in a cloned human DNA fragment that encodes the germline distinct J region segments. Significant blocks of homology have been tightly maintained between this region and an analogous segment of the mouse genome. In particular, the short sequences, GGTTTTTGT and CACTGTG, thought to be involved in V-J recombination, are the most highly conserved regions (97% homology). In addition, from heteroduplex data and computer analysis of the nucleotide sequences, it is clear that the mouse J3 sequence, a pseudogene, is not present in the human cluster. This can be explained by a duplication event in the mouse J region gene cluster that may have been the result of unequal crossing over between homologous chromosomes. PMID- 6276390 TI - A native 170,000 epidermal growth factor receptor-kinase complex from shed plasma membrane vesicles. AB - A method is presented for the preparation of a "native" epidermal growth factor (EGF) receptor-kinase complex of molecular weight 170,000 from A-431 cells. Although this receptor complex is capable of binding EGF, noncovalently, in quantities similar to the previously isolated 150,000 complex (Cohen, S., Carpenter, G., and King, L., Jr. (1980) J. Biol. Chem. 255, 4834-4842), the 170,000 preparation has 5 to 10 times the intrinsic kinase activity (autophosphorylation). However, the 170,000 kinase activity toward other proteins is lower than that of the 150,000 preparation. Both the 170,000 and 150,000 kinase activities are enhanced by EGF. The 170,000 and 150,000 proteins are also capable of forming covalent linkages to 125I-EGF, and each is precipitated by antisera directed against the 170,000 protein. We suggest the 150,000 protein is a proteolytic degradation product of the 170,000 protein. The EGF-enhanced kinase activity of the 170,000 preparation remains associated with the 125I-EGF-binding activity following EGF affinity chromatography, electrophoresis in nondenaturing gels, or immunoprecipitation with antisera directed against the sodium dodecyl sulfate (SDS) gel-purified 170,000 protein. These results indicate that the receptor, kinase, and substrate domains are linked, possibly covalently. PMID- 6276391 TI - Molecular interaction between simian virus 40 DNA and Escherichia coli RNA polymerase. Mapping of the initiation sites on supercoiled and linear DNA. AB - A detailed map of the initiation sites recognized by the Escherichia coli RNA polymerase (holoenzyme) on simian virus 40 (SV40) DNA has been constructed. An approach is described in order to compare the sites recognized on supercoiled as well as relaxed linear DNA. For localization of the initiation sites on supercoiled DNA, RNA polymerase-SV40 DNA Form I binary complexes are stabilized by incubation with three ribonucleoside triphosphates. The stable initiation complexes are cleaved with single cut restriction endonucleases in order to generate termination sites at precisely defined locations. Elongation of the RNA chains up to the cleavage site produces a defined set of discrete RNA species, whose size can be determined with accuracy by agarose gel electrophoresis following glyoxal denaturation. Each transcript is oriented by following two different experimental approaches. 1) The DNA of an SV40 deletion mutant is used as a template in comparison with that of wild type SV40; and 2) the RNAs synthesized after cleavage with different single cut restriction endonucleases are compared. Determination of the length and polarity of these RNAs allows us to accurately position the initiation sites on the physical map of the SV40 genome. A similar analysis was conducted for the mapping of the initiation sites recognized on precut linear SV40 DNA. Comparison of the two templates (supercoiled and linear) reveals that modification of the DNA conformation affects selection by the enzyme of certain promoters. Although most of the promoters are common to both templates, four are specifically recognized on superhelical DNA, while two others are used exclusively on linear SV40 DNA. PMID- 6276392 TI - Optimal conditions for post-translational uptake of proteins by isolated chloroplasts. In vitro synthesis and transport of plastocyanin, ferredoxin-NADP+ oxidoreductase, and fructose-1,6-bisphosphatase. AB - Many polypeptides translated in the cytosol enter the chloroplast where they assemble into macromolecular complexes. The transport of these polypeptides into the plastid can be examined in vitro by mixing isolated chloroplasts with pea poly(A) RNA translation products. Following optimization of both translation in the wheat germ system and the conditions during in vitro uptake, we observe the post-translational transport of over 100 polypeptides; many remain in the soluble phase of the organelle while others integrate into the thylakoid membranes. Most products transported in vitro co-migrate with in vivo products on sodium dodecyl sulfate-polyacrylamide gels. Furthermore, with the improved conditions, we demonstrate the transport of plastocyanin, ferredoxin-NADP+ oxidoreductase, and fructose-1,6-bisphosphatase into isolated plastids. While we have not been able to detect any cell-free translation product that is immunologically related to fructose-1,6-bisphosphatase, both plastocyanin and ferredoxin-NADP+ oxidoreductase are synthesized as precursors in vitro. These precursors are imported into the organelle where they are processed to the size of their mature counterparts. As determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the molecular weight of the precursor to plastocyanin is 15,000 larger than the mature product and the precursor to ferredoxin-NADP+ oxidoreductase is 8,000 larger than the mature product. PMID- 6276393 TI - The cytochrome c oxidase of Paracoccus denitrificans pumps protons in a reconstituted system. AB - The purified two-subunit cytochrome c oxidase of Paracoccus denitrificans was reconstituted into phospholipid vesicles having a high internal buffering capacity and exhibiting a respiratory control index greater than 6.6. With these proteoliposomes, pH changes of the suspending medium were monitored in response to reductant pulses in the presence of valinomycin and potassium. When reduced cytochrome c was added to allow for a limited number of turnovers (2-12), a net acidification of the extravesicular space could be observed. This apparent proton ejection by the vesicles was abolished by inhibition of the oxidase with azide, by bypassing the oxidase with ferricyanide, or by preventing charge compensation by omitting valinomycin. Addition of uncoupler led to an alkalinization, rather than an acidification, of the extravesicular space in response to reduced cytochrome c. We thus conclude that cytochrome c oxidase of P. denitrificans is a proton pump. Under the conditions described here, an apparent stoichiometry of 0.6 proton ejected/electron was obtained by extrapolation to zero turnovers. PMID- 6276394 TI - Hepatic glucagon-receptor complexes lose sensitivity to the dissociating effect of GTP. PMID- 6276395 TI - Dynamic interactions of CO with a3Fe and CuB in cytochrome c oxidase in beef heart mitochondria studied by Fourier transform infrared spectroscopy at low temperatures. AB - Carbon monoxide bound to cytochrome c oxidase has been observed by Fourier transform infrared spectroscopy between 10 K and 280 K in the dark and during and after continuous photolysis. CO bound to a3Fe absorbs near 1963 cm-1, with minor bands at lower frequencies. Photolysis at low temperatures transfers CO to CuB, with the major component near 2062 cm-1 and a minor one near 2043 cm-1. Vibrational absorptions are assigned by comparison with heme and copper carbonyls, by frequency dependence of all bands on the isotopic mass of CO, and by similar behavior of major and minor components with photolysis and relaxation kinetics as a function of temperature. Reformation of a3FeCO after photolysis is an apparent first order process below 210 K with a distribution of rate constants. The kinetics are well described by a power law. Arrhenius behavior is followed between 140 K and 180 K to yield a peak activation enthalpy of 40.3 kJ/mol and a distribution in g(H) = 2.56 kJ/mol (full width at half-maximum). The major component of a3FeCO shows a very narrow CO absorption band (full width at half-maximum = 2.4 cm-1), while that of CuBCO shows a broader CO absorption (full width at half-maximum = 6 cm-1). These data indicate that in the reduced carbon monoxide complex, a3FeCO is in highly ordered nonpolar surroundings sufficiently separated from CuB that it is not perturbed by motion of the latter, while CuBCO is in less ordered, more flexible surroundings. PMID- 6276396 TI - Clonal variation in adrenocorticotropin-induced desensitization of adenylate cyclase in Y1 adrenocortical tumor cells. Association with a 68,000-dalton protein. AB - Adrenocorticotropin(ACTH)-induced desensitization of adenylate cyclase was examined in subclones derived from the ACTH-responsive, Y1 mouse adrenocortical tumor cell line. This report describes clonal variation in ACTH-induced desensitization of adenylate cyclase and an associated variation in the level of a 68,000-dalton protein, p68. A subclone of Y1 cells with a low level of p68 (0.8% of total protein) exhibited a faster rate of desensitization and a slower rate of recovery from desensitization when compared with a clone containing a high level of p68 (10% of total protein). In three clones with low levels of p68, ACTH desensitized adenylate cyclase with ED50 values from 0.3 to 0.5 nM. In several clones with high levels of p68, the adenylate cyclase system was more resistant to ACTH-induced desensitization; the ED50 values for ACTH in these clones ranged from 2 to 12 nM. Among 11 ACTH-responsive subclones, the level of p68 correlated significantly (p less than 0.001, r = 0.87) with resistance to the desensitization induced by 1 nM ACTH. These results suggest that p68 may function in the maintenance of an ACTH-responsive adenylate cyclase system, or that the level of p68 and responsiveness to ACTH are coordinately regulated. PMID- 6276397 TI - Human serum albumin: an allosteric domain model for bilirubin binding specificity. An enantiomeric spin label study. PMID- 6276398 TI - Site-specific cyclic nucleotide binding and dissociation of the holoenzyme of cAMP-dependent protein kinase. AB - The photoaffinity reagent 8-azidoadenosine 3':5'-monophosphate (8-N3cAMP) was previously shown to modify a single tyrosine residue on the type II regulatory subunit of cAMP-dependent protein kinase (Kerlavage, A.R., and Taylor, S.S. (1980) J. Biol. Chem, 255, 8483-8488). In the present studies, the binding stoichiometries of type II holoenzyme for cAMP and 8-N3cAMP were determined using Millipore filtration assays in the absence (Assay A) and presence (Assay B) of 2 M NaCl and histone. The binding stoichiometry of holoenzyme for cAMP was 2 mol/mol with Assay A, and 4 mol/mol with assay B. The binding stoichiometry for 8 N3cAMP was 2 mol/mol with Assay B or with Assay A following photolysis of the holoenzyme:8-N3cAMP mixture. In the absence of photolysis, the binding stoichiometry for 8-N3cAMP was 0.4 mol/mol with Assay A. Both 8-N3cAMP and cAMP fully dissociated the holoenzyme. Holoenzyme, labeled with 8-N3[3H]cAMP on a preparative scale, incorporated 1 mol of 8-N3[3H]cAMP/mol of regulatory subunit (RII) monomer. The labeled RII was separated from catalytic subunit, cleaved with cyanogen bromide, and the resultant peptides were separated by high performance liquid chromatography. A single radioactive peptide was observed which had the same NH2 terminal residue and amino acid composition as the peptide obtained when dissociated RII was labeled with 8-N3cAMP. PMID- 6276399 TI - The activity of 5'-nucleotidase in liver plasma membranes is affected by the increase in bilayer fluidity achieved by anionic drugs but not by cationic drugs. AB - The 5'-nucleotidase activity of rat liver plasma membranes could be selectively modulated by the anionic drugs phenobarbital and pentobarbital, whereas the corresponding activity of a Lubrol-solubilized preparations remained unaltered. The perturbation in the outer half of the bilayer induced by phenobarbital, which lead to a depression in the high temperature onset of the lipid phase separation occurring in this half of the bilayer, concomitantly lowered the break temperature in Arrhenius plots of 5'-nucleotidase activity from 28 degrees C to 16 degrees C. The stimulation of the membrane-bound activity achieved by low anionic drug concentrations was attributed to a preferential fluidization of the outer half of the bilayer. Contrarily, the cationic drugs prilocaine and carbocaine, when tested over agent concentrations that dramatically increase the fluidity of the inner half of the bilayer, achieved no selective effects on the membrane-bound enzyme. Prilocaine (10 mM) was previously found to induce a lipid phase separation at 11 degrees C that was attributed to the lipids of the internal (cytosol-facing) half of the bilayer, but had no effect on the onset of the lipid phase separation occurring at 28 degrees C. Since Arrhenius plots of 5' nucleotidase activity in the presence of 10 mM prilocaine concentrations demonstrated only the single break at 28 degrees C, we suggest that prilocaine is unable to selectively perturb the enzyme because this cationic drug preferentially interacts with the acidic phospholipids residing in the inner half of the bilayer. The activity of the ectoenzyme 5'-nucleotidase in rat liver plasma membranes appears to be regulated by the external half of the bilayer only. These results support the view that independent modulation of he fluidity or chemical constituents of each half of the bilayer can distinctly affect the activity of proteins that are themselves asymmetrically orientated within the bilayer. PMID- 6276400 TI - Calcium binding domains of calmodulin. Sequence of fill as determined with terbium luminescence. AB - Terbium, a trivalent lanthanide, effectively substituted for Ca2+ in calmodulin as judged by several criteria: intrinsic fluorescence spectra, altered mobilities on polyacrylamide gel electrophoresis, formation of a stable complex with troponin I or calcineurin, and stimulation of phosphodiesterase. Calmodulin harbors four Ca2+ binding domains; domains I and II contain no tyrosine, whereas domains III and IV each have one tyrosine. The binding of Tb3+ to calmodulin was followed by the increase of Tb3+ fluorescence at 545 nm upon binding to calmodulin. This fluorescence was elicited either by exciting Tb3+ directly at 222 nm or by exciting the calmodulin tyrosine at 280 nm with resulting energy transfer from tyrosine to Tb3+. Fluorescence generated by direct excitation measures binding of Tb3+ to any of the Ca2+ binding domains, whereas energy transfer through indirect excitation is effective only when Tb3+ is within 5 A of tyrosine, indicating that Tb3+ necessarily occupies a Ca2+ binding domain that contains tyrosine. A judicious use of the direct and indirect excitation could reveal the sequence of fill of the binding domains. Our results suggest these domains are filled in the following sequence: 1) domain I or II; 2) domains III and IV; and 3) domain II or I that has not been filled initially. PMID- 6276401 TI - EPR signal, purple color, and iron binding in porcine uteroferrin. AB - Titration of purple uteroferrin with two reducing equivalents of ferrous ion doubles the intensity of its g' = 1.74 EPR signal while inducing only minor changes in spectral characteristics. Unexpectedly, intensification of the EPR signal is not accompanied by a commensurate increase in visible absorption: only a small shift in peak position from 545 to 525 nm is observed. These observations suggest that uteroferrin can bind a second iron to form a paramagnetic complex that is essentially nonchromophoric. Titration of the pink protein with one oxidizing equivalent of ferric ion also doubles the intensity of its g' = 1.74 EPR signal, again shifting the primary visible absorption band to 525 nm. In either case, therefore, it is possible to augment the g' = 1.74 EPR signal without a corresponding augmentation of purple-pink color. Conversely, the addition of hydrogen peroxide (but not ferricyanide) to purple uteroferrin obliterates the g' = 1.74 EPR signal without abolishing the protein's visible absorption spectrum. Hence, it is also possible to have purple color without a corresponding g' = 1.74 EPR signal. To explain these curious results, two possible models, one involving low spin ferric iron exclusively and the other a combination of low spin ferric and high spin ferrous, are adduced. PMID- 6276402 TI - Nucleotide sequence of the Escherichia coli polA gene and primary structure of DNA polymerase I. AB - We report the nucleotide sequence of 3.2 kilobase pair region of the Escherichia coli polA gene, comprising the coding region for DNA polymerase I with about 400 base pairs of flanking sequence. The amino acid sequence for DNA polymerase I derived from our DNA sequence is largely consistent with previous protein chemical data. In the following paper, Brown et al. (Brown, W. E., Stump, K. H., and Kelley, W. S. (1982) J. Biol. Chem. 257, 1965-1972) present additional protein chemistry experiments that further confirm our sequence. Mild proteolysis of DNA polymerase I is known to produce two enzymatically active fragments (Brutlag, D., Atkinson, M. R., Setlow, P., and Kornberg, A. (1969) Biochem. Biophys. Res. Commun. 37, 982-989; Klenow, H., and Henningsen, I. (1970) Proc. Natl. Acad. Sci. U. S. A. 74, 5632-5636). We have located the site of this cleavage between residues 323 and 324 of the 928 amino acid polymerase molecule. By sequence comparison of the polA1 and wild type alleles, we have identified the polA1 mutation as a change from Trp (TGG) to amber (TAG) at residue 342. PMID- 6276403 TI - Purification and characterization of a cyclic GMP-stimulated cyclic nucleotide phosphodiesterase from bovine tissues. AB - A cGMP-stimulated cyclic nucleotide phosphodiesterase has been purified to near homogeneity from bovine adrenal and heart tissues. The purification procedure utilizes chromatography on DEAE-cellulose and cGMP affinity resin. The procedure can be completed within 2 days and is easily adapted to large scale. To obtain pure enzyme, an 8,000-9,000-fold increase in specific activity was required in adrenal tissues and 15,000-30,000-fold in cardiac muscle. A single band of protein having an apparent Mr = 105,000-107,000 was seen on sodium dodecyl sulfate gel electrophoresis. At equilibrium, native polyacrylamide gradient gel electrophoresis revealed a single major band having an apparent Mr = 240,000. Cyclic GMP binding and phosphodiesterase activity co-migrated with the protein band on native polyacrylamide gradient gels. The enzyme bound cGMP with high affinity reaching a maximum binding of 1.02 mol of cGMP bound/mol of enzyme dimer. Titration curves of the binding data indicated at least two classes of binding sites with 10% maximal binding occurring at 7 nM and 90% maximal binding at 4 microM cGMP. Kinetic analysis indicated the enzyme can hydrolyze both cAMP and cGMP with similar maximal rates. The nucleotide concentration at half-maximal velocity were 30 and 10 microM for cAMP and cGMP, respectively. The hydrolyses of both nucleotides exhibit positive homotropic cooperativity with Hill coefficients of 1.9 for cAMP and 1.3 for cGMP. The rate of cAMP hydrolysis by the purified enzyme when measured at 10 microM cAMP was enhanced 5- to 6-fold by low levels of cGMP. PMID- 6276404 TI - Quantitative resolution of succinate-cytochrome c reductase into succinate ubiquinone and ubiquinol-cytochrome c reductases. AB - A purified, active succinate-ubiquinone reductase was prepared from succinate cytochrome c reductase without damage to ubiquinol-cytochrome c reductase by 1.1% Triton X-100 solubilization at pH 8.0, and calcium phosphate column chromatography in 50 mM Tris-succinate buffer, pH 8.0, containing 30 mM potassium phosphate. Succinate-ubiquinone reductase thus obtained contains ubiquinone and catalyzes thenoyltrifluoroacetone-sensitive oxidation of succinate by 2,6 dichlorophenolindophenol in the absence of exogenous mediator. Addition of ubiquinone enhanced the activity about 50%. Analytical sodium dodecyl sulfate polyacrylamide gel electrophoresis showed that the enzyme contains four polypeptides. The high molecular weight polypeptide contaminants usually observed in the Complex II preparation obtained by the reported method were absent. The active succinate-ubiquinone reductase can reconstitute with the cytochrome b c1III complex, or Complex III to form succinate-cytochrome c reductase in the absence of exogenous ubiquinone or with the resolved ubiquinol-cytochrome c reductase in the presence of ubiquinone and phospholipids. Under the proper conditions, all the original succinate-cytochrome c reductase was obtained, indicating that the resolution caused no damage to the protein, despite the removal of phospholipids and ubiquinone from the ubiquinol-cytochrome c reductase region. PMID- 6276405 TI - N6,O2'-dibutyryl cycle AMP and glucose regulate the amount of messenger RNA coding for hepatic phosphoenolpyruvate carboxykinase (GTP). AB - Rat liver phosphoenolpyruvate carboxykinase (GTP) (EC 4.1.1.32) mRNA was purified to 25% of total mRNA activity (greater than 50-fold enrichment) by polysome immunoprecipitation. This preparation was used as template for the synthesis of cDNA that was subsequently cloned in Escherichia coli. The resulting clones were screened by in situ hybridization and by hybrid-selected translation of phosphoenolpyruvate carboxykinase mRNA. The cDNA insert of one plasmid, pPC2, was complementary to phosphoenolpyruvate carboxykinase mRNA as determined by these screening procedures. pPC2 cDNA was 760 base pairs in length and a partial restriction enzyme map was constructed. pPC2 was labeled with 32P by nick translation and was used as a hybridization probe to quantitate phosphoenolpyruvate carboxykinase mRNA following N6,O2'-dibutyryl cAMP (Bt2cAMP) injection or glucose feeding. Bt2cAMP increased whereas glucose decreased the level of hybridizable phosphoenolpyruvate carboxykinase mRNA and in all cases the changes were proportional to the in vitro translational activities measured in a reticulocyte lysate system. The half-life of phosphoenolpyruvate carboxykinase mRNA sequences was measured by an indirect procedure involving their quantitation, by hybridization assay, during deinduction and induction. The half life was approximately 10-40 min during deinduction by glucose or during induction stimulated by Bt2cAMP. Our data indicate that cAMP enhances some step in the generation of phosphoenolpyruvate carboxykinase mRNA. PMID- 6276406 TI - Selection of the nuclear gene for the mitochondrial adenine nucleotide translocator by genetic complementation of the op1 mutation in yeast. AB - Oxidative phosphorylation 1 (OP1), a nuclear gene of yeast Saccharomyces cerevisiae which is required for the expression of a functional mitochondria, has been isolated on a recombinant plasmid. The gene was selected from a recombinant plasmid pool which contained wild type yeast genomic DNA by transformation of the yeast nuclear mutant (op1) followed by a two-stage screening procedure. A recombinant plasmid containing a 2.6 kilobase Bam HI fragment of genomic yeast DNA inserted in either orientation into the single Bam HI site of yeast vector YEp 13 could complement the op1 mutation. Analysis of the gene product of this inserted DNA by three independent methods, 1) in vivo expression in Escherichia coli maxicells, 2) cell-free translation of plasmid selected RNA, and 3) expression analysis in yeast, revealed that its gene product is a protein of Mr = 30,000-32,000, which cross-reacts with specific anti-serum to the adenine nucleotide translocator of the mitochondrial inner membrane. The selection procedure is efficient and can be used for the isolation of any defined yeast nuclear gene which participates in mitochondrial development. PMID- 6276408 TI - Granular-cell tumors of the esophagus and common bile duct. PMID- 6276407 TI - Effects of cholinergic and adrenergic agonists on phosphorylation of a 165,000 dalton myofibrillar protein in intact cardiac muscle. AB - The purpose of this investigation was to examine the effects of beta-adrenergic and muscarinic cholinergic agonists on protein phosphorylation in intact frog atrium. beta-Adrenergic agonists increase and muscarinic agonists decrease 32P incorporation into a 165,000-dalton (165K) protein within less than 1 min. The concentrations of isoproterenol that produce increases in 32P incorporation into the 165K protein and in systolic tension are similar. Further, the changes in 32P incorporation and tension produced by isoproterenol occur with similar time courses. Carbamylcholine decreases tension somewhat more quickly and at lower concentrations than it decreases 32P incorporation, however. Isoproterenol stimulated 32P incorporation is thought to be mediated by cAMP-dependent protein kinase because bath application of dibutyryl cAMP, cholera toxin, or phosphodiesterase inhibitors increase 32P incorporation into the 165K protein in intact atria. When heart homogenates are incubated in the presence of [gamma 32P]ATP, cAMP stimulates the incorporation of 32P into the 165K protein. cGMP is much less effective. We suggest that carbamylcholine decreases 32P incorporation into the 165K protein by a mechanism independent of cAMP levels because carbamylcholine inhibits the stimulation of 32P incorporation into the 165K band produced by 8-bromo cAMP in intact cells. Phosphorylation of the 165K protein occurs in cardiac muscle but not in other tissues. We hypothesize that the 165K protein is C-protein, because the 165K- and C-proteins have similar solubilities and are associated with the myofibril. Further, antibodies produced against the 165K protein bind to C-protein purified from rabbit heart and also bind to the same region of the myofibril where C-protein is found. PMID- 6276409 TI - Primary role of sarcoplasmic reticulum in phasic contractile activation of cardiac myocytes with shunted myolemma. AB - Homogeneous populations of single myocytes showing good preservation of ultrastructure were obtained by enzymatic digestion of rabbit and rat hearts, and maintained in a relaxed state in the presence of free Ca2+ concentrations less than 10(-7) M. Ultrastructural details such as a cytoskeleton of 100-A filaments connected to the sarcolemma at the Z lines were demonstrated especially well in these preparations. In spite of seemingly normal structure, electron probe analysis of cryosections reveals similar concentrations of electrolytes in the medium and in the cytoplasm, indicating the presence of electrochemical shunting across the external membrane. The dissociated myocytes display Ca uptake and phasic contractions that are apparently dependent on mitochondrial respiration, but are not affected by mitochondrial uncouplers when ATP and phosphocreatine are added. The uptake is augmented by oxalate and, based on identification of calcium oxalate crystals by electron microscopy and electron probe analysis, is localized to the sarcoplasmic reticulum (SR). An advantageous feature of the dissociated myocytes is that they are suitable for experiments using large numbers of cells in suspension. Thereby, velocities of calcium transport were measured directly by isotopic tracer and filtration methods. It was then found that the lowest CA2+ concentrations (5 x 10(-7) M for the rabbit and 1 x 10(-7) M for the rat) sustaining Ca transport also induce phasic contractile activity in all myocytes, even though the external membrane is electrochemically shunted. A stepwise rise in the Ca2+ concentration of up to one order of magnitude, increases transport velocities in parallel with the rates of phasic contractions. Both these parameters are affected by Mg2+, temperature, cyclic-AMP, and methylxanthines, even though the Ca2+ concentration is maintained constant in the medium. Therefore, Ca transport by SR is a requirement and a rate limiting factor for the occurrence of phasic contractile activation in dissociated cardiac cells retaining an electrochemically shunted external membrane. It is suggested that transient Ca release required for phasic contractile activation is due to equilibrium oscillations across the SR membrane. The sequential pattern of sarcomere activation is consistent with a self propagating mechanism of calcium release. SR in dissociated skeletal muscle cells sustains a greater Ca transport activity than in dissociated heart cells. However, the heart cells display a much higher phasic contractile activity, indicating that cardiac SR has a greater tendency to release accumulated calcium. If free Ca2+ in the medium is raised above 10(-6) M, both cardiac and skeletal myocytes undergo contractures and degenerative phenomena, accompanied by Ca, Mg, and phosphate accumulation in cardiac mitochondria. PMID- 6276410 TI - Ultrastructural localization of Na+,K+-ATPase in rat and rabbit kidney medulla. AB - Na+,K+-ATPase was localized at the ultrastructural level in rat and rabbit kidney medulla. The cytochemical method for the K+-dependent phosphatase component of the enzyme, using p-nitrophenylphosphate (NPP) as substrate, was employed to demonstrate the distribution of Na+, K+-ATPase in tissue-chopped sections from kidneys perfusion-fixed with 1% paraformaldehyde-0.25% glutaraldehyde. In other outer medulla of rat kidney, ascending thick limbs (MATL) were sites of intense K+-dependent NPPase (K+-NPPase) activity, whereas descending thick limbs and collecting tubules were barely reactive. Although descending thin limbs (DTL) of short loop nephrons were unstained, DTL from long loop nephrons in outer medulla were sites of moderate K+-NPPase activity. In rat inner medulla, DTL and ascending thin limbs (ATL) were unreactive for K+-NPPase. In rabbit medulla, only MATL were sites of significant K+-NPPase activity. The specificity of the cytochemical localization of Na+,K+-ATPase at reactive sites in rat and rabbit kidney medulla was demonstrated by K+-dependence of reaction product deposition, localization of reaction product (precipitated phosphate hydrolyzed from NPP) to the cytoplasmic side of basolateral plasma membranes, insensitivity of the reaction to inhibitors of nonspecific alkaline phosphatase, and, in the glycoside sensitive rabbit kidney, substantial inhibition of staining by ouabain. The observed pattern of distribution of the sodium transport enzyme in kidney medulla is particularly relevant to current models for urine concentration. The presence of substantial Na+,K+-ATPase in MATL is consistent with the putative role of this segment as the driving force for the countercurrent multiplication system in the outer medulla. The absence of significant activity in inner medullary ATL and DTL, however, implies that interstitial solute accumulation in this region probably occurs by passive processes. The localization of significant Na+,K+ ATPase in outer medullary DTL of long loop nephrons in the rat suggests that solute addition in this segment may occur in part by an active salt secretory mechanism that could ultimately contribute to the generation of inner medullary interstitial hypertonicity and urine concentration. PMID- 6276411 TI - Distribution of cells bearing receptors for a colony-stimulating factor (CSF-1) in murine tissues. AB - CSF-1 is a subclass of the colony-stimulating factors that specifically stimulates the growth of mononuclear phagocytes. We used the binding of 125I-CSF 1 at 0 degrees C by single cell suspensions from various murine tissues, in conjunction with radioautography, to determine the frequency of binding cells, their identity, and the number of binding sites per binding cell. For all tissues examined, saturation of binding sites was achieved within 2 h at 2--3 x 10(-10) M 125I-CSF-1. The binding was irreversible and almost completely blocked by a 2 h preincubation with 5 x 10(-10) M CSF-1. 125I-CSF-1 binding was exhibited by 4.3% of bone marrow cells, 7.5% of blood mononuclear cells, 2.4% of spleen cells, 20.5% of peritoneal cells, 11.8% of pulmonary alveolar cells and 0.4% of lymph node cells. Four morphologically distinguishable cell types bound 125I-CSF-1: blast cells; mononuclear cells with a ratio of nuclear to cytoplasmic area (N/C) greater than 1; cells with indented nuclei; and mononuclear cells with N/C less than or equal to 1. No CSF-1 binding cells were detected among blood granulocytes or thymus cells. Bone marrow promyelocytes, myelocytes, neutrophilic granulocytes, eosinophilic granulocytes, nucleated erythroid cells, enucleated erythrocytes, and megakaryocytes also failed to bind. The frequency distribution of grain counts per cell for blood mononuclear cells was homogenous. In contrast, those for bone marrow, spleen, alveolar, and peritoneal cells were heterogeneous. The monocytes in blood or bone marrow (small cells, with either indented nuclei or with N/C greater than 1) were relatively uniformly labeled, possessing approximately 3,000 binding sites per cell. Larger binding cells (e.g., alveolar cells) may possess higher numbers of receptors. It is concluded that CSF-1 binding is restricted to mononuclear phagocytic cells and their precursors and that it can be used to identify both mature and immature cells of this series. PMID- 6276412 TI - Calmodulin is a major component of extruded trichocysts from Paramecium tetraurelia. AB - Extruded trichocysts are composed of a family of proteins with molecular weights between 15,000 and 20,000. We have used heat treatment and affinity chromatography on fluphenazine-Sepharose to purify calmodulinlike proteins from whole cells and from extruded trichocysts. The purified protein from trichocysts is indistinguishable from that of whole cells; it is heat-stable, activates brain phosphodiesterase in a Ca++-dependent fashion, changes mobility on SDS polyacrylamide gels in the presence of Ca++, contains 1 mol of trimethyllysine/17 kdaltons, and has the amino acid composition characteristic of calmodulins. Calmodulin is a major component of purified, extruded trichocysts, of which it represents between 1 and 10% by mass. The other proteins of the trichocyst also resemble calmodulin in several properties. Possible roles for calmodulin in the Ca++-activated extrusion of trichocysts is discussed. PMID- 6276413 TI - Photosensitive phosphoproteins in Halobacteria: regulatory coupling of transmembrane proton flux and protein dephosphorylation. AB - A photoregulated reversible protein phosphorylation system controlled by the halobacterial rhodopsins was recently reported. The results presented in this paper identify the initial steps in the pathway from the absorption of light to the photoregulated protein phosphorylation and dephosphorylation reactions. Action spectrum, biochemical, and genetic analyses show that the proton pump bacteriorhodopsin mediates light-induced dephosphorylation of three photoregulated phosphoproteins. Light absorbed by bacteriorhodopsin is used to establish a proton efflux from the cells. The increase in the inwardly directed protonmotive force (pmf) from this efflux induces dephosphorylation of the three phosphoproteins, as demonstrated by the effects of the protonophore CCCP and of artificially imposed transmembrane pH gradients. Upon darkening the cells, cessation of the proton efflux through bacteriorhodopsin causes a decrease in pmf, which induces rephosphorylation of the proteins. Pmf appears to function as a regulator rather than a driving force in this system. Measurements of pmf driven ATP synthesis in our conditions indicate the regulation of protein phosphorylation by pmf is probably not a consequence of proton flux through the H+ ATPase, a known energy coupling structure in these cells. The properties of this system may indicate the existence of a pmf detector which regulates kinase or phosphatase activity; i.e., a regulatory coupling device. PMID- 6276414 TI - Identification of gelsolin, a Ca2+-dependent regulatory protein of actin gel-sol transformation, and its intracellular distribution in a variety of cells and tissues. AB - Antiserum prepared against gelsolin, a major Ca2+-dependent regulatory protein of actin gel-sol transformation in rabbit lung macrophages, was used to detect the presence of proteins immunologically related to gelsolin in a variety of cells and tissues. Cell extracts were electrophoresed on polyacrylamide gels, and replicas of the gels on cellulose nitrate paper were stained by an indirect immunohistochemical technique. A single band of crossreactive material which comigrates with macrophage gelsolin is found in at least nine different kinds of cells and tissues derived from rabbits and humans and in four lines of cultured cells from humans and rats. Gelsolin was also identified in human serum and plasma, raising the possibility that it may contribute to the clearance of actin from the circulatory system. Using this antiserum, we demonstrated, by indirect immunofluorescent staining of acetone-fixed macrophages and polymorphonuclear leukocytes, that gelsolin resides in the cortical cytoplasm and that during phagocytosis it is concentrated in pseudopodia engulfing particles to be ingested, an area of the cytoplasm actively engaged in movement. In longitudinal cryostat sections of contracted rabbit skeletal muscle, antigelsolin staining was associated with the I-band of the myofibril, suggesting that it may be involved, by an as yet undefined mechanism, in skeletal muscle function. In rabbit intestinal epithelial cells, gelsolin was associated with the cytoplasm and the terminal web region of the brush border, a localization distinct from that previously reported for villin, a structurally and functionally similar protein isolated from the brush borders of chicken intestinal epithelial cells. In conclusion, our findings support the idea that gelsolin is involved in the regulation of movement and suggest that gelsolin-mediated Ca2+-regulation of actin cytoskeletal structure, first characterized in macrophages, may be of general importance. PMID- 6276416 TI - Properties of a sarcoma-growth-factor-like peptide from cells transformed by a temperature-sensitive sarcoma virus. AB - Serum-free conditioned media was collected from three sarcoma virus-transformed cell lines and an untransformed cell line. All three virally transformed lines produced and released growth factors into their serum-free media. The major activity in all cases, whether the cells were transformed by Moloney sarcoma virus (MSV) or Kirsten sarcoma virus (KiSV), or whether they were mouse or rat, was a sarcoma-growth-factor (SGF)-like activity with an apparent molecular weight of 10,000. The SGF-like pools from a Moloney sarcoma virus-transformed mouse 3T3 cell and a Kirsten sarcoma virus-transformed NRK cell were further purified by carboxymethyl cellulose chromatography. The elution profiles of these peptides were very similar. The serum-free conditioned media from the untransformed cells showed no detectable growth stimulating activity. The temperature sensitivity of an SGF-like growth factor from the supernate of a NRK cell transformed by a wild type Kirsten sarcoma virus (KiSV) was compared with that of the SGF-like activity from the supernates of a NRK cell transformed by a ts-mutant of KiSV that is temperature sensitive with respect to transformation (ts-371 Cl 5). Neither the cells transformed by the wild-type sarcoma virus nor those transformed by the temperature sensitive virus released a SGF-like activity that was temperature sensitive under the conditions of the assays. PMID- 6276415 TI - Kinetic analysis of chemotactic peptide receptor modulation. AB - The dynamics of the chemotactic peptide receptor on rabbit peritoneal polymorphonuclear leucocytes were followed using the tritiated peptide N formylnorleucylleucylphenylalanine (FNLLP). We have used a kinetic analysis to examine the possible interrelationships between receptor loss (down-regulation), receptor-mediated peptide uptake, and receptor recycling. We have previously demonstrated that cells incubated with FNLLP show a dose-dependent reduction in the number of receptors available on the surface. This receptor down-regulation is complete within 20 min and then the number of receptors available for binding remains at a plateau level. Peptide continues to be taken up in a receptor mediated manner even after down-regulation is complete. If peptide is removed, receptor recovery occurs and does not require protein synthesis. In these studies we have investigated the kinetics of these processes. On the basis of this analysis, we propose that the plateau receptor level is a steady-state in which receptor internalization and return occur continuously. We demonstrate that the rate of receptor-mediated peptide uptake is approximately equal to the rate of receptor recovery measured after peptide removal. In addition, the rate of receptor recovery is proportional to the number of receptors missing from the surface, suggesting receptor recycling may be occurring. PMID- 6276417 TI - Patterns of cell communication and differentiation in SV40 transformed human keratinocytes. AB - Fluorescein dye microinjection was used to demonstrate changes in communication between human epidermal keratinocytes grown in vitro after infection by the oncogenic virus, SV40. Whereas keratinocytes are normally fully coupled to each other, dye spread becomes progressively restricted to small cell subpopulations after infection, although dye coupling is increased when the infected cells attain high densities. Reduction or enhancement of dye coupling is correlated with similar changes in the extent of cytochemical differentiation and cornified envelope formation. PMID- 6276418 TI - Clonal variation in cultured neuroblastoma cells. II. The relationship of increased intracellular cyclic AMP content to increased anchorage requirement for growth and flattened morphology. AB - Flat variant clones were isolated from both the N18 neuroblastoma cell line and from a subclone of the parent line exhibiting the typical round cell morphology. Several revertant clones exhibiting the round parental morphology were also isolated from one of the flat variant clones. The flat variants exhibit decreased cloning efficiency in suspension and increased amounts of myosin heavy chain when compared to the round cell clones. Intracellular cAMP levels were increased from two- to fivefold over those in clones representative of the parent line and in the round cell revertants. Treatment with the phosphodiesterase inhibitor RO20 1724 increased cAMP levels and reduced suspension cloning efficiency without altering doubling time or attached cloning efficiency. Increasing cAMP levels of two of the round cell clones by treatment with the phosphodiesterase inhibitor caused increased flattening of the cell body and increased myosin heavy chain content. Thus, even though increased cAMP level may be sufficient to explain the reduced cloning efficiency of the flat variants, it is not the sole cause of the flat morphology. PMID- 6276419 TI - Cytochalasin inhibition of isolated rat gastric parietal cell function. AB - Submicrogram concentrations (0.04-0.29 microM) of the microfilament disrupting agents cytochalasins D, E, and B (CD, CE, CB) were shown to inhibit secretagogue stimulated 14C-aminopyrine accumulation (AP) in isolated rat gastric mucosal parietal cells. The microtubule disrupting agent colchicine had little influence on AP accumulation. Histamine- and dibutyryl cyclic AMP (DbcAMP)-stimulated AP accumulation was inhibited with an order of potency CD greater than CE approximately equal to CB. CB inhibition of these secretagogue actions was, however, only approximately 65-70% of the maximal stimulated response, whereas CD and CE caused 100% inhibition. On the other hand, carbamylcholine-stimulated AP accumulation was inhibited 100% by all cytochalasins tested with an order of potency CD approximately equal to CE greater than CB. These data are discussed in relation to acid secretagogue-induced morphological changes involving actin filament organization in parietal cells. PMID- 6276420 TI - Cell damage in the brain: a speculative synthesis. PMID- 6276421 TI - Autonomic nerves in the mammalian choroid plexus and their influence on the formation of cerebrospinal fluid. AB - The choroid plexuses of all ventricles receive a well-developed adrenergic and cholinergic innervation reaching both the secretory epithelium and the vascular smooth muscle cells. Also peptidergic nerves, containing vasoactive intestinal polypeptide, are present but primarily associated only with the vascular bed. A sympathetic inhibitory effect on the plexus epithelium has been indicated in determinations of carbonic anhydrase activity and by studies of various aspects of active transport in isolated plexus tissue. Pharmacological analysis in vitro has shown the choroidal arteries to possess both vasoconstrictory alpha adrenergic and vasodilatory beta-adrenergic receptors. Electrical stimulation of the sympathetic nerves, which originate in the superior cervical ganglia, induces as much as 30% reduction in the net rate of cerebrospinal fluid (CSF) production, while sympathectomy results in a pronounced increase, about 30% above control, in the CSF formation. There is strong reason to believe that the choroid plexus is under the influence of a considerable sympathetic inhibitory tone under steady state conditions. From pharmacological and biochemical experiments it is suggested that the sympathomimetic reduction in the rate of CSF formation is the result of a combined beta-receptor-mediated inhibition of the secretion from the plexus epithelium and a reduced blood flow in the choroid plexus tissue resulting from stimulation of the vascular alpha-receptors. The choroid plexus probably also represents an important inactivation site and gate mechanism for sympathomimetic amines, as evidenced by considerable local activity of catechol-O methyl transferase and monoamine oxidase, primarily type B. The CSF production rate is also reduced by cholinomimetic agents, suggesting the presence of muscarinic-type cholinergic receptors in the choroid plexus. PMID- 6276422 TI - Hypoglycemic brain injury: phospholipids, free fatty acids, and cyclic nucleotides in the cerebellum of the rat after 30 and 60 minutes of severe insulin-induced hypoglycemia. PMID- 6276423 TI - A fast and simple high-performance liquid chromatographic assay for aryl hydrocarbon hydroxylase. AB - A simple high-performance liquid chromatographic method using fluorescence detection of the remaining substrate is described for the determination of benzo[alpha]pyrene hydroxylase activity. This assay is far simpler than the previous ones, as it does not require extraction or centrifugation and the measurement occurs directly after dilution of the total incubation medium. The aryl hydrocarbon hydroxylase (AHH) activities in rat liver microsomes are in agreement with those obtained by radioactive assays. Moreover, this assay allows the routine determination of the AHH activity in animal tissues. PMID- 6276424 TI - Determination of amdinocillin in plasma and urine by high-performance liquid chromatography. AB - A rapid, sensitive and specific high-performance liquid chromatographic (HPLC) assay was developed for the determination of amdinocillin (formerly mecillinam) in human plasma and urine. The assay is performed by direct injection of a plasma protein-free supernatant or a dilution of urine. A 10 micrometer muBondapak phenyl column with an eluting solvent of water--methanol--1 M phosphate buffer, pH 7 (70:30:0.5) was used, with UV detection of the effluent at 220 nm. Azidocillin potassium salt [potassium-6-(D-(-)-alpha-azidophenyacetamido) penicillanate] was used as the internal standard and quantitation was based on peak height ratio of amdinocillin to that of the internal standard. The assay has a recovery of 74.4 +/- 6.3% (S.D.) in the concentration ranges of 0.1-20 microgram per 0.2 ml of plasma with a limit of detection equivalent to 0.5 microgram/ml plasma. The urine assay was validated over a concentration range of 0.025-5 mg/ml of urine, and has a limit of detection of 0.025 mg/ml (25 microgram/ml) using a 0.1-ml urine specimen per assay. The assay was applied to the determination of plasma and urine concentrations of amdinocillin following intravenous administration of a 10 mg/kg dose of amdinocillin to two human subjects. The HPLC and microbiological assays were shown to correlate well for these samples. PMID- 6276426 TI - Assay for conjugated estrogens in tablets using fused-silica capillary gas chromatography. PMID- 6276425 TI - Marijuana metabolites in urine of man. X. Identification of marijuana use by detection of delta 9-tetrahydrocannabinol-11-oic acid using thin-layer chromatography. AB - Marijuana use can be determined by detecting delta 9-tetrahydrocannabinol-11-oic acid (THC-11-oic acid) in urine. For this, we describe a procedure for its chemical detection by using sequential thin-layer chromatography on a single plate for rapid isolation and identification. A volume of urine containing 50 mg of creatinine is concentrated by evaporation to 10 ml, the concentrate is enzymically hydrolyzed for 30 minutes and extracted with ether, and the extract is purified by treatment with NaHCO3, then chromatographed in an alkaline and an acidic solvent sequence. The plate is sprayed with Fast Blue Salt B, and THC-11 oic is identified by its characteristic mobility and its characteristic colour reaction. The sensitivity is 0.5 microgram. THC-11-oic acid has been detected in urines collected after the smoking of one standard cigarette containing 16-18 mg of delta 9-tetrahydrocannabinol and in 34 of the first 100 tests of spontaneously collected urines of patients in a hospital drug-abuse treatment program. Multiple samples are easily carried through this extraction procedure. Evaporative concentration takes about 20 min per sample, and the analysis of eight concentrated samples take about 5.5 h. PMID- 6276427 TI - Extraction of glucuronide metabolites of delta 9-tetrahydrocannabinol by diethyl ether. PMID- 6276428 TI - A portable device for the rapid concentration of viruses from large volumes of natural freshwater. AB - A portable device for the rapid concentration of viruses from natural freshwaters described and its performance in field use is evaluated. The system handled up to 500 litres of water in less than 90 min at a cost of only 2 pounds per sample. Where the samples contained sufficient bacteriophages for detection by direct plating the apparent phage recoveries were greater than 75%. Plant and animal viruses were also concentrated from waters with this system. PMID- 6276429 TI - A syndrome of periodic adrenocorticotropin and vasopressin discharge. AB - An 8-yr-old girl is presented who had periodic attacks of vomiting, psychotic depression, drowsiness, and hypertension (160/110 mm Hg) for a period of 16 months after head injury. At the initiation of the attack, serum ACTH and vasopressin levels were prominently increased (610 pg/ml and 41 microunits/ml, respectively), followed by hypercortisolemia, hyponatremia, and hypoosmolality in plasma. Serum PRL also was elevated (91 ng/ml). Responses of GH and cortisol to insulin-induced hypoglycemia and those of TSH to TRH were reduced. Urinary excretion of epinephrine and norepinephrine were increased, while dopamine (DA) excretion was reciprocally decreased, resulting in a marked elevation of the epinephrine plus norepinephrine to DA ratio during the episodes (0.4-4.5); this was normalized on attack-free days (0.08-0.25). During the attack, the concentration of homovanillic acid, a major metabolite of DA in the brain, also was reduced in cerebrospinal fluids from 70 to 23 ng/ml. The administration of methyl-dopa and reserpine effectively suppressed the recurrence of the episode. Although the exact cause of this syndrome is unknown, a periodic metabolic dysfunction of catecholamine in the central nervous system might be postulated. PMID- 6276430 TI - gamma-Aminobutyric acid- and benzodiazepine-binding sites in human anterior pituitary tissue. AB - The existence of a gamma-aminobutyric acid (GABA) system in human anterior pituitary tissue was examined. Crude membrane fractions prepared from human anterior pituitary tissue bound tritiated GABA. The binding was saturable, and Scatchard analysis indicated a single binding site of high affinity (Kd = 40 nM) and a maximum binding of 1.2 pmol/mg protein. Binding was displaced in a dose related manner by the GABA agonists muscimol (KI = 1 X 10(-8) M), isoguvacine (KI = 6 X 10(-7) M), THIP (4,5,6,7-tetrahydroisoxazolo-[5,4-c]pyridin-3-ol); KI = 5 X 10(-6) M), and the antagonist (+)bicuculline (KI = 5 X 10(-5) M) but not its inactive stereoisomer (-)bicuculline (KI greater than 10(-3) M). In anterior pituitary tissue, a significant concentration of GABA was found (mean, 2.5 +/- 0.5 nmol/mg protein) but no glutamic acid decarboxylase activity, the enzyme synthesizing GABA, was detected using a highly sensitive assay. In addition, benzodiazepine binding was present. An affinity of approximately 15 nM and a Bmax of approximately 0.75 pmol/mg protein were observed when using [3H]diazepam as the ligand. No saturable clonazepam binding occurred, and only slight GABA stimulation of diazepam binding was observed (mean, 18%; range, 6-38%). The ability of GABA and benzodiazepine to alter PRL secretion in rats suggests that the human pituitary GABA-binding sites described here might also mediate effects on PRL release. PMID- 6276431 TI - Phosphodiesterase activity in human subcutaneous adipose tissue in hyper- and hypothyroidism. AB - Phosphodiesterase (PDE) activity was determined in sc adipose tissue from 12 patients with untreated hyperthyroidism, 8 patients with untreated hypothyroidism, and 10 healthy nonobese subjects. Eight of the hyperthyroid group were reexamined after treatment when they were euthyroid. The apparent Vmax of the low Km form of PDE was 25% lower in untreated hyperthyroidism (P less than 0.05) and 100% higher in untreated hypothyroidism (P less than 0.01) than in the controls; treatment of the hyperthyroidism resulted in normalization of the value. There was a positive correlation between the mean fat cell volume and the Vmax of the low Km PDE in all conditions (r = 0.64-0.85). As regards the high Km form of PDE, the Vmax values for both hyper- and hypothyroid patients did not differ from those for the control group. The results of this study suggest that in man, the apparent Vmax for the PDE with low Km is determined by the thyroid hormones. PMID- 6276432 TI - Parathyroid hormone effects on serum 1,25-dihydroxyvitamin D levels in patients with X-linked hypophosphatemic rickets: evidence for abnormal 25-hydroxyvitamin D 1-hydroxylase activity. AB - Patients with X-linked hypophosphatemic rickets (XLH) have normal or marginally low serum 1,25-dihydroxyvitamin D [1,25-(OH)2D] levels despite manifesting hypophosphatemia and phosphate depletion, which increase 1,25-(OH)2D production in many animal species. These data are consistent with the possibility that regulation of vitamin D metabolism is abnormal in XLH. However, controversy concerning the role of phosphate in the regulation of 25-hydroxyvitamin D-1 hydroxylase activity in man has raised doubt about this proposed defect. The presence of a defect in vitamin D metabolism could be established if hormonal or metabolic factors, other than hypophosphatemia, were unable to stimulate 25 hydroxyvitamin D-1-hydroxylase activity normally in patients with XLH. Thus, we compared the effects of parathyroid hormone infusion on serum 1,25-(OH)2D levels in patients with XLH and normals. In response to iv infusion of parathyroid extract (200 U at 0915 and 1700 h), the serum 1,25-(OH)2D concentration increased 218% above base line (from 34.0 +/- 3.0 to 108.8 +/- 2.5 pg/ml) in normals and only 68% (from 30.6 +/- 3.0 to 48.8 +/- 5.5 pg/ml) in patients with XLH. The disparate response occurred in spite of an equivalent increase in urinary cAMP excretion in the normals (from 3.00 +/- 0.14 to 8.70 +/- 0.25 mumol/g creatinine . 24 h) and XLH patients (from 3.10 +/- 0.39 to 8.30 +/- 1.0 mumol/g creatinine . 24 h) as well as equivalent decreases in the renal tubular maximum for the reabsorption of phosphate per liter glomerular filtrate (1.2 +/- 0.1 and 0.9 +/- 0.2 mg/dl, respectively). These observations support the possibility that regulation of vitamin D metabolism is abnormal in XLH. PMID- 6276433 TI - Plasma guanosine 3',5'-monophosphate responses to methacholine and epinephrine in humans. AB - The im injection of methacholine into healthy volunteers caused a dose-dependent increase in plasma cGMP levels; this increase was antagonized by atropine, while it was not affected by phentolamine or propranolol. The im injection of epinephrine caused a small but significant rise in plasma cGMP concentrations; this rise was completely blocked by the simultaneous injection of phentolamine, while it was not affected by atropine or propranolol. These data show that changes in the plasma concentration of cGMP may reflect not only cholinergic but also alpha-adrenergic functions in humans. PMID- 6276434 TI - Failure of bromocriptine to lower plasma catecholamines in normal men and women. AB - A plasma catecholamine-lowering effect of bromocriptine has previously been reported in normals, but not in patients with PRL- or GH-secreting pituitary tumors, and has been used as evidence to support the concept of disordered central nervous system dopaminergic tone in patients with such tumors and as a test to distinguish them from normals. In the present study of 16 normal subjects (9 women and 7 men), we found no significant change in plasma norepinephrine or epinephrine after bromocriptine. Similarly, no change occurred in plasma catecholamines in 8 patients with PRL-secreting tumors or 6 patients with ACTH hypersecretion. Our data, therefore, do not provide confirmatory evidence for an effect of bromocriptine on plasma catecholamines in normal subjects and do not support the proposed use of bromocriptine as a test for defective central dopaminergic regulation. PMID- 6276435 TI - [Adenosine 3', 5'-monophosphate (cyclic-AMP) in a persistently infected Vero cells line with measles virus (author's transl)]. PMID- 6276436 TI - [Studies on uremic neuropathy: with reference to uremic toxin (author's transl)]. PMID- 6276437 TI - Selective localization of wild type and mutant mouse hepatitis virus (JHM strain) antigens in CNS tissue by fluorescence, light and electron microscopy. AB - Demyelination may be induced by several different pathogenetic mechanisms. We have been utilizing mouse hepatitis virus (MHV) to study virus-induced demyelination in the central nervous system (CNS). To learn whether the different disease phenotypes in 4-week-old mice, caused by wild type (a model for fatal encephalomyelitis) or mutant ts8 (a model for primary demyelination), is due to an altered cellular tropism, we have developed an immunolabeling technique to evaluate critically the localization of MHV antigens in the unique cells of the CNS. Using mouse derived L-cells and primary neuronal cells in vitro, we determined an appropriate fixative (4% paraformaldehyde and 0.5% glutaraldehyde) that both preserved MHV antigenicity and cell structure. These studies in vitro showed the presence of MHV antigens on the surface of cells. Utilizing immunoperoxidase labeling as developed, we studied the localization of MHV antigens in vivo. MHV antigens associated with wild type (wt) virus were localized in neuronal cells as well as oligodendrocytes, which might account for the encephalomyelitis and primary demyelination, respectively. In contrast, MHV antigens associated with ts8 were localized rarely in neurons but commonly in oligodendrocytes. This might account for the uncommon occurrence of fatal encephalomyelitis, but the frequent presence of primary demyelination. Of interest was the finding of viral antigens during MHV infection in the cytoplasmic processes of oligodendrocytes surrounding intact myelin sheaths. We conclude that the different disease phenotypes caused by wt and mutant ts8 reflect differences in the cellular tropism of the two viruses for cells in the CNS. PMID- 6276439 TI - Retinoic acid. Inhibition of the clonal growth of human myeloid leukemia cells. AB - Vitamin A and its analogues (retinoids) affect normal and malignant hematopoietic cells. We examined the effect of retinoids on the clonal growth in vitro of myeloid leukemia cells. Retinoic acid inhibited the clonal growth of the KG-1, acute myeloblastic leukemia, and the HL-60, acute promyelocytic leukemia, human cell lines. The KG-1 cells were extremely sensitive to retinoic acid, with 50% of the colonies inhibited by 2.4-nM concentrations of the drug. A 50% growth inhibition of HL-60 was achieved by 25 nM retinoic acid. Complete inhibition of growth of both leukemia cell lines was seen with 1 microM retinoic acid. Exposure of KG-1 cells to retinoic acid for only 3-5 d was sufficient to inhibit all clonal growth. The all-trans and 13-cis forms of retinoic acid were equally effective in inhibiting proliferation. Retinal, retinyl acetate, and retinol (vitamin A) were less potent inhibitors. Clonal growth of the human K562 and mouse M-1 myeloid leukemic cell lines was not affected by 10 microM retinoic acid. Retinoic acid also inhibited the clonal growth of leukemia cells from five of seven patients with acute myeloid leukemia. Retinoic acid at concentrations of 5 nM to 0.3 microM inhibited 50% clonal growth, and 1 microM retinoic acid inhibited 64-98% of the leukemic colonies. The inhibition of clonal growth of KG 1 and HL-60 cell lines and of leukemic cells from two patients was not associated with the presence of a specific cytoplasmic retinoic acid-binding protein. Our study suggests that retinoic acid may prove to be effective in the treatment of human myeloid leukemia. PMID- 6276438 TI - Monocyte and granulocyte-mediated tumor cell destruction. A role for the hydrogen peroxide-myeloperoxidase-chloride system. AB - Human monocytes stimulated with phorbol myristate acetate were able to destroy a T lymphoblast cell target (CEM). Stimulated human granulocytes were also capable of mediating CEM cytotoxicity to a comparable degree as the monocyte. CEM destruction was dependent on the pH and the effector cell number. Both monocyte or granulocyte mediated cytotoxicity were inhibited by the addition of catalase, whereas superoxide dismutase had no inhibitory effect. In addition, CEM were protected from cytolysis by the effector cells by the myeloperoxidase inhibitors, azide and cyanide, or by performing the experiment under halide-free conditions. Glucose oxidase, an enzyme system capable of generating hydrogen peroxide, did not mediate CEM cytotoxicity, while the addition of purified myeloperoxidase dramatically enhanced cytolysis. Hypochlorous acid scavengers prevented CEM destruction by the glucose oxidase-myeloperoxidase-chloride system but neither hydroxyl radical nor singlet oxygen scavengers had any protective effect. These hypochlorous acid scavengers were also successful in inhibiting monocyte or granulocyte-mediated CEM cytotoxicity. Based on these observations we propose that human monocytes or granulocytes can utilize the hydrogen peroxide myeloperoxidase-chloride system to generate hypochlorous acid or species of similar reactivity as a potential mediator of CEM destruction. PMID- 6276440 TI - Sodium channel and sodium pump in normal and pathological muscles from patients with myotonic muscular dystrophy and lower motor neuron impairment. AB - TWO SODIUM TRANSPORT SYSTEMS HAVE BEEN ANALYZED IN THIS WORK: the voltage sensitive sodium channel and the (Na(+), K(+)) ATPase pump. The sodium channel has been studied using a tritiated derivative of tetrodotoxin; the sodium pump has been studied using tritiated ouabain. Properties of interaction of tritiated tetrodotoxin and of tritiated ouabain with their respective receptors were observed in normal human skeletal muscle and in muscles of patients with myotonic muscular dystrophy and with lower motor neuron impairment. Levels of sodium pump and of sodium channels were measured at different stages of membrane purification. Microsomal fractions of normal human muscle have maximal binding capacities for tetrodotoxin of 230 fmol/mg of protein and of 7.4 pmol/mg of protein for ouabain. Dissociation constant for the complexes formed by the tetrodotoxin derivative and by ouabain with their respective receptors were 0.52 nM and 0.55 muM, respectively. In muscles from patients with myotonic muscular dystrophy, the maximal binding capacity for tetrodotoxin, i.e., the number of Na(+) channels was found to be very similar to that found for normal muscle. The maximal binding capacity for ouabain, i.e., the number of Na(+) pumps was three- to sixfold lower than in normal muscle. Dissociation constants for the complexes formed with the tetrodotoxin derivative and with ouabain were the same as for normal muscle. In muscles from patients with lower motor nerve impairment, the maximal binding capacities for tetrodotoxin and for ouabain were twice as high as in normal muscle. Again, dissociation constants for the complexes formed with the tetrodotoxin derivative and with ouabain were nearly unchanged as compared with normal muscle. These results suggest that sodium transport systems involved in the generation of action potentials and/or in the regulation of the resting potential are altered both in myotonic muscular dystrophy and in lower motor neuron impairment. PMID- 6276441 TI - 3,3',5-triiodothyronine administration in vivo modulates the hormone-sensitive adenylate cyclase system of rat hepatocytes. AB - The ability of 10 muM epinephrine or isoproterenol to stimulate cyclic AMP accumulation was decreased in hepatocytes isolated from hyperthyroid (triiodothyronine treated) as compared to euthyroid rats. In the presence of methylisobutylxanthine, epinephrine or isoproterenol-stimulated cyclic AMP accumulation was approximately 65% lower in hyperthyroid as compared with euthyroid rat hepatocytes. The ability of glucagon to stimulate a cyclic AMP response was also decreased in the hyperthyroid state, when assayed in either the absence or presence of a methyl xanthine. The character of the catecholamine stimulated cyclic AMP response was beta adrenergic in both the hyperand euthyroid states. No evidence for an alpha(2) adrenergic mediated component of catecholamine action on cyclic AMP levels was noted. Cyclic AMP phosphodiesterase activity of hepatocyte homogenates was not altered in the hyperthyroid state. Hormone-stimulated, guanine nucleotide- and fluoride-activatable adenylate cyclase activity was reduced in subcellular fractions obtained from hyperthyroid as compared with euthyroid rat hepatocytes. Beta adrenergic receptor binding was reduced approximately 35% and glucagon receptor binding reduced approximately 50% in the hyperthyroid as compared with euthyroid rat hepatocyte membrane fractions. The status of the regulatory components of adenylate cyclase were examined by in vitro treatment of subcellular fractions with cholera toxin. The ability of cholera toxin to modulate adenylate cyclase was not altered by hyperthyroidism. Cholera toxin catalyzed AD[(32)P]ribosylation of hyperthyroid and euthyroid rat hepatocyte proteins separated electrophoretically displayed nearly identical autoradiograms. Studies of the reconstitution of adenylate cyclase activity of S49 mouse lymphoma cyc(-) mutant membranes by detergent extracts from rat hepatocyte membranes, indicated that hyperthyroidism was associated with a reduced capacity of regulatory components to confer fluoride, but not guanine nucleotide activatability to catalytic cyclase. Thyroid hormones regulate the hormone-sensitive adenylate cyclase system of rat hepatocytes at several distinct loci of the system. PMID- 6276442 TI - Intracellular potassium activity in guinea pig papillary muscle during prolonged hypoxia. AB - During prolonged hypoxia, intracellular potassium concentration, [K](i) has been reported to fall by 70% with a concomitant decrease in the calculated potassium equilibrium potential, E(K). Nevertheless, resting membrane potential, V(m), declined only slightly. Because V(m) depolarized very little in relation to the calculated E(K), it was hypothesized that electrogenic Na-K pumping contributed up to 40 mV to V(m) during prolonged hypoxia. To further test this hypothesis we studied what changes prolonged hypoxia makes in the thermodynamically active fraction of cellular potassium, intracellular potassium activity, alpha(K) (i), and how change in alpha(K) (i) affects the relationship between V(m), E(K) and, by inference, the Na-K pump. Using double-barrel K-selective electrodes, V(m) and alpha(K) (i) were measured in quiescent guinea pig right ventricular papillary muscles superfused for 8 h with hypoxic Tyrode's solution. Over the 8-h period both V(m) and alpha(K) (i) decreased. However, the decline in V(m) was paralleled by a decrease in the E(K) calculated from alpha(K) (i). At no time was there hyperpolarization of V(m) beyond E(K). After 8 h the Na-K pump was inhibited by exposing the muscles to 0.1 mM ouabain. The onset of an increase in extracellular potassium activity, measured with a double-barrel electrode, was used to mark the amount of depolarization of V(m) due solely to pump inhibition. After hypoxia, V(m) depolarized 8.4+/-4.4 mV before extracellular potassium activity (alpha(K) (e)) increased. Thus, the decrease in alpha(K) (i) during hypoxia is much less than that reported for [K](i). The parallel decline in V(m) and E(K) and the small depolarization of V(m) with ouabain suggest that after prolonged hypoxia the Na-K pump continues to contribute to V(m), but the amount of this contribution is substantially less than previously reported. PMID- 6276443 TI - Alkaline phosphatase for immunocytochemical labelling: problems with endogenous enzyme activity. AB - Alkaline phosphatase may be used as a label for immunocytochemistry and can be demonstrated in tissue sections using the single step naphthol phosphate method. Endogenous enzyme activity may not be destroyed by fixation in formalin, formol alcohol, Carnoy's or Baker's solutions and should be inhibited before results are assessed. Either Bouin's solution or periodic acid followed by potassium borohydride are satisfactory inhibitor and do not adversely affect immunocytochemical results. PMID- 6276444 TI - Evaluation of a new enzyme-linked immunosorbent assay test for rotavirus antigen in faeces. AB - A new commercial test for the diagnosis of rotavirus gastroenteritis was assessed. With some modifications it compared favourably with electron microscopy and immunofluorescence. PMID- 6276445 TI - Further observations on the cerebellar projections from the pontine nuclei and the nucleus reticularis tegmenti pontis in the rhesus monkey. AB - The projections from the pontine nuclei and the necleus reticularis tegmenti pontis (N.r.t.) onto the flocculus, uvula, and the paramedian lobule were studied with retrograde transport of horseradish peroxidase n the rhesus monkey. The main findings are as follows: There is a conspicuous tendency for labeled cells to occur in numerous discrete clusters in the pontine nuclei after injections of these parts of the cerebellum. There appears to be very limited overlap between pontine cell groups projecting to the flocculus, the uvula, and the paramedian lobule, respectively. The flocculus appears to receive a substantial projection from the pontine nuclei. The projection is almost totally crossed (3% ipsilateral), and arises mainly laterally in the rostral half of the pons but in addition from a minor group dorsomedially. The flocculus receives a bilateral projection (slight contralateral preponderance) from medial and dorsomedial parts of the NRT. The number of labeled cells in the NRT was 13% of the number in the pontine nuclei. the uvula is amply supplied from the pontine nuclei. The projection takes origin throughout the rostrocaudal extent of the pons, from one medial and one dorsolateral region. Labeled cells are found in greatest number dorsolaterally in the rostral half of the pons. In the caudal N.r.t., one medial and one lateral cell group were labeled after injection of the uvula. The number of labeled cells in the N.r.t. was only 4% of the number in the pontine nuclei. Findings with regard to the paramedian lobule confirm and extend earlier observations in the monkey (Brodal, '79, '80). The present results are discussed in relation to HRP studies of the pontocerebellar projection in lower animals. Several possible species differences are noted--for example, with regard to projections to the flocculus. There is some evidence that the pontocerebellar projection is more precisely organized in the monkey than in lower animals. PMID- 6276446 TI - Retinal W-cell projections to the medial interlaminar nucleus in the cat: implications for ganglion cell classification. AB - The perikaryal sizes and retinal distribution of ganglion cells labeled after small iontophoretic injections of horseradish peroxidase (HRP) into the medial interlaminar nucleus (MIN) were studied. Injections were also made into the LGNv and the C-laminae of the dorsal lateral geniculate nucleus (LGNd) for comparison. The results are consistent with suggestions that the MIN contains three approximately vertically oriented laminae which, from medial to lateral, receive their input from, respectively, contralateral nasal, ipsilateral temporal, and contralateral temporal retina. Each MIN lamina receives afferents from two distinct groups of retinal ganglion cells (1) cells with large somas (over 25 micron), coarse primary dendrites, large dendritic trees (500-900 micron in diameter), and coarse axons; (2) cells with medium-sized somas (14-20 micron), medium-caliber primary dendrites, large dendritic trees (350-700 micron), and fine axons. The large cells are clearly Y-cells or alpha cells, and they provide approximately 50% of the retinal input to all layers of the MIN. The medium-sized cells, which provide the remaining 50% of the retinal output in the MIN, are, we argue, W-cells, since they do not differ in soma size, dendritic morphology, axon caliber, or receptive field properties from medium-sized W-cells which project to other thalamic or midbrain structures. These results suggest two phylogenetic trends within the W-cell group: (1) the differentiation of thalamic and midbrain components; and (2) the further differentiation of ipsilateral and contralateral projections within the midbrain component. This latter division corresponds to the distinction between W1 and W2 cells described previously (Rowe and Stone, '77, '80). PMID- 6276448 TI - The organization of binocular cortex in the primary visual area of the rabbit. PMID- 6276447 TI - Afferent projections to the deep mesencephalic nucleus in the rat. AB - Afferent projections to the deep mesencephalic nucleus (DMN) of the rat were demonstrated with axonal transport techniques. Potential sources for projections to the DMN were first identified by injecting the nucleus with HRP and examining the cervical spinal cord, brain stem, and cortex for retrogradely labeled neurons. Areas consistently labeled were then injected with a tritiated radioisotope, the tissue processed for autoradiography, and the DMN examined for anterograde labeling. Afferent projections to the medial and/or lateral parts of the DMN were found to originate from a number of spinal, bulbar, and cortical centers. Rostral brain centers projecting to both medial and lateral parts of the DMN include the ipsilateral motor and somatosensory cortex, the entopeduncular nucleus, and zona incerta. at the level of the midbrain, the ipsilateral substantia nigra and contralateral DMN likewise project to the DMN. Furthermore, the ipsilateral superior colliculus projects to the DMN, involving mainly the lateral part of the nucleus. Afferents from caudal centers include bilateral projections from the sensory nucleus of the trigeminal complex and the nucleus medulla oblongata centralis, as well as from the contralateral dentate nucleus. The projections from the trigeminal complex and nucleus medullae oblongatae centralis terminate in the intermediate and medial parts of the DMN, whereas projections from the contralateral dentate nucleus terminate mainly in its lateral part. In general, the afferent connections of the DMN arise from diverse areas of the brain. Although most of these projections distribute throughout the entire extent of the DMN, some of them project mainly to either medial or lateral parts of the nucleus, thus suggesting that the organization of the DMN is comparable, at least in part, to that of the reticular formation of the pons and medulla, a region in which hodological differences between medial and lateral subdivisions are known to exist. PMID- 6276449 TI - Localization of synaptic and nonsynaptic nicotinic-acetylcholine receptors in the goldfish retina. AB - The localization of nicotinic-cholinergic receptors in the inner plexiform layer (IPL) of goldfish retina was studied by electron microscopic analysis of the binding pattern of a conjugate or horseradish peroxidase and alpha bungarotoxin (HRP-alpha BTx). Specific HRP reaction product (blockade by 1mM curare) was found at both synaptic and nonsynaptic sites. Synaptic binding sites for HRP-alpha BTx, which accounted for only 16% of the total specific reaction product sites, always involved an amacrine process as the presynaptic element, whereas amacrine, ganglion, and bipolar cells could be post-synaptic elements at labeled synapses. Only 17.5% of the total number of amacrine synapses were labeled by HRP-alpha BTx. Labeled synapses showed the same distribution in the IPL as unlabeled synapses: bimodal for amacrine-to-bipolar synapses with peak concentrations at the 20% and 80% layers and unimodal for amacrine-to-nonbipolar synapses with a peak concentration at the 60% layer. Nonsynaptic binding sites for HRP-alpha BTx (84% of total) were seen on the dendrites of ganglion, amacrine, and bipolar cells. The distribution of the nonsynaptic sites in the IPL largely accounts for the trilaminar binding pattern of 125I-alpha BTx as observed in light microscopic autoradiographs. If, as appears likely, the distribution of synapses is the relevant variable in determining the sites of neuronal interaction for a given transmitter system, then this study further illustrates the importance of distinguishing synaptic from nonsynaptic binding when using receptor-ligand probes to localize sites of chemical synaptic transmission. PMID- 6276450 TI - Intrinsic connections and architectonics of posterior parietal cortex in the rhesus monkey. AB - By means of autoradiographic and ablation-degeneration techniques, the intrinsic cortical connections of the posterior parietal cortex in the rhesus monkey were traced and correlated with a reappraisal of cerebral architectonics. Two major rostral-to-caudal connectional sequences exist. One begins in the dorsal postcentral gyrus (area 2) and proceeds, through architectonic divisions of the superior parietal lobule (areas PE and PEc), to a cortical region on the medial surface of the parietal lobe (area PGm). This area has architectonic features similar to those of the caudal inferior parietal lobule (area PG). The second sequence begins in the ventral post/central gyrus (area 2) and passes through the rostral inferior parietal lobule (areas PG and PFG) to reach the caudal inferior parietal lobule (area PG). Both the superior parietal lobule and the rostral inferior parietal lobule also send projections to various other zones located in the parietal opercular region, the intraparietal sulcus, and the caudalmost portion of the cingulate sulcus. Areas PGm and PG, on the other hand, project to each other, to the cingulate region, to the caudalmost portion of the superior temporal gyrus, and to the upper bank of the superior temporal sulcus. Finally, a reciprocal sequence of connections, directed from caudal to rostral, links together many of the above-mentioned parietal zones. With regard to the laminar pattern of termination, the rostral-to-caudal connections are primarily distributed in the form of cortical "columns" while the caudal-to-rostral connections are found mainly over the first cortical cell layer. PMID- 6276451 TI - The thalamic connectivity of the primary motor cortex (MI) in the raccoon. AB - The purpose of this study was to determine the pattern of thalamic projections of the primary motor cortex (MI) in the raccoon, a carnivore species noted for neural specialization of sensorimotor function. Following electrophysiological identification of circumscribed regions of MI, injections of horseradish peroxidase (HRP) or HRP combined with tritiated amino acids were made in 15 animals. Labeled thalamic cells were found predominantly in the ventral lateral nucleus (VL). For a given cortical injection site within MI, labeled neurons in VL formed a crescent-shaped band which extended in a dorsoventral direction when viewed in coronal sections. These bands were topographically organized. Following an injection of the MI hindlimb area in the medial part of the posterior cruciate gyrus, both retrogradely labeled neurons and anterograde label formed a thin band at the lateral edge of VL while an injection of the MI face area in the lateral part of the anterior cruciate gyrus resulted in both anterograde and retrograde label in medial VL and the principal division of the ventromedial nucleus (VMp). An injection of the MI forepaw area localized to the rostral and central part of the posterior cruciate gyrus resulted in a band of labeled neurons occupying the dorsal extent of VL and continuing into the ventrolateral aspect of the ventral anterior nucleus (VA). In contrast, an injection of the MI forepaw area which was localized to the caudal extent of the posterior cruciate gyrus resulted in a wide and diffuse band of labeled neurons and anterograde label in the ventral portion of VL. All injection of MI produced cell labeling in the paracentral nucleus (PC) and the central lateral nucleus (CL) of the intralaminar group. These results demonstrate that VL is the primary thalamic dependency of MI in the raccoon. Labeled cells were not observed in the ventrobasal complex. The MI pattern of thalamic connectivity observed in the present study suggests that while differences exist in the regional specialization of sensorimotor structures among species, there appears to be little variation in the overall organization of thalamocortical relations. PMID- 6276452 TI - Membrane specializations in the outer plexiform layer of the turtle retina. AB - The internal organization of the plasma membrane at specialized contacts in the outer plexiform layer of the turtle, Pseudemys scripta elegans, was analyzed with the aid of the freeze-fracturing technique. In the invaginating synapse of cone pedicles the plasma membrane of the photoreceptor ending contains an aggregate of P-face particles, images of synaptic vesicle exocytosis, and rows of forming coated vesicles which are arranged in sequence from apex to base of the synaptic ridge. Thus, freeze-fracturing provides positive evidence that the synaptic ridge represents the active zone at the surface of the photoreceptor endings. Horizontal cell processes of dyads and triads have an aggregate of P-face particles opposite the apex of the ridge, but lack images of vesicle exocytosis. Deep-etching and rotary-shadowing demonstrate that an array of minute protrusions decorates the true outer surface of the horizontal cell membrane at the site of the intramembrane particle aggregate. The membrane of the invaginating bipolar dendrite is unspecialized. At basal junctions, the cone pedicle membrane has a sparse complement of P-face particles, but images of vesicle exocytosis are absent. The adjoining bipolar membrane is characterized by a prominent aggregate of E-face particles, often arranged in an orthogonal lattice. The freeze-fracture profile therefore suggests the existence of (1) a sign-conserving cone-to horizontal cell synapse; (2) a sign-inverting synapse between cones and invaginating bipolar dendrites; and (3) a sign-conserving synapse between cones and bipolar dendrites at basal junctions. No freeze-fracture evidence was found for a horizontal-to-cone or horizontal-to-bipolar cell synapse within the synaptic invaginations. PMID- 6276453 TI - Organization of the hand area in the primary somatic sensory cortex (SmI) of the prosimian primate, Nycticebus coucang. PMID- 6276454 TI - Selective associations in one-day-old rats: taste-toxicosis and texture-shock aversion learning. AB - One-day-old rats learned aversions to a novel taste paired with lithium-induced distress and to a tactile stimulus paired with brief electric shocks. However, aversions did not develop when taste was paired with shock or when the tactile stimulus was paired with lithium treatment. The aversions occurred only when lithium treatment immediately followed taste exposure and when shock was concurrent with exposure to the tactile stimulus. These findings indicate that selective associations in aversion learning are mediated by innate mechanisms that govern conditioning in the absence of extensive ontogenetic experience. The present research also shows that selective associations are sufficient for the occurrence of long-delay learning. PMID- 6276455 TI - Specific and nonspecific changes in visual evoked potentials during eyelid conditioning in the rabbit (Oryctolagus cuniculus). AB - Eye blinks were conditioned to either the visual or nonvisual element of a compound conditioned stimulus. The visual element consisted of a series of electrical pulses to the optic chiasma, and the averaged evoked potentials (AEPs) produced by this stimulus were recorded in the dorsal lateral geniculate nucleus and the visual cortex. The initial surface positive component of the cortical AEP was enhanced only when the eye blinks were conditioned to the visual stimulus, an effect that cannot be attributed to nonspecific mechanisms. The "postsynaptic" component of the geniculate AEP was also enhanced, but this occurred regardless of whether the eye blinks were conditioned to the visual or nonvisual stimulus, an effect that appears to be entirely nonspecific. Data from recovery cycles indicate that this enhancement effect cannot be attributed to an inhibition of inhibitory interneurons in the lateral geniculate nucleus. PMID- 6276456 TI - Carbonate apatites from aqueous and non-aqueous media studied by ESR, IR, and X ray diffraction: effect of NH4+ ions on crystallographic parameters. AB - Carbonate-containing apatites prepared from aqueous and non-aqueous media were investigated by ESR, IR, and X-ray diffraction. A typical asymmetric signal at g = 2.0, which many workers have observed in irradiated calcified tissues, is interpreted in terms of CO3(3)- and Co-2 and the trapped electron center, The CO3(3)- and CO2- ions originated from the carbonate which substitutes for the phosphate ion in the apatite crystalline structure. From the comparison of the data between the carbonate apatites prepared under the presence of Na+ and NH4+ ions, it is suggested that NH4+ ions contribute to the crystallographic parameters, either by substituting in the structure or by suppressing the CO3 incorporation into the structure, or by both. The production of the CO2- radical ion could be accounted for based on the concept of hydrogen bond between the oxygen atom of CO2(3)- and the hydrogen atom of NH4+. PMID- 6276457 TI - Enzyme patterns of glycolysis in rabbit gingiva and effects of pH on the patterns. PMID- 6276458 TI - Cefotaxime in treatment of meningitis and ventriculitis? Evaluation of drug concentrations in human cerebrospinal fluid. AB - In three groups of patients levels of cefotaxime in serumand cerebrospinal fluid were determined. Therapeutic value and efficacy are discussed in meningitis patients. Nine concentrations of cefotaxime in lumbar and ventricular CSF out of 19 in a group of seven neurosurgical patients with mild to moderate impairment of the blood-CSF-barrier were higher than 0.5 micrograms/ml. In seven determinations in a second group of six patients with no or very little dysfunction of the blood cerebrospinal-fluid barrier only twice cefotaxime was not detectable in lumbar CSF. Concentrations of cefotaxime in 25 determinations of lumbar or ventricular CSF in six patients with bacterial meningitis ranged from 1.1 micrograms/ml to 19.2 micrograms/ml. Treatment with cefotaxime alone was successful in a patient with E. coli meningitis and ventriculitis after infection of a ventriculo-atrial shunt and in another patient with pneumococcal meningitis and penicillin allergy. The other four patients with bacterial meningitis were treated successfully by antibiotics including cefotaxime. PMID- 6276459 TI - Changes in angiotensin-converting enzyme activity and angiotensin I level in asthmatic and healthy children after submaximal physical work. AB - The changes in angiotensin-converting enzyme activity and serum angiotensin I levels have been studied in 16 controls subjected to submaximal physical work. Baseline (Pre-exercise) angiotensin I levels were identical in both groups. Physical exercise caused an elevation that was more marked in the asthmatic group than in the control group. The activity of serum angiotensin-converting enzyme differed in the two groups even before physical exercise, the asthmatic children having exhibited an activity level significantly lower than that of the healthy controls. after submaximal work, the enzyme activity increased in healthy subjects but decreased in asthmatic children. PMID- 6276460 TI - Biologic aspects of falls and mobility limitations in the elderly. PMID- 6276462 TI - Multifocal benign granular cell tumor of the stomach. PMID- 6276461 TI - Isoproterenol responsiveness and myocardial beta-adrenergic receptors in young and old rats. AB - To elucidate the association of beta-adrenergic receptors with decreased myocardial catecholamine responsiveness in aging, we investigated the chronotropic response to isoproterenol and myocardial beta-adrenergic receptors in Fisher 344 rats of 3, 12, and 24 months of age. Heart rate response to isoproterenol (50 mug/kg, subcutaneously) was greatest in 3-month-old animals, both in maximum heart rate achieved and increment over baseline. Twelve-month-old animals had significantly lower maximal hear rates (p less than .025) and increment over pretreatment, basal minus isoproterenol-stimulated heart rates (p less than .005), when compared with 3-month-old animals. Twenty-four-month-old rats were less responsive than either 12- or 3-month-old animals both with respect to maximal rates (p less than .05, .005, respectively) and increments (p less than .005, .001, respectively). There were no changes in the density or affinity of myocardial, lymphocytic, or pulmonary beta-adrenergic receptors as measured by [3H]-dihydroalprenolol binding in the three age groups investigated. Our data are consistent with the concept that the beta-adrenergic stimulatory pathway is impaired with age. The demonstration of unaltered myocardial beta adrenergic receptor number and affinity with age suggests that myocardial catecholamine responsiveness is impaired at a level other than the beta adrenergic receptor site. PMID- 6276463 TI - [Cholinergic mechanisms in the retina of rats]. AB - In rats with extra- or intracranial dissection of the visual pathway the activities of acetylcholinesterase (AChE) and choline acetyltransferase (ChAc) were determined in retina, optic nerve and optic tract quantitatively by bio- and histochemical methods after one or five weeks of survival time. One week after intracranial (chiasma-near) dissection of the optic nerve an initial ipsilateral enhancement in the enzyme activity of AChE was found histochemically in retina layers and optic nerve. Five weeks following the dissection significant losses in AChE activity could be determined in the optic nerve, less pronounced in the retina. However, the activity of ChAc in the retina never was changed, neither after short nor after long survival times. Therefore, the high activity of both enzymes connected with cholinergic transmission normally present in different retinal layers must belong to local neurons, probably amacrine cells. To elucidate the question of the existence of a centrifugal cholinergic innervation of the retina one eye was enucleated. One week after operation a significant increase in the activity of AChE and ChAc was found in central parts of the dissected optic nerve and in the opposite optic tract indicating the existence of centrifugal cholinergic fibers in the optic nerve, but no evidence for the origin of these fibres could be obtained. The possible role of a central cholinergic control of the visual information processing already in retinal level is discussed in analogy to similar results obtained in the olfactory bulb. PMID- 6276464 TI - Microbiological studies on hamburgers. AB - One hundred and eighty-two raw, 112 pre-cooked and 750 cooked hamburgers composed mainly of beef or beef and pork were subjected to microbiological examination.Raw hamburgers gave total bacterial counts from 10(6) to 10(8) per g, counts of Enterobacteriaceae from 10(4) to 10(6) per g, of Escherichia coli from 10(3) to 10(5), of group D streptococci from 10(2) to 10(4), of Staphylococcus aureus from 3 to 10(2) and of Clostridium perfringens less than 10 bacteria per g. Of the samples, 32% contained salmonellas; the highest most probable number was 10(2) per g but most estimates were below 1 per g. Corresponding figures for the pre cooked samples were 2-3 log cycles lower, and only one sample contained salmonella. Yersinia enterocolitica was not isolated from any raw or pre-cooked sample.Three hundred and ninety-five of the cooked hamburgers were prepared by grilling raw hamburgers for between 2 and 5.5 min. These gave total bacterial counts from 10(5) to 10(7) per g, and counts of Enterobacteriaceae from 10(2) to 10(5) per g. Of the samples, 9.4% contained salmonellas, always in numbers below 1 per g. The remaining 355 cooked hamburgers were prepared from samples pre cooked for 10 min at 80 degrees C. Some were grilled and some fat fried. The total bacterial counts were from 10(3) to 10(5) per g, and counts of Enterobacteriaceae below 10(2) per g. Salmonellae, again in small numbers only, were recovered from 3.5% of samples.When hamburgers were artificially contaminated with Salmonella typhimurium it took 5.5 min on a commercial grill, 2.25 min frying in a frying pan and 1.75 min on a household grill to reliably reduce the salmonella count one hundredfold. This means that at many vending places hamburgers are often cooked for too short a time.D-values were determined for S. typhimurium in hamburger meat at 50, 55, 60, 65 and 70 degrees C, these values were 7.1, 5.1, 1.2, 0.9 and 0.6 min respectively. It can be concluded that the heating action in the centre of the hamburgers will take place more slowly than in the hamburger as a whole, and that the time between cooking and consumption is very important in reducing the microbial load to acceptable levels.Pre-cooking (10 min at 80 degrees C in a water bath) gives a reduction in the numbers of salmonella of about 4 x 10(3), after which cooking gives a further reduction as mentioned above. PMID- 6276465 TI - A reappraisal of serotype factors 4, 5 and 6 of Bordetella pertussis. AB - Twelve strains of Bordetella pertussis, of various serotypes, were tested by agglutination with serial dilutions of six antisera which were thought by their donor to be monospecific for six distinct agglutinogens. The first three sera were shown, by agglutination of the twelve donated strains and of standard pertussis serotypes, to be monospecific for the three well-recognized major agglutinogens (factor 1, factor 2, factor 3). The 'factor 4' serum gave weak agglutination, but only of strains that possessed factor 2, though not all of such strains. This suggested that it may be a distinct minor component of factor 2. The 'factor 5' serum was very weak, and gave agglutination only with a few of the factor 2 strains. The titres of the 'factor 6' serum were very closely parallel to those of the factor 3 serum, and it may be inferred that these 'factors' probably constitute a single antigenic entity. Seven of the twelve donated strains were shown to be mixtures of two or more serotypes, offering an explanation for discrepancies in serological tests involving the use of these strains on different occasions. It seems likely that 'factors 5 and 6' do not actually exist; and, since 'factor 4' is only a minor component of factor 2, any protective role that it may have would probably be covered by the presence of factor 2 in vaccine. PMID- 6276466 TI - Microinjection of macromolecules into normal murine lymphocytes by cell fusion technique. I. Quantitative microinjection of antibodies into normal splenic lymphocytes. AB - Human erythrocyte ghosts loaded with various kinds of protein molecules were fused with mouse splenic lymphocytes by means of polyethylene glycol supplemented with poly-L-arginine and dimethylsulfoxide. This fusion method made quantitative microinjection of IgG and other proteins into intact lymphocytes possible. The injection itself did not alter cell viability, and lymphocytes given protein molecules retained intact response activity when they were stimulated with mitogens. Rabbit anticyclic AMP was purified by affinity chromatography and injected into lymphocytes. Antibody activity in the cell lysates was measured by using 125I-labeled cyclic AMP as an antigen, and it was shown that antibody molecules were quantitatively injected and immunologically active in the cells. Antigen binding activity of anti-cyclic AMP antibodies in the nonstimulated lymphocytes was stable and intact even 24 hr after microinjection, whereas the activity rapidly decreased in mitogen-stimulated lymphocytes, indicating that some immunologic or enzymatic mechanisms for inactivating antibodies were induced in mitogen-stimulated cells. Furthermore, microinjection of anti-cyclic AMP markedly enhanced the proliferative responses of lymphocytes to mitogens such as Con A or LPS and reversed the effect of a known elevator of intracellular cyclic AMP. These observations have implications for the role of cyclic AMP in early lymphocyte activation events. PMID- 6276467 TI - Activation of suppressor T cells by autologous lymphoblastoid cells: a mechanism for feedback regulation of immunoglobulin synthesis. PMID- 6276468 TI - Effects of vitamin K on human neutrophil function. PMID- 6276469 TI - Rapid large scale fractionation of lymphoid cells by velocity sedimentation at unit gravity, using Percoll as separation medium. AB - Available methods for separation of cells by velocity sedimentation at unit gravity allow a maximum cell load of 2 x 108 lymphoid cells. Here we describe a method, based on the 'tilting' procedure of Bont et al. (1979), which allows separation of up to 4.5 x 109 lymphoid cells, while retaining full separation and rapidity. Percoll instead of Ficoll was used as gradient medium. The relationship between cell sedimentation velocity and cell volume was analyzed by two different methods, i.e., electronic cell sizing and diameter measurements of cytocentrifuged cells. PMID- 6276470 TI - Separation of human peripheral blood monocytes on continuous density gradients of polyvinylpyrrolidone-coated silica gel (Percoll). AB - A standardized, reproducible two-step method for separation of human peripheral blood monocytes on continuous Percoll gradients has been developed. The first step involves separation of mononuclear cell on Percoll of density 1.075 g/ml and the second step separation of monocytes from lymphocytes on a continuous Percoll gradient with a starting density of 1.075 g/ml for the formation of the gradient. The average yield during a 10 month period of daily routine use has been 74 +/- 17% (mean +/- 1 S.D.), and the average purity 63 +/- 10%. Ninety to 95% of the monocytes are viable after separation as judged from trypan blue exclusion and by ingestion of latex particles and sensitized sheep erythrocytes. The separation takes about 3 h and the total number of monocytes obtained from 40 ml of blood is in the range of 10-15 x 106. The procedure has been reliable with 3-4% separation failures, mainly due to bacterial or fungal growth in Percoll suspension or media. The contaminating cells are exclusively lymphocytes, predominantly T lymphocytes (90-95%), when citrate is used as anticoagulant. Heparin can not be used as anticoagulant, as there appears to be a dose-dependent formation of thrombocyte aggregates which contaminate the monocytes, and result in poor separation. PMID- 6276471 TI - Malignant pulmonary adenomatosis. PMID- 6276472 TI - Decreased collagenase production by regional fibroblasts cultured from skin of a patient with connective tissue nevi of the collagen type. AB - A 10-yr-old female presented with cerebriform tumors covering the plantar surfaces of both feet. Histologically, the lesions consisted of thick collagen fibers and the content of collagen per surface area of skin was increased about 8 fold. Examination of the collagen by SDS-polyacrylamide gel electrophoresis, after limited pepsin proteolysis, showed that the lesions consisted almost exclusively of type I collagen, the predominant collagen type in human skin. Thus, a diagnosis of connective tissue nevi of the collagen type was made. Fibroblast cultures were established from the affected and normal-appearing areas of the skin, and examined for the rate of collagen synthesis, production of collagenase and growth kinetics of the cells. Cell cultures derived from the lesion and from control skin synthesized procollagen at the same rate and in a normal type I/type III procollagen ratio. However, the production of enzymatically active and immunologically detectable collagenase was reduced by 70 82% in the cultures derived from the lesion as compared to controls (p less than 0.005). Fibroblasts derived from the lesions also displayed a mean population doubling time of 1.17 +/- 0.08 days compared to 1.83 +/- 0.24 and 1.92 +/- 0.09 days for control cell strains and cells derived from normal skin of the patient, respectively (p less than 0.025). These results suggest that the excessive deposition of collagen in this case may have resulted from decreased local degradation of collagen. Enhanced proliferative capacity of the regional fibroblasts may have contributed to the accumulation of collagen in these lesions. PMID- 6276473 TI - Correlation between human papillomavirus (HPV) type and histology of warts. AB - Forty warts from different patients and of different clinical type were examined histologically and virologically. Eight lesions were found to be associated with human papillomavirus type 1 (HPV 1), 15 tumors were induced by HPV 2, HPV 3 was detected 4 times, HPV 4 twice, and HPV 6 eleven times. HPV 3, HPV 4, and HPV 6 induced warts revealed a correlation between histology and virus type. They are characterized by the so called "edematous type clear cells". In HPV 3 associated flat warts pycnotic nuclei were mainly localized in the center of large vacuoles. In genital warts sickle shaped nuclei were pushed to the margin of the vacuolized cells. The histology of HPV 1 and HPV 2 induced warts was more heterogenous. With one exception HPV 1-induced lesions represented typical myrmecia warts, varying in the number and shape of inclusion bodies. HPV 2 associated common warts, however, revealed 3 very distinct histologic features: (1) Inclusion wart typical for HPV 1, (2) Classical common wart with marked condensation of keratohyalin granules, (3) Warts with extreme vacuolization of squamous and granular cells leading to a honeycomb-like picture. PMID- 6276474 TI - Measurement of cutaneous inflammation: estimation of neutrophil content with an enzyme marker. AB - We examined the hypothesis that myeloperoxidase (MPO), a plentiful constituent of neutrophils, might serve as a marker for tissue neutrophil content. To completely extract MPO from either neutrophils or skin, hexadecyltrimethylammonium bromide (HTAB) was used to solubilize the enzyme. With this detergent treatment, 97.8 +/- 0.2% of total recoverable MPO was extracted from neutrophils with a single HTAB treatment; 93.1 +/- 1.0% was solubilized with a single treatment of skin. Neutrophil MPO was directly related to neutrophil number; with the dianisidine H2O2 assay as few as 10(4) neutrophils could be detected. The background level of MPO within uninflamed tissue was 0.385 +/- 0.018 units per gram of tissue, equivalent to only 7.64 +/- 0.36 X 10(5) neutrophils. In experimental staphylococcal infection, skin specimens contained 34.8 +/- 3.8 units MPO per gram, equivalent to 8.55 +/- 0.93 X 10(7) neutrophils. These studies demonstrate that MPO can be used as a marker for skin neutrophil content: it is recoverable from skin in soluble form, and is directly related to neutrophil number. Further, normal skin possesses a low background of MPO compared to that of inflamed skin. PMID- 6276475 TI - Pig skin epidermal calmodulin: effect on calmodulin deficient phosphodiesterase. AB - Calmodulin, a calcium-dependent modulator protein, is known to mediate a great number of Ca++-dependent processes in various tissues. Although it was originally described as a protein activator of cyclic nucleotide phosphodiesterase, the sensitivity of phosphodiesterases to this compound are suggested to be variable from tissue to tissue. In order to determine whether there was calmodulin-like activity in pig skin epidermis and to see its relationship to epidermal phosphodiesterase, we used an established calmodulin deficient phosphodiesterase system prepared from bovine heart. Calmodulin deficient phosphodiesterase prepared from bovine heart was markedly stimulated by the addition of pig skin (epidermal) boiled extract in the presence of calcium. Boiled skin extract alone had only little phosphodiesterase activity by itself. This effect of boiled skin extract on bovine heart phosphodiesterase was inhibited by the addition of EGTA, a divalent metal ion chelator of relative Ca++ specificity. At a fixed concentration of EGTA, increasing the Ca++ concentration counteracted the effect of EGTA. Pure pig skin epidermis (separated by trypsinization, NaBr, CaCl2 sucrose or NH4Cl treatment) was also shown to have heat-stable calmodulin activity. In contrast to the bovine heart phosphodiesterase, epidermal phosphodiesterase was only partially inhibited when Ca++ was removed by EGTA. The addition of boiled skin extract on the crude extract of epidermal phosphodiesterase had minimal effect on the enzyme activity. Overall results indicate that although pig skin epidermis contains significant amount of calmodulin, the regulation of phosphodiesterase may not be the main biological activity of epidermal calmodulin. PMID- 6276476 TI - Adhesion of Entamoeba histolytica trophozoites to monolayers of human cells. AB - The adhesion of radiolabeled trophozoites of Entamoeba histolytica to monolayers of a human intestinal epithelial cell line was found to be dependent on time, temperature, and concentration. Adherence seems to be mediated by a carbohydrate binding protein (lectin), which was previously found in trophozoites of E. histolytica. The adherence of the trophozoites was found to be pH-dependent and was inhibited by several N-acetyl-glucosamine-containing glycoconjugates, such as bacterial peptidoglycan, chitin, and IgA; similar results were obtained with the isolated lectin. Furthermore, the semipurified amoebic lectin and wheat germ agglutinin, which have similar sugar specificities, competed with the intact amoeba for receptor sites on the epithelial cells. In addition, both sera from patients with amoebiasis and an IgG fraction from these sera inhibited lectin activity and the adherence of the trophozoites. PMID- 6276477 TI - Resistance of neonatal human lymphoid cells to infection by herpes simplex virus overcome by aging cells in culture. AB - Mononuclear leukocytes freshly harvested from human umbilical cord blood were refractory to infection by herpes simplex virus (HSV) type 1, whereas cells aged in vitro for a week produced 10- to 100-fold or greater more virus. Increased sensitivity of aged cells to infection by HSV was not dependent on active DNA synthesis by the cells at the time of exposure to HSV. The aging phenomenon was not due to the outgrowth of a specific cell subpopulation that is uniquely susceptible to HSV because increased sensitivity to HSV type 1 occurred in a variety of cell subpopulations enriched or depleted of T lymphocytes, phagocytic cells, or adherent cells. About 25% of aged cells contained virions detectable by electron microscopy, considerably more cells than formed infectious centers. These virions were seen in large activated lymphocytes which are the principal cell subpopulation present in the aged cultures, but virions were also seen in small lymphocytes. PMID- 6276478 TI - From the centers for disease control. Improved accuracy and specificity of forecasting deaths attributed to pneumonia and influenza. PMID- 6276479 TI - [Intermittent hepatic artery occlusion for nonresectable liver cancer (author's transl)]. PMID- 6276480 TI - Degradation and oxidation of methionine enkephalin by human neutrophils. AB - Met5-enkephalin, tyr-gly-phe-met, is an endogenous pentapeptide, with morphine agonist activity. In this study, we demonstrated that met5-enkephalin was degraded with the release of tyrosine by resting human PMN, whereas it was degraded as well as oxidized to its sulfoxide derivative, met5-(O)-enkephalin, by phagocytosing PMN. PMN also degraded met5-(O)-enkephalin but to a lesser extent. Bacitracin at 1 gm/L inhibited the degradation and oxidation of met5-enkephalin without affecting the production of superoxide and viability of PMN. The oxidation of met5-enkephalin by phagocytosing PMN was inhibited by catalase or NaN3 but not by SOD. This suggests that the oxidation of met5-enkephalin by phagocytosing PMN was, at least in part, dependent on the MPO system (MPO-H2O2 halide). Using purified canine MPO, we further demonstrated that MPO-H2O2-CI- oxidized met5-enkephalin to met5-(O)-enkephalin. The MPO-mediated oxidation of met5-enkephalin was inhibited by methionine but not by methionine sulfoxide, tyrosine, glycine, or phenylalanine, confirming that it was the methionine moiety of met5-enkephalin which was oxidized. Since both the sulfoxide derivative and the degradation products met5-enkephalin have reduced opiate agonist activity, oxidation and degradation of met5-enkephalin by PMN may contribute to the pain at the site of inflammation. (J Lab Clin Med 99:418, 1982.) PMID- 6276482 TI - High resolution nuclear magnetic resonance spectroscopy of bile salts: individual proton assignments for sodium cholate in aqueous solution at 400 MHz. AB - The 400 MHz 1H-nuclear magnetic resonance spectrum of sodium cholate in dilute aqueous solution has been successfully resolved using a combination of decoupling, partial relaxation, and decoupled partial relaxation techniques. The individual carbon resonances in the 13C-NMR spectrum of sodium cholate have also have assigned. Assignments of individual methylene protons were made by consideration of the molecular structure of sodium cholate and the expected couplings and 1H-nuclear Overhauser enhancement experiments. Verification of the assignments of the methine protons was made by application of single frequency 1H decoupled 13C-NMR. Variation of pH* from 6.0 to 11.0 did not alter the individual chemical shifts except for those between 2.12 delta and 2.30 delta, originating from the protons on the C23 position adjacent to the ionizable carboxyl group. The chemical shifts of the proton resonances were independent of concentration below 5 mM. Above 10 mM (micellar region), the proton chemical shifts were altered slightly and some band broadening occurred. These data are consistent with the formation of small micellar aggregates (up to N = 4) of cholate molecules. PMID- 6276481 TI - 3-hydroxy-3-methylglutaryl coenzyme A reductase from rat intestine: subcellular localization and in vitro regulation. AB - The subcellular localization of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase in rat intestine was reinvestigated. Highly enriched fractions of endoplasmic reticulum and mitochondria were prepared from mucosal cells. The highest specific activity of HMG-CoA reductase was located in the endoplasmic reticulum fraction with recovery of 25% of the total activity. The mitochondria had low specific activity and low recovery of reductase activity relative to whole homogenate (2-5%). Despite attempts to maximize cell lysis, much of the activity (about 60%) was recovered in a low speed pellet which consisted of whole cells, nuclei, and cell debris as determined by light microscopy. Taken together, the evidence strongly suggests that much of the cellular HMG-CoA reductase activity is present in the endoplasmic reticulum fraction and that mitochondria have little or no intrinsic HMG-CoA reductase. The in vitro regulation of intestinal microsomal HMG-CoA reductase was studied. The intestine possesses a cytosolic HMG-CoA reductase kinase-phosphatase system which appears to be closely related to that present in the liver. Intestinal reductase activity in microsomes prepared from whole mucosal scrapings was inhibited 40-50% by the presence of 50 mM NaF in the homogenizing buffer. It was less susceptible to the action of the kinase than liver reductase. The effects of NaF were reversed by incubation with partially purified intestinal or liver phosphatases. These results suggest that the kinase-phosphatase system could play a role in the regulation of intestinal sterol and isoprene synthesis in vivo. PMID- 6276483 TI - Measurement of receptor-independent lipoprotein catabolism using 1,2 cyclohexanedione-modified low density lipoprotein. AB - The utility of 1,2 cyclohexanedione-modified low density lipoprotein (CHD-LDL) as a marker for the measurement of receptor independent LDL catabolism has been assessed by examining its metabolic properties in cultured human fibroblasts and in rabbits. Cell culture studies showed that the inhibition of high affinity membrane receptor binding produced by the modification could be partially reversed by prolonged incubation of the CHD-LDL at 37 degrees C. Pre-exposure of the complex to alkaline pH (pH 10.5) prevented this and yielded a product that was apparently stable. Despite its regained ability to bind to the fibroblast receptor, 125I-labeled CHD-LDL incubated at 37 degrees C for 24 hr either in vivo or in vitro was removed from rabbit plasma in the same manner as freshly prepared 131I-labeled CHD-LDL and as 131I-labeled CHD-LDL that had been treated at pH 10.5. However, its plasma clearance was significantly faster than that of reductively methylated LDL. We believe that this may result from differential catabolism of these modified lipoproteins rather than from susceptibility of the CHD-LDL to receptor-directed catabolism. PMID- 6276484 TI - Suppression of luteinizing hormone induced by adrenocorticotrophin in healthy women. AB - The effect of ACTH on serum concentrations of LH and FSH was studied in six healthy women in the follicular phase of the menstrual cycle, in six patients (five men, one woman) with adrenocortical insufficiency. In healthy women the i.v. administration of synthetic 1-24 ACTH (0.25 mg) induced a fall in serum concentrations of LH from 11.1 +/- 1.2 (S.D.) to 7.8 +/- 0.6 i.u./l (P less than 0.005) after 30 min and to 8.2 +/- 0.7 i.u./l after 60 min. Upon continuous infusion of 1-24 ACTH (0.25 mg i.v., t = 480 min) LH fell to 6.7 +/- 0.9 i.u./l (P less than 0.005) in healthy women and to 6.1 +/- 3.7 i.u./l (basal, 8.7 +/- 3.9 i.u./l) in healthy men. In patients with adrenocortical insufficiency serum concentrations of LH were unchanged by 1-24 ACTH. Serum concentrations of FSH were not altered by 1-24 ACTH in any of the three groups. It is suggested that the effect of ACTh on LH secretion is healthy women is mediated by the acute rise of endogenous cortisol concentrations. PMID- 6276485 TI - Steroidogenic and glycolytic responses to nucleotides, nucleosides and steroids in rodent adrenal glands: opposing, species-dependent effects of cyclic GMP. PMID- 6276486 TI - Effects of alpha-melanocyte-stimulating hormone and [8-arginine]-vasotocin upon melanogenesis in hair follicle melanocytes in vitro. AB - alpha-Melanocyte-stimulating hormone (alpha-MSH) has been shown to act directly on the mammalian melanocyte in short-term cultures of hair follicles obtained from the Siberian hamster. Melanogenesis was stimulated through an increase in tyrosinase activity which resulted in an increase in melanin production. The response of hair follicle melanocytes to alpha-MSH occurred only in follicles taken from moulting animals, implying that they show a discontinuous expression of MSH receptors during the hair follicle growth cycle. Synthetic 1-24 ACTH had no effect on melanogenesis regardless of whether the follicles came from moulting or non-moulting animals. The pineal peptide, [8-arginine]-vasotocin (AVT), inhibited melanin production without a concomitant decrease in tyrosinase activity. In this respect AVT resembled melatonin, although AVT showed a potency ratio of less than half on a molar basis. The action of AVT, like that of melatonin, must ultimately be on some post-tyrosinase step in melanin biosynthesis. In these hair follicle melanocytes AVT seems to bind to specific receptors since neither of the closely related peptides, oxytocin and [8 arginine]-vasopressin, displayed any activity in our culture system. PMID- 6276487 TI - Fetal testosterone in mice: effect of gestational age and cannabinoid exposure. AB - The effect of increasing gestational age and maternal exposure to cannabinoids on body weight, ano-genital distance and androgen concentration in fetal mice was examined. Body weight increased in both male and female fetuses from days 16 to 18 (the presence of a vaginal plug was considered to indicate day 1 of pregnancy), while ano-genital distances tended to increase faster in male than in female fetuses. The concentration of testosterone increased with age in fetuses of either sex. However, at day 16, there was a significant influence of fetal sex on testosterone concentration with two non-overlapping distribution, one above and one below 300 pg/g fetal tissue, correlating with male and female gender respectively. After day 16, male fetuses tended to have higher testosterone concentrations, but some values obtained in females did overlap. Treatment of female mice with delta 9-tetrahydrocannabinol, the main psychoactive ingredient of marihuana, from days 12 to 16 pregnancy caused a significant (P less than 0.01) increase in fetal deaths in utero. Cannabinol treatment had no effect on this parameter, but reduced body weight (P less than 0.02) in female fetuses, and increased ano-genital distance (P less than 0.05) in male fetal mice. The concentrations of testosterone and dihydrotestosterone were reduced in male but not in female fetuses. The results indicate that exposure to psychoactive or non psychoactive constituents of marihuana suppresses testosterone levels in fetal as well as in immature and adult mice, as we have previously reported. Thus, maternal exposure to cannabinoids may interfere with the process of sexual differentiation in their male offspring as a result of decreased fetal androgen production. PMID- 6276488 TI - Effect of cortisol or adrenocorticotrophin on release of luteinizing hormone induced by luteinizing hormone releasing hormone in the dairy heifer. AB - During treatment with cortisol or ACTH, dairy heifers were given two doses of LH releasing hormone (LH-RH) spaced 1.5 h apart. Serum concentrations of cortisol and LH were monitored during each treatment. Treatment with both ACTH and cortisol raised plasma cortisol levels above the respective saline controls (P less than 0.001). Neither treatment affected basal LH concentrations. A slight depression in LH response was seen in the cortisol-treated animals after the first LH-RH injection, as shown by a statistically significant depression at three of the sample times. There was no significant difference between treated and control LH values after the second LH-RH administration. Treatment with ACTH resulted in significantly reduced LH values at all sample times after both injections of LH-RH. PMID- 6276489 TI - Responses of rat fetal adrenals to synthetic adrenocorticotrophic hormone and alpha-melanocyte-stimulating hormone in-vivo and in-vitro studies. AB - The response of the adrenals from rat fetuses at 16, 18 and 20 days of gestation to 1-24 ACTH and alpha-melanocyte-stimulating hormone (alpha-MSH) was studied in vitro. The response to 1-24 ACTH increased as gestation progressed. By the end of fetal life, corticosterone release induced by ACTH from whole adrenals was greater than that observed with adrenal tissue from non-pregnant adult female rats. High doses of alpha-MSH also stimulated adrenal activity but the response to ACTH was always higher than that to alpha-MSH. The effect of 1-24 ACTH and alpha-MSH on fetal adrenal growth was also compared in vivo. The adrenal atrophy induced by fetal hypophysectomy on day 17 of gestation could be prevented by i.m. administration of 10 microgram 1-24 ACTH or alpha-MSH. However, the adrenal growth was greater in ACTH-treated fetuses than in alpha-MSH treated ones. Later in gestation, between days 19 and 20, 1-24 ACTH but not alpha-MSH was able to prevent atrophy induced by fetal hypophysectomy. These findings are discussed in relation to the literature on levels of ACTH and alpha-MSH in the plasma and pituitary glands of the rat throughout the last third of gestation. High levels of ACTH in the fetal circulation contrast sharply with very weak or undetectable concentrations of alpha-MSH. Since the present data suggest that both trophic and steroidogenic activities of ACTH were greater than those of alpha-MSH, it may be concluded that ACTH but not alpha-MSH plays a major physiological role during gestation in the regulation of both fetal adrenal growth and function in the rat. PMID- 6276490 TI - Prolactin and luteinizing hormone receptors in marsupial corpora lutea: relationship to control of luteal function. AB - Prolactin and LH receptor concentrations in tammar wallaby corpora lutea (CL) have been examined and related to the control of luteal function in this and other marsupial species. During embryonic diapause, quiescent CL contained high concentrations of prolactin receptors. This was consistent with an earlier suggestion that prolactin may act directly on the CL to maintain its quiescent state. However, despite an apparent seasonal change in the mechanism by which the CL is maintained in quiescence, the number of luteal prolactin receptors remained constant throughout the year. Reactivation of quiescent CL led to an approximate halving of prolactin receptors on a per cell basis, but pregnancy had no effect. None of the wallaby CL showed any significant degree of LH binding, although wallaby follicles and testicular tissue did show binding. This lack of LH binding was consistent with previous findings that quiescent wallaby CL reactivate in response to hypophysectomy and are refractory to LH when cultured in vitro. Parturition, which occurs approximately 2 days before ovulation in this species, had no effect on numbers of either prolactin or LH receptors despite the occurrence of a peak of plasma prolactin at this time. The red kangaroo, which also exhibits embryonic diapause, showed a similar pattern of luteal receptor numbers with high prolactin and negligible LH binding, whereas the brush-tailed possum, which does not exhibit embryonic diapause, gave a pattern more like that seen in Eutheria with many more LH than prolactin receptors. PMID- 6276491 TI - Ocular disease pattern induced by herpes simplex virus is genetically determined by a specific region of viral DNA. AB - The pattern of ocular disease produced in the rabbit eye by HSV-1 (F) and HSV 1(MP) strains and recombinants F(MP)A, F(MP)B, F(MP)C, F(MP)D, F(MP)E, and F(MP)F was studied. The characteristics of ocular herpetic disease such as morphology of dendritic ulcers, severity of epithelial disease and incidence and duration of stromal disease produced in the rabbit eye are genetically determined by the virus strain. Our studies show that transfer of a defined part of the genome of the stromal disease-producing virus, HSV-1(MP), to the genome of an epithelial disease-producing virus, HSV-1(F), yielded recombinants with one or more of the disease characteristics of the donor strain. Specifically, recombinant F(MP)D produced lesions characteristic of the donor HSV-1(MP) strain; recombinants F(MP)C and F(MP)E produced stromal disease approaching the severity of the disease produced by the donor HSV-1(MP) strain, and only recombinants F(MP)A and F(MP)B retained the typically elongate lesions of the recipient HSV-1(F), whereas the recombinant strain F(MP)F produced no disease. The viral functions pertaining to the ocular disease pattern map between 0.70 and 0.83 map units in HSV-1 DNA within the BglII F DNA fragment. The pattern of stromal disease is independent of the production of glycoprotein C and fusion of HEp-2-infected cells. The functions relating to these aspects of ocular disease segregate but are closely linked. PMID- 6276492 TI - Genetic mapping of ecotropic murine leukemia virus-inducing loci in six inbred strains. AB - Mendelian segregation analysis was used to map chromosomal genes for the induction of endogenous N- and B-tropic ecotropic retroviruses (V loci) in high and low leukemic mouse strains. Patterns of virus expression were determined for mice of various inbred strains, congenic lines carrying single V loci, and the linkage testing stocks used in mapping studies. Segregation analysis resulted in the genetic mapping of V loci from six inbred strains to five mouse chromosomes. The V locus of A/J was mapped to chromosome 5 and shown to be allelic with that of BALB/cJ and C3H/HeJ (Cv); this suggests that Cv-represents a stable ancestral V locus present in Bagg albino stocks before the separation of inbred lines. The single, poorly inducible V locus of C57BL/10J and one of the four high virus loci of C58/Lw were mapped to the same region of chromosome 8 and may represent an allelic pair with different patterns of expression. An N-tropic V locus of the SEA/GnJ mouse was mapped to chromosome 9, and one of the three V loci of C3H/FgLw was mapped to chromosome 7. The endogenous B-tropic virus of B10.BR/SgLi was mapped to chromosome 11. These studies provide further evidence that endogenous ecotropic V loci are present at different chromosomal sites in unrelated mouse strains and emphasize the role of germ line reinfections in the generation of this diversity. PMID- 6276494 TI - Binocular summation occurs during interocular suppression. PMID- 6276493 TI - Role of the H-2 complex in induction of T helper cells in vivo. III. Contribution of the I-E subregion to restriction sites recognized by I-A/E-restricted T cells. AB - Previous studies on negative selection of T cells to sheep erythrocytes in irradiated mice showed that CBA (I-Ak,I-Ek) (kk) T cells comprise two subgroups of cells restricted by I-A (A alpha-A beta) and I-A/E (E alpha-E beta) molecules. Selection of the I-A/E-restricted by I-A (A alpha-A beta) and I-A/E (E alpha-E beta) molecules. Selection of the I-A/E-restricted subset requires that the donor T cells and the selection host share both I-A (E beta) and I-E (E alpha) gene products; only the I-A-restricted cells undergo selection in B10.A(4R) (kb) mice. This paper demonstrates that negative selection of the I-A/E-restricted subgroup of CBA T cells can occur in F1 hybrids between B10.A(4R) and various Ia.7+ (E alpha+) I-E-incompatible strains; selection does not occur in hybrids between B10.A(4R) and Ia.7- (E alpha-) strains. These data suggest that, despite the fact that E alpha chains display detectable structural allelic variations, these chains are functionally nonpolymorphic. This conclusion applies to E alpha k,d,p,r,j chains. With F1 hybrids between B10.A(4R) and another Ia.7+ strain, B10.PL (H-2u), in contrast, only intermediate selection is observed. This finding is consistent with recent evidence that cell surface expression of E alpha-u-E beta dimers displays strong cis preference. In contrast to E alpha+ CBA T cells, E alpha- B10.A(4R) (kb) T cells undergo complete negative selection in hosts matched only in the I-A (and H-2K) subregion, i.e., B10.BR (kk) mice; no selection occurs in B10 (bb) mice. These data imply that Ia-restricted T cells in E alpha- strains are probably restricted solely by I-A molecules. PMID- 6276495 TI - Stoichiometry of a half-turnover of band 3, the chloride transport protein of human erythrocytes. AB - The kinetics of human red blood cell Cl transport have been studied under nonequilibrium conditions to determine whether or not an outward Cl gradient can recruit the transport protein from an inward-facing to an outward-facing configuration. Three kinds of evidence are consistent with this outward recruitment. First, the initial net Cl efflux into a Cl-free phosphate medium is independent of the intracellular Cl concentration in the range 20-170 mM. Second, an outward Cl gradient strongly enhances the inhibitory potency of DNDS (4,4' dinitro-2,2'-stilbene disulfonate), which suggests that DNDS binds primarily to outward-facing states. Finally, we have estimated the number of Cl ions transported during the putative outward recruitment. Resealed red cell ghosts containing only 70 muM 36Cl were resuspended at 0 degrees C in a Cl-free, HCO3 free Na2SO4 medium. In the first 10 s, or approximately 10(6) Cl ions per ghost, followed by a much slower further loss of Cl. The rapid loss of 10(6) Cl ions per ghost, which is abolished by pretreatment with DIDS (4,4'-diisothiocyano-2,2' stilbene disulfonate), appears to represent the Cl that is transported during the first half-turnover of the transport cycle. These data are strong evidence that the influx and efflux events in the catalytic cycle for anion transport do not take place simultaneously, and that the stoichiometry of the transport cycle is close to one pair of anions exchanged per band 3 monomer. PMID- 6276496 TI - Titration of transport and modifier sites in the red cell anion transport system. AB - This work demonstrates the existence of titratable transport and modifier sites in the anion transport system of human red cells. Effects of alkaline extracellular pH on chloride exchange were studied up to pH 13 at 0 degrees C. The studies revealed two sets of reversible titratable groups. One set, having a pK of or approximately 11, appeared to be identical with the inhibitory halide binding modifier site. Deprotonation of this site stimulated anion transport. The apparent dissociation constants of chloride and iodide at this modifier site were 0.3 and 0.06 M, respectively, and it was confirmed that the organic sulfonate NAP taurine inhibits anion transport reversibly by a high-affinity interaction with halide-binding modifier sites at the extracellular side of the membrane. Other groups, with apparent pK of or approximately 12 at chloride concentrations above 0.1 M, were named as "transport sites" because transport function depended totally on their protonation. The apparent pK decreased when extracellular halide concentrations was lowered below 0.1 M. It was dependent of the intracellular chloride concentration, and was equally sensitive to extracellular pH of 13, was fully reversible. Hydroxyl ions were not transported to an appreciable extent by the anion exchange system. The pK values of both sets of groups make it likely that they are both arginyl residues, functioning as anion recognition sites similar to the role of functionally essential arginyl residues observed with numerous enzymes. PMID- 6276498 TI - The interrelation transketolase and dihydroxyacetone synthase activities in the methylotrophic yeast Candida boidinii. AB - Crude extracts of Candida boidinii grown on glucose, xylose or ethanol gave single peaks of classical transketolase activity following chromatography, on columns of hydroxylapatite; the enzyme was heat-stable and showed no appreciable activity with formaldehyde as acceptor in place of ribose 5-phosphate. Extracts of methanol-grown cells showed two peaks of transketolase activity following chromatography on both hydroxylapatite and DEAE-cellulose. One peak was identified with that found for the cells grown on substrates other than methanol; the other peak showed dihydroxyacetone synthase activity in addition to transketolase activity. Both activities in the latter peak were very unstable and have been ascribed to one enzyme on the basis of identical rates of denaturation at all temperatures tested between 0 and 40 degrees C. It is suggested that this enzyme is a special transketolase synthesized only during methylotrophic growth of the yeast and in contrast to classical transketolase, is capable of using equally well either formaldehyde or ribose 5-phosphate as glycolaldehyde acceptor. A method based on heat treatment has been suggested for the simultaneous assay of both transketolases present in crude extracts of a methylotrophically grown yeast. PMID- 6276497 TI - Irreversible inactivation of red cell chloride exchange with phenylglyoxal, and arginine-specific reagent. AB - Chloride exchange in resealed human erythrocyte ghosts can be irreversibly inhibited with phenylglyoxal, a reagent specific for the modification of arginyl residues in proteins. Phenylglyoxal inhibits anion transport in two distinct ways. At 0 degrees C, inhibition is instantaneous and fully reversible, whereas at higher temperature in an alkaline extracellular medium, covalent binding of phenylglyoxal leads to an irreversible inhibition of the transport membranes system. Indiscriminate modification of membrane arginyl residues was prevented by reacting the with phenylglyoxal in an alkaline extracellular medium while maintaining intracellular pH near neutrality. The rate of modification of anion transport depends on phenylglyoxal concentration, pH, temperature, and the presence of anions and reversible inhibitors of the anion transport system in fashions that are fully compatible with the conclusion that phenylglyoxal modifies arginyl residues that are essential for anion binding and translocation. Phenylglyoxal reacts rapidly with the deprotonated form of the reactive groups. It is proposed that the effects of anions and of negatively charged transport inhibitors on the rate of irreversible binding of phenylglyoxal are related to the effects of the anions on a positive interfacial potential. This potential determines the local pH, and thereby the concentration of deprotonated groups, in an exofacial region of the anion transport protein. PMID- 6276499 TI - Comparison of three methods for the purification of the delta haemolysin of Staphylococcus aureus. AB - Three methods for the purification of Staphylococcus aureus delta haemolysin were compared (Kantor et al., 1972; Kreger et al., 1971; Heatley, 1971). The products of these purifications from the culture supernatant of S. aureus strain RN25 were compared by electrophoresis, amino acid analysis, amino-terminal sequence analysis and thin-layer chromatography. The method of Heatley (1971) was found to be superior in terms of recovery and purity of the product. Delta haemolysin prepared by the method of Kreger et al. (1971) could not be sequenced successfully prior to treatment aimed at the removal of the N-formyl group at the amino-terminus. Delta haemolysin appears to exist in two distinct molecular forms, one with N-formylmethionine and the other with un-formylated methionine in the amino-terminal position. The former polypeptide species is purified preferentially by the method of Kreger et al. (1971). Thin-layer chromatography of the products of each method of purification revealed that they were all heterogeneous, although the major component from the product of the method of Heatley (1971) represented not less than 70% of the product. PMID- 6276500 TI - Rapid methods for the study of both stable and unstable plasmids in Pseudomonas. AB - Rapid methods for the analysis of degradative plasmids in Pseudomonas are described. Bacterial lysates prepared with alkaline sodium dodecyl sulphate were vigorously stirred in the presence of antifoam agent. Samples were subjected to agarose gel electrophoresis to detect plasmid DNA. After a short period of centrifugation on a sucrose gradient, the lysates yielded plasmid DNA of sufficient purity for restriction endonuclease digestion without further treatment. The methods have proved most useful in the extraction of previously undetected plasmid DNA, some of which appears to be unstable. PMID- 6276501 TI - [Oculo-dental-digital dysplasia: report of a case with spastic paraplegia]. PMID- 6276502 TI - C-reactive protein in acute viral infections. AB - A sensitive solid-phase enzyme immunoassay procedure was used to determine the concentrations of C-reactive protein (CPR) in the acute and convalescent phase sera of patients with verified rubella, herpes simplex, cytomegalo, influenza A or B, enterovirus, or mycoplasma infection. In all infection groups about 90% (80% for influenza) elevated CRP values were observed in the acute phase sera (mean values in the different groups 16-57 micrograms/ml), the highest values exceeding or approaching 100 micrograms/ml. The serum CRP values were highest in all groups before the specific serum antibodies were detectable and decreased approaching the upper limit or normal controls (2 microgram/ml) within 2 weeks. Notable individual variation in the CRP production was seen. We conclude tha serum CRP determination should not be used as a reliable criterion to distinguish bacterial and viral infections. PMID- 6276503 TI - Occurrence of antibodies against herpes simplex virus thymidine kinase in human sera. AB - The occurrence of antibodies in human serum that block herpes simplex virus (HSV) type 1 and 2 deoxythymidine kinase (dTK) has been investigated. The antibodies were detected by means of a sensitive assay technique using [125I]iododeoxyuridine as a substrate [Gronowitz and Kallander, 1980]. A total of 213 sera was studied. They included pairs of serum from patients with an acute HSV infection, individual sera from persons not suspected of a HSV infection, as well as sera from patients from whom HSV had been isolated. The HSV complement fixing (cf) titer was determined for each serum and subsequently used as a reference. None of the HSV cf negative sera contained HSV dTK blocking antibodies, whereas all cf positive sera did, however, all excepting those that were collected in connection with primary infections. by following the serum titer after a primary infection, we have found that 12 out of 19 persons studied had detectable dTK antibodies after an average time of 179 days. The results indicate a low degree of cross-reactivity between HSV type 1 and 2 dTK blocking antibodies. PMID- 6276504 TI - Development of serum and intestinal antibody response to rotavirus after naturally acquired rotavirus infection in man. AB - The temporal characteristics of the response of rotavirus specific IgM, IgG, IgA in serum and secretory antibody in feces to rotavirus were studied in 77 hospitalized patients with rotavirus induced gastroenteritis. The response in serum was characterized by the sequential appearance of rotavirus specific IgM, IgG, and IgA antibody. The IgM antibody appeared to be higher in the acute phase of the disease and was subsequently replaced by the IgG and IgA antibodies. However, the titers of IgG rotavirus antibody in convalescent specimens of serum were found to be statistically significantly lower in patients with severe or prolonged rotavirus infection than in specimens from subjects with mild or moderate disease. Most fecal specimens collected during both the acute and convalescent phase of illness contained virus specific secretory IgA. Higher concentrations of antibody were measured in convalescent samples from patients with prolonged diarrhea and virus shedding. These observations suggest a possible relationship between the severity of rotavirus infection and the nature of systemic and secretory antibody response. PMID- 6276505 TI - beta-Endorphin immunoreactivity and acute behavioral distress in children with leukemia. AB - Endogenous opiates have been implicated in pain and stress experiences. In order to directly assess the relationship between endorphin activity and acute behavioral distress, beta-endorphin immunoreactivity (beta-EPI) was measured by radioimmunoassay in cerebrospinal fluid of 75 children with acute leukemia undergoing routine lumbar puncture. These data were related to four measures of behavioral distress collected during the procedure. For children 4 years of age and above, beta-EPI correlated inversely with age (r = -.31,p less than or equal to .05). All behavioral measures also inversely correlated with age (r = -.26 to .67,p less than or equal to .05 to .001). Females had a significantly lower mean beta-EPI than males (p less than or equal to .01), and exhibited greater behavioral distress. beta-EPI and behavioral measures interacted with the use of specific antileukemia agents. L-Asparaginase was associated with lower beta-EPI (p less than or equal to .05), while prednisone was associated with lower behavioral distress on three of the four measures (p less than or equal to .05 to .01). After controlling for age, sex, and chemotherapy, beta-EPI and nurse ratings of anxiety were positively correlated (partial correlation coefficient = .31, p less than or equal to .05). Correlations between beta-EPI and other behavioral measures demonstrated positive trends. Results of this study are interpreted as support for the reactive nature of beta-EPI in cerebrospinal fluid to acute distress, and may help explain documented sex differences in distress behavior. Potential clinical implications and directions for further research are discussed. PMID- 6276507 TI - Alteration of 5'-nucleotidase and Na+,K+-ATPase in central and peripheral nervous tissue from dysmyelinating mutants (jimpy, quaking, trembler, shiverer, and mld). Comparison with CNPase in the developing sciatic nerve from trembler. PMID- 6276506 TI - Evidence that inhibition of nicotine-mediated catecholamine secretion from adrenal chromaffin cells by enkephalin, beta-endorphin, dynorphin (1-13), and opiates is not mediated via specific opiate receptors. AB - The opioid peptides Met- and Leu-enkephalin, dynorphin (1-13), and beta-endorphin and the narcotic analgesics, morphine, levorphanol, and dextrorphan all produced a dose-dependent inhibition of nicotine (5 x 10(-6) M)-mediated release of [3H]norepinephrine ([3H]NE) from bovine adrenal chromaffin cells in culture. None of these agents affected [3H]NE release induced by high K+ (56 mM). Although the above results suggest that the opioid peptides and narcotic analgesics inhibit catecholamine release from adrenal chromaffin cells in culture, we suggest that these effects are not mediated by specific opiate binding sites, since (1) the inhibition was only produced with high concentrations of the agents--the threshold concentrations were 10(-7) to 10(-5)M and higher; (2) the inhibition produced by the narcotic analgesics did not display stereospecificity, because the d-isomer, dextrorphan, was slightly more active than the l-isomer, levorphanol; (3) the narcotic antagonists naloxone, naltrexone, and levallorphan did not reverse the inhibition produced by either the narcotic analgesics (e.g., morphine) or the opioid peptides (e.g., dynorphin). These three antagonists themselves inhibited the nicotine-mediated release of [3H]NE from the adrenal chromaffin cells in culture. Finally (4), the I2-Tyr1 substituted analogues of beta-endorphin and dynorphin that are biologically less active than the parent compounds produced an inhibition of the nicotine-mediated [3H]NE release similar to that of their parent compounds. These results do not support the idea that high-affinity stereospecific opiate binding sites are involved in the inhibitory modulation of nicotinic evoked catecholamine release from bovine adrenal chromaffin cells in culture. PMID- 6276508 TI - Alterations of nuclear thyroid hormone receptors in cerebral cortex in vivo. AB - Investigation was conducted under in vivo conditions in the adult male rat to determine the basic characteristics of the nuclear thyroid hormone receptors in the cerebral cortex. Equilibrium with cortical nuclei of an intravenous dose of triiodothyronine (T3) occurred 3 h after injection and showed a t1/2 of 1 h for dissociation. Saturation of receptors occurred at 0.5--06. ng/mg DNA. The endogenous level of binding in the normal rat was 0.07--0.1 ng/mg DNA, representing a 15% occupancy of total receptors at a serum concentration of 66 ng/dl. These characteristics were then examined under several pathophysiological conditions. In the hypothyroid rat, an apparent 37% increase in total binding sites occurred. Under either fasting conditions or insulin or glucagon administration declines in serum T3 were noted, and the endogenous binding also decreased in parallel. Only glucagon produced a significant reduction in total binding sites. Under the hypoxic condition produced by maintenance under a 7% oxygen atmosphere, a slight increase in apparent total binding sites was found with no change in endogenous binding level. Severe narcosis resulted in no effects on T3 binding parameters. These results demonstrate specific alterations of thyroid hormone receptors that may be important physiologically. PMID- 6276509 TI - Glycine receptors in the human substantia nigra as defined by [3H]strychnine binding. AB - Specific [3H]strychnine binding was used to identify the glycine receptor macromolecular complex in human spinal cord, substantia nigra, inferior olivary nucleus, and cerebral cortex. In material from control patients a high-affinity KD (3--8 nM) was observed in the spinal cord and the substantia nigra, both the pars compacta and the pars reticulata. This is very similar to the values observed in the rat and bovine spinal cord (8 and 3 nM, respectively) and rat substantia nigra (12 nM). In the human brain the distribution of [3H]strychnine binding (at 10 nM) was: spinal cord = substantia nigra, pars compacta greater than substantia nigra, pars reticulata = inferior olivary nucleus greater than cerebral cortex. The binding capacity (Bmax) of the rat brain (substantia nigra or spinal cord) was approximately 10-fold that of the human brain. [3H]Strychnine binding was significantly decreased in the substantia nigra from Parkinson's disease patients, both in the pars compacta (67% of control) and the pars reticulata (50% of control), but not in the inferior olivary nucleus. The results were reproduced in preliminary experiment in rats with unilateral 6 hydroxydopamine lesions of the medial forebrain bundle. In the substantia nigra from patients who died with Huntington's disease, [3H]strychnine binding tended to be high (150% of control, NS) in both the pars compacta and the reticulata. [3H]Strychnine binding was unaltered in the substantia nigra of patients with senile dementia. Together with previous neurophysiological and neuropharmacological findings, those results support the hypothesis of glycine receptors occurring on dopamine cell bodies and/or dendrites in the substantia nigra. PMID- 6276510 TI - On the use of microwave radiation energy for brain tissue fixation. PMID- 6276511 TI - The distribution of enzymes which synthesize nicotinamide adenine dinucleotide and nicotinamide adenine dinucleotide phosphate in monkey, rabbit, and ground squirrel retinas. AB - The activity of adenosinetriphosphate:nicotinamide adenylyltransferase (EC 2.7.7.1) was measured in all the layers of monkey, rabbit, and ground squirrel retinas. Nicotinamide adenine dinucleotide (NAD) kinase (EC 2.7.1.23) distribution was measured in monkey and rabbit retinas. An attempt was made to measure NAD synthetase (EC 6.3.5.1), but the activities in the retinal layers were too low to produce a reliable increment in the levels of endogenous NAD. In monkey retina the adenylyl transferase was highest by far in the outer and inner nuclear layers, lower and variable in ganglion cell and fiber layers, and almost absent elsewhere. Rabbit retina differed in that activity was nearly absent in the outer nuclear layer, whereas in the ground squirrel outer nuclear layer activity was double that of the inner nuclear layer. The species differences suggest that adenylyl transferase is almost absent from cone cell nuclei and high in rod cell nuclei. NAD kinase distribution in monkey retina was almost the mirror image of that of adenylyl transferase. PMID- 6276512 TI - Muscle uptake of 99mtechnetium pyrophosphate in patients with neuromuscular disorder. A quantitative study. AB - 99mTechnetium pyrophosphate (99mTc PYP) muscular scanning was carried out in 28 patients affected by neuromuscular diseases. To express the muscle tracer retention quantitatively, the soft tissue to bone uptake ratio was calculated in the thigh and in the lower leg. Significantly increased ratio values were found in all the patients with polymyositis and in most patients with different types of muscular dystrophy, while high values were seen in a few subjects with myasthenia gravis and with neurogenic disorders. These findings are compared with those of previous studies, and the factors playing a role in determining some discrepant observations are discussed. The use of 99mTc muscular scanning in the investigation of patients with suspected inflammatory myopathy is suggested. PMID- 6276513 TI - A case of adrenoleukodystrophy in a girl. Genetic considerations. AB - We report the case of a girl who developed leukoencephalopathy and adrenal atrophy and died at 3 years of age. Histologically, demyelination, gliosis, perivascular lymphocytic cuffing and sudanophilia were present in the brain. The adrenal cortex was atrophic. Ultrastructurally, there were numerous cytoplasmic inclusions in brain macrophages, consisting of two leaflets separated by an intervening space of variable low electron density. Brain tissue cholesterol esters contained a high proportion of long chain fatty acids. The findings are discussed and compared with those in the literature. Emphasis is placed on the fact that the disease occurred in a girl in apparent contradiction to the commonly accepted X-linked hereditary transmission of adrenoleukodystrophy. Some possible genetic explanations for our case are put forward. PMID- 6276514 TI - Familial progressive myoclonic epilepsy. A clinical, genetical, biochemical and patho-anatomical study of a family with a 6-year follow-up. AB - Six siblings, including 4 cases of myoclonic epilepsy, their parents and 2 grandmothers were subjected to systematic investigation, and the patients were followed-up. The genetic studies revealed in the mother's family a patient with Lafora bodies demonstrated at autopsy. No chromosome abnormalities were found nor any linkage to the HLA system. The affected family members were characterized biochemically by an increased excretion of total glycosaminoglycans and/or an abnormal electrophoretic pattern of urinary glycosaminoglycans with an increased proportion of low-sulfated glycosaminoglycans. In the healthy family members this pattern of electrophoresis could also be demonstrated in the father and the paternal grandmother. Based on the biochemical results and the genetic studies it is suggested that the family members with progressive familial myoclonic epilepsy present a combination of at least 2 hereditary defects. The course of the disease has been relatively benign and treatment with sodium valproate, baclofen and clonazepam has shown quite satisfying results. In consequence of the biochemical findings combined treatment with A and E vitamins has been initiated. PMID- 6276515 TI - Chronic removal of inhibitory axon alters excitatory transmission in a crustacean muscle fiber. PMID- 6276516 TI - Non chromaffin paragangliomas of the head and neck. Diagnostic and therapeutic angiography in 19 cases explored from 1977 to 1980. PMID- 6276517 TI - Embolization of the cavernous and petrous segments of the internal carotid artery in severe basilar skull and petrous bone lesions. PMID- 6276518 TI - Vitamin B-6 metabolism and its relation to ornithine decarboxylase activity in regenerating rat liver. AB - The metabolism of vitamin B-6 regenerating rat liver and liver from sham-operated control animals fed either a pyridoxine-sufficient or pyridoxine-deficient was investigated. The pyridoxal phosphate levels in plasma, regenerating liver and control liver were determined as were the activities of three enzymes involved in the metabolism of the vitamin, namely, pyridoxine kinase, pyridoxine phosphate oxidase, and pyridoxine phosphate phosphatase. In addition, total and holo ornithine decarboxylase activities in the livers were measured. The results indicate that the metabolism of vitamin B-6 regenerating rat liver is different from that observed in Morris hepatomas (Thanassi et al. (1981) J. Biol. Chem. 256, 3370-3375). Vitamin B-6 metabolism in Morris hepatomas is concluded to be characteristic of the tumors rather than a property common to rapidly proliferating hepatic tissue. Regenerating liver ornithine decarboxylase holoenzyme activity in pyridoxine deprived rats was maintained at the same level as that in regenerating liver of pyridoxine-sufficient animals. The mechanism behind this maintenance of holo-enzyme activity appears to involve a pronounced increase in the amount of apoornithine decarboxylase. The time-dependent peak of ornithine decarboxylase activity following partial hepatectomy was shifted from four hours to twelve hours by vitamin B-6 deficiency. PMID- 6276519 TI - Effects on Ca and P metabolism in humans by adding meat, meat plus milk, or purified proteins plus Ca and P to a low protein diet. AB - The effects on calcium and phosphorus metabolism of adult man by adding meat or meat plus dairy products to a diet low in protein (55 g), calcium (590 mg), and phosphorus (890 mg) were determined. When the low protein diet was consumed, the subjects retained a mean of 20 mg calcium daily but lost 106 mg phosphorus. The addition of meat which increased protein and phosphorus to 146 g and 1660 mg, respectively, caused calcium retention to decrease from 19 to -17 mg but phosphorus retention to increase from 106 to 55 mg. When the meat plus dairy diet high in protein (146 g), calcium (1370 mg), and phosphorus (2060 mg) was consumed the subjects retained substantial amounts of calcium (101 mg) and phosphorus (177 mg). The simulated diets high in purified proteins and supplemented with calcium and phosphorus affected calcium retention in a manner similar to the meat and meat plus dairy diets, but they had a marked negative effect on phosphorus retention; this indicates that supplements of calcium gluconate were well utilized but that those of monopotassium phosphate were not. The results obtained on urinary sulfate, acid, cyclic AMP and hydroxyproline support the conclusions made from the calcium and phosphorus data. PMID- 6276520 TI - Effects of several detergents and dietary fiber added to a diet on intestinal sucrase activity in rats. PMID- 6276521 TI - Virus-associated hemophagocytic syndrome or malignant histiocytosis? PMID- 6276522 TI - The value of neuroradiology in infantile spasms. AB - The results of neuroradiologic studies of 71 children with infantile spasms treated with ACTH were correlated with the developmental outcome at follow-up (mean 62 months, range 14 to 207 months). Fifty-two (73%) patients had an abnormal NRS on initial evaluation; 49% had cerebral atrophy, 18% had congenital anomalies, and 6% had hydrocephalus. Twenty patients were normal on initial clinical evaluation. At follow-up only the eight (40%) with normal NRS were normal. Twelve (60%) who had unexpected abnormalities on NRS were retarded at follow-up. CAT scanning is necessary to predict the developmental outcome in developmentally normal children with infantile spasm. Eight of nine patients with normal NRS in the early treatment group were developmentally normal at presentation and follow-up. Ten patients who were developmentally normal before spasms began, and had normal NRS but were in the late treatment group, were retarded at initial evaluation and follow-up. This finding suggests that early treatment of children who have a normal NRS and normal development at onset of spasm prevents mental retardation. PMID- 6276524 TI - Effects of PEEP and tolazoline infusion on respiratory and inert gas exchange in experimental meconium aspiration. AB - Meconium aspiration syndrome often produces respiratory failure in the neonate. We utilized the multiple inert gas elimination technique to study the effects on respiratory and inert gas exchange of the application of positive end expiratory pressure or continuous infusion of tolazoline HCl. The application of PEEP, with the optimal level of PEEP defined for each animal, produced a decrease in AaDO2 and pulmonary shunt, without an increase in blood flow to low VA/Q areas, or an increase in dead space. Tolazoline infusion, at 2 mg/kg/hour, had no apparent effect on AaDO2 or shunt, or magnitude of low VA/Q regions. Tolazoline therapy was associated with an increase in heart rate and a decrease in systemic blood pressure. We conclude that immediate postaspiration application of PEEP, but not of tolazoline, will diminish pulmonary shunt without creating low VA/Q areas, and therefore will improve gas exchange in MAS. PMID- 6276523 TI - Liver malignancy after parenteral nutrition. PMID- 6276527 TI - Interstitial conduction and the emergent mind. PMID- 6276525 TI - The effect of silica injections on the rejection of Eimeria from nonspecific hosts. PMID- 6276526 TI - Immunological response to herpes simplex virus in human gingival fluid. PMID- 6276528 TI - Effect of 4-dimethylaminomethyl-1-(3-hydroxyphenyl)-1-nonen-3-one hydrochloride and related compounds on respiration in rat liver mitochondria. AB - 4-Dimethylaminomethyl-1-(3-hydroxyphenyl)-1-nonen 3-one hydrochloride (II) was shown to stimulate respiration in rat liver mitochondria at levels of 2.5 mumoles or less; but at levels higher than 5.0 mumoles, respiration was inhibited when succinate and 3-hydroxybutyrate were the substrates. Compound II caused inhibition of respiration in the presence of glutamate over the dose range studied. The stimulating effect of II was attributed to its functioning as an uncoupling agent. Its inhibiting properties were considered to be due to its behaving like antimycin A in blocking transport of electrons between cytochromes b and c1. The effect of II on respiration in mitochondria varied with the pH of the medium. A conjugated styryl ketone, which contained a nuclear hydroxy function and was structurally related to II, also stimulated respiration at low doses while inhibiting respiration at higher concentrations. Etherification of the hydroxy group led to compounds in which only stimulation of respiration was noted. PMID- 6276529 TI - Quantitative evaluation of pharmaceutical effervescent systems I: Design of testing apparatus. AB - Two new devices were developed to monitor the reactivity of pharmaceutical effervescent systems. The first method monitored carbon dioxide pressure generation during the effervescent reaction in a plastic pressure vessel fitted with a pressure gauge. The second method monitored weight loss, attributed to carbon dioxide loss to the atmosphere, by means of a double cantilever beam and an electromagnetic proximity transducer. In the pressure device, the quantification of an effervescent reaction was accomplished by measuring the dissolution time of the effervescent system and the pressure generated. Quantification of an effervescent reaction using the beam device utilized the total carbon dioxide weight loss, the rate of weight loss, and the effervescent reaction lag time. PMID- 6276530 TI - Heterogeneity of biochemical actions among vasodilators. AB - Thirty-four vasodilators were screened in several in vitro biochemical assays related to smooth muscle excitation-contraction coupling, binding to beta 1-,beta 2-, and alpha-adrenergic receptors, inhibition of phosphodiesterase activity, and antagonism of calcium accumulation. Isoproterenol and perhexiline only exhibited binding to beta-adrenergic sites. Ergocryptine, tolazoline, and amotriphene only bound to alpha-adrenergic receptors. Leniquinsin, papaverine, proquazone, dioxyline, hoquizil, quazodine, and theophylline were active only as phosphodiesterase inhibitors. Isoxsuprine, nylidrin, and bencyclane bound to alpha- and beta-receptors. Pentoxifylline bound to beta 1-sites and inhibited phosphodiesterase. Cyclandelate bound to beta 2-sites and blocked calcium accumulation. Cinnarizine and flunarizine antagonized calcium accumulation and bound to alpha-sites. Prazosin bound to alpha-sites and inhibited phosphodiesterase. Ethaverine and dipyridamole were inhibitors of phosphodiesterase and calcium accumulation. Nafronyl bound to beta 2- and alpha sites and antagonized calcium accumulation. Mebeverine bound to beta 2- and alpha receptors and inhibited phosphodiesterase activity and calcium accumulation. Verapamil bound to alpha-sites, and blocked phosphodiesterase and calcium accumulation. Quinazosin bound to beta 2- and alpha-receptors and antagonized both phosphodiesterase activity and calcium accumulation. Vasodilators that were inactive in all assays included niacin, nicotinyl alcohol, inositol nicotinate, amyl nitrite, sodium nitroprusside, diazoxide, hydralazine, and protoveratrine. Vasodilators should not be considered as a single drug class since they act on various mechanisms related to coupling of neuronal excitation to muscular contractility. PMID- 6276531 TI - Protein binding of N-2-mercaptoethyl-1,3-diaminopropane via mixed disulfide formation after oral administration of WR 2721. AB - Earlier studies have shown that WR 2721 [H2N-(CH2)3-NH(CH2)2SPO3H2] is converted to its free thiol form, N-2-mercaptoethyl-1,3-diaminopropane (MDP), at the acidic pH of the stomach. MDP is a radioprotective compound and a mucolytic agent capable of decreasing sputum viscosity in the lungs of patients with cystic fibrosis. Conversion of WR 2721 and MDP to the corresponding sulfonic acid (MDP SO3H) permits quantitative determination of these compounds in physiological fluids by use of an automatic amino acid analyzer. After oral administration of WR 2721 to human patients and rabbits it is converted to MDP and the free thiol form of the drug associates with plasma proteins by mixed disulfide linkage. The plasma proteins serve as a depot and reservoir of MDP for potential exchange at the tissues. When incubated with whole sputum or with purified mucin solutions in vitro, MDP decreased the viscosity of these solutions by reduction of the accessible disulfide bonds of the mucin molecule and was subsequently found in mixed disulfide association with the mucin molecule. The association of MDP with proteins via mixed disulfide linkage has important implications for the development of optimal dose regimens for administration of WR 2721 to patients. PMID- 6276532 TI - The effect of acute opioid administration on the phosphorylation of rat striatal synaptic membrane proteins. AB - The acute administration of morphine, methadone, etorphine, methionine enkephalin, leucine-enkephalin and some related pharmacologically active peptides to rats produces biphasic effects on the degree of phosphorylation of striatal synaptic plasma membrane (SPM) proteins by endogenous protein kinases in the membranes. At 1 to 5 min after opioid administration, the phosphorylation of several SPM proteins is enhanced, followed at 10 to 20 min by a severe reduction in the rate of phosphorylation of the same proteins. Multiphasic responses follow the initial biphasic response after the injection of some opioids. Naloxone is able to block the agonist effects. The injection of saline or an inactive narcotic isomer has no effect on the phosphorylation of striatal SPM proteins. The administration of other psychoactive drugs such as clonidine or haloperidol also produces changes in phosphorylation, although the patterns of change are distinctly different from those produced by opioids. The proteins affected by opioid administration resemble those susceptible to stimulation of phosphorylation in the presence of Ca++/calmodulin. The role of Ca++/calmodulin regulated reactions in the mechanisms of action of opioids has been evaluated by the addition of these modulators to the assays of preparation from opioid-treated rats. Neither modulators, added separately or together to the assay, was able to restore a depressed phosphorylation to normal levels, although each modulator was effective in stimulating the phosphorylation of many SPM proteins. PMID- 6276533 TI - Aminoglycoside inhibition of a renal phosphatidylinositol phospholipase C. AB - Aminoglycosides are able to inhibit phosphatidylinositol phospholipase C activity in a soluble fraction of rat renal tissue. The Km of the enzyme with respect to phosphatidylinositol is 0.53 mM and Vmax was 0.671 mumol of phosphatidylinositol cleaved per hr per mg of protein. The enzyme is calcium activated and inhibited by ethylene glycol bis-(beta-aminoethyl ether)-N,N'-tetraacetic acid. Streptomycin, amikacin, kanamycin, tobramycin, gentamicin and neomycin inhibit enzyme activity in the same rank order as their ability to produce nephrotoxicity. The 50% inhibitory concentrations ranged from 0.03 mM for neomycin to 0.38 mM for streptomycin. Lineweaver Burk plots indicate competitive inhibition with a possible relation to the calcium activation. It is possible that aminoglycosides may inhibit phosphatidylinositol phospholipase C in vivo and that this inhibition may be related to the nephrotoxicity of aminoglycosides. PMID- 6276534 TI - Specific inhibition of calcium channels by calcium ions in smooth muscle. PMID- 6276535 TI - Alteration in sensitivity to isoproterenol and acetylcholine in the rat heart after repeated administration of isoproterenol. AB - Alteration of sensitivity to the inotropic responses to isoproterenol and acetylcholine (ACh) and of saturation constants for the specific binding of [3H]dihydroalprenolol and [3H]quinuclidinyl benzilate to particulate fractions were investigated in rat hearts chronically treated with isoproterenol. The EC50 value for the inotropic response to isoproterenol in the atria of rats injected i.p. with isoproterenol (0.01 or 0.1 mg/kg) twice a day for 10 days, was 9.0-fold higher than in controls. Isoproterenol (0.17 mg/kg i.p.)-induced stimulation of cardiac ornithine decarboxylase activity was significantly (P less than .05) attenuated in atria from animals of the treated groups. Scatchard analysis of specific binding of [3H]dihydroalprenolol revealed that repeated isoproterenol injection produced a significant decrease in the maximum number of binding sites (Bmax), but not in the dissociation constant (KD) for this antagonist. In comparison to the isoproterenol-induced changes in beta adrenergic positive inotropic response, repeated treatment with isoproterenol decreased 2-fold the EC50 value for the negative inotropic response to ACh in isolated atria. Scatchard analysis of specific [3H]quinuclidinyl benzilate binding, however, indicated a significant (P less than .01) decrease in Bmax. Repeated isoproterenol injections also elicited a significant increase in cardiac weight. It is suggested that repeated administration of isoproterenol induces a decrease in density of beta adrenergic receptors which may in part underlie a cardiac hyposensitivity to isoproterenol and an increase in the number of muscarinic ACh receptors and inotropic responsiveness to ACh. PMID- 6276536 TI - The renin-angiotensin system in aminoglycoside-induced acute renal failure. AB - To examine the role of the renin-angiotensin system in aminoglycoside-induced acute renal failure, we gave rats gentamicin (80 mg/kg/day), gentamicin + captopril (an angiotensin-converting enzyme inhibitor), captopril alone or saline sham injections, for 10 days. Blood pressure, sodium excretion, urine osmolality and creatinine clearance were measured four times during treatment. Plasma renin activity, angiotensin I-converting enzyme and renal histology were determined at sacrifice. Although angiotensin I-converting enzyme activity was markedly suppressed by captopril treatment, the addition of captopril failed to consistently influence blood pressure or urine osmolality. In addition, it did not promote natriuresis in the face of gentamicin toxicity. Creatinine clearance decreased progressively in groups receiving gentamicin and was lowest (P less than .05) in the groups receiving gentamicin + captopril. Captopril did not prevent the development of tubular epithelial nor glomerular endothelial changes associated with gentamicin toxicity. In a separate experiment rats received captopril for 3 days prior, before receiving the gentamicin + captopril regimen for 10 days. Captopril pretreatment did not influence the results. This study does not support a pivotal role for the renin-angiotensin system in the production of the decreased glomerular filtration rate observed in nephrotoxic acute renal failure induced by gentamicin in rats. PMID- 6276537 TI - Characterization of ribonuclease A irradiated with gamma-rays in the presence of cytidylic acid with respect to the interaction of the enzyme with folic acid. PMID- 6276538 TI - Calcium pyrophosphate dihydrate deposition in lumbar disc fibrocartilage. AB - Calcium pyrophosphate dihydrate deposits were found in the lumbar disc fibrocartilage in 4 patients undergoing surgery for spinal cord or nerve root compression. All of the patients had prior surgery at the same lumbar area. None of the patients had the articular or roentgenographic manifestations of calcium pyrophosphate deposition disease (pseudogout). Andres and Trainer have recently reported 7 similar patients. Calcium pyrophosphate dihydrate deposition in axial skeleton fibrocartilage may be a common finding in patients undergoing repeat spinal surgery. PMID- 6276539 TI - Cartilage matrix in hereditary pyrophosphate arthropathy. AB - Morphological and biochemical studies of articular cartilage were performed in 6 members of 3 families with hereditary pyrophosphate arthropathy. Evidence of metabolic disturbance of cartilage matrix was obtained from light and electron microscopic findings and by the content and composition of glycosaminoglycans. PMID- 6276540 TI - Evidence of the preferential involvement of mu receptors in analgesia using enkephalins highly selective for peripheral mu or delta receptors. AB - In order to study the preferential involvement of mu or delta receptors in the analgesic effects of enkephalins, several peptides which selectively interact with these two kinds of receptors in peripheral organs were synthesized. The inhibitory potency on the electrically stimulated mouse vas deferens (delta receptors) of the short peptide Tyr-D-Ala-Gly-NH-CH(CH3)CH2CH(CH3)2 (6) is 2100 times lower (IC50 = 1220 nM) than that of the longer and more hydrophilic peptide Tyr-D-Ser-Gly-Phe-Leu-Thr (10) (IC50 = 0.58 nM). In contrast, the IC50 values of all the synthesized compounds on the guinea pig ileum assay (mu receptors) are in the same range (100-360 nM). Likewise, their analgesic activities in mice, measured on the hot-plate test after intracerebroventricular injection, are similar. Therefore, the dissociation between antinociceptive properties in mice and potencies on the mouse vas deferens unambiguously reflects a preferential implication of mu receptors in analgesia. The possible involvement of brain delta receptors in behavioral effects is discussed. PMID- 6276541 TI - Synthesis and biological activity of certain derivatives of oxazinomycin and related oxadiazole nucleosides. AB - Oxazinomycin was converted into 2',3',5'-tri-O-acetyloxazinomycin (2) and 2',3'-O isopropylideneoxazinomycin (3), respectively. Compound 3 was iodinated and reduced to provide 5'-deoxy-2',3'-O-isopropylideneoxazinomycin (5) which, after acid hydrolysis, provided 5'-deoxyoxazinomycin (6). Alternatively, the iodination of oxazinomycin followed by catalytic hydrogenation also provided 6. Oxazinomycin was treated with 2-acetoxybenzoyl chloride to provide 3'-O-acetyl-2'-chloro-2' deoxyoxazinomycin (8) which, after reduction with tributyltin hydride, provided 3'-O-acetyl-2'-deoxyoxazinomycin (9). Oxazinomycin was also converted into oxazinomycin 5'-phosphate (10) and into O4,2'-anhydrooxazinomycin (11). 1,2,4 Oxadiazole-3,5-dione (12) was glycosylated to provide 2-(2,3,5-tri-O-acetyl-beta D-ribofuranosyl)-1,2,4-oxadiazole-3,5-dione (13) which, after deacetylation, provided 2-beta-D-ribofuranosyl-1,2,4-oxadiazole-3,5-dione (14). Similarly, 12 provided 2-(2-deoxy-beta-D-erythro-pentofuranosyl)-1,2,4-oxadiazole-3,5-dione (17); 14 was also converted into the corresponding 2',3'-O-isopropylidene derivative 15. Compound 14 showed significant antiviral activity against herpes simplex virus type 1, in vitro. PMID- 6276542 TI - Potential antiestrogens. Synthesis and evaluation of mammary tumor inhibiting activity of 1,2-dialkyl-1,2-bis(3'-hydroxyphenyl)ethanes. AB - The syntheses of the meso-1,2-dialkyl-1,2-bis(3'-hydroxyphenyl)ethanes [alkyl substituent: CH3 (19), C2H5 (20), C3H7 (22), C4H9 (23), i-C4H9 (24), and C5H11 (25)] and of d,l-3,4-bis(3'-hydroxyphenyl)hexane (21) are described. In vitro these compounds inhibited the [3H]estradiol receptor interaction competitively, exhibiting Ka values between 0.20 x 10(9) (20) and 0.11 x 10(6) M-1 (24). In vivo the meso compounds reduced the estrone-stimulated mouse uterine growth; the most effective compounds were 20, 22, and 23 (53, 50, and 45% inhibition, respectively). Compounds 20 and 22-24 showed weak estrogenic activity in the mouse uterine weight test and in the vaginal cornification test. Compounds 19 (NSC-297169), 20 (NSC-297170), and 22 (NSC-297171) exhibited a dose-dependent growth inhibition on the MCF-7 human breast tumor cell line (10(-6) to 10(-9) M). These compounds also showed a marked dose-dependent inhibition on the DMBA induced, hormone-dependent mammary carcinoma of the Sprague-Dawley rat corresponding to their association constants. PMID- 6276543 TI - Membrane effects of antiinflammatory agents. 1. Interaction of sulindac and its metabolites with phospholipid membrane, a magnetic resonance study. AB - High-resolution proton NMR and spin-label ESR spectroscopies have been applied to examine the interaction of the nonsteroidal antiinflammatory drug sulindac (1) and its active sulfide metabolite (2) and inactive sulfone metabolite (3) with phospholipid membranes. Only weak interactions were observed with 1 and 3, but a strong interaction with 2 was indicated both by specific changes in the proton transverse relaxation rate (1/T2) of different substituents in 2 and by a unique shift in membrane transition temperature in the presence of 2 as measured by the ESR technique. Since the structural differences of these compounds are confined to a single polar substituent, i.e., the oxidation state of the sulfur atom, the strong interaction of the sulfide metabolite (2) with the neutral phospholipid membrane is ascribed to its high partition coefficient in the lipid membrane and its ability to penetrate into the lipid bilayer with the carboxyl group remaining at the polar membrane surface. As evidenced from the ESR spectra of two spin labels, C5- and C12-doxylstearic acid, no significant change of the membrane fluidity was induced by the interaction of 2 with phospholipid vesicles. PMID- 6276544 TI - Inhibition of separated forms of cyclic nucleotide phosphodiesterase from pig coronary arteries by 1,3-disubstituted and 1,3,8-trisubstituted xanthines. AB - A series of xanthines with varied substituents in the 1, 3, and 8 positions were prepared in an attempt to understand the structure--activity relationship for alkylxanthines as inhibitors of two different forms of cyclic nucleotide phosphodiesterase. Polar substituents on the 1 or 3 position of the xanthine reduced the potency of the xanthines to inhibit both the calmodulin-sensitive and the "cyclic AMP specific" forms of phosphodiesterase. Polar substituents on the 8 position of the xanthine, other than a carboxylic acid, increased the potency to inhibit the calmodulin-sensitive form of phosphodiesterase, if they were capable of donating electrons to the xanthine nucleus. On the other hand, any substituent in the 8 position larger than H reduced the potency of the xanthines to inhibit the cyclic AMP specific form of phosphodiesterase. Topographical maps of the active sites of the two forms of phosphodiesterase are presented in summary. PMID- 6276545 TI - Aminomethyl-1,2,4-benzothiadiazines as potential analogues of gamma-aminobutyric acid. Unexpected discovery of a taurine antagonist. AB - A series of 6- and 8-(aminomethyl)-4H-1,2,4-benzothiadiazine 1,1-dioxides has been synthesized and tested for interaction with various GABA systems. None of the compounds showed significant GABA-mimetic properties, but unexpectedly, compound 7 [6-(aminomethyl)-3-methyl-4H-1,2,4-benzothiadiazine 1,1-dioxide] possessed the properties of a selective antagonist of taurine, as measured by the antagonism of taurine-induced inhibition of rat cerebellar Purkinje firing. PMID- 6276546 TI - Common anionic receptor site hypothesis: its relevance to the antagonist action of naloxone. AB - Appropriate modification of 14 beta-methoxy- and 14 beta-ethoxycodeinone (prepared by alkylation of 14 beta-hydroxycodeinone) has generated four alkoxy analogues (3a-d) of naloxone and naltrexone. These agents were pure narcotic antagonists in contradiction to the predictions of the common anionic receptor site hypothesis, postulated to be of importance in the enhanced antagonism of naloxone. The molecular change from allyl to cyclopropylmethyl on the N atom increased selectivity of these antagonists for the mu receptor to the same extent as found for naloxone. Increase in the length of the C14 O-substituent had no effect on receptor selectivity, and either formation in most cases did not significantly alter oral/parenteral ratios of durations of action. PMID- 6276547 TI - Single-file diffusion multi-ion mechanism of permeation in paracellular epithelial channels. AB - The transepithelial fluxes, conductances and permeabilities of Li+, Na+, K+, Cs+, NH4+ and H3CNH3+ were studied under ionic concentrations ranging from 12 to 250 mM in Bufo arenarum gallbladders. When these measurements are carefully corrected in order to get only the component due to the paracellular cation channels, the following results are obtained: (1) The permeability ratios (cationic/anionic) are a decreasing function of salt concentration. (2) The partial conductances through paracellular cationic channels show nonlinear saturable concentration kinetics. (3) Moreover, partial conductance kinetics of K+, Cs+ and NH4+ present a maximum followed, at higher concentrations, by a negative-slope region. (4) The selectivity sequences obtained from biionic potentials do not agree with those obtained from partial conductance measurements. (5) The unidirectional 22Na tracer flux (serosal to mucosal) is inhibited by 63% when the K+ symmetrical concentration in the bathing solutions is raised from 25 to 200 mM. (6) When the unidirectional 42K fluxes (serosal to mucosal) at 200 mM KCl Na-free solutions are compared with K+ partial conductance by means of the Hodgkin and Keynes (Hodgkin, A.L., Keynes, R.D. 1955, J. Physiol London 128:61-88) expression, the n' factor is 2.0. These results indicate that cations do not follow the independence principle and behave as in single-file diffusion multi-ion pores when crossing the paracellular cation channels of Bufo arenarum gallbladder epithelium. PMID- 6276548 TI - Effects of divalent cations on toad end-plate channels. AB - Miniature end-plate currents (MEPCs) and acetylcholine-induced current fluctuations were recorded in voltage-clamped, glycerol-treated toad sartorius muscle fibers in control solution and in solutions with added divalent cations. In isosmotic solutions containing 20 mM Ca or Mg, MEPCs had time constants of decay (tau D) which were about 30% slower than normal. In isotonic Ca solutions (Na-free), greater increases in both tau D and channel lifetime were seen; the null potential was -34 mV, and single-channel conductance decreased to approximately 5 pS. Zn or Ni, at concentrations of 0.1-5 mM, were much more effective in increasing tau D than Ca or Mg, although they did not greatly affect channel conductance. The normal temperature and voltage sensitivity of tau was not significantly altered by any of the added divalent cations. Surface potential shifts arising from screening of membrane fixed charge by divalent cations cannot entirely explain the observed increases in tau, especially when taken together with changes in channel conductance. PMID- 6276550 TI - Aldosterone control of the density of sodium channels in the toad urinary bladder. AB - Near-instantaneous current-voltage relationships and shot-noise analysis of amiloride-induced current fluctuations were used to estimate apical membrane permeability to Na (PNa), intraepithelial Na activity (Nac), single-channel Na currents (i) and the number of open (conducting) apical Na channels (N0), in the urinary bladder of the toad (Bufo marinus). To facilitate voltage-clamping of the apical membrane, the serosal plasma membranes were depolarized by substitution of a high KCl (85 mM) sucrose (50 mM) medium for the conventional Na-Ringer's solution on the serosal side. Aldosterone (5 X 10(-7) M, serosal side only) elicited proportionate increases in the Na-specific current (INa and in PNa, with no significant change in the dependence of PNa on mucosal Na (Nao). PNa and the control of PNa by aldosterone were substrate-dependent: In substrate-depleted bladders, pretreatment with aldosterone markedly augmented the response to pyruvate (7.5 X 10(-3) M) which evoked coordinate and equivalent increases in INa and PNa. The aldosterone-dependent increase in PNa was a result of an equivalent increase in the area density of conducting apical Na channels. The computed single-channel current did not change. We propose that, following aldosterone induced protein synthesis, there is a reversible metabolically-dependent recruitment of preexisting Na channels from a reservoir of electrically undetectable channels. The results do not exclude the possibility of a complementary induction of Na-channel synthesis. PMID- 6276549 TI - The role of sodium-channel density in the natriferic response of the toad urinary bladder to an antidiuretic hormone. AB - Urinary bladders of Bufo marinus were depolarized, by raising the serosal K concentration, to facilitate voltage-clamping of the apical membrane. Passive Na transport across the apical membrane was then studied with near-instantaneous current-voltage curves obtained before and after eliciting a natriferic response with oxytocin. Fitting with the constant-field equation showed that the natriferic effect is accounted for by an increase in the apical Na permeability. It is accompanied by a small increase in cellular Na activity. Furthermore, fluctuation analysis of the amiloride-induced shot-noise component of the short circuit current indicated that the permeability increase is not due to increased Na translocation through those Na channels which were already conducting prior to hormonal stimulation. Rather, the natriferic effects is found to be based on an increase in the population of transporting channels. It appears that, in response to the hormone, Na channels are rapidly "recruited" from a pool of electrically silent channels. PMID- 6276551 TI - Preparation and properties of spin-labeled lecithin-cholesterol liposomes. AB - Lecithin-cholesterol vesicles of various compositions containing membrane-bound spin-labeled cholestane can be prepared by appropriate choice of initial concentrations of components during sonication. Increasing incorporation of spin label increases incorporation of cholesterol and decreases incorporation of lecithin, with the result that liposomes with cholesterol-lecithin molar ratios larger than 2 can be obtained. Besides associating with cholesterol-lecithin complexes in the liposome, the spin label seems to associate with cholesterol. Changes of the paramagnetic resonance spectrum of the liposome-bound spin label due to changes in liposomal cholesterol and spin label mole fractions - assessed by three parameters - can be used in cell-liposome interaction studies. PMID- 6276552 TI - Binding and incorporation of lecithin-cholesterol vesicles to lymphocytes: a spin label study. AB - When lecithin-cholesterol vesicles, containing the membrane-bound spin probe 3 doxyl-cholestane, were set in contact with mouse lymphocytes, the vesicles adsorbed to the cell and vesicle-membrane components were transferred to it. The spin probe was enzymatically reduced at the inside of the cell membrane. The spin label method provided a means to determine quantitatively the extent of vesicles adsorption and vesicle-cell fusion by measuring the transfer of vesicles membrane material to the cell. This method, together with the reduction of spin label by the cell, allowed also a quantitative estimate to the extent of endocytosis during cell-liposome interaction. PMID- 6276553 TI - Properties of rat erythrocyte membrane cytoskeletal structures produced by digitonin extraction: digitonin-insoluble beta-adrenergic receptor, adenylate cyclase, and cholera toxin substrate. AB - Rat erythrocyte plasma membranes have been extracted exhaustively with digitonin at low temperature, and the residual, detergent-extracted membrane cytoskeletal material is compared to that prepared with Triton X-100 with respect to protein, glycoprotein, phospholipid, and cholesterol content. Digitonin, a weaker detergent than Triton X-100, solubilizes only 26% of the phospholipids and none of the cholesterol. SDS-polyacrylamide gel electrophoresis reveals that differences between the proteins extracted by the two detergents are primarily quantitative. In terms of functional preservation, digitonin retains in the cytoskeleton 28% of the beta-adrenergic receptor binding activity (with the balance accounted for in the supernatant), greater than 90% of the adenylate cyclase and greater than 90% of the 45,000 mol wt polypeptide cholera toxin substrate. The cytoskeletal-associated beat-adrenergic receptor retains binding properties for antagonist and agonist which are identical to those of the native membrane receptor. The digitonin-extracted cytoskeleton containing the beta adrenergic receptor may provide a useful vehicle for the reconstitution of a hormone-sensitive adenylate cyclase. PMID- 6276555 TI - Fluctuation analysis of short-circuit current in a warm-blooded sodium-retaining epithelium: site current, density, and interaction with triamterene. AB - Density and conductance of the Na-site in hen coprodeum were studied by employing fluctuation analysis of short-circuit current at sodium concentrations from 26 to 130 mM. Fluctuations of current in the frequency range 2-800 Hz were induced by triamterene, a reversible blocker of conducting epithelial Na-sites. At 130 mM Na the site density was 5.8 +/- 1.0 micrometer-2 and the site conductance was 4 pS. This conductance is equal to that of the frog skin (W. Van Driessche and B. Lindemann, 1979, Nature (London) 282:519-520). Extrapolation of site density to zero sodium renders a total of 38 +/- 28 sites micrometer-2, which is compared with other estimates for the coprodeum. The site-triamterene association and dissociation constants were 9.5 +/- 0.4 rad sec-1 micrometer-1 and 255 +/- 20 rad sec-1 and they were independent of external sodium concentration. An analysis of the affinity constant for triamterene based on the DC-short-circuit current was found to be unrelated to the external sodium concentration and identical to that obtained from fluctuation analysis indicating a noncompetitive interaction between sodium and triamterene. Due to the oxygen demand of the epithelium we have developed an experimental method using short data processing times. A new analytical approach using integration of the power density spectrum proved necessary because of low signal-to-noise ratios. PMID- 6276554 TI - Erythrosin and pH gradient induced photo-voltages in bilayer membranes. AB - Erythrosin and light flashes induce voltage transients across bilayer membranes in the presence of transmembrane pH gradients. Fast voltage transients, which rise in less than 50 nsec and fall in approximately 500 nsec, are attributed to photo-deprotonation of dye sorbed in the glycerol region of phospholipid membranes. Six other halogenated xanthene dyes induce similar effects, which apparently resulted from triplet states of monoanionic dye. No photo-effects were observed with fluorescein. PMID- 6276556 TI - Organization and evolutionary progress of a dispersed repetitive family of sequences in widely separated rodent genomes. PMID- 6276557 TI - Bacteriophage P1 site-specific recombination. I. Recombination between loxP sites. PMID- 6276558 TI - Bacteriophage P1 site-specific recombination. II. Recombination between loxP and the bacterial chromosome. PMID- 6276559 TI - Base sequence studies of 300 nucleotide renatured repeated human DNA clones. PMID- 6276560 TI - Repulsive interactions between polar and apolar atoms in globular proteins. PMID- 6276561 TI - Replication of polyoma plasmid recombinants in mouse cells. PMID- 6276562 TI - Evidence for sub-families of actin genes in Dictyostelium as determined by comparisons of 3' end sequences. PMID- 6276563 TI - Structure and expression in L-cells of a cloned H4 histone gene of the mouse. PMID- 6276564 TI - Stimulation of polyoma DNA replication in isolated nucleoprotein complexes by factors from Drosophila embryos. PMID- 6276565 TI - Drosophila melanogaster ribosomal DNA containing type II insertions is variably transcribed in different strains and tissues. PMID- 6276566 TI - Physical mapping of DNA base sequence homologies between an octopine and a nopaline Ti plasmid of Agrobacterium tumefaciens. PMID- 6276567 TI - Conserved nucleotide sequences in temporally controlled bacteriophage promoters. PMID- 6276568 TI - Cloned genes for bacteriophage T4 late functions are expressed in Escherichia coli. PMID- 6276569 TI - The organization and transcription of the galactose gene cluster of Saccharomyces. PMID- 6276570 TI - Transcriptional map of bacteriophage N4. Location and polarity of N4 RNAs. PMID- 6276571 TI - Orientation of the DNA in the filamentous bacteriophage f1. PMID- 6276572 TI - Ultrastructural localization of glucose-6-phosphatase and alkaline phosphatase in the vaginal epithelium of rat. AB - The effect of ovarian hormones on the activities of glucose-6-phosphatase and alkaline phosphatase in the vaginal epithelium was studied in immature and ovariectomized rats, using ultracytochemical techniques. Comparative studies were done on normal rats at the luteal phase and on day 14 of pregnancy. Various vaginal cells show different degrees of response to progesterone and diethylstilbestrol (DES) with regard to glucose-6-phosphatase activity. Intense glucose-6-phosphatase activity was observed in the cisternae of granular endoplasmic reticulum (rER), Golgi saccules and vesicles, and nuclear envelope of both basal cells and stromal cells of progesterone treated rats, whereas in the basal cells and stromal cells of DES-treated and control animals the enzyme was totally lacking. Detectable glucose-6-phosphatase activity was also observed, however, in the rER cisternae and Golgi complex of keratohyalin-secreting squamous intermediate cells of the vaginal epithelium of DES-treated rats. Alkaline phosphatase was also found on the limiting membranes of secretory granules of mucocytes in animals at the luteal phase and during pregnancy. DES and progesterone in the doses used did not affect alkaline phosphatase activity in the rat vagina. Overall, progesterone enhances glucose-6-phosphatase activity in basal cells of the rat vagina prior to completion of mucification. Alkaline phosphatase was found in all cells involved in mucin secretion. PMID- 6276573 TI - Characterization of rat myocardial sarcolemma. PMID- 6276574 TI - Long-term epidermal growth factor-receptor internalization and processing in quiescent human fibroblasts. AB - Epidermal growth factor is internalized into cells and concomitantly induces a massive clearance of up to 90% of its total surface receptors. The hormone receptor complex is delivered to lysosomes and degraded or inactivated. Lysosomotropic alkylamines block the degradation but not the binding or internalization of ligand-receptor complexes and thus their presence results in a marked potentiation of intracellular accumulation of epidermal growth factor. We have used these alkylamines as pharmacological tools to trap internalized 125I labeled epidermal growth factor and now report that the residual population of epidermal growth factor receptors remaining on human fibroblasts after completion of the receptor clearance process is not only accessible for ligand binding but also directs the continued internalization and degradation of this growth factor over prolonged periods of time. We also show that down regulation of epidermal growth factor receptors does not result in desensitization of cells to the mitogenic response. PMID- 6276575 TI - Effect of polybrominated biphenyls on hepatic microsomal metabolism of estrogens and uterotropic action of administered estrogen in rats. AB - Perinatal exposure to polybrominated biphenyls (PBBs) increased the hepatic microsomal metabolism of estradiol, estrone, and ethynylestradiol in vitro. Pretreatment with PBBs decreased the effect of estradiol administered exogenously on uterine estrogen cytosolic receptor concentration. The effect of exogenous estradiol on uterine weight and uterine RNA content was also reduced by perinatal exposure to PBBs. Therefore, metabolism of estrogens is altered by PBBs. PMID- 6276576 TI - Influence of dose level and methionine intake on the effects of linamarin administration to rats. AB - Rats were fed a semipurified diet providing 10% casein supplemented with methionine for 2 wk, at which time some animals received the same diet without the methionine for 4 d. Animals that received linamarin were given a single oral dose containing 500 or 250 mg per kilogram of body weight. At the higher linamarin dose all animals died within 5 h after dosing. Biochemical and physiological changes observed in these rats included severe metabolic acidosis, decreased cytochrome oxidase activities, atrial fibrillation, and decreased respiratory rates. In general, the cardiac adenosinetriphosphatase enzymes were inhibited by linamarin. None of these changes were moderated by dietary methionine supplementation. At the lower linamarin dose dietary supplementation with methionine appeared to reduce incidences of clinical toxicity signs and fatalities. No methionine effect was observed in the other biochemical and physiological measurements in rats given this amount of linamarin. The results suggest that dietary supplementation with methionine provided some protection against the toxicity of the lower level of linamarin administered. PMID- 6276577 TI - Calcium phosphate crystal formation in Escherichia coli from human urine: an in vitro study. AB - Escherichia coli with no demonstrable urease activity was inoculated into filter sterilized urine obtained from a healthy volunteer subject with no history of stone disease and then incubated at 37 degrees C. Bacteria were recovered at intervals between 1 and 10 days. Urinary pH was stable as compared to control urines and spontaneous crystal precipitation was not noted in controls. Recovered organisms were analyzed by x-ray powder diffractometry. An uncharacterized mineral phase (UMP) was first evident after 6 days. Calcium phosphate in the form of brushite and hydroxyapatite was apparent at 7 and 10 days respectively. This suggests a role for bacteria in calcium phosphate crystal formation in urine apart from urease activity and may contribute to the calcium phosphate component of urinary calculi. PMID- 6276578 TI - It's not fishy: fruit of the sea may foil cardiovascular disease. PMID- 6276579 TI - Other preventive roles explored for "fish oil'. PMID- 6276580 TI - Hepatitis A and meningoencephalitis. PMID- 6276581 TI - Coexistent multiple myeloma and primary hyperparathyroidism. AB - A patient with multiple myeloma and hypercalcemia responded to cytotoxic chemotherapy. However, hypercalcemia persisted. Because of the absence of lytic bone lesions, the presence of a low serum phosphate level, and a family history of possible primary hyperparathyroidism, the patient was evaluated for this disorder. Serum parathyroid hormone and urinary cyclic adenosine monophosphate levels were elevated. Exploration of the neck disclosed two enlarged parathyroid glands (1,850 mg and 210 mg), which were excised. After surgery, the patient's serum calcium levels remained normal for one year. Progressive myeloma bone disease developed that eventually resulted in recurrent hypercalcemia and death. Autopsy revealed only evidence of myeloma. PMID- 6276582 TI - Adrenocorticotropic hormone, beta-lipotropin, and endorphin-related peptides in health and disease. PMID- 6276583 TI - Two cases of low-renin hypertension thought to be due to excessive secretion of unknown mineralocorticoid. AB - Two patients with low renin hypertension showing an increased urinary excretion with 17-KS, with normal level of plasma deoxycorticosterone and no signs of virilization were reported. Dexamethasone induced reduction in blood pressure and elevation of serum K, in spite of acceleration of the renin-angiotensin aldosterone system. Thus, it has been inferred that the hypertension was not associated with adrenogenital syndrome but was due to excessive production of an unknown mineralocorticoid. PMID- 6276584 TI - [Effects of intrathecal beta-endorphin and morphine on post-operative pain (author's transl)]. PMID- 6276585 TI - [Effects of epidural injection of beta-endorphin on endocrine function in man (author's transl)]. PMID- 6276587 TI - [Incidence and survival of Clostridium perfringens in soil (author's transl)]. PMID- 6276588 TI - [Platelets (author's transl)]. PMID- 6276586 TI - [Clinical evaluation of pivmecillinam in the maintenance therapy of chronic urinary tract infection (author's transl)]. AB - In order to evaluate the efficacy and safety of pivmecillinam (melysin tablet, PMPC), PMPC was administered to 78 chronic UTI cases in the field of obstetrics and gynecology (posthysterectomy infection, chronic cystitis, chronic pyelonephritis and etc.). In principle, daily 400 mg of PMPC was administered for 2 weeks. (1) Overall clinical efficacy judged by doctor was evaluated in 78 cases and the result was; excellent in 17, good in 37, fair in 10, poor in 13 and unknown in 1 case with the effectiveness rate of 69.2%. (2) Overall clinical efficacy judged by 'criteria for clinical evaluation in complicated UTI' recommended by UTI study member was evaluated in 54 cases and the result was; excellent in 15, good in 20 and poor in 19 cases with the overall efficacy rate of 64.8%, the result of which was similar to that of doctor's judgement. (3) Efficacy on pyuria was evaluated in 72 cases and it was cleared in 27, decreased in 25, unchanged in 20 and unknown in 6 cases. Efficacy on bacteriuria was evaluated in 72 cases and it was eliminated in 44, decreased in 9, replaced in 8, unchanged in 8 and unknown in 9 cases. (4) Side effect, considered by doctors to be caused by PMPC administration, was noticed in 3 out of 78 cases (3.8%), all of which was mild gastrointestinal disturbance and the administration of PMPC was continued. Abnormal change of laboratory finding considered by doctors to be caused by PMPC administration was noticed in 1 out of 78 cases, which was slight elevation of GOT and GPT values. It is therefore considered that PMPC appear to be useful drug for the maintenance therapy of chronic UTI in the field of obstetrics and gynecology. PMID- 6276589 TI - [An autopsy case of smoldering acute leukemia with double gastric cancers (author's transl)]. PMID- 6276590 TI - [Hepatic collagenase activity as an indicator of hepatic fibrosis (author's transl)]. PMID- 6276591 TI - [Electronmicroscopic observations of necrolytic migratory erythema in glucagonoma (author's transl)]. PMID- 6276592 TI - [Histochemical and biochemical studies on stress ulcer in water immersion restrained rat (author's transl)]. PMID- 6276593 TI - [Experimental induction of Mallory bodies in mice by griseofulvin treatment (author's transl)]. PMID- 6276594 TI - [A case of a metastatic hepatocellular carcinoma to the stomach via blood stream (author's transl)]. PMID- 6276595 TI - [Studies on autofluorescense of hepatoma excited by argon laser light (author's transl)]. PMID- 6276596 TI - [Clinical significance of CEA in ascites (author's transl)]. PMID- 6276597 TI - [Changes of plasma amino acids levels in liver diseases (author's transl)]. PMID- 6276598 TI - [An autopsy case of hepatocellular carcinoma following transcatheter arterial embolization (author's transl)]. PMID- 6276599 TI - [Mailmen's vibration hazards induced by motorcycle riding (author's transl)]. PMID- 6276600 TI - A survey of rotavirus infection in the topics. AB - The age distribution of rotavirus antibody in the serum of inhabitants of Laos and Indonesia was studied by the neutralization test using antigenically related calf rotavirus (NCDV) as a substitute for non-cultivable human rotavirus. The results revealed that both the rate of antibody-positives and the modal titers of antibody distribution by each age group in these countries were higher than those in respective are groups of Japanese, which suggest higher endemicity of rotavirus infection in these countries. A survey of rotavirus in diarrheic patients' stools by enzyme-linked immunosorbent assay indicated that the virus infection occurred most frequently among infants between 4 and 12 months of age. PMID- 6276601 TI - Chemosensitive neuron in the hypothalamus related to food intake behavior. PMID- 6276602 TI - Electropharmacological actions of catecholamine in sympathetic ganglia. Multiple modes of actions to modulate the nicotinic transmission. PMID- 6276603 TI - Hepatoma in children--a clinical analysis on 15 cases. AB - In ten boys and five girls with hepatoma, evaluations were made as to the sex, age, pathologic findings, operation and prognoses. Ages at the time of operation ranged from 1 month to 12 years. In twelve, the tumors were successfully removed; 4 underwent left lateral segmentectomy, 4 right hepatic lobectomy and 4 trisegmentectomy. At least at the time of completion of this paper, eight of these 12 patients (66%) were alive including one patient with pulmonary metastases. Thirteen patients had hepatomas of massive type and 2 had nodular type. Histologic type of these hepatomas was hepatoblastoma in 14 cases and hepatocarcinoma (liver cell carcinoma) in one. These hepatoblastomas included 10 of the fetal type and 4 of the embryonal type. In five of 12 patients pulmonary metastases were detected after the hepatectomy. Four of these 5 patients underwent pulmonary surgery; one girl is alive with no sign of recurrence for over 8 years after surgery on both lungs. Histological examination of pulmonary lesions disclosed hepatoblastomas of an embryonal type, and such were not related to the histological types of hepatic lesions. PMID- 6276604 TI - Coated polyglactin 910--a new synthetic absorbable suture. PMID- 6276605 TI - [Effects of cigarette smoking on pulmonary functions and on plasma levels of serotonin, histamine and serum angiotensin I converting enzyme activity (author's transl)]. PMID- 6276606 TI - [Angiotensin I converting enzyme activity in patients with spontaneous pneumothorax and in rabbits with artificial pneumothorax (author's transl)]. PMID- 6276607 TI - [Two cases of retroperitoneal non-chromaffin paraganglioma with symptoms of anuria (author's transl)]. PMID- 6276608 TI - Serodiagnosis for feline infectious peritonitis by immunofluorescence using infected suckling mouse brain sections. PMID- 6276609 TI - The presence of B type virions in trichoepithelioma of a mouse. PMID- 6276610 TI - Prevalence of antibodies against Getah virus in horses raised in Hokkaido. PMID- 6276611 TI - [Tuberactinomycin-N (enviomycin-EVM) in the treatment of pulmonary tuberculosis (author's transl)]. PMID- 6276612 TI - Adenosine triphosphate facilitates the Na+-K pump of frog skeletal muscle fibres. PMID- 6276613 TI - A study of the photobiogenesis of cholecalciferol in vivo and the constraint on its 25-hydroxylation in rat. PMID- 6276614 TI - Localization of 4-ene-5alpha-reductase, 17 beta-ol-dehydrogenase and aromatase in immature rat ovary. PMID- 6276615 TI - Renal pseudotumour following treatment of Wilms' tumour. AB - A 9 1/2-year-old boy was found to have a mass occupying the lower pole of his left kidney, six years after his right kidney had been removed because of a Wilms' tumour. Investigation of the mass included angiography which demonstrated a mass with pathological arteries. Open biopsy of the mass revealed histological changes of nonspecific nephritis. It is probable that this nephritis was a result of radiation damage potentiated by concurrent treatment with actinomycin D, which he had received six years ago. PMID- 6276616 TI - Epidemiologic and prognostic factors for lung cancer in a county hospital. AB - This is a study of 269 patients with lung cancer at the Erie County Medical Center who were admitted between 1973 and 1978. They were analyzed for sex, race, age, history of smoking, occupation, tumor cell type, cytology, incidence of metastases, changes in liver function, mode of treatment, and survival. The incidence of cancer was highest in white males. Only 1.5% of patients were under age 40. Smoking was a predisposing factor. Not enough information was available to determine the relationship of occupation to lung cancer. Squamous cell carcinoma was the most common (53.9%), followed by adenocarcinoma (16.0%) and small-cell carcinoma (12.6%). Sputum cytology was 28.3% sensitive, and bronchial washings were 52.2% sensitive. A greater incidence of bone metastases from a small-cell primary (50%) was found than is reported in the literature. Changes in SGOT and/or SGPT liver enzymes correlated significantly with liver metastases, but not with tumor cell type. Changes in alkaline phosphatase correlated well with bone metastases. Radiation was the most commonly used mode of therapy. The best survival was achieved in patients treated by surgery (22.6 months), followed by surgery and radiation (16.2 months); those treated by radiation alone had a mean survival time of 8.7 months. Untreated patients had a mean survival time of 2.4 months. Treated patients with adenocarcinomas had the longest survival (18.5 months), compared to 13.0 months for those with squamous cell carcinomas and 8.4 months for those with small-cell carcinomas. Only three patients survived 5 years, all of whom were treated surgically for adenocarcinoma. No patients with tumors of other cell types survived 5 years. The 5-year survival rates are 2.1% (3 of 141) for all histologic types of lung cancer and 12.5% (3 of 24) for adenocarcinoma. PMID- 6276617 TI - Gastric duplication associated with adenomyoma. AB - A case of gastric duplication associated with adenomyoma including ectopic pancreatic tissue in an adult patient is reported. The histologic features, diagnostic criteria, and pathogenesis are briefly discussed. PMID- 6276618 TI - Intramembrane events during the nerve impulse. PMID- 6276619 TI - On the chemiosmotic hypothesis and the nature of the mitochondrial protonmotive force. PMID- 6276620 TI - Prosthetic replacement of entire left hemidiaphragm in malignant fibrous histiocytoma of the diaphragm. AB - A case of primary malignant fibrous histiocytoma of the diaphragm which, to our knowledge, is the first record in the world literature, is presented. It occupied almost the entire left hemidiaphragm and was surgically removed with the left lower pulmonary lobe, stomach, colon, and spleen. The entire left hemidiaphragm then was replaced with Marlex mesh. Prosthetic replacement of the entire hemidiaphragm, which has not been previously reported, was successfully performed with preservation of pulmonary function on the affected side. The patient died of brain metastases of malignant fibrous histiocytoma more than 4 months postoperatively. Diagnosis of diaphragmatic tumors and a technique which involves the use of a prosthesis are discussed. PMID- 6276621 TI - [Lymphocytes aminergic binding changes in chronic lymphocyte leukaemia]. PMID- 6276622 TI - High levels of 5'-nucleotidase activity in blastic chronic myelogenous leukemia with common ALL-antigen. PMID- 6276623 TI - Microforge modifications useful for construction of micropipettes to record the response of single ionic channels from living cells. PMID- 6276624 TI - Quantitative densitometry of neurotransmitter receptors. AB - An autoradiographic procedure is described that allows the quantitative measurement of neurotransmitter receptors by optical density readings. This procedure is a modification of the method of Young and Kuhar (1979a). Frozen brain sections are labeled in vitro with [3H]ligands under conditions that maximize specific binding to neurotransmitter receptors. The labeled sections are then placed against the 3H-sensitive LKB Ultrofilm to produce the autoradiograms. These autoradiograms resemble those produced by [14C]deoxyglucose autoradiography (Sokoloff, 1977) and are suitable for quantitative analysis with a densitometer. Muscarinic cholinergic receptors in rat and zebra finch brain and 5-HT receptors in rat brain were visualized by this method. When the proper combination of ligand concentration and exposure time are used, the method provides quantitative information about the amount and affinity of neurotransmitter receptors in brain sections. This was established by comparisons of densitometric readings with parallel measurements made by scintillation counting of sections. PMID- 6276625 TI - Effects of cyproheptadine and metyrapone on ACTH and aldosterone concentrations in patients with Cushing's disease: a preliminary report. PMID- 6276626 TI - [Non-secreting Langerhans carcinoma in children]. PMID- 6276627 TI - [Peripheral neuropathy in hemodialysis (author's transl)]. PMID- 6276628 TI - [Chest radiography in hepatocellular carcinoma (author's transl)]. PMID- 6276629 TI - Pharyngoesophageal swallowing disorders and the pharyngoesophageal sphincter. PMID- 6276630 TI - [Hypothalmo-pituitary-adrenal function--can a short ACTH-test replace insulin tolerance test?]. PMID- 6276631 TI - [Captopril inhibits angiotensin converting enzyme. New principles for therapy in hypertension and heart failure]. PMID- 6276632 TI - [Hereditary pyrophosphate arthropathy in 3 Swedish families]. PMID- 6276633 TI - Buprenorphine : characteristics of binding sites in the rat central nervous system. PMID- 6276634 TI - Cyclic AMP and cyclic GMP as the respective mediators of the intracycle stimulation of DNA synthesis and mitosis induced by glucagon and insulin in primary neonatal rat hepatocytes. PMID- 6276635 TI - Purification of calmodulin from human brain. PMID- 6276636 TI - Dietary sodium regulation of blood pressure and renal alpha 1- and alpha 2 receptors in WKY and SH rats. PMID- 6276638 TI - Hepatitis A infection: guidelines for development of satisfactory assays for laboratory diagnosis. PMID- 6276637 TI - Effect of buffer constituents on rat liver 3-hydroxy-3-methyl glutaryl coenzyme A reductase. AB - Rat liver microsomes prepared in Tris buffer exhibited 3 to 10 times higher 3 hydroxy-3-methyl glutaryl CoA reductase specific activity than microsomes prepared with potassium phosphate buffer. This higher activity was observed in rats killed during mid-light cycle, but microsomes from rats killed during mid dark cycle showed no significant difference in enzyme activity between buffers. When microsomes prepared in the 2 different buffers were preincubated with ATP and MG++, enzyme activity was inhibited to the same extent. The cytosol fraction in each of the 2 different buffer preparations possessed similar phosphatase activity. The higher 3-hydroxy-3-methyl reductase activity in Tris buffer, therefore, does not appear to be due to differences in phosphorylation or dephosphorylation activity. PMID- 6276639 TI - Diagnosis of acute hepatitis a infection: protein A absorption-an extended trial. PMID- 6276640 TI - Human monoclonal antibodies: their technology and application. PMID- 6276641 TI - [Osteoscintigraphy in the diagnosis of skull bone defects of various origins]. PMID- 6276642 TI - [Interstitial beta therapy of cystic craniopharyngiomas]. PMID- 6276643 TI - [Bone scintigraphy with 99mTc-pyrophosphate in the diagnosis of the degree of the spread of the tumor process in the head and neck area]. PMID- 6276644 TI - [The clinical value of hormone neutralization by antibodies]. PMID- 6276645 TI - [A new preparation for peripheral arterial occlusive diseases]. PMID- 6276646 TI - Isolated ACTH deficiency: a heterogeneous disorder. Critical review and report of four new cases. AB - Isolated adrenocorticotropin (ACTH) deficiency is a rare cause of secondary adrenocortical insufficiency. This review summarizes the clinical and laboratory features of 39 previously reported cases plus 4 new patients. The clinical manifestations of isolated ACTH deficiency are variable, nonspecific and similar to those seen in adrenocortical insufficiency of any cause. The diagnosis of isolated ACTH deficiency due to intrinsic pituitary disease is made unequivocally when all the following criteria are met: 1) low basal urinary 17 hydroxycorticosteroid (17-OHCS) levels with or without low basal plasma cortisol, 2) low or normal basal plasma ACTH, 3) stimulation of cortisol, 17-OHCS or both during prolonged ACTH administration, 4) lack of 17-OHCS elevation in response to metyrapone and 5) normal secretory indices of other pituitary hormones. Isolated ACTH deficiency secondary to suprapituitary (e.g., hypothalamic) dysfunction is also based upon the above criteria, but, in addition, is associated with stimulation of cortisol and ACTH secretion following vasopressin administration. PMID- 6276647 TI - Hypoglycemic peripheral neuropathy in association with insulinoma: implication of glucopenia rather than hyperinsulinism. Case report and literature review. AB - A syndrome of peripheral polyneuropathy associated with islet cell tumors and hypoglycemia has been reported in 28 patients. Despite varying features in these patients, the clinical characteristics of this syndrome are remarkably similar. These consist of the development of a sensorimotor neuropathy during a protracted course of recurrent severe hypoglycemia, related to underlying insulinoma. Cerebral symptoms dominate the clinical picture and a predominantly or entirely motor, distal and symmetric, peripheral neuropathy ensures. Upper limb involvement is more frequent, accompanied by severe weakness and distal wasting, usually without fasciculations. Painful distal paresthesias without objective sensory loss are characteristic. Direct relationship to a single hypoglycemic insult is often absent. This report describes the clinical features and laboratory investigation of a new case with this condition, reviews the literature and discusses the syndrome with special regard to the etiology. PMID- 6276649 TI - The study of hemoglobin by electron paramagnetic resonance spectroscopy. PMID- 6276648 TI - Carcinomatous leptomeningitis in small cell lung cancer: a clinicopathologic review of the National Cancer Institute experience. AB - CLM developed in 60 of 526 patients (11%) with SCLC seen at the NCI between August 1969 and June 1980. Life table analysis revealed an overall 25% risk of CLM at 3 years. CLM was diagnosed during all phases of the patients' clinical course, but the majority (83%) were cases diagnosed at the time of progressive systemic disease. Univariate log rank analysis indicated that pretreatment factors associated with the development of CLM included: involvement of the brain, spinal cord, bone marrow, liver or bone; extensive disease; and male sex. Patients who did not obtain a complete response to systemic therapy were at greater risk of developing CLM than complete responders. Multivariate analysis of these factors indicated that liver metastases were most strongly associated with the time to development of CLM, followed in order of importance by bone and CNS metastases. Patients usually presented with signs and symptoms reflecting involvement of multiple areas of the neuraxis including the cerebrum, cranial nerves and spinal cord; 51 of the 60 patients had intracerebral metastases and 27 had spinal cord lesions during their clinical course. Autopsy features including focal or diffuse involvement of the leptomeninges with infiltration of the Virchow-Robin spaces were similar to meningeal lymphoma and leukemia, except that CLM was rarely the sole manifestation of CNS tumor. Median survival following the diagnosis of CLM was 7 weeks. However, most deaths were attributed to systemic disease, and treatment with intrathecal chemotherapy and irradiation often provided palliation. With the increased awareness of this complication, an antemortem diagnosis increased from 39% prior to 1977, to 88% of patients after 1977. PMID- 6276650 TI - Magnetic susceptibility of hemoglobins. PMID- 6276651 TI - DNA analysis in the diagnosis of hemoglobin disorders. PMID- 6276652 TI - Preparation of mammalian hepatocytes. PMID- 6276653 TI - Thiol S-methyltransferase. PMID- 6276654 TI - Arylamine N-methyltransferase. PMID- 6276655 TI - Benzoyl-CoA: amino acid and phenylacetyl-CoA: amino acid N-acyltransferases. PMID- 6276656 TI - S-Formylglutathione hydrolase. PMID- 6276657 TI - Preparation and characterization of bluetongue virus. PMID- 6276658 TI - Preparation and characterization of encephalomyocarditis (EMC) virus. PMID- 6276659 TI - Assay of (2'-5')-oligoadenylic acid synthetase levels in cells and tissues: a convenient poly(I) . poly(C) paper-bound enzyme assay. PMID- 6276660 TI - Radioimmune and radiobinding assays for A2'p5'A2'p5'A, pppA2'p5'A, and related oligonucleotides. PMID- 6276661 TI - Assay of (2'-5')-oligo(A) synthetase with 2',5'-ADP-Sepharose. PMID- 6276662 TI - Glycosylation steps involved in processing of pro-corticotropin-endorphin in mouse pituitary tumor cells. PMID- 6276663 TI - Role of the 6-phosphomannosyl-enzyme receptor in intracellular transport and adsorptive pinocytosis of lysosomal enzymes. PMID- 6276664 TI - The role of microtubules and microfilaments in lysosomal enzyme release from polymorphonuclear leukocytes. PMID- 6276665 TI - Import of proteins into mitochondria. PMID- 6276666 TI - The "secretory code" of the neutrophil. PMID- 6276667 TI - Aspects of the calcium hypothesis of stimulus-secretion coupling: electrical activity in adenohypophyseal cells, and membrane retrieval after exocytosis. PMID- 6276668 TI - Synthesis and assembly of transmembrane viral and cellular glycoproteins. PMID- 6276669 TI - Relationship between receptors, calcium channels, and responses in exocrine gland cells. PMID- 6276670 TI - Chromosomal loci of genes controlling site-specific restriction endonucleases of Bacillus subtilis. AB - We constructed transformation of B. subtilis 168 which acquired genes for site specific restriction endonucleases. These endonucleases originated from various strains of B. subtilis and were classified into five groups based on the specificity of the sequences recognized by the enzymes. We examined the loci of genes for site-specific restriction endonucleases belonging to different groups: hsrE determined Endo. R. Bsu1231 (I), hsrB Endo.R.Bsu1247(I), hsrR Endo.R.BsuR and hsrC Endo.R.Bsu-1247(II). One gene, hsrE, was located between sacA and purA by transduction crosses with phage PBS1, and another gene, hsrB, between hsrE and purA. Genes hsrR and hsrC had been suggested to be allelic or closely linked by previous studies with transformation. We located hsrR and hsrC between purB and tre. Our previous observation and this study show that B. subtilis 168 has at least three independent loci on the chromosome for four genes for site-specific restriction endonucleases in addition to the locus for the original restriction activity (Bsu168-specific restriction) of strain 168. PMID- 6276671 TI - Inactivation of the Serratia marcescens gene for the lipoprotein in Escherichia coli by insertion sequences, IS1 and IS5; sequence analysis of junction points. AB - A pBR322-derived plasmid pKEN221 carrying a Serratia marcescens lpp gene overproduces the outer membrane lipoprotein in an Escherichia coli lpp- cell. However, when this strain was continuously cultured in a rich medium for about thirty generations, many Lpp- mutants were accumulated. Out of six mutants analyzed, three were found to carry insertion mutation in the lpp gene in pKEN221. From resistance enzyme mapping and hybridization analysis of the mutant plasmid DNA, it was found that two mutants were caused by insertion sequence IS1 and one by IS5. Nucleotide sequence analysis of these mutant DNAs revealed that both IS1 and IS5 insertions occurred in the A-T rich 5' untranslated region of the lpp gene. While the IS1 insertion resulted in a direct duplication of a nine base-pair sequence in the original pKEN221 DNA at the junction with IS1, the IS5 insertion resulted in a direct duplication of a four-base-pair sequence. IS5 was found to contain inverted-repeat sequences of twelve nucleotides at its exact ends. This is the first example of the nucleotide sequence analysis of an IS5 insertion mutation. By Southern blot hybridization, the E. coli chromosomal DNA was found to contain about ten copies of IS5. PMID- 6276672 TI - The I--R system of hybrid dysgenesis in Drosophila melanogaster: are I factor insertions responsible for the mutator effect of the I--R interaction? AB - When Drosophila melanogaster males coming from a class of strains known as inducer are crossed with females from the complementary class (reactive), a quite specific kind of sterile female (SF) is obtained that exhibits other dysgenic traits such as non-disjunctions and non-randomly distributed mutations. This syndrome is caused by the interaction of the 'I factor' linked to inducer chromosomes with the maternally inherited reactive state 'R'. This I--R interaction is also responsible for 'chromosomal contamination' that is likely to result from very frequent I factor insertions into reactive chromosomes. Such insertions might be responsible for the I--R induced mutations and some data concerning this hypothesis are reported here. Out of nine I--R-generated mutants one, the whiteIR1 (wIRI) allele, is closely linked to an I factor, which maps either at the site of the mutation or within less than 0.02 map units. In addition, wIR1 is somewhat unstable when transmitted through SF females. In contrast, the typical I factor does not seem to be associated with any of the eight other mutants as judged by their inability to induce the female sterility characteristic of the I--R syndrome. The possibility is discussed that most of I- R-induced mutations are nevertheless caused by insertions of either undetectable I factors of other transposable elements, not related to I, whose transposition is dependent on the I--R interaction. PMID- 6276673 TI - Essential role of the gyrB gene product in the transcriptional event coupled to dnaA-dependent initiation of Escherichia coli chromosome replication. AB - When a culture of the gyrB41-ts mutant is shifted to the nonpermissive temperature, DNA synthesis is arrested at the initiation phase of chromosome replication. After thermal inactivation of the gyrB gene product reinitiation occurs in the presence of chloramphenicol but not in the presence of rifampicin. This suggests that the B subunit of DNA gyrase may regulate synthesis of an "initiator RNA". An rpoB202 mutation has been isolated which suppresses both the DnaA-initiation phenotype and the inhibitory action of antibiotics which are known to result in relaxation of chromosomal DNA in vivo. We propose that DNA tertiary structure rather than DNA gyrase itself plays an essential regulatory function in the dnaA-dependent transcription which precedes the initiation of chromosome replication. PMID- 6276674 TI - Apurinic DNA endonucleases from Drosophila melanogaster embryos. AB - Apurinic DNA endonuclease activity from Drosophila melanogaster embryos was resolved into two separable forms by phosphocellulose chromatography, one which flowed through the column (Fraction I) and the other which was retained and eluted at approximately 200 mM potassium phosphate (Fraction II). Both fractions, purified further by glycerol gradient sedimentation, were found to introduce nicks into DNA that were specific for and equal in number to the alkali-labile sites in depurinated DNA. They had similar apparent Km values for apurinic sites (0.7 nM apurinic sites for Fraction I and 0.8 nM for Fraction II), but differed with respect to optimal pH, Mg++ requirement and sensitivity to EDTA. PMID- 6276675 TI - Recombinational instability of F' plasmids in Escherichia coli K-12: localization of fre-sites. AB - The F' plasmids ORF-1 (purE+ tsxs proC+ lac+) and F'14 (argE+ metB+ ilv+) contain active regions of recombination, fre I and fre II correspondingly. The plasmid ORF-1 is stable in recF- cells (i.e., with the RecBC pathway of recombination) and decays in rec+ cells (RecBCF pathway) giving two types of product: F+ and plasmid pCK-1 (tsxs proC+ lac+) containing part of the initial DNA. They are extremely instable in the presence of the RecF pathway, (recBC- sbcB-), yielding F+ and plasmid pCK-2 (proC+ lac+). The instability of plasmids depends on a region of homology between the chromosome and the episome. The instability of ORF 1 shows the participation of IS3 elements (alpha 1 beta 3 and alpha 3 beta 1) in the recA, recF-dependent recombinational decay and allows localization of two active sites on the chromosome: fre I1 between purE and tsx markers and fre I2 between tsx and proC. The plasmid F'14, in accordance with published data, is able to yield F+ cells by recA-independent recombination. But eventually this plasmid may undergo a recA, recF-dependent decay. Genetic analysis of these events allows localization of an active point of recombination, freII1, between argE and metB. Another active point is localized inside the F factor. The recA dependent decay of plasmid F-14 is also excluded on the RecBC pathway (recF- strains). PMID- 6276676 TI - Relation between the transforming activity of a marker and its proximity to the end of the DNA particle. AB - Transforming pneumococcal DNA is inactivated by treatment with restriction enzymes. For mutations belonging to the same locus (amiA locus), the extent of inactivation depends strongly upon the mutations and the enzymes. Two EcoRI and one BamHI restriction sites have been located within the amiA locus. After treatment of donor DNA with either one of these enzymes, the lowest transforming activity is observed for mutations that map near restriction sites. This effect of proximity to the nearest end of the DNA fragment extends over a distance of 1,400 nucleotides. The curve of transforming activity versus DNA size obtained with endonuclease-generated DNA fragments is very similar to that obtained previously with mechanically sheared DNA. Both curves show a striking slope change for donor DNA size around 2,700 base pairs, i.e. twice the length found for the extent of the 'end effect'. We suggest that for donor DNA fragments larger than 2,700 base pairs the transforming activity depends mainly upon the size of donor whereas for donor DNA fragments shorter than 2,700 base pairs both a size-dependent phenomenon and the 'end effect' contribute to reduce drastically the transforming activity. PMID- 6276677 TI - Integration specificity of an artificial kanamycin transposon constructed by the in vitro insertion of an internal Tn5 fragment into IS2. AB - IS2 has been marked genetically by the in vitro insertion into its HindIII site of a 3.3 Kb HindIII fragment of Tn5 conferring resistance to kanamycin. The transposition of the IS2::Km, thus obtained, to lambda has been found and insertion sites were characterised. Each of ten independent IS2::Km insertions were found at the same site at 61.2% of the lambda map, always in the same orientation (orientation II relative to the xis gene). The integration sites of IS2::Km in five of the kanamycin-transducing phages were determined by DNA sequence analysis, and were found to be identical at the nucleotide level. Further transposition of IS2::Km from lambda to the bacterial chromosome was demonstrated. PMID- 6276678 TI - Structure and function of the region of the replication origin of the Bacillus subtilis chromosome. I. Isolation and characterization of plasmids containing the origin region. AB - A BamHI restriction endonuclease fragment, B7, which is replicated first among all other fragments derived from the Bacillus subtilis chromosome, was cloned in Escherichia coli using as vector a hybrid plasmid pMS102 that can replicate both in E. coli and B. subtilis. Digestion of pMS102 with BamHI produced two fragments and the smaller one was replaced by the B7 fragment. The cloned plasmid pMS102' B7 exhibited some peculiar properties that were not observed with plasmids containing other fragments from the B. subtilis chromosome. (1) E. coli cells harboring this plasmid stuck to each other and to glass. This property was more apparent when cells were grown in poor media. (2) E. coli cells tended to lose the plasmid spontaneously when they were grown without the selective pressure favorable to the plasmid. (3) The frequency of transformation of B. subtilis by pMS102'-B7 was less than 1/1,000 of that by the vector plasmid pMS102'. The number of copies of pMS102'-B7 present in the transformants was also markedly reduced, although the pUB110 origin of replication on the vector was intact and should be functional in B. subtilis. This inhibitory effect of the B7 fragment on plasmid replication was confirmed more directly by developing a semi in vitro replication system using protoplasts. Both in E. coli and B. subtilis the B7 fragment affected replication of its own molecule but not that of the coexisting plasmid with an identical replication system. The implication of the function of the B7 fragment in the initiation of the B. subtilis chromosome will be discussed. PMID- 6276680 TI - Genetic and nucleotide sequence homologies in bacillus genomes. AB - The extent of divergence in the organization of the aromatic amino acid cluster among the heterogenetic strains of Bacillus subtilis has been examined by hybridizations to a trp homolog from B. pumilus and by marker survivals after restriction. The trp operon in the W23, 3610 and 168M genomes exhibit variations in the location of the ECoRI and HindIII cleavage sites consistent with the relative transforming activity of the surviving genes and the history of the strains. PMID- 6276679 TI - Structure and function of the region of the replication origin of the Bacillus subtilis chromosome. II. Identification of the essential regions for inhibitory functions shown by the DNA segment containing the replication origin. AB - A BamHI restriction endonuclease fragment B7, which contains the replication origin of the Bacillus subtilis chromosome, showed inhibitory effects on cell growth and plasmid replication in Escherichia coli and Bacillus subtilis, when B7 was inserted into a composite plasmid pMS102' and introduced into these cells. In order to localize these properties in more limited regions within the B7 fragment, we developed a new and widely applicable method for deletion of DNA segments of various lengths from one or other end of a given region of the plasmid molecule. Using a set of deletions in the B7 fragments of pMS102'-B7, we determined the loci responsible for the inhibitory effects of B7 as described below. (1) Stickiness appearing in E. coli cells was caused by a segment residing in a region of approximately 2.2 kilobase pairs (kbp) overlapping the E19 and E22 fragments. (2) instability of the plasmid in E. coli was due to a segment localized in the 440 bp region of the E19-side terminal portion of the 2.2 kbp region. (3) The same 440 bp were also responsible for inhibition of the replication of the plasmid in B. subtilis. Hybridization of the cloned DNA fragments containing the 2.2 kbp region with the whole B. subtilis chromosome revealed that several regions of the chromosome are homologous to this characteristic sequence. PMID- 6276681 TI - Further insight on the transferred-DNA of octopine crown gall. AB - Six octopine tumour lines incited by pTiB6S3, pTiAch5 and pTiA6 on tobacco, Arabidopsis and Petunia were studied by the Southern blotting hybridisation technique in order to define accurately the dimensions of the segments of plasmid origin transferred to the tumourous cell and their organisation in the plant genome. Emphasis has been put on the comparison between octopine and nopaline T DNAs and on the lines presented here compared with those studied previously (Thomashow et al. 1980). The length of the transferred DNA segment does not depend on the plasmids used, nor on the host plants. The octopine T-DNA organisation in the cell nucleus is significantly different from that of nopaline T-DNAs: tandem arrangements of T-DNA segments could not be detected and the T-DNA itself is much shorter. The tumour lines described here can be compared to some extent with those studied by another group (Thomashow et al. 1980) by the same technique. However, some differences were observed. The transferred DNA was seen as a unique stretch of about 11 kb present only once per cell. No amplification of any part was noticed in any of these six lines. Examination of the restriction patterns presented by the boundary fragments of the T-DNA in these lines suggested that some of them were of common origin. PMID- 6276682 TI - Cloning of E. coli pnp gene from an episome. AB - Starting with an F' episome harboring a transposon inserted in the pnp gene (Portier 1980), we were able to identify an EcoRI restriction fragment carrying the pnp and argG genes. This fragment, from both wild-type and mutant episomes, was cloned ni pACYC184. The presence of argG on the fragment allowed positive selection of the desired clones in an auxotrophic strain (argG). A restriction map was established and a fragment of 3 megadaltons subcloned in the plasmid vector pBR322. The pnp gene corresponds to about 50% of this subcloned segment and was roughly located by deletion mapping. The direction of transcription and locations of the promotor and gene extremities were determined by analyzing proteins synthesized in "maxi-cells". In addition, the gene coding for a 10,000 dalton protein was found to reside adjacent to the beginning of the pnp structural gene. Strains carrying plasmids which express the pnp overproduce polynucleotide phosphorylase. PMID- 6276683 TI - Protein H encoded by plasmid Clo DF13 involved in lysis of the bacterial host. I. Localisation of the gene and identification and subcellular localisation of the gene H product. AB - The gene expression of the Clo DF13 "replication region", located between 1.8% and 12% on the plasmid genome, was studied using newly constructed Clo DF13 insertion and deletion mutants. We were able to detect a Clo DF13 specified protein of 6 kilodaltons (kd) by electrophoretic analysis of plasmid proteins, synthesized in Escherichia coli minicells, on 14-25% gradient polyacrylamide gels. The gene encoding this protein was mapped between 1.8% and 12% on the Clo DF13 genome. The nucleotide sequence of this region, as determined by Stuitje et al. (1980), revealed three open reading frames each potentially coding for a protein of 6 kd. Since these three proteins differ in amino acid composition we could distinguish which of these proteins was actually synthesized, by labeling Clo DF13 proteins with specific 14C-labeled amino acids. We found that gene H, located between 9.3% (bp 744) and 11% (bp 893), encodes the observed protein of 6 kd (denominated protein H). With respect to the subcellular localization we observed that protein H, which contains a large hydrophobic region at its C terminal part, is predominantly present in the bacterial membrane. Although gene H is located close to the region known to be involved in Clo DF13 replication, its gene product, protein H, is not essential for the plasmid DNA replication process. The possibility of the existence of a comparable protein encoded by the related plasmid Col E1 will be discussed. PMID- 6276684 TI - Isolation and properties of Tn10 insertions in the rac locus of Escherichia coli. AB - Two Tn10 insertions that are in the rac locus of the chromosome of Escherichia coli have been isolated and characterized. The insertions are located at min 29.7 and min 30.0. The insertions are stable when an F123 rac::Tn10 episome is transferred to an F- rac+ recipient, but they are lost at a high frequency when transferred to an F- rac- recipient. This latter condition has been previously demonstrated to cause the excision of the rac locus. The Tn10 insertions are also lost at a high frequency when strains containing them are lysogenized with lambda reverse. If the lysogens that have lost the Tn10 insertion are subsequently cured of lambda reverse, the cells no longer contain sequences homologous with rac locus DNA. These strains were rac- when tested for recombination activation (Low 1973), and this procedure consequently provides a simple means to make isogenic rac+ and rac- strains. PMID- 6276685 TI - DNA repair in E. coli strains deficient in single-strand DNA binding protein. AB - Weigle reactivation and mutagenesis have been found to be defective in strains of E. coli deficient in single-strand DNA binding protein (SSB). These defects parallel those previously found in prophage induction and amplification of recA protein synthesis in ssb- strains. Together, these results demonstrate a role for SSB in the induction of SOS responses. UV survival studies of ssb- recA- and ssb- uvr- strains are presented which also suggest a role for SSB in recombinational repair processes but not in excision repair. Studies of host cell reactivation support this latter conclusion. PMID- 6276687 TI - The effect of tnm mutations of Escherichia coli K12 on transposition of various movable genetic elements. AB - The effects of different tnm mutations on the transposition of Tn3, Tn5, Tn10, Tn601, and bacteriophage Mu were studied. Five tnm mutations were placed into three phenotypic classes. The representatives of the first class, tnm-1 and tnm 2, caused a complete block of transposition of all Tn-elements studied; the representatives of the second class, tnm-3 and tnm-4, specifically affected Tn9 transposition while the tnm-5 mutation attributed to the third class caused formation of cointegrates between the donor genome of phage lambda carrying Tn elements Tn3 or Tn5 and the recipient genome (bacterial chromosome). PMID- 6276686 TI - Variable expression of the ssb--1 allele in different strains of Escherichia coli K12 and B: differential suppression of its effects on DNA replication, DNA repair and ultraviolet mutagenesis. AB - We have transduced the mutant allele ssb-1, which encodes a temperature-sensitive single-strand DNA binding protein (SSB), into several Escherichia coli strains, and have examined colony-forming ability, DNA replication, sensitivity to ultraviolet light (UV) and UV-induced mutability at the nonpermissive temperature. We have found: 1) that the degree of ssb-1-mediated temperature sensitivity of colony-forming ability and of DNA replication is strain-dependent, resulting in plating efficiencies at 42 degrees C (relative to 30 degrees C) ranging from 100% to 0.002%; 2) that complete suppression of the temperature sensitivity caused by ssb-1 occurs only on nutrient agar, and not in any other medium tested; 3) that strains in which ssb-1-mediated temperature-sensitivity is completely suppressed show moderate UV sensitivity and normal UV mutability at 30 degrees C, but much more extreme UV sensitivity and drastically reduced UV mutability at 42 degrees C; and 4) that defects in excision repair or in other Uvr+-dependent processes are not responsible for most of the UV sensitivity promoted by ssb-1. We discuss our results in relation to the known properties of SSB and its possible role in the induction of DNA damage-inducible (SOS) functions. PMID- 6276688 TI - Site-specific properties of Tn7 transposition into the E. coli chromosome. AB - A study has been made of the insertional properties of transposon Tn7, a 14 kilobase transposable element encoding resistances to trimethoprim, streptomycin and spectinomycin. It has previously been shown that Tn7 transposes at a low frequency and with low specificity into multiple sites in large transmissible plasmids. However, Tn7 transposes with extreme specificity and at high efficiency into the E. coli chromosome. In all cases we have studied, insertion of Tn7 into the chromosome has occurred at a unique site and with a unique orientation. A combination of genetic and biochemical techniques have been used to precisely locate this site on the E. coli chromosome to minute 82 on the linkage map between markers glmS and uncA. To investigate the nature of this highly specific transpositional event, a small region of the E. coli chromosome that includes the unique site, was cloned into the plasmid vector pBR322. Subsequently a lkb restriction fragment, including the Tn7 insertion site, was sub-cloned from this plasmid into the plasmid pACYC184. We show that Tn7 transposes into both these plasmid recombinants with the frequency and specificity characteristic of the E. coli chromosome. PMID- 6276689 TI - DNA sequences at the sites of three insertions of the transposable element Tn5 in the histidine operon of Salmonella. PMID- 6276690 TI - Physical arrangement of suppressor-sensitive mutations of Bacillus phage M2. PMID- 6276691 TI - Molecular cloning of the uvrD gene of Escherichia coli that controls ultraviolet sensitivity and spontaneous mutation frequency. AB - The uvrD gene of Escherichia coli that controls UV sensitivity and spontaneous mutation frequency has been cloned with phage lambda as vector. The increased sensitivity to ultraviolet light (UV) of uvrD3, uvrE502, recL152, and pdeB41 mutants, high mutability of uvrD3 and pdeB41 mutants, and conditional lethality of strain TS41 that carried pdeB41, polA1, and supl26 mutations were all suppressed by lysogenization of the mutant cells with lambda uvrD+. These results were consistent with the idea that the uvrD, uvrE, recL, and pdeB mutations are alleles of the uvrD gene. In addition to the uvrD gene, lambda uvrD+ carried the corA gene that controls transport of Mg++, Mn++, and Co++ through the cell membrane. Hybrid plasmids carrying both uvrD and corA genes were also constructed by using pKY2289 as a cloning vehicle. Orientational isomers that carried the same 12.0 kb fragment in the opposite direction were equally efficient in complementing the UvrD- as well as CorA- defects of the transformed host cells, suggesting that the DNA insert contains all the genetic signals needed to express the two gene products. Insertion of the gamma delta sequence into recombinant plasmids was performed to generate appropriate restriction endonuclease target sites in the cloned DNA fragments. PMID- 6276692 TI - P1 transduction map spanning the replication terminus of Escherichia coli K12. AB - The region of the E. coli chromosome that contains the replication terminus has not previously been spanned by P1 cotransduction. We have used Tn5, Tn9 and Tn10 transposons inserted in this region as genetic markers, and have constructed a genetic map that extends from fnr (min 29.3) to manA (min 35.7). The relevant transposons that have been mapped in this region and which are described in this report are trg-1::Tn5 (min 31.1), zdc-235::Tn10 (min 32.3), zdd-230::Tn9 (min 33.3), and zde-234::Tn10 (min 34.2). The size of this region as determined by P1 cotransduction is very similar to previous estimates obtained by bacterial conjugation. PMID- 6276693 TI - Transposon Tn951 (TnLac) is defective and related to Tn3. AB - Tn951 is flanked by two perfect inverted repeats of 41 bp which include the 38 bp sequence of the IR of Tn3. Tn951 also contains the last 100 bp of the tnpA gene but with at least two mutations. However, beyond nucleotide 137 the sequences diverge and hybridization experiments show that Tn951 lacks at least the first two thirds of the tnpA gene. In agreement with these observations Tn951 does not transpose by itself at a detectable frequency but can be complemented by the tnpA gene of Tn801 or Tn3. Tn501, Tn1721 and gamma delta do not complement Tn951 transposition. Transposition of Tn951 duplicates 5 bp of target DNA sequence. PMID- 6276694 TI - Vectors for cloning in cyanobacteria: construction and characterization of two recombinant plasmids capable of transformation of Escherichia coli K12 and Anacystis nidulans R2. AB - Two plasmids were constructed consisting of the E. coli vector pACYC184 and the cyanobacterial plasmid pUC1. These recombinants, designated pUC104 and pUC105, can be transformed to E. coli K12 as well as to the cyanobacterium Anacystis nidulans R2 and in both hosts they express their antibiotic markers. pUC104 and pUC105 differ with respect to the location and the orientation of the pACYC184 segment in pUC1. pUC104 was found to be stable under all circumstances. Transformation of pUC105 to A. nidulans R2 gave intact plasmids when chloramphenicol was the selective agent, but upon ampicillin selection a deletion derivative was produced identical to pUC1. Further characteristics of pUC104 and pUC105 are described and their usefulness as cloning vectors is discussed. PMID- 6276695 TI - Promoters in the region of the E. coli replication origin. PMID- 6276697 TI - Restriction of streptomyces phage SH5 by endonuclease ShyI from Streptomyces hygroscopicus 0477. PMID- 6276696 TI - Construction and characterization of a tufA-lacZ fusion coding for an E. coli EF Tu-beta-galactosidase chimeric protein. AB - A new phage lambda cloning vector was constructed that has a single EcoRI site upstream from weakly expressed lacI-Z gene isolated by Muller-Hill and Kania (1974). An EcoRI fragment containing the complete tufA gene of E. coli was cloned on the vector and the recombinant phage was crossed into the str operon that has tufA as its last gene. Subsequent selection gave rise to a tufA-lacZ fusion that codes for a chimeric peptide. The fused peptide has a molecular weight of 148,000 and contains 40% of the N-terminal of EF-Tu followed by part of the lac repressor beta-galactosidase fusion. The specific activity of the fused peptide is about half of the activity of normal beta-galactosidase. PMID- 6276699 TI - A new type of IS1-mediated deletion. AB - Genetical tests and DNA sequence analysis revealed that the mechanism of formation of IS1-induced type I and type II deletions differs. IS1-mediated type II deletions occur at the termini of the integrated element and do not remove the element. This process is independent of the cellular recA system and does not involve DNA sequence homology. Conversely, the formation of IS1-induced type I deletions differs substantially. They require recA gene product, small DNA sequence duplications and a topological arrangement of the DNA molecule to allow alignment of duplications. PMID- 6276698 TI - Physical mapping of bacteriophage T4. PMID- 6276700 TI - Increase in microvascular permeability induced by enzymatically generated free radicals. II. Role of superoxide anion radical, hydrogen peroxide, and hydroxyl radical. PMID- 6276701 TI - Comparative activity of N-formimidoyl thienamycin with third generation cephalosporins and ureido penicillins against multiple resistant Serratia marcescens. AB - Twenty multiple resistant clinical isolates were tested with N-formimidoyl thienamycin, moxalactam, cefotaxime, cefoperazone, and the three ureidopenicillins: azlocillin, mezlocillin, and piperacillin. A concentration of less than 0.97 microgram/ml inhibited 100% of organisms for N-f-thienamycin and cefotaxime, 90% for moxalactam, and 60% for cefoperazone. An increase in inoculum from 10(3) to 10(6) cells reduced activity fourfold for 95% of isolates with cefoperazone, 70% with N-formimidoyl thienamycin, 65% for cefotaxime, but only 15% for moxalactam. For ureidopenicillins, 85% of strains tested had MIC's less than or equal to 15.6 micrograms/ml. An inoculum effect was observed in only 35 50%. At 10(3), the cidal concentration was the same or twofold greater than the inhibitory level for N-f-thienamycin and cephalosporins in 70% of strains tested and 65% for penicillins. With 10(6), the 70% value remained for N-f-thienamycin but was reduced to 45% for cefotaxime and 25% for moxalactam; 85% demonstrated greater than eightfold differences with cefoperazone. Single step high-level resistance was observed to moxalactam (20%). Carbenicillin resistant strains were cross-resistant to the ureidopenicillins. N-f-thienamycin and cefotaxime appeared comparable, although important differences between morphological alteration and metabolism may influence their therapeutic effectiveness. PMID- 6276702 TI - Establishment of macrophage-like cell lines of the guinea pig. PMID- 6276704 TI - [Exogenous orthophosphate regulation of the phosphohydrolase activities of Pseudomonas aeruginosa and Pseudomonas maltophilia]. AB - The activity of several phosphohydrolases, viz. alkaline phosphatase (EC 3.1.3.1), acid phosphatase (EC 3.1.3.2), ATPase (EC 3.6.1.3), tripolyphosphatase (EC 3.6.1.2) and polyphosphatase (EC 3.6.1.11), was studied in Pseudomonas aeruginosa VKM B-889 and Pseudomonas maltophilia VKM B-591. In the absence of orthophosphate in the medium when alkaline phosphatase was derepressed, its activity in P. aeruginosa rose in parallel with that of acid phosphatase, tripolyphosphatase and polyphosphatase. The maximal activity of alkaline phosphatase was detected in the cultural broth while that of the remaining enzymes was found in the soluble fraction of the cells. In P. maltophilia, the activity of phosphohydrolases was not regulated with orthophosphate; even when the cells were grown in its presence, the activity was much higher than that of acid phosphatase, ATPase and tripolyphosphatase of the derepressed cells of P. aeruginosa. In P. maltophilia, the maximal activity of the enzymes, just as that of alkaline phosphatase, was detected in the fraction of cellular membranes. PMID- 6276703 TI - [Enzyme activity of aerobic spotulating bacteria isolated from humans and animals]. PMID- 6276706 TI - [Effect of benzidamine, prostacyclin and isoproterenol on the cAMP levels in human lymphocytes]. PMID- 6276707 TI - Feline ophthalmologic diseases. Conjunctival diseases. PMID- 6276708 TI - Halide- and gamma-aminobutyric acid-induced enhancement of diazepam receptors in rat brain. Reversal by disulfonic stilbene blockers of anion channels. PMID- 6276709 TI - gamma-Aminobutyric acid enhancement of CL 218,872 affinity and evidence of benzodiazepine receptor heterogeneity. PMID- 6276710 TI - Structural requirements for specific recognition of mu or delta opiate receptors. PMID- 6276711 TI - Interaction of tricyclic antidepressants with the ionic channel of the acetylcholine receptor of Torpedo electric organ. PMID- 6276705 TI - Cyclic nucleotides in procaryotes. PMID- 6276712 TI - Phenoxybenzamine and dibenamine interactions with calcium channel effectors of the muscarinic receptor. PMID- 6276713 TI - High-affinity saxitoxin receptor sites in vertebrate heart. Evidence for sites associated with autonomic nerve endings. PMID- 6276715 TI - Desensitization of adenylate cyclase to prostaglandin E1 or 2-chloroadenosine. PMID- 6276714 TI - Pharmacological properties of sodium channels in cultured rat heart cells. PMID- 6276716 TI - N alpha-formyl-norleucyl-leucyl-phenylalanyl chloromethylketone. A possible covalent agonist of the N alpha-formyl-methionyl-peptide receptor. AB - N alpha-formyl-norleucyl-leucyl-phenylalanine-chloromethyl ketone is chemotactic for, and induces lysosomal enzyme release from rabbit peritoneal neutrophils over essentially the same range of concentrations as does the free acid form of the same peptide (N alpha-formyl-norleucyl-leucyl-phenylalanine-OH). The chloromethyl ketone derivative does however differ from the free acid in respect to its ability to interact with the neutrophil and cause deactivation or desensitization to cytochalasin B. Neutrophils preincubated in the cold with the chloromethyl ketone followed by washing have cytochalasin B sensitivity conferred upon them, as measured by the release of lysosomal enzymes. The degree of release induced by this pre-treatment appears to be related to the initial responsiveness of the cells. This is in contrast to the free acid where no cytochalasin B sensitivity is conferred under any circumstances. Thus, the chloromethyl ketone, unlike the free acid, appears to irreversibly activate the cell. Desensitization to the late addition of cytochalasin B is also significantly retarded when the chloromethyl ketone derivative is compared to the free acid form of the peptide. These studies suggest that the chloromethyl ketone derivative of the peptide may covalently interact with the neutrophil receptor. PMID- 6276718 TI - 32P-labeling of bovine tryptophanyl-tRNA synthetase with [32P] Pyrophosphate. PMID- 6276717 TI - The serum thymic factor (FTS). Chemical and biological aspects. PMID- 6276720 TI - Periaqueductal gray stimulation in humans produces analgesia accompanied by elevation of beta-endorphin and ACTH in ventricular CSF. PMID- 6276719 TI - Involvement of histone H1 in the structure of the linker DNA in nucleosomes as revealed by nucleases. AB - This report describes experiments designed to study the organization of the linker DNA in nucleosomes. When rat liver nucleosomes (145 to 188 base pairs in length) were digested by Exonuclease III and then by nuclease S1 a series of bands on sizes 90 - 102 - 112 - 125 - 135 - 142 - 154 - 166 - 172 - 181 - bases was observed in denaturing electrophoretic gels. Digestion of H1-depleted nucleosomes under the same conditions results in a series of products of sizes (10.4) xn in base (n less than or equal to 14) only. This result is interpreted as reflecting a particular arrangement of linker DNA under the influence of histone H1. PMID- 6276722 TI - Role of endorphins in addiction. PMID- 6276721 TI - Endorphins in traumatic spinal injury. Pathophysiologic studies and clinical implications. PMID- 6276723 TI - Endorphin hypothesis of schizophrenia. PMID- 6276724 TI - Endorphin studies: electrophysiologic effects. PMID- 6276725 TI - Stress, endorphins and psychosis. A hypothesis. PMID- 6276726 TI - Action of naloxone and naltrexone in different types of psychoses. PMID- 6276727 TI - Clinical studies on the role of endorphins in schizophrenia. PMID- 6276729 TI - Detoxication treatment of schizophrenic patients: a possible role of endorphins? PMID- 6276730 TI - Localization and modulation of enkephalins, endorphins and opiate receptors in the CNS and the pituitary gland. PMID- 6276728 TI - beta-Endorphin studies in psychiatric patients. PMID- 6276731 TI - Stress and endogenous opioid peptides: a review. PMID- 6276732 TI - Role of endorphins in endogenous pain control systems. PMID- 6276733 TI - Pain sensitivity and endorphins in functional psychoses. PMID- 6276734 TI - Arguments for a histogenetic classification of lung carcinomas. AB - Taking into consideration the common origin of lung carcinomas from the basal intermediate cells of covering epithelia of bronchi and bronchioles (which have preserved their ability to divide and differentiate under different stimuli), and the development of tumors in at least two phases, the main histogenetic characteristics of lung carcinomas are discussed. There are pointed out the various and multiple promoting factors acting at the level of proliferating cell foci resulted from the initiating action of chemical carcinogens and the varied deepness of the cellular changes induced. Under these conditions, the malignantly transformed cells may remain poorly differentiated, like during the histogenesis of macrocellular carcinomas (which might be influenced in the course of proliferation, within some limits, by active factors), they may preserve the stereotype of cylindrocubic development, like during the formation of cylindrocubic carcinomas (bronchiolo-alveolar cell carcinomas included), or may suffer metaplastic changes and form subsequently epidermoid carcinomas; the inclusion of Kultschitzky-APUD cells within proliferating foci may lead to bronchial carcinoids or to microcellular carcinomas. The particular forms of lung carcinomas (giant celled, adenocystic, mucoepidermoid) histogenetically reflect the mixed or concomitant stereotypical evolutions of basal cells while preserving some secretory possibilities. PMID- 6276735 TI - Elastosis in breast carcinoma. AB - Elastosis, the significant increase of elastic tissue, was identified by histochemical methods in 45 (53.5%) cases of an unselected series of breast cancers. They were all invasive ductal carcinomas with or without tubular differentiation. The highest proportion of tumors with elastosis was found in the "scirrhous" type of carcinomas. Elastosis was preponderantly of the focal variety, periductal and perivenous. The affected ducts were of large calibre, containing a normal, benign hyperplastic or carcinomatous epithelium. There were not observed correlations with the grade of malignancy and the extent of axillary lymph node metastases. PMID- 6276736 TI - [Endometrial carcinoma in the young woman. Its relations with ovarian lesions]. PMID- 6276737 TI - Effect of BC, 4CMB and 4HMB on the mutation of V79 cells to azaguanine resistance. PMID- 6276738 TI - Evaluation of 4CMB in the subcutaneous implant test. PMID- 6276739 TI - Carcinogenicity bioassay of 4-chloromethylbiphenyl (4CMB), 4 hydroxymethylbiphenyl (4HMB) and benzyl chloride (BC) on mouse skin: interim (7 month) report. AB - 4CMB, 4HMB, BC of benzo[a]pyrene (BP, positive control) dissolved in toluene have been applied twice weekly to the shaved dorsal region of groups of 20 Alderley Park Swiss albino mice. After 7 months of treatment, 6/20 of the BP test group were confirmed as having developed squamous cell carcinoma of the skin. However, none of the animals in the test groups appeared to have been affected by the test chemicals. This interim observation was supplemented by intercurrent sacrifice of 1 animal from each of the test groups, followed by histopathological examination of sections of the subcutaneous tissue. No neoplastic changes were discerned. PMID- 6276740 TI - Attempts to initiate skin tumours in mice in the 2-stage system using 4 chloromethylbiphenyl (4CMB), 4-hydroxymethylbiphenyl (4HMB), and benzyl chloride (BC). Report of the experiment at 10 months. AB - Groups of T.O. mice (Theiler's Original, derived from Swiss albino mice) were treated topically, with a single dose of 1.0 mg of 4CMB, 4HMB, or BC, or with 0.4 mg benzo[a]pyrene (B[a]P). Twice-weekly promotion of the treated skin area with croton oil was begun 1 week later. At 10 months the skin-tumour incidence in the positive control (B[a]P) was 8/19, with a mean latent period of 20 weeks. Both 4CMB and 4HMB have so far produced 1 papilloma each (1/36 at 40 weeks, and 1/39 at 34 weeks, respectively), while BC has produced none. Further time is required in order to ascertain whether these single papillomas will develop into carcinomas and thereby herald weak initiating activity for 4CMB and 4HMB. PMID- 6276742 TI - Activated oxygen species at the origin of chromosome breakage and sister chromatid exchanges. PMID- 6276741 TI - The UKEMS genotoxicity trial. A summary of the assays for skin tumour induction in mice, the subcutaneous implant test and the sebaceous gland suppression test. PMID- 6276743 TI - Inhibition of DNA synthesis in relation to enhanced survival of UV-damaged herpes virus in monkey cells treated by a variety of 2-nitronaphthofurans. AB - Various 2-nitronaphthofuran derivatives (related to each other by simple structural modifications) were tested for 2 different effects in CV-1 monkey kidney cell cultures: the immediate inhibition of normal DNA synthesis and the capacity of pretreated cultures (40 h of contact) to support the replication of UV-damaged Herpes simplex virus (HSV). For all compounds tested, a fair correlation was found between their efficiencies to inhibit cellular DNA synthesis and to provoke an increase in UV-HSV production (virus reactivation). Virus reactivation was due to an increase in both the number of virus-producing cells and the amount of infectious particles produced per cell. The most efficient 2-nitronaphthofurans (particularly 2-nitro-7-methoxy-naphtho[2,1 b]furan-R 7000) were at least as potent as aflatoxin B1 in inducing virus reactivation. PMID- 6276744 TI - Protonmotive force in muscle mitochondria. AB - The protonmotive force (delta p) of muscle mitochondria was measured by estimating the distribution of 14C-labeled TPMP (trimethylphenylphosphonium iodide) and 14C-labeled acetate across the inner membrane of muscle mitochondria. The matrix volume was simultaneously determined using 3H-labeled H2O and 3H labeled mannitol and repeated drying to distinguish the label in these 2 compounds. Rapid separation of mitochondria from the incubation medium by centrifugation through silicone oil avoids the problems of potential anaerobic conditions associated with conventional centrifugation and large volumes of trapped media associated with filtration. The value for delta p (mean +/- SD) was 192+/- 26 mV in 30 determinations with rat muscle mitochondria during state 4. Measurement of oxygen consumption allowed calculation of membrane conductance (Cm,H+) which was 0.49 +/- 0.18 nmol of H+/min/mg protein/mV. The values for delta p and Cm,H+ are reported for a variety of experimental conditions and are consistent with Mitchell's chemiosmotic theory. Biopsy specimens obtained from human muscle gave state-4 delta p values of 197+/- 30 mV (n =5) and Cm,H+ values of 0.52 +/- 0.12 nmol of H+/min/mg/mV (n = 4). This delta p assay is the first described for coupled mammalian muscle mitochondria and will be useful in assessing membrane function. PMID- 6276745 TI - Defect in ACTH control after hypophysectomy in Cushing's disease. PMID- 6276746 TI - Effect of the fluoride/calcium regimen on vertebral fracture occurrence in postmenopausal osteoporosis. Comparison with conventional therapy. AB - We assessed the rates of vertebral fracture in patients with postmenopausal osteoporosis. Forty-five patients were not treated (91 person-years of observation); 59 were treated conventionally, with calcium (alone or combined with estrogen) or vitamin D or both (218 years); and 61 were treated with sodium fluoride combined with conventional therapy (251 years). The fracture rate (per thousand person-years) was 834 in untreated patients, 419 in those given calcium with or without vitamin D, 304 in those given fluoride and calcium with or without vitamin D, 181 in those given estrogen and calcium with or without vitamin D, and 53 in those given fluoride, estrogen, and calcium with or without vitamin D. It was reduced in all treatment groups (P less than 0.001 for calcium and P less than 1 x 10(-6) for other combinations); fluoride (one years of treatment) and estrogen (but not vitamin D) independently reduced the rate from that observed with calcium alone (P less than 0.001). The combination of calcium fluoride, and estrogen was more effective than any other combination (P less than 0.001). These results provide grounds for optimism about the efficacy of combinations of available agents with sodium fluoride for fracture in postmenopausal osteoporosis. PMID- 6276747 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 8-1982. Diffuse pulmonary infiltration in a patient with lymphoma. PMID- 6276748 TI - Virus-induced diabetes in mice prevented by a live attenuated vaccine. PMID- 6276749 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 9-1982. Atypical white cells in the blood of an elderly woman. PMID- 6276750 TI - Cellular origin and role of mink cell focus-forming viruses in murine thymic lymphomas. PMID- 6276751 TI - Three classes of mouse H-2 messenger RNA distinguished by analysis of cDNA clones. PMID- 6276753 TI - DNA sequences necessary for transcription of the rabbit beta-globin gene in vivo. AB - The DNA sequences required for the expression of the rabbit-beta-globin gene in vivo have been examined. A variety of mutant rabbit beta-globin gene templates were linked to a simian virus 40-plasmid recombinant and introduced into HeLa cells; in these conditions the rabbit beta-globin gene is expressed from its own promoter. Comparison of the level of beta-globin transcripts in a variety of deletion mutants shows that for efficient transcription, both the ATA or Goldberg Hogness box, and a region between 100 and 58 base pairs in front of the site at which transcription is initiated, are required. Deletion of either of these regions results in a decrease in the level of beta-globin transcripts by an order of magnitude; deletion of the ATA box causes an additional loss in the specificity of the site of initiation of RNA synthesis. The DNA sequences downstream from the ATA box, including the natural beta-globin mRNA cap site, are dispensable for transcription in vivo. PMID- 6276752 TI - Polyoma virus capsid structure at 22.5 A resolution. AB - X-ray diffraction data from polyoma capsid crystals were phased by refinement of low-resolution starting models to obtain a self-consistent structural solution. The unexpected result that the hexavalent morphological unit is a pentamer shows that specificity of bonding is not conserved among the protein subunits in the icosahedrally symmetric capsid. PMID- 6276754 TI - Hypothalamic paraventricular nucleus is a privileged site for brain-pituitary interaction in long-term tissue culture. PMID- 6276755 TI - New Epstein-Barr virus variants from cellular subclones of P3J-HR-1 Burkitt lymphoma. PMID- 6276756 TI - Evidence from mtDNA sequences that common laboratory strains of inbred mice are descended from a single female. PMID- 6276757 TI - H-2-like genes in the Tla region of mouse chromosome 17. PMID- 6276758 TI - A problem for the theory of biological structure. PMID- 6276759 TI - Cloning and sequence analysis of cDNA for bovine adrenal preproenkephalin. AB - The nucleotide sequence of cloned cDNA for preproenkephalin from bovine adrenal medulla indicates that the precursor protein contains four copies of Met enkephalin and one copy each of Leu-enkephalin, Met-enkephalin-Arg6-Phe7 and Met enkephalin-Arg6-Gly7-Leu8, a previously undetected opioid peptide. The enkephalin and extended enkephalin sequences are each bounded by paired basic amino acid residues. Preproenkephalin may represent a multi-hormone precursor, like the corticotropin-beta-lipotropin precursor. PMID- 6276760 TI - Multiple arrangements of viral DNA and an activated host oncogene in bursal lymphomas. AB - Proviruses of avian leukosis virus (ALV) are located in the vicinity of a putative cellular oncogene (c-myc) in ALV-induced bursal lymphomas. Enhanced expression of c-myc occurs in association with proviruses found in any of three configurations: (I) on the 5' side ('upstream') of c-myc in the same transcriptional orientation; (II) on the 3' side ('downstream') of c-myc in the same orientation; (III) upstream, in the transcriptional orientation opposite to that of c-myc. Thus, activation of adjacent cellular genes by retroviral DNA can involve mechanisms other than provision of a transcriptional promoter. PMID- 6276761 TI - Rous sarcoma virus-induced phosphorylation of a 50,000-molecular weight cellular protein. PMID- 6276763 TI - Altered methylation of endogenous viral promoter sequences during mammary carcinogenesis. PMID- 6276762 TI - Endogenous MuLV infection does not contribute to onset of radiation- or carcinogen-induced murine thymoma. PMID- 6276764 TI - High frequency of gene transfer after fusion between bacteria and eukaryotic cells. PMID- 6276765 TI - Is phosphatidylinositol now out of the calcium gate? PMID- 6276766 TI - Nucleotide sequence of cloned cDNA encoding bovine arginine vasopressin neurophysin II precursor. AB - The sequence of a cDNA encoding the nonapeptide arginine vasopressin (AVP) and its carrier protein, neurophysin II (NpII) from bovine hypothalamus, proves that the 166-amino acid precursor molecule contains a signal peptide of 19 amino acids followed directly by AVP connected to NpII by a Gly-Lys-Arg sequence. The carboxy terminal region of the precursor contains a naturally occurring glycopolypeptide of 39 amino acids which is separated from NpII by a single arginine residue. PMID- 6276767 TI - Excision of polyoma virus genomes from chromosomal DNA by homologous recombination. PMID- 6276768 TI - Rapid kinetics of single glutamate-receptor channels. PMID- 6276769 TI - Isolation from murine sarcoma cells of novel transforming growth factors potentiated by EGF. PMID- 6276770 TI - A hypothesis for the initiation of genetic recombination in eukaryotes. PMID- 6276771 TI - Unusual interactions of benzodiazepine receptor antagonists. PMID- 6276772 TI - Molecular size of benzodiazepine receptor in rat brain in situ: evidence for a functional dimer? PMID- 6276773 TI - Crystal structure of a polyfunctional macrocyclic K+ complex provides a solid state model of a K+ channel. PMID- 6276774 TI - TRNA precursor transcribed from a mutant human gene inserted into a SV40 vector is processed incorrectly. PMID- 6276775 TI - Activation of SV40 genome by 72-base pair tandem repeats of Moloney sarcoma virus. AB - The simian virus 40 (SV40) 72-base pair (bp) tandem-repeated sequences have a crucial role as an activator element in viral gene expression. We replaced the SV40 72-bp repeat with a 72-bp repeat derived from the long terminal repeat (LTR) of cloned Moloney murine sarcoma virus (MSV) DNA. Although there is no detectable sequence homology to SV40, the MSV repeats can substitute functionally for the SV40 repeats and generate a viable virus in monkey kidney cells. PMID- 6276776 TI - Identification of a beta-endorphin-like peptide in cultured human placental cells. PMID- 6276777 TI - Similar mode of action of pyrethroids and DDT on sodium channel gating in myelinated nerves. PMID- 6276778 TI - Long terminal repeats of endogenous mouse mammary tumour virus contain a long open reading frame which extends into adjacent sequences. PMID- 6276779 TI - Inter-replicon transposition of Tn1/3 occurs in two sequential genetically separable steps. PMID- 6276780 TI - Non-dopaminergic fibres may regulate dopamine-sensitive adenylate cyclase in the prefrontal cortex and nucleus accumbens. PMID- 6276781 TI - Cyclic AMP induces maturation of trout sperm axoneme to initiate motility. PMID- 6276782 TI - Homocopolymer sequences in the spacer of a sea urchin histone gene repeat are sensitive to S1 nuclease. PMID- 6276783 TI - K+ depletion and Na+ pump density. PMID- 6276784 TI - Peptides and the new endocrinology. AB - The discovery of regulatory peptides common to the nervous and the endocrine systems (brain, gut, and skin) has brought about a revolution in our concepts of endocrinology and neurology. We are beginning to understand some of the complex interrelationships between soma and psyche that might, someday, be important for an integrated treatment of diseases. Examples of the actions of certain peptides in the periphery and in the central nervous system are given, and their biosynthesis and molecular anatomy as carriers for information are discussed. PMID- 6276785 TI - [125I]-HEAT, a selective, high-affinity, high specific activity ligand for alpha 1-adrenoceptors. PMID- 6276786 TI - Effects of piracetam on the central dopaminergic transmission. AB - The effects of piracetam on dopamine metabolism in rat striatal tissue and on behavioural and biochemical effects of haloperidol were studied. Piracetam in doses 600 mg/kg and 1,000 mg/kg (i.p.) increased the levels of dopamine metabolites (homovanillic acid and 3,4-dihydroxy-phenylacetic acid) but did not influence the content of dopamine in the striatum. Piracetam (1,000 mg/kg) also increased haloperidol-induced catalepsy in rats. These results are discussed in relation to the hypothesis that some behavioural effects of piracetam can be connected with its action on dopaminergic transmission. PMID- 6276787 TI - Involvement of cyclic AMP in the direct inotropic action of amrinone. Biochemical and functional evidence. PMID- 6276788 TI - The effects of p-chloromercuriphenylsulfonic acid (PCMBS) on force of contraction of mammalian myocardium and on ATP hydrolysis by sarcolemmal ATPase. PMID- 6276789 TI - Thromboxane synthetase inhibitors differentially antagonize thromboxane receptors in vascular smooth muscle. AB - A number of thromboxane (Tx) synthetase inhibitors have been found to prevent thromboxane release in acute cardiopulmonary disorders. However, little is known about Tx receptor antagonism by these substances. Imidazole, UK-37,248 and pinane thromboxane A2 (PTA2) were tested in isolated perfused cat coronary arteries, spirally cut rabbit pulmonary artery strips, and in rabbit and cat platelets for their ability to antagonize vasoconstrictor and aggregatory effects of a stable Tx agonist, carbocyclic thromboxane A2 (CTA2). Imidazole, at concentrations that completely inhibit thromboxane synthesis, failed to antagonize the vasoconstrictor effects of CTA2 in both systems. UK-37,248, at 10 to 1000 micrograms/ml, failed to inhibit CTA2-induced coronary constriction but at 1 microgram/ml reduced rabbit pulmonary artery constriction by 80 +/- 8% (P less than 0.0005). In comparison, 1 micro M PTA2 completely prevented PTA2-induced constriction in both coronary and pulmonary arteries. PTA2 did not antagonize KCl and angiotensin II responses. In platelets, PTA2 but not UK-37,248 prevented arachidonate induced aggregation in rabbit and cat platelets. Some of the beneficial effects of thromboxane synthetase inhibitors may be associated with thromboxane receptor antagonism. Inhibitors structurally similar to TxA2 appear to have greater thromboxane receptor antagonistic activity. This additional activity may be of importance in therapeutics of coronary and pulmonary disorders. PMID- 6276790 TI - Effects of drugs affecting the noradrenergic system on convulsions in the quaking mouse. AB - Handling-induced convulsions in the quaking mouse can be blocked by: phenobarbital, pentobarbital or phenytoin; postsynaptic alpha-adrenoceptor agonists (noradrenaline, phenylephrine, CRL 40028); presynaptic alpha adrenoceptor blockers (yohimbine, mianserine); catecholamine liberating agent (amphetamine); noradrenaline reuptake inhibitors (cocaine, imipramine, desipramine). Moreover, the protective effect of yohimbine was antagonized by clonidine, prazosin or alpha-methylparatyrosine, and the protective effect of CRL 40028 was antagonized by prazosin but not by alpha-methyltyrosine. Drugs acting by other mechanisms (pilocarpine, atropine, trihexyphenidyl, (--)-5-HTP, methysergide, pimozide, clonidine, alpha-methyl DOPA, prazosin, isoprenaline, salbutamol) did not protect against convulsions. A slight protection was obtained with high doses of apomorphine and also with (+/-)-propranolol. This effect is probably not related to blockade of beta-adrenoceptors because the same effect was obtained with (+)propranolol. In young quaking mice, where susceptibility to convulsions is low, both postsynaptic alpha-adrenoceptor blockers and presynaptic alpha-adrenoceptor antagonist lowered the convulsive threshold. Thus, this seems to constitute an interesting model for the in vivo study of substances which affect the central alpha-adrenoceptors either pre- or postsynaptically. PMID- 6276791 TI - [Excision of a chemodectoma of the glomus intravagale after preceding embolization; advantages of the combined method]. PMID- 6276792 TI - [A caput Medusae which disappeared again]. PMID- 6276793 TI - [Backache in patients with cancer]. PMID- 6276794 TI - [Regulation of the contraction condition of the venous system of the legs]. PMID- 6276796 TI - [Naloxone-dependent morphine-induced depression of the snail response to serotonin]. AB - Morphine which was added to the extracellular solution in a concentration of 1.10(-5) M, rapidly and reversibly depressed depolarizing (15 cells of 20) and hyperpolarizing (6 cells of 12) serotonin (1.10(-6) M) responses of neurons of the mollusc Helix lucorum. The depressing morphine effect was antagonized by naloxone (1.10(-5) M). The results suggest involvement of an opiate receptor mechanism. Dose-response curves for serotonin before morphine application and in the presence of morphine show a noncompetitive mechanism of morphine-serotonin interaction. PMID- 6276795 TI - [Cortical control of afferent flow in the lateral geniculate body in the cat]. AB - In experiments carried out on cats immobilized with d-tubocurarine unit responses within the dorsolateral geniculate nucleus were analyzed following visual cortex (testing) and optic chiasm (conditioning) stimulation. Two types of responses were found indicating the existence of principal and inhibitory cells in the nucleus. Optic chiasm stimulation was found to produce an inhibitory corticofugal effect on the responses of principal cells. A conclusion is made that these effects are mediated by inhibitory cells. PMID- 6276797 TI - [Synaptic excitation of reticulospinal neurons during the startle reaction in cats anesthetized with chloralose]. AB - Synaptic processes in the bulbar and pontine reticulospinal neurons (RS-neurons) were studied in chloralose-anesthetized cats during startle reaction to somatic stimulation ("spino-bulbo-spinal"--SBS-reflex). Excitatory PSPs were found to be a main type of synaptic responses in the RS-neurons; they were oligosynaptic (including monosynaptic) and polysynaptic. The comparison between synaptic responses of RS-neurons and SBS-reflexes showed that the polysynaptic EPSPs were most effective in generation of a SBS descending volley. It was also shown that about half of bulbar and pontine RS-neurons were simultaneously involved in suprasegmental relay of the SBS-reflexes. Conduction velocities in their axons ranged between 30 and 98 m/s (mean 64.5 +/- 16.5 m/s). Some functional features of the reticular centre of startle reaction are discussed. PMID- 6276798 TI - [Effect of theophylline on the electrical activity of the Helix pomatia neuron PPa2]. PMID- 6276799 TI - [Effects of a psychiatric hospital outpatient service on the problem of mental health care (author's transl)]. PMID- 6276800 TI - Prostaglandins and leukotrienes in inflammation and allergy. PMID- 6276801 TI - [Glomerular nephropathies and viral infections]. PMID- 6276802 TI - Influence of adrenocorticotropin and adrenalectomy on gonadotropin secretion in immature rats. AB - We examined the effects and mechanisms of action of ACTH and ACTH fragments on gonadotropin secretion in immature rats. ACTH administered by daily injection or continuous infusion (osmotic minipumps) attenuated the postcastration rise in serum LH. Pituitary LH concentration was either unchanged or increased in ACTH treated rats and pituitary sensitivity to gonadotropin-releasing hormone (GnRH) was reduced by ACTH treatment. A fragment of ACTH (ACTH 4-10), which is less steroidogenic, did not alter levels of serum LH, and ACTH did not reduce LH secretion in adrenalectomized castrates. Serum and pituitary concentrations of prolactin were normal in ACTH-treated animals. These studies demonstrate that the suppression of gonadotropin secretion by ACTH is mediated by the adrenal gland. This mechanism causes a decreased pituitary sensitivity to GnRH, but LH synthesis does not appear to be affected. Prolactin does not play a role in this mechanism. PMID- 6276803 TI - Evidence that the effects of 5-hydroxytryptophan on the secretion of ACTH and growth hormone in dogs are not mediated by central release of serotonin. AB - 5-Hydroxytryptophan (5-HTP) was found to lower plasma 11-oxycorticoids and increase plasma growth hormone in anesthetized dogs. To determine whether these effects were mediated by release of serotonin, 5-HTP and L-tryptophan were injected intravenously alone and after treatment with various drugs. The decrease in corticoids produced by 5-HTP was unaffected by intravenous carbidopa and metergoline, but was abolished by intravenous benserazide in a dose that inhibits central 5-HTP and dopa decarboxylase, and by a small dose of phenoxybenzamine in the third ventricle. Adrenal sensitivity to ACTH was unaffected by 5-HTP. L Tryptophan did not decrease plasma corticoids; instead it produced a small increase. The increase in plasma growth hormone produced by 5-HTP was potentiated by carbidopa and abolished by benserazide. L-Tryptophan did not increase plasma growth hormone; instead it produced no change or in some instances a decrease. Plasma growth hormone declined after administration of phenoxybenzamine in the third ventricle, and the decline continued after administration of 5-HTP. The data indicate that the effects of 5-HTP on ACTH and growth hormone secretion are not due to central release of serotonin and suggest that they are due instead to serotonin-induced release of catecholamines from catecholamine-secreting neurons. PMID- 6276804 TI - Patterns of cyclic AMP phosphodiesterases in the rat pineal gland: sex differences in diurnal rhythmicity. AB - Rat pineal cyclic AMP (cAMP) phosphodiesterases (PDEs) were studied during male development and in adult females by means of gel electrophoresis, kinetic analysis, and diurnal activity measurements. In both male and female rats, two molecular forms of PDE coexisted. In neonatal-infantile males (birth to day 20 of age) a third isozyme was detected. The kinetic properties of PDEs were essentially similar in infantile and adult male rats with respect to the Kms; however, a minimal Vmax was observed at 10.00 a.m. in adults, but not in neonatal animals. PDE activity in male rats was maximal between 2.00 a.m. and 4.00 a.m., and also at 4.00 p.m., and was minimal at 10.00 a.m. This pattern appeared to develop during the infantile period of life. The pattern of activity observed in adult female rats was similar to that of adult males, except that it lacked a pronounced afternoon peak. The diurnal PDE activity profile of castrated male adult rats was similar to female adults and testosterone substitution in male castrated rats resulted in a pattern which resembled that of adult male controls. In addition to the above measurements, testosterone serum levels have been determined during all of these experiments. PMID- 6276805 TI - Thalamic-hypothalamic interrelationships and stress-induced rebounding adrenocortical response in the pigeon. AB - Plasma corticosterone levels were determined by protein-binding assay at 2- and 5 min intervals after systemic or neurogenic stress. The same corticosterone (B) profile was observed after hemispherectomy as in intact pigeons, exhibiting episodic increase including several successive peaks after stress application. Hypothalamic stimulation of ACTH injection resulted in a simpler adrenocortical response, analogous to the first peak of the response to stress. Postero-lateral deafferentation of the hypothalamus unabled neurogenic stress to promote any plasma B increase and significantly altered the response to systemic stress since only the first peak of B occurred. Such a single peak response to both systemic and neurogenic stresses was observed after surgical interruption of neural connections between hypothalamic and thalamic structures without interferring with posterior afferents to the basal hypothalamus. The role of thalamic hypothalamic interrelationships in modulating the stress-induced adrenocorticotropic activity is discussed. PMID- 6276806 TI - Lack of benefit from magnesium in lithium toxicity. PMID- 6276807 TI - Effect of isolated and combined stimulation of thalamic nuclei on the character of functional connections between sensomotor cortical neurons. PMID- 6276809 TI - The response of rat spinal cord cells to unmyelinated afferents after peripheral nerve section and after changes in substance P levels. PMID- 6276808 TI - Separation of potassium and calcium channels in the nerve cell soma membranes. AB - Investigation of isolated neurons of Helix pomatia during intracellular dialysis revealed differences in the sensitivity of the channels for the outward potassium and inward calcium currents to changes in pH of the external medium. As a result of this difference, considerable separation of the regions of activation of the currents was obtained along the potential axis in solutions with low pH and the characteristics of the inward and outward currents could be studied during their minimal application. Channels for the outward current were shown to have some permeability for tris ions (PTris : PK=0.05), which is the reason why it is impossible to block this current completely by replacing the intracellular potassium by Tris. Channels for the inward calcium current are characterized by slow inactivation, with a first-order kinetics; their momentary voltage-current characteristic curve reveals significant Goldman's rectification. The selectivity of the calcium channels for other bivalent cations is : Ba:Sr:Ca:Mg=2.8:2.6:1.0:0.2. PMID- 6276810 TI - A study of the cutaneous afferent input to substantia gelatinosa. PMID- 6276811 TI - Diamide, a thiol oxidizing agent, uncouples the electrotonic junctions of crayfish septate axons, but not those of Navanax motoneurons. PMID- 6276812 TI - Interaction of DDT with sodium channels in squid giant axon membranes. PMID- 6276813 TI - Potassium-induced secretion of melanocyte-stimulating hormone from isolated pars intermedia cells signals participation of voltage-dependent calcium channels in stimulus-secretion coupling. PMID- 6276814 TI - Release of endogenous 5-hydroxytryptamine from resting and stimulated enteric neurons. PMID- 6276815 TI - Depression of nigral pars compacta cell discharge by exogenous acetylcholinesterase. PMID- 6276816 TI - [Instrumental diagnosis of deep venous thrombosis. Reliability of phleboscintigraphy and the Doppler method as compared with phlebography]. PMID- 6276817 TI - [Occult insulinoma. New prospects for the therapeutic approach]. PMID- 6276818 TI - [Rare case of chemodectoma with a rare mediastinal location]. PMID- 6276819 TI - [The importance of intestinal pH in the appearance of colo-rectal tumors]. PMID- 6276820 TI - Autoradiographic localization of opioid receptors in the mammalian retina. PMID- 6276821 TI - Hypothalamic deafferentation in the rat appears to discriminate between the anterior lobe and intermediate lobe response to stress. AB - Anterolateral deafferentation of the mediobasal hypothalamus prevented the increase of the plasma corticosterone concentration induced by ether, histamine and electric footshock. Hypothalamus deafferentation also prevented the ether stress-induced increase of the plasma levels of ACTH and beta-endorphin immunoreactivity (ACTHi, beta-ENDi). Infusion of isoproterenol evoked an increase of the plasma levels of corticosterone, ACTHi, beta-ENDi and alpha-MSH immunoreactivity (alpha-MSHi) in sham-operated rats. In rats with a deafferented hypothalamus, the responses of plasma corticosterone and ACTHi to isoproterenol were blocked but the responses of plasma beta-ENDi and alpha-MSHi remained intact. We conclude that circulating beta-ENDi after exposure to ether is of anterior lobe origin while circulating beta-ENDi after infusion of isoproterenol is of intermediate lobe origin. PMID- 6276822 TI - Upper motor neurone modulation of charge movement and mechanical activation in rat skeletal muscle fibres. PMID- 6276823 TI - Upper motor neurone modulation of the structure of the terminal cisternae in rat skeletal muscle fibres. AB - There are fewer indentations on the flat surfaces of terminal cisternae in soleus (slow-twitch) than in extensor digitorum longus (EDL, fast-twitch) muscle fibres of rats. Following mid-thoracic spinal cord transection, there is an increase in the number of indentations in soleus fibres but no change in EDL fibres. The increase in the numbers of indentations after spinal cord transections is correlated with changes in the contractile and charge movement properties of the soleus fibres so that they resemble normal EDL fibres. The indentations appear to have an important role in excitation-contraction coupling. PMID- 6276824 TI - Facilitation of transmission in Ib pathways by cutaneous afferents from the contralateral foot sole in man. AB - Changes in the quadriceps H-reflex were used to study the effect of a weak contralateral cutaneous stimulation upon transmission in Ib pathways from ankle muscles to quadriceps in man. Provided it was applied to the contralateral foot sole, such a stimulation facilitated transmission in Ib pathways to quadriceps (inhibitory from extensors as well as excitatory from flexors). The central latency of this contralateral cutaneous facilitation was 1 msec longer than that of the ipsilateral cutaneous depression of Ib pathways. These findings are discussed with regard to the requirements of bipedal gait. PMID- 6276825 TI - Are endogenous opiates involved in potentiation of field potentials in the hippocampus of the rat? AB - Changes in hippocampal CA1 field potentials during and after tetanic stimulation were studied in rats pretreated with 5 mg/kg s.c. naloxone or saline. Stimuli consisted of successive tetani of increasing frequency; responses at threshold, midrange and asymptotic intensities were sampled. A several fold increase in response amplitude occurred during low frequency stimulation, with no significant differences between the groups. Post-tetanic responses showed depression at the lower frequencies but increases ranging up to 400% at higher frequencies, with the greatest potentiation at threshold; again there were no significant differences between naloxone and saline groups, suggesting that endogenous opiates are not involved. PMID- 6276826 TI - Evidence for analgesic activity of beta-casomorphin in rats. PMID- 6276827 TI - On the interactive role of central noradrenaline neurons and corticosterone in two-way active avoidance acquisition in the rat. AB - The noradrenaline (NA) neurotoxin, DSP4, caused a marked impairment of two-way active avoidance acquisition. Pretreatment with desipramine, which inhibits the degeneration of NA neurons by DSP4, consistently blocked the avoidance deficit. Daily treatment with corticosterone (2 X 1 mg/kg, s.c.) also blocked the impaired acquisition of avoidance induced by DSP4 but failed to affect the increase in cortical beta-noradrenergic receptors induced by DSP4. The present findings give further evidence for the important role interactions between the pituitary adrenal axis and the locus coeruleus NA system play in aversive learning. PMID- 6276828 TI - Depolarization-induced release of folates from slices of rat cerebellum. PMID- 6276829 TI - Beta-adrenergic receptors in rat cerebellum after neonatal X-irradiation: effect of prolonged imipramine and lithium treatment. AB - The actions of prolonged treatment of rats within imipramine or lithium chloride on the beta-adrenergic cAMP generating system of the cerebellum were compared in normal animals and in rats degranulated by neonatal X-irradiation. Whereas in normal animals imipramine treatment is without significant effect on the cerebellar cAMP system but lowers the beta-adrenoceptor density significantly, in degranulated animals the cAMP system becomes subsensitive towards norepinephrine after prolonged imipramine treatment. These data support the hypothesis that in the cerebellum imipramine acts predominantly on beta-adrenoceptors located particularly on Purkinje cells. Prolonged lithium treatment exhibited no significant effects on beta-receptors nor the noradrenergic cAMP generating system of normal and degranulated cerebella. PMID- 6276830 TI - Nonanemic pernicious anemia. Delay in diagnosis. PMID- 6276831 TI - Lumpectomy and chemotherapy. Unproved method of treating breast cancer. PMID- 6276832 TI - Metastatic bronchogenic carcinoma; endoscopically proved remission. PMID- 6276833 TI - Long-term survival determinants in Wilms' tumor. AB - This is a retrospective report of 54 patients with the diagnosis of Wilms' tumor treated at Roswell Park Memorial Institute from 1946 to 1977. The age ranged from 1 month to 57 years with a mean of 7.2 years and a median age of 5 years. An analysis of the survival results showed that patients continued to die of disease until 66 months post-diagnosis, making the 5-year survival a much better evaluation criterion for this group than the traditional 2-year survival. Males fared significantly better in this series (p less than 0.01). This was not due to detectable differences in age group, or histology. While the older age group (more than 15 years old) showed a somewhat better survival outlook for the first 2 years post-diagnosis, this difference became negligible at 5 years post diagnosis. PMID- 6276834 TI - Immunohistological demonstration of lysozyme in human leucocytes, salivary corpuscles and nasal discharge cells after blockade of myeloperoxidase activity. PMID- 6276835 TI - The immunology of cancer of the head and neck with particular reference to oral cancer. PMID- 6276836 TI - Rates of fixation by lightning of carbon and nitrogen in possible primitive atmospheres. AB - A thermochemical-hydrodynamic model of the production of trace species by electrical discharges has been used to estimate the rates of fixation of C and N by lightning in the primitive atmosphere. Calculations for various possible mixtures of CH4, CO2, CO, N2, H2, and H2O reveal that the prime species produced were probably HCN and NO and that the key parameter determining the rates of fixation was the ratio of C atoms to O atoms in the atmosphere. Atmospheres with C more abundant than O have large HCN fixation rates, in excess of 10(17) molecules J-1, but small NO yields. However, when O is more abundant than C, the NO fixation rate approaches 10(17) molecules J-1 while the HCN yield is small. The implications for the evolution of life are discussed. PMID- 6276837 TI - The radiolysis and radioracemization of amino acids on silica surfaces. AB - L-Leucine, deposited on both 1-quartz powder and on a commercial amorphous silica preparation (Syloid 63), has been subjected to irradiation in a 60Co gamma-ray source, and the ensuing radiolysis and radioracemization have been determined gas chromatographically. The radiolysis and radioracemization observed for leucine on 1-quartz were rather similar to those noted for a crystalline L-leucine control. L-Leucine on Syloid 63, however, was vastly more susceptible to radiolysis as compared to the L-leucine control, and radioracemization was also markedly enhanced - each increasing with larger radiation dosage. L-Isovaline showed a similar, but diminished, enhancement of radiolysis sensitivity when adsorbed on the Syloid surface, but underwent no radioracemization whatsoever. The divergent results of the control and quartz-leucine irradiations versus the Syloid-leucine and Syloid-isovaline irradiations are interpreted qualitatively in terms of the surface area parameters of the two silica adsorbents and the amino acid adsorbates. PMID- 6276838 TI - Search for effect of longitudinally polarized protons on optically active amino acid. AB - As a test of the Vester-Ulbricht hypothesis that the specific optical activity of contemporary biology arose from a differential action of polarized nuclear particles, DL-leucine has been irradiated with protons of both positive and negative longitudinal polarization. Neither kind of protons caused by measurable decomposition difference in the two optical isomers of the amino acid. PMID- 6276839 TI - [Male breast cancer associated with bilateral Leydig cell hyperplasia]. PMID- 6276840 TI - Diagnosis and treatment of allograft rejection. AB - Several immunologic tests hve been developed to predict allograft rejection prior to functional renal impairment. Unfortunately, no test has as yet proven unequivocally to be of clinical value. Serial serum creatinine determinations and 131I hippuran scintiphotography have proven to be the most useful tests in detecting rejection episodes. Treatment of an acute rejection episode is often successful, but chronic rejection causes a slowly progressive deterioration of allograft function and is unresponsive to treatment. PMID- 6276841 TI - [Generalized cytomegalic inclusion disease (author's transl)]. PMID- 6276842 TI - Spinal opiate analgesia: characteristics and principles of action. PMID- 6276843 TI - Non-opioid pathways suppress pain in humans. AB - Subcutaneous electrical stimulation (SCNS) of median, radial, and saphenous nerves produces prolonged analgesia. The effect of subcutaneous nerve stimulation (SCNS) is not mediated by opiate receptors. (1) There is no cross-tolerance with opiates, and SCNS suppresses pain in patients under chronic meperidine medication. (2) Patients unresponsive to morphine benefit from SCNS. (3) There is no development of tolerance to SCNS. (4) SCNS analgesia is not naloxone reversible. (5) SCNS can be administered with oral analgesics, either opiate or non-opiate, increasing patient comfort. These observations suggest that non endorphinergic pathways produce powerful analgesia. PMID- 6276844 TI - [Prostaglandins and hypertension]. PMID- 6276846 TI - Pathogenesis of gestational trophoblastic neoplasms. PMID- 6276845 TI - Pathobiology of hepatitis B virus. PMID- 6276847 TI - Parathyroid hormone: chemistry and structure-activity relations. AB - Peptide synthesis and the application of a wide range of biological assays have permitted intensive and detailed study of structure-activity relations for parathyroid hormone. Within the structure of the hormone molecule reside largely distinct domains critical for receptor binding or activation of adenylate cyclase in addition to receptor binding. Subtle modifications of hormonal structure can cause striking changes in hormone potency or in the nature of the biological properties displayed by such analogs. For parathyroid hormone, structure-activity studies have identified several discrete regions of the molecule that are responsible for independent biological functions. It was determined that these separate functions are displayed in an almost linear fashion along the primary sequence of the hormone--a conceptual framework that has greatly facilitated synthesis of parathyroid hormone analogs. The amino-terminal region of the initially biosynthesized precursor form of parathyroid hormone, pre proparathyroid hormone, - 31 through - 7, contains a leader or signal sequence. Despite differences in sequence of the parathyroid hormone signal region and other precursor-specific sequences, this region of the molecule possesses biological properties related to intracellular transport and metabolism that appear to be universal for precursor forms of many, if not all, peptide hormones and other secreted proteins. In contrast, the amino-terminal portion of the secreted form of the molecule, sequence region 1-34, has an amino acid sequence that is homologous to that of several peptide hormones, including ACTH, alpha MSH, beta-MSH, and beta-lipotropin. Yet the biological "message" conveyed by this peptide sequence appears unique to parathyroid hormone. Directions have now been established for the design of hormone inhibitors and for analogs of enhanced biological activity and perhaps even analogs possessing an altered spectrum of biological properties. The rapid advances that are occurring in techniques for peptide synthesis, purification, and analysis; in the variety, sensitivity, and specificity of the increasing number of bioassays; and in the elucidation of peptide and protein conformation may provide further important new directions for analog design. Extension of these investigations of structure and function over the next several years should yield a more sophisticated understanding of the mode of hormone action. In such studies lies the promise of generating highly refined and perhaps clinically useful analogs of parathyroid hormone. PMID- 6276848 TI - Clinical implications of preinvasive and small invasive breast carcinomas. PMID- 6276849 TI - Growth and cell kinetic measurements in human tumors. PMID- 6276850 TI - The concept and implications of multicentricity in breast carcinoma. PMID- 6276852 TI - Uterine mesenchymal tumors. PMID- 6276851 TI - Diagnostic electron microscopy. II. Fabry's disease: use of biopsies from uninvolved skin. Acute and chronic changes involving the microvasculature and small unmyelinated nerves. PMID- 6276853 TI - Failure to thrive. AB - Reviewing the chronological evolution of our understanding of why some children fail to thrive without obvious organic cause permits us to develop a rational contemporary approach to diagnosis and management. Originally recognized as a phenomenon of children living in institutions, it later became clear that it could occur in children living with their families. While emotional deprivation was at first emphasized as the principal determinant of growth failure, it is now apparent that the major mediating mechanism is insufficient caloric intake. Thus prior to nutritional recovery it may be difficult to segregate whether emotional symptoms in the child reflect the causes or effects of malnutrition. With rare exceptions, an etiologic diagnosis can be suspected on the basis of history, physical examination, and family assessment. Nonspecific investigative screening of such children has proved to be without value. Successful management depends on recognition of the supernormal caloric intake required to induce rapid catch-up growth. Aggressive hyperalimentation combined with intensive emotional stimulation and often with active family participation offer the best chances of a successful outcome. PMID- 6276854 TI - Plasma opioids in the first hours of life. PMID- 6276856 TI - The mental Health Association in a changing world. PMID- 6276855 TI - Association of coronavirus infection with neonatal necrotizing enterocolitis. AB - From the clustered occurrence of numerous cases of necrotizing enterocolitis in newborns, it was possible to associate this disease significantly with infection due to coronavirus-like agents. Prematurity or low birth weight did not seem to affect the development of the disease, at least during the present epidemic. However, associated gas-producing bacteria could influence its severity and play a role in the appearance of pneumatosis. In many aspects the human disease is reminiscent of experimental necrotizing enterocolitis obtained by infection of germ-free newborn animals, as reported in the literature. PMID- 6276857 TI - The susceptibility of enteropathogenic and non-enteropathogenic porcine Escherichia coli strains to polymyxins and other antibiotics. PMID- 6276858 TI - [Acute myeloblastic leukemia and hepatoma coexisting in the same patient]. PMID- 6276859 TI - [Chronic neurological syndrome resulting from intoxication with metallic uranium (author's transl)]. AB - In 1972, a male patient, then 44-year-old, developed foot cramps and leg pain together with increasing gait disorders and a tendency to fall backwards. In 1976, an extrapyramidal syndrome with ataxia, nystagmus and peripheral neuropathy was discovered on neurological examination. The symptoms persisted and remained of obscure origin until 1979, when a detailed aetiological enquiry disclosed that during at least the first three years of his illness the patient had had on his desk a bar of metallic uranium which he frequently handled. Stool analysis showed significant uranium levels. In the absence of any other cause, it would appear that the neurological syndrome was due to intoxication with uranium. Although this had not been previously described in man, there are occasional reports of uranium toxicity in laboratory animals. PMID- 6276860 TI - [Reactivation of Epstein-Barr virus during primary cytomegalovirus infection in the healthy adult]. PMID- 6276861 TI - [Human leukemic-cell protein-kinases. 1 - Non-granulocytic leukemias (author's transl)]. AB - The study of human leukemic-cell protein-kinases is justified by two types of arguments: on one hand, abnormal protein-kinases have been found in various malignancies, and on the other the products of virus transforming genes have repeatedly been identified as protein-kinases. Using cytosolic and particulate extracts of normal human lymphocytes we measured protein-kinase activities, then following partial purification by DEAE and phosphocellulose chromatography the isoenzymes of cyclic AMP dependent and independent protein-kinases and casein kinases were determined in order to establish a profile (12 normal subjects). The same methodology was applied to the lymphocytes of 5 patients with chronic lymphocytic leukemia (CLL) and 3 patients with acute lymphoblastic leukemia (ALL). Compared to normal lymphocytes, the specific activity of cytosolic and particulate extracts from the leukemics was higher for histone and casein kinases, following elimination of an inhibitory activity present in the crude extracts. Studies of the isoenzymes showed, in some individuals, the presence in both cytosolic and particulate extracts of two important cyclic AMP dependent histone-kinase activities, which were very low or absent in normal lymphocyte extracts. In the particulate extracts we found a constant increase in casein kinase activities concerning essentially one of the two isoenzymes present. Also, the ratio of the different isoenzymes separated by chromatography was considerably modified with regard to both histone and casein-kinases. These quantitative and qualitative abnormalities were present in some CLL cells and in non-granulocytic acute leukemic cells. They resembled the modification reported during normal lymphocyte stimulation by phytohemagglutinin, and also seemed to reflect the intensity of cellular replication and to some degree the progression of the disease. PMID- 6276862 TI - Scintigraphic myocardial imaging with 99mTC-labelled pyrophosphate of experimentally produced cardiomyopathy in dogs. AB - Scintigraphic examination of the myocardium using 99mTc-labelled pyrophosphate was carried out in 10 dogs with experimentally produced cardiomyopathy. This was brought about by intravenous administration of high doses of adrenalin and theophylline. The scan was positive in 8 out of 10 dogs. "Hot" foci were very extensive. The degree of accumulation was however low (2+). Histological examination of the myocardium using the light microscope showed only scarcely distinguishable damage to the tissue without the presence of necrosis. ECG examinations were normal in all cases. By means of 99mTc-labelled pyrophosphate even very small myocardial disorders can thus be detected. This fact may be of clinical importance for an early diagnosis of heart lesions. PMID- 6276863 TI - Pyrophosphate scan of the temporarily ischemized dog myocardium. AB - In 9 dogs a transient myocardial ischemia was provoked using complete occlusion of the ramus interventricularis anterior of the left coronary artery. The occlusion was removed after 5, 10 or 15 min. Four hrs after removal of the occlusion a scan of the myocardium was carried out using 99mTc-labelled pyrophosphate. In 7 out of 9 dogs under study the scan was markedly positive, in 2 dogs negative. ECG demonstrated ischemic changes practically in all dogs; the changes became normal after removal of the occlusion, namely in 5 to 35 min. The histological examination of the tissue demonstrated in all 9 dogs only a slight impairment of the myocardium. PMID- 6276864 TI - Cytomegalovirus: mastering a problem virus. PMID- 6276865 TI - [Effect of polymyxin B inhalation test on blood histamine levels in bronchial asthma]. PMID- 6276866 TI - Influence of sulfated and unsulfated cholecystokinin octapeptide on conditioned feeding behaviour in rats. PMID- 6276867 TI - Effect of beta-endorphin on the thermal excitability of preoptic neurons in the unanesthetized rabbit. AB - The opioid peptide, beta-endorphin (beta-E), will promote changes in body temperature when injected into the brain. It is possible that beta-E alters body temperature by affecting the activity of thermoregulatory neurons in the preoptic anterior hypothalamus (POAH). Single unit activity in the POAH was recorded in unanesthetized rabbits while radiant heat was applied to the dorsal skin. Beta-E was then microinjected into the POAH, and the peripheral heating was repeated. Seventy-seven percent of the POAH neurons were responsive to skin heating. Beta-E and equal excitatory and inhibitory effects on warm-excited and warm-inhibited neurons. Four of six warm-excited neurons were converted to warm-inhibited or unresponsive following beta-E injection. Six out of ten warm-inhibited neurons were converted to warm-excited or unresponsive by beta-E. Beta-E-induced shifts in thermal excitability of POAH neurons may be responsible for the ability of POAH injections of beta-E to elevate body temperature in the rabbit. PMID- 6276869 TI - Central and peripheral injections of ACTH (1-24) reduce fever in adrenalectomized rabbits. AB - Central administration of ACTH (1-24) reduces fever in normal rabbits in doses that have no effect on afebrile body temperature. Previous experimental and clinical reports indicate that peripheral administration of both ACTH and corticosteroids reduces fever, and since central injection of corticosteroids can also lower fever it might be that the antipyretic effect of intracerebroventricular (ICV) ACTH (1-24) is due to adrenal stimulation. To learn whether this endogenous central peptide can produce antipyresis independently, ACTH (1-24) was injected ICV in bilaterally adrenalectomized (ADX) rabbits made febrile by IV injections of leukocytic pyrogen (LP). ACTH (250 ng) given ICV reduced fever in these animals and had a slight hypothermic effect when given to the same rabbits when they were afebrile. Doses of 25-75 ng reduced fever without influencing normal body temperature. Intravenous injections of ACTH (2.5 micrograms) also lowered fever caused by IV LP in ADX rabbits. The present findings raise the possibility that release of endogenous central ACTH, and perhaps entry into the brain of circulating ACTH, the release of which is known to increase in fever, limits the magnitude of the febrile response by influencing central temperature controls. PMID- 6276868 TI - Abdominal vagotomy does not block the satiety effect of bombesin in the rat. AB - Bombesin (2-16 microgram-kg-1, intraperitoneally) inhibited food intake in rats after abdominal vagotomy. Since the same vagotomized rats did not respond to the octapeptide of cholecystokinin (1-8 micrograms-kg-1, intraperitoneally), these data are decisive evidence (1) that bombesin does not produce satiety by releasing endogenous cholecystokinin and (2) that vagal afferents are not necessary for the satiety effect of bombesin. PMID- 6276870 TI - In vivo and in vitro release of ACTH by synthetic CRF. AB - The 41-residue corticotropin releasing factor (CRF) was synthesized by the solid phase method. The synthetic CRF and arginine vasopressin (AVP) were examined for ACTH releasing activity and effects on the release of 5 other pituitary hormones in vivo and in vitro. Injection of the CRF into pharmacologically blocked rats increased plasma corticosterone levels in a dose-related manner. The minimum effective dose was 1.6 x 10(-12) mol/100 g body weight. CRF also significantly stimulated release of ACTH-like immunoreactivity in a dose-related manner from rat pituitary quarters beginning at a concentration of 10(-9) M. AVP, a peptide known to have CRF activity, exhibited slightly lower corticotropin releasing activity than the CRF at equimolar dose levels. Secretion of other pituitary hormones was not appreciably altered by either the CRF or AVP. PMID- 6276871 TI - Cholecystokinin and bombesin act independently to decrease food intake in the rat. AB - The satiating effects of cholecystokinin-octapeptide (CCK-8) and bombesin (BBS) when injected alone and in combination were compared in intact rats. When injected alone, both CCK-8 and BBS elicited a dose-related decrease of 30-minute food intake. Injections of BBS were less potent than the equivalent doses of CCK 8 in producing satiety. BBS reached an asymptotic level of suppression of approximately 40 percent at a dose of 2 micrograms/kg, whereas injections of 4 micrograms/kg of CCK-8 resulted in a 72 percent suppression of food intake. When the two peptides were administered in a single injection, the resulting suppression of food intake was equivalent to that which would be predicted if their effects were completely additive. These results support the hypothesis that CCK-8 and BBS act via independent mechanisms to induce satiety. A preliminary model of peptidergic satiety, based on this hypothesis, is proposed. PMID- 6276872 TI - The specificity of angiotensin II receptor binding in rat brain. AB - 125I-angiotensin II (125I-AII) binding was examined in the hypothalamic-thalamic septal-midbrain (HTSM) region of HLA-Wistar rats in the presence of CNS-active agents. Angiotensin I, II, and III and saralasin competed for 125 I-AII binding, whereas structurally unrelated peptides such as arginine and lysine vasopressin, oxytocin, LHRH, TRH, bradykinin, and substance P did not. In contrast, ACTH and neurotensin exhibited a weak, dose-dependent competition for 125 I-AII binding. The relative potencies of AII, AI, neurotensin and ACTH were 100:1:0.1:0.05, respectively. Neurotensin and ACTH competition was not additive with AII suggesting interaction at shared binding sites. Most importantly, a wide variety of other CNS active agents such as methyldopa, naloxone, catecholamines, clondidine, and reserpine, failed to inhibit 125 I-AII binding, thus further defining the specificity of the CNS AII receptor. PMID- 6276873 TI - Thalamic mediation of impulses arriving from the substantia nigra. AB - Unilateral injection of carbachol (CCh), 3 mu, to the pars reticularis (SNr) of the substantia nigra evoked electroencephalographic epileptiform discharges in the ventro-lateral thalamic nucleus (VLTh), nucleus caudatus (NC( and substantia nigra (SN) in rabbits. Administration of atropine, 2.5 micrograms, to the VLTh, prior to the CCh injections, completely blocked the epileptiform discharges in all the examined structure. Injections of picrotoxin, 1 microgram, on the border line between the SNr and pedunculus cerebri (PC) evoked also epileptiform discharges, which were blocked by an intrathalamic injection of GABA, 40 micrograms. It is concluded that both acetylcholine and GABA receptors are present both in the SN and VLTh, that between SN and VLTh there exists functional connection represented most probably by the nigro-thalamic pathway, and that GABA receptors of the VLTh are linked with the endings of the nigro-thalamic pathway while the cholinergic thalamic receptors are connected with another thalamic input. PMID- 6276874 TI - Spontaneous urinoma with peripheral neuropathy. PMID- 6276875 TI - Clonidine for opiate withdrawal in the chronic pain patient. PMID- 6276876 TI - Host age and major histocompatibility genotype influence on Rous sarcoma regression in chickens. AB - The influence of host age and B (major histocompatibility complex) genotype on the fate of Rous sarcoma virus (RSV)-induced tumors was investigated. White Leghorn lines 61 and 151, developed at the Regional Poultry Research Laboratory (RPRL) at East Lansing, MI, and their crosses (F3 to F5) segregating for the B genotype were used. Preliminary experiments with noninbred White Leghorns showed that chickens RSV-inoculated at 4 weeks of age had a significantly lower incidence of tumor regression than chickens RSV-inoculated at 6, 8, 10, 12, and 14 weeks of age. The (61 X 151) F5B2/B2 and B5/B5 chickens inoculated at 6 weeks of age had a lower incidence of tumor regression than (61 X 151)F3 and F4, B2/B2 and B5/B5 chickens RSV-inoculated between 58 and 88 weeks of age, respectively. Some (61 X 151) B2/B2 chickens inoculated between 58 and 88 weeks of age regressed their tumors while all chickens of corresponding genotype inoculated at 6 weeks of age died with tumors. the (61 X 151) B2/B2 chickens had a higher incidence of tumor regression than B5/B5 chickens regardless of age. The genetic RSV-inoculation on the fate of RSV-induced tumors. PMID- 6276877 TI - Rehabilitation of the leg amputee in the community. PMID- 6276878 TI - Audit of a neurological follow-up clinic. PMID- 6276879 TI - Scheduling Pap smears for asymptomatic women. PMID- 6276880 TI - [Functional role of the superoxide radical and superoxide dismutase in formed blood elements]. PMID- 6276881 TI - Renal transplantation in children. PMID- 6276882 TI - Mechanism of allopurinol action in calcium oxalate stone formers. PMID- 6276883 TI - Predictive value of 99mTc pyrophosphate bone scintigraphy for vitamin D trials in uraemia. AB - The usefulness of 99mTc-Pyrophosphate (99mTc-PPi) bone scintigraphy was evaluated in the follow up of 21 haemodialysed patients without clinical or radiological evidence of osteodystrophy. 99mTc-PPi bone scintigraphy was semi-quantitatively analysed using Fogelman's score. Patients were randomised to receive vitamin D analogues (1 alpha hydroxyvitamin D3 or dihydrotachysterol, n = 12) or to serve as controls (n = 9), both groups being given oral calcium supplements. Bone scintigraphy deteriorated in patients only on calcium therapy but not in patients treated by vitamin D-analogues. Vitamin D therapy reduced secondary hyperparathyroidism in all cases but induced rapid intoxication with normal doses in 4 of the 12 treated patients. Since intoxicated patients had significantly lower Fogelman's score than the patients who tolerated the treatment well, 99mTc PPi bone scintigraphy is proposed as a screening test before vitamin D-analogues trials. PMID- 6276884 TI - T/t locus, its role in embryogenesis and its relation to classical histocompatibility systems. PMID- 6276885 TI - Enzymic diversification of the sodium pump in sheep red cells. AB - It is well-established that sheep are genetically dimorphic with respect to the K+ content of their mature red cells and that high-K+ (HK) and low-K+ (LK) red cells differ with respect to the activity and kinetic properties of the sodium pump and Na+-ATPase. In contrast, reticulocytes of both types have a high K+ content and are similar with respect to their pump and Na+-ATPase kinetic characteristics. Reticulocytes differ, however, from mature cells of either HK or LK sheep. The first part of this paper describes a comparative study of the side specificity of Na+ and K+ interactions with Na--ATPase of inside-out membrane vesicles prepared from mature HK and LK red cells. The results indicate that the genetic difference between the two is expressed as differences in apparent affinities for both Na+ and K+ and that these differences are evident at the cytoplasmic as well as at the extracellular membrane surface. The second part of this paper describes a study of the nature of the maturation-associated changes in the sodium pump system of sheep reticulocytes. Maturation was followed both in vitro (long-term incubation at 37 degrees of reticulocytes) and in vivo (changes in density gradient fractionated HK cells followed at intervals during recovery from massive bleeding. The results support the conclusion that the dimorphism in the sodium pump of sheep red cells is a result of specific changes in Na+-ATPase and that these changes proceed differently during maturation of the two types of sheep red cells. PMID- 6276887 TI - Studies of the mechanism of bacteriophage T4 DNA encapsidation. PMID- 6276886 TI - Modulators of red cell Na/K pump rates. PMID- 6276888 TI - Interactions between the maturation protein gp17 and the single-stranded DNA binding protein gp32 initiate DNA packaging and compete with initiation of secondary DNA replication forks in phage T4. AB - Since T4 DNA, which is being packaged is actively replicating and recombining, we have asked whether replication and recombination proteins interact with maturation proteins. We report here on genetic evidence for interaction of the single-stranded DNA binding protein gp32 with the maturation protein gp17. Based on these and other results we suggest a model in which DNA packaging is initiated from recombinational intermediates.U PMID- 6276889 TI - Studies on the function of gene 49 controlled endonuclease of phage T4 (endonuclease VII). PMID- 6276890 TI - Theoretical evidence for the significance of certain types of DNA secondary structure in the genomes of phi X174, G4, fd, SV40 and the plasmid pBR322. AB - The significance of certain types of DNA secondary structure was assessed on a purely theoretical basis by analyzing their distribution over the entire genomes of bacteriophages phi X174, G4, fd, the eucaryotic virus SV40 and the E. coli plasmid pBR322. A computer program was designed to search the translated and non translated (intercistronic) regions of these genomes for the potential of forming perfect hairpins (perfectly base paired stems: no G-T pairing; no interruptions) with small loops. The number of found occurrences was then compared to the statistically expected number. The results reveal a significant excess in each genome of perfect hairpins with stem length greater than 7 and end loops less than or equal to 20 over that expected given their nucleotide composition. For stem length of 5 to 7 the same observation holds true for the intercistronic regions, but for translated regions the number of such structures is below expectation. This suggests an evolutionary selection for these structures. PMID- 6276891 TI - Psychopharmacology of clonidine: an introduction. PMID- 6276893 TI - Clonidine: antidepressant potential? PMID- 6276894 TI - Hyperphagia and weight gain in monkeys treated with clonidine. PMID- 6276892 TI - Clonidine and the primate locus coeruleus: evidence suggesting anxiolytic and anti-withdrawal effects. AB - One may conclude from this series of studies and the data of others reviewed at this symposium that NE systems are involved in fear or anxiety and in mediating the actions of opioids and endorphins. It is rational pharmacotherapy, therefore, to utilize anatomically and neurophysiologically characterized receptors to produce those desired specific effects on NE systems that are expected to reduce anxiety. These same receptor interactions could also be used to antagonize side effects or to reverse withdrawal syndromes via alternative receptor sites. This strategy of using different receptors to produce synergistic or antagonistic effects on a common neuronal pathway has benefited the treatment of pathological anxiety, pain, drug withdrawal symptoms, and other classical psychosomatic conditions. It should be emphasized, of course, that the LC-noradrenergic system has a much broader function than the mediation of fear or anxiety, and that the endorphins, opiates, and clonidine influence many other systems, as do their abstinence syndromes. Anatomical projections of the LC and similar central NE systems also produce the observed effects. Detailed studies are needed to discover precisely how the abnormalities observed in numerous biochemical systems [Eidelberg, 1976] are related to the alterations in NE function we have studied and discussed in this chapter. Finally, careful clinical studies are needed to determine, define, and delimit the potential therapeutic uses of clonidine in the areas suggested by the preliminary studies reviewed here. PMID- 6276895 TI - Antinociceptive actions of clonidine. PMID- 6276896 TI - Antinociceptive mechanism and acute and chronic behavioral effects of clonidine. PMID- 6276897 TI - Neuroanatomical sites of action of clonidine in opiate withdrawal: the locus coeruleus connection. PMID- 6276898 TI - Brain sites for the antihypertensive action of clonidine. AB - In conclusion, I reviewed recent literature that directly investigated the site and mechanism of action of clonidine in the central nervous system. One can interpret these data in the following manner. In the medulla, clonidine is acting as an alpha-adrenergic receptor antagonist that inhibits excitatory input to the sympathetic nervous system. In the anterior hypothalamus, clonidine is acting as an alpha-adrenergic receptor agonist that excites an inhibitory pathway which inhibits excitatory input to the sympathetic nervous system. Both actions produce the same final response; that is, diminished sympathetic outflow from the central nervous system. PMID- 6276900 TI - Anatomical mapping of clonidine (alpha-2 noradrenergic) receptors in rat brain: relationship to function. PMID- 6276899 TI - Clinical utility of clonidine in opiate withdrawal: a study of 100 patients. PMID- 6276902 TI - 3H-clonidine and 3H-p-aminoclonidine interactions in vitro with central and peripheral alpha 2-adrenergic receptors. PMID- 6276901 TI - The neuroanatomy and pharmacology of the nucleus locus coeruleus. AB - Several points are particularly relevant to an understanding of the effects of clonidine in the brain: Clonidine appears to act preferentially at alpha-2 adrenergic receptors and, in systemic doses of less than 50-100 microgram/kg, to exert its effects at autoreceptors, which initially decrease noradrenergic neuronal firing rates, NE release, and NE turnover. This action results in functional decreases of the usual effects of these neurons at their post synaptic alpha-1 and beta-adrenergic receptor-mediated projections, which are widely distributed throughout the limbic system, cerebrum, cerebellum, and spinal cord. Higher doses of clonidine appear to produce agonist effects at alpha-1 adrenergic receptors, counteracting the effects of decreases in firing rates and turnover. However, decreased function at beta receptors would continue after these higher doses owing to continued inhibition of neuronal activity and the lack of direct effects of clonidine on beta receptors. Receptors for GABA, endorphins, substance P, and ACh on LC neurons provide an anatomical and physiological basis for interactions among systems utilizing these substances as neurotransmitters in that they act on a common final noradrenergic pathway. These noradrenergic pathways, in turn, also project to and mediate functional changes in areas utilizing other neurotransmitters. Although these changes affect numerous types of behavior and responses that are measurable by psychopharmacologists, it is the neurons of the locus coeruleus and other noradrenergic neurons with similar receptor combination that provide the entry point to this NE system. Studies of the LC, therefore, may help to determine the nature of these specific effects and to characterize the effects of clonidine and other alpha-2 agonists on the brain. PMID- 6276903 TI - Mechanisms of antihypertensive action of clonidine in relation to its psychotropic effects. PMID- 6276904 TI - Psychotropic actions of clonidine. PMID- 6276905 TI - Desglycyl-8-arginine vasopressin affects regional mouse brain cyclic AMP content. PMID- 6276906 TI - DNA and protein synthesis in normal and transformed lymphocytes exposed to abrin. AB - The dose dependent effects of abrin, a toxic D-galactose binding plant lectin, on 3H-TdR and 14C-leucine uptake are studied in normal and Epstein Barr Virus (EBV) transformed lymphocyte cultures. Results show that while abrin is highly toxic to both the DNA and protein synthesis in EBV lymphocytes, some toxicity to the normal cells is also seen. It is postulated that lymphocyte DNA synthesis is affected by ribosomal shutdown induced by the abrin. PMID- 6276907 TI - Neuromelanin: a hypothetical component of bioelectronic mechanisms in brain function. AB - Several lines of evidence suggest the involvement of bioelectronic mechanisms in brain function. Although no components of such mechanisms have been identified, neuromelanin is a likely candidate. The functional significance of neuromelanin is supported by considerable published work, herein reviewed. Certain electronic properties of melanin, notably in stable free radical characteristics and its semiconduction, may be influenced by alpha-MSH, ACTH 4-10, and melatonin. Neuron function may be influenced in turn by the effect of semiconduction on synaptic transmission. Thus neuromelanin may be an important part of a complex neural modulation system. PMID- 6276908 TI - Ascorbic acid as a thiolprive: ability to induce immunity against some cancers in mice. AB - Immunological studies are reported showing that ascorbic acid, like selected sulfhydryl inhibitors, can induce immunity against some cancers in mice. Accompanying this immunizing action are changes in the surface structure of the cancer cells, as revealed by scanning electron microscopy. Electron spin resonance measurements show that the ascorbate anion free radical is readily produced in oxygenated cancer tissue and that this radical can react with sulfhydryl groups which are free radical scavengers. It is proposed that ascorbate acts as an effective thiolprive in oxygenated cancer tissues. This action is thought to lead to the observed changes in the cancer cell surface structure and to the concomitant immunological response. PMID- 6276909 TI - [Changes in cerebrospinal fluid and plasma cyclic nucleotides following amygdaloid kindling and kindled convulsions in cats (author's transl)]. PMID- 6276910 TI - From institutional to community care. PMID- 6276911 TI - Stress-induced plasma beta-endorphin immunoreactivity may predict postoperative morphine usage. AB - The relationship between stress and human behavior is studied using general surgery as a stress paradigm. As indices of arousal, presurgery and surgery plasma beta-endorphin and cortisol immunoreactivity are assessed. Behavioral analysis is restricted to the measurement of total morphine usage during the first 24 postoperative hours under standard PRN (as needed) clinical orders. Individual patient morphine requirements vary widely (12-56 mg). Both presurgery and mean surgery plasma beta-endorphin levels significantly predict morphine requirement, and similar, although not so strong, correlations are found for cortisol. Patient age is also found to be negatively correlated with morphine requirement. When multiple regression analysis is used, the variables of plasma beta-endorphin and age predict 70% of the variance in individual morphine requirements. PMID- 6276912 TI - Structure and functions of the Heidelberg University organisation for after-care of cancer patients. AB - The functions of the psycho-social after-care centre are: care of patients co operation with self-help groups, and training of therapists in psycho-social work. We describe the psycho-social therapist's work with out-patients and in patients (e.g., in surgical wards; children's hospital), including the various focuses of attention in conversations with patients. Frequently the family has to be involved in helping to cope with the illness. Some account is given of our work with self-help groups, and of the training courses developed for therapists working with cancer patients. Finally, we discuss the question of individual suitability for work with cancer patients and for related research projects. PMID- 6276914 TI - [Hemoglobinopathies]. PMID- 6276913 TI - Endorphinergic systems and the response to stress. AB - Endogenous opioid peptides (EOPs) are extensively disturbed throughout the CNS and found in the CSF, pituitary, plasma and many peripheral organs of animals and man. Their intracerebroventricular administration produces a spectrum of morphinomimetic effects. Biochemical evidence for an acute stress-evoked release of brain, spinal cord and pituitary pools of EOPs has accumulated and the sensitivity of both adult and developing endorphinergic systems to chronic stress has been shown. EOPs are involved in the mediation of the elevations in nociceptive threshold, core temperature, exploration and grooming behaviour elicited by stress. A role in the attenuation of anxiety is also indicated. Finally an endorphinergic control of pituitary secretion and the activity of peripheral organs under stress has been demonstrated. EOPs are apparently of general importance in the response to stress in both animals and man. PMID- 6276915 TI - Viral hepatitis. PMID- 6276916 TI - [The retrograde jugulography for the diagnosis of glomus tumours in the skull base (author's transl)]. PMID- 6276917 TI - [Simple application of tissue-adhesive contrast-medium mixtures for transversal tumour embolization (author's transl)]. PMID- 6276918 TI - [Experimental nuclear medical examinations with the replacement of extended defects on the long bones (author's transl)]. PMID- 6276919 TI - [Proton-translocating ATPase of Escherichia coli (author's transl)]. PMID- 6276920 TI - [Application of plant virus studies in genetic engineering]. PMID- 6276921 TI - [Na+-Ca2+ exchange system and its physiological role (author's transl)]. PMID- 6276922 TI - [Mechanisms of action of and resistance to colistinpolymyxin group antibiotics including resistance of colitin-producing bacterium, Bacillus colistinus to colistin (author's transl)]. PMID- 6276923 TI - [Biogenesis of viral glycoproteins and role of sugar chains in biological functions of virus (author's transl)]. PMID- 6276924 TI - [Oncogenesis by RNA tumor virus (author's transl)]. PMID- 6276925 TI - Platelet-vessel wall interactions: effects of platelets and plasma on the antiaggregatory activity and 6 keto-PGF1 alpha production in isolated perfused aortas. AB - An experimental model for the study of platelet-vessel wall interactions has been developed, based on perfusion of rat platelet rich plasma (PRP) through isolated rat aortas. In the perfused PRP, platelet aggregation was inhibited and levels of 6 Keto PGF1 alpha and cAMP were elevated over the values found in non perfused PRP. When PPP or buffer were perfused through the isolated artery, elevations of 6 Keto PGF1 alpha levels in the perfusate were smaller (in perfused PPP) or of shorter duration (in both perfused PPP and buffer). The presence of platelets in the perfusion fluid thus enhanced the formation of Prostacyclin by the arterial wall. Levels of 6 Keto PGF1 alpha in PRP obtained from aspirin-treated animals and in PRP from normal animals, both perfused through normal aortas, were the same, and also levels of the above metabolite in normal PRP perfused through aortas of aspirin-treated animals did not differ from those found in non perfused PRP. It is concluded, from these data, that PRP does not stimulate PGI2 formation in perfused aortas by providing cyclic endoperoxides. The experimental model developed allows the study of interactions between normal platelets and aortas from experimentally treated animals or viceversa. PMID- 6276927 TI - [Neutrophil NADH oxidase and myeloperoxidase activity in psoriasis]. PMID- 6276928 TI - [Vaccination against poliomyelitis. Formation of immunity evaluated according to the results of serological surveys]. PMID- 6276926 TI - Histamine, endogenous prostaglandins and cyclic nucleotides in the regulation of airway muscle responses in the guinea pig. AB - Indomethacin (30 mg/kg, i.p.) reduced pulmonary resistance in guinea pigs but did not affect their sensitivity to histamine. This treatment preferentially reduced the generation of PGE2 by isolated tracheal preparations. The ratios of PGF2 alpha/PGE2 before and after treatment were 1/1 and 6/1, respectively. Chronic indomethacin treatment (30 mg/kg, i.p., twice a day for 4 days) increased histamine sensitivity in vivo 2 fold while a longer treatment (10 days) was without effect. The efficacy of histamine and the potency of isoproterenol in tracheal tissues were unaffected by either treatment. Indomethacin (17 microM for 30 min) relaxed tracheal tissues but not bronchial tissues. Responses of both tissues to contractile agonists were potentiated after indomethacin treatment. The efficacy of histamine was smaller in bronchi than in tracheas. Similarly, PGE2, PGI2 and isoproterenol were less potent in bronchi. Basal amounts of cyclic AMP were higher in bronchi than in tracheas; indomethacin did not affect the basal amounts of cyclic AMP in tracheal tissues but reduced them in bronchial preparations. Histamine elevated cyclic AMP content in both preparations; this elevation was reduced by indomethacin. While prostaglandins play a role in modulating airway responses in vitro, their role in airways in normal animals in vivo is more difficult to demonstrate. PMID- 6276929 TI - [The insulinoma case diagnosed by the use of glucagon test (author's transl)]. PMID- 6276930 TI - [Standard silica dust used in studies of fibrogenetic effect of industrial dusts (author's transl)]. PMID- 6276931 TI - [The morphologic and ultrastructural evaluation of biological effect of silicon dust proposed as a standard dust (author's transl)]. PMID- 6276932 TI - [Assessment of RIA kit for measuring pregnancy-specific beta 1 glycoprotein (SP1) and its clinical significance as a tumor marker (author's transl)]. PMID- 6276933 TI - The role of GABA in analgesia and drug dependence. PMID- 6276934 TI - [Benign hepatic tumors]. AB - A total of 30 cases are presented, of benign hepatic tumours. As a result of the confrontation of data from the literature with the personal experience of the authors has evidenced the diagnostic difficulties in patients with benign hepatic tumors, especially because these can be erroneously considered as either malignant tumours or as liver cirrhosis. The clinical and the para-clinical pictures are not suggestive in all the cases, and the necessity arises of histopathologic confirmation which can be achieved by puncture and cytologic examination, or by intra-surgery biopsy. This is particularly useful because some of the benign hepatic tumours may benefit from a radical surgical treatment. PMID- 6276935 TI - [Phyllode tumors: anatomoclinical and therapeutic aspects (5 clinical cases)]. AB - The phyllodes tumour is a bi-tissular mammary tumour. It is essentially benign but potentially sarcomatous, presenting multiple clinical and histologic-aspects, and raises difficult problems of diagnosis and therapeutic attitude. Four histological types are described, from the adenofibromatous type to the sarcomatous one. The notion of local malignancy is discussed, related to the marked recidivating character of the tumour. The morphologic substrate of the recidivating character is the result of the absence of a true capsule of the tumour. The treatment is differentiated according to the clinic, histologic, and recidivating aspects, and consists in either sectorectomy or breast amputation. Five cases are presented, of phyllodes tumour, each of them with particular characteristics of evolution, and imposing differentiated therapeutic attitudes. PMID- 6276936 TI - [Controversies in nutrition]. PMID- 6276937 TI - [Role of A virus in the genesis of chronic hepatitis and distribution of its antibodies in the current adult Spanish population]. PMID- 6276938 TI - [Calcitonin: a hormone with multisystemic effects]. PMID- 6276939 TI - [Conceptual aspects of myasthenia gravis]. PMID- 6276940 TI - [Pulmonary carcinoma with inappropriate ADH and ACTH secretion]. PMID- 6276941 TI - [Use of sodium Tc-99m pertechnetate in the diagnosis of Meckel's diverticulum. Apropos of a case]. PMID- 6276942 TI - [Sensory-motor neuropathy associated with a benign monoclonal gammapathy. Apropos of 2 observations]. PMID- 6276943 TI - [Current therapeutic aspects of myasthenia gravis]. PMID- 6276944 TI - [Intra- and extracellular cyclic AMP in VERO cells in response to the thermolabile enterotoxin of Escherichia coli]. PMID- 6276945 TI - [Morphinics receptors and endorphines (author's transl)]. PMID- 6276946 TI - [Diagnostic aspects of insulinoma. Apropos of 2 cases]. PMID- 6276947 TI - Metastatic CNS disease in small cell carcinoma of the bronchus: the role of radiotherapy and chemotherapy. PMID- 6276948 TI - Malnutrition in lung cancer: incidence, prognostic implications, and pathogenesis. AB - Malnutrition and weight loss are common in patients with lung cancer. Weight loss is an independent prognostic factor for survival in lung cancer treatment studies. Metabolic disturbances probably play a dominant role in weight loss in these patients rather than reduced food intake. The identification of the pertinent etiologic metabolic abnormalities and development of specific therapeutic intervention should be goals for future research. PMID- 6276950 TI - Angiotensin converting enzyme activity in human serum: relationship to enzyme inhibitor in vivo and in vitro. AB - Quantitative relationships between angiotensin converting enzyme activity in human serum and concentration of an enzyme inhibitor, the active diacid metabolite of MK-421 [N-(S-1-carbethoxy-3-phenylpropyl)-S-alanyl-S-proline maleate], were defined. Dose-response curves were generated in vitro by adding the diacid metabolite (DM) to serum (ID50 = to 6x10-9 M); l0-15% of initial activity was nonsuppressible. DM an converting enzyme activity were also measured in serum from hypertensive subjects receiving oral MK-421. The dose-response curve for serum samples following single doses of MK-421 (ID50 = 6x10-9M), was similar to curves generated in vitro while the curve for sample following chronic therapy suggested a small decrease in inhibitory potency. PMID- 6276949 TI - Follow-up of a randomized trial for oat cell carcinoma evaluating the efficacy of peripheral intravenous nutrition (PIVN) as adjunct treatment. AB - A randomized trial was initiated to compare the effects of peripheral i.v. nutrition (PIVN)-associated chemotherapy (adriamycin, vincristine, VP-16-213, and cyclophosphamide) versus a chemotherapy control group in patients with oat cell lung carcinoma. Thirty-nine evaluable patients were randomized. The test group included 19 patients, whereas 20 were followed in the control group. PIVN was scheduled each day the patient underwent chemotherapy. Each patient received 1,550 kcal day which included 10% glucose, 20% lipids, and amino acids which may or may not have been mixed in the same infusion bottle. The results show ten PIVN patients presently in complete remission at the end of three courses of treatment, compared to nine over the same time period in the chemotherapy control group. Thirty-three percent of patients are still alive after 15 months after the beginning of treatment. There was no significant difference in either general health or side effects, frequency or duration of complete remission, or survival time. After 1 year of treatment, 8 of 19 PIVN patients are still in complete remission, compared to 7 of 20 patients in the control groups. PMID- 6276951 TI - Significant increases of cyclic AMP and alkaline phosphatase in bronchoalveolar lavage fluids of sheep exposed to asbestos. AB - Exposure to asbestos for six months caused a significant increase of adenosine 3',5'-cyclic monophosphate (cAMP) levels (2-fold in bronchoalveolar lavage (BAL) fluids of sheep with no significant change in guanosine 3',5'-cyclic monophosphate (cGMP) and alkaline phosphatase (AP) activity. When exposure was prolonged for another 3 month-period, the increase in cAMP levels persisted and was accompanied by a 4-fold stimulation of AP activity, and enzyme associated with the surfactant fraction of BAL fluids. PMID- 6276952 TI - Effects of propylthiouracil on MHV-3 virus mouse hepatitis. AB - A study was undertaken to observe the effect of propylthiouracil (PTU) on MHV-3 virus mouse hepatitis. PTU-treatment adversely influenced the course of the hepatitis. This effect was reflected by a significantly higher mortality rate, in spite of the absence of any significant difference of the plasma transaminase activity increase and of the liver virus content. PMID- 6276953 TI - Biphasic amphetamine effects on skeletal muscle contractions: noncholinergic mechanisms. AB - Noncholinergic mechanisms underlying the biphasic effects of (+)-amphetamine on skeletal muscle contractions were studied in the directly-stimulated, cholinergically blocked, rat phrenic nerve-diaphragm preparation. Low (2.7-8.1 x 10(-4)M) and high (10.8-21.6 x 10(-4)M) levels of (+)-amphetamine produced concentration-dependent enhancement and blockade of muscle contractions, respectively, in tissues pretreated with alpha-bungarotoxin, beta-bungarotoxin, or (+)-tubocurarine. These biphasic effects were not modified by tyramine, phentolamine, or propranolol arguing against a mechanism involving a direct or indirect adrenoceptor interaction. Amphetamine did not modify (Na+-K+)-stimulated ATPase activity. Increased [K+] or decreased [Na+] antagonized (+)-amphetamine enhancement of contractions and potentiated blockade; similar effects were observed after high concentration amantadine or procaine pretreatment. K+-free media potentiated (+)-amphetamine facilitation of contractions as did low concentrations of amantadine or procaine. These results suggest that amphetamine may biphasically modify skeletal muscle contractions by interactions involving the movements of Na+ and K+ independent of (Na+-K+)-stimulated ATPase. PMID- 6276954 TI - Prostacyclin: effects on cyclic AMP in bone cells. AB - Prostacyclin (PGI2) increased cyclic AMP in specific rat bone cell populations obtained by sequential collagenase digestion of calvariae. An osteoclastic population, characterized by high acid phosphatase levels and the ability to resorb bone in vitro, was more sensitive as reflected by responses to lower doses of PGI2 (2.3 x 10(-8)M) as well as greater responses to higher doses (3.5 x 10( 4)) of PGI2 than the other populations isolated. The osteoblastic populations, characterized by high alkaline phosphatase levels and the inability to resorb devitalized bone did not respond significantly to PGI2 at doses less than 10(-4)M and increases in cAMP were smaller. Then results suggest a differential effect of PGI2 on osteoclasts and osteoblasts which may be involved in the mode of action of endogenously produced PGI2 in bone. PMID- 6276955 TI - An investigation of the effects of aminoglycoside antibiotics on Na+-K+ ATPase as a possible mechanism of toxicity. AB - An investigation was made of the effect of aminoglycoside antibiotics on Na+-K+ ATPase to determine whether their toxicity might be attributable to an inhibition of this enzyme. Three preparations were used: human erythrocyte ghosts; microsomal fractions of the cortex and outer medulla of the guinea-pig kidney; rat isolated renal tubules. In all three preparations marked inhibition of Na+-K+ ATPase (50-100%) was seen only at very high concentrations of drug (10(-2)M). Although aminoglycoside antibiotics are known to accumulate to high concentrations in vivo, particularly in the renal cortex, it is argued that these findings suggest that an effect on Na+-K+ ATPase is unlikely to be their primary mechanism of toxicity. PMID- 6276956 TI - [Prospective evaluation of diagnostic procedures in patients suspected of malignant hepatomegaly (author's transl)]. PMID- 6276957 TI - [Structure, identification of the genes and integration of the hepatitis B viral genome (author's transl)]. PMID- 6276958 TI - [Psychological components of stress. Part II (author's transl)]. PMID- 6276959 TI - [Herpetic disease in newborn infants. Pathogenetic and preventive aspects]. PMID- 6276960 TI - [Incidence of antibodies against hepatitis virus A (anti-HA) in a non-selected urban population]. AB - Anti-HA antibodies were determined by radioimmunoassay according to the HAVAB ABBOTT technique in 244 subjects, of whom 194 without hepatic involvement, 25 with non B hva and 25 former non B hva patients. Anti-HA anti-bodies were found in 67% of the healthy subjects and 100% of the patients and former hva cases. The incidence of anti-HA antibodies increases with age being in 30% of children and 83.5% of adults. A peak incidence was found between the ages 21 and 30 years. In the district investigated subclinical infection with HVA was frequent, beginning in childhood and up to the age of 30 years. The qualitative HAVAB test is adequate for epidemiological investigations; a quantitative variant is necessary for the diagnosis. PMID- 6276961 TI - [Systemic cellular mediated immunity in children hospitalized with clinical, virological and serological diagnosis of Coxsackie B virosis]. PMID- 6276962 TI - [The problem of multiple breast cancers (author's transl)]. AB - This retrospective study is based on the appearance of bilateral breast cancers - occurring simultaneously or succeedingly -- in patients our therapeutical radiation section. The examination of 368 patients in the years 1978-1981 gave result of 12 cases (3,2%) showing simultaneous bilateral breast cancer and of 14 cases (3,8%) showing succeeding contralateral breast cancer. Therefore multiple breast cancers are not to be regarded as a limited process but as manifestation in a cancerogenic effective range. This is of prime importance to the diagnosis and therapy of breast cancer. PMID- 6276963 TI - Effects of ototoxic diuretics on cochlear Na+/K+-ATPase and adenylate cyclase. PMID- 6276964 TI - Control mechanisms in active chloride transport. PMID- 6276965 TI - Influence of high ceiling diuretics on ion fluxes and cell volume of ehrlich ascites tumour cells. PMID- 6276966 TI - Hyperplastic lesions of the human breast: scanning electron microscopy and a review of current knowledge. AB - Although the etiology of human breast cancer is unknown, certain epidemiological parameters such as risk of breast cancer development. Concurrent, retrospective and prospective studies have shown that one particular component of benign breast disease -- epithelial hyperplasia, may be responsible for the increase in risk. Several classification have been devised for epithelial hyperplasia of the human breast using light microscopy. By scanning electron microscopy epithelial hyperplasia in ducts has several patterns: focal hyperplasia, simple columnar bridging, complex bridging and solid hyperplasia. Although these patterns may be seen in non-cancerous ducts in both malignant and non-malignant cases, alterations in microvillus structure and number have only been seen to date in malignant cases. PMID- 6276967 TI - Use of plaque assays to study thyroglobulin autoantibody synthesis by human peripheral blood lymphocytes. AB - Plaque assays have been used to study thyroglobulin autoantibody synthesis and total immunoglobulin production by cultures of peripheral blood lymphocytes from patients with Hashimoto's thyroiditis. Freshly isolated Hashimoto or normal lymphocytes contained small numbers (7-1300) of total IgG plaque-forming cells (PFC) and total IgM PFC, but specific thyroglobulin antibody PFC was undetectable. After 5-8 days' culture with pokeweek mitogen (PWM), total IgG and IgM PFC were markedly increased to a geometric mean of 10,140 (8414-12,220) IgG PFC and 3450 (163-7534) IgM PFC per 10(6) cultured lymphocytes (95% confidence limits in parentheses). Furthermore, a mean of 63 (22-287) specific IgG thyroglobulin antibody PFC per 10(6) lymphocytes were detectable in cultures of Hashimoto lymphocytes. The IgG thyroglobulin antibody PFC were stimulated by Epstein-Barr virus (EBV) infection, suggesting that EBV infection may be useful in obtaining monoclonal thyroid autoantibodies. PMID- 6276969 TI - Neurophysiological findings among workers exposed to organic solvents. AB - Workers exposed to organic solvents often complain of symptoms indicating central or peripheral nervous system effects. Neurophysiological methods offer a sensitive and noninvasive way to study early impairment in the nervous system. The present communication is a review of neurophysiological studies done at the Institute of Occupational Health, Helsinki. The studies concern workers occupationally exposed to various solvents. Both encephalopathic and neuropathic findings were frequent, especially among patients with solvent poisoning. Certain exposure-response relationships emerged when the prevalence of abnormal electroencephalograms was studied in various groups of workers exposed to different levels of solvents. PMID- 6276968 TI - Neuromuscular transmission in the corpus-fundus of the urinary bladder. An in vitro study in the cat. AB - In muscle strips from the corpus-fundus of the cat bladder the intramural nervous system was activated by field stimulation. The resulting muscle contraction could not be inhibited by atropine, phentolamine or quinidine. These drugs enhanced the response. Tetrodotoxin, methysergide, cyproheptadine and indomethacin blocked the field-stimulation response in a dose dependent manner. The interpretation of these results with references to transmission at the neuromuscular junction of the bladder is discussed. PMID- 6276970 TI - [Pain and pain therapy - neurophysiologic view]. AB - Usually, pain is caused by the excitation of nociceptors or of nociceptive afferent fibers. The responsiveness of the nociceptors can be modulated by endogenously released algesic substances or local hormones. Lesioned and regenerating nerve fibers show enhanced excitability and spontaneous activity. Motor and sympathetic reflexes sometimes have an excitatory action on nociceptors, a positive feedback which often is essential in chronic pain states such as the sympathetic reflex dystrophies. Proper diagnosis of these influences on nociceptor excitation is crucial if therapeutic interference is to be rendered possible at the peripheral site of encoding of pain stimuli. No pain center exists in the central nervous system. The cognitive, affective, motor and vegetative dimensions of pain perception and pain behaviour are based on a complex interaction of several brain systems. An important feature of brain function in relation to pain is the fact that nervous information related to pain can be modulated in the central nervous system. Various inhibitory modulations have recently been described which may be used for pain therapy. Inhibition of spinal neurons, and analgesia, can be produced by stimulation of brain stem structures. Pharmacologically, pain inhibitory systems may involve serotonin and endogenous opioids as transmitter or modulator substances. For pain therapy, pain inhibitory systems may be activated e.g. by morphine and ther analgesic drugs, focal brain stimulation, various means of somatosensory afferent stimulation and by psychological influences such as stress. PMID- 6276971 TI - [Endorphins and the occurrence of pain]. PMID- 6276972 TI - [Clinical and therapeutic study of a group of 217 patients with intestinal giardiasis and amebiasis]. AB - In 15 months of the years 1979-1980, 158 patients with giardiasis and 69 patients with intestinal amebiasis were diagnosed and treated in the outpatient department of the Swiss Tropical Institute. Repeated examination of merthiolate-iodine formol-fixed stools was the method of choice for detecting intestinal protozoa. Parasitological investigation of duodenal fluid (Enterotest) was less effective. Fewer than 30% of the cases were symptomatic, usually mentioning diarrhea (73 85%) or abdominal discomfort (20-44%). Treatment was by ornidazole single dose (2 g in adults; 40 mg/kg in children) for asymptomatic cases and a five-day treatment schedule (2 X 500 mg/day in adults, 20 mg/day in children) for symptomatic cases. In amebiasis a luminal amebicide (diloxanide furoate) was prescribed in all instances after the first stool examination. Parasitological cure rates were excellent (95%) in all cases of giardiasis but only 59 and 65% respectively in cases of asymptomatic and symptomatic amebiasis. Drug tolerance was excellent. Ornidazole is the drug of choice for all forms of giardiasis, even in a single dose. As a luminal amebicide, however, its efficacy is not superior to other nitroimidazole derivatives. PMID- 6276973 TI - [Treatment of urinary infection with a single dose of co-trimoxazole compared with a single dose of amoxicillin and a placebo]. PMID- 6276974 TI - The structure of chromatin. PMID- 6276976 TI - Viral receptors on isolated murine and human ependymal cells. AB - Viruses that infect ependyma cause ependymitis in humans and hydrocephalus in experimental animals. We report that reovirus type 1 (which induces hydrocephalus in mice) binds to the surface of isolated human and murine ciliated ependymal cells. With the use of recombinant viral clones, the binding property was mapped to the type 1 viral hemagglutinin, which also determines in vivo the affinity of reovirus type 1 for ependyma. Mumps virus, measles virus, parainfluenza type 3, and herpes simplex virus type 1 bind to murine ependyma cells, whereas reovirus type 3, herpes simplex virus type 2, and poliovirus type 2 do not. PMID- 6276975 TI - Expression of bovine leukemia virus genome is blocked by a nonimmunoglobulin protein in plasma from infected cattle. AB - Plasma of cattle infected with bovine leukemia virus contains a soluble factor that blocks the expression of the viral genome in cultured lymphocytes. The blocking factor is not present in plasma of bovine leukemia virus-free cattle or of cattle infected with common bovine viruses. Blocking of bovine leukemia virus expression by the plasma factor is reversible, and seems to be mediated by a nonimmunoglobulin protein molecule. PMID- 6276977 TI - Corticotropin-releasing activity of alpha-melanotropin. AB - Synthetic alpha-melanotropin stimulated the release of immunoreactive adrenocorticotropin from primary cultures of rat anterior pituitary cells. The effect of the alpha-melanotropin was dose-dependent. Cells incubated with synthetic arginine-vasopressin and alpha-melanotropin simultaneously produced an amount of adrenocorticotropin that was greater than the sum of the amount that the cells produced in response to each peptide added separately. Other peptides structurally similar to alpha-melanotropin, such as, beta-, gamma 1-, gamma 2-, and gamma 3-melanotropin, were also tested for adrenocorticotropin-releasing activity. Only the gamma 3-melanotropin demonstrated a statistically significant effect. A vasopressin preparation (Pitressin, Parke-Davis) purified from posterior pituitaries and previously shown to contain some alpha-melanotropin was much more potent in releasing adrenocorticotropin than the synthetic vasopressin. PMID- 6276978 TI - DNA strand breakage in human leukocytes exposed to a tumor promoter, phorbol myristate acetate. PMID- 6276979 TI - [Diuretics and uric acid (author's transl)]. AB - Long-term diuretic therapy often increases serum uric acid. The renal metabolism of uric acid is recalled and the factors which lead to diuretic-induced hyperuricemia are discussed. Fluid and sodium loss appears to be the main cause of increased serum uric acid. The authors' experience with management of severe hypertension is consistent with this theory. The effect of tienilic acid which lowers serum uric acid levels is discussed. PMID- 6276980 TI - [Hepatitis, cholestasis and amineptine (author's transl)]. AB - Eight cases of hepatitis induced by amineptine chlorhydrate, a new tricyclic antidepressant are reported. Jaundice and/or biological changes occurred 16 to 75 days after the onset of treatment and with a total oral dose of 3,8 to 26 grams. Biologically, cholestasis and cytolysis have been observed simultaneously in 4 cases, cholestasis alone in 2 cases, and prevalent cytolysis in 2 others. After withdrawal of amineptine, clinical improvement was achieved within 10 to 21 days, whereas biological changes of hepatic function tests lasted for as long as 3 to 12 weeks. Physiopathology of such disorders is not clear : toxicity, enzyme induction, immunoallergy, may be considered as possible mechanisms. PMID- 6276981 TI - [Hemorrhage in liver cirrhosis : new suggestions (author's transl)]. AB - Current management of hemorrhage in cirrhotic patients is disappointing, probably because it deals only with the portal hypertension, while the coagulation disorders are neglected. Some new suggestions can be made : 1) Hemorrhage originates in coagulation disorders. The mechanical lesion of the mucosa is only the opportunity for these disorders to become apparent. The lesion may be : infrequently, a ruptured esophageal varix or a gastroduodenal peptic ulcer ; a lesion of the cardia (hiatal hernia, reflux, esophagitis, minimal traumatic tears) ; a gastric anomaly (hemorrhagic gastritis, superficial ulcerations, petechiae) ; in some cases no mucosal lesion is apparent. 2) Any widespread liver disease results in lasting hypercoagulability which is responsible for : permanent lysis, consumption, DIC. The spleen is responsible for the functional alteration of the platelets. Splenectomy is followed by permanent recovery. 3) Changes involving the platelets are responsible for most hemorrhages. Thrombopenia and severe anomalies of platelet aggregation are common findings in liver cirrhosis. Further deterioration can be induced by acetylsalicylic acid, especially if it is absorbed after an immoderate ingestion of alcohol. Emergency treatment consists in platelet transfusions. 4) Stasis in the portal system may, however, result in permanent activation of coagulation. 5) Cirrhosis results in chronic hypercoagulability and severe platelet deterioration. Any stress involving coagulation mechanisms may therefore induce hemorrhage : infection, acetyl salicylic acid, respiratory distress, estrogens, massive transfusion. It is always dangerous to "feed" consumption or to restrain lysis. 6) Coagulation tests should be performed rapidly, in order to evaluate hypercoagulability, consumption, lysis, and evidence of DIC ; FDP can probably be responsible for inflammatory changes in the liver and spleen. 8) Coagulation disorders are permanent since the hepatic alterations are irreversible. PMID- 6276982 TI - [Acquired renal cystic disease in patient on long-term hemodialysis (author's transl)]. AB - We report two cases of acquired cystic disease of the kidneys in patients on long term hemodialysis. The gross appearance of these polycystic kidneys can be distinguished from the congenital adult type by their normal size and weight and by the small range of their cysts. We found some similarity between these numerous cysts and the simple retention cysts. This polycystic disease could concern about 30 % of the whole patients on long-term hemodialysis ; the mechanisms of its development are unknown. Clinically, this lesion is mostly asymptomatic but could sometimes be discovered by intra or perirenal haemorrhage. PMID- 6276983 TI - [Evaluation of venotropic drugs by venous gas plethysmography. A study of procyanidolic oligomers (author's transl)]. AB - Venous tone in the lower limbs can be measured by venous occlusion gas plethysmography before and after giving venotropic drugs. Four groups, each of which included ten patients with widespread varicose veins, were given different medications. Each patient was studied by plethysmography during the first five hours following drug ingestion. In untreated controls, venous tone decreased during rest. A reference drug was uneffective. Increase in venous tone induced by 150 mg of procyanidolic oligomers was comparable to that which followed ingestion of a high dose of hamamelis-hydrastis mixture. PMID- 6276984 TI - [Epiperineural nerve sutures and neurilemmatic sutures. Comparative clinical results about 109 cases (author's transl)]. AB - During the last ten years, 109 medical cases of traumatic hand nerve lesions have been observed, implicating 78 big nerves (median 35, cubital 42, radial 1) and 37 little nerves (median 35, cubital 42, radial 1) and 37 little nerves (interosseous, collateral, digital). 81 "mixte" epiperineural sutures have been realized (according to a technical proceeding purposed by one of the authors since 1970, and practised with the only aid of a simple optical increase by dissection's magnifying-glasses) : In the same time, 38 neurilemmatic sutures have been seen at second hand and their clinical results noted. 64+ of the global results are very good or "good" concerning the "mixte" epiperineural sutures against 34% for the neurilemmatic sutures. To note : the best quality of the results for the median nerve as well for the primitive sutures, and unfavourable influence of the associated lesions. PMID- 6276985 TI - [Tiapride : a clinical study in a department of internal medicine (author's transl)]. AB - Effectiveness and tolerance of tiapride were studied in twenty patients in a department of internal medicine. Tiapride was effective against acute manifestations of alcoholism and against agitation, especially in the elderly. There were no serious side-effects, particularly on hemodynamics and vigilance. At the dosage used in our study the analgesic effect of tiapride was both unreliable and transient. PMID- 6276986 TI - [Involvement of the common bile duct in Hodgkin's disease : report of three cases (author's transl)]. AB - The authors report 3 cases of extra-hepatic obstruction of the common bile duct in Hodgkin's disease, in one case due to lymph nodes compressing the hepatic pedicle, in the second case due to infiltration of the common bile duct alone, in the third case by a large Hodgkin tumour of the head of the pancreas. Although emphasizing the rareness of these cases of jaundice in Hodgkin's disease, they emphasize the necessity of exploring these patients in order to make a precise assessment of the lesions and thus choose the best treatment. PMID- 6276987 TI - [Immediate arterial hypertension post-kidney traumatism (author's transl)]. AB - Arterial hypertension is uncommon shortly after renal trauma. It is due to peri renal hematoma following mild lesions. Transient, moderate or severe, this hypertension is hyperreninemic. Only medical treatment is justified. PMID- 6276988 TI - [Bronchocentric granulomatosis (author's transl)]. AB - A case report and a review of this recently described entity are presented. A feature of interest in our case is the presence of Aspergillus fumigatus at the pulmonary needle biopsy as well as in the surgical specimen. This finding is described for the first time in a non asthmatic patient. PMID- 6276989 TI - [Treatment of non seminomatous testicular tumors (author's transl)]. AB - Progress in the diagnosis and treatment of NSTT have been registered during the last years. Tumor markers have permitted to lower the percentage of staging error ; nevertheless they have their limits and a return to normal of tumor markers do not mean obligatorily the disparition of a viable tumor. The treatment of NSTT is an emergency : lymphadenectomy is now, for most of authors limited and guided by extemporaneal exploration ; radiotherapy has a complementary target of detecting agent ; chemotherapy (particularly since CDDP was introduced) is very efficient in disseminated forms of disease giving long term remission in 50 to 70 % of cases. Actual results give, for next years, the hope of eradication of NSST in a high number of cases. PMID- 6276990 TI - [Laennec and his times]. PMID- 6276991 TI - [Dr. Clemenceau]. PMID- 6276992 TI - [Pulmonary involvement in chronic lymphocytic leukemia. Diagnostic value of trans bronchial biopsy (author's transl)]. AB - Pulmonary parenchymatous involvement in a patient with chronic lymphocytic leukemia (CLL) was demonstrated by trans-bronchial pulmonary biopsy using a flexible endoscope. This procedure is clearly of value in similar cases when clinical and roentgenological examinations are insufficient. The observed cases provides the opportunity for a brief chronological review of the literature. Advances in the classification of CLL, which follow the technological improvements, are described as well as their bearing on management and prognosis of this condition. PMID- 6276993 TI - [Hodgkin's disease and idiopathic cardiomyopathy (author's transl)]. AB - The authors report the case of a 36 years old man who has been successfully treated, ten years ago, with radiation therapy for stage II Hodgkin's disease, with sub-diaphragmatic involvement. The clinical course was unhappily marked by the development of progressive congestive heart failure which was related to an "idiopathic cardiomyopathy" anatomically confirmed. A literature review does not permit to find such another case. It was not possible to conclude if this association was fortuitous or not. But if could be compared to the increased risk of post-transplant lymphomas in patients with idiopathic cardiomyopathy. PMID- 6276994 TI - [Intractable menopausal disorders cured by veralipride (author's transl)]. AB - We studied a case of surgical menopause in a hysterical patient. Castration was followed by depression with sexual disorders, somatic complaints, and specific menopausal manifestations, mainly hot flushes. Hysterectomy had been done a year earlier. Since then the patient had been admitted twice to a psychiatric ward following attempted suicide. Veralipride was given for twenty days. Noticeable improvement was recorded. Therapy was then discontinued for ten days. Symptoms recurred and long-lasting therapy was decided on. Subsequently, sustained overall improvement with disappearance of specific menopausal disorders, were recorded. PMID- 6276995 TI - [Retroperitoneal malignant xanthogranuloma (author's transl)]. AB - A rare retroperitoneal malignant xanthogranuloma is described. The patient developed malignant bone lesions followed by brain localization, and died three years later. At autopsy a very large cystic retroperitoneal tumor with osteolytic lesion of sacrum was found. A review of cases diagnosed as retroperitoneal xanthogranuloma indicates that this lesion seems to be a particular form of the wide group of malignant fibrous histiocytoma with xanthomatous and inflammatory cells. Moreover it could be a primary malignant fibrous histiocytoma of bone. Furthermore, difficulties of differential diagnosis of these tumors are discussed mainly with histiocytosis X. PMID- 6276996 TI - [Atypical osteodystrophy of visceral origin (author's transl)]. AB - The authors report a case of atypical osteodystrophy of visceral origin (uretero sigmoid by-pass for bladder exstrophy) which was characterized by the association of osteocondensation and looser-Milkman lines. They emphasize the rareness of osteodystrophy after uro-digestive by-pass and the exceptional character of the osteocondensation. PMID- 6276997 TI - [Continuous predetermined insulin infusion in insulin-dependent diabetes mellitus (author's transl)]. AB - At the present, prognosis of insulin-dependent diabetes depends on it's degenerative complications. Many studies have confirmed that the incidence of severe degenerative complications of early onset parallels the degree of metabolic unbalance. Unfortunately, in many diabetic patients satisfactory metabolic control is difficult to ensure because of the irregular absorption of insulin injected discontinuously in subcutaneous tissues. This is true even under strict diet conditions. Continuous insulin infusion at a predetermined dosage was therefore suggested. Data on basic requirements is provided by an artificial pancreas. Satisfactory results have already been reported with intravenous and subcutaneous routes. Intramuscular and intraperitoneal routes are at present under trial. The use of portable pumps facilitates these techniques. The insulin dosage must be correctly determined and adjusted to the patient's diet and activity. In brittle diabetes, these methods can provide, at least temporarily, a satisfactory metabolic balance, and useful data for adjusting further therapy. PMID- 6276998 TI - [Severe gastrointestinal manifestations of Henoch-Schonlein purpura in adults (author's transl)]. AB - Two cases of Henoch-Schonlein purpura in adults are reported. Both patients presented with severe digestive symptoms suggesting malabsorption or exudative enteropathy. Roentgenological examinations showed marked anomalies of the small intestine as well as changes suggestive in intra-mural hematoma located in the stomach in one case and in the duodenum in the other. Hypoalbuminemia occurred in both patients during the course of the disease and was highly suggestive of protein-loosing enteropathy. Disseminated intravascular coagulation was seen in one patient. Both patients recovered from their gastrointestinal disease. Roentgenologic changes disappeared in one case; in the other, marked compression and distension of the duodenum were still visible six months after onset. PMID- 6276999 TI - [Carbon dioxide laser in the treatment of premalignant lesions of the cervix (author's transl)]. AB - Cervical dysplasia is classified in various groups according to the extent of the epithelial anomalies. Severe dysplasia, in which the entire depth of the epithelium is diseased, is assimilated to the situ carcinoma. The diagnosis is established by cytology and above all by colposcopy which enables to guide the biopsy. Since all dysplasias, whether mild, moderate or severe, may lead to cancer, their treatment is always warranted. The carbon dioxide laser appears to be superior to the other methods of local therapy. Exocervical dysplasia requires ablation for histological examination, followed by a 7 mm-deep tissue destruction. The carbon dioxide laser also allows conization for endocervical dysplasias. A perfect knowledge of colposcopy is indispensable for this therapeutic procedure. PMID- 6277000 TI - [A new observation of an H-Y antigen positive 46, XX male (author's transl)]. AB - A new serologic test for detecting the H-Y antigen by indirect fluorescence performed on purified lymphocytes was developed. It allowed us to demonstrate H-Y antigen in an 46, XX male. The presence of this antigen explains why the primitive gonad underwent testicular differentiation. Several mechanisms could account for the presence of H-Y antigen. Mosaicism or translocation of an X on a Y or an autosome may prevent detection of a Y chromosome. Alternatively, a Y chromosome may have disappeared during embryogenesis, The last hypothesis suggests that an autosome may carry a structural gene coding for H-Y antigen; in this case thye Y chromosome would act only as an accessory regulating gene. PMID- 6277001 TI - [Pleuropulmonary manifestations in amebiasis. Sixty-one cases seen in Abidjan over a three-year period (author's transl)]. AB - Sixty-one observations of pleuropulmonary amebiasis are reported with a detailed analysis of radioclinical features In 13.1% of the cases there were no clinical signs of associated hepatic involvement. Diagnostic difficulties were encountered in 42.6% of the cases which were suggestive of bacterial pleuropulmonary disease, tuberculosis or carcinoma. Serologic tests for amebiasis, which were positive in 95% of the cases, are very helpful diagnostic tools. Metronidazole and its derivatives were used in 80% of the cases with satisfactory results: death occurred in 3.5% of the cases and good short-term results in 84.6%. Sequelae were mainly observed after amebic empyema which resulted in chronic pleural disorders in 27.7% of the cases. PMID- 6277002 TI - [Myocardiopathies secondary to cardiovascular diseases: systolic strain and diastolic overload (author's transl)]. AB - All cardiovascular malformations compromise ventricular function and lead to alterations in the myocardial structure. Most of these are adaptive reactions. However, in some cases, this reaction overreaches its object and leads to an authentic myocardiopathy. This myocardiopathy is usually hypertrophic in obstacles which result in systolic overload of the left ventricle (coarctation peri-orificial stenosis of the aorta, systemic hypertension) or of the right ventricle (pulmonic stenosis), or congestive in diastolic overload by left-to right shunts or valvular insufficiency. These myocardiopathies may prevent recognition of the signs of the underlying heart disease thereby leading to diagnostic errors and delay in therapy. They may constitute the main factor of an operative decision. PMID- 6277003 TI - [Prognostic features of upper gastro-intestinal hemorrhages in adult over 65 (author's transl)]. AB - Some prognostic features out of 57 cases at upper gastrointestinal hemorrhage, in people more than 65 years old, are reported. The age has no significance but after 70, gastroduodenal ulcers represent the more frequent etiology, with a very particular severity in cases of chronic and acute gastric ulceration. The mortality is still high, mainly due to decompensation of pre-existent renal, cardio-vascular or pulmonary taints, frequent in aged people. PMID- 6277004 TI - [Endocarditis due to commensal Neisseria (author's transl)]. AB - Three cases of endocarditis due to Neisseria mucosa are reported. The pathogenic role of these commensal microorganisms is reviewed. A survey of the literature found a dozen cases of endocarditis due to commensal Neisseria before the antibiotic era, and fourteen more since then (not including our three observations). Some of distinctive features can be delineated from the study of cases seen during the second period. Patients mean age is fairly low (34 years), incidence of cutaneous manifestations is high, and vascular complications (mainly embolism) are common (52, 1% of the cases). Overall prognosis is good and the recovery rate (82 %) is similar to that seen in non-D Streptococcus endocarditis. PMID- 6277005 TI - [Bacteriologic and pharmacodynamic basis for treating bacterial meningitis (author's transl)]. AB - In managing bacterial meningitis, the essential rule, stated by Chabbert, is to achieved antibiotic concentrations in the CSF which exceed the minimal bactericidal concentration (MBC) Several factors must be considered, related either to the microorganism itself (virulence, sensitivity, density of the cultured sample, resistance to metabolites, inactivation of the antibiotic by purulent CSF) or to the patient (initial site of infection, immunologic deficiency, csf clearance). As to the antibiotic liposolubility, free plasma fraction, and route of administration, are the factors which affect it's passage towards the CSF. PMID- 6277006 TI - [Brucella melitensis endocarditis. Clinical and immunological features (author's transl)]. AB - Five cases of Brucella melitensis endocarditis are reported. Common features included subacute course, enhanced humoral immunity, and deficient cellular immunity. Appropriate antibiotic therapy led to recovery in all cases. In two patients, valvular replacement was necessary. PMID- 6277007 TI - [Antibiotic treatment of brucellosis (author's transl)]. AB - Tetracyclines are remarkably effective in brucellosis and give consistently good results. However, recurrences have been reported even after correct management. Combination therapy with tetracycline and streptomycin, advocated by the FAO/WHO, has improved results without giving complete satisfaction. Association of tetracycline with rifampicin suggested on sound theoretical grounds, has given promising results. PMID- 6277008 TI - [Urethral complications of severe fractures of the pelvis. (author's transl)]. AB - Between 1975 and 1979, 282 fractures of the pelvis were treated in Centre Hospitalier Universitaire Vaudois' sixty-eight of these patients sustained severe fractures, with two or more rupture of the pelvic ring. Twenty-eight patients presented with hematuria (microscopic or macroscopic) and five others with urethral bleeding. Urethral bleeding is even more serious; in the present study, it was always associated with urethral rupture. These cases are discussed; mechanism, diagnosis and treatment of these injuries are described. PMID- 6277009 TI - [Spiramycin ; therapeutic value in humans. (author's transl)]. AB - Spiramycin gives high tissue levels and has very-few side-effects. It is one of the best antibiotics in infections of the upper and lower respiratory tract and of the oral cavity. It is probably one of the most effective antibiotics against chlamydia. Spiramycin is useful when toxoplasmosis occurs during pregnancy or results in severe diseases in non-immunodepressed patients. For the reasons listed above, we believe that spiramycin has been underestimated. This is the result of assessing its value according to its rather high MIC rather than to it's effectiveness against infection in animals and humans. PMID- 6277010 TI - [Urogenital tuberculosis : current status and future perspectives (author's transl)]. AB - Results of a retrospective study of 72 patients with urogenital tuberculosis admitted to Hopital Cochin Paris, from 1970 to 1980, were compared with those of studies conducted since 1945. Though there has been a marked reduction in the number of cases of tuberculosis in absolute terms, morbidity has not altered as a nephrectomy was performed in 44 p. cent of cases both in the period 1970 to 1980, and the year 1950. There has, however, been a noticeable increase in the use of plastic surgery. Future perspectives involve both the medical and surgical fields, in the former leading to a shorter period (9 months) of treatment, and in the latter by the development of new endoscopic procedures: modelling ureteral sounds, and hydrostatic bladder distention. It is reasonable to assume that surgery will be required less often in the future, medical treatment involving endoscopic procedures being sufficient to treat the majority of urogenital tuberculosis cases. PMID- 6277011 TI - [Corynebacterium endocarditis. Diagnostic difficulties (author's transl)]. AB - The authors report a case of endocarditis due to Corynebacterium diphtheriae. THe bacteriological study is difficult since this micro-organism can be undistinguishable from a saprophytic diphtheroid. The observed case is compared to previously published observations. All have in common the great severity of the illness, and the high mortality rate. Despite the clinical symptoms and the cerebrospinal fluid (CSF) abnormalities, meningitic involvement could not be demonstrated. PMID- 6277013 TI - Horner's syndrome due to congenital cytomegalovirus infection. PMID- 6277012 TI - [Pneumococcal septicemia: 145 cases (author's transl)]. AB - Over a six year period, 145 pneumococcal septicemias were seen at the Tourcoing Hospital. This represents 0.39% of all hospitalized patients and 19.8% of all septicemias seen over the same period. A mortality rate of 38.6% was recorded. Prognostic factors were analyzed. The mortality rate was significantly higher in women (46.5%), in patients admitted for a reason unrelated to the septicemia (50.8%), and in patients with septic shock (81.5%) which was frequent (22.8%). An underlying poor general condition was present in 88.2% of the patients. This confirms the high risk of pneumococcal infections in some patients who should be protected by vaccination. PMID- 6277014 TI - Cutaneous and oral changes as the only manifestations of the glucagonoma syndrome. PMID- 6277016 TI - The specific binding of the thyroid hormones to matrix isolated from rat liver nuclei. AB - Specific binding sites for the thyroid hormones have been demonstrated in the liver nuclear matrix, a structural framework of the nucleus. When labelled 3,5,3' tri-iodo-L-thyronine ([125I]T3) is injected into rats, 5% of the total nucleus bound T3 is bound to the matrix after 1 hour. However, when either isolated nuclei or isolated nuclear matrices were incubated with [125I]T3 in vitro, a 3- to 7-fold greater number of specific T3 binding sites were revealed in the nuclear matrix. The properties of the matrix-associated thyroid hormone binding sites were investigated in vitro. These binding sites showed limited capacity and high affinity for T3; the equilibrium association constant (Ka) was 1,3 x 10(9) M 1 and the binding capacity was 20,2 fmol T3 per 100 microgram matrix protein. PMID- 6277017 TI - Tumours of the heart. A study of 89 cases. AB - Eight-nine patients with heart tumours (76 metastatic and 13 primary) were studied. Metastatic tumours were usually clinically silent and most often came from lung carcinomas, lymphomas, malignant melanomas, and gastric and breast cancers. The primary tumours consisted of 11 myxomas, 1 fibrous histiocytoma and 1 haemangioma. PMID- 6277015 TI - [Use of a contraceptive suppository as chemoprophylaxis against sexually transmitted diseases]. PMID- 6277018 TI - Ranitidine heals duodenal ulcers. AB - A double-blind placebo-controlled study of ranitidine 150 mg given twice daily for 4 weeks was carried out in 80 ambulant patients with endoscopically proven duodenal ulceration. Fifteen patients were withdrawn because of poor compliance or default. Of the remaining 65 patients, endoscopic healing (with or without residual erosion) was noted in 28 of the 34 ranitidine-treated patients (82%) and in 14 of the 31 placebo-treated patients (45%) (P less than 0.005). Twenty-three patients whose ulcers were unhealed after 4 weeks of ranitidine therapy or placebo were then given ranitidine for a further 4-week period. Four of these patients were withdrawn, but ulcer healing was achieved in 16 of the remaining 19 patients (4 out of 5 (80%) initially treated with ranitidine and 12 of 14 (86%) initially treated with placebo). No significant clinical, biochemical or haematological adverse reactions were noted in either treatment group. It is concluded that ranitidine is a safe and effective therapy in patients with duodenal ulceration and that initial unsuccessful treatment with placebo does not appear to influence the subsequent response to ranitidine. PMID- 6277019 TI - Ranitidine in the treatment of gastric ulceration. AB - Seventy-two patients with endoscopically proven gastric ulcers were entered into a prospective controlled trial to assess the efficacy of ranitidine and cimetidine in ulcer healing. All patients were biopsied on entry and at subsequent endoscopies. After exclusion of 7 patients during the first month of treatment, the remaining 65 patients, 47 males and 18 females, mean age 48.2 +/ 1.5 years, at 1 month had a healing rate of 47% and 52% respectively. The non healers continued their treatment for a further 4 weeks. This increased the healing rate to 77% and 76% respectively. If the defaulters and poor compliers are withdrawn the healing rate rises to 58% and 57% at 4 weeks and to 91% and 79% at 8 weeks respectively. There was no significant difference between the two groups as regarded initial ulcer size and severity of dyspepsia. Antacid tablet consumption during the study was comparable. The initial size of the ulcers which failed to heal after 4 weeks of treatment tended to be larger than those which healed (P less than 0.05), but smoking did not appear to influence ulcer healing. No obvious side-effects or evidence of dysplasia were found. The study shows that ranitidine is at least as effective as cimetidine in gastric ulcer healing. PMID- 6277020 TI - Guar biscuits in the diabetic diet. AB - A new type of guar-containing biscuit has been developed and incorporated into the diabetic diet in both short- and medium-term studies. It has been found to be effective in reducing the postprandial rise in the blood glucose level and in improving glycaemic control, as shown by reduced fasting blood glucose values and decreased 24-hour urinary glucose excretion. This form of guar has proved to be palatable and acceptable to patients. It was effective in smaller quantities than have previously been tested and may prove a valuable adjunct in the treatment of diabetes. PMID- 6277021 TI - Sexually transmitted diseases in homosexuals: focusing the attack. PMID- 6277022 TI - Genital warts. AB - Human papillomaviruses have not been propagated in tissue culture, so they cannot be studied by the procedures of classical virology. Recently, DNA hydridization techniques and restriction enzyme analysis have revealed that multiple strains of human papillomaviruses exist. It is now accepted that genital warts are a sexually transmitted disease; other routes of infection are possible, but unusual. An association between maternal warts during pregnancy and the development of vulval warts and laryngeal papillomatosis in infants and young children has been shown. The epidemiology of anal warts in homosexual men is not well understood. The flat condyloma, visible on colposcopy, is a human papillomavirus-containing lesion with a wart-like histology. It is a common condition which in the past may have been mistaken for dysplasia. Flat condylomas may regress spontaneously, or develop into condylomata acuminata. A link between these lesions and some cases of cervical carcinoma has been suggested. PMID- 6277023 TI - Sexually transmitted infections and cervical atypia. AB - Epidemiologic studies have long associated cervical cancer and dysplasia with sexual activity. In the past two decades an abundance of biological and epidemiologic data have indicated a relationship between herpes simplex virus type 2 (HSV-2) and these conditions. Unfortunately, the strength of the evidence (albeit circumstantial) has led many researchers to focus exclusively on the putative role of HSV-2 as the carcinogen. At the same time, many studies have shown that cervical atypia, if it is induced by HSV-2, must be a very rare outcome resulting from a very common exposure. On the basis of the assumption that all forms of dysplasia need not have a common cause and the likelihood that other sexually transmitted cervical pathogens (e.g., Chlamydia trachomatis, trichomonas vaginalis, papillomavirus, and perhaps cytomegalovirus) may be able to induce atypia and perhaps interact with HSV-2 to produce that outcome, the plea is made for inclusion of other sexually transmitted agents in prospective studies on cervical atypia. PMID- 6277024 TI - The role of estrogen receptor status in predicting the response of carcinoma of the breast to adjuvant chemotherapy. AB - This study was undertaken to determine if there was any correlation between the estrogen receptor status and the response of patients with operable carcinoma of the breast to adjuvant chemotherapy. Seventy-seven patients with metastases to the axillary lymph nodes who were receiving adjuvant chemotherapy following a modified radical mastectomy were studied. During the period of study, 12 of 34 estrogen receptor-positive tumors recurred, with a mean disease-free interval of 17.8 months, and 19 of 43 estrogen receptor-negative tumors recurred, with a disease-free interval of 15.2 months. Statistical analysis of recurrence at two years showed no difference between these two groups. Only one patient of 18 in the premenopausal group of women with estrogen receptor-positive tumors, however, had a recurrence compared with 11 of 24 patients with estrogen receptor-negative tumors. This difference was highly statistically significant, p greater than 0.0005. We have concluded that the estrogen receptor status is not a reliable indicator of response to adjuvant chemotherapy for all patients. Our data do indicate a positive correlation between response to adjuvant chemotherapy and estrogen receptor status in the premenopausal group of patients and suggest that adjuvant chemotherapy in the premenopausal group of patients may be exerting some of its effect by ovarian suppression rather than by solely cytotoxic effect on cancer cells of the breast. PMID- 6277025 TI - New embolization method using estrogen for dural arteriovenous malformation and meningioma. AB - For the treatment of dural arteriovenous malformations, conjugated estrogens were infused continuously through a cannula inserted into the external carotid artery. In 6 or 8 patients, therapeutic effects such as total or partial disappearance of the nidus were observed. The same method was employed to occlude the feeders of meningiomas in 3 patients to reduce bleeding during the operations. The histological findings from the surgically extirpated meningioma are described. PMID- 6277026 TI - Detection of experimental brain tumor in rat by computed tomography. AB - The feasibility of the use of computerized tomographic (CT) scanning for detecting an experimental brain tumor in rats was evaluated. Tumors were induced in newborn rats by intracerebral inoculation of Rous sarcoma virus. At varying times contrast-enhanced scans were obtained on an Ohio Nuclear 2010 scanner, the brains examined, and the findings at autopsy correlated with the CT findings. Five tumors were demonstrated by CT scans and their presence confirmed at autopsy. Four small tumors were not detected by CT scans. In 1 animal, multiple tumors were demonstrated by a CT scan, and in another a large cyst was correctly diagnosed. It is concluded that the technology currently available can be used to identify some tumor-bearing animals and should be useful for following the change in size of tumors in response to therapy. Further improvements in scanner resolution should make this technique more useful for animal research. PMID- 6277027 TI - Multicentricity and bilaterality in invasive breast carcinoma. AB - Multicentricity and bilaterality are well-established characteristics of breast carcinoma, but little is known about the relationship of these variables with each other. This question was explored by analyzing the data pertaining to 880 women with invasive breast carcinoma. Patients with multicentric carcinoma had bilateral disease more often than those whose carcinoma was apparently limited to a single quadrant. Among women who had lobular carcinoma in situ coexisting with infiltrating duct carcinoma or infiltrating lobular carcinoma, bilaterality and multicentricity were significantly more common than they were among patients whose only lesion was infiltrating duct or medullary cancer. Other variables associated with bilaterality and multicentricity were degree of ductal differentiation, tumor size, nodal status, type of tumor margin, intensity of lymphoid infiltrate, and menstrual status. Age at diagnosis and estrogen receptors were related to bilaterality but not to multicentricity. The following variables proved to be unrelated to bilaterality and multicentricity: family history of breast carcinoma, height, weight, and parity. The data obtained in this study tend to support a conclusion that multicentricity and bilaterality are manifestations of similar factors involved in the neoplastic transformation of mammary gland epithelium leading to the development of breast cancer. PMID- 6277029 TI - [Visceral leishmaniasis after a Mediterranean trip]. PMID- 6277028 TI - Psoas weakness and femoral neuropathy: neglected signs of retroperitoneal hemorrhage from ruptured aneurysm. AB - Femoral neuropathy may occur with aortic aneurysm more commonly than reports in the literature indicate. The combination of an aortic aneurysm and femoral neuropathy indicates rupture. The presence of abdominal pain and neuropathy should suggest ruptured aneurysm and exclude other commonly considered diagnoses. Preoperative recognition of femoral neuropathy provides the opportunity for intraoperative nerve decompression and assures the surgeon that the operation itself was not causative. The literature is reviewed, and the preoperative occurrence of femoral neuropathy in two patients with ruptured aortic aneurysms is described. PMID- 6277031 TI - Angiotensin converting enzyme in bronchoalveolar lavage fluid in pulmonary sarcoidosis. AB - Alveolar angiotensin converting enzyme (ACE) and serum ACE were measured simultaneously in 16 patients with histologically confirmed sarcoidosis and in 16 control subjects. Although alveolar ACE was abnormally elevated in all 15 cases of active sarcoidosis, serum ACE was not. No correlations were found between radiographic staging of pulmonary sarcoidosis and the levels of these enzymes. There was a clear correlation, however, between the levels of alveolar ACE and counts made on bronchoalveolar lavage fluid. This correlation was closer than that existing between serum ACE and bronchoalveolar lavaged lymphocytes. It is suggested that alveolar ACE is an additional biological marker of pulmonary sarcoidosis which is possibly more sensitive than serum ACE. PMID- 6277030 TI - Mast cells and basophils: effector cells of inflammatory disorders in the lung. PMID- 6277032 TI - Alveolar response to injury. PMID- 6277034 TI - Valid evaluation of plasma catecholamines. PMID- 6277033 TI - Plasma cyclic nucleotide levels in exercise-induced asthma. AB - It is known that sympatho-adrenal control of airways is increased in asthma since beta blockade can cause severe bronchoconstriction in asthmatic individuals. It has not been established whether an altered catecholamine response to exercise plays any part in the production of the common symptom of exercise-induced asthma (EIA). We have investigated this indirectly by measuring arterial plasma cyclic nucleotide levels in 10 subjects with EIA and five normal subjects. Cyclic AMP, which in this context reflects beta stimulation, rose significantly by 25.4% in the normal subjects during exercise, while there was no significant change during or after exercise (less than 5%) in the asthmatic subjects. Cyclic GMP rose significantly after exercise in the asthmatic subjects. Six normal subjects repeated the protocol before and after inhalation of salbutamol aerosol, 1600 microgram daily for 18 days. This did not reduce the cAMP response to exercise, and we conclude that the diminished cAMP response of the asthmatic subjects was not caused by their medication. The results may indicate either impaired catecholamine production or endogenous beta receptor hyporesponsiveness in some asthmatic subjects and this may contribute to the development of EIA. PMID- 6277035 TI - Effect of flunarizine on the human red cell shape changes and calcium deposition induced by A 23187. PMID- 6277036 TI - Prostaglandin precursors in platelet phosphoglycerides following acute myocardial infarction. PMID- 6277037 TI - Characterization of antibodies reacting with husband's lymphocytes in sera from patients with trophoblastic malignancies. AB - Lymphocytotoxic antibodies reacting with husband's lymphocytes were demonstrated in 24 sera from women with trophoblastic neoplasia studied before any chemotherapy. These antibodies exhibited a maximal cytotoxic activity at +4 degrees C. They reacted mainly with B lymphocytes as assessed by microlymphocytotoxicity assays using B- or T-cell enriched subpopulations and T- or B-cell lines as targets. They could not be absorbed out with platelets or erythrocytes, and did not react with autologous lymphocytes. In one double labelling immunofluorescence experiment, these antibodies could be stained by rhodamine-conjugated Fab'2 anti-mu fragments and thus appeared to belong to the IgM class. They were shown to react mainly with surface Ig (SIg) bearing lymphocytes, plus a minor SIg negative subset. Studies with panels of allogeneic normal B cells and of HLA homozygous B-cell lines showed that the target antigen(s) recognized by these antibodies is clearly distinct from HLA-D antigens. Corresponding antigens seem to be expressed on placenta, since the lymphocytotoxic antibodies could be absorbed out with trophoblastic homogenates and one serum reacted with JAR cells (cultured choriocarcinoma line). An enhancing role of such antibodies in the growth of trophoblastic malignancies may be suggested. PMID- 6277038 TI - Bleb formation in granulosa cells of rat pre-ovulatory follicles in vivo and in vitro. AB - To determine if hormone-induced events leading to ovulation an granulosa cell luteinization might be associated with changes in the surface configuration of granulosa cells we have studied the morphology of granulosa cells from the preovulatory follicles both in vivo and in vitro. In vivo, granulosa cells in follicles from rats primed with estradiol and FSH developed bulbous protrusions termed blebs in response to injected hCG. The blebs were restricted to the adluminal granulosa cells which possess the least number of receptors for hCG. When granulosa cells from follicles of rats primed with estradiol and FSH were cultured in vitro, in the absence of serum, approximately 10% of the cells formed blebs. In the presence of 10% rat or fetal calf serum, nearly 90% of the cells formed blebs by 18 hr. Serum-induced bleb formation was prevented by 1 mM dibutyryl cycle-AMP plus 0.5 mM methyl isobutyl xanthine and by cytochalasin B (25 mug/ml), while 0.1 muM colchicine had no effect. Fibronectin at 25 mug/ml increased bleb formation three-fold over control values in serum-free medium. When hCG was included in serum containing medium, the majority of the cells remained smooth without any blebs. Thus, in contrast to its action in vivo, hCG inhibited the formation of blebs in vitro. When the cells incubated in the presence of dbcAMP plus methyl isobutyl xanthine in serum-containing medium, none of the cells formed blebs. One explanation for the seemingly opposite actions of hCG in vivo and in vitro is that hCG might act to alter the permeability of the pre-ovulatory follicles, and thereby allow the admission of serum. The admitted serum component(s) could then induce the formation of blebs on receptor-deficient adluminal cells that did not have elevated cAMP concentrations. The results suggest that fibronectin and/or other serum components, act to induce microfilament-dependent, cAMP-inhibited bleb formation on granulosa cells in vivo and in vitro. PMID- 6277039 TI - The distribution of cationized ferritin receptors on ciliated epithelial cells of rat trachea. AB - The interaction of tracheal cilia with the biphasic mucus layer covering the surface of the mammalian respiratory tract may be influenced by many cell surface coat components including those having an overall negative charge. In order to assess the distribution of ciliary anionic sites, cationized ferritin (CF) was used to label the surface of rat tracheal epithelium. If pieces of trachea were fixed with 3% glutaraldehyde and treated with CF at low (L) (0.08 mg/ml), medium (M) (0.32 mg/ml PBS), or high (H) (0.64 mg/ml PBS) concentrations, the label was distributed evenly over the entire external surface of the ciliary membrane at all concentrations. Unfixed tracheal tissue was also treated with L, M, and H CF for 1 or 5 min at 4 degrees C in order to minimize lateral redistribution of CF receptors. To ensure accessibility of the cell surface to CF the samples were agitated thoroughly during exposure. Exposure for 1 min to L, M, and H CF resulted in a light binding of ferritin particles on all portions of the ciliary membrane with occasional areas of multilayered binding distributed randomly on the ciliary shaft. When unfixed trachea was treated with CF for 5 min at 4 degrees C, CF binding was similar except heavier and more uniform. In no instance was there any preferential binding of CF to the ciliary tips at any of the concentrations used. Moreover, as indicated by the CF binding pattern at L concentrations, high density negative charges are present over almost the entire surface of the cilium. These results suggest that, unlike the ciliary membrane of other organs such as oviduct, negatively charged cell surface coat molecules are present on all areas of the ciliary membrane of rat tracheal epithelia. PMID- 6277040 TI - Effect of sodium intake on brain angiotensin-converting enzyme activity in spontaneously hypertensive rat. AB - The effect of sodium intake on angiotensin-converting enzyme activity in five areas of the brain (the cerebral cortex, midbrain, striatum, thalamus and hypothalamus) was studied in normotensive, spontaneously hypertensive and stroke prone spontaneously hypertensive rats. The enzyme activity was significantly higher in the hypothalamus than in other areas of the brain of spontaneously hypertensive rats. Sodium intake resulted in a significant rise of the enzyme activity in the midbrain of spontaneously hypertensive rats and also in the midbrain and the striatum of stroke-prone spontaneously hypertensive rats. In normotensive rats, however, there was no significant difference in the enzyme activity in any area of brain between the control and the salt-treated group. It is likely therefore that a high circulating sodium level increases angiotensin converting enzyme content of the brain in spontaneously hypertensive rats, and it is suggested that the increased converting-enzyme activity may play a role in development of hypertension induced by sodium loading. PMID- 6277041 TI - The effects of adriamycin in vitro and in vivo on hepatic microsomal drug metabolizing enzymes: role of microsomal lipid peroxidation. PMID- 6277042 TI - Protection of mice against lethal effects of sodium arsenite--a quantitative comparison of a number of chelating agents. PMID- 6277043 TI - Pardachirus marmoratus (Red Sea flatfish) secretion and its isolated toxic fraction pardaxin: the relationship between hemolysis and ATPase inhibition. PMID- 6277044 TI - Tissue distribution and elimination kinetics of polybrominated biphenyls (PBB) from rat tissue. AB - The concentration of polybrominated biphenyl (PBB) in serum in a large number of organs was determined in a population of rats for 36 weeks a single dose of PBB. Groups were killed at 6, 12, 24 and 36 weeks after exposure to PBB (1 mg/100g body wt, i.p.). Growth, weight gain and appearance of the rats and their internal organs were normal. Complex and varied relationships were found in tissue concentrations with time after PBB administration. Serum and fat had apparent first-order elimination kinetics with calculated half-times of 23.1 and 69.3 weeks, respectively. For five other tissues, apparent t 1/2s ranged from 9.0-63 weeks, while for four others, kinetics could not be determined from these 4 time points. It is likely that a substantial residue of PBB will still remain in the body of the rat at the end of its life span because of the persistence of PBB in lipid-rich tissues (adipose, adrenal, and brain). PMID- 6277045 TI - Pesticide-induced physiological alterations in certain tissues of a fish, Mystus vittatus. AB - Effect of sublethal concentrations (1/5, 1/10 and 1/15 fractions of 96-h TL50) of thiotox, dichlorvos and carbofuran on acid, alkaline and glucose-6-phosphatase of liver, kidney and gills of Mystus vittatus have been studied. Maximum (62.79%) inhibition of alkaline phosphatase in gills after thiotox (0.00013 mg/l) treatment have been observed. At 0.000045mg/l concentrations of thiotox, a marginal (7.6%) and significant (P less than 0.01) stimulation was observed in hepatic alkaline phosphatase activity. PMID- 6277046 TI - Enhancement of benzo(a)pyrene-induced sister chromatid exchanges as a consequence of glutathione depletion in vivo. AB - Pretreatment of Chinese hamsters with phorone (diisopropylidene acetone) decreased hepatic glutathione (GSH) content to about 20% of the control level after 2 hours. The GSH S-transferase activities were not affected. As a consequence of reduced detoxication by GSH the potency of 3,4-benzo(a)pyrene to induce sister chromatid exchanges in vivo in bone marrow cells was significantly enhanced. Chemically induced distinct alterations in metabolism of mutagens/carcinogens are proposed as experimental models for pharmacogenetic and toxigenetic studies. PMID- 6277047 TI - [Radiotherapy of adenomyosarcomas in children (author's transl)]. AB - A combined treatment with surgery, radiotherapy and chemotherapy is necessary for children with adenomyosarcomas. In order to reduce the side effects of the treatment, it seems reasonable to establish a therapy plan taking into consideration the risk of recurrences. Besides the classification of stages according to the N.W.T.S., the following factors should be considered: histologic findings, manifestations of the lymph nodes, number of tumor nodes, size of the adenomyosarcoma, and a rupture of the adenomyosarcoma. A postoperative polychemotherapy is sufficient for children of stage I presenting favorable histological findings. For the stages II and III, an irradiation of the tumor and the lymph outflow regions or the total abdomen with 24 Gy is recommendable in addition to chemotherapy. For stage IV, an irradiation of the lung up to 12 Gy in addition to chemotherapy has proved to be useful. A pre-operative radio- or chemotherapy is indicated especially in case of a great tumor volume and stage III and IV. PMID- 6277048 TI - Ultrasonic diagnosis of abdominal disease in Kenya. AB - This study presents the results of abdominal ultrasonic scanning in 108 patients attending a tropical referral hospital. Clinical diagnoses included hepatocellular carcinoma, metastatic liver disease, amoebic liver abscess, hydatid disease, obstructive jaundice, hepatosplenomegaly of uncertain aetiology and renal cysts and tumours. Because of its ability to distinguish solid from fluid-filled lesions, we found ultrasonic scanning the most useful initial investigation for the differentiation of hepatic masses. Ultrasonography is also ideal for the diagnosis of abdominal cysts and is extremely reliable in differentiating extrahepatic from intrahepatic obstructive jaundice. It is a non invasive procedure, quick and easily repeatable and has great potential in tropical medical practice. PMID- 6277049 TI - Dihydroorotate dehydrogenase, orotate phosphoribosyltransferase and orotidine-5' phosphate decarboxylase in Plasmodium falciparum. PMID- 6277050 TI - Multifocal demyelinative lesions of peripheral nerves in experimental chronic Chagas's disease. PMID- 6277051 TI - Myeloperoxidase activity in polymorphonuclear neutrophils in nutritional anaemia. PMID- 6277052 TI - Liver-fluke infection as an aetiological factor in bile-duct carcinoma of man. AB - This paper reviews, with particular reference to Opisthorchis viverrini, the evidence that opisthorchiasis and clonorchiasis are aetiological factors in the causation of human bile-duct cancer, especially cholangiocarcinoma, and considers the other aetiological factors which may be operating. The epidemiology, pathology in man and experimental animals, and histogenesis of cholangiocarcinoma associated wtih liver-fluke infection are also considered. PMID- 6277053 TI - Significance of mitochondrial enhancement in restoring hepatic energy charge after revascularization of isolated ischemic liver. AB - The critical point of reversibility in hepatic ischemia was investigated. Immediately after the liver was isolated, the hepatic energy charge decreased rapidly from the control level of 0.85 to 0.28 and remained at a constant value during 30-, 60-, and 90-min ischemic periods. After revascularization of the liver, the rapidity toward the normalization of the energy charge and the mitochondrial phosphorylative activity were different according to the ischemic time. The livers after 30- and 60-min exclusion showed a normalization of the energy charge and the enhancement of mitochondrial phosphorylative activity, while the livers after 90-min exclusion showed only a slight increase of energy charge, which remained far from the normal value, and the enhancement of mitochondrial phosphorylative activity was not observed. From these results, it is suggestive that a rapid enhancement in phosphorylative activity of the liver mitochondria after revascularization plays a decisive role on the restoration of the viability of isolated ischemic liver. PMID- 6277054 TI - Reversal of diabetes by the isotransplantation of nicotinamide-streptozotocin induced islet adenoma in rats. AB - The effect of syngeneic transplantation of islet tumors induced by streptozotocin nicotinamide treatment into Fisher inbred diabetic rats was studied. Transplantation of 10 mg of tumor tissue under the kidney capsule of severely diabetic animals resulted in complete amelioration in 9 of 14 animals. Partial improvement was noted in three animals, whereas only one animal showed no change in diabetic condition within 4 months. Improvement in diabetes was accompanied by gain in body weight, decrease in blood sugar level and urinary glucose excretion, and normalization of glucose tolerance. After removal of the kidney containing the tumor grafts, the animals reverted to a severe diabetic condition, suggesting that the tumor graft supplied the insulin needs of the diabetic animals. The graft tissue exhibited in vitro insulin synthesis and secretion in response to 16.7 mM glucose. PMID- 6277055 TI - B cell, helper T cell, and suppressor T cell abnormalities contribute to disordered immunoglobulin synthesis in patients following bone marrow transplantation. AB - The pathogenic mechanisms responsible for the hypogammaglobulinemia that occurs in patients who have undergone bone marrow transplantation were studied using peripheral blood lymphocytes in an in vitro immunoglobulin biosynthesis assay. None of the nine marrow recipients between 2 and 15 months after transplantation produced normal amounts of IgG, IgA, or IgM in response to pokeweed mitogen (a T cell-dependent activator). However, three of these same patients responded to Epstein-Barr virus (a helper T cell- independent activator), suggesting that these three possessed responsive B cells. Cocultures of the marrow recipients' lymphocytes with lymphocytes from their respective donors or other normal subjects documented excessive suppressor cell activity in five cases. These suppressor cells frequently had profound activity (greater than 80% suppression of the immunoglobulin synthesis of cocultured normal cells), were predominantly radioresistant, and were T cells in the cases where analyzed. Helper T cell activity for immunoglobulin synthesis was not demonstrable in seven of the nine cases. In addition, four of the patients apparently possessed defective B cells. Although most patients had combined defects of several lymphocyte subpopulations, it appeared that either an isolated helper T cell or isolated B cell deficiency was sufficient to result in altered Ig synthesis. In this regard, two patients had demonstrable helper T cell defects with partially responsive B cells and no excessive suppressor activity. Interestingly, these two patients were experiencing a recurrence of their hypogammaglobulinemia following a period of normalized immunoglobulin levels. Thus, defects within all lymphocytic elements involved in the response and regulation of immunoglobulin synthesis were identified and may contribute to the humoral immunodeficiency which follows marrow transplantation. PMID- 6277056 TI - Heritable differences among gp70-like molecules on C3H ultraviolet light-induced fibrosarcomas. PMID- 6277057 TI - Obtention of 7 ascites tumor lines expressing H-2s and not H-2k from a C3H specific polyoma-induced solid tumor. PMID- 6277058 TI - H-2D control of radiation leukemia virus induced neoplasia: evidence for interaction of viral and H-2 genomic information. PMID- 6277059 TI - Characterization of alloantigen-activated human lymphocytes cytotoxic for autologous EB virus-transformed lymphoblastoid cell lines. PMID- 6277060 TI - The regulation of the human antitumor immune response to organ-specific neoantigens. PMID- 6277061 TI - Anti-BROC-A1 (anti-H-2s) antibodies cross-react with HLA-A11. PMID- 6277062 TI - [Independence of the insulin effect from the guanosine-3',5'-monophosphate level in muscle tissue]. AB - The role of cyclic GMP in the insulin effect was investigated using isolated frog sartorii. A study was made of the effect of exogenous cyclic GMP, dibutyryl cyclic GMP, 8-bromo-cyclic GMP on xylose transport, glycogen synthesis and muscle respiration. Only dibutyryl cyclic GMP (1.10(-6) - 10(-4) M) alone was observed to have a stimulating effect on glycogen synthesis and respiration. The xylose transport was but slightly accelerated only following a 20 hours incubation of muscles in the cyclic GMP solution. Cyclic GMP was shown to penetrate the muscle fibres. The cyclic GMP content in muscles was equal to 22.7 +/- 2.0 pM per gram of wet weight. Insulin exerted no effect on cyclic GMP concentration in muscles. The data obtained do not allow to conclude that cyclic GMP may serve as a mediator in realization of the insulin effect on membrane and intracellular processes. PMID- 6277063 TI - [Individual renal function determinated by 99mTechnetium-dimercaptosuccinic acid and 131I-hippuran]. PMID- 6277064 TI - [Antibodies to hepatitis A virus in blood donors. Relationship to age distribution of blood donors]. PMID- 6277065 TI - Invasive lobular carcinoma of the breast. PMID- 6277067 TI - Wilms tumor with consumption coagulopathy. PMID- 6277066 TI - Preoperative chemotherapy for Wilms tumor. AB - Preoperative chemotherapy was administered to 19 children with Wilms tumor judged clinically to be unresectable at M. D. Anderson Hospital between January 1, 1962, and September 1, 1980. After 2 to 4 doses of vincristine, marked reduction in tumor size occurred in 16 patients. After chemotherapy 16 tumors could be resected completely, another required irradiation to reduce the tumor, and only 2 tumors could not be excised. Pathologically the most dramatic changes occurred in the undifferentiated interstitial stroma, followed next by changes in the nodular blastema. Differentiated elements were apparently not affected. No serious complications were attributed to the preoperative drug treatment. This experience suggests that in selected instances preoperative chemotherapy can affectively facilitate the therapy of Wilms tumor. PMID- 6277068 TI - Compensatory renal growth and function in postnephrectomized patients with Wilms tumor. AB - The objective of this study was to determine whether or not renal growth and function were adversely affected in the remaining kidneys of patients who had undergone nephrectomy for Wilms tumor. These patients received chemotherapy and some radiotherapy (tumoricidal agents which might affect the remaining kidney). Renal growth was compared between the treatment groups and normal renal growth. Hypertrophy did occur and did not appear to be affected by subsequent treatment. Renal function was minimally altered in all treatment groups irrespective of the type of treatment. PMID- 6277069 TI - Adult Wilms tumor. Treatment with surgery, radiotherapy, and chemotherapy. PMID- 6277070 TI - Cystic mesoblastic nephroma. AB - Mesoblastic nephroma is the most common solid renal neoplasm in the first few months of life. It has been recognized only recently as a distinct entity, having been classified previously as Wilms tumor. There are relatively few ultrasonic descriptions of this entity in the literature. We present an additional case featuring a large peripheral cystic component with a solid central core demonstrated on ultrasonograhy. This feature is not specific for mesoblastic nephroma and occurs in cystic Wilms tumor. PMID- 6277071 TI - Effects of PGI2 and di-butyryl cyclic-AMP on platelets exposed to shear stress. PMID- 6277072 TI - Effects of drugs on adhesion of human platelets. PMID- 6277073 TI - Plasma protein adsorption and desorption rates on quartz: approach to multi component systems. PMID- 6277074 TI - High sodium dialysate with addition of chloride, acetate and bicarbonate. PMID- 6277075 TI - [Fibrohistiocytosis of the stomach]. PMID- 6277076 TI - The pancreas--2: diabetes mellitus. PMID- 6277077 TI - Poison caused by ingestion of chokecherry leaves in a dog. PMID- 6277078 TI - Ketoconazole treatment of osseous blastomycosis in a dog. PMID- 6277079 TI - Cushing syndrome in dogs: treatment with bromocriptine and cyproheptadine. PMID- 6277080 TI - [Detection of PI-3 viruses, BVD-MD viruses and parvoviruses in bovine fetuses after experimental intrauterine infections]. AB - Growing organ cultures prepared from foetuses experimentally infected in utero by the viruses PI-3 and BVD-MD or bovine parvovirus proved to be suitable in tests with the re-isolation of these viruses 7, 14, 21, 35, 42 and 70 days after infection. The attempts at demonstrating them by the traditional method of the inoculation of the primary cell culture of foetal kidneys were successful only at the re-isolation of the PI-3 virus seven days after infection. The PI-3 virus without cytopathic effect was demonstrated in the primary cell culture prepared from the spleen, lungs, kidneys and testes of foetuses delivered seven days from infection. The cytopathic effect manifested itself after cell transplantation in the first passage when further multiplication of the virus occurred. PMID- 6277081 TI - [Multiplication of the IBR, PI-3 and BVD-MD viruses in cell and organ cultures of bovine fetuses after experimental intrauterine infections]. AB - The intrauterine infection of four- to nine-month-old bovine foetuses with the PI 3, BVD-MD viruses, performed 7 to 72 days prior to their delivery, did not exert any significant influence upon the susceptibility of primary cell cultures from foetal organs and tissues to further viral infection in vitro. The BVD-MD and IBR viruses multiplied in the primary cell cultures from the organs of a foetus infected with the PI-3 virus seven days before delivery even in the presence of endogenous PI-3 virus. Persisting infection with the PI-3 virus also failed to influence the susceptibility of foetal organ cultures to infection with the IBR and PI-3 viruses in vitro. The IBR virus and endogenous PI-3 virus multiplied simultaneously to high titres in the organ cultures of thymus and lungs whereas in the organ cultures of kidneys, spleen and testes the multiplication of endogenous PI-3 virus was suppressed. PMID- 6277082 TI - Plant induced calcinosis: a review. PMID- 6277083 TI - In vitro and in vivo investigations on reticuloendotheliosis virus infection. AB - Experimental transmission of reticuloendotheliosis (RE) in 1-day-old SPF chickens was achieved by intraperitoneal inoculation of cell-associated and cell-free RE virus (REV). REV-inoculated chickens exhibited an immune response throughout the entire observation period. Viremia persisted for 5--6 months in chickens hatched from REV-inoculated SPF embryonated eggs. The natural occurrence of RE was detected in turkey flocks of farms Bc. PMID- 6277084 TI - Comparative results of respiratory virus isolation attempts in chorioallantoic membrane fragments and embryonated chicken eggs. AB - Attempts performed with 39 samples of nasopharyngeal secretion resulted in the isolation of 21 (54%) hemagglutinating viral agents in chorioallantoic membrane (CAM) fragments, while a single strain (2.5%) was isolated in the embryonated egg. The time interval required for virus isolation in CAM fragments ranged from 4 to 20-24 days and was longer then 10 days in the embryonated egg. Maintenance of CAM fragments in roller tubes leads to a more abundant virus multiplication and, implicitly, to a reduction of the time interval necessary for virus isolation. PMID- 6277085 TI - [Tumor specific surface antigens and immunological reaction of Rous sarcoma (review) (author's transl)]. PMID- 6277086 TI - [Basic research trends in the prevention of traumatic shock using pharmacological agents (2)]. PMID- 6277087 TI - [Case of anhidrotic ectodermal dysplasia]. PMID- 6277088 TI - [Kidney tumors in biopsy material 1958-1979]. PMID- 6277089 TI - [Case of cystosarcoma phyllodes in mesenchymoma malignum vertens]. PMID- 6277090 TI - Treatment of patients with small-cell lung cancer at the Memorial Sloan-Kettering Cancer Center, 1974-1979. PMID- 6277091 TI - Minute liver cancer and concomitant esophageal varices: detection and successful surgical treatment. PMID- 6277092 TI - [Distribution of transferable trimethoprim and gentamicin resistance plasmids in Enterobacteriaceae]. AB - Following the employment of trimethoprim/sulfonamid and gentamicin in the general clinical praxis transferable plasmids with respective resistance function were identified at first in enteric bacteria from sewage before they could be detected in patient strains. Whereas the trimethoprim resistance plasmids represent different incompatibility groups (C, FII, I5, K, M, N, S, U) the gentamicin resistance plasmids are very similar IncM members of 62 MD in size. Therefore a clonal distribution of a particular gentamicin resistance plasmid has to be taken into consideration. PMID- 6277093 TI - [The influence of oxygen and bicarbonate administration to the parturient on maternal and fetal blood gases and acid base balance (author's transl)]. AB - The effects of oxygen and bicarbonate administration to parturients were studied by analysis of maternal and fetal blood gas parameters. Oxygen inhalation resulted in a significant increase of the fetal pO2 and appears to be a practicable mean of intrauterine reanimation of hypoxic fetuses. Intravenous bicarbonate was unable to raise the fetal pH. Simultaneous bicarbonate and oxygen administration had no desirable effects on the fetal acid-base-balance. PMID- 6277094 TI - Radiosynoviorthesis in haemophilic joint disease. AB - The effects of radiosynoviorthesis on chronic haemophilic arthropathic joints were studied in six patients with severe haemophilia by a follow up study which spanned two and a half years. On clinical grounds the treatment was successful because pain and bleeding frequency diminished, although radiographic examination showed further deterioration of the treated joint. Chromosome damage was not detected. We conclude that radiosynoviorthesis is apparently changing the bleeding pattern of articular tissues without arresting the destruction and deformation of joints. PMID- 6277095 TI - [Histological, electron microscopical and X-ray microanalytical findings in malakoplakia of the kidney (author's transl)]. AB - Malakoplakia which was found in the kidney if a man aged 55 years is described. This rare granulomatous lesion was extirpated together with a renal cyst. So called Hansemann-cells with inclusion bodies and ring structures were found by light and electronmicroscopic examinations. These concentrically arranged ring like structures were not typical Michaelis-Gutmann-bodies, and they consisted of microfibrils and small granula. Using X-ray microprobe analysis could be detected calcium. These findings were considered to be an early stage of malakoplakia. PMID- 6277096 TI - [On the biological action of transition metal complexes. 3. About the antiviral activity of cis-dichloro diammine platinum (II) (author's transl)]. AB - The coordination compound cis-dichlorodiammineplatinum(II) (cis-DDP) was shown by Rosenberg et al. (17) to exhibit antitumour activity. Several authors have indicated limited virustatic properties of cis-DDP against bacterial, oncogenic, avipox and paramyxo viruses. In our investigations, cis-DDP significantly showed an antiviral action in vitro against enveloped DNA and RNA viruses, such as vaccinia, pseudorabies, herpes simplex type 1, Newcastle disease, influenza A/fowl plague, influenza A/Victoria 3/75, influenza A/Jena 48/78, influenza B/Johannesburg and vesicular stomatitis viruses. Out of the group of nonenveloped viruses, adenovirus type 4 and 5 were inhibited, whereas no inhibition against naked cardiovirus Mengo could be estimated. The antiviral action was proved against extracellular virus by dialysis experiments with vaccinia virus and also during the replication cycles of enveloped viruses. In trials with cell-free viruses the plaque reduction of all sensitive viruses mentioned above amounted to 100 per cent in comparison to the untreated controls caused by virus inactivation with loss of infectivity in contact with several concentrations of cis-DDP. On the other hand, the addition of the compound for one hour only immediately after infection or up to 8 hrs later produced a complete depression of further multiplication of vaccinia virus. Likewise, the replication of influenza virus A/FPV or VSV was inhibited whereas the multiplication of adenoviruses was not influenced in a comparable manner. PMID- 6277097 TI - [On the biological action of transition metal complexes. 4. The antiviral activity of metallocene dichlorides of titanium and molybdenum (author's transl)]. AB - In view of the fact that bis cyclopentadienyl metal dihalides are known to be anti-tumour drugs, we have investigated the antiviral activity of this type of coordination compounds. Bis cyclopentadienyl titanium dichloride (a) has shown significant antiviral efficiency in vitro against representatives of a nuber of enveloped DNA and RNA viruses. Inhibition of orthopoxvirus (vaccinia), herpes virus (pseudorabies), orthomyxoviruses (influenza A/fowl plague [FPV], influenza A/Victoria 3/75, influenza A/jena 48/78 and influenza B/Johannesburg), paramyxovirus (Newcastle disease [NDV]) and rhabdovirus (vesicular stomatitis [VSV]) was observed after direct contact with the compound under loss of infectivity up to 100%. Regarding the group of unenveloped viruses only adenovirus type 4 became influenced but not type 5. No antiviral activity could be found against the cardiovirus Mengo. The compound bis cyclopentadienyl molybdenum dichloride failed to show an antiviral action versus vaccinia, influenza A/FPV and influenza viruses B/Johannesburg. Application of the inhibitor (a) during the replication of vaccinia and influenza viruses A/FPV in cell cultures produced an additional effect of inhibition of virus multiplication. On the other hand, adenovirus type 4 and VSV replication was not affected by titanocene dichloride. PMID- 6277099 TI - High and prolonged incidence of poliovirus strains during 1979 in North Bohemia. Isolations from unusual materials. AB - During March 1979 - February 1980, 53 strains of polioviruses types 2 and 3 were isolated from human specimens and from sewage samples, 52.8% of them on secondary monkey kidney cultures, 47.2% on diploid fibroblasts of human embryonic lung. Type 3 grew more often on monkey cell cultures, type 2 on human ones. Both types of poliovirus circulated in children and adult population during the whole year. Polioviruses were isolated from cerebrospinal fluids of 2 patients without any symptom of paralytic poliomyelitis: an intermediate type 3 in the course of a serologically proved tick-borne encephalitis, and a type 2, different in rct marker from the vaccinal variant, in the course of a serologically proved parotitis with meningoencephalitis and pancreatitis. PMID- 6277098 TI - [Investigations of the enzyme spectrum of Erysipelothrix rhusiopathiae (author's transl)]. AB - The enzyme spectrum of non proliferating cells of Erysipelothrix rhusiopathiae was investigated by means of different low molecular synthetic substrates. Activities of aminopeptidases were found directed against compounds of L-alanine, L-arginine, L-aspartic acid, glycine, L-isoleucine, L-leucine, L-lysine, L methionine, L-phenylalanine, L-proline, L-tryptophane, and L-tyrosine, but not against compounds of l-cystine, L-glutaminic acid, L-histidine, L-hydroxyproline, and L-valine (Table 1). The pH optimum of the investigated aminopeptidases ranges from neutral to alkaline reaction (Table 2). Trypsin, chymotrypsin, or chymotrypsin-like proteases were not detected. E. rhusiopathiae possess esterase activity splitting esters of lower carboxylic acids, i. e. acetic acid, propionic acid, butyric acid, caproic acid, and caprylic acid, but no lipase activity. Under the provoked glycosidases only alpha- and beta-D-galactosidase and glucosaminidase were positive. Weak activities of phosphatases and arylsulfatase were found also (Table 3). PMID- 6277101 TI - [In vitro susceptibility of Listeria monocytogenes with special reference to newer beta lactam antibiotics (author's transl)]. AB - Minimal inhibitory (MIC) and minimal lethal concentrations (MLC) of ampicillin, mezlocillin, piperacillin, cephalothin, cefamandole cefoxitin, cefuroxime, cefotaxime, gentamicin, and doxycycline against 44 strain of Listeria monocytogenes from clinical specimens were determined by broth dilution tests. With regard to the MIC90 and MLC90 values (concentrations which inhibited or killed 90 percent of the strains) ampicillin showed the greatest anti-listeric activity with 0,48 and 20,8 microgram/ml. The corresponding results for mezlocillin were 8,0 and 54,4 microgram/ml, for piperacillin 5,2 and 128 microgram/ml, resp. Among the cephalosporins, cephalothin showed the best activity (MIC90, 5,8 microgram/ml, MLC90, 76,8 microgram/ml). The MIC90 values of cefamandole and cefoxitin were 7,8 and 89,6 microgram/ml, resp. and the MLC90 was more than 128 microgram/ml while there was no clinically relevant activity of cefuroxime and cefotaxime (MIC90 more than 128 microgram/ml). The MIC90 and MLC90 results of gentamicin were 12 and 15,2 microgram/ml, resp, and those of doxycycline 8 and 25,6 microgram/ml, resp. There were only slight differences between gentamicin MIC and MLC values. Henceforward the drug of choice for the treatment of generalized listeric infections should be ampicillin eventually in combination with gentamicin. PMID- 6277100 TI - Quantification of amebae specific antibodies as "Multiple of normal activity (MONA)" with a standardized enzyme immunoassay (EIA). AB - Sera of 853 returnees from tropical countries, of 24 cases with amebic liver abscess and of 172 nonexposed German individuals were tested for antibodies to E. histolytica in the enzyme immunoassay (EIA). Antibody results were expressed as "multiple of normal activity (MONA)". The qualification of the EIA as a routine screening procedure for amebae specific antibodies was investigated and compared to that of the complement fixation test. Based on the symmetric frequency distribution of results from the 172 non-exposed controls the upper one sided limits for 95% (less than 2.8 MONA) and 99% (less than 4.2 MONA) specificity were determined. Routine results below the 95% specificity limit were considered "inconspicuous", such between both limits "borderline" and all MONA values exceeding the 99% specificity limit "conspicuous". The intention was to thereby secure a high degree of sensitivity for amebae antibody in the test. Cases with clinically confirmed liver abscess revealed a one sided lower 95% sensitivity limit (greater than or equal to 1.2 MONA), far above the onset of the defined sensitivity thresholds for conspicuous MONA values. PMID- 6277102 TI - [Differentiation of pathogenic clostridia from material collected in the field and of reference strains by means of biostatistical analysis of the quantitatively recorded metabolic fatty acid patterns of the agents (author's transl)]. AB - 34 reference and 9 exotic field strains of clostridia were analyzed by means of gas liquid chromatography (GLC) on their patterns of metabolic short chain fatty acids. To increase the number of criteria for the differentiation specimens were grown on pepton yeast medium with addition of 13 different carbohydrates and alcohols. Data of sequence and mean square space of GLC-peaks were stored in a data bank. With the SPSS discriminance analytical system the field strains were compared with the reference strains in order to classify field to reference strains. At the same time differences between reference strains of different origin were determined. C. novyi, C. histolyticum., C. sordellii, C. difficile, C. subterminale and C. sporogenes could be separated clearly from the C. chauvoei , C. septicum- and the perfringens-group. Between different strains of C. chauvoei provided by several laboratories significant differences could be shown which make original classification doubtful. Similarities between the C. chauvoei and the C. septicum-group do exist. Formerly C. perfringens type F is the sole type of C. perfringens which can be separated from the C. perfringens-group. There are similarities in fatty acid patterns between the C. chauvoei-, C. septicum-, and C. perfringens-group. Strains isolated from field cases of gas gangrene in Peru, Madagascar and Germany (Bavaria) were classified as completely different from the reference strains for Peru, partly similar to C. chauvoei for Madagascar, similar to C. chauvoei and C. septicum for Bavaria. PMID- 6277103 TI - [Studies on the induction by extracts from airborne particulates of benzo(a)pyrene hydroxylase and the binding of benzo(a)pyrene to DNA in the rat lung in vivo after pertracheal administration (author's transl)]. AB - A method is described for the administration of suspended material into the rat lung. The rat is narcotized by CO2/air, the trachea is punctured and the suspension is sucked into the lung by the inhaled air (pertracheal administration, p. t. a.). BaP administered by this method is eliminated from the lung rapidly, while metabolites persist for a longer period. The radioactivity detected in the blood can be attributed predominantly to metabolites, a small fraction is unmetabolized benzo(a)pyrene. The activity of benzo(a)pyrene hydroxylase is induced in the lung after p. t. a. of extracts from airborne particulate matter. The activity increases depending on the dose until a plateau of activity is reached at high doses. Pertracheally administered BaP is bound to DNA of the lung. Pretreatment with extracts p.t.a. enhances the binding. After enzymatic digestion of DNA BaP-nucleoside adducts are detected by HPLC. According to data published in the literature to adducts are supposed to be derived from BaP-phenols and BaP-diolepoxides. It is concluded tht the components of airborne particulates can modify the activity of metabolizing enzymes and increase the rate of formation of adducts in the lung in vivo. PMID- 6277104 TI - [Vitamin D3 and vitamin D3 metabolite-like activity in Trisetum flavescens]. PMID- 6277106 TI - Nephroblastoma in an ovine foetus. PMID- 6277105 TI - [Interaction of gonadal receptors from rat testes with human chorionic gonadotropin: experimental implications of the dilution effect]. PMID- 6277107 TI - Light microscopy and ultrahistology of intestinal changes in pigs infected with epizootic diarrhoea virus (EVD): comparison with transmissible gastroenteritis (TGE) virus and porcine rotavirus infections. PMID- 6277108 TI - [Production of anti-enterotoxin antiserum to Clostridium perfringens type A toxin]. PMID- 6277109 TI - [Polyacrylamide gel disc electrophoretic study of laboratory and freshly isolated strains of Bordetella pertussis and Bordetella parapertussis]. AB - Differences in the antigenic complexes of laboratory and newly isolated B. pertussis and B. parapertussis strains have established by means of disc electrophoresis in polyacrylamide gel. B. pertussis strains have been found to contain 2 or 3 antigens more. Laboratory and newly isolated B. pertussis strains have been found to differ in antigenic complexes, the antigens in these complexes having different electrophoretic mobility. The antigens detected in newly isolated strains have greater electrophoretic mobility in comparison with that of the antigens detected in laboratory strains. No such differences have been revealed in laboratory and newly isolated B. parapertussis strains. Disc electrophoresis in polyacrylamide gel allows to detect 2 or 3 more antigens than immunoelectrophoresis in agar gel. PMID- 6277110 TI - [Non-selection mechanism of antibody-induced stable antigenic variability of microorganisms. I. Patterns in the antigenic variability of Bordetella pertussis]. PMID- 6277111 TI - [Cultivation of Bordetella pertussis in liquid semisynthetic nutrient media with ion-exchange resins]. AB - The possibility of optimizing the processes of B. pertussis cultivation carried out in liquid semisynthetic culture media with strong-base anion-exchange resins is shown. The use of strong-base ionites enhances the reproducibility of cultivation processes, increases the activity of microbial populations and the concentration of microorganisms. B. pertussis cultures grown in the media with anion-exchange resins are characterized by pronounced biological activity. PMID- 6277112 TI - [Rotavirus gastroenteritis]. PMID- 6277113 TI - Effect of sympathomimetic substances and analogues on tail absorption of frog larva. AB - Similarly to the effect of T3, the tail absorption of frog larvae (Rana arvalis) was stimulated in vitro by the sympathomimetic agents adrenaline, isoproterenol, ephedrine and the adrenaline analogue MMT. The effect of T3 administration was not inhibited by adrenaline, isoproterenol and MMT, but it was by ephedrine and T3 pretreatment. The adrenaline and isoproterenol effect manifested itself after early administration, but not after late and while ephedrine and MMT were effective at long-term administration, even it administered at a later time. Our experiments draw attention to the metamorphotic effect of substances capable of binding to beta-receptors. Their effect may be explained partly by the increase in the cAMP level, partly by a relationship existing between the beta-receptor and the T3 receptor. PMID- 6277114 TI - Morphogenetic effect of cAMP during the metamorphosis of the frog larva. AB - In an in vitro system, the tail absorption of the frog larva was significantly stimulated by cAMP more intensely than by T3. Theophylline, an inhibitor of cPDE, had no effect and failed to enhance, even decreased the effect of T3. It may be supposed that T3 does not act through a cAMP mechanism, but the tail absorption can be influenced by cAMP in a direct way. PMID- 6277115 TI - Influence of linseed oil diet on the pattern of serum phospholipids in man. AB - We gave 11 volunteers 30 ml linseed oil daily for 4 weeks. This diet increased the amount of linolenic, eicosapentaenoic and docosahexaenoic acid in the serum phospholipids within 1 week, whereas the amount of dihomo-gamma-linolenic acid was decreased. Both effects were statistically significant. The results suggest that eicosapentaenoic acid can be synthesized from alpha-linolenic acid in human organism. PMID- 6277116 TI - Properties of an acetylcholine-induced conductance in the rabbit atrial myocardium. AB - Using a conventional microelectrode technique, action potentials (A.P.) recorded from the isolated left atrial trabeculae of the rabbit were analyzed. The membrane current during A.P. was reconstructed. In spite of an extracellular Ca2+ deficiency and the application of verapamil, acetylcholine (ACh) reduced the A.P. duration by inducing an outward current IACh. This current was blocked by atropine (10-6 M). A Nernst-plot of the reversal potential at different K+ concentrations showed a slope of 58.5 mV for a 10-fold change in concentration. After pacing pauses longer than 10 s an inward going (anomalous) rectification (A.R.) for IACh occurred. Increasing the duration of the pacing pauses the rectification was more accentuated. During a constant pacing the A.R. for IACh disappeared. ACh did not modify the A.R. The maximum slope conductance for IACh was dependent on the extracellular concentration of ACh (0.035 mS x cm-2 at 20 microM ACh, 0.012 mS x cm-2 at 0.2 microM ACh). The experimental results are discussed, using the model of an ACh-induced potassium channel. The channel should be related to the muscarinic receptor of the atrial myocardium. PMID- 6277117 TI - Erythrocyte alterations in hemoglobin H disease. AB - This study on erythrocytes in hemoglobin H (Hb H) disease reveals that unstable Hb H is bound to membranes to a greater extent, especially when it forms methemoglobin or is precipitated as inclusion body. The methemoglobin content of these erythrocytes is elevated in spite of a higher activity of NADH methemoglobin reductase. The ATPase activity is doubled, and the ATP is presumably used for phosphorylation of membrane proteins, which leads to cross linking of membrane proteins. This assumption could be supported by the observed decrease in non-electrolyte permeability, by increased binding of hemoglobin to the membrane and by polymerisation of membrane proteins detected by SDS polyacrylamide gel electrophoresis. By means of electron microscopy, it could also be shown that the inclusion bodies are bound to the inner surface of membrane and cause its protrusion. This linkage might be responsible for the observed inhibition of the lateral movement of intramembrane particles. PMID- 6277118 TI - Separation by counter-current distribution of rat and chicken erythrocytes of different age, and its application to the assay of enzyme activities. AB - The separation of cells with different ages from erythrocyte populations of adult rats and young or adult chickens have been achieved by counter-current distribution (CCD). A thin-layer CCD apparatus has been employed. Erythrocytes from blood samples taken at different times after 59Fe i.p. injection were separated by CCD. By compilation in a "composite curve" of the hemoglobin and radioactivity CCD profiles obtained for each erythrocyte population, the distribution of cells according to age can be inferred. Young erythrocytes of rats are located at the right part of the CCD curves, while older cells are distributed towards the left. An opposite distribution has been found for erythrocytes from adult or young chickens. As a first attempt for the application of the CCD procedure to the assay of enzyme activities, it was found a decrease in phytase activity as the age of chicken erythrocytes increases and an increase in phosphoglycerate kinase and phosphofructokinase as the age of rat erythrocytes increases. PMID- 6277119 TI - Effects of dibutyryl-cANP on glycolysis in washed human erythrocytes. AB - The experiments were carried out with washed human erythrocytes in order to study the effects of dibutyryl-cAMP (DB-cAMP) on glycolysis. 5 mM DB-cAMP significantly increases glucose consumption and lactate production in incubated erythrocytes. The cross-over plot of glycolytic intermediates shows that increased glycolysis is probably the result of activation of phosphofructokinase by DB-cAMP. Under the same condition DB-cAMP significantly protects the 2,3-diphosphoglycerate level in incubated erythrocytes. PMID- 6277120 TI - Hepatic effects of glucose and insulin independent of cyclic nucleotide changes. AB - To determine if cGMP might function as a second messenger for insulin, an in situ liver perfusion system was established in which hepatic effects of insulin could be correlated with changes in cyclic nucleotides. Several combinations of insulin (10 mU/ml) and glucose (50 mg/ml) were infused (0.1 ml/min) for 30 min into fasted normal and diabetic rats with removal of a similar volume of blood. Samples of livers were removed at the beginning and end and at various times during the perfusion. In normal animals perfused with buffer alone, hepatic glycogen content fell. When glucose (with or without added insulin) was added to the perfusate, glycogen levels rose. With buffer alone, there was no change in the independent (I) form of glycogen synthase at 10 min but a modest increase at 30 min. With insulin and/or glucose, there as a large increase in the I-form of the enzyme at 10 min and a further rise at 30 min. Neither cGMP nor cAMP changed even though tissue samples were obtained at multiple times throughout the perfusion. Cyclic nucleotides were also measured in liver slices exposed to insulin (1 mU/ml) after 30 min of pre-incubation for stabilization. Although significant increases in cGMP were noted in the tissue exposed to insulin, similar significant rises also occurred in appropriately paired control slices. When glucagon was used in both the in situ perfusion and the paired liver slice systems, the expected rapid and large increases in cAMP levels occurred attesting to the validity of both approaches in evaluating hepatic cyclic nucleotide responses. These results plus the paucity of convincing data in the literature strongly suggest that cGMP can no longer be considered a candidate for the putative second messenger of insulin. PMID- 6277121 TI - Study of pituitary secretion in relation to retinopathy in patients with juvenile diabetes mellitus. AB - Fifteen juvenile diabetic patients with normal eye fundus, 6 with non proliferative retinopathy, 5 with proliferative retinopathy and 5 healthy control subjects were studied in order to investigate pituitary function in relation to diabetic retinopathy. ACTH values at 08(00) and 18(00), hPRL and TSh secretion in response to 200 microgram TRH i.v., and GH secretion in response to 500 mg oral L dopa were evaluated. In all diabetic subjects, 08(00) ACTH levels were lower than in controls. Basal hPRL, TSH and GH values of the diabetics did not differ from those of the controls. No significant differences were found in hPRL levels in response to TRH, whereas significantly lower TSH responses were found in the diabetics. L-dopa caused a significantly different response of GH in the diabetic subjects compared to controls. There is thus a considerable derangement in pituitary hormone secretion in juvenile diabetics mellitus. PMID- 6277123 TI - Cyclic nucleotides in ascidian embryonic development. I. Cyclic nucleotides determination. PMID- 6277122 TI - Effects of two cholecystokinin variants, CCK-39 and CCK-8, on basal and stimulated insulin secretion. AB - The effects of two different cholecystokinin variants, CCK-39 and the carboxyl terminal octa-peptide CCK-8, on basal and stimulated insulin secretion were studied in the mouse. It was found that both peptides had a dose-dependent stimulating action on insulin secretion with a maximal response of similar magnitude at a dose level of about 5 nmol/kg body weight. This dose induced an increase of plasma concentrations of immunoreactive insulin of about 100 microU/ml. The calculated half-maximal dose was 2.12 nmol/kg for CCK-39 and 3.18 nmol/kg for CCK-8. The insulin-secretory response to CCK-39 and CCK-8 in the absence of other secretagogues was partially abolished by pretreatment with the cholinergic blocker methylatropine as well as with the beta-adrenoceptor blocker L-propranolol. Thus, this great insulin-secretory response to the two peptides seemed to be dependent on intact muscarinic and beta-adrenergic receptors. CCK-39 or CCK-8 administered in a threshold dose prior to half-maximal doses of D glucose, the cholinergic agonist carbachol, or the beta-adrenergic agonist L isopropylnoradrenaline (L-IPNA), respectively, displayed different influences on insulin release. CCK-39 potentiated glucose- as well as carbachol-induced insulin secretion, whereas it did not influence L-IPNA-induced insulin release. An equimolar dose of CCK-8, on the contrary, had no apparent effect on either glucose-, carbachol-, or L-IPNA-induced insulin release. This observation indicates that stimulated insulin release is influenced by amino acid sequences other than those of the C-terminal octapeptide in CCK-39 and that the response of the stimulated insulin-secreting cells to CCK-39 is dependent on the nature of the secretagogue. PMID- 6277124 TI - Cyclic nucleotides in the amphibian discoglossus pictus during embryonic development. I. Cyclic nucleotides determination. PMID- 6277125 TI - Alterations in hepatic nuclear binding of triiodothyronine in experimental diabetes mellitus in rats. PMID- 6277126 TI - Effect of exogenous beta-endorphin on anterior pituitary hormone secretion in man. AB - The effect of large amounts of synthesized human beta-endorphin (beta-EP) administered intrathecally on pituitary-adrenocortical function was investigated by determining the plasma levels of ACTH, cortisol, growth hormone and prolactin in 8 patients with pain caused by severe disseminated cancer. They were divided into 2 groups, an Ep group of 8 patients and a control group of 5 of the same 8 patients. There were no significant effects of beta-Ep on plasma ACTH, cortisol and growth hormone levels. However, the injection of beta-Ep into human subjects resulted in a rise in plasma concentrations of prolactin. PMID- 6277127 TI - Radioimmunoassay detection of endorphins from long-term culture of human pituitary tumour cells. AB - Using a sensitive and precise radioimmunoassay for human beta-endorphin, we have demonstrated the sustained secretion of opioid peptides from human pituitary tumour cells. Pituitary tumour tissue obtained from a patient with Nelson's syndrome was maintained in continuous monolayer culture and secreted both beta lipotropin and beta-endorphin, with predominance of the latter. This is compatible with the idea that the beta-endorphin in normal human serum is secreted as such despite the predominance of beta-lipotropin compared with beta endorphin in the anterior pituitary. PMID- 6277128 TI - TSH and TSH-subunit production by human thyrotrophic tumour cells in monolayer culture. PMID- 6277129 TI - The effect of ACTH stimulation on plasma steroids in two patients with congenital hypoaldosteronism and in their relatives. AB - In two children with isolated congenital hyperreninaemic hypoaldosteronism, as well as in their relatives, plasma levels of aldosterone (Aldo), corticosterone (B), deoxycorticosterone (DOC), 18-OH-B and 18-OH-DOC were measured before and after an iv bolus of 0.25 mg Synacthen (Ciba). A corticosterone methyl oxidase deficiency type II was demonstrated in one child. Her normoreninaemic parents (no consanguinity) had plasma values consistent with heterozygosity. The results in the other child and one asymptomatic sib were compatible with a partial corticosterone methyl oxidase deficiency type I. His parents were consanguine but had normal Aldo levels. Overnight dexamethasone administration did not suppress any of the steroids measured except cortisol, suggesting synthesis of these steroids by the zona glomerulosa. PMID- 6277130 TI - Dexamethasone suppressible hyperaldosteronism in a child with nephrosclerosis. AB - A 9 year old Mexican boy presented with severe hypertension, hypokalaemia and features suggesting acute glomerulonephritis. Nephrosclerosis was present on renal biopsy. Aldosterone levels were unresponsive to variations in dietary salt intake and plasma renin activity was suppressed. Following oral dexamethasone therapy (2 mg/day), plasma aldosterone decreased to undetectable levels, serum potassium normalized and plasma renin activity gradually increased. Dexamethasone also restored the normal responsiveness of the renin-aldosterone system to postural stimuli. The patient exhibited a marked response to a single dose of ACTH with a rise in plasma aldosterone. Long-term blood pressure control and normal potassium levels have been achieved with oral prednisone therapy (5 mg/day) for a period of one year. This case of dexamethasone suppressible hyperaldosteronism (DSH) illustrates that the degree of hypertension in this syndrome may produce severe renal microvascular lesions. DSH should be considered in all children who present with low renin hypertension. PMID- 6277131 TI - Effects of ACTH on the zona glomerulosa of sodium-loaded timolol maleate-treated rats: stereology and plasma hormone concentrations. AB - The effect of chronic ACTH administration on the growth and aldosterone secretion of the zona glomerulosa of sodium-loaded timolol maleate-treated rats was investigated by stereological and radioimmunological techniques. Combined sodium loading and timolol maleate administration induced a significant atrophy of the zona glomerulosa and its cells. The plasma concentration of aldosterone was significantly decreased, while that of corticosterone did not vary. ACTH administration completely reversed zona glomerulosa atrophy, and the plasma concentration of corticosterone was noticeably enhanced, whereas that of aldosterone underwent a further decrease. This data suggests that in the absence of the other adrenoglomerulotrophic factors, ACTH stimulates the growth of the zona glomerulosa, but transforms its parenchymal elements to a functional fasciculata cell type. PMID- 6277132 TI - Interrelationship between calcium and cyclic nucleotides in platelet function. PMID- 6277134 TI - [Ultrastructural cytochemistry of periodic acid-reactive complex carbohydrates in human leukocytes (author's transl)]. PMID- 6277133 TI - Interaction of prostaglandin and cyclic AMP involved in regulation of platelet function. PMID- 6277135 TI - Infectious mononucleosis and Epstein-Barr virus in Japan. II. Cellular aspects. PMID- 6277136 TI - [Purine enzyme activities in lymphocytes from patients with impaired humoral immunity: decreased ecto-5'-nucleotidase activity in agammaglobulinemia (author's transl)]. PMID- 6277137 TI - Scanning electron microscopy of rat neurointermediate lobe cells in culture. AB - Enzymatically dispersed cells of the rat pars nervosa -- pars intermedia (NI) were observed by scanning electron microscopy after 1, 2 and 4 days in culture. The cells attached to the plate slowly, requiring about 3--4 days for the majority to adhere. The epithelial cells became attached singly and in clumps and branched chains, often over a bed of fibroblast-like cells. After the first day in culture, few surface features were evident on the NI cells, but by day 2, the surfaces showed a small number of blebs. In 4-day cultures, the cells revealed extensive areas with blebs and microvilli, and a few structures resembling cilia were observed. The adrenocorticotrophic hormone content of the cells after 4 days in culture was similar to that found in fresh tissue. PMID- 6277138 TI - Brindled mottled mouse: morphological changes of brain and visceral organs in hemizygous males following copper supplementation. AB - Intraperitoneal injections of cupric chloride prevent neuronal degeneration in the hemizygous brindled mottle mouse, MO br/Y, a murine model of kinky hair syndrome (KHS) in humans. At 6-9 months after two i.p. injections, the brain of MO br/Y revealed slightly increased amounts of lipofuscin pigments in the cerebral cortical neurons, cytoplasmic inclusions in the thalamic neurons, and axonal spheroid formation in the tuber cinereum, cerebellum and brain stem. Increased numbers of mitoses, bizarre hyperchromatic giant nuclei, and numerous clear vacuoles were frequently seen in the proximal renal tubular epithelium. Numerous myelin figures were conspicuous features in these epithelial cells at ultrastructural level. Such changes were not found in the littermate controls but in the heterozygous brindled mottled mouse, MO br/ +, identical changes were noted in equal or even higher frequency. These observations suggest that cupric chloride injections effectively modify the expression of the genetic defect in MO br/ Y. PMID- 6277139 TI - Unusual cytoplasmic bodies in Purkinje cells of the Gunn rats. AB - The fine structural changes of cerebellar Purkinje cells in Gunn rats were studied with electron microscope to clarify the neurotoxicity of bilirubin. The previous reports including mitochondrial change were confirmed. The new cytologic feature demonstrated in this study was the presence of two different types of unusual membranous cytoplasmic bodies (MCB). One type was the aggregation of rod like structures which were enveloped in double membranes and contained homogeneous and electron-dense matrix. Another type was the whirled lamellar structure which appeared to be formed of the microtubules and its unit particles. These two different MCB may suggest the different neurotoxicity of bilirubin: the former may result from the accumulation product of bilirubin-lipid complex, the latter may reflect the abnormal process of the microtubular assembly induced by bilirubin. PMID- 6277140 TI - N-Hexane- and methylethylketone-induced polyneuropathy. Abnormal accumulation of glycogen in unmyelinated axons. Report of a case. AB - This report presents the ultrastructural nerve study of a patient with sensorimotor neuropathy by a shoe glue containing an association of n-hexane and methylethylketone. Giant axons, distended by microfilamentous proliferation typical of such cases of neuropathy, were found in significant amounts. An unusual histological feature of this case was the considerable distension by a marked accumulation of glycogen granules in certain unmyelinated axons. Such a lesion has never been described in neuropathy induced by industrial agents. PMID- 6277141 TI - The localization of glial fibrillary acidic protein in brain tumors induced in hamsters with avian sarcoma virus. AB - The immunohistochemical localization of glial fibrillary acidic protein (GFAP) was studied by the peroxidase-antiperoxidase (PAP) method in avian sarcoma virus (ASV)-induced brain tumors in hamsters. One hundred twenty-four tumors including 54 astrocytomas, 64 pleomorphic gliomas, five sarcomas, and one unclassified tumor were stained with anti-GFAP serum. A positive immunostaining was seen in astrocytomas and in pleomorphic gliomas. Sarcomas and an unclassified tumor were negative. These results confirmed that a majority of ASV-induced brain tumors contained neoplastic glial cells, and further suggested that the astrocyte is a main target cell of ASV in the experimental neuro-oncogenesis. PMID- 6277143 TI - Filtration blebs in corneoscleral wounds sutured with Dexon 7-0 and Dexon 8-0. AB - In a retrospective investigation the incidence of post-operative subconjunctival filtration with bleb formation were studied in 112 eyes which had undergone cataract surgery with a corneoscleral incision sutured with braided polyglycolic acid sutures (Dexon). In 61 consecutive cataract operations sutured with Dexon 8 0 there were filtration blebs in 24 (39,3%) eyes, and in 51 consecutive cataract operations sutured with Dexon 7-0 there were filtration blebs in 11 (21,6%), eyes. Although there is a difference between the two materials, this is not statistically significant with the normal security level P less than 0.05. PMID- 6277142 TI - [Results of muscle biopsies in diphtheritic polyneuropathy. Light- and electron microscopic examinations of muscle fibers, intramuscular nerves, motor endplates, and intramuscular vessels]. AB - Muscle biopsies from the lower extremities of four patients with severe tetraplegic form of diphtheritic polyneuropathy were examined by modern techniques including histochemistry, electron microscopy and morphometric procedures. Until now comparable studies have not been published. The biopsies were removed during the acute stage of the polyneuropathy. We found scattered small angulated muscle fibers beside a more generalized slight atrophy predominantly of type 2 B fibers and targetoid-phenomenons or cores in type 1 fibers. Beside this neurogenic pattern there also were, corresponding with the results of electromyography, primary myogenic alterations with different degenerative phenomenons, suspicious of toxic origin as in cardiac muscle. The intramuscular vessels showed no abnormalities except some perivascular predominantly mononuclear cellular reactions with a remarkable number of cerebriform lymphoid cells, probably T-lymphocytes. No specific pathological alterations could be detected in 11 intramuscular nerves and two motor endplates. This may reflect the more proximal demyelination of human peripheral neurons by the diphtheria toxin as found in experimental diphtheria of the rabbit in contrast to the more distal type of the guinea pig. PMID- 6277144 TI - Central corneal thickness and intraocular tension in patients with acromegaly. AB - In 27 patients with pituitary adenomas the central corneal thickness and the intraocular tension were measured. Thirteen of the patients were suffering from acromegaly, and in this group the central corneal thickness was 0.561 mm +/- 0.35 (mean +/- SD). In the 14 patients with pituitary adenomas but no acromegaly the central corneal thickness was 0.526 mm +/- 0.030 (mean +/- SD). This difference is statistically significant 0.01 greater than P greater than 0.001. In the 13 patients with acromegaly the intraocular tension measured by applanation was 16.9 mmHg +/- 2.3 (mean +/- SD) and in the control group 14.7 mmHg +/- 2.4 (mean +/- SD). This difference is statistically significant (0.05 greater than P greater than 0.02). When the applanation reading is corrected for the difference in the central corneal thickness, the patients suffering from acromegaly have an intraocular tension of 14.1 mmHg, which is of the same magnitude as the tension in the patients without acromegaly. PMID- 6277145 TI - The bone inductive capacity of decalcified bone matrix modified by diphenylhydantoin. AB - Decalcified bone matrix was prepared from cortical bones of rats premedicated with I) Diphenylhydantoin (DPH), II) DPH + Vitamin D3, III) Vitamin D3 or IV) no premedication for 10 days. In the donor animals, DPH lowered the serum calcium level, caused a weight loss of 10 per cent, and stopped the growth of the long bones. Vitamin D3 supplementation normalized the serum calcium concentration but had no effect on the other parameters. Vitamin D3 alone caused hypertrophy of the growth cartilage, while the bone growth and structure was normal. The bone inductive capacity of decalcified bone matrix was highest in the DPH group, and the DPH + D3 group also showed significantly higher values than the D3, and control groups. The results of the present study show that the bone inductive capacity of the decalcified bone matrix is independent of Vitamin D3 metabolism. PMID- 6277146 TI - [Effect of Clerodendron bungei steud on electromyogram of ligament teres uteri in rabbits]. PMID- 6277147 TI - LH-RH induced changes in cAMP content of the anterior pituitary gland in male and female rats in vivo and in vitro. PMID- 6277148 TI - A comparison of experimental procedures of investigation of the dorsal root evoked ventral root reflex in the frog. AB - Effects of the drug methyl-m-aminobenzoate (MS-222 Sandoz) on the dorsal root evoked ventral root responses were studied by electrophysiological methods in the frog spinal cord. A fairly quick and marked depression of the response was observed from which complete recovery was seen within 60 minutes. Larger doses or repeated injection of small amounts of the drug prolonged the recovery and the monosynaptic discharge component of the ventral root reflex often deteriorated irreversibly. - In further experiments, monosynaptic ventral root discharges were demonstrated in spinal cords isolated from the vertebral canal and kept in Ringer solution. The results are discussed in the light of controversial views about the occurrence of monosynaptic ventral root discharge to stimulation of the primary afferents in the amphibian spinal cord. PMID- 6277149 TI - Effect of met-enkephalin and morphine on gastric secretion and blood flow. AB - The effects of met-enkephalin and morphine on gastric acid and pepsin secretion and gastric mucosal and total blood flow were studied in anaesthetized dogs with an in vivo chambered secretion stomach preparation. It was found that both agents infused intraarterially caused an increase in histamine-induced acid and pepsin secretion and mucosal and total blood flow. The above responses were significantly blocked by naloxone and nalorphine. In the resting stomach both opiates did not induce secretory changes but they increased mucosal and total blood flow. Met-enkephalin and morphine were also effective after intravenous administration. Met-enkephalin but not morphine fails to stimulate acid secretion if given into the portal vein. The likely mechanism of action of opiates on gastric secretion is discussed and a hypothesis of existence of opiate receptors in the gastric wall is presented. PMID- 6277151 TI - Incidence of hypothyroidism after 131I therapy for hyperthyroidism. Relation to pretherapy serum levels of T3, T4 and thyroid antibodies. AB - A correlation is reported between serum levels of triiodothyronine (S-T3) and thyroxine (S-T4) before treatment, as well as levels of thyroid antibodies before treatment, and the development of hypothyroidism following 131I therapy in 86 patients with hyperthyroidism. Patients with marked elevation of S-T3 or S-T4 had demonstrable antibodies to thyroid cytoplasmic antigen more often than those with normal or moderately elevated levels, and patients with markedly elevated levels of S-T3 also had a higher incidence of hypothyroidism after treatment. Patients with nodular thyroid glands and with markedly elevated levels of S-T3 required a larger number of 131I doses before no signs of hyperthyroidism persisted in comparison to those with moderately elevated levels. PMID- 6277150 TI - [Synthetic penicillins. Properties of new amidinopenicillins and their acetylmethyl esters]. PMID- 6277152 TI - Invasive squamous cell carcinoma of the uterine cervix. III. A malignancy grading system for indication of prognosis after radiation therapy. AB - The predictive value of a malignancy grading system embodied by 8 histopathologic parameters and of staging, age, year of admission and the different treatment modalities were compared in relation to 5- and 8-year survival. The material consisted of 196 patients in stage II according to the FIGO classification. The grading system proved superior with high significance (p less than 0.001) to the other predictors including each histopathologic parameter apart from vascular invasion (p = 0.01). All histopathologic parameters but 2 (differentiation into cell type and mitosis) exhibited significant (p less than 0.01) decrease in survival with higher grade at the 5- and 8-year controls. Patients in stage II could be divided into groups with low, intermediate and high malignancy points. Both groups of extremes showed a marked and highly significant difference in survival rates compared with the intermediate group, and the grading system made it possible to predict 60 per cent of the patients with acceptable specificity. In the group of high malignant tumours the system maintained a continual prognostic value even among patients still alive at the 5-year control. PMID- 6277153 TI - Advanced carcinoma of the nasopharynx. Treatment results. AB - From 1947 through 1977, 93 patients with carcinoma of the nasopharynx were treated. Most (83%) of the patients, had stage IV disease including 9 per cent who had a systemic tumor dissemination. Cervical lymph node metastases were present in 73 (78%) patients. The 5-year actuarial survival for all 93 patients was 13 per cent, 40 for stage II, 18 for stage III and 10 for stage IV patients. The factors with significant influence on survival were Initial Performance Status and T4 disease. The presence of N3 disease was a good indicator of a subsequent systemic tumor spread. PMID- 6277154 TI - Effects of vasopressin, noradrenalin and oxytocin on blood flow distribution in rat kidney with neoplasm. AB - The arterial blood flow distribution between tumour and intact renal tissue was investigated in rats with transplanted sarcomas. Changes in tissue flow were measured by the microsphere tracer technique, and the redistributing effects on blood flow of vasopressin, noradrenalin and oxytocin was recorded. Bolus injections of vasopressin gave a transient decrease of intact tissue flow, not found in tumours. At increasing doses of vasopressin and in early tumour growth phase, the flow discriminating effect tended to vanish. Constant intravenous infusion of vasopressin gave similar reduction of flow in tumour and intact tissue. Selective administration into the renal artery reduced flow in intact tissue but produced ambiguous effects in tumour tissue. Noradrenalin produced less reduction of tumour flow as compared with intact tissue flow. Oxytocin increased tumour blood flow while no flow change occurred in intact tissue. Oxytocin thus appeared to produce the most favourable redistribution of flow within tumour kidneys for the prospect of conveying cytotoxic agents selectively to the tumour. PMID- 6277155 TI - Broad-shouldered survival curves of a human melanoma xenograft. Implications for radiation therapy in the absence and presence of misonidazole. AB - A human malignant melanoma (E.E.) was irradiated in vivo in athymic nude mice, with subsequent assay of single cell survival in vitro. By assuming the survival curves for the hypoxic as well as for the aerobic cells in the tumour to be of the form S = exp(-D/D1) X (1-[1-exp (-D/D2]n), theoretic survival curves were fitted to the experimental data for tumours irradiated in air-breathing mice. Assumptions were made about hypoxic fraction, oxygen enhancement ratio, sensitization by misonidazole, repair of potentially lethal damage, and reoxygenation, all based upon own experimental data on E.E. melanoma. Theoretic surviving fractions were calculated for several clinically relevant fractionation regimes, both for irradiation in the absence and in the presence of misonidazole. The results indicate that tumours with biologic parameters like those of E.E. melanoma are best treated with fractionation regimes with few fractions and high doses per fraction, whether misonidazole is present or not. PMID- 6277156 TI - Malignant neoplasms among atomic bomb survivors following radiation therapy. AB - A second follow-up survey for radiation therapy exposure among a fixed cohort of A-bomb survivors originally numbering 20 000 persons identified 55 more who had received radiation therapy, and 7 more therapy-associated malignancies. In the previous and present investigations, 12 postirradiation malignancies were identified among 190 cohort members (6%) who received radiation therapy. This investigation underscored the necessity for documenting exposures to ionizing radiation for medical reasons, especially exposures incurring relatively high doses, among persons at relatively high risk of developing neoplasms, such as A bomb survivors, and in malignancy epidemiology to exclude bias in estimates of risk. PMID- 6277157 TI - Distribution of 35S in mice after oral administration of alpha dithioacetamidinium chloride. An autoradiographic investigation. AB - The distribution of 35S in mice has been investigated by whole-body autoradiography after oral administration of the 35S labelled S-75 (2,2' Dithiobis(N-[(1-adamantyl)-methyl]-acetamidine)dihydrochloride). The substance was rapidly absorbed, and the highest concentration occurred in the liver and kidneys. During the highest radiation protective activity (after about 45 min) a substantial concentration was found in the red pulp of the spleen. This supports previous findings that S-75 has its most marked protective effect on the splenic haemopoiesis. Previously, it was shown that cysteamine is more evenly distributed in the body than S-75, which is in agreement with its more generalized protective effect than S-75. PMID- 6277158 TI - Mammalian spermatogenesis as a biologic dosimeter for radiation. AB - Mouse spermatogenesis was used as an in vivo test system obviously suitable as a biologic dosimeter. Ionizing irradiation induced changes in the frequency distribution of the cellular DNA content of whole testis preparations. These changes can be analysed by flow cytometry. Such measurements deliver information on: (1) an increase of the coefficient of variation of the DNA histograms of some spermatogenic cells, (2) induction of diploid mature sperm, and (3) the dose depending inactivation of the highly sensitive differentiated spermatogonia. In this model especially No. 3 delivers quantitative information on the cytotoxic action of ionizing irradiation and chemical noxae as well, whereas Nos 1 and 2 may be used as qualitative criteria of mutagenic action of physical and chemical agents. No. 2 obviously being more sensitive than No. 1. PMID- 6277159 TI - Clonogenic potential of human tumors. A hypothesis. AB - Conventional radiation therapy of medium-sized (2 to 5 cm diameter) squamous cell carcinomas of the head and neck has resulted in data which were pooled to produce a curve of local control versus dose. From this curve, estimates were made of the number of clonogenic cells in these tumors. These calculations demonstrate a much smaller number of clonogenic cells than is usually assumed for human tumors. It is postulated that a relatively small proportion of the cells in human tumors are clonogenic. The number of cells which need to be destroyed by radiation therapy or chemotherapy in order to produce tumor control may be much less than usually estimated. PMID- 6277160 TI - Dissociation between changes in immunoreactive parathormone and its biological indices induced by cimetidine in primary hyperparathyroidism. AB - In three out of four patients with primary hyperparathyroidism, 2 000 mg of cimetidine daily caused a reduction of immunoreactive parathormone (iPTH) when measured at 8.30 and 11.30 on days 16 and 17 on treatment. Serum Ca, PO4 and maximal tubular reabsorption of PO4 remained unchanged. Excretion of cAMP/100 ml GFR remained elevated to at least the same extent as before treatment. Two patients, in whom cimetidine treatment was continued for an additional 4 weeks, did not show further hormonal or biochemical changes compared with the evaluation on days 16 and 17. We conclude that reduction of iPTH is not accompanied by any change in biological activity of this hormone. The reason for this discrepancy remains unclear. PMID- 6277161 TI - [Malignant retroperitoneal fibrous histiocytoma]. PMID- 6277162 TI - Afferent connections of the medial basal hypothalamus. PMID- 6277163 TI - Introduction to the role of psychosocial stressors in ischaemic heart disease. PMID- 6277164 TI - Molecular mechanisms of coupling in hormone receptor-adenylate cyclase systems. PMID- 6277165 TI - Mechanism of action of fructose 1,6-bisphosphatase. PMID- 6277166 TI - Immunity to parainfluenza type 3 virus and Pasteurella haemolytica in sheep. PMID- 6277167 TI - The ontogeny of the ruminant immune system and its significance in the understanding of maternal-fetal-neonatal relationships. AB - Considerable information is now beginning to accumulate on the ontogeny of immune responses in the ovine fetus. The fetal lamb has served as an important model for much of our present knowledge. Many critical questions remain to be answered in the ruminants. Much of this information provides a basis for a better understanding of the susceptibility of the fetus, the character of lesions and the events leading to persistent infections in congenital infections. The role that hormones associated with pregnancy plays in active and passive immunity in maternal, fetal and neonatal systems needs further definition. An understanding of these may make it possible to assist in reducing infections leading to congenital or neonatal disease. PMID- 6277169 TI - Taurine receptors in CNS membranes: binding studies. PMID- 6277168 TI - Interaction of taurine with its precursor, cysteine sulfinic acid. In the central nervous system. PMID- 6277170 TI - Studies on the renal handling of taurine: changes during maturation and after altered dietary intake. PMID- 6277171 TI - Recent advances in management of lung cancer. PMID- 6277172 TI - Radionuclide detection of gastrointestinal bleeding. PMID- 6277173 TI - Toxicology, industrial hygiene and medical control of TDI, MDI and PMPPI. AB - This presentation is authored particularly for those who demand comprehensive information regarding physiologic effects, toxicity, industrial hygiene and medical control of those isocyanates generally associated with polyurethanes. As a standard practice, every user, handler, storer, applicator, distributor or others who may come in contact with TDI, MDI and PMPPI should be aware of toxicologic risks associated with their misuse. The characteristic and most significant physiologic effect of exposure to these isocyanates is a direct irritant or sensitization response not unlike asthma. It is most unlikely that long-term exposure to TDI, MDI and PMPPI vapors would produce oncogenic effects. PMID- 6277174 TI - Radionuclide angiocardiographic assessment of pulmonary vascular reactivity in patients with left to right shunt and pulmonary hypertension. AB - Radionuclide angiocardiography was used to assess pulmonary vascular reactivity in eight patients (nine studies) with a large, relatively unrestrictive intracardiac defect and pulmonary arterial hypertension. Radionuclide angiocardiograms, using technetium-99m pertechnetate, were performed first with the patient breathing room air and then after 10 minutes of breathing a mixture containing 90 percent or more of oxygen. The pulmonary to systemic flow ratios obtained by gamma variate analysis of the radionuclide time-activity curves were compared with those calculated with the Fick principle at the time of cardiac catheterization. There was a good correlation between the two methods both in room air studies (r = 0.88) and in those obtained with 90 percent or more of oxygen (r = 0.94). All six studies (in five patients) with a reactive pulmonary vasculature (judged by a pulmonary vascular resistance at cardiac catheterization of less than 6 units/m2 with oxygen or after tolazoline) had a radionuclide pulmonary to systemic flow ratio of 3.0 or greater with oxygen. The three patients with a nonreactive pulmonary vasculature had a radionuclide pulmonary to systemic flow ratio of 2.3 or less with oxygen, a value that was unchanged from the room air value. These data suggest that radionuclide angiocardiography may be a useful, relatively noninvasive method of assessing pulmonary vascular reactivity in patients with a large, relatively unrestrictive intracardiac defect. PMID- 6277175 TI - Differential cardiovascular effects of calcium channel blocking agents: potential mechanisms. AB - The three major calcium channel blocking agents, diltiazem, nifedipine and verapamil, inhibit calcium entry into excitable cells. Despite this apparent common action at the cell membrane, these drugs produce quantitative and frequently qualitative differences in cardiovascular variables (for example, heart rate, atrioventricular [A-V] conduction and myocardial inotropic state) when evaluated at equieffective vasodilator doses. All three drugs increase coronary blood flow in a dose-dependent fashion (nifedipine greater than diltiazem = verapamil), and produce a negative inotropic effect in vitro in isolated atria and ventricles, also in a dose-dependent manner (verapamil greater than nifedipine greater than diltiazem). However, in conscious dogs nifedipine increases, verapamil decreases and diltiazem has little effect on the inotropic state. A-V conduction is slowed by diltiazem and verapamil but not by nifedipine in anesthetized dogs and in conscious dogs as judged from the P-R interval in the electrocardiogram. Heart rate is slowed in pentobarbital-anesthetized animals but is accelerated in conscious dogs (nifedipine greater than verapamil greater than diltiazem). Nifedipine also appears to interfere significantly with the arterial baroreceptor reflex by an apparent vagolytic action that is less evident with diltiazem and verapamil. Diltiazem, and possibly verapamil and nifedipine as well, appears to retard myocardial damage that accompanies ischemia. The mechanisms and sites of action of these drugs are presumed to be at the cell membrane; however, intracellular sites may also be involved. PMID- 6277176 TI - Selectivity of calcium antagonistic action in vascular smooth muscle. AB - Various mechanisms can account for selective inhibition by calcium influx blocking drugs of smooth muscle contractile activity. Diltiazem inhibits calcium influx activated by membrane depolarization as well as alpha adrenergic receptor stimulation. Diltiazem does not inhibit intracellular calcium release nor cause net calcium extrusion up to concentrations of 10(-5) M. As with other calcium influx blocking agents, diltiazem is a potent inhibitor for modes of activation that depend entirely on the presence of extracellular calcium. The calcium influx was found to be mediated by at least three different pathways: one activated by depolarization and highly sensitive to D-600, a second activated by norepinephrine and less sensitive to D-600, and a third "passive leak," that is insensitive to D-600 and accounts for calcium permeability in the resting state. Molecular differences among these various calcium channels may provide the basis for specificity of action of the calcium influx blocking agents. PMID- 6277178 TI - Selective action of diltiazem on cerebral vascular smooth muscle in the rabbit: antagonism of extrinsic but not intrinsic maintained tone. AB - Diltiazem selectively reduces the second equilibrium phase of contraction of several arteries and veins in the rabbit to a number of agonists, including norepinephrine, serotonin, histamine and potassium. Its effect is consistent with blockage of the entry of activator calcium from the extracellular space into the vascular smooth muscle cell. It has a preferential action on responses of the basilar artery in comparison with those of the ear and mesenteric arteries and saphenous vein. Although the spontaneously developing, maintained intrinsic tone of the basilar artery and facial vein are very resistant to diltiazem, the spontaneous rhythmic activity of the portal vein is sensitive to its antagonistic action. Findings reported here are consistent with the possibility that diltiazem preferentially blocks receptor-operated and potential-sensitive slow calcium channels of the cerebral artery compared with other systemic vessels tested but has little effect on the mechanisms responsible for maintained intrinsic tone development. PMID- 6277177 TI - Differential effects of calcium channel blocking agents on oxygen consumption rate in vascular smooth muscle. PMID- 6277179 TI - Ionic events responsible for the cardiac resting and action potential. AB - The cardiac action potential is distinguished from other excitatory phenomena by a prominent plateau and by the latent pacemaking capability of cardiac muscle. A review of experimental data suggests that ionic fluxes through gated membrane channels are the primary determinants of the shape of the cardiac action potential. The rapid depolarization phase of the action potential is mediated in part by an ionic channel that resembles the sodium ion (Na+) channel of nerve. A slower channel capable of carrying both calcium ion (Ca2+) and Na+ currents (Isi) also contributes to the upstroke of the action potential. The Ca2+ current through this channel is partly responsible for maintaining the plateau phase of the action potential. Moreover, because this slower channel activates at more positive potentials in partly depolarized myocardium or in specialized conduction tissue such as the sinoatrial and atrioventricular nodes. Two distinct transport systems appear to be the principal regulators of potassium ion (K+) in myocardium. An inwardly rectifying, voltage-dependent K+ channel apparently maintains all K+ conductance at rest and at all potentials negative to --20 mV. A second channel, which is both voltage- and time-dependent, evidently mediates K+ flux during the plateau phase of the action potential. This K+ current activates at potentials positive to --20 mV. The role of coupled transport mechanisms is now well established. The low intracellular concentrations of Na+ and Ca2+ prevailing in the myocardium are maintained by an electrogenic Na+ pump and a Na+ Ca2+ counter-transport system. Current data on carrier-mediated transport systems are insufficient to delineate the role of such mechanisms in the control of cardiac action potential. Further studies are required to provide details of the voltage, temporal and ionic dependence of gated channels, as well as of ionic counter-transport carriers, so that a quantitative reconstruction of cardiac action potential may be attempted. PMID- 6277180 TI - Effect of diltiazem on calcium transport and development of tension in heart muscle. PMID- 6277181 TI - Do calcium-dependent ionic currents mediate ischemic ventricular fibrillation? AB - Calcium ions mediate the adverse effects of myocardial ischemia and have been implicated in the genesis of arrhythmias. Calcium influx blocking drugs protect against early ventricular arrhythmias during experimental coronary occlusion, and recent studies suggest that this effect is at least partly due to inhibition of myocardial cell calcium influx. Most of the pharmacologic maneuvers used to simulate acute ischemic arrhythmias in vivo also produce intracellular calcium overload. Production of calcium overload in small myocardial cell clusters causes fibrillatory electrical and mechanical activity similar to that recorded from fibrillating hearts. Fibrillation in these cell clusters is mediated not by reentrant conduction, but by the same subcellular processes that give rise to depolarizing afterpotentials and abnormal automaticity. Agents favoring calcium influx, such as beta adrenergic agonists, accentuate these processes, while agents that depress calcium influx inhibit them. Although the relation of these experimental models to clinical ischemic arrhythmias has not been fully delineated, calcium influx blocking drugs may prove useful in reducing the incidence of sudden cardiac death. PMID- 6277182 TI - Comparative clinical electrophysiologic effects of diltiazem, verapamil and nifedipine: a review. AB - The slow channel blocking agents--diltiazem, verapamil and nifedipine--have generated clinical interest for the treatment of a variety of cardiovascular disorders. These agents, despite a similar basic mechanism of action, produce disparate clinical cardiac electrophysiologic effects in human beings. In usual doses, the acute administration of diltiazem slows heart rate. Verapamil and nifedipine, however, increase heart rate. Although diltiazem and verapamil produce equivalent slowing of atrioventricular (A-V) nodal conduction, verapamil prolongs A-V nodal refractoriness to a greater degree. In contrast, nifedipine facilitates A-V nodal conduction and shortens A-V nodal refractoriness. Knowledge of these differences may aid in the appropriate selection of specific slow channel blocking agents in specific clinical situations. PMID- 6277183 TI - Granular cell tumor of the esophagus. AB - Two cases of granular cell tumor of the esophagus are reported and the main features of the previously reported cases are summarized. Dysphagia and substernal discomfort or pain are the most common symptoms seen and are likely to occur with lesions greater than 1 cm. in diameter. The diagnosis should be considered in adult females with an intramural mass of the esophagus. The cell of origin is still disputed. The treatment of choice, when the patient is symptomatic or the lesion greater than 1 cm. in size, is local resection. The tumor, when incidentally discovered in an asymptomatic patient, may safely be followed endoscopically. PMID- 6277184 TI - Norwalk gastrointestinal illness: an outbreak associated with swimming in a recreational lake and secondary person-to-person transmission. AB - An outbreak of gastrointestinal illness in which headache, low grade fever and myalgia were common symptoms occurred among persons who visited a recreational park in Macomb County, Michigan, on July 13-16, 1979. The temporal clustering of onsets of 121 persons who were the first in their households to become ill suggested an incubation period ranging from 4-77 hours. A history of swimming in the park's lake was elicited with significantly greater frequency from these persons than from park visitors who were not ill (age standardized odds ratio = 4.8; 95% confidence interval, 1.8-12.7). One hundred twenty-six park visitors who became ill were household contacts of index patients who had swum in the lake; at least 62 of these 126 cases were probably due to secondary transmission. A secondary attack rate of 19% was observed in household contacts who had not visited the park. Serologic studies identified Norwalk virus as the etiologic agent. The source of the contamination of the lake could not be determined. Although some water samples collected just before and after the epidemic period had high coliform counts, the geometric mean coliform density of all samples collected on those days was within the limits established by the Environmental Protection Agency as acceptable for recreational contact water. PMID- 6277185 TI - Norwalk virus enteric illness acquired by swimming exposure. AB - In an epidemic of gastrointestinal illness strongly associated with swimming at a recreational park in Macomb County, Michigan, in July, 1979, the authors demonstrated the value of serologic testing to detect Norwalk virus infection. Rises in antibody titer to Norwalk virus were noted in all 11 individuals tested. Electron microscopy on stools from 20 ill individuals revealed only one with Norwalk virus-like particles. This particle was shown by radioimmunoassay and immune electron microscopy not to be Norwalk virus and not to have stimulated detectable antibodies in this individual. These results not only indicate that electron microscopy is insensitive in detecting Norwalk virus, but that it has the potential to mislead. A low rate of respiratory symptoms was associated with gastrointestinal illness in this Norwalk virus outbreak. The route of exposure might have been important for this. The outbreak was also noteworthy in that, although there was evidence of familial clusters of resistance, a very high percentage of the population was proved to be susceptible to the Norwalk virus. PMID- 6277186 TI - Foodborne Norwalk virus. AB - On December 6, 1979, three luncheon banquets were served in a New Jersey restaurant. Thirty-eight of 41 members (92.7%) of the first group became ill as did 25 of 31 members (80.6%) of the second group. None of 12 members of the third group were ill. Illness consisted primarily of diarrhea (76%), nausea, (73%), vomiting (67%), cramps (46%) and fever (18%); the median incubation period was 31 hours and median duration 24 hours. The same foods were served to all three groups, except that cole slaw was substituted for a green salad and mixed vegetables for lima beans for the third group. Consumption of green salad was associated with illness (p less than 0.0001). A total of 118 other persons who ate at the restaurant on the same day were interviewed, 60% of whom reported being ill. Green salad was significantly associated with illness for this group at lunch (p = 0.005) and dinner (p = 0.00007). Serologic studies on seven of 12 patients and on one of four exposed controls showed a fourfold or greater rise in antibody titer to Norwalk virus. PMID- 6277187 TI - The middle molecule hypothesis in perspective. AB - The middle molecule (MM) hypothesis states that molecules in the molecular weight range of 500 to 2000 daltons/molecule accumulate in uremia and are one cause of peripheral neuropathy. Evidence for and against this hypothesis is reviewed and evaluated. The course of events in the core of early hemodialysis patients who developed neuropathy provides important support ofr the hypothesis. Failure of early peritoneal dialysis patients to develop neuropathy suggested that the peritoneum is more permeable to MM's than early hemodialysis membranes, a fact that was later hemodialysis membranes, a fact that was later confirmed by appropriate investigations. Several investigators have demonstrated that MM's actually do accumulate in uremia and disappear rapidly following a renal transplant, which suggests a high clearance by the human kidney. Retrospective and prospective studies have demonstrated a protective effect of residual renal function against MM intoxication. Our prospective studies to produce MM intoxication are reviewed and their strengths and weaknesses delineated. Similarly, the investigations of others that are cited as evidence against the MM hypothesis are reviewed. It is the opinion of hte authors that the weight of evidence supports the validity of the MM hypothesis; but that final proof still is lacking. A protocol for more definitive studies is discussed. PMID- 6277188 TI - VP-16-213 (etoposide): the mandrake root from Issyk-Kul. AB - VP-16-213 (etoposide) is an orally and parenterally active antineoplastic agent synthesized from podophyllum extracts of a common North American plant, the May apple or mandrake. The drug delays and kills cells in the G2 phase of the cell cycle and is active in a variety of animal tumors and leukemias. Major therapeutic activity for the drug has been found in small cell bronchogenic carcinoma, germ cell malignancies, acute non-lymphocytic leukemia, Hodgkin's disease and non-HOdgkin's lymphoma. Minor therapeutic activity of the drug occurs in non-small cell bronchogenic carcinoma, pediatric neoplasms, hepatocellular carcinoma and gastric cancer. Toxicity is primarily hematologic, with alopecia, nausea and vomiting occurring less frequently. PMID- 6277189 TI - A variant of adrenomyeloneuropathy with hypothalamic-pituitary dysfunction and neurologic remission after glucocorticoid replacement therapy. AB - Adrenomyeloneuropathy is a syndrome comprising spastic paraparesis, polyneuropathy, primary adrenocortical insufficiency and variable hypogonadism. We describe a 32 year old man who presented with contractures, peripheral neuropathy, primary adrenocortical insufficiency adn secondary hypogonadism. Abnormal responses of growth hormone, gonadotropins, prolactin and thyrotropin to provocative stimuli were found, without radiographic evidence of a pituitary or hypothalamic lesion. Almost complete recovery from the neurologic abnormalities occurred with glucocorticoid replacement therapy. The clinical features of this patient support a diagnosis of adrenomyeloneuropathy. The hypothalamic-pituitary dysfunction extends the clinical features of this patient support a diagnosis of adrenomyeloneuropathy. The hypothalamic-pituitary dysfunction extends the clinical spectrum of this disease. Remission of the paraparesis coincident with glucocorticoid replacement has not been reported previously. PMID- 6277190 TI - Effects of low-calcium diet on urine calcium excretion, parathyroid function and serum 1,25(OH)2D3 levels in patients with idiopathic hypercalciuria and in normal subjects. AB - We have used a low-calcium diet providing only 2 mg/kg (body weight) per 24 hours of calcium to distinguish between "renal" and "absorptive" idiopathic hypercalciuria. Sixteen of 27 hypercalciuric subjects excreted calcium in excess of intake during days seven, eight and nine of he diet, suggesting some element of renal hypercalciuria; however, all patients had low or normal serum PTH and urine cAMP levels. In general, fasting urine calcium was elevated in these 16 subjects and normal in the remaining 11, who conserved calcium more normally. SErum 1,25(OH)2D3 levels were the same in patients and normal subjects, even though PTH levels of the patients were below those of he normal subjects. Urine magnesium excretion and phosphorus excretion were both increased in the patients who excreted calcium in excess of intake. Our findings suggest that renal and absorptive hypercalciuria may not be distinct entities but rather the two extremes of a continuum of behavior. A uniform elevation of intestinal calcium absorption and a variable defect of renal calcium reabsorption could explain our results far better than the hypothesis of distinct absorptive and renal forms of hypercalciuria. PMID- 6277191 TI - Immunology of sarcoidosis. PMID- 6277192 TI - Disorders of distal nephron function. AB - In this review, the distal nephron is considered to be that portion of the renal tubule commencing with the thick ascending limb of the loop of Henle and ending with the papillary collecting duct. The collecting duct, including its subdivisions in the cortex and medulla, originates from a different embryologic anlage than more proximal nephron segments, which may explain its morphologic and functional dissimilarities from the thick ascending limb and the distal convoluted tubule. This review summarizes selected aspects of the physiology of the distal nephron, with particular emphasis on the physiology of distal nephron transport of sodium, potassium, chloride and hydrogen ion. The pathophysiologic features of the following disorders of distal nephron function are reviewed: (1) pseudohypoaldosteronism, a heterogenous group of disorders in which the signs and symptoms are suggestive of aldosterone deficiency, but in which aldosterone levels are supernormal and administration of exogenous mineralocorticoid is not ameliorative; (2) pseudohyperaldosteronism (Liddle syndrome), a familial disorder in which the clinical manifestations closely resemble those resulting from an aldosterone-producing adenoma of the adrenal gland (primary aldosteronism), but in which the measured rate of aldosterone secretion and excretion is greatly subnormal; (3) Bartter syndrome and related syndromes of renal potassium wasting; (4) type 1 renal tubular acidosis (classic, distal); (5) type 4 renal tubular acidosis (hyperkalemic). Reference citations are generally to articles reporting recent advances in these areas and to review articles that contain comprehensive bibliographies. PMID- 6277193 TI - Control mechanisms in platelet activation. AB - Those activation stages that are energy dependent rely upon a complex control mechanism involving the interaction of platelet thrombasthenin, free ionic calcium, cyclic AMP and the prostaglandins. This article describes the feedback mechanisms inherent in these controls, and the sources, activities and degradation of all control materials. Particular attention is paid to the prostaglandins and their activating enzymes, both within the dense tubular system of the platelet and the cytoplasm of endothelial cells. The implications of the prostaglandin pathway for both therapy and laboratory diagnosis are explored. PMID- 6277194 TI - Criteria toward the definition of nonmetastatic gestational trophoblastic disease after hydatidiform mole. AB - The administration of chemotherapy to a patient with persistently elevated titers of hCG without metastases after hydatidiform mole implies a diagnosis of nonmetastatic gestational trophoblastic neoplasia (NMGTN). At present, a diagnosis of NMGTN following evacuation of hydatidiform mole is usually made after a plateau of three values of hCG over 2 weeks (days x, x + 7, x + 14), and patients are given chemotherapy on the basis of such a plateau. The frequency of the diagnosis of NMGTN from four United States Centers is presently 26% and this compares with a frequency of choriocarcinoma of 16% in the prechemotherapy era. Data are presented to demonstrate that a plateau of 3 or 4 weeks may be justified before a diagnosis of NMGTN is made and chemotherapy is given in patients who may be followed up closely. PMID- 6277195 TI - Immunologic and biologic characteristics of human decidual prolactin. AB - An increasing and consistent production of prolactin by decidual tissue (dhPRL) has been shown during a 24-hour incubation in a chemically defined buffer medium. No difference in dhPRL release was observed in tissue obtained from first- and third-trimester pregnancy. Estradiol had neither a stimulatory nor an inhibitory influence on dhPRL release, although release of this polypeptide by decidual tissue incubated with cycloheximide was blocked. The precise mechanism controlling the production and release of dhPRL is not known. Addition of theophylline, dibutyryl cyclic adenosine monophosphate, or guanosine triphosphate had no effect on dhPRL release during a 4-hour incubation. In contrast, these substances provoked a statistically significant increase in PRL release from rat hemipituitaries coincubated in vitro. Displacement curves for dhPRL against a human PRL standard suggest molecular homogeneity between both hormones. The biologic activity of dhPRL was next confirmed by means of a new bioassay system with a lower level of sensitivity of 20 pg/ml. In addition, dissociation of the biologic and immunologic activity of dhPRL has been completed and suggests that at least three isohormones are produced by human decidual tissue. The dhPRL production rate per gram of decidual tissue wet weight is far below that reported for the pituitary homologue. These studies not only provide confirmatory evidence of the immunologic and biologic activity of dhPRL but also intensify speculation as to its role in human gestation, including that related to osmoregulation across human amnion. PMID- 6277196 TI - The effects of estrogen on adrenal androgens in oophorectomized women. AB - The serum levels of adrenal androgens (aa) are lower in oophorectomized (OO) than in ovulating (OV) women. This study was carried out in an effort to further investigate these findings and to study the effects of administration of estrogen on the levels of aa in OO women. Ten OO and seven OV women participated in this study in which aa were measured basally and after stimulation with adrenocorticotropic hormone (ACTH), both before and 4 weeks after conjugated estrogens (CE). Seven women received 0.625 mg of CE, and five received 2.5 mg of CE. Compared to OV women, OO women had significantly lower levels of androstenedione (Adione), dehydroepiandrosterone (DHEA) and its sulfate (DHEA-S), testosterone (T), delta 5-androstenediol (Adiol), and 17 beta-estradiol (E2) (p less than 0.01). In response to ACTH, OO women had smaller responses to Adione (p less than 0.05), DHEA (p less than 0.005), DHEA-S (p less than 0.01), 17-OH progesterone (17 Prog) (p less than 0.01), and 17-OH pregnenolone (17 Preg) (p less than 0.1). Furthermore, after ACTH, the ratio of 17 Prog/Adione was significantly higher in OO women (p less than 0.01), thus suggesting reduced 17,20-demolase (17,20D?) activity. Similarly, OO women had higher ratios of 127 Preg/17 Prog (p less than 0.1), DHEA/Adione (p less than 0.01), and Adiol/T (p less than 0.01), thereby suggesting reduced 3 beta ol dehydrogenase-isomerase (3 beta ol) activity. In response to CE, there was a dose-related increase in aa and cortisol. After 2.5 mg of CE, aa were significantly higher and similar to those levels in OV women. despite the known increases in sex hormone binding globulin finding capacity and transcortin after estrogen, unbound T increased slightly, as did urinary free cortisol in women treated with 2.5 mg of CE. After treatment with estrogen, there was a dose-related change in the ACTH-stimulated steroid ratios that indicated and increase in 17,20D and 2 beta ol activities. In women who were gien 2.5 mg of CE, these enzyme activities were similar to those in OV women. PMID- 6277197 TI - The nature of trophoblastic disease initiated by transplantation into immunosuppressed animals. PMID- 6277198 TI - Inhibiting herpes simplex virus type 2 infection in human epithelial cells by gossypol, a potent spermicidal and contraceptive agent. PMID- 6277199 TI - Effects of a thioreactive agent, diamide, on neuromuscular transmission in lobster. AB - Diamide[diazine-dicarboxylic acid-bis(dimethylamide)], a thiol-oxidizing agent, has both pre- and postsynaptic actions on the glutaminergic neuromuscular junction of the lobster walking leg. Postsynaptically, diamide produced an increase in the response to exogenously applied glutamate, whereas the effect of diamide on presynaptic transmitter release involved two major changes: 1) a decrease in excitatory junction potential amplitude and 2) an increase in miniature junction potential frequency. Short-term facilitation also decreased. Equilibration with 1,4-dithiothreitol (a sulfhydryl-reducing agent) reversed the decline in excitatory junction potential frequency, and the fall in short-term facilitation. The miniature junction potential frequency increase induced by diamide was independent of external Ca2+, as diamide in a Ca2+-free solution produced a similar response to that in a Ca2+-containing solution. We propose that the action of diamide on transmitter release is similar to the action of polyvalent cations, i.e., diamide has two sites of action, a blockade of inward Ca2+ flux and an increased release of Ca2+ inside the terminal. PMID- 6277201 TI - Evidence for two distinct modalities of CA2+ influx into pancreatic B cell. AB - The pathways through which glucose stimulates Ca2+ inflow into islet cells were investigated by comparing the inhibitory effect of verapamil, a selective blocker of voltage-sensitive Ca2+ channels, on glucose- and K+-stimulated insulin release and 45Ca efflux from perifused rat pancreatic islets. The islets stimulated by K+ (20 mM) were more sensitive to verapamil than those exposed to glucose (27.8 mM). The stimulation of 45Ca efflux by a low concentration of glucose (8.3 mM) was extremely resistant to verapamil, whereas that induced by a rise in the glucose concentration from 8.3 to 27.8 mM displayed the same sensitivity towards verapamil as that characterizing the response to K+. Because the increase in 45Ca efflux evoked by glucose or K+. Because the increase in 45Ca efflux evoked by glucose or K+ reflects a stimulation of Ca2+ entry into islet cells, it is proposed that the B cell may be equipped with two populations of Ca2+ channels that differ in their sensitivity towards verapamil and possibly their voltage dependency. Glucose apparently stimulates Ca2+ inflow through both types of channels. At low concentrations, glucose may stimulate Ca2+ inflow, in part, by gating the voltage-insensitive pathway. PMID- 6277200 TI - Development of Na+-dependent hexose transport in a cultured line of porcine kidney cells. AB - LLC-PK1 cells in culture do not concentrate alpha-methylglucoside (alpha-meG) during their early growth phase but develop the capacity to concentrate this hexose as the growth rate decreases in confluent cultures. The concentrating ability is dependent on the Na+ electrochemical gradient and is inhibited by phlorizin with KI,0.5 approximately 0.2 microM. The development of the concentrative capacity can be accelerated by the Friend cell inducer hexamethylene bisacetamide (HMBA) and by the phosphodiesterase inhibitors dibutyryl cAMP, theophylline, and 1-methyl-3-isobutylxanthine (MIX). In cultures treated with any of these differentiation-accelerating chemicals, the development of alpha-meG concentrating capacity is severely inhibited by the tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA) but not by inactive (in tumor promotion) analogs of TPA. In all cases, an early event in the development of alpha-meG accumulating capacity is an elevated intracellular cAMP concentration; however the results suggest that this increase in cAMP may be necessary but not sufficient to induce the differentiated hexose-accumulating capacity. PMID- 6277202 TI - Studies of the urinary acidification defect induced by lithium. AB - Experiments were carried out in rats and isolated turtle bladders to study the defect in H+ transport induced by LiCl. After 3-4 days of intraperitoneal LiCl, rats developed urinary findings of "distal" renal tubular acidosis. Proximal tubular fluid pH measured in situ by glass microelectrodes was higher in lithium treated rats than in acidotic controls. Proximal fluid total CO2 [tCO2] was also higher, and the fraction of tCO2 leaving the proximal tubule was 14 vs. 7% (P less than 0.001). Impaired acidification was also apparent beyond distal convoluted tubules, as judged by normal distal tCO2 reabsorption but increased HCO3(-) in the urine. During NaHCO3 loading, the proximal defect was ameliorated but not the distal. Turtle bladder studies showed that mucosal lithium inhibits H+ secretion secondary to reducing transepithelial electrical potential, presumably by hyperpolarization of the luminal membrane. A similar mechanism may be responsible for lithium's effect on the distal nephron. Inhibition of proximal tubular HCO3(-) reabsorption is probably not attributable to electrical potential changes but might be due to interference of luminal membrane Na+ entry by Li+ and reduced (Na+ + Li+)-H+ exchange. PMID- 6277203 TI - Interaction of verapamil with human platelet alpha-adrenergic receptors. AB - Platelet aggregation induced by epinephrine is an alpha-adrenergic event and is inhibited in vitro by the calcium-channel blocker, verapamil. We wondered whether this inhibition might be mediated by an interaction of verapamil with platelet alpha-adrenergic receptors. Verapamil inhibited the specific binding of [3H]dihydroergocryptine ([3H]DHE), a nonselective alpha-adrenergic antagonist, to intact platelets in a concentration-dependent and competitive manner. At 10 microM verapamil caused a fivefold decrease in the affinity of binding of [3H]DHE to platelet alpha-adrenergic receptors (P less than 0.001) with no change in the number of receptors per platelet. The inhibition of binding was not reversed by excess calcium. Verapamil also inhibited the binding of a selective alpha 2 adrenergic antagonist, [3H]yohimbine, to intact platelets, and inhibited the binding of an alpha 2-partial agonist, [3H]clonidine, to platelet membranes. Moreover there was a strong correlation between verapamil's effect on [3H]DHE binding and its effect on epinephrine-induced aggregation (r = 0.98). Although verapamil is commonly used as an inhibitor of calcium flux across cell membranes, the present studies demonstrate that verapamil also inhibits epinephrine-induced aggregation at the level of the platelet alpha-adrenergic receptor. PMID- 6277205 TI - Visual effects of a psychoactive short-chain neuropeptide. PMID- 6277204 TI - Direct measurement of sarcomere length from isolated cardiac cells. AB - Isolated cardiac muscle cells enzymatically digested with collagenase and hyaluronidase from whole rat myocardium demonstrate the characteristics of an intact membrane in that they tolerate millimolar concentrations of free Ca2+ and exhibit phasic contractions with electrical excitation. These isolated cells maintain their characteristic A-I band striation patterns when at rest or during contraction. An apparatus has been developed to directly image these cells with phase-contrast micrography onto a 1,728-element charge-coupled device photodiode array for rapid data storage in a digital computer. The digitized striation pattern profile was analyzed for individual and average sarcomere spacing. In isotonic media the average resting sarcomere length ranged from 1.77 to 1.91 micrometers in 13 cells, with a mean length of 1.83 +/- 0.12 micrometers. Electrically stimulated phasic contractions in three cells demonstrated a synchronous 20% decrease in sarcomere spacing to a mean of 1.51 micrometers. Striation spacing decreased under hypertonic stress but elongated only up to 1.93 micrometers in hypotonic solutions, suggesting that some internal elastic constraint exists that limits elongation of the cell. PMID- 6277206 TI - Human rotavirus infection in Efate, Vanuatu. AB - Serum and fecal samples collected from children with gastroenteritis and healthy children and adults living in Efate, Vanuatu (formerly the New Hebrides), were tested for the presence of human rotavirus antigen or antibody by electron microscopy and enzyme-linked immunosorbent assay. Virtually every subject was found to have detectable levels of antibody and age-specific studies showed that primary infections occur early in life. Human rotavirus was demonstrated to be the cause of an outbreak of gastroenteritis among children which occurred between August and September 1980, although it had not been detected in the population in the preceding 13 months. Epidemics of human rotavirus-associated gastroenteritis appear to occur every 2nd year in this population. PMID- 6277208 TI - Adenoid cystic carcinoma of the breast. A case report. PMID- 6277207 TI - The relative importance of rotavirus and other pathogens in the etiology of gastroenteritis in Trinidadian children. AB - Over a 2-year period, 300 infants less than 3 years old with gastroenteritis admitted to hospitals in Trinidad were investigated for the presence of certain microorganisms in the feces, along with an equal number of age- and sex-matched controls. Rotavirus was detected in 23% of cases and 1% of controls; Salmonella in 7% of cases and in 1% of controls; Shigella in 4% of cases and in no controls and two serotypes of enteropathogenic E. coli in 7% of cases and in 2% of controls. Campylobacter fetus subspecies jejuni was cultured from 7 out of 60 cases and from 1 of 60 controls. Enterotoxigenic E. coli, most strains of enteropathogenic E. coli, cytopathic enteroviruses and adenoviruses and fecal parasites were not significantly associated with diarrhea. Rotaviruses were detected throughout the year but were more prevalent in the dry than in the rainy season. They were found less often in children younger than 6 months than in those aged 6 to 35 months and were present in 6 of the 20 children who died. PMID- 6277209 TI - [Etomidate-infusion-anaesthesia. I. Haemodynamics and blood gases analyses in the early postoperative period (author's transl)]. AB - The influence of etomidate-infusion anaesthesia on blood pressure, heart rate and blood gases in the early postoperative period has been investigated on 14 patients. An infusion model for etomidate is described. Blood pressure and heart rate remained unaltered. There was only a max. fall of pO2 for 7.7% and a max. rise in pCO2 for 5.4%. Metabolic parameters did not change. PMID- 6277210 TI - [Cytomegalovirus infection. An obstetrical observation (author's transl)]. AB - Cytomegalovirus infections are transmitted among adults by transfusion of fresh blood. This benign infection is characterised by a single febrile syndrome that appears, on an average, 22 days after the transfusion and may last as long as 6 weeks. Hematological anomalies are significant: inversion of the leucocytic formula with high rate of mononucleosis, founded on the presence of hyperbasophilic mononuclear cells that appears about the 30th day, will be confirmed when the virus is isolated in mononuclear cells and in urine and by means of situation fo C. M. V. antibodies. Those post-transfusion C. M. V. infections are frequent after extra-corporeal circulation but only appear clinically in 1 p. cent of cases. PMID- 6277211 TI - Tissue reaction to suture materials in infected surgical wounds--a histopathologic evaluation. AB - Suture materials are foreign bodies which induce and prolong tissue reaction in the wound area. Cellular responses to infected wounds containing 7 suture materials (braided polyglycolic acid [BPGA, Dexon], multifilament stainless steel, monofilament nylon, black braided silk, braided Dacron [Mersilene], and plain and chromic catgut) were studied at 6, 10, 20, and 40 days. Cellular reaction varied with different suture materials. In general, neutrophils were the predominant cells in acute infection, but later, macrophages and fibroblasts predominated. Occasionally, plasma cells, lymphocytes, eosinophils, and giant cells were present. In the infected wounds with implants of plain catgut, chromic catgut, silk, and braided Dacron, there were large numbers of neutrophils even in chronic implantation, indicating persistence of local infection. With nylon, steel, and BPGA, the number of neutrophils rapidly decreased. Although BPGA induced intense acute reaction, the response was mild in chronic implantation. It appeared to be absorbed more slowly than plain or chromic catgut, and tissue reaction was minimal in the absorption stages. A reaction similar to the Splendore-Hoeppli phenomenon was seen around multifilament silk strands in chronic stages of implantation. PMID- 6277212 TI - Further studies with polyglycolic acid (Dexon) and other sutures in infected experimental wounds. AB - Seven suture materials (braided polyglycolic acid [BPGA, Dexon], multifilament stainless steel, monofilament nylon, black braided silk, braided Dacron (BD, Mersilene], and plain and chromic catgut) were implanted in surgical wounds which were inoculated with 3 dilutions of Staphylococcus aureus. The degree of swelling was measured at 6, 10, 20, and 40 days. Monofilament nylon and multifilament steel produced the least amount of swelling; plain and chromic catgut produced the most. The BPGA, BD, and silk caused swellings that were intermediate in degree. In chronic implantations, inflammatory reactions to the various suture materials differed from those occurring soon after implantation. Response to steel and nylon remained minimal; that to silk was gradual, but overall became the most severe. There was slightly less response with BD. The catguts varied little from each other in their responses. Intense reaction was induced by BPGA in the early stages, but the response was mild in chronic implantations. PMID- 6277213 TI - Immunogenicity of bivalent vaccine for reproductive failure of swine induced by pseudorabies virus and porcine parvovirus. PMID- 6277214 TI - Effects of incubation temperatures and bovine viral diarrhea virus immune serum on bovine turbinate cells persistently infected with a noncytopathogenic isolate of bovine viral diarrhea virus. AB - Noncytopathogenic bovine viral diarrhea virus (BVDV) on initial inoculation in receptive cells induced a persistence in vitro at incubation temperatures of 33, 37, and 39 C. The mechanisms of this persistence were not the result of the induction of defective interfering particles or the selection of temperature sensitive mutants, but were the result of a naturally occurring noncytopathogenic isolate of BVDV. Persistently infected cells were not freed of the infection by continuous passage in media containing homologous antibody. Persistently infected cells appeared to undergo an earlier senescence than did noninoculated cells of the same passage level and age. This phenomenon was reversed by renewal of the culture media. PMID- 6277215 TI - The effect of granulocyte transfusions on the incidence of cytomegalovirus infection after allogeneic marrow transplantation. AB - The association of granulocyte transfusions with cytomegalovirus infection and interstitial pneumonitis in allogeneic marrow transplant recipients was reviewed. Data from 387 patients with aplastic anemia or acute leukemia receiving transplants from human leukocyte antigen-matched siblings were analyzed. Acquisition of cytomegalovirus infection was higher in recipients of prophylactic and therapeutic granulocytes than in control subjects. The incidence was significantly different, however, only in cytomegalovirus seronegative patients who received granulocytes from seropositive granulocyte donors. There was no significant increase in the total incidence of or mortality from all forms of interstitial pneumonitis. PMID- 6277217 TI - Immune globulins for protection against viral hepatitis. Recommendations of the Immunization Practices Advisory Committee. Centers for Disease Control, Department of Health and Human Services, Atlanta, Georgia. PMID- 6277216 TI - Pulmonary eosinophilic granuloma: electron microscopic detection of X-bodies on lung lavage cells and transbronchoscopic lung biopsy in one patient. PMID- 6277218 TI - Pneumococcal polysaccharide vaccine. Recommendation of the Immunization Practices Advisory Committee. AB - The risk of pneumococcal infection differs in different age ranges; the risk and the severity of the infection increases in persons with certain chronic diseases. Penicillin remains the best antimicrobial agent for treatment. The recently developed pneumococcal polysaccharide vaccine represents 14 serotypes responsible for a large majority of pneumococcal infections in the United States. The efficacy of the vaccine has been demonstrated in a number of trials. this vaccine recommendation statement indicates the persons most likely to benefit from vaccination and reviews the adverse effects likely to occur and the precautions to be taken in administering the vaccine. PMID- 6277219 TI - Prophylactic granulocytes in the neutropenic host. PMID- 6277220 TI - Cortisol suppression test in patients with elevated adrenocorticotropic hormone levels. AB - Increased adrenocorticotropic hormone (ACTH) levels after bilateral adrenalectomy could be secondary to a pituitary tumor, under replacement with cortisol, or an abnormality in the hypothalamic-pituitary-adrenal feedback loop. To distinguish between these possibilities, ACTH levels were measured before and after cortisol infusion (20 mg/h for 4 hours) in five groups: normal volunteers; patients with idiopathic adrenal insufficiency; and with bilateral adrenalectomy for Cushing's syndrome with no roentgenographic evidence of pituitary tumor, with pituitary tumors, and with equivocal roentgenographic studies (suspect pituitary tumors). Control ACTH levels in all groups of patients were higher than in normal volunteers but there was overlapping. Cortisol infusion suppressed ACTH in all subjects but the reductions in the last two groups were less than in the first three. The cortisol suppression test appears to be useful in determining whether increased ACTH level after adrenalectomy is due to a pituitary tumor. PMID- 6277221 TI - Autonomic nervous system abnormalities and allergy. AB - Abnormal autonomic nervous system responsiveness may contribute to the pathogenesis of asthma and other allergic diseases. Therefore, we measured alpha- and beta-adrenergic and cholinergic responsiveness in allergic subjects. Allergic asthmatic subjects had an abnormal adrenergic (alpha = hyperresponsive; beta = hyporesponsive) and cholinergic (hyperresponsive) profile. However, subjects with allergic rhinitis and preallergic subjects (those with positive allergen skin tests without any disease manifestation) had equivalent beta-adrenergic and cholinergic abnormalities. Thus, all allergic subjects showed abnormal beta adrenergic hyporeactivity and cholinergic hypersensitivity whereas allergic asthma was singularly associated with excessive alpha-adrenergic responsiveness. Autoantibodies against beta-receptors were found predominantly in subjects with beta-adrenergic hyporeactivity. The presence of these autoantibodies and the physiologic abnormalities associated with their presence suggests a causitive relationship. PMID- 6277222 TI - [Intratypic serodifferentiation of poliovirus by immunodiffusion test (author's transl)]. PMID- 6277223 TI - Suspected epidemiological relationships among strains of Escherichia coli O55:B5 confirmed by restriction pattern analysis of the carried plasmid (RColBM IncFIII). AB - Seven strains of Escherichia coli O55:B5g, isolated from independent cases of infantile diarrhoea, contained similar RColBM plasmids beloning to incompatibility group IncFIII. Suspecting an epidemiological link among all these cases we compared the restriction pattern of the corresponding RColBM plasmids after digestion with endonucleases EcoRI and BglII. These patterns were similar, confirming the suspected relationships among the seven cases of infantile diarrhoea. The patterns obtained with the RColBM plasmids from children were different from those obtained with two RColBM plasmids isolated from adults. The probable source of the seven cases of infantile diarrhoea was identified as a nursery. PMID- 6277224 TI - [Necrotizing enterocolitis. A study of forty-six cases seen in a maternity hospital (author's transl)]. PMID- 6277225 TI - A case of insulinoma diagnosed by plasma insulin, selective pancreatic angiography and computerised tomography. AB - A 33 year old woman with benign islet cell adenoma in the head of the pancreas with hyperinsulinism is reported. The application of blood glucose levels with simultaneous plasma insulin assays, selective angiographic and computerised tomographic studies in its diagnosis and localisation of the tumour is discussed. After enucleation of the insulinoma, the patient recovered. PMID- 6277226 TI - Radiation induced cancer: a report of 10 cases. AB - Ionizing radiations have been shown to be carcinogenic to man as well as experimental animals. Malignancies following therapeutic radiation occur rarely. Over the past 10 years the authors recorded 10 cases of tumours in irradiated tissues. 3 occurred in patients irradiated for nasopharyngeal carcinoma, 3 were irradiated for tuberculosis adenitis, 2 for carcinoma of the cervix, 1 for carcinoma of the breast and 1 for basal cell carcinoma. The latent period for tumour induction following the irradiation varied from 5 years to 31 years. All these cases showed no evidence of recurrence or metastases of the original primary lesion; and the histology of the second primary differed from the first. Evidence of radiation damage was seen in all cases except for 2 patients who were treated for tuberculosis adenitis. The doses received varied from 900r to about, 9000r. Among the tumours produced, there were 3 cases of squamous cell carcinoma of the oral & postcricoid region, 2 cases of papillary carcinoma of the thyroid, 2 cases of adenocarcinoma of the rectum, 1 case of adenocarcinoma of the ethmoid, 1 case of osteosarcoma of the mandible and 1 case of extraskeletal osteosarcoma. The clinical features of these cases are discussed and other cases reported in the literature are reviewed. PMID- 6277227 TI - The molecular biology of cancer and its diagnostic implications. AB - The origin of cancer is discussed from the view of the two-stage model of malignant transformation. Environmental carcinogens play an integral part in the process. When the cell is transformed, cell surface changes are found for such components as fibronectin, collagen, actin, myosin, glycopeptides and enzyme activities. Hormone receptors are a fruitful line for research. Both qualitative and quantitative alterations are also seen with cancer cell enzymes. Among enzymes that can be used as markers of malignancy are the protease. A group of oncodevelopmental proteins, hormonal and non-hormonal, are in regular service for the management of cancer. Improvements in diagnostic specificity can be expected as the newer technologies are harnessed for medical use. PMID- 6277228 TI - The curability of advanced cancers with chemotherapy. AB - The tremendous progress that has been made in the chemotherapy of malignant diseases since the early 1950's has enabled the cure of a significant number of cancers such as chloriocarcinoma, Burkitt's lymphoma, Hodgkin's disease, non Hodgkin's lymphoma, the acute leukaemias, testicular carcinoma, and many childhood cancers such as rhabdomyosarcoma, Wilm's tumor, Ewing's sarcoma, ovarian cancer, and retinoblastoma. As a result, the mortality from cancers has dropped by 15% for persons under the age of 45 years and even more for those under 30 years of age. Many other metastatic cancers can now be successfully controlled with chemotherapy and, ultimately, more will be added to the growing list of curable cancers. The chemotherapeutic agents responsible for the cures of some cancers include asparaginase, actinomycin D, Adriamycin, bleomycin, cisplatin, cyclophosphamide, cytosine arabinoside, 5-fluorouracil, 6 mercaptopurine, methotrexate, nitrogen mustard, prednisone, procarbazine, and vincristine. The discovery of new effective drugs such as AMSA and anthracenedione promises to improve the success rates obtained with present therapy. Chemotherapy is indicated for every patient who has metastatic cancer, since virtually every patient can receive some palliation from such therapy, while for some patients chemotherapy holds the promise of prolongation of life or even cure. PMID- 6277229 TI - Methods of reduction of cisplatin nephrotoxicity. AB - Cisplatin, an agent widely used in the chemotherapy of a variety of human malignancies, is often dose-limited owing to its nephrotoxicity. Some of the approaches under consideration, regarding the reduction of cisplatin nephrotoxicity, include the use of hydration and osmotic diuresis, pharmacological diuretics, chelating agents or agents which otherwise react with cisplatin or reverse cisplatin-induced deoxyribonucleic acid cross-links, and antioxidants to destroy free radicals, especially superoxide radicals, produced by cisplatin. The effects of each of these and other interventions on cisplatin induced nephrotoxicity are delineated, along with their proposed mechanisms and effects on therapeutic efficacy. The current status of development of organoplatinum analogs yielding congeners with less nephrotoxicity and greater efficacy is discussed briefly. Finally, a possible role of endogenous and/or exogenous prostaglandins in protecting against or reversing heavy metal nephrotoxicity is suggested. PMID- 6277230 TI - Regional localization of the genes for human HEXB. PGK, GALA. HPRT, G6PD by somatic cell hybridization. AB - Twenty independent man-mouse (Cl1D,LA/TK-, HPRT-) and man-hamster (CH,HPRT-) hybrids using female human cells with balanced reciprocal translocation XX,t(X;5)(q21;q11) were analyzed for human genes localized on chromosome 5 (HEXB), on chromosome X (PGK, GALA, HPRT, G6PD) and for the different chromosomes in relation with the balanced reciprocal translocation (chr.5, chr.5q-, chr.Xq+, chr.X). The different results obtained indicate that the genes for human markers HEXB, PGK are on Xq+, and that the genes for human markers GALA, G6PD are on 5q-. These data implicate finally the following localizations: HEXB on 5q11 leads to 5qter; PGK on Xq21 leads to Xpter; GALA, HPRT, G6PD on Xq21 leads to Xqter. PMID- 6277232 TI - [Odontodysplasia. Report of a case (author's transl)]. PMID- 6277231 TI - Employment of toxins bound to porous silica beads for isolation of pure antibodies by immunoadsorption. AB - A simple technique for the isolation of pure antibodies by use of bacterial toxins or toxoids bound to porous silica beads "Spherosil" is described. Employment of toxins instead of toxoids has the advantage of resulting higher yields of purified antibodies. PMID- 6277233 TI - Value of C26:O fatty acid determination for the diagnosis of atypical adrenoleukodystrophy. PMID- 6277234 TI - Isolation of herpes simplex virus type I in recurrent (Mollaret) meningitis. PMID- 6277236 TI - Nerve stimulation test in murine experimental autoimmune myasthenia gravis. AB - Use of the mouse model of myasthenia gravis (murine EAMG), ideally suited for immunological study, has been hampered by the relatively mild character of the disease and by the extended time and effort required for inducing severe disease. Electromyographic measurement of the compound action potential after repetitive stimuli, the nerve stimulation test, was evaluated for its ability to diagnose neuromuscular transmission defects in mice immunized with Torpedo acetylcholine receptor. With the combination of provocative maneuvers and raising of the body temperature, EAMG could be diagnosed in nearly all immunized animals a few weeks after immunization, whereas clinical evaluation of muscle weakness was positive in less than half of immunized mice months after the first immunization. Thus, EMG provides a means of objective evaluation of the disease and attempts at its experimental modification. PMID- 6277235 TI - Interaction of myasthenic immunoglobulins and cholinergic agonists on acetylcholine receptors of rat myotubes. AB - Acetylcholine receptor (AChR) antibodies from patients with myasthenia gravis (MG) impair neuromuscular transmission by altering the number of AChRs at the skeletal muscle motor endplate. Cholinergic agonists similarly impair transmission by altering the number and affinity of AChRs. The frequent clinical resistance ot anticholinesterase medication in patients with MG has raised the question of possible synergistic effects of these agents on regulation of AChRs and neuromuscular transmission. To investigate this question, rat myotube cultures were incubated with MG globulin and carbamylcholine (CMC) for 20 hours. The number of AChRs was assayed by 125I-alpha-bungarotoxin binding. Incubation of cultures with both MG globulins and CMC consistently produced greater reductions of AChRs than incubation with either substance alone. Cholinergic antagonists blocked the CMC effect but not the MG globulin effect. A muscarinic antagonist had no effect. The effects of short-term incubation with these substances on the affinity of AChR were assessed by the rate of 125I-alpha-bungarotoxin-AChR complex formation. CMC desensitized AChRs, but MG globulins did not alter the affinity of AChRs. Results with this in vitro model suggest that chronic anticholinesterase therapy in the presence of MG AChR antibodies may aggravate failure of neuromuscular transmission in MG. PMID- 6277237 TI - Binaural voltage summation of brainstem auditory evoked potentials: an adjunct to the diagnostic criteria for multiple sclerosis. AB - Brainstem auditory evoked potential (BAEP) amplitude is modified according to whether or not the stimulus is applied monaurally or binaurally. In normal subjects, wave V amplitude increases by an average of 68.7% upon changing stimulation from monaural to binaural. From earlier studies there is evidence that brainstem potential amplitude is reduced in patients with multiple sclerosis (MS) but none to suggest that binaural stimulation results in increased amplitude. This study evaluated the extent of binaural summation of BAEPs in patients with MS. In a large majority of patients with MS who have no hearing deficit, BAEPs showed no increase in wave V amplitude on binaural stimulation. This finding is in contrast to the normal group and thus has diagnostic importance. Measurements of binaural summation therefore might usefully be applied to the clinical assessment of disease progression, or lack of it, in individual patients. PMID- 6277238 TI - Brain function, synapse renewal, and plasticity. PMID- 6277239 TI - Endorphins and behavior. PMID- 6277240 TI - Polymyxin antibiotics: chemical and pharmacokinetic properties. PMID- 6277241 TI - Pharmacokinetics of the new cephalosporins. PMID- 6277242 TI - [Effect of antibiotic and vaccine therapies on the succinate dehydrogenase and phosphatase activity of liver cells of mice infected with Staphylococcus]. AB - Changes in the activity of succinate dehydrogenase (SDH), total and acid phosphatase (TP and AP) were studied in treatment of laboratory animals with rifampicin, lincomycin and with inactivated staphylococcal vaccine used alone or in combinations. It was shown that immunization of the animals with inactivated staphylococcal vaccine under conditions of experimental staphylococcal infection promoted stimulation of the enzyme activity. Rifampicin and lincomycin used for the treatment of such animals lowered the activity of the enzymes. The suppressing effect of the antibiotics increased with an increase in the period of their use. It should be noted that the inhibitory effect of rifampicin on the activity of SDH, TP and AP was less pronounced than that of lincomycin. The combined use of the vaccine and antibiotics for the treatment of the animals promoted an increase in the enzyme activity as compared to the use of the antibiotics alone. Sometimes the activity of SDH, TP and AP reached the control levels in such animals or the levels observed in the animals treated with the vaccine alone. Stimulation of the enzyme activity was more pronounced when the vaccine was used in combination with rifampicin. PMID- 6277244 TI - Analgesia produced by stimulation of various sites in the human beta-endorphin system. PMID- 6277243 TI - [Modification of the poly G-poly C complex by incorporation of adenosine in the purine chain]. AB - Antiviral and interferonogenic activity of the complexes of poly(G,A) . poly(C) and poly(G) . poly(C) was studied in mice and cell cultures. Three out of 4 complexes of poly(G,A) . poly(C) had insignificant antiviral and interferonogenic activity in chick embryo cells. One of the complexes induced low levels of interferon production in mice and decreased the rate of their death from experimental forest-spring encephalitis. The activity of poly(G) . poly(C) in the above cell systems was much more pronounced. Unlike this complex, some complexes of poly(G,A) . poly(C) showed a noticeable activity in the cells of Primates. The effect of the noncomplementary base in the purine thread of poly(G) . poly(C) on its biological activity and nucleotide composition is discussed. PMID- 6277245 TI - Stimulation and coagulation of the posteromedial hypothalamus for intractable pain, with reference to beta-endorphins. PMID- 6277246 TI - Organization of basal ganglia inputs to the thalamus. A light and electron microscopic study in the cat. PMID- 6277247 TI - Chromosomal mapping of a gene affecting enterotoxin A production in Staphylococcus aureus. AB - In a previous study, transformation demonstrated that a gene governing enterotoxin A production (entA+) in Staphylococcus aureus strain S-6 was located on the chromosome between the purB110 and ilv-129 markers; in contrast, the entA+ gene of strain FRI-196E was shown not to be located in the same position. In the current study, 54 enterotoxin A-producing strains of S. aureus were examined to locate the entA+ gene. Conventional transformation procedures and a series of multiply marked derivatives of NCTC 8325 were used as recipients for chromosomal mapping. Of the 54 strains tested, 23 were found to contain the entA+ gene at the original locus between the purB110 and ilv-129 markers. Twenty-seven strains could not be analyzed either because their DNA was genetically ineffective in transforming strain 8325 (23 strains), or Pur+ Ilv+ transformants could not be recovered (four strains). Four other strains contained an entA+ gene that could not be located in any of the chromosomal linkage groups. A new insertion site for Tn551 was located within the hla+ gene involved in alpha-toxin production. It eliminated alpha-toxin production and was used to separate the entA+ gene from the hla+ marker in the purB110-ilv-129 region. This segment of the chromosome is shown to consist of the purB110, entA+, hla+, and ilv-129 markers in that order. PMID- 6277248 TI - Metal binding sites of rat liver Cu-thionein. PMID- 6277249 TI - Selective expression of type I and type II cyclic AMP-dependent protein kinases in subcellular fractions of concanavalin A-stimulated rat thymocytes. PMID- 6277250 TI - 1,24,25-Trihydroxyvitamin D3: a circulating metabolite in vitamin D2-treated bovine. PMID- 6277251 TI - Modulation of the 3',5'-cyclic AMP content by the respiration rate in E. coli K 12. PMID- 6277252 TI - Specific phosphorylation of eukaryotic initiation factor 2 alpha, eIF-2 alpha, by bovine adrenocortical cyclic nucleotide-independent protein kinase, PK 380. PMID- 6277253 TI - Purification and characterization of human lung calmodulin-independent cyclic AMP phosphodiesterase. PMID- 6277254 TI - Cytochrome changes in control and concanavalin A-stimulated lymphocytes. PMID- 6277255 TI - Hepatocellular carcinoma. PMID- 6277256 TI - Effects of nitric acid on carbachol reactivity of the airways in normal and allergic sheep. AB - The airway effect of a 4-hr exposure (via a Plexiglas hood) to 1.6 ppm nitric acid vapor were evaluated in seven normal and seven allergic sheep, i.e., animals that have a history of reacting with bronchospasm to inhalation challenge with Ascaris suum antigen. The nitric acid vapor was generated by ultrasonic nebulization of a 2% nitric acid solution. Airway effects were assessed by measuring the change in specific pulmonary flow resistance before and after a standard inhalation challenge with 2.5% carbachol aerosol. Nitric acid exposure did not produce bronchoconstriction in either group. Pre-exposure increases in specific pulmonary flow resistance after carbachol inhalation were 68% (SD +/- 13%) and 82% (SD +/- 35%) for the normal and allergic sheep, respectively. Within 24 hr, the largest post exposure increased in specific pulmonary flow resistance for the normal and allergic sheep were 108% [SD +/- 51% (P less than .06)] and 175% [SD +/- 87% (P less than .02)], respectively. We conclude that a short-term exposure to nitric acid vapor at levels below the industrial threshold limit (2 ppm), produces airway hyperreactivity to aerosolized carbachol in allergic sheep. PMID- 6277257 TI - [Heterologous Mullerian adenosarcoma of the uterus--a rare tumor. Light- and electron-microscopic findings, biological behavior (author's transl)]. AB - A case of heterologous Mullerian adenosarcoma of the uterus is presented with emphasis on morphologic differentiation, clinical course, prognosis, and treatment. This tumor was described first by Clement and Scully in 1974. Since that time 36 cases have been reported in the literature. The neoplasm consists of benign epithelial and malignant mesenchymal elements. The latter are composed of tissue with homologous and heterologous differentiation. Our microscopic and ultrastructural studies suggested that the heterologous elements were rhabdomyoblasts with a variable degree of differentiation. According to the literature the prognosis of Mullerian adenosarcoma is better than that of malignant mixed Mullerian tumor. The location of the tumor - whether intrauterine or extrauterine - seems to be more important as regards prognosis than the differentiation of the sarcomatous elements. The clinical course in our patients was not different from those described in the literature. Surgical removal of the tumor remains the treatment of choice. Some authors report a better survival rate when surgical treatment is followed by radiotherapy. Chemotherapy may be useful in cases of local or distant metastasis, and in cases of incomplete excision of the tumor mass. PMID- 6277258 TI - Nipple discharge from the breast. AB - Abnormal nipple discharge is rare, constituting only 3-5% of mammary consultation. In the present paper 267 patients with primary nipple discharge operated on at Institut Gustave-Roussay (IGR) in Villejuif, France, between January 1, 1960 and December, 1974 were evaluated. In all cases, the nipple discharge was symptomatic, spontaneous, and represented the primary reason for the patient's consultation. During the same period 1,145 cases of nipple discharge were treated at IGR. Of the 1,145 cases with symptomatic nipple discharge, 267 patients (23%) required surgical intervention. Among these, fibrocystic disease and duct ectasia were the leading causes of nipple discharge occurring in 42% of surgical specimens. Twenty one per cent of the patients had carcinoma and 35% were found to have intraductal papilloma. The overall incidence of malignancy, however, was 4.8% among the 1,145 women with nipple discharge. On the average, patients with nipple discharge due to malignancy were ten years older than those with benign lesions (Table 2). Approximately 25% of patients with malignant discharge and 5% with benign discharge have associated tumor. Over 60% of the patients with both discharge and a mass had malignancy. PMID- 6277259 TI - Surgical experience with pancreatic and periampullary cancer. AB - Between 1940 and 1978, 179 patients underwent pancreatic resection (64 total, 102 Whipple, 13 distal) at the Presbyterian Hospital, predominantly for carcinoma of the pancreas and periampullary area. With respect to operative morbidity and mortality and survival, these patients have been compared with 141 patients subjected to pancreatic biopsy only, and with 172 by-passed for palliation. Likewise, total pancreatectomy has been compared to pancreaticoduodenectomy (Whipple) in terms of safety and efficacy. The overall major postoperative complication rate for pancreatic resection was 36%, in contrast with 13.5% for biopsy only and 16.8% for by-pass. Of the resected cases with major complications postoperatively, roughly half died, a mortality of 17.9%. Patients who underwent Whipple resections fared significantly better than did those having total pancreatectomies; the postoperative mortality following 102 Whipples was 14.7%, as compared with 23.4% for total pancreatectomies. Intra-abdominal sepsis accounted for most of the postoperative deaths; nine pancreatic and four biliary leaks or fistulae followed Whipple resections. The later complications were of interest; 18 patients undergoing biliary-en-teric by-pass procedures later developed gastroduodenal obstruction, 15 of whom required reoperation, and in 18 survivors of pancreatic resection, upper gastrointestinal hemorrhage (mostly from marginal ulcers) developed, necessitating surgery in seven. Brittle diabetes was a problem in nine patients following pancreatectomy. Survival rates were discouraging in all categories. For ductal carcinoma of the pancreas, median survival for biopsy only was two months, for by-pass six months, for total pancreatectomy nine months, and for Whipple resection 14 months. There were three five-year survivors following resection, a rate of 4.5%. Five-year survival rates following resection for ampullary, common duct, duodenal, and islet cell cancer were 27.8, 33.3, 27.3, and 37.5%, respectively. It is concluded that survival after resection for ductal pancreatic cancer is so rare as to be considered more a biologic aberration than a result of radical surgery. Despite theoretical advantages of total pancreatectomy over Whipple resections, our experience would suggest that the latter can be carried out with lower morbidity and mortality, and with equal chance for cure. Resection for pancreatic cancer should not be abandoned, but rather undertaken with greater selectivity. Operative morbidity and mortality can probably be improved additionally by preoperative transhepatic biliary decompression, and later complications reduced by including vagotomy with gastric resection at the time of pancreatectomy and by performing prophylactic gastroenterostomies in conjunction with by-pass procedures. PMID- 6277260 TI - Immune responses to varicella-zoster in the aged. AB - Skin test reactivity and in vitro lymphocyte stimulation responses to varicella zoster (VZ) were examined in a large normal population ranging in age from 6 months to 93 years. Warning of cellular immunity, as examined by skin delayed hypersensitivity, began at age 40 years. Skin test responses to phytohemagglutinin, however, remained positive into the eighth decade of life. In vitro lymphocyte stimulation responses to VZ were usually positive (stimulation index greater than or equal to 2.5) until age 60 years, after which time levels, as observed with nonimmune individuals, were often demonstrated. Antibody levels, as measured by fluorescent antibody to membrane antigen, remained positive into the ninth and tenth decades of life. This was especially so with a history of reactivation (zoster) VZ infections, while skin test and in vitro responses were rarely positive in those individuals. This cellular, as contrasted with humoral, immunity decreases with advancing age, which may account for a propensity to reactivation of VZ virus. PMID- 6277261 TI - Rotavirus infection in a geriatric population. AB - An outbreak of gastroenteritis affected 19 of 34 geriatric patients and four of 23 staff assigned to the ward in a period of 3 1/2 weeks in January 1980. Fourteen of the 19 patients with gastroenteritis (17 were tested properly) and four of the ten asymptomatic patients (five asymptomatic patients were not tested) showed evidence of rotavirus infection by virus positivity and/or a significant antibody response to rotavirus. One of the four staff members with gastroenteritis showed serologic evidence (three were tested) of rotavirus infection. Nine of the 18 asymptomatic staff members (two remaining staff members were not tested) showed a fourfold rise in antibody to rotavirus but four had antibody titers of 1:32 or more. The patients had diarrhea for a mean of 2.6 days. Most of them had five or fewer diarrheal stools in one day. Six patients had a severe illness and two died. Thirteen of 15 symptomatic patients who had serum samples, collected during the acute and convalescent phases, tested manifested high titers (greater than or equal to 1:32) of complement-fixing antibody to rotavirus antigen. PMID- 6277262 TI - Small cell anaplastic carcinoma of the lung with mesangial proliferative glomerulonephritis. AB - Mesangial proliferative glomerulonephritis is an uncommon manifestation of renal injury associated with neoplastic disease. A 50-year-old woman with small cell anaplastic cancer of the lung and nephrotic syndrome had renal biopsy findings that were consistent with diffuse mesangial cell proliferation. Electron microscopic evaluation of renal tissue demonstrated numerous intramesangial and paramesangial dense deposits. Resolution of the nephrotic syndrome with improvement in renal function was noted after a response of the patient's tumor to combination chemotherapy. PMID- 6277263 TI - Large single doses of oral TRH blockade the TSH response in children. PMID- 6277264 TI - Rapid diagnosis of amoebic liver abscess using Entamoeba histolytica antigen. PMID- 6277265 TI - [Comparison of the global and local contractility of the left ventricle by volumetric, radioisotopic and angiographic methods]. PMID- 6277266 TI - Regulation of exoprotease production by temperature and oxygen in Vibrio alginolyticus. AB - The production of an extracellular collagenase and alkaline protease by Vibrio alginolyticus during stationary phase was inhibited by a temperature shift from 30 to 37 degrees C and by a lack of oxygen. The stability of the exoproteases was unaffected by incubation at 37 degrees C and aeration. The optimum growth temperature for the V. alginolyticus strain was 33.5 degrees C and there was no difference in the growth rate at 30 and 37 degrees C. Aeration enhanced the rate of growth of exponential phase cells. Temperature and oxygen did not affect the growth of stationary phase cells when the exoproteases were being produced. Macromolecular synthesis in stationary phase cells was not affected by temperature. There was no rapid release of the exoproteases after temperature shift down and chloramphenicol inhibited the production of the enzymes when added at time of temperature shift down from 37 to 30 degrees C. The regulation of exoprotease production by temperature and oxygen was specific and has implications regarding the ecology of V. alginolyticus. Cerulenin, quinacrine and O-phenanthroline inhibited the production of the exoproteases. PMID- 6277267 TI - Use of an electroimmunodiffusion method on cellulose acetate films for the detection of antigenicity in human breast cancers that is related to the major core antigen of mouse mammary tumor virus (MuMTV). AB - A highly sensitive electroimmunodiffusion method on cellulose acetate films, developed by Abelev on the basis of isotachoelectrophoresis, was applied to search of MuMTV-related antigenicity in tumor tissue and sera of breast cancer patients. In 2 out of 5 breast cancer extracts, protein fractions corresponding to a molecular weight of about 50,000 daltons were seen to exhibit antigenicity related to the major core antigen of MuMTV (p27). This antigenicity could not be detected in the sera of the patients. PMID- 6277268 TI - [Carcinoembryonic antigen in serum and tissue of benign and cancerous breast lesions]. AB - The tissular CEA was studied with the immunoperoxidase method and the circulating CEA with the enzyme immunoassay (EIA), using peroxidase-coupled antibodies. 51 cases of mammary cancers as well as 33 benign lesions were examined in order to study a possible correlation between the frequency of CEA-positive tissues and CEA-positive sera in the same patient. It has been confirmed that CEA-positive tumors can be accompanied by low or negative CEA serum levels. An attempt was made to analyze the stage and the histologic type of lesions in comparison to the positivity and the increase of CEA. PMID- 6277269 TI - Functional changes of macrophages from polyoma tumor-bearing mice. AB - CBA mice were inoculated with different doses of polyoma tumor cells. The PHA response of spleen cells from tumor-bearers was decreased but the spontaneous incorporation of 3H-leucine was increased. PHA-induced cytotoxicity of spleen cells was increased only in tumor-bearers. The expression of Fc receptors showed a slight increase on adherent cells from spleen and peripheral blood mononuclear cells under suboptimal conditions for EA rosette formation (small amount of antibody for coating the erythrocytes). The non-specific esterase activity was assayed in an extract of peritoneal macrophages and the elevated activity with isoenzymes was seen in a group of tumor-bearers. The results suggest a functional alteration of macrophages in polyoma tumor-bearing mice. PMID- 6277270 TI - Montgomery's areolar tubercle. A light microscopic study. AB - Conflicting impressions regarding the anatomy of Montgomery's areolar tubercle exist. Twelve modified radical mastectomy specimens provided 1,536 serial sections of areolar tubercles. In 34 of 35 tubercles (97%), a mammary lactiferous duct was associated with a sebaceous apparatus. This lactiferous duct ascended from deeper mammary parenchyma and entered the sebaceous gland. Histopathologic changes identified included featured of fibrocystic disease, atypical intraductal hyperplasia, and carcinoma in situ. Because the areolar tubercle has two components, a sebaceous gland and a mammary duct arising from deeper breast parenchyma, diseases of the breast may also involve the areola independent of papilla-nipple involvement. Areolar preservation may best be used with the knowledge that diseases underlying the areola may also involve the areola. PMID- 6277271 TI - An immunoperoxidase study of cytomegalovirus mononucleosis. AB - Indirect immunoperoxidase method was used to evaluate retrospectively a case of cytomegalovirus (CMV) mononucleosis. Specific rabbit antiserum against CMV was used as primary reagent in routinely prepared paraffin sections of lymph node and in fixed slide preparations of the bone marrow aspirate. Staining specific for CMV antigen was observed in the lymphoid cells of the lymph node in the predominantly T-cell area and in the large lymphoid cells of the bone marrow. Bone marrow lymphoid cells with morphologic characteristics similar to those of the cells positive for CMV antigen showed positive immunoperoxidase reaction with specific rabbit antihuman T-cell serum. The data strongly indicate that CMV antigen was localized predominantly in the cells of lymphoid origin during the acute stage of the CMV mononucleosis; some data also suggest that infected lymphoid cells may be of T-cell origin. An immunoperoxidase technique is advantageous in both diagnosing and evaluating CMV mononucleosis. PMID- 6277272 TI - Monophasic, glandular, synovial sarcomas and carcinomas of the soft tissues. AB - In soft tissues, malignant tumors with epithelial qualities are usually metastatic carcinomas. A rare, primary, soft-tissue tumor that is uniformly carcinoma-like is the monophasic, glandular, synovial sarcoma. We studied three cases of carcinoma-like neoplasms of soft tissue. In one primary tumor and two recurrent tumors of soft tissue, the histologic patterns were uniformly glandular. For the two recurrent tumors, the primary lesions were a classic, synovial sarcoma and a biphasic, synovial sarcoma with a histiocytic, stromal component. By inference, the third tumor, a primary "carcinoma" of soft tissue, may represent a de novo, monophasic, glandular, synovial sarcoma. Synovial sarcoma should be considered in the differential diagnosis of carcinoma like tumors discovered in the soft tissues. PMID- 6277273 TI - Antipertussis immune response after 2 and after 3 doses respectively of diphtheria-tetanus-pertussis trivaccine. PMID- 6277274 TI - [Computerized axial tomography in the study of primary bone tumors of the extremities]. PMID- 6277275 TI - [ulnar nerve neuropathy at the epitrochlear sulcus]. PMID- 6277276 TI - [experimental reproduction of synovial sarcoma]. PMID- 6277277 TI - [Non-osteogenic fibroma of the long bones. Clinico-pathological and therapeutic considerations on a review of 18 cases. Theory of possible identification in benign fibrous histiocytoma]. PMID- 6277278 TI - Diabetic sexual dysfunction: a comparative study of 160 insulin treated diabetic men and women and an age-matched control group. PMID- 6277279 TI - Effect of a single oral dose of pp'DDT on the absorption of nutrients in vitro and on brush border enzymes in rat intestine. AB - The effect of a single oral dose of pp'DDT (100 mg/kg body wt.) has been studied on the intestinal uptake of certain nutrients and on brush border enzymes in rats. Intestinal uptake of leucine, and phenylalanine was considerably increased but there was no change in the absorption of glucose and alanine in DDT fed rats, compared to controls. The activities of brush border sucrase, alkaline phosphatase and Na+, K+-ATPase were significantly depressed in pesticide treated animals, but leucine aminopeptidase levels remained unaffected under these conditions. Analysis of the chemical composition of the microvillus membranes revealed a considerable enhancement in total lipids, phospholipids and triglyceride contents of the membranes in DDT exposed rats, but membrane protein, sialic acid and cholesterol fractions did not record any change. 1-14C-acetate incorporation into various lipid classes was studied to explain the observed increase in membrane lipids in DDT exposed animals. PMID- 6277280 TI - The intracellular proteins induced by cricket paralysis virus in Drosophila cells: the effect of protease inhibitors and amino acid analogues. AB - Treatment of Cricket paralysis virus infected Drosophila cells with iodoacetamide before radiolabelling with 35S-methionine results in the appearance of two high molecular weight polypeptides of approximately equal to 200,000 molecular weight, not apparent in untreated infected cells (17). To attempt to differentiate between the effects of iodoacetamide being attributable to either alteration of initial polyprotein or inhibition of the protease (either cellular or viral) the effects of a spectrum of protease inhibitors were examined. These included aprotinin, leupeptin, pepstatin, elevated zinc concentration, phenyl methyl sulphonyl-fluoride, N-tosyl-L-lysine chloromethyl ketone (TLCK) and N-tosyl-L phenylalanine chloromethyl ketone (TPCK). TLCK and TPCK both inhibited the cleavage of proteins which demonstrates an inhibition of the protease activity. The introduction of amino acid analogues into the infected cells before pulsing also results in the appearance of higher molecular weight proteins. This could be attributed to alternation of the polyprotein making it nonsusceptible to digestion with pre-existing cellular protease or newly synthesized viral protease. The possibility that the presence of the amino acid analogues results in alteration of a viral coded protease cannot be eliminated. PMID- 6277281 TI - Thermal stability of foot-and-mouth disease virus. AB - The thermal stabilities of 146S component of seven strains of foot-and-mouth disease virus were found to differ considerably. Inactivation of infectivity with acetylethyleneimine (AEI) reduced the thermal stabilities of all but one of the viruses. Treatment of AEI inactivated and control virus preparations with glutaraldehyde stabilized 146S particles to a considerable extent, whereas treatment with dimethyl suberimidate was less effective. In similar experiments with 75S, natural empty particles, the thermal stabilities were lower than those of the corresponding 146 S particles. Treatment of 75S particles with AEI appeared to have no direct effect on the protein-protein interactions involved in 75S capsid integrity. As with 146S particles, glutaraldehyde stabilized 75S particles. PMID- 6277282 TI - Morphogenesis of human rotavirus type 2 Wa strain in MA 104 cells. AB - Morphogenesis of human rotavirus type 2 Wa strain in MA 104 cells was observed. The virus antigen in the cytoplasm was detected by indirect immunofluorescence twelve hours after infection. The cytopathic effect occurred 24 hours after infection when virus particles were detected by EM in the culture fluid as well as in thin sections of the infected cells. Virus particles were observed in the dilated RER, nuclear envelope (perinuclear space), viroplasm, and a lysosome-like body. Three types of virus particles were noted: double-shelled particles 75-85 nm in diameter, single-shelled particles 64-68 nm in diameter and electron-dense nucleoids or cores 32-40 nm in diameter. The outer shell of virus particles was acquired by budding through the membrane of the dilated RER. Tubular structures, similar in diameter to the single-shelled particles, were found in the cytoplasm and nucleus of infected MA 104 cells. Bundles of filaments or the leaflet-like inclusion bodies of membrane-bounded bundles of filaments were found in the cytoplasm and seemed to be associated with virus particles. PMID- 6277283 TI - Relationship between plaque size and the immunising ability of the foot-and-mouth disease virus SAT 1 Nig 10/75. PMID- 6277284 TI - Comparison of mouse hepatitis virus strains for pathogenicity in weanling mice infected by various routes. PMID- 6277285 TI - Salt-dependent hemagglutination with bluetongue virus. PMID- 6277286 TI - Diagnosis of Junin virus in cell cultures by immunoperoxidase staining. PMID- 6277287 TI - Argentine hemorrhagic fever: early diagnosis by immunofluorescence. AB - A procedure that allows to ascertain a diagnosis of Argentine Hemorrhagic Fever (AHF) as early as 24-48 hours after onset of symptoms is described. An immunofluorescent (IF) test on round cells of urinary sediment was employed. The procedure was assayed on 31 patients with febrile syndrome during epidemic peaks of 1975-1976. It was positive in 19 and negative in 12 cases. The 19 positive cases were confirmed AHF by clinical follow up and serology. From 12 negative cases, 8 belonged to other etiologies and 2 were confirmed AHF. The usefulness of the procedure for early diagnosis is emphasized. PMID- 6277288 TI - Determination of immunity to varicella using an enzyme-linked-immunosorbent assay. PMID- 6277289 TI - A review of the antiemetic use of delta-9-tetrahydrocannabinol and prescribing requirements. PMID- 6277290 TI - Lipids associated with mineralization of human submandibular gland sialoliths. PMID- 6277291 TI - Clinical improvement and hormonal changes in severe cardiac failure after captopril treatment. PMID- 6277292 TI - Schooling behavior of fish: an opiate-dependent activity? PMID- 6277293 TI - Loss of canine myocardial nicotinamide adenine dinucleotides determines the transition from reversible to irreversible ischemic damage of myocardial cells. AB - We investigated if the loss of nicotinamide coenzymes in ischemic-infarcted myocardium may be responsible for the transition from reversibly ischemic to irreversibly infarcted cell damage. The LAD was occluded in 6 dogs for 4 h. Transmural needle biopsies were taken from he ischemic-infarcted region after 1/2, 1, 11/2, 2, and 4 h of ischemia and further divided into subepicardial and subendocardial halves. At each time interval the concentration of the nicotinamide coenzymes NAD, NADH, and NADPH were measured, and the degree of cellular injury was evaluated by electron microscopy. The glycohydrolase activity (EC 3.2.2.5), the enzyme which splits NAD, was determined in brain, myocardium, kidney, and skeletal muscle of 4 rats. Total NAD, the sum of NAD and NADH, started to decrease significantly in the ischemic subendocardium 1 h after onset of ischemia. Degradation of NADPH occurred later. Loss ot total NAD was about 60 70% when electron microscopy diagnosed irreversible cell injury. The glycohydrolase activity was the highest in brain followed by myocardium, kidney, and skeletal muscle, reflecting the different tolerances of these tissues towards ischemia. The key mechanism for ischemic injury seems to be the tissue acidosis which activates the glycohydrolase leading to a loss of the vital coenzymes. PMID- 6277294 TI - Enrichment of vital adult cardiac muscle cells by continuous silica sol gradient centrifugation. AB - A major improvement in the isolation of vital adult cardiocytes was achieved by isopycnic preformed continuous silica sol gradient centrifugation after perfusion of the heart with collagenase. Vital rat cardiocytes were enriched to 90-95% vital cells reproducibly and constantly by one- or two-step gradient centrifugations. The isolated cardiocytes were tolerant to calcium concentrations up to 0.03 mmol/l, to diluted human serum, and to human complement. Gentamycin (50 microgram/ml) exerted a cytotoxic effect on myocytes, whereas Penicillium and Streptomycin in concentrations of 50 IU/ml did not induce cytolysis of vital cells. Digoxin 15 ng/ml) decreased the natural decay of myocytes of 20% in 25 hours to 8%. Enriched of vital cardiocytes by silica sol gradient centrifugation following their isolation by perfusion with collagenase may be helpful for investigations depending on a high yield of vital myocardial cells. PMID- 6277295 TI - [Diagnosis of canine herpes virus infection (CHV)]. PMID- 6277296 TI - Receptor-mediated gonadotropin action in the ovary. Inhibitory actions of concanavalin A and wheat-germ agglutinin on gonadotropin-stimulated cyclic AMP and progesterone responses in ovarian cells. AB - Pretreatment of ovarian cells with concanavalin A and wheat-germ agglutinin blocked the gonadotropin-induced cyclic AMP and progesterone responses and this effect was time- and concentration-dependent. Basal production of either cyclic AMP or progesterone, however, was not affected by treatment of cells with lectin. The effect of concanavalin A on gonadotropin-mediated cyclic AMP and progesterone responses was blocked by alpha-methyl D-mannoside and alpha-methyl d-glucoside. Similarly the inhibitory effect of wheat-germ agglutinin was reversed by N-acetyl D-glucosamine. Pretreatment of ovarian cells with concanavalin A or wheat-germ agglutinin had no effect on protein synthesis in the ovary as monitored by [3H]proline incorporation studies. Concanavalin A and wheat-germ agglutinin did not affect steroid production in response to dibutyryl cyclic AMP and 8-bromo cyclic AMP, indicating that the inhibitory action of lectin was occurring at a step before cyclic AMP formation. Lectins specific for L-fucose, D-galactose and N-acetyl-D-galactosamine, gorse seed agglutinin, peanut agglutinin and Dolichos biflorus agglutinin respectively, did not interfere with gonadotropin-induced cyclic AMP and progesterone responses. The present studies suggest that gonadotropin receptors may be glycoprotein in nature or closely associated with glycoprotein structures with the carbohydrate chain containing N-acetyl-D glucosamine, mannose and possibly N-acetylneuraminic acid. PMID- 6277297 TI - Combined radiolabel-binding and immunocytochemical evaluation of receptor-ligand interactions. Studies of transferrin receptors on activated lymphocytes. AB - A protocol that involved both immunohistological and radiolabel-binding procedures was devised for the study of transferrin-receptor interactions. This composite approach yielded considerably more information than did either technique used alone, and also provided a simple means for exclusion of several common potential sources of error. PMID- 6277298 TI - Receptor-mediated vitellogenin binding to chicken oocytes. AB - The specific binding of vitellogenin to chicken oocyte membranes was characterized. This major hen serum phospholipoglycoprotein and one of its lower molecular-weight components, phosvitin, bound to oocyte membranes with KD values of approx. 6 x 10-7 M. The optimum pH for binding was 6.0, the same as the pH of yolk contents. Phosvitin and vitellogenin compete with each other for binding; other proteins tested do not compete to the same degree. Phosvitin, which contains 10% phosphate by weight, appears to be the polypeptide recognized by the receptor. RNA failed to compete with either vitellogenin or phosvitin for binding, suggesting that the binding specificity may require more than polymeric phosphate. The binding was tissue-specific in that phosvitin and vitellogenin bound to oocyte surfaces (at both pH 6.0 and 7.5), but not to chicken erythrocytes (at either pH). PMID- 6277299 TI - The influence of various proteases and inhibitors on steroid hormone receptors in rat liver and kidney. PMID- 6277301 TI - (Na+ + K+)-ATPase activity of newborn rabbit cardiac muscle. PMID- 6277300 TI - Altered kinetics of membrane ATPase in mental illness. PMID- 6277302 TI - Acetoacetate is an electron donor to myeloperoxidase and a promoter of myeloperoxidase-catalyzed fatty acid peroxidation. PMID- 6277303 TI - Influence of hexose conditions on glutamine oxidation of SV-40-transformed and diploid fibroblast human cell lines. PMID- 6277304 TI - Levels of adenosine deaminase AMP deaminase, and adenylate kinase in cultured human lymphoblast lines: exquisite sensitivity of AMP deaminase to adenosine deaminase inhibitors. PMID- 6277305 TI - Altered isoelectric property of a superactive 5-phosphoribosyl-1-pyrophosphate (PRPP) synthetase in a patient with clinical gout. PMID- 6277306 TI - Similarities in the molecular weight of poly(ADPR) polymerase from different tissues and species. PMID- 6277307 TI - Uncoupling of acetylcholine uptake from the Torpedo cholinergic synaptic vesicle ATPase. PMID- 6277308 TI - Ribonuclease F, a putative processing endoribonuclease from Escherichia coli. PMID- 6277309 TI - Evidence for 5, 12-dihydroxy-6,8,10,14-eicosatetraenoate as a mediator of human neutrophil aggregation. PMID- 6277310 TI - Nucleotide sequence of the genes coding for alpha, beta and gamma subunits of the proton-translocating ATPase of Escherichia coli. PMID- 6277311 TI - Nucleotide sequence of the genes for F0 components of the proton-translocating ATPase from Escherichia coli: prediction of the primary structure of F0 subunits. PMID- 6277312 TI - Prevention of prostaglandin E2-induced desensitization of rat ovarian cyclic AMP response by concanavalin A. PMID- 6277313 TI - Influence of isoproterenol on net potassium uptake in whole pigeon erythrocytes in vitro. PMID- 6277314 TI - An estrogen-stimulated 1,25-dihydroxyvitamin D3 receptor in rat uterus. PMID- 6277315 TI - Arachidonic acid, bradykinin and phospholipase A2 modify both prolactin binding capacity and fluidity of mouse hepatic membranes. PMID- 6277316 TI - Rapid changes in the activities of the enzymes of cyclic AMP metabolism after addition of A23187 to macrophages. PMID- 6277317 TI - Deactivation of the effects of F-Met-Leu-Phe and leukotriene B4 on calcium mobilization in rabbit neutrophils. PMID- 6277318 TI - Covalent binding of benzo[a]pyrene to dna in fish liver. PMID- 6277319 TI - Evidence for protein phosphorylation as a regulatory mechanism for hepatic microsomal glucose 6 phosphatase. PMID- 6277320 TI - The effect of PEA on serotonin and catecholamines. PMID- 6277321 TI - Interaction of vasoactive intestinal peptide with HeLa cells: activation of cyclic AMP-dependent protein kinase and lack of effect on DNA synthesis. PMID- 6277322 TI - Cloning and sequencing of cDNA for mouse liver metallothionein-I. PMID- 6277323 TI - D-Fructose 2,6-bisphosphate: a naturally occurring activator for inorganic pyrophosphate:D-fructose-6-phosphate 1-phosphotransferase in plants. PMID- 6277324 TI - Regulation of fructose-1,6-bisphosphatase in yeast by phosphorylation/dephosphorylation. PMID- 6277325 TI - Structure and conformation of the potent antiherpes agent 9-(2 hydroxyethoxymethyl) guanine (acycloguanosine). PMID- 6277326 TI - Acetylated and nonacetylated forms of beta-endorphin in rat brain and pituitary. PMID- 6277327 TI - Stimulating factor for calf thymus DNA polymerase beta. PMID- 6277329 TI - Free radical metabolism of mutagenic acridines and binding to microsomal membranes. PMID- 6277328 TI - The submitochondrial localization adenylate kinase: an enzymatic marker for the inner surface of the outer membrane of lung mitochondria in guinea pig. PMID- 6277330 TI - Identification of cysteines in subunit II as ligands to the redox centers of bovine cytochrome c oxidase. PMID- 6277331 TI - Role of polyamines in the proliferation and steroidogenic activities of bovine adrenocortical cells in culture. PMID- 6277332 TI - Cyclic AMP-binding protein and estrogen receptor: antagonism during nuclear translocation in a hormone-dependent mammary tumor. PMID- 6277333 TI - The role of possible membrane damage in porphyria cutanea tarda: a spin label study of rat liver cell membranes. AB - Two groups of rats were made porphyric by treatment for a shorter or a longer time with hexachlorobenzene (HCB); a third group was subjected to chronic treatment with ethanol; the fourth group comprised untreated, control animals. Suspensions were prepared of hepatocytes isolated from the livers of the animals in the individual groups, and in these systems the cell membranes were studied by spin labeling. In all three chronically treated groups, the order parameter calculated from the ESR spectra was significantly lower than that of the control group. The order parameters ofr the first three treated groups, however, did not differ significantly from one another. If either HCB or ethanol was incorporated into the isolated hepatocytes of the healthy control animals, the fluidity of the hepatocyte membranes increased. The order parameter decrease (compared to the control) for the hepatocyte membranes of the animals treated chronically with HCB or ethanol can be ascribed to the direct membrane-fluidizing effect of HCB or ethanol, and also to the altered lipid metabolism. With regard to the difference in the mechanisms of action of HCB and ethanol, on the basis of the experimentally proved membrane-damaging effect of the porphyrinogenic HCB it is probable that, besides other factors, the membrane damage may play an important role in the pathogenesis of porphyria cutanea tarda. PMID- 6277334 TI - In vitro desensitization of human lymphocytes by epinephrine. PMID- 6277335 TI - Different numbers of beta-receptors in human lymphocyte subpopulations. PMID- 6277336 TI - Differential effects of various phosphodiesterase inhibitors, pyrimidine and purine compounds, and inorganic phosphates on cyclic CMP, cyclic AMP and cyclic GMP phosphodiesterases. AB - The effects of various compounds on homogeneous cyclic CMP phosphodiesterase (cyclic CMP-PDE) from pig liver were compared with the effects on cyclic AMP phosphodiesterase (cyclic AMP-PDE) and cyclic GMP phosphodiesterase (cyclic GMP PDE). Of the conventional inhibitors for AMP-PDE and cyclic GMP-PDE, only Sch 15280 was found to inhibit cyclic CMP-PDE. Nucleoside monophosphates, orthophosphate, and 2':3'-cyclic nucleotides were rather specific and were more effective in inhibiting cyclic CMP-PDE, compared to their effects on cyclic AMP PDE and cyclic GMP-PDE. On the other hand, nucleoside di-and triphosphates and pyrophosphate (PPi) were less effective in inhibiting cyclic CMP-PDE and were without marked effect on cyclic AMP-PDE and cyclic GMP-PDE. Orthophosphate (Pi) was more potent than CMP, CDP and CTP in inhibiting cyclic CMP-PDE, with a rank order of inhibitory potency of Pi greater than CMP greater than CDP greater than CTP. Of the 3' :5'-cyclic nucleotides examined, cyclic UMP was more specific in inhibiting cyclic CMP-PDE compared to its effect on cyclic AMP-PDE and cyclic GMP PDE. In all experiments similar results were obtained when either cyclic CMP or cyclic AMP was used as a substrate for this multifunctional cyclic CMP-PDE, supporting the contention that a single catalytic site on the enzyme is responsible for the hydrolysis of both cyclic CMP and cyclic AMP. The present studies further support our original suggestion that cyclic CMP-PDE is a unique enzyme that is distinguishable from the conventional enzymes for purine cyclic nucleotides. PMID- 6277337 TI - Relationship of brain cyclic nucleotide levels and the interaction of ethanol with chlordiazepoxide. AB - The effects of combined administration of ethanol (4 g/kg) and chlordiazepoxide (CDP, 12.5 mg/kg) on mouse brain c-AMP and c-GMP levels were investigated in order to test the hypothesis that the supra-additive effect of CDP on ethanol sleep time may be related to a supra-additive alteration in brain cyclic nucleotide levels induced by the combined drugs. Ethanol alone or CDP by itself did not cause any change in brain c-AMP levels, except for a transient decrease in the cerebral cortex and midbrain at 0.5 hr after ethanol injection, as well as a transient increase in the cerebellum at 0.5 hr after CDP injection. The combined drug treatment did not result in a supra-additive effect on c-AMP levels. On the other hand, c-GMP levels were depressed significantly for 4 hr after ethanol injection especially in the cerebellum. The mice regained the righting reflex when the c-GMP levels were still about 30 per cent of control values. Ethanol and CDP together induced a supra-additive decrease of c-GMP concentrations in the cerebellum at 2 and 4 hr. This resulted in a lengthened period (about 2.5 hr) during which the cerebellar c-GMP levels were below 30 per cent of control values, and this interval coincided with the increase in sleep time, suggesting a possible relationship between these two factors. Injection of ethanol and N-demethyl-chlordiazepoxide (NDCDP) simultaneously (the latter being a metabolite of CDP) also elicited a more than additive depression of cerebellar c-GMP levels at 4 hr. These data suggest that NDCDP or its metabolite could be responsible for the supra-additive effect of CDP on the ethanol-induced decrease in cerebellar c-GMP levels. PMID- 6277338 TI - Receptor binding sites for beta-adrenergic ligands on human erythrocytes. AB - Affinity, specificity and kinetics for [3H]-DHA binding to human red cell ghost were determined by ultra-filtration. At 2 degree an apparent dissociation constant of 0.96 nM was found with maximum specific binding of 29 fmoles per mg protein. The low dissociation constant was confirmed by kinetic studies with a value of 0.86 nM. Propranolol and isoproterenol inhibited [3H]-DHA binding stereo specifically. Agonist potency (IPR greater than EPI greater than NE) indicated that human erythrocytes had an adrenergic receptor of beta-2 subtype. Isoproterenol in the presence of theophylline resulted in a concentration dependent increase of intracellular cAMP levels in intact cells. Basal and maximal levels were 2.3 and 7.5 pmoles/108 cells respectively after 2.5 min stimulation. EC50 for isoproterenol was 0.27 microM. Propranolol shifted the isoproterenol concentration response curve to the right. The present results show that human erythrocytes possess recognition sites for beta-adrenergic ligands with binding characteristics similar to that of adrenergic receptors of beta-2 subtype. At least a small number of these binding sites are functionally coupled to adenylate cyclase. PMID- 6277339 TI - Cell cycle specific fluctuations of adenosine 3',5' -monophosphate and prostaglandin binding in synchronized mastocytoma P-815 cells. AB - Endogenous adenosine 3',5' -monophosphate (cAMP) levels in mastocytoma P-815 cells, synchronized either at the G1/S transition by amethopterin- or double thymidine-block or in mitosis by colcemid block, were highest during late S and early G2 phases and lowest during mitosis. These cell cycle-dependent changes in cAMP levels were largely accounted for by changes in adenylate cyclase and phosphodiesterase activities. Similar fluctuations occurred simultaneously with specific prostaglandin E1 (PGE1) binding, histidine decarboxylase activity, histamine content, and [35S]SO-2(4) incorporation into glycosaminoglycans of the cells. In addition, endogenous levels of the E group of prostaglandins (PGEs) and "14C]carachiodonic acid incorporations into PGE, phosphatidylcholine and phosphatidylinositol also exhibited fluctuation patterns similar to that of cAMP levels. Since cAMP levels still fluctuated in a serum-depleted medium where DNA synthesis and cell division were inhibited, endogeneous levels of prostaglandin and cAMP appeared not to be regulated solely by serum factor(s). Exposure of cells at G1/S transition to 1-methyl-3-isobutylxanthine (MIX) resulted in 10-fold elevation of cAMP levels throughout the cell cycle without affecting DNA synthesis. On the other hand, PGE1 and/or MIX added at late S phase elevated cAMP levels, prolonged C2 phase and retarded the cell division, but these agents added at the beginning of mitosis elevated cAMP levels without affecting the cell division. These results suggest that prostaglandin newly synthesized by the increased metabolism of phospholipids promote the cAMP synthesis via their binding to the receptors and thereby control the division and phenotypic expression of mastocytoma P-815 cells. PMID- 6277341 TI - Water versus acetone-HCl extraction of digitalis-like factor from guinea-pig heart. PMID- 6277340 TI - Mixed function oxidase in the mammary gland and liver microsomes of lactating rats. Effects of 3-methylcholanthrene and beta-naphthoflavone. AB - Mammary gland and liver microsomes of lactating rats were examined for the components of mixed function oxidase and related enzyme activities. Cytochrome b5, NADH- and NADPH- dependent cytochrome c reductase activities were 15-, 6- and 10-fold lower, respectively, in the mammary gland than in the liver microsomes. The determination of cytochrome P-450 (P-448) in the mammary gland microsomes required elimination of the spectral interferences by hemoglobin and cytochrome aa3. The presence of the latter in this fraction was also shown by cytochrome c oxidase activity. Cytochrome aa3 was reduced by anaerobic incubation of mammary gland microsomes, in the presence of antimycin A, with sodium succinate, phenazine ethosulfate, and sodium ascorbate for 30 min at room temperature. Spectral resolution of the dithionite-reduced cytochrome P-450 (P-488) carbon monoxide complex occurred 30 min after gassing. The basal level of cytochrome P 450 was about 500-fold greater in the liver than in the mammary gland microsomes. Pretreatment of lactating rats with the inducers of hepatic cytochrome P-448, 3 methylcholanthrene and beta-naphthoflavone, increased the cytochrome content 3- to 10-fold, in the mammary gland and liver microsomes, respectively. The induction of cytochrome P-448 in microsomes of both tissues was also shown by type I binding spectra obtained with N-2-fluorenylacetamide. Using hydroxylation of benzo[a]pyrene and N-2-fluorenylacetamide as a measure of mixed function oxidase activity, we found that the basal activities, which were 4- to 8-fold greater in the liver microsomes, were increased in both tissues after treatment of rats with the inducers. The induced activities were inhibited by 0.1 micrometers alpha-napthoflavone in vitro, indicating a dependence on cytochrome P 448. The data suggest that the mammary gland, an extrahepatic target for carcinogens, is capable of their metabolism. PMID- 6277342 TI - The influence of nicergoline on reaction kinetics of various forms of cyclic AMP phosphodiesterase. AB - 10-Methoxy-1,6-dimethylergoline-8 beta-methanol-5-bromonicotinate (nicergoline) has in vitro a modulatory effect upon reaction kinetics of various forms of cAMP phosphodiesterase (PDE) from beef heart and mice brain. Nicergoline inhibits low Km form of cAMP PDE from hear and brain; concentration of inhibitor required for 50% inhibition is 3-12 mumol/l. It is believe that the observed in vitro effects of nicergoline on reaction kinetics of various forms of cAMP PDE may in vivo help to normalize the level of cAMP, keep its concentration within certain limits, both by opposing large increase and large decrease in its cellular concentration. PMID- 6277343 TI - [The effect of the benzimidazole derivative AR-L 115 BS on the activities of myocardial adenylate cyclase and phosphodiesterase preparations (author's transl)]. AB - The effect of benzimidazole derivative 2-[(2-methoxy-4-methylsulfinyl)phenyl]-1H imidazol[4,5-b]pyridine (AR-L115 BS) on phosphodiesterase (PDE) and adenylate cyclase (AC) preparations from myocardial tissue was investigated in vitro. The following results were obtained: 1. AR-L 115 BS inhibits the PDE activity comparable to papaverine in a concentration-dependent and non-competitive way. Its inhibitory potency is about 40 times less if compared to papaverine since the Ki-values found are 315 and 7.9 mumol/l, respectively. The concentration of AR-L 115 BS producing a maximal positive inotropic effect on the isolated guinea-pig atrial preparation was shown to be 316 mumol/l, i.e., in the same concentration range as the biochemical findings. 2. The basal activity of myocardial AC preparations from reserpine pretreated guinea-pigs was not significantly altered in the presence of AR-L 115 BS concentrations ranging from 10 to 200 mumol/l. In contrast, the isoprenaline (1-100 mumol/l) stimulated AC was concentration dependently inhibited already by 1 to 10 mumol/l AR-L 115 BS. PMID- 6277344 TI - [Nerve biopsy. Advantages and limitations of modern examination techniques (author's transl)]. AB - The main technics of peripheral nerve biopsy examination, the diagnostic value of each of them and their limits are given. Chief pathological processes and changes and usual causes of peripheral neuropathies, the diagnosis of which can be provided by nerve biopsy are analysed. Most frequent errors and misleading changes are emphasized. An examination scheme is set up. PMID- 6277345 TI - Eosinophil mediated necrosis in a malignant fibrous histiocytoma. PMID- 6277346 TI - Simulation of anticipatory responses in classical conditioning by a neuron-like adaptive element. AB - A neuron-like adaptive element is described that produces an important feature of the anticipatory nature of classical conditioning. The response that occurs after training (conditioned response) usually begins earlier than the reinforcing stimulus (unconditioned stimulus). The conditioned response therefore usually anticipates the unconditioned stimulus. This aspect of classical conditioning has been largely neglected by hypotheses that neurons provide single unit analogs of conditioning. This paper briefly presents the model and extends earlier results by computer simulation of conditioned inhibition and chaining of associations. PMID- 6277347 TI - Lesions of the olfactory pathways affecting neophobia and learned aversion differentially. AB - The contribution of ascending olfactory pathways in neophobia and learned aversion to the same food was investigated in male rats bearing lesions of both olfactory peduncles, or one olfactory peduncle and the opposite lateral olfactory tract or anterior limb of the anterior commissure. The animals were fed on usual stock diet (S) offered as a choice with novel vanilla food (V) on test days: during neophobia (N), then before and after aversive conditioning (Aa, At). Daily food intake was measured, and the preference was expressed as V/(V + S). Experiment 1 included a neophobia test, before aversive conditioning (3 mEq/kg LiCl, i.p.). In Experiment 2, aversion only was studied (0.9 mEq/kg). In the neophobia test, the preference ratio was 7% in unoperated controls, and 43-52% in the 3 lesioned groups. The same controls had preference ratios equal to 64% and 22%, before and after aversive learning. Similar drops were observed for any lesioned group in Expt. 1. The decrease was less obvious, although significant, in rats of Expt. 2 with asymmetric lesions; those with both olfactory peduncles cut through maintained the same preference ratio (48%) before and after LiCl treatment. The data are interpreted assuming that: (1) lateral olfactory tract and anterior commissure both contribute to information processing in neophobia and aversion; (2) olfactory cues subserve neophobia prepotently; and (3) one cannot account for the sensory determinism of neophobia and aversion calling for a single mechanism. PMID- 6277348 TI - Altered behavioral responsivity to morphine during the periadolescent period in rats. AB - The results of previous studies have suggested that periadolescent rats are differentially affected by catecholaminergic agents, when compared with younger or adult animals. Since dopamine-opiate interactions have been demonstrated in rats, the present study was conducted to evaluate the behavioral responsivity of periadolescent and adult rats following doses of morphine ranging from 1 to 10 mg/kg i.p. The responses measured included matrix crossings, rearing, grooming, auto-directed stereotyped behavior and gnawing, and were recorded continuously from 30 to 90 min post-injection. When indicated, scores on each measure were transformed to control for differences observed in the baseline (saline) groups. Analyses of all measures revealed a significant effect of morphine dose, although the shape of the dose-response curve differed for individual responses. In addition, those measures which might be considered locomotor responses (matrix crossing, rearing), as well as grooming, revealed significant main or interactive effects of age. In contrast, those measures characterized as stereotyped behaviors (auto-directed stereotyped behavior and gnawing) revealed no differential effect in periadolescent animals relative to adults. Since locomotor responses and stereotyped behavior have been suggested by previous research to be mediated by the mesolimbic and extrapyramidal dopamine systems, respectively, these results further support the previously suggested hypothesis of the delayed ontogeny of mesolimbic relative to extrapyramidal dopamine pathways. PMID- 6277349 TI - Analysis of behavioural responses to an ACTH analog in CXB/By recombinant inbred mice. AB - Male mice of the C57BL/6By and BALB/cBy inbred strains, their reciprocal F1 hybrids, and 7 recombinant inbred strains, were tested for open-field activity, a shock-motivated successive reversal position discrimination problem in a T-maze, and a toggle box exploration task. The test battery was repeated one month later. Finally, mice were tested for the acquisition and extinction of a taste aversion conditioned by ethanol injection. Mice of each strain were tested after injection with saline or one of 3 doses of an ACTH analogue. Highly significant genotypic differences were found for all measures, an expected result. The strain distribution pattern seen in the toggle box suggested single gene mediation of exploratory activity after habituation. One aspect of avoidance responding and extinction of conditioned taste aversion also yielded strain distribution pattern consistent with single gene control. Peptide treatment reduced internal field crossings in the open field. This effect was not strain dependent. Peptide treatment had no effect on T-maze learning, conditioned taste aversion, or toggle box exploration. PMID- 6277350 TI - [Probenecid-induced convulsion and cerebrospinal cyclic nucleotides in the kindling cat preparations (author's transl)]. PMID- 6277352 TI - [The nucleolus-like bodies in the mouse locus coeruleus (author's transl)]. PMID- 6277351 TI - [Endocrinological analysis of pituitary adenomas associated with hyperprolactinemia (author's transl)]. PMID- 6277353 TI - Acetylcholine receptor. PMID- 6277354 TI - Clinical assessment of neuromuscular transmission. PMID- 6277355 TI - The seed and the soil: effect of dosage, personality and starting state on the response to delta 9 tetrahydrocannabinol in man. AB - 1 The effects of two doses of delta 9THC (2.5 and 10 mg), delivered by paced smoking of herbal cigarettes, on CNV magnitude, subjective mood ratings and heart rate were studied in 20 subjects. 2 There were highly significant interactions between drug dosage and Extraversion and Neuroticism scores, so that the direction and degree of response to the different doses of delta 9THC depended on the personality characteristics of the subjects. 3 The effects of 9 mg delta 9THC and placebo, delivered in herbal cigarettes smoked naturally, on smoking behaviour, subjective mood ratings, measures of autonomic activity and auditory and visual cortical evoked responses were compared in 12 subjects. 4 Smoking behaviour, subjective 'high' rating and elevation of heart rates were the most significant discriminators between drug and placebo. The latency of some of the components of the visual evoked responses was also increased by delta 9THC. 5 There was a significant correlation between the effects of delta 9THC on skin conductance reactivity and the basal (pre-drug) level, reactivity increasing after drug in subjects with low basal reactivity and decreasing in those with high basal levels. 6 Both experiments provided clear evidence of dose-dependent biphasic stimulant and depressant actions of delta 9THC on both subjective and objective measures, and these effects were influenced by the personality and the starting state of the subjects. PMID- 6277356 TI - Effects of the angiotensin converting enzyme inhibitor, captopril, in essential hypertension. AB - 1 The effects of the orally active angiotensin converting enzyme inhibitor, captopril, were examined in 15 patients with mild or moderate essential hypertension. 2 Following initial dosing with captopril 25 mg, there was a significant fall in supine and erect blood pressure in 2 h and lasting for 6 h. There was no significant alteration of heart rate. No reduction was seen in plasma noradrenaline concentration. Maximum inhibition of plasma converting enzyme activity occurred 60 min post-dosing. 3 There was a strong positive correlation between blood pressure reduction and converting enzyme inhibition. The magnitude of the blood pressure reduction and converting enzyme inhibition following initial dosing was reflected in subsequent success with captopril monotherapy during a 12 week follow-up period. 4 It was also found that patients who did not achieve adequate blood pressure control on captopril in doses of 50 mg three times daily or less still were not controlled on a dose of 150 mg three times daily without the addition of a diuretic. 5 The risks of renal and marrow toxicity from captopril may be reduced by administering only low doses with the addition, where necessary, of a thiazide diuretic. PMID- 6277357 TI - Ranitidine delays gastric emptying of solids in man. PMID- 6277358 TI - Effects of endocrine therapy on steroid-receptor content of breast cancer. AB - In order to determine the mechanisms of relapse following response to endocrine therapy, we have measured the oestrogen receptor (RE) content of biopsies of breast cancer in patients receiving various types of endocrine treatment. RE content fell in responding (means of 260.2 to 12 fmol/mg protein) and in nonresponding (means of 155.1 to 31.8 fmol/mg protein) patients who had measurable receptor at the start of treatment. Some of these patients, and a further group of responders to endocrine therapy, were monitored until relapse. Tumour biopsies at the time of relapse showed that 10/14 tumour samples contained significant RE (mean of 86.7 fmol/mg protein; range less than 10-271 fmol/mg protein) after successful endocrine therapy. No relationship could be found between RE content and plasma gonadotrophin or steroid-hormone concentration, but the fall in RE content correlated with reduced numbers of tumour cells in the biopsy. These results indicate that relapse following successful endocrine therapy in breast cancer does not appear to be due to the emergence of RE negative tumour cells. The fall in RE content during response to endocrine therapy may be due to reduced tumour-cell content of the biopsy. PMID- 6277359 TI - Circulating ACTH and related peptides in lung cancer. AB - The prevalence of high levels of circulating ACTH-like immunoactivity was determined in 134 patients with lung cancer, using reference ranges from 52 age- and sex-matched patients with non-malignant lung disease. Two studies used ACTH radioimmunoassays with different specificities. Study A used an unextracted plasma or serum assay for total ACTH immunoactivity. High serum ACTH levels occurred in 24% of patients with small-cell carcinoma and 3% of patients with non small-cell cancer. In patients with small-cell carcinoma, levels were high in 12% with limited disease and 32% with extensive disease. Study B used an ACTH assay after plasma extraction by porous glass, which measured mainly regular 1-39 ACTH. Here no lung-cancer patient had levels above the reference range, suggesting that the high levels in Study A may be due to plasma ACTH components which are poorly extracted by porous glass. It is concluded that high circulating ACTH immunoactivity occurs in a minority of patients with lung cancer, particularly those with extensive small-cell carcinoma. Indirect evidence suggests that the high ACTH levels detected with assays for total ACTH are due to molecular forms other than 1-39 ACTH, probably high-mol.-wt species. PMID- 6277360 TI - The role of penicillin in the pathogenesis of chronic urticaria. AB - Intracutaneous tests with penicilloyl-polylysine (PPL) and benzyl-penicillin G (PG) were performed in 245 patients suffering form chronic recurrent urticaria, including physical urticaria. Positive results were observed in fifty-nine patients (24%). Sera from fifty-seven of these fifty-nine patients were investigated for circulating anti-penicilloyl antibodies by an enzyme-linked immunosorbent assay (ELISA) and a passive haemagglutination test (HA). The results were compared to those form the ELISA and HA in a control group of thirty five patients who had shown clinical allergic reactions to penicillin and had positive skin tests to PPL and/or PG. The in vitro tests revealed positive results in 12.3% and 37.1% respectively. In forty-three patients the course of the positive intracutaneous tests to penicillin, together with the duration of chronic urticaria, was followed over a period of time up to 3.5 years. In twenty two out of forty-two patients with positive intracutaneous tests to penicillin, a diet free of dairy products proved to have a curative effect, compared to two out of forty control subjects with chronic urticaria and negative skin tests to penicillin. These studies indicate that penicillin has an important role in the aetiology and maintenance of chronic urticaria. PMID- 6277361 TI - DNA chain elongation rates in marrow cells from vitamin B12-deficient patients and methotrexate-treated mice. AB - The DNA synthesized by marrow cells from patients with vitamin B12 deficiency and mice previously given methotrexate (MTX), has been investigated. Suspensions of bone marrow cells were pulse-labelled with [methyl-3H]thymidine or deoxy [5 3H]cytidine for 30 s and the radioactivity in the DNA was chased thereafter in the presence of 10 microM non-radioactive nucleoside for periods up to 60 min. The rates of elongation of new daughter strands were then assessed by hydroxyapatite chromatography of alkali-denatured DNA samples. No significant differences were found between the average rates of elongation of daughter strands from control marrow cells on the one hand and the vitamin B12-deficient or the methotrexate-affected cells on the other. This is to be contrasted with the results of previous studies which have shown a retardation in the rates of movement of replication forks in stimulated, cultured lymphocytes obtained from vitamin B12- or folate-deficient patients. PMID- 6277362 TI - The role of the enterohepatic cycle in folate supply to tumour in rats. AB - The importance of the folate enterohepatic cycle in governing the supply of folate to implants of a rapidly-growing tumour were studied in a new animal model. Following enteric administration of tritiated pteroylglutamic acid, [3H]PteGlu1, tumour uptake of labelled folate was limited to CH3[3H]H4PteGlu1 produced by the gut mucosal cells during absorption or subsequently recirculated through the enterohepatic cycle. 50% of the labelled folate reaching the tumour nodules in the first 6 h after enteric administration first circulated through the enterohepatic cycle. In addition, labelled folate taken up by tumour was immediately incorporated into a polyglutamyl folate pool. There was no evidence for a release of labelled folate from tumour for recirculation to the liver. Therefore the liver and folate enterohepatic cycle appear to play a major role in regulating the supply of folate to rapidly proliferating tissues such as tumour by acutely storing folate from the diet and then secreting it into bile for reabsorption and transport to tissue. PMID- 6277363 TI - Circulating placental proteins (hCG, SP1 and PP5) in trophoblastic disease. AB - Serum human chorionic gonadotrophin (hCG), pregnancy-specific beta 1-glycoprotein (SP 1) and placental proteins 5 (PP5) levels have been measured by radioimmunoassay in 20 patients (116 samples) with hydatidiform mole, one patient (nine samples) with invasive mole and 10 patients (103 samples) with choriocarcinoma. Measurement of both serum SP1 and hCG are useful in the monitoring of these diseases. The presence of PP5 in hydatidiform mole and its absence in choriocarcinoma support the hypothesis that PP5 is closely associated with the invasive activity of malignant trophoblast. PMID- 6277364 TI - Stereospecificity of (E)- and (Z)-phosphoenol-alpha-ketobutyrate with chicken liver phosphoenolpyruvate carboxykinase and related phosphoenolpyruvate-utilizing enzymes. PMID- 6277365 TI - Thyroid hormone binding to human serum prealbumin and rat liver nuclear receptor: kinetics, contribution of the hormone phenolic hydroxyl group, and accommodation of hormone side-chain bulk. PMID- 6277366 TI - Specificity and binding affinity of phospholipids to the high-affinity cardiolipin sites of beef heart cytochrome c oxidase. PMID- 6277367 TI - Adenylate kinase of Escherichia coli: evidence for a functional interaction in phospholipid synthesis. AB - Previous genetic and biochemical experiments have suggested that the adenylate kinase of Escherichia coli may be directly involved in phospholipid synthesis through formation of a complex with sn-glycerol-3-phosphate acyltransferase, the membrane-bound enzyme that catalyzes the first step in phospholipid synthesis. In this paper we report direct experiments to test this hypothesis. A mutation within the adenylate kinase structural gene is described that results in a temperature-sensitive phospholipid synthesis (assayed in vivo) and a temperature sensitive acyltransferase. The adenylate kinase activity of this strain is only minimally altered either in vitro or [as assayed by adenosine 5'-triphosphate (ATP) levels] in vivo. This result demonstrates that the inhibition of phospholipid synthesis is not the result of reduced ATP levels. We report the purification of E. coli adenylate kinase to homogeneity; and find that the addition of homogeneous wild-type adenylate kinase to membranes containing a mutationally altered temperature-sensitive acyltransferase results in thermal stabilization of the acyltransferase activity. Ovalbumin has no such protective effect. Purified E. coli inner membranes contain several proteins that are precipitated by addition of anti adenylate kinase antibody to detergent solubilized membranes. PMID- 6277368 TI - Differential effect of hypertonic initiation block on the synthesis of collagen chains by cultured chick embryo cells. AB - The major type of collagen synthesized by fibroblasts or bone cells, type I collagen, consists of two chains normally found in a 2:1 ratio designated alpha 1(I)2 alpha 2(I) or more simply alpha 1(I)2 alpha 2. I have analyzed the relative synthesis of type I chains in these cells under conditions which reduce the initiation of protein synthesis. It was found that in bone cells, which make a large amount of collagen, the alpha 1(I):alpha 2 ratio is unaltered whereas in fibroblasts, which make smaller amounts of collagen, the alpha 2 chain is particularly sensitive to these same conditions. Examination of the collagen secreted into the medium, under these same conditions, also revealed an altered chain ratio from cells making low amounts of collagen. PMID- 6277369 TI - Phosphoenolpyruvate-dependent fructose phosphotransferase system of Rhodopseudomonas sphaeroides: purification and physicochemical and immunochemical characterization of a membrane-associated enzyme I. AB - The phosphotransferase system (PTS) of the phototrophic bacterium Rhodopseudomonas sphaeroides consists of a component located in the cytoplasmic membrane and a membrane-associated enzyme called "soluble factor" (SF) [Saier, M. H., Feucht, B. U., & Roseman, S. (1971) J. Biol. Chem. 246, 7819--7821]. SF has been partially purified by a combination of hydrophobic interaction and ion exchange and gel-permeation chromatography. SF is similar to Escherichia coli enzyme I in its molecular characteristics and enzymatic properties. It has a molecular weight of 85 000 and readily dimerizes. Phosphoenolpyruvate and Mg2+ stabilize the dimer. The enzyme catalyzes the conversion of phosphoenolpyruvate into pyruvate and becomes phosphorylated in the process. The phosphoryl group is subsequently transferred to fructose in the presence of R. sphaeroides membranes. SF substitutes for E. coli enzyme I in fructose or glucose phosphorylation with E. coli enzyme II and HPr. The activities of SF with the R. sphaeroides PTS and the E. coli PTS reside on structurally distinct molecules as shown by their response to limited proteolytic digestion and by immunochemical studies. The activity of SF with the E. coli PTS arises during the isolation procedure and is most likely due to the removal of HPr-like protein from SF. PMID- 6277370 TI - Calcium channels in the neuronal membrane. PMID- 6277371 TI - pH homeostasis in bacteria. PMID- 6277372 TI - Ion transport in yeast. PMID- 6277373 TI - The location of redox centers in biological membranes determined by resonance x ray diffraction. I. Observation of the resonance effect. AB - We have developed resonance X-ray diffraction methods to locate for the first time intrinsic metal atoms associated with redox centers within biological membrane systems. The study of membranes containing dilute concentrations of resonant scatterers has been made possible by the development of synchrotron radiation sources of X-rays. The technique permits altering the scattering power of a particular atom relative to others by varying the incident X-ray energy. Thus, this method may be used to locate a metal atom within a complex integral protein without chemical modification of the membrane. We present resonance diffraction data taken with synchroton radiation for two different membrane systems: cytochrome oxidase incorporated into lipid vesicles and a photosynthetic reaction center-cytochrome c complex also reincorporated into lipid vesicles. PMID- 6277374 TI - The location of redox centers in biological membranes determined by resonance x ray diffraction. II. Analysis of the resonance diffraction data. AB - In the preceding paper (Stamatoff, J., Eisenberger, P., Blasie, J.K., Pachence, J.M., Tavormina, A., Erecinska, M., Dutton P.L. and Brown, G. (1982) Biochim. Biophys. Acta 679, 177-187), we described the observation of resonance X-ray scattering effects from intrinsic metal atoms associated with redox centers in membrane proteins on the lamellar X-ray diffraction from oriented multilayers of reconstituted membranes. In this paper, we discuss the possible methods of analysis of such data and present the results of our model refinement analysis concerning (a) the location of the cytochrome c heme iron atom in the profile structure of a reconstituted membrane containing a photosynthetic reaction center cytochrome c complex and (b) the location of the heme a and a3 iron atoms in the profile structure of a reconstituted membrane containing cytochrome oxidase. The former results are of special importance because they provide a test of the validity of the resonance diffraction data and the methods of analysis, since the location of cytochrome c in the reaction center-cytochrome c membrane profile is known independently of the resonance diffraction experiments. PMID- 6277375 TI - Vesicular preparation of a highly coupled ATPase-ATP synthase complex from pig heart mitochondria. AB - 1. A method is described to prepare an ATPase-ATP synthase complex from pig heart mitochondria exhibiting a very high ATP-32Pi exchange activity (1.6 mumol/min per mag protein in optimal conditions). 2. The preparation is virtually devoid of nucleoside diphosphokinase and adenylate kinase activities. 3. Freeze-fracture studies show that the ATPase-ATP synthase complex is integrated in lipid vesicles of 400-600 A in diameter. 4. It contains the endogenous natural proteic inhibitor which seems to behave as a coupling factor. 5. The rate of ATP hydrolysis catalyzed by the ATPase-ATP synthase complex is competitively inhibited by ADP, while the presence of ADP increases the initial rate of 32Pi incorporation into ATP. 6. The 32Pi incorporation into ATP can occur at a rate almost equal to that of nucleoside triphosphate (NTP) hydrolysis provided that the rate of NTP hydrolysis is kept low and that the ADP concentration is high enough. In these conditions, a very high coupling between NTP hydrolysis and ATP synthesis can be demonstrated. PMID- 6277376 TI - Diglyceride kinase from Escherichia coli. Modulation of enzyme activity by glycosphingolipids. AB - Diglyceride kinase was purified from membranes of Escherichia coli K-12 using organic solvents. The enzyme apoprotein depended on lipids, such as cardiolipin (diphosphatidylglycerol), phosphatidylcholine or 1-monooleoylglycerol, for activity with 1,2-dipalmitoylglycerol. Mixed brain cerebrosides and gangliosides as well as defined ganglioside fractions and synthetic lactocerebroside were devoid of lipid cofactor activity. However, all these glycosphingolipids were strong inhibitors of activation by phosphatidylcholine. When cardiolipin was used as lipid activator with the detergent, Triton X-100, as solubilizing agent, the addition of mixed or purified gangliosides first (at about 0.4 mM) resulted in additional activation, but higher ganglioside concentrations were strongly inhibitory. Both effects were absolutely dependent on the presence of lipid-bound sialic acid and were not given by cerebrosides, by free sialic acid or by sialyl lactose. The stimulating and inhibitory effects of glycosphingolipids could also be demonstrated when 1-monooleoylglycerol was used as substrate, lipid activator and solubilizing agent at the same time. The modulation of kinase activity by glycosphingolipids is discussed at the level of lipid/protein interactions. PMID- 6277377 TI - Alkaline phosphodiesterase I and alkaline phosphatase I in plasma membranes of herpes simplex virus type 1 transformed hamster cells. AB - Plasma membrane extracts from Herpes simplex virus type 1 transformed hamster embryo fibroblasts were chromatographed on Lens culinaris lectin coupled to Sepharose (LcH-Sepharose) and analysed by dodecyl sulphate polyacrylamide gel electrophoresis. Coomassie blue-staining revealed two major protein bands with apparent molecular weights of 125 000 and of about 75 000-90 000. In plasma membranes isolated from these tumor cells prior labeled with [3H]fucose or [3H]glucosamine these bands contained the highest amounts of incorporated radioactivity. Separation by LcH-Sepharose-affinity chromatography as well as metabolic labeling clearly demonstrates their glycoprotein character. The 125 000 protein coincides with alkaline phosphodiesterase I activity with a Km of 6 . 10( 4) M for TMP p-nitrophenyl ester and is competitively inhibited by UDP-N acetylglucosamine. This enzymatic activity is also present in normal hamster embryo fibroblasts. Gel electrophoresis of the Lens culinaris lectin-binding glycoproteins from plasma membranes of normal hamster embryo fibroblasts additionally revealed a strong alkaline phosphatase activity represented by an apparent molecular weight of 150 000, while HSV1 hamster tumor cells contain only a very weak activity of this enzyme activity. HSV-lytically infected cells, however, have unchanged levels of alkaline phosphatase activity, whereas alkaline phosphodiesterase activity increases slightly. PMID- 6277378 TI - Characteristics of high-affinity [3H]adenosine binding to rat brain synaptosomes and turkey erythrocyte membranes. AB - High affinity binding sites for [3H]adenosine in rat brain and in turkey erythrocytes can be identified by binding experiments. Displacement experiments using a number of adenosine analogs indicate that these high affinity sites do not represent the R-type adenosine receptors which mediate activation of adenylate cyclase, although the binding is theophylline sensitive. Similarly, the binding of [3H]adenosine is not to the P-site, which mediates inhibition of adenylate cyclase, since the high affinity binding persists in the presence of 2',5'-dideoxyadenosine. Furthermore, these results remain qualitatively similar also in the presence of dipyridamole which blocks adenosine transport sites. We conclude that theophylline sensitivity does not indicate that [3H]adenosine binding sites correspond to adenosine receptors coupled to adenylate cyclase. PMID- 6277379 TI - Adriamycin inhibits the formation of non-bilayer lipid structures in cardiolipin containing model membranes. AB - The effect of adriamycin on cardiolipin-containing model membrane systems have been studied by 31P-NMR, freeze-fracture electron microscopy and binding experiments. Adriamycin effectively inhibits the formation of non-bilayer lipid structures induced by Ca2+ and cytochrome c in cardiolipin-containing liposomes. This drug also strongly inhibits the uptake of Ca2+ by cardiolipin into an organic phase. These results are discussed in relation to the cardiotoxic effect of adriamycin and the possible importance of non-bilayer lipid structures for the functioning of the mitochondrion. PMID- 6277380 TI - Reversal of copper-induced inhibition of vasopressin responsiveness by reducing agents. AB - Copper inhibits the hydro-osmotic response to vasopressin in the urinary bladder of Bufo marinus at a site proximal to cyclic AMP production. This effect is not reversed by washing in Cu2+ -free Ringer's solution but is overcome by serosal addition of reducing agents, suggesting that vasopressin responsiveness in this tissue is modulated by either the redox potential or the sulfhydryl content of the serosal membrane. PMID- 6277381 TI - Pyrophosphate analogues as inhibitors of DNA polymerases of cytomegalovirus, herpes simplex virus and cellular origin. AB - Several pyrophosphate analogues have been compared for their ability to inhibit the activities of isolated cytomegalovirus (CMV) DNA polymerase, herpes simplex virus type 1 (HSV 1) DNA polymerase and calf thymus DNA polymerase alpha. The most effective inhibitors were phosphonoformate and phosphonoacetate. Although not identical, the structural requirements for compounds inhibitory to CMV and HSV-1 DNA polymerase were specific, with two negatively charged groups in close vicinity. The CMV DNA polymerase was more susceptible to certain phosphonoacetates containing bulky hydrophobic alpha-substituents than was the HSV-1 DNA polymerase. No example of the converse preference was found. The inhibition of CMV DNA polymerase by phosphonoformate, hypophosphate, alpha hydroxyphosphonoacetate and alpha-nonylphosphonoacetate was linear non competitive with the deoxyribonucleoside triphosphates as variable substrates. Phosphonoformate, phosphonoacetate, and to a lesser extent alpha hydroxyphosphonoacetate, carbonyldiphosphonate and alpha-nonylphosphonoacetate also inhibited the focus formation by CMV in cell-culture. PMID- 6277382 TI - Purification and properties of two endonucleases specific for apurinic/apyrimidinic sites in DNA from Saccharomyces cerevisiae. AB - Two distinct endonucleases from Saccharomyces cerevisiae, specific for apurinic/apyrimidinic sites (AP-endonucleases A and B), have been extensively purified and characterized. Both are free from unspecific and ultraviolet specific endonucleases and exonucleases. The two enzymes are monomeric proteins of around 24000 daltons. Both are sensitive to ionic strength and most active in the presence of 150 and 100 mM NaCl for AP-endonucleases A and B, respectively. They are not absolutely dependent on divalent cations, since they are insensitive to EDTA, although AP-endonuclease A is activated by Ca2+ or Mg2+ and AP endonuclease B by Mg2+ only. ATP inhibits the enzymes. AP-endonuclease A reacts optimally between pH 6 and 8, and AP-endonucleases B at pH 8. AP-endonuclease A is more stable at 60 degree C (half-life of 17 min) than B (half-life of 4 min). AP-endonuclease A is insensitive to N-ethylmaleimide or rho chloromercuribenzoate. AP-endonuclease B is also insensitive to N-ethylmaleimide, but rho-chloromercuribenzoate inhibits its activity. PMID- 6277383 TI - Regulation of ornithine decarboxylase activity by cyclic AMP in guinea pig lymphocytes: transcriptional and post-transcriptional control. AB - Induction of ornithine decarboxylase (L-ornithine carboxy-lyase, EC 4.1.1.17) in phytohemagglutinin-stimulated lymphocytes was suppressed by the administration of dibutyryl cyclic AMP and 3-isobutyl-1-methylxanthine. This inhibition of ornithine decarboxylase induction was overcome by removing the chemicals from the lymphocyte culture. However, the recovery of ornithine decarboxylase induction by washout of the chemicals did not occur when actinomycin D, cordycepin or alpha amanitin was added to the cultures, suggesting that the synthesis of mRNA directed to ornithine decarboxylase was reduced in dibutyryl cyclic AMP- and 3 isobutyl-1-methylxanthine-treated cells. Moreover, the decay of ornithine decarboxylase activity in the cells treated with phytohemagglutinin plus dibutyryl cyclic AMP and 3-isobutyl-1-methylxanthine was significantly slower than that of control cells stimulated with phytohemagglutinin alone. The difference of the decrease in ornithine decarboxylase activity between dibutyryl cyclic AMP- and 3-isobutyl-1-methylxanthine-treated and control cells was observed when actinomycin D was added to the culture, but not when cycloheximide was added. Therefore, prevention of ornithine decarboxylase decay by cyclic AMP may be post-transcriptional events, but not post-translational events. PMID- 6277384 TI - Studies on the function of the non-primer tRNAs associated with the 70 S RNA of avian myeloblastosis virus. AB - Significant amounts of three tRNAs are associated with the 70 S RNA of avian myeloblastosis virus (AMV). The temperatures at which they are half dissociated from the 70 S RNA in 50 mM NaCl and their respective quantities relative to 35 S RNA are: tRNAArg, 51 degree C, 1.6; tRNALys, 57 degree C, 0.7 and tRNATrp, 76 degree C, 1.0. Possible functions for the non-primer tRNAs (tRNAArg and tRNALys) were evaluated by determining the effect of their thermal dissociation on: (a) conversion of 70 S to 35 S RNA, (b) capacity of 70 S and/or 35 S RNA to be translated in vitro, and (c) capacity of 70 S and/or 35 S RNA to be reverse transcribed in vitro. Conversion of 70 S to 35 S RNA occurred with a tm of 56 degree C and is consistent with the hypothesis that tRNALys might be involved in joining two 35 S RNA subunits to form the 70 S RNA complex. There was no indication that the association of either tRNAArg or tRNALys influenced the rate or quality of translation of 70 S or 35 S RNA. A decrease in the rate at which 70 S RNA is transcribed occurs in parallel with the dissociation of tRNAArg and tRNALys. PMID- 6277385 TI - Rat ovarian nuclear protein kinases. PMID- 6277386 TI - Regulation of protein kinases activity by quercetin in Ehrlich ascites tumor cells. AB - Cyclic AMP-independent protein kinase activities from Ehrlich ascites tumor cells, partially purified by DEAE-cellulose and phosphocellulose chromatography were inhibited by quercetin. The cyclic AMP in the tumor ascites cells and the cyclic AMP-dependent protein activity from this tumor and from bovine and mouse tissues were unaffected by this drug. Since we reported that quercetin elevates cyclic AMP level in Ehrlich ascites tumor cells, this bioflavonoid may have a dual effect on the protein kinase activities in these cells, thus, increasing the cyclic AMP-dependent and decreasing the cyclic AMP-independent protein kinase activities. PMID- 6277387 TI - A critical interpretation of experiments on binding of peptide hormone to specific receptors by computer modelling. AB - Many investigations dealing with the interaction of peptide hormones and specific cell membrane receptors imply the existence of two classes of independent binding sites. On class is characterized by high affinity and low capacity, the other one by low affinity and high capacity. This conclusion has been derived from the fact that the Scatchard plots of binding data show a significant upward curvature. Using a more precise and critical method of evaluation these findings probably must be revised in some cases. Other conclusions taken from linear plots concerning the regulation of hormone receptors should be discussed more carefully. Some sources of errors caused by an uncritical interpretation of Scatchard plots are demonstrated. PMID- 6277388 TI - A quantitative analysis of glucagon binding to isolated intact neonatal and adult rat hepatocytes on the basis of two different binding models. AB - The interaction of glucagon with specific receptors has been studied in isolated intact neonatal and adult rat hepatocytes. The hormone binding measured directly with 125I-labelled glucagon was saturable and reversible. The 125I-labelled glucagon binding was inhibited by unlabelled homologous hormone at concentrations ranging from 0.5 nM to 50 microM. Two different binding models were assumed to analyse the binding data by a nonlinear least-squares procedure: (I) a single class of independent sites and (II) two classes of independent sites. The comparison of the fitted theoretical curves reveals that both binding models are in fact compatible with these data. Adult hepatocytes have a considerably higher affinity for glucagon than neonatal hepatocytes; the binding capacity of neonatal liver cells from 1-7-days-old rats proved to be markedly reduced compared with the cells from adult rats. The glucagon-induced intracellular cyclic AMP production was measured at various hormone concentrations under conditions identical to those for the determination of extracellular hormone binding. The correlation of both parameters indicates a direct connection between receptor occupancy and adenylate cyclase stimulation of both parameters indicates a direct connection between receptor-occupancy and adenylate cyclase stimulation. These results suggest that a decreased receptor concentration in neonatal hepatocytes is responsible for the decreased cyclic AMP production. PMID- 6277389 TI - High-affinity binding of 2,3,7,8-tetrachlorodibenzo-p-dioxin in cell nuclei from rat liver. AB - The intranuclear binding of radioactive 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) in rat liver has been studied both in vivo and in vitro. Following the intravenous administration of [1,6-3H]TCDD, a maximum uptake by cell nuclei could be observed at 2 h after injection with a concurrent decrease in the cytosolic uptake. Using linear sucrose density gradient centrifugation, dextran-coated charcoal adsorption assay, DEAE-Sepharose ion-exchange chromatography, competition, enzymatic and saturation studies, a high-affinity binding protein for TCDD in liver cell nuclei could be demonstrated both in vivo and after an exchange in vitro of intravenously administered unlabelled 2,3,7,8- tetrachlorodibenzofuran (TCDBF) for [3H]TCDD. Sucrose density gradient analysis showed a size of 4-5 S for both the cytosolic and nuclear TCDD binding entity. The specific binding of [3H]TCDD to nuclear components was heat labile and saturable and had an equilibrium dissociation constant of 1.05 nM. Based on a differential susceptibility to specific hydrolases, i.e. DNAase, RNAase, trypsin and pronase, the binding entity appears to be a 4-5 S salt-extractable protein. PMID- 6277390 TI - 5'-nucleotidase activity in permanent human lymphoid cell lines. Implication for cell proliferation and aging in vitro. PMID- 6277391 TI - Binding of fibrinogen molecules to pig platelets and their membranes. AB - Following addition of ADP, 125I-labelled fibrinogen binds specifically to pig platelets. This binding is completely inhibited by the unlabelled fibrinogen. Quantitative analysis indicates the presence of 12,400-25,000 molecules of fibrinogen which can be bound with an association constant of 5 . 10(8) M-1 to platelets. Fibrinogen receptors were found to be active in the isolated platelet membranes as well. Quantitative analysis of the saturable binding of fibrinogen to the platelet membranes showed that these receptors react with the same affinity with fibrinogen molecules. In contrast to the intact platelets, the platelet membranes can specifically bind fibrinogen in the absence of ADP. We conclude that a specific receptor for fibrinogen is exposed on the surface as a result of cell damage which is the first step of the platelet membrane isolation. PMID- 6277392 TI - Inhibitors of membrane depolarization regulate acetylcholine receptor synthesis by a calcium-dependent, cyclic nucleotide-independent mechanism. AB - The inhibition of membrane depolarization by tetrodotoxin or the local anesthetic benzocaine elevates the acetylcholine receptor levels in cultured myotubes. The elevated acetylcholine receptor levels are due to increased receptor synthesis rather than to decreased degradation. The effects of tetrodotoxin and benzocaine on acetylcholine receptor levels are not additive, and are not inhibited by exogenously added cyclic GMP analogues or by elevated intracellular levels of cyclic GMP. However, the stimulation of acetylcholine receptor levels by tetrodotoxin or benzocaine is reversed by the addition of the calcium ionophore A23187. In contrast, tetrodotoxin or benzocaine stimulated acetylcholine receptor synthesis beyond the maximal stimulation produced by cholera toxin. These results suggest that the inhibition of membrane depolarization elevates acetylcholine receptor synthesis by a calcium-dependent, cyclic nucleotide-independent mechanism. PMID- 6277393 TI - Thyrotropin controls cyclic nucleotide metabolism of thyroid follicular cells without affecting membrane potential or input resistance. AB - The action of thyrotropin on the rat thyroid follicular cell has been investigated using continuous transmembrane potential and input resistance recording from individual cells in in vitro preparations. The membrane potential in this study was -68.0 mV +/- 0.6 (mean+/-S.E.). Over a wide range of concentrations (1.5-50 mU/ml), thyrotropin failed to affect membrane potential or input resistance while 20 mU/ml thyrotropin was shown to elicit complex time dependent changes in tissue levels of both cyclic AMP and cyclic GMP. The present results reveal that thyrotropin-receptor interaction does not affect plasma membrane permeability, but is characterized by complex changes in endogenous cyclic nucleotide metabolism. PMID- 6277394 TI - Spin label studies on osmotically-induced changes in the aqueous cytoplasm of Phaeodactylum tricornutum. AB - The effects of hyperosmotic stress and adaption on the aqueous cytoplasm of Phaeodactylum tricornutum have been studied with spin labels using 0.2M external Ni2+ to obtain spectra solely from labels within the cells. From partitioning of the TEMPO spin label between the internal aqueous phase and the membrane it is found that the internal volume of the cells decreased by approx. 50-60% in media of high osmotic strength (1.9 osmol/l). During the accumulation of proline in the cells (8.8 mg/ml packed cells) on incubation in the medium of high osmolarity for 3 days, the recovery of the volume was 80%. Further addition of proline to the medium resulted in an increase in the proline concentration in the cells (12.2 mg/ml packed cells) and a recovery in volume of 90%. Cells incubated in the absence of any nitrogen source showed very little recovery and were in a stressed state even in the absence of an osmotic gradient. From the rotational correlation times of the TEMPONE spin label it was found that the effective microviscosity in the cytoplasm of normal cells (approx. 3-8 cP) was considerably higher than that of the external medium (1 cP) and increased 1.5-2-fold under high osmotic stress (1.9 osmol/l). Adaption during the accumulation of proline only decreased the effective microviscosity by approx. 50% of the stressed-induced increase, a considerably smaller recovery than that of the cell volume. PMID- 6277395 TI - Determination of dinucleoside 5', 5"'-P1, P4- tetraphosphates by 31P and 1H NMR spectroscopy. PMID- 6277396 TI - Lipid-lipid and lipid-protein collision rates in membranes. Determination by evaluation of spin-spin interactions between 15N and 14N spin labels. PMID- 6277397 TI - EPR studies on the thermal stability of spin labeled ankyrin and protein 4.1 prepared from human erythrocyte membranes. PMID- 6277398 TI - Conformational analysis of corticotropin (ACTH) and conformation-activity relationship. PMID- 6277399 TI - Mossbauer spectroscopy and perturbed angular correlation studies of the rare-and alkaline-earth bone uptake. AB - Emission Mossbauer spectra and Perturbed Angular Correlation measurements have been performed on samples of mineral bone powder labelled with 161Tb3+ or 133Ba2+ ions after either in vitro absorption or uptake by metabolic pathway. The study of these hyperfine spectra, compared with those carried out when 161Tb or 133Ba are situated in either hydroxyde lattice or phosphate one, shows that the uptake modes of rare-and alkaline-earth ions on the bone matrix are different. The rare earth ion seems to be absorbed on the surface bone in an environment of hydroxyl groups similar to the structure of a rare earth hydroxyde. The alkaline earth ion bone uptake appears more complicated and would make according to the following process : at first, surface absorption on the hydroxylapatite in a hydroxyde environment and then cationic exchange with the calcium phosphate groups into bone crystals. PMID- 6277400 TI - Structural approach of the electron transfer pathways in multiheme cytochromes c3. PMID- 6277401 TI - Anomalous potassium channel-gating rates as functions of calcium and potassium ion concentrations. AB - With near normal monovalent ionic concentrations, the rate of increase of the potassium conductance after a depolarizing voltage-clamp step is slowed when the external calcium concentration is increased. This trend in the rise-time with changes in calcium is reversed when the axointernal potassium concentration is reduced (150 mM) and the periaxonal concentration is increased (50 mM); that is, the rise-time decrease with increasing calcium concentration. Furthermore, the degree of sigmoidality of the K-conductance time-course always increase when the rise-times increase for a given test potential. In the case of calcium surface charge screening, these effects may be caused by a shifted distribution of K-ions within the channels following the altered ion gradient, and by a consequent shift in the reciprocal electrostatic interactions between the ionic charges and channel-gate charges. PMID- 6277403 TI - Photoelectric currents across planar bilayer membranes containing bacterial reaction centers. Response under conditions of single electron turnover. AB - Light-induced electric current and potential responses have been measured across planar phospholipid membranes containing reaction centers from the photosynthetic bacterium Rhodopseudomonas sphaeroides. Under conditions in which the reaction centers are restricted to a single electron turnover, the responses can be correlated with the light-induced electron transfer reactions associated with the reaction center. The results indicate that electron transfer from the bacteriochlorophyll dimer to the primary ubiquinone molecule, and from ferrocytochrome c to the oxidized dimer occur in series across the planar membrane. Electron transfer from the primary to secondary ubiquinone molecule is not electrogenic. PMID- 6277402 TI - Solvent substitution as a probe of channel gating in Myxicola. Effects of D2O on kinetic properties of drugs that occlude channels. AB - The effects of solvent substitution on the steady-state and kinetic properties of drugs (gallamine triethiodide) and ions (nonyltriethylammonium and Ba++) known to occlude Na+ and K+ channels have been examined and compared with the effects of D2O on unmodified channels. In general, we observed large isotope effects on the kinetics of occlusion at temperatures of 5 degrees C, but only minor effects at 15 degrees C, consistent with processes involving significant solvent interaction. Steady-state behavior was not affected. In the case of gallamine, where a dual effect on INa is evident, although both processes were D2O sensitive, only the occlusion phase had a significant temperature dependence. PMID- 6277405 TI - Biophysical Society. 26th annual meeting. February 14-17, 1982, Boston, Massachusetts. Abstracts. PMID- 6277404 TI - A comparison of resolution-enhancement methods in saturation-transfer EPR. 15N isotopically substituted spin labels and 35 GHz high-frequency operation. AB - The use of (15)N isotopic substitution and 35 GHz (Q-band) operation as resolution enhancement methods in saturation-transfer electron paramagnetic resonance (ST-EPR) are compared. We find that both methods offer roughly comparable enhancements in ST-EPR resolution. The most powerful approach to resolving complex ST-EPR spectral behavior, however, will probably be the combined use of multiple-frequency X- and Q-band operation with (15)N isotopically substituted spin labels. PMID- 6277406 TI - Change in conformation of various DNAs on melting as followed by circular dichroism. PMID- 6277407 TI - Single dose kinetics of deuterium labelled delta 1-tetrahydrocannabinol in heavy and light cannabis users. AB - Deuterium labelled delta 1-tetrahydrocannabinol was administered intravenously (5.0 mg) and by smoking (10.0 mg) to five heavy and four light marihuana users. All subjects smoked an estimated amount of 8.6-9.9 mg delta 1 tetrahydrocannabinol. The plasma levels of delta 1-tetrahydrocannabinol were followed for 48 hours and in two subjects fof 72 hours after administration. The systemic availability after inhalation calculated from the area under curve values was in the range of 27 +/- 10% for the heavy users and 14 +/- 1% for the light users. There was little difference between the groups with regard to the amount of smoked delta 1-tetrahydrocannabinol or plasma levels and area under curve values obtained after i.v. administration. Thus, it seems likely that the statistically significant difference in systemic availability of smoked delta 1 tetrahydrocannabinol was due to a more efficient smoking by the heavy users. It is also indicated that heavy users prefer slightly higher delta 1 tetrahydrocannabinol plasma levels than light users. Based on the area under curve values after i.v. administration, a plasma clearance of 760-1190 ml min-1 was calculated. The elimination half-life of delta 1-tetrahydrocannabinol is more than 20 hours. The present results do not suggest that tolerance or sensitivity to delta 1-tetrahydrocannabinol in heavy users is readily achieved. PMID- 6277408 TI - The role of metabolism in the control of cyclic AMP efflux, cyclic AMP content, and insulin secretion from rat islets of Langerhans. AB - Glucose, alpha-ketoisocaproate, and N-acetylglucosamine all initiate insulin release and stimulate cyclic AMP efflux and cyclic AMP accumulation. Mannoheptulose inhibited these islet responses to glucose but not to N acetylglucosamine or alpha-ketoisocaproate. On the other hand, menadione inhibited these responses to all three secretagogues. Nicotinamide enhanced the insulin release due to glucose and alpha-ketoisocaproate and also enhanced cyclic AMP efflux and intracellular cyclic AMP accumulation. N-acetylglucosamine mediated insulin release, cyclic AMP efflux, and cyclic AMP accumulation were unaffected by nicotinamide. These results suggest that the metabolic state of the islet is a primary determinant of the ability of a secretagogue to raise cyclic AMP efflux and intracellular cyclic AMP, and these actions may be mediated in part by alterations in the redox state of the pyridine nucleotide system. PMID- 6277409 TI - Carbohydrate transport in Clostridium pasteurianum. AB - Clostridium pasteurianum is shown to possess constitutive PEP-dependent phosphotransferase systems transporting glucose, fructose, sucrose, sorbitol, and mannitol. With the exception of those for mannitol and sorbitol, each system is specific for a single substrate. At the end of growth, sporulation occurred and there was a coordinate increase in the activity of all the phosphotransferase systems. This increase occurred independently of the nature of the growth substrate. PMID- 6277410 TI - Identification of primary lysosomes in human megakaryocytes and platelets. AB - The presence of lysosomal enzymes in human platelets is well documented; the identity of the "lysosome," however, has been the subject of some disagreement. In order to determine the time of appearance and subcellular localization of two lysosomal enzymes in megakaryocytes (MK) and platelets, we examined normal human bone marrow and blood by electron microscopy and cytochemistry. Acid phosphatase (AcPase) was present in the Golgi region in the youngest recognizable MK, as well as in those with a considerable degree of cytoplasmic maturation. Heavy reaction product was usually confined to one or two Golgi-associated cisternae and coated vesicles; other Golgi cisternae were sometimes lightly reactive. In mature MK, reaction product was limited to vesicles of variable size, but smaller than alpha granules. Another lysosomal enzyme, arylsulfatase (AS), was localized in similar small vesicles in MK of all stages; it could not be demonstrated in the Golgi complex. Vesicles containing AS were also found in about 25% of platelet profiles, whereas vesicles containing AcPase were found in only about 15% of platelet profiles. The alpha-granules of all MK and platelets examined were negative for both enzymes. We conclude that the enzyme-containing vesicles in these cells constitute the lysosomes and that they are distinct from other platelet organelles. Since there was no evidence that they had participated in any digestive event, we believe that they are primary lysosomes, whose contents are secreted during platelet aggregation and the release reaction. PMID- 6277411 TI - Dexamethasone synergistically induces chemotactic peptide receptor expression in HL-60 cells. AB - We have examined the effect of physiologic concentrations of dexamethasone on the induction of differentiation of the human promyelocytic leukemia cells (HL-60) and the activity of surface receptors for the f-met-leu-phe chemotactic peptide. Dexamethasone at a concentration of 500 nM was markedly synergistic with dimethylformamide and dimethylsulfoxide, but not with trans-retinoic acid, in inducing f-met-leu-phe binding by HL-60 cells. The increase in peptide binding was due to an increase in the activity of peptide receptors rather than an increase in receptor affinity. The effect of dexamethasone was relatively specific for peptide binding as there was little effect when HL-60 differentiation was assessed by cell morphology or the ability of the cells to reduce nitroblue tetrazolium. Dexamethasone alone had no reproducible effect on HL-60 differentiation as assessed by the three criteria tested. The dexamethasone concentration required for half maximal synergism correlated with the known binding constant of the hormone for steroid receptors in HL-60 cells but the effect of dexamethasone was not blocked with the steroid blocking agents 17 alpha methyltestosterone or progesterone. PMID- 6277412 TI - Enrichment of hematopoietic progenitor cells from human bone marrow on Percoll density gradients. PMID- 6277413 TI - Cytotoxic activity in the serum of a patient with metastasizing nephroblastoma given intravenous infusions of alkyl-lysophospholipids in a phase I study. AB - In a phase I study a cytotoxic activity in the serum of a tumor patient given infusions of synthetic alkyl-lysophospholipid has been demonstrated. Serum samples collected after ALP infusions inhibited 3H-thymidine incorporation by human leukemic cells to an extent that correlated to the dose of ALP administered. Serum taken after the highest dose of ALP given (50 mg/kg body weight) led to complete cell destruction after 72 h in vitro. Whereas cells from the HL60 line were very sensitive to the serum cytotoxicity, K562 cells were much less affected. Cytotoxic activity was found to be clear from the circulation in a biphasic manner; more than 50% disappeared within 6-8 h but 20-30% was still present after 4 days. PMID- 6277414 TI - Single and multiple inert gas washout. PMID- 6277415 TI - Model analysis of inert gas mixing in the lung. PMID- 6277416 TI - Phase II of expiratory curves of respiratory and inert gases in normals and in patients with emphysema. PMID- 6277417 TI - Information content of inert gas elimination techniques. PMID- 6277418 TI - Role of diffusion-dependent gas inhomogeneity in gas exchange in the dog. PMID- 6277419 TI - Impaired oxidation of debrisoquine in patients with perhexiline neuropathy. AB - The use of perhexiline maleate as an antianginal agent is occasionally associated with side effects, particularly neuropathy and liver damage. The reason why some individuals develop these toxic reactions is not clear, though some evidence suggests that they may result from impaired oxidative metabolism, due to genetic or hepatic factors, and consequential accumulation of the drug in toxic concentrations. Drug oxidation was measured with an oxidation phenotyping procedure in 34 patients treated with perhexiline, 20 of whom had developed neuropathy and 14 of whom had not. Most of the 20 patients with neuropathy, but not the unaffected patients, showed an impaired ability to effect metabolic drug oxidation. This impairment was independent of hepatic function, concurrent drug therapy, or tobacco or alcohol consumption. The fact that the ability to oxidise several drugs is genetically controlled points to a genetic susceptibility to developing neuropathy in response to perhexiline. Routine determination of the drug oxidation phenotype might lead to safer use of perhexiline by predicting patients who may be more at risk of developing a neuropathic reaction associated with its long-term use. PMID- 6277421 TI - Importation of pathogenic Entamoeba histolytica. PMID- 6277420 TI - Human cancers and human viruses. PMID- 6277422 TI - The use of infectious bovine rhinotracheitis vaccine in a commercial veal unit: antibody response and spread of virus. PMID- 6277423 TI - Interaction of barbiturates with benzodiazepine receptors in the central nervous system. AB - The interaction of barbiturates with benzodiazepine receptors was studied in extensively washed membrane preparations from rat brain. Sedative/hypnotic and anesthetic barbiturates such as pentobarbital, and convulsant barbiturates such as DMBB, enhanced [3H]diazepam binding in a stereospecific fashion. Freeze thawing of membranes resulted in a decrease in the potency of barbiturates to enhance [3H]diazepam binding, while the maximum response to barbiturates remained unchanged. Significant differences in both the potency and maximum enhancement of [3H]diazepam binding by pentobarbital was observed among brain regions. The rank order potency of pentobarbital in different brain regions was: cerebellum greater than cortex greater than hippocampus, while the rank order efficacy of pentobarbital in these brain regions was reversed. The effects of a combination of anesthetic and/or convulsant barbiturates on [3H]diazepam binding suggested that these compounds function as partial agonists while a combination of anesthetic or convulsant barbiturates with phenobarbital suggested that latter compound antagonized the actions of both anesthetic and convulsant barbiturates. The convulsant benzodiazepine Ro-5-3663 and inosine were more potent as inhibitors of pentobarbital-enhanced than basal (non-pentobarbital enhanced) [3H]diazepam binding. Solubilization of benzodiazepine receptors with Lubrol-PX resulted in a complete loss of barbiturate enhanced [3H]diazepam binding, and greater than a 75% loss in efficacy in the remaining (insoluble receptor) tissue. These data, coupled with recent observations from this and other laboratories, suggests that the site(s) at which barbiturates act to enhance [3H]diazepam binding to benzodiazepine receptors is distinct from the site at which GABA acts to enhance [3H]diazepam binding. The phenomenon of enhanced benzodiazepine binding by barbiturates may be related to the depressant actions of the barbiturates, that is, their direct effects to increase chloride conductance. Although it is premature to assign a pharmacologic correlate to this neurochemical phenomenon, it appears that this action may be related to the anesthetic effects of the barbiturates. However, the definitive assignment of either the electrophysiologic or pharmacologic sequelae to this neurochemical action will require further investigation. PMID- 6277424 TI - Trigeminal evoked potentials: origins in the cat. AB - Short latency brain stem (far-field) and cortical (early-field) potentials were evoked by stimulation of the main nerve epidurally over the primary sensorimotor cortex. The specific generators of these volume conducted potentials were identified from both lesion and recording studies in the trigeminal lemniscal system. Far-field components I, II and III, respectively, originate from the semilunar ganglion, trigeminal lemniscus and thalamocortical radiation; whereas early near-field components P1 and N1 originate from the cortical cell layers. PMID- 6277425 TI - Distribution of alpha-adrenergic, beta-adrenergic and dopaminergic receptors in discrete hypothalamic areas of rat. PMID- 6277426 TI - Effect of naltrexone on regional brain oxygen consumption in the cat. AB - Cerebral blood flow was measured by 141Ce and 85Sr microsphere accumulation, and oxygen saturation of arterial and venous blood within 9 regions of the brain was determined by microspectrophotometric measurement in chloralose anesthetized cats. Extraction was calculated as the difference between oxygen content of arterial and venous blood. Oxygen consumption was computed as the product of blood flow and oxygen extraction. Global and regional comparisons were made between 7 anesthetized cats and 7 anesthetized cats given 1 mg/kg naltrexone HCl i.v. Naltrexone treatment reduced total brain oxygen consumption by 48% at 20 min after intravenous injection. Analysis of the anterior and posterior cortex, lenticulate nuclei, hippocampus, thalamus, hypothalamus, medulla, pons and cerebellum indicated that the oxygen consumption of the pons and hypothalamus was reduced to the greatest extent. The decreased oxygen consumption reduced blood flow in the whole brain, and the pons. The oxygen supply to consumption ratio was not altered by naltrexone except in the hypothalamus where an even greater excess of oxygen was delivered. These observations are interpreted as indicating that opiate receptor blockade is associated with reduced brain metabolism and that this response is not restricted to regions of high opiate receptor concentration. PMID- 6277428 TI - Magnitude scaling of displacement and velocity of tactile stimuli applied to the human hand. PMID- 6277427 TI - Reversal in cutaneous of Ib pathways during human voluntary contraction. PMID- 6277429 TI - Action of tetrodotoxin on pacemaker conductances in Aplysia neurons. PMID- 6277430 TI - Brain stem influences on the parasympathetic supply to the urinary bladder of the cat. AB - (1)The brain stem of anaesthetized cats has been mapped between Horsley-Clark planes APO and P8.5 with electrical stimuli of low intensity in order to determine the areas which can produce excitatory of inhibitory influences on the spontaneously contracting, sympathetically denervated, urinary bladder. (2) Two inhibitory areas were found. The first extended from P3.0 to P8.5 and at all levels was coincident with the midline raphe nuclei. The second area occurred largely 2-3 mm lateral to the midline, in the area of the nucleus reticularis pontis caudalis rostrally, and the nuclei reticularis gigantocellularis and parvocellularis caudally. Both of these areas were found to inhibit bladder contractions with threshold stimulus parameters of 20-60 microamperemeter, 400 microseconds, 20 Hz. (3)One excitatory area was found, largely 3-4 mm lateral to the midline. This area appeared large and diffuse in the lateral reticular formation. It is possible that it originated in the pontine micturition centre in the rostral pontine tegmentum. Caudally, it shifted to occupy a ventrolateral position. This excitatory area was in close approximation to, and was probably interspersed with, the lateral inhibitory area. (4) In decerebrate preparations the areas that produced excitation or inhibition had the same distribution as those found in anaesthetized animals. (5) Single shock stimulation (100 microamperemeter, 400 microseconds, 0.5 Hz repetition frequency in the excitatory area could produce firing in pelvic nerve efferents to the bladder at latencies of 60-110 ms. The amplitude of such responses was dependent on the level of intravesical pressure. (6) Stimulation of the inhibitory areas produced no evoked responses in the pelvic nerve efferents, but could inhibit reflexly evoked responses in this nerve. The similarity in the time courses of the inhibitory effects from the two areas raises the possibility that one acts via the other. PMID- 6277431 TI - Pre- and postsynaptic muscarinic receptors in surgical samples from human cerebral cortex. AB - The present study was carried out using fresh surgical material from human cerebral cortex of patients who were not medicated with atropine or other drugs known to affect the cholinergic system. The concentration of [3H]L-quinuclidinyl benzilate binding sites was 0.45 /+- 0.05 pmol/mg protein and the Kd-value of the receptor-[3H]L-QNB-complex was 0.038 /+- 0.005 nM. Agonist binding was studied by varying the concentration of carbamylcholine (10(-8) to 10(-2) M) in the presence of a constant concentration (0.2nM) of [3H]L-quinuclidinyl benzilate. The data revealed the existence of two populations of binding sites for carbamylcholine with different affinities and capacities. Presynaptic muscarinic receptors were studied in slices of the cerebral cortex, which were loaded with [3H]choline. The muscarinic antagonist, atropine (10(-6) and 10(-7) M) acting at the presynaptic muscarinic receptors enhanced the release of [3H] acetylcholine. It was also shown that muscarinic stimulation leads to elevation of cyclic GMP levels in the human cerebral cortical slices. PMID- 6277432 TI - Divalent cation, ATP-dependent [3H]leu-enkephalin uptake by synaptic vesicle fraction isolated from bovine caudate nucleus. AB - The mechanism of [3H]Leu-enkephalin uptake into synaptic vesicle fraction from bovine caudate nucleus was investigated. The simultaneous addition of 2 mM MgCl2, 2 mM CaCl2, and 2 mM ATP stimulated the uptake 22 times over the control, whereas the separate addition of these agents augmented the uptake 2.4 times at most. The addition of 2 mM MGCl2 plus 2 mM ATP and of 2 mM CaCl2 plus 2 mM ATP increased the uptake 6.6 times and 4.5 times, respectively. The cation, ATP-dependent uptake reached its half-maximal level within 10 min after the initiation of incubation at 25 degrees C, and little Leu-enkephalin was taken up at 0 degree C. The apparent Km for the uptake of [3H]Leu-enkephalin was 1.8 x 10(-7) M. Guanosine triphosphate stimulated the uptake as well as ATP, whereas CTP and ITP were only one-fourth effective of ATP. The cation, ATP-dependent uptake was inhibited by 25% and 20% in the presence of 0.1 mM colchicine and 1 microM reserpine, respectively. PMID- 6277433 TI - Analgesic effect of intrathecal morphine demonstrated in ascending nociceptive activity in the rat spinal cord an in effectiveness of caerulein and cholecystokinin octapeptide. AB - The effect of intrathecal injections of morphine and the two peptides, caerulein and cholecystokinin octapeptide (CCK-8), on the activity in ascending axons of the spinal cord evoked by electrical stimulation of primary nociceptive afferents was studied in spinal rats with decerebration. Morphine (20 microgram) depressed the spontaneous activity and the activity evoked from either A delta-or C-fibres. The co-activation by A delta-fibre stimulation of ascending axons activated by stimulation of C-fibres and the activity in ascending axons activated by stimulation of afferent A beta-fibres were not influenced by morphine. C-Fibre evoked ascending activity was also depressed by morphine (10 microgram and 5 microgram). Ascending nociceptive activity was not changed by caerulein (30 ng) and CCK-8 300 ng, but it was depressed by a subsequent injection of morphine (20 microgram). The depressant effects of morphine were abolished by an intravenous injection of concluded that: (i) an intrathecal injection of morphine selectively depressed the ascending nociceptive activity; (ii) the depression produced by morphine is an equivalent for spinal analgesia following intrathecal injection of morphine to man; and (iii) the two components of the spinal nociceptive system, the motor and the sensory path, can independently be influenced by drugs. PMID- 6277434 TI - Monosynaptic excitation of principal cells in the lateral geniculate nucleus by corticofugal fibers. PMID- 6277435 TI - Inhibitory action of the ventral noradrenergic bundle on the lateral hypothalamic neurons through alpha-noradrenergic mechanisms in the rat. AB - Electrical stimulation of the ventral noradrenergic bundle (V-NA bundle) produced 3 types of responses in lateral hypothalamic neurons: IPSPs, a polysynaptic EPSP IPSP sequence and antidromic spikes. The IPSPs were considered to be monosynaptic due to the fixed latencies seen at stimulus intensities. Iontophoretic application of an alpha-NA antagonist blocked only the presumed monosynaptic inhibition. Most of the glucose-sensitive neurons were inhibited by stimulation of the V-NA bundle. These results may account for the hyperphagia and obesity produced by selective lesions of the V-NA bundle. PMID- 6277436 TI - At the frog neuromuscular junction prostaglandin synthase inhibitors depress and PGE partially restores quantal acetylcholine release. PMID- 6277437 TI - Selective increases in the density of cerebellar beta-1-adrenergic receptors. AB - Destruction of noradrenergic neurons by 6-hydroxydopamine or chronic blockade of beta-adrenergic receptors with propranolol increased the density of beta 1 adrenergic receptors two-fold in rat cerebellum but had no effect on the density of beta 2-adrenergic receptors. The results suggest that even through beta 1 receptors comprise only 5-10% of the total number of Beta-adrenergic receptors in the cerebellum they are the receptors specifically innervated by noradrenergic neurons and they may thus be the physiologically important receptors. PMID- 6277438 TI - The responses of dorsal horn neurones to controlled mechanical stimulation of the hindlimb skin in the rabbit. AB - Seven types of neurones responding to mechanical stimulation of the hindlimb skin were identified in the dorsal horn of the spinal cord of the rabbit. Of the 218 units examined, 33% were activated by movement of hairs--H cells; 25% were excited by stimulation of Type I domes--on/off cells; 15.6% were spontaneously active and were excited by vertical displacement of the skin and lateral skin stretch--SPE (spontaneous excited) cells; 4.4% were spontaneously active and were inhibited by mechanical stimulation of the skin--SPI (spontaneous inhibited) cells; 5.2% were spontaneously active and responded to application of pressure- SPEP (spontaneous excited pressure) cells; 3% were spontaneously active and were excited by movement of joints--SPEJ (spontaneous excited joint) and 13.8% of neurones responded to hair movement and also to some other forms of mechanical stimulus--CM (combined mechanosensitive) cells. Of the 218 units examined, 133 were located in lamina IV and 85 in lamina V. A quantitative analysis of some of the characteristics of the on/off, SPE and SPI cells is carried out and a model of the dorsal horn system activated by Type I and Type II receptor units is proposed. PMID- 6277439 TI - Behavior during hippocampal microinfusions. I. Norepinephrine and diversive exploration. AB - Adult male Sprague-Dawley rats were surgically implanted with guide cannulas in the anterodorsal hippocampal formation. After recovery from surgery they were administered bilateral infusions into the dentate gyrus of D,L-norepinephrone (NE) at the rate of 0.025 microliter/min throughout a 40-min session in a holeboard/activity apparatus. The computerized holeboard system measured the animals' locomotor activity, stimulus responsivity, and response to novelty, and permitted the reconstruction and analysis of their sequential patterns of movement. The NE infusions failed to affect overall locomotion or holepoking, although group means were slightly elevated. Rearing frequency and duration were significantly increased, as were the number of different holes poked per 5-min epoch and the amount of time in and number of entries into the center region of the holeboard. The novel object reaction of NE-infused rats, measured as the increase in poke duration into holes with novel stimuli, was indistinguishable from that of saline-infused rats. The NE-infused rats exhibited more varied, widespread spatial distributions of sequential patterns of locomotor activity. A dose of the beta-adrenergic antagonist propranolol that was minimally effective when infused by itself blocked all of NE's affects when co-infused in the same solution. Infusions of the noradrenergic releasing agent tyramine mimicked NE's actions, whereas infusions of NE's relatively inactive stereoisomer D-NE or infusions into the overlying lateral ventrical failed to do so. Histological examination of dye-infused brains and microspectrofluorimetry of NE-infused brains treated using the Falck-Hillarp technique for the formaldehyde-induced fluorescence of monoamines indicated that spread of the infusate was confined to the dentate gyrus of the anterodorsal hippocampal formation. The behavioral profile of the NE-infused rats suggests a role for the noradrenergic input to the hippocampal formation in spontaneous environmental reconnaissance and the diversification of stimulus sampling-'diversive' exploration, as opposed to the inspection or 'specific' exploration of unfamiliar stimuli. PMID- 6277440 TI - [Blood levels of hormones, prostaglandins, and cAMP in crew members of the space flight "Soyuz 28" (author's transl)]. PMID- 6277441 TI - Chloroplast deoxyribonucleic acid isolation: purity achieved without nuclease digestion. PMID- 6277442 TI - Lobar pneumonia associated with adenovirus type 7. PMID- 6277443 TI - "Labrador lung": an unusual mixed dust pneumoconiosis. AB - Forty-eight workers involved for various periods from 1962 in mining and milling iron-containing rock in the Carol Lake area of western Labrador were found in late 1974 and early 1975 to have pneumoconiosis. Their annual chest roentgenograms demonstrated irregular nodular densities, and several revealed mild hilar lymphadenopathy or pleural thickening. All 48 workers were asymptomatic and had normal pulmonary function; their average age was 38 years. At work they had been exposed to high levels of dust containing iron, silica and anthophyllite, a type of asbestos. Lung biopsy specimens from 13 of them showed widespread focal fibrosis, large amounts of hemosiderin and silica and many ferruginous bodies; one ferruginous body was identified as having a core of anthophyllite. A granulomatous reaction was seen in two biopsy specimens and typical silicotic nodules were found in two others. The relatively brief exposure, especially in ones case (11 months), and the short latent interval before the development of the pneumoconiosis are reasons for concern for the health of iron ore workers. PMID- 6277444 TI - Prenatal prediction of alpha-thalassemia phenotype by endonuclease mapping of parental DNA. PMID- 6277445 TI - Intractable pain with breast cancer. AB - This study examines retrospectively the cause, clinical features, natural history and results of treatment of intractable pain associated with breast cancer in 210 patients. The three chief types of pain were that due to skeletal metastases or brachial plexus neuropathy and pain of psychogenic origin. Onset at the time of cancer diagnosis characterized the psychogenic pain, whereas pain from metastases first occurred after a median latency of 3.7 years. Treatment was custom-tailored to the specific patient and pain problem, with several factors taken into account. The onset of intractable pain due to metastatic disease indicated a short survival (median, 9 months). PMID- 6277446 TI - Postirradiation sarcoma (malignant fibrous histiocytoma) following cervix cancer. AB - A case of postirradiation sarcoma is described. The tumor, a malignant fibrous histiocytoma, occurred in the radiation field 11 years following postoperative external beam radiation therapy (7000 rad) for carcinoma of the cervix. Reports of postirradiation malignant fibrous histiocytoma are rare, and the occurrence of this neoplasm following treatment of cervix cancer has not previously been described. The literature concerning postirradiation bone and soft tissue sarcomas is briefly reviewed, with special attention to malignant fibrous histiocytomas. PMID- 6277447 TI - Hepatocellular carcinoma arising in noncirrhotic and highly cirrhotic livers: a comparative study of histopathology and frequency of hepatitis B markers. AB - Hepatocellular carcinoma (HCC) associated with cirrhosis and HCC developing in a noncirrhotic liver may have differing pathogeneses. To study this possibility, 425 autopsied cases of HCC were investigated. Of these, 45 livers were not cirrhotic, 50 were highly cirrhotic (liver weight less than 99 g), and the remaining 331 were cirrhotic but not so highly. The average age was significantly older in the highly cirrhotic group, suggesting a longer premalignant period of chronic liver disease. The liver weight in the noncirrhotic group was about 3.5 times that in the highly cirrhotic group. Hepatitis B surface antigen was positive in serum in only 9.3% and in liver tissue in 10% in the noncirrhotic cases, the positivity rate being much lower compared with other groups (P less than 0.005--0.01), yet antibody to HB core was positive in 90%. The antibody titers were low, however, indicating that these noncirrhotic patients had in the past had HB virus (HBV) infection with no residual chronic B hepatitis. Analysis of the grades of anaplasia of cancer tissue demonstrated an inverse correlation between the degree of fibrosis and grade of anaplasia, i.e., the more advanced the fibrosis, the less anaplastic the cancer. These data suggest that HCC arising in highly cirrhotic liver and in noncirrhotic livers have different pathogenetic backgrounds, and that HBV infection, even though transient, has a certain role in hepatocarcinogenesis. The generally held conjecture that HCC in a noncirrhotic liver is caused by nonviral carcinogens and HCC arising on the ground of cirrhosis is due to HBV seems untenable in such a simple concept. PMID- 6277448 TI - Enzyme pathology and the histologic categorization of human lung tumors: the continuum of quantitative biochemical indices of neoplasticity. AB - The purpose of the present enzymic and histologic analysis of pulmonary samples from 39 subjects was to discern a common, meaningful pattern which may underlie the biochemical heterogeneity of lung neoplasms. The distribution among the different tumors of thymidine kinase, uridine kinase, phosphoserine phosphatase, hexokinase and adenylate kinase was found to correlate with each other. By averaging their standardized units (normal lung = 0) an enzymic index of neoplasticity was calculated for each tumor and used (in increasing order) to rank all 39. The index, showing a significant positive correlation with mitotic frequency, encompassed a continuous 100-fold range. Poorly differentiated carcinomas ranked high while neoplasms with better differentiation and prognosis placed in the lower half of the range. The results indicate that enzymes showing coordinated variations over a broad spectrum of tumors could contribute objective criteria to the rating of any individual tumor against a continuous, quantitative scale of neoplasticity. PMID- 6277449 TI - Vulvar intraepithelial neoplasia: correlation of nuclear DNA content and the presence of a human papilloma virus (HPV) structural antigen. AB - Immunoperoxidase localization of a human papilloma virus structural antigen (HPV) was attempted in 68 intraepithelial lesions of the vulva, 39 of which were analyzed for nuclear DNA content by microspectrophotometry. Overall, 5.9% (4/68) stained positive for HPV. Ninety percent (35/39) of the cases tested were aneuploid, and, of these, 2.8% (1/35) stained positive for HPV. In contrast, 50% (2/4) of the polyploid lesions were positive. Hence DNA microspectrophotometry and immunoperoxidase localization of HPV are useful coparameters for distinguishing wart virus infection (condylomata) from vulvar intraepithelial neoplasia. HPV is detected infrequently within aneuploid lesions, in keeping with the concept that epithelial maturation is required for virion assembly. Whether the HPV genome exists in a nonreplicative state within the aneuploid cell population is unknown. PMID- 6277450 TI - The significance of bone and cartilage formation in malignant fibrous histiocytoma of soft tissue. AB - Malignant fibrous histiocytoma of soft tissues (MFH), a mesenchymal tumor of varied morphologic patterns and cell types, sometimes contains bone and cartilage. Such bony and cartilaginous elements in a pleomorphic MFH may pose a difficult diagnostic challenge. An MFH with bone and cartilage can be distinguished from extraosseous osteogenic sarcoma and chondrosarcoma on the basis of qualitative and quantitative features of the osseous and chondroid elements. Five cases of soft tissue MFH containing bone and cartilage reported here showed: 1) that the bony and cartilaginous elements are mostly metaplastic and tend to be in the pseudo-capsule or fibrous septa of the tumor; and 2) that the bony elements may show a zoning pattern with peripheral maturation similar to myositis ossificans. In a single case, there was a small amount of "tumor osteoid." However, this was focal and lacked the diffuse and finely divided or ribbon-like pattern generally associated with osteogenic sarcoma. The probability that MFH with bone and cartilage is less aggressive than soft tissue osteogenic sarcoma or chondrosarcoma underscores the importance of accurate histologic diagnosis of these tumors. PMID- 6277451 TI - Myoepithelioma of salivary glands: report of 23 cases. AB - Twenty-three neoplasms arising in major and minor salivary glands have been found to be composed of two types of myoepithelial cells--the plasmacytoid and spindle cell form. Twelve of these were located within the parotid gland and the submandibular gland and intraoral minor salivary glands accounted for five each. A solitary example was found within the upper lip. Sixteen (70%) of the cases were of the spindle-cell type, four (17%) were composed of plasmacytoid cells, and three cases (13%) contained both cell forms in approximately equal quantity. Differences in behavior, recurrence rate, frequency, age of the patient, and location were not related to cell type. Ultrastructural analysis of selected cases showed myofilament aggregation patterns to vary between the two subtypes. Previously published cases confirmed by electron microscopy have been reviewed and compared with the 23 cases reported. Conservative surgical management is curative. Follow-up information on 16 cases is presented. PMID- 6277452 TI - Mesoblastic nephroma in an adult patient: recurrence 21 years after removal of the primary lesion. AB - A histologically benign mesoblastic nephroma was diagnosed in a 19-year-old girl. The tumor was resected but recurred, invading the liver 21 years after the removal of the primary lesion. The recurrent tumor was composed of fibroblastic cells surrounded by collagen and elastin-rich interstitial material. Some tumor cells also contained fibrillar bodies, most likely representing extracellular material packaged for secretion. PMID- 6277453 TI - Variability of histologic pattern in recurrent soft tissue sarcomas originally diagnosed as liposarcoma. AB - The histopathologic classification of liposarcoma and its clinicopathologic significance are well delineated in previous studies, but the literature is inexplicably mute on recurrent liposarcomas. Supposition and experience may have led to the assumption that the microscopic features remained static over a period of years and multiple recurrences. Thirteen sarcomas originally classified as liposarcoma were reviewed to assess patterns of histologic change with recurrence. A total of 52 recurrences was studied. In 7 of the 13 cases, multiple morphologic sarcomatous patterns were identified, either as a change with recurrence or as multiple discrete patterns within a tumor. All changes except in one case occurred before irradiation or chemotherapy. The variations included patterns such as malignant fibrous histiocytoma, hemangiopericytoma, malignant schwannoma, and unclassified spindle-cell sarcoma as well as the various liposarcoma subtypes. This variability of patterns can cause significant diagnostic difficulties and can cause one to question the validity of using monomorphic diagnoses in all cases. In addition, the polymorphic differentiation of these tumors supports the concept that soft tissue sarcomas are derived from a pluripotential mesenchymal stem cell. PMID- 6277454 TI - Possible enhancement of vincristine neuropathy by VP-16. AB - Eleven consecutive patients with small cell carcinoma of the lung treated intensively with a combination chemotherapy regimen cyclophosphamide, Adriamycin, VP-16, and vincristine experienced peripheral neuropathy. Four of the 11 patients had severe (Grade III) neuropathy, leaving them virtually bedridden; the remaining seven patients had mild to moderate neuropathy. Only 8 of the 14 patients treated less intensively with vincristine and VP-16 had Grade I and II neuropathy. Predisposing factors for severe neuropathy included advanced age and preexisting peripheral neuropathy. All patients' performance status declined and all lost much weight before developing neuropathy. From accumulated treatment experience of small cell carcinoma with vincristine alone or with vincristine, Adriamycin, cyclophosphamide combinations, the authors feel that the severe neuropathy observed was due to an interaction of vincristine and VP-16. This conclusion is supported by electron microscopic observation of electronopaque granular degeneration of the myelin lamellae of affected nerve fibers. PMID- 6277455 TI - Origin of bronchioloalveolar carcinoma and peripheral bronchial adenocarcinoma. AB - The morphologic distribution of newly synthesized glycoproteins in a bronchioloalveolar carcinoma and in a peripheral bronchial adenocarcinoma with areas of bronchioloalveolar differentiation was determined and compared with bronchiolar epithelium and type II alveolar cells. Tissues were maintained in vitro, and autoradiography was used to reveal locations of glycoproteins metabolically labelled with 3H-glucosamine. Both tumors displayed glycoprotein secretory activity as most incorporated radioactivity was rapidly released from cells. Silver grains were associated with cytoplasmic granules and mucinlike secretions in the bronchioloaveolar carcinoma and granules plus unidentified secretions in the bronchial adenocarcinoma. Clara cells exhibited an association of newly synthesized glycoproteins with cytoplasmic granules similar to those found in the tumors. In bronchiolar ciliated epithelium, as in bronchial ciliated cells and central bronchogenic adenocarcinomas, glycoproteins accumulated over apical cell surfaces and were not rapidly secreted. Type II cells incorporated label into secreted contents of cytosomal inclusions and unidentified cytoplasmic material. The observations suggest that a bronchioloalveolar carcinoma and a peripheral bronchogenic adenocarcinoma are distinct from common bronchogenic adenocarcinomas and are derived from secretory cells that resemble bronchiolar Clara cells. PMID- 6277456 TI - Ductuloinsular tumors of the pancreas: a light, electron microscopic and immunohistochemical study. AB - Most pancreatic tumors are of a single cell type and are identified delta as duct, acinar, or islet cell neoplasms. The authors report on three examples with both duct and endocrine characteristics as seen by light microscopy; two with further confirmation of endocrine differentiation by electron microscopy; and one by immunocytochemistry. Mixed differentiation of this sort can be understood by reference to the embryonic pancreas, which develops from the small intestine and forms ducts, intercalated ducts, acini, and islets, with their different cell types. The merging and intermingling of different cell prevents the identification of one specific cell of origin. It also suggests that the neoplastic process here may not be a clonal proliferation from a single cell mutation as this is generally understood. Alternative explanations are briefly mentioned. PMID- 6277457 TI - Multicentric breast carcinomas: clinical-radiographic-pathologic whole organ studies and 10-year survival. AB - Selection of breast cancer patients with microscopically limited disease but with excess mortality or women with regional disease and lessened mortality has remained an unsolved and critical challenge. The many usual histologic features such as tumor size, stage of disease or tumor differentiations were found reduced to lessened significant prognostic factors by the presence of multicentricity of tumors and multiplicity of histologic types of carcinoma. This observation was noted upon review of 161 clinical, radiographic and histopathologic whole breast studies on 156 patients with follow-up from 11-15 years. Epidemiologic, clinical, and radiographic data had been compiled prospectively prior to diagnosis. Women with single site and single type of carcinoma had a better prognosis (2.5% mortality per year) than those with multiple sites and multipe types (15% mortality per year) even though the stage of the disease may be similar. When the latter groups contained a scirrhous type duct carcinoma, the annual mortality rate approached 25%. PMID- 6277458 TI - Localization of occult breast lesions. AB - Localizing nonpalpable suspicious lesions seen by mammography is a challenge to the surgeon. Using a previously reported technique we report our experience in 66 needle localization procedures for nonpalpable lesions of which fourteen were carcinoma. We emphasize the ease and accuracy of this technique with minimal loss of breast substance. PMID- 6277459 TI - Radiotherapy as an adjuvant in the treatment of carcinoma of the breast. AB - During the period from 1950-1974, 510 patients with a diagnosis of invasive ductal carcinoma treated with the standard Halsted radical mastectomy were reviewed retrospectively from the records of the Tumor Clinic of The Memorial Hospital, Danville, Virginia. The primary aim of the study was to determine whether postoperative adjuvant radiation therapy (POART) as delivered in a community hospital setting contributed to better cancer control and effectively influenced five and ten year survival rates. The results of the review indicated that the use of POART whether from an orthovoltage or cobalt source did not differentially influence the patient's disease outcome. The use of POART did not improve the five- and ten-year survival rates regardless of lesion location in patients with Stages I, II, and III disease with positive lymph nodes. POART did cause a significant decline in five- and ten-year survival rates of patients with Stage II and III disease with negative nodes when compared to the survival of patients with the same stage of disease that did not receive POART, i.e., 75% versus 94% five-year difference, and a 65% versus 85% ten-year difference. These results suggest that radiotherapy may have a detrimental effect on immunocompetence and thus alter survival rates in certain circumstances. Therefore, any clinical investigation measuring the results of various therapeutic modalities singularly or in combination should be based on pathologic staging rather than clinical staging so that these factors can be adequately evaluated. PMID- 6277460 TI - The natural history of neoplastic development: the relation of experimental models to human cancer. AB - The natural history of the development of neoplasia in experimental systems may be separated into at least three different stages, those of initiation, promotion and progression. Evidence for such distinct stages has been demonstrated in at least half a dozen different experimental systems. The stage of promotion is that stage most easily modulated. The actions of promoting agents are reversible and only effective above certain threshold levels of the promoting agent. This is in contrast to the stage of initiation, which can be induced at any dose of the carcinogenic agent in an irreversible manner. Agents exhibiting both initiating and promoting activities are termed complete carcinogens while incomplete carcinogens are those capable only of initiation. Promoting agents do not initiate but may promote cells initiated by ambient environmental means, giving the appearance of complete carcinogens in standard bioassay procedures. The stages of initiation and promotion have been extensively studied in skin and liver carcinogenesis and show almost identical characteristics. A variety of promoting agents in the human environment have been demonstrated both by experimental and epidemiologic methodologies. The importance of an understanding of tumor promotion in relation to the prevention of human cancer is emphasized. PMID- 6277461 TI - Brain stem gliomas: an autopsy study of 25 cases. AB - The study analyzed the extent of tumor at autopsy in 25 patients with a brainstem glioma. Primary tumor in the pons comprised the majority of cases, followed by medulla and midbrain. Histologically, 48% of the tumors were glioblastoma multiforme. The tumor spread was found to be dependent on the site of origin and the grade. Pontine tumors involved the adjacent structures more extensively than the tumors of midbrain and medulla. Contiguous cephalad and caudad involvement was frequent with Grade III and Grade IV tumors. The extent of meningeal involvement was limited to those surrounding the main tumor mass. It is concluded that in the treatment of brainstem gliomas, irradiation through fields limited to the brainstem, adjoining temporal lobes, cerebellum, and proximal spinal cord would be adequate. PMID- 6277462 TI - Long-term follow-up in radiation therapy of carcinoma of the vagina. AB - A retrospective analysis of 134 patients with histologically confirmed carcinoma of the vagina is reported. Actuarial disease free 5-year survival was: Stage 0 (15 patients), 90%; I (39 patients), 90%; IIA (39 patients), 58%; IIB (21 patients), 32%; III (12 patients), 40%; IV (8 patients), 0%. Fifteen patients had carcinoma in situ; of these, 14 were controlled with interstitial or intracavitary therapy. Of 39 patients with Stage I carcinoma, 37 (95%) showed no evidence of vaginal or pelvic recurrence. Most of them received interstitial or intracavitary therapy or both; the addition of external beam irradiation did not significantly increase survival or tumor control. In Stage IIA (paravaginal extension) 22 of 34 (64.7%) patients were controlled with a combination of brachytherapy and external beam irradiation; only two of five (40%) treated with brachytherapy alone exhibited tumor control in the pelvis. Eight of 12 Stage III tumors were controlled in the pelvis. Only two of eight patients with Stage IV had no recurrence in the pelvis even with relatively high doses of irradiation. The dose of irradiation delivered to the primary tumor or the parametrial extension was critical in achieving successful results. The incidence of complications (9.7%) is correlated with the stage of the tumor and type of treatment given. PMID- 6277463 TI - Presence of cellular retinol and retinoic acid-binding proteins in epidermoid carcinoma of the oral cavity and oropharynx. AB - Epidermoid carcinomas of the oral cavity and oropharynx from six patients were examined for the presence and amount of cellular retinol (CRBP) and cellular retinoic acid-binding (CRABP) proteins. In all cases adjacent, grossly normal tissue was similarly examined. For each example CRBP levels were significantly higher in tumor tissue compared to adjacent tissue. In four cases CRABP was significantly higher. This is of interest because retinol, retinoic acid and their analogs have been shown to inhibit the development of various epithelial tumors, and this inhibition is possibly mediated by these binding proteins. PMID- 6277465 TI - Identification of lymphocyte subpopulations in human breast cancer tissue and its significance: an immunoperoxidase study with anti-human T- and B-cell sera. AB - Subpopulations of the infiltrating lymphocytes in breast cancer tissue from 31 patients were identified by indirect immunoperoxidase technique with antihuman T- and B-cell sera. In all noncancerous lesions examined (seven cases), B-cells were predominant and T-cells were scarcely found. In contrast, T-cells were predominant in breast cancer tissues (17 in 21 cases). T-cells tended to contact closely with cancer cells or cancer cell nests and accumulated around and in the walls of venules draining the cancer, while B-cells tended to cluster focally apart from cancer cell nests. T-cell infiltration was scanty in scirrhus carcinoma, whereas it was ample in infiltrating papillotubular carcinoma which had a better prognosis. There was a significant reverse correlation between the intensity of the T-cell infiltration and the clinical stages. The intensity of the T cell infiltration was significantly high in patients without lymph node metastasis. These facts suggest the possibility that the infiltrating T-cells in cancer tissue represent host resistance against cancer and that the intensity of the T cell infiltration correlates with the clinical prognosis of the breast cancer patients. PMID- 6277464 TI - Abnormal bone and parathyroid histology in carcinoma patients with pseudohyperparathyroidism. AB - Postmortem bone and parathyroid gland histology in nine hypercalcemic cancer patients without bone metastases was compared to bone and parathyroid histology in ten normocalcemia patients. Parameters of parathyroid function, including serum immunoreactive parathyroid hormone, acid base status, serum phosphate, and nephrogenous cyclic AMP were measured in the hypercalcemic group and compared to normals and to patients with primary hyperparathyroidism. Bone histology in all nine hypercalcemic cancer patients showed increased osteoclastic bone resorption and increased fibrous connective tissue in the bone marrow. Parathyroid glands were of normal size in all nine patients but contained little or no fat, one criterion of parathyroid hyperplasia. In the normocalcemic cancer patients only 2/10 had minimally increased bone resorption while 7/10 had decreased or absent stromal fat in the parathyroid glands. Despite the hyperplastic appearance of the parathyroid glands, serum biochemical parameters in the hypercalcemic cancer patients indicate a state of suppressed parathyroid function suggesting that the osteoclastic bone resorption is related to a humoral substance elaborated by the tumors which is distinct from parathyroid hormone. PMID- 6277466 TI - Breast carcinoma in a 46, XX true hermaphrodite. AB - Muscinous breast carcinoma occurred in a 67-year-old true hermaphrodite who had been reared as a male. The peripheral blood karyotype was 46, XX. A left ovary and right ovotestis were intra-abdominal. Other tumors included an ovarian thecoma and an intestinal leiomyoma. Two previously reported cases of breast cancer in true hermaphrodites are cited. PMID- 6277467 TI - Abdominal computed tomography in small cell lung cancer: assessment of extent of disease and response to therapy. AB - Abdominal computed tomography (CT) was performed as part of the initial staging evaluation in 77 patients with small cell carcinoma (SCC) of the lung. CT scans revealed mass lesions in 26 patients (34%). Abnormalities were confined to the liver in 15 patients and to retroperitoneal structures (lymph nodes, adrenal glands, psoas muscle region masses) in eight, and occurred in both areas in three. However, only three of 29 patients otherwise staged as having limited disease (confined to one hemithorax and regional nodes) had evidence of abdominal metastases on CT scan. Most (23/26) positive studies were in patients already known to have more extensive tumor dissemination. In 71 patients with pathologic confirmation of liver status, CT had a sensitivity of 63%, specificity of 91%, and overall accuracy of 85% in assessing the liver. Comparison of radionuclide liver scan findings with hepatic biopsies gave similar results. During therapy, 65 follow-up CT scans were obtained in 46 patients. Scan abnormalities improved or disappeared in 11/12 cases with tumor response documented in other ways, appeared or worsened in 5/13 cases of tumor progression that was diagnosed by other tests, and only rarely (2/65 scans) improved at the time of documented tumor progression, or vice versa. In only three patients, however, did CT scan provide the sole site of evaluable disease during treatment or detect either the only area of residual disease in a patient in otherwise complete remission or the initial evidence of tumor progression. Although abdominal CT scans in SCC can demonstrate metastatic dissemination not evaluable by other means, they provide relatively little therapeutically relevant information beyond that obtained with standard staging procedures. PMID- 6277468 TI - Minute hepatocellular carcinoma without appreciable change in size for seven years: a case report. AB - A small hepatocellular carcinoma found and followed in a Japanese man is described. He died of diabetic nephropathy and bleeding esophageal varices seven years after surgical biopsy of the liver that revealed hepatocellular carcinoma. Clinical and autopsy findings demonstrated no appreciable change in tumor size during the seven-year period during which no specific treatment was given for cancer. PMID- 6277469 TI - Chromosome #14 markers in two Epstein-Barr virus (EBV)-transformed lymphoblastoid cell lines of normal origin differ from the Burkitt lymphoma (BL)-associated 14q+ marker. AB - Two among ten Epstein-Barr Virus (EBV)-transformed lymphoblastoid cell lines contained a 14q+ marker in a low frequency of the cells (2 and 9%). By means of "mesome-prosome" analysis of the G-band patterns of these markers; it was established that the additional chromosome segments of these two 14q+ markers came from chromosomes #3 and #5, respectively, and not from chromosome #8 as in the 14q+ marker of Burkitt lymphoma. Chromosome #8 was not involved at all in any changes in the ten lymphoblastoid cell lines studied. PMID- 6277470 TI - Poly(ADP-ribose) synthesis in human cervical cancer cell-diagnostic cytological usefulness. AB - Poly(ADP-ribose) synthetase activity in human cervical cancer cell nuclei was found to be approximately twice that of normal cervical cell nuclei. Using antipoly(ADP-ribose) antibody, frozen section of normal tissue were stained by indirect-immunofluorescence and it was demonstrated that the fluorescence of nuclei in squamous cells decreased during maturation. Correspondingly, superficial cells appearing in smear specimens had less nuclear fluorescence compared with parabasal cells in the same smear. However, cancer cells in cervical smears had marked nuclear fluorescence using immunostaining and were easily distinguished from normal cells. PMID- 6277471 TI - Carcinogenicity in rats of the mutagenic compounds 1-nitropyrene and 3 nitrofluoranthene. AB - 1-Nitropyrene and 3-nitrofluoranthene are present in diesel exhaust, in pollutants in air, and were also present in certain xerographic toners and copies. Their carcinogenicities were studied in male F344/DuCrj rats by subcutaneous injection. Sarcomas, mainly malignant fibrous histiocytomas at the site of injection were induced in 8 to 17 (47%) rats by 1-nitropyrene and in 4 of 10 (40%) rats by 3-nitrofluoranthene. Some tumors were serially transplantable in the same strain of rats. PMID- 6277472 TI - Nuclear catalyzed antibiotic free radical formation. AB - Nuclei isolated from rat liver, heart, and kidney catalyze oxygen consumption in the presence of reduced pyridine nucleotide (NADPH) and quinone or quinone-imine antibiotics such as Adriamycin, daunorubicin, actinomycin D, mitomycin C, and streptonigrin. The Km and Vmax values for NADPH were 2.4 x 10(-3) M and 3 x 10( 8) mol O2 per min per mg protein and Km values for the antibiotics ranged from 1.4 x 10(-4) M to 5.9 x 10(-6) M. Metabolism of the anthracycline antibiotics, i.e., reductive glycosidase reaction, occurs in reaction mixtures after all oxygen is consumed. During the reaction, free-radical species of Adriamycin and daunorubicin are detectable by electron paramagnetic reasonance spectrometry. These observations indicate that some cytotoxic antibiotics can be activated to a free-radical state at the site where damage to nuclear DNA may result. PMID- 6277473 TI - Specificity of the control of tumor formation by the blastocyst. AB - An assay to determine the mechanism of regulation of embryonal carcinoma cells by the blastocyst, which is based on a comparison of tumors produced when the cancer cells are cloned alone or after incorporation into blastocysts, was refined by labeling embryonal carcinoma cells with fluorescent microspheres and by following their fate after injection into the blastocysts. Through the use of the new techniques, it was observed that cells of one line of nullipotent embryonal carcinoma were controlled at the 50% level, those from another were not controlled, and those from a multipotent but undifferentiated line were controlled in almost absolute fashion. Single Sarcoma 180 of L1210 leukemia cells were not controlled when injected into the blastocele, but C1300 neuroblastoma cells were partially controlled. None of these tumors have a normal cellular counterpart in the blastocyst, as does embryonal carcinoma, but neurulation follows blastulation by only a few days, so that the neuroblastoma cells may be regulated at that time. Parietal yolk sac carcinoma cells, which have a counterpart in the late blastocyst, were not controlled. On the basis of these data, it is postulated that, if one embryonic field can regulate its closely related cancer, then there may be an embryonic field capable of regulating each carcinoma. PMID- 6277474 TI - Collateral sensitivity of 6-mercaptopurine-resistant sublines of P388 and L1210 leukemia to the new purine antagonists, 5-carbamoyl-1H-imidazol-4-yl piperonylate and 4-carbamoylimidazolium 5-olate. AB - Two new purine antagonists, 5-carbamoyl-1H-imidazol-4-yl piperonylate (SL-1250) and 4-carbamoylimidazolium 5-olate (SM-108), were investigated for their antitumor activities against 6-mercaptopurine (6-MP)-resistant sublines of P388 and L1210 leukemia. It was found that both resistant sublines exhibited collateral sensitivity instead of cross-resistance to these new antipurine drugs. Since more potent cytotoxic activities of these drugs against 6-MP-resistant cells were observed even in vivo cell culture systems, this collateral sensitivity was proved on a cellular basis. Biochemical studies revealed that 6 MP-resistant sublines of both P388 and L1210 leukemia are deficient in hypoxanthine-guanine phosphoribosyltransferase activity. In these cells, not only the activation of 6-MP to its nucleotide but also the synthesis of guanosine 5' monophosphate via the salvage pathway seems to be severely restricted. However, SL-1250 and SM-108 can be activated to their nucleotide even in these 6-MP resistant cells because the activation of these compounds is proceeded by adenine phosphoribosyltransferase. In conclusion, suppression of de novo purine synthesis with SL-1250 and SM-108 seems to be a very efficient means of killing these 6-MP resistant cells, which lack a salvage pathway for guanosine 5'-monophosphate. PMID- 6277475 TI - Presence of 1,25-dihydroxyvitamin D3 receptors in established human cancer cell lines in culture. AB - Receptors for 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3] have been described in several human breast cancer cell lines and more recently in human melanoma. The presence of 1,25-(OH)2D3 receptor (1,25-DR) in two cultured breast cancer cell lines was associated with receptors for calcitonin, another hormone thought to have effects on calcium handling. Therefore, it seemed important to examine a range of established human cancer cell lines for the presence of receptors for 1,25-(OH)2D3 and calcitonin. Thirty-three cancer cell lines were examined. 1,25 DR was found to be present in 23 lines, while calcitonin receptors were not detected in any of them. The 1,25-DR from several cell lines sedimented at about 3.5S in sucrose density gradients, had the appropriate specificity for vitamin D metabolites, had Kds of 0.8 to 2.2 x 10(-11) M, and had receptor concentrations of 12 to 99 fmol/mg protein. Ten malignant melanoma and nine colonic carcinoma lines constituted the largest groups of carcinoma cell lines, and seven and eight, respectively, of these were 1,25-DR positive. The high frequency of 1,25 DR positivity in the cultured colonic carcinoma cells is quite different from the low frequency of 1,25-DR in primary colonic carcinomas. It was also interesting that both of two cell lines derived from patients who had had both bone metastases and malignant hypercalcemia were 1,25-DR positive. These various cell lines may provide useful models for the examination of 1,25-(OH)2D3 action in vitro. PMID- 6277476 TI - Pericellular matrix and cell surface glycoproteins of virus-transformed mouse epithelial cells. AB - Mouse embryo Mus musculus castaneous epithelial cells, transformed with Moloney murine sarcoma virus (MSV) or with ecotropic murine leukemia virus (MuLV), were analyzed for production of pericellular matrix glycoproteins. The nontransformed, MSV-transformed, and MuLV-transformed cells produced fibronectin, laminin, type I collagen, and small amounts of type III collagen when studied by immunofluorescence using specific antibodies. The virus-transformed epithelial cells produced enhanced amounts of fibronectin into their growth media. Nontransformed M. musculus castaneous epithelial cells mainly produced type I collagen, as shown by metabolic labeling and polypeptide analysis. A significant increase in the glycoprotein production was seen by the MuLV-transformed cells, whereas small changes in the collagen production were apparent after MSV transformation. MuLV-transformed cells produced increased amounts of type I collagen and also some collagenous polypeptides that comigrated with procollagen type IV chains. The ratio of the procollagen type I chains deposited in the matrix was altered in transformed cells. Radioactive surface labeling of the cells revealed changes of the high-molecular-weight glycoproteins in both the MSV and the MuLV-transformed cells. Unlike virus-transformed fibroblastic cells, these transformed epithelial cells deposited and retained connective tissue glycoproteins in their pericellular matrices. The results indicate that viral transformation modulates the pericellular matrix and surface glycoproteins of cultured mouse epithelial cels. The ability of virus-transformed epithelial cells to deposit pericellular matrices is a major difference between them and virus transformed fibroblastic cells. PMID- 6277477 TI - Mammary tumor virus DNA as a marker for genotypic variance within hormone responsive GR mouse mammary tumors. AB - Mammary tumors of GR mice acquire extra mammary tumor virus (MMTV) DNA information within their DNA during tumor growth and development. These extra MMTV genes have been used by us as genotypic markers to investigate the heterogeneity of GR mammary tumors and their loss of hormone dependence during serial transplantation. Our studies reveal that the various subpopulations of cells within individual GR mammary tumors are characterized by differences in number and location of acquired extra MMTV DNA fragments. Losses of certain of these extra MMTV DNA fragments occur when mammary tumors become hormone independent, indicating a loss of hormone-dependent cells. The study of MMTV DNA markers also reveals that low levels of autonomous cells are already present in some hormone-dependent mammary tumors at an early stage of their development. The genotypic analysis strongly indicates that mammary tumor progression is not due to phenotypic adaptation but to clonal selection of the more aggressive sublines. PMID- 6277478 TI - Comparison of benzo(a)pyrene metabolism in bronchus, esophagus, colon, and duodenum from the same individual. AB - The metabolism of benzo(a)pyrene has been investigated in cultured normal human bronchus, colon, duodenum, and esophagus obtained from the same patient. The highest total metabolism was found in bronchus and duodenum, while the highest mean binding level was observed in the bronchus followed, in order, by the esophagus, duodenum, and transverse colon. A 30-fold interindividual variation in the binding level was found in each of the four organs studied, and a positive correlation between the binding levels in bronchus, colon, and duodenum was found. In human bronchus, a positive correlation was found between level of binding of benzo(a)pyrene to DNA and the amount of both benzo(a)pyrene 7,8-diol and the combined group of 3-hydroxybenzo(a)pyrene, benzo(a)pyrene 9,10-diol, and water-soluble metabolites. A significantly higher relative amount of benzo(a)pyrene tetrols and benzo(a)pyrene 9,10-diol was formed by human bronchus compared to the gastrointestinal tissues, while a higher level of benzo(a)pyrene phenols was formed by the latter. The relative distribution of benzo(a)pyrene-DNA adducts was similar in all four organs, the major DNA adduct being formed by trans-addition of anti-7,8-dihydroxy-9,10-epoxide-7,8,9,10 tetrahydrobenzo(a)pyrene to the 2-amino group at guanine. These results indicate that the metabolism of benzo(a)pyrene by at least four different organs is qualitatively similar but that quantitative differences exist. PMID- 6277479 TI - Spectrum of tumorigenic phenotypes among adenovirus 2-, adenovirus 12-, and simian virus 40-transformed Syrian hamster cells defined by host cellular immune tumor cell interactions. AB - The tumor-inducing capacities of adenovirus type 2-, adenovirus type 12-, and SV40-transformed LSH hamster embryo cell define a spectrum of four distinct tumorigenic phenotypes: type I, nononcogenic for newborn hamsters but oncogenic for nude mice; type II, oncogenic for newborn hamsters but nononcogenic in syngeneic adults; type III, oncogenic for both newborns and syngeneic adults; and type IV, almost equally oncogenic for syngeneic and allogeneic adult hamsters. Modulation of the cellular immune response of the recipient hamster by immunosuppression or by alloimmunization alters tumor susceptibility, suggesting that dynamic transformed cell-host cellular immune interactions determine the tumorigenic phenotype of a transformed cell line. There is no correlation between the immunogenicities of the transformed cell lines tested and their tumor inducing capacities. However, a strong correlation exists between the ability of transformed hamster embryo cells expressing phenotype IV to produce tumors in allogeneic CB hamsters and their resistance to cytolysis in vitro by activated macrophages. These data suggest that, in addition to transformation, DNA viruses may convey specific tumorigenic phenotypes to the cells that they infect by inducing cellular traits that determine the inherent susceptibility or resistance of a cell to host cell-mediated immune destruction. PMID- 6277480 TI - Natural and antibody-dependent cell-mediate cytotoxicity to cultured target cells superinfected with Epstein-Barr virus. AB - The relationship between natural cell-mediated cytotoxicity (NCMC) and antibody dependent cell-mediated cytotoxicity (ADCC) was examined in an Epstein-Barr virus infected target cell system. The total ADCC reactivity to Epstein-Barr virus infected target cells varied considerably with different effector cells, indicating contributions to specificity by the effector cells as well as by antibodies in the sera. To investigate the role of each reactant, the effector cells, sera, and target cells were tested according to a three-dimensional experimental design and examined for selectivity with the two- and three-way interaction analysis. The two-way analysis was applied to different planes from the experiment to examine special interactions involving two of the three reactants. Selective ADCC was examined through the results from sera versus target cells, selective NCMC was examined by effector cells versus target cells, and the relationship between NCMC and ADCC was examined through the final plane of effector cells versus sera. A three-way interaction analysis applied to the same results supported the conclusions from the two-way analysis and allowed further inquiry into the concurrent role of three reactants. The design and analysis used in the study allowed detection of selective ADCC and NCMC for Epstein-Barr virus-infected target cells and variations in the efficiency of ADCC by different effector cells and in the modulation of NCMC by different sera. PMID- 6277481 TI - Purine and pyrimidine ribonucleoside monophosphate patterns of peripheral blood and bone marrow cells in human acute leukemias. PMID- 6277482 TI - Inhibition of experimental tumor metastasis by selective activation of natural killer cells. AB - The antitumor activity generated by selective activation of natural killer (NK) cells was studied in vitro and in vivo. Unlike Corynebacterium parvum CN6134, which activated both NK cells and macrophages, periodate-oxidized C. parvum CN6134 lost the ability to activate macrophages but retained almost all the NK stimulating capacity of the untreated bacterium. The "inactive" C. parvum strain CN5888 also induced a modest, but selective, activation of NK cells. The enhanced NK cell-mediated cytotoxicity was expressed against YAC-1 lymphoma, UV-2237 fibrosarcoma, and B16 melanoma target cells in vitro and was manifested in vivo by increased destruction of circulating tumor cells and the inhibition of hematogenous tumor metastasis. Periodate-treated C. parvum was as effective in inhibiting the formation of B16 melanoma pulmonary metastases as was untreated C. parvum. In both cases, the inhibiting effect corresponded closely with the kinetics of NK cell activation. PMID- 6277483 TI - Establishment and characterization of an Epstein-Barr virus-negative lymphoma B cell from a patient with a diffuse large cell lymphoma. AB - Cell line (LNPL) was established from a tumor biopsy of a nasopharyngeal "histiocytic" lymphoma composed of large noncleaved B-cells. LNPL expressed immunoglobulin M kappa surface immunoglobulin, as did the original tumor. LNPL also expressed Ia antigen but lacked Epstein-Barr virus nuclear antigen. It exhibited a translocation between chromosomes 8 and 14 which has been described in certain other malignant lymphoproliferative disorders. An autologous T-cell line lacked the translocation between chromosomes 8 and 14. LNPL is being used as an immunogen to produce monoclonal antibodies in our laboratory. Such cell lines may facilitate identification of lymphoma-associated surface antigens. PMID- 6277484 TI - Characterization of a newly established human gastric cancer cell line HGT-1 bearing histamine H2-receptors. AB - A human gastric adenocarcinoma cell line, HGT-1, was established in vitro from the primary tumor of a 60-year-old patient. Histological examination of the tumor revealed a poorly differentiated adenocarcinoma. Primary tumor cells were cloned in soft agarose and gave rise to tumor colonies. The procedures enabling us to form a continuous cell line from the agarose colonies are described. The cultured cells grew as monolayers of closely apposed polygonal cells with a population doubling time of 19.48 +/- 1.20 (S.E.) hr during exponential growth at passage 59. They had an epithelial morphology. Ultrastructural studies revealed the presence of microvilli and tight junctions. The HGT-1 cell line is tumorigenic in nude mice and has a hyperdiploid karyotype with a modal number of 57 chromosomes. It exhibits numerous marker chromosomes. These human gastric epithelial cells do not secrete mucus or carcinoembryonic antigen. They exhibit functional histamine H2-receptors mediating cellular cyclic adenosine 3':5'-monophosphate production and adenylate cyclase activation. In conclusion, the use of a soft-agarose clonogenic assay permitted us to develop a cancer cell line without the problems of fibroblastic cell contamination. The existence of histamine H2-receptors on gastric HGT-1 cells stresses the importance of this line as a model for studies of regulatory mechanisms involved in gastric secretion. PMID- 6277485 TI - Streptozocin: a review of its pharmacology, efficacy, and toxicity. PMID- 6277486 TI - Tumor size, response rate, and prognosis in small cell carcinoma of the bronchus treated by combination chemotherapy. AB - Ninety-eight patients with small cell carcinoma of the bronchus (SCCB) were treated with a combination of cyclophosphamide, methotrexate with calcium leucovorin rescue, and lomustine (CCNU). VP-16-213 was given on relapse. Response rate and survival were analyzed according to extent of tumor, presence of the syndrome of inappropriate antidiuretic hormone production (SIADH), and, in 38 patients, size of intrathoracic tumor as measured by computerized tomographic (CT) scanning. The median survival of the entire group was 34.6 weeks. In patients with limited disease the median survival was 46 weeks compared to 29 weeks for those with extensive disease. This difference in survival between patients with limited and extensive disease was most marked in those showing a partial response (median survival, 45.9 and 28 weeks, respectively). Patients with ectopic antidiuretic hormone production had a poor prognosis even if they had achieved a complete response. Only one partial response to VP-16-213 occurred. There was a strong correlation between initial intrathoracic tumor size as assessed by total cross-sectional area on CT scan and response and survival. When the total area exceeded 30 cm2 there were no complete responders, but there were complete responses in eight of 21 patients when the area was less than 30 cm2. The relationship between intrathoracic tumor size and response rate was present when analyzed according to disease extent, age, sex, and performance status. A clear relationship between total tumor area and survival was also found. When SCCB is treated with chemotherapy alone, the prognosis is better for those patients who present with limited disease, without SIADH, and with small intrathoracic tumor. In this group it is possible that further intensification of therapy may result in prolonged survival. PMID- 6277487 TI - Extensive adenocarcinoma and large cell undifferentiated carcinoma of the lung treated with 5-FU, vindesine, and mitomycin (FEMi): a Southwest Oncology Group Study. AB - Sixty-three previously untreated patients with metastatic non-small cell lung cancer (40 patients with adenocarcinoma and 23 with large cell undifferentiated carcinoma) were treated with combination chemotherapy consisting of 5-FU (300 mg/m2) given as an iv bolus on Days 1-4 and vindesine (3 mg/m2) and mitomycin (10 mg/m2), both given as an iv bolus on Day 1 of each treatment course (FEMi). FEMi was repeated at 3-week intervals for three treatment courses and thereafter at 6 week intervals until disease progression. Major objective responses were seen in 13 of 63 patients (21%). Minor responses were seen in an additional 12 patients (19%). The median survival for all patients was 23 weeks and for responding patients was 38 weeks. The pretreatment performance status had a significant effect on both the response rate and survival time. Patients having an initial Karnofsky performance score greater than or equal to 70% had a 54% response rate, with a median survival of 42 weeks for responding patients. FEMi was well tolerated: 18 patients (29%) did not have any side effects and only five (8%) experienced vomiting. PMID- 6277488 TI - Beta-adrenergic receptors in chronic alcoholic rat hearts. AB - Properties of cardiac beta-adrenergic receptors from chronic alcoholic and control rats were studied to determine whether alterations in the receptor contribute to the decreased responsiveness of isolated working alcoholic rat hearts to beta-adrenergic stimulation. The receptors, assessed in crude membrane fractions by the binding of (-)[3H]dihydroalprenolol, did not differ significantly in either number or affinity in alcoholic rats (22.5 +/- 1.9 fmol . mg protein-1; Kd = 0.49 +/- 0.03 nmol . litre-1; n = 7) compared with control rats (25.9 +/- 1.3 fmol . mg protein-1; Kd = 0.55 +/-0.04 nmol . litre-1; n = 7). Competition experiments indicated that there was no difference in the binding affinity of (-)isoprenaline for the alcoholic and control rat heart receptors, nor in the affinity of (-)propranolol for he alcoholic and control rat heart receptors. In the presence of 5' -guanylylimidodiphosphate, the affinity of ( )isoprenaline for the receptors was decreased the same amount in the alcoholic and control rat hearts. These results suggest that the beta-adrenergic subsensitivity of chronic alcoholic rat hearts is mediated by a biochemical mechanism other than a direct alteration of the beta-adrenergic receptor or coupling between the receptor and adenylate cyclase. PMID- 6277489 TI - [Immunoreactive plasma beta-endorphin in certain physiological situations and pathological conditions (author's transl)]. PMID- 6277490 TI - Morphology and immunocytochemistry of the turtle pituitary gland with special reference to the pars tuberalis. AB - Examination of pituitaries from young and adult turtles representing four families, reveals that in addition to the abundant juxtaneural pars tuberalis (juxPT) found in this class of reptiles, there is generally a substantial amount of pars tuberalis (PT) tissue closely associated with the pars distalis (PD). The PT forms a cortical layer especially conspicuous around the anterior tip of the PD in some species (Trionyx, Kinosternon, Sternotherus), or it forms a thick dorsal layer of tissue irregularly extending onto the sides of the PD in others (Pseudemys, Chrysemys, Lepidochelys, Chelonia). Immunocytochemical studies using unlabelled second antibody and peroxidase-antiperoxidase reveal that in turtles of all ages, the PT tissue allied with the PD (the PTinterna) is composed primarily of cells containing glycoprotein hormones (FSH, LH and TSH), especially the gonadotropins. The juxPT, however, consists mainly of secretory cells unstained by the antisera tested and includes only a small number of gonadotropes and thyrotropes. Although usually widely distributed in the testudinate adenohypophysis, the great majority of gonadotropes and thyrotropes present in the hatchling are in the PTinterna. It is probable that a concentration of these cells in the PTinterna is widespread among vertebrates. In all turtles examined, lactotropes occur principally in the anterior and ventral part of the PD proper; somatotropes are posterior and dorsal. Corticotropes are concentrated as the lactotropes in the anterior PD, but some are also scattered throughout the posterior half of the gland. Lactotropes, corticotropes, and with a few exceptions, somatotropes, do not occur in PT tissue in the turtle. PMID- 6277491 TI - Muscle fibre differentiation and vascularisation in the juvenile European eel (Anguilla anguilla L.). AB - The differentiation of the lateral musculature in the elver stage of the European eel (Anguilla anguilla, L.) has been studied using histochemical staining techniques. Extracellular lipid deposits constitute 12% of the body cross sectional area. Two fibre type may be distinguished on the basis of myofibrillar ATPase activity. Fibres with an alkaline-labile (pH 10.2) ATPase occur as a two fibre layer around the trunk circumference, with invaginations along the horizontal septum and fin insertions. These pale-yellow fibres correspond to the slow ("red") fibres of other fish and comprise around 7% of the body cross section (mean size, 167 micrometer2). Slow fibres show a moderate staining for PAS (glycogen), but a relatively weak reaction of Sudan black (lipid) and the aerobic enzyme markers succinic dehydrogenase (SDH) and cytochrome oxidase (COX). The bulk of the trunk muscle is composed of fast fibres (68%; 328 micrometer2). These are characterised by an alkaline (pH 10.2)-stable ATPase activity, and their innervation. Each fibre is innervated by a single "en-plaque" type endplate at one myoseptal end. Fast fibres adjacent to the slow fibre layer (2-4 fibres deep) show a moderate staining for PAS and a weak reaction for SDH and COX. Deeper regions show a wide range of fibre size, with about 5% being greater than 1,400 micrometer2. Fibres greater than 200 micrometer2 show no significant staining for glycogen, lipid, SDH, or COX. Small fast fibres less than 120 micrometer2 (up to 5% of the white muscle mass) show a strong staining reaction for PAS and a slight reaction for SDH and COX activities. Parameters of vascularisation were calculated from low-magnification electron micrographs (x 1,900). The number of capillaries/fibre and % fibre perimeter in contact with capillaries were, respectively, 0.98, 6.33% (slow fibres); 0.33, 1.96% (superficial fast fibres); and 0.12, 0.71% (deep fast fibres). These values are low in comparison with other fish species. It is suggested that the low aerobic capacity of elver slow muscle reflects a relatively restricted aerobic scope for activity associated with the anguilliform mode of locomotion. PMID- 6277492 TI - Immunohistochemical detection of adenohypophyseal cells containing hormones in Rana ridibunda. AB - The pars distalis of the pituitary of Rana ridibunda captured throughout the spring and summer was examined with immunofluorescence techniques using antisera to mammalian pituitary hormones. On the basis of their immunoreactivity, four different cell types of recognized: 1) cells immunoreactive to anti-bovine LTH, 2) cells immunoreactive to anti-bovine STH, 3) cells immunoreactive to anti-ovine LH, and 4) cells immunoreactive to anti-synthetic ACTH beta (1-24). Their distribution and morphology as well as their staining characteristics (classical histological techniques) are reported in this study. PMID- 6277493 TI - Short interspersed repetitive DNA elements in eucaryotes: transposable DNA elements generated by reverse transcription of RNA pol III transcripts? PMID- 6277494 TI - VSV RNA synthesis: how can you be positive? PMID- 6277495 TI - Infectivity and methylation of retroviral genomes is correlated with expression in the animal. AB - We studied mechanisms controlling gene expression during animal development using retroviruses as model genes. For this, substrains of mice have been previously derived carrying the Moloney leukemia virus (M-MuLV) in their germ line. Virus activation occurs in some of these substrains at different stages of development, resulting in two classes of viral genomes. The genetically transmitted (endogenous) copy is present in every cell, whereas somatically acquired ("exogenous") copies are carried only in cells that were superinfected. We compared these two classes of M-MuLV genomes using two parameters. DNA sequences of the endogenous M-MuLV genome in all mouse substrains were highly methylated in GCGC, the recognition sequence of the restriction enzyme Hha I, and were not infectious (specific infectivity less than 10(-7) pfu per proviral genome) in a DNA transfection assay. In contrast, the "exogenous" copies were hypomethylated and infectious. These parameters are strongly correlated to genome activity in the animal: only tissues carrying exogenous copies express virus-specific RNA. With the assumption that gene expression of transfected DNA is controlled by mechanisms that are relevant for gene expression in the animal, our results suggest that DNA methylation plays a causative role in gene regulation during development and differentiation. PMID- 6277496 TI - Replication of viral DNA sequences integrated within the chromatin of SV40 transformed Chinese hamster lung cells. AB - To determine when during S phase integrated viral DNA sequences in several tsA SV40-transformed Chinese hamster cell clones replicate, we pulse-labeled cultures with BrdUrd and subsequently collected mitotic cells during sequential time intervals. Restriction endonuclease mapping indicated that each of the three SV40 transformed Chinese hamster lung cell clones contained a single viral DNA sequence at a different, but in each case unique, chromosomal site. DNA was extracted from each population of mitotic cells and then was resolved into dense, BrdUrd-containing and light, unsubstituted DNA fractions by cesium chloride gradient centrifugation. In each pair of samples obtained, we measured viral DNA sequences by solution hybridization using single-stranded SV40 32P-labeled DNA probes. Our results support the conclusions that specific genes within a mammalian DNA are programmed to replicate at particular times during S phase, and that the SV40 A gene product, large T antigen, programs integrated viral DNA sequences to replicate very early in S phase. The fractions of viral DNA replicated early in S phase appeared to correlate with each clone's content of functional large T antigen at permissive and nonpermissive culture temperatures. PMID- 6277497 TI - Conservatism in slime mold development. PMID- 6277498 TI - Glucocorticoid regulation of the Ha-MuSV p21 gene conferred by sequences from mouse mammary tumor virus. AB - Molecular chimeras with the p21 transforming gene of Harvey murine sarcoma virus linked to DNA containing the long terminal repeat (LTR) or mouse mammary tumor virus (MMTV) have been constructed. Transformants of NIH 3T3 cells induced by transfection with MMTV LTR-p21 hybrid DNA have been identified that express the normal p21 gene product. The levels of p21 RNA and protein in these transformants are regulated by physiological concentrations of dexamethasone, a synthetic glucocorticoid hormone. Hybrid transcripts containing p21 gene sequences originate at the normal MMTV viral initiation site. It is concluded that sequences necessary for hormonal control of transcription are completely specified by the viral genome and probably map within the viral LTR. PMID- 6277499 TI - Mammary tumor virus DNA contains sequences required for its hormone-regulated transcription. AB - Glucocorticoids regulate the rate of transcription of integrated murine mammary tumor virus (MTV) genes in most clones of MTV-infected rat hepatoma tissue culture (HTC) cells. To determine whether hormonal regulation is mediated from flanking cellular sequences or rather from within the viral DNA, we analyzed the relative rates of transcription of MTV and adjacent HTC sequences in two lines of infected HTC cells, J2.15 and J2.17, each of which contains a single insertion of MTV DNA. In addition, we measured in uninfected HTC cells the transcriptional rates of the corresponding "preinsertion fragments," which may be viewed as the equivalent sequences bearing deletions of MTV DNA; the two genomic segments into which integration occurred in the two lines are unrelated. Previous work showed that glucocorticoids induce the accumulation of MTV RNA in J2.17, whereas viral transcripts are not detected in J2.15, RNA pulse-labeling experiments indicate that glucocorticoids stimulate that rate of MTV gene transcription in J2.17 but not in J2.15; in contrast, no labeled RNA hybridizing to flanking sequences was detected either in the uninfected or in the infected cells. We conclude that the host site of integration cells. We conclude that the host site of integration of MTV need not be transcriptionally active or hormonally responsive to permit viral gene expression. Furthermore, these rate measurements indicate that glucocorticoid-stimulated MTV RNA synthesis in J2.17 does not reflect readthrough transcription from a regulated cellular promoter. This notion is independently supported by transcript mapping experiments showing that the 5' terminus of nuclear MTV RNA is at a site on MTV DNA approximately 1.2 kb downstream from the host-viral junction. Thus our data are consistent with the presence of a hormone responsive element within the provirus. PMID- 6277500 TI - The cloning and reintroduction into animal cells of a functional CAD gene, a dominant amplifiable genetic marker. AB - Rodent cells resistant to PALA, a specific inhibitor of the aspartate transcarbamylase activity of the multifunctional CAD protein, overproduce CAD as a result of amplification of the CAD gene. We cloned a functional CAD gene from Syrian hamster cells using a cosmid vector. Two independently isolated cosmids containing CAD genes have inserts 40 and 45 kb long. We introduced the cloned genes into CAD-deficient Chinese hamster ovary (CHO) cell mutants by fusing them with protoplasts of Escherichia coli containing the cosmids. We also introduced the cloned genes into wild-type CHO cells by selecting cells that became resistant to high concentrations of PALA following protoplast fusion. The transformants of the mutant and wild-type CHO cells contain multiple active copies of the donated Syrian hamster CAD genes. The cloned genes in three independent transformants are integrated into host-cell chromosomes at single locations identified by in situ hybridization. In two of these transformants, the genes are located in one X chromosome or in a chromosome resembling the X. In the third case, the genes are located in a small metacentric or rearranged chromosome. PMID- 6277501 TI - Identification of DNA sequences required for transcription of the human alpha 1 globin gene in a new SV40 host-vector system. AB - We have developed a rapid and simple method for studying the transcription of cloned eucaryotic genes, which involves transfecting SV40-transformed monkey cell lines (COS cells) with derivatives of the plasmid pBR322 that contain the SV40 viral replication origin but lack regions necessary for viral transcription (SV ORI vectors). Because COS cells produce SV40 T antigen and are permissive for SV40 viral replication, transfected SV-ORI plasmids replicate to a high copy number. SV-ORI plasmids carrying a human alpha-globin gene are also replicated in COS cells. Moreover, the alpha-globin gene is faithfully transcribed to produce high levels of RNA, which is accurately processed to produce authentic alpha globin mRNA. We have used this transcription system to demonstrate that a sequence located between 55 and 87 base pairs upstream from the mRNA capping site is required for efficient transcription of the alpha-globin gene in COS cells. PMID- 6277502 TI - Expression of a beta-globin gene is enhanced by remote SV40 DNA sequences. AB - We have studied the transient expression of a cloned rabbit hemoglobin beta 1 gene after its introduction into HeLa cells. Two and one-half days after transfection using the calcium phosphate technique we extracted RNA from the entire cell population and analyzed it by the S1 nuclease hybridization assay. Transcripts were barely detectable when beta-globin gene-plasmid recombinants were used. However, 200 times more beta-globin gene transcripts were found when the beta-globin gene recombinants also contained SV40 DNA, and 90% of these transcripts (about 1000 per cell) had the same 5' end as authentic rabbit globin mRNA. In the latter case, abundant production of beta-globin protein was readily detected in a fraction of transfected cells by immunofluorescent staining. Enhancement of globin gene expression was dependent on SV40 sequences acting in cis, but independent of the viral origin of DNA replication. The enhancing activity was associated with the 72 bp repeated sequence element located at the beginning of the viral late gene region. Viral DNA fragments containing the transcriptional enhancer element could act in either orientation at many positions, including 1400 bp upstream or 3300 bp downstream from the transcription initiation site of the rabbit beta-globin gene. These studies define a class of DNA elements with a mode of action that has not been heretofore described. The activation of genes by specific enhancer elements seems to be a widespread mechanism that may be used for the regulation of gene expression. PMID- 6277503 TI - Structure and evolution of goat gamma-, beta C- and beta A-globin genes: three developmentally regulated genes contain inserted elements. AB - We have determined the complete nucleotide sequence of the goat fetal (gamma), preadult (beta C) and adult (beta A) globin genes. In contrast to other globin genes expressed in different stages of development, these three genes are highly homologous (approximately 90%) in both their coding and noncoding regions. The only major difference between them results from elements inserted into their large introns: gamma contains a 247 bp insertion, beta A contains a 318 bp insertion, and beta C contains both a 252 bp insertion and a 60 bp deletion. Based on comparisons of these three genes to each other and to two goat globin pseudogenes, psi beta X and psi beta Z, we conclude that gamma, beta A and beta C have diverged relatively recently, and that the two gene pairs psi beta X - beta C and psi beta Z-beta A arose via a block duplication of an ancestral pseudogene functional gene pair. Moreover, as opposed to the human fetal genes (G gamma and A gamma), the goat gamma gene appears to be a true beta-like gene that has fetal specific function. The insertions in gamma, beta A and beta C contain sequence features characteristic of transposon-like elements, and appear to be represented in multiple copies throughout the genome. PMID- 6277504 TI - Deletion of the 3' half of the yeast tRNA-Leu3 gene does not abolish promotor function in vitro. AB - The promotor function of the 3' and 5' half tRNA sequences in the yeast tRNA-Leu3 gene has been studied by in vitro transcription in Xenopus laevis germinal vesicle (GV) extracts. Truncation of the DNA template within the tRNA intervening sequence by Hpa I abolishes transcription. However, separation of the tRNA gene halves by insertion of a 300 bp DNA fragment at the Hpa I site does not affect the promoter efficiency. Further, the complete sequence of the 3' half of the tRNA is not necessary for promoter function, because removal of the 3' half of the gene by cleavage with Pvu II, within the DNA inserted at the Hpa I site, does not inhibit transcription. PMID- 6277505 TI - Organization and reorganization of immunoglobulin genes in A-MULV-transformed cells: rearrangement of heavy but not light chain genes. AB - The structure of immunoglobulin-related gene was analyzed in individual Abelson murine leukemia virus (A-MuLV)-transformed lymphoid cell lines. Essentially all of these lines, whether immunoglobulin-containing or null, had DNA rearrangements in the vicinity of the JH regions on both chromosomes as well as deletions of at least 5 kb of DNA 5' to JH. None of these lines, however, except rare light chain producers, had detectable rearrangement at either their kappa or lambda light chain loci. In contrast to A-MuLV-transformants derived from bone marrow. Those from early fetal liver frequently contained more than two and sometimes 12 or more distinct, rearranged JH-containing fragments. Cellular subclones derived from these lines had a subset, usually two, of the fragments found in the parent line. Therefore, heavy chain gene rearrangement appears to precede that of light chain gene rearrangement and is still continuing in certain cultured A-MuLV transformants. PMID- 6277506 TI - The mechanism of cytoplasmic orthopoxvirus DNA replication. AB - Orthopoxvirus DNA replication occurs in the cytoplasm of infected cells within discrete foci designated as virosomes. We show that newly synthesized rabbit poxvirus (RPV) virosomal DNA consists predominantly of concatamers wherein unit length molecules are joined by fusion of two left (LL) or right (RR) ends, resulting in genomes aligned in alternating head-to-head and tail-to tail mirror image arrays. These concatameric molecules serve as the substrates from which unit length DNA molecules are excised during morphogenesis. We propose a mechanism by which internal deletions within these concatameric arrays prior to genome excision and packaging could create inverted terminal repeats and generate gene duplications. PMID- 6277507 TI - Local and spatially coordinated movements in Dictyostelium discoideum amoebae during chemotaxis. AB - We have studied chemotaxis by individual Dictyostelium discoideum amoebae using strong, local gradients of the chemoattractant cyclic AMP. Gradients were provided by diffusion of cyclic AMP from a microneedle, which could be positioned at various points around the cell. Responses to changes in the gradient indicate how the cell is structurally organized for chemotactic movement. There is a polarity in the responsiveness of the surface to stimulation by cyclic AMP along the length of the amoeba. Furthermore, two aspects of chemotactic movement can be distinguished. The first response to cyclic AMP is a locally generated extension of a hyaline pseudopod from the region of the surface nearest the stimulus. The second response, is coordinated and separate from the first response. The coordination appears to depend on the nucleus or on the microtubule-organizing center. PMID- 6277508 TI - Instability of the viral M protein in BHK-21 cells persistently infected with Sendai virus. AB - The study of viral protein expression in BHK cells persistently infected with Sendai virus showed that the viral M protein was greatly reduced in amount or absent in these cells. Pulse-chase experiments demonstrated that the M protein was synthesized at a normal rate, but was unstable compared to the other viral proteins. The M protein instability was independent of temperature and could account for part of the reduction in viral production by persistently infected cells. When a virus stock was grown in embryonated chicken eggs from viruses produced by persistently infected BHK cells, the M protein of this stock presented a restored stability in BHK cells. PMID- 6277509 TI - The plus-strand leader RNA of VSV inhibits DNA-dependent transcription of adenovirus and SV40 genes in a soluble whole-cell extract. AB - In an attempt to determine the mechanism (or mechanisms) by which vesicular stomatitis virus (VSV) kills cells, products of VSV transcription were tested in a cell-free system for their capacity to inhibit transcription of SV40 DNA and plasmids containing adenovirus late promoter and adenovirus-associated RNA genes. VSV RNA transcripts and other RNAs were compared for their capacity to suppress transcription of these DNA templates by RNA polymerases and cofactors present in the HeLa-cell extract system. Relatively low concentrations of the plus-strand leader RNA made in vitro from the 3' end of the wild-type VSV genome were found to inhibit initiation of transcription catalyzed by both RNA polymerase II and RNA polymerase III. Polyadenylated VSV messengers and other natural and synthetic RNAs also caused some inhibitory effects on in vitro transcription from DNA templates, but only at extremely high concentrations. Compared with the wild-type plus-strand RNA leader, the leader RNA synthesized in vitro by defective interfering VSV showed only limited capacity to inhibit RNA synthesis on adenovirus and SV40 DNA templates and only at concentrations at least 30 times greater than that of the wild-type leader. The existence of nucleotide sequences in wild-type leader RNA, not present in defective-interfering leader RNA, that could recognize and block promoters, polymerases or protein cofactors is discussed. PMID- 6277510 TI - Overproduction of the Tn3 transposition protein and its role in DNA transposition. AB - Five single base pair mutations that increase expression of the tnpA (transposase) gene of the Tn3 transposon approximately 30-fold, but which still allow the gene to be regulated, have been isolated by using a generally applicable procedure that involves distally linked lac gene fusions. The mutations, which are all located in a region controlling initiation of translation of the tnpA gene, do not affect normal repression of tnpA by the tnpR gene product, and yield up to a 9000-fold increase in tnpA protein production when combined with a tnpR mutation and placed on a high copy number plasmid. The mutation yielding the highest expression level was separated from the fused lac gene segment by homologous recombination and was found to increase the rate of transposition without altering the nature of the transposition product; in cells defective in both the E. coli recA gene and the tnpR gene of tn3, cointegrate transposition-intermediate structures occur with the overproducing--as well as with the wild-type--tnpA gene. In the presence of a functional Tn3 tnpR gene or the related transposon delta gamma, such cointegrate structures are resolved into the final products of transposition. PMID- 6277511 TI - Analysis of the junction between ribosomal RNA genes and single-copy chromosomal sequences in the yeast Saccharomyces cerevisiae. AB - The yeast Saccharomyces cerevisiae has a single tandem array of 100 ribosomal RNA (rRNA) genes. We have cloned and characterized a junction between the centromere distal end of this array and the adjacent single-copy chromosomal sequences. We have shown that the junction occurs within the nontranscribed region of the repeat. By mapping the junction, we have found that the 35S rRNA precursor is transcribed toward the centromere while the 5S rRNA is transcribed away from the centromere. We have also shown that different yeast strains can have different single-copy sequence at the junction. PMID- 6277512 TI - Selective arrangement of ubiquitinated and D1 protein-containing nucleosomes within the Drosophila genome. AB - We have a new approach, two-dimensional hybridization mapping of nucleosomes, to compare the structures of mononucleosomes from different regions of the Drosophila melanogaster genome. Approximately one in two nucleosomes of the transcribed copia and heat-shock 70 (hsp 70) genes in nonshocked cultured cells contains ubiquitin-H2A (uH2A) semihistone, a covalent conjugate of histone H2A and a small protein, ubiquitin. In striking contrast, less than one in 25 nucleosomes of tandemly repeated, nontranscribed 1.688 satellite DNA contains uH2A, suggesting that most of the nucleosomal uH2A is located in transcribed genes. Approximately 25% of all nucleosomes are ubiquitinated in nonsynchronized cultured Drosophila cells. The hsp 70 genes in nonshocked cells occur in nucleosomes, are greatly enriched in uH2A and are not digested preferentially by staphylococcal nuclease. In contrast, the same genes in chromatin from heat shocked cells are highly sensitive to staphylococcal nuclease and no longer possess nucleosomal organization recognizable with this probe. Histone ubiquitination in transcribed nucleosomes may prevent formation of higher order chromosomal structures by modifying nucleosome-nucleosome interactions. The observed loss of nucleosomal organization in very actively transcribed genes, such as the hsp 70 genes in shocked cells, may be related to the recent finding that ubiquitin conjugates are substrates for the cytoplasmic ATP-dependent proteolytic system. We have also found that 1.688 satellite mononucleotomes contain a specific approximately 50,000 dalton nonhistone protein, D1, in addition to being extremely under-ubiquitinated. D1 may be involved in formation of the highly compact structure of satellite heterochromatin. PMID- 6277513 TI - Adenovirus E1b-58kd tumor antigen and SV40 large tumor antigen are physically associated with the same 54 kd cellular protein in transformed cells. AB - The adenovirus E1b-58kd tumor antigen has been detected in a physical association with a 54 kilodalton cellular protein in adenovirus-transformed mouse cells. Antibody specific for the E1b-58kd protein coimmunoprecipitates a 54 kd protein from transformed, but not from productively infected, cells. Monoclonal antibody specific for the cellular 54 kd protein coimmunoprecipitates the adenovirus E1b 58kd protein from transformed cell extracts. The same or closely related cellular 54 kd protein, associated with the adenovirus E1b-58kd protein, was present in the SV40 large T antigen-54 kd complex previously detected in SV40-transformed mouse cells. The identity of the 54 kd protein is based on the immunological specificities of the anti-54 kd monoclonal antibodies and partial peptide maps of the 54 kd protein associated with the adenovirus and SV40 tumor antigens. The adenovirus E1b-58kd-54 kd complex, like the SV40 large T antigen-54 kd complex, is heterogeneous in size or mass. While all of the cellular 54 kd protein in the adenovirus-transformed cell extract is found in a complex with the E1b-58kd protein, some of the viral 58 kd antigen is detected in a form not associated with the 54 kd protein. The fact that the adenovirus and Sv40 tumor antigens, both required for transformation, can be found in physical association with the same cellular protein in a transformed cell is a good indication that these two diverse viral proteins share some common mechanisms or functions. PMID- 6277514 TI - A membrane-associated precursor to poliovirus VPg identified by immunoprecipitation with antibodies directed against a synthetic heptapeptide. AB - A synthetic heptapeptide corresponding to the C-terminal sequence of the poliovirus genome protein (VPg) has been linked to bovine serum albumin and used to raise antibodies in rabbits. These antibodies precipitate not only VPg but also at least two more virus-specific polypeptides. The smaller polypeptide, denoted P3-9 (12,000 daltons), has been mapped by Edman degradation and by fragmentation with cyanogen bromide and determined to be the N-terminal cleavage product of polypeptide P3-1b, a precursor to the RNa polymerase. P3-9 contains the sequence of the basic protein VPg (22 amino acids) at its C terminus. As predicted by the known RNA sequence of poliovirus, P3-9 also contains a hydrophobic region of 22 amino acids preceding VPg, an observation suggesting that P3-9 may be membrane-associated. This was confirmed by fractionation of infected cells in the presence or absence of detergent. We speculate that P3-9 may be the donor of VPg to RNA chains in the membrane-bound RNa replication complex. PMID- 6277515 TI - The presence of SV40 T-antigen is not required for cytochalasin B-induced multinucleation. PMID- 6277516 TI - Distribution of fibronectin-containing structures on the surface of lamelloplasm and endoplasm of fibroblasts ; hypothesis of receptor-mediated assembly of fibronectin structures. PMID- 6277517 TI - Use of vascular smooth-muscle single cell suspensions for rapid cell number determination in rat thoracic aorta media layer. PMID- 6277518 TI - Reversible inhibition of meiosis resumption in mouse dictyate oocytes by sub optimal media. PMID- 6277519 TI - [The epidemiology, clinical aspects, immunology and morphology of non-A, non-B hepatitis]. PMID- 6277520 TI - [Regulation of blood platelet function by cyclic nucleotides and prostaglandins]. PMID- 6277521 TI - [Histogenesis of storiform tumors (an electron microscopy study)]. AB - The question of histogenetic relations between storiform tumours led to electron optic revision of 29 skin histiocytomas, 10 sclerosing hemangiomas, 13 lenticular dermatofibromas and 10 protuberant dermatofibrosarcomas. Endothelia and pericytes with numerous transient formations, together with various degrees of lipid or some other accumulation on the one hand and fibroproduction tied up with the lateral membrane on the other, were found in all in varying degrees. The filamentous component of tumorous cells had always the properties of myofibrils with a tendency to brushlike formations dependent on the cellular membrane which usually contained numerous pinocytotic vacuoles. It thus differed from the cells of storiform neurofibromas (4 cases), whose intracytoplasmatic filaments showed no tendency to clustering or relation to the cellular membrane. Moreover, in their finer lateral membrane no substantial production of collagenous fibers occurred. According to the results of the study, histiocytoma, sclerosing hemangioma and dermatofibroma belong to the same category of tumours from mixed vasoforming tissue. Of the same, but mostly hemangiopericytary, origin is probably also protuberant dermatofibrosarcoma provided it is histogenetically uniform. Storiform neurofibroma is structurally different. PMID- 6277522 TI - Potentiation of cell killing by inhibitors of poly(ADP-ribose) polymerase in four rodent cell lines exposed to N-methyl-N-nitrosourea or UV light. AB - The sensitivities (Do-values) of the cytotoxic effect of MNU on four rodent cell lines were: mouse L1210, 0.07 mM; rat Yoshida sarcoma, 0.52 mM; Chinese hamster V79A, 0.70 mM and the UV sensitive, X-ray sensitive V79/79, 0.35 mM. The abilities of maximum non-toxic doses of the poly-(ADP-ribose) polymerase inhibitors, 5-methyl nicotinamide (5MeN), 3-methoxybenzamide (3MBA) and caffeine to potentiate this cytotoxicity and that of UV light in V79A and V79/79 was measured. The degree of potentiation (ratio Do without inhibitor/Do with inhibitor) was both agent and cell line dependent. In general the lymphoid cell lines L1210 and YS showed greater potentiation, up to 4-fold, than did the fibroblast lines V79A and V79/79. The use of inhibitors in pairs suggested that 5MeN and 3MBA affect one process whereas caffeine affects additional processes. The data provide further support for a role for poly(ADP-ribose) in DNA repair, but indicate that metabolic factors may modify the effectiveness of individual inhibitors of poly(ADP-ribose) polymerase in different cell lines. PMID- 6277523 TI - Associations of benzo[a]pyrene with rat-liver chromatin and the chromatin protein. AB - Rat-liver nuclei were prepared in the course of time after the i.p. injection of [G-3H]benzo[a]pyrene ([3H]BP). The nuclei were lysed in the hypotonic buffer and centrifuged at 4000 X g. The recovery of the radioactivity of resulting supernatant (chromatin) was thus 91% at 24 h, 68% at 48 h and 74% at 168 h after the i.p. injection. The incorporation into nucleosome-oligomer fraction was always much more than into those of monomer and DNA-rich fractions. The preferential incorporation was found in the fraction which was enriched in non histone chromatin proteins (NHCPs) of 49 000-55 000 daltons. This fraction steadily raised the incorporation level until at 168 h after the i.p. injection. In contrast, the levels of histone and DNA fractions were always very low. The significant incorporation was observed in the fraction which was composed of five classes of histones and low molecular-weight NHCPs (less than 30 000 daltons), despite the very low incorporation into the histone fraction. The fluorographic analysis revealed the predominant incorporations at the positions of molecular weight of 65 000, 52 000 and 44 000 daltons. In addition, the incorporations were clearly observed at the positions of 59 000, 49 000, 45 000, H1 histone, A24 protein and another one. On the other hand, these fractions were, at the final preparation steps, subjected to either dialysis of SDS-phenol treatment and/or acetone precipitation. The total recovery of radioactivity was thus 21% at 24 h, 32% at 48 h and 52% at 168 h after the i.p. injection. These results suggest that the chromatin contains considerable amounts of water-soluble, phenol and/or acetone-soluble BP-conjugates in the early period after the i.p. injection. PMID- 6277524 TI - The effect of benzo[a]pyrene on sponges as model organisms in marine pollution. AB - The majority of the investigations were performed with the marine sponge Tethya lyncurium at concentrations of 2 X 10(-8) to 1 X 10(-11) g/ml of benzo[a]pyrene (BaP). Sea-pollution was characterized as BaP equivalent activity in the Ames test. Increased activity of ornithine decarboxylase (ODC) was observed when sponges were artificially exposed at polluted marine areas for 3 weeks. In contrast to the situation in higher animals no ODC induction of the fast type was observed. Mixed function oxygenases (MFO) were not detected in sponges nor could they be induced as in vertebrates. BaP was absorbed by Tethya and concentrated 30 -60-fold. In live, but not dead, artificially perfused sponges [3H]- and [14C]BaP radiolabeled became firmly associated with DNA, RNA and protein of the sponges. The association persisted in isolated fractions, in nucleotides, in nucleosides and in protein hydrolysates. The BaP binding ratio to DNA was found to be strongly correlated to the concentration of BaP. Light modifies BaP and thus enables binding. In the dark only very low association, if any, is observed. The possible consequences of these findings are discussed. PMID- 6277525 TI - Conformational study of purified epoxide hydrolase from rat liver. AB - Hepatic epoxide hydrolase (EC 3.3.2.3) was purified from phenobarbital-treated rats by ion-exchange chromatography followed by hydrophobic chromatography. The enzyme had a specific activity of 300--400 nmol min-1 mg-1 protein with benzo[a]pyrene-4,5-oxide as the substrate. Circular dichroism (CD) spectra of the purified enzyme gave two negative bands, centered at 210 nm and 222 nm, respectively. The mean residue ellipticity at 222 nm was 12,9000 deg X cm(2) X dmol(-1), which indicated the presence of about 35% alpha-helical structures. Sodium dodecyl sulfate (SDS) greatly affected the shape of the CD spectra, which were gradually shifted to the blue. This suggested a decrease in the aggregation state of the protein. Electrostatic interactions were important in the organization of the enzyme structure since the conformation was stable between pH 7.4 and pH 10. At pH-values 5.0, 6.0 and 12.0, the CD bands underwent considerable changes in both amplitude and shape. Moreover there was a good correlation between the optimal pH range of the epoxide hydrolase activity and the organization state of the protein. After membrane reconstitution with liposomes, the conformation of the enzyme was not significantly modified by the presence of dimyristoyl L-alpha-phosphatidylcholine or other phospholipids. This constancy was obtained over a wide range of molar ratios of phospholipids to protein (0--500). However, phospholipids did increase the thermal stability of the enzyme. Fluorescence measurements of diphenylhexatriene (DPH) bound to dimyristoyl L-alpha-phosphatidylcholine indicated that addition of epoxide hydrolase modified the thermal transition of the lipid phase. On the other hand, electron paramagnetic resonance (EPR) signals of the nitroxide-labelled fatty acid, 2-(14-carboxy-tetradecyl)-2-ethyl-4,4-dimethyl-3,3,-oxazolidiny-oxyl, bound to the phospholipid, indicated that the presence of the protein decreased by about 53% the correlation time of the label, suggesting that its motion had increased. In conclusion, phospholipid-epoxide hydrolase interactions enhanced the fluidity of dimyristoyl L-alpha-phosphatidylcholine liposomes without changing the secondary structure of the enzyme. Electrostatic interactions also played an important role in the conformational stability of the protein. PMID- 6277526 TI - Mechanisms of quinidine-induced depression of maximum upstroke velocity in ovine cardiac Purkinje fibers. AB - A major advance in understanding how quinidine depresses maximum upstroke velocity (Vmax) is the Hondeghem-Katzung mathematical model which incorporates voltage-independent rate constants for binding to and unbinding from resting, open, and inactive Na channels, and a voltage shift of -40 mV for the Hodgkin Huxley h-kinetics of quinidine-associated Na channels. Using a double microelectrode voltage clamp technique to control transmembrane voltage and apply conditioning pulses, we found that quinidine blockade increased as transmembrane voltage became more positive in the range -60 to +40 mV, and that the rate of quinidine dissociation increased as transmembrane voltage became more negative in the range -60 to -140 mV. The relationship of Vmax to transmembrane voltage obtained at drive cycles from 500 msec to 20 seconds conformed to the model modified to include voltage-dependent rate constants without the postulated -40 mV shift for quinidine-associated channels. Thus binding of quinidine to inactive Na channels and unbinding from resting channels are both voltage-dependent and can explain frequency and voltage dependent actions of quinidine on Vmax without any voltage shift for quinidine-associated channels. PMID- 6277527 TI - Vascular smooth muscle. Calmodulin and cyclic AMP-dependent protein kinase after calcium sensitivity in porcine carotid skinned fibers. AB - Recent work on vascular smooth muscle actomyosin has indicated that the Ca2+ sensitivity of both ATPase and superprecipitation are affected by calmodulin (CaM) and cyclic AMP-dependent protein kinase (cPK). Using a "chemically skinned" arterial preparation, we have extended these observations to the intact structured contractile system. Media from hog carotid artery were skinned with 1% Triton X-100 followed by a 50% glycerol-ATP salt solution, in which the strips were stored at -25 degrees C. Small strips (thickness between 0.1 and 0.2 mm) were mounted isometrically and relaxed in a Mg-ATP salt solution, pH 6.7, Ca2+ 10(-8) M, 30 degrees C. Ca2+ elicited a contraction with an ED50 of 10(-6) M. Isometric force was between 1 and 4 mN, consistent with the force observed before skinning. With time, the preparation became less sensitive with an increase in ED50 to 10(-5.7) M. CaM (4 micro M) reverses this loss, stabilizes the preparation, and sharply accelerates the rate of tension development. The ED50 in the presence of 4 micro M CaM shifts to about 10(-7) M. This effect is dose dependent, with the half maximal effect at about 0.4 micro M CaM. Submaximal Ca2+ contractions can be reversibly depressed by preincubation of relaxed fibers with cPK catalytic subunit (300 U/ml), even in the presence of 4 micro M CaM. An inhibition of about 50% of the contraction at 0.2 micro M Ca2+ was obtained, whereas only 20% inhibition was found at 6 micro M Ca2+. Our findings suggest that changes in vascular contractility cannot be described solely in terms of changes in cytoplasmic Ca2+, and that changes in the sensitivity of the contractile protein to a given Ca2+ concentration are also potential mechanisms for vasodilation. PMID- 6277528 TI - Contrast enhancement of myocardial infarction: dependence on necrosis and residual blood flow and the relationship to distribution of scintigraphic imaging agents. PMID- 6277529 TI - Recent advances in the control and function of the anterior pituitary. AB - Major advances in our understanding of the synthesis and release of anterior pituitary hormones have been made over the past several years. Neurons of the hypothalamus have been found to serve as "neuroendocrine transducers" in that they have both electrical and secretory functions. Peptidergic neurons respond to appropriate stimuli with a release of hypothalamic factors into the hypophyseal portal system. These factors or hormones ultimately control the endocrine function of anterior pituitary cells. Three hormones, Thyrotropin Releasing Hormone (TRH), Gonadotropin Releasing Hormone (GnRH or LHRH) and somatostatin have been identified, synthesized and tested for clinical applications. The clinical assessment of pituitary function has been greatly improved by new and improved radioimmunoassays. One of the recent clinical advances in the area of pituitary disease has been the determination of the relatively high frequency of prolactinomas. Prolactin secreting microadenomas are an important and treatable cause of amenorrhea and infertility in young women. In addition, many pituitary tumors previously believed to be non-functional or "chromophobe adenomas" appear to be prolactinomas. Many new diagnostic and therapeutic techniques are continuing to be developed to improve our management of patients with hypothalamic-pituitary disease. PMID- 6277530 TI - Interactions among endocrine control systems in the regulation of ovarian function. AB - During sequential physiologic maturation of an individual follicle, the number of granulosa cells increases in excess of 1000-fold, while intra-ovarian concentrations of sex steroids escalate by 100-fold. The recent development of several in vitro ovarian systems has permitted a more extensive and direct assessment of specific mechanisms that control growth and steroidogenesis in granulosa and thecal cells. For example, estradiol and follicle stimulating hormone seem to promote the production by ovarian cells of low-molecular-weight growth factors, that may participate in granulosa cell proliferation. Luteinizing hormone stimulates the de novo synthesis of prostacyclin by granulosa cells in vitro. Prostacyclin, in turn, may regulate the microvasculature of the maturing follicle and directly stimulate steroidogenesis. The effects of individual hormones are markedly modified by other intraovarian endocrine factors, and by the precise status of cytodifferentiation of the ovarian cells. For example, the actions of prolactin on granulosa-cell steroidogenesis are influenced strikingly by both agonistic and antagonistic interactions between prolactin and estradiol, as well as by the level of granulosa-cell cytodifferentiation attained in vivo. Similar bihormonal intrafollicular interactions are recognizable between estrogen and follicle-stimulating hormone in the early follicle, and between estradiol and luteinizing hormone in the maturing follicle. These interactions are susceptible to more precise examination under defined in vitro conditions. Overall, recent advances in biomedical research continue to elucidate basic molecular mechanisms of hormone action in ovarian cell physiology. Such advances are likely to continue to provide important insights into the pathophysiology of clinical disorders of human reproduction. PMID- 6277531 TI - Studies on pyrophosphate diesterase activity in cultured human fibroblasts: a deficiency in Niemann-Pick disease. AB - Fibroblast phosphodiesterase activity was studied using 4-methylumbelliferyl pyrophosphate diester as substrate. Release of the fluorogen, 4 methylumbelliferone, was found to be dependent on acid phosphatase activity, normally present in excess in crude cell extracts. Phosphodiesterase activity had an acid pH optimum, was deficient in Niemann-Pick disease fibroblasts, and, when assayed in the presence of exogenous acid phosphatase, had an identical electrofocusing profile to that of sphingomyelinase. These findings suggest that 4-methylumbelliferyl pyrophosphate diesterase and acid sphingomyelinase activities are dependent on the same enzyme. PMID- 6277532 TI - A new radiochemical assay for fructose-1,6-diphosphatase in human leucocytes. PMID- 6277533 TI - Determination of estrone sulfate in plasma by radioimmunoassay without deconjugation. AB - A radioimmunoassay method using highly specific antiserum without prior deconjugation has been developed for determination of estrone sulfate in human plasma. Antiserum was elicited in the rabbit by immunization with antigen in which the sulfated steroid hapten is linked to a carrier protein through the C-6 position. After treatment with Rivanol, the protein of albumin-free antiserum was covalently bound to a p-arylamine glass bead support through the cross-linkage with glutaraldehyde. A simple and reliable method for measurement of estrone sulfate which involves elimination of endogenous interferences by use of Sep-pak C18 followed by radioimmunoassay employing the antibody-glass preparation, was established and applied to peripheral plasma throughout normal pregnancy. PMID- 6277534 TI - Hepatic Disorders. PMID- 6277535 TI - Disorders of mineral metabolism. PMID- 6277537 TI - Lp(a) lipoprotein enters cultured fibroblasts independently of the plasma membrane low density lipoprotein receptor. AB - Lp(a) lipoprotein shares the apoB antigen with low density lipoprotein (LDL). The Lp(a) antigen is unique for Lp(a) lipoprotein. Fibroblast association (i.e. plasma membrane binding plus intracellular accumulation), plasma membrane binding, intracellular accumulation and degradation of 125I-Lp(a) lipoprotein were studied in strains from subjects with or without autosomal dominant hypercholesterolemia (HC). Subjects without HC (non-HCs) have cell surface receptors for low density lipoprotein (LDL receptors). On the average, HC heterozygotes have half-normal LDL receptor activity and "receptor-negative" HC homozygous cell strains lack functional receptors. Fibroblast processing of 125I Lp(a) lipoprotein was compared to fibroblast processing of 125I-LDL. LDL receptor dependent processing of 125I-LDL was saturated at about 50 microgram apo 125I LDL.ml-1 in non-HC fibroblasts. 125I-Lp(a) lipoprotein was, however, largely processed independently of receptor mechanisms by non-HC cells (highest concentration examined 150 microgram apo 125I-Lp(a) lipoprotein . ml-1). Lp(a) lipoprotein did not interfere with 125I-LDL for fibroblast association, but inhibited 125I-LDL degradation. The interference with 125I-LDL degradation was time dependent. Only slightly higher 125I-Lp(a) lipoprotein processing values were found in non-HC and HC heterozygous strains than in "receptor-negative" HC homozygous strains. However, non-HC cells had more than tenfold higher 125I-LDL processing values than "receptor-negative" HC homozygous cells. PMID- 6277536 TI - Low density lipoprotein receptor activity in cultured fibroblasts from subjects with or without ischemic heart disease (in the absence of familial hypercholesterolemia). AB - Fibroblast strains from six subjects with ischemic heart disease (IHDs) were compared to strains from 43 subjects without a history of IHD (non-IHDs), with respect to association (plasma membrane binding plus intracellular accumulation) and degradation of radioiodinated LDL (125I-LDL). The subjects (25 females and 24 males) were selected on the criteria that they were twins (one from each pair), 58-61 years old, and living within 200 km of Oslo. None of them suffered from autosomal, dominant hypercholesterolemia, which is associated with reduced cell surface LDL receptor activity and increased susceptibility to IHD. There was a trend towards lower 125I-LDL association values in strains from IHDs than in strains from non-IHDs (P=0.009). There was a significant negative correlation between, on one hand, serum total cholesterol level and on the other fibroblast association (P=0.03) or degradation (P=0.04) of 125I-LDL. We have previously presented data indicating that fibroblast association of LDL may be determined by alternate genes at one single locus. Together with the present limited data, this raises the possibility that normal genes at the LDL receptor locus may render subjects more or less susceptible to ischemic heart disease. PMID- 6277538 TI - Inherited renal disease and genetic counseling. AB - Inherited renal abnormalities and diseases are less common than acquired disorders. However, they are of great interest because their study results in increased understanding of the embryogenesis and physiology of the kidney, the pathogenesis of acquired disease, improved therapeutic approaches and accuracy of genetic counseling. Inherited defects of the kidney may be structural, functional or part of genetically transmitted systemic diseases that have major effects on renal structure and/or functions. Most structural defects of the kidney, with the exception of varying forms of cystic disease and the hereditary nephritides, are congenital and only rarely inherited in a Mendelian sense. The majority of genetically transmitted abnormalities of proximal and distal tubular function are caused by inborn metabolic errors or enzyme defects and deficiencies. Amniocentesis, ultrasonography and enzymatic assays have made the prenatal diagnosis of many inherited renal diseases possible so that more accurate counseling early therapeutic intervention may be provided. PMID- 6277539 TI - The effect of long-term chlorpromazine and desipramine treatment on adrenal corticosterone, cyclic AMP, and cholesterol distribution in rats. AB - 1. Male rats were injected intraperitoneally with chlorpromazine (10 mg/kg) and desipramine (10 mg/kg), respectively, twice daily for 2 weeks. 2. The adrenal corticosterone concentrations were significantly lowered in chlorpromazine- and desipramine-treated rats. 3. Large dose of corticotrophin would increase the adrenal corticosterone level in chlorpromazine-treated but the adrenal response was significantly less than in the untreated controls. 4. The adrenal cyclic AMP level was similar in treated and control rats and the adrenal response to corticotrophin was not decreased at the cyclic AMP level. 5. The cholesteryl ester concentrations in the adrenals were significantly increased by drug treatment, although the free cholesterol concentrations were unchanged in the cytosol and mitochondrial cell fractions. 6. The possible site of action of chlorpromazine in suppressing corticosterone production is discussed. PMID- 6277540 TI - [Dipterex polyneuropathy--ultrastructural study of the axonal change (author's transl)]. PMID- 6277541 TI - Accumulation of MDP in hepatic metastases from mucinous adenocarcinoma of the colon. AB - The case of a 66-year-old man with mucinous adenocarcinoma of the rectum with metastases to the liver is presented. Several mechanisms of accumulation of MDP are identified and correlated with CT scan findings. PMID- 6277542 TI - Combined nuclear medicine and ultrasound studies in the evaluation of suppressed thyroid tissue. PMID- 6277543 TI - Incidence and consequences of breast artifacts in radionuclide cardiac studies. AB - A retrospective evaluation was done on 426 radionuclide cardiac studies performed on female patients at our institution from January through September, 1980. These procedures included pyrophosphate (PYP) myocardial scans, resting and exercise Tl 201 studies, and gated equilibrium studies. The purpose of this evaluation was to document the incidence and possible consequences of breast artifacts in various cardiac procedures. We found the incidence to range from 30 to 50% of the studies performed, and both the incidence and type of artifact encountered varied with the type of procedure performed. Furthermore, 13 to 35% of patients had artifacts directly overlying the myocardium. The possible consequences, including potential misdiagnosis, interference with computer derived quantitative parameters, and the technical maneuvers necessary to avoid or alleviate these problems are discussed. PMID- 6277544 TI - A case of traumatic arteriovenous fistula diagnosed by radionuclide cerebral scan. PMID- 6277545 TI - Converting enzyme inhibition with captopril in patients with primary hyperaldosteronism. AB - The humoral and hemodynamic effects of converting enzyme inhibition captopril are presented in two patients with primary hyperaldosteronism (PHA). In all, 20 patients with resistant hypertension were treated with the angiotensin converting enzyme inhibitor captopril. In 18 patients with essential or renovascular hypertension mean (+/- SEM) plasma renin activity (PRA) rose from 5.0 +/- 1.4 to 35.3 +/- 5.3 ng/ml/hr (P less than 0.01) and mean (+/- SEM) plasma aldosterone (PA) declined from 25.8 +/- 2.9 to 15.1 +/- 1.9 ng/ml (P less than 0.01) after captopril. In two patients with PHA the PRA was not stimulated by converting enzyme inhibition, although there was modest decline in PA and a temporary reduction in blood pressure. After surgical removal of aldosterone-producing adenomas, PRA responsed appropriately to captopril. These cases illustrate that a disease process can modify the response to a drug and demonstrate that, in patients with PHA, captopril does not stimulate PRA, induces only minor decrements in PA, and is relatively ineffective as an antihypertensive. PMID- 6277546 TI - Ceftizoxime kinetics and renal handling. AB - The kinetics and renal handling of ceftizoxime were examined after intravenous and intramuscular injection and the effect of probenecid on its excretion was investigated. Peak serum level after 1000 mg IV was 60.5 microgram/ml and half life (t 1/2) was 1.4 hr. Peak serum level (40.9 microgram/ml) was reached 1 hr after 1000 mg IM. When probenecid was added to the 1000-mg IM dose the peak level was 44.3 microgram/ml at 1 hr and serum levels at 2, 4, 6, and 8 hr were all higher than after ceftizoxime alone (P less than 0.01). The 1.85-hr t 1/2 of ceftizoxime alone was extended to 2.29 hr when probenecid was added. Ceftizoxime was shown to be actively secreted by the renal tubule; this secretion was decreased by probenecid. PMID- 6277547 TI - Cholesterol transport through the plasma. PMID- 6277549 TI - Neurotoxic metabolites of "commercial hexane" in the urine of shoe factory workers. AB - Urinary metabolites were tested in 41 shoe-factory workers exposed to a mixture of 10 solvents among which "commercial hexane" was the prevailing component. Cyclohexanol, 2-methyl-2-pentanol, 3-methyl-2-pentanol, and trichloroethanol were determined in connection with exposure to cyclohexane, 2-methylpentane, 3 methylpentane, and trichloroethylene, respectively. 2-Hexanol, 2,5-hexanedione, 2,5-dimethylfuran, and gamma-valerolactone were all determined in connection with n-hexane exposure only. 2,5-Hexanedione was the principal n-hexane metabolite found in the workers' urine. This finding of the experimentally proven neurotoxin 2,5-hexanedione in the urine of shoe-factory workers exposed to "commercial hexane" is consistent with the idea that this compound is responsible for the development of neuropathy in this group of individuals. PMID- 6277548 TI - Experimental neurotoxicity and urinary metabolites of the C5-C7 aliphatic hydrocarbons used as glue solvents in shoe manufacture. AB - Rats were intermittently exposed (9 to 10 h/d, 5 to 6 d/week) to controlled concentrations of single analytical grad solvents in ambient air. After periods ranging from 7 to 30 weeks the animals were perfused with glutaraldehyde and samples of nerves were processed for light microscopy of sections and of teased fibers. Animals treated with n-hexane at 5000 ppm (14 weeks) or 2500 ppm (30 weeks) developed the typical giant axonal degeneration already described in rats treated continuously with 400 to 600 ppm of the same solvent for 7 weeks or more. No such alterations were found in rats subjected to the following intermittent respiratory treatments: n-hexane 500 ppm (30 weeks) or 1500 ppm (14 weeks), cyclohexane 1500 or 2500 (30 weeks), n-pentane 3000 ppm (30 weeks), n-heptane 1500 ppm (30 weeks), 2-methylpentane 1500 ppm (14 weeks), and 3-methylpentane 1500 ppm (14 weeks). The following metabolites were found in the urine of rats according to treatment (in parenthesis): 2-methyl-2-pentanol (2-methylpentane); 3 methyl-2-pentanol and 3-methyl-3-pentanol (3-methylpentane), 2-hexanol, 3 hexanol, gamma-valerolactone, 2,5-dimethylfuran, and 2,5-hexanedione (n-hexane). 2-Hexanol was found to be the main urinary metabolite of n-hexane, while 2,5 hexanedione was present only in a lesser proportion. This feature of rat metabolism suggests that in this species 2,5-hexanedione reaches an effective level at its site of action during intermittent respiratory treatment with n hexane with difficulty and explains the high concentrations necessary to cause polyneuropathy in rats subjected to this treatment. PMID- 6277550 TI - A comparative study of the toxicity of n-pentane, n-hexane, and n-heptane to the peripheral nerve of the rat. PMID- 6277551 TI - Perhexiline maleate induced lipidosis in human peripheral nerve and tissue culture: ultrastructural and biochemical changes. PMID- 6277552 TI - A biochemical assessment of the neurotoxicity of the radiosensitizing drug misonidazole in the rat. AB - We have obtained biochemical evidence that misonidazole when administered in large doses to rats produces a sparse dying-back peripheral neuropathy and degenerative changes in the trigeminal ganglia and cerebellum. In our experience these neurotoxic effects of misonidazole cannot be detected reliably by the use of simple behavioural and functional tests, e.g., inclined plane and narrowing bridge tests (Rose and Dewar, unpublished results). Therefore, these methods would be of limited use in the neurotoxicity screening of misonidazole analogues. On the other hand, the biochemical approach provides a convenient quantitative method which could be used as the basis for comparing the neurotoxicity of other candidate radiosensitizing drugs. PMID- 6277553 TI - Late peripheral neuropathy due to an acute voluntary intoxication by organophosphorus compounds. PMID- 6277554 TI - Effects of ganglioside therapy on experimental CS2 neuropathy. AB - Both in animals and in man the inhalation of CS2 vapor induces a chronic polyneuropathy with primary lesions in the axons of peripheral nerves. Since it was reported in several studies that the administration of gangliosides improves nerve regeneration and the functional recovery of nerves damaged by section as well as cryodegeneration, a study was undertaken to evaluate the effects of bovine-brain gangliosides administration on the experimental CS2 neuropathy in the rat. One hundred and fifty male rats were intoxicated with CS2 by a discontinuous inhalation exposure to 700 ppm for 12 weeks until a clear neuropathy developed. Thereafter the animals were subdivided at random into five groups and treated in different ways: 10 mg/kg BW gangliosides, 0.5 mg/kg gangliosides, 0.5 mg/kg vitamin B1, and 1 mg/kg vitamin B6, physiological solution, and controls without any treatment. The recovery from neuropathy was controlled for 18 weeks of treatment and assessed periodically by means of clinical, electromyographic, and morphological examination. The results of morphological studies showed more pronounced regeneration activity in the rats treated with the high dose of gangliosides than in all others, while no differences among the groups could be observed as far as clinical and neurophysiological parameters are concerned. The mechanism supporting this ganglioside-induced effect has so far not been ascertained, and further studies on this subject are in progress. PMID- 6277555 TI - Cyanide and its relationship to nervous suffering. Physiopathological aspects of intoxication. AB - On the basis of personal results in man and laboratory animal, we examined some aspects of acute and chronic cyanide intoxication: physiopathological aspects, hydroxocobalamin's therapeutic action in treating acute intoxications and neurotoxic sufferings due to cyanide, and hydroxocobalamin's biological action in decreasing the level of free cyanide appearing in blood and urine, a few days after sciatic nerve clamping in rabbits. PMID- 6277556 TI - Viral diarrhea of young animals: a review. PMID- 6277557 TI - [Evolution of anti-rota virus antibodies in the milk of cows treated in the last month of pregnancy either by adjuvated rotavirus vaccine or by the adjuvant fraction of the vaccine (author's transl)]. PMID- 6277558 TI - Susceptibility of cell line cultures of bovine kidney origin to infectious bovine rhinotracheitis and parainfluenza-3 viruses. PMID- 6277559 TI - Cerebral fluoride in mania and the effect of lithium: a hypothesis. PMID- 6277560 TI - Effects of acute and chronic administration of phencyclidine on dopaminergic receptors in rat striatum. PMID- 6277561 TI - NAD glycohydrolase: enzyme characterization using intact mammalian erythrocytes. AB - 1. NADase activity has been determined on intact erythrocytes of several species. 2. Although a wide range in maximum velocity exists across species, Michaelis constants observed are very similar. 3. The enzyme is found on the outer surface of the erythrocyte plasma membrane. 4. It is inhibited by substrate after an apparent permanent modification. 5. This modification may be due to self ADP ribosylation. 6. We have also demonstrated the presence of an ADP ribosyltransferase on the outer surface of the sheep erythrocyte membrane. PMID- 6277562 TI - Phylogeny and ontogeny of the phosphoglycerate mutases - III. Inactivation of rabbit muscle phosphoglycerate mutase (type M isozyme) by the sulfhydryl group reagents. AB - 1. The three phosphoglycerate mutase isozymes from mammals (types M, B and MB isozymes) differ in their sensitivity to the - SH group reagents. 2. Rabbit muscle phosphoglycerate mutase (type M isozyme) is reversibly inactivated by tetrathionate, rho-chloromercuribenzoate and Hg2+. 3. Titration with rho chloromercuribenzoate shows the existence of two sulfhydryl groups per enzyme subunit, the modification of which produces a progressive decline in enzyme activity. 4. The apparent Km values for substrate and cofactor are not affected by tetrathionate treatment. 5. Phosphoglycerate mutase inactivated by tetrathionate and by rho-chloromercuribenzoate is unable to form the functionally active phosphorylenzyme when mixed with glycerate-2,3-P2, and is not protected by the cofactor against heating. 6. Glycerate-2,3-P2 protects against tetrathionate treatment, but fails to protect against Hg2+ and rho-chloromercuribenzoate inactivation. PMID- 6277564 TI - Radionuclides in the diagnostics of ischaemic heart disease. Introduction. PMID- 6277563 TI - Guaiacol peroxidase levels in human cervical mucus: a possible predictor of ovulation. PMID- 6277565 TI - Changes in total and local contractility of the left ventricle, imaged by radionuclide ventriculography, in patients with ischaemic heart disease during submaximal exercise. AB - In 40 patients with chronic ischaemic heart disease [IHD) and 10 healthy subjects the total and local myocardial contractility was assessed during threshold-level exercise. The patients were subjected to radionuclide ventriculography in a modification recording the first passage of the radioindicator through the heart cavities. In the control subjects the exercise induced a 27% increase in the total ejection fraction, with increased contractility of all left ventricular segments. In the patients with IHD the reactions to exercise varied. In 40% of the patients the total ejection fraction increased adequately, in 17.5% the increase was nonadequate, in 20% no increase was observed, and in 22.5% the ejection fraction decreased. The most marked reduction in myocardial contractility was found in patients who had sustained myocardial infarction and reacted positively to the exercise test. In 34 patients there were observed left ventricular segments showing a paradoxical response of contractility to exercise- a decrease in the local ejection fraction. This pathological local reaction occurred rather in a normal function at rest than in a myocardium with hypokinetic segments at rest. PMID- 6277566 TI - Myocardial scintigraphy with 99m-Tc-pyrophosphate in patients with acute myocardial infarction. AB - The purpose of the study was to find the optimal conditions for scintigraphic examination in acute myocardial infarction (AMI), as well as to develop an algorithm and a computer programme for a more objective processing of the results obtained with gamma camera, and, finally, to juxtapose the evidence obtained and the clinical pattern. Thirty-six patients aged 34-80 years (28 with AMI and 8 with chronic ischaemic heart disease) were examined with the aid of a gamma camera. The scintigrams were evaluated visually by three experts independently and were assigned scores from 0 through 3. They were also processed with a computer. Maximal indicator accumulation was found by the end of hour 2 after injection. The indicator accumulation was higher on days 3-5 after the onset of the condition as compared with that on the first two days. The accumulation extent depends on the severity and, in some projections, on the location of the AMI as well. The scintigraphic imaging and assessment of the severity of AMI is reliable and useful, especially when the findings are processed with a computer. PMID- 6277567 TI - 99mTc-labelled pyrophosphate myocardial scanning in experiment and in clinical practice. AB - A survey is presented of research results of pyrophosphate myocardial scanning in 130 dogs and 150 patients with various myocardial lesions. The results of experiments in animals were immediately applied in clinical practice. The results showed that acute transmural myocardial lesions yielded practically in all instances positive scintigraphic findings. The findings in nontransmural infarctions were less convincing--they were positive in 60% of cases. Positive scintigraphic findings were also obtained in 80% of dogs with experimentally induced temporary ischaemia, and in 80% of dogs with experimentally induced dystrophy of the myocardium. In patients with angina pectoris positive findings were obtained in 1/3 of the cases, and in patients with cardiomyopathies, in 1/3 to 1/4 of the cases (in dependence on the etiology of the cardiomyopathy). It appears that 99mTc-PYP is a highly sensitive, but nonspecific, detector of myocardial lesions. PMID- 6277568 TI - Characterization of cyclic AMP-mediated wound closure of the rabbit corneal epithelium. PMID- 6277569 TI - Na+ transport across the rabbit corneal endothelium. AB - It has been reported so far that the Na+ transport across the corneal endothelium is symmetrical and therefore, that there is no net transport. We now report that there is a net Na+ transport across the rabbit corneal endothelium. Instead of measuring the conventional steady-state fluxes, pre-steady state fluxes were measured as a function of time (readings were taken at two minute intervals). This technique is based on a theory, proposed very recently, that the flux ratio (efflux/influx) is independent of time. Both the efflux and influx reached their steady state values about fifteen minutes after the addition of the isotope to one side of the chamber. The net sodium flux was obtained from the difference between the steady state efflux and influx values. The direction of the net Na+ flux was from the stromal to the aqueous side, with a magnitude of 2.3 +/- 0.4 microEq/h.cm2 (n = 11, S.E.M.). The net Na+ transport was inhibited by the presence of ouabain (10(-4)M). PMID- 6277570 TI - The effects of glutathione and adenosine on plasma membrane ATPases of the corneal endothelium. An hypothesis on the stimulatory mechanism of perfused glutathione upon deturgescence. AB - Reduced glutathione (0.3 mM) stimulates the activity of sodium-potassium activated ATPase (Na+K+ATPase) by 54% in plasma membranes prepared from bovine corneal endothelial cells. Oxidized glutathione, however, has no effect on Na+K+ATPase activity in the same tissue, although it does inhibit magnesium activated ATPase (Mg++ATPase) by approximately 30%. Adenosine neither stimulates nor inhibits either Na+K+ATPase or Mg++ATPase in these plasma membranes. It is postulated that the stimulatory effect of glutathione on deturgescence stems from the direct reaction of the reduced form of the tripeptide on sulfhydryl groups located on plasma membranes of corneal endothelial cells. It is highly probable that these sulfhydryl groups are part of the Na+k+ATPase complex itself. PMID- 6277571 TI - Regulation of the histidine operon: translation-controlled transcription termination (a mechanism common to several biosynthetic operons). PMID- 6277572 TI - In vivo functioning of the Na+, K+-activated ATPase. PMID- 6277573 TI - High resolution computerized tomography with coronal and sagittal reconstruction in the diagnosis of brain tumors in children. AB - We present 4 children with brain tumors that were missed or incompletely characterized conventional computerized tomography (CT) scanning. Using high resolution CT with high dose contrast and three-dimensional reconstruction, we were able to define the anatomy of each tumor and its associated aberrant vasculature. High resolution CT scanning with reconstruction is very helpful in diagnostic problems in which conventional CT scanning fails to provide a clear diagnosis. PMID- 6277574 TI - [Surgical tactics in precancerous conditions and non-invasive cancers of the breast]. AB - In the treatment of non-palpable lesions of the breast, geometric localization technique with subcutaneous segmental mastectomy and intraoperative specimen radiography proved to be safe. Permanent sectioning of the whole excised area is recommended to improve the reliability of diagnosis. If breast biopsy establishes proliferative mastopathy with cell atypia or papillary proliferation, a strict clinical follow-up of the patient is adequate. When lobular neoplasia is detected, both breasts are in danger of developing carcinoma. In the treatment of this lesion, the calculated risk of life-long follow-ups may sometimes be an acceptable alternative to the certain cure by (subcutaneous) mastectomy. Yet, for treatment purposes intraductal carcinoma, with its risk of occult infiltration, should be considered potentially invasive. PMID- 6277575 TI - [Metastatic cystosarcoma phyllodes]. PMID- 6277576 TI - [A preliminary study on the relationship between cAMP and hemopoietic function (author's transl)]. PMID- 6277577 TI - [Investigation of cyclic adenosine monophosphate (cAMP) of the blood, urine and CSF in schizophrenics (author's transl)]. PMID- 6277578 TI - [A preliminary report on the indicator of the enterovirus in hospital sewage (comparative studies on the resistance of f2 coliphage, B. cereus spore and HBsAg to chemical agents) (author's transl)]. PMID- 6277579 TI - Etiopathogenesis of excess methylprednisolone arterial hypertension in the rat. AB - The blood pressure response to different doses of methylprednisolone was examined in the rat. It is concluded that doses varying from 2.5 mg/kg/week to 20 mg/kg/week of this agent caused clear-cut elevations in arterial pressure. The methylprednisolone-induced arterial hypertension was accompanied by elevation in Plasma Renin Activity and administration of captopril or saralasin caused significant drops in systemic arterial pressure. Concomitant long term administration of captopril and methylprednisolone caused a delay in appearance and smaller elevations in arterial pressure. It is concluded the methylprednisolone in the rat causes arterial hypertension which is at least partially dependent upon renin angiotensin system activation. However elevated blood pressure levels were noticeable even during chronic captopril administration leading to the conclusion that other mechanism (s) may participate in the pathogenesis of this experimental model of hypertension in rats. PMID- 6277580 TI - [Molecular cloning of DNA fragments homologous to mouse sarcoma virus oncogen from a collection of genes from transformed cells]. PMID- 6277581 TI - [Ion channel coupling with neutral amino acid receptors in the endings of rat spinal afferents]. PMID- 6277582 TI - [Analysis of the acetylcholine "noises" recorded in fast and slow muscle fibers of the frog]. PMID- 6277583 TI - [Cloning and restriction analysis of the hepatitis B virus genome]. PMID- 6277584 TI - [Organization of the genetic system of plasmid R6K conjugativity]. PMID- 6277585 TI - [Action of cyclic nucleotides on the cytoplasmic membrane conductance of the rod outer segment in the retina]. PMID- 6277586 TI - [Interaction with liposomes of the spin-labelled cytotoxin from Central Asian cobra venom]. PMID- 6277587 TI - Captopril - a genuine innovation. PMID- 6277588 TI - [Therapy of esophageal reflux disease]. PMID- 6277589 TI - [Chemotherapy of listeriosis (author's transl)]. AB - Despite the occasional lack of antibacterial activity ampicillin is considered to be the drug of choice for antibacterial chemotherapy in listeriosis. In vitro comparison with new penicillins (mezlocillin, piperacillin) and cephalosporins (cefamandol, cefoxitin, cefuroxime, cefotaxime) showed ampicillin to have the most potent activity against Listeria monocytogenes. Minimal inhibition concentrations (MIC) were between 0.06 and 1 microgram/ml and minimal bactericidal concentrations (MBC) from 0.25 to 32 micrograms/ml. The MIC90 (minimal concentration inhibiting 90% of tested strains) was 0.48 for ampicillin, 8.0 for mezlocillin and 5.2 micrograms/ml for piperacillin. Among cephalosporins cefalotin was most effective with an MIC90 of 5.8 micrograms/ml. The MIC90 was 7.8 for cefamandol and 89.6 micrograms/ml for cefoxitin. Cefuroxime and ecfotoxime had MIC values of more than 128 micrograms/ml and showed no clinically relevant anti-listeria activity. Gentamicin and doxycyclin showed MIC90 values of 12 and 8 micrograms/ml, respectively. MBC and MIC values were closest together in gentamicin. Ampicillin, potentially combined with gentamicin, should remain treatment of choice in generalised Listeria monocytogenes infection. PMID- 6277590 TI - [Extrahepatic manifestations of acute viral hepatitis]. PMID- 6277591 TI - [Steroid hormone receptors in breast carcinoma]. PMID- 6277592 TI - [The influence of several membrane stabilizing drugs on the induced hereditary myotonia--a methodic way to test the effectiveness of drugs on myotonia (author's transl)]. AB - The therapeutic effectiveness of several membrane stabilizing drugs was investigated on experimentally induced myotonia by 2,4-Dichlorophenoxyacetate (2,4-D) and in 20 patients with myotonia congenita. Procainamide and quinine showed a better antimyotonic effect in-vitro- as well as in-vivo-experiments on the cold blood muscle and the rat than Sparteine sulfate and Tachmaline (Ajmalin). The two last-mentioned drugs had nearly the same effect. Because of these experimental results only procainamide and sparteine sulfate were clinically used. Quinine was not used because of the well known side-effects. Mention is made of the fact, that the therapeutic effect depends on the dose or the concentration. The results support not only the theoretic considerations on the pathogenesis of myotonia but also recommend to carry out further pharmacological investigations with this method. PMID- 6277593 TI - Regulation of cAMP-dissociation kinetics in lactating rat mammary gland. AB - The cAMP-dissociation kinetics of rat mammary gland cytosols are dependent upon the temperature of cAMP association. Dissociation rates (measured at pH 6.5, 24 degrees C) were biphasic (k = 0.08-0.23 min-1 and k = 0.02 min-1) and monophasic (k-1 = 0.02 min-1) after 0 degrees C and 24 degrees C association, respectively. The temperature-dependent change from an initial fast rate to an initial slow rate was observed at all concentrations of cAMP tested from 1 to 1000 nM. When the slow-dissociating site was associated with non-radioactive 8-bromo-cAMP, the dissociation rates of [3H]-cAMP from the remaining dissociating site was slow (k = 0.02 min-1) and fast (k = 0.05 min-1) at 24 degrees C and 0 degrees C associating rate can be converted to the slow-dissociating rate by warming. When 0.2 M sodium thiocyanate was added to the association mixture at 24 degrees C, biphasic dissociation rates of k = 0.23 min-1 and k = 0.02 min-1 were observed, suggesting that the chaotropic salt blocks the interconversion of rates. The data are consistent with the model for cAMP-dependent protein kinase which exhibits two binding sites with different affinities. The type II enzyme from mammary gland cytosol exhibits in addition the phenomenon of temperature-dependent interconversion of the two binding affinities. PMID- 6277594 TI - Hypothalamic regulation of pituitary gonadotropin-releasing hormone receptors: effects of hypothalamic lesions and a gonadotropin-releasing hormone antagonist. PMID- 6277595 TI - Hypothalamic regulation of pituitary gonadotropin-releasing hormone receptors: effects of gonadotropin-releasing hormone immunoneutralization. PMID- 6277596 TI - Suckling withdrawal increases pituitary lysosomal enzyme activities and prolactin protease in lactating rats. PMID- 6277597 TI - Specific inhibition by glucocorticoids of the alpha 2-adrenergic stimulation of adrenocorticotropin release in rat anterior pituitary cells. AB - Previous studies have shown that a specific alpha 1-adrenergic receptor leads to a 7- to 12-fold stimulation of ACTH release in rat adenohypophyseal cells in culture. The ACTH response to epinephrine is inhibited by 65% after a 3-h incubation with the glucocorticoid agonist dexamethasone, whereas a complete suppression of the response is observed when the steroid is added 4 h earlier. Upon placing the inhibited cells in steroid-free medium, a 50% reversal of the inhibition of the ACTH response is observed after 10 h, whereas normal responsiveness is obtained after 36 h. Glucocorticoid agonists lead to a complete inhibition of epinephrine-induced ACTH secretion with the following order of potencies (ED50 values): triamcinolone acetonide (0.2 nM) much greater than dexamethasone (1.5 nM) much greater than cortisol (11 nM) greater than corticosterone (22 nM). Preincubation with dexamethasone for 4 h leads to an 85% decrease in the maximal ACTH response to epinephrine, whereas the ED50 value of catecholamine action is only slightly increased. Although the activity of the pituitary-adrenocortical axis is highly sex dependent in the rat, 17 beta estradiol, 5 alpha-dihydrotestosterone, and the pure progestin R5020 have no detectable effect on either basal or epinephrine-induced ACTH release, thus suggesting that sex steroids exert their action in the intact animal at sites other than in corticotrophs. The finding of a specific inhibitory effect of glucocorticoids on epinephrine-induced ACTH secretion lends further support to the suggestion of a physiological role for the pituitary alpha 1-adrenergic receptor in the control of ACTH secretion (at least in the rat). The complete lack of effect of sex steroids indicates the high degree of specificity of the feedback mechanisms controlling ACTH secretion. PMID- 6277598 TI - Multiplication-stimulating activity stimulates the multiplication of F9 embryonal carcinoma cells. AB - The present study was designed to assess the effectiveness of multiplication stimulating activity (MSA), an insulin-like growth factor, in supporting the growth of F9 cells (an embryonal carcinoma line with properties similar to embryonic stem cells). Under serum-free growth conditions in a medium supplemented with transferrin and fibronectin, MSA is an effective mitogen. At 100 ng/ml, MSA completely replaces the growth requirement for fetal calf serum. Biologically active MSA polypeptides II and III act as potent growth promoters of F9 cells, whereas MSA I appears to be somewhat less effective. Binding studies carried out with [125I]iodo-MSA indicate that F9 cells possess specific receptors for MSA. Scatchard analysis indicates a single class of MSA receptors with a Ka of 8.2 x 10(9) M-1; about 55,000 MSA molecules can bind per F9 cell. Insulin-like growth factor I and II both complete for MSA binding, whereas insulin does not compete. Since insulin is an effective promoter of F9 cell growth, these results indicate that the mitogenic action of insulin toward F9 cells is not mediated through MSA receptors. It is apparent from these results that MSA is capable of serving as an early embryonic growth factor. PMID- 6277599 TI - Quinine and the stimulus-secretion coupling in pancreatic beta-cells: glucose like effects on potassium permeability and insulin release. PMID- 6277600 TI - Demonstration and organ distribution of the 1,25-dihydroxyvitamin D3-binding protein in fish (A. anguilla). PMID- 6277601 TI - Steroidogenic capacity of isolated adult mouse Leydig cells does not decrease with age. AB - Leydig cells were isolated from the testes of adult young (5--7 months), old (21 months), and senile (27 months) mice using a highly preservative Percoll procedure. The yield in Leydig cells per testis from young animals was slightly lower than from the old age groups. hCG-induced testosterone synthesis proceeded in a linear fashion for at least 5 h in all three groups. The maximal steroidogenic capacity of cells from old animals was identical to that from young adults. There was no significant difference between Leydig cells from young and old mice with respect to hCG-induced cAMP accumulation and protein kinase activation. Determination of hCG concentrations required for half-maximal stimulation of testosterone synthesis and cAMP accumulation showed identical, or even lower, values in the old age groups. The phenomenon may be connected with the significant augmentation with age of the DNA content per cell (polyploidization), possibly acting as a compensatory mechanism of age-induced deficiencies. Detailed kinetic studies of cAMP accumulation, protein kinase activation, protein kinase activation, and steroidogenesis as well as ultrastructural analyses support the findings of unimpaired or increased capacities of the testosterone-forming cells in old animals. Thus, the aging of Leydig cells appears to differ from that of other tissues of the mouse (e.g. skeletal muscle), which exhibit decreasing abilities to respond to stimuli. PMID- 6277602 TI - Adenosine 3',5'-monophosphate-dependent protein kinase in supernatants from dispersed bovine parathyroid cells. AB - The presence and characteristics of protein kinase(s) were studied in supernatants of sonicates of dispersed bovine parathyroid cells. cAMP caused a 3- to 5-fold stimulation of protein kinase activity in such extracts, with half of the maximal activation at 4--5 x 10(-8) M cAMP. Protein kinase inhibitor nearly totally abolished both basal and cAMP-stimulated activity, suggesting that most of the activity was cAMP dependent. About 90% of the cAMP-stimulated protein kinase activity eluted from a DEAE-cellulose column at 0.15 m NaCl, consistent with a type II enzyme. The presence of a type II enzyme was also supported by the effects of histone and salt concentrations on enzyme activity; both the basal and cAMP-stimulated activity ratios (activity minus cAMP divided by activity plus 10( 6) M cAMP) were stable in 0.4 M NaCl. The basal activity ratio was not increased by concentrations of histone as high as 10 mg/ml in the protein kinase assay. The predominance of the type II enzyme in dispersed bovine parathyroid cells made it possible to develop conditions for extracting the enzyme from intact intact cells (0.4 m NaCl and 5 mg/ml charcoal), whereby the state of activation of the enzyme remained relatively constant. These studies demonstrate the presence of cAMP dependent protein kinase activity in dispersed bovine parathyroid cells and define conditions which make it possible to assess the effects of various secretagogues on protein kinase activation in intact parathyroid cells. PMID- 6277603 TI - Time trend analysis of plasma unconjugated and sulfoconjugated estrone and 3 beta delta 5-steroids in fetal and maternal sheep plasma in relation to spontaneous parturition at term. AB - Parturition in the sheep is preceded by a complex series of changes in both fetal and maternal plasma-steroid hormone concentrations. Using the chronically catheterized fetal sheep preparation, we measured unconjugated and sulfoconjugated pregnenolone, 17 alpha-hydroxypregnenolone, dehydroepiandrosterone, and estrone in fetal and maternal plasma over the final 20 days before spontaneous vaginal delivery at term. Where appropriate, third degree polynomial functions were fitted to the changing plasma hormone concentration profile. Fetal and maternal plasma pregnenolone and pregnenolone sulfate both fell from maximum values in the last 4 days of gestation. Fetal and maternal plasma estrone and estrone sulfate concentrations underwent a terminal rise over the last 4 days of gestation that was a mirror image of the fall in plasma pregnenolone and pregnenolone sulfate. Maternal 17 alpha hydroxypregnenolone rose over the last 4 days of gestation. Fetal 17 alpha hydroxypregnenolone, maternal and fetal plasma dehydroepiandrosterone sulfate, and fetal plasma dehydroepiandrosterone sulfate, and fetal plasma dehydroepiandrosterone showed no trend during the period of study. Maternal plasma dehydroepiandrosterone rose over the last 4 days of gestation. These results support the view that increased activity of placental 17 alpha hydroxylase and 17-20-desmolase is responsible for the conversion of C-21 steroids to estrogens at term. delta 5-Steroids are present in very high plasma concentrations in fetal sheep plasma and may constitute a more important precursor pool for estrogen biosynthesis than does circulating plasma progesterone. PMID- 6277604 TI - Induction of parturition in sheep by low dose intrafetal infusion of synthetic adrenocorticotropin-(1-24) at 120-130 days gestation. PMID- 6277605 TI - Apparent dissociation of adrenocorticotropin and corticosteroid responses to 15 ml/kg hemorrhage in conscious dogs. AB - The blood pressure, heart rate, ACTH, corticosteroid, vasopressin, and renin responses to rapid 15 ml/kg hemorrhage were measured in six conscious healthy dogs with chronically maintained femoral arterial catheters. The hemorrhage decreased the mean arterial blood pressure slightly (P less than 0.001), increased the heart rate (P less than 0.001), and increased arterial plasma levels of ACTH (P less than 0.01), corticosteroids (P less than 0.001), vasopressin (P less than 0.001), and renin activity (P less than 0.001). Overall and in the individual experiments, there appeared to be little correspondence between the ACTH and corticosteroid responses. In none of the experiments was there a clear rise in ACTH above control levels before the first rise in corticosteroids. To ascertain that adrenal secretion of corticosteroids was increased during 15 ml/kg hypovolemia, changes in the clearance and distribution volume of cortisol were estimated by counting tritium extracted from plasma of five dogs infused with [1,2-3H] cortisol to steady state levels before and during hypovolemia. The stimulus caused a 30% reduction from steady state levels of dichloromethane-extractable tritium counts (P less than 0.001). Combined with the observed increase in plasma corticosteroid levels, these results show that the increase in adrenal secretion of corticosteroids after hemorrhage was underestimated by measurement of changes in peripheral plasma levels. The hypothesis that hemorrhage results in an increase in adrenal sensitivity to ACTH is tested in the following paper. PMID- 6277606 TI - Adrenal sensitivity to adrenocorticotropin in normovolemic and hypovolemic conscious dogs. PMID- 6277607 TI - Equine Cushing's disease: plasma immunoreactive proopiolipomelanocortin peptide and cortisol levels basally and in response to diagnostic tests. PMID- 6277608 TI - In vitro effects of ethanol on primate luteal membranes: exposure of additional gonadotropin receptor sites. AB - The presence of 0.8-8.0% (vol/vol) ethanol increased the specific binding of [125I]human luteinizing hormone (hLH) to membrane particulate fractions prepared from the rhesus monkey corpus luteum in a dose-dependent manner. Ethanol increased the number of available gonadotropin binding sites without altering their affinity for LH. These results suggest that (1) there are additional gonadotropin receptors in the macaque corpus luteum which are not detected by conventional in vitro techniques, and (2) ethanol exposes receptors previously unavailable for hormone interaction. PMID- 6277609 TI - Simple goiter and its variants: euthyroid and hyperthyroid multinodular goiters. PMID- 6277610 TI - Relaxin as a new hormone. PMID- 6277611 TI - Concerning the coidentity of phosphatidic acid phosphohydrolase and phosphatidylglycerophosphate phosphohydrolase in rat lung lamellar bodies. AB - The properties of the phosphatidylglycerophosphate phosphohydrolase activity in lamellar bodies from rat lung have been compared with the properties of the activities responsible for the degradation of aqueously-dispersed phosphatidic acid (PAaq) and membrane-bound phosphatidic acid (PAmb). Subcellular fractionation studies revealed that the phosphatidylglycerophosphate phosphohydrolase activity and the PAaq-dependent phosphatidic acid phosphohydrolase activity were predominantly associated with the mitochondrial and microsomal fractions, while the PAmb-dependent phosphohydrolase activity was associated with the cytosol. Although the lamellar body fraction contained less than 1% of the total activity, the phosphatidic acid phosphohydrolase activities associated with this fraction could not be explained by contamination with microsomes or cytosol. The three activities exhibited similar heat inactivation profiles at 55 degree C. However, differences in the responses of these activities to the presence of iodoacetate, p-chloromercuriphenyl sulphonate, mercaptoethanol, mercaptoethanol plus MgCl2, and Triton X-100 indicated that the enzymes responsible for these activities may be distinct. Furthermore, addition of up to a ten-fold greater amount of PAaq did not seriously affect the hydrolysis of phosphatidylglycerophosphate. These results indicate that the phosphatidic acid phosphohydrolase and phosphatidylglycerophosphate phosphohydrolase activities in rat lung lamellar bodies are not necessarily catalyzed by the same protein. PMID- 6277612 TI - Lung phosphatidate phosphatase: activity during altered physiologic states. AB - Phosphatidic acid phosphatase (PAPase) which catalyzes the conversion of phosphatidic acid to 1,2-diacyl-sn-glycerol was studied in fetal, neonatal, and adult rat lung microsomal fractions from whole lung under normal and altered physiological states. The maximal activity was obtained at pH 7.0 with 1.0 mM phosphatidic acid as the substrate. Twenty-one-day-old fetal rat lung averaged 20.3 +/- 0.6 SE nmol/min/mg microsomal protein compared to 9.9 +/- l.0 nmol/min/mg in liver. Following birth there was a dramatic 53% increase in the PAPase activity. Twenty-one-day-old fetal rat lungs from diabetic mothers (streptozotocin-induced) and from mothers fasted the last four days of gestation did not show altered PAPase activity. Premature breathing for 3-6 hr on day 21 of gestation also did not affect the PAPase activity. These data demonstrate that the microsomal PAPase activity (l) increases dramatically only after birth (2) is not responsive to altered physiologic state (maternal diabetes, maternal fasting, and premature breathing) and (3) may not be an important regulatory enzyme in lung surfactant phospholipid production. PMID- 6277613 TI - Excess lung cancer risk in a synthetic chemicals plant. AB - A standardized mortality ratio of 1.49 for respiratory system cancer (42 observed deaths versus 28.2 expected, p less than 0.01) was observed among a cohort of 4806 males employed at a synthetic chemicals plant since its startup in 1942. Upon review of pathologic material, the excess was found to be limited to adenocarcinoma and large cell undifferentiated lung cancer. Many of the workers had been exposed to vinyl chloride, as well as to chlorinated solvents, poly(vinyl chloride) (PVC) dust, acrylates and acrylonitrile. To evaluate the association between lung cancer and occupational chemical exposures, detailed work histories for each cohort member were combined with exposure ratings for each of 19 chemicals for each job for each calendar year since 1942. A serially additive expected dose model was then constructed which compared the doses of the chemicals observed for the lung cancer cases to the doses expected based on subcohorts without lung cancer individually matched to the cases. PVC dust appeared to be the most likely etiologic agent (p = 0.037). Time trends of PVC dust exposure indicated a potential latent period of 5-16 years before death. PMID- 6277614 TI - Effect of ethanol on vinyl chloride carcinogenesis. AB - Four treatment groups (80 male Sprague-Dawley rats/group) were used in a 2 X 2 factorial design: inhalation of 600 ppm vinyl chloride (VC) 4 hr/day, 5 days/week for 1 year; VC and ingestion of 5% ethanol in water (v/v); filtered air and ethanol; filtered air. Ingestion of ethanol was begun 4 weeks prior to inhalation of VC and continued for life or termination of the study at two and one-half years from the first VC exposure. In this model system, ethanol potentiated the carcinogenic response to VC in the liver and produced an excess of neoplasms in animals receiving ethanol alone. Inhalation of VC induced angiosarcoma of the liver in 23% of the exposed animals; ethanol in addition to VC inhalation increased the incidence to 50%. Concomitant administration of VC and ethanol also produced an excess of hepatocellular carcinoma and lymphosarcoma. Ethanol with or without VC had a strong tumorigenic effect on the endocrine system. These results indicate that ethanol is a cocarcinogen in relation to the carcinogen VC. PMID- 6277615 TI - Carcinogenicity of airborne fine particulate benzo(a)pyrene: an appraisal of the evidence and the need for control. AB - Benzo(a)pyrene(BaP) originating from fossil fuel and other organic combustion processes is largely adsorbed on fine particulate and hence is a widespread atmospheric pollutant. Available emissions and air quality data are based on the total weight of particulate matter without reference to size and give little information on trends and concentrations of fine particulate BaP. Greater reliance on coal, synfuels and diesel fuel for energy production and transportation will significantly increase ambient levels of BaP. Because of the particulate size, BaP is substantially deposited in the lower lung and readily eluted into surrounding tissue. After elution in the lung, BaP is metabolically activated to its electrophilic, carcinogenic from by a complex enzyme system whose activity is increased by prior exposure to air pollutants, cigarette smoke and certain drugs. The resultant diol epoxide metabolite has been shown to bind covalently with the DNA of the lung. In experimental animals, BaP is a potent initiating carcinogen whose action is enhanced by sulfur dioxide, promoting agents and carrier fine particles. The effect of small, divided doses of BaP has been shown to be greater than that of a single high dose; no threshold has been established. Epidemiological studies show that mixtures containing BaP (such as urban air, industrial emissions and cigarette smoke) are carcinogenic and may interact synergistically. Occupational studies indicate that the action of BaP containing mixtures is enhanced in the presence of SO2. However, quantitative risk assessment for BaP is precluded by problems in extrapolating to the general population from small-scale animal studies; uncertainties in findings of epidemiology; and imprecise exposure data. Existing stationary and mobile controls preferentially remove coarse particulate matter and are inefficient collectors of the particulate BaP. In the current absence of health and environmental standards for BaP, there is little incentive to control BaP emissions. BaP meets the criteria for regulation under the Clean Air Act; however, no such BaP standards have yet been proposed. PMID- 6277616 TI - Assessment of myelotoxicity caused by environmental chemicals. AB - Potential antineoplastic agents must be screened for the delayed toxicity that occurs in many cases of drug-induced bone marrow aplasia. In vitro clonal assays for hematopoietic progenitor cells have been developed to assess the degree of myelotoxicity. This adverse side effect is often the limiting factor in the development of new cancer chemotherapeutics. In addition, many environmental chemicals are cytotoxic to rapidly proliferating cells, but a systematic assessment of their myelotoxicity has not been performed. We have used clonal marrow assays to investigate a panel of chemicals including 2,3,7,8 tetrachlorodibenzo-p-dioxin, polybrominated biphenyls, diethylstilbestrol, benzo(a)pyrene and indomethacin. All were immunotoxic, some to pleuripotent hemopoetic stem cells and other to granulocyte-macrophage progenitors, and at concentrations below those causing other toxic manifestations. This shows that these bone marrow clonal assays, and hopefully future one for erythroid, B- and T lymphocytes, and megakaryocytes, will provide the specificity and sensitivity necessary to delineate the myelotoxicity of a broad spectrum of environmental chemicals. PMID- 6277618 TI - Effects of tunicamycin on the cell-surface binding, internalization and degradation of low-density lipoproteins in human fibroblasts. PMID- 6277617 TI - Approaches to assessing host resistance. AB - There is increasing evidence that chronic, subclinical exposure to certain environmental pollutants may upset immune responsiveness and alter susceptibility of animals to infectious agents. Environmental chemicals or drugs may affect diverse aspects of the immune system, leading to immunosuppression, immunopotentiation, hypersensitivity or perturbed innate host resistance. A variety of infectious models is available that involves relatively well defined target organs and host defense mechanisms; for example, infections with encephalomyocarditis virus, Herpesvirus simplex, Listeria monocytogenes, Streptococcus pneumoniae, Escherichia coli or Plasmodium berghei. Important variables in infectious models used to assess immunotoxicity include species and strain of animal used, their age and sex, the route of exposure, and dose of the chemical. No one infectious model has yet emerged as a routine screening tool to detect and assess the subtle effects that may occur in immune responses when animals are exposed to doses of environmental pollutants that cause no adverse effect at a gross level. The selection of useful test systems is complicated because it is difficult to measure the effects of chronic, subclinical exposure to chemicals and sublethal challenges of microorganisms. PMID- 6277619 TI - Purification and properties of an endonuclease from the mitochondrion of Saccharomyces cerevisiae. AB - An endonuclease, which is found only in the mitochondrion of the yeast Saccharomyces cerevisiae, has been purified. The protein has a sedimentation coefficient of 6.3 S, equivalent to a molecular weight of 105,000. The enzyme is active at pH 7.6, when it degrades single-stranded DNA about 10-times faster than double-stranded DNA, but at pH 5.4 only double-stranded DNA is degraded. In both cases the enzyme acts endonucleolytically, breaking a single phosphodiester bond at a random location within the DNA substrate. Mn2+ or Mg2+ are required for activity; Ca2+ and Zn2+ are ineffective cofactors. Enzyme activity at pH 7.6 is severely inhibited by low concentrations of NaCl or KCl, while activity at pH 5.4 is unaffected by salt. Ethidium bromide inhibits both the DNase activity at pH 5.4 and the activity with single-stranded DNA at pH 7.6, but has no effect on the DNase activity with double-stranded DNA at pH 7.6. PMID- 6277620 TI - Dioxygen activation by putidamonooxin. The oxygen species formed and released under uncoupling conditions. AB - In the presence of substrates not favourable for hydroxylation, more than 80% of the dioxygen consumed by purified, reconstituted 4-methoxybenzoate monooxygenase appears in the reaction mixture as hydrogen peroxide. We have investigated whether under these conditions (a) reduced putidamonooxin, the oxygenase of this enzyme system, either autoxidizes in the presence of dioxygen, with liberation of superoxide anion radicals which then disproportionate to H2O2 and O2, or (b) dioxygen is reduced by two sequential single-electron steps leading to the active oxygen species that forms hydrogen peroxide directly when inactivated by protonation. Quantitative estimation of O-2 radicals, with either succinylated ferricytochrome c or epinephrine used as O-2 scavengers, revealed that only about 6% of the total electron flux channelled via putidamonooxin to dioxygen led to the monovalent reduction on dioxygen. This means that not more than 3% of the hydrogen peroxide found under uncoupling conditions arises from the rapid bimolecular disproportionation of initially formed O-2 radicals. Inconsistent results were obtained when lactoperoxidase was used as an O-2 trap. Our measurements indicate that the conversion of lactoperoxidase into compound III is an inappropriate method of detecting any O-2 radicals that may be found by the uncoupled 4-methoxybenzoate monooxygenase. The stoichiometry of about 1:1 for O2 uptake: H2O2 formation indicates that under uncoupling conditions H2O is virtually not formed. The role of [FeO2]+ as the active oxygenating species of putidamonooxin is discussed. PMID- 6277621 TI - Control of ketogenesis and fatty-acid synthesis at the mitochondrial branch-point for acetyl-CoA in the chick liver cell: effect of adenosine 3',5'-monophosphate. PMID- 6277622 TI - R-loop mapping of calf-thymus RNA polymerase II. Transcripts in vitro on restriction fragments of adenovirus 2 DNA. AB - Nascent RNA, synthesized by calf thymus ENA polymerase II on the restriction endonuclease EcoRI fragment A and BamHI fragment B of adenovirus 2 DNA, was rehybridized to the template strand under conditions allowing transcription R loop formation. Hybrids, visualized by electron microscopy, were found in looped and in branched configurations, the former being abundant, with an average loop number of roughly four per EcoRI fragment A and three per BamHI fragment B. Frequency distributions of transcription R-loops, synthesized on identical restriction fragments for different periods of time, showed similar patterns, thus pointing to a nonrandom type of transcription. A comparison of the frequency distribution of R-loops seen on the two overlapping restriction fragments reveals reasonable coincidence in the loop pattern in one out of the four possible fragment orientations, permitting correlation of the transcription R-loop pattern with the physical map of the adenovirus 2 DNA. Comparison of start sites tentatively mapped in vitro with the known promoters for RNA polymerase II in vivo suggests that a major transcription site in vitro is located in close proximity to, or identical with, two promoters in vivo at 16.1 and 16.5 map units. PMID- 6277623 TI - Alterations induced by a prolonged fasting: opposite effects on the beta adrenergic receptor-coupled adenylate-cyclase system and on lipolysis in fat cells from rat. PMID- 6277624 TI - DNA fragment conformations. 1H-NMR and relaxation studies of 2'-deoxyadenylyl(3' 5')thymidylyl-(3'-5')deoxyguanosylyl(3'-5')thymidine, d(A-T-G-T), in neutral aqueous solution. PMID- 6277625 TI - The mechanisms of action of cAMP. A quantum chemical study. AB - Quantum chemical calculations were performed on the formation of intermediates with trigonal bipyramidal (TBP) configurations in the hydrolysis of adenosine 3',5'-monophosphate (cAMP) with phosphodiesterases and the activation of protein kinases by cAMP. The results show that in the reaction sequence concerning the hydrolysis of cAMP with phosphodiesterase the TBP intermediate must possess an equatorial-apical cyclic phosphate ring with the 3'-oxygen atom in the apical position. This could be an additional reason for the sensitivity of the 3' position in cAMP towards modifications in comparison with the 5' position. According to the calculations, a mechanistic model is presented for the enzymatic hydrolysis of cAMP with the involvement of a covalently bonded enzyme-nucleotide intermediate. Also a model is offered for the activation of protein kinase by cAMP. The activation of protein kinase is assumed to proceed via diequatorial ring-positioned TBP intermediates resulting in the formation of a covalent bond between cAMP and the protein kinase with retention of the cyclic phosphate ring. It seems likely that the enzyme-nucleotide intermediate enforces a conformational change in the enzyme, which causes the dissociation of the regulatory and catalytic subunit of the protein kinase, necessary for a physiological response. PMID- 6277626 TI - Incorporation of deoxyribonucleosides into DNA of coryneform bacteria and the relevance of deoxyribonucleoside kinases. AB - In order to obtain basic knowledge of the salvage pathways for DNA synthesis, the ability of Brevibacterium ammoniagenes ATCC 6872 and Micrococcus luteus ATCC 15932 for incorporation of nucleobases and nucleosides was investigated. Only adenine and uracil are incorporated by B. ammoniagenes, whereas M. luteus additionally can utilize deoxyadenosine and, less efficiently, thymidine. In M. luteus, the demonstration of deoxyadenosine kinase and thymidine kinase explains the incorporation data. Uptake of thymidine is of short duration because of rapid breakdown of exogenously supplied thymidine to thymine. At a 540-fold excess pyrimidine deoxyribonucleosides inhibit 14C incorporation from thymidine nearly totally and purine deoxyribonucleosides cut by half the uptake rate, probably by interfering with transport of thymidine. However, as no cessation of thymidine incorporation occurs at these concentrations of purine deoxyribonucleosides, incorporation is finally enhanced. During the initial period of this reduced uptake considerable protection of thymidine from breakdown to thymine is provided by deoxyguanosine, but not by deoxyadenosine. At a 108-fold excess there is actually no inhibition of thymidine uptake by deoxyguanosine and only an insignificant impairment by deoxyadenosine resulting in an ultimate enhancement of 14C incorporation up to 20% of the exogenously supplied thymidine. As there is no salvage pathway for thymidine in B. ammoniagenes due to the absence of thymidine kinase, labelling with adenine and hydrolyzing of the 'contaminated' RNA fraction with 1 M KOH is recommended for measurements of overall DNA synthesis in this strain. PMID- 6277627 TI - Isolation and some properties of the restriction endonuclease BcnI from Bacillus centrosporus. AB - A specific type-II restriction endonuclease BcnI from Bacillus centrosporus has been purified to electrophoretic homogeneity in three chromatographic steps. Around 15 micrograms of such a preparation can be isolated from 1 g of the cell paste. The yield of the enzyme is higher than that of any type-II restriction endonuclease so far reported. The molecular weight of the enzyme determined by gel filtration and polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulphate equals 27 500 and 28 000 respectively. The activity of the restriction endonuclease is maximal at pH 9.2 and 40--45 degrees C. The optimal magnesium concentration was estimated to be 7.5 mM. The activity of BcnI may also be observed in the presence of Co2+, Mn2+, Ni2+ and Zn2+ but it is markedly less than in the presence of Mg2+. PMID- 6277628 TI - Modification of the rapidly reacting thiols of atrial and ventricular myosins. Evidence for structural variances around the SH-1 thiol of the two isozymes. AB - Environmental variances were noted around the rapidly reacting SH-1 thiol of ventricular versus atrial myosin, based on electron paramagnetic resonance studies. Further studies, in which either SH-1 or SH-1 + SH-2 thiols were modified with N-ethylmaleimide, indicated the importance of the SH-2 moiety of both isozymes for generation of tension, when analyzed as synthetic actomyosin threads. Comparative EPR studies showed that the spin label was more strongly immobilized when complexed to ventricular SH-1 thiol as compared to when it was complexed to atrial myosin. Likewise, addition of PPi, ATP or ADP created a greater mobility in the spin label when added to ventricular spin-labeled myosin as compared to that of atrial myosin. comparative studies of spin-labeled actomyosin versus myosin analyzed at different EPR power settings also demonstrated disparaties surrounding the SH-1 thiol between the two myosin isozymes. PMID- 6277629 TI - Biosynthesis of pairs of peptides related to melanotropin, corticotropin and endorphin in the pars intermedia of the amphibian pituitary gland. AB - This study concerns the biosynthesis of a number of peptides in the neurointermediate lobe of the pituitary gland of the aquatic toad, Xenopus laevis. Using pulse-chase incubations in vitro and high-performance liquid chromatographic analysis, it could be shown that these peptides are synthesized through processing of a prohormone, pro-opiomelanocortin; all peptides were released into the incubation medium. On the basis of electrophoretic analysis, selective amino acid incorporation and immunoprecipitation, as well as peptide mapping by high-performance liquid chromatography, the peptides were classified into three distinct groups: two related to melanocyte-stimulating hormone (melanotropin), two related to adrenocorticotropic hormone (corticotropin) and two endorphin-like peptides. Using tryptic and chymotryptic maps of synthetic alpha-melanotropin and des-Ac alpha N-alpha-melanotropin as references, one of the melanotropin-like peptides was identified as des-Ac alpha N-alpha melanotropin; the other one represents neither alpha-melanotropin nor any other known melanotropic peptide. The two peptides that were immunologically related to corticotropin had characteristics consistent with a structures resembling a peptide previously named 'corticotropin-like intermediate lobe peptide', corticotropin-(18-39). The two endorphin-like peptides, although highly related, do not have the same primary structure. In view of the apparent structural differences between the two peptides in each group, the possible occurrence of two prohormones is discussed. PMID- 6277630 TI - Incorporation of N-ethylmaleimide into the membrane-bound ADP/ATP translocator. Isolation of the protein labeled with N-[3H]ethylmaleimide. AB - The incorporation of N-ethylmaleimide into the 30,000-Mr component of beef-heart mitochondria has been studied as a function of various ligands to the ADP/ATP carrier and the isolation of the N-ethylmaleimide-labeled protein is reported. 1. The incorporation of N-ethylmaleimide into the 30,000-Mr component is specifically stimulated by ADP and ATP. Thus by differential incorporation of N ethylmaleimide, the 30,000-Mr component is preferentially labeled. 2. Addition of carboxyatractylate inhibits, whereas bongkrekate tolerates, the incorporation of N-ethylmaleimide. 3. After solubilization by Triton the purification of N ethylmaleimide-labeled protein is facilitated in the presence of bongkrekate but not of carboxyatractylate, in agreement with the postulated existence of only a bongkrekate-N-ethylmaleimide-protein complex. The labeled protein was purified to homogeneity on hydroxyapatite in Triton and subsequently, after denaturation in dodecylsulfate, on Sepharose 6B. 4. The identify of the isolated labeled protein with the formerly isolated bongkrekate-protein or carboxyatractylate-protein complexes is confirmed by the isoelectric point and amino acid composition. 5. Two moles of N-ethylmaleimide must be incorporated into the 30,000-Mr component in order to inhibit fully the binding of one mole carboxyatractylate. This corresponds to one -SH group per unit. PMID- 6277631 TI - The reactivity of -SH groups in the ADP/ATP carrier isolated from beef heart mitochondria. AB - The emergence of the reactivity of -SH groups associated with conformation changes has been studied on the ADP/ATP carrier, is isolated in three different inhibitor-protein complexes. 1. The bongkrekate-protein complex incorporates approximately one molecular more of N-ethylmaleimide than the carboxyatractylate protein complex. After extensive denaturation by dodecylsulfate in urea, both inhibitor complexes exhibit four reactive -SH groups per subunit. Thus one of four -SH groups per subunit has been unmasked in the bongkrekate-protein complex. 2. The interconversion from the bongkrekate-protein complex to the carboxyatractylate-protein complex is inhibited after the -SH groups have been blocked. 3. The protein complex isolated with the more easily dissociable atractylate, is used to demonstrate, by the emergence of the -SH groups, the transition into the m-state. This transition is specifically catalyzed by ADP and ATP. 4. Using 2,2'-dinitro-5,5'-dithiodibenzoate, the appearance of the -SH groups on transition from the c-state to the m-state can be followed spectrophotometrically. The specificity for the catalyzing nucleotides is identical with that for the transport. The Km for ADP and ATP is in the range of 1 microM. In conclusion, the thiol groups of the isolated ADP/ATP carrier behave as in the mitochondrial membrane. The unmasking of -SH groups is in full accordance with the concept of two conformational states (c and m). PMID- 6277632 TI - Phospholipid interconversions in Mycoplasma capricolum. AB - Mycoplasma capricolum cells increase their phospholipid content by incorporating exogenous phospholipids from the growth medium. Growing the cells in media with increasing serum concentrations resulted in a massive incorporation of phosphatidylcholine and sphingomyelin (up to about 50% of total phospholipids) into the cell membrane. The incorporation of the exogenous phospholipids had essentially no effect on the rate of cell growth and did not decrease the overall phospholipid biosynthesis of the cells. Thus, the ratio of phospholipid to protein in membranes from cells grown with 5% horse serum was 0.5 (mumol/mg) compared to 0.3 (mumol/mg) in cells grown without serum, and the relative content of charged polar lipids was apparently decreased. The consequence of the incorporation of exogenous phosphatidylcholine was an alteration in the relative amount of the major end-products of the de novo phospholipid biosynthesis; a marked increase in the ratio of diphosphatidylglycerol to phosphatidylglycerol was observed. The possibility that the increase in the ratio of diphosphatidylglycerol to phosphatidylglycerol is part of a control mechanism to maintain a mixture of bilayer and non-bilayer lipids is discussed. PMID- 6277633 TI - Interaction of cAMP derivatives with the 'stable' cAMP-binding site in the cAMP dependent protein kinase type I. AB - cAMP binding to the 'stable' cAMP-binding sites in the regulatory subunit of the cAMP-dependent protein kinase type I was investigated using a set of 18 selected derivatives. All the tested analogues were competitive with [3H]cAMP and inhibitor constants from 12 nM to 20 microM with the free regulatory subunit were determined. The cAMP molecule seemed to be bound by these specific hydrogen bonds to the 5' and 3' oxygen, the 2' hydroxyl, and an ion pair interaction between the negative charge in equatorial position and a positively charged amino acid side chain. The adenine base is rather unspecifically bound with no hydrogen bonds involved. This binding specificity of the 'stable' site is similar to the requirement for dissociation as determined by the activation of the kinase by a respective analogue. This indicates that occupation of the 'stable' sites leads to activation of the protein kinase. The presence of the catalytic subunit reduced the affinity of most analogues. The binding of one derivative with the negative charge fixed in the axial position is not influenced by the addition of the catalytic subunit and ATP. A plausible model for a conformational change during the activation process in the 'stable' site is discussed. PMID- 6277634 TI - Studies on the transmembrane orientation of cytochrome c oxidase in phospholipid vesicles. AB - We report investigations into the direction of orientation of cytochrome c oxidase in reconstituted vesicles and the factors determining this. Measurement of the enzyme orientation employed two independent techniques: monitoring of the level of haem reduction by membrane-permeant and membrane-impermeant reagents and a kinetic analysis of the reduction of a spin label covalently bound to the oxidase surface. The method of preparation of the oxidase vesicles had a pronounced effect on the enzyme orientation and the two measurement techniques agreed in indicating that the proportion of mitochondrially oriented enzyme was approximately 85% and 50% for vesicles prepared by cholate dialysis and sonication respectively. Our results show that the membrane orientation of the oxidase is determined by interactions between the phospholipid bilayer and the portion of the enzyme embedded therein, as opposed to gross physical constraints. In particular, we demonstrate that the orientation of the oxidase is affected by the fluidity and surface charge of the membrane. PMID- 6277635 TI - Effect of thyroid hormones and cyclic AMP on some lipogenic enzymes of the fat cell. PMID- 6277636 TI - Investigation of the iron-sulfur clusters in some mitochondrial mutants of Saccharomyces cerevisiae. A possible correlation between Rieske's iron-sulfur cluster and cytochrome b. PMID- 6277637 TI - Synthesis, characterisation and biological significance of (2'-5')oligoadenylate derivatives of NAD+, ADP-ribose and adenosine(5')tetraphospho(5')adenosine. PMID- 6277638 TI - Glycolipids and opiate action. PMID- 6277639 TI - Diagnostic cytology in breast cancer screening. AB - In the period 1977-1978, 245 solid or cystic breast tumors and 134 nipple secretions were evaluated cytologically at the Trieste Cancer Institute. The results are discussed in the light of literature reports, and the high incidence of unsatisfactory smears observed in the group of malignant lesions (14,7%) is attributed to incorrect technique in specimen collection. The incidence of false positives and false negatives was 0,2% and 2,9%, respectively. The data reported are insufficient for significant statistical conclusion, but nevertheless confirm further the diagnostic importance of breast cytology whose routine practice in patient study protocols for breast cancer is highly recommended. PMID- 6277640 TI - Nephroblastoma associated with mesangioproliferative glomerulonephritis. AB - The case of a girl showing left-sided Wilms tumor together with a nephrotic syndrome is presented. Recovery after tumor nephrectomy was uneventful. After cessation of cytostatic chemotherapy chronic renal failure developed. Open wedge biopsy of the right kidney revealed endstage glomerular disease. Renal histology in patients with this rare combination and in those presenting with male pseudohermaphroditism and nephron disorders shows a specific pattern. It is suggested that there is a genuine type of nephropathy linked either to Wilms' tumor and/or to male pseudohermaphroditism. PMID- 6277641 TI - Specific triiodothyronine binding by tumor cells and spleen cells in a thyroid hormone dependent mouse tumor system. PMID- 6277642 TI - Failure of TRH and ORG 2766 hexapeptide to counteract alcoholic inebriation in man. AB - The actions and interactions of alcohol (1.5 g/kg) and two stimulant peptides were investigated in healthy volunteers. Double blind and cross over studies in 8 subjects, at one-week intervals, revealed that ORG 2766 (5 or 20 mg i.m.) and particularly TRH (10 micrograms/kg i.v.) enhanced rather than antagonized alcohol induced inebriation. The interactions were associated with elevated breath alcohol concentrations. Similar studies with 8 other subjects showed that peptides given in combination with a non-alcoholic drink tended to impair performance assessed as various psychophysiological variables. However, ORG 2766 5 mg was subjectively rated as producing a feeling of improved performance. PMID- 6277643 TI - Epstein-Barr virus (EBV) receptor implantation onto human B lymphocytes changes immunoglobulin secretion patterns induced by EBV infection. AB - Mosaic membrane vesicles containing both Epstein-Barr virus (EBV) receptors and Sendai virus envelope proteins were allowed to form by the previously described membrane solubilization and co-reconstitution technique. The vesicles were allowed to fuse with the membranes of normal human B lymphocytes, whereafter the cells were infected with transforming EBV (B95-8 substrain). Compared to similarly infected but otherwise unmanipulated cells, the receptor-implanted lymphocytes responded with a larger number of EBV-determined nuclear antigen positive and immunoglobulin-secreting plaque-forming cells (PFC). Moreover, there was a clear increase of the IgG/IgM PFC ratio in the receptor-implanted B lymphocytes. These results show that not all human B lymphocytes that can potentially be activated by EBV express functional EBV receptors. B lymphocytes programmed to secrete IgG appear to be more defective in this respect than IgM secretors. PMID- 6277644 TI - Possible regulatory role of dynorphin-(1-13) on narcotic-induced changes in naloxone efficacy. AB - Pretreatment of mice with a single injection of morphine, beta-endorphin, Leu enkephalin, FK33824 or [D-Ala2-D-Leu5]enkephalin, for 3 h increased the ability of naloxone to antagonize the analgesic effects of morphine. However, dynorphin (1-13) can only antagonize morphine, beta-endorphin or Leu-enkephalin induced increased naloxone efficacy but not FK33824 or [D-Ala2-D-Leu2]enkephalin. Dynorphin itself can also increase naloxone efficacy when treated alone. However, this effect can be prevented when animals are pretreated with dynorphin and naloxone together. PMID- 6277645 TI - Modification of diazepam's antileptazol activity by endogenous tryptophan-like compounds. AB - Three possible endogenous ligands for benzodiazepine receptors, beta-carboline-3 carboxylic acid ethyl ester (beta CCE), tryptophylglycine (Trp-Gly) and ACTH peptides, have been tested, following intracerebroventricular (i.c.v.) administration in mice, for their direct effects on the CNS and the modification of diazepam's antileptazol activity. Whilst beta CCE and ACTH both reduced diazepam's antileptazol activity, only beta CCE had direct stimulant effects. Trp Gly produced sedation and augmented the antileptazol effect of diazepam. Despite these differing effects, it remains possible that the endogenous ligand for the benzodiazepine receptors has a tryptophan-like structure. PMID- 6277646 TI - Interactions of beta-carbolines with the benzodiazepine receptor: structure activity relationships. AB - The effects of a number of beta-carboline derivatives on [3H]flunitrazepam binding to the benzodiazepine binding site were investigated. The potency of beta carbolines at the benzodiazepine binding site appeared to be determined largely by the aromaticity of the molecule. Norharmane-3-carboxylic acid ethyl ester was considerably more potent (IC50 10 nM) than its tetrahydro-beta-carboline analogue (IC50 6 microM). There is essentially no difference in potency between the (+)- and (--)-forms of the tetrahydro-beta-carboline-3-carboxylate analogues. PMID- 6277647 TI - Evidence against adenylate cyclase-coupled adenosine receptors in the guinea pig heart. AB - (--)-N6-(Phenylisopropyl)adenosine (PIA, 10 microM) and 5'-N (ethylcarboxamide)adenosine (NECA, 50 microM), potent agonists at the adenylate cyclase-coupled R-site adenosine receptor, were investigated for adenylate cyclase inhibition in a guinea pig ventricular membrane preparation by means of the dATP assay method. Neither compound influenced basal or isoproterenol stimulated cyclase activity, irrespective of whether Na ions were present or not. These data suggest that R-site receptors may not be involved in the cardiac adenylate cyclase system of the guinea pig. PMID- 6277648 TI - Discrepancy in the regulatory effects of sodium and guanine nucleotides on adrenaline and clonidine binding to alpha 2-adrenoceptors in human fat cell membranes. AB - Guanine nucleotides and sodium were found to reduce the affinity of adrenaline for the alpha 2-adrenoceptor sites labelled by [3H]yohimbine in human fat cell membranes. Surprisingly, the negative regulation by these agents was far weaker when clonidine, the alpha 2-agonist prototype was used. The results are discussed in relation to the weaker or the lack of adenylate cyclase inhibition promoted by clonidine and indicate that the formation of the lower affinity state of the alpha-receptor is essential to the alpha 2-adrenoceptor-mediated adenylate cyclase inhibition. PMID- 6277649 TI - ACTH1-24 increases stimulation-evoked noradrenaline release from sympathetic nerves by acting on presynaptic ACTH receptors. PMID- 6277650 TI - Evidence for the presence of P1-purinoceptors on cholinergic nerve terminals in the guinea-pig ileum. AB - The inhibitory effects of adenosine, ATP, 5'-adenylyl methylene diphosphonate (beta, gamma-meATP) and adenosine 5'-alpha, beta-methylene triphosphonate (alpha, beta-meATP) were compared on the cholinergic twitch responses to transmural stimulation of the guinea-pig ileum. Adenosine, ATP and beta, gamma-meATP reduced the twitch responses in a concentration dependent manner. Theophylline antagonized and dipyridamole potentiated the inhibitory responses to adenosine, ATP and beta, gamma-meATP. Inhibitory responses to alpha, beta-meATP were usually preceded by an enhancement in twitch height. Contractions of the unstimulated ileum to alpha, beta-meATP were blocked by atropine and tetrodotoxin while those elicited by ATP were unaffected, which suggests that the initial excitatory effects of alpha, beta-meATP may be due to its ability to release ACh from cholinergic nerve terminals. Use of high pressure liquid chromatography and bioluminescence assay techniques demonstrated the ability of the tissue to degrade ATP and beta, gamma-meATP and, at a much slower rate, alpha, beta-meATP. Inhibitory responses to ATP, AMP and beta, gamma-meATP were reduced by adenosine deaminase, which also abolished responses to adenosine. 5'-AMP deaminase abolished responses to AMP and adenosine, and reduced those to ATP and beta, gamma-meATP. The results suggest that the inhibitory effect of ATP on cholinergic neurotransmission is due to its rapid breakdown to AMP or adenosine, which act on prejunctional P1-purinoceptors. PMID- 6277651 TI - Effect of Nin-formylation of the tryptophan residue on gastrin (HG-13) binding and on gastric acid secretion. AB - The biologically active iodinated [Leu11]human gastrin-13 was found to exhibit high affinity for gastric mucosal cell receptors. We compared its biological activity on rat stomach and its receptor affinity on rat gastric mucosal cells isolated with pronase with those of a reversibly modified derivative obtained by direct Nin-formulation of the Trp10 residue. The results strongly suggested the involvement of the indole-NH group in both the specific binding of gastrin and the triggering of acid secretion. PMID- 6277652 TI - Loss of rat cerebral cortical opiate receptors following chronic desimipramine treatment. AB - The characteristics of central opiate and beta-adrenergic receptors were studied after giving the tricyclic antidepressant desimipramine (DMI) to rats. Administration of DMI for 1 or 7 days did not alter cortical opiate or beta adrenergic receptors. However, after 21 days of treatment there was a significant subsensitivity of both these receptors in the cortex. A specific loss of receptor sites for opiate binding was detected with no change in affinity. After a similar time of treatment, opiate receptors in the corpus striatum and hippocampus were the same as the controls. The data suggest that change in cortical opiate receptors may be important in the mechanism of action of antidepressants. PMID- 6277654 TI - Difference in tolerance development of hypothermia and pentobarbital-induced sleep prolongating effect of 11-hydroxy-delta 8-tetrahydrocannabinol and 11-oxo delta 8-tetrahydrocannabinol in mice. AB - Daily administration (5 mg/kg i.v.) of 11-hydroxy-delta 8-tetrahydrocannabinol or 11-oxo-delta 8-tetrahydrocannabinol as well as delta 8-tetrahydrocannabinol quickly induced tolerance to their hypothermic effects in mice. Tolerance also developed to their pentobarbital-induced sleep prolongating effects. However, the degrees of tolerance development differed from one another. The sleeping times after the 8th injection of these cannabinoids were still significantly longer than those of the controls when no hypothermia was induced. PMID- 6277653 TI - Depressor effects of isoguvacine propyl ester and isoarecaidine propyl ester due to stimulation of central GABA receptors in the cat. AB - Upon their simultaneous administration into the left and right vertebral artery of the cat, 2 x 30 micrograms isoarecaidine propyl ester (IAPE) and 2 x 30 micrograms isoguvacine propyl ester (IGPE) induce hypotension and bradycardia. These effects were antagonized by (+)-bicuculline or picrotoxin. IAPE is rapidly hydrolyzed by cat brain tissue in vivo. It is suggested that both IAPE and IGPE are prodrugs and that the corresponding carboxylic acids are GABA agonists and responsible for the cardiovascular effects evoked. PMID- 6277655 TI - The effect of GTP on benzodiazepine receptors. AB - GTP decreases the specific binding of [3H]flunitrazepam to cerebral cortical membrane sites. Scatchard analysis data show that GTP does not alter the maximum binding sites (Bmax), but increases the dissociation constant (KD) for [3H]flunitrazepam. GTP also inhibits the specific binding of the benzodiazepine antagonist [3H]CGS-8216. The specific binding of [3H]CGS-8216 displaceable by clonazepam is inhibited to a greater extent by GTP than that displaceable by the benzodiazepine antagonists, CGS-8216 or beta-CCE. PMID- 6277656 TI - The increase of cardiac beta 1-subtype of beta-adrenergic receptors in adult rats following neonatal 6-hydroxydopa treatment. AB - An analysis of Hofstee plots of specific [3H]dihydroalprenolol ([3H]DHA) binding in the presence of various concentrations of practolol showed that KD and Bmax in beta 1-subtype adrenoceptors were 19.2-fold lower and 2.76-fold higher than those in beta 2-subtype in adult rat hearts, respectively. Neonatal treatment with 6 hydroxydopa produced a significant increase only in Bmax in beta 1-adrenoceptors without changing Bmax and KD in beta 2-receptors. "Up' regulation in the density of beta 2-type of adrenoceptors seems to be caused by the 6-hydroxydopa-induced sympathetic lesion. PMID- 6277657 TI - Dopaminergic inhibition of cardiac sympathetic nerve function by pergolide. AB - The effect of pergolide, a dopaminergic ergoline, on cardiac sympathetic neurotransmission was examined in pentobarbital-anesthetized dogs. Positive chronotropic responses to right postganglionic cardioaccelerator nerve stimulation were significantly reduced following infusion of pergolide, 1.25 microgram/kg per min for 10 min, whereas positive chronotropic responses to exogenous norepinephrine were unchanged. Pretreatment with sulpiride but not with yohimbine completely antagonized the inhibitory effect of pergolide. These results demonstrate that pergolide inhibits sympathetic neurotransmission to the heart by a specific activation of presynaptic dopamine receptors. PMID- 6277659 TI - Evidence that dynorphin-(1-13) acts as an agonist on opioid kappa-receptors. AB - The study concerned the opioid-receptor subtype on which dynorphin-(1-13) acts in in vitro isolated preparations. The potency of dynorphin-(1-13) relative to that of ethylketocyclazocine (Mr 2266), a representative kappa-receptor agonist, in inhibiting the electrically evoked contractions of the guinea-pig ileum was found to be similar to that found with either mouse was deferens or rabbit ileum. Moreover, Mr 2266 was found to be several-fold more effective than naloxone to antagonize the agonist actions of both kappa-receptor agonists such as ethylketocyclazocine, ketocyclazocine and bremazocine, and dynorphin-(1-13) either in the guinea-pig ileum, mouse vas deferens, or in rabbit ileum. Additionally, dynorphin-(1-13) was found to have a significant inhibitory action on the rabbit vas deferens which had been shown to contain kappa-receptors exclusively. The data indicate that dynorphin-(1-13) acts as an endogenous agonist on kappa-receptors. PMID- 6277658 TI - An orally effective, long-acting dopaminergic prodrug: (-)-10,11-methylenedioxy-N propylnoraporphine. PMID- 6277660 TI - ACTH1-24 counteracts the prolactin-releasing effect of an opioid. AB - The administration of the synthetic and stable opioid peptide[D Ala2,MePhe4,Met(O)5o1]enkephalin (FK 33-824) at the dose of 0.2 mg/kg i.v. induced a rise in plasma levels of prolactin in the rat, an effect which was prevented by 1 mg/kg i.v. of the opiate antagonist naloxone. A simultaneous i.v. injection of 20 microgram/kg i.v. of ACTH1-24 significantly reduced the prolactin releasing effect of FK 33-824, therefore giving further evidence of an in vivo interaction between ACTH-related peptides and opioid peptides. PMID- 6277661 TI - Anticonvulsant effects of clonidine mediated through central alpha2 adrenoceptors. AB - The effects of centrally (0.3-30 ng/kg) and peripherally (0.01 microgram/kg-1.0 mg/kg) administered clonidine on PTZ-induced seizures was studied in rats. At low doses, dose-dependent decreases in the duration of seizures was observed but at higher doses the duration returned to control levels. Anticonvulsant activity was antagonized by yohimbine (100 microgram/kg i.p.) indicating alpha2-adrenoceptor involvement, whereas the second phase of the response was antagonized by prazosin (10 ng/kg i.c.v.) indicating that it involved alpha1-adrenoceptors. PMID- 6277662 TI - Presynaptic dual inhibitory actions of guanabenz on adrenergic transmission. AB - Actions of guanabenz, clonidine and guanethidine on adrenergic transmission were studied in rabbit atria and ilea with intact nerves. While guanabenz 3 X 10(-6) M and clonidine 3 X 10(-7) M inhibited to a similar degree atrial positive chronotropic responses to nerve stimulation at 2 Hz, no inhibition was seen at 10 Hz. Phentolamine 3 X 10(-6) M completely antagonized the inhibition by clonidine, but a partial antagonism was seen against guanabenz. Cocaine 10(-5) M prevented the phentolamine-resistant inhibition by guanabenz. Guanethidine 10(-5) M inhibited the responses to 2 and 10 Hz. In ilea, guanabenz 10(-5) M inhibited frequency-independent relaxation responses to 2-20 Hz. Clonidine 10(-5) M inhibited preferentially the responses to lower frequencies. After the temperature of the bathing solution was kept at 4 degree C during the period of drug application, guanabenz no longer produced an inhibition of the transmission, whereas clonidine produced the usual inhibition. Thus, guanabenz appears to have both presynaptic alpha-receptor stimulating and adrenergic neuron blocking actions. PMID- 6277663 TI - Age-related decrease of in vitro isoproterenol-induced cyclic AMP accumulation in rat aorta. AB - Isoproterenol-induced accumulation of cyclic AMP was investigated in aorta rings from 7 to 18 weeks old rats. The effect of isoproterenol on cyclic AMP reached a maximum in 1 min. The concentration-effect curves showed a significant decrease of the maximal accumulation of cyclic AMP as a function of age, without any variation in the effective concentration range. These results parallel the previously reported decrease of beta-adrenergic relaxation of rat aorta with age. PMID- 6277664 TI - Temperature dependence of [3H]dihydroalprenolol binding in rat myocardium. AB - The beta-adrenergic antagonist, [3H]dihydroalprenolol, was used to label binding sites in crude rat myocardial plasma membranes. The specificity of binding was dependent on the temperature of the assay. Specific binding at 22 and 37 degree C and at concentrations of radioligand less than 5 nM was consistent with binding to the myocardial beta-receptor. Binding sites labeled at 4 degree C possessed quite different properties. Binding was non-stereoselective and of lower affinity. Agonist compounds were much less effective at competing for the labeled myocardial sites at 4 degree C than at 22 degree C. Those beta-antagonists which additionally possess pharmacological "quinidine-like' activity (e.g. propranolol, alprenolol) were potent competitors at 4 degree C, but competition was non stereoselective. In contrast atenolol, a beta-antagonist devoid of "quinidine like' activity was ineffective at 4 degree C. Furthermore, procaine, and quinidine itself were potent competitors of [3H]dihydroalprenolol binding at 4 degree C. Thus the specificity of [3H]dihydroalprenolol binding to rat heart membranes at 4 degree C appears to be directed non-stereoselectively at that portion of the competing molecule which recognized "quinidine-like' as opposed to adrenergic activity. PMID- 6277665 TI - Haemophilus influenzae induced loss of lung beta-adrenoceptor binding sites and modulation by changes in peripheral catecholaminergic input. AB - Vaccination of guinea pigs with killed suspensions of Haemophilus influenzae, a bacterium often found in the deeper respiratory airways of asthmatic bronchitics, results in a number of effects suggesting an impairment of beta-adrenoceptor function. A [3H]dihydroalprenolol binding assay was used to determine the number of beta-adrenoceptors (Bmax) following H. influenzae vaccination. The Bmax declined significantly by 29% from 1240 +/- 80 to 880 +/- 70 fmol/mg protein, while the binding affinity of the sites was not changed. Specific binding in the presence of 1.8 nM [3H]DHA to tracheal longitudinal smooth muscle was also significantly lower in H. influenzae-vaccinated animals as compared to controls. Furthermore modulation of peripheral sympathetic input to lung beta-adrenoceptors was evaluated in our model. Pretreatment with Ro4-4602, an inhibitor of dopa decarboxylase, increased the number of beta-adrenoceptors and prevented the H. influenzae-induced loss of beta-adrenoceptors. On the other hand repeated doses of the antidepressant desipramine mimicked the effect of H. influenzae vaccination i.e. a loss of beta-adrenoceptors. Desipramine and H. influenzae vaccination were not synergistic in their effects. The effects of H. influenzae and modulation of catecholaminergic input on guinea pig lung beta-adrenoceptors were compared with tracheal strip relaxation by isoproterenol, following similar treatments assessed in a superfusion model. Changes in lung beta-adrenoceptor number were almost identical with changes in tracheal strip relaxation. These results suggest that compensatory regulation adapts the number of respiratory beta-adrenoceptors to changes in sympathetic input. A similar mechanism may underlie the loss of beta-adrenoceptors following H. influenzae vaccination. PMID- 6277667 TI - Local anaesthetic activity of organic calcium antagonists: relevance to their actions on smooth muscle. AB - Local anaesthetic activity was measured on the rat phrenic nerve at 37 degrees C: Verapamil HCl had 5.1X and methoxyverapamil HCl had 3.0X the potency of lidocaine. Flunarizine was about equipotent with lidocaine, while nifedipine, diazoxide and sodium nitroprusside were without effect. It is concluded tht the smooth muscle inhibitory actions of the calcium antagonists, even when used in concentrations possessing local anaesthetic activity, are due to a reduction in Ca2+ permeability. In rat isolated vasa deferentia, lidocaine had both stimulant and inhibitory actions. On its own it induced rhythmic contractions (not blocked by phentolamine) and it also augmented the frequency of rhythmic contractions produced by methoxamine or barium. These actions may be due to block of K+ channels. Lidocaine also reduced the amplitude of methoxamine and barium induced contractions and both phases of the KCl contraction. The KCl response was restored by increasing "Ca2+]0, and it is therefore concluded that lidocaine was acting by blockade of Ca2+ channels. PMID- 6277666 TI - Bronchoselective actions of a new series of trimetoquinol analogues. AB - In selected beta 1- (guinea pig atrial) and beta 2- (guinea pig trachea and lung parenchyma) adrenoceptor systems, we have examined the interaction of isoproterenol (ISO), trimetoquinol (TMQ), erythro- and threo-diastereoisomers of 1-(3,4,5-trimethoxy-alpha-methylbenzyl)-6,7-dihydroxy-1,2,3,4 tetrahydroisoquinoline (alpha-methylTMQ), alpha-dimethylTMQ, N-methylTMQ and N-[2 methyl-2-(3,4,5-trimethoxyphenyl)propyl]dopamine (open chain dimethylTMQ analogue). The rank order of potency for agonists in trachea was threo-alpha methylTMQ greater than (+/-)-TMQ greater than ISO greater than erythro-alpha methylTMQ greater than N-methylTMQ greater than alpha-dimethylTMQ. Only N methylTMQ gave an intrinsic activity similar to ISO, whereas the alpha-methylated TMQ analogues were partial agonists in this beta 2-system. In atria, the rank order of beta 1-potency was ISO greater than (+/-)-TMQ greater than threo-alpha methylTMQ greater than N-methylTMQ = erythro-alpha-methylTMQ. Maximal chronotropic effects of all compounds, with the exception of threo-alpha methylTMQ, were similar to ISO in this preparation. Both alpha-dimethylTMQ and open chain dimethylTMQ analogues were inactive as agonists in this beta 1-system. The ratio of beta 2 : beta 1 selectivity (trachea vs. atria), relative to ISO for threo-alpha-methylTMQ, erythro-alpha-methylTMQ, TMQ and N-methylTMQ was 106.5, 27, 7 and 5.8, respectively. Whereas the rank order of potency for selected compounds in lung parenchyma was ISO greater than threo-alpha-methylTMQ = TMQ greater than erythro-alpha-methylTMQ, the comparative beta 2-selectivity (lung parenchyma vs. atria) relative to ISO, for erythro-alpha-methylTMQ, threo-alpha methylTMQ and TMQ was 2.5, 1.9 and 0.24, respectively. It is concluded that lipophilic substitutions on the alpha-carbon of the 1-(3,4,5-trimethoxybenzyl) substituent of TMQ can generate compounds which are potent bronchoselective adrenoceptor agonists. Threo-alpha-methylTMQ and erythro-alpha-methylTMQ were more beta 2-selective than (+/-)-TMQ. PMID- 6277668 TI - Effect of aldosterone antagonists on mineralocorticoid synthesis in vitro. Inhibition of aldosterone production by prorenoate-K. AB - A perifusion technique using frog adrenal glands has been applied to investigate the effects of long-term administration of a new aldosterone antagonist (potassium prorenoate; SC 23992) on mineralocorticoid production. Whatever the duration of administration of potassium prorenoate, at a constant concentration of 5 X 10(-4) M, a significant inhibition of aldosterone output occurred during the passage of the compound. The inhibition was immediate (lag period less than 10 min); the amplitude of the inhibition was constant during the whole experiment and ranged from 77 to 89%; the aldosterone output returned to a regular basal value 80-100 min after the end of infusion of potassium prorenoate. We have also investigated the effect of a concentration gradient of potassium prorenoate (similar to the concentration gradient of aldosterone antagonist observed in plasma after a single oral administration of the molecule) upon aldosterone production over 12 h. From this study, we have established the existence of a highly significant correlation between the extent of the inhibition of aldosterone production and the concentration of the aldosterone antagonist. Finally we have observed that potassium prorenoate blocked the stimulation of aldosterone secretion induced by synthetic ACTH and significantly reduced the angiotensin-induced aldosterone stimulation. The present results indicate that, besides the well-known competitive inhibition of aldosterone binding exerted by potassium prorenoate at the renal receptor site, a direct inhibition of aldosterone biosynthesis also accounts for the pharmacological activity of this aldosterone antagonist. PMID- 6277669 TI - Mechanism of cadmium-induced blockade of neuromuscular transmission. PMID- 6277670 TI - Drug affinities for the agonist and antagonist states of the opioid receptor. AB - Traditional methods of analyzing the binding of opiates do not accurately establish the affinities of the drugs for the agonist and antagonist states of the opioid receptor. The poor results obtained by the Scatchard procedure may be attributed to the presence of extraneous drug binding sites in the brain homogenate medium. In this study a nonlinear regression technique has been employed to analyze receptor binding in terms of the two-state Snyder-Pert model while simultaneously considering drug association with the aggregate of non receptor binding sites. Results are reported for naloxone, pentazocine, and dihydromorphine, which respectively represent a pure antagonist, a mixed agonist antagonist, and a pure agonist. the affinity constants characterizing equilibrium binding at the agonist and antagonist states of the receptor at 37 degrees C are given for each of the three drugs. In addition, a theoretical index, f tau, is defined which provides a measure of the fraction of receptors existing in the agonist state. It is found that the graft of f tau vs log [D] exhibits features similar to the in vivo dose-response curves for the drugs. PMID- 6277671 TI - A comparison of inotropic actions of p-chloromercuribenzoate, ouabain, digitoxin and digitoxigenin. PMID- 6277673 TI - Interaction of avermectins with [3H]beta-carboline-3-carboxylate ethyl ester and [3H]diazepam binding sites in rat brain cortical membranes. AB - The binding of [3H] beta-carboline-3-carboxylate ethyl ester ([3H] beta-CCE), a ligand for the benzodiazepine receptor in the mammalian CNS, to rat cortical membranes, is enhanced by avermectin B1a and its therapeutic formulation, Ivermectin. In contrast to the effects of the avermectins on [3H]diazepam binding, which involve changes in both receptor affinity and number, increases in beta-CCE binding, which are much less than those observed for the benzodiazepine ligand, involve only alterations in receptor number. This Bmax increase is bicuculline insensitive whereas Ivermectin effects on benzodiazepine binding are partially antagonized by GABA antagonist. The data suggest a differential interaction by the avermectins on benzodiazepine and beta-CCE binding sites in rat cortical membranes and indicate that these macrolide anthelmintics may be a useful tool for characterizing benzodiazepine/anxiolytic receptor subtypes. PMID- 6277672 TI - Vasopressin and beta-endorphin release after osmotic and non-osmotic stimuli: effect of naloxone and dexamethasone. AB - The purpose of this study was to determine whether or not vasopressin release in response to various stimuli in the conscious rat is controlled by endogenous opioid peptides, in particular beta-endorphin. Naloxone (1 mg.kg-1 i.m.) promoted vasopressin release in response to both an angiotensin II infusion (500 ng . kg-1 . min-1) or an isosmolar, nonhypotensive hypovolaemia achieved by polyethylene glycol injection (PEG, 20% solution i.p.); however, naloxone was without effect when vasopressin release was induced by hypertonic saline injection (2.5% solution i.p.) or a severe fall in arterial blood pressure following trimethidinium (10 mg . kg-1 i.m.) induced ganglionic blockade. Vasopressin release was accompanied by an increase in plasma beta-endorphin-like immunoreactivity (beta-EI) following an angiotensin II infusion of PEG administration, but not after hypertonic saline or trimethidinium injection. Dexamethasone pretreatment (0.5 mg . kg-1 twice i.p.) prevented the increase in plasma beta-EI following an angiotensin II infusion or PEG administration. The simultaneous angiotensin II- or PEG-induced increase in vasopressin release was unaffected or potentiated, respectively, by the glucocorticoid. In contrast, vasopressin release in response to hypertonic saline or trimethidinium injection was significantly inhibited by dexamethasone. We conclude that an inhibitory control by endogenous opiates is involved in some, but not all of the different pathways leading to vasopressin release. The results obtained do not prove but can be reconciled with the proposal that hypophyseal beta-endorphin is the compound responsible. PMID- 6277674 TI - Interaction of the putative dopamine autoreceptor agonists, 3-PPP and TL-99, with the dopamine-sensitive adenylate cyclase of carp retina. AB - The putative dopamine autoreceptor agonists, 3-PPP and TL-99 were examined for their ability to stimulate postsynaptic dopamine receptors associated with adenylate cyclase (D1-receptors) in the carp retina. In intact pieces of retina, 3-PPP had no significant effect on cyclic AMP production at concentrations up to 100 microM, whereas the aminotetralin, TL-99, caused a concentration-dependent increase in cyclic AMP levels with an approximate EC50 of 3.6 microM. Dopamine and ADTN had EC50 values of 3.5 and 3.1 microM respectively. Furthermore, in homogenates of the retina, 100 microM ADTN and 100 microM TL-99 stimulated adenylate cyclase activity 92 and 79% respectively as compared to the stimulation evoked by 100 microM DA. In contrast, 100 microM 3-PPP was essentially inactive at stimulating adenylate cyclase in carp retinal homogenates. These findings suggest that TL-99 can interact with postsynaptic D1-receptors and is not as selective a dopamine autoreceptor agonist as 3-PPP, which has no apparent activity at the D1-receptor. PMID- 6277675 TI - Sex and strain differences in benzodiazepine receptor binding in Roman rat strains. AB - Specific [3H]flunitrazepam binding was determined for five brain regions in both sexes of the three Roman rat strains. Small strain differences and a large Sex Region interaction were detected, benzodiazepine receptor binding being significantly higher in females in the two cortical areas and the cerebellum, but significantly lower in the striatum and hippocampus. These findings are discussed in the context of sex and strain differences in behavior and benzodiazepine sensitivity. PMID- 6277676 TI - Effect of yohimbine and its diastereoisomers on clonidine-induced depression of exploration in the rat. AB - The effects of yohimbine, rauwolscine and corynanthine on clonidine-induced depression of exploration (ambulation and rearing) in the rat were investigated. Yohimbine and rauwolscine antagonized clonidine-induced hypoactivity in a dose range of 0.3 - 3 mg/kg i.p. By contrast, corynanthine was found to lack such an effect up to ten times higher doses. These results are in agreement with the reported differential affinity of these drugs for the alpha 2-adrenoceptors respectively and suggest the involvement of alpha 2-receptors in the mediation of depressant effect of clonidine. The antagonism of clonidine-induced hypoactivity appears, therefore, to provide a suitable in vivo measure of preferential alpha 2 adrenoceptor blocking activity of drugs. PMID- 6277677 TI - Interaction of p-nitrophenylalanine enkephalins with mu-, delta- and kappa subtypes of the opiate receptor. AB - Substitution of L-p-nitrophenylalanine for phenylalanine in position 4 of [D Ala2, KLeu-NH2(5)]enkephalin analogues increased their affinity for mu-, delta- and kappa-binding sites in guinea-pig brain, increased their potency to inhibit electrically evoked contractions of the guinea-pig ileum and mouse vas deferens, and increased their analgesic potency in the mouse tail flick test, Introduction of L-adamantylalanine, but not of L-neopentylglycine, in position 5 resulted in a loss of activity. PMID- 6277678 TI - [3H]CL 218,872, a novel triazolopyridazine which labels the benzodiazepine receptors in rat brain. AB - We examined the specific binding of [3H]CL 218,872, a novel triazolopyridazine to benzodiazepine receptors in the rat cerebral cortex. Two binding sites with KD values of about 10-30 nM and 200-600 nM were demonstrated. A number of benzodiazepine anxiolytics inhibited [3H]CL 218,872 binding in a manner which correlates with the potency of these drugs for inhibiting [3H]benzodiazepine binding. Our initial studies show that [3H]CL 218,872 can label the benzodiazepine receptors and may prove to be a useful probe for studying benzodiazepine heterogeneity and regulation. PMID- 6277679 TI - Stimulation of GABA receptor binding by barbiturates. AB - Barbiturates increase Na+-independent GABA binding to bovine cerebral cortex membranes. The relative activity of a series of compounds to enhance GABA binding correlates significantly with their anesthetic activity and with their ability to reverse bicuculline antagonism of GABA responses. There are regional differences in the maximal percent stimulation of GABA binding by pentobarbital; a low stimulation in cerebellum. PMID- 6277680 TI - Mixed type inhibition of [D-Ala2, D-Leu5]enkephalin binding to mu-opiate binding sites by mu-, but not by kappa-opiate ligands. AB - The interaction of the delta-opiate agonist [D-Ala2,D]Leu5]enkephalin (DADL) with the mu-opiate ligands dihydromorphine (DHM) and naloxone and the kappa-opiate ligand ethylketocyclazocine (EK) was studied in rat diencephalic membranes which contain mainly mu-opiate binding sites. Saturation binding experiments performed with tritiated DADL in the presence of DHM and naloxone were indicative of mixed type inhibition whereas EK behaved as a competitive inhibitor. This result indicates differences in the interaction of mu-, delta- and kappa-ligands with mu opiate binding sites. PMID- 6277681 TI - Inhibition of adenylate cyclase activity in the corneal epithelium by anti inflammatory steroids. PMID- 6277682 TI - Degradation and resynthesis of cyclic 3', 5' -guanosine monophosphate in truncated rod photoreceptors from bovine retina. PMID- 6277683 TI - Comparison of colony stimulating activities secreted into mouse lung conditioned medium in the presence and absence of lithium chloride. AB - Inclusion of lithium chloride during the process of secretion of granulocyte macrophage colony stimulating activity (CSA) into mouse lung conditioned medium resulted in an increased production of CSA. CSA produced in the presence of lithium chloride was compared to the CSA produced in its absence by means of ammonium sulfate precipitation, Sephadex G-1CO (S) chromatography, concanavalin A Sepharose chromatography and polyacrylamide gel electrophoresis. It was found that the two activities behaved identically, indicating that lithium-induced increase in CSA does not involve a qualitative change in the molecule of the colony stimulating factor, but reflects an increased production (or release) of a CSF molecule that is similar to the one produced in its absence. PMID- 6277684 TI - Stimulation of sporulation of Clostridium perfringens by papaverine. AB - Papaverine induced sporulation in Clostridium perfringens, strains FD-1 and PS52; growth was markedly slowed under these conditions. Papaverine induced sporulation in the presence of glucose, a sporulation repressor, although increasing glucose concentrations overcame the papaverine effect. Papaverine induced sporulation of strain FD-1 more effectively than did theophylline. PMID- 6277685 TI - Binding of 3H-dihydroalprenolol to beta-adrenoceptors in adipocytes of spontaneously hypertensive rats and essentially hypertensive patients. PMID- 6277686 TI - Effect of low levels of nitrogen dioxide inhalation on endogenous retrovirus gene expression. PMID- 6277687 TI - [Opioid peptides]. PMID- 6277688 TI - Existence of several homologous sequences in the Escherichia coli chromosome to the gene for the major outer membrane lipoprotein. PMID- 6277689 TI - A new restriction endonuclease BcnI from Bacillus centrosporus RFL 1. PMID- 6277690 TI - Characterization of the adenine nucleotide translocase of pancreatic islet mitochondria. PMID- 6277691 TI - A molecular basis for the interactions of corticotropin with opiate receptors. PMID- 6277692 TI - Characterization of [3H]NoHOQnO binding to purified complex III. PMID- 6277693 TI - The inhibition of vitamin K-dependent carboxylase by cyanide. PMID- 6277694 TI - Copper-dependent hydroxyl radical damage to ascorbic acid: formation of a thiobarbituric acid-reactive product. PMID- 6277695 TI - [Congenital defects of hand development]. PMID- 6277696 TI - Teratology study of intravaginally administered nonoxynol-9-containing contraceptive cream in rats. AB - Pregnant Long-Evans Hooded rats were intravaginally administered nonoxynol-9, a non-ionic surfactant used as a spermicidal agent in Delfen Contraceptive Cream (Ortho Pharmaceutical Corporation, Raritan, N.J.) at dosages of 4 or 40 mg/kg/day (2 and 20 times the clinical usage) on days 6 through 15 of gestation. Untreated, vehicle and sham control groups were also incorporated into the study protocol. No meaningful differences were observed between the treated and control groups in maternal toxicity, gross and microscopic appearance of vagina, maternal reproductive parameters, fetal toxicity, and the incidence of visceral and skeletal malformations. PMID- 6277697 TI - Current status of estrogen therapy for the menopause. PMID- 6277698 TI - Endometrial hyperplasia in women on cyclic or continuous estrogen regimens. AB - Twenty-five symptomatic postmenopausal women with an intact uterus were assigned in random double-blind fashion to receive 0.625 mg of conjugated estrogens on either a cyclic (3 weeks on, 1 week off) or continuous (daily) basis. The incidence of endometrial hyperplasia as demonstrated by screening biopsies at 6 and 12 months of therapy was 4.5 per 100 woman-months in the cyclic group and 3.7 per 100 woman-months in the continuous group (a difference not statistically significant). Thus, in this study, cyclic therapy was found to offer no advantage over continuous therapy. In our opinion, the rate of hyperplasia development in both groups unacceptably high, and efforts must be directed at reducing its incidence primarily. PMID- 6277699 TI - The prognostic value of acute adrenal suppression and stimulation tests in hyperandrogenic women. AB - A group of 106 consecutively seen hyperandrogenic women were subjected to an acute adrenal suppression and stimulation test. The results of these tests were analyzed with respect to androgen suppression achieved after chronic glucocorticoid therapy in the same patients. The data suggested that an acute dexamethasone test may identify the group of hyperandrogenic women who respond poorly to chronic glucocorticoid therapy. This group of patients were found to have elevated luteinizing hormone (LH) levels and LH/follicle-stimulating hormone (FSH) ratios, suggesting the possibility of an LH-related hyperandrogenism. In patients whose elevated testosterone levels were suppressed by dexamethasone, adrenocorticotropic hormone (ACTH) induced a prompt return of the testosterone levels to baseline, suggesting an ACTH-dependent hyperandrogenism. In these patients, the degree of testosterone suppression after dexamethasone was not quantitatively related to the degree of testosterone suppression after chronic glucocorticoid therapy. In all cases chronic therapy resulted in a greater suppression of androgen levels than the acute dexamethasone test. In conclusion, an acute dexamethasone suppression test appears to be of clinical value in the management of the hyperandrogenic female, particularly in identification of women who will not respond to chronic glucocorticoid suppression therapy. PMID- 6277700 TI - Liver function tests in women using intravaginal spermicide nonoxynol-9. PMID- 6277701 TI - THC effects on fructose utilization and sperm motility. PMID- 6277702 TI - [Dynamics of monosynaptic response potentiation after nerve section at different distances from the spinal cord]. AB - The nature of increase in amplitude of monosynaptic reaction of the anterior root to stimulation of the posterior root was studied in 18, 24 and 48 hrs after preliminary cutting of the sciatic nerve in rats. After a close to spinal cord nerve cutting the amplitude of monosynaptic reaction on the side of the cutting increased within 18 hrs whereas after the distant cutting the monosynaptic reaction increase only occurred within 24 hrs. The increase in sensitivity to nerve impulses of the partially deafferented motoneurons is due not to the switching off the background impulsation but to the impairment of the nonimpulse influence on motoneurons. PMID- 6277703 TI - [Effect of lithium on the renal excretory function and Na, K-ATPase activity in rats]. AB - Single administration per os of the 0.066 M LiCl solution (3 ml per 100 g body weight) caused natriuresis. In the "lithium" rats with prolonged lithium treatment (i.p. injections during 10 days), polyuria developed; after water or 0.9% NaCl load excretion of Na, K, Ca, Mg and water increased as compared to the control rats. The saluretic effect was very obvious after 0.9% NaCl load in the "lithium" rats. The Na, K ATPase activity was measured by the quantitative cytochemical method, decreased only in the collecting ducts of the "lithium" rats. The natriuretic effect of acute lithium administration seems to be due to a lowering of the sodium proximal reabsorption. Prolonged lithium treatment suppresses the sodium transport in the collecting ducts. PMID- 6277704 TI - [Alpha 1- and beta 1-adrenergic reactions in acclimatization to cold]. PMID- 6277705 TI - [Oxygen, carbon dioxide and calcium control of the mechanisms of relaxation in the cerebral artery smooth musculature]. AB - The direct action of the hypoxic (PO2 60-20 mm Hg) and hypercapnic (PCO2 50-60 mm Hg) solutions on isolated strips of human cerebral arteries was studied. Verapamil, sodium nitroprusside, manganese, dichlordiphenyltrichlomethylmethan inhibited the hypoxic and hypercapnic relaxation. The data obtained suggest that lowering of PO2 or raising of PCO2 decrease the membrane calcium transport and increase the activity of the cellular calcium pump. PMID- 6277706 TI - Salt, soda and hydrogen peroxide ... is it enough? Yes! PMID- 6277707 TI - Salt, soda and hydrogen peroxide ... is it enough? No! PMID- 6277708 TI - Calcium movements and insulin release in pancreatic islet cells. AB - The mechanisms by which glucose provokes the intracellular accumulation of calcium in the B-cell were investigated by monitoring the efflux of 45Ca from preloaded and perifused rat pancreatic islets. Glucose provoked an initial fall followed by a secondary rise in 45Ca efflux. These two movements are sustained and reversible. They display distinct sensitivities towards glucose but both movements depend on the integrity of glucose metabolism in the islets. The secondary rise in 45Ca efflux reflects the rate of calcium influx through voltage sensitive calcium channels and corresponds to a process of calcium-calcium exchange. Glucose gates the calcium channels by increasing the endogenous generation of reduced pyridine nucleotides. The absence of extracellular sodium reduced the efflux of 45Ca and the inhibitory effect of glucose on calcium outflow. This suggests the existence in the islets of a process of sodium-calcium countertransport responsible for calcium extrusion from the B-cell. Glucose reduces 45Ca efflux by inhibiting this process. This inhibition may result from an increased endogenous generation of protons. In conclusion, glucose provokes an intracellular accumulation of calcium within the B-cell both by gating voltage sensitive calcium channels and by inhibiting sodium-calcium countertransport. PMID- 6277709 TI - New methods for analysing DNA make genetics simpler. PMID- 6277710 TI - Regulation of hexose transfer in giant muscle cells of the barnacle Balanus nubilus. PMID- 6277711 TI - Carriers and pores in mammary membranes. PMID- 6277712 TI - A general practice evaluation of selexid suspension as a treatment for acute urinary tract infection. AB - Fifty-seven general practice patients (fifty-five females and two males) with acute uncomplicated cystitis received a 5-day course of pivmecillinam suspension at a dose of 200 mg (2 sachets) three times a day. Positive bacteriological cultures were obtained from twenty-eight patients (49%) before treatment. The bacteriological cure rate was 100%. All patients experienced some relief of symptoms and thirty-nine patients (68%) were completely symptom-free after treatment. Selexid suspension was generally well tolerated. Side-effects were reported in only four patients(6%) and no patient had to stop treatment prematurely. PMID- 6277713 TI - Effect of the basal diencephalon on the development of Rathke's pouch in rats: a study in combined organ cultures. PMID- 6277714 TI - Effect of sulfation of CCK-8 on its stimulation of the endocrine and exocrine secretion from the isolated perfused porcine pancreas. AB - The secretory effect of sulfated and nonsulfated cholecystokinin octapeptide (CCK 8) was studied on the isolated perfused porcine pancreas. Both CCKs in concentrations from 10(-11) to 10(-8) mol/l in the presence of glucose (7.5, 5.0 or 3.5 mmol/l) were without effect on insulin and glucagon release. The exocrine secretion was stimulated by both CCKs in a dose-dependent manner, but sulfated CCK-8 was considerably more potent. The study shows that CCK-8, a major constituent of endogenous CCK, does not contribute to the incretin mechanism, irrespective of degree of sulfation. In contrast, CCK-8 is a potent stimulator of exocrine pancreatic secretion. For this effect sulfation is crucial. PMID- 6277715 TI - Calmodulin and insulin secretion. PMID- 6277716 TI - Cyclic AMP: a potent inhibitor of DNA synthesis in cultured arterial endothelial and smooth muscle cells. AB - The effect of dibutyryl cyclic AMP on DNA synthesis was studied in cultured human umbilical endothelial cells and rat aortic smooth muscle cells. Dibutyryl cyclic AMP (2 X 10(-4) mol/l) inhibited DNA synthesis in both arterial cell types when they were grown in medium supplemented with whole serum or with platelet poor serum, but had no effect in the absence of serum. An effect was seen one hour after the addition of the nucleotide, and the threshold concentration was between 2 X 10(-6) and 2 X 10(-5) mol/l. These results may have relevance to the interaction of platelets and insulin with the arterial wall in the development of atherosclerosis in diabetes. PMID- 6277718 TI - [Is aflatoxin B1 a hepatic carcinogen in man?]. PMID- 6277717 TI - Effects of hypophysectomy and growth hormone on cultured islets of Langerhans of the rat. AB - The effects of islet function of addition to the culture medium of rat growth hormone was studied in 4-day cultured islets of Langerhans from normal and hypophysectomised rats. In islets from hypophysectomised rats, rates of insulin release were 34% lower than in control rat islets; rates of insulin plus proinsulin and total protein biosynthesis were also lower by 48% and 16% respectively. The rates of glucose oxidation and the islet content of cyclic AMP were unchanged in islets from hypophysectomised rats but the islet content of calmodulin was decreased by 68%. The presence of rat growth hormone during the culture period restored the secretory response of hypophysectomised rat islets to that seen in control islets cultured without growth hormone but had only a marginal effect on the rate of insulin plus proinsulin biosynthesis, and no significant effect on islet calmodulin content. Glucose oxidation was increased by the presence of growth hormone during the culture period in both control (73% increase) and hypophysectomized (38% increase) rat islets. Addition of growth hormone to the culture medium also enhanced rates of insulin release and biosynthesis in control islets by 116% and 20% respectively. It is suggested that these changes arise primarily from modification of the synthesis of specific islet proteins. PMID- 6277719 TI - [Central alpha-adrenergic receptors are involved in the physiological regulation of exocrine pancreatic secretion; opiate receptors may not be]. PMID- 6277720 TI - [First case of so-called glucagonoma with amyloid stroma]. PMID- 6277721 TI - The role of opiate receptors and cholinergic neurons in the gastrocolonic response. AB - The aim of this study was to characterize the mechanism of the gastrocolonic response to eating in healthy human volunteers. A bipolar clip electrode recorded spike activity from the distal colon. A rapid increase in colonic spike activity occurred after eating a 1000-cal meal (16.1 +/- 2.8 spike potentials/10 min, p less than 0.001). The intravenous infusion of atropine inhibited the increase in postprandial colonic motility. Intragastric administration of procaine, a local anesthetic also inhibited the gastrocolonic response to eating. After sham feeding a modified 1000-cal meal, there was no increase in the gastrocolonic response. Morphine stimulated colonic spike activity (55.3 +/- 15.1 spike potentials/10 min, p less than 0.001). Naloxone (40 microgram/kg . h) completely inhibited morphine stimulation of the colon. Naloxone also inhibited the postprandial gastrocolonic response, but did not inhibit neostigmine stimulation of colonic motility. These data suggest: (1) the gastrocolonic response is mediated by afferent neural receptors in the gastroduodenal mucosa, (2) there is no cephalic phase in the gastrocolonic response to eating, and (3) efferent cholinergic neurons and opiate receptors are both necessary for the gastrocolonic response. PMID- 6277722 TI - Variability of motility of the ileum and jejunum in healthy humans. AB - Motor patterns of the distal small bowel were defined in healthy humans, using a multilumen polyvinyl tube, passed by mouth in 11 healthy subjects. Five recording sites, spanning 100 cm of tube and featuring a nitrogen hydraulic infusion system, were used to obtain records during 6 h of fasting and 6 or more hours after ingestion of a 600-kcal liquid test meal. The loci of recordings were designated as jejunal, ileal, or terminal ileal, as judged by the length of tube within the intestine and by fluoroscopy. During fasting, the migrating motor complex was present in all subjects and at all levels of the small intestine, but it could not be traced into the colon. Interdigestive cycles were defined primarily by the presence of an "activity front" (phase 3 of the migrating motor complex). Ninety-six migrating motor complexes occurred each 97 min (grand mean for all loci), but intervals between individual activity fronts varied markedly (15-195 min), in contrast to what is reported in other species. The velocity of aboral migration was 4.7 +/- 1.8, 1.3 +/- 0.4, and 0.9 +/- 0.2 cm . min-1 (mean +/- SD) in jejunal, ileal, and terminal ileal, respectively. Rates of continuous, rhythmic contractions during activity fronts declined distally: 12.5 to 10.5 (jejunal), 10.9 to 9.3 (ileal), and 10.0 to 8.6 cycle/min (terminal ileal), respectively. In individual subjects, maximum rates of contraction and velocities of migration always declined distally, but the duration of activity fronts was unrelated to the level of recording. Food interrupted the fasting cycles of motility for periods ranging from 2.75 to greater than 10 h. The transition from the fasting to the fed pattern was prompt and the postprandial motility was that of irregular bursts of contractions interspersed with transient quiescence. These studied demonstrate that the migrating motor complex occurs throughout the human small intestine, and that inter- and intraindividual variations are marked; food disrupts the complex for variable periods. These variations must be considered when abnormalities are being sought in disease states. PMID- 6277723 TI - Carcinoma of extrahepatic biliary system in an adult with cystic fibrosis. PMID- 6277724 TI - Regression of liver cell adenoma. A follow-up study of three consecutive patients after discontinuation of oral contraceptive use. AB - Three consecutive young female patients with unresected liver cell adenoma and a history of contraceptive use of several years' duration were followed up over a period of 2-4.5 yr after withdrawal of hormonal medication. One patient presented multiple adenomas and in another, the adenoma was associated with a Budd-Chiari syndrome due to bilateral thrombotic hepatic vein occlusion. Complete regression of the adenomas was documented in all 3 patients by ultrasonography, liver radionuclide scan, laparoscopy, or computerized tomography. It is concluded that, in selected cases, conservative management after withdrawal of hormonal contraception may be a valid alternative to surgical therapy. PMID- 6277725 TI - More on dietary fiber and carbohydrate metabolism. PMID- 6277726 TI - Verapamil--a potent inhibitor of esophageal contractions in the baboon. AB - Verapamil, a calcium channel blocking agent, has been shown to be a potent inhibitor of smooth muscle contraction. We studied the effects of verapamil on lower esophageal sphincter pressure and esophageal peristalsis in 5 awake baboons. An infusion of verapamil (80 micrograms/kg . min) decreased amplitude of peristalsis by 63% in the smooth muscle segments of the esophagus but had no effect on striated muscle. Duration of peristaltic waves in the smooth muscle portion of the esophagus was also shortened, but there was no effect on velocity of the contraction wave. Lower esophageal sphincter pressure decreased by 74% during verapamil infusion. These studies suggest that verapamil or other calcium channel blocking agents may be of potential use in the treatment of esophageal motility disorders associated with increased pressure. PMID- 6277727 TI - Stimulation of prostaglandin synthesis in rat cerebral cortex via a beta adrenoceptor. AB - 1. Synthesis of prostaglandin E2 (PGE2) and prostaglandin F2 alpha (PGF2 alpha) in rat cerebral cortex slices is increased by beta-adrenoceptor agonists. 2. Phenylephrine, an alpha-adrenoceptor agonist, has no effect while oxymetazoline increased PGF2 alpha only. 3. PG synthesis stimulation by noradrenaline (NA) was prevented by beta- but not by alpha-adrenoceptor antagonists. 4. Dibutyryl cyclic AMP and inhibitors of cyclic nucleotide phosphodiesterase augment PG synthesis. 5. Stimulation of PG synthesis in rat cerebral cortex by NA is mediated by a beta adrenoceptor. PMID- 6277728 TI - Biogenic amines and beta-endorphin responses in the rat vas deferens. AB - 1. Recent studies have demonstrated that activation of opioid peptide receptor by beta-endorphin results in a change of bioamines in peripheral tissues. 2. The present results could be interpreted assuming that the action of beta-endorphin, on the smooth muscle of the isolated rat vas deferens, is independent of catecholaminergic, cholinergic and tryptaminergic neurotransmission. 3. There is evidence that it is through a specific opioid 'epsilon-receptor' which have been shown a high degree of affinity for beta-endorphin. PMID- 6277729 TI - Stimulation of gastric enzyme secretion by porcine cholecystokinin in the ascidian Styela clava. PMID- 6277730 TI - In vitro study of frog (Rana ridibunda pallas) interrenal function by use of a simplified perifusion system. V. Influence of adrenocorticotropin upon progesterone production. PMID- 6277731 TI - The in vitro response of the trout interrenal to various fragments of ACTH. PMID- 6277732 TI - The responses of the Malpighian tubules of Locusta to hormones and other stimulants. PMID- 6277733 TI - Evolutionary relationships among five subspecies of Mus musculus based on restriction enzyme cleavage patterns of mitochondrial DNA. AB - The intra- and intersubspecific genetic distances between five subspecies of Mus musculus were estimated from restriction enzyme cleavage patterns of maps of mitochondrial DNA (mtDNA). The European subspecies, M. m. domesticus and Asian subspecies, M. m. bactrianus, M. m. castaneus, M. m. molossinus and M. m. urbanus were examined. For each subspecies, except M. m. urbanus, at least two local races from widely separated localities were examined. Intrasubspecific heterogeneity was found in the mtDNA cleavage patterns of M. m. bactrianus and M. m. castaneus. M. m. molossinus and M. m. domesticus, however, revealed no intrasubspecific heterogeneity. Four of the subspecies had distinct cleavage patterns. The fifth, M. m. urbanus, had cleavage patterns identical to those of M. m. castaneus with several enzymes. Estimates of genetic distances between the various races and subspecies were obtained by comparing cleavage maps of the mtDNAs with various restriction enzymes. Nucleotide sequence divergences of mtDNA between local races were estimated to be less than 0.4% in M. m. bactrianus and less than 0.3% in M. m. castaneus. The times of divergence of both subspecies were calculated to be 0.1--0.2 x 10(6) years. These values suggest that the intrasubspecific divergence began some 0.1--0.2 x 10(6) years ago. On the other hand, nucleotide sequence divergences between European subspecies M. m. domesticus and Asian subspecies M. m. bactrianus and M. m. castaneus were 7.1% ane 5.8%, respectively. The times of divergence were calculated to be 2.1--2.6 x 10(6) years. Further, the nucleotide sequence divergence and time of divergence between M. m. molossinus and the other two Asian subspecies were comparable to those between M. m. molossinus and M. m. domesticus (about 3% and 1 x 10(6) years, respectively). These results suggest that M. m. molossinus is situated in a unique evolutionary position among Asian subspecies. PMID- 6277734 TI - Construction of new vectors for cloning of promoters. AB - Vectors for cloning promoter-DNA fragments were derived from plasmid pBR313 (Bolivar et al., 1977). These have several unique restriction sites and carry the trpA gene from Escherichia coli as a selective marker. The selection is based on an enhancement of the growth rate of those bacteria in which the expression of trpA is directed by the cloned promoter. The expression of trpA can be determined quantitatively, independently of the copy number of the vector, and should reflect the apparent strength of the promoter, since the DNA segment located before trpA contains translational stop signals in all three reading frames. PMID- 6277735 TI - Structure, stability, methylation, expression and glucocorticoid induction of endogenous and transfected proviral genes of mouse mammary tumor virus in mouse fibroblasts. AB - A sequence of mouse genomic DNA containing an endogenous mouse mammary tumor virus (MMTV) provirus, which was isolated by molecular cloning in lambda vector phage, has been reintroduced into cultured mouse L cells, using the thymidine kinase cotransfection technique. Individual cell clones that acquired the transfected MMTV proviral DNA have been isolated. The transfected DNA remains stable in these cells and does not become methylated. It is transcribed into MMTV RNA, And the transcription is stimulated by glucocorticoid hormones. PMID- 6277736 TI - Versatile cloning vectors derived from the runaway-replication plasmid pKN402. AB - Two cloning vectors have been constructed employing runaway-replication mutants of plasmid R1. One of these, pMOB45, carries tetracycline and chloramphenicol resistance. The other, pMOB48, carries chloramphenicol resistance, lacOP, and an assayable part of the lacPOZ operon. Both of these plasmids can be amplified to high levels by heat induction, which condition does not lead to inhibition of protein synthesis; thus the plasmid can produce large amounts of DNA and protein. In pMOB48, a unique BamHI site is present near the amino-terminus of the beta galactosidase gene. Chimeras formed by the insertion of restriction fragments at this site can be detected on X-gal plates, and can be used for the lacIq controlled expression of proteins which are fused to the amino-terminus of beta galactosidase. Induction with IPTG at 40 degrees C leads to the synthesis of extremely high levels of proteins whose gene have been cloned into this site. PMID- 6277737 TI - Region- and strand-specific mutagensis of a recombinant plasmid. AB - Techniques were developed to mutagenize a single DNA strand in a specific region of the tetracycline-resistance (tetr) gene of the plasmid pKB280 that also carries the lambda repressor gene. Separate annealings of complementary single strands gave two isomeric, circular plasmids containing a 275-nucleotide, single stranded region (gap) in the tetr gene. One of the isomeric, gapped plasmids was mutagenized specifically with sodium bisulfite such that an estimated 98% of the molecules had suffered at least one C to U conversion in the gap. The mutagenized gap was filled in with DNA polymerase. These molecules transformed Escherichia coli strain MM294 to lambda-immunity with the same frequency as unmutagenized, gap-filled pKB280. Of the lambda-immune transformants, 32% were Tcr and 68% were Tcs. Restriction analysis of plasmids from some Tcs transformants showed losses of restriction sites within the gap and at the gap termini, but none outside the gap. No deletions were detected. PMID- 6277739 TI - Characterization of coliphage lambda hybrids carrying DNA fragments from Herpes simplex virus type 1 defective interfering particles. AB - We describe the characterization of 34 hybrid lambda bacteriophages carrying EcoRI fragments obtained from DNA of defective interfering particles of the Patton strain of Herpes simplex virus type 1 (HSV-1). All cloned fragments contained S region terminal repeat sequences (TRs) fused to unique HSV-1 DNA. Several fragments contained deletions and rearrangements not described previously for DNA of HSV-1 defective interfering particles. A model describing the generation of defective interfering DNA based on recombination events involving the terminal "a" sequence as presented. PMID- 6277738 TI - Integration of foreign genome fragments into cells transformed or cotransformed with fragmented adenoviral DNA. AB - The integration of DNA of highly oncogenic simian adenovirus type 7 (SA7) and non oncogenic human adenovirus type 6 (Ad6) into the genome of newborn rat kidney cells transformed by fragmented DNA preparations was studied using reassociation kinetics and spot hybridization. Transforming DNA was fragmented with the specific endonuclease SalI (SA7) and BglII (Ad6). In contrast to the cell transformation by intact viral DNA, transformation by fragmented DNA resulted in integration into the cellular genome of not only the lefthand fragment with the oncogene but also of other regions of the viral genome. Additionally integrated fragments were stable and preserved during numerous passages of cells lines, although they were no expressed, at least in the case of the Ad6-transformed cell line. The integration of the fragments of SA7 DNA was accompanied by loss of 25 50% of the mass of each fragment. Adding the linear form of the pBR322 plasmid to the preparation of transforming Ad6 DNA also contributed to its cointegration into the genome of the transformed cell. This technique of cell cotransformation with any foreign DNAs together with the viral oncogens may be used as an equivalent of an integration vector for eukaryotic cells. PMID- 6277740 TI - Cloning a promoter that puts the expression of tetracycline resistance under the control of the regulatory elements of the mer operon. AB - We have sub-cloned from the Eco RI-H fragments of the IncFII plasmid R100 a 260 bp EcoRI fragment, using the promoter-cloning vehicle, pBRH4, (The Inc FII plasmid codes for the mer operon, and pBRH4 expresses tetracycline resistance only when the deleted tet promoter has been replaced by another sequence that can serve as a promotor). With the 260-bp fragment inserted, the derivative plasmid, pFB4, directs the expression of tetracycline resistance only if there is a second plasmid in the strain that carries the merR-positive regulatory element. Under these conditions, the level of tetracycline resistance is directly proportional to the concentration of Hg2+ present in the medium. The 260-bp fragment also allows low-level constitutive expression of tet resistance when transactivated with merR mutants that have a "micro-constitutive" phenotype. The 260-bp mer promoter fragment contains a single HincII site; there is also but one HincII site in the EcoRI-H fragment of R100 from which the promoter fragment was derived. Restriction analysis of purified Eco RI-H DNA shows that the single HincII site is at 550 bp from the "right"terminus of the IS1b element, which is also present in the EcoRI-H fragment. Because of its biological activity and its location within the "H" fragment, this promoter is very likely a promoter for the structural genes of the operon. PMID- 6277741 TI - Insulin insensitivity and hyposuppressibility of glucagon by hyperglycemia in aged Wistar rats. AB - Insulin and glucagon secretion after glucose load and the hormone-receptor inter action were compared between 6- and 24-month-old Wistar rats. The 24-month-old rats showed a failure to decrease glucagon secretion after glucose load in contrast to 6-month-old rats with a similar degree of glucose tolerance and insulin response. Age-related change of insulin receptor of rat livers was tentatively suggested to be a disappearance of the binding site with higher affinity. Affinity of the glucagon receptor did not differ significantly between the two groups. The hyposuppressibility of glucagon by hyperglycemia and the decrease of insulin affinity might contribute to the glucose intolerance with aging. PMID- 6277742 TI - Childhood cancer: triumph over tragedy. Introduction. PMID- 6277743 TI - Management of children with Wilms' tumor: defining the risk-benefit ratio. PMID- 6277744 TI - [Percutaneous 239Pu uptake in rats with skin burns from nitric acid]. PMID- 6277745 TI - [Quantitative determination of enteroviruses in water]. PMID- 6277746 TI - Nutrition: why eat a fiber-rich diet? PMID- 6277747 TI - Triggering of the finger at the flexor retinaculum. AB - An unusual case of trigger finger associated with paraesthesia of the hand is described. The site of triggering was at the flexor retinaculum consequent upon a tumour of the profundus tendon. Nerve conduction study of the median nerve revealed a co-existing incipient median neuritis. PMID- 6277748 TI - Giant cell tumour of metacarpal in a child of unusual site and age. AB - The opportunity of treating a giant cell tumour in a metacarpal of a six year old girl prompted the revision of the literature on this subject, verifying the extraordinary rarity of this kind of tumour in the bones of the hand and in children. The affected bone was excised and a metatarsal of the foot of the same girl transplanted in its place. PMID- 6277749 TI - [Successful treatment of severe refractory hypertension with captopril]. PMID- 6277750 TI - [The interaction of hormones and the exocrine pancreas]. PMID- 6277751 TI - [Pharmacological studies on antispasmodics. V. Effect of 3-(di-2 thienylmethylene)-5-methyl-trans-quinolizidinium bromide (HSR-902) on the junctional transmission of pelvic nerve ending in isolated urinary bladder (author's transl)]. AB - Effects of HSR-902, a new antispasmodic agent, on the junctional transmission of pelvic nerve ending in isolated urinary bladder was compared with those of atropine sulfate, butylscopolamine bromide, timepidium bromide and prifinium bromide. Tetrodotoxin (3 x 10(-8)g/ml) completely inhibited the contraction of isolated guinea pig urinary bladder induced by transmural stimulation (TM contraction), but hexamethonium chloride (1 x 10(-4)g/ml) and atropine sulfate (1 x 10(-5)g/ml) exhibited no inhibition and a slight inhibition, respectively. Neostigmine bromide (1 x 10(-7)g/ml) increased TM contraction, and the effect was completely inhibited by atropine sulfate (1 x 10(-5)g/ml). Antispasmodic agents (1 x 10(-7) - 1x 10(-5)g/ml), excluding HSR-902, showed slight inhibition on TM contraction. In contrast, HSR-902 (1 x 10(-6) - 1x10(-5)g/ml) increased TM contraction and/or resting tone (1 x 10(-5)g/ml). Phenoxybenzamine hydrochloride (1 x 10(-8)g/ml) and tolazoline hydrochloride (1 x 10(-5)g/ml), alpha-blocking agents, increased both TM contraction and resting tone. Increasing effects of HSR 902 on TM contraction and/or resting tone were completely inhibited by noradrenaline (1 x 10(-5)g/ml) with propranolol hydrochloride (1 x 10(-5)g/ml). These results suggested that these antispasmodic agents scarcely inhibited the junctional transmission in postganglionic cholinergic nerve ending induced by transmural stimulation of isolated guinea pig urinary bladder, and HSR-902 rather increased the transmission. Since it was also suggested that there was alpha adrenoceptor inhibiting acetylcholine-release in the postganglionic cholinergic nerve terminal, the transmission increasing action of HSR-902 would be due to its alpha-blocking action. PMID- 6277752 TI - [Effects of anesthesia on the levels of cyclic nucleotides in rat brain regions (author's transl)]. AB - The following 4 Wistar rat groups were sacrificed by microwave irradiation: 1) hypothermia-rebreathing anesthesia (kept 3 hr at 4 degrees C in a 4 1 box), 2) pentobarbital anesthesia (50 mg/kg, s.c., 1 hr after), 3) reserpinized rats (5 mg/kg/day, s.c. for 3 days), and 4) nontreated control rats. Cyclic nucleotides and noradrenaline were analyzed in seven brain regions: cerebellum, pons and medulla oblongata, hypothalamus, mid brain, striatum, hippocampus, and cerebral cortex. In most part of the brain regions, the levels of cyclic AMP, cycli GMP and noradrenaline had a tendency to decrease after the treatment by hypothermia rebreathing or pentobarbital as compared with control animals. The levels of cyclic AMP in the cerebellum and cyclic GMP in the hypothalamus were not affected by these anesthesia. After reserpinization, the level of cyclic GMP markedly elevated in all regions of the brain except in the cerebellum. In all brain regions of the control and the hypothermia-rebreathing groups, the coefficients of correlation between cyclic AMP and noradrenaline, cyclic GMP and nonadrenaline, cyclic AMP and cyclic GMP were positive, negative and negative, respectively. The cyclic AMP/cyclic GMP ratio in most of the brain regions decreased in the reserpinized group, and increased in the hypothermia-rebreathing group as compared to the control and the pentobarbital groups. From these results, it is postulated that some other neurons, not only monoaminergic neuron, regulate the level of cyclic nucleotides, and that the different sensitivity of these neurons to the anesthesia brings about various effects on the metabolism of cyclic nucleotides in each brain region. PMID- 6277753 TI - [Modulatory actions of prostaglandins in autonomic neuro-effector transmissions (author's transl)]. AB - In addition to the negative feedback control of transmitter release exerted by the neurotransmitter itself, there are also other substances which may play a modulatory role in autonomic neuro-effector transmissions. Prostaglandins (PGs) belong to this class of substances. This review highlights the action of PGs and indomethacin on adrenergic and cholinergic neuro-effector transmissions in the guinea-pig vas deferens and dog trachea. In the guinea-pig vas deferens, low concentrations of the PGE series markedly suppressed the amplitude of e.j.p. without affecting the membrane potential, input membrane resistance or sensitivity of the smooth muscle cells to noradrenaline. [Ca]0 appeared to counteract the inhibitory action of PGs on the amplitude of e.j.p., indicating that low concentrations of PGs interact with [Ca]0 at the activated nerve terminals. Endogenous PGs may not play, however, a physiological role in the regulation of noradrenaline release, since indomethacin had no effect on the e.j.p. On the contrary, gradual and continuous reduction in the amplitude of e.j.ps was abolished by indomethacin in the dog trachea and e.j.ps with a constant amplitude were recorded. Low concentrations of PGs markedly reduced the amplitude of e.j.ps without affecting membrane properties of the smooth muscle cells. When indomethacin in repeated doses was given, spontaneous coughing and wheezing occurred in half the number of dogs. Endogenous PGs therefore, probably play an important role in the negative feedback regulation of transmitter release from the cholinergic nerve terminals, in the dog trachea. Topical and species differences in the modulatory role of PGs was discussed. PMID- 6277754 TI - Changes in the cytochrome c content during the aerobic adaptation of Paracoccus denitrificans. AB - Concentration of cytochrome c decreases when shaking anaerobically grown cells in a non-growth medium down to 50% of the original amount in the cell, depending on the degree of aeration. Only 10-20% of this amount can be found in the adaptation medium. The main portion of cytochrome c is degraded during the adaptation. Inhibitors of proteinases do not influence the degradation. Addition of mammalian cytochrome c fully prevents the degradation of bacterial cytochrome c but not its release from cells. Potassium hexacyanoferrate(III) exhibits a similar effect as oxygen. The degradation system is probably localized in the periplasmic space of the cells. PMID- 6277755 TI - Nonspecificity of some commonly used inhibitors of DNA synthesis in cultures of Streptococcus faecalis. AB - The specificity of three commonly used inhibitors of DNA synthesis were tested in the batch culture of Streptococcus faecalis ATCC 8043 in rich broth medium. It was shown that nalidixic acid, mitomycin C and 6-(4-hydroxyphenylazo)uracil inhibit the growth of the cell mass as much as they decrease net DNA synthesis. Hence the drugs tested are highly unspecific inhibitors of DNA synthesis in S. faecalis; i.e. they all interfere with other processes as well as with DNA synthesis. PMID- 6277756 TI - Effects of drugs which increase adenosine 3',5'-monophosphate content on rabbit intestinal smooth muscle. PMID- 6277757 TI - 18-hydroxy-11-deoxycorticosterone response to ACTH, insulin and furosemide administration in essential hypertensive patients. AB - Investigations were carried out on the behavior of 18-hydroxy-11 deoxycorticosterone (18-OH-DOC) in essential hypertension (EH) under exogenous administration of synthetic ACTH and insulin. 40 stable EH patients and 21 normal subjects were included in the study. The increase (12-fold basal values) in plasma 18-OH-DOC in normal subjects under Tetracosactide was significantly higher than cortisol (4-fold basal values). Furthermore, insulin hypoglycemia increased 18-OH-DOC levels 5-fold, whilst basal values of cortisol were increased 2-fold. An increase in 18-OH-DOC and cortisol was also observed in EH patients: in the subgroup with normal and low plasma renin activity, however, the rise in these two steroids was significantly lower than in normal subjects both under Tetracosactide and insulin. No significant hormonal modifications were observed after furosemide administration either in the normal subjects or in the EH patients. 18-OH-DOC by itself does not, therefore, appear to play a pathogenetic role in EH. PMID- 6277758 TI - In vivo distribution of radioiodinated ACTH1-24 in the rat. AB - After intravenous injection of 125I-ACTH1-24 into rats, the highest concentration of 125I was found in kidneys, adrenal and liver. Addition of 5- and 30-fold excess unlabelled ACTH reduced the uptake of 125I by 50 and 68%, respectively, indicating that the adrenal uptake was specific. Pretreatment with dexamethasone decreased the adrenal uptake of 125I and caused adrenal atrophy. Chronic ACTH treatment increased the size of the adrenals, but did not affect the adrenal uptake of 125I. These experiments demonstrate selective uptake of 125I by the adrenals after administration of 125I-ACTH1-24. PMID- 6277759 TI - The effect of oxidation of the Met27 residue of [125I]monoiodoglucagon on receptor-binding affinity. AB - When glucagon is iodinated by the chloramine-T method, the Met27 residue is oxidized. This is not the case when the iodination is performed by the lactoperoxidase method. The two preparations can be purified to the same specific activity using QAE-Sephadex A-25 ion exchange chromatography. Receptor-binding studies in isolated rat adipocytes or hepatocytes revealed that the oxidized from possessed an average-binding affinity which is only about two thirds of that of the non-oxidized form. The reduced affinity of the oxidized tracer cannot be explained by an increased rate of dissociation. PMID- 6277760 TI - The ACE test for sarcoidosis: when to use, how to interpret. PMID- 6277761 TI - Physiology and pharmacology of antiarrhythmic drugs. PMID- 6277762 TI - Modern aspects of the morphology of viral hepatitis. AB - The morphologic pathology of human viral hepatitis and its sequelae are reviewed in this article. Emphasis is placed on new information, including the current status of the pathologic diagnosis of hepatitis non-A, non-B. The article includes a discussion of aspects of the virology that are pertinent to an understanding of the significance of viral markers in the liver. A small contribution of the authors is a brief description of the neocholangiole, a duct of Hering-like structure seen following hepatic necrosis from many causes, including the severe forms of viral hepatitis. PMID- 6277763 TI - Primary hepatocellular carcinoma--etiology, pathogenesis, and prevention. AB - Worldwide, primary hepatocellular carcinoma may be the most common cancer in men. Usually it occurs in association with cirrhosis or chronic hepatitis. In this article I present evidence that the majority of these cases are associated with, and probably caused by, persistent infection with hepatitis B virus. I also propose a model that describes the role of the hepatitis B virus in the pathogenesis of primary hepatocellular carcinoma. If persistent infection with hepatitis B virus is required for the development of most cases of primary hepatocellular carcinoma, prevention of such infections should prevent most cases of primary hepatocellular carcinoma. A vaccine against hepatitis B virus, prepared from surface antigen particles (HBsAg) harvested from the blood of hepatitis B virus carriers, has been safe and effective in preventing acute infection with hepatitis B virus in adult homosexual men. If it is equally safe and effective in young children, it will be administered widely to populations living in areas endemic for hepatitis B virus infections. Thereafter it may be possible to learn within five to 10 years whether the vaccine prevents chronic liver disease. If it does, it will be reasonable to conclude that it will also prevent primary hepatocellular carcinoma. PMID- 6277764 TI - Hepatitis A virus: virologic, clinical, and epidemiologic studies. AB - The last decade has borne witness to accelerated expansion of our understanding of hepatitis A virus. The agent of type A hepatitis is an RNA virus with a mean diameter of 27 nm. and biochemical-biophysical properties of an enterovirus. A variety of sensitive specific serologic techniques have been developed with which to identify hepatitis A virus and antibody, and both chimpanzees and marmosets have been studied extensively as experimental animal models. As a result of these studies, in vitro cultivation of hepatitis A virus has finally been accomplished, and a commercial radioimmunoassay for IgM antibody to hepatitis A virus has been developed for the rapid diagnosis of hepatitis A virus infection during acute illness. Clinically the illness caused by hepatitis A virus is relatively mild, often subclinical, and of limited duration and does not progress to chronic liver disease. This relative clinical benignity is reflected, according to preliminary histologic observations, in the sparing of the centrozonal area of the liver lobule. Rarely, however, hepatitis A virus can cause fulminant hepatitis. Type A hepatitis is transmitted almost exclusively by the fecal-oral route, and its spread is enhanced by epidemiologic settings favoring dissemination of enteric infections. Hepatitis A virus does not contribute to transfusion associated or other types of percutaneously transmitted hepatitis. Exposure to the virus increases as a function of age and decreasing socioeconomic class, but the incidence of hepatitis A virus infection in urbanized societies is decreasing. There is no evidence for the existence of chronic hepatitis A virus carriage; natural perpetuation of hepatitis A virus in urban communities appears to depend on a reservoir of nonepidemic, clinically inapparent cases. Until a vaccine, now being developed, becomes available, prevention of hepatitis A virus infection will continue to depend on maintenance of high standards of environmental and personal hygiene and on timely administration of immune serum globulin. Such prophylaxis may confer long lasting passive-active immunity but more frequently prevents infection entirely. PMID- 6277765 TI - Population, formal genetics, and linkage relations of the phosphoglycolate phosphatase (PGP)--E.C.3.1.3.18. AB - One hundred and sixty-seven blood donors, 26 families with 72 offspring and 12 motherchild couples were studied for the phosphoglycolate phosphatase polymorphism. In hemolysates, the isozymes are stable for at least five weeks. The distribution of observed phenotypes in the population study did not diverge from the expected values according to Hardy-Weinberg law. In the family study, the formal genetic model of three alleles--PGP1, PGP2 and PGP3 at one autosomal locus-could be confirmed. Among 33 individuals from a Mongoloid population PGP1 was observed in 100%. This observation lead us to the conclusion, based also on recent data in Negroid populations (Barker and Hopkinson 1978), that phosphoglycolate phosphatase may be a more recent polymorphism of Caucasoid populations. Linkage studies with the hp alpha locus an chromosome 16 resulted in 19 meiotic divisions of 4 informative families in a lod score peak of 0.23 at theta 0.25 being inconclusive. The inclusion of the PGP system in paternity testing is also discussed. PMID- 6277766 TI - Hitchhiking and linkage disequilibrium between hemoglobin S and nearby restriction sites. AB - The association of the hemoglobin S mutation with an unusual 13 kb Hpa I restriction fragment has been reexamined using new data and a stochastic model for linkage disequilibrium. In agreement with earlier analyses with deterministic models, the present-day association of the beta S allele with the 13 kb fragment is lower than would be expected if all beta S alleles carried by American blacks were derived from a unique mutation that occurred in the ancestral Bantu population. However, from the stochastic model, if the effective population size was small for an appreciable period during the time that selection has been operating on HbS, then the confidence limits on the allelic association are very large and a unique West African beta S mutation cannot be ruled out from the two locus data alone. PMID- 6277767 TI - Evidence for the existence of a phosphoinositol kinase in chicken erythrocytes. PMID- 6277768 TI - Enzymes of ethanolamine plasmalogen metabolism in neuron- & glial-enriched fractions of rat brain. PMID- 6277769 TI - Intracellular localization of starch phosphorylase from banana (Musa paradisiaca) leaves. PMID- 6277770 TI - Fructose-1,6-bisphosphatase & myo-inositol synthase: a phylogenetic search. PMID- 6277771 TI - Aminoglycosides and polymyxin B as inhibitors of antigen attachment to erythrocytes for agglutination by Salmonella antibodies. AB - Previous investigations indicate that polymyxin B inhibits attachment of lipopolysaccharides (O antigens) to erythrocytes and subsequent hemagglutination by the corresponding bacterial antibodies. In this study the effect of this antibiotic was compared to that of the aminoglycosides gentamicin, tobramycin, and amikacin. Using both crude O antigens and highly purified lipopolysaccharides, polymyxin B was at least 100 times more effective than the aminoglycosides. Amikacin proved to be less effective than gentamicin and tobramycin. PMID- 6277773 TI - Specific effects on human neutrophils of antibodies to a membrane protein constituent of neutrophil receptors for chemotactic formyl-methionyl peptides. PMID- 6277772 TI - Antigen-induced arthritis: inflammation and antigen handling after two different doses of intra-articularly injected antigen. AB - We studied (i) the inflammatory response induced by intra-articular (i.a.) injection of 2.5 and 0.5 mg 125I-labelled bovine serum albumin (125I-BSA) into the paired knee joints of rabbits immunized with BSA in Freund's complete adjuvant, and (ii) handling of these different doses of antigen by means of regular external radioactivity measurements. The joint inflammation was quantified by 99mTechnetium pertechnetate uptake measurements. More severe arthritis was seen in the 2.5 mg BSA-injected than in the paired 0.5 mg BSA injected knee, both in the early and in the late phase of antigen-induced arthritis. External radioactivity measurements showed enhanced disappearance of radioactivity from the 2.5 mg 125I-BSA challenged knee joint, resulting in lower percentual retention of the dose injected than in the 0.5 mg injected knee joint. However the calculated absolute amount of long term retained BSA in the 2.5 mg BSA injected knee, 4 weeks after i.a. injection, was still about 2.8 times the quantity of BSA retained in the 0.5 mg BSA-injected paired knee joint. These data indicate that the severity of both the early and late phase of antigen-induced arthritis is i.a. antigen dose-dependent and, in addition, suggest that handling of i.a. antigen depends in part on the severity of joint inflammation. PMID- 6277774 TI - Immunization of mouse spleen cell cultures in the absence of serum and its proteins using SiO2 and 2-mercaptoethanol. AB - High recoveries of plaque-forming cells were obtained in a serum-free medium using 2-mercaptoethanol and SiO2. Under such conditions an increase in the number of plaque-forming cells occurred between the fourth and fifth day of incubation, as compared with cultures with foetal calf serum. The addition of glutathione to the medium had a stimulating effect, it was however not absolutely necessary for a positive response. The recovery of plaque-forming cells was strongly influenced by an interdependence of cell density and amount of SiO2 gel particles. PMID- 6277776 TI - Identification of a serum-derived promotor of granulocyte granule secretion: study on a patient with chronic pruritus. PMID- 6277775 TI - Modulatory role of cyclic AMP in the release of platelet-activating factor from human polymorphonuclear leucocytes. AB - Zymosan coated with complement (Zc) was observed to induce a transient elevation of the intracellular cyclic AMP in human polymorphonuclear cells: a two- to three fold increase was observed within 1 min after stimulation and approached prestimulation levels by 2 min incubation. These changes in cyclic AMP were not associated with significant changes in cyclic GMP levels. Zymosan caused the release of PAF and beta-glucuronidase and particle uptake, which was initiated about 5 min after stimulation. These results suggest that the transient increase in cyclic AMP content might regulate an early event during mediator release. In an attempt to study further the significance of this rise in cyclic AMP, cells were preincubated with various phosphodiesterase inhibitors. Preincubation of the cells with methylisobutylxanthine (MIX, 10(-6) M to 5 X 10(-5) M), theophylline (3 X 10(-5) to 3 X 10(-3) M) or dipyridamole (10(-6) M to 10(-4) M) enhanced the increase in cyclic AMP levels, but resulted in dose-dependent inhibition of Zc induced mediator release. Particle uptake and beta-glucuronidase release were less sensitive than PAF release to phosphodiesterase inhibitors, which argues in favour of the independence of both phenomena. Synergistic experiments with MIX and cyclic AMP indicate that the effect of this drug is through its action on cyclic AMP levels. These results suggest that while Zc-induced cyclic AMP elevation might occur in an intracellular place critical to its effect; phosphodiesterase inhibitors may elevate cyclic AMP levels throughout the cell and therefore inhibit the biological response. PMID- 6277777 TI - Inability of tumour cells to elicit the respiratory burst in cytotoxic, activated macrophages. AB - Activated macrophages from Corynebacterium parvum-treated mice are cytotoxic to non-antibody-coated tumour cells and have an augmented respiratory burst potential when compared to resident macrophages. We have investigated the possible involvement of the respiratory burst as an effector mechanism in this type of tumour killing. Scavengers of toxic metabolites of oxygen such as catalase, superoxide dismutase, 2,3-dihydroxybenzoate, ethanol, and cytochrome c did not inhibit macrophage cytotoxicity in this system. To investigate whether or not neoplastic cells stimulate the macrophage respiratory burst, we exposed activated macrophages to viable tumour cells and monitored macrophage superoxide anion production, chemiluminescence, and hexose monophosphate shunt activity. None of these indicators of the macrophage respiratory burst was stimulated by the tumour cells towards which the macrophages were cytotoxic. The data suggest that the macrophages burst is not utilized as an effector mechanism in the non antibody-mediated macrophage tumour cytotoxicity reaction. PMID- 6277779 TI - Lipolytic and enkephalinase inhibitory activities of polypeptides from placental autolysate prepared for therapeutic purposes. AB - In order to study the pharmacodynamic basis of therapeutic uses of placental extracts researches was carried out on Sephadex G25 factors of autolytic extract of human placenta. Four of these fractions showed immunoreactivity for both beta endorphin and ACTH, and were able to stimulate lipolysis in rabbit adipose tissue. An inhibition of the enkephalinase activity of rat brain homogenates was displayed by placental fractions of low molecular weight. These results suggest that beta-endorphin and ACTH play a roles in the pharmacological activity of placental extracts. PMID- 6277778 TI - 5-Nucleotidase in liver injury induced by heavy metals. A histochemical study. AB - The alterations in the distribution of 5-nucleotidase being the indicator of the functional state of liver parenchyma have been determined histochemically in the rat (Rattus rattus albino) after experimental poisoning with a few heavy metals viz. lead, mercury, cadmium, molybdenum, copper, and zinc that possess considerable environmental and nutritional significance. Present results show that all these metals affected the distribution and activity of 5-nucleotidase that has been explained on the basis of binding affinity of these metals, state of plasma membrane, resorption of nucleotides and variations in the level of enzyme protein. Presumably, all these factors have implications for the interpretation of observed changes in enzyme activity and are specific to each metal. PMID- 6277780 TI - Fibronectin interacts with Clq, a subcomponent of the first component of complement. AB - Human Clq, a subcomponent of the first component of complement interacts with human fibronectin. Using ELISA methodology fixation of Clq to solid phase fibronectin, as well as fibronectin to solid phase Clq has been demonstrated. Cl in its native macromolecular form displays little reactivity for fibronectin, nor does Cl reconstituted from Clq, Clr and Cls in the presence of Ca2+ ions. Heating of Clq above its thermal transition temperature (51 degrees C) induces an increased binding capacity for fibronectin. On the other hand, a mixture of the dissociated A, B and C chains of Clq is less active than native Clq. The binding of fibronectin appears to be mediated by the A chain. Studies with Clq deprived of its globular parts by peptic digestion indicate that the collagen-like regions of Clq are involved in fibronectin binding. In contrast, collagenase treatment of Clq abrogates its fibronectin binding capacity. PMID- 6277782 TI - Cholesterol induced changes in concanavalin A agglutinability of Entamoeba histolytica. PMID- 6277781 TI - A study of presynaptic alpha-adrenoceptors of rabbit intestine. PMID- 6277783 TI - Glucose-6-phosphatase in human amniotic fluid. PMID- 6277785 TI - Correlation between growth potential of mouse hepatitis viruses in macrophages and their virulence for mice. AB - Correlation between the virulence of mouse hepatitis virus (MHV) for mice and the growth potential of the virus in peritoneal adherent cells was observed for highly virulent MHV-2 and avirulent MHV-1, JHM, and MHV-S strains. However, this phenomenon was not observed in strain MHV-3, which multiplied to almost the same degree in peritoneal adherent cells from susceptible and resistant mouse strains. PMID- 6277784 TI - Independence of H-2 and viral antigens on the cell surface and absence of H-2 antigens on murine leukemia virus and mouse mammary tumor virus particles. AB - By indirect immunoelectron microscopy we tested for the presence of H-2 antigens on murine mammary tumor virus (MMTV) and murine leukemia virus (MuLV) particles. The association of H-2 antigens and viral antigens on the virus-infected cell surface was investigated with antibody-induced redistribution. Mammary tumor cells and leukemia cell lines with different H-2 genotypes and carrying different MuMTV or MuLV were used. No H-2 antigens could be demonstrated on the envelope of MMTV and MuLV particles, even after the permeabilization of their envelopes with saponin. On the surface of virus-infected cells antibody-induced patching or capping of the viral antigens did not result in copatching or cocapping of the H 2 antigens. In the reciprocal tests no co-redistribution of viral antigens with H 2 antigens was seen. Our experiments failed to show any physical association between H-2 antigens and MMTV or MuLV antigens on the cell surface. PMID- 6277786 TI - Analysis of age-dependent resistance to murine coronavirus JHM infection in mice. AB - Resistance to intraperitoneal murine coronavirus JHM infection in mice develops with age. C3H mice were found to be fully susceptible up to the age of 20 days and resistant after 23 days of age. Protection of susceptible animals from death due to infection could be achieved by maternal antibodies or by transfer of spleen cells from immunized, but not from nonimmunized, donor mice. Lack of protection by transfer of unprimed adult spleen cells was not related to immunosuppression by the host. Moreover, resistance of adult mice could not be abrogated by application of lymphocytes from suckling mice, although immune suppression by other means did affect the resistance of adult animals. On the other hand, spleen cells from nonimmunized mice could be primed with inactivated JHM virus in suckling mice and protected these mice from death due to a subsequent virus infection. Thus, the outcome of infection with JHM virus in suckling and adult mice can be influenced by immunological events, but is not exclusively due to the different stages of immune competence. PMID- 6277787 TI - Pathogenesis of Herpes simplex virus infections in guinea pigs. AB - The pathogenesis of herpes simplex virus types 1 and 2 has been studied in guinea pigs after inoculation by various routes (subcutaneous and intradermal infection in footpads and vaginal infection). Clinical observations as well as virus isolation studies are reported. Herpes simplex virus type 2 infection by all three routes of inoculation led to acute primary and recurrent lesions. Virus persisted in the nervous system, particularly in sensory ganglia, and locally at the site of inoculation. Herpes simplex virus type 1 infection induced no or very mild primary symptoms. Recurrent lesions were only observed after intradermal inoculation. Invasion of the nervous system and consequent establishment of latent ganglionic infection was less efficient than after herpes simplex virus type 2 infection. Peripheral persistence was, however, equally common. PMID- 6277788 TI - Monoclonal antibodies to herpes simplex virus type 1 proteins, including the immediate-early protein ICP 4. AB - Monoclonal antibodies were prepared against herpes simplex virus type 1 (strain 14012) by two immunization procedures. Procedure A utilized infectious virus propagated in mouse cells, and procedure B utilized mouse cells infected with herpes simplex virus in the presence of cycloheximide and harvested 1 h after removal of the inhibitor. A total of 52 monoclonal antibodies were obtained against 10 herpes simplex virus proteins, including four glycosylated proteins (a 110,000-molecular-weight protein, gB, gC, and gD) and six nonglycosylated proteins (a 68,000-molecular-weight protein, ICP 9, ICP 8, ICP 6, ICP 5, and the immediate-early ICP 4). The antibodies were assayed against herpes simplex virus types 1 and 2 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of radioimmunoprecipitates, immunofluorescence, and neutralization. Using the reagents prepared, we concluded that the 110,000-molecular-weight protein, gD, ICP 9, ICP 9, ICP 6, and the 68,000-molecular-weight protein express both type specific and cross-reactive antigenic determinants. In contrast, nine antibodies against gB all cross-reacted with herpes simplex virus type 2, whereas eight antibodies to gC all reacted type specifically. PMID- 6277789 TI - Pathogenesis of experimental skin infections induced by drug-resistant herpes simplex virus mutants. AB - The comparative analysis of the pathogenicity of a parental herpes simplex virus type 1 strain and its phosphonoacetic acid (PAA)-resistant and acyclovir (ACV) resistant mutants showed marked differences among them. After orofacial skin inoculation of hairless mice the parental and PAA-resistant viruses were detected during the first 4 days after infection at high and increasing titers in the trigeminal ganglia; the ACV-resistant mutant was present at low and decreasing titers in the ganglia. Severe and slow-healing skin lesions were produced by the parental and PAA-resistant viruses; mild and rapidly healing lesions were produced by the ACV-resistant mutant. Virus titers in ganglia and the intensity of skin lesions were related to the virus dose used in the primary infection. Latent infections became established in trigeminal ganglia of mice inoculated with 10(6.0) plaque-forming units of the parental or PAA-resistant virus; no latent infections were detected in ganglia of mice inoculated with 10(7.0) plaque forming units of the ACV-resistant mutant. Serum antibody titers attained similar values 4 weeks after primary infection with both mutants and the parental virus. Mice infected with the ACV-resistant mutant were reinfected with the parental and PAA-resistant viruses; the degree of protection against development of skin lesions, mortality, and latency was related to the dose of ACV-resistant virus used in the primary infection. Mortality was prevented by a dose of 10(6.0) plaque-forming units, skin lesions were prevented by a dose of 10(6.5) plaque forming units, and latency was prevented by a dose of 10(7.0) plaque-forming units of the ACV-resistant mutant. Protection against reinfection with the PAA resistant mutant was achieved with lower doses than protection against the parental virus. Serum antibody titers showed a 4- to 15-fold increase after reinfection. The results suggest that the ACV-resistant, latency-negative mutant has many attributes of a live attenuated herpes simplex virus vaccine. PMID- 6277790 TI - Cell-mediated immunity to herpes simplex virus: recognition of type-specific and type-common surface antigens by cytotoxic T cell populations. AB - In this communication, we examine the specificity of anti-herpes simplex virus (HSV) cytotoxic T lymphocytes (CTL). Serological studies of the two related HSV serotypes (HSV-1 and HSV-2) have revealed both type-specific and cross-reactive antigenic determinants in the viral envelope and on the surface of infected cells. By analysis of cytotoxicity of CTL, generated in vitro by restimulation of splenocytes from mice primed with one or the other HSV serotype, the recognition of both type-specific and cross-reactive determinants on infected target cells by anti-HSV CTL was detectable. Thus, effector cells generated by priming and restimulating with the same virus recognized both type-specific and cross reactive determinants on target cells infected with the homologous virus, but only cross-reactive determinants on target cells infected with the heterologous HSV serotype. CTL generated by restimulation with the heterologous virus were capable of recognizing only the cross-reactive determinants on either HSV-1- or HSV-2-infected target cells. These results indicate that two subpopulations of CTL exist in a population of anti-HSV immune spleen cells--those which recognize type-specific determinants and those specific for cross-reactive antigenic determinants present on the surface of HSV infected cells. The type-specific subset of anti-HSV CTL was shown to recognize the gC glycoprotein of HSV-1 infected target cells. In addition to the gC glycoprotein, at least one other type-specific surface antigen was also recognized by anti-HSV CTL in addition to the cross-reactive determinants recognized by anti-HSV CTL. PMID- 6277791 TI - Antibody responses in humans to individual proteins of herpes simplex viruses. AB - Sera from 231 women were used to examine their frequency of precipitation of various herpes simplex virus type 1 and 2 (HSV-1 and HSV-2) proteins and to determine if there was a rank order of immune responsiveness of humans to these HSV antigens. Radiolabeled viral proteins were reacted with serum and immune complexes isolated with staphylococcal protein A. Individual antigens were resolved by polyacrylamide gel electrophoresis and visualized by fluorography. As a group, these sera precipitated 31 HSV-1 and 27 HSV-2 proteins. HSV-1 polypeptides with molecular weights of 133,000, 99,000, and 82,000, as well as HSV-2 polypeptides with molecular weights of 131,000 and 101,000, were precipitated by essentially all sera that contained antibodies to HSV-1 and HSV 2. When attempts were made to order the viral proteins by constructing precipitation profiles ranking the antigens in patterns according to their frequency of precipitation, it was observed that the antigens were generally not ordered. Demographic analysis of the sera suggested that the differences in the number of proteins precipitated were associated with differences in age, education, age at first marriage, and income, which collectively may reflect the frequency of exposure to the virus. PMID- 6277792 TI - Protection against feline leukemia by vaccination with a subunit vaccine. AB - An effective vaccine against feline leukemia virus infection has been developed by the collection and concentration of tissue culture medium harvested from a tumor cell line. Lymphoid cells were grown to near saturation density in a normal growth medium and then transferred to a serum-free medium. The serum-free medium was collected, concentrated, and evaluated for its vaccine potential. Cats receiving the vaccine emulsified in complete Freund adjuvant developed high antiviral and antitumor titers and were protected (81%) against virus challenge. Cats receiving the vaccine without an adjuvant developed lower antibody levels and lower protection (53%) from viremia. Age-matched and litter-matched controls developed no antibody to test antigens before the challenge, and 100% became persistently viremic after the challenge. Vaccination with the soluble tumor cell antigen vaccine proved successful in preventing the induction of feline leukemia virus infection. PMID- 6277793 TI - Evidence for a bladder cell glycolipid receptor for Escherichia coli and the effect of neuraminic acid and colominic acid on adherence. AB - The rat bladder epithelial cell receptors involved in mannose-sensitive adherence of Escherichia coli strains were studied. Sodium metaperiodate and lipase pretreatment of epithelial cells significantly reduced bacterial adherence to cells whereas trypsin and phospholipase C had a marginal or insignificant effect on adherence. Neuraminidase and colominic acid significantly increased adherence, whereas N-acetylneuraminic acid significantly reduced adherence. These data suggest that the rat bladder epithelial cell receptors involved in mannose sensitive adherence are glycolipids. In addition, the data suggested that sialic acid on bladder epithelial cells acts as a nonspecific inhibitor of adherence, whereas colominic acid, a component of some E. coli K1 capsules, may act as a promoter of adherence. PMID- 6277795 TI - Inhibitory activity and bactericidal kinetics of mecillinam/ampicillin combinations against Enterobacteriaceae, Pseudomonas and Acinetobacter. AB - Mecillinam, ampicillin and combinations of the two were tested against Enterobacteriaceae by traditional agar dilution and under in vitro conditions simulating in vivo pharmacokinetics. Synergy was demonstrated by agar dilution in 30% of the strains. At simulated in vivo conditions, synergy between ampicillin and mecillinam was detected in a high osmolality medium, but not when the osmolality of the medium was low. The bactericidal effect was detected with mecillinam after one hour, while that of ampicillin or the combination was detected after 15 min. Changed bacterial morphology was observed by electron microscopy. Mecillinam produced ovoid spherical cell structures whereas combinations of the two drugs produced larger bacterial spheres. PMID- 6277794 TI - Murine cytomegalovirus stimulates natural killer cell function but kills genetically resistant mice treated with radioactive strontium. AB - Treatment of C3H/St mice with 100 microCi of 89Sr weakened their genetic resistance to murine cytomegalovirus (MCMV) infection. The criteria utilized to detect increased susceptibility were: (i) survival of mice; (ii) numbers of MCMV infected cells in the spleens and liver; and (iii) serum glutamic pyruvic transaminase levels. The natural killer (NK) cell activity of spleen cells from mice treated with 89Sr is very low. However, the NK activities of spleen cells of both normal and 89Sr-treated mice were greatly augmented 3 days after infection with MCMV. These NK cells lysed a variety of tumor cells and shared several features with conventional NK cells, but were not lysed by anti-Nk-1.2 serum (specific for NK cells) plus complement. Splenic adherent cells did not lyse tumor cells themselves but were necessary for the stimulation of NK cells by MCMV. The paradox of high NK cell function and poor survival in 89Sr-treated mice infected with MCMV was a surprise. We conclude that these augmented NK cells, of themselves, cannot account for the genetic resistance of C3H/St mice to infection with MCMV. PMID- 6277796 TI - Concentrations of cefotaxime and piperacillin in serum, subcutaneous tissue and muscle of patients undergoing dental surgery. PMID- 6277797 TI - Studies on long-term T-cell-mediated immunity to Epstein-BArr virus in immunosuppressed renal allograft recipients. AB - Peripheral blood lymphocytes from normal seropositive donors and renal transplant recipients on various immunosuppressive regimens have been tested for their ability to mount a cytotoxic response when cultured with autologous EB virus infected B cells and thereby to cause regression of proliferating b-cell foci. Cultures from 10 normal seropositive donors all showed the normal pattern of regression. Lymphocytes from patients receiving Cyclosporin A therapy with or without prednisolone completely failed to cause regression, thus allowing B-cell lines to proliferate unchecked. Cells from two of 17 patients treated with azathioprine and prednisone also failed to cause regression. The cells from the remaining 15 individuals showed regression response varying from minimal to normal. These results suggest a mechanism by which EB virus-related tumours may arise in immunosuppressed renal allograft recipients. PMID- 6277798 TI - Intestinal M3 antigen, a marker for the intestinal-type differentiation of gastric carcinomas. AB - The mucus-associated intestinal M3 antigen, normally restricted to intestinal goblet cells, was found in 35 out of 100 gastric adenocarcinomas belonging to intestinal (19/64) as well as diffuse (16/36) types according to Lauren's classification. often accompanying the other mucus-associated gastric M1 and M2 antigens. This M3 antigen was predominant over the gastric M antigens in 25 of these 35 tumors; 18 of these belonged to the histological intestinal type. According to the WHO classification, the M3 antigen was found to predominate in all mucinous adenocarcinomas (7/7), was never present in the undifferentiated carcinomas (0/8), but was also found in some tubulo-papillar (16/57) and signet ring cell (12/27) adenocarcinomas. This antigen could be used as a new criterion and incorporated into a point system containing morphological and tumor cell behavioral considerations; then it would appear to be a good marker for intestinal-type differentiation. Indeed, 22 of these 25 gastric adenocarcinomas which produced predominantly M3 antigen showed such an intestinal-like differentiation. The M antigen pattern of gastric carcinoma suggested a duodenal rather than colonic-type differentiation. PMID- 6277799 TI - The site of integration of the herpes simplex virus type 1 thymidine kinase gene in human cells transformed by an HSV-1 DNA fragment. AB - To analyze the site of integration of the herpes simplex virus type I (HSV-I) thymidine kinase (TK) gene in biochemically transformed human cells, TK-HeLa (BU25) cells were transformed to the TK+ phenotype by a cloned, 2 kbp Pvull fragment of HSV-I DNA. The transformed cells [HeLa(BU25)/TF pAGO PP3] were fused with mouse LM(TK-) cells, and human-mouse somatic cell hybrid clones (LH PP3 clones 1, 2, 3, 5 and 6) were isolated in HATG-ouabain selective medium. The HeLa(BU25)/TF pAGO PP3 cells and the LH PP3 hybrid clones expressed HSV-I specific TK activity and a herpesvirus-associated nuclear antigen, and contained herpesvirus nucleotide sequences. Molecular hybridization experiments were carried out to map the HSV-I and flanking cellular nucleotide sequences in the biochemically transformed cells. These experiments demonstrated that the HSV-I nucleotide sequences were integrated at a single site, and that the same cellular nucleotide sequences flanked the viral DNA in transformed HeLa(BU25)/TF pAGO PP3 and LH PP3 clone 5 cells. TK- revertant subclones isolated by growing the LH PP3 clone 5 cells in BrdUrd (and diphtheria toxin) failed to form colonies in HATG medium, but retained HSV-I nucleotide sequences. Isozyme analyses on 21 gene enzyme systems representing 21 human chromosomes revealed that all of the LH PP3 clonal lines expressed human hexosaminidase B, which has been assigned to chromosome 5, and all were sensitive to diphtheria toxin, which is also a marker for chromosome 5. Chromosome analyses showed that chromosome 5 was the nly human chromosome present in mitoses of LH PP3 clone 5 cells and that human chromosome 5 was present in most of the mitoses of LH PP3 clone 1, 2, 3, and 6 cells. The latter clones also contained 1 or 2 additional human chromosomes in some of the cells. As expected from the molecular hybridization analyses, TK- revertants of LH PP3 clone 5 cells retained portions of chromosome 5 and expressed human hexosaminidase B. The results indicate that HSV-I nucleotide sequences were stably integrated in the biochemically transformed cells, most likely in human chromosome 5. PMID- 6277800 TI - Heterotransplantation of small-cell carcinoma of the lung into nude mice: comparison of intracranial and subcutaneous routes. AB - We compared and contrasted intracranial (i.c.) and subcutaneous (s.c.) heterotransplantation of small-cell carcinoma of the lung (SCCL) into athymic nude mice. Fresh human SCCL tumor specimens, tumor colonies grown in soft agarose and continuous cell lines were used. Tumors induced by the three types of specimens were similar, but s.c. and i.c. transplants differed. S.c. tumors had longer latent times, were non-invasive and non-lethal. I.c. tumors has shorter latent periods, invariably grew in the meninges, frequently invaded and destroyed the underlying brain, and were lethal. The tumor-inducing dose for i.c. transplantation was 10 to 1,000 times lower than for s.c. transplantation. Pooled colonies of SCCL tumor specimens grown in soft agarose were inoculated i.c. While they contained relatively small numbers of cells (400-10,000), 83% of these colony specimens induced tumors after 58-243 days, confirming the "stem-cell" origin of the colonies. I.c. and s.c. transplants retained the characteristic morphology of SCCL, and, with one exception, did not metastasize to distant organs. Continuous cell lines could be established readily from both types of transplants, and they retained the characteristic cytology, growth and biochemical properties of the original SCCL tumors. I.c. heterotransplantation of SCCL is a useful tool, especially when small numbers of tumor cells are available, and may provide a model to study the biology and therapy of meningeal carcinomatosis. PMID- 6277801 TI - Altered properties of tumors induced by adenovirus type 12 DNA fragment transformed cells after growth in immunocompetent rats. AB - The GY1-3-1 cell line, which was derived from a rat cell line transformed by the Ad 12 HindIII-G fragment, can induce tumors in newborn rats but not in mature rats. However, when 10 mature rats were transplanted with a large number of GY1-3 1 tumor fragments, two developed tumors with a long latency period. These two tumors (GY1-3M and GY1-3Y) were quite different in properties from the originally inoculated GY1-3-1 tumor. Tumor transplantation studies revealed that both GY1-3M and GY1-3Y tumors could grow in mature rats and the number of takes with transplanted tumor was 100% with GY1-3M tumor and 40% with GY1-3Y tumor. Histopathologically original GY1-3-1 tumor showed a morphology of undifferentiated sarcoma, while the histologic picture of GY1-3M and GY1-3Y tumors was similar to that encountered in human fibrosarcoma and malignant fibrous histiocytoma, respectively. Both GY1-3M and GY1-3Y tumors, when recultured in vitro, gave rise to cell lines with a fibroblastic appearance although the original GY1-3-1 cell line exhibited an epithelioid morphology. In chromosome analysis, the GY1-3-1 cell line was pseudodiploid, while the cell lines from GY1-3M and GY1-3Y tumors were hyperdiploid or near-triploid. The metacentric marker chromosome, MI, was present in mitotic cells of the GY1-3-1 line and of the GY1-3M tumor lines, but absent from the GY1-3Y tumor lines. By Southern blot hybridization, multiple bands of cellular DNAs from parental GY1-3 1 cells hybridized with labelled Ad 12 HindIII-G, while only a single band hybridized DNAs from both GY1-3M and GY1-3Y tumors. PMID- 6277802 TI - Generation in vitro of HLA-restricted EB virus-specific cytotoxic human T cells by autologous lymphoblastoid cell lines: the role of previous EB virus infection and foetal calf serum. PMID- 6277803 TI - Characterization of human cells persistently infected with cytomegalovirus and exposed to a chemical carcinogen. AB - A non-productive, persistent human cytomegalovirus (CMV) infection was developed in human embryo lung (LU) cells by initially using supraoptimal temperatures to restrict the replication of a recent CMB isolate (76-24). Although CMV replication was observed in some cells within a few weeks of shifting the cultures to 37 degrees C and subculturing, CMV cytopathic effects and infectious CMV were not detected with further subculturing during the next 5 weeks. CMV specific nuclear antigens were, however, observed in most cells (designated LU-76 24) for 38 additional culture passages made during the following 12 months. Although the LU-76-24 cells were morphologically distinct from LU cell cultures carried in parallel as controls, the LU-76-24 were contact-inhibited in spite of a somewhat disoriented growth pattern. Treatment of LU-76-24 cells with 0.05 microgram/ml of the chemical carcinogen 4-nitroquinoline 1 oxide (NQO) induced a lytic CMV infection from which a clone of cells (designated LUC-NQO) was isolated. No effect was seen when LU cells were treated with the same dose of NQO. LUC-NQO cells were morphologically altered from LU-76-24 cells and had lost contact inhibition. Even though CMV-specific nuclear antigens were observed in LUC-NQO cells, infectious CMV was not detected in lysates of these cells, nor was CMV rescued by co-cultivation of these cells with susceptible cells or by induction with 5-iodo-2'-deoxyuridine. LUC-NQO, but not LU or LU-76-24 cells, plated with good efficiency in semi-solid medium. The results indicate that long term, non-productive, persistent CMV infections can be established in vitro and suggest that such cells may be more susceptible than non-infected cells to the action of chemical carcinogens. PMID- 6277804 TI - Establishment of B-lymphoblastoid cell lines from Marek's disease virus-induced tumors in turkeys. AB - Four lymphoblastoid cell lines were established from tumours of turkeys inoculated with high doses of the GA strain of Marek's disease virus (MDV). Unlike other MD lymphoblastoid cell lines of chicken origin, these MDV transformed turkey line appear to be B lymphocytes and produce immunoglobulin. Growth characteristics of these cell lines are slightly different; however, they all produce low levels of MDV-specific antigen and carry the complete genome of the virus as determined by virus rescue in the chicken or in duck embryo fibroblast cultures. These cell lines are pathogenic for chickens and produce virus-induced MD lesions. All four lines are free of the herpesvirus of turkeys, reticuloendotheliosis virus, and three lines are also free of avian leukosis virus. They all have typical normal turkey chromosomes and are positive for Marek's disease tumor-associated surface antigen. PMID- 6277805 TI - A ts T mutant of Schmidt Ruppin strain of Rous sarcoma virus restricted at 39.5 degrees C for the morphological transformation and the tumorigenicity of chicken embryo fibroblasts. AB - In order to investigate a possible correlation between in vitro transformation and tumorigenicity in ovo, a new temperature-sensitive class T mutant of Rous Sarcoma Virus was isolated with a lower (39 degrees 5C) restrictive temperature for morphological transformation. This lower restrictive temperature was compatible with the survival of chicken and duck eggs for the tumorigenicity studies. In chicken embryo fibroblasts (CEF) infected by this new mutant, PA 17, and cultured at 39 degrees 5C, increase of hexose uptake, plasminogen activator production and anchorage-independent growth were only partially restricted, requiring incubation at 41 degrees 5C for a complete shut-off. Tumorigenicity in chicken and duck eggs inoculated with CEF infected and transformed by PA 17 was restricted at 39 degrees 5C, correlating well with the restriction of morphological transformation at this temperature. The kinase activity of the transforming protein pp60src in lysates of PA 17 infected cells cultured at permissive or restrictive temperatures was labile in RIPA buffer, as in the case of some previously examined ts T mutants. In the non-ionic detergent NP40 buffer, the kinase activity of PA17 infected cell lysates was better conserved and showed a moderate temperature dependence. These results suggest that, in spite of the correlations between the transformed cell phenotype in vitro and cell tumorigenicity in ovo, it is difficult to establish a quantitative relationship. PMID- 6277807 TI - Analysis of human genital warts (condylomata acuminata) and other genital tumors for human papillomavirus type 6 DNA. AB - 32P-labelled cloned HPV 6 DNA was used as probe to analyze human genital tumors for DNA sequences homologous to HPV 6 DNA. Ninety three percent of all condylomata acuminata (41 out of 44) were found to harbor HPV 6 DNA. Of the remaining three, one contained HPV 1 DNA. No papillomavirus DNA was identified in the two other tumors. All three invasively growing giant condylomata acuminata (Buschke-Lowenstein tumors) investigated also contained HPV 6 DNA. Two out of six atypical condylomata of the cervix hybridized with HPV 6 DNA under stringent conditions, one only under conditions of low stringency. All DNA preparations from malignant tumors studies (54 cervical carcinomas, 10 penile carcinomas, two vulvar carcinomas) failed to anneal with HPV 6 DNA, even under conditions of low stringency. Although all HPV 6-positive condylomata acuminata analyzed in this study revealed HPV 6 DNA of regular molecular weight (5.1 x 10(6)), two of the Buschke-Lowenstein tumors, as well as one of the two positive atypical condylomata of the cervix, contained HPV 6 DNA with a remarkable size classes occurred in a supercoiled form without evidence for integration into host cell DNA. PMID- 6277806 TI - Detection of Epstein-Barr viral DNA internal repeats in the nasopharyngeal mucosa of Chinese with IgA/EBV-specific antibodies. AB - Fifty-six South Chinese individuals exhibiting IgA antibodies to EBV for 18 months and presenting nasopharyngeal abnormalities were biopsied. Four nasopharyngeal carcinomas, two at a very early stage, were detected. In 14 further individuals, without clinical or histopathological evidence of tumor, EBV/DNA internal repeats and/or EBNA were detected in the biopsied mucosae. The presence of IgA/EBV antibodies and/or EBV markers in the nasopharyngeal mucosa may characterize pre-cancerous conditions. PMID- 6277808 TI - The mechanism of genetic resistance to Marek's disease in chickens. AB - Genetic resistance to Marek's disease in RPL line-6 chickens is expressed not only at the level of host immunological responses against virus an tumour antigens, but also at the level of target lymphoid cells for virus infection and transformation. The nature of the target cell involved was investigated. Spleen cells from susceptible line-7 chickens adsorbed more Marek's disease virus and turkey herpesvirus in vitro than line-6 spleen cells. In the case of Marek's disease virus this was reflected in the replicative ability of the virus in vivo. Transplantation of thymus fragments from 1-day-old line-7 chickens into thymectomized line-6 chickens conferred a high degree of susceptibility on the latter, but the transplantation of spleen or fragments had no significant effect. The reverse procedure, i.e. grafting of line-6 thymi into line-7 chickens, did not diminish the susceptibility of the recipients. In each treatment group the observed titres of leukocyte-associated viraemia correlated with the susceptibility of the group to Marek's disease. Histologically the grafted thymus fragments became depleted of lymphocytes immediately after transplantation. By 6 days there was substantial recovery, apparently as a result of re-population of the thymic epithelium by host stem cells. This was confirmed by transplanting thymus fragments between individuals of opposite sexes. Karyotype analysis showed that the thymus contained lymphocytes of the sex of the recipient. However, karyotype analysis of lymphoma cells taken from recipient line-6 chickens that had received thymus grafts from line-7 birds of the opposite sex showed that, in the majority of cases, the lymphomas consisted of cells of donor origin. It is concluded that the susceptibility of line-7 chickens is largely attributable to the greater susceptibility of their T-lymphocytes to infection and transformation by Marek's disease virus, and that this susceptibility can be transferred to genetically resistant line-6 birds by adoptive transfer of the cells in the form of thymus fragments. PMID- 6277809 TI - Syngeneic and allogeneic cell-mediated cytotoxicity against Marek's disease lymphoblastoid tumor cell lines. AB - Cell-mediated cytotoxicity (CMC) of lymphocytes obtained from chickens infected with Marek's disease (MD) virus against allogeneic MD lymphoblastoid cell lines has been reported by several research groups. Recently, we established a number of cell lines from MD tumors obtained from highly inbred chickens and characterized for major and minor histocompatibility antigens. Allogeneic versus syngeneic CMC was studied using those cell lines and lymphocytes obtained from chickens 6-8 days post infection with SB-1, a non-oncogenic MD virus. Allogeneic cytotoxicity could be easily demonstrated, while syngeneic cytotoxicity was a rare event. However, increase of the CMC assay period from 4 to 8 h did enhance syngeneic cytotoxicity. Cold inhibition assays demonstrated that the allogeneic cytotoxicity was directed against alloantigens present on spleen lymphocytes sharing the same major histocompatibility antigens as the target cells. Cytotoxicity was not influenced by the sex of either target or effector cells or by the level of virus infectivity of the effector cells. PMID- 6277811 TI - Warfarin binding to plasma of workers exposed to toluene diisocyanate. AB - The extent of [14]C-warfarin binding to plasma proteins was evaluated in a group of normal, healthy volunteers and in two groups of individuals occupationally exposed to toluene diisocyanate (TDI). Plasma binding was assessed by ultrafiltration after the addition of racemic [14]C-warfarin to a final concentration of 0.8 microgram/ml. Chronic occupational exposure to TDI did not affect the extent of warfarin binding since warfarin free fractions (normalized to an albumin concentration of 4.5 g/dl) were 1.09 +/- 0.23 (mean +/- SD), 0.98 +/- 0.19, and 0.97 +/- 0.15 for controls and the two groups of TDI-exposed individuals, respectively. PMID- 6277810 TI - Kinetics of the immune response of tumor-bearing hamsters to two simian virus 40 coded non-structural polypeptides present in simian virus 40 tumor cells. AB - The kinetics of the immune response of hamsters transplanted with virus-free SV40 tumor cells (TSV5) to two SV40-coded non-structural proteins of 94,000 daltons (T antigen) and 17,000 daltons (t antigen) was studied in order to determine if the variation in the immune reactivity of serum from tumor-bearing animals towards these proteins can be explained on the basis of differential immune response during various stages of tumor growth. The results demonstrate that individual animals vary in their capacity to respond immunologically to T and t antigens in SV40-transformed cells and that, further, the immune response to T and t antigens is influenced by the stage of tumor development. PMID- 6277812 TI - The effects of exercise on plasma insulin. AB - The decrease of plasma insulin levels after physical training is explained mainly by a decreased insulin secretion, but also to a significant degree by an increased peripheral insulin metabolic clearance rate in several peripheral tissues and increased insulin uptake in the liver. The peripheral tissues demonstrate an increased insulin sensitivity, but this seems to be due to factors after the insulin receptor level. An increased turnover of insulin at the receptor level might be postulated. The explanation for the decrease of insulin secretion has been studied extensively. A decreased cortisol secretion might be a primary factor in physical training, followed by lower insulin levels. Autonomic nervous system adaptations could also be involved in the form of an increased alpha-adrenergic activity. Another alternative is that the effect is mediated via an influence of physical training on a gut "incretin" factor, facilitating insulin secretion, Finally, it might be considered that the increased insulin sensitivity of muscle following physical training is regulating insulin secretion via feedback mechanisms to the beta cell. Recent demonstrations of inhibited glucose uptake in the liver after exercise facilitates an explanation along these lines. PMID- 6277813 TI - The effectiveness of psychosocial support during post-surgical treatment of breast cancer. AB - Sixty-four women undergoing radiotherapy for breast cancer in a residential setting which offered structured cognitive and emotional support, are compared for level of psychological distress and satisfaction with staff support with 104 women living at home during treatment. The outcome measures were the 30-item Goldberg General Health Questionnaire (GHQ) and a 5-item author-constructed Treatment Support Index. A surprisingly large percentage of the sample did not, at any time, manifest appreciable psychological distress. For those women whose level of distress changed by a GHQ score of 3 or more over the three week course of treatment, residence in the Lodge was associated with decreased risk of change for the worse, and increased probability of change for the better. Results from the Treatment Support Index indicate that a significantly greater percentage of Lodge residents than non-residents were satisfied with the quantity, quality, and/or diversity of support available to them. PMID- 6277814 TI - Modification of radiation-induced division delay by caffeine analogues and dibutyryl cyclic AMP. AB - The mitotic selection procedure for cell cycle analysis was utilized to investigate the concentration-dependent modification of radiation-induced division delay in Chinese hamster ovary (CHO) cells by methyl xanthines (caffeine, theophylline, and theobromine) and by dibutyryl cyclic AMP. The methyl xanthines (concentrations from 0.5 to 1000 micrograms/ml) all reduced radiation induced division delay with the effect being linear between approximately 100 and 1000 micrograms/ml. After doses of 100-300 rad, delay was reduced by 75, 94 or 83 per cent at 1000 micrograms/ml for each drug, respectively. However, the addition of dibutyryl cyclic AMP had an opposite effect: radiation-induced delay was increased by the concentration range of 0.3 to 300 micrograms/ml. These results indicate that in mammalian cells the control of cell cycle progression and the modification of radiation-induced division delay are not simply related to intracellular levels of cyclic AMP. Rather, there appear to be at least two competing mechanisms which are differentially affected by caffeine analogues or by direct addition of dibutyryl cyclic AMP. The direct effect of caffeine and the methyl xanthines on membrane calcium permeability is considered. PMID- 6277815 TI - Immunization of rabbits with zeolite absorbed Trypanosoma gambiense inactive vaccine. AB - T. Gambiense was treated with natural zeolite, a very powdery mineral material which has strong absorptive capacity from the cationic exchangeable properties, and kept at 4 degrees C for 3 days. Rabbits injected with this inactivated vaccine were completely protected from a challenge inoculation of homologous viable parasites. Collected immune sera by weekly intervals from the immunized animals were tested for their ability to agglutinate trypanosomes. Serum antibodies were shown high titers 3 weeks post-inoculation, i.e., 1:512, and plateau of this titer was persistent up 3-4 weeks. For the experiment of passive protective effects of rabbit immune sera, mice were inoculated with 100 viable T. Gambiense by subcutaneous or intracerebral routes and then immediately performed two injection doses of 0.1 ml antiserum at intervals of 24 hours by subcutaneous. The protective ability was remarkable at the first to second weeks after the last immunization and then slightly decreased although a high level of agglutination titer remained in immune serum. PMID- 6277816 TI - Sweat gland tumors in the eyelids: a clinicopathological analysis of 55 cases. PMID- 6277817 TI - Tumors of the lacrimal sac: a clinicopathological analysis of 82 cases. PMID- 6277818 TI - Lacrimal gland tumors: a clinicopathological analysis of 160 cases. PMID- 6277819 TI - Human corneal stroma contains three distinct collagens. AB - Acetic acid-pepsin extracts of normal human corneal stromas were shown by carboxymethyl cellulose-column chromatography, sodium dodecyl sulfate-slab gel electrophoresis, highly purified collagenase, trypsin sensitivity, and cyanogen bromide peptide mapping to contain types I, III, and IV collagen, with type I by far the predominant species. This degree of collagen heterogeneity in normal human corneal stroma has not been reported previously and may be important in the understanding of wound healing and disease states. PMID- 6277820 TI - Mydriasis induced by tetrahydrocannabinol (THC) in rats. AB - Male albino rats, injected intravenously or intracerebroventricularly with delta 1-tetrahydrocannabinol (THC), develop mudriasis. The median effective dose of the intravenous administration group was 5 mg/kg THC, whereas that for the intracerebroventricular route was 150 microgram/kg THC. Sympathectomy significantly decreased the THC-induced mydriasis. The mydriatic effect was not influenced by naloxone. We conclude that THC produces mydriasis through a central action, the efferent pathway of which is the sympathetic system. PMID- 6277821 TI - Effect of delta 9-tetrahydrocannabinol on monoamine oxidase activity in bovine eye tissues, in vitro. AB - An investigation was undertaken to analyze the effects of delta 9 tetrahydrocannabinol (THC) on monoamine oxidase (MAO) activity from calf ocular tissues. Ciliary processes, retina, trabecular meshwork, choroid, and iris all demonstrated significant MAO activity in decreasing order of magnitude. THC in concentrations from 10(-8)M to 10(-12)M stimulated MAO activity in extracts from all five tissues, except for iris at 10(-8)M. Maximum stimulation of MAO activity occurred at the 1 x 10(-12)M level for all tissues. Retina, followed next by trabecular meshwork and then ciliary processes, exhibited the largest increase in MAO activity at 10(-12)M THC. This latter effect could possibly by related to the influence of THC on intraocular pressure. PMID- 6277822 TI - Cell surface glycoprotein patterns of two EBV-negative lines and their EBV converted sublines. AB - Cell surface galactosyl-glycoprotein patterns were compared between Ramos and BJAB, two Epstein-Barr virus (EBV)-negative Burkitt lymphoma lines, and their acutely EBV-superinfected and stably EBV-converted derivatives. A major difference between Ramos and its EBV-infected variants was the appearance of a 69,000 mol. wt. galactosyl-glycoprotein (GP69) in the latter. The same phenomenon was found in BJAB along with the additional appearance of a 71,000 mol. wt. galactosyl-glycoprotein (GP71) in the EBV-infected cells. GP71 was present in Ramos and in its EBV-converted derivatives. PMID- 6277823 TI - Cytomegalovirus replication in primary and passaged human placental cells. AB - Human cytomegalovirus (HCMV) was found to replicate in passaged fibroblastic human first trimester and term placental cells. The time-course of viral DNA replication as well as virus production in these human placental fibroblasts was similar to that in human embryo fibroblast cultures. In contrast, HCMV did not replicate in primary placental epithelioid cells. Continued passage (5 or more) of primary placental epithelioid cells was necessary to convert these cells to a state of permissiveness. The permissive cells were, however, fibroblasts. HCMV DNA replication in passaged placental fibroblastic cells was not affected by treatment with insulin or human chorionic gonadotropin. Furthermore, no replication of HCMV DNA occurred in choriocarcinoma cells, the epithelioid cells derived from cancer of the placenta. These results suggest that epithelial placental trophoblasts, either normal or transformed, were nonpermissive for HCMV. The permissiveness of HCMV infection to secondary placental cells which was observed might be due to the strong selection of fibroblastic cells in vitro. PMID- 6277824 TI - Linkage map of cotransfected viral DNA sequences in a mouse 3T3 cell line. AB - A physical map was constructed of a cotransformed 3T3 thymidine kinase (tk) negative cell line revealing a close linkage of the selectable herpes simplex virus type 2 tk gene and cotransfected simian virus 40 DNA sequences in high molecular-weight cellular DNA. PMID- 6277825 TI - Inhibition of the release of virion-associated murine leukemia virus genomic RNA by vesicular stomatitis virus. AB - The genomic RNAs of murine leukemia virus (MuLV) and vesicular stomatitis virus (VSV) were distinguishable based on a difference in sedimentation coefficient, polyadenylic acid content, and nucleotide sequence homology. Applying these biochemical characteristics, we found that the release of virion-associated MuLV genomic RNA from virus-producing cells was inhibited by VSV as early as 2 h after superinfection. This finding offers an explanation to our previous inability to detect MuLV(VSV) pseudotype formation during mixed infection with VSV and MuLV. PMID- 6277826 TI - Dicyclohexylcarbodiimide inhibition of succinate- and ubiquinol-cytochrome c reductase in beef heart mitochondria. AB - We have found that dicyclohexylcarbodiimide (DCCD) inhibits both the succinate cytochrome c and the ubiquinol-cytochrome c reductases in cytochrome c-depleted mitochondria. On the other hand the succinate-ubiquinone reductase is not decreased at the same levels of the inhibitor. The inhibition curve of DCCD results sigmoidal for succinate-cytochrome c reductase, whereas it is hyperbolic for the ubiquinol-1-cytochrome c reductase, with also a lower apparent KI. The inhibition appears dependent both on the time of preincubation and on the mitochondrial concentration. The apparent Km for ubiquinol-1 is increased and the maximal velocity of ubiquinol-cytochrome c reductase is decreased by DCCD. The effects do not appear to be caused by unspecific modification of the physicochemical state of the bc1 region of the respiratory chain. The results therefore suggest the presence of a DCCD-sensitive electron transfer step in the redox pathways from ubiquinol to cytochrome c. PMID- 6277827 TI - Neurological and electroencephalographic abnormalities after febrile seizures. Possible association with cytomegalovirus. AB - In order to assess the possible association of cytomegalovirus (CMV) with cerebral damage, 65 children aged between 2 and 10 years were tested for CMV isolation and antibody status. All of them had previously presented one or more episodes of febrile convulsions. At the time of our study, they were grouped according to their clinical features and electroencephalogram (EEG). 21 had typical and/or atypical absence attacks with 3 c/s spike-wave EEG (primary generalized epilepsy), 21 presented febrile seizures and spontaneous fits with focal EEG abnormalities, 23 had no seizures and a normal EEG. We performed the same investigation in 41 healthy children. The study showed a similar isolation rate of CMV in healthy subjects and in patients without neurological and EEG alterations, while a significantly higher isolation rate was observed in the groups with neurological and EEG abnormalities. As for serology, no significant difference was observed between the four groups, although the positivity rate of the healthy children was lower than in the other three groups. The study suggests a possible association of CMV with neurological and EEG abnormalities after febrile seizures. PMID- 6277828 TI - The Turcot syndrome. Case report. PMID- 6277829 TI - 5,8,11,14-Eicosatetraynoic acid (ETYA) inhibits binding of N-formyl-methionyl leucyl-phenylalanine (FLMP) to its receptor on human granulocytes. A note of caution. AB - Binding of two biochemically chemotactic factors, FMLP and C5a, to their respective receptors on human granulocytes produces a transient several-fold increase in cAMP. To determine if arachidonic acid metabolites were responsible for the increased cAMP concentrations, two inhibitors of arachidonic acid metabolism, indomethacin and ETYA, were incubated with granulocytes prior to FMLP or C5A. ETYA, but not indomethacin, inhibited both the cAMP and superoxide responses to a similar degree. However, the mechanism of this effect was found to be through inhibition of the binding of FMLP to its receptor rather than through inhibition of arachidonate metabolism. PMID- 6277830 TI - Vascular and neurologic complications following total hip replacement. PMID- 6277831 TI - Oxidative enzymes in the urinary apparatus of several marine fishes. AB - The distribution and activities of several oxidative enzymes in the urinary apparatus of seven marine fish species (hagfish, lesser spotted dogfish, electric ray, herring, marine catfish, cod, sea-horse) have been studied. Species were selected from three main taxonomic groups: Cyclostomata, Elasmobranchii and Teleostei. Distinctly positive enzyme reactions were found in the tubular elements of the kidney and the collecting duct-archinephric duct system, with the exception of the generally weak staining intensities for NADP-linked malate dehydrogenase. In the proximal tubule segment the second, ore distal part (PII) reacted, in general, very strongly when compared with the first proximal part (PI). If present, the distal tubule in teleosts showed only weak reactions, while this segment in elasmobranchs exhibited moderate to strong enzyme activities. In the epithelial cells of the collecting tubule-collecting duct system stronger reactions were confined to the glomerular teleost species, the corresponding part of the elasmobranch kidney showing weak staining intensities. In the urinary duct system distinctly positive enzyme reactions were only to be found in the archinephric duct of the teleost species, except for Plotosus. The ureters of the elasmobranchs exhibited weak enzymes activities throughout. The enzyme patterns of the various types of urinary tubules and ducts are compared with observations from several morphological and physiological studies. The histochemical findings are discussed in relation to corresponding investigations of freshwater fishes and problems arising from phylogenetic divergence of marine fish groups. PMID- 6277832 TI - Long term survivors with small cell carcinoma of the lung. PMID- 6277833 TI - Evaluation of misonidazole peripheral neurotoxicity in rats by analysis of nerve trains evoked response. AB - The clinical use of misonidazole and other nitroimidazole radiosensitizing agents is limited by the peripheral and central neurotoxicity that is produced in animals and humans. In a blinded study, rats treated with misonidazole at either 100 mg/kg or 300 mg/kg, 5 days/week for 3 weeks, were evaluated for peripheral neurotoxicity using nerve trains evoked responses. Only one rat treated at a dose of 100 mg/kg developed symptoms and signs of neurotoxicity, while all rats treated at 300 mg/kg developed these signs and symptoms. Nerve trains analysis made possible a diagnosis of neurotoxicity before overt clinical signs appeared. This test is non-invasive and may be useful for evaluating patients receiving nitroimidazole radiosensitizers as part of a radiation therapy regimen. PMID- 6277834 TI - Pulmonary gas exchange during altered density gas breathing. AB - The alveolar-arterial O2 partial pressure difference (PAO2 - PaO2) has been shown to decrease as carrier-gas density increases. This study was designed to confirm or deny the hypothesis that the improvement in O2 exchange is a result of density dependent changes in the alveolar ventilation-perfusion (VA/Q) distribution. On changing from heliox breathing to air breathing, there was an improvement in oxygen exchange along with a slight worsening of VA/Q distribution. The conclusion is reached that changes in VA/Q distribution due to altered carrier gas density are not responsible for changes in O2 exchange. A possible explanation is related to the interaction of diffusion and convection on inspiration, which may cause inspired gas distribution to be different from overall ventilation distribution. The interesting implication is that gas exchange properties of gases eliminated from the blood and exhaled are not necessarily symmetrical to the properties of gases inhaled and taken up by the blood. PMID- 6277835 TI - Renin, aldosterone, and converting enzyme during exercise and acute hypoxia in humans. AB - The possibility that hypoxia might inhibit the secretion of angiotension converting enzyme (ACE) would explain the low concentrations of aldosterone reported in humans at high altitude. To observe the effect of such a reduction in ACE concentration on the plasma aldosterone concentration (PAC) four subjects performed mild exercise throughout a 2-h study so as to elevate their plasma renin activity (PRA). After the first 60 min breathing air they were switched to breathing 12.8% O2 (4,000 an altitude equivalent). Venous samples were taken at intervals for hormone analysis. Results showed the expected rise of PRA and PAC both tending toward a plateau after about 45 min. There was no significant change in ACE activity (F = 0.065). Hypoxia produced a further 50% rise in PRA but a fall in PAC and a 30% reduction in ACE activity. Angiotensin I concentrations closely followed PRA throughout (r = 0.984). These results indicate that during exercise acute hypoxia changes the usual close relationship between PAC and PRA by reducing ACE activity. PMID- 6277836 TI - Ultrasound examination of pubertal girls and of patients with gonadal dysgenesis. AB - Girls with hypogonadism caused by hypothalamic lesions or chromosomal abnormalities need accurate pelvic exams to delinate pelvic structures. This work presents an evaluation of the usefulness of ultrasound examination of the pelvis in females with aberrant sexual development. Girls with these problems often need multiple pelvic examinations to monitor their uterine response to hormone therapy. Thirty-two patients with a variety of abnormalities including Mullerian agenesis and hypothalamic hypogonadism as well as patients with abnormal karyotype commonly associated with gonadal dysgenesis and male pseudohermaphrodism were studied. Repeat ultrasound scans were performed to quantitate uterine response to therapy. Ovarian visualization was usually possible, even in prepuberal girls. Following some patients at risk for gonadal tumors may be possible using repeat ultrasound examinations. The cooperation of the patients as well as of their families was gratifying, and the girls appeared to be more relaxed during ultrasound examination than during a pelvic examination. PMID- 6277837 TI - Plasma cortisol response to exogenous ACTH in 22 dogs with hyperadrenocorticism caused by adrenocortical neoplasia. AB - The plasma cortisol response to exogenous ACTH (ACTH stimulation test) was evaluated in 22 dogs with hyperadrenocorticism caused by adrenocortical neoplasia. The mean basal cortisol concentration (6.3 microgram/dl) was high, but 7 dogs had basal cortisol concentrations that were within normal range. Administration of exogenous ACTH increased the plasma cortisol concentrations in each dog. Normal post-ACTH cortisol concentrations were found in 9 (41%) of the 22 dogs; 13 (59%) had an exaggerated increase in cortisol concentrations after ACTH administration. In 9 of 13 dogs with carcinoma and in 4 of 9 with adenoma, the cortisol response was exaggerated. The mean post-ACTH cortisol concentration in the dogs with carcinoma was approximately 4 times that of the dogs with adenoma; the 7 dogs with the highest concentrations had carcinoma. Repeat studies were performed in 6 dogs 2 to 8 weeks after initial testing. In 5 of the 6 dogs, repeat testing yielded data of similar diagnostic significance. One dog, however, had an abnormally high post-ACTH cortisol concentration at initial evaluation, but had only a minimal response to ACTH administration, with a normal post-ACTH cortisol concentration, at time of resting. Although ACTH stimulation testing is useful in diagnosing hyperadrenocorticism, it can not reliably separate dogs with hyperfunction adrenocortical tumors from clinically normal dogs or from dogs with pituitary-dependent hyperadrenocorticism (bilateral adrenocortical hyperplasia). PMID- 6277838 TI - Studies of the deferent ducts from the testis of the African elephant, Loxodonta africana. III. Ultrastructure and cytochemistry of the ductuli efferentes. AB - The epithelium of the ductuli efferentes is composed of ciliated, principal, halo and basal cells. The supranuclear cytoplasm of ciliated cells is penetrated by particularly long cilial rootlets which are surrounded by numerous elongate mitochondria. Microtubules are arranged along the longitudinal axis of the cells. The spaces between the microvilli of principal cells form canaliculi which penetrate the apical cytoplasm and appear to be involved in endocytotic activity. The supranuclear cytoplasm contains oval mitochondria and numerous vacuoles. Both ciliated and principal cells contain poorly developed Golgi and endoplasmic reticulum, but numerous supranuclear dense bodies are usually present. Supranuclear and basal accumulations of dense bodies were identified as lipofuscin; they were the source of brown pigmentation in the proximal two thirds of the ductuli efferentes. The halo cells were probably macrophages. They occurred quite frequently and contained crescent shaped nuclei and large accumulations of lipofuscin material. PMID- 6277840 TI - A study of the relationship between dose and pharmacokinetics of ceftriaxone. PMID- 6277839 TI - Taste buds of the fungiform papillae in Cynomolgus monkey. AB - A study of the number of taste buds borne on 145 fungiform papillae from 20 Cynomolgus monkeys showed that, in contrast to the situation in man, most papillae are bud-bearing. The fine structure of taste buds on fungiform papillae of these monkeys was also examined. The cells contained clear apical vesicles, mitochondria, filament bundles and stacks of Golgi cisternae. Nerve fibres containing mitochondria, as well as clear and dense-cored vesicles, were scattered throughout the whole bud, but were more numerous near the base. It was, however, not possible to distinguish different cell types in these taste buds as a function of cell shape, electron density of the ground cytoplasm, presence of different organelles or relation to nerve processes. PMID- 6277841 TI - Journal of antimicrobial chemotherapy advice to contributors. PMID- 6277842 TI - Cefotaxime in serious infections--a clinical and pharmacokinetic study. PMID- 6277843 TI - The evolution of the broad-spectrum penicillins. PMID- 6277844 TI - In-vitro activity of Sch 29482, MK 0787, ceftriaxone and seven other antimicrobials against 840 separate clinical isolates. PMID- 6277845 TI - Carriers in electron transport from molecular hydrogen to oxygen in Rhizobium japonicum bacteroids. AB - An investigation has been conducted to identify electron transport carriers that participate in the oxidation of H2 by H2 uptake-positive strains of Rhizobium japonicum bacteroids. We have observed that the reduced form of dibromothymoquinone at a concentration of 0.2 mM strongly inhibited H2 uptake, endogenous respiration, and C2H2 reduction by bacteroid suspensions. Reduced dibromothymoquinone, however, failed to inhibit the transfer of electrons from H2 to methylene blue under anaerobic conditions, indicating that the hydrogenase per se is insensitive to this inhibitor. Metronidazole, at 1 mM, affected rates of H2 uptake and endogenous respiration only slightly, but strongly inhibited C2H2 reduction. Evidence for H2-dependent cytochrome reduction in an H2 uptake positive strain of R. japonicum bacteroids is presented. In kinetic studies, the rates of reduction of the type b and c cytochromes in the presence of H2 were shown to be severalfold higher than the rates due to endogenous respiration alone. With hydrogenase-deficient mutants of R. japonicum, no measurable effect of H2 on cytochrome reduction was observed. Our results indicate that ubiquinone and cytochromes of types b and c are involved in the oxyhydrogen reaction in R. japonicum. PMID- 6277846 TI - Respiration-dependent proton translocation in Nitrosomonas europaea and its apparent absence in Nitrobacter agilis during inorganic oxidations. AB - Oxygen pulse experiments were carried out with the nitrifying bacteria Nitrosomonas europaea and Nitrobacter agilis and with spheroplasts and everted vesicles prepared from Nitrobacter agilis. In addition to thiocyanate, the salting-in anions perchlorate and trichloroacetate proved to be permeant and effective in allowing respiration-dependent proton translocation with Nitrosomonas europaea. Valinomycin-K+, however, was generally ineffective in this respect with Nitrosomonas europaea. The observed leads to H+/O ratio for ammonium ion oxidation by Nitrosomonas europaea was 3.4; that for hydroxylamine and hydrazine cation oxidation was 4.4. These values, when corrected for production of stoichiometric protons and for the fact that the first step in ammonium ion oxidation (hydroxylamine production) is mediated by a monooxygenase, give effective leads to H+/O ratios of about 4 for these three substrates. This value compares favorably with those obtained with other aerobes. No convincing evidence was obtained for operation of a respiratory proton pump in Nitrobacter agilis during nitrite oxidation. Implications of this unexpected result are discussed. PMID- 6277847 TI - Effects of iron deficiency on heme biosynthesis in Rhizobium japonicum. AB - The effects of iron deficiency on heme biosynthesis in Rhizobium japonicum were examined. Iron-deficient cells had a decreased maximum cell yield and a decreased cytochrome content and excreted protoporphyrin into the growth medium. The activities of the first two enzymes of heme biosynthesis, delta-aminolevulinic acid synthase (EC 2.3.1.37) and delta-aminolevulinic acid dehydrase (EC 4.2.1.24), were diminished in iron-deficient cells, but were returned to normal levels upon addition of iron to the cultures. The addition of iron salts, iron chelators, hemin, or protoporphyrin to cell-free extracts did not affect the activity of these enzymes. The addition of levulinic acid to iron-deficient cultures blocked protoporphyrin excretion and also resulted in high delta aminolevulinic acid synthase and delta-aminolevulinic acid dehydrase activities. These results suggest the possibility that rhizobial heme biosynthesis in the legume root nodule may be affected by the release of iron from the host plant to the bacteroids. PMID- 6277849 TI - Naphthalene plasmids in pseudomonads. AB - A rapid method beginning with the direct lysis of bacteria in alkaline sodium dodecyl sulfate was used to detect naphthalene plasmids in pseudomonads. The strains NCIB 9816, PG, ATCC 17483, and ATCC 17484, which can grow on naphthalene as the sole source of carbon and energy, were examined. All except ATCC 17483 contained more than one plasmid. ATCC 17483 did not contain any plasmids. The largest pair of plasmids found in each of NCIB 9816 and PG(NAH2 and NAH3, respectively) determined naphthalene metabolism and could be transferred by conjugation. This also transferred the unusually regulated meta pathway enzymes for catechol metabolism. NAH2 determines the constitutive production of low concentrations of catechol 2,3-dioxygenase and 2-hydroxymuconic acid semialdehyde dehydrogenase, and NAH3 determines the constitutive production of high concentration of these. NAH2 and NAH3 gave identical fragments on digestion with BamHI or HindIII, but these were quite different from those of NAH. Nonetheless, NAH2 and NAH3 hybridized with NAH. PMID- 6277848 TI - Physical and genetic localization of ilv regulatory sites in lambda ilv bacteriophages. AB - A set of nine lambda dilv phages were used to transduce bacterial recipients containing point mutations or deletions in the ilv genes located at 84 min on the Escherichia coli K-12 chromosome. This genetic analysis indicated that two phages carry the entire ilvGEDAC cluster; others carry the complete ilvC gene and, in addition, bacterial DNA that extends to a termination point between ilvA and ilvC, within ilvD, within ilvE, or within ilvG. DNA extracted from the lambda dilv phages was digested with EcoRI, HindIII, KpnI, PstI, SalI, and SmaI. The restriction maps revealed that these phages were generated after insertion at four distinct insertion sites downstream (clockwise) of ilvC. The physical relationships between the various phages were further examined by electron microscopic heteroduplex analysis. The physical maps of the phages thus generated were straightforward and in complete accord with the genetic data. No evidence for genetic rearrangements of ilv DNA in the phage was obtained, thus validating conclusions based on the use of these phages in previous and ongoing research projects. Bacterial cells with deletions of the ilv genes were made lysogenic with lambda dilv phage to examine the regulation of ilv genes present in the phage. The results confirm previous studies showing that one site for control by repression and derepression is upstream (counterclockwise) of ilvG. It was shown, in addition, that the activities of dihydroxy acid dehydrase and threonine deaminase were increased when the prototrophic lysogens were grown with 20 mM leucine. Since this increase was exhibited even when the ilvG-linked control region was not carried by the lambda dilv phage, additional control sites must be located within the ilvEDA region of the ilvGEDA transcription unit. PMID- 6277851 TI - Partial nucleotide limitation induces phosphodiesterase I and 5'-nucleotidase in Bacillus subtilis. AB - Changes in the specific activity of enzymes involved in the degradation of RNA and nucleotides were measured in Bacillus subtilis under conditions of guanine deprivation, which initiates sporulation, and uracil deprivation, which does not initiate sporulation. Whereas the specific activities of most of the enzymes studied increased by less than a factor of 3, those of 5'-mononucleotide producing phosphodiesterase and 5'-nucleotidase increased at least eightfold under both deprivation conditions. PMID- 6277850 TI - MP13, a generalized transducing bacteriophage for Bacillus megaterium. AB - The first generalized transducing bacteriophage reported for Bacillus megaterium has been characterized. Optimum conditions for lysate production and transduction procedures were established so that transducing frequencies of 8 x 10(-6) and higher are now possible. The phage, MP13, has a head diameter of 97 nm and a contractile tail (202 by 17 nm) and adsorbs to the periphery of the cell. MP13 was inactivated rapidly at 60 degrees C, but not at 55 degrees C, and was sensitive to toluene, ether, and chloroform. When centrifuged in a neutral CsCl gradient, two bands were observed, a major band of 1.490 g cm-3 and a minor band of 1.482 g cm-3 buoyant density. The major band contained only infective particles, whereas the minor band contained both infective and transducing particles. Phage DNA was resistant to several restriction endonucleases, but yielded 9 fragments with MboI, more than 34 with HindIII, and 7 with BstEII. The molecular weights for the fragments from MboI-BstEII double digests total 97 x 10(9). PMID- 6277852 TI - Correlation between auxotrophy and plasmid alteration in mutant strains of Pseudomonas cepacia. AB - We describe here a class of mutants of Pseudomonas cepacia strain 249 in which different types of auxotrophy are associated with alterations in the 100 megadalton plasmid present in this strain. PMID- 6277853 TI - Cellular levels, excretion, and synthesis rates of cyclic AMP in Escherichia coli grown in continuous culture. AB - Changes in dilution rate did not elicit large and systematic changes in cellular cyclic AMP levels in Escherichia coli grown in a chemostat under carbon or phosphate limitation. However, the technical difficulties of measuring low levels of cellular cyclic AMP in the presence of a large background of extracellular cyclic AMP precluded firm conclusions in this point. The net rate of cyclic AMP synthesis increased exponentially with increasing dilution rate through either the entire range of dilution rates examined (phosphate limitation) or a substantial part of the range (lactose and glucose limitations). Thus, it is probable that growth rate regulates the synthesis of adenylate cyclase. The maximum rate of net cyclic AMP synthesis was greater under lactose than under glucose limitation, which is consistent with the notion that the uptake of phosphotransferase sugars is more inhibitory to adenylate cyclase than the uptake of other carbon substrates. Phosphate-limited cultures exhibited the lowest rate of net cyclic AMP synthesis, which could be due to the role of phosphorylated metabolites in the regulation of adenylate cyclase activity. Under all growth conditions examined, greater than 99.9% of the cyclic AMP synthesized was found in the culture medium. The function of this excretion, which consumed up to 9% of the total energy available to the cell and which evidently resulted from elaborate regulatory mechanisms, remains entirely unknown. PMID- 6277854 TI - Role of membrane-bound 5'-nucleotidase in nucleotide uptake by the moderate halophile Vibrio costicola. AB - Intact cells of Vibrio costicola hydrolyzed ATP, ADP, and AMP. The membrane-bound 5'-nucleotidase (C. Bengis-Garber and D. J. Kushner, J. Bacteriol. 146:24-32, 1981) was solely responsible for these activities, as shown by experiments with anti-5'-nucleotidase serum and with the ATP analog, adenosine 5'-(beta gamma imido)-diphosphate. Fresh cell suspensions rapidly accumulated 8-14C-labeled adenine 5'-nucleotides and adenosine. The uptake of ATP, ADP, and AMP (but not the adenosine uptake) was inhibited by adenosine 5'-(beta gamma-imido) diphosphate similarly to the inhibition of the 5'-nucleotidase. Furthermore, the uptake of nucleotides had Mg2+ requirements similar to those of the 5' nucleotidase. The uptake of ATP was competitively inhibited by unlabeled adenosine and vice versa; inhibition of the adenosine uptake by ATP occurred only in the presence of Mg2+. These experiments indicated that nucleotides were dephosphorylated to adenosine before uptake. The hydrolysis of [alpha-32P]ATP as well as the uptake of free adenosine followed Michaelis-Menten kinetics. The kinetics of uptake of ATP, ADP, and AMP also each appeared to be a saturable carrier-mediated transport. The kinetic properties of the uptake of ATP were compared with those of the ATP hydrolysis and the uptake of adenosine. It was concluded that the adenosine moiety of ATP was taken up via a specific adenosine transport system after dephosphorylation by the 5'-nucleotidase. PMID- 6277855 TI - Transformation of Bacillus stearothermophilus with plasmid DNA and characterization of shuttle vector plasmids between Bacillus stearothermophilus and Bacillus subtilis. AB - A thermophilic bacterium Bacillus stearothermophilus IFO 12550 (ATCC 12980) was transformed with each of the following plasmids, pUB110 (kanamycin resistance, Kmr), pTB19 (Kmr and tetracycline resistance [Tcr]), and its derivative pTB90 (Kmr Tcr), by the protoplast procedure in the presence of polyethylene glycol at 48 degrees C. The transformation frequencies per regenerant for pUB110, pTB19, and pTB90 were 5.9 x 10(-3), 5.5 x 10(-3), and 2.0 x 10(-1), respectively. Among these plasmids, pTB90 was newly derived, and the restriction endonuclease cleavage map was constructed. When tetracycline (5 micrograms/ml) was added into the culture medium, the copy number of pTB90 in B. stearothermophilus was about fourfold higher than that when kanamycin (5 micrograms/ml) was added instead of tetracycline. Bacillus subtilis could also be transformed with the plasmids extracted from B. stearothermophilus and vice versa. Accordingly, pUB110, pTB19, and pTB90 served as shuttle vectors between B. stearothermophilus and B. subtilis. The requirements for replication of pTB19 in B. subtilis and B. stearothermophilus appear to be different, because some deletion plasmids (pTB51, pTB52, and pTB53) derived from pTB19 could replicate only in B. subtilis, whereas another deletion plasmid pTB92 could replicate solely in B. stearothermophilus. Plasmids pTB19 and pTB90 could be maintained and expressed in B. stearothermophilus up to 65 degrees C, whereas the expression of pUB110 in the same strain was up to 55 degrees C. PMID- 6277856 TI - Identification of the structural genes for glutamate synthase and genetic characterization of this region of the Salmonella typhimurium chromosome. AB - Salmonella typhimurium cells require glutamate synthase activity for growth in media containing a growth rate-limiting nitrogen source. Although this enzyme plays a critical role in ammonia assimilation, little is known about the organization and regulation of the structural genes for its two subunits. To identify the location of the structural genes, mutants having heat-labile glutamate synthase activities were isolated and characterized. Mutations that altered glutamate synthase activity were mapped at 69 U on the S. typhimurium chromosome. Four strains with independent Tn10 insertions in this region were constructed and used for mutant selection and for positioning mutations affecting glutamate synthase activity relative to other genetic markers. In contrast to results obtained with Escherichia coli mutants, there was no linkage between mutations affecting glutamate synthase activity and the argG gene. The results of a combination of transduction experiments demonstrated the gene order argG-glnF gltB-cod-argR-envB-aroE for S-typhimurium. PMID- 6277858 TI - Transfer of chimeric plasmids among Salmonella typhimurium strains by P22 transduction. AB - Salmonella typhimurium bacteriophage P22 transduced plasmids having P22 sequences inserted in the vector pBR322 with high frequency. Analysis of the structure of the transducing particle DNA and the transduced plasmids indicates that this plasmid transduction involves two homologous recombination events. In the donor cell, a single recombination between the phage and the homologous sequences on the plasmid inserted the plasmid into the phage chromosome, which was then packaged by headfuls into P22 particles. The transducing particle DNA contained duplications of the region of homology flanking the integrated plasmid vector sequences and lacked some phage genes. When these defective phage genomes containing the inserted plasmid infected a recipient cell, recombination between the duplicated regions regenerated the plasmid. A useful consequence of this sequence of events was that genetic markers in the region of homology were readily transferred from phage to plasmid. Plasmid transduction required homology between the phage and the plasmid, but did not depend on the presence of any specific P22 sequence in the plasmid. When the infecting P22 carried a DNA sequence homologous to the ampicillin resistance region of pBR322, the vector plasmid having no P22 insert could be transduced. P22-mediated transduction is a useful way to transfer chimeric plasmids, since most S. typhimurium strains are poorly transformed by plasmid DNA. PMID- 6277857 TI - Identification of citrate utilization transposon Tn3411 from a naturally occurring citrate utilization plasmid. AB - We have isolated a new transposon, Tn3411, encoding citrate-utilizing ability, from a naturally occurring citrate utilization (Cit) plasmid, pOH3001. Citrate transposon Tn3411 was transposed from pOH3001 to lambda b519 b515 cI857 S7 (abbreviated lambda bb) phage, and further from the resulting lambda bb:Tn3411 to a vector plasmid, pBR322, in recA-deficient strains. The Cit+ plasmids (pOH2 and pOH3) constructed by the integration of Tn3411 into pBR322 were examined by restriction endonuclease and heteroduplex analysis. The results obtained were as follows: (i) Tn3411 was 7.4 kilobases long and flanked by small inverted repeats, and it contained one more pair of inverted repeats at the opposite orientation in the internal region, thus making alternate repeats; and (ii) the Cit+ structure gene was located on the fragment (5.5 kilobases) between two SalI cleavage sites on Tn3411. PMID- 6277859 TI - Recombination between two IS/s flanking the r-determinant of R100-1: involvement of dor and recA gene functions in Salmonella typhimurium. AB - Drug resistance genes of the r-determinant component of a composite R plasmid R100-1 were frequently lost in Salmonella typhimurium. Various deletion mutants were analyzed by restriction endonuclease cleavage, Southern blotting, and hybridization techniques. The loss of the r-determinant was found to be the result of a reciprocal recombination between the two IS/s flanking the r determinant. This recombination depended upon both dor and recA gene functions. PMID- 6277860 TI - Utilization of a thermosensitive episome bearing transposon TN10 to isolate Hfr donor strains of Erwinia carotovora subsp. chrysanthemi. AB - A thermosensitive episome bearing the transposon Tn10, F(Ts)::Tn10 Lac+, has been successfully transferred from Escherichia coli to several wild strains of the enterobacteria Erwinia carotovora subsp. chrysanthemi, which are pathogenic on Saintpaulia ionantha. In one of these strains, all of the characters controlled by this episome (Lac+, Tetr, Tra+) were expressed, and its replication was stopped at 40 degrees C and above. At 30 degrees C, the episome was easily transferred between strains derived from E. carotovora subsp. chrysanthemi 3937j and to E coli. Hfr donor strains were obtained from a F' strain of 3937j by selecting clones which grew at 40 degrees C on plates containing tetracycline. One of these strains, Hfrq, was examined in more detail: the characters Lac+ and Tetr were stabilized and did not segregate higher than its parental F' strain. The mating was most efficient at 37 degrees C on a membrane. Hfrq transferred its chromosome to recipient strains at high frequency and in a polarized fashion, as evidenced by the gradient of transfer frequencies, the kinetics of marker entry (in interrupted mating experiments), and the analysis of linkage between different markers. The chromosome of Hfrq was most probably transferred in the following sequence: origin...met...xyl...arg...ile...leu...thr...cys...pan...ura...gal...trp...his. ..pur... Moreover, this genetic transfer system proved to be efficient in strain construction. PMID- 6277861 TI - Rhizobium japonicum mutants that are hypersensitive to repression of H2 uptake by oxygen. AB - The synthesis of an H2 oxidation system in free-living Rhizobium japonicum wild type strain SR is repressed by oxygen. Maximal H2 uptake rates were obtained in strain SR after derepression in 11 microM or less dissolved oxygen. Oxygen levels above 45 microM completely repressed H2 uptake in strain SR. Five R. japonicum mutant strains that are hypersensitive to repression or H2 oxidation by oxygen were derived from strain SR. The mutants were obtained by screening H2 uptake negative mutants that retained the ability to oxidize H2 as bacteroids from soybean nodules. As bacteroids, the five mutant strains were capable of H2 oxidation rates comparable to that of the wild type. The mutants did not take up H2 when derepressed in 22 microM dissolved oxygen, whereas strain SR had substantial activity at this oxygen concentration. The O2 repression of H2 uptake in both the wild-type and two mutant strains, SR174 and SR200, was rapid and was similar to the effect of inhibiting synthesis of H2 uptake system components with rifampin. None of the mutant strains was able to oxidize H2 when the artificial electron acceptors methylene blue or phenazine methosulfate were provided. The mutant strains were not sensitive to killing by oxygen, they took up O2 at rates similar to strain SR, and they did not produce an H2 uptake system that was oxygen labile. Cyclic AMP levels were comparable in strain SR and the five mutant strains after subjection of the cultures to the derepression conditions. PMID- 6277862 TI - Direct DNA repeat in plasmid R68.45 is associated with deletion formation and concomitant loss of chromosome mobilization ability. AB - Plasmid R68.45 has been useful in promoting the transfer of chromosomal markers in bacteria of many genera. In donors harboring R68.45, chromosome-mobilizing ability (Cma) may be lost without the loss of other plasmid markers. However, Cma can be somewhat stabilized by maintenance of the donors in the presence of kanamycin (Km). We isolated variants of R68.45 from four bacterial species of three genera. Plasmid variants isolated included those without Cma, without transfer function (Tra) and Cma, or without Tra, Cma, and Km resistance. In Erwinia carotovora subsp. atroceptica EA153, the loss of plasmid markers is dependent on the culture medium on which the donors are maintained. Restriction endonuclease analyses of the variant plasmids revealed that most are deletion mutants of R68.45. In all cases when the uncertainty in the ends of the deletions was not too great, one end of the deletion was shown to originate within or near the direct DNA duplication in R68.45 which is required for Cma and which maps close to the Km resistance determinant. Furthermore, the types of deletions observed are consistent with what might be expected for deletions generated by tandemly repeated insertion sequences. Therefore, we suggest that the DNA duplication is the source of much of the instability observed in R68.45. However, data are presented for E. carotovora subsp. atroceptica EA153 which suggest that another region of R68.45 may also play a role in its stability in this species. PMID- 6277863 TI - Comparison of transcription of beta-lactamase genes specified by various ampicillin transposons. AB - The beta-lactamase gene from four kinds of ampicillin transposons, Tn2601, Tn3, Tn2602 and Tn1, specifying the type I (or TEM type, alternatively) beta-lactamase was cloned onto plasmid pACYC184, and the level of in vivo transcription from each beta-lactamase gene was determined by DNA-RNA hybridization. Type I beta lactamase is very uniform enzymologically, but heterogeneous in absolute levels of enzyme activity. The results demonstrated that the heterogeneity can be explained by the efficiency of transcription of each beta-lactamase gene, suggesting a difference in its promoter efficiency. A comparison of the levels of transcription of the beta-lactamase gene and the whole ampicillin transposon suggested that the beta-lactamase gene has the strongest promoter all of the genes in the ampicillin transposon. PMID- 6277864 TI - Isolation of a trisaccharide containing 2-amino-2-deoxy-D-galacturonic acid from the Bordetella pertussis endotoxin. AB - 4-O-(2-Amino-2-deoxy-alpha-D-glucopyranosyl-6-O-(2-amino-2-deoxy-alpha-D galactopyranuronyl)-D-glucopyranose, a branched-chain trisaccharide, was isolated after hydrolysis of Bordetella pertussis endotoxin with 4 M HCl for 1 h at 100 degrees C. The trisaccharide was present in both polysaccharide moieties of the two constituent lipopolysaccharides of this endotoxin. Its structure was established by analysis of the 400-MHz nuclear magnetic resonance spectrum and by chemical and enzymatic degradation. PMID- 6277865 TI - Cyclic AMP may not be involved in catabolite repression in Saccharomyes cerevisiae: evidence from mutants capable of utilizing it as an adenine source. AB - Mutants able to utilize 5'-AMP or cyclic AMP as the adenine source were isolated from an ade6 ade10 double mutant by ethyl methane sulfonate mutagenesis. A single amp1 mutation, primarily selected on 5'-AMP medium, confers the phenotype for utilization of exogenous 5'-AMP as the adenine source. From the ade6 ade10 amp1 triple mutant, a mutant able to utilize cyclic AMP was isolated, and the mutant phenotype was proven to be due to the simultaneous occurrence of triple mutations designated as cam1, cam2, and cam3. The cam3 mutation, but not cam1 or cam2, also confers the phenotype for utilizing 5'-AMP, the same phenotype as the amp1 mutation. All of these mutations are recessive to the respective wild-type counterparts. Cells having the ade6 ade10 amp1 cam1 cam2 cam3 genotype showed significant ability to take up exogenous cyclic AMP, whereas no differences were observed in cyclic AMP phosphodiesterase activity in comparison with that of the original strains used in the mutant isolation. Since glucose severely repressed galactokinase synthesis in the constitutive GAL81 mutant having the ade6 ade10 amp1 cam1 cam2 cam3 genotype, irrespective of the presence or absence of cyclic AMP in the medium, it was suggested that cyclic AMP is not involved in the mechanism of catabolite repression in Saccharomyces cerevisiae. It does, however, have a stimulative effect on the galactokinase synthesis in the GAL81 mutant in the absence of glucose. PMID- 6277866 TI - dnaC-dependent reconstitution of replication forks in Escherichia coli lysates. AB - Lysates of Escherichia coli exhibit a DNA-synthesizing activity that depends on the presence of replication forks and of replication proteins. Replicative activity was reconstituted in vitro by mixing lysates prepared from temperature sensitive dnaB mutants with wild-type dnaB protein. Lysates of double mutants deficient in both dnaB and dnaC genes could only be complemented by the addition of both dnaB and dnaC proteins, whereas lysates deficient in dnaC protein did not require the addition of any exogenous factor. This shows that the replication machinery, once it is running along the chromosome, is independent of dnaC protein, dnaC activity, however, is required for the replacement of defective dnaB protein at running replication forks. PMID- 6277867 TI - Respiratory physiology and energy conservation efficiency of Campylobacter jejuni. AB - A study of the electron transport chain of the human intestinal pathogen Campylobacter jejuni revealed a rich complement of b- and c-type cytochromes. Two c-type cytochromes were partially purified: one, possibly an oxidase, bound carbon monoxide whereas the other, of high potential was unreactive with carbon monoxide. Respiratory activities determined with membrane vesicles were 50- to 100-fold higher with formate and hydrogen than with succinate, lactate, malate, or NADH as substrates. Evidence for three terminal respiratory components was obtained from respiratory kinetic studies employing cyanide, and the following Ki values for cyanide were determined from Dixon plots: ascorbate + reduced N,N,N', N'-tetramethyl-p-phenylenediamine, K1 + 3.5 muM; malate, K1 = 55 muM; and hydrogen, K1 = 4.5 muM. Two oxidases (K1 = 90 muM, 4.5 mM) participated in the oxidation of succinate, lactate, and formate. Except with formate, 37 muM HQNO inhibited respiration by approximately 50%. Carbon monoxide had little inhibitory effect on respiration except under low oxygen tension (less than 10% air saturation). The stoichiometry of respiratory-driven proton translocation (H+/O) determined with whole cells was approximately 2 for all substrates examined except hydrogen (H+/) = 3.7) and formate (H+/O = 2.5). The higher stoichiometries observed with hydrogen and formate are consistent with their respective dehydrogenase being located on the periplasmic face of the cytoplasmic membrane. The results of this study suggest that the oxidation of hydrogen and formate probably serves as the major sources of energy for growth. PMID- 6277868 TI - Isoelectric focusing and crossed immunoelectrophoresis of heme proteins in the Escherichia coli cytoplasmic membrane. AB - Isoelectric focusing (IEF), agarose electrophoresis, and crossed immunoelectrophoresis (CIE) were used to resolve the heme-containing proteins of the Escherichia coli cytoplasmic membrane after solubilization by Triton X-100. Two bands in IEF stained for heme with pI values of 4.7 and 5.3. One of the bands, with an isoelectric point of pH 5.3, was present only when the cells were grown to late log or stationary phase and possessed N,N,N,'N'-tetramethyl-p phenylene-diamine (TMPD) oxidase activity. The pI 4.7 band was present in cells harvested in both mid-log and stationary phases. Agarose electrophoresis, using larger samples, revealed the same two components apparent by IEF, and, in addition, a third component. The heme-containing fractions were extracted after agarose electrophoresis and subjected to further study. The component which was present in cells grown to stationary phase contained hemes b, a1, and d. The other two fractions contained only b heme. One of these corresponded to the component with pI 4.7 in IEF and had catalase activity. Antisera were raised against Triton X-100-solubilized cytoplasmic membranes and against the focused TMPD oxidase complex. With these anti-sera, CIE in the presence of Triton X-100 revealed four precipitin complexes containing heme. Three of these corresponded to the components identified by IEF and agarose electrophoresis. We demonstrate that the combined use of IEF and CIE is valuable for analysis of membrane proteins. In particular, this work represents a substantial initial step toward a structural elucidation of the E. coli aerobic respiratory chain. PMID- 6277869 TI - DNA homology of the promoter-distal regions of the tra operons of sex factors F and R100 in Escherichia coli K-12. AB - The promoter-distal regions of the tra operons of F and R100-1 were analyzed by heteroduplex analysis, and the regions of nonhomology were identified. A common EcoRI restriction site was shown to be present, and this has allowed the physical maps to be aligned. PMID- 6277870 TI - Construction of a hybrid plasmid capable of replication in the bacterium Escherichia coli and the cyanobacterium Anacystis nidulans. AB - A hybrid plasmid was constructed between the 5.3-megadalton plasmid (pUH24) of Anacystis nidulans R2 and the Escherichia coli plasmid pBR322. This was accomplished by adding a transposon to pBR322 and transforming this DNA into A. nidulans. One resultant hybrid, pLS103, had a molecular weight of 6.8 x 10(6), replicated in both organisms, had unique sites for two restriction endonucleases, conferred ampicillin resistance on both organisms, and could be used as a cloning vector in A. nidulans. PMID- 6277871 TI - Determining the phoM map location in Escherichia coli K-12 by using a nearby transposon Tn10 insertion. AB - A phoR strain was constructed with transposon Tn10 inserted near the phoM+ locus. This was done without any prior knowledge of the phoM map location. Subsequently, we defined the phoM map position by screening tetracycline-sensitive (Tcs) derivatives for mutants which were both alkaline phosphatase negative (ther phoR phoM double mutant phenotype) and auxotrophic simultaneously. Some of these mutants were Thr-. Bacteriophage P1-mediated transductions were used to confirm that phoM and its nearby Tn10 insertion were closely linked to thr. Unexpectedly, 7 of 10 mutants analyzed also had mutations unlinked to the phoM-thr-Tn10 region. These may represent a new type of Tn10-promoted molecular event which is caused by transposition of a Tn10 end (IS10). PMID- 6277872 TI - Development of broad-host-range vectors and gene banks: self-cloning of the Pseudomonas aeruginosa PAO chromosome. AB - A host-vector system for Pseudomonas aeruginosa PAO was developed. Scattered regions of the strain PAO chromosome were cloned by direct selection for complementation of auxotrophs or from a DNA gene bank which contains over 1,000 independently isolated chromosome-vector recombinant plasmids. The use of partially digested chromosomal DNA facilitated the selection of a variety of strain PAO chromosomal markers. The progenitor of the vector was a small, multicopy plasmid, pRO1600, found in a PAO strain which had acquired RP1 in a mating experiment. The bacterial host range that could be determined by transformation of vectors produced from pRO1600 resembles that for plasmid RP1. Two derivative plasmids were formed: pRO1613, for cloning DNA cleaved with restriction endonuclease PstI, and pRO1614, which was formed by deleting part of pRO1613 and fusion with plasmid pBR322. Plasmid pRO1614 utilizes known cloning sites within the tetracycline resistance region of pBR322. PMID- 6277873 TI - Analysis of the promoter-distal region of the tra operon of the F sex factor of Escherichia coli K-12 encoded by EcoRI restriction fragments f17, f19, and f2. AB - The promoter-distal region of the tra operon of the F sex factor Escherichia coli K-12 was analyzed, using the chimeric plasmid pRS31, which contains the F EcoRI restriction fragments f17, f19, and f2 cloned into the EcoRI site of pSC101. A series of deletion plasmids of pRS31, extending increasing distances from a site in f17 through f19 and ending in f2, were isolated. These plasmids were examined by heteroduplex analysis with the parent DNA, and a restriction map of this region of DNA was constructed. A series of Tn5 insertion derivatives of pRS31 were also isolated and mapped, using both heteroduplex analysis and restriction mapping. Both the insertion and deletion mutants were tested in minicells for the synthesis of radioactively labeled proteins. This allowed the identification of the individual gene products and mapping of the genes. The result is a saturated physical map of this region of DNA from fragment f17 through to the IS3 insertion sequence near the promoter-distal end of f2. PMID- 6277875 TI - Purification and characterization of an inhibitor of calcium-activated neutral protease from rabbit skeletal muscle. AB - An endogenous inhibitor of calcium-activated neutral protease was purified to homogeneity from rabbit skeletal muscle using ion-exchange chromatography on DEAE cellulose and QAE-Sephadex A-50 columns, chromatofocusing, and hydrophobic interaction chromatography on a phenyl-Sepharose CL-4B column. The purified inhibitor was shown to be a dimer of identical subunits and each subunit has a molecular weight of about 34,000. This inhibitor was remarkably thermo- and acid stable. It was specific for calcium-activated neutral protease and had no effect on any other protease examined (trypsin, papain, alpha-chymotrypsin, bromelain, etc.). It is demonstrated that the inhibition is due to the formation of stoichiometric complex between two enzyme molecules and one inhibitor molecule. PMID- 6277874 TI - Promoter-distal region of the tra operon of F-like sex factor R100 in Escherichia coli K-12. AB - The distal region of the tra (transfer) operon of F-like plasmid R100 was investigated, using small plasmids derived from R100, primarily the plasmid pSM6. The transposon Tn5 (which confers kanamycin resistance) was inserted at different positions into pSM6, and the transposition derivatives were tested for ability to complement defined tra mutants of the F sex factor. Thus, the tra genes traH, G, T, and D were localized on the plasmid R100. A restriction map of pSM6 was constructed, and the locations of the insertions were mapped, using restriction endonuclease digestion of the plasmid DNA and exploiting the fact that several restriction sites are localized in the inverted repeat regions of the transposon. The gene products of the genes traG, S, T, and D were identified by radioactive labeling of proteins synthesized in minicells carrying the various insertion plasmids followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The presence of another transfer gene, traI, was inferred from these data. Another protein, the r2-A protein, was also identified, and its gene was mapped. On the basis of the data, a best-fit physical map of this region of the tra operon of R100 was constructed. The results confirmed that the general order and size of the distal transfer genes is as in the F sex factor, but showed that differences exist with respect to all of the gene products. The significance of these differences are discussed in the light of the genetic and physical homology (Manning et al., J. Bacteriol. 150:76-88) of the transfer regions. PMID- 6277876 TI - Substrate-concentration of dependences of tension, shortening velocity and ATPase activity of glycerinated single muscle fibers. AB - Tension P0 and ATPase activity J0 of glycerinated single muscle fibers under isometric concentration as well as the velocity V0 of unloaded shortening were measured as a function of substrate concentration [S]. The stiffness of fibers with sinusoidal length changes at 1 kHz was used as a qualitative measure of the amount of rigor complex. P0 is an increasing function of [S] at low substrate concentrations and has a broad maximum at about 10-40 micrometers MgATP. In this concentration range, 10-40 micrometers, V0 still has a very small value. Then it increases and finally reaches at a plateau at about 1 mM MgATP. J0 increases as P0 does. However, it reaches at a saturated level at about the same concentration as V0. Either 0.5 mM 8-BrATP or 1 mM PPi was added to the substrate solutions to reduce the amount of rigor complex at low substrate concentrations. The addition of PPi of 8-BrATP decreases P0 dominantly at low concentrations of substrate and shifts the maximum to about 100 micrometers MgATP. 8-BrATP considerably increases V0 at low substrate concentrations while V0 is decreased by added PPi. The temperature coefficients, Q10 values were obtained for P0, J0, and V0. The values are essentially constant, 2.1-2.4, in the cases of P0 and J0, and about the same values were found for V0 at very low substrate concentrations. However, they become about 3.3 in the concentration range from 34 micrometers and 2.3 mM. The P V relation was obtained at 11 micrometers and 2.3 micrometers MgATP. The normalized P-V relation at 11 micrometers was unchanged when 8-BrATP was added. The results are discussed in connection with the mechanism of actomyosin-ATPase activity as well as that of the elementary cycle of the motive force generation. PMID- 6277877 TI - Site-specific endo-deoxyribonucleases in eukaryotes: endonucleases of yeasts, Saccharomyces and Pichia. PMID- 6277878 TI - On cytochrome c3 folding. AB - The structure of cytochrome c3 from Desulfovibrio vulgaris Miyazaki (DvM) is considered in detail by scrutinizing main chain folding together with the structure of the protein from D. desulfuricans Norway (DdN). The relative arrangement of the four heme groups in this molecule is similar to that of DdN and the disposition of alpha-carbon atoms of cysteine and histidine residues binding to heme groups is also similar. Structural differences between the two proteins occur in the shape of some specific loops of the chain on the molecular surface. As a result of a careful comparison of structures and sequences in both cytochromes c3, the reported sequence alignment of the cytochrome c3 family has been revised. Our new proposal of a sequence alignment based on the three dimensional structures contains 24 evolutionarily conservative residues. All these conservative residues may play an important role in the folding pattern of cytochromes c3. PMID- 6277879 TI - Autolysis of calcium-activated neutral protease of chicken skeletal muscle. AB - The conditions and process of autolysis of calcium-activated neutral protease (CANP) were examined. Optimal conditions for autolysis were the same as those required for expression of activity of CANP. The autolysis proceeded rapidly by a strictly limited proteolysis. During autolysis the molecular weight of CANP changed from 82 K (native CANP or mCANP) to 79 K and then 60 K. The 79 K and 60 K molecular species were both active at microM order of Ca2+ (muCANP), whereas the native CANP is active at mM order of Ca2+ (mCANP). Various proteases examined did not produce muCANP from mcanp under the conditions tested. Furthermore, muCANP did not yield muCANP from mCANP at lower Ca2+ concentrations where only muCANP was active. Therefore, muCANP is produced from mCANP only by the specific autolysis of mCANP. PMID- 6277880 TI - The cytosol of human erythrocytes contains a highly Ca2+-sensitive thiol protease (calpain I) and its specific inhibitor protein (calpastatin). AB - The cytosol of human erythrocytes was found to contain a Ca2+-dependent thiol protease (calpain) and its specific inhibitor (calpastatin) by DEAE-cellulose chromatography at pH 8.0, although no proteolytic activity toward casein was detected in the unfractionated hemolysate. The protease required only 40 microM Ca2+ for 50% activation, indicating that it belongs to the highly Ca2+-sensitive type of calpain, namely, calpain I. It was not inactivated by heating at 58 degrees C for 10 min at pH 7.2, the optimal pH for its action on casein. The inhibitor comprised major and minor components, calpastatin H (Mr = 280,000) and caplastatin L (Mr = 48,000). Both were heat-stable proteins which were readily inactivated by tryptic digestion. The inhibition of erythrocyte calpain by erythrocyte calpastatin H or L was not due to sequestering of Ca2+ from the reaction medium by the inhibitor protein. The calpain preparation preferentially digests bands III and IVa of human erythrocyte membrane proteins, with little or no cleavage of the bands corresponding to spectrin. PMID- 6277881 TI - Vanadate binding to the (Na + K)-ATPase. AB - A particulate (Na + K)-ATPase preparation from dog kidney bound [48V]-ortho vanadate rapidly at 37 degrees C through a divalent cation-dependent process. In the presence of 3 mM MgCl2 the Kd was 96 nM; substituting MnCl2 decreased the Kd to 12 nM but the maximal binding remained the same, 2.8 nmol per mg protein, consistent with 1 mol vanadate per functional enzyme complex. Adding KCl in the presence of MgCl2 increased binding, with a K0.5 for KCl near 0.5 mM; the increased binding was associated with a drop in Kd for vanadate to 11 nM but with no change in maximal binding. Adding NaCl in the presence of MgCl2 decreased binding markedly, with an I50 for NaCl of 7 mM. However, in the presence of MnCl2 neither KCl nor NaCl affected vanadate binding appreciably. Both the nonhydrolyzable, beta, gamma-imido analog of ATP and nitrophenyl phosphate, a substrate for the K-phosphatase reaction that this enzyme also catalyzes, decreased vanadate binding at concentrations consistent with their acting at the low-affinity substrate site of the enzyme, the presence of KCl increased the concentration of each required to decrease vanadate binding. Oligomycin decreased vanadate binding in the presence of MgCl2, whereas dimethyl sulfoxide and ouabain increased it. With inside-out membrane vesicles from red blood cells vanadate inhibited both the K-phosphatase and (Na + K)-ATPase reactions; however, with the K-phosphatase reaction extravesicular K+ (corresponding to intracellular K+) both stimulated catalysis and augmented vanadate inhibition, whereas with the (Na + K) ATPase reaction intravesicular K+ (corresponding to extracellular K+) both stimulated catalysis and augmented vanadate binding. PMID- 6277883 TI - Estimation of lipid regions in a cytochrome oxidase-lipid complex using spin labeling electron spin resonance: distribution effects on the spin label. AB - The distribution of lipid in the cytochrome oxidase-lipid complex from beef heart mitochondria has been studied by the spin labeling electron spin resonance technique. The spectra of a phospholipid spin label incorporated in the complex reveals an immobilized (on the ESR time scale) component in addition to the fluid component which is found in aqueous dispersions of the extracted lipids. The first component corresponds to the domain of lipid influenced by the protein, and the second component to the remaining lipid. A theory taking into account not only the sizes of the lipid regions in which the spin label molecule distributes itself, but also the different affinities of the label for the two domains, has been developed. Taking advantage of the variation in spectra obtained with increasing amounts of spin label, computer calculations have been performed to estimate the distribution of lipid in the different regions of the cytochrome oxidase-lipid complex. An extrapolation of the amount of immobilized spin-labeled phospholipid to zero concentration of label allows a calculation of the number of fatty acid residues interacting with the protein to be made. It has been found that the number of aliphatic chains influenced by the protein is higher than that calculated for a single boundary layer around the protein. The approach used in this paper can be used for studies of protein-lipid interactions in other systems. PMID- 6277882 TI - Studies on the molecular basis of H+ translocation by cytochrome c oxidase. AB - We report here studies which characterize further the interaction of N,N' dicyclohexylcarbodiimide with cytochrome c oxidase leading to inhibition of H+ translocation by the enzyme. Further evidence is presented to show that the inhibition results from a real interaction of DCCD with the enzyme and cannot be accounted for by uncoupling and, contrary to recent criticisms, this interaction occurs specifically with subunit III of the enzyme even at relatively high inhibitor-to-enzyme stoichiometries. Use of a spin-label analogue of DCCD has enabled us to demonstrate that the carbodiimide-binding site is highly apolar and may not lie on the pathway of electron transfer. PMID- 6277884 TI - Photosystem I reaction centers from Chlamydomonas and higher plant chloroplasts. AB - A photosystem I reaction center has been isolated from Chlamydomonas chloroplasts and compared with the photosystem I reaction center from higher plants. While the higher plant reaction center is active in cytochrome 552 photooxidation, the Chlamydomonas preparation was not active unless salts were included in the assay medium or the pH was lowered to 5. Subunit III-depleted photosystem I reaction center from higher plants is also inactive in cytochrome 552 photooxidation in the absence of salts. As with the Chlamydomonas reaction center, salts induced its activity. Subunit I of the photosystem I reaction center has tentatively been identified as the binding site of cytochrome 552. PMID- 6277886 TI - Analysis of reversible lipoprotein-cell interactions. AB - Quantitation and characterization of freely reversible ligand-receptor interactions in adherent cell lines grown in vitro have been technically difficult to perform. We report the growth of human fibroblasts on a microcarrier bead system which obviates the limitation of cell adherence in such determinations. This system has been applied to the reversible interaction of low density lipoproteins (LDL) with the fibroblast membrane receptors. Of the 125I LDL specifically bound to filtered cells, 52.2 +/- 9.8% was reversibly bound. This reversible LDL-cell binding occurred in less than 1 h at 37 degrees C, was specific for LDL, and occurred without significant 125I-LDL degradation. Using these techniques, it is now possible to obtain direct measurement and characterization of lipoprotein-receptor interactions in adherent cell lines of normal subjects and patients with dyslipoproteinemia. PMID- 6277885 TI - Orientation of complex III in the yeast mitochondrial membrane: labeling with [125I] diazobenzenesulfonate and functional studies with the decyl analogue of coenzyme Q as substrate. AB - Mitochondria (or mitoplasts) and submitochondrial particles from yeast were treated with [125I] diazobenzenesulfonate to label selectively proteins exposed on the outer or inner surface of the inner mitochondrial membrane. Polyacrylamide gel analysis of the immunoprecipitates formed with antibodies against Complex III or cytochrome b revealed that the two core proteins and cytochrome b were labeled in both mitochondria and submitochondrial particles, suggesting that these proteins span the membrane. Cytochrome c1 and the iron sulfur protein were labeled in mitochondria but not in submitochondrial particles, suggesting that these proteins are exposed on the cytosolic side of the inner membrane. The steady-state reduction of cytochromes b and c1 was determined with succinate and the decyl analogue of coenzyme Q as substrates. Addition of the coenzyme Q analogue to mitochondria caused reduction of 15-30% of ;the total dithionite reducible b and 100% of the cytochrome c1: Addition of the coenzyme Q analogue to submitochondrial particles led to the reduction of 70% of the total dithionite reducible cytochrome b but insignificant amounts of cytochrome c1. A model to explain the topography of Complex III in the inner membrane is proposed based on these results. PMID- 6277887 TI - Controlled proteolysis by subtilisin as a probe for cyanide-induced conformational changes in Pseudomonas cytochrome oxidase. AB - Oxidized Pseudomonas cytochrome oxidase (ferrocytochrome c-551:oxidoreductase; EC 1.9.3.2), digested with subtilisin in the absence and presence of KCN, produces discrete, high molecular weight fragments. The presence of KCN alters the rate of this fragmentation but does not change the nature of the fragments. When digested in the absence of KCN, the oxidase gives a major product (A) which is enzymatically active and has an apparent Mr = 58,000 on sodium dodecyl sulfate polyacrylamide gel electrophoresis. In the presence of KCN, the major product (B) has Mr = 48,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis but with gel permeation high performance liquid chromatography it has an apparent Mr = 92,000. This implies that product B is dimeric, as is the parent enzyme which has Mr = 110,000 by high performance liquid chromatography. Absorption spectra of product B, isolated by gel filtration, show that it contains only the heme d1 moiety. The digestion time course indicates that the rate at which several minor products are formed is also dependent on the absence or presence of KCN. These observations suggest that the binding of KCN to the heme centers induces a conformational change in the enzyme so that the heme c-containing portion of the protein, which is at one end of the intact enzyme, can be removed without disrupting the integrity of the heme d1-containing portion. PMID- 6277888 TI - Membrane channel-forming polypeptides. Aqueous phase aggregation and membrane modifying activity of synthetic fluorescent alamethicin fragments. AB - Synthetic fragments of the membrane channel-forming polypeptide, alamethicin, have been labeled with a fluorescent 5-dimethylaminonaphthalene-1-sulfonyl (dansyl) group at the NH2-terminal. Seventeen and 13-residue fluorescent peptide esters have been shown to translocate divalent cations in unilamellar liposomes and uncouple oxidative phosphorylation in rat liver mitochondria. The corresponding peptide acids also exhibit membrane-modifying activity, whereas the shorter fragments are inactive. Aggregation of fluorescent peptides in aqueous solutions leads to a marked blue shift and enhancement of the dansyl group emission spectrum. "Critical micelle" concentrations may be determined for the association of peptides. The longer peptides aggregate at lower concentrations than the short peptides, with the ease of aggregation following a trend similar to that for functional activity. The peptide acids aggregate only in media of high ionic strength. The peptide ester aggregates are stabilized by increasing salt concentration and dissociated by urea, suggestive of hydrophobic stabilization of the aggregates. The enthalpy of association for the 10- and 17 residue peptide esters is estimated to be between -1 and -3 kcal mol-1 of monomer. The aqueous phase aggregation of channel-forming peptides at low concentrations suggests that preformed aggregates may be inserted into the membrane to constitute functional channels. PMID- 6277889 TI - (Na+,K+)-cotransport in the Madin-Darby canine kidney cell line. Kinetic characterization of the interaction between Na+ and K+. AB - Confluent monolayer cultures of the differentiated kidney epithelial cell line, Madin-Darby canine kidney cells (MDCK), have been used to study ion transport mechanisms involved in transepithelial transport. We have investigated the previously reported K+-stimulation of 22Na+ uptake by confluent monolayers of Na+ depleted cells (Rindler, M. J., Taub, M., and Saier, M. H., Jr. (1979) J. Biol. Chem. 254, 11431-11439). This component of Na+ uptake was insensitive to ouabain and amiloride, but was strongly inhibited by furosemide or bumetanide. Ouabain insensitive 86Rb+ uptake was also inhibitable by furosemide or bumetanide and stimulated by extracellular Na+. The synergistic effect of Na+ and 86Rb+ uptake and K+ on 22Na+ uptake was reflected by an increase in the apparent Vmax and a decrease in the apparent Km as the concentration of the other cation was increased. The extrapolated Km for either 86Rb+ or 22Na+ uptake in the absence of the other cation was 30 mM while the Km in the presence of a saturating concentration of the other cation was 9 mM. The absolute Vmax values for 22Na+ and 86Rb+ uptake suggest a cotransport system with a stoichiometry of 2Na+:3K+. However, because of the experimental design, the actual ratio may be closer to 1:1. Competition with, and stimulation by, a variety of unlabeled cations indicated that Na+ could be partially replaced by Li+, while K+ could be fully replaced by Rb+ and partially replaced by NH4+ and CS+. Uptake by this system was dependent upon cellular ATP. Reduction of intracellular ATP to 3% of normal abolished both K+-stimulated 22Na+ uptake and Na+-stimulated 86Rb+ uptake. PMID- 6277890 TI - Furosemide-sensitive salt transport in the Madin-Darby canine kidney cell line. Evidence for the cotransport of Na+, K+, and Cl-. AB - Confluent monolayer cultures of the Madin-Darby canine kidney (MDCK) cell line have been shown to possess a furosemide and bumetanide-sensitive (Na+,K+) cotransport system. We have studied the effect of anion substitutions on (Na+,K+) cotransport. In Na+-depleted cells, bumetanide-sensitive uptake of 22Na+ or 86Rb+ exhibited an absolute requirement for extracellular Cl-. Chloride could be replaced in the buffers by Br-, but not by F-, I-, acetate, nitrate, thiocyanate, sulfate, or gluconate. The effect of Cl- was saturating, and Na+-stimulated 86RB+ uptake as well as K+-stimulated 22Na+ uptake was shown to be dependent on the square of the Cl- concentration. The concentration of Cl- which gave half-maximal stimulation of cation cotransport varied between 58 and 70 mM. There was a small degree of cooperativity between the binding affinities for Cl- and K+ at constant Na+ concentrations. Bumetanide-sensitive 36Cl- uptake could be demonstrated when extracellular Na+ and K+ were present simultaneously. Uptake through this system was unaffected by changes in the membrane potential or by the imposition of pH gradients. Together these data strongly suggest that the bumetanide-sensitive transport system in Madin-Darby canine kidney cells co-transports Na+, K+, and Cl in a ratio of 1:1:2. PMID- 6277891 TI - Spin state and unfolding equilibria of ferricytochrome c in acidic solutions. AB - Equilibrium, stopped flow, and temperature-jump spectrophotometry have been used to identify processes in the unfolding of ferricytochrome c in acidic aqueous solutions. A relaxation occurring in approximately 100 microseconds involves perturbation of a spin-equilibrium between two folded conformers of the protein with methionine-80 coordinated or dissociated from the heme iron. The protein unfolds more slowly, in milliseconds, with dissociation and protonation of histidine-18. These two transitions appear cooperative in equilibrium measurements at low (0.01 M) ionic strength, but are separated at higher (0.10 M) ionic strength. They are resolved under both conditions in the dynamic measurements. The spin-equilibrium description permits a unified explanation of a number of properties of ferricytochrome c in acidic aqueous solutions. PMID- 6277892 TI - Interchain disulfide bonding in the regulatory subunit of cAMP-dependent protein kinase I. AB - The two protomers of the purified regulatory subunit from porcine cAMP-dependent protein kinase I have been shown to be covalently cross-linked by interchain disulfide bonding. Limited proteolysis which cleaves the polypeptide chain into two fragments demonstrated that the disulfide bonding was associated exclusively with the fragment that corresponded to the NH2-terminal region of the polypeptide chain. This NH2-terminal fragment accounted for approximately 15 to 20% of the molecule. The disulfide bonding was further characterized by alkylating the cysteines in the native regulatory subunit. Following oxidation with performic acid, each regulatory subunit contained 7 cysteic acid residues; however, under denaturing conditions, but without prior reduction, only 5 cysteine residues could be alkylated with iodoacetic acid. Following limited proteolysis, all five of these cysteines were associated with the larger COOH-terminal, cAMP binding domain. In contrast, if the denatured subunit was first reduced prior to alkylation, all 7 cysteine residues were alkylated. The 2 cysteines that were only accessible to alkylation after prior reduction were both associated with the NH2-terminal end of the polypeptide chain ultimately with a 5,400 peptide. Alkylation of the isolated, denatured NH2-terminal domain with iodoacetic acid resulted in no covalent modification unless the fragment was first reduced with dithiothreitol. The NH2-terminal and COOH-terminal domains were shown to be linked by a region of the polypeptide chain that is rich in both proline and arginine. It is the arginine-rich site that is readily prone to proteolytic cleavage. PMID- 6277893 TI - Role of actin and tubulin in the regulation of poly(A) polymerase endoribonuclease IV complex from calf thymus. AB - The poly(A) polymerase from calf thymus (Mr = 62,000) is associated with the poly(A)-specific endoribonuclease IV (Mr = 45,000); this complex is dissociable at pH 8.2. The activities of the two enzymes in the complex are strongly inhibited by the G-actin and tubulin (both in the subunit-dissociated and in the polymeric form); F-actin was less inhibitory, while myosin and actomyosin were without any effect. Gel filtration experiments revealed a complex formation between poly(A) polymerase/endoribonuclease IV and G-actin or tubulin at an approximately 1:1 molar ratio. The homogeneous poly(A) polymerase was determined to be maximally inhibited by a 5-fold excess of G-actin and by less than a 2-fold excess of tubulin dimer, if calculated on the molecular basis. The inhibitory potency of G-actin on poly(A) polymerase was reduced after co-incubation with DNase I or phalloidin and that of tubulin by colchicine. It is suggested that actin and/or tubulin is involved also in regulation of poly(A) metabolism in intact cells. PMID- 6277894 TI - Structural characterization of neutral oligosaccharides of human midcycle cervical mucin. AB - It was previously shown that alkaline borohydride treatment of human midcycle cervical mucin releases a heterogeneous population of reduced neutral, sialylated, and sulfated oligosaccharides (Yurewicz, E. C., and Moghissi, K. S. (1981) J. Biol. Chem. 256, 11895-11905). Three major neutral oligosaccharides were isolated with approximate compositions of Fuc:Gal:GlcNAc:N acetylgalactosaminitol (GalNAcol) = 0:2:1:1 (A1), 1:2:1:1 (A2), and 2:2:1:1 (A3). They comprised roughly 21%, 13%, and 8% of human cervical mucin oligosaccharide chains, respectively. In the present report, each was analyzed by periodate oxidation, methylation, and sequential degradation with glycosidases. A1 was shown to contain more than one component, but structural analyses clearly demonstrated the presence of one predominant (75%) tetrasaccharide. The proposed structure, Gal beta 1-4GlcNAc beta 1-6(Gal beta 1-3)GalNAcol, has previously been found in human gastric, submaxillary, and ovarian cyst mucins in their carbohydrate-to-protein linkage regions. beta-Galactosidase from Aspergillus niger selectively cleaved the Gal beta 1-4GlcNAc linkage in the intact tetrasaccharide. Enzymatic hydrolysis of the Gal beta 1-3GalNAcol linkage required prior removal of the Gal beta 1-4GlcNAc beta 1-unit attached to 0-6 of GalNAcol. The data for A2 indicated a mixture of two oligosaccharides, Gal beta 1 4,3(Fuc alpha 1-3,4)GlcNAc beta 1-6(Gal beta 1-3)GalNacol and Fuc alpha 1-2Gal beta 1-4GlcNac beta 1-6(Gal beta 1-3)-GalNacol, in an approximate molar ratio of 3 to 4:1, respectively. Two structures are consistent with the data obtained for A3: Fuc alpha 1-2Gal beta 1-4,3(Fuc alpha 1-3,4)GlcNAc beta 1-6(Gal beta 1 3)GalNAcol and/or Gal beta 1-4,3(Fuc alpha 1-3,4)GlcNac beta 1-6(Fuc alpha 1-2Gal beta 1-3)GalNacol. The results indicate that A1 represents the "core" tetrasaccharide of the larger human cervical mucin oligosaccharides A2 and A3. PMID- 6277895 TI - Human somatotropin. Selective removal with trypsin of residues 135-145 from the hormone molecule with no loss of biological activities. AB - A 11-residue segment, sequence positions 135-145, has been cleaved selectively from the human somatotropin molecule by limited digestion of the hormone with trypsin at pH 6.0 in 90% (v/v) glycerol. Differences between the tertiary structures of the native and trypsin-modified hormones have been demonstrated by circular dichroism spectroscopy. Despite these conformational differences, the biological properties of intact hormone and its modified form appeared to be very similar, as tested by two receptor-binding assays, radioimmunoassay, and the rat tibia test. PMID- 6277896 TI - Binding of the substrates and the allosteric inhibitor adenosine 5'-diphosphate to phosphoribosylpyrophosphate synthetase from Salmonella typhimurium. AB - The binding of the substrates, ATP and ribose-5-P, and the most effective inhibitor, ADP, to phosphoribosylpyrophosphate synthetase from Salmonella typhimurium was characterized using equilibrium dialysis of these compounds labeled with 32P. In the absence of ribose-5-P, ATP, ADP, and the ATP analogue alpha,beta-methylene ATP each bind cooperatively with half-saturation at 50 to 90 microM and Hill coefficients of 1.5 to 2. We propose that all three compounds bind at the same set of sites, which are presumably the active sites. When ribose 5-P was added, methylene ATP and ADP binding at these sites became tighter (Kd approximately 3 to 6 microM at 10 mM ribose-5-P) and lost its cooperativity. In the presence of ribose-5-P, ADP, but not methylene ATP, bound to a second site with half-saturation at approximately 150 microM and a Hill coefficient greater than 3. This result confirms the existence of an allosteric ADP site, which was previously postulated from kinetic studies (Switzer, R. L., and Sogin, D. C. (1973) J. Biol. Chem. 248, 1063-1073). Binding of ribose-5-P could not be detected in the absence of nucleotides, but it was readily measured in their presence. The apparent Kd of ribose-5-P varied from greater than 1 mM to approximately 5 microM as the concentration of either ADP or methylene ATP was increased from 0 to 2 mM. Inhibition of the enzyme by action of ADP at both active and allosteric sites could be observed kinetically. PMID- 6277897 TI - The reaction of hydrogen peroxide with the dimethyl ester heme derivative of cytochrome c peroxidase. AB - A modified cytochrome c peroxidase was prepared by reconstituting apocytochrome c peroxidase with protoheme in which both heme propionic acid groups were converted to the methyl ester derivatives. The modified enzyme reacted with hydrogen peroxide with a rate constant of (1.3 +/- 0.2) x 10(7) M-1 s-1, which is 28% that of the native enzyme. The reaction between the modified enzyme and hydrogen peroxide was pH-dependent with an apparent pK of 5.1 +/- 0.1 compared to a value of 5.4 +/- 0.1 for the native enzyme. These observations support the conclusion that the apparent ionization near pH 5.4, which influences the hydrogen peroxide cytochrome c peroxidase reaction is not due to the ionization of the propionate side chains of the heme group in the native enzyme. A second apparent ionization, with pK of 6.1 +/- 0.1, influences the spectrum of the modified enzyme which changes from a high spin type at low pH to a low spin type at high pH. PMID- 6277898 TI - Resonance Raman and electron paramagnetic resonance studies on oxidized and ferricyanide-treated Clostridium pasteurianum ferredoxin. Vibrational assignments from 34S shifts and evidence for conversion of 4 to 3 iron-sulfur clusters via oxidative damage. Vibrational assignments from 34S shifts and evidence for conversion of 4 to 3 iron-sulfur clusters via oxidative damage. AB - Resonance Raman spectra are reported for oxidized ferredoxin from Clostridium pasteurianum and for protein reconstituted with 34S2-, using 4579 A laser excitation. The spectra are of much higher quality than that previously reported, and the 34S shifts provide assignments of the Fe-S modes. After treatment with ferricyanide, the resonance Raman spectrum closely resembles that of the [3Fe-3S] protein, ferredoxin II from D. gigas; the 34S shifts aid in assignments of the [3Fe-3S] modes. The epr signal associated with the [3Fe-3S] cluster (g = 2.01) corresponds maximally to 0.80 spin/molecule. Anaerobic addition of excess sulfide to the reduced, ferricyanide-treated protein regenerates a [4Fe-4S]1+ epr spectrum, equivalent in intensity to the [3Fe-3S] signal. The ubiquitous occurrence of a g = 2.01 signal in preparations of [4Fe-4S] proteins can be attributed to variable amounts of [3Fe-3S] cluster generated by adventitious oxidation. The ready conversion of [4Fe-4S] to [3Fe-3S] clusters in C. pasterianum raises the possibility that some [3Fe-3S] proteins may actually arise by oxidative damage of [4Fe-4S] proteins during aerobic isolation. PMID- 6277899 TI - A new immunoreactive probe for the isolation and analysis of plasma membrane polypeptides. Synthesis and properties of isethionyl-3-(N-2,4-dinitrophenyl) aminopropioimidate. PMID- 6277900 TI - Mercuric reductase. Purification and characterization of a transposon-encoded flavoprotein containing an oxidation-reduction-active disulfide. AB - The flavoprotein mercuric reductase catalyzes the two-electron reduction of mercuric ions to elemental mercury using NADPH as an electron donor. It has now been purified from Pseudomonas aeruginosa PAO9501 carrying the plasmid pVS1. In this plasmid system, where the mer operon is on the transposon Tn501, mercuric reductase comprises up to 6% of the soluble cellular protein upon induction with mercurials. The purification is a rapid (two-step), high yield (80%) procedure. Anaerobic titrations of mercuric reductase with dithionite revealed the formation of a charge transfer complex with an absorbance maximum around 540 nm. Striking spectroscopic similarities to lipoamide dehydrogenase and glutathione reductase were observed. These two enzymes, which catalyze the transfer of electrons between pyridine nucleotides and disulfides, are flavoproteins which contain an oxidation-reduction-active cysteine residue at the active site. The expectation that mercuric reductase contains a similar electron acceptor was confirmed when it was shown that mercuric reductase has the capacity to accept four electrons per FAD-containing subunit, and that two thiols become kinetically titrable by 5,5'-dithiobis-(2-nitrobenzoate) upon reduction with NADPH. These are characteristic features of the disulfide reductase class of flavoproteins. Further similarities with at least one of these enzymes, lipoamide dehydrogenase, include the E/EH2 midpoint potential (-269 mV), fluorescence properties, and extinction coefficients of E and EH2. Preliminary observations relevant to an understanding of the mechanism of mercuric reductase are discussed. PMID- 6277901 TI - 1,25-Dihydroxyvitamin D3 receptors and functions in cultured pig kidney cells (LLC PK1). Regulation of 24,25-dihydroxyvitamin D3 production. AB - Although 1,25-dihydroxyvitamin D3 (1,25-(OH)2D3) regulates the renal metabolism of 25-hydroxyvitamin D3 (25-OH-D3), the mechanism is not well understood. The established pig kidney cell line, LLC PK1, was used to study this feedback regulation. These cells possess a receptor for 1,25-(OH)2D3 with a sedimentation coefficient of 3.2 S. Scatchard analysis of 1,25-(OH)2D3 binding to cell cytosol yielded an apparent Kd of 0.12 nM and an Nmax of 26 fmol/mg of cytosol protein. LLC PK1 cells respond to 1,25-(OH)2D3 by changes in the metabolism of 25-OH-D3. When incubated with 25-OH-[3H]D3, homogenates of untreated cells did not produce detectable 1,25-(OH)2[3H]D3 or 24,25-(OH)2[3H]D3. However, after treatment of cell monolayers with 1,25-(OH)2D3 for 8 h, homogenates converted substantial 25 OH-[3H]D2 substrate to 24,25-(OH)2[3H]D3. The appearance of this 24-hydroxylase activity in response to 1,25-(OH)2D3 was time- and dose-dependent. Half-maximal levels of enzyme activity were achieved with 0.13 nM 1,25-(OH)2D3, a concentration almost identical to the Kd of the 1,25-(OH)2D3 receptor. The stimulation of 24-hydroxylase activity was shown to be an induction event; treatment of monolayers with 13 nM 1,25-(OH)2D3 for a 4-h pulse was sufficient to induce maximal activity assayed at h. The presence of the transcriptional inhibitor, actinomycin D, during the 4-h pulse abolished the induction of 24 hydroxylase activity. These results demonstrate for the first time the presence of both 1,25-(OH)2D3 receptors and stimulation of 24-hydroxylase activity within the same established mammalian kidney cell line. PMID- 6277902 TI - Physiological desensitization of carbohydrate permeases and adenylate cyclase to regulation by the phosphoenolpyruvate:sugar phosphotransferase system in Escherichia coli and Salmonella typhimurium. Involvement of adenosine cyclic 3',5'-phosphate and inducer. AB - Adenylate cyclase and a number of carbohydrate transport systems are subject to regulation by the phosphoenolpyruvate:sugar phosphotransferase system. These sensitive carbohydrate transport systems are desensitized to regulation by the phosphotransferase system, and adenylate cyclase is deactivated when cells are grown in medium containing cyclic AMP. These effects are specific for cyclic AMP and are potentiated by the genetic loss of cyclic AMP phosphodiesterase. Inclusion in the growth medium of an inducer of a sensitive transport system also promotes desensitization of that particular transport system. Inducer-promoted desensitization is specific for the particular target transport system, while cyclic AMP-promoted desensitization is general and affects several systems. Desensitization of the permeases to regulation, and inactivation of adenylate cyclase, are slow processes which are blocked by chloramphenicol and are therefore presumably dependent on protein synthesis. Several sugar substrates of the phosphotransferase system are capable of regulating the sensitive carbohydrate transport systems. The evidence suggests that desensitization to this regulation does not result from a direct effect on the functioning of Enzyme I, a small heat-stable protein of the phosphotransferase system, HPr, or an Enzyme II of the phosphotransferase system, but specifically uncouples the permease systems from regulation. PMID- 6277903 TI - Abnormal lipoprotein receptor-binding activity of the human E apoprotein due to cysteine-arginine interchange at a single site. PMID- 6277904 TI - "Covalent affinity" purification of ubiquitin-activating enzyme. AB - We have previously described an enzyme that activates ubiquitin, the heat-stable polypeptide of the ATP-dependent proteolytic system from reticulocytes (Ciechanover, A., Heller, H., Katz-Etzion, R., and Hershko, A. (1981) Proc. Natl. Acad. Sci. U. S. A. 78, 761-765). It carries out ubiquitin-dependent PPi-ATP and AMP-ATP exchange reactions and binds to the activated polypeptide by a thiolester linkage. We describe here a procedure for the purification of this enzyme by its binding to ubiquitin-Sepharose. Binding of the enzyme to the affinity column requires ATP and Mg2+, and bound enzyme cannot be displaced by high salt but can be eluted by raising the pH, by increased concentrations of a thiol compound, or by the joint supplementation of AMP and pyrophosphate. Another form of the enzyme that cannot carry out AMP-ATP exchange (but catalyzes ubiquitin-dependent PPi-ATP exchange) does not bind to the affinity column. These data indicate that a covalent, possibly thiolester intermediate, is formed between the activating enzyme and Sepharose-bound ubiquitin. It is suggested designating this procedure of enzyme isolation "covalent affinity" chromatography. The purified enzyme has an apparent Mr = 210,000 and appears to be composed of two subunits of Mr = 105, 000. ATP-dependent binding of ubiquitin to the purified enzyme and to its subunit is demonstrated. PMID- 6277905 TI - Ubiquitin-activating enzyme. Mechanism and role in protein-ubiquitin conjugation. AB - Energy-dependent proteolysis in rabbit reticulocyte lysates has an absolute requirement for both ATP and ubiquitin that is believed due to the ATP-dependent covalent conjugation of ubiquitin (Ub) to substrate proteins as a signal event. Recently the enzyme responsible for activation of ubiquitin has been purified to near homogeneity by covalent affinity chromatography. In the present work we demonstrate that the enzyme catalyzes the net reaction (formula: see text); in which both an enzyme-bound COOH-terminal ubiquitin thiolester and a COOH-terminal ubiquitin adenylate are formed. The complex is sufficiently stable to be isolated by exclusion chromatography and contains ubiquitin/AMP in a ratio of 2. The presence of two forms of covalently linked ubiquitin provides a rationale for the observation of both ubiquitin-dependent ATP:PPi and ATP:AMP exchange. Selective derivatization of the thiol site with iodoacetamide results in a modified enzyme capable of catalyzing only ATP:PPi exchange and in forming only ubiquitin adenylate. Derivatization of the thiol site prevents conjugate formation in a reconstituted system, demonstrating that the ubiquitin thiolester is the proximal donor for subsequent conjugate formation. A three-step mechanism is proposed for ubiquitin activation by the enzyme prior to conjugation. PMID- 6277906 TI - Purification and characterization of normal and mutant forms of T4 endonuclease V. AB - Endonuclease V of bacteriophage T4 has been purified to physical homogeneity from T4D-infected Escherichia coli 1100. The enzyme, whose molecular weight is 16,000, possessed two distinct catalytic activities, a pyrimidine dimer-DNA glycosylase and an apurinic/apyrimidinic endonuclease. They acted on UV-irradiated poly(dA) . poly(dT) in a sequential manner; the glycosylase cleaved the N-glycosyl bond between the 5'-pyrimidine of a dimer and the corresponding sugar and then the endonuclease hydrolyzed a phosphodiester bond on the 3'-side of the apyrimidinic site. The 5'-termini thus generated were phosphorylated by T4 polynucleotide kinase only after they had been subjected to direct photoreversal and then treated with alkaline phosphatase. By using two phage mutants, uvs-5 and uvs-13, it was shown that occurrence of an amber mutation in the denV gene caused a simultaneous loss of the two activities. Suppression of the mutation of uvs-5 rendered both activities partially active. When the mutation of uvs-13 was suppressed, a mutant form of enzyme that possessed only a glycosylase activity was produced. This suggests that there are two distinct domains in a single enzyme, each of which corresponds to one of the activities. PMID- 6277907 TI - Ribose-1-P is the essential precursor for nucleic acid synthesis in animal cells growing on uridine in the absence of sugar. PMID- 6277908 TI - Sequence modeling using semisynthetic ribonuclease S. AB - We have designed and synthesized a model pentadecapeptide predicted to have the essential sequence information needed to form a stable and enzymatically active noncovalent complex with bovine pancreatic ribonuclease S-protein. The model peptide sequence, based on the conformational approach of simplifying the native sequence in a manner consistent with retention of essential noncovalent contacts and of secondary structure features, contained alanine at all positions except for Glu 2, Lys 7, Phe 8, Arg 10, His 12, and Met 13. The peptide was synthesized by the Merrifield solid phase method. The circular dichroism spectra of the purified model peptide in water and trifluoroethanol indicated a tendency to form an alpha-helical structure similar to that found for native S-peptide. The model peptide formed a stable complex with ribonuclease S-protein. With 12-fold excess of the peptide, the complex exhibited 36% of the specific activity of fully native ribonuclease S against the substrate cyclic cytidine 2':3'-monophosphate at pH 7.15. The dissociation constant of the model peptide for S-protein was found to be 1.1 x 10(-6) M, compared with 0.1 x 10(-6) M for native S-peptide. Crystals grown of the model peptide-S-protein complex were found to be isomorphous with those of native complex. The activity, stability, and structural integrity of the model complex verify the deductions made about essential sequence information in the NH2-terminal region of ribonuclease. Further, the results emphasize the general usefulness of the conformational approach in designing simplified sequences for other peptides and proteins. PMID- 6277909 TI - Purification of the low density lipoprotein receptor, an acidic glycoprotein of 164,000 molecular weight. AB - This paper describes a rapid two-step procedure for the purification of the low density lipoprotein receptor from bovine adrenal cortex membranes. After solubilization with nonionic detergents, the receptor adheres tightly to a DEAE cellulose column at pH 6. Following elution from DEAE-cellulose, detergent is removed, leaving the receptor in a soluble form. The receptor is then subjected to affinity chromatography on low density lipoprotein coupled to Sepharose 4B. The receptor is eluted with suramin, a newly-found inhibitor of low density lipoprotein-receptor interactions. This procedure yields a single protein with a molecular weight of 164,000. The same protein is also isolated when the crude DEAE-cellulose fraction is applied to an immunoaffinity column containing a monoclonal antibody directed against the receptor. The 164,000-dalton receptor protein has an acidic isoelectric point of 4.6, which rises to 4.8 after extensive treatment with neuraminidase. The purified receptor retains all of the binding properties of the receptor of intact cells and crude membranes. PMID- 6277910 TI - Catenation of DNA rings by topoisomerases. Mechanism of control by spermidine. PMID- 6277911 TI - Isolation and characterization of a human locus homologous to the transforming gene (v-fes) of feline sarcoma virus. AB - A single locus (designated c-fes) in the human genome which exhibits homology to the transformation-specific onc gene (v-fes) of Snyder-Theilen feline sarcoma virus was identified by the Southern blot technique. Recombinant clones containing 16- to 18-kilobase inserts of human DNA including the c-fes locus were constructed. Restriction endonuclease mapping of these clones verified their identity with native human c-fes and demonstrated the presence of at least two sequences in human c-fes interrupting v-fes-homologous regions. The v-fes homologous locus in the human genome spans about 4 kilobases. The 5'-3' orientation of the c-fes clones with respect to feline sarcoma virus proviral DNA was determined. The region of the human genome that is homologous to v-fes is proximal to the highly reiterated human Alu sequence but not to the highly reiterated human alphoid sequence. PMID- 6277912 TI - Human neutrophils transform prostaglandins by a myeloperoxidase-dependent mechanism. AB - Intact human neutrophils, incubated with the soluble stimulant phorbol myristate acetate, discharge lysosomal components, generate oxygen metabolites, and transform exogenous 6-keto-prostaglandin F1 alpha, prostaglandin E2, and prostaglandin F2 alpha as assessed by thin layer radiochromatography. Neutrophils alone were incapable of transforming the prostaglandins. The addition of catalase or the myeloperoxidase inhibitor, azide, protected all three prostaglandins from the phorbol-stimulated neutrophils. Neither superoxide dismutase, heat inactivated catalase, nor albumin had any inhibitory effect in this system. A model system consisting of glucose-glucose oxidase, as a source of H2O2, purified myeloperoxidase, and chloride was also able to transform the prostaglandins in an identical fashion. Neither glucose-glucose oxidase alone nor glucose-glucose oxidase and myeloperoxidase under chloride-free conditions were able to mediate this transformation. Thus, it appears that intact human neutrophils can transform prostaglandins by a mechanism dependent on H2O2, the lysosomal enzyme myeloperoxidase, and chloride. Given the importance of prostaglandins in regulating immune function, neutrophil-dependent prostaglandin transformation could play a novel role in modulating the inflammatory response. PMID- 6277913 TI - Preparation of a pure monoiodo derivative of the bee venom neurotoxin apamin and its binding properties to rat brain synaptosomes. AB - The preparation and purification of an active monoiodo derivative of apamin is described. Radiolabeled monoiodoapamin (2000 Ci/mmol) binds specifically to rat brain synaptosomes at 0 degrees C and pH 7.5 with a second order rate constant of association (ka = 2.6 x 10(7) M-1 s-1) and a first order rate constant of dissociation (kd = 3.8 x 10(-4) s-1). The maximal binding capacity is 12.5 fmol/mg of protein and the dissociation constant is 15-25 pM for the monoiodo derivative and 10 pM for the native toxin. The apamin receptor is destroyed by proteases suggesting that it is of a proteic nature. Neurotensin and its COOH terminal partial sequences are the only molecules unrelated to apamin that are able to displace monoiodoapamin from its receptor at low concentrations. Half displacement occurs at 170 nM neurotensin. This property is due to the presence in the COOH-terminal sequence of neurotensin of two contiguous arginine residues, a structure analogous to that of the apamin active site. The binding of monoiodoapamin to its receptor is sensitive to cations. Increasing K+ or Rb+ concentrations from 10 microM to 5 mM selectively enhances the binding by a factor of 1.8. Increasing the concentration of any cation from 1 to 100 mM completely inhibits iodoapamin binding. Both effects are due to a cation-induced modulation of the affinity of monoidoapamin for its receptor without any change of the maximal toxin binding capacity of synaptosomes. Guanidinium and molecules containing a guanidinium group are better inhibitors of iodoapamin binding than other inorganic cations or positively charged organic molecules. PMID- 6277914 TI - Conditions which cause the right-handed to left-handed DNA conformational transitions. Evidence for several types of left-handed DNA structures in solution. PMID- 6277915 TI - Uncoupling of alpha-adrenoceptor-mediated inhibition of human platelet adenylate cyclase by N-ethylmaleimide. AB - Human platelet adenylate cyclase is stimulated by prostaglandin E1 (PGE1) and is inhibited by epinephrine via alpha-adrenoceptors. Both agonists, epinephrine more than PGE1, increase the activity of a low Km GTPase in platelet membranes. Pretreatment of intact platelets or platelet membranes with the sulfhydryl reagent, N-ethylmaleimide (NEM), abolished the inhibition of the adenylate cyclase and the concomitant stimulation of the GTPase by epinephrine. In contrast, stimulation of the adenylate cyclase by PGE1 was not affected or even increased by NEM pretreatment; only at high NEM concentrations were both basal and PGE1-stimulated activities decreased. Similarly, the PGE1-induced activation of the low Km GTPase was not or was only partially reduced by NEM. Adenylate cyclase activation by stable GTP analogs, NaF, and cholera toxin was also not decreased by NEM pretreatment. Exposure of intact platelets to NEM did not reduce alpha-adrenoceptor number and affinities for agonists and antagonists, as determined by [3H]yohimbine binding in platelet particles. The data indicate that NEM uncouples alpha-adrenoceptor-mediated inhibition of platelet adenylate cyclase, leaving the receptor recognition site and the adenylate cyclase itself relatively intact. Although the effect of NEM may be based on a reaction with the alpha-adrenoceptor site interacting with a coupling component, the selective loss of the adenylate cyclase inhibition together with an even increased stimulation of the enzyme by PGE1 suggests that there are two at least partially distinct regulatory sites involved in opposing hormonal regulations of adenylate cyclase activity, with that involved in hormonal inhibition being highly susceptible to inactivation by NEM. PMID- 6277916 TI - On the recognition and cleavage mechanism of Escherichia coli endodeoxyribonuclease V, a possible DNA repair enzyme. AB - Escherichia coli endodeoxyribonuclease V acts at many sites of damage in duplex DNA, including apurinic/apyrimidinic sites, lesions induced by ultraviolet light which are not pyrimidine dimers, adducts of 7-bromomethylbenz[a]anthracene, and, as demonstrated earlier (Gates, F. T., and Linn, S. (1977a) J. Biol. Chem. 252. 1647-1653), it degrades uracil-containing duplex DNA most efficiently. The cleavage rate increases with increasing substitution of uracil for thymine in T5 DNA, with a replacement of one-eight of thymine generating the apparent maximum cleavage rate. However, the apparent reaction limit with DNA containing 3.8% of thymine replaced by uracil corresponds to cleavage at only 6% of the dUMP residues. Evidently, the enzyme recognizes some peculiarities of abnormal DNA structure, but not simply distortions, since some lesions, including pyrimidine dimers, are not substrates. Endonuclease V generates double strand breaks in a constant ratio to single strand nicks, regardless of the substrate. It degrades DNA processively, completing the digestion of one substrate molecule before proceeding to the next. The enzyme also appears to act cooperatively. Cleavage at methylbenz[a]anthracene adducts is usually or always 5' to the lesion. Endonuclease V seems well suited to act as a DNA repair enzyme, surveying the genome for structural distortions generated by lesions for which specific repair systems might not exist. PMID- 6277917 TI - Asialoglycoprotein receptors in hepatic regeneration. AB - The hepatic binding protein which is responsible for receptor-mediated endocytosis of desialylated glycoproteins undergoes cell-cycle dependent modulation of its expression. Endocytosis of desialylated orosomucoid by hepatocytes isolated from regenerating liver following two-thirds hepatectomy is reduced by 80% and returns to normal when regeneration is substantially complete. This decrease in endocytotic activity is due to selective loss of cell-surface associated receptor during liver regeneration. PMID- 6277918 TI - Neutrophil-mediated methemoglobin formation in the erythrocyte. The role of superoxide and hydrogen peroxide. AB - Human neutrophils incubated with phorbol myristate acetate oxidized hemoglobin within the intact erythrocyte by a mechanism dependent on cell-cell contact but independent of phagocytosis. Spectrophotometric examination of the erythrocyte lysates revealed that the major component formed was methemoglobin along with small amounts of a species with spectral characteristics similar to choleglobin. Methemoglobin formation was directly related to the neutrophil concentration and the time of incubation. The addition of superoxide dismutase or catalase modestly inhibited the formation of methemoglobin, while a combination of the enzymes provided the most dramatic protection. Methemoglobin of hydroxyl radical or hypochlorous acid scavengers. Apparently, either O2.- or H2O2 alone was capable of mediating methemoglobin formation in the intact erythrocyte. Maintenance of the intraerythrocytic hemoglobin in its oxygenated state or its derivatization to carbon monoxyhemoglobin markedly inhibited methemoglobin formation. Blockade of the anion channels in the intact erythrocyte with sulfonated stilbenes inhibited O2.- but not H2O2 from oxidizing intracellular hemoglobin. It appears that neutrophil-derived O2.- and H2O2 can cross the erythrocyte membrane through the anion channel or diffuse directly into the intracellular space and react with oxyhemoglobin or deoxyhemoglobin to form a mixture of hemoglobin oxidation products within the intact cell. PMID- 6277919 TI - In vitro transcription of vesicular stomatitis virus. Incorporation of deoxyguanosine and deoxycytidine, and formation of deoxyguanosine caps. AB - Detergent-disrupted vesicular stomatitis virus carried out in vitro transcription at a reduced rate in the presence of deoxyguanosine triphosphate. The transcripts annealed completely to excess vesicular stomatitis virus genomic RNA and consistent of the short leader RNA and even polyadenylated messenger RNA. Incubation with RNase T1 showed that the transcripts were resistant to cleavage. Nearest-neighbor analysis demonstrated that approximately two-thirds of the guanosine residues had been replaced by deoxyguanosine. The hybrid "deoxyguanosine-RNAs" carried cap structures which contained deoxyguanosine instead of guanosine. Competition experiments using both guanosine- and deoxyguanosine triphosphates indicated that GpppA and dGpppA cap structures were synthesized in approximately equal amounts at a ratio of 20 microM guanosine triphosphate to 50 microM deoxyguanosine triphosphate. Deoxyguanosine triphosphate was accepted by the polymerases of various strains and serotypes of vesicular stomatitis virus, demonstrating that its incorporation was a common characteristic. Deoxycytidine triphosphate could also substitute for cytidine triphosphate but to a lesser degree. Deoxyadenine, deoxyuridine-, and thymidine triphosphates were not or were very poorly accepted even at concentrations of 2 MM. PMID- 6277920 TI - beta-Endorphin and deletion peptides. A correlation of opiate receptor affinity with helix potential. PMID- 6277921 TI - Diadenosine tetraphosphate hydrolase from mouse liver. Purification to homogeneity and partial characterization. AB - An enzyme hydrolyzing diadenosine 5',5"'P1, P4-tetraphosphate (Ap4A) to AMP and ATP has been purified to apparent homogeneity from mouse liver cell extracts. The isolation procedure comprised ammonium sulfate precipitation, chromatography on Sephadex G-75. DEAE-cellulose, blue Sepharose and AMP-Sepharose. The enzyme is a single polypeptide chain with a native Mr = 64,000 with a Km of 1.66 microM and Vmax of 1.25 mumol/min. AMP, ADP, Ap4, GTP, Gp4, Ap3A, Ap5A, Gp3G, and Gp5G are noncompetitive inhibitors of the Ap4A hydrolase activity, whereas Gp4G inhibits Ap4A hydrolysis competitively with a Ki of 6 microM. Theophylline, caffeine, and isobutylmethylxanthine do not or only slightly inhibit Ap4A hydrolysis. Mitogenic factors have no effect on the enzymatic activity of Ap4A hydrolase, excluding that a direct influence of internalized mitogens on Ap4A degradation could be responsible for mitogen-dependent fluctuation of intracellular Ap4A pool sizes. PMID- 6277922 TI - The primary structure of the Saccharomyces cerevisiae gene for alcohol dehydrogenase. AB - The DNA sequence of the gene for the fermentative yeast alcohol dehydrogenase has been determined. The structural gene contains no introns. The amino acid sequence of the protein as determined from the nucleotide sequence disagrees with the published alcohol dehydrogenase isozyme I (ADH-I) sequence for 5 of the 347 amino acid residues. At least one, and perhaps as many as four, of these differences is probably due to ADH-I protein heterogeneity in different yeast strains and not to sequencing errors. S1 nuclease was used to map the 5' and 3' ends of the ADH-I mRNA. There are two discrete, mature 5' ends of the mRNA, mapping 27 and 37 nucleotides upstream of the translation initiating ATG. These two equally prevalent termini are 101 and 91 nucleotides, respectively, downstream from a TATAAA sequence. Analysis of the 3' end of ADH-I mRNA disclosed two minor ends upstream of the major poly(A) addition site. These three ends map 24, 67, and 83 nucleotides, respectively, downstream from the translation-terminating TAA triplet. The sequence AA-TAAG is found 28 to 34 nucleotides upstream of each ADH I mRNA poly(A) addition site. Sequence comparisons of these three 3' ends with those for four other yeast mRNAs yielded a 13-nucleotide consensus sequence to which TAAATAAGA is central. All of the known yeast poly(A) addition sites map at or near the A residue of a CTA site 25 to 40 nucleotides downstream from this consensus octamer. PMID- 6277923 TI - Resolution of high and low affinity epidermal growth factor receptors. Inhibition of high affinity component by low temperature, cycloheximide, and phorbol esters. PMID- 6277924 TI - Electron paramagnetic resonance studies of MN(II) complexes with myosin subfragment 1 and oxygen 17-labeled ligands. AB - Ligands in the first coordination sphere of Mn(II) in the complex of MnADP with myosin subfragment 1 from rabbit skeletal muscle have been investigated. EPR spectroscopy was used to detect superhyperfine coupling between unpaired electrons of the metal ion and the nuclei of oxygen atoms specifically labeled with oxygen 17. The results show that ADP is a beta-monodentate ligand for Mn(II) and that there are probably two water oxygens directly bound to Mn(II). The inhibitory complex of vanadate with subfragment 1 . MnADP was also investigated. Vanadate-dependent changes in the EPR spectra for enzyme-bound Mn(II) indicate that the coordination sphere of MN(II) changes upon binding of vanadate. ADP remains a beta-monodenate ligand in the complex and experiments with 17O-labeled water indicate that two oxygen atoms originally in water are ligands in the complex. However, the oxygens of vanadate equilibrate with those of water during sample preparation so that one of these ligands may be a vanadate oxygen. Three additional ligands, probably from the protein, are required to complete the sextet of ligands to Mn(II) in both complexes studied. PMID- 6277925 TI - The subcellular localization of enzymes of dolichol metabolism in rat liver. AB - Dolichyl phosphate is an intermediate in the glycosylation of N-glycosamidic linked glycoproteins in mammalian systems, and its availability may be a limiting factor in glycoprotein biosynthesis. The basic kinetics and subcellular distribution of enzymes which may influence the concentration of dolichyl phosphate in rat liver have therefore been investigated. These include dolichyl phosphate phosphatase, dolichol phosphokinase, dolichyl fatty acyl ester synthetase, GDP-mannose dolichyl phosphate mannosyl transferase, and UDP-glucose dolichyl phosphate glucosyl transferase. The specific activity of the enzymes was highest in the microsomes, except for dolichyl phosphate phosphatase and dolichyl fatty acyl ester synthetase, which were most concentrated in the plasma membrane and the cytosol fraction, respectively. The nuclei contained all of the enzyme activities while the mitochondria and cytoplasm were generally less active. The presence of both dolichol phosphokinase and dolichyl phosphate phosphatase in microsomes and nuclei, which contain the highest glycosyl transferase activities, may provide a means for direct enzymatic control of levels of dolichyl phosphate. PMID- 6277926 TI - Evidence for translated intervening sequences in the mitochondrial genome of Saccharomyces cerevisiae. AB - In yeast, the mitochondrial genes for subunit I of cytochrome oxidase (oxi3) and for apocytochrome b (cob) are known to be split. In some strains, the latter contains five intervening sequences, three of which coincide with clusters of mutational sites referred to in their order of transcription as the loci box3, 10, and 7, respectively. Mutations at the first of these result in the accumulation of novel, large polypeptides (apparent Mr congruent to 43,000) believed to originate from a fusion of sequences found in the NH2-terminal segment of apocytochrome b to others encoded in the intervening sequence itself. We now provide evidence for close similarities of at least a part of translated intron sequences between (a) mutants in box7 in "long" form and "short" form strains (which lack the first three introns including the one for the box3 locus); (b) mutants in a subset of box7 mutants and those in box3, and (c) between intron sequences in box7 and a sequence presumably encoded in oxi3. These structural homologies have been analyzed and shown to be referable to sequence homologies in two proteins, one derived from the second intron (box3) in cob and the other from oxi3. The accumulation in certain cob mutants of proteins and of a transcript containing a sequence specified by oxi3 provides additional strong evidence for the previously suggested regulation of oxi3 by the penultimate, box7 containing intron of cob. PMID- 6277927 TI - Phosphorylation of glycogen synthase by cyclic AMP-independent casein kinase-2 from rabbit skeletal muscle. AB - Cyclic AMP-independent casein kinase-2 from rabbit skeletal muscle has been extensively purified by procedures including phosphocellulose, Bio-Gel A-1.5m, casein-Sepharose, and DEAE-cellulose column chromatographies. The casein kinase and glycogen synthase kinase activities are co-purified throughout the purification. The casein kinase-2 has Mr approximately equal to 135,000 as determined by glycerol density gradient centrifugation and by gel filtration. Analysis of the purified kinase by gel electrophoresis in the presence of sodium dodecyl sulfate reveals the presence of two major protein bands having Mr = 42,000 and 27,000 in a ratio of 0.85:1. The kinase phosphorylates glycogen synthase, casein, and phosvitin either in the presence of ATP or GTP; however, the Km values for GTP are slightly higher than those of ATP. The activity of this kinase is inhibited by heparin but is not affected by the addition of cyclic AMP, heat-stable inhibitor of cyclic AMP-dependent protein kinase, Ca2+, ethylene glycol bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid, or the combination of Ca2+ and calmodulin. The phosphorylation of the synthase by the kinase results in the incorporation of approximately 0.8 mol of PO4/subunit and a reduction in the synthase activity ratio from 0.82 to 0.6. The extent of phosphorylation of the synthase catalyzed by casein kinase-2 is similar to that by phosphorylase kinase; however, the sites phosphorylated by these two kinases are different. The relationships of casein kinase-2 to the cyclic AMP-independent synthase kinases that have been described by other laboratories and to the casein kinases isolated from other tissues are discussed. PMID- 6277928 TI - The cooperative role of the transformation-sensitive glycoproteins, GP140 and fibronectin, in cell attachment and spreading. AB - The detergent-insoluble matrix of cultured human fibroblasts contains cytoskeletal and nuclear components, as well as two major, noncollagenous glycoproteins, fibronectin and GP140. These glycoproteins are stabilized by extensive intermolecular disulfide bonding. GP140, in contrast to fibronectin, is resistant to digestion with trypsin and is not cross-reactive with antisera prepared against fibronectin (Carter, W. G., and Hakomori, S. (1981) J. Biol. Chem. 256, 6953-6960). GP140 was partially purified, under nonreducing conditions, by differential extraction of trypsinized cells with sodium trichloroacetate. Alternatively, a higher yield of GP140 could be obtained under reducing conditions by extraction with urea-dithiothreitol followed by molecular sieve chromatography in the presence of sodium dodecyl sulfate and dithiothreitol. The purified GP140 contained mannose, galactose, and N acetylglucosamine residues, totaling 2.7% of the molecule. In addition, mild periodate oxidation of GP140 followed by reduction with NaB3H4 under conditions designed to label sialic acid also labeled the peptide portion of the molecule. Amino acid analysis of GP140 detected periodate-sensitive hydroxylysine residues, as well as hydroxylproline, accounting for the periodate/NaB3H4-induced label in the peptide. These hydroxylated amino acids are major components of collagens and collagen-like proteins. The GP140 isolated under nonreducing conditions was found to induce stable cell attachment and cell spreading when coated on plastic surfaces. The cell attachment could not be inhibited with affinity purified anti fibronectin antibodies. However, trypsinization of cells under conditions that removed surface fibronectin reduced the ability of the cells to bind to the GP140 coated surface. Metabolic labeling of cells with radioactive glucosamine during 1 h cell attachment experiments incorporated label into both fibronectin and GP140, as well as four other carbohydrate-containing components as part of a stable detergent-insoluble matrix, indicating that the cells rapidly glycosylate both fibronectin and GP140 and incorporate them into the matrix. Long term labeling and chase experiments indicated that fibronectin and GP140 in the matrix are subject to very slow turnover. Neither fibronectin nor GP140 were detectable in the detergent-insoluble matrix of SV40-transformed human fibroblasts by either metabolic or cell surface labeling. These results are consistent with the conclusion that fibronectin and GP140 may function in a cooperative manner in cell adhesion and spreading. PMID- 6277929 TI - Euglena gracilis chloroplast transfer RNA transcription units. I. Physical map of the transfer RNA gene loci. AB - The locations of transfer RNA genes with respect to the restriction endonuclease cleavage map of Euglena gracilis Klebs, strain Z Pringsheim chloroplast DNA have been determined. Purified chloroplast tRNAs were treated with snake venom phosphodiesterase to remove the 3'-CCA terminus, and radioactively labeled by the action of Escherichia coli tRNA nucleotidyltransferase in the presence of [alpha 32P]CTP. Chloroplast DNA was treated individually and with combinations of the enzymes Bal I, Bam HI, Eco RI, Pst I, Pvu II, Sal I, and Xho I. The location of tRNA genes with respect to the cleavage sites for these enzymes was determined by hybridization of the 32P-labeled tRNAs to membrane filter blots of the chloroplast DNA restriction nuclease fragments following gel electrophoresis. The 145-kilobase pair genome was resolved into nine areas of strong tRNA hybridization, separated by areas of weak or no tRNA hybridization. The loci of tRNA genes are within the Eco RI fragments Eco A, B, G, H, I, J', P, Q, and V. PMID- 6277930 TI - Euglena gracilis chloroplast transfer RNA transcription units. II. Nucleotide sequence analysis of a tRNAVal-tRNAAsn-tRNAArg-tRNALeu gene cluster. AB - The tRNA-coding locus of the 8.2-kilobase pair (kbp) Eco RI fragments Eco G of Euglena gracilis Klebs, strain Z Pringsheim chloroplast DNA was chosen for detailed analysis. Two recombinant plasmids, pPG14, containing Eco G and the vector pMB9, and pEZC23, containing the chloroplast DNA fragment HindIII B cloned in pBR322 were employed for the study. The tRNA locus was mapped to an 0.8-kbp region of Eco G also present in HindIII B. The DNA sequence of a 1.6 kbp from HindIII B, containing the entire tRNA gene locus was determined. Four tRNA genes were identified from the DNA sequence. The gene organization is tRNAVal-16 bp spacer-tRNAAsn-3 bp spacer-tRNAArg-45 bp spacer-tRNALeu. The tRNALeu gene is of the opposite polarity as the other three genes. This is the first evidence of such a tRNA cluster for a chloroplast genome. Also evident from the DNA sequence, 132 bp from the 5'-end of the tRNALeu gene, is a putative gene or pseudogene for a chloroplast protein. PMID- 6277931 TI - Molecular characterization of the cAMP-dependent protein kinase bound to microtubule-associated protein 2. PMID- 6277932 TI - Thyrotropin-releasing hormone and cyclic AMP activate distinctive pathways of protein phosphorylation in GH pituitary cells. AB - The studies reported here were undertaken to clarify the cellular mechanism of the hypothalamic tripeptide, thyrotropin-releasing hormone (TRH), in clonal, hormone-responsive GH pituitary cells and to assess the possibility of a role for cyclic AMP as a mediator of TRH action. We investigated patterns of protein phosphorylation by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography of high speed supernatant and pellet fractions from untreated and treated GH cells. Brief treatment of cells with agents which elevate or mimic cellular cyclic AMP (8-bromo cyclic AMP, dibutyryl cyclic AMP, vasoactive intestinal polypeptide or cholera toxin) stimulated the phosphorylation of five supernatant peptides (41, 45, 47, 72, and 82 kilodaltons) and one pellet peptide (135 kilodaltons) and decreased the phosphorylation of one supernatant peptide (55 kilodaltons). In contrast, TRH promoted the phosphorylation of four different supernatant peptides (two 59, 65, and 80 kilodaltons). In addition, TRH also stimulated the phosphorylation of cyclic AMP-responsive 41-, 45-, and 82 kilodalton supernatant peptides and 135-kilodalton pellet protein and decreased the phosphorylation of 55-kilodalton supernatant peptide. Altered labeling of 47- and 72-kilodalton supernatant peptides, however, was not observed with TRH. Time course studies, as well as the overlapping biological action of TRH and vasoactive intestinal polypeptide, lead us to conclude that these peptide hormones utilize distinct, parallel pathways which converge at some late step. Furthermore, the results indicate that effects of TRH are mediated by a cyclic AMP-independent pathway. PMID- 6277933 TI - Host-dependent phosphorylation and kinase activity associated with vesicular stomatitis virus. AB - Among the protein kinases associated with vesicular stomatitis virus (VSV), one was identified by immunoprecipitation to be pp60src, the transformation-specific product coded for by avian sarcoma virus, or its endogenous cellular homolog. This activity phosphorylated only tyrosine. pp60src was enriched in the membranes, whereas the serine- and threonine-specific kinases were concentrated with viral cores. The content of pp60src in VSV can be manipulated by growing VSV in different host cells. Monolayer baby hamster kidney cells transformed by an avian sarcoma virus produced VSV progeny which contained 7-fold greater pp60src activity than progeny produced by control untransformed or revertant cells. In contrast, suspension cultures of baby hamster kidney cells which produced VSV with increased tyrosine-specific kinase activity did not affect the content of pp60src. When pp60src was specifically increased in cells, the endogenous phosphorylation of tyrosine residues in the VSV matrix M protein was also enhanced, to as much as 20-fold. The phosphorylation of serine or threonine in this protein or in the other VSV phosphoprotein NS was not affected. Cellular tyrosine-specific kinases other than pp60scr did not change the overall phosphorylation pattern of any VSV phosphoproteins. Experiments designed to test the effects of endogenous phosphorylation on the various functions of the M protein failed to detect any significant alterations. PMID- 6277934 TI - Preliminary diffraction data for crystals of ribonucleases A and B and their complexes with deoxy(pA)4 and deoxy(pA)6. AB - Crystals suitable for three-dimensional x-ray diffraction analysis have been reproducibly grown of both pancreatic ribonuclease A and B, and also of their complexes with deoxy-(pA)4 and deoxy-(pA)6. The crystals, all of which are grown from polyethylene glycol 4000 at low ionic strength, diffract to high resolution and would seem to provide another opportunity for the direct visualization of a protein-nucleic acid interaction. PMID- 6277935 TI - Streospecific substitution of oxygen-18 for sulfur in nucleoside phosphorothioates. AB - Reaction of nucleoside phosphorothioates with N-bromosuccinimide in dioxane and H218O leads to the exchange of sulfur for oxygen-18. Using the Sp-isomers of adenosine 5'-O-(1-thiodiphosphate) and adenosine 3',5'-cyclic phosphorothioate, it can be shown by 31P NMR spectroscopy that this reaction proceeds with inversion of configuration yielding the Rp-isomers of [alpha-18O]ADP and [18O]cAMP, respectively. Adenosine 5'-O-(2-thiotriphosphate) and adenosine 5'-O (3-thiotriphosphate) are likewise converted to [beta-18O]ATP and [gamma-18O]ATP although the stereochemistry of the former reaction has yet to be evaluated. With very slight modifications this reaction is applicable to all the common bases. PMID- 6277936 TI - Locking of hormone in the beta-adrenergic receptor by attack on a sulfhydryl in an associated component. AB - Isoproterenol incubated with turkey erythrocyte membranes causes exposure of a specific --SH in a component associated with the beta-adrenergic receptor, presumably in the guanyl nucleotide binding protein. Addition of a reagent which interacts with that specific --SH results in trapping of the hormone in the receptor. As a consequence, the number of hormone binding sites and the function of the receptor are both drastically reduced. Extended incubation at alkaline pH or addition of GDP or GTP at high concentration reactivate the beta-adrenergic receptor. Labeled antagonist binding as well as function of the receptor in activating an adenylate cyclase system are restored. The findings suggest that the normal interaction of the hormone-receptor complex with the guanyl nucleotide binding protein involves a conformational change which transiently locks the hormone in the receptor. GTP releases the tight interaction while addition of an -SH reagent traps the ternary complex of hormone-receptor-guanyl nucleotide binding protein in the locked conformation. Since the components of different hormone-activated adenylate cyclase systems were shown to be interchangeable, it seems likely that the hormone-receptor interaction with the guanyl nucleotide binding protein, as revealed in the present study, is not limited to beta adrenergic receptor systems. PMID- 6277937 TI - Cardiac phosphorylase kinase. Modulation of the activity by cAMP-dependent and cAMP-dependent phosphorylation of the alpha- subunit. PMID- 6277938 TI - Nucleotide sequence of Escherichia coli purF and deduced amino acid sequence of glutamine phosphoribosylpyrophosphate amidotransferase. AB - The Escherichia coli gene purF, coding for 5-phosphoribosylamine:glutamine pyrophosphate phosphoribosyltransferase (amidophosphoribosyltransferase) was subcloned from a ColE1-purF plasmid into pBR322. Amidophosphoribosyltransferase levels were elevated more than 5-fold in the ColE1-purF plasmid-bearing strain compared to the wild type control, and a further 10- to 13-fold elevation was observed in several pBR322 derivatives. The nucleotide sequence of a 2478-base pair PvuI-HinfI fragment encoding purF was determined. The purF45 structural gene codes for a 56,395 Mr protein chain having 504 amino acid residues. Methionine-1 is removed by processing in vivo leaving cysteine as the NH2-terminal residue. The deduced amino acid sequence was confirmed by comparisons with the NH2 terminal amino acid sequence determined by automated Edman degradation (Tso, J. Y., Hermodson, M. A., and Zalkin, H. (1982) J. Biol. Chem. 257, 3532-3536) and amino acid analyses of CNBr peptides including a 4-residue peptide from the CO2H terminus of the enzyme. Nucleotide sequences characteristic of bacterial promoter operator regions were identified in the 5' flanking region. The coding region appears to be preceded by a 277-297 nucleotide mRNA leader. A deletion removing the putative promoter-operator region results in defective purF expression. PMID- 6277939 TI - Spectroscopic investigations of ferric cytochrome P-450-CAM ligand complexes. Identification of the ligand trans to cysteinate in the native enzyme. AB - An extensive series of ligand complexes of ferric cytochrome P-450-CAM has been examined by UV-visible absorption, magnetic circular dichroism, and electron paramagnetic resonance spectroscopy in an attempt to identify the ligand trans to cysteinate in the six-coordinate resting state of the enzyme. Thus, the ligands used have been chosen to serve as models for coordination by potential endogenous amino acids and include alcohol, amide and carboxylate oxygen donors, amine, imidazole and indole nitrogen donors and disulfide, thioether, thiol, and thiolate sulfur donors. As this investigation has been by nature an empirical one, the conclusions are strengthened by the concurrent use of three different spectroscopic techniques. All of the complexes formed except those resulting from thiolate addition display spectroscopic properties that are broadly similar to those of low spin, six-coordinate P-450. Of the sulfur donor adducts, disulfide and thioether-bound P-450 have properties that are different enough in detail to distinguish them from native P-450. While the spectral features of the thiol bound species and of low spin ferric P-450 are alike, the former are pH dependent due to interconversion to bound thiolate, whereas the latter display essentially no spectral changes with pH. Of the oxygen donor complexes, all but carboxylate have spectra that very closely match those of the resting enzyme. Adducts formed with most nitrogenous ligands, including several imidazole derivatives, exhibit spectra that are sufficiently different from native P-450 to exclude them as candidates for the sixth ligand. Interestingly, the spectral properties of a complex formed with an imidazole derivative having a bulky electron-withdrawing substituent in the alpha position are comparable to those native P-450 except for the line shape of the EPR spectrum. Previously published theoretical work suggests that the spectral differences seen between this imidazole derivative and the other examined are electronic and not steric in origin. As no similar electronic mechanism exists for the protein to reduce the electron-donating ability or histidine, it is felt that coordination of histidine in the sixth position of P-450 can be ruled out. In conclusion, close examination of all spectral data reveals that amino acid analog adducts of P-450-CAM with amides and, in particular, alcohols, produce spectra that almost exactly duplicate those of native P-450 and suggests that the ligand trans to cysteinate in the six coordinate ferric enzyme has an oxygen donor atom. PMID- 6277940 TI - Effects of cholestyramine on low density lipoprotein binding sites on liver membranes from rabbits with endogenous hypercholesterolemia induced by a wheat starch-casein diet. AB - Rabbits fed a wheat starch-casein diet develop a marked hypercholesterolemia with a lipoprotein distribution similar to that of humans. Approximately 76% of the total cholesterol is carried in the low density lipoprotein (LDL) fraction (1.006 less than d less than 1.063 g/ml). Inclusion of 1% cholestyramine in the diet prevents the increase in plasma cholesterol. The cholestyramine effect is mediated through an increased fractional catabolic rate of 125I-LDL. In order to determine the potential role of hepatic LDL receptors in the removal of LDL from the plasma, binding of 125I-LDL and 125I-beta-VLDL (beta-migrating very low density lipoproteins) to hepatic membranes prepared from livers of rabbits fed the wheat starch-casein diet with or without cholestyramine supplementation was investigated. Membranes from livers of the cholestyramine-supplemented animals exhibit high levels of specific EDTA-sensitive binding of either of the 125I labeled lipoproteins. Very little EDTA-sensitive binding occurs on liver membranes from wheat starch-casein-fed rabbits that have not been treated with cholestyramine. These results indicate that the hypercholesterolemia in rabbits associated with the wheat starch-casein diet is wholly or partially the result of a decreased number of specific hepatic LDL receptors and thus a decreased catabolism of plasma cholesterol. The response of the liver to the inclusion in the diet of the bile acid sequestrant, cholestyramine, is to maintain or increase the number of specific LDL binding sites, thus promoting catabolism of plasma cholesterol. PMID- 6277941 TI - Ligand orientation of oxyproto- and oxymesocobalt porphyrin-substituted myoglobin by single crystal EPR spectroscopy. AB - Single crystals of oxyproto- and oxymesocobalt myoglobin have been examined by electron paramagnetic resonance spectroscopy at ambient and cryogenic temperatures in order to determine the principal values and eigenvectors of g tensors and the hyperfine coupling tensors. The Co--O--O bond angle was determined to be 125 degrees +/- 5 degrees for oxyprotocobalt myoglobin, and 153 degrees +/- 5 degrees for oxymesocobalt myoglobin at ambient temperature. This result suggests that differences in stereochemical interactions of the modified 2,4-side chains of porphyrin with protein contribute to the ligand orientations as well as the altered ligand-binding behavior in these hemoproteins. Upon freezing, two unequivalent orientations of the O--O axis (species I and II) were found in both oxycobalt myoglobin single crystals. Shifts of the resonance spectra of these species were observed below the freezing point of the crystals. The signal intensities of two paramagnetic species in oxyprotocobalt myoglobin were approximately equivalent (I congruent to II), whereas those in oxymesocobalt myoglobin were quite different (I greater than II) at 77 K. The present electron paramagnetic resonance studies demonstrate that changes in the bonding structure of Co--O2 are induced upon freezing the biological macromolecule, including the movement of the residues of the heme environment. PMID- 6277942 TI - Phosphorylase kinase specificity. A comparative study with cAMP-dependent protein kinase on synthetic peptides and peptide analogs of glycogen synthase and phosphorylase. AB - A synthetic pentadecapeptide, Pro-Leu-Ser-Arg-Thr-Leu-Ser-Val-Ser-Ser-Leu-Pro-Gly Leu-Glu, corresponding to the phosphorylatable site at the NH2 terminus of glycogen synthase, could be phosphorylated stoichiometrically at seryl residue 7 by both phosphorylase kinase and cAMP-dependent protein kinase. Phosphorylation of seryl residue 3 also occurred after prolonged incubation with cAMP-dependent protein kinase. Kinetic studies show that the pentadecapeptide is a better substrate for phosphorylase kinase. A peptide consisting of residues 1-11 was not as good a substrate and substitution of Arg-4 by Lys and Ser-9 by ARg in the unidecapeptide decreased and increased phosphorylase kinase reaction rates, respectively. Higher rates of phosphorylation were obtained with peptides of the phosphorylatable site of phosphorylase. A peptide with the sequence, Leu-Ser-Tyr Arg-Arg-Tyr-Ser-Leu was phosphorylated initially by phosphorylase kinase and cAMP dependent protein kinase at Ser-2 and Ser-7, respectively. Upon longer incubation, second site phosphorylation occurred with both kinases. A peptide of the same sequence with D-amino acids could not be phosphorylated but was a competitive inhibitor of both enzymes. The results suggest that optimal interaction of the two kinases depends on various factors including the orientation of arginyl groups with respect to the phosphorylatable serine. PMID- 6277943 TI - The horseradish peroxidase-catalyzed oxidation of 3,5,3',5'-tetramethylbenzidine. Free radical and charge-transfer complex intermediates. AB - Benzidine and related compounds are well known substrates for horseradish peroxidase/H2O2 oxidation. Typically, two different colored products are formed. In this paper, we study the oxidation of 3,5,3',5'-tetramethylbenzidine. The first colored product is a blue charge-transfer complex of the parent diamine and the diimine oxidation product. This species exists in rapid equilibrium with the radical cation. The radical was observed by ESR spectroscopy, and hyperfine splitting constants were determined. Addition of equimolar hydrogen peroxide yields the yellow diimine, which is stable at acid pH. At less than equimolar peroxide, all four species (diamine, radical cation, charge-transfer complex, and diimine) exist in equilibrium. A theoretical analysis of this redox system is presented, including a determination of the extinction coefficients and equilibrium constant for the nonradical species. PMID- 6277944 TI - Inhibition of epidermal growth factor binding to mouse cultured cells by fibroblast-derived growth factor. Evidence for an indirect mechanism. AB - Fibroblast-derived growth factor (FDGF), a basic, heat- and acid-stable polypeptide partially purified from the serum-free conditioned medium of BHK cells transformed by simian virus 40, is a potent mitogen for Swiss 3T3 cells and causes a marked reduction in 125I-labeled epidermal growth factor (125I-EGF) binding to these cells. The activity which inhibits EGF binding coelutes with the growth-stimulating activity after gel filtration, ion exchange chromatography, and sodium dodecyl sulfate polyacrylamide gel electrophoresis. Both cellular responses are elicited by the same range of FDGF concentration in several murine cell types. The inhibition of EGF binding is rapid and results from a decrease in the apparent affinity of cellular receptors for 125I-EGF. FDGF does not affect the rate of cell-mediated 125I-EGF degradation. Several lines of evidence suggest that FDGF does not bind directly to EGF receptor. First, the effect of FDGF is dependent on the temperature of the assay; furthermore, treatment of cells with EGF results in loss of EGF receptors while exposure to FDGF for up to 24 h does not induce "down-regulation" of EGF receptors. Further, in A431 cells which display a large number of specific EGF receptors, 125I-EGF binding is not sensitive to FDGF. Finally, the effect of FDGF on 125I-EGF binding is not observed with isolated plasma membranes. Taken together, these findings suggest that FDGF binds to sites which are separate from EGF receptors. The results show a novel mechanism whereby a growth-promoting factor produced by a tumor cell line can rapidly modulate the affinity of the cellular receptors for EGF in an indirect manner. PMID- 6277945 TI - The sodium/proton antiporter is part of the pH homeostasis mechanism in Escherichia coli. PMID- 6277946 TI - Function of the iron-sulfur protein of the cytochrome b-c1 segment in electron transfer reactions of the mitochondrial respiratory chain. AB - Resolution and reconstitution has been used to examine the involvement of the iron-sulfur protein of the cytochrome b-c1 segment in electron transfer reactions in this region of the mitochondrial respiratory chain. The iron-sulfur protein is required for electron transfer from succinate and from ubiquinol to cytochrome c1. It is not required for reduction of cytochrome b under these conditions, but it is required for oxidation of cytochrome b by cytochrome c plus cytochrome c oxidase. Removal of the iron-sulfur protein from the b-c1 complex prevents reduction of both cytochromes b and c1 by succinate or ubiquinol if antimycin is added to the depleted complex. As increasing amounts of iron-sulfur protein are reconstituted to the depleted complex, the amounts of cytochromes b and c1 reduced by succinate in the presence of antimycin increase and closely parallel the amounts of ubiquinol-cytochrome c reductase activity restored to the reconstituted complex, measured before addition of antimycin. The function of the iron-sulfur protein in these oxidation-reduction reactions is consistent with a cyclic pathway of electron transfer through the cytochrome b-c1 complex, in which the iron-sulfur protein functions as a ubiquinol-cytochrome c1/ubisemiquinone cytochrome b oxidoreductase. PMID- 6277947 TI - Photoaffinity labeling of the canine renal receptor for parathyroid hormone. AB - Studies were undertaken to identify and characterize components of the parathyroid hormone receptor. An analog of the bovine parathyroid hormone(1-34) sequence was derivatized at the 23-tryptophan position with the photoreactive reagent 2-nitro-5-azidophenylsulfenyl chloride. 2-Nitro-5-azidophenylsulfenyl bovine parathyroid hormone (NAPS-bPTH) analog retained full biological activity with respect to receptor binding and activation of adenylate cyclase in canine renal cortical plasma membranes. 125I-bPTH(1-34) analog was also derivatized without loss of receptor-binding activity. When 125I-NAPS-bPTH(1-34) analog was incubated with canine renal plasma membranes and then photolyzed, at least two 125I-labeled membrane components were identified by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The incorporation of 125I radioactivity into one of these components (Mr congruent to 60,000) was inhibited when incubation and photolysis were performed in the presence of excess, unlabeled bPTH(1-34). Furthermore, photolysis of membranes in the presence of NAPS-bPTH(1-34) analog led to activation of adenylate cyclase which persisted following washing to remove noncovalently bound peptide, suggesting a functional, covalent hormone receptor complex had been formed. We conclude that the M r congruent to 60,000 membrane component may be a part of the renal receptor for parathyroid hormone. PMID- 6277948 TI - Modulation by islet-activating protein of adenylate cyclase activity in C6 glioma cells. AB - The cAMP content of intact cells as well as adenylate cyclase of the membrane rich particulate fractions was studied with C6 glioma cells that had been exposed to the culture medium supplemented with islet-activating protein (IAP), one of the pertussis toxins. Both the increase in the cellular cAMP content in response to a beta-adrenergic agonist and the stimulation of membrane adenylate cyclase by the beta-agonist and/or GTP were markedly enhanced by the IAP treatment of C6 cells, but no change was induced in affinities of the agonist (or an antagonist) or GTP for their respective sites of action (or binding). The concentration of IAP required for the half-maximal enhancement was as low as 1 pg/ml, when the time of cell exposure to the toxin was prolonged to 18 h. No enhancement was observed for the basal cAMP content or basal enzyme activity, nor was activation of adenylate cyclase by Gpp(NH)p (or NaF) affected by IAP treatment. The Vmax value of a specific and low Km GTPase was significantly smaller in the membranes of IAP-treated cells than in those of control cells. Cholera toxin treatment of cells activated adenylate cyclase without exerting any influence on these IAP actions. Thus, IAP would appear to enhance beta-receptor-coupled stimulation of adenylate cyclase, in a manner distinct from cholera toxin, by rendering more GTP available to the GTP sites on the regulatory subunit of the receptor-enzyme system. PMID- 6277949 TI - The conformational transition of horse heart porphyrin c. AB - The heme iron of horse heart cytochrome c was selectively removed using anhydrous HF. The product, porphyrin c, exhibits the viscosity, far ultraviolet circular dichroic, and fluorescence properties characteristic for native cytochrome c. However, porphyrin c is more susceptible to denaturation by guanidine hydrochloride and by heat than is the parent cytochrome. All of the conformational parameters of porphyrin c exhibit a common reversible transition centered at 0.95 m guanidine hydrochloride at 23 degrees C and pH 7.0. Guanidine denatured porphyrin c refolds in two kinetic phases having time constants of 20 and 200 ms as detected by stopped flow absorbance or fluorescence measurement, with about 80% of the observed change in the faster phase. The kinetics of porphyrin c refolding are not significantly altered by increasing the viscosity of the refolding solvent 15-fold by addition of sucrose. We suggest that the folding of guanidine denatured cytochrome c is not a diffusion-limited process and that the requirement for protein axial ligation elicits the slow (s) kinetic phase observed in the refolding of cytochrome c. PMID- 6277950 TI - Kinetic effects of Ca2+ and Mg2+ on ATP hydrolysis by the purified ATP diphosphohydrolase. AB - ATP diphosphohydrolase (EC 3.6.1.5) catalyzes the hydrolysis of diphospho- and triphosphonucleosides and is sensitive to divalent cations. In this paper, we investigated the dependence of ATP hydrolysis on the concentration of free Mg2+ and Ca2+ and the cation ATP complexes. The enzyme was isolated from porcine zymogen granule membranes, solubilized in Triton X-100, and purified on a 5'-AMP Sepharose 4B affinity column resulting in a 1500-fold purification. Free unprotonated ATP4- was hydrolyzed in the presence of ethylene glycol bis(beta aminoethyl ether)-N,N,N',N'-tetraacetic acid. When hydrolysis rate was measured at different concentrations of the cation-ATP complex at constant free cation concentrations, normal hyperbolic curves were obtained. In CaCl2, both Kapp and Vapp increased as free Ca2+ increased from 25 to 1000 microM. In MgCl2, Kapp increased and Vapp decreased as free Mg2+ increased from 25 to 500 microM. From the rapid equilibrium rate equation, Ks and Vmax values of the substrates were calculated. We found that free ATP4-, Ca-ATP2-, and Mg-ATP2- are substrates and free cations do not bind the enzyme. PMID- 6277951 TI - Hormone-dependent changes in peroxisomal enzyme activity in guinea pig adrenal. PMID- 6277952 TI - Initiation, pausing, and termination of transcription in the threonine operon regulatory region of Escherichia coli. AB - The 6 S leader RNA transcript from the Escherichia coli threonine operon controlling region was synthesized in vitro using purified RNA polymerase and restriction fragment DNA templates. The terminated leader transcript was analyzed by RNase T1 digestion followed by electrophoresis on 20% polyacrylamide, 8 M urea gels. Oligonucleotides of 7, 8, 13, 15, and 35 bases in length were detected and correlated with the known DNA sequence. The kinetics of RNase T1 digestion indicated that the RNA forms extensive secondary structure, especially at the 3' terminus of the transcript. The sites of transcription initiation were determined by labeling the 5'-end of the transcript with [gamma-32P]ATP or -GTP followed by direct RNA sequencing. The DNA sequence preceding the initiation site shows homology with the equivalent regions of other bacterial and bacteriophage promoters. The transcription termination sites were determined by mapping of the RNase T1 oligonucleotides arising from the 3'-terminus of the transcript. Comparison of the mobilities of the 3'-oligonucleotides with the mobilities of standards on 20% polyacrylamide, 8 M urea gels indicated that the RNA contains a heterogeneous 3'-terminus. The two predominant oligonucleotides were CU7 and CU8. The 3'-terminus of the transcript also contains a region of dyad symmetry immediately preceding a stretch of uridine residues, characteristic of other rho independent transcripts. In addition, kinetic studies indicated that RNA polymerase pauses approximately 50 base pairs upstream from the site of termination. The pause site appears to be immediately distal to another region of dyad symmetry. PMID- 6277953 TI - Identification of a rat liver microsomal polypeptide involved in the transport of glucose 6-phosphate. Labeling with 4,4'-diisothiocyano-1,2-diphenyl[3H]ethane 2,2'-disulfonic acid. AB - Transport of glucose-6-P in intact rat liver microsomes is inhibited by 4,4' diisothiocyano-1,2-diphenyl[3H]ethane-2,2'-disulfonic acid ([3H]H2DIDS). The concentration of [3H]H2DIDS that inhibits transport activity by 50% is 35 microM. Glucose-6-P protected against the inhibition of transport activity caused by [3H]H2DIDS; mannose-6-P, 2-deoxyglucose-6-P, galactose-6-P, fructose-6-P, or glycerol-2-P did not. [3H]H2DIDS-treated microsomes were solubilized in sodium dodecyl sulfate and the microsomal polypeptides were separated by polyacrylamide gel electrophoresis. Labeled polypeptides were identified by autoradiography. Treatment of microsomes with concentrations of [3H]H2DIDS (50-100 microM) that resulted in maximal inhibition of transport activity allowed the identification of two microsomal polypeptides that contained most of the incorporated radioactivity. Their molecular weights were 54,000 and 59,000. The labeling of the former, but not the latter polypeptide was saturable and correlated linearly with the level of inhibition of transport activity. Concomitantly with its protective effect on translocase activity, the presence of glucose-6-P during the reaction with [3H]H2DIDS resulted in a significant increase in the amount of label incorporated into this peptide. This stimulation of labeling was specific for glucose-6-P. These results implicate the 54,000-dalton polypeptide as an obligatory component of the rat liver microsomal glucose-6-P translocase. PMID- 6277954 TI - A protein with multiple heme-binding sites from rabbit serum. AB - A 93,000 molecular weight protein (HBP.93) which binds hemin and protoporphyrin IX with high affinity has been isolated from rabbit serum using affinity chromatography on hemin-conjugated agarose. The amino acid composition of this protein is unique in that the proline and histidine contents are remarkably high (16.6 and 9.9 mol %, respectively). A large increase in the absorbance of the Soret region arises from the heme-protein interaction. The spectrophotometric titration showed that the protein can bind 25-35 mol of hemin/mol of protein. The apparent dissociation constant was estimated to be 1-4 X 10(-7) M for hemin at pH 7.4 and approximately 10(-6) M for protoporphyrin IX at pH 9.2. The similarity of the difference spectrum of heme-HBP.93 complex to that of heme-hemopexin complex suggests that a bisimidazol-type coordination of heme iron is involved in the binding. The extremely high capacity of HBP.93 to bind heme is also demonstrated by a large increase in the sedimentation velocity of the protein upon heme binding. The native heme-protein complex migrates faster than the heme-free protein in a polyacrylamide gel at pH 8.8; the increased mobility appears to be due to the charge on the carboxyl groups of the bound heme. Although the use of a hemin-agarose column has failed to reveal a protein of similar size and heme affinity in the sera of a number of other species, including man, the heme binding properties and high histidine level of the human alpha 2-histidine-rich glycoprotein raise the possibility that the two proteins are related. PMID- 6277955 TI - Isolation and properties of a cyclic AMP-binding protein from Neurospora. Evidence for its role as the regulatory subunit of cyclic AMP-dependent protein kinase. AB - A cyclic AMP-binding protein with a native molecular weight calculated to be 82,000 was purified 2,000-fold from Neurospora crassa. The apparent subunit molecular weight was 47,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, suggesting that the native protein exists as a dimer of identically sized subunits. The 8-N3-cyclic [32P]AMP-labeled protein appeared as a doublet upon two-dimensional gel electrophoresis, with pI = 5.4 and 5.5. Binding studies with both noncyclic and cyclic nucleotides showed cyclic AMP to have the highest binding affinity. The KD for cyclic AMP was 300 nM at 23 degrees C. The cyclic AMP-binding protein has two distinct cyclic AMP binding sites, a slowly dissociating site and a rapidly dissociating site. The cyclic AMP analog, 8-bromo cyclic AMP, was found to bind preferentially the slowly dissociating cyclic AMP binding site. The cyclic AMP-binding protein is thought to be the regulatory subunit of cyclic AMP-dependent protein kinase based upon the findings that it co-purifies with this protein kinase activity upon DEAE-cellulose chromatography, isoelectric focusing, and cyclic AMP affinity resin chromatography. On purification by the latter two procedures, no other cyclic AMP binding activity was detected. A number of similarities between the Neurospora cyclic AMP-binding protein and the cyclic AMP-dependent protein kinase regulatory subunits from yeast and mammalian tissue are discussed. PMID- 6277956 TI - Intra- and intermolecular strand transfer by HeLa DNA topoisomerase I. AB - The major type I DNA topoisomerase (topo I) has been purified from HeLa cell nuclei to a homogeneous, monomeric protein (Mr = 100,000). Similar to the nicking closing enzyme (Mr = 67,000) from rat liver (Been, M. D., and Champoux, J. J. (1981) Proc. Natl. Acad. Sci. U. S. A. 78, 2883-2887), HeLa topo I has the following properties: (a) HeLa topo I breaks down single-stranded DNA to smaller fragments, each with an enzyme-linked 3'-phosphoryl end and a free 5'-OH end. This cleavage is not dependent upon protein denaturant or protease treatment. (b) HeLa topo I produces single-stranded DNA circles from linear single-stranded DNA. Such DNA circles are believed to be produced by the intramolecular cyclization of topo I-linked, single-stranded DNA fragments. (c) HeLa topo I-linked, single stranded fragments (donors) can join covalently to double-stranded DNA possessing a 5'-OH group (acceptors). The donor is transferred to the 5'-OH end of the acceptor, independent of the position of the end (internal nick or end of linear DNA) or the configuration of the end (flush, 5'-protruding, or 5'-recessed end) of the acceptor. (d) HeLa topo I cleavage of single-stranded DNA is site specific, but no special sequence at the ends of the acceptor molecule is apparently required for a successful heterologous strand transfer. These results suggest that HeLa topo I may be involved in DNA sequence rearrangements in addition to its possible role as a swivelase for transcription and replication. PMID- 6277957 TI - The negative charge of articular cartilage surfaces. An electron microscopic study using cationized ferritin. PMID- 6277958 TI - Dyggve-Melchior-Clausen syndrome. A histochemical study of the growth plate. AB - The Dyggve-Melchior-Clausen syndrome is a rare inherited disorder in which a major feature, the skeletal dysplasia, is thought to result from a defect in endochondral ossification. To further characterize this disturbance, histochemical studies were performed on an iliac-crest biopsy specimen from a patient with disorder. The use of plastic embedding methods permitted previous observations, which were based on decalcified paraffin-embedded tissues, to be extended and a variety of new ones to be made. The resting cartilage matrix was very fibrous. In many areas it consisted of randomly oriented bundles of loosely woven fibers that stained as collagen. The chondrocytes in this region were excessively vacuolated and many contained cytoplasmic inclusions that stained non specifically as protein. Cartilage canals were prominent in the resting cartilage. In many areas, clusters of degenerating chondrocytes and occasional vacuolar lesions replaced the columns of proliferating and maturing cells that normally occupy the growth-plate region. There was coarse and irregular calcification of these clusters at the chondro-osseous junction, and intracartilaginous ossification was also found. CLINICAL RELEVANCE: These observations further define the histological criteria for the diagnosis of this disorder and provide new insights into its pathogenesis. PMID- 6277959 TI - Prostaglandin E2 and cyclic AMP in tumor and plasma of breast cancer patients. AB - Prostaglandin E2 and cyclic AMP (cAMP) levels were measured in tumors and plasma of 78 patients with benign and malignant breast tumors. Two groups of malignant tissues were found, one with a high level of PGE2 (M = 55.4 pg/mg) and one with a low level (M = 10.7 pg/mg). The low level did not differ significantly from the benign tissue level (M = 8.7 pg/mg). Two malignant groups could not be detected in the plasma levels. Plasma PGE2 concentration (in form of the 13,14-dihydro-15 Keto metabolite) did not reflect the tissue levels, and no difference was found between the benign (M = 59.9 pg/ml) and the malignant (M = 62.3 pg/ml) patients, but both concentrations were higher than those of healthy controls (M = 34.4 pg/ml). The stage of the cancer, the histological classification and, most important, the period of survival, could not be related to the differences in the PGE2 tissue levels. Neither could plasma cAMP be nominated as a breast cancer market because no difference was found between the cAMP levels of benign tumor patients (M = 16.48 pmol/ml), of malignant tumor patients (M = 21.14 pmol/ml) and of healthy controls (M = 19.07 pmol/ml). The conclusion is that although high amounts of PGE2 appear in some malignant breast tumors, they do not affect the clinical situation. These results may explain the failure to treat human breast cancer patients with prostaglandin synthetase inhibitors. PMID- 6277960 TI - Tissue connections in a transplantable virus-producing sebaceous adenoma of the mouse. II. A freeze-fracture study in conjunction with filipin. AB - The distribution, size, and configuration of intercellular junctions in the sebaceous tumor of mice were examined using the freeze-fracture technique. Three types of junctions were observed: desmosomes, gap, and tight junctions. Tight junctions in general consisted of short linear unbranched fibrils, and macular or complex tight junctional patterns were present only occasionally. Gap junctions ranged from small sports of 0.9 x 10(-3) micrometer2 to areas of approximately 0.46 micrometers2. Desmosomes were the most frequent junctional specializations, and it is concluded that they are at least partially responsible for this tumor not being metastatic. Filipin in conjunction with freeze-fracture showed filipin sterol complexes on the plasma membrane, the nuclear envelope, and the membranes of the endoplasmic reticulum and mitochondria. The intercellular junctions were devoid of these complexes indicating that these regions are low in cholesterol. PMID- 6277961 TI - Functional angiotensin II receptors in cultured vascular smooth muscle cells. AB - To study cellular mechanisms influencing vascular reactivity, vascular smooth muscle cells (VSMC) were obtained by enzymatic dissociation of the rat mesenteric artery, a highly reactive, resistance-type blood vessel, and established in primary culture. Cellular binding sites for the vasoconstrictor hormone angiotensin II (AII) were identified and characterized using the radioligand 125I angiotensin II. Freshly isolated VSMC, and VSMC maintained in primary culture for up to 3 wk, exhibited rapid, saturable, and specific 125I-AII binding similar to that seen with homogenates of the intact rat mesenteric artery. In 7-d primary cultures, Scatchard analysis indicated a single class of high-affinity binding sites with an equilibrium dissociation constant (Kd) of 2.8 +/- 0.2 nM and a total binding capacity of 81.5 +/- 5.0 fmol/mg protein (equivalent to 4.5 x 10(4) sites per cell). Angiotensin analogues and antagonists inhibited 125I-AII binding to cultured VSMC in a potency series similar to that observed for the vascular AII receptor in vivo. Nanomolar concentrations of native AII elicited a rapid, reversible, contractile response, in a variable proportion of cells, that was inhibited by pretreatment with the competitive antagonist Sar1,Ile8-AII. Transmission electron microscopy showed an apparent loss of thick (12-18 nm Diam) myofilaments and increased synthetic activity, but these manifestations of phenotypic modulation were not correlated with loss of 125I-AII binding sites or hormonal responsiveness. Primary cultures of enzymatically dissociated rat mesenteric artery VSMC thus may provide a useful in vitro system to study cellular mechanisms involved in receptor activation-response coupling, receptor regulation, and the maintenance of differentiation in vascular smooth muscle. PMID- 6277962 TI - Mannose-specific endocytosis receptor of alveolar macrophages: demonstration of two functionally distinct intracellular pools of receptor and their roles in receptor recycling. AB - Receptor-mediated endocytosis of rat preputial beta-glucuronidase and the glycoconjugate mannose-BSA by rat alveolar macrophages is inhibited by chloroquine and ammonium chloride. We have previously reported that these drugs cause a loss of cell surface binding activity and that they do not inhibit internalization of receptor ligand complexes when incubated with cells at 37 degrees C. In this report we more clearly delineate the intracellular site of weak base inhibition of receptor recycling and the mechanism of that inhibition. From our analysis of the kinetics of ligand transport we conclude that there are two functionally distinct intracellular pools of receptor. One of these, the cycling pool, is not sensitive to the presence of weak bases, and receptor-ligand complexes return from this pool to the cell surface intact. The second pool is responsible for the time-dependent intracellular delivery of ligand to acid vesicles, which is inhibited by weak bases. Chloroquine and ammonium chloride appear to inhibit the dissociation of receptor-ligand complexed in this second pool and thereby the production of free receptors for the continuation of receptor-mediated endocytosis. We examine the internalization and binding of ligand in normal and paraformaldehyde-treated cells and find that these are strongly affected by pH. In particular, the dissociation rate of receptor ligand complexes is enhanced greater than 7.5 fold by lowering the medium pH from 7 to 6. From these results we propose that weak bases raise the pH of acid intracellular compartments, slowing the rate of receptor-ligand dissociation and thereby reducing the cellular pool of free receptors available for further uptake of ligand. In addition, we demonstrate that receptor-ligand complexes cannot return to the cell surface from the amine-sensitive (acid) intracellular pool that led us to call this the nonreleasable pool. This final observation indicates that receptor movements through these two pools are functionally distinct processes. PMID- 6277963 TI - Isolation of bone cell clones with differences in growth, hormone responses, and extracellular matrix production. AB - Clones of nontransformed hormone-responsive bone cells have been isolated in vitro from mixed cell populations of fetal rat calvaria. In several independent isolations, microscopically visible colonies appeared at plating efficiencies of 5-10% of the starting cell numbers. Of these clones, approximately 10% grew to mass populations which could be assayed for a number of growth and biochemical properties. Although some similarities existed among the clones, they could be distinguished from each other and from the mixed cell populations. Population doubling times (tDs) and saturation densities varied over a wide range: e.g., tDs of 24-72 h and saturation densities of 0.4-5 x 10(5) cells/cm2. Morphologies varied from roughly polygonal multilayering cells to typically spindle-shaped monolayering cells. Hormone responsiveness, as measured by stimulation of cAMP by hormones, indicated that some clones were responsive to both parathyroid hormone (PTH) and prostaglandin E2 (PGE2), while others responded to PTH only. Analysis of extracellular matrix components revealed that all clones produced type I and type III collagens, though in different proportions. Similarly, although all clones synthesized four glycosaminoglycans (hyaluronic acid, heparan sulfate, chondroitin sulfate, and dermatan sulfate), the quantities of each were distinctive from clone to clone. Further investigation of such clones is continuing to define more precisely the heterogeneity of clonal bone cell populations in vitro. They represent an important step in the study of the endocrinology and differentiation of bone. PMID- 6277966 TI - Cat pial venoconstriction by topical microapplication of norepinephrine. PMID- 6277965 TI - Endoplasmic reticulum membrane isolated from small-intestinal epithelial cells: enzyme and protein components. AB - Endoplasmic reticulum membrane-rich fraction was obtained by subfractionation of the light microsomes from mouse jejunal mucosal epithelial cells. It was marked by high glucose-6-phosphatase, NADPH-cytochrome c reductase, and NADH-cytochrome c reductase activities and low Na+,K+-ATPase activity. The enrichment of Na+,K+ ATPase was 180-fold higher in the basolateral membranes than in the endoplasmic reticulum membrane-rich fraction relative to glucose-6-phosphatase. The protein peak that was phosphorylated in a Na-dependent manner was prominent in the basolateral membranes while it was a minor peak in the endoplasmic reticulum membrane-rich fraction. Under the electron microscope the fraction was seen to be composed of homogeneous small vesicles with thin smooth membranes. PMID- 6277967 TI - Cerebral histamine: indications for neuronal and vascular regulation. PMID- 6277964 TI - Cortical cell populations from rabbit kidney isolated by free-flow electrophoresis: characterization by measurement of hormone-sensitive adenylate cyclase. AB - Free-flow electrophoresis allows the separation of different cell populations from a cell suspension isolated from rabbit kidney cortex after perfusion of the kidneys with a calcium-binder, followed by gentle mechanical treatment. After electrophoretic separation, analysis of the adenylate cyclase activities after stimulation by various hormones allows the precise determination of the origin of the cell populations with different electrophoretic mobilities. Adenylate cyclase from the slow-moving main cell population was only sensitive to parathyroid hormone. These cells had also high alkaline phosphatase content, further demonstrating their proximal origin. The various fast-moving cell populations had adenylate cyclase sensitive to isoproterenol and arginine vasopressin but were less sensitive to parathyroid hormone than the slow-moving cells. Their alkaline phosphatase content was also much lower. This indicates that these fast-moving cell populations originate from both the granulous segment of the distal tubule and from the collecting ducts. The adenylate cyclase activity and the cyclic AMP contents of isolated proximal cells maintained in culture medium were also investigated. PMID- 6277968 TI - [Early gastric carcinoma. Clinicopathological analysis of 50 cases (author's transl)]. AB - 50 cases of early gastric carcinomas have been diagnosed in the past 7.5 years (1973 - June, 1980). The clinical pathological aspects of these cases are presented. Grossly, the depressed lesions (IIc, IIc + III), and histologically, the differentiated carcinoma composed the majority and accounted for 60% and 64% respectively. There were only 7 female patients in this series, and 5 were signet ring cell carcinomas. All 7 signet ring cell carcinomas (5 female, 2 males) were confined to the mucosal layer, although the largest one reached 4.5 X 3.4 cm. A small intestinal type early gastric carcinoma (1.2 X 0.8 cm), developing from severe atrophic gastritis was found after repeated follow-up examinations by gastroscopy in a three year period.U PMID- 6277969 TI - [Pelvic-peritoneal recurrences after amputation of the rectum for cancer. Report of 23 cases (author's transl)]. AB - Of 79 patients surviving after amputation of the rectum for cancer, local and regional recurrences occurred in 23 cases, in 3 out of 4 of these during the 2 year period following operation. Only 3 patients survived. This illustrates the frequency and serious nature of these recurrent lesions. Based on their experience, the authors consider the risk of recurrence to be doubled if one of the following factors is present : patient under 60 years of age, cancer located in the lower rectum, tumour over 4 cm in diameter, stages B or C according to Dukes' classification, an anaplastic or colloid structure. Enzyme assay and computed tomography are unfortunately not determining factors for early diagnosis and effective therapy of pelvic-peritoneal recurrences. the authors suggest, therefore, though it was never employed in their own series, that pre-operative irradiation could be of value, in those patients that have a high risk of developing local or regional recurrences. The general use of this technique is not suggested because its known inconveniences. PMID- 6277970 TI - Measurement of ketoconazole, a new antifungal agent, by high-performance liquid chromatography. PMID- 6277971 TI - Marijuana metabolites in urine of man. XI. Detection of unconjugated and conjugated delta 9-tetrahydrocannabinol-11-oic acid by thin-layer chromatography. AB - A method for separating unconjugated and conjugated delta 9-tetrahydrocannabinol 11-oic acid, the major urinary metabolite of delta 9-tetrahydrocannabinol in man, by liquid-liquid extraction and detection of both forms by thin-layer chromatography is described. The unconjugated form of the metabolite is extracted with hexane-diethylether (65:35), and the conjugated form (which remains in the aqueous phase) is extracted with ether after enzymic hydrolysis. The residue of each extract is chromatographed in an alkaline and an acidic solvent sequence, and the metabolites are detected with Fast Blue Salt B. PMID- 6277972 TI - Aromatic hydrocarbon metabolites in fish: automated extraction and high performance liquid chromatographic separation into conjugate and non-conjugate fractions. AB - An automated extractor-concentrator was used to extract metabolites of naphthalene, 2,6-dimethylnaphthalene, and benzo[a]pyrene from serum, bile and liver homogenate of rainbow trout (Salmo gairdneri). The extracts were analyzed by reversed-phase high-performance liquid chromatography (HPLC) with fluorescence detection. Recoveries of naphthalene and 2,6-dimethylnaphthalene metabolites from all matrices were generally greater than 90%; however, the recoveries of benzo[a]pyrene metabolites from serum ranged from 37-99%. In addition, conjugated metabolites of polycyclic aromatic hydrocarbons (PAHs) were separated from non conjugated metabolites and parent PAHs by using two diol columns with normal phase HPLC. The extraction and separation techniques were also applied to isolate metabolites in samples from fish fed 2,6-dimethylnaphthalene. PMID- 6277973 TI - A sensitive method for the production of diagnostic fingerprints of the genome segments of field isolates of rotavirus. AB - The 3'-terminal labelling procedure for analyzing the genome segment profile of field isolates of rotavirus (Clarke and McCrae, 1981, J. Virol. Methods 2, 203) has been further developed to produce a rapid and sensitive method for generating diagnostic fingerprints from individual species of double-stranded RNA. The fingerprints were obtained by a one-dimensional resolution of overlapping terminally labelled oligonucleotides produced by partial nuclease digestion with a base-specific nuclease. This fingerprinting method should be of great value in characterizing the molecular details of genome segment variation and will facilitate detailed molecular epidemiological studies of this important virus group. PMID- 6277974 TI - Detection of rotavirus immune complexes: relationship between rotavirus antibodies and rotavirus antigens in faeces. AB - An ELISA was developed for the identification of rotavirus immune complexes in pig faeces. The relationship between rotavirus antigens, rotavirus immune complexes and rotavirus antibodies of IgA class was examined. PMID- 6277975 TI - Immunoelectron microscopic study of the detection of the glycoproteins of influenza and Sendai viruses in infected cells by the immunoperoxidase method. AB - The envelope glycoproteins of influenza virus (HA and NA) and paramyxovirus (HN and F) were visualized on the surface of infected cells by immunoelectron microscopy using the indirect immunoperoxidase technique. In X7 influenza virus infected fibroblasts, the hemagglutinin (HA) and the neuraminidase (NA) were observed on the cell membrane respectively 2 and 3--4 h after infection. The antigens were initially seen as discrete patches and later evenly distributed along the plasma membrane prior to budding. Antibody induction of HA and NA was observed as cytoplasmic inclusions, with peroxidase-positive activity, attributed to endocytosis. The redistribution of HA and NA supports the hypothesis of lateral mobility of the viral glycoproteins in cellular membranes as visualized by the immunoperoxidase method. The glycoproteins of Sendai virus, in infected Madin--Darby bovine kidney cells, were found to be evenly distributed along the plasma membrane and endoplasmic reticulum, the latter by the indirect microperoxidase method. The immunoperoxidase methods may be useful for investigating the polarized distribution of envelope glycoproteins. PMID- 6277976 TI - Techniques for virus detection in aquatic sediments. AB - Laboratory experiments have been undertaken to study the adsorption-desorption of poliovirus to and from marine and freshwater sediments. It was observed that marine sediments retained 99% of added virus, whereas, freshwater sediments adsorbed approximately 40% of the added virus. Ten eluents were investigated for their ability to desorb viruses from a marine sediment. It was found that virus elution from the marine sediment was relatively low and ranged from less than 1% to 44%. Two eluents, urea-lysine and TCA-glycine at pH 9.0, were found to be the most efficient among the eluents tested. Viruses were more easily released from freshwater than from marine sediments. When urea-lysine, beef extract and purified casein were used as eluents, the overall virus recovery ranged from 8% to 22% for the marine sediment and from 23% to 59% for the freshwater sediments. The urea-lysine methods was used for the detection of indigenous enteroviruses in sediments from a closed shellfish bed. Enteroviruses were recovered with concentrations up to 708 TCID50/50 g of sediment. PMID- 6277978 TI - Comparison of solid-phase immune electron microscopy, direct electron microscopy and enzyme-linked immunosorbent assay for detection of rotavirus in faecal samples. AB - One hundred and fifteen faecal samples from children with gastroenteritis have been examined by solid-phase immune electron microscopy (SPIEM), standard direct electron microscopy and enzyme-linked immunosorbent assay (ELISA). Diluted high titered anti-(human) rotavirus serum, produced in rabbit by immunization with purified pooled rotaviruses from gastroenteritis patients, was used as antiserum in the immunological tests. The presence of rotavirus was demonstrated in 41 (36%) of the 115 samples by both SPIEM and ELISA. No false positive reactions were seen. By direct electron microscopy rotavirus was detected in 35 (30%) of the specimens. In addition, other gastroenteritis viruses like adeno-, astro- and calicivirus were visualized by this non-specific test in eight samples. The applicability of these three methods in routine diagnostic work is discussed. PMID- 6277977 TI - Enzyme-linked immunosorbent assay for acute adenovirus infection. AB - An enzyme-linked immunosorbent assay (ELISA) is described for demonstrating antibodies to the hexone antigen of adenoviruses. The antigen-coated, flat bottomed microtiter plates are incubated sequentially with dilutions of patients' sera (2 h at 37 degrees C) and peroxidase-coupled anti-human IgG (2 h at 37 degrees C). After a final washing, orthophenylenediamine is added to the plates, and the absorbance (A) measured 30 min later. The ELISA was found to be a hundred fold more sensitive than complement fixation. An evaluation methods for determining antibody concentration is described which correlates the absorbance of sera diluted 10(-3) to the absorbance of a reference serum containing an arbitrary value (100) of antibody. This methods avoids titration of sera and day to-day assay variations by different background reactions. A significant increase in antibody concentration of acute-phase serum over that of convalescent phase serum is observed. The ability to test sera in a single dilution and the automatic reading of results and their evaluation by computer make this assay suitable for diagnostic laboratories. PMID- 6277979 TI - Adrenal function in hirsutism. II. Effect of an oral contraceptive. PMID- 6277981 TI - Reduced Na+, K+ -ATPase activity in intact red cells and isolated membranes from obese man. AB - Na+, K+ -ATPase activity was measured in red blood cells from 20 nondiabetic euthyroid male Pima Indians with varying degrees of obesity; their body mass indices ranged from 22-60 kg/m2. The na+, K+ -ATPase, measured both by 86Rb uptake in intact cells and ATP hydrolysis by purified membranes, was inversely correlated with body mass index (r = -0.62; P less than 0.005 and r = -0.75; P less than 0.0001, respectively). These results confirm that obesity is associated with decreased Na+, K+ -ATPase in intact red blood cells, and provide the first demonstration of a reduced sodium pump in isolated red cell membrane preparations from obese men. PMID- 6277980 TI - The effect of prolactin on human aldosterone-producing adenomas in vitro. AB - There is evidence for an unidentified aldosterone-stimulating factor of pituitary origin. We measured the effect of ovine PRL (oPRL) on aldosterone secretion by isolated cell suspensions of human aldosterone-producing adenomas (APAs) and compared it to the effects of angiotensins, ACTH, and potassium (K+). In the first APA, the aldosteronotropic action of large doses of oPRL was double that of angiotensin II (AII); the response to ACTH was triple that to AII, while K+ had a small stimulatory effect. Results with the second APA showed that physiological concentrations of oPRL caused a response nearly double that to AII, but, once again, less than the response to ACTH; K+ was inert. ACTH contamination of the oPRL preparation was too minute to account for these findings. We conclude that oPRL possesses aldosterone-stimulating activity in APAs greater than that of angiotensins and potassium, but less that that of ACTH. These data suggest a role for PRL in aldosterone secretion by aldosterone-producing adenomas. PMID- 6277982 TI - Effect of naloxone on pituitary hypersecretory syndromes. PMID- 6277983 TI - Discrepancy between thyroid-stimulating and thyrotropin-binding inhibitory activities of Graves' immunoglobulin Gs assessed in the mouse. AB - TSH receptor binding-inhibiting (TBI) and thyroid-stimulating (TSI) activities of Graves' immunoglobulin Gs (IgGs) were measured using the murine TSH receptor (mTBI) and the McKenzie LATS bioassay. In contrast to the expectation that only the IgGs positive for LATS showed mTBI activity, mTBI activity was closely correlated with TBI activity measured at the human TSH receptor regardless of LATS potency (r = 0.79; P less than 0.01), indicating a lack of species specificity of IgG in this particular aspect. In 7 to 15 IgGs positive for both mTBI and LATS, there was a significant correlation between the 2 parameters (r = 0.89; P less than 0.01). However, 4 IgGs among the 15 were positive for mTBI but negative for LATS even when tested at 3- to 7-fold concentrations of IgG. PMID- 6277984 TI - Progressive adrenal failure in polyglandular autoimmune disease. AB - We describe the clinical course of a boy who developed progressive adrenal failure, beginning with failure of the zona glomerulosa, as part of polyglandular autoimmune disease. Initially the patient presented with hypoparathyroidism and mucocutaneous candidiasis. ACTH tests at ages 8 and 11 yr resulted in a normal response of both mineralo- and glucocorticoids. The constellation of hyponatremia , hyperkalemia, and growth failure at age 14 yr prompted a reevaluation. A repeat ACTH test, assessing individual contributions of zone fasciculata and glomerulosa, showed normal plasma cortisol, desoxycorticosterone, and corticosterone responses and a normal urinary response of 18 hydroxydeoxycorticosterone and tetrahydrodeoxycorticosterone. Urinary 18 hydroxycorticosterone and urinary as well as plasma aldosterone were undetectable. PRA was markedly elevated. The ACTH response of adrenal androgens, presumably metabolic products of the zona reticularis, was also deficient. Antiadrenal antibodies against all three layers of the adrenal cortex were present. Mineralocorticoid therapy resulted not only in normalization of electrolytes and PRA but also in catch-up growth. Repeat testing of fasciculata function at age 19 yr now shows that the patient's cortisol response to ACTH response in abnormal. The course of this patient suggest that in addition to monitoring the electrolyte status, periodic tests for both mineralo- and glucocorticoid synthesis should be performed in children with polyglandular autoimmune disease because progressive adrenal insufficiency may go unrecognized. PMID- 6277985 TI - Coupling defect of thyrotropin receptor and adenylate cyclase in a pseudohypoparathyroid patient. AB - A patient with type I pseudohypoparathyroidism was found to have mild hypothyroidism. The patient had an elevated basal TSH level and an exaggerated TSH response to TRH. There was no goiter despite increased TSH levels, and the 131I thyroidal uptake was low before and after exogenous TSH administration. These studies suggested that the patient might have partial resistance to TSH. The binding of radioiodinated TSH to thyroid membranes obtained by biopsy was next studied. The displacement of iodinated TSH by unlabeled TSH was found to be identical to that in normal control membranes. The adenylate cyclase stimulation by a supramaximal dose of TSH, however, was blunted (120.1 +/- 11.5 vs. 387.2 +/- 40.3 pmol cAMP/min/mg protein), while basal and NaF-stimulated activities were quite similar to the activities in normal membranes. These findings suggested a lack of signal transmission between the TSH receptor and the catalytic unit. Incubation of control membranes with TSH and GTP resulted in a synergistic effect on the adenylate cyclase activity. This was not found with the patient's membranes and suggested that the coupling failure was due to a defective guanine nucleotide regulatory protein. We conclude that in this case of type I pseudohypoparathyroidism, the associated mild primary hypothyroidism was due to a partial TSH refractoriness caused by a coupling defect between the TSH receptor and adenylate cyclase. This observation suggests that a common pathogenetic mechanism might underly type I pseudohypoparathyroidism and its associated hypothyroidism. PMID- 6277986 TI - End-organ response to adrenocorticotropin, thyrotropin, gonadotropin-releasing hormone, and glucagon in hypocalcemic magnesium deficient patients. AB - We have evaluated the responsiveness of hypocalcemic magnesium-deficient patients to ACTH, TRH, gonadotropin-releasing hormone, and glucagon as determined by the rise in serum cortisol, TSH, LH, and plasma cAMP concentrations, respectively. It was previously been shown that the hypocalcemia of magnesium deficiency is secondary to impaired secretion of parathyroid hormone (PTH) along with renal and skeletal resistance to the action of PTH. Since PTH secretion and action are though to be effected through the intermediary action of cAMP, and magnesium is a required cofactor for adenylate cyclase, defective generation of cAMP could account for the observed defects in PTH secretion and action. Other hormonal systems requiring the intermediary action of cAMP may be similarly affected by magnesium deficiency. The results of the present study, however, demonstrate normal responsiveness of the adrenal cortex, thyrotrophs, gonadotrophs, and liver to their respective trophic hormones in hypocalcemic magnesium-deficient patients. The reason why these responses are intact while PTH secretion and action are impaired is unknown but may be accounted for by differing magnesium requirements of the adenylate cyclase complex in these tissues. PMID- 6277987 TI - Distribution of guaiacol peroxidase in human endometrium and endocervical epithelium during the menstrual cycle. AB - Guaiacol peroxidase (G-Px) was measured in extracts from five sections along the length of human uterus on different days of the menstrual cycle or after menopause. The lower uterine-endocervical region had a significantly higher G-Px content (expressed as enzyme units per g wet tissue) than the other sections, although in postmenopausal patients the G-Px activity was uniformly low in all sections of the uterine cavity. We observed no significant changes in G-Px levels during the menstrual cycle, except, possibly, a decrease around ovulation, which precluded a positive correlation between plasma estrogen levels and uterine G-Px content; such estrogen dependence of G-Px has been previously shown in the rat. In vitro, G-Px was inhibited by estriol and 17 beta-estradiol, marginally inhibited by estrone, and most notably inhibited by the catecholestrogens tested (2-hydroxy-17 beta-estradiol, 2-hydroxy-estriol, and 2-hydroxy-estrone), which were equipotent inhibitors; LH and FSH, progesterone, or cortisol had no effect on G-Px activity. We hypothesize that catecholestrogens are natural substrates and regulations of G-Px activity in the human uterus. PMID- 6277988 TI - Marrow transplantation for leukemia. AB - Marrow transplantation for selected patients with leukemia, as for patients with severe combined immunologic deficiency or severe aplastic anemia, has now become an accepted clinical procedure. For patients with acute leukemia who have relapsed after achieving a remission of chemotherapy, marrow grafting from an identical twin or an HLA-identical sibling has now been demonstrated to produce median remissions as long as or longer than any reported for combination chemotherapy. In contrast to chemotherapy, marrow transplantation offers the possibility of cure for a small but significant fraction of these patients. Marrow transplantation for patients with ANL in first remission has now resulted in median survivals much longer than any reported with chemotherapy. Although it now appears that more than 50% of these patients can be cured with marrow transplantation, a much longer follow-up is indicated since some patients who achieve a complete remission with combination chemotherapy are now living for a long time, and some of these patients (less than 20%) may also be cured. Current intensive research with new modalities such as interferon, Acyclovir, Cyclosporin A, and monoclonal antibodies can reasonably be expected to improve the overall results of marrow transplantation. PMID- 6277989 TI - Toxigenic Clostridium perfringens from a parvovirus-infected dog. AB - A strain of Clostridium perfringens, type A, has been isolated from the intestine of a dog which died from parvovirus infection. This Clostridium strain produces a toxin which can be detected by counterimmunoelectrophoresis, using C. difficile antitoxin, and produces cytotoxicity in WI-38 cell culture. Cytopathology can be blocked by C. difficile antitoxin. Its role in canine parvovirus infection is unknown. PMID- 6277990 TI - Band 1 and voltage-sensitive Na+ channels are not codistributed in cultured rodent neuronal cells. AB - The relationship of the mouse nervous system specific band 1 protein to the putative high molecular weight component of the Na+ channel was investigated using antibody to band 1. Morphologic differentiation of cultured neuroblastoma cells has been reported to increase the quantity of the putative Na+ channel high molecular weight component. Morphologically differentiated clone NB2a neuroblastoma cells have 2-3 times the amount of band 1 and 1.5 times the relative rate of synthesis of band 1 as undifferentiated cells. The anti-band 1 serum reacts with both adult mouse and rat brain but not 3 cultured rat neuronal lines known to have active Na+ channels. Thus either band 1 is not a component of the Na+ channel or individual cultured murine neuronal lines has distinct macromolecular Na+ channels. PMID- 6277991 TI - Pathogenesis of paralysis and lymphoma associated with a wild mouse retrovirus infection. Part 1. Age- and dose-related effects in susceptible laboratory mice. AB - Wild mouse ecotropic retrovirus (Cas-Br-M) induced paralysis and non-thymic lymphomas in susceptible NIH Swiss and NFS/N mice. The incidence of paralysis was highest and latency shortest in mice receiving high doses of virus. Lower dose inoculation and inoculation of older mice produced less paralysis with longer latency, but resulted in more lymphomas. However, 10-day-old mice did not develop paralysis and had fewer lymphomas. Anti-Cas-Br-M antibody was detectable in sera from 10-day-old infected mice but not from paralyzed mice. These data suggest that while paralysis and lymphoma may result from different virus-host interactions, the development of immunocompetence may play a role in the age dependent resistance to Cas-Br-M-associated paralysis and lymphoma in these mice. PMID- 6277992 TI - Characterization of opiate-mediated responses of the feline ileum and ileocecal sphincter. AB - Although opioid peptides have been demonstrated immunohistochemically in the feline intestine, the action of these peptides is unknown. The aims of this study were: (a) to determine the distal ileal and ileocecal sphincter (ICS) responses to morphine sulfate (MS), methionine enkephalin (ME) and leucine enkephalin (LE); (b) to determine the mechanism by which exogenous opiates mediate these responses; (c) to determine the type of receptor involved in mediating these responses and (d) to ascertain whether endogenous opiate-mediated responses may be vagally induced. The ICS responded to all three opiate agonists with tonic and phasic contractions, the latter being associated with increased spike activity. The ED(max) for ICS pressure response was 1 mug/kg for ME, 5 mug/kg for LE, and 150 mug/kg for MS. The distal ileum responded with increased spike activity and phasic contractions. The ED(max) for the ileal motility index response was 1.0 x 10(-1) mug/kg for ME, 1 mug/kg for LE, and 150 mug/kg for MS. Thus, both sites demonstrated similar dose-response relationships, both responding to at least 100 times lower doses of enkephalins than MS. The ICS contraction preceded ileal contractions. The ileal and ICS response was not antagonized by atropine, hexamethonium, phentolamine, propranolol, cinanserin, or tetrodotoxin. Naloxone, 600 mug/kg, antagonized the response to the enkephalins while 10 mug/kg antagonized the response to MS. Higher doses of the specific-receptor agonist SKF 10047 and kappa-receptor agonist ketocyclazocine were required before a contractile response was elicited. Electrical stimulation of the cervical vagus induced ICS contraction and a fall in blood pressure. The ICS contractile response but not the blood pressure response was inhibited by naloxone 1 mg/kg. These data indicate: (a) tonic and phasic ICS contraction followed by ileal contraction may be mediated through delta-type opiate receptors located in the muscle membrane and (b) opiate-mediated ICS contraction may be induced during vagal stimulation. PMID- 6277994 TI - A possible biological role of the 'biologically inactive' region of polypeptide hormones. PMID- 6277993 TI - Role of angiotensin-converting enzyme in Bacille Calmette-Guerin-induced granulomatous inflammation. Increased angiotensin-converting enzyme levels in lung lavage and suppression of inflammation with captopril. AB - Lung lavage levels of angiotensin-converting enzyme (ACE)-like activity were increased in C57BL/6 mice with Bacille Calmette-Guerin (BCG)-induced chronic granulomatous pulmonary inflammation and splenomegaly. Contrariwise, ACE activity was not increased in lung lavage fluids of CBA mice that developed only minimal pulmonary inflammation in response to BCG. ACE-like activity correlated with the intensity of inflammation and Captopril, a specific competitive inhibitor of ACE activity, markedly suppressed the induction and maintenance of the BCG-induced inflammatory response in both lungs and spleen. It was necessary, however, to provide sustained treatment with large doses of Captopril in order to reduce the inflammatory response. After a single intraperitoneal injection of Captopril, ACE levels in lung lavage of BCG-injected mice were reduced but returned to preinjection levels or greater within 24 h. The highest dose of Captopril was more effective in reducing the lung fluid level of ACE in BCG-inflamed lungs. This suggests that sustained daily injections of Captopril were necessary to maintain reduced ACE levels. In vitro studies indicated that high concentrations of Captopril did not affect macrophage mobility or chemotactic activity for macrophages. Thus, ACE may act as a molecular mediator of BCG-induced granulomatous inflammation in the lung. PMID- 6277995 TI - Comparison of ELISA, SPACE, and electron microscopy for the routine diagnosis of rotavirus infection. AB - Previous studies on the serological diagnosis of rotavirus infection have utilised locally produced antibodies. In this study we have compared two commercially produced assays, an ELISA (Rotazyme, Abbott) and a newly developed assay--solid phase aggregation of coupled erythrocytes (SPACE) (Wellcome Research Laboratories), with electron microscopy (EM). The SPACE test appeared less sensitive than EM. The ELISA was shown to be as sensitive as EM but more versatile. Our experience suggests that the ELISA could be successfully incorporated into the routine of any diagnostic laboratory. PMID- 6277996 TI - Phase I study of ceftizoxime, a new cephalosporin. Repeated dosing study. PMID- 6277997 TI - Comparison of imipramine- and nortriptyline-induced orthostatic hypotension: a meaningful difference. PMID- 6277998 TI - Identification of and interactions between noradrenergic and serotonergic neurites in the myenteric plexus. AB - Interactions between enteric noradrenergic and serotonergic neurites in the myenteric plexus were examined. The influence of exogenous norepinephrine (NE) and endogenously released NE on the release of 3H-serotonin (3H-5-HT) from electrically stimulated, everted segments of guinea pig small intestine were analyzed. In addition, methods were employed to permit the ultrastructural identification of enteric serotonergic and noradrenergic neurites. These included electron microscopic radioautographic localization of 3H-5-HT in gut from animals treated with 6-hydroxydopamine (6-HD; 100 mg/kg) and examination of NaMnO4-fixed tissue from animals given desmethylimipramine and the indolic neurotoxin, 5,7 dihydroxytryptamine (5,7-DHT). Exogenous NE antagonized the stimulated release of 3H-5-HT; this action was mediated through alpha adrenoceptors. Evidence was obtained, however, that endogenous NE liberated from sympathetic postganglionic nerve terminals had a different action on the stimulated release of 3H-5-HT from that of exogenous NE. Endogenous NE appeared to facilitate 3H-5-HT release through an action on beta adrenoceptors. Terminals identified as probably serotonergic by radioautographic labeling with 3H-5-HT or by loading with 5,7-DHT were most often found to end on neuronal somata or proximal dendrites. Synaptic specializations were found by radioautography in these locations. Terminals, dendrites, and cell bodies that took up 5,7-DHT could be identified in NaMnO4 fixed material that also permitted the simultaneous recognition of noradrenergic varicosities. Apparent noradrenergic-serotonergic axoaxonic contacts were found by means of this double-labeling technique. Together with the results of the experiments on the release of 3H-5-HT, these anatomical observations suggest that noradrenergic axons form facilitatory axoaxonic synapses with enteric serotonergic neurons. PMID- 6277999 TI - Ketoconazole in griseofulvin-resistant dermatophytosis. AB - The efficacy of ketoconazole was evaluated in twenty patients with chronic dermatophyte infections who had failed to clear with griseofulvin therapy. Trichophyton rubrum was the causative organism in nineteen of the patients, and Trichophyton mentagrophytes in one patient. Three of twelve organisms tested showed in vitro resistance to griseofulvin. Duration of infection ranged from 2 to 28 years. Patients received 200 to 400 mg of ketoconazole daily for periods up to 8 months. In addition, patients were followed for 5 months post-therapy to monitor recurrences. Clearing was seen clinically as early as 2 weeks, and by 18 weeks all patients showed marked improvement or clinical clearing, though only six achieved complete mycologic cure. Improvement followed a predictable sequence of sites, with lesions of the trunk healing first, followed by hands, feet, and finally, nails. After 8 months, though all patients showed proximal nail clearing, onychomycosis persisted in thirteen of twenty affected sites. By 5 months post-therapy, four of six patients who had achieved clearing of skin and nails showed recurrences. No significant side effects were observed during therapy, though rare, apparently idiosyncratic cases of hepatotoxicity have been reported. Ketoconazole is an affective therapeutic agent for griseofulvin resistant dermatophytosis. Apparent cures may subsequently recur after discontinuation of therapy. PMID- 6278000 TI - Adenoma sebaceum of Pringle: a clinicopathologic review, with a discussion of related pathologic entities. AB - "Adenoma sebaceum of Pringle" (ASP) is a misnomer. The tumor is not an adenoma and is not derived from sebaceous glands. The lesion is characterized by dermal fibrosis and associated vascular proliferation and dilatation. Changes in contiguous sebaceous glands and other adnexal structures are merely secondary. Thus, "angiofibroma" would be a more appropriate name. The histologic changes in ASP (and in related pathologic lesions) suggest that it is a hamartoma rather than a true neoplasm. However, the embryologic tissue of its origin is not definitively known. PMID- 6278001 TI - Melanin pigmentation of the skin in primary biliary cirrhosis. AB - A histological and ultrastructural study has demonstrated that cutaneous pigmentation in primary biliary cirrhosis (PBC) is due to the presence of increased amounts of melanin, widely dispersed throughout both epidermis and dermis. No deposits of stainable iron were observed. Compared with skin from matched sites from control patients with alcoholic cirrhosis and no pigmentation, the melanocyte: keratinocyte ratio was not significantly higher in PBC. However, in PBC, melanosomes persisted to unusually high levels in the epidermis and were packaged in larger membrane-bound clusters than was the case in the controls. Whether excess melanin results from increased melanogenesis or defective melanin degradation remains unclear, although there is some evidence favouring the latter mechanism. No hormonal (beta-MSH and ACTH) or chemical (bile salt irritation) stimuli to increase melanogenesis were demonstrated. PMID- 6278002 TI - Large potentiation of agonist response in intact cells is produced by increases only in GTP-dependent adenylate cyclase activity. AB - Treatment of cultured SV40-transformed normal rat kidney cells with the drug, 2 pyridine carboxylic acid, results in a pronounced potentiation in the ability of isoproterenol, prostaglandin E1, and cholera toxin to elevate cyclic AMP levels. With isoproterenol, the initial rate of cyclic AMP accumulation and the maximum cyclic AMP attainable are increased, and also the time of maximum cyclic AMP is prolonged. GTP-dependent adenylate cyclase activities are potentiated in crude membranes from the treated cells, but no evidence for alterations in cyclic nucleotide phosphodiesterase or release of cyclic AMP into the medium could be demonstrated. Results show that augmented adenylate cyclase activity alone, without changes in phosphodiesterase, can lead to dramatic alterations in cyclic AMP accumulation in response to cyclase agonists. PMID- 6278003 TI - Discovery and recognition of calmodulin: a personal account. PMID- 6278004 TI - A GTP-protein activator of phosphodiesterase which forms in response to bleached rhodopsin. AB - A specific protein associated with rod-outer-segment disc membranes binds GTP only in the presence of bleached rhodopsin. Once formed the protein-GTP complex becomes a soluble activator of cGMP phosphodiesterase. It is shown that this activator complex can be completely separated from rhodopsin and retain its ability to activate phosphodiesterase when added to a pool of totally dark (unilluminated) disc membranes. The photoreactive GTP analogue p3-(4 azidoanilido)-5' GTP (AAGTP) is shown to be a more effective substrate than GTP, Gpp(NH)p or 8-azido GTP. [8, 5' 3H] AAGTP was used to specifically covalently label the GTP-binding protein. The protein labeled exhibits a mass of 40,000 daltons when analyzed by SDS-PAGE. PMID- 6278005 TI - Forskolin: a unique diterpene activator of cyclic AMP-generating systems. AB - Forskolin, a diterpene of the labdane family, activates adenylate cyclase in broken cell preparations as well as in intact tissues. This activation does not require the guanine nucleotide regulatory subunit of the enzyme and probably occurs via an interaction with the catalytic subunit of adenylate cyclase. Activation of adenylate cyclase by forskolin results in marked increases in levels of intracellular cyclic AMP in a variety of eukaryotic cells. Low concentrations of forskolin which alone elicit small increases in intracellular cyclic AMP greatly potentiate hormonal activation of adenylate cyclase in a number of intact cells. Forskolin elicits cellular responses which have been proposed to be dependent o cyclic AMP as a second messenger. Forskolin, thus provides an invaluable tool for the investigation of the role of cyclic AMP in physiological responses to hormones, both through it direct activation of adenylate cyclase and through its ability to potentiate hormonal activation of adenylate cyclase. PMID- 6278006 TI - Cyclic nucleotides and the control of epithelial cell proliferation: cyclic CMP may be a partial mediator of the response of the pigeon crop-sac to prolactin. AB - The possible role of cyclic nucleotides as second messengers mediating hormone induced cellular growth in vivo was investigated using the proliferative response of the pigeon crop-sac mucosal epithelium to prolactin (PRL) as a model system. Local injections of cAMP, cCMP, cGMP, cTMP or cUMP alone over the prolactin responsive cells had no mitogenic effect. When injected along with a small dose of PRL, cAMP at doses above 1 micromole inhibited the response by 10%. While cGMP at a dose of 10 micromoles augmented the response by 47%. In fact, cCMP gave a log-linear dose-response relationship with significant augmentation of the response to PRL observed with doses as low as 0.01 micromole. Cyclic UMP, cTMP and various 5'-nucleotide monophosphates had no effect on the response to the hormone. When the dibutyryl analogs of the cyclic nucleotides were tested for their ability to potentiate the response to PRL, only dibutyryl cCMP was effective. These data suggest that cCMP may be a partial mediator of the proliferative action of PRL in this system, but it is probably not the sole second messenger for prolactin's action in these cells. Our results also indicate that failure of a cyclic nucleotide to mimic a hormone-induced response by itself does not prove that the compound is uninvolved in the hormones's action. PMID- 6278007 TI - A sensitive equilibrium binding assay for soluble beta-adrenergic receptors. AB - An equilibrium binding assay has been developed for digitonin-solubilized beta adrenergic receptors using 125 I-pindolol (IPIN) as a radioligand. Up to 50% of the beta-adrenergic receptors from rat lung membranes could be solubilized using 1% digitonin. Following incubation of soluble fractions with IPIN at 25 degree, protein associated radioactivity was identified by column chromatography using Sephadex G-50. The solubilized receptors bound IPIN with properties similar but not identical to those of the membrane bound receptor. The Kd determined for IPIN binding to soluble receptors was 113 pM while the Kd for membrane associated receptors was 36 pM. The rate constant for association (k1) of IPIN was 0.15x10(9) M-1 for soluble receptors and 2.2x10(9) M-1 min-1 for lung membrane receptors. The rate constant for dissociation (k2) was 0.025 min-1 for soluble receptors and 0.048 min-1 for membrane receptors. Agonists and antagonist of beta adrenergic receptors inhibited in a stereoselective manner the binding of IPIN to both soluble and membrane bound receptors. The affinities of individual drugs determined for soluble receptors were similar to those determined for membrane receptors. Not only could digitonin-solubilized receptors be identified in soluble preparations from rat lung, but also from rat cerebral cortex and liver, and from L6 muscle, C6 rat glioma, and 1321N1 astrocytoma cell membranes. PMID- 6278008 TI - A new principle for resistance to cholera: desensitization to cyclic AMP-mediated diarrhea induced by cholera toxin in the mouse intestine. AB - The mechanisms behind the intestinal resistance to cholera toxin was studied in a mouse model. Repeated peroral treatments with cholera toxin (CT) led to a long lasting inhibition of the toxin-induced activation of intestinal adenylate cyclase (AC). A corresponding inhibition of the intestinal fluid secretion induced not only by CT but also by prostaglandin E1 was observed. This unspecific desensitization was followed by a CT-specific inhibition of secretion and AC after 8 to 16 days. The desensitization to CT was totally reversed by a 4 hour treatment with cycloheximide, an inhibitor of protein synthesis. Neither the secretory response to dibutyryl-cyclic AMP nor the activity of soluble phosphodiesterase differed between the CT-treated mice and the control group. Nor was the average turn-over rate of intestinal cells changed as judged from the mucosal incorporation of [3H]-thymidine. It is postulated that intestinal resistance to CT is mainly a function of AC-desensitization mediated by an inducible protein. PMID- 6278009 TI - Cautions on the use of the heat stable inhibitor of protein kinase: studies with S49 lymphoma cells. AB - When crude preparations of protein kinase inhibitor (PKI) are added to homogenates of S49 lymphoma cells, protein kinase activity increases rather than decreases. This is not an increase in the activity of cyclic AMP-dependent protein kinase, which is completely inhibited by PKI, nor an increase in either cyclic GMP-dependent or Ca ++-dependent kinases. The increased kinase activity is not due to PKI itself but to one or more protein contaminants which serve as substrates for an S49 cell kinase which is cyclic nucleotide- and Ca ++ independent. These contaminating substrates can be readily separated from PKI by DEAE cellulose chromatography. Although the crude PKI preparations sold by the major biochemical supply houses may be satisfactory for some purposes, we suggest that crude PKI be further purified when used to assess kinase activity in systems as complex as homogenates of whole cells. Otherwise, the presence of contaminating proteins in crude preparations of PKI can lead to erroneous conclusions about the type and quantity of protein kinase in the cell. PMID- 6278010 TI - Rapidity of cortisol transfer between blood and milk following adrenocorticotropin injection. AB - Cortisol concentrations in milk and blood plasma were measured in 12 lactating Holstein cows following administration of 40 IU adrenocorticotropin intravenously (groups 1 and 2) or 4 ml saline (control). Blood and milk samples were collected at 15 or 30-min intervals for 4 h from control and group 1 cows and at hourly intervals for 4 h from group 2. Cortisol concentrations in plasma and milk were increased 15 min after treatment and peaked by 1 h in group 1 but remained unchanged in controls. Group 2 cows were sampled less frequently but showed a pattern for plasma and milk cortisol concentrations similar to group 1. The profile of increased cortisol concentrations and of their decline 1 to 2 h later was similar for plasma and milk. These results suggest that cortisol concentrations in milk reliably indicate cortisol concentrations in blood and may be valuable in monitoring stress in dairy cows, depending upon duration of stress and its timing relative to milk removal. PMID- 6278011 TI - [Growth and fatty acid composition of Clostridium perfringens cells under constant pH values]. PMID- 6278012 TI - Dietary fiber consumption in an adult Dutch population. AB - The intake of dietary fiber in an adult Dutch population aged 25 to 65 years was found to be 24.0 +/- 6.9 gm. per day (mean +/- SD), or 10.5 +/- 2.6 gm. per 1,000 kcal. Important sources of dietary fiber were bread and other cereals (32 percent of dialy intake), potatoes (17 percent), other vegetables (24 percent), and fruit (15 percent). For estimating group means of daily dietary fiber intake, equal precision can obtained by using a seven-day record method or by collecting one day records from twice as many subjects. PMID- 6278013 TI - Use of spin label and the flow-induced ESR spectral difference for studying erythrocyte deformation. AB - ESR spin-labeling method is expanded to measure the macroscopic visco-elastic properties of erythrocytes. A suspension of erythrocytes with an incorporated fatty acid spin label was forced to flow through a flat ESR sample cells, and the ESR spectral change caused by the shear flow was utilized to assess the cell deformability. Chemical cross-linking or heat denaturation of membrane proteins to make the cells less deformable without any morphological change was found to reduce the relative spectral difference (delta h/h). This result indicates that the spectral difference is related to the cell deformation that accompanies the orientation of the cells in the shear flow. In addition, the average decay time (tau av) for the spectral difference observed when the flow was abruptly interrupted became shorter with an increase in the degree of cross-linking or heat-denaturation abruptly interrupted became shorter with an increase in the degree of cross-linking or heat-denaturation at 49 degrees C. Since the observed tau av is much shorter than the expected rotational correlation time for the erythrocyte, the decay is attributed to the deformation recovery process. It is demonstrated that the measurements of both delta h/h and tau av by ESR spectroscopy give qualitative information on the viscosity and the elasticity of the cell membrane system. PMID- 6278014 TI - Beta-lipotropin and beta-endorphin in physiological and surgical menopause. AB - Beta-lipotropin (beta-LPH) and beta-endorphin (beta-EP) plasma levels were characterized by a significant decrease in postmenopausal females (22 subjects aged from 56 to 70 yr) when compared to fertile women (22 subjects from 31 to 45 yr). On the contrary, ACTH plasma levels determined in 10 of the premenopausal and 13 of the postmenopausal subjects reported above showed constant levels in both groups. A significant increase in the beta-LPH/beta-EP molar ratio was observed in postmenopausal females. The plasma beta-LPH and beta-EP levels studied before and 6 months after ovariectomy, showed a significant decrease in 8 out of 10 patients, while they remained constant in the other 2. Two subjects, in whom postsurgical samples were taken during a flushing episode, showed beta-LPH and beta EP plasma levels which were both higher than the corresponding preovariectomy values. The results suggest that these changes may be important in explaining modifications in behavior and mood frequently found in postmenopausal females and in patients undergoing surgical castration in the fertile age. PMID- 6278015 TI - Pituitary gigantism in a 31 month old girl: endocrine studies and successful response to hypophysectomy. AB - A case of pituitary gigantism occurring in a 31 month old female child is reported. Growth records indicate that the disorder began early in the second yr of life. Apart from her size and history of excessive sweating, there were no characteristic clinical features of endocrinopathy. Elevated and autonomous secretion of GH (60-109 microgram/l) and prolactin were corrected by the removal of an eosinophilic pituitary adenoma. In the subsequent 6 yr, despite the presence of immunoreactive GH (4.6-17.3 microgram/l), plasma somatomedin was subnormal and the patient showed growth failure which responded normally to exogenous GH therapy. This case, which appears to be the youngest example of verified pituitary gigantism on record, illustrates that a successful outcome can be achieved by surgical ablative therapy. PMID- 6278016 TI - Measurement of blood volume in fetal and neonatal sheep using red blood cells labelled with 99m technetium. AB - Blood volume has been measured in fetal and neonatal sheep using red blood cells labelled with 99mTc. The calculated volumes were highly correlated with simultaneous measurements made using the standard 51Cr labelled red cell method, although in absolute terms the 99m Tc method provided volumes which on average exceeded by a small percentage those determined with the 51Cr method. Measurements using the 99mTc method were also made at different ages in fetal and neonatal sheep and, while no correlation could be demonstrated between blood volume and either fetal or neonatal age, neonatal blood volumes were highly correlated with body weight. The 99mTc method is considered to be a reliable technique for measuring perinatal blood volumes in sheep with the short half-life of the isotope offering additional advantages. PMID- 6278017 TI - An outbreak of gastroenteritis caused by both rotavirus and Shigella sonnei in a private school in Rio de Janeiro. AB - In May 1980 an extensive outbreak of gastroenteritis occurred in a private school in the city of Rio de Janeiro. Examination of faeces and paired sera showed that this outbreak was caused by both rotavirus and a virulent strain of Shigella sonnei. In the first 19 stool samples collected seven (37%) had rotavirus only, six (32%) had Sh. sonnei only, while four (21%) had both agents. Examination of the second and third stool collections revealed only the presence of Sh. sonnei. The 18 paired sera showed seroconversion for rotavirus in four cases (22%) and in seven cases (39%) for Sh. sonnei. The overall attack rate of the disease was approximately 75%, the nursery and kindergarten having higher attack rates. Students in all grades became sick at the same time, and the unimodal curve of the onset dates of symptoms indicates a common source outbreak. Evidence suggested a contaminated water supply. PMID- 6278018 TI - An outbreak of infectious hepatitis investigated using radioimmunoassays for hepatitis A virus antibody. AB - An outbreak of suspected infectious hepatitis in a primary school was investigated using sensitive diagnostic methods for hepatitis A. A total of 116 sera from children were tested for the presence of both IgM and IgG antibodies to hepatitis A (HAV). The results were compared to those obtained for samples from a control school. IgG antibodies were present in 45% and 10% of the children in the outbreak and control schools respectively. PMID- 6278019 TI - Antibody to cytomegalovirus in Malta. AB - Antibody to cytomegalovirus (CMV) was sought in sera from Malta using immunofluorescence. Seven per cent of the infants, 36% of the school children, increasing to 100% of the adults aged over 40 years were found to have antibody. Most infection occurred in pre-school children and adults over 25 years of age. This pattern of antibody acquisition appears different from that described for other countries. PMID- 6278020 TI - Chemotactic factor inactivation by stimulated human neutrophils mediated by myeloperoxidase-catalyzed methionine oxidation. AB - Leukocyte chemoattractants were inactivated when exposed to human neutrophils and either ingestible particles or phorbol esters. Loss of biologic activity was time and temperature-dependent, required physiologic concentrations of viable neutrophils and a halide, and was inhibited by azide or catalase. Neutrophils from patients with either hereditary myeloperoxidase deficiency or chronic granulomatous disease failed to inactivate the chemoattractants unless purified myeloperoxidase or H2O2, respectively, was added. Susceptibility to inactivation by neutrophils correlated with the presence of methionine in the attractant. Loss of chemotactic activity was blocked by low concentrations of methionine and by higher concentrations of other reducing agents, but was unaffected by oxidized methionine. Paper chromatography demonstrated that exposure of a formyl-methionyl peptide chemotactic factor to either the cellfree myeloperoxidase system or stimulated neutrophils resulted in its conversion to a molecular species whose location in the chromatographs was identical to that of the peptide containing oxidized methionine. Thus, stimulated human neutrophils inactivate peptide chemoattractants by secretion of myeloperoxidase and H2O2, which combine with halides to form oxidants that react with a critical methionine residue. We suggest that myeloperoxidase-catalyzed oxidation of thioethers may constitute an inflammatory control mechanism as well as a general means of modifying the functional properties of biologic mediators. PMID- 6278021 TI - Monocyte aggregation and superoxide anion release in response to formyl-methionyl leucyl-phenylalanine (FMLP) and platelet-activating factor (PAF). AB - Like the PMN, human peripheral blood monocytes were capable of aggregating in response to FMLP and PAF. Monocyte aggregation was dependent on glycolysis and the presence of divalent cations. Unlike the PMN, monocyte aggregation in response to FMLP was not accompanied by degranulation, nor was it potentiated by cytochalasin B. Furthermore, cytochalasin B and its cogenitor, dihydrocytochalasin B, inhibited aggregation in response to PAF. PAF and FMLP appeared to react with the monocyte at separate receptors because sequential challenge of the monocyte with the same agents failed to elicit further aggregation, whereas rechallenge with the alternative agent induced further aggregation. Because the monocyte will aggregate to chemotactic agents in vitro, it is likely that the cell will be affected by these agents in vivo, which could lead to leukoembolization of circulating monocytes. PMID- 6278022 TI - Lipopolysaccharide receptor on rabbit peritoneal macrophages. I. Binding characteristics. AB - The Bordetella pertussis endotoxin, labeled with tritium ((3H)-LPS), bound irreversibly and nonspecifically to rabbit lung macrophages, but bound reversibly and specifically to both resident and elicited rabbit peritoneal macrophages. The specific binding capacity of the macrophages was saturated with about 3 X 10(4) LPS molecules per cell. The binding was inhibited with the homologous unlabeled endotoxin, but not at all with endotoxin from Proteus mirabilis, thus assessing ligand specificity. Endotoxins from other bacteria gave intermediate inhibition value. Binding of tritium-labeled pertussis endotoxin was significantly inhibited by one of the two polysaccharides (PS-1) present in this endotoxin, but neither the other polysaccharide (PS-2) nor the Lipid A fragment exhibited such activity. These results strongly suggest the presence of a lectin-like receptor for LPS on the membrane of rabbit peritoneal macrophages. PMID- 6278023 TI - Selective inhibition of immunoregulatory cell functions by cyclosporin A. PMID- 6278024 TI - Expression on normal lymphocytes of two cell surface antigens, XenCSA and GIX, related to the major glycoproteins (gp70) of murine leukemia viruses. AB - XenCSA and GIX are two cell surface antigens related to the major envelope glycoproteins (gp70) of murine leukemia viruses. The levels of expression of these gp70 determinants were assessed in 36 recombinant inbred mouse strains and selected backcrosses derived from crosses between C57BL/6 with DBA/2 and C3H/He. These two antigens segregated in backcross mice and showed a different strain distribution pattern among the recombinant inbred mice, demonstrating that XenCSA and GIX are distinct genetic markers for different endogenous gp70 sequences. It was also shown that independent sets of gene regulate the expression of XenCSA and GIX. PMID- 6278025 TI - Generation of cytotoxic lymphocytes by SV40-induced antigens. AB - In order to study the correlation of in vivo tumor transplantation immunity and in vitro immunologic assays, cell-mediated cytotoxicity against SV40-transformed cells was studied in AL/N strain mice by using 51Cr-release assay. Killing of SV40-transformed AL/N fibroblast cells was observed by spleen cells of AL/N mice immunized with syngeneic SV40-transformed cells. Immunization with the solubilized SV40 tumor-specific transplantation antigen (TSTA) that induced transplantation immunity in vivo did not elicit cytotoxic spleen cells in vitro. However, the spleen cells from mice immunized with solubilized TSTA and then sensitized in vitro with SV40-transformed cells became cytotoxic against SV40 transformed fibroblasts. Similarly, SV40 TSTA (T antigen) purified by immunoprecipitation was able to prime the lymphocytes in AL/N mice: the primed lymphocytes could differentiate into cytotoxic lymphocytes upon in vitro stimulation by SV40-transformed cells. These data indicate that SV40 TSTA (T antigen) plays a role in the induction of cytotoxic lymphocytes. PMID- 6278026 TI - Detection of H-2 and Sendai virus antigens by chemiluminescence. AB - Two monoclonal antibodies reacting with Ia9 and H2.2, respectively, have been added to spleen cell suspensions prepared from mice of different H-2 haplotypes. Cellular light emission was monitored in a liquid scintillation spectrometer operated in the out-of-coincidence mode. The antibodies stimulated chemiluminescence (CL) in cells possessing the target antigenic determinant, but were inactive in cells lacking the determinant. In addition, CL could be stimulated in Sendai virus-infected spleen cells with anti-Sendai antibodies. The results demonstrate that CL measurement is a sensitive and simple method for the detection of cell surface antigens and for the screening of antibodies reacting with these antigens. PMID- 6278028 TI - The role of temperature and swab materials in the recovery of herpes simplex virus from lesions. PMID- 6278027 TI - New attenuation marker for junin virus based on immunologic responses of guinea pigs. AB - A new attenuation marker to distinguish a virulent strain (XJJV) from an attenuated strain (XJC13JV or XJOJV) of Junin virus by means of the humoral and cellular responses to unrelated antigens was studied in guinea pigs. Strain XJJV suppressed the humoral immune response, as shown by the lower titers of precipitating antibody to ovalbumin. The concomitant decrease in serum complement level contributed to a milder Arthus cutaneous reactivity. In contrast, the attenuated strains did not decrease the humoral response. The pathogenic strain suppressed cell-mediated immunity, as demonstrated by decreased contact sensitivity to 2,4-dinitro-1-fluorobenzene and by depression of delayed skin reactions to tuberculin purified protein derivative. When attenuated strains were used, such suppressive effects were not observed. For virulent strain XJJV, virus replication in lymphoid organs and immunosuppressive effects were correlated. These findings provide a further means to differentiate between virulent and attenuated strains of Junin virus for the purpose of vaccine control of Argentine hemorrhagic fever. PMID- 6278029 TI - Concept review of genital herpes vaccines. PMID- 6278030 TI - [Biochemical aspects of arrhythmia (author's transl)]. PMID- 6278032 TI - [C-AMP concentration in various organs of female rats and in human ovaries with aging (author's transl)]. AB - The change in 3',5'-cyclic adenosine monophosphate (c-AMP) concentration was observed in various organs of rats in gonadal cycle in adult group and with aging (30, 70, 100, 120 weeks), and in human ovaries with aging. 1) The average c-AMP concentration of ovaries of rats showed a significant change with estrus cycle and was higher in the following sequence: proestrus, diestrus II, diestrus I and estrus phase. This tendency was also seen in hypothalamus and pituitary, but was not statistically significant, 2) The average c-AMP concentration in tissues began to decline significantly from 70 weeks in cerebral cortex and hypothalamus, and from 80 weeks in ovaries. However, on the other hand the concentrations in pituitary, liver and adrenal declined markedly from 100 weeks. 3) The c-AMP in ovaries of 80 weeks rats by pregnant mare serum (PMS) road increased by 0.5-fold in concentration, and by 0.6-fold in whole tissue relative to that of 30 weeks rats. 4) A significant difference in serum LH and FSH level between ovarian artery and vein was not found in cycling mature group, non-cycling climacteric group and post-menopausal group of women. 5) Both average concentrations and total values of c-AMP in ovaries of non-cycling climacteric and post-menopausal women were lower than those of mature cycling women. This fact may imply a different response by ovarian tissues such as corpus luteum, follicle and other tissues to gonadotropin. From these results of c-AMP in tissues, it is concluded that the decline of ovarian function with aging of rats was relatively earlier than pituitary, although being delayed compared with hypothalamus, and were the ovarian function in humans declined in the premenopausal period. PMID- 6278033 TI - [Studies on the interaction of prolactin and estrogen in rabbit ovary and uterus (author's transl)]. AB - The biological effects of prolactin (PRL) on the uterus and ovary, and the cooperative action of PRL and estrogen to these tissues were investigated in the female white rabbit. 1) Estrogen receptor (ER), progesterone receptor (PR, not in ovary) and PRL receptor were demonstrated in the rabbit uterus and ovary. The dissociation constant determined by Scatchard analysis was approximately at the level from 10(-10)M to 10(-9)M in each ligand binding to receptor. 2) In the pseudopregnant rabbit, b-PRL (30 iu/kg) increased the uterine and ovarian weight (p less than 0.001, p less than 0.01 respectively) but neither altered the steroid levels, nor the uterine and ovarian cytoplasmic ER levels. In the uterus, b-PRL increased the PRL receptor levels and tended to increase the nuclear ER levels. 3) In the ovariectomized immature rabbit, b-PRL (30 iu/kg) increased in uterine weight (p less than 0.05), decreased the uterine cytoplasmic ER levels (p less than 0.05), but increased the nuclear ER levels (p less than 0.05). With estrogen, b-PRL increased the uterine weight (p less than 0.05); tended to increase the cytoplasmic and nuclear ER levels, and increased the cytoplasmic PR levels (p less than 0.05). 4) In the ovariectomized immature rabbit, b-PRL did not change the histological finding of the uterus, whether estrogen was primed or not. In the pseudopregnant rabbit, b-PRL accelerated the progestational proliferation of the uterus. These results indicated that PRL demonstrated the biological effect on the rabbit uterus and ovary directly via PRL receptor, and that PRL enhances the estrogenic action, giving the effect on the mechanisms related with ER in the rabbit uterus. PMID- 6278031 TI - [(2'-5') oligoadenylate and its synthetase (author's transl)]. PMID- 6278034 TI - [The studies for exacerbating factors of Cushing's syndrome during pregnancy, report of a case (author's transl)]. PMID- 6278035 TI - A concise review: common viral exanthems and enanthems. PMID- 6278036 TI - Abdominal scintigraphy: locating the site of intestinal bleeding. PMID- 6278037 TI - Lithium prevents saline deoxycorticosterone acetate (DOCA) hypertension in the rat. AB - Uni-nephrectomized rats drinking 1% saline instead of water, were given Doca intramuscularly, 50 mg/kg BW per week for 2 weeks. The mean blood pressure in the control group was 105 +/- 4 (+/- S.E.) mm Hg, whereas in the Doca-saline group it rose to 152 +/- 5.1 (p less than 0.001). Rats, similarly treated, were placed on daily intraperitoneal lithium injections of either of two doses: 1.5 or 3.0 mEq/kg BW per day. Their blood pressures, at the end of 2 weeks of treatment, were 117 +/- 2 mm Hg and 103 +/- 2.1, respectively (p less than 0.001 vs. lithium untreated Doca-saline rats). Water-drinking rats, receiving daily intraperitoneal lithium (3 mEq/kg BW) for 2 weeks, had normal blood pressures, not different from the controls (104 +/- 11 mm Hg). The Doca-saline and Doca-saline-lithium (1.5 mEq/kg/day) groups had similar changes in mean daily body weight, plasma sodium, osmolality, and GFR. The renal beta-adrenergic receptor density in the Doca saline-lithium rats was in the normal range, 51 +/- 4.5 mol/mg of protein. In the Doca-saline hypertensive rats, it was significantly lower, being 27.2 +/- 3 fmol/mg of protein, p less than 0.05. These renal plasma cell membrane beta adrenergic receptors were characterized by direct tissue binding with ( )[3H]dihydroalprenolol. These results show that lithium prevents the development of hypertension in the Doca-saline rats. The effect of lithium on the sympathetic nervous system is a possible mechanism in the prevention of the Doca-saline hypertension. PMID- 6278038 TI - Alterations in renal tubular water transport induced by parathyroid hormone: evidence for both antidiuretic hormone-mediated and independent effects. PMID- 6278040 TI - Carcinoma in a calcified pleomorphic adenoma of the submandibular gland. PMID- 6278039 TI - Characterization of hereditary partial myeloperoxidase deficiency. AB - We studied a family with a partial myeloperoxidase deficiency. The myeloperoxidase in the neutrophils and monocytes of the parents and their two sons had normal spectral properties (determined optically and by EPR). Enzymic characteristics (oxidation of iodide) were indistinguishable from those of normal myeloperoxidase; moreover, immunological identity between the myeloperoxidase in the leukocytes of the family members and normal myeloperoxidase was found. No differences in heat stability were observed. The neutrophils and monocytes of the sons contained 9% to 18% of the myeloperoxidase content of normal cells; the neutrophils and monocytes of the parents contained 45% to 58%. These data suggest either that the parents are heterozygous and the sons homozygous for hereditary partial myeloperoxidase deficiency or that each parent is heterozygous for a different type of myeloperoxidase deficiency and the sons combine both deficiencies. The oxidative metabolism of the neutrophils during phagocytosis was not affected by the myeloperoxidase deficiency. The killing of Staphylococcus aureus was apparently normal. The perforation of Escherichia coli by the neutrophils of the sons, however, was retarded in comparison with normal neutrophils. PMID- 6278041 TI - Human testis does not secrete oestrone sulphate. AB - The concentrations of five steroids in samples of spermatic venous blood collected from 17 men undergoing ligation of varicocoeles were compared with those in samples from the antecubital vein. There was evidence of testicular secretion of testosterone, androstenedione, oestradiol-17 beta and oestrone, since the ratios of the mean concentrations in spermatic venous plasma to those in peripheral venous plasma were 77.2, 9.1, 28.7 and 1.6 respectively. The testicular secretion of oestrone sulphate was minimal; the ratio of the mean concentrations in spermatic and peripheral plasma was 1.07. These results support the view derived from isotope dilution studies that almost all oestrone and oestrone sulphate in the circulation is derived from peripheral conversion of other precursor steroids. PMID- 6278042 TI - Effects of hydroxysteroid dehydrogenase inhibitors on in-vitro and in-vivo steroidogenesis in the ovine adrenal gland. AB - The effectiveness of trilostane and azastene as inhibitors of adrenal steroidogenesis was compared by in-vitro and in-vivo methods. A radioimmunoassay was developed for the measurement of cortisol in ovine plasma, incubation medium and tissue extract using a specific antiserum raised against cortisol 21 acetate,3-carboxymethyloxime:bovine serum albumin. Trilostane (20 mumol/l) decreased cortisol synthesis and release both in unstimulated and in ACTH stimulated adrenal tissues in vitro. The same concentration of azastene had a lesser effect on unstimulated adrenals and was completely ineffective in blocking the stimulatory action of ACTH. In vivo, trilostane suppressed adrenal steroidogenesis in pregnant and cyclic ewes but the suppression in pregnant ewes was over a longer period, and after lower doses. It is concluded that trilostane had an inhibitory effect on ovine adrenal steroidogenesis both in vitro and in vivo. PMID- 6278043 TI - An experimental study of the relation of cardiac jelly to the shape of the early chick embryonic heart. AB - The structural roles of cardiac jelly components were examined in the early developing chick embryonic heart. Cardiac jelly matrix components were enzymically removed in situ by injecting nanogram quantities of enzymes directly into the cardiac jelly. Injection of ovine testicular hyaluronidase caused shrinkage and the heart became flaccid, but overall heart shape did not change. These responses were the result of enzymatic removal of glycosaminoglycan sugar moieties and were not due to lumenal collapse. Although purified collagenase did not cause any noticeable change, enzymes with non-specific proteolytic activity induced marked cardiac shape changes. In such hearts the dorsal mesocardium opened completely, and the myocardium as well as splanchnic mesoderm of foregut detached from their substrate and the entire heart region swelled. Consequently the shape of the heart was altered completely. The results suggested that in the normal condition the myocardial envelope was under an internal pressure due to the presence of glycosaminoglycans in the cardiac jelly space, and that some matrical non-collagenous protein components were essential to control the internal pressure. Therefore it is suggested that the internal pressure of cardiac jelly may be the direct driving force for the looping process and protein components of cardiac jelly may be important in directing the force for the morphogenetic process. PMID- 6278044 TI - Growth factors and pattern formation. AB - Growth and pattern formation occur simultaneously in many epimorphic fields and it has been suggested that specification of positional information is somehow linked to cell division. It is possible, therefore, that boundary regions responsible for the specification of positional information produce cell growth factors. In this paper I review the properties of some known growth factors, describe their effects on the cell cycle and discuss how they might act. In developing a convenient in vitro assay for morphogenetic factors it will be much easier in measure incorporation of [3H]thymidine into responding cells than to estimate changes in positional value. PMID- 6278045 TI - Membrane properties of neuroglia in the optic nerve of Necturus. AB - The optic nerve of Necturus has proved a useful preparation for the study of glial cell membranes in vivo and in vitro with anatomical relations to axons intact and isolated following axon degeneration. The glial membrane potential behaves as a selective potassium diffusion potential; there is no evidence of a significant permeability to other naturally occurring ions. The specific membrane resistance of the glial cells is high compared to that of neurones; there are low resistance intercellular connexions among the cells which permit the passage of both ions and the dye Lucifer Yellow. The cells are readily and reversibly uncoupled by procedures which decrease the intracellular pH. There is no evidence for voltage-sensitive sodium channels in the membrane. Following sodium gain and potassium loss the membrane displays a potassium-dependent strophanthidin sensitive electrogenic sodium pump. The glial membrane is depolarized by potassium released from active axons as well as by glutamate. The glial depolarization contributes to potentials recorded with surface electrodes. Depolarization by K+ plays a role in the redistribution of K+ which locally accumulates around active neurones and also affects glial metabolism and glucose uptake. PMID- 6278046 TI - K+ changes in the extracellular space of the spinal cord and their physiological role. AB - K+ accumulates in the intercellular space as a result of neuronal activity. The changes in extracellular K+ concentration, delta[K]e (estimated by K+-selective microelectrodes), depends on neuronal activity, on the density of discharging neurones and the removal of the accumulated K+ by diffusion, active transport and current flow through cells. In the mammalian as well as the amphibian spinal cord a single volley in a peripheral nerve increases [K]e by 0.2-0.5 mmol . 1-1, while tetanic stimulation (100 Hz) by 7-8 m-mol . 1-1, with a maximum in the lower dorsal horn. Increased [K]e was also found in lumbar segments when the somatosensory cortex of the cat and medulla of the frog were stimulated. In the frog spinal cord, the tactile stimulation of the hindlimb evoked delta[K]e by about 0.1 mumol . 1-1, nociceptive stimulation by 0.2-1.0 mmol . 1-1. Spontaneous delta[K]e and dorsal root potentials (DRPs) were observed at various intervals after stimulation, associated with the decay phase of delta[K]e. It was shown that primary afferent depolarization (PAD) consists of two components: the 'early' component (mediated by GABA and depressed by picrotoxin or bicuculline) and the 'late' K+ component (potentiated by picrotoxin and bicuculline). Even when increased [K]e produces PAD, this does not mean that it also results in presynaptic inhibition. It was found that the delta[K]e produced depolarization of motoneurones and neuroglia and there is every reason to believe that this also applies to the interneurones. Evidence is available that an increase of [K]e up to 6 mmol . 1-1 facilitates impulse transmission in the spinal cord while higher levels result in its inhibition. PMID- 6278047 TI - J chain is encoded by a single gene unlinked to other immunoglobulin structural genes. AB - Immunoglobulin J chain mediates the polymerization of both IgM and IgA immunoglobulins. Its synthesis is closely regulated in B lymphocytes, apparently at the level of RNA transcription. To define the genetic bases of this regulation, we have determined the location and number of J chain genes in the mouse. Analysis of DNA from a group of somatic cell hybrids containing various mouse chromosomes on a constant background of Chinese hamster chromosomes indicated that this gene is located on mouse chromosome 5, unlinked to immunoglobulin heavy and light chain structural genes. Restriction mapping experiments further suggested the existence of a single J chain gene per haploid genome. This result was confirmed by quantitative analyses of band intensities yielded by Southern blots of mouse genomic DNA and J gene-containing plasmid DNA. PMID- 6278048 TI - Evidence for the horizontal acquisition of murine AKR virogenes by recent horizontal infection of the germ line. AB - Several recent reports (8, 10, 11, 13) have established the biological and molecular genetic similarity between the endogenous AKV virus of strain AKR, and an N-ecotropic endogenous virus found in the genome of feral Japanese mice, Mus musculus molossinus. The similarities are so striking as to suggest a common origin of these viruses, which are present in some, but not all, inbred mouse strains. The virogenes of AKR mice may have been acquired by either: (a) common descent of AKR (and other AKV(+) strains) from a common ancestor of AKR and molossinus animals, or (b) horizontal germ line infection of the AKR strains by molossinus virus at 1;he strain's inception followed by fixation through inbreeding. The sexual descent model carries with it a prediction of relative consanguinity of the AKR strain and molossinus, whereas the horizontal infection model does not. We have examined the polymorphic allozyme (allelic isozyme) genotype of 51 nonvirus-related loci in 17 strains of mice including AKR, C58, BALB/c, Swiss, and molossinus. By comparing the composite allozyme genotype of different inbred and outbred mouse strains, the "genetic distance" statistic was derived. Genetic distance measures the degree of allelic substitution between populations and increases proportionately with the amount of time the populations have been reproductively isolated. The genetic distance computed between molossinus and AKR is large, nearly 5-10 times the distance between known related populations and strains (e.g., C57L vs. C57BL/6). Molossinus had a similarly large distance from AKV negative strains (Swiss, C57L) as it did from AKV- positive strains. Cellular DNA sequences that flank the integrated AKV provirus were analyzed by restriction enzyme digestion of liver DNA from molossinus, AKR, and additional inbred strains that express ecotropic murine leukemia virus. The integration flanks of three AKR provirus sequences, Akv-1, Akv-2, and a third uncharacterized sequence, were not evident in molossinus cell DNA, which contained at least six different proviral integration fragments. These data effectively exclude the interpretation of consanguinity of AKR and molossinus and support the notion of acquisition of the endogenous virus in AKR by horizontal infection of the molossinus virus. PMID- 6278049 TI - Allelism and linkage studies of murine leukemia virus activation genes in low leukemic strains of mice. AB - Previously, we identified two genes, termed Inc-1 and Inb-1, that interact to enhance ecotropic murine leukemia virus induction in low virus strains of mice. Mice related to BALB/c in origin carry a locus termed Inc-1, whereas mice related to B6 carry an Inb-1 locus. Mice that carry both Inc-1 and Inb-1 yield 10- to 50 fold more virus-producing cells than parental strains on induction with halogenated pyrimidines in vitro and demonstrate enhanced murine leukemia virus production in vivo. Here, we show that mice related to BALB/c in origin, i.e., A, C3H/He, and SEC, have an Inc-1 locus that is allelic with that of BALB/c. The C57BR mouse strain has an Inb-1 locus that is allelic with that of B6, located on chromosome 8, 30 cM from Es-1. We also show that the Inc-1 locus of BALB/c mice is located on chromosome 5, 24 cM from Pgm-1 and 43 cM from Gus. Kozak and Rowe (6,8) and Ihle and co-workers (3) have shown that the ecotropic virus-inducing genes in BALB/c and B10 mice are located on chromosomes 5 and 8, respectively, with similar distances from the previously mentioned biochemical markers. Our data are consistent with two possibilities: Inc-1 and Inb-1 are part of the virus inducing genes Cv-1 and Bv-1, respectively , or Inc-1 and Inb2-1 are tightly linked regulatory genes. PMID- 6278051 TI - A three-barrier model for the hemocyanin channel. AB - Keyhole limpet hemocyanin forms ion-conducting channels in planar lipid bilayer membranes. Ionic current through the open hemocyanin channel presents the following characteristics: (a) it is carried mainly by cations; (b) it is a nonlinear function of membrane potential; (c) channel conductance is a saturating function of ion activity; (d) it shows ionic competition. A model for the open hemocyanin channel is developed from absolute reaction rate theory. The model calls for three energy barriers in the channel. Two energy barriers represent the entrance and exit of the ion into and out of the channel. The third barrier separates two energy minima that represent two binding sites. Furthermore, only one ion is allowed inside the channel at a given time. This model is able to recreate all the hemocyanin characteristics found experimentally in negatively charged and neutral membranes. PMID- 6278050 TI - Block of endplate channels by permeant cations in frog skeletal muscle. AB - Motor endplates of frog semitendinosus muscles were studied under voltage clamp. Current fluctuations induced by iontophoretic application of acetylcholine were analyzed to give the elementary conductance, gamma , and mean open time, tau , of endplate channels. Total replacement of the external Na+ ion by several other metal ions and by many permeant organic cations changed both gamma and tau . Except with NH4+ ions, the gamma values with foreign test ions were all smaller than expected from the independence relation and their previously measured permeability ratios. The more hydrophobic ions gave the smallest gamma values. Foreign permeant cations also depress gamma when mixed with Na+ ions. These effects could be interpreted in terms of binding of ions to a saturable site within the endplate channel as they pass through. The site for organic ions would have a hydrophobic component. Similar evidence is given for a metal ion binding site on the cytoplasmic end of the channel accessible to internal ions. Most foreign cations also shortened tau when applied externally. The changes of gating did not seem to be correlated with changes in gamma . Thus there is no evidence for control of tau by ions bound within the pore. PMID- 6278052 TI - The "late" Ca channel in squid axons. AB - Squid giant axons were injected with aequorin and then treated with seawater containing 50 mM Ca and 100-465 mM K+. Measurements of light production suggested a phasic entry of Ca as well as an enhanced steady-state aequorin glow. After a test K+ depolarization, the aequorin-injected axon was stimulated for 30 min in Li seawater that was Ca-free, a procedure known to reduce [Na]i to about one-half the normal concentration. Reapplication of the elevated K+ test solution now showed that the Ca entry was virtually abolished by this stimulation in Li. A subsequent stimulation of the axon in Na seawater for 30 min resulted in recovery of the response to depolarization by high K+ noted in a normal fresh axon. In axons first tested for a high K+ response and then stimulated in Na seawater for 30 min (where [Na]i increases approximately 30%), there was approximately eight fold enhancement in this response to a test polarization. Axons depolarized with 465 mM K seawater in the absence of external Ca for several minutes were still capable of producing a large phasic entry of Ca when [Ca]0 was made 50 mM, which suggests that it is Ca entry itself rather than membrane depolarization that produced inactivation. Responses to stimulation at 60 pulses/s in Na seawater containing 50 mM Ca are at best only 5% of those measured with high K solutions. The response to repetitive stimulation is not measurable if [Ca]o is made 1 mM, whereas the response to steady depolarization is scarcely affected. PMID- 6278054 TI - RNA polymerase binding sites on the broad host range plasmid RP4. AB - Binding sites of Escherichia coli RNA polymerase on RP4 plasmid DNA were determined electron microscopically. Comparison of the RNA polymerase binding map and the genetic map of RP4 revealed several strong binding sites outside the well known RP4 genes. RNA polymerase binding sites for the three antibiotic resistance genes were also detected. Two binding sites were observed for the tra-1 region, whereas the tra-2 and tra-3 regions showed no prominent affinity for RNA polymerase. The genomic regions for the replication origins, oriV (for vegetative replication) and oriT (for transfer replication, equivalent to rlx), both exhibited strong binding to RNA polymerase, as did genomic regions which code for trans-acting replication functions (trfA and trfB). PMID- 6278053 TI - Analyses of bipolar cell responses elicited by polarization of horizontal cells. AB - Simultaneous intracellular recordings were made from a bipolar cell and a horizontal cell in the carp retina. The properties of the bipolar cell were studied while injecting current into the horizontal cell. Hyperpolarization of horizontal cells, irrespective of their type, elicited a hyperpolarizing response in on-center bipolar cells and a depolarizing response in off-center bipolar cells. Analyses of the ionic mechanisms of bipolar cell responses revealed that depolarization of horizontal cells simulated and hyperpolarization opposed the effect of central illumination. The effect of polarization was exerted in such a manner that each type of horizontal cells modified the transmission from those photoreceptors from which they receive main inputs. In on-center bipolar cells, for example, the L-type horizontal cells receiving inputs mainly from red cones modified the cone-bipolar transmission accompanied by a conductance change of K+ and/or Cl- channels, and the intermediate horizontal cells receiving inputs from rods modified the rod-bipolar transmission accompanied by a conductance change of Na+ channels. In off-center bipolar cells, the effect of polarization of any type of horizontal cells was mediated mainly by conductance changes of Na+ channels. Feedback mechanisms from horizontal cells to photoreceptors could explain these results reasonably well. PMID- 6278055 TI - Extraction of spore-lytic enzyme from Clostridium perfringens spores. AB - Various chemical reagents known to extract spore coat protein were used to extract spore-lytic enzyme (SLE) from intact and germinated spores of Clostridium perfringens. Of the reagents tested, 7.2 M-urea plus 10% (v/v) mercaptoethanol, pH 2.85, solubilized the most SLE activity per mg spores. The quantity of SLE extracted was dependent on the initial pH of the reagent, with a maximum between pH 2.7 and 3.0. Germinated spores yielded more SLE than non-germinated spores upon urea/mercaptoethanol extraction. SLE release during spore germination probably utilizes a trigger mechanism not satisfied by germination alone. Significant amounts of SLE were released during germination when spores were suspended in potassium chloride or a complex germinant mixture containing brain heart infusion, yeast extract and chloramphenicol, but not during germination with sodium nitrite, which non-enzymically lysed the cortical peptidoglycan. Greater solubilization of SLE activity was obtained by urea/mercaptoethanol extraction of spores germinated with nitrite than of spores germinated with either potassium chloride or the complex germinant. PMID- 6278056 TI - Reduced expression of Tn 10-mediated tetracycline resistance in Escherichia coli containing more than one copy of the transposon. AB - In Escherichia coli a single copy of Tn10 confers high-level resistance to tetracycline. Resistance itself results from expression of three distinct mechanisms which normally act together (Shales et al., 1980). In cells containing two copies of Tn10, the level of resistance to tetracycline was reduced. This was not due to overproduction of the repressor which controls the resistance genes, because strains diploid for an operator-constitutive allele of Tn10 also exhibited reduced expression of resistance. The negative gene dosage effect resulted from decreased expression of two mechanisms (1 and 2) consequent on enhanced expression of the third mechanism. The net result of increasing the copy number was a decrease in resistance because mechanism 3 was less efficient than mechanisms 1 and 2 in protecting the cell against tetracycline. The DNA sequence responsible for the reduced expression of resistance was contained in the internal BglII fragment of Tn10. This sequence, which is probably unique to Tn10, may encode the protein which mediates mechanism 3. Elevated levels of this protein probably cause decreased expression of mechanisms 1 and 2. PMID- 6278057 TI - Comparison of thymidine kinase activities indiced in cells productively infected with herpesvirus saimiri and herpes simplex virus. AB - The replication of unselected strains of herpesvirus saimiri (HVS) was sensitive to bromodeoxyuridine and bromovinyldeoxyuridine (BVdU) but insensitive to acycloguanosine (ACG), in contrast to the growth of herpes simplex virus (HSV) which was sensitive to all three analogues. Mutants of HVS resistant to bromodeoxyuridine and BVdU could be selected by growth in the presence of these inhibitors. Productive infections of owl monkey kidney or Vero cell cultures by unselected strains of HVS resulted in increases in a thymidine kinase (TK) activity which was deficient in cells infected with bromodeoxyuridine-resistant mutants of the virus. Induction of the virus enzyme promoted a net increase in the uptake and incorporation of exogenous labelled thymidine in the face of the progressive inhibition of the overall incorporation of [35S]methionine and [3H]uridine into productively infected cells. The TK induced in cells infected with HVS differed from the major activity of uninfected cells and resembled that encoded by HSV in its capacity to phosphorylate iododeoxyuridine and in the sensitivity of all the thymidine phosphorylating activity to competition by BVdU. However, in contrast to the HSV TK, which phosphorylated deoxycytidine and iododeoxycytidine relatively efficiently and was sensitive to ACG, the HVS enzyme did not phosphorylate deoxycytidine or iododeoxycytidine and was insensitive to ACG. Whilst HVS, therefore, shares the characteristic of other members of the herpesvirus group of inducing a novel TK, the properties of the HVS-induced enzyme differ significantly from the enzyme of the prototype herpesvirus, HSV. The properties of the HVS TK are nonetheless sufficiently distinct from those of the uninfected cell to provide a possible basis for selective antiviral chemotherapy based on preferential phosphorylation of nucleoside analogues such as BVdU by infected cells. PMID- 6278058 TI - The surface antigens of orthopoxviruses detected by cross-neutralization tests on cross-absorbed antisera. AB - Cross-neutralization tests were done on accepted species and recently isolated members of the genus Orthopoxvirus using antisera which had been separately absorbed with the various viruses. The results provided evidence for the involvement of four neutralizing antigens, and their distribution among 13 virus strains was determined. Monkeypox (Congo-8-Lombe), camelpox (Gorgan), ectromelia (Mill Hill), 'Lenny' and elephant poxviruses had distinctive antigenic formulae. Lister and Wyeth vaccines were indistinguishable but different from Copenhagen and EM63 vaccines which were themselves distinct. Cowpox (Brighton), buffalopox (BP4), MK 10, and Moscow poxviruses were indistinguishable. Examples were found where viruses shared surface antigens but were not all neutralized by antibody to them. this reduced the practical value of the technique for virus identification. Evidence was also obtained for the existence in some viruses of a fifth antigen, antibody to which could block neutralization by antibody to one particular antigen. PMID- 6278059 TI - Inability to transmit scrapie by transfection of mouse embryo cells in vitro. AB - Infectivity of nucleic acid from highly infectious mouse scrapie brains was studied by transfecting the nucleic acid in vitro prior to inoculation into animals. Foetal mouse brain and whole mouse embryo cultures were chosen as the cells used for the transfection. As an internal control, infectious nucleic acid was recovered from bacteriophage phi X174 and whole virus particles were obtained when cultures were transfected with herpes simplex virus DNA. In contrast, none of the animals inoculated with nucleic acid preparations derived from scrapie infected tissues developed scrapie disease either with or without transfection procedures. These findings (i) show transfection techniques fail to elicit evidence of a scrapie-specific infectious nucleic acid and (ii) confirm the observation that scrapie is not a viroid. PMID- 6278060 TI - Gene dosage as a regulatory factor for gene expression. I. In lambda plac5 infected cells. AB - To study the effect of gene dosage on gene expression, lambda plac5cI857O29P3, a replication defective lambda phage carrying part of the lac operon (containing the lac promotor, operator and z gene) in the b2 region was studied in Escherichia coli strain JC6256 where the lac operon is deleted and at a temperature where the lambda repressor is inactive. In measuring the synthesis of beta-galactosidase, it was possible to separate the effects of the lac promoter from those of the phage promoter. When the synthesis of beta-galactosidase was initiated from the inserted lac promoter in JC6256(lambda +) in the presence of additional cyclic AMP, the rate and level of beta-galactosidase synthesis were directly proportional to the multiplicity of infection (gene dosage). Furthermore, beta-galactosidase synthesis was initiated about 5 min after infection, just as with isopropyl-beta-D-thiogalactoside (IPTG) induction. When the synthesis of beta-galactosidase was initiated from the phage promoter in JC6256 in the absence of additional cyclic AMP, the rate and level of beta galactosidase synthesis were again linearly proportional to gene dosage. On the other hand, initiation of beta-galactosidase synthesis was delayed until 10 to 20 min after infection. These results suggest that: (i) in the absence of negative controlling factors, the extent of gene expression is proportional to gene dosage; (ii) varying the gene dosage can be used to regulate gene expression. PMID- 6278061 TI - An IgM-producing B lymphoblastoid cell line established from lymphomas induced by a non-defective reticuloendotheliosis virus. AB - Chick syncytial virus (CSV), a strain of avian reticuloendotheliosis virus (REV) causes lymphoid tumours in chickens after a prolonged incubation period. A number of CSV-induced tumours were examined for cell surface antigen and were found to be of the B cell type and to produce immunoglobulin. Attempts were made to grow in vitro cell lines from CSV-induced tumours and a lymphoblastoid cell line was established from a liver tumour of a chicken that was inoculated with CSV via the yolk sac in embryo. The donor chicken was viraemic at the time the tumour was removed. The cell line is designated RECC-RP13, it produces non-defective REV, is a B cell type and it produces IgM. It is free from infection with endogenous and exogenous avian leukosis virus (ALV) and has an increased number of chromosomes. Sequences specific to REV were detected in at least four sites in cellular DNA from RECC-RP13. Sequences specific to ALV DNA, beyond that normally found in 151(5) X 7(1) cells, were not found in DNA from this cell line. PMID- 6278062 TI - Sulphated glycoproteins induced by herpes simplex virus. AB - BHK cells infected with strain 17 syn+ (HSV-1) or HG52 (HSC-2) incorporated inorganic sulphate into polypeptides which co-migrated on SDS-polyacrylamide gels with virus-induced glycoproteins. The major sulphated glycoprotein was glycoprotein E. In addition, less-intense sulphated bands co-migrated with glycoprotein D and HSV-1 glycoprotein A/B/C. Sulphate label co-migrating with HSV 2 glycoprotein A/B/C was occasionally observed. We have investigated which sulphated polypeptides are excreted from infected cells. Major ones of apparent mol. wt. 32000, 34000 and 35000 were excreted from cells infected with syn+. In addition, polypeptides which migrated in the vicinity of glycoprotein D were often excreted from cells infected with either 17 syn+ or HG52. The 32K, 34K and 35K polypeptides were antigenically related to glycoprotein D and over 95% of the total amount synthesized was excreted. Analysis of intracellular sulphated polypeptides using intertypic recombinants mapped glycoprotein E to between 0.832 and 0.950 units of the HSV genome. PMID- 6278063 TI - Mouse Elberfeld (ME) virus determines the cell surface alterations when mixedly infecting poliovirus-infected cells. AB - The surface alterations of HEp-2 cells induced by mixed infection with two different picornaviruses (poliovirus and ME virus) were compared by scanning electron microscopic and transmission electron microscopic studies and by 51Cr release assay. The contribution of each of the viruses to the resulting surface changes was discernible, as investigations on the chronology of the cytopathic alterations demonstrated that the changes were distinct for either virus. The surface of ME virus-infected cells was characterized by large membranous structures ('sheets' and blebs) representing huge vacuoles. These sheets were not seen in poliovirus-infected cells. Poliovirus induced more prominent cell pycnosis, elongation of filopodia and condensation of collapsed microvilli on the cell surface than ME virus. Mixed infection with these two viruses led to surface alterations typical for ME virus. These ME virus-specific changes occurred irrespective of poliovirus reproduction or its inhibition by guanidine. ME virus specific alterations also predominated in cytolytic membrane damage as expressed by 51Cr-release from infected cells. 51Cr-release was more pronounced from ME virus than from poliovirus-infected cells, even when ME virus reproduction was suppressed by interfering poliovirus. However, alteration of the internal structures of the infected cells was only dominated by ME virus when the reproduction of poliovirus was suppressed. PMID- 6278064 TI - The metabolic fate of independently initiated VSV mRNA transcripts. AB - The kinetics of synthesis and the metabolic stability of uncapped vesicular stomatitis virus (VSV) mRNA transcripts have been studied using techniques which clearly differentiate them from other RNA species. The triphosphate-initiating mRNA transcripts accumulate for at least 5 h during a typical in vitro transcription reaction. The great majority of these transcripts are smaller than a functional message and have been released from the template-transcriptase complex. Label that accumulates in them is stable and is not detectably diminished after a 1 h chase with unlabelled precursor. These kinetic properties are not these expected for active intermediates in mRNA synthesis and suggest that the uncapped transcripts are products of aborted transcription that accumulate during the transcriptive process. However, we cannot rule out that a small subset of these transcripts may be elongated into mature mRNA. Initiation of transcription at internal positions along the VSV genome is both frequent (one half to one-sixth as frequent as initiations at the leader RNA gene) and site specific (occurring only at the beginning of cistrons). The relevance of these results to the models for VSV transcription is discussed. PMID- 6278065 TI - Construction of a hybrid plasmid molecule containing the total coding region of Semliki Forest virus 26S mRNA. AB - We have constructed a hybrid plasmid molecule that contains the complete coding sequences from the 26S mRNA of Semliki Forest virus. Five fragments which together covered the mRNA sequence were isolated from three original hybrid plasmids and joined together at five different restriction endonuclease cleavage sites using T4 DNA ligase. PMID- 6278066 TI - Propagation of human hepatitis A virus in conventional cell lines. AB - Fecal extracts of hepatitis A (HA) patients were selected for the presence of hepatitis A virus (HAV) by radioimmunoassay (RIA) and immune electron microscopy (IEM). When FL and Vero cells were inoculated with fecal extracts containing HAV, development of hepatitis A antigen (HAAg) was evident in the cytoplasm of the two cell lines by the indirect immunofluorescence (IF) test. The antigen was detectable in the cells 12 hr postinoculation (pi), and reached a plateau within two days pi. FL cell cultures inoculated with a specimen containing HAV were harvested and passaged four times. During the passages, efficient production of HAAg was confirmed in the infected cultures by three different serological tests: The indirect IF test, RIA using fixed cells, and RIA by the sandwich method. At the second and fourth passages, HAV particles were recovered in abundance from infected FL cell cultures by IEM. Throughout these experiments, no cytopathic effect (CPE) was discernible in the cultures. PMID- 6278067 TI - The 30- to 54-nm rotavirus-like particles in gastroenteritis: incidence and antigenic relationship to rotavirus. AB - The 30- to 54-nm rotavirus-like particles were observed in the stool specimens of 17 children with gastroenteritis. These small rotavirus-like particles were shown to be antigenically related to the commonly described 68-nm rotavirus using the techniques of immune electron microscopy and ELISA (enzyme-linked immunosorbent assay). Four specimens containing the small rotavirus-like particles were shown to contain an antigen of a common human rotavirus serotype (type 2). The findings of small rotavirus-like particles of different diameters sharing a common antigen with rotavirus type 2 cautions against the naming of new candidate viruses based on morphology alone. In addition, the shedding of pure populations of single shelled rotaviruses, herein described, could be an unusual phenomenon which may occur only sporadically. The relationship of the smaller rotavirus-like particles to rotavirus morphogenesis is discussed. PMID- 6278068 TI - Effect of centrifugation on herpes simplex virus isolation. AB - The effects of high-speed centrifugation on the isolation of herpes simplex virus (HSV) were studied. Aliquots of laboratory or clinical specimens were inoculated into test tubes and flat-bottomed tubes containing HEp2 monolayers. Test tubes were incubated at 35 degrees C on roller drums (standard method), and flat bottomed tubes were centrifuged at 15,000g at 35 degrees C for 1 hr, before being incubated at 35 degrees C without rolling (centrifuged method). Centrifugation of clinical and laboratory specimens of HSV type 1 and HSV type 2 produced significantly increased isolation rates compared with the standard method. When clinical and laboratory specimens were diluted, the centrifuged method was more sensitive at all dilutions. When 20 specimens were used for end-point titrations, the centrifuged method was 10 times more sensitive for 15 specimens and 100 times more sensitive for five specimens. There was no difference in the time taken for the appearance of cytopathic effect (CPE) between the standard and centrifuged methods. PMID- 6278069 TI - Comparison of human cytomegalovirus growth in MRC-5 human fibroblasts, brain, and choroid plexus cells in vitro. AB - Cell cultures derived from human brain, choroid plexus, and human lung fibroblasts (MRC-5) were infected with the Towne strain of human cytomegalovirus (CMV). The cytopathic effect, beginning 24-48 hours after infection, was characterized by foci of enlarged rounded cells that spread slowly and eventually coalesced to destroy the entire monolayer within one week. Cowdry type A inclusion bodies and herpes virus nucleocapsids were seen in infected cells. CMV specific antigen was demonstrated by immunofluorescence in fibroblasts and astrocytic cells of brain cultures and in cells of choroid plexus cultures as well as in MRC-5 fibroblasts. Despite these morphologic and immunochemical similarities the growth of CMV differed in cells of brain and choroid plexus origin as compared with MRC-5 cells. In brain and choroid plexus cell cultures most of the virus remained cell-associated throughout the observation period of one week, whereas in MRC-5 cells the CMV was found in both cell-associated and cell-free fractions of harvested material. PMID- 6278070 TI - Further evidence for common antigens in herpes simplex and varicella-zoster viruses. AB - To elucidate the mechanism of heterologous antibody responses to herpes simplex virus (HSV) and varicella-zoster virus (VZV) which occur in some patients with HSV or VZV infections, stronger evidence was sought for the existence of cross reacting antibodies to these viruses, using antibody absorption procedures. Absorption of sera from initial HSV infections with HSV antigen was found to abolish heterologous antibody titer rises to VZV, as demonstrated in complement fixation, neutralization, and anti-complement immunofluorescence test systems. In most instances, convalescent-phase titers to heterologous VZV were reduced by HSV absorption to levels comparable to those in the acute-phase serum, indicating that cross-reacting antibodies were, in fact, responsible for the heterologous antibody titer rises. Absorption of convalescent-phase sera from HSV or VZV patients with homologous antigen also abolished or greatly diminished immunoprecipitating activity with the heterologous antigen, furnishing additional evidence of the existence of cross-reacting antibodies. Absorption of sera with insolubilized IgG to re-remove rheumatoid factor, which was present in a number of the sera studied, had no effect on either homologous or heterologous antibody titer increases. The demonstration of cross-reacting antibodies to HSV and VZV supports the concept that these two human herpesviruses share common antigen(s). PMID- 6278071 TI - Recovery of cytomegalovirus and Chlamydia trachomatis from vaginal tampons. AB - Herpes simplex virus (HSV), cytomegalovirus (CMV), and Chlamydia trachomatis are important agents in venereal and neonatal disease. Vaginal tampon culture for HSV has previously been demonstrated to be a simple and effective technique for quantitative culture of cervical secretions. We have evaluated the tampon culture as a means of performing quantitative cultures for CMV and C trachomatis. Cell free and cell-associated CMV were quantitatively recovered from vaginal tampons when extraction was performed within one hour of tampon inoculation. However, when tampons were stored, there was a rapid loss of infectivity over time at all storage temperatures except -70 degrees C. C trachomatis was quantitatively recovered from tampons stored at less than or equal to 4 degrees C for four days. When stored at -70 degrees C, C trachomatis was stable on tampons for more than one week. Because HSV, CMV, and C trachomatis are stable in a single transport medium, a tampon stored at 4 degrees C briefly or at -70 degrees C for one week could be utilized for the detection of all three agents. PMID- 6278072 TI - A two-part study of the aetiological role of rotavirus in intussusception. AB - The aetiology of intussusception is ill-defined, with viruses being incriminated as one of many possible aetiological agents. A two-part study was performed by us to investigate the aetiological role of rotavirus in intussusception. Retrospective epidemiological data revealed a negative correlation between the incidence of rotaviral gastroenteritis and the incidence of intussusception. A prospective investigation employing electronmicroscopy, enzyme-linked immunosorbent assay (ELISA), and immunofluorescence studies of faeces and fluorescent antibody studies of sera demonstrated evidence of rotavirus infection in only 2 of 24 children with intussusception. No evidence was forthcoming in this study of an aetiological role of rotavirus in intussusception. PMID- 6278073 TI - Restriction endonuclease analysis of varicella-zoster vaccine virus and wild-type DNAs. AB - The DNA from several clinical isolates of varicella-zoster virus (VZV) were compared with the DNA from the vaccine strain VZV using three restriction endonucleases: BamHI, BgII, and HpaI. When electrophoresed through an agarose gel, the vaccine DNA digestion pattern was significantly different from the digestion patterns of the wild-type DNAs. Variations in the digestion pattern of the separate clinical isolates were also observed. PMID- 6278074 TI - Enhanced responsiveness of alpha-noradrenergic receptors following discrete electrolytic lesions of the nucleus locus coeruleus: a behavioral demonstration. AB - Locomotor activity was evaluated in groups of reserpine-treated mice administered the catecholamine receptor agonists apomorphine and clonidine. Pretreatment of selected groups of mice with discrete electrolytic destruction of the nucleus locus coeruleus (LC) resulted in a clonidine-potentiation of apomorphine stimulated locomotor activity. This effect was greater in unilateral than bilateral lesioned mice. LC lesions also significantly reduced the level of locomotor habituation achieved within a 60 min observation period. The data are interpreted to suggest that discrete lesions of the nucleus LC results in increased alpha-noradrenergic receptor sensitivity. PMID- 6278075 TI - Testosterone decreases beta-adrenoceptor sites in rat pineal gland and brain. AB - Testosterone administration to orchidectomized rats brought about a significant, 55% decrease of beta-adrenoceptor sites in the pineal gland, assessed from the specific binding of radioactive dihydroalprenolol (DHA). The changes in density of binding sites were not accompanied by significant modifications of the Kd. FSH or LH treatment of acutely castrated animals did not affect pineal beta adrenoceptor binding. The depressive effects of testosterone in beta-adrenergic receptors were also observed in crude membrane fractions of medial basal hypothalamus and cerebral cortex. Sympathetic denervation of the pineal gland by superior cervical ganglionectomy did not abolish the changes in pineal beta adrenoceptor density caused by testosterone. Hormone effects did not depend on a direct effect of the hormone on beta-adrenoceptor sites because testosterone did not compete with [3H]-DHA for the binding sites, in vitro. These results suggest that testosterone depresses pineal beta-adrenergic sites by acting mainly on postsynaptic sites. PMID- 6278076 TI - Plasma beta-lipotropin levels in Parkinson's disease. AB - Previous reports have indicated that patients with Parkinson's disease have elevated plasma levels of immunoreactive (IR) beta-melanocyte stimulating hormone (beta-MSH), which may have implications as to its pathogenesis and treatment. Recent methodological advances, however, have demonstrated that what had originally been measured in human plasma as beta-MSH actually represents beta lipotropin (beta-LPH), and that beta-MSH as such does not normally circulate in human plasma. With the capacity to specifically measure immunoreactive beta-LPH in human plasma, we have determined plasma levels of immunoreactive beta-LPH as well as ACTH and prolactin in three groups of subjects: A. Parkinson patients untreated with levodopa (n = 11); B. Parkinson patients on levodopa therapy (n = 21); C. Controls (n = 6). No difference was found in plasma levels of IR-beta-LPH and IR-ACTH between these three groups. Plasma levels of prolactin were not different in either group of Parkinsonian patients as compared to controls. However, prolactin levels were significantly lower in the Parkinsonian patients treated with levodopa versus the untreated group. These data suggest that there is no defect in beta-LPH release from the pituitary in Parkinson's disease. PMID- 6278077 TI - The effect of ouabain on the release of [14C]acetylcholine and other substances from synaptosomes. PMID- 6278078 TI - Effect of homo-beta-proline and other heterocyclic GABA analogues on GABA uptake in neurons and astroglial cells and on GABA receptor binding. AB - Two groups of GABA (gamma-aminobutyric acid) analogues, one comprising derivatives of beta-proline and the other compounds structurally related to nipecotic acid, were investigated as potential inhibitors of high-affinity GABA transport in neurons and glial cells, as well as displacers of GABA receptor binding. In addition to cis-4-hydroxynipecotic acid, which is known as a potent inhibitor of GABA uptake, homo-beta-proline was the only compound which proved to be a potent inhibitor of glial as well as neuronal GABA uptake. IC50 values for GABA uptake into glial cells and brain cortex "prisms" were 20 and 75 micro M, respectively, and the IC50 value obtained for GABA uptake into cultured neurons was 10 micro M. A kinetic analysis of the action of homo-beta-proline on GABA uptake into cultured astrocytes and neurons showed that this compound acts as a competitive inhibitor of GABA uptake in both cell types. From the apparent Km values, Ki values for homo-beta-proline of 16 and 6 micro M could be calculated for glial and neuronal uptake, respectively. This mechanism of action strongly suggests that homo-beta-proline interacts with the GABA carriers. Furthermore, homo-beta-proline also displaced GABA from its receptor with an IC50 value of 0.3 micro M. The cis-4-hydroxynipecotic acid analogues, cis- and trans-4 mercaptonipecotic acid, had no inhibitory effect on glial or neuronal GABA uptake. Other SH reagents, PCMB, NEM and DTNB, were shown to be relatively weak inhibitors of GABA uptake into cultured astrocytes, suggesting that SH groups are not directly involved in the interaction between GABA and its transport carrier. PMID- 6278079 TI - Interactions of catecholestrogens with cytoplasmic and nuclear estrogen receptors in rat pituitary gland and hypothalamus. AB - The affinity of a series of catecholestrogens for 7S cytoplasmic receptor proteins from hypothalamus and pituitary gland of ovariectomised rats was assessed in vitro by a competitive charcoal binding assay at 4 degrees C. The equilibrium dissociation constants (Ki) of catecholestrogens 4-hydroxyestradiol, 4-hydroxyethynylestradiol, 2-hydroxyestradiol, 2-hydroxyethynylestradiol, and 4 hydroxyestrone were of the same order (Ki: 0.3-0.6 nM) as those of estradiol and ethnylestradiol (Ki: 0.1 nM). Methylation of 2-hydroxyestradiol led to a substantial loss of binding affinity. Tritium-labelled receptor complexes were demonstrated in KCl extracts of purified nuclei from pituitary and hypothalamic tissue 1 h after intravenous injection of 0.1 mCi tritiated 2- or 4 hydroxyestradiol. These macromolecular complexes sedimented in the 5-6S region of 5-20% (w/v) sucrose gradients containing 0.4 M-KCl. Further evidence for the translocation of estrogen receptors by catecholestrogens into the nuclei of rat pituitary and hypothalamus was the increase in nuclear receptor concentrations, measured by exchange assay, 1 h after the intraperitoneal injection of 0.1 mg unlabelled catecholestrogen. Administration of 4-hydroxyestradiol and 4 hydroxyethynylestradiol increased nuclear receptor concentrations to the same maximal levels as those following application of the same dose of estradiol or ethynylestradiol, whereas the respective 2-hydroxylated compounds exhibited only 60-70% of the maximal translocating capacity. The in vivo translocating capacities of the various catecholestrogens tested at this dose correlated well with their binding affinities for cytosol receptors determined in vitro. PMID- 6278080 TI - Dermorphins, opioid peptides from amphibian skin, act on opioid receptors of mouse neuroblastoma x rat glioma hybrid cells. AB - Dermorphin and its Hyp6 analogue are opiate-like heptapeptides originally discovered in frog skin and characterized by the presence of a D-Ala2 residue in their sequence. They were assayed for their capacity to compete with [3H]Leu enkephalin for binding to opioid receptors in membranes of neuroblastoma x glioma hybrid cells. In the presence of 7 nM-[3H]Leu-enkephalin, the concentrations at which they caused 50% inhibition of [3H]enkephalin binding (IC50 values) are 0.1 micro M and 0.3 micro M, respectively. In contrast, the synthetic L-Ala2 dermorphin shows very low affinity for the opioid receptors. In addition, like other opioid peptides, dermorphin and hyp6-dermorphin inhibit the elevation by prostaglandin E1 (PGE1) of the level of adenosine 3':5'-cyclic monophosphate (cyclic AMP) (IC50 values 0.2 micro M and 0.4 micro M, respectively). The inhibition is prevented by the opiate antagonist naloxone, L-Ala2-dermorphin is at least three orders of magnitude less potent in inhibiting the PGE1-evoked increase in the level of cyclic AMP. The results show that peptides with an amino acid sequence quite different from that of the enkephalins can bind to opioid receptors of the hybrid cells. PMID- 6278081 TI - Cerebral synaptic transmission during anoxia is protected by creatine. AB - Synaptic transmission in cerebral tissue fails very rapidly in the absence of oxygen; the metabolic basis for this is not known. We report here that the transmission failure in the guinea pig hippocampal slice can be delayed threefold by exposing the tissue to extracellular creatine (Cr) for 3 h. The improved survival is associated with an increase of tissue phosphocreatine (PCr) concentration. These data argue that the metabolic basis for synaptic transmission failure is a fall in tissue ATP concentrations. They also indicate a way to protect brain tissue against anoxic damage. PMID- 6278082 TI - The effect of visual deprivation on beta-adrenergic receptors in the visual centres of the rat brain. AB - The levels of binding of [3H]dihydroalprenolol to beta-adrenergic receptors in the visual centres and frontal cortex from brains of control, dark-reared and monocularly deprived rats were compared. Receptor binding is changed in monocularly deprived rats in the lateral geniculate nuclei and superior colliculi of both sides. Scatchard analyses indicated that the changes in the [3H]dihydroalprenolol binding in the lateral geniculate nuclei were due to alterations in both receptor affinity and receptor number. No effect of dark rearing could be detected. PMID- 6278083 TI - The binding of the GABA agonist [3H]THIP to rat brain synaptic membranes. AB - THIP (4,5,6,7-tetrahydroisoxazolo[5,4-c]pyridin-3-ol) is a specific GABA agonist with potent analgesic properties. The binding of radioactive THIP to thoroughly washed, frozen, and thawed membranes isolated from rat brains has been studied at 2 degrees C under sodium ion-free conditions and compared with the binding of [3H]GABA and [3H]piperidine-4-sulphonic acid ([3H]P4S). The best computer fits to the experimental data were in all cases attained with a receptor model based on three independent binding sites, of which only the high- and medium-affinity sites could be characterised satisfactorily. While the KD values were found to be comparable for all three ligands employed, the density of the high-affinity binding site (BM1) was, with the exception of the membranes from the cerebellum, considerably lower for [3H]THIP than for [3H]GABA and [3H]P4S. The regional distribution of the GABA receptors, which bind [3H]THIP, was different from those recognizing [3H]GABA and [3H]P4S. A number of analogues, including asymmetric compounds with known configuration, were tested as inhibitors of the binding of [3H]GABA, [3H]muscimol, [3H]THIP, [3H]isoguvacine, and [3H]P4S. The concentrations of the asymmetric compounds required for the inhibition of [3H]P4S binding were much higher than those required for the displacement of [3H]GABA, [3H]muscimol, [3H]THIP, and [3H]isoguvacine. The comparable relative potencies of inhibitors do, however, indicate that all of the ligands bind to the GABA receptors. PMID- 6278084 TI - Opiate receptor-mediated inhibition of adenylate cyclase in rat striatal plasma membranes. AB - Plasma membranes from rat striatum contain adenylate cyclase activity that is subject to dual regulation by GTP. Low concentrations (up to 30 nM) of the nucleotide increase activity whereas higher concentrations evoke a steady decline in activity; such behavior characterizes dually regulated adenylate cyclase systems. The opiates, morphine sulfate and D-Ala-Met-enkephalin, produce naloxone reversible inhibition of the enzyme that is dependent on "inhibitory concentrations" of GTP (above 50 nM). In the absence of GTP no inhibition is observed. Sodium ions decrease the inhibition of activity promoted by GTP alone, but amplify the degree of inhibition seen in the presence of the opiates and GTP. The potencies of the opiates in mediating these effects mirror their affinities for delta opiate receptors in striatum. It is suggested that this action of the opiates may represent their primary action in striatum. PMID- 6278086 TI - ACTH1--24 releases a protein from synaptosomal plasma membranes. AB - Brain membranes contain several protein kinases, all of which appear to play a role in the regulation of neuronal functioning. These membranes also contain numerous (phospho) proteins. It has been proposed that the degree of phosphorylation of some of these proteins may affect neuronal membrane properties. In a series of previous reports we showed that ACTH1--24 inhibits the endogenous phosphorylation of several synaptosomal plasma membrane (SPM) proteins including the B-50 protein. Although we have speculated that the degree of phosphorylation of B-50 may be important in regulating the turnover of membrane (poly)-phosphoinositides, the exact nature of the interaction between ACTH1--24 and B-50/B-50 protein kinase is unknown. The purpose of the present study was to determine whether treatment of SPM with ACTH1--24 will lead to a specific release of proteins from SPM. We found that ACTH1--24 specifically releases a 41,000 Mr protein from rat brain SPM. Although we are not certain about the biological significance of the release of this polypeptide, it is of sufficient interest for further research in view of the lack of success of finding binding of labeled ACTh to brain membranes. PMID- 6278085 TI - Molecular interactions of etazolate with benzodiazepine and picrotoxinin binding sites. AB - Pyrazolopyridines, such as etazolate (SQ 20009), enhance [3H]diazepam binding to a Lubrol-solubilized fraction that has specific binding sites for 3H benzodiazepines, [alpha-3H]dihydropicrotoxinin (DHP) and [3H]muscimol. Etazolate enhancement of [3H]diazepam binding was inhibited by picrotoxinin. Furthermore, etazolate inhibited the [3H]DHP binding in a Lubrol-solubilized fraction with an IC50 value of 6-8 microM. These results provide evidence that etazolate, like pentobarbital, modulates benzodiazepine binding via the DHP-sensitive site of the benzodiazepine-GABA receptor-ionophore complex. PMID- 6278087 TI - Poly(adenylate) polymerase activity of rat brain. AB - The poly(adenylate)[poly(A)] polymerase of rat brain, as in rat liver, is located primarily in the nuclear sap when nuclei are prepared under hypertonic conditions. The enzyme can be released from nuclei in two forms. Form I is prepared by gentle incubation of nuclei at 0 degrees C in hypotonic buffer. It has a Mn optimum of 0.6 mM and a pH optimum between 8 and 9. The ATP concentration curve plateaus at 0.2 mM. The optimal poly(A) primer concentration is 600 micrograms/ml, which is three times higher than that for the enzyme similarly prepared from liver. The time course of the reaction for the form I enzyme is increasing over the first 40 min and becomes nearly linear thereafter. Form I is not stimulated by either calcium or cyclic nucleotides, but is inhibited by polyamines, pyrophosphate, and high concentrations of GTP. Form II enzyme is prepared by homogenization of nuclei in hypotonic buffer. It has the same ATP and poly(A) optima as the form I enzyme but displays linear kinetics over a 60-min time course. It is slightly stimulated by cGMP and cAMP and strongly inhibited by spermine, sodium pyrophosphate, and high concentrations of GTP. PMID- 6278088 TI - Melittin stimulates incorporation and degradation of sphingomyelin in synaptosomal plasma membranes. AB - Melittin enhanced sphingomyelin (SPM) degradation by the neutral membrane-bound sphingomyelinase from calf brain synaptosomal plasma membranes (SYM) up to 20 fold. Melittin in concentrations as high as 100 microM did not significantly alter membrane fluidity of SYM as measured by fluorescence depolarization and electron spin resonance (ESR) using diphenylhexatriene and a doxyl derivative of SPM, respectively. In the concentration range 100--1000 microM, melittin was observed to rigidify SYM. The incorporation of SPM derivatives into the lipid bilayer of SYM was demonstrated by ESR measurements. Melittin enhanced the uptake of SMP-derivatives into SYM. PMID- 6278089 TI - The distribution of radioligand binding activity and 5'-nucleotidase activity in bovine caudate nucleus subcellular fractions. AB - The distribution of 5'-nucleotidase activity, dopaminergic [3H]spiperone binding sites, and [3H]quinuclidinyl benzilate (QNB) binding sites in different subcellular fractions of bovine caudate nucleus has been studied. Each activity was enriched in a microsomal (P3) preparation from that tissue. The microsomal preparation was further fractionated by different techniques. First, the P3 fraction, or a sonicated P3 fraction, was fractionated on a discontinuous sucrose density gradient. Second, the P3 fraction, or a digitonin pretreated P3 fraction, was fractionated on a continuous sucrose density gradient. The results obtained demonstrate that 5'-nucleotidase activity does not cofractionate with radioligand binding activity, although no difference between the distributions of [3H]spiperone binding and [3H]QNB binding were seen. It is concluded that the two radioligand binding activities are located on nonglial membranes. PMID- 6278090 TI - Neurotransmitter receptor ligand binding and enzyme regional distribution in the pigeon visual system. AB - The relative importance of acetylcholine, dopamine, endogenous opiates, gamma aminobutyric acid (GABA), glutamate, glycine, noradrenaline, and serotonin as transmitters in the pigeon visual system was estimated by measuring the activity of choline acetyltransferase (ChAT), glutamic acid decarboxylase (GAD), and aromatic amino acid decarboxylase (AAD) as well as the binding of dihydroalprenolol, etorphine, kainic acid, muscimol, serotonin, spiroperidol, strychnine, and quinuclidinyl benzilate (QNB) in the tectum opticum, nucleus rotundus, ectostriatum, dorsolateral thalamus, and hyperstriatum (Wulst). As a nonvisual reference structure, the paleostriatal complex was included in the examination. The regional distribution of most of these parameters was very similar to data reported in the mammalian CNS supporting the hypothesis that the avian tectofugal and thalamofugal visual systems are homologous to the mammalian tecto-thalamo-cortical and retino-geniculo-striate pathways, respectively. On the basis of the low values of their parameters, some transmitters can be excluded as significant contributors in a number of structures. As a hypothesis for further investigations, the presence of cholinergic and serotoninergic systems in the Wulst, possibly originating in the dorsolateral thalamus and nucleus raphe, respectively, and of glycinergic and dopaminergic terminals in the paleostriatal complex is proposed. PMID- 6278091 TI - Early stimulation of phosphatidylcholine biosynthesis during Wallerian degeneration of rat sciatic nerve. AB - Phospholipid metabolism was studied in rat sciatic nerve during Wallerian degeneration induced by crush injury. Portions of crushed sciatic nerve, incubated with labeled substrates, showed significantly higher phosphatidylcholine synthesis than normal nerve, prior to any measurable alterations of phospholipid composition. Maximum synthesis occurred 3 days after crush injury, at which time the metabolism of other phospholipids was unchanged. After a rapid decrease in biosynthetic activity, a second phase of enhanced phosphatidylcholine synthesis occurred, beginning 6 days after crush injury. Increased incorporation of [33P]phosphate, [2-3H]glycerol, and [Me-14C]choline indicated stimulation of de novo synthesis of phosphatidylcholine 3 days after injury. Neither base exchange reactions nor sequential methylation of ethanolamine phospholipids contributed significantly to phosphatidylcholine synthesis. Assay of certain key enzymes under optimal conditions in subcellular fractions of sciatic nerve revealed higher activities of cholinephosphate cytidyltransferase, choline phosphotransferase, and acyl CoA:lysophosphatidylcholine acyltransferase in injured nerve, while choline kinase activity remained unchanged. This indicates that stimulation of phosphatidylcholine synthesis occurs via the cytidine nucleotide pathway, as well as by increased acylation of lysophosphatidylcholine. Although the cause of stimulated phosphatidylcholine synthesis remains unexplained, it is possible that trace amounts of lysophospholipids or other metabolites produced by injury enhanced phospholipase activity may be responsible. PMID- 6278092 TI - Highly activatable adenylate cyclase in [2-3H]adenine-prelabeled vesicles prepared from guinea pig cerebral cortex by a simplified procedure. AB - An improved procedure utilizing a simple fractionation technique is described for the preparation and use of [2-3H]adenine-prelabeled vesicles from guinea pig cerebral cortex containing highly responsive adenylate cyclase activity. Adenosine consistently increased activity 1500--2000%, contrasted with activations of 200--300% previously reported by other investigators. Adenosine at 5 microM was more active in our system than at 20 times this concentration in studies by other investigators, increasing activity by 580--840%. Experimental conditions were explored, and Ca2+ was found to be necessary during tissue homogenization, but not during subsequent vesicle incubation. However, neither the higher Ca2+ concentration used by us (2.5 mM) nor the method of tissue homogenization could adequately explain the high activity of our preparations. The size of the incubation vessel was critical for both low basal activity and high activity in the presence of adenosine. Our preparations were similar to others in that combinations of neurohormones, which included histamine and epinephrine, elicited higher activities than individual neurohormones. Inspection of our vesicle preparations by scanning electron microscopy also showed them to be compatible with previously described preparations. PMID- 6278093 TI - Angiotensin converting enzyme in Alzheimer's disease increased activity in caudate nucleus and cortical areas. AB - The activity of the dipeptidyl carboxypeptidase, angiotensin converting enzyme, was assayed in several brain regions of patients dying with Alzheimer's disease and compared to that of appropriately age-matched controls. Enzyme activity was found to be elevated by 44% and 41% in the medial hippocampus and parahippocampal gyrus, respectively, and by 27% and 29% in the frontal cortex (area 10 of Brodman) and caudate nucleus, respectively, in Alzheimer's disease patients. Converting enzyme activity did not differ from controls in the nucleus accumbens, substantia nigra, temporal cortex, anterior or posterior hippocampus, amydgala, and septal nuclei. PMID- 6278094 TI - Effect of contraction level and magnitude of stretch on tonic stretch reflex transmission characteristics. AB - Electromyogram tonic stretch reflex responses were recorded from biceps brachii muscles in normal and cerebral palsied subjects sustaining either 10% or 20% of maximum voluntary contraction and attempting to keep the elbow stiff in a fixed position. The muscle was stretch by a sinusoidal perturbation applied by the experimenter to the elbow angle. Five different amplitudes of stretch were employed ranging 1.67 to 10.0 degrees peak to peak variation of elbow angle. Spectral analysis of the rectified and filtered electromyogram revealed "noisy" sinusoidal reflex responses with negligible harmonic distortion but the amplitude of the reflex responses did not increase linearly with the amplitude of stretch. An analysis of variance showed that for both groups of subjects the gain of the tonic stretch reflex increased significantly (p less than 0.001) with contraction level and decreased significantly (p less than 0.001) with magnitude of stretch. This finding illustrates that both magnitude of stretch and level of contraction need to be carefully controlled when measures of tonic stretch reflex responses are used to assess changes of muscle tone. PMID- 6278095 TI - Hepatocellular carcinoma with cranial metastasis and hyperglobulinaemia. PMID- 6278096 TI - Muscular inhibitory and excitatory phenomena during spike-and-wave discharges: effect of posture. PMID- 6278097 TI - Angiofibrosis of the brainstem in a woman with motor neuron disease. PMID- 6278098 TI - Subacute sensory neuropathy with remission: an association with lymphoma. AB - A case of subacute sensory neuropathy associated with Hodgkin's disease improved as the tumour was treated. The poor prognosis usually associated with this neuropathy may be attributable to its more common association with untreatable tumours. PMID- 6278099 TI - Buckthorn neuropathy in vitro : evidence for a primary neuronal effect. AB - The neuropathy produced by toxins isolated from the buckthorn shrub (Karwinskia humboldtiana) has previously been proposed as a model for primary PNS demyelination in vivo. These toxins have been suggested to act through a metabolic effect on the Schwann cell. The aim of the present study was to use myelinated organotypic cultures of nervous tissue to dissect further the pathologic effects of these compounds. Combination cultures of mouse spinal cord and dorsal root ganglia were exposed to buckthorn toxins T/496 or T/544 at doses of 1-5 microgram/ml for periods of 48 hours (h) to 14 days. Ultrastructural findings from toxin-treated cultures indicated a primary effect upon the PNS axon, both myelinated and unmyelinated. The effect was expressed mainly as a widening of the periaxonal space, and also as a redistribution of most axonal organelles to a marginal position within the axon or to their close association with stacks of smooth endoplasmic reticulum, leaving more central regions occupied largely by neurofilaments. These changes were followed by focal axonal swelling, floccular disruption of the axoplasm and, ultimately, Wallerian degeneration. Myelin degeneration and chromatolytic changes in ganglion cell bodies appeared to be secondary to axonal alterations. This tissue culture model reproduces many of the features of buckthorn intoxication in vivo. However, the present results suggest that the toxic effect is primarily upon the axon rather than the Schwann cell, and would appear to be consistent with an abnormality in axoplasmic transport. PMID- 6278100 TI - Cytochemical heterogeneity of the glial plasma membrane: 5'-nucleotidase in retinal Muller cells. AB - Localization of 5'-nucleotidase in the retina of four rodent species was investigated using histochemical and cytochemical techniques. It was revealed by light microscopy that the enzyme is present in the receptor layer, the external nuclear layer and the external plexiform layer. It was found by electro microscopy that the glial cells of the Muller type have a strong activity on their plasmalemma in the enzyme-positive layers. With the exception of weak and scattered activity on some Muller cell processes forming horizontal fibres and the honeycomb meshwork in the ganglion cell layer, none of the internal layers contain 5'-nucleotidase. The asymmetric distribution of this ectoenzyme on the surface of the glial cells is a novel finding and demonstrates a surprising heterogeneity of glial plasmalemma. PMID- 6278101 TI - Lateral spread of neural excitation during microstimulation in intermediate gray layer of cat's superior colliculus. AB - 1. Synaptically mediated spread of excitation has been studied during microstimulation in the intermediate gray layer of the superior colliculus of cats anesthetized with ketamine. Antidromic activation was used to identify those neurons sending axons to or through the contralateral pontine reticular formation. 2. Current thresholds were dependent on pulse duration, train length, and distance from the cell to the stimulus site. Stimulation with four cathodal pulses, 0.5 ms in duration, 30 microA in intensity, delivered at 400 Hz, excited cells of the intermediated gray up to 2-3 mm from the stimulus site. The results suggest that at least half the output cells in a region about 3 mm in diameter were driven by this stimulus. 3. The extent of spread in the unanesthetized midpontine-pretrigeminal cat was as great as, or greater than, that in animals anesthetized with ketamine. 4. Quick eye movements were evoked in ketamine anesthetized cats by 100-ms trains of 0.5-ms pulses delivered at 400 Hz. Current thresholds for eye movements ranged from 15 to 90 microA, with most falling below 25 microA. 5. These results suggest that intracollicular microstimulation, with stimuli commonly used in studies of electrically evoked saccades, is accompanied by widespread synaptic activation of the intermediate gray layer. Since the metrics of electrically evoked saccades seem, nonetheless, to depend primarily on the location of the stimulating electrode, information about amplitude and direction must somehow be encoded in the distribution of neuronal discharge. 6. One possible form of such a distributed coding mechanism is discussed. This model assumes that the spatial densities of cells projecting to vertical and horizontal pulse generators of the saccadic system vary systematically beneath the retinotopic collicular map. Signals to the pulse generators change in magnitude as the collicular discharge zone occupies different positions in the connectional gradients and engages the specific output systems in varying proportion. PMID- 6278102 TI - [14C]2-deoxyglucose uptake in ground squirrel brain during hibernation. AB - Autoradiographic patterns of [14C]2-deoxyglucose uptake are described throughout the brains of hibernating and euthermic ground squirrels. Autoradiographs of the brains of hibernating animals are generally homogeneous in comparison to euthermic animals; hence, the relative 2-deoxyglucose uptake (R2DGU) of gray to white matter for the majority of the 85 neural structures examined decreases during hibernation. Two categories of structures are identified as potentially important in hibernation: (1) structures that have the highest R2DGU during hibernation (cochlear nucleus, paratrigeminal nucleus, and superior colliculus) and (2) structures that undergo the least reduction in R2DGU in the transition from euthermia to hibernation (suprachiasmatic nucleus and lateral septal nucleus). The percentage of reduction in R2DGU that a structure undergoes in the transition from euthermia to hibernation is proportional to the R2DGU of that structure during euthermia. The suprachiasmatic, paratrigeminal, and cochlear nuclei undergo less of a reduction than would be predicted from this relationship and may be particularly important during hibernation. Sensory nuclei that receive primary afferent projections are among the structures with the highest R2DGU during hibernation. These metabolically active structures may be responsible for the sensitivity of the hibernator to environmental stimuli. PMID- 6278103 TI - Protein phosphorylation during afterdischarge in peptidergic neurons of Aplysia. PMID- 6278104 TI - Neurotrophic regulation of two properties of skeletal muscle by impulse-dependent and spontaneous acetylcholine transmission. PMID- 6278105 TI - Energy-requiring cell functions in the ischemic brain. Their critical supply and possible inhibition in protective therapy. PMID- 6278106 TI - Herniated nerve root as a complication of spinal tap. Case report. PMID- 6278107 TI - Secondary adrenal insufficiency after intrathecal steroid administration. AB - A 38-year-old man developed secondary adrenal insufficiency as a consequence of intrathecal methylprednisolone administration. Evidence in support of this diagnosis included an absent plasma cortisol response to insulin-induced hypoglycemia, an inadequate adrenal response to exogenous corticotropin stimulation, a typical delayed response to prolonged corticotropin infusion over 3 days, and the findings of an elevated level of prednisolone in the cerebrospinal fluid a full 2 months after its administration. It is therefore recommended that patients receiving intrathecal steroids be carefully observed for the possible development of secondary adrenal insufficiency. PMID- 6278108 TI - Femoral neuropathy caused by an iliopsoas bursal cyst. Case report. PMID- 6278109 TI - Malignant cerebellar gliomas. Report of 4 cases with special reference to tissue culture study. AB - The rare malignant cerebellar gliomas have been considered as an unusual form of brain glioblastomas. However, different forms have been described which can be divided into three groups: (a) cerebellar astrocytomas with malignant features; (b) undifferentiated tumors mimicking the cerebral glioblastomas; (c) malignant glioblastomatous recurrences of primary benign cerebellar astrocytomas. The present series is composed of four cases studied with silver impregnation or with tissue culture and time-lapse cinematography. It appears from this study that the cerebellar glioblastomas bear no relationship to the cerebral glioblastoma. Silver impregnation shows many astrocytes taking part in the tumor cell population and the behavior in vitro is similar to that of the common benign astrocytomas from which they seem to be derived. Thus, a sequence can be established: benign cerebellar astrocytoma, malignant cerebellar astrocytoma, cerebellar glioblastoma. PMID- 6278110 TI - Tc-99m glucoheptonate in detection of lung tumors. AB - Sixty-three patients aged 21-80 yr were examined. Tc-99m glucoheptonate (Tc-GH) scintigraphy was performed in 32 patients: 26 with primary lung carcinoma; six with metastases in lung, mediastinum, and pleura from carcinomas elsewhere; eight with benign pulmonary diseases; and 23 without known pulmonary disease. Tc-GH accumulated in 23 of 26 primary pulmonary carcinomas as active foci. The specificity of Tc-GH scans for neoplasm detection was higher than that of chest radiographs. The visualization of malignant tumors was much better in the late Tc GH images (5-6 hr) than in the early (1 hr). Metastases from other carcinomas were positive in four of six patients, but they were considerably better detected in the radiographs, except in one patient with metastatic hepatocellular carcinoma. Neoplasms or their metastases in the hilar and mediastinal regions were better detected in the Tc-GH scans than in the chest radiographs. Only one of eight benign lung processes was visualized (as a weak diffuse accumulation of Tc-GH in hilar scar formation), and 23 patients without pulmonary disease had no pathological foci. PMID- 6278111 TI - Mechanism of inhibitory effect of unavailable carbohydrate on intestinal calcium absorption. AB - The effect of unavailable carbohydrates on the intestinal absorption of calcium was studied in rats raised for 7 or 8 weeks on diets containing 10 or 20% of cellulose, glucomannan, or pullulan. The following results were obtained a) Body weight gain was diminished more severely in glucomannan groups than in cellulose groups. b) Serum calcium levels were slightly lower in all groups fed unavailable carbohydrates, whereas serum inorganic phosphorus levels were similar to that of the control group. c) There was a significant reduction of bone ash from rats fed glucommanan or cellulose with 620 glycoside bonds. d) Calcium transport measured in the everted duodenal sac remarkable decreased in the glucomannan 20% group, but slightly increased in the cellulose groups. e) Calcium binding activity was lowered significantly in all groups fed unavailable carbohydrates. f) Alkaline phosphatase and sucrase activities in the duodenum were also markedly decreased by prolonged intake of unavailable carbohydrates. These results indicate that inhibitory effect of unavailable carbohydrates on intestinal calcium absorption is partially due to the loss of calcium binding protein caused by gastrointestinal transit of large amounts of undigested substances. PMID- 6278112 TI - Disorders of cholecalciferol metabolism in old egg-laying hens. AB - It has been reported that the rate of cracked or soft-shelled eggs markedly increases in old laying hens. We investigated the effect of age on cholecalciferol metabolism in different age groups of laying hens. The egg production rate in hens more than 500 days old was maintained within a range of about 70% of that in young hens (230-320 days old), whereas the rate of cracked or soft-shelled eggs increased markedly with age. When kidney homogenates from the different age groups were incubated with [3H]-25-hydroxyvitamin D-3, renal 25 hydroxyvitamin D-3-1 alpha-hydroxylase activity was found to decrease markedly with age. When birds were given intravenously either [3H]-25-hydroxyvitamin D-3 or [3H]-1 alpha,25-dihydroxyvitamin D-3, the accumulation of [3H]-1 alpha,25 dihydroxyvitamin D-3 in plasma and target tissue also decreased with age. Forced molting performed in old hens restored eggshell quality. The treatment also restored, though partially, the in vivo accumulation of [3H]-l alpha,25 dihydroxyvitamin D-3 in the target tissues. These results suggest that the increased rate of cracked or soft-shelled eggs seen in older birds is associated with disorders of vitamin D-3 metabolism. PMID- 6278113 TI - Dietary Pectin's effect on starch utilization in rats. AB - Dietary pectin's effects on intestinal 14C-starch absorption, oxidation to 14CO2 and intestinal weight were studied. Rats were adapted to either a 5% pectin diet or a control diet for 10 or more days before test meals of the same composition but containing 14C-starch were fed., One hour after the test meal, more 14C label remained in the stomach as well as the entire gastrointestinal tract of pectin fed rats. Also, less 14C had reached the distal quarter of the small intestine at one hour. Rats eating the 5% pectin diet had higher wet and dry weights of the small intestine. A separate group of pectin-fed rats exhaled more 14CO2 than controls during the first hour after the test meal. Amount of 14C in the gastrointestinal tract and exhalation of 14CO2 did not differ between diet groups at 2, 3, or 4 hours after the test meal. Results indicate that dietary pectin caused a decrease in starch absorption during the fist hour after the meal and it led to an increase in small intestinal weight. Dietary pectin also resulted in a higher percentage of ingested 14C appearing as 14CO2 during the first hour after the test meal. PMID- 6278114 TI - Dietary wheat bran lowers colonic pH in rats. AB - The effect of chronic dietary wheat bran supplementation on the pH of gastrointestinal contents was examined in thirty-one Sprague-Dawley rats. Ten control rats received a fiber-free diet while twenty-one rats were fed the same diet plus a 20% wheat bran supplement. The fiber-fed rats developed a greater acidity of intestinal contents: cecal pH was 6.30 +/- 0.33 in the controls (P less than 0.01). These changes in gastrointestinal pH may be of relevance to the nutrient modulation of gastric and colonic tumor development. PMID- 6278115 TI - Simplified assay of vitamin D2 in fortified dried milk by using two steps of high performance liquid chromatography. AB - A simplified method for the determination of vitamin D2 in fortified dried milk was established by using two steps of high-performance liquid chromatography (HPLC). About 1 g of fortified dried milk was accurately weighed and directly saponified. The extracted unsaponifiable matter was first subjected to preparative HPLC using a Nucleosil 5C18 column (reversed-phase type) with acetonitrile-methanol (1:1) as a mobile phase and a fraction containing vitamin D2 was separated. The separated fraction was subsequently subjected to analytical HPLC using a Zorbax SIL column (straight-phase type) with 0.4% isopropanol in n hexane as a mobile phase. Since the peak corresponding to vitamin D2 was clearly observed with separation from other concomitants on the chromatogram of the analytical HPLC, the vitamin was assayed by estimating the peak height. The overall recovery of vitamin D2 by the proposed method was 94.3 +/- 2.3% (mean +/- SD). Naturally occurring vitamin D3 derived from cow's milk was negligible in commercial fortified dried milk sold in Japan. When the proposed method was applied to 5 kinds of commercial fortified dried milk, satisfactory results were obtained. PMID- 6278116 TI - Tannic acid-glutaraldehyde fixative and pleomorphic adenomas of the salivary gland: an ultrastructural study. AB - Four cases of pleomorphic adenomas of parotid glands were fixed by immersion in a mixture of 4% tannic acid and 3% glutaraldehyde followed by OsO4 postfixation and heavy metals staining. This clearly visualized the presence of dense amorphous elastic fibers in cellular and myxochondroid areas. The dense amorphous fibers were associated with varying numbers of peripheral filaments which appeared sometimes as bundles of parallel fibrils without intermingled amorphous material. These intermediate-sized filaments were seen indirect continuity with similar intracellular filaments with or without interposition of basal-membrane-like material and/or with extracellular filament-containing vesicles. It is suggested that these filaments are synthesized intracellularly and released into the extracellular space by neoplastic myoepithelial cells mainly by cytoplasmic vesicle separation. In addition, small round vesicles with a dense core of amorphous material and staining characteristics identical to those of extracellular elastin were frequently seen in the peripheral parts of these myoepithelial-like cells. It is possible that these dense-cored vesicles have a secretory function and contain a precursor of elastin. PMID- 6278117 TI - Severe postural hypotension in childhood with autonomic neuropathy and occult systemic neuropathy. PMID- 6278118 TI - Treatment of severe neonatal infections with cefotaxime. Efficacy and pharmacokinetics. AB - We studied the pharmacokinetics and efficacy of cefotaxime in 32 neonates with severe gram-negative infections. Many of these patients had been treated unsuccessfully with combinations of antibiotics. Eighty-one percent of these patients were cured, 6% improved, and 13% had treatment failures; there were three deaths. Eighteen patients received cefotaxime alone; 16 were cured and two improved. These data indicate an efficacy of cefotaxime sufficient to warrant more rigorous future trials. The elimination half-life of cefotaxime ranged from 2.0 +/- 0.4 hours in term neonates more than one week of age to 5.7 +/- 0.8 hours in preterm neonates less than one week of age. A volume of distribution of approximately 0.63 L was similar for all infants irrespective of age and maturity. These kinetic data can be used in design of future therapeutic regimens in more rigidly controlled trials assessing indications for cefotaxime therapy in neonates. We recommend dosing as follows, using a dose of 25 mg/kg: every 12 hours for preterm infants less than one week of age, every 8 hours for preterm infants one to four and term infants less than one week of age, and every 6 hours for term infants more than one week of age. PMID- 6278120 TI - Synthesis of collagenolytic enzymes and their inhibitors by gingival tissue in vitro. Effect of endotoxin. PMID- 6278119 TI - Wilms tumor with aniridia/iris dysplasia and apparently normal chromosomes. AB - Two patients with Wilms tumor, iris dysplasia (complete aniridia in one and subtle iris defects in the other), normal karyotypes, and no gene loss demonstrable by enzyme marker and direct DNA analyses are presented. The findings indicate that aniridia and less severe iris defects define a risk for Wilms tumor even in the absence of del (11p13), and that there is as yet no consistent biochemical genetic marker for the aniridia-Wilms tumor association. PMID- 6278121 TI - Pituitary-adrenocortical stimulation and anti-inflammatory actions of 4-ethoxy-2 methyl-5-morpholino-3(2H)-pyridazinone (M73101). AB - We investigated the mechanism of the elevation of serum corticosterone level by 4 ethoxy-2-methyl-5-morpholino-3(2H)-pyridazinone (M73101) and its participation in the anti-inflammatory action. M73101 when given to rats intraperitoneally at doses of 50-200 mg/kg, caused an increase in serum and adrenal corticosterone in a dose-dependent manner. Oral administration of 200 mg/kg of M73101 also elevated the serum corticosterone level. Such action of M73101 was completely abolished by adrenalectomy, hypophysectomy, or the pretreatment with pentobarbital and morphine. In vitro experiment demonstrated that M73101 had no direct effect on adrenocortical function. These results indicate that the response to M73101 must be medicated through the release of ACTH from the adenohypophysis, which is probably due to the secretion of corticotropin releasing factor from the hypothalamus. M73101 at an oral dose of 200 kg/mg significantly decreased the volume of exudative fluid and the number of leucocytes in carrageenin-induced pleurisy of intact rats. However, the inhibitory action of this drug on cell mobilization decreased by adrenalectomy but not on exudative fluid, indicating that anti-inflammatory actions of M73101 may be due in part to pituitary adrenocortical stimulation. PMID- 6278122 TI - Further studies on the determinant role of brain level of pentobarbital for the development of acute hypnotic tolerance. AB - Controlling the brain level of pentobarbital and the duration of hypnosis at the initial treatment in relation to development of acute hypnotic tolerance was studied. Simultaneous treatment of bemegride or TRH with pentobarbital attenuated the hypnotic effect of pentobarbital in a dose dependent manner, but neither the brain level of pentobarbital nor the development of tolerance was modified by this treatment. The effect of TRH was further demonstrated in rats by concomitant intracarotidal infusion with pentobarbital maintaining the brain concentration of pentobarbital and also the duration of exposure of the brain to pentobarbital under a constant condition. On the other hand, THC significantly prolonged the hypnosis induced by pentobarbital but did not potentiate the effect of pentobarbital to develop acute tolerance. Thus, the brain level of pentobarbital at the initial treatment is the primary determinant for the development of acute tolerance and the duration of hypnosis is not the essential factor in this mechanism. PMID- 6278123 TI - Vitamin D: an update. PMID- 6278124 TI - Antitumor agents XLVII: The effects of bisbrusatolyl malonate on P-388 lymphocytic leukemia cell metabolism. AB - Bisbrusatolyl malonate, which was shown previously to be active against P-388 lymphocytic leukemia cell growth, was investigated for inhibitory effects on nucleic acid and protein synthesis. DNA and RNA synthesis as well as protein synthesis were markedly inhibited at 10,25, and 50 mu mole final concentrations in vitro. The major sites of inhibition of nucleic acid synthesis appeared to be DNA polymerase, messenger and transfer RNA polymerases, orotidine-5' monophosphate decarboxylase, phosphoribosyl pyrophosphate amino transferase, and dihydrofolate reductase. Moderate inhibition of nucleotide kinase activities and oxidative phosphorylation processes occurred after drug treatment. Cyclic adenosine monophosphate levels were reduced. Protein synthesis was inhibited during the elongation step of peptide synthesis. The data suggested that bisbrusatolyl malonate interfered with the peptide bond formation. However, the ongoing polypeptide synthesis must be completed before the drug can bind to the ribosome effectively. PMID- 6278125 TI - Effects of local anesthetic QX-314 on the membrane properties of hippocampal pyramidal neurons. AB - The quaternary lidocaine derivative QX-314 was applied internally to CA1 pyramidal neurons of the guinea-pig hippocampal slice preparation. This local anesthetic blocked both fast, Na+-dependent action potentials and the voltage dependent, non-inactivating Na+ conductance. Partially blocked Na+ spikes exhibited pronounced frequency-dependent depression at rates as low as 0.2 Hz. Ca++-dependent electrogenesis, synaptic potentials and glutamate-induced depolarizations were apparently unaffected even after large doses of QX-314. The results indicate that the cellular mechanisms of local anesthetics on central neuronal membranes are similar to those described for peripheral axons. The frequency-dependence of spike blockade may account for some of the effects of local anesthetics on the central nervous system in vivo. Additionally, the localized intracellular application of QX-314 may prove useful as a specific pharmacological tool in studies of central neurons. PMID- 6278126 TI - Lack of an opiate response to nitrous oxide in mice resistant to the activity stimulating effects of morphine. AB - Nitrous oxide, like morphine, stimulates locomotor activity in the CD-1, C-57 and C-3H mouse strains. This stimulation of locomotor activity is antagonized by the narcotic antagonist naltrexone (3 and 30 mg/kg). There are several strains (A/J and DBA/2J) of mice in which morphine does not stimulate locomotor activity. Mice resistant to this behavioral effect of morphine show a stimulation of locomotor activity in response to nitrous oxide which is not blocked by naltrexone. These results provide new data linking the effects of morphine and nitrous oxide. The opiate-like stimulation of activity produced by nitrous oxide may result from the release of endogenous opiate-like substances. Nitrous oxide causes a displacement of [3H]naloxone from brain opiate receptors labeled in vivo in the CD-1 mouse strain. In marked contrast to these results, nitrous oxide administration to the A/J and DBA/2J morphine-resistant strains failed to displace [3H]naloxone from opiate receptors labeled in vivo. Thus, variations in the response to nitrous oxide may not only be strain-dependent but possibly related to deficits in the response of or interaction with the endogenous opiate system. PMID- 6278127 TI - Naltrexone improves survival rate and cardiovascular function in canine hemorrhagic shock. AB - The possible involvement of opiate receptors in the cardiovascular depression associated with hypovolemic shock was investigated. Opiate receptor blockade with naltrexone increased mean arterial pressure, cardiac output, stroke volume and left ventricular contractility in dogs bled to a mean arterial pressure of 45 mm Hg. Naltrexone also increased survival rate. At high doses, naltrexone adversely affected cardiac performance which may outweigh its advantages of greater potency and putatively longer action than naloxone, at least in the dog. Similar actions with another opiate antagonist gives further proof for endogenous opiate involvement in the cardiovascular pathophysiology of hypovolemic shock. PMID- 6278128 TI - Maturation of sympathetic neurotransmission in the rat heart. VIII. Slowed development of noradrenergic synapses resulting from hypothyroidism. PMID- 6278129 TI - gamma-Aminobutyric acid (GABA) receptor stimulation. I. Neuropharmacological profiles of progabide (SL 76002) and SL 75102, with emphasis on their anticonvulsant spectra. AB - Progabide (4-([(4-chlorophenyl) (5-fluoro-2-hydroxyphenyl)-methylene]amino) butanamide) is a gamma-aminobutyric acid (GABA) receptor agonist which readily enters the brain. In the body, progabide is metabolized to three active metabolites: SL 75102, gabamide and GABA. Progabide and SL 75102 readily enter the brain and GABA and gabamide are also formed within this organ. Both progabide and SL 75102 exhibit a broad spectrum of anticonvulsant activities against seizures which involve GABA-mediated events (bicuculline, picrotoxinin and pentylenetetrazol) or which are apparently independent of GABAergic mechanisms (penicillin, strychnine, electroshock and audiogenic seizures). These data support the hypothesis that direct GABA receptor stimulation is an effective means of controlling convulsions of various origins. Progabide and SL 75102 have relatively minor secondary effects in comparison to commonly used antiepileptics. Myorelaxation occurs, but only at doses higher than the ED50 values in convulsant tests. Furthermore, these compounds are not sedative. Finally, these GABA agonists have a complex action in the extrapyramidal system. Anticonvulsant doses are antagonistic to dopamine receptor-mediated behaviors, whereas much lower doses seem to facilitate the effects of dopaminergic transmission. PMID- 6278130 TI - gamma-Aminobutyric acid (GABA) receptor stimulation. III. Effect of progabide (SL 76002) on norepinephrine, dopamine and 5-hydroxytryptamine turnover in rat brain areas. PMID- 6278131 TI - gamma-Aminobutyric acid (GABA) receptor stimulation. IV. Effect of progabide (SL 76002) and other GABAergic agents on acetylcholine turnover in rat brain areas. PMID- 6278132 TI - Degradation of exogenous enkephalin in the guinea-pig ileum: relative importance of aminopeptidase, enkephalinase and angiotensin converting enzyme activity. PMID- 6278133 TI - Enhanced angiotensin-mediated facilitation of adrenergic neurotransmission in spontaneously hypertensive rats. AB - The effect of angiotensin II on the pressor response of the mesenteric vascular bed to periarterial adrenergic nerve stimulation (PNS) was compared between spontaneously hypertensive rats (SHR) and normotensive Wistar Kyoto rats (WKY). Angiotensin II (1-20 ng/ml) caused a marked potentiation of the vasoconstrictor response to PNS (8 Hz, 2 msec, for 30 sec) in a concentration-dependent manner without significantly affecting the base-line perfusion pressure in either WKY or SHR. The facilitatory effect of angiotensin II was significantly greater in SHR than in WKY preparations. The effective concentration for the peptide in SHR was approximately one-third of that found in WKY. Angiotensin II also potentiated the pressor response to exogenously administered norepinephrine (NE, 50 ng) in both WKY and SHR. The degree of potentiation of the response to PNS was greater than that to NE in SHR, while this differential was found only at high concentrations of angiotensin II on the WKY. The facilitatory effect of angiotensin II on the pressor response to either PNS or NE was markedly reduced by [Sar1 Ile8]angiotensin II (200 ng/ml) in both WKY and SHR. Angiotensin II caused further potentiation of the vasoconstrictor response to PNS or NE infusion in the presence of cocaine (5 micrograms/ml), which was of a greater magnitude in SHR than in WKY. These results suggest that the presynaptic facilitatory modulation of adrenergic vascular neurotransmission, mediated by angiotensin II receptors, is enhanced in the perfused mesenteric vascular bed of SHR. PMID- 6278134 TI - Phenytoin antagonism of electrically induced maximal seizures in frogs and mice and effects on central nervous system levels of adenosine 3',5'-monophosphate and guanosine 3',5'-monophosphate. AB - Phenytoin antagonized the electroshock-induced increase in levels of cyclic adenosine 3',5'-monophosphate (cAMP) and cyclic guanosine 3',5'-monophosphate (cGMP) in cerebrum and cerebellum, respectively, from CF-1 mice. However, the effective dose range of phenytoin for significant reduction of the elevated levels of cAMP and cGMP was 2 to 5 times higher than that for prevention of tonic hindlimb extension in 95% of mice. The effective dose range of phenytoin for alteration of cyclic nucleotide levels was nearer to that for preventing tonic flexion and clonus; endpoints which are less relevant to anticonvulsant efficacy than is prevention of tonic hindlimb extension. Also , the greatest reduction in cyclic nucleotide levels occurred at a dose (100 mg/kg) which produced toxic signs in mice. Quaking mice (qk/qk), a mutant strain which exhibits spontaneous seizures, did not have abnormal levels of cAMP or cGMP in cerebrum or cerebellum, and a dose of phenytoin (15 mg/kg) which abolished all seizure activity did not alter levels of these cyclic nucleotides. In frogs, the electroshock-associated increase in levels of cAMP in the central nervous system was not altered by phenytoin even when the doses administered were up to twice the ED95 for prevention of tonic hindlimb extension. Because these data from mice and frogs show that the anticonvulsant effect of phenytoin is dissociated, by dose, from effects on central nervous system cyclic nucleotide levels, it is doubtful that the alteration of cyclic nucleotide levels is a mechanism by which phenytoin exerts its anticonvulsant effect PMID- 6278135 TI - Comparison of vanadate and ouabain effects on liver hemodynamics and bile production in the perfused rat liver. AB - Cumulative addition of vanadate or ouabain to rat liver perfusions giving perfusate concentrations up to about 500 microM revealed that both liver hemodynamics and bile production are influenced by these substances, but their ways of action differed markedly. Vanadate increased hepatic vascular resistance in a dose-dependent manner with an apparent Km of 30 microM. Not until vanadate concentrations in perfusate reached 60 to 70 microM did the hepatic oxygen consumption decrease significantly together with a decrease in bile flow. Ouabain at perfusate concentrations up to nearly 300 microM caused only a slight fall in perfusate flow, a dose-dependent slight fall in oxygen uptake and a dose dependent, marked increase in bile flow. Further addition of even small amounts of ouabain initiated a marked fall in perfusate flow, oxygen uptake and bile production and appeared to induce maldistribution of intrahepatic perfusate flow. The vasoconstrictive effect of vanadate was not influenced by alpha or beta blockers, atropine or blockers of Ca++-transport, whereas the effect of ouabain could be strongly reduced by phenoxybenzamine or verapamil. Vanadate-induced vasoconstriction may be caused by an inhibition of smooth muscle Ca++-adenosine triphosphatase and ouabain may induce vasoconstriction by inhibiton of smooth muscle Na, K-adenosine triphosphatase. The hepatic uptake and excretion of ouabain may explain the choleresis observed at small perfusate concentrations of ouabain. Inhibition of bile production at higher concentrations of ouabain and vanadate could be secondary to simultaneous changes in liver hemodynamics. PMID- 6278136 TI - Leukotrienes C4, D4 and E4: effects on human and guinea-pig cardiac preparations in vitro. AB - The effects of leukotrienes C4, D4 and E4 (LT C4, D4 and E4) were studied in isolated preparations of guinea-pig and human myocardium in order to assess their contribution to cardiac dysfunction associated with systemic anaphylaxis. LT C4, D4 and E4 all caused long-lasting and dose-related decreases in the contractile force and coronary flow rate of the isolated guinea-pig heart. The rank order of potency was LT D4 greater than C4 greater than E4. The effects of LT C4 and D4 were antagonized by the anti-slow-reacting-substance compound FPL 55712. The negative inotropic effect of LT is unlikely to be secondary to the concomitant reduction in coronary flow because: 1) the same reduction in coronary flow by angiotensin II resulted in a negligible decrease in contractility and 2) the negative inotropic effect of LT also occurred in the electrically paced, noncoronary perfused left atrium and right ventricular papillary muscle of the guinea pig and in pectinate muscles obtained from surgical specimens of human right atrial appendage. LT D4 potentiated the positive chronotropic effect of histamine, supporting the concept that functional interactions occur between the various mediators of immediate hypersensitivity. The cardiac effects of pure synthetic LT are similar to those previously obtained with crude slow-reacting substance of anaphylaxis indicating that the prolonged contractile failure associated with systemic anaphylaxis largely could be due to the negative inotropic effect of LT. Because LT are released in a variety of immunological and inflammatory reactions, their potent myocardial depressant effects may play a role in cardiac dysfunction associated with these reactions. PMID- 6278137 TI - Mobilization and removal of methylmercury in the dog during extracorporeal complexing hemodialysis with 2,3-dimercaptosuccinic acid (DMSA). PMID- 6278138 TI - Metabolism of acetaminophen by the isolated perfused kidney. AB - Acetaminophen (APAP) produces proximal tubular necrosis in the Fisher 344 rat. This lesion may result from the covalent binding of reactive intermediates of APAP to cellular macromolecules when glutathione (GSH) is sufficiently depleted. Experiments were designed to evaluate the ability of the kidney to convert APAP to reactive electrophilic metabolites capable of depleting renal GSH by quantifying GSH concentrations in isolated perfused kidneys perfused with APAP. Perfusion without APAP reduced (3 X 10(-5) -3 X 10(-5) M) to the perfusion medium further reduced renal GSH content. Treatment of rats with polybrominated biphenyls enhanced the ability of 3 X 10(-8) M APAP to deplete GSH. In contrast, treatment with piperonyl butoxide reduced the depletion of GSH produced by 3 X 10(-5) M APAP. At 3 X 10(-5) M APAP, the glucuronic acid, sulfate and the mercapturic acid conjugates were excreted by the isolate perfused kidneys. After treatment with polybrominated biphenyls, mercapturic acid excretion increased 4 fold, whereas the glucuronic acid and sulfate conjugate excretions were unaffected. These data suggest that the kidney can produce an electrophilic metabolite of APAP which can combine with and deplete renal GSH. An electrophilic metabolite of APAP produced by the kidney may initiate APAP induced renal necrosis. PMID- 6278139 TI - Effect of delta9-tetrahydrocannabinol on the synthesis of dopamine and norepinephrine in mouse brain synaptosomes. AB - The effects of delta 9-tetrahydrocannabinol (THC) on the conversion of [3H]tyrosine (TYR) to [3H]dopamine (DA) and [3H]norepinephrine (NE) by mouse brain synaptosomes were studied. The active uptake of TYR and its conversion to DA and NE were found to be concentrated in a synaptosomal subfraction prepared by using differential and density gradient centrifugation. Therefore, drug studies were performed by using the less pure P2 fraction. THC was administered in vitro by using a polyvinylpyrrolidone vehicle. DA synthesis was examined both in synaptosomes prepared from whole brain and from corpus striatum. THC produced increases in the conversion of TYR to DA in both preparations. Synthesis was stimulated maximally by 10 microns THC in both preparations. Higher concentrations produced either no further increase (whole brain) or a decrease in the synthesis of DA. NE synthesis was measured in synaptosomes prepared from whole brain and from brainstem. A concentration-dependent biphasic response was observed in both preparations. Peak increases in NE synthesis were observed with a 10 micron concentration of THC in synaptosomes from whole brain and a 3 micro concentration in the brainstem. THC also inhibited the active uptake of TYR into synaptosomes in all preparations studied. The lC50 in whole brain was 12.2 microns. The decrease in precursor uptake may be the cause of the decreased DA and NE synthesis observed at the higher concentrations of THC. The data from these studies suggest that delta 9-THC can have a direct effect upon catecholamine-containing neurons in the brain. PMID- 6278140 TI - The effect of ionic environment in specific FSH binding to plasma membrane receptor. AB - Studies were undertaken to define the effects of ionic strength and ion selectivity on specific binding of 125I-hFSH to both its particulate and Triton X 100 solubilized calf testis plasma membrane receptor. Plasma membrane vesicles migrated with a negative charge on free flow electrophoresis. There was no evidence of formation of inside-out vesicles. Specific FSH binding to the plasma membrane vesicles with both monovalent and divalent cations, used to reduce charge repulsion between the negatively charged FSH molecule and membrane vesicles, revealed similar binding maxima at an ionic strength range of 0.03 to 0.05. However, Mg2+ consistently resulted in 25-30% greater specific binding than with any other cation. That increased FSH binding suggested unmasking of specific FSH plasma membrane binding sites. However, specific FSH binding to solubilized plasma membrane vesicles was unaffected by ionic strength or ion selectivity. Enzymatic removal of sialic acid from plasma membrane vesicles significantly enhanced specific FSH binding. In previous studies, we had observed that sialic acid was not exposed within the FSH binding site. Consequently, enhanced FSH binding associated with desialylation of plasma membrane vesicles probably reflected modification of parareceptor sites which affected the conformation of the specific FSH receptor. PMID- 6278141 TI - Epstein-Barr virus receptor expression is correlated to cell cycle phase. AB - Epstein-Barr virus (EBV) receptor positive cells absorbed FITC-conjugated virions and were subsequently stained for DNA content with propidium iodide. These cells were analyzed by flow cytometry to determine the relationship between cell cycle and EBV receptor (EBVR) expression on single cells. EBVR was present throughout the entire cell cycle. The level of expression, however, varied; increasing in G1 and G2/M while remaining approximately constant in S phase. Analysis of receptor density in terms of cell size demonstrated that smaller cells, in early G1, have a greater density of receptors per unit membrane than larger G2/M and S phase cells. PMID- 6278142 TI - Release of erythropoietin from macrophages mediated by phagocytosis of crystalline silica. AB - The hormone erythropoietin (Ep), which regulates erythropoiesis, can be shown to be released extrarenally from macrophages of the spleen, bone marrow, peritoneal exudate, lung, liver, and fetal liver when suspensions of these cells are preincubated with crystalline silica. The cell supernatants are assayed for Ep activity in the erythroid colony-forming technique using 12- 13-day fresh or cryopreserved fetal liver cell suspensions as a source of Ep-sensitive CFU-E target cells. This report describes the optimal conditions required for release of Ep from silica-treated spleen macrophages. The requirement for phagocytosis of silica in mediating Ep release is shown by four different methods, namely, temperature dependency, inhibition of phagocytosis by Cytochalasin B, the presence of calcium in the medium, and the fact that silica particles coated with serum or poly-2-vinylpyridine-N-oxide can decrease Ep release to control levels. In addition, the reduction to background Ep levels when reduced glutathione, alpha tocopherol (vitamin E), and alpha-thioglycerol are added either separately or in combination with silica-treated macrophages is discussed in terms of a possible mechanism of silica action. PMID- 6278143 TI - Serum-dependent and -independent effects of bacterial lipopolysaccharides on human neutrophil oxidative capacity in vitro. PMID- 6278144 TI - Evidence of a T-cell-mediated cytotoxic response to parainfluenza virus type 3 pneumonia in hamsters. PMID- 6278145 TI - The effect of cytochalasin B on the superoxide production by alveolar macrophages obtained from normal rabbit lungs. AB - Resident alveolar macrophages and lung lavage fluids were obtained from normal rabbit lungs. Superoxide production by alveolar macrophages exposed to lung lavage fluids and cytochalasin B was measured by superoxide dismutase-inhibitable nitroblue tetrazolium (NBT) reduction. Glycogen-elicited peritoneal macrophages were used as a control. Cytochalasin B, as well as lung lavage fluids, enhanced superoxide production by resting alveolar macrophages. The cytochalasin B-induced superoxide production was associated with enhanced attachment to glass and with remarkable alterations of the cell surface morphology, probably relating to interference with the microfilament functions of cells. On the other hand, glycogen-elicited peritoneal macrophages showed only a slight production of superoxide when exposed to the same stimulants. PMID- 6278146 TI - Autosomal dominant hypoparathyroidism: a proband with concurrent nephrogenic diabetes insipidus. AB - In this paper we report an extended family with well documented autosomal dominant hypoparathyroidism which was ascertained through a proband with coincident nephrogenic diabetes insipidus. Clinical findings were limited to a slight decrease in overall stature and to clinical signs of hypocalcaemia. Intelligence was normal and two patients were asymptomatic. Published reports have established that autosomal dominant, autosomal recessive, and sex linked recessive familial isolated hypoparathyroidism exist. However, in almost half the reported families an X linked dominant aetiology cannot be excluded and, at present, clinical criteria provide only minimal aid in distinguishing between the different genetic types. There remains a need for detailed documentation of further families were the pattern of inheritance is clear. PMID- 6278147 TI - A case of the orocraniodigital (Juberg-Hayward) syndrome. AB - A female with the orocraniodigital (Juberg-Hayward) syndrome is described in whom, in addition to bilateral cleft lip and palate, mild microcephaly, and anomalous thumbs and toes, there was absence of the pituitary fossa and a more widespread skeletal dysplasia. PMID- 6278148 TI - Protective action of calcium channel antagonist agents against ventricular fibrillation in the isolated perfused rat heart. PMID- 6278149 TI - Distribution and properties of cAMP-dependent protein kinase isozymes in the myocardium of spontaneously hypertensive rat. PMID- 6278150 TI - Inorganic phosphate: its effects on Ca exchange and compartmentalization in cultured heart cells. PMID- 6278151 TI - Poly (ADP-ribose) polymerase activity of aortic nuclei from swine on hyperlipemic diet. PMID- 6278152 TI - Estimating evolutionary distance from restriction maps of mitochondrial DNA with arbitrary G+C content. AB - We develop a mathematical model for estimating evolutionary distance from restriction enzyme maps, which incorporate non-uniformity of the rate of base substitution into the theory and allows for an arbitrary G+C content at equilibrium. When the G+C content differs significantly from 1/2, the traditional model of base changes can introduce a systematic bias which depends upon the base composition of the restriction site. In addition, the accuracy of estimated evolutionary distance depends heavily upon the choice of restriction enzyme in that the expected number of sites is also affected. Monte Carlo experiments are conducted to check the validity of the present theoretical treatment and from which we draw several cautionary notes on estimation. An application is made to the available data on restriction enzyme maps of human mitochondrial DNA where the G+C content is approximately 1/3. PMID- 6278153 TI - Effects of polychlorinated biphenyls on the development of intestinal and serum marker enzymes. AB - The effects of polychlorinated biphenyls (PCBs) on the development of several intestinal and serum marker enzymes have been studied. The three congeners 4 monochloro (1-CB), 3, 4, 3',4'-tetrachloro (4-CB), and 2, 4, 5, 2', 4, 5' hexachloro (6-CB) biphenyl were administered orally to pregnant rats on d 8, 11, 13, 15, and 18 or gestation. 1-CB and 6-CB were intubated at doses of 30 mg/kg.d (total dose, 150, mg/kg) and 4-CB was administered at 3 mg/kg-d (total dose, 15 mg/kg). Levels of intestinal alkaline phosphatase, monoamine oxidase, and Na+, K+ adenosin-5'-etriphosphatase and levels of serum alkaline phosphatase, sorbitol dehydrogenase, and beta-hydroxybutyrate dehydrogenase were measured in the dams after weaning and in their offspring at -1, 6, 20, and 55 d of age. Intestinal alkaline phosphatase activity was elevated at the later postnatal stages in the 1 CB group and depressed at 55 d in the 4-CB group, whereas serum alkaline phosphatase levels were markedly depressed prenatally and postnatally in the 4-CB and 6-CB groups, respectively, Intestinal monoamine oxidase levels were markedly increased in the 6-CB group at -1, 6, and 20 d of age and significantly depressed in the 4-CB animals at -1 and 55 d of age. There was an increase in monoamine oxidase activity in the 4-CB group at 6 d. The 1-CB group exhibited depression of monoamine oxidase levels at 6 and elevation at 20 and 55 d. Intestinal Na+, K+ ATPase levels were elevated throughout development in the 1-CB animals and at -1 and 6 d in the 4-CB group. The 6-CB animals showed elevated levels of Na+, K+ ATPase only at 6 d. Serum beta-hydroxybutyrate dehydrogenase and sorbitol dehydrogenase were induced prenatally in the 4-CB animals but enzyme activities decreased to normal by 55 d of age. Significant depression of activity was evident in the 1-CB and 6-CB groups at -1 d in both enzymes. Dams in the 1-CB group showed significant changes in intestinal monoamine oxidase, serum sorbitol dehydrogenase, and serum alkaline phosphatase. Serum levels were elevated in the 4-CB group. Activities of intestinal enzymes remained unchanged in the 4-CB group. All maternal enzyme levels monitored were not significantly changed in the 6-CB group. PMID- 6278154 TI - Cardiac and hepatic effects of pre- and postnatal exposure to polybrominated biphenyls in rats. AB - Body weight gain and hepatic concentrations of vitamin A were reduced in Sprague Dawley rats by pre- and postnatal exposure to 100 ppm polybrominated biphenyls (PBBs). The ratio of liver weight to body weight, activity of hepatic delta aminolevulinic acid (ALA) synthetase, and urinary excretion of uro- and coproporphyrins were increased by PBBs. Treatment with PBBs also increased the left atrial inotropic response to calcium. However, PBBs had no effect on development of the adrenergic neuronal transport system in heart, left atrial baselike peak tension, or inotropic response to ouabain. Thus PBBs retarded body weight gain and produced a variety of alterations in liver, but had little effect on cardiac contractile function. PMID- 6278155 TI - Nonrespiratory metabolic function and morphology of lung following exposure to polybrominated biphenyls in rats. AB - Exposure to polybrominated biphenyls (PBBs) resulted in increased activity of microsomal arylhydrocarbon hydroxylase and ethoxyresorufin-O-deethylase in rat lung. Clearance of 5-hydroxytryptamine (5-HT) and angiotensin 1 by perfused lungs was decreased by PBBs. However, PBBs had no effect on the activity of epoxide hydrolase, monoamine oxidase, or angiotensin-converting enzyme in lung. The only histopathologic change detected in lungs from PBB-treated rats was an increase in alveolar type II cell lamellar bodies. Selective accumulation of certain PBB congeners by lung was not observed in this investigation. PMID- 6278156 TI - Effect of morphine on catecholamine - stimulated cyclic AMP production in cortex slices from rats and mice. AB - Morphine (10 micro M) blocked noradrenaline-stimulated cyclic AMP production in slices of cerebral cortex from normal rats but not in slices from rats pretreated with 6-hydroxydopamine (6-OHDA). In contrast, morphine failed to prevent noradrenaline-stimulated cyclic AMP production in mouse cortex slices. Levorphanol weakly antagonized the rise in cyclic AMP produced by noradrenaline in both normal and 6-OHDA-treated mouse cortex. Morphine had no effect on the adrenaline-stimulated cyclic AMP accumulation in mouse cortex but it entirely prevented the rise in cyclic AMP produced by isoprenaline. This effect was no observed in brain slices from 6-OHDA-treated mice. It is concluded that in slices of rat cortex, morphine stimulates postulated presynaptic, 6-OHDA-sensitive, opiate receptors associated with noradrenergic nerve terminals. These opiate receptors alter postsynaptic alpha - and beta - adrenoceptor activity. In the mouse, morphine appears to stimulate presynaptic opiate receptors that modify exclusively beta - adrenoceptor-mediated cyclic AMP production. PMID- 6278157 TI - Differential effects of morphine withdrawal on cerebral beta 1- and beta 2 adrenergic receptors. AB - Effect of morphine dependence and its withdrawal on the 3H-dihydroalprenolol (3H DHA) binding for beta -adrenergic receptors, beta 1 and beta 2, was examined by a computerized analysis of biphasic Hofstee plots. The relative density of beta 1 and beta 2 receptors in the rat cerebral cortex was found to be approximately 70% and 30%, respectively. In rats rendered dependent on morphine by a subcutaneous implantation of a morphine pellet, the 3H-DHA binding to beta 1 and beta 2 receptors was not altered. During the stage of withdrawal induced by administration of naloxone, however, the 3H-DHA binding to the cerebral particulate fractions was increased, and this increase was due to the increased binding sites in beta 1 and beta 2 receptors. On the other hand, the apparent affinities of beta and beta 2 for atenolol and salbutamol, selective antagonists for beta 1- and beta 2-adrenergic receptors, respectively, were not altered under these experimental conditions. These results suggest that an abrupt increase in cerebral beta 1-receptor binding sites occurs at morphine withdrawal, and the occurrence of such a super-sensitivity in cerebral beta 1 receptor may be involved in the exhibition and/or maintenance of the abstinence syndrome in morphine-dependent subjects. PMID- 6278158 TI - Exaggerated cyclic AMP accumulation and glial cell reaction in the cerebellum during Purkinje cell degeneration in pcd mutant mice. AB - The Purkinje cell degeneration mutant (pcd) is characterized by a complete loss of cerebellar Purkinje cells. Norepinephrine causes an accumulation of cyclic AMP in the cerebellum of pcd that is far greater than in normal mice. Experiments were conducted 1) to correlated changes in the cyclic-nucleotide response with a histologic examination of the cerebellum during neuronal loss and 2) to examine the role of cyclic AMP catabolism and adenosine receptor interactions in the phenomenon. The greatest elevation in cyclic AMP occurred between 30 and 128 days of age when a severe astrocytic response was demonstrated throughout the cerebellar cortex. Purkinje cells had degenerated by 45 days of age. Norepinephrine elicited a smaller increase in cyclic AMP from 155-day-old mice than at earlier ages, and the response continued to decrease with age; at 270 days, equal accumulation, and at 365 days. lower accumulation of cyclic AMP was detected in pcd cerebella. During this time, the Purkinje cell debris had been removed, the granule cell layer was depleted of granule cells, and the molecular layer was deprived of a large number of parallel fibers. However, although phagocytosis of neuronal debris was completed, large numbers of astrocytic processes were still seen in the neuropil. Biochemical experiments in vitro established that the exaggerated accumulation of cyclic AMP in the presence of norepinephrine was not due to lower catabolism of cyclic AMP, a synergistic interaction with adenosine, or a result of lower protein in the pcd cerebellum. The correlates of heightened norepinephrine-stimulated accumulation of cyclic AMP with neuronal loss and the glial cell reaction might indicate that cyclic nucleotides play a role in controlling some glial cell functions, ie, proliferation, migration, and phagocytosis. PMID- 6278159 TI - Effect of exogenous adenosine 3', 5'-cyclic monophosphate (cAMP) on the ultrastructure of neonatal rat hepatocytes in primary tissue culture. A stereological investigation. AB - Daily administration of exogenous cAMP to neonatal rat liver primary cultures caused a decrease of the total cellular volume, membrane space and SER coupled with a marked increase in the complement of RER, free ribosomes and polysomes and in the volume of nuclei after 3 days in hepatocytes; moreover the Golgi apparatus appeared to have undergone hypertrophy and the microtubules to have formed noticeable bundles. The present results support the view that the functioning of the RNA and protein synthesizing machinery of neonatal rat hepatocytes is stimulated by exogenous cAMP. PMID- 6278160 TI - Ultrastructural changes in the adrenocortical cells of post-hatching chicks (Gallus domesticus) following ACTH treatment. AB - Adrenal glands from control and experimental young chicks (Gallus domesticus) were collected and processed for electron microscopy. Control interrenal cells displayed ultrastructural characteristics common to other steroid-secreting cells. Blood sinusoids always remained open and endothelial cells of the blood capillary were well preserved. Under the influence of ACTH, lipid translucent droplets disappeared and were replaced by a few dense lipid droplets. These latter structures and lysosomes were often seen close to Golgi complexes. As the lipid translucent droplets increased in size, they became closely associated with mitochondria, smooth endoplasmic reticulum, and lysosomes thus suggesting a structural-functional relationship. ACTH caused a depletion of lipid droplets and an increase in the number of mitochondria, smooth endoplasmic reticulum, lysosomes, and desmosomes. Mitochondria generally showed a change in matrix density and cristae formation. Some were found to be fragmenting into smaller units whereas others were seen enclosing cytoplasmic matrix. Smooth endoplasmic reticulum was predominantly of the swollen type and lysosomes were primarily concentrated at the plasma membrane. The nucleolus showed a predominance of dense stippled materials as opposed to light stippled material. All these changes suggest that ACTH did affect the cortical cells of 17 day old post-hatching chicks. PMID- 6278161 TI - Axo-dendritic abnormalities in a case of juvenile neuronal storage disease. AB - Axonal enlargements (meganeurites) and dendritic degeneration were found in the pyramidal neurons from the frontal cortex of a patient with juvenile neuronal storage disease studied with the aid of ultrastructural and Golgi methods. Meganeurites appeared as enlargements at the axon hillock-initial segment area and were similar to those described in other neuronal diseases; dendritic degeneration was heavy and appeared throughout the cell domain. These neuropile changes can be related to the precocious mental abnormalities observed in this group of diseases which depend on a storage phenomenon due to a lysosomal enzyme deficiency. PMID- 6278162 TI - Ultrastructural alterations of the lymphocytes from a patient with Reye's syndrome. AB - The ultrastructural alterations of the peripheral blood mononuclear cells of a patient with Reye's syndrome are reported. The involved cells were lymphocytes which showed marked nuclear changes such as irregular or highly convoluted nuclei, multilobulated nuclei, presence of scanty chromatin, and large nucleoli. Changes of the cytoplasmic organelles involved the mitochondria and dense bodies with well defined surrounding membrane appeared. The possibility that these findings indicate a viral etiology of the disease is discussed. PMID- 6278163 TI - Clinicopathological Conference: Sudden anuria and renal failure in an elderly man. PMID- 6278164 TI - Testicular malacoplakia. AB - We report the twentieth case in the world literature of testicular malacoplakia. This lesion occurs mainly in middle-aged men, appearing clinically as epididymo orchitis or enlargement of the testicle with fibrous consistency and some soft areas. Malacoplakia is now considered a nonspecific inflammatory granulomatous lesion subsequent to an infection by gram-negative bacteria. In our case Escherichia coli was cultured in the urine and testicular tissue. The ultrastructural study shows the histiocytic nature of the cellular infiltrate as well as the morphology of Michaelis-Gutmann bodies. Bacilliform structures were not found. The cellular and humoral immunological studies did not show alterations. PMID- 6278165 TI - Multimodal treatment of advanced adult Wilms tumor. AB - We report 2 cases of advanced adult Wilms tumor that were treated with surgery, radiation and chemotherapy. The first patient had relapse of a Wilms tumor in the liver 2 years after nephrectomy. Combination chemotherapy, consisting of actinomycin D and vincristine, radiation therapy and final resection of the liver metastasis were successful and the patient has been free of disease for 4 years. The second patient had undergone transcatheter embolization of the renal artery elsewhere with the tentative diagnosis of an inoperable renal cell carcinoma metastatic to both lungs. A left renal tumor, weighing 4,500 gm., and a tumor thrombus in the vena cava extending to the right atrium were removed, and histologically diagnosed as a Wilms tumor. Subsequent chemotherapy and radiotherapy resulted in complete disappearance of the lung metastases. We conclude that multimodal treatment, namely a well timed combination of surgery, chemotherapy and radiotherapy, could potentially eradicate the disease even at an advanced stage. PMID- 6278166 TI - Advances continue in sickle cell disease. PMID- 6278167 TI - Cold urticaria in infectious mononucleosis. PMID- 6278168 TI - Fatal lymphohistiocytic proliferation owing to Epstein-Barr virus. PMID- 6278169 TI - The clinical relevance of 'CSF viral culture'. A two-year experience with aseptic meningitis in Rochester, NY. AB - The clinical relevance of CSF viral cultures was evaluated by reviewing the records of 390 patients whose CSF was cultured for virus during a two-year period. The diagnoses at hospital discharge were aseptic meningitis, meningoencephalitis, or both in 111 patients, and enterovirus was isolated from the CSF or other test specimens in 46 patients (41%). The diagnosis or management of nearly one half of the patients from whom enterovirus was isolated was directly influenced by this information. Hospitalization and the unnecessary use of antibiotics were shortened by at least 70 days. Enterovirus was the only virus isolated from the CSF during the study period. The CSF was more likely positive for an enterovirus if it was drawn from a young patient with aseptic meningitis during the summer of fall months. The clinical data obtained from this study are discussed and compared with national statistics. PMID- 6278170 TI - Human tissue burdens of halogenated aromatic chemicals in Michigan. AB - A cross section of the population of Michigan was studied in 1978 to evaluate residual burdens of polybrominated biphenyls (PBBs), a flame-retardant chemical that had contaminated the state's food supply five years earlier. Levels of PBB were measured in serum specimens in 1,681 people and in adipose samples in 844. Ninety-seven percent of the adipose samples had detectable PBB concentrations. Levels of PBB were highest in that part of the state where meat and dairy products had been most contaminated and were lowest in the upper peninsula, farthest from the source. Levels for the rest of the state were in between. Polychlorinated biphenyls (PCB) and 2,2'-bis(p-chlorophenyl)-1, 1 dichloroethylene (p,p'-DDE), a dichlorodiphenyltrichloroethane (DDT) residue, were also measured, and variations of PBB, PCB, and p,p'-DDE levels with age and sex were investigated. PMID- 6278171 TI - Tissue burdens of toxic pollutants. PMID- 6278172 TI - Nuclear magnetic resonance imaging. A promising technique. PMID- 6278173 TI - Effects of angiotensin I converting enzyme inhibitor (SQ 14,225) on the responses of blood pressure and steroid hormone to angiotensin II and ACTH infusion in hypertensive subjects. AB - The effects of the converting enzyme inhibitor, SQ 14,225, on the renin angiotensin system, adrenal function and blood pressure were investigated in 14 hypertensive patients, i.e., 10 with essential hypertension (EH) and 4 with renovascular hypertension (RVH). The mean blood pressure (MBP) and plasma aldosterone showed significant decreases in the EH with normal renin (NR) group and in the RVH group but no significant changes in the EH with low renin (LR) group. Plasma renin activity (PRA) increased significantly in the EH and NR group and in the RVH group but showed no significant change in the EH with LR group. Significant correlations were found between the fall in MBP after SQ 14,225 treatment and the pretreatment levels of PRA or plasma aldosterone. In an ACTH infusion study, the response of plasma aldosterone to ACTH revealed significant decreases after SQ 14,225 administration. In an angiotensin II (A II) infusion study, the response of plasma aldosterone was unchanged after SQ 14,225 administration. However, the pressor responses to A II infusion with SQ 14,225 were significantly higher than those without SQ 14,225. From these findings, it is concluded that the antihypertensive mechanism of SQ 14,225 may be due mainly to the decrease in levels of endogenous A II and that the reduction in plasma aldosterone after SQ 14,225 were significantly higher than those without SQ 14,225. From these findings, it is concluded that the antihypertensive mechanism of SQ 14,225 administration may be due to reduction of endogenous A II levels by converting enzyme inhibition. PMID- 6278174 TI - [Comparison of antibacterial activity of cefmenoxime with other cephalosporins against clinically isolated bacteria (author's transl)]. AB - We examined the antibacterial activity of MX in comparison with those of other CEPs, using aerobic Gram-positive cocci, aerobic Gram-negative bacilli and anaerobic bacteria, 870 strains in total, all isolated from clinical specimens, in 1979 and 1980. Against Streptococcus, CMX showed superior antibacterial activity than those of CFX, CMZ, CXM and CTM. Against H. influenzae, CMX also showed superior antibacterial activity than those of CFX, CMX, CXM, CTM and CEZ. ABPC-and PIPC-resistant strains were sensitive to CMX. CTX, CPZ and CZX also showed antibacterial activities equivalent to that of CMX. Against enteric bacteria, E. coli, Klebsiella, E. cloacae, Serratia, C. freundii and Proteus, CMX showed superior antibacterial activity than those of CFX, CMZ, CXM, CTM and CEZ. Especially, against E. coli, Klebsiella, P. mirabilis, P. rettgeri and P. inconstans, CMX showed strong antibacterial activity. As to non-fermentation bacteria, CMX's antibacterial activity was relatively weak except P. putrefaciens, Alcaligenes and Comamonas. However, it was superior than that of CEZ. In comparison with other CEPs, the strength of CMX varied according to the kinds of bacteria. As to anaerobic bacteria, CMX showed strong antibacterial activity against Peptococcus, Peptostreptococcus, Lactobacillus, Propionibacterium, C. perfringens, Veillonella and Fusobacterium. However, its antibacterial activity against Bacteroides was similar to those of other CEPs. PMID- 6278175 TI - [A fundamental and clinical study of ceftizoxime in the field of obstetrics and gynecology (author's transl)]. AB - A fundamental and clinical study of ceftizoxime (CZX), a new cephalosporin antibiotic, has brought about the following results. 1. The antibacterial activity of CZX against Bacteriodes fragilis and B. thetaiotaomicron was superior to that of CEZ, but inferior to that of CFX. CZX was the most active of the 3 drugs against B. distasonis, and was as active as CFX against B. vulgatus. The MICs of CZX against Peptococcus, Peptostreptococcus and anaerobic Streptococcus were 1.56 microgram/ml or lower. CEZ was more active than CZX against these anaerobic cocci. 2. The concentrations of CZX in female genital organs and retroperitoneal dead space exudate after drip infusion of 1 g were high enough to fulfill the MICs of many bacterial isolates. 3. CZX administered intravenously to 9 patients in daily doses of 1 approximately 8 g for 4 approximately 11 days was 100% effective. Pathogenic bacteria in 6 of the patients were eradicated in 4 and partially eradicated in 1. The other patient had no bacteriological examination after therapy. 4. No adverse reaction was observed. From above results it is concluded that CZX must be one of the most effective antibiotics for the treatment of gynecological infections. PMID- 6278176 TI - [Fundamental studies on excretion of pivmecillinam in bile (author's transl)]. AB - Excretion of pivmecillinam (PMPC) in bile was investigated using normal and hepatic disorder (intramuscular injection of carbon tetrachloride, 0.5 g/kg twice) rabbits. Bile and blood were collected hourly (0.5 approximately 8 hours) and both concentrations and rates between both (passage), and excretion rates were investigated. Ampicillin (AMPC) was chosen as a control drug. 1. When PMPC was given orally to the normal rabbit in a dose of 200 mg/kg, a bile concentration after 0.5 hour was 138.08 microgram/ml and a peak value was attained. The rate to plasma concentration was 22.67. An excretion rate was 0.224%. When PMPC was given orally to the hepatic disorder rabbit in a dose of 200 mg/kg, a bile concentration after 0.5 hour was 26.67 microgram/ml. A peak value of 28.87 microgram/ml was attained after 4 hours. Their rates to plasma concentrations were 13.27 and 8.02, respectively. An excretion rate was 0.058%. Bile and plasma concentrations, rates of bile to plasma and excretion rates of the hepatic disorder group were lower than those of the normal group. Disturbances of hepatic functions were checked by biochemical tests, i.e. GOT, and so on. 3. When ABPC as a control drug was given orally to the rabbit in a dose of 200 mg/kg, plasma concentrations of hepatic disorder group were higher than those of the normal group, but rates of bile to plasma concentrations and excretion rates of the hepatic disorder group were lower than those of the normal group. 4. Bile concentrations and excretion of PMPC were significantly higher than those of ABPC in the normal group. In the hepatic disorder group, however, no statistical difference was observed between them. PMID- 6278177 TI - [Laboratory and clinical studies on ceftizoxime in otorhinolaryngological field (author's transl)]. AB - From the laboratory and clinical studies on ceftizoxime (CZX), the following results were obtained 1) CZX was compared with cefazolin (CEZ) for in vitro activity against 6 standard strains and clinically isolated strains of Staphylococcus aureus (19 strains), Staphylococcus epidermidis (14), Proteus sp. (17), Escherichia coli (3), Klebsiella sp. (3) and Pseudomonas aeruginosa (13). While somewhat less active against Gram-positive cocci than CEZ, CZX was far more active than CEZ against Gram-negative bacilli. 2) The time course of mean serum CZX levels in 2 patients given a single intravenous dose of 10 mg/kg was as follows: 24.9 microgram/ml at 15 minutes, 18.5 microgram/ml at 30 minutes, 13.2 microgram/ml at 1 hour, 6.4 microgram/ml at 2 hours and 2.7 microgram/ml at 4 hours. The mean serum half life was 1.36 hours. The mean tonsil concentrations of CZX 30 minutes after a single intravenous dose of 0.5 g and 1.0 g were 5.9 microgram/g and 9.6 microgram/g. respectively, with the ratio to the serum concentration of 0.33 and 0.32. 3) CZX was given to 28 patients with ear, nose, and throat infections, and overall rate of effectiveness was 92.3%. No clinical side effects were observed. Changes in laboratory test findings included slightly elevated GOT and/or GPT in 3 cases. PMID- 6278178 TI - [Remarkable blood pressure change cycle seen in extra-adrenal pheochromocytoma]. PMID- 6278179 TI - Absence of antibodies against SV5, measles and herpes simplex in indoor colony bred cynomolgus monkeys. AB - One hundred and eighty-one cynomolgus monkeys born and raised at Tsukuba Primate Center for Medical Science (TPC) were serologically examined for the presence of antibodies against simian virus 5, measles virus and herpes simplex virus. All of them were virtually free from infections with these viruses, whereas wild originated breeders kept at TPC were highly positive for these antibodies. These results can be regarded as demonstrating that an indoor breeding system such as TPC's is useful and valuable for producing nonhuman primates of good quality being completely free from those viral infections. PMID- 6278180 TI - [Inhibition of angioten converting enzyme by phosphryl derivatives in rats (author's transl)]. PMID- 6278181 TI - [Effect of sodium intake on the brain and the aortic angiotensin-converting enzyme activity in spontaneously hypertensive rat (author's transl)]. PMID- 6278182 TI - Effect of hypothyroid status on adenosine 3',5'-mono-phosphate-dependent protein kinase of skeletal, heart and diaphragm muscle of rats. AB - Properties of cyclic AMP-dependent protein kinases from skeletal, heart and diaphragm muscles of hypothyroid rats were compared. Increased enzyme activity was observed in skeletal muscle from hypothyroid rats after DEAE-cellulose chromatography. Changes in isozyme distribution were also shown in the hypothyroid status. The elution profile on DEAE-cellulose suggested a possible translocation of the enzyme from the particulate to the soluble fraction in the heart of hypothyroid rats. The turnover rate of the enzyme protein decreased in the skeletal muscle of hypothyroid rats, but the other two organs showed no change even in the hypothyroid status. The activity of heat-stable protein kinase inhibitor increased in the skeletal and the diaphragm muscles in hypothyroid rats, whereas the activity in the heart decreased in the hypothyroid status. These data suggested the possibility that changes in enzyme properties, modification of isozyme distribution, and changes in modulator activity might account for the modulation of muscle function in hypothyroid rats. PMID- 6278183 TI - Effects of guanabenz on the adrenergic mechanism in rabbit arterial strips. AB - Effects of guanabenz, a hypotensive agent, on the adrenergic mechanism were studied in isolated rabbit thoracic aorta and pulmonary artery and findings were compared with data obtained with clonidine and guanethidine. Guanabenz in concentrations higher than 10(-6) M produced weak contractions which were attenuated by phentolamine or yohimbine. Such concentrations of guanabenz competitively inhibited the contractile response to noradrenaline, but did not attenuate the response to tyramine. In concentrations ranging from 10(-8) to 10( 7) M, guanabenz attenuated the contraction and the increase of 3H-efflux in response to transmural electrical stimulation of the pulmonary artery preincubated with 3H-noradrenaline. Phentolamine or yohimbine effectively blocked these inhibitory effects of guanabenz. Such effects of guanabenz were similar to those of clonidine and dissimilar to those of guanethidine. These results indicate that guanabenz acts on presynaptic and postsynaptic alpha receptors of the peripheral blood vessels, as in the case of clonidine and that the potency was almost the same as clonidine. PMID- 6278184 TI - Plasma cyclic AMP in the pethidine-dependent rat. PMID- 6278185 TI - Partial agonistic action of morphine in the rat vas deferens. AB - Effects of morphine on the force of contraction of rat vas deferens were investigated. Morphine and beta-endorphin decreased the electrically evoked twitch tension, in a dose dependent manner. The inhibitory effect of morphine, however, was much weaker than that of beta-endorphin. These effects of both morphine and beta-endorphin were completely antagonized by naloxone. In the presence of 30 microM morphine, the dose-response curve of beta-endorphin shifted to the right by about 10-fold. Moreover, morphine partly reversed the contraction depressed by 0.3 microM beta-endorphin, in a dose dependent manner. These findings suggest that morphine acts as a partial agonist on the rat vas deferens. Marked tolerance to beta-endorphin and change in the antagonist potency of morphine were not observed in the vas deferens isolated from morphine-dependent rats. PMID- 6278186 TI - Aftercare concerns of rape victims. PMID- 6278187 TI - Human lymphoma-lymphoma hybrids and lymphoma-leukemia hybrids. I. Isolation, characterization, cell surface markers, and B-cell markers. AB - Four new somatic cell hybrids were obtained by fusion of various Burkitt's lymphoma (BL)-derived cell lines that had different selective markers: Raji-P3HR 1, Daudi-Raji, and a P3HR-1-P3HR-1 "autohybrid" derived from two P3HR-1 sublines. In addition, a hybrid was obtained between the Daudi (BL) line and the human leukemia cell line K562. The hybrids were extensively characterized by means of chromosome, isozyme, and HLA surface markers. The phenotypic differences between the parent cell lines allowed some conclusions with respect to the expression of latent Epstein-Barr virus (EBV) genomes, C3 and EBV receptors, and of immunoglobulin and beta 2-microglobulin-HLA expression as well as the influence of the leukemia cell (K562) genome on B-cell properties in the Daudi-K562 hybrid. B-cell and differentiated markers of these hybrids were characterized. High-level expression dominated for the marker C3 and EBV receptors, which showed a good correlation coefficient of 0.84, as was true for Fc receptors and surface immunoglobulin. The Daudi-K562 hybrid showed loss of all B-cell markers but retention of the leukemia cell markers (e.g., hemoglobin synthesis). PMID- 6278188 TI - Human lymphoma-lymphoma hybrids and lymphoma-leukemia hybrids. II. Epstein-Barr virus induction patterns. AB - In addition to the spontaneous expression of markers of the early and late phases of the viral cycle [Epstein-Barr virus (EBV), early viral cycle antigens (EA), and viral capsid antigen (VCA)] by a Panel of hybrid cell lines derived by fusion of human hematopoietic cell lines, the induction of these markers by three chemical inducers [5-iodo-2'-deoxyuridine, the tumor promoter 12-O tetradecanoylphorbol 13-acetate (TPA), and sodium butyrate] were analyzed. A variant of the prototype producer of lytic EBV, P3HR-1 (PICAT) was observed to show consistent low spontaneous and induced production of EA and VCA as compared to the P3HR-1 line from which it was derived. An "autohybrid" (PICATPO-1) between these two lines showed a low production of VCA. Hybrids between Raji and P3HR-1 sublines (RUD-PICAT-1 and RUDPUT-2) showed reduced expression of EBV antigens. Both hybrids were capable of VCA expression, in contrast to the Raji parent that expressed EA but not VCA. A new Raji-Daudi hybrid (DITRUD-1) expressed spontaneous and induced EA and VCA at levels intermediate between its two parents. A different type of hybrid was derived from a Daudi subline and the K562 leukemia cell line (DUTKO-1) and was found to be capable of spontaneous as well as induced EA and VCA expression. Both DUTKO-1 and DITRUD-1 were similar to Daudi in their induction profile in respect to a very low response to TPA. PMID- 6278189 TI - Calorie restriction: effect on growth of human tumors heterotransplanted in nude mice. AB - The presence of 4 human malignant tumors (1 breast, 1 lung, and 2 colon carcinomas) growing subcutaneously as heterotransplants in nude mice did not significantly affect the body weights of adult animals until the tumors reached very large dimensions (tumor wt greater than 15% of the body wt). However, a colon carcinoma (HT 29) induced a cessation of the natural rate of body weight increase when it grew in young adults (animals weighing approximately equal to 25 g which will gain 6 g or approximately equal to 25% body wt in 1 mo). Calorie restriction at all the levels tested (8, 6, 4, and 2 g/day/mouse) with standard pelletized mouse food produced both weight loss in the animals (with and without tumor) and a lowering of the growth rate of all the 4 tumors tested growing at a subcutaneous site and/or under the kidney capsule. Each tumor responded differently to the calorie restriction. The 4 tumors tested grew equally in both male and female nude mice. Young animals weighing 20 g inoculated with a fifth tumor (MeWo melanoma) exhibited tumor growth inhibition proportional to restriction of calorie intake. Their survival, however, did not improve. PMID- 6278190 TI - Cholangiocellular carcinomas induced in Syrian golden hamsters administered aflatoxin B1 in large doses. AB - The hepatocarcinogenicity of aflatoxin B1 (AFB1) was compared in male Syrian golden hamsters and inbred F344 rats. AFB1 was administered by gavage 5 days/week for 6 weeks at doses of 1 and 2 mg/kg body weight/day to rats and hamsters, respectively; rate did not survive beyond 3 weeks with doses of 2 mg/kg/day. After 6 weeks the animals either received no further treatment or were given 0.1% phenobarbital sodium in the drinking water. ATPase-deficient foci of hepatic parenchymal cells, neoplastic nodules, and hepatocellular carcinomas were observed in liver sections from AFB1-treated rats killed at 6, 14, or 23 weeks; they were not seen in sections from AFB1-treated hamsters killed at 6, 14, or 46 weeks. Each of the 50 AFB1-treated rats developed hepatocellular carcinomas by 46 weeks; many also had cholangiocarcinomas and mixed hepatocellular cholangiocellular carcinomas. Hepatocellular carcinomas were found in only 2 of 49 AFB1-treated hamsters by 78 weeks. At this time cholangiocarcinomas were found in 15 hamsters, and microscopic cholangiomas were seen in all of the livers. Compared to the rat, the hamster was resistant to the hepatotoxic and hepatocellular carcinogenic effects of AFB1. PMID- 6278191 TI - Effects of dietary lipids on lactogenic hormone receptor binding in rat mammary tumors. AB - Both epidemiologic studies in humans and experiments in laboratory animals have indicated that high-fat (HF) diets promote mammary tumor growth; however, the biochemical mechanisms responsible for this accelerated tumor growth are poorly understood; thus this study was designed to determine whether diet-induced alterations in the lipid composition of mammary tumor cell membranes were associated with differences in lactogenic hormone binding capacity. Mammary tumors were induced with N-methyl-N-nitrosourea in 50-day-old female inbred Buffalo rats that were maintained on either HF or low-fat (LF) diets composed of either 20% corn oil or 0.5% corn oil, respectively. The microsome-membrane fractions of these tumors were then analyzed for specific lactogenic hormone binding with the use of 125-I-labeled human growth hormone. Methylated extracts of these same membrane fractions were also subjected to gas-liquid chromatography. Our results demonstrated that the mammary tumor membranes of the HF group did have a significantly greater lactogenic binding capacity than those of the LF group and that these differences in hormone binding were accompanied by significant alterations in the membrane qualitative fatty acid profiles of each group. Therefore, one way in which dietary lipids may be able to influence mammary tumor growth is by modification of the lactogenic hormone binding capacity of tumor cell membranes. PMID- 6278192 TI - Prolactin receptors in human breast tumors. AB - Prolactin-binding sites were determined in 759 human breast tumor biopsy specimens. Ovine prolactin (oPRL) was used in the binding assay of 569 tumors with 13% of the tumors showing a positive content with more than 1% specific binding. Human prolactin (hPRL) was utilized for 343 tumors with 36% of the tumors binding 1% or greater of the added radioactivity. When human growth hormone (hGH) was used as the labeled ligand in 95 tumors, only 2% of the tumors specifically bound more than 1% of the hGH. A total of 153 tumors were assayed simultaneously with the use of labeled hPRL and oPRL. Whereas 10.4% of the tumors showed the presence of prolactin-binding sites (greater than 1% specific binding) when oPRL was used, an approximately threefold increase (29.4%) in tumors possessing prolactin receptors was revealed when hPRL was used as the labeled ligand. Therefore, hPRL is the preferred ligand for the assessment of prolactin receptor levels in human breast cancer biopsy specimens. PMID- 6278193 TI - Tumor promoter 12-O-tetradecanoylphorbol 13-acetate: effect on complement and Epstein-Barr virus receptors in human lymphoblastoid cell lines. AB - The effect of the tumor promoter 12-O-tetradecanoylphorbol 13-acetate (TPA) on the C3 and Epstein-Barr virus (EBV) receptors in various human lymphoblastoid cells was investigated with the use of the erythrocyte-antibody-complement (EAC) rosette formation method and a quantitative bioassay for EBV receptors. TPA caused a significant decrease of C3 receptors in the cultures of both Raji and SB4 cells (by approximately 50% of the C3 receptors in untreated cultures at 10 ng/ml). Kinetic studies revealed that the rate of reduction was rather moderate but progressive, reaching a maximum 5 days after treatment with TPA. Kinetic studies showed that EBV receptors also decreased, similar to the reduction seen with C3 receptors after TPA treatment. The effect of TPA on the reduction of C3 receptors was observed not only in these cells but also in other EBV-positive B cells, subclones of SB4 cells, and MOLT-4 cells. However, in an EBV-negative B cell line, BJAB, EAC rosette formation was significantly enhanced. PMID- 6278194 TI - Epidemiological features of Wilms' tumor: results of the National Wilms' Tumor Study. AB - Nearly 2,000 children with Wilm's tumor registered in a national clinical trial during 1969-81 showed high rates of aniridia, hemihypertrophy, cryptorchidism, hypospadias, and other genitourinary anomalies. Patients with bilateral disease, who constituted 5% of the total, had younger ages at diagnosis and an increased incidence of congenital anomalies and renal blastemal rests. Those with multicentric unilateral lesions had more blastemal rests but were otherwise indistinguishable from the unicentric cases. The 20 familial cases had none of the features usually associated with genetic tumors: neither younger ages nor an increase in bilaterality nor associated congenital anomalies. These observations suggest that the fraction of Wilm's tumors that is due to an inherited mutation may be substantially smaller than previously supposed and support the concept that the disease arises from a variety of pathogenetic pathways. PMID- 6278195 TI - Glutathione S-transferase activity: enhancement by compounds inhibiting chemical carcinogenesis and by dietary constituents. AB - Benzyl isothiocyanate, beta-naphthoflavone, coumarin, alpha-angelicalactone, disulfiram, indole-3-carbinol and indole-3-acetonitrile induced increased glutathione (GSH) S-transferase activity in the liver and small intestine in female ICR/Ha mice. All seven compounds are inhibitors of chemical carcinogenesis. In additional work, several dietary constituents increased GSH S transferase activity. Consumption of diets containing dried powdered preparations of brussels sprouts, cabbage, coffee beans, or tea leaves resulted in increased GSH S-transferase activity. Mice fed an unrefined diet (Purina Rat Chow) had a higher GSH S-transferase activity than those fed a semipurified diet. The results of the present study indicated that the composition of the diet can alter the activity of an important enzyme system having the capacity to detoxify chemical carcinogens. PMID- 6278196 TI - Conditions associated with relapse of amphotericin B-treated disseminated histoplasmosis. AB - Progressive disseminated histoplasmosis (PDH) is a rare consequence of infection with Histoplasmia capsulatum. Usually fatal if untreated, PDH generally is cured by appropriate amphotericin B treatment. Of 31 persons with uncomplicated PDH treated with amphotericin B, we found that relapse occurred in five (16%) after an interval of up to nine years after initial therapy. Review of these five cases and 31 additional relapsing cases from the literature indicates that fungal endocarditis or endarteritis without surgical treatment, underlying lymphoreticular neoplasm, and amphotericin B dosage of less than 2 g appear to be associated with relapse of PDH. PMID- 6278197 TI - [Objective test of the effectiveness of surgical treatment in suppurative soft tissue processes of the face and neck]. PMID- 6278198 TI - [Encephalitis following measles-mumps vaccination simultaneous to an EBV infection (author's transl)]. AB - This is a case-report of a 15-month-old child, who received a measles-mumps vaccination in the course of an asymptomatic EBV-infection. Nine days later the infant incurred encephalitis and recovered substantially within three weeks; the EEG also returned to normal in this time. Ataxia still persisted, however, a half year later. The child produced serum antibodies against all three viruses, whereby the present low EBNA-titer is indicative of a chronic persistent EBV infection. This is, according to our knowledge, the first description of a EBV infection during a measles-mumps vaccination. PMID- 6278199 TI - [Nursing care of tumor patients]. PMID- 6278200 TI - [Game possibilities in nursing education]. PMID- 6278201 TI - [Functional scope of nurse anesthetists in Switzerland]. PMID- 6278202 TI - [Continued education of nurse anesthetists. Viewpoint of the Committee for the Education of Nurse Anesthetists]. PMID- 6278203 TI - [The concerns of nursing assistants]. PMID- 6278204 TI - [Viewpoint apropos of the article: "The concerns of nursing assistants"]. PMID- 6278205 TI - [The birth of CASSI (Collective of Promoters of Sessions in Nursing Care). The nurses take account]. PMID- 6278206 TI - [Enteral tube feeding]. PMID- 6278207 TI - [Towards a possible team care]. PMID- 6278208 TI - [The critical hour, between 10 and 11...]. PMID- 6278209 TI - [Research on the school-probation relationship]. PMID- 6278210 TI - [A theory of nursing care at the hospital]. PMID- 6278211 TI - [The 2 students delegated by the Swiss Nurses' Association write. ENSG-Meeting (European Nursing Student group) 1-8 October 1981 in Dublin, Ireland]. PMID- 6278212 TI - [Ideology of drug therapy]. PMID- 6278213 TI - [Effective decubitus prevention with supersoft mattresses and 30-degree slope position]. PMID- 6278214 TI - [St. Galler Nursing School students turn to the administrative adviser. Inconvenience allowance for nursing students]. PMID- 6278215 TI - [Role of the voluntary worker in community and hospital. The oldest form of nursing care]. PMID- 6278216 TI - [Theme-centered interaction according to R. C. Cohn and its application in nursing]. PMID- 6278217 TI - [Professional re-entry]. PMID- 6278218 TI - [A proposal for nursing education]. PMID- 6278219 TI - Viral particles in a papilloma from a green lizard (Lacerta viridis). PMID- 6278220 TI - Role of estrogens as promoters of hepatic neoplasia. AB - The administration of estrogens to humans has been associated with the development of benign and possibly malignant hepatocellular neoplasms. This study was designed to investigate the mechanism of this association. We present a rat model that demonstrates that stilbestrol (DES) and ethinyl estradiol promote the development of hepatic neoplasms in rats previously initiated with diethylnitrosamine (DEN). Eighty male and 12 female Fischer-344 rats were given a single intraperitoneal injection of either DEN (200 mg. per kg. of body weight) or saline. Beginning 4 weeks after injection, the rats were given an estrogen or corn oil twice weekly for up to 50 weeks. Treatments were stopped at the time of sacrifice or 11 to 29 weeks prior to sacrifice. Estrogen treatments included high dose DES (5 mg. per dose), low dose DES (0.5 mg. per dose), and ethinyl estradiol (0.2 mg. per dose). Male and female rats given both DEN and high dose DES developed grossly visible hepatic hyperplastic nodules (mean, 9.1 per liver) after 20 weeks of DES. Nodules also developed if the onset of DES treatment was delayed until 28 weeks after initiation. Rats given DEN alone or DES alone did not develop nodules after 20 weeks. Microscopic hyperplastic foci also developed in rats given DEN plus DES, DES alone, and DEN alone. The foci in rats given DES alone were largely reversible on cessation of estrogen therapy. Mitotic activity in foci and nodules was prominent during estrogen therapy but declined markedly after cessation of therapy. Similar promoting activity of ethinyl estradiol was observed. Low dose DES was not effective in promoting tumor formation. The data indicate that estrogens promote hepatic neoplasia, perhaps by increasing hepatocyte mitotic activity and thereby facilitating the evolution of previously initiated cells into neoplastic clones. Comparison with human disease should be made with caution, especially because the estrogenic dose administered was approximately 200-fold the usual contraceptive dose in humans. However, the analogy between this model and the development of human oral contraceptive associated hepatic tumors is evident. It is possible that some women have latent "initiated" cells that divide faster than the surrounding hepatocytes in response to estrogens. PMID- 6278221 TI - A survey of leprosy deformities in a closed community. AB - Out of 1011 leprosy patients, 200 (20%) cases with permanent deformities were found in the Leprosy Control Unit, Raxaul (East Champaran), Bihar. The statistical data of selected patients and the types of various deformities are presented. The prevalence rate and incidence pattern of deformities reported by other workers are compared. PMID- 6278222 TI - In vivo and in vitro metabolism of estrone and estradiol-17 beta and their 3 sulfates in pregnant female guinea pigs: a plausible prehormone role of estrogen sulfates in the maternal uterus. PMID- 6278224 TI - Receptor for 1,25-dihydroxyvitamin D3 in GH3 pituitary cells. AB - Cytosol prepared in 0.3 M KCl from pituitary GH3 cells, but not from AtT-20 cells contains a receptor-like macromolecule that binds 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) with specificity and high affinity (Kd = 2.9 x 10(-10) M). The GH3 cytosolic binding component sediments at 3.3 S in high-salt sucrose gradients and adsorbs to DNA-cellulose; its elution profile from DNA-cellulose and other biochemical properties are indistinguishable from those of classical 1,25(OH)2D3 hormone receptors. The presence of the 1,25(OH)2D3 receptor in pituitary cells which secrete primarily growth hormone and prolactin (GH3), but not in a line which secretes the 31,000-dalton ACTH precursor and its derived peptides (AtT 20), suggests that 1,25(OH)2D3 may play a regulatory role in specific pituitary cells. PMID- 6278223 TI - Characterization of rat capsular adrenal (zona glomerulosa) (Na,K)-ATPase activity. AB - A higher (Na,K)-ATPase activity was found in rat capsular (zona glomerulosa) than in decapsulated (zona fasciculata) adrenal. No unusual characteristics of this enzyme were found in the simulation with Na+ and K+ and the inhibition with ouabain. Angiotensin II, ACTH and serotonin, all potent stimulators of aldosterone biosynthesis, did not affect the enzyme. In conclusion, the characteristics of rat capsular adrenal (Na,K)-ATPase are identical to the classical enzyme. It is not the direct receptor or effector for stimulators of aldosterone biosynthesis but a supportive role in mediating the regulatory signal cannot be ruled out. PMID- 6278225 TI - The enzyme specificity of ACTH stimulation of rabbit adrenal microsomal 17 alpha hydroxylase activity. PMID- 6278226 TI - Prediction of outcome in treatment of alcoholism: a study of staff and patients. PMID- 6278227 TI - Time-dependent extracellular ion transport. PMID- 6278228 TI - Catastrophe theory of dopaminergic transmission: a revised dopamine hypothesis of schizophrenia. PMID- 6278229 TI - A paucity of palindromes in phi X174. PMID- 6278230 TI - Lung cancer in young persons. AB - Lung cancer in patients under 40 years old is rare. Among 718 patients with lung cancer, 5% or 35 patients were younger than 40. All but one were heavy smokers. Twelve young patients who had operations survived 41.7 (SD 46.3) months: they included six who had resections of Stage 3 disease and who survived 19.2 (SD 16.0) months. The other six young patients treated operatively were free of nodal metastasis N0); all survived more than 3 years and three of them are apparently cured. The 23 young patients who were not treated operatively survived for 5.6 (SD 3.1) months. The 5 year survival rate of these young operated patients was not different from that of 201 operated patients over 40 years of age. Young nonoperated patients survived for a significantly shorter time (p less than 0.0001) than did the older patients who also received only chemotherapy and/or radiation as a treatment. In young patients adenocarcinomas predominated (48.6%), and the incidence of small cell undifferentiated cancers was high (28.6%). These significant differences as compared to the control group did not explain the short survival time of the young patients treated nonsurgically. We conclude that lung cancer in young persons is virulent and that diagnosis is frequently delayed. Therapy, in selected patients, should include aggressive resection, sometimes despite advanced local disease. This group of patients justifies innovative, intensive efforts at more prompt diagnosis and experimental multimodal therapy. PMID- 6278231 TI - Bronchogenic carcinoma located in the aortic window. The importance of the primary lesion as a determinant of survival. AB - Thirty-four patients with an aortic window lesion were carefully staged with gallium scans and mediastinoscopy according to the TNM classification system for carcinoma of the lung. All were in Stage III. Twenty-five patients had non-oat cell carcinomas (15 squamous cell, eight adeno-, two large cell) and nine had oat cell carcinomas. Quantitative ventilation-perfusion lung scans were particularly helpful in verifying the subaortic location of the tumor by showing a less than 20% interference with pulmonary blood flow or ventilation secondary to left mainstem bronchus or pulmonary artery invasion. Decision for resectability in 13 Stage III M0 patients was based on the length of the uninvolved proximal left main pulmonary artery seen on pulmonary arteriogram. Eight patients (seven non oat cell and one oat cell) had resection after radiation and prior to chemotherapy (after two cycles of chemotherapy and prior to radiation therapy for the oat cell) with a resultant survival rate better than those of M0 and M1 non oat cell or oat cell patients without resection. The survival rates of nine non oat cell M0 patients, nine non-oat cell M1 patients, and eight oat cell patients, all without resection, were not statistically different. This similarity in survival rates is explained by the observation that 38% of the non-oat cell M1, 71% of the non-oat cell M0, and 63% of the oat cell patients died from complications of their primary tumor. Patients with aortic window lesions, irrespective of their histology, have an extremely poor prognosis due to the high incidence of lethal complications of their primary tumor. Complete resection when feasible, as judged by pulmonary arteriography, provides the best control of the primary tumor and, as a consequence, gives longer survival. PMID- 6278232 TI - Photoreactivation in two closely related marine fishes having different longevities. AB - The tautog, Tautoga onitis, and cunner, Tautogolabrus adspersus, are marine fishes which are closely related taxonomically (family Labridae), have similar habits, the same ecological requirements and are found in the same areas. However, they differ markedly in their longevity. The tautog is longer-lived, reaching an estimated average age of 34 years. It attains sexual maturity at 3--4 years of age. The cunner, on the other hand, lives to about 5--6 years of age and is sexually mature at about one year. An examination of their respective capacities for photoreactivation using cells in tissue culture derived from these fishes revealed marked differences in the rate of monomerization of ultraviolet induced pyrimidine dimers from their DNA. Radiochromatographic analysis of dimers in the tautog showed that half the dimers were removed within a period of 10 min of exposure to photoreactivating light whereas a 50-min exposure was required to photoreactivate dimers to this same level in the cunner. Dark repair of dimers was measured by radiochromatography and by endonuclease assay. The two methods showed similar low levels of excision repair in both species. PMID- 6278233 TI - A study of the mechanism of estrogen as an antiatherosclerotic: the inhibitory effect of estrogen on A23187-induced contraction of the aortic wall. AB - Although the antiatherosclerotic effects of estrogen are well known, the mechanisms involved have not been determined. We designed experiments in which strips of thoracic aorta from oophorectomized rabbits were contracted by the calcium ionophore A23187 and the inhibition of the contraction by estrogen and concomitant levels of cyclic nucleotides were determined. A23187 (10(-5) M) increased the tension slowly but progressively, and there was a concomitant decrease in cyclic AMP levels. While a dose-dependent relaxation occurred in strips contracted by A23187 with addition of theophylline (greater than 10(-4) M) or dibutyryl cyclic AMP (greater than 10(-4) M) to the bath, prior addition of these compounds inhibited the contraction. Pretreatment of the strips with estrogen (2 x 1-(-7) M, 2 x 10(-5) M) significantly prevented contraction of the strips, as induced by A23187, and also prevented reduction in cyclic AMP levels, as induced by the ionophore. Cyclic GMP levels remained unchanged throughout. As the contraction of these tissues wall was inhibited by estrogen in an apparent association with a reduction in the levels of cyclic AMP, this might help to explain the antiatherosclerotic effects of estrogen, as reduction in these nucleotide levels has been suggested to induce atherosclerotic lesions in the arterial wall by enhancing vascular permeability. PMID- 6278234 TI - Modulation of age-related loss of glucagon-promoted lipolysis by food restriction. AB - Male Fischer 344 rats were fed ad libitum (Group A) or 60% of the ad libitum intake (Group R) starting at 6 weeks of age. The latter is a dietary manipulation that prolongs life. At 6, 10 and 15 weeks of age, rats were killed, free adipocytes prepared, and lipolytic, adenylate cyclase, and phosphodiesterase activities of adipocytes measured. The ability of glucagon to promote lipolysis markedly declined between 6 and 15 weeks of age in adipocytes from Group A rats but not in those from Group R rats. Glucagon-promoted adenylate cyclase activity paralleled lipolytic activity. Phosphosdiesterase activities increased with age, to a greater extent in adipocytes from Group A rats than in those from Group R rats. It is concluded that the loss in glucagon-promoted lipolysis with age in adipocytes from Group A rats is primarily the result of either a loss of or a change in the characteristics of the glucagon receptors or an alteration in the system coupling the receptors with adenylate cyclase and that food restriction prevents these changes. The relevance of these findings in analysis of the mode by which food restriction delays the aging process is discussed. PMID- 6278235 TI - [The effect of hypoxia on peripheral motor nerve (n. ulnaris) latencies in alcoholic polyneuropathies]. PMID- 6278236 TI - Polymyxin B, colistin, and sodium colistimethate. PMID- 6278237 TI - Effect of disulfiram and its reduced metabolite, diethyl-dithiocarbamate on aldehyde dehydrogenase of human erythrocytes. AB - Inhibition of human erythrocyte aldehyde dehydrogenase (ALDH) activity was studied using disulfiram and its reduced metabolite, diethyldithiocarbamate (DDC). The enzyme was rapidly inactivated by disulfiram and the inhibition was protected by reduced glutathione (GSH), in a concentration dependent manner when the enzyme premixed with GSH was reacted with disulfiram. Higher reactivity of the thiol group of the enzyme than that of GSH to disulfiram was suggested from the observation that half of the enzyme activity was inhibited when the ratio of disulfiram to GSH was 1:10. Although DDC alone showed no inhibitory effect on the enzyme, inactivation was mediated by a low concentration of heme-containing peroxidases, but not by methemoglobin. Under this condition, the inhibition potential was not protected, even with a high concentration of GSH. The constant reoxidation system of DDC is probably directly related to the enzyme inactivation. PMID- 6278238 TI - Human platelet vasopressin receptors. AB - Specific saturable binding of 125I-arginine-vasopressin to human platelets is described. For ten normal volunteers the mean (+/- S.D.) KD is 5.6 nM (+/- 2.1) and the mean (+/- S.D.) Bmax is 115 fmoles/mg protein (+/- 30). Association studies indicate that equilibrium is reached after 90 minutes on ice. Pharmacological inhibition studies with analogues indicate that the platelet receptor is very similar to the kidney medulla receptor. The function of the receptor may involve serotonin release and platelet aggregation. Vasopressin binding to platelets should provide a readily means of assessing vasopressin receptor function in man. PMID- 6278239 TI - Bacterial haem-iron detection using occult blood tests. PMID- 6278240 TI - Fire deaths in the Glasgow area: III. The role of hydrogen cyanide. PMID- 6278241 TI - Physical mapping of the srl recA region of Escherichia coli: analysis of Tn10 generated insertions and deletions. AB - A restriction endonuclease map for the enzymes EcoRI, BamHI, SalI, and PstI covering 23.5 kilobase pairs (kb) of the srl recA region of Escherichia coli was constructed. An insertion of the transposon Tn10 in the negative regulatory gene srlR was shown to be located 5.8 kb away from the promoter proximal end of the recA gene. The extent of several Tn10 generated deletions, originating from the srlR301::Tn10 insertion, were analyzed by physical mapping. Three mutations that had removed the Tn10 encoded tetracycline resistance gene, del(srl-recA)302, del(srl-recA)304, and del(srl-recA)303, were found to be deleted for 40%, 45%, and 50% of the recA structural gene, respectively. A deletion, del(srl-recA)306, that had not affected the structure of the Tn10 in srlR301 was shown to have removed the entire recA structural gene. PMID- 6278243 TI - The use of cloned nif (nitrogen fixation) DNA to investigate transcriptional regulation of nif expression in Klebsiella pneumoniae. AB - Some restriction endonuclease fragments of nif DNA, when carried on small multicopy plasmids, inhibited nif expression in Klebsiella pneumoniae. A study of this inhibitory effect revealed, (1) that overproduction of the nifL gene product inhibited transcription of two nif operons examined, nifJ and nifHDKY and, (2) that when transcription was initiated from the promoter of the nifHDKY operon on multicopy plasmids there was a corresponding decrease in the transcription rates of the chromosomally located nifJ and nifHDKY but not the nifLA operon. Studies of transcription in vivo also showed that the nifA gene product was essential for transcription initiation from the nifHDKY and nifBQ promoters. These results, taken with earlier observations (see Discussion) provide evidence that the nifL and nifA gene products are respectively a repressor and activator of nif transcription initiation from all nif promoters except that of the nifLA operon. PMID- 6278242 TI - Cointegrates between bacteriophage P1 DNA and plasmid pBR322 derivatives suggest molecular mechanisms for P1-mediated transduction of small plasmids. AB - We characterized cointegrates formed in an Escherichia coli rec+ strain between bacteriophage P1 genomes and small plasmids related to pBR322. The partners were, on the one hand, either phage P1 DNA, which carries one copy of IS1, or phage P1 15 DNA, a derivative which lacks the IS1, and, on the other hand, plasmids containing either a split IS1 or no. In the presence of IS1 sequences on both partners, cointegrates were usually formed by reciprocal recombination between SI1 sequences. Cointegrates between P1 and a plasmid carrying no IS1 sequence were formed by transpositional cointegration mediated by IS1 of P1. Cointegrates between P1-15 and small plasmid containing a split IS1 were formed by one of three ways: (a) acquisition of an IS1 by P1-15 followed by reciprocal recombination between IS1 sequences, (b) transpositional cointegration mediated by the split IS1 element, Tn2657, or (c) involvement of the invertible segment carried on P1-15 DNA. Most cointegrates segregated into the small plasmids and phage P1 derivatives. A comparison of the phenomenon studied and of their frequencies allowed us to conclude that cointegrate formation is a molecular mechanism involved in the transduction of plasmids smaller than those packageable into P1 virions, although it does not seem to be the only process used. PMID- 6278244 TI - Physical characterization of the plasmid pON5300. AB - The antibiotic resistance plasmid R1drd19Km- which has spontaneously lost its kanamycin resistance marker, and its derivative pON5300, were analysed using the restriction endonucleases SalI, BamHI, HindIII and EcoRI. The fragment patterns were compared with that of the R1drd19 and the fragments responsible for kanamycin resistance were found to be missing in R1drd19Km- and pON5300. In these plasmids a 7 Mg/mol EcoRI fragment was observed instead of the D (6.3 Mg/mol) fragment of R1drd19. Further a new 6 Mg/mol EcoRI restriction fragment was observed in pON5300. Using double digestions it was shown that the new fragment does not carry restriction sites for HindIII, BamHI and SalI endonucleases. The non-homology of the analysed plasmid was proved electron microscopically by heteroduplex techniques. The possibility of amplification in the regulatory region for the expression of R-determinants in pON5300 is discussed. PMID- 6278245 TI - Location of antibiotic resistance determinants, copy control, and replication functions on the double-selective streptococcal cloning vector pGB301. AB - The physical map of the streptococcal cloning vector pGB301 (9.8 kb) has been extended by localizing the cleavage sites of restriction endonucleases AvaII, AvaI, BclI, BstEII, and PvuII. The latter four enzymes cleaved pGB301 at unique sites. Insertion of chromosomal DNA from Staphylococcus aureus strain 3Ar-m- into the single BstEII site of pGB301 led to inactivation of the plasmid's chloramphenicol resistance determinant. Twelve deletion derivatives of pGB301 were isolated either by in vitro manipulation of pGB301 or as spontaneous deletion mutants following transformation of Challis by mixtures of recombinant plasmids. The overlapping deletions which spanned a continuous sequence of 7.7 kb ranged in size from 0.3 kb to 4.1 kb and allowed to localize the chloramphenicol and MLS-resistance determinants, the copy control function, and the replication region on the physical map of pGB301. Plasmid pGB301 together with its deletion mutants constitutes a valuable tool for further molecular cloning experiments in streptococci. PMID- 6278246 TI - Maintenance and genetic stability of vector plasmids pBR322 and pBR325 in Escherichia coli K12 strains grown in a chemostat. AB - The maintenance and genetic stability of the vector plasmids pBR322 and pBR325 in two genetically different Escherichia coli hosts were studied during chemostat cultivation with glucose and ammonium chloride limitation and at two different dilution rates. The plasmid pBR322 was stably maintained under all growth conditions tested. However pBR325 segregated from both hosts preferentially during glucose limitation and at low dilution rate. In addition to this general segregation process a separate loss of tetracycline resistance was observed. The remaining plasmid conferred resistance to ampicillin and chloramphenicol only, without any remarkable alteration of its molecular weight. Cultivation conditions in the chemostat were found that allowed the stable genetic inheritance of both plasmids in the hosts studied. PMID- 6278247 TI - The nucleotide sequence of the atp genes coding for the F0 subunits a, b, c and the F1 subunit delta of the membrane bound ATP synthase of Escherichia coli. AB - The nucleotide sequence has been determined of a 2,500 base pair segment of the E. coli chromosome located between 3.75 and 6.25 kb counterclockwise of the origin of replication at 83.5 min. The sequence contains the atp genes coding for subunits a-, b-, c-, delta- and part of the alpha-subunit of the membrane bound ATP synthase. The precise start positions of the atpE (c), atpF (b), atpH (delta) and atpA (alpha) genes have been defined by comparison of the potential coding sequences with the known amino acid sequence of the c-subunit and the determined N-terminal amino acid sequences of the respective subunits. The genes are expressed in the counterclockwise direction. Their order (counterclockwise) is: atpB (a), atpE (c), atpF (b), atpH (delta) and atpA(alpha). The coding sequences for subunits b and delta yield polypeptides of 156 and 177 amino acids, respectively, in accordance with the established sizes of these subunits; the one for the c-subunit, the DCCD binding protein, fits perfectly with its known sequence of 79 amino acids. The a-subunit is comprised within a coding sequence yielding a polypeptide of 271 amino acids. It is suggested, however, that the a subunit (atpB) contains only 201 amino acids, in accordance with its known size, starting from a translation initiation site within the larger coding sequence. The stoichiometry of the F0 sector subunits is discussed and a model is proposed for the functioning of the highly charged b-subunit of the F0 sector as the actual proton conductor. PMID- 6278248 TI - Tn1 insertion mutagenesis in Escherichia coli K-12 using a temperature-sensitive mutant of plasmid RP4. AB - A method for Tn1 insertion mutagenesis in Escherichia coli has been developed using pTH10, a mutant plasmid of RP4 temperature-sensitive for maintenance. The mutagenesis involves three steps. Firstly, from strains carrying pTH10 showing resistance to the antibiotics kanamycin, tetracycline, and ampicillin at 30 degrees C but not at 42 degrees C, clones are isolated resistant to kanamycin at 42 degrees C. Such temperature-independent, drug resistant clones probably carry pTH10 integrated into the host chromosome. Secondly, they are cultivated at 30 degrees C. At this temperature segregants carrying pTH10, which has been excised from the host chromosome, accumulate. Thirdly, to cure such segregants of autonomous pTH10, they are cultivated at 42 degrees C. By these procedures, clones free of pTH10, but carrying Tn1 insertions on the host chromosome, were obtained. About 3% of the clones carrying Tn1 insertions were auxotrophic. Distribution of auxotrophic mutations was not random, indicating the existence of preferential integration sites of Tn1 on the host chromosome. The frequency of precise excision of Tn1 was less than 10(-10). The pTH10 plasmid has a wide host range among Gram-negative bacteria and thus may serve as a excellent vector for insertion mutagenesis of Tn1 in many Gram-negative bacterial species. PMID- 6278249 TI - The location of sequences of TnA required for the establishment of transposition immunity. PMID- 6278250 TI - The tnpR gene product of TnA is required for transposition immunity. AB - A mutant of TnA no longer recognizing immune plasmids had been isolated. The mutation is complemented in trans by a functional tnpR gene. The requirement for wild type tnpR gene product for the establishment of transposition immunity was confirmed by the use of a derivative of transposon Tn3 in which both the tnpA and the tnpR genes are partly deleted. This deleted Tn3 was shown to transpose onto an immune plasmid in the presence of a wild type tnpA gene but not in the presence of both tnpA and tnpR genes. PMID- 6278251 TI - A fine structure map of spontaneous and induced mutations in the lambda repressor gene, including insertions of IS elements. AB - Mutations at over 70 sites in the cI gene have been mapped by 4-factor crosses and assigned precise or approximate positions in the DNA sequence. 16 of 25 spontaneous mutations were insertions of IS1, IS3 or IS5 into AT-rich regions of cI. The 5-methylcytosine in the sequence Cm5CAGG is a hot spot for spontaneous cI amber mutations. Recombination frequencies between mutations were proportional to distance with the exception of amber mutations at 4 sites, including the host spot for spontaneous mutations. Mutations with a given phenotype are clustered on the genetic map. No missense mutations affecting repressor activity were found in the central one-third of cI, but 5 of 6 ind- mutations were located in this region. The amino-terminal third of the gene contains the sites of most trans dominant cI- mutations, and of all ts mutations that result in repressors that are reversibly inactivated at high temperatures. PMID- 6278253 TI - Cloning of a eukaryotic regulatory gene. AB - From a pool of hybrid plasmids carrying Sau3A fragments representing the entire yeast (S. cerevisiae) genome, a DNA fragment containing the regulatory gene PPRI was cloned by complementation of a non-inducible ppr1 mutation which confers to the cells an increased sensitivity to 6-azauracil. Cells containing the cloned DNA regained the ability to induce the synthesis of URA1 and URA3 gene products controlled by PPR1. A physical map has been constructed and the study of subcloned restriction endonuclease fragments from the original yeast DNA fragment allowed us to localize the wile-type PPR1 regulatory gene within a 3 kilobase pair region. The ppr1 RNA level was measured and the hybridization data indicate in a wild-type strain a low efficiency of transcription of PPR1 as compared to the structural URA3 gene, without effect of inducing conditions. PMID- 6278252 TI - Expression of the Herpes simplex virus thymidine kinase gene in Saccharomyces cerevisiae. AB - Yeast plasmids have been constructed that carry the Herpes simplex Virus type 1 (HSV-1) thymidine kinase (TK) gene which is functionally expressed in Saccharomyces cerevisiae. The expression of the TK gene appears to be due to transcriptional read-through from a yeast promoter that lies on the 3' side of the HIS3 gene. The TK+ yeast possesses in vitro thymidine kinase activity which is absent in the original yeast strain. Yeast strains auxotrophic for thymidine monophosphate (dTMP) (tmp1) can grown on thymidine-containing medium after transformation with these plasmids. Tmp+, TK+ S. cerevisiae whose de novo synthesis of dTMP is inhibited with amethopterin plus sufanilamide is also capable of growth in thymidine. S. cerevisiae transformed with such plasmids is capable of incorporating thymidine and bromodeoxyuridine into DNA. PMID- 6278254 TI - Plasmid transformation in Bacillus subtilis. The significance of partial homology between plasmid and recipient cell DNAs. AB - A series of hybrid plasmids consisting of pC194 or pUB112 and B. subtilis DNA were constructed. In contrast to plasmid pC194, purified monomeric forms of such plasmids were active in transformation, provided the recipient cells were recombination proficient. Similarly the monomers of pC194 derived plasmids, containing bacteriophage phi 105 DNA were able to transform phi 105 lysogenic but not nonlysogenic cells. From the results it is concluded that the presence of DNA/DNA homology between chromosomal DNA of the recipient cell and part of the hybrid plasmids used is a sufficient condition to endow monomeric plasmids with transforming activity. PMID- 6278255 TI - Plasmid transformation in Bacillus subtilis. Alterations introduced into the recipient-homologous DNA of hybrid plasmids can be corrected in transformation. AB - Various alterations (deletions, additions, inversions) were introduced into portions of pC194/B. subtilis or pC194/phi 105 hybrid plasmid molecules which are homologous to the DNA of recipients in transformation. These plasmids are stably maintained in transformations of recombination deficient cells. In transformations of recombination proficient cells, they can be corrected to those plasmid forms into which the alterations were originally introduced. This correction is most pronounced when transformations are performed with monomeric ccc forms of hybrid plasmids. It is suggested that correction is a consequence of mismatch repair occurring in the synapsis of homologous portions of plasmid and resident DNAs. PMID- 6278256 TI - Molecular cloning of the tolC locus of Escherichia coli K-12 with the use of transposon Tn10. AB - We have cloned the tolC gene of E. coli K-12 into pSF2124 by using transposon Tn10 as the marker to first isolate the relevant DNA fragment. The gene is on a 10.5 kb EcoRI fragment, and Tn5 insertion mutagenesis locates the gene near one end of this EcoRI fragment. An EcoRI-PstI fragment has been subcloned into pBR322 to facilitate further analysis of the gene. PMID- 6278257 TI - A system for measuring the frequencies of tandem and non-tandem double base substitutions induced by ultraviolet irradiation. PMID- 6278258 TI - Multiply branched replicative intermediates in E. coli and bacteriophage lambda. AB - Multiple branched DNA fragments present in a fast sedimenting complex comprising a minute fraction of the E. coli genome have been isolated. Similar structures were also observed among bacteriophage lambda DNA replicative intermediates after infection of synchronized E. coli cells. These structures were found to be associated with the amino acid and thymidine starvation steps required for synchronization and originate either by initiation from secondary sites or by snap-back of daughter strands containing substantial single stranded regions in the vicinity of the growing point. PMID- 6278259 TI - Cloning of mitochondrial DNA from the petite negative yeast Schizosaccharomyces pombe in the bacterial plasmid pBR322. AB - The entire mitochondrial (mt) genome of the yeast Schizosaccharomyces pombe (S. pombe) was cloned in the BamHI site of the Escherichia coli plasmid pBR322. Three lines of evidence demonstrate that the complete mtDNA molecule was amplified without rearrangement or partial loss. First, restriction of the hybrid plasmid with BamHI led to the recovery of two fragments corresponding to the linearized plasmid and the BamHI-cut mtDNA. Second, restriction of cloned and native mtDNA with HindIII revealed identical fragments. Third, mitochondrial ribosomal RNA hybridized to the same HindIII fragments from cloned mtDNA and from mtDNA isolated from mitochondria. PMID- 6278260 TI - rho Mutations restore lamB expression in E. coli K12 strains with an inactive malB region. AB - lamB, the structural gene for lambda receptor, is the second gene of the malK lamB operon in the malB region of the Escherichia coli K12 chromosome. lamB is essentially not expressed in the absence of an active malT gene product, the activator of the maltose regulon. A malT strain is resistant to phage lambda. We show that: (i) Introduction of rho mutations in malT mutants restores lamB expression to a level sufficient to render the strain sensitive to phage lambda; (ii) This restoration is not dependent on the main promoter of the malK lamB operon. It depends on the distal part of the malK gene. We propose that rho inactivation unmasks the activity of a promoter located near the distal end of malK. Experiments with Mu insertions in gene malK suggest that in the (-) orientation a Mu promoter is also able to allow lamB expression in a rho background. PMID- 6278261 TI - Mapping of mglB, the structural gene of the galactose-binding protein of Escherichia coli. AB - The tetracycline resistance transposon Tn10 was inserted into the E. coli chromosome near mglB550, a structural gene for the galactose-binding protein. P1 transductions established the position of these Tn10 insertions (zee-700, 701, 702::Tn10) close to the genes ptsF, fpk, cdd, mglB550, his, and gatA with 85% 95%, 85%, 36%, 20%-40%, 12%-15%, and 0.5% cotransduction frequency. Three factor crosses revealed the relative sequence of the genes as: mglB550, zee-700::Tn10, ptsF, fpk, cdd, his, gatA was found to be 1.3% cotransducible with mglB550. Two Tn10 insertions near gatA were isolated and characterized. One, zef-704::Tn10, was 3% cotransducible with fpk, 8% with mglB550, and 42% with gatA. The other, zef-703::Tn10, was 98% cotransducible with gatA but not with mglB550 or fpk. Neither of these two Tn10 insertions was cotransducible with cdd. Four factor crosses revealed the sequence gatA, zef-704::Tn10, mglB550, fpk. Neither zee 700::Tn10 nor zef-703::Tn10 showed an (0/300) cotransduction with either glpT or gyrA. The clockwise order of genes is then: his, cdd, fpk, ptsF, zee-700::Tn10, mglB550, zef-704::Tn10, gatA. With a fix-point for his at 44 min, fpk would be placed at 45 min and mglB550 at 45.5 min. During the course of this work we noticed that the cotransduction frequency between Tn10 insertions and nearby markers tended to increase when new P1 lysates were prepared from freshly reisolated strains. This may indicate loss of nonessential genes adjacent to Tn10 insertions. Using insertion zee-703::Tn10, we isolated deletions extending into an mgl gene other than mglB. Crosses between such a deletion mutant and an mglB550 mutant were done. The analysis of the periplasmic proteins of these as well as other transductants or recombinants involving the mglB550 or the mglB551 gene revealed the existence of strains synthesizing both the wild-type as well as the corresponding mutant protein. Strains containing both proteins exhibit either wild-type or mutant phenotype. These strains appeared unstable. Upon reisolation from purified stock cultures kept in glycerol at -20 degrees C, colonies could be isolated that carried only mutant or wild-type protein. PMID- 6278263 TI - Genetic instability at the cut locus of Drosophila melanogaster induced by the MR h12 chromosome. AB - Unstable mutations were generated at the cut locus by the MR-h12 factor which induces male recombination. The unstable allele ctMR2, containing the MR transposon in the cut locus is a very powerful mutator producing a number of different viable and lethal mutations both in the cut locus and outside it. I describe several types of mutations: stable reversion to wild type, which were sometimes associated with the appearance of unstable mutations in other loci; of stable deficiencies at the cut locus (lethals); new unstable mutations at different loci with the ctMR2 allele conserved; new unstable cut alleles with a phenotype other than that of ctMR2. The possible mechanisms of these mutational events are discussed. The genetic system constructed in the present work affords an opportunity for molecular studies of the cut locus and the MR-transposon, as a sequence from the cut locus has recently been cloned (Tchurikov et al. 1981). PMID- 6278262 TI - Primary structure of Escherichia coli RNA polymerase nucleotide substitution in the beta subunit gene of the rifampicin resistant rpoB255 mutant. AB - The transducing phage lambda dsupM814 and the plasmid pIB1830 containing the wild type rpoB gene have been constructed and the primary structure of the gene's central fragment has been established. In contrast with the wild-type, the gene of the rpoB255 mutant, whose primary structure has been published, was found to contain an A.T. leads to T.A. transversion entailing the substitution of a valine residue for the aspartic acid residue (516) of the wild-type beta subunit. PMID- 6278264 TI - Internal promoters of the rpoBC operon of Escherichia coli. AB - Four ribosomal protein genes, rplA, rplJ, rplK and rplL form an operon in E. coli together with the genes rpoB and rpoC which encode the beta and beta' subunits of RNA polymerase. Transcription is initiated principally at two promoters, PL11 and Pl, the overall structure of the operon being (in the direction of transcription) PL11 rplK rplA Pl rplJ rplL rpoB rpoC. Here we describe studies of phage lambda derivatives carrying various segments of this operon, which demonstrate the existence of at least three additional weak internal promoters, and help to define their positions and strengths. PMID- 6278265 TI - Nucleotide sequence of the 3'-terminal region of rat 18S ribosomal DNA. AB - The 3'-terminal 230 base-pairs (bp) of the gene for 18S rRNA and 40 bp of the adjoining spacer have been sequenced for the Sprague-Dawley rat. This mammalian sequence has been compared with the known sequences of yeast, fruit fly, silkworm, and frog. This study has shown that the nucleotide-sequence differences between rat and frog are the smallest among and longer species, probably reflecting their evolutionary closeness and longer maturation time compared to the others. There is little similarity in the nucleotide sequences of the transcribed spacer regions of the five species compared. PMID- 6278266 TI - Effects of various oestrogens on circulating androgens and cortisol during replacement therapy in post-menopausal women. AB - The influence of various oestrogens during unopposed replacement therapy on circulating androgens and cortisol was studied in 65 post-menopausal women. As dose dependent decrease in dehydroepiandrosterone sulphate (DHAS) was found. Ethinyloestradiol (0.05 mg daily) already gave a significant decrease after 1 mth of treatment. The decline following 17 beta-oestradiol (2 mg) and oestrone sulphate (2.5 mg) was less pronounced. Oestriol (6 mg daily) had no effect. Ethinyloestradiol also increased the levels of total cortisol and testosterone, probable because of serum protein induction, while 17 beta-oestradiol had no significant effect. Serum levels of androstenedione remained unchanged during therapy. PMID- 6278268 TI - The puzzle of pain. A continuing enigma. PMID- 6278267 TI - Conservative treatment of urinary incontinence in women with special reference to the use of oestrogens. AB - Urinary incontinence was treated conservatively in 100 patients. The follow-up period was 12-24 mth. For post-menopausal women, the oestrogen therapy consisted of oral oestradiol valerate or vaginal oestrone sulphate combined with emepronium bromide. In post-menopausal patients the best results were noted when incontinence had begun at the menopause and when the duration of the complaint was not more than 3 yr. For pre-menopausal patients, the treatment given was generally emepronium bromide. During the follow-up period 15 of the patients, 11 of whom were post-menopausal, became symptomless and 77 improved; that is, the incontinence was only slight and occasional. The treatment was without any effect in 8 of the patients. Oestrogen therapy was successful in most post-menopausal women. In these patients, the best results were obtained when the duration of the incontinence was not more than 3 yr. PMID- 6278269 TI - Endorphins and anesthesia. PMID- 6278270 TI - Expression of herpes simplex virus type-common surface antigens in clonal cells of an HSV type 2-transformed line: effect of adriamycin. AB - The expression of herpes simplex virus (HSV) type-common surface antigens (CSA) in a representative cell clone (155-4-03) of hamster cell line 155-4 transformed by HSV type 2 was enhanced by treatment with inhibitors of RNA synthesis [adriamycin (ADM) and daunomycin] but not with inhibitors of DNA synthesis (2 iododeoxyuridine, bleomycin, mitomycin C and cytosine arabinoside), although all these drugs decreased the number of viable cells to a similar extent. ADM enhanced CSA expression in the clone was inhibited by puromycin and 2-deoxy-d glucose, suggesting that the enhanced expression required both protein synthesis and glycosylation. This enhanced expression was sensitive to protease inhibitors (antipain and p-nitrophenyl-p'-guanidinobenzoate) and procaine, which is known to inhibit trypsin action and the organization of cell membrane-associated cytoskeletal elements (microfilaments and microtubules). Furthermore, low concentrations of ADM (0.1 microgram/ml) and actinomycin D (0.5 microgram/ml) enhanced CSA expression additively, but the most effective concentrations of ADM (0.25 microgram/ml) and actinomycin D (2 microgram/ml) did not. These findings indicated that the two drugs enhance CSA expression in the clone by a common mechanism. PMID- 6278271 TI - Grouping of adenoviruses and identification of restriction endonuclease cleavage patterns of adenovirus DNAs using infected cell DNA: simple and practical methods. AB - Simple and practical methods for grouping of adenoviruses and for identification of restriction endonuclease cleavage patterns of viral DNA were established by using infected cell DNA. DNA homology groupings of adenoviruses could be examined by spot hybridization, and restriction endonuclease cleavage patterns of viral DNAs could be obtained by Southern blot hybridization, by using infected cell DNA. The method was very sensitive and allowed the identification of the cleavage pattern of viral DNA of the inoculum by means of cell DNA extracted from infected cells with undetectable cytopathic effect (CPE). In ethidium bromide-stained gels without Southern blot hybridization, the restriction endonuclease cleavage pattern of viral DNA could be detected precisely in spite of background staining due to cellular DNA. The preparation of infected cell DNA used in these procedures was technically much easier than that of viral DNA. These methods require only a small number of infected cells and allow many isolates to be investigated with ease. PMID- 6278272 TI - Effect of mouse hepatitis virus infection on "natural killer cell" activity in nude mice. PMID- 6278273 TI - Response to aminoglutethimide after tamoxifen therapy in advanced breast cancer. AB - We describe sequential response to aminoglutethimide after eventual failure of treatment with tamoxifen in a patient with metastatic breast cancer. We suggest that aminoglutethimide is the next logical hormone therapy for such patients. PMID- 6278274 TI - Methyltestosterone-induced liver-cell tumours. PMID- 6278275 TI - Childhood cancer in offspring of two Wilms tumor survivors. AB - A search was made for cancers among offspring and siblings of 149 Connecticut born children with Wilms tumor reported to the Connecticut Tumor Registry during 1935 to 1973. Nasopharyngeal rhabdomyosarcoma developed in the daughter of a man with unilateral Wilms tumor that also affected his sister. Hodgkin disease developed in the daughter of a woman who had unilateral Wilms tumor. One other patient had a sibling with Wilms tumor and three had a sibling with other cancers (two Hodgkin disease, one testicular seminoma). The survey suggests an excess risk of other forms of cancer among the progeny and siblings of Wilms tumor patients. PMID- 6278276 TI - [Effect of phytobacteriomycin on biting fly larvae (Diptera, Simuliidae)]. PMID- 6278277 TI - [Antimony content of the thyroid of man and animals undergoing visceral leishmaniasis treatment with the organic compound of antimony, stibogluconate sodium]. PMID- 6278278 TI - [Effects of correction of eating behavior and metabolic consumption on diarrhea, subcutaneous tissue, growth and hemoglobin in 18 children with aspecific chronic diarrhea]. PMID- 6278279 TI - Acyclovir: an effective antiviral. PMID- 6278280 TI - Hormone receptors in breast cancer. PMID- 6278281 TI - [Malignant mixed tumor of the submandibular salivary gland]. PMID- 6278282 TI - Inactivation and reactivation of phosphoprotein phosphatase. AB - The catalytic subunit of phosphoprotein phosphatase (Mr = 35,000) is inactivated by phosphate compounds such as trimetaphosphate, PPi, and ATP. The inactivation of phosphoprotein phosphatase by these phosphate compounds is time- and concentration-dependent, is not reversed by dilution or gel filtration and is protected by Pi. A dissociation constant for the enzyme-trimetaphosphate complex and a rate constant for the reaction were calculated to be 4.6 x 10(-4) M and 0.29 min-1, respectively. The inactivation of phosphatase by PPi and ATP shows more complex kinetics than that by trimetaphosphate. The addition of EDTA to PPi and ATP exhibits more potent inactivation, even though EDTA alone does not inactivate phosphatase. This phosphoprotein phosphatase is not labeled by [gamma 32P]ATP. The inactivation of phosphatase by PPi or ATP can only be reversed by Mn2+ or Co2+, among all other metals or cationic compounds tried. The reactivation also requires sulfhydryl compounds. The effectiveness of sulfhydryl compounds follows the order: dithioerythritol greater than mercaptoethanol greater than cysteine. Glutathione was without effect. Metal analysis of the catalytic subunit did not reveal any significant amounts of Ca, Cd, Co, Cu, Fe, Mg, Mn, Ni, Sn, or Zn. Phosphoprotein phosphatase activity from zinc-deficient rat livers also eliminated the possibility of this phosphatase being a zinc metalloenzyme. Inactivation does not seem to be due to a loss of a critical metal ion. Other mechanisms for inactivation are presented. PMID- 6278283 TI - Histidyl-proline diketopiperazine: its biological role as a regulatory peptide. AB - Histidyl-proline diketopiperazine [cyclo(His-Pro)] is a metabolite of thyrotropin releasing hormone (TRH). This review summarizes the literature concerning cyclo (His-Pro) and, in addition, some studies dealing with TRH and other peptide that are considered of interest. The enzymes concerned with the metabolism of TRH are discussed. Distribution studies of peptides by immunological methods show that, while TRH is concentrated in synaptosomes, cyclo (His-Pro) is not, suggesting that cyclo (His-Pro) is not a classical neurotransmitter. Rat brain contains approximately three times as much cyclo (His-Pro) as TRH, mainly localized in the pituitary and hypothalamus. While the TRH is found in a free form, the cyclo (His Pro) is bound to a carrier of molecular weight approximately 70,000. While specific membrane receptors for TRH have been detected in pituitary cells, no such receptors for cyclo (His-Pro) have yet been found in brain or pituitary; however, there is a specific binding of cyclo (His-Pro) to adrenal cortex membranes. Both TRH and cyclo (His-Pro) have effects in the central nervous system or pituitary. These include effects on prolactin release, thermoregulation, CNS depression, stereotypic behavior and cyclic nucleotide levels. Possible mechanisms and interrelations of these effects are discussed. PMID- 6278285 TI - Activation of a phosphoprotein phosphatase in mouse 3T3 cells by insulin. PMID- 6278284 TI - Hormone-sensitive cAMP phosphodiesterase in liver and fat cells. PMID- 6278288 TI - Characterization of protein phosphorylation in acetylcholine receptor-enriched membrane preparations from Torpedo fuscomaculata. AB - When the acetylcholine receptor (AChR) from Torpedo is phosphorylated, ATP is found to bind non-covalently to the preparation. After correction is made for this binding of ATP, the phosphorylation reaction has a Km of 0,16 mM, a pH optimum of 8,6 and reaches maximal activity within 3 min. The intrinsic kinase activity is very specific for the AChR and does not phosphorylate either histones or phosvitin. The only amino acid to be phosphorylated is serine and cyclic GMP causes stimulation of the reaction. PMID- 6278287 TI - Restoration by cyclic AMP of the differentiated phenotype of chondrocytes from de differentiated cells pretreated with retinoids. AB - Parathyroid hormone (PTH) increases the cyclic AMP level in rabbit costal chondrocytes in culture. PTH, dibutyryl cyclic AMP (DBcAMP), and 8-bromo cyclic AMP (8-Br cAMP)induce ornithine decarboxylase (ODC) and expression of the differentiated phenotype of chondrocytes in this cell system. On the other hand, retinoids inhibit expression of the differentiated phenotype of chondrocytes. In the present study, the effects of PTH, DBcAMP, and 8-Br cAMP on rabbit costal chondrocytes pretreated with retinoids were examined. PTH did not increase the cellular cyclic AMP level in de-differentiated cells that had been pretreated with retinyl acetate or retinoic acid for three days, but it did increase the cyclic AMP level four days after removal of retinoids. PTH did not stimulate ODC activity or expression of the differentiated phenotype of chondrocytes in the de differentiated state. On the other hand, DBcAMP or 8-Br cAMP stimulated expression of the differentiated phenotype of chondrocytes even in de differentiated cells, as judged by morphological and histological changes of the cells and increase in glycosaminoglycan synthesis. Cyclic AMP analogues also induced ODC in these cells. PMID- 6278289 TI - [Determination of protein secondary structure from circular dichroism spectra. III. Protein-derived base spectra of circular dichroism for antiparallel and parallel beta-structures]. AB - Protein-derived basic CD spectra for alpha-helix, antiparallel and parallel beta structures, beta-bends and irregular form of proteins have been determined from the experimental CD spectra of six (myoglobin, lysozyme, ribonuclease A, papain, lactate dehydrogenase, subtilisin BPN') or seven (glyceraldehyde-3-phosphate dehydrogenase added) reference proteins and the analysis of the X-ray data. The secondary structures of thirteen proteins (seven reference and six additional ones) have been analysed using the basic CD spectra thus obtained. The data obtained have been compared with the results of the X-ray data analysis. It is shown that the accuracy of determination of the beta-structure and beta-bends contents using our basic CD spectra is about 2-3 times better than using the basic spectra reported by Chang et al. (Analyt. Biochem. 91, 13-31, 1978). PMID- 6278290 TI - [Comparison of ATP-pyrophosphate exchange reactions, catalyzed by two different "one-site" forms of tryptophanyl-tRNA-synthetase and by native enzyme]. AB - Kinetic parameters of the reaction of ATP-PPi exchange, catalysed by "one-site" form of beef pancreas tryptophanyl-tRNA-synthetase are determined. The one-site form was obtained by affinity labelling of one active centre of the enzyme with alkylated analog of tryptophan. The comparison of kinetic parameters of this one site form with the other one produced earlier by affinity labelling with analog of tryptophanyl-tRNA and the native enzyme was carried out. It was shown that the presence of tRNA improves the discrimination properties of the enzyme and diminishes the concentrations of "dead-end" complexes. Negative cooperativity in binding of two molecules of tryptophan is observed, which disappears in the presence of ATP, leading to an apparent synergistic effect. The molecule of tryptophan, which is present in the active centre, diminishes affinity of pyrophosphate. PMID- 6278291 TI - [Electron paramagnetic resonance of electron transport in photosynthetic systems. XI. Effects of photosynthetic control: dependence of the rate of electron transport on the energization of bean chloroplast thylakoid membrane]. AB - The kinetics of light-induced P700 redox transients in bean chloroplast was studied. It has been shown that the rate of electron transport decreased during few seconds of illumination of coupled chloroplasts without addition of ADP and inorganic phosphate. The evidence were obtained that there is a feedback inhibition of electron transport governed by the internal pH of thylakoid. This results in the overshoot in the kinetics of P700 redox transients induced by continuous actinic light. Under the phosphorylation condition (addition of Mg-ADP and inorganic phosphate) the effect of decreasing of the rate of electron transport between two photosystems was not observed. Addition of uncouplers (FCCP or gramicidine) also increased the steady-state rate of noncyclic electron transport. After adding only Mg-ADP (without phosphate) or Mg-ATP to coupled chloroplasts the effect of the light-driven inhibition of electron transport was observed as in the case of chloroplasts without any additions. We showed that the regulation for the electron transport rate was realized at the step of the plastoquinol oxidation by photosystem 1. Light-driven energization of the thylakoid membrane also leads to the the slowing of the reduction of spin label TEMPO. Evidences were obtained that TEMPO interacts with the semiquinone localized in the acceptor side of photosystem 2. From the comparative study of P700+ and TEMPO reduction by photosystem 2 we have concluded that there are two points of inhibitory action of DCMU localized at the acceptor and donor sides of photosystem 2. The mechanisms of photosynthetic control and the role of transmembrane proton gradient for energy transmission in chloroplasts are discussed. PMID- 6278292 TI - [Properties of protein released from mouse spleen nuclei under conditions of limited hydrolysis with DNase I and micrococcal nuclease]. AB - Proteins released by mouse spleen nuclei under conditions of limited hydrolysis with DNase I and micrococcal nuclease were compared. Analysis of these proteins by polyacrylamide gel electrophoresis in the presence of SDS revealed the essential similarity in the qualitative composition of released proteins. Characteristics of the most prominent component were studied in both cases and it was shown that this component is identical. It has a molecular weight of 25 500 according to electrophoresis data and 2300 as determined by equilibrium sedimentation. Amino acids composition and N-terminal amino acid were studied. It was shown that its N-amino acid is arginine. PMID- 6278286 TI - The sarcoplasmic reticulum Ca2+-ATPase. PMID- 6278293 TI - [EPR study of flavo- and ubisemiquinones in liver tissue]. AB - Several types of flavins and quinones having different parameters of magnetic relaxation have been detected in the mouse liver tissue by means EPR technique. The values of these parameters depend on the redox state of the respiratory chain meal-containing centres. This means that there is an interaction between the free radicals and the metal-containing paramagnetic centres. PMID- 6278294 TI - [Free-radical label--new approach to the study of dynamics of lipid systems]. AB - A new method of EPR-spectroscopy--the recombination of free radicals appearing as a result of indirect radiolysis of biological molecules after a low temperature irradiation--is applied to the study of molecular dynamics of phosphatidylcholine dimyristoyl in mass and in the structure of liposomes above and below the transition temperature. It was shown, that the mobility of lipid molecules in crystalline liposomes is higher than in the structure of liquid-crystalline liposomes. The addition of cholesterol in liposome membranes decreases the lateral molecular motion of lipids in crystalline and liquid-crystalline state, in the latter case the effect of cholesterol addition is more pronounced. The activation energy for the displacement of the fragments of lipid molecules and the lipid molecule as a whole was estimated, and it was shown, that lipid matrix possesses a high degree of heterogeneity. PMID- 6278295 TI - [Modification of Escherichia coli RNA polymerase by diethylpyrocarbonate. II. Binding and unwinding of double-stranded DNA]. AB - E. coli DNA dependent RNA polymerase was modified by diethylpyrocarbonate. Binding to a double-stranded DNA and unwinding of the DNA at the enzyme binding site by the modified enzyme were examined. It was found that RNA polymerase reversibly lost the ability to unwind DNA helix as well as the RNA synthetic activity when 9 to 11 histidyl residues of the enzyme were modified. In addition ot modification of the most reactive sulfhydryl or amino groups of the enzyme accompanying histidyl residues modification results in irreversible decrease of the salt concentration which is necessary to remove the enzyme from DNA cellulose column. Further modification of the less reactive sulfhydryl or amino groups leads to irreversible loss of the DNA binding ability and to the enzyme structure alteration. PMID- 6278296 TI - Cushing's syndrome. PMID- 6278297 TI - [Varicella-zoster-immunoglobulin as a safe prophylaxis of varicella (author's transl)]. AB - The occurrence of varicella in the course of immunosuppressive therapy is always a hazard to life. Up till now, passive immunization is the only method which provides reliable protection. We have had an immunoglobulin compound at our disposal during the last 5 years. This compound was obtained by screening a plasmapool for high-titre varicella antibodies, which were then fractionated. Varicella immunoglobulin was injected on 298 occasions to children incubated with varicella. There were no serious side effects after i.v. administration. These children were on polychemotherapy because of malignant diseases. Only 4 children developed varicella, 3 of them very mildly and none of them was later disabled due to varicella in any form. The protective value of this varicella immunoglobulin compound is much greater than any other known so far [16]. PMID- 6278298 TI - [Conjugated-estrogen treatment of excessively tall girls (author's transl)]. AB - 46 excessively tall girls, aged 10.5-15 years and with a mean height prediction of 185.5+/-3.3 cm were treated with 7.5 mg conjugated estrogens daily and 5 mg gestagens daily for 5-10 days. Treatment for 30 girls has been completed. Bone age was assessed according to Tanner et al. taking into account midparent height. Mean bone-age was 13.0+/-0.6 years at the beginning and 15.5+/-0.6 years at the end of treatment. Duration of therapy was 19.6+/- 6.8 months and mean reduction of predicted height in all patients was 4.1+/-2.4 cm. Bone age subgroups do not alter the results. Bone maturation was accelerated by treatment to 1.8 years/year with marked acceleration during the first six months. Growth velocity was low during the 6th-12th month. Increase of growth was 1.4+/-1.2 cm in the second period of therapy, representing 50% of the initial phase. Side-effects were minimal. The results are in agreement with those reported in other comparable series. PMID- 6278299 TI - [The paramedical approach and medical ethics (author's transl)]. AB - The fundamentally different approaches of modern science-based medicine and of those magic diagnostic and therapeutic procedures like iridiagnosis anthroposophical medicine, homeopathy, cell-therapy, neural-therapy, magnetic therapy and others, are analysed. Science-based medicine relies on the rules of the principle of causality, reproducibility, and predictability of its results, supported by its success. The dangers of modern medicine are its rationality and factualness which the doctor has to adapt individually to each patient. Otherwise he renders the patient susceptible to paramedical methods which make use of his suggestibility and need for comfort and consolation. Nothing is to be said against these methods as long as they are used as a psychotherapeutic vehicle, do no harm, and do not exploit the patient economically. If that would be the case, the patient must be protected against such quackery. PMID- 6278300 TI - [Chronic mycoses can be treated]. PMID- 6278301 TI - DNA-repair synthesis in ataxia telangiectasis lymphoblastoid cells. AB - The ability of a number of Epstein-Barr virus-transformed lymphoblastoid cells from ataxia telangiectasis (AT) patients to repair gamma-radiation damage to DNA was determined. All of these AT cells were previously shown to be hypersensitive to gamma-radiation. Two methods were used to determine DNA-repair synthesis: isopycnic gradient analysis and a method employing hydroxyurea to inhibit semiconservative DNA synthesis. Control, AT heterozygote and AT homozygote cells were demonstrated to have similar capacities for repair of radiation damage to DNA. In addition at high radiation doses (10-40 krad) the extent of inhibition of DNA synthesis was similar in the different cell types. PMID- 6278302 TI - Conservation of a variant-specific surface antigen gene in different trypanosome species and sub-species. AB - In Trypanosoma brucie brucie, T. b. rhodesiense, T. b. gambiense and T. evansi, the variant-specific antigen (VSA) genes are organized in families of related sequences, one of which is duplicated when expressed. Some of these VSA sequences appear to be conserved in the different species and sub-species: restriction mapping of isotypic genes of AnTat 1.8 VSA (from T. b. brucei) reveals extensive homology in T. b. rhodesiense, T. b. gambiense and T. evansi, although the genetic surrounding differs in each case. By contrast, the AnTat 1.1 sequence (also from T. b. brucei) appears to be absent from T. b. gambiense DNA. PMID- 6278304 TI - Current concepts in otolaryngology: malignant neoplasms of the minor salivary glands. PMID- 6278303 TI - Deficiency of a granulocyte-membrane glycoprotein (gp150) in a boy with recurrent bacterial infections. PMID- 6278305 TI - Missing hormone in glucocorticoid-suppressible hypoaldosteronism. PMID- 6278306 TI - Erythrocyte sodium-potassium-ATPase and obesity. PMID- 6278307 TI - Epstein-Barr virus-induced B-cell lymphoma after renal transplantation: acyclovir therapy and transition from polyclonal to monoclonal B-cell proliferation. PMID- 6278308 TI - Opportunistic infections and Kaposi's sarcoma in homosexual men. PMID- 6278309 TI - Congenital cytomegalovirus infection: The relative importance of primary and recurrent maternal infection. AB - We studied the incidence of primary and recurrent cytomegalovirus infection in 3712 pregnant women--2698 of middle to high income and 1014 of low income--to determine whether there were differences in the effects on the fetus. In the higher-income group, 1203 women (45 per cent) did not have antibodies to cytomegalovirus and were therefore susceptible to primary infection, as compared with 179 women (18 per cent) of low income. Congenital infection occurred more often (1.6 vs. 0.6 per cent) in infants in the low-income group. In this group it was associated with recurrent maternal infection more often (in 82 per cent) than with primary maternal infection, whereas in the upper-income group, it was associated with primary maternal infection in half the cases. Altogether, there were 32 cases of congenital cytomegalovirus infection - 16 in each group. Whereas primary maternal infection resulted in fetal infection in only half the cases, it was more likely to ge associated with clinically apparent disease than was recurrent infection. When these cases were combined with 28 cases of congenital infection retrospectively identified at other prenatal clinics, five of 33 infected infants born after primary maternal infection had clinically apparent disease, as compared with none of 27 born after recurrent maternal infection. We conclude that congenital cytomegalovirus infection resulting from primary maternal infection is more likely to be serious than that resulting from recurrent infection, and is more likely to occur in upper socioeconomic groups. PMID- 6278310 TI - Cmv immunity: imperfect but protective. PMID- 6278311 TI - Regulation of metallothionein--thymidine kinase fusion plasmids injected into mouse eggs. AB - A plasmid was constructed with the promoter/regulatory region of the mouse metallothionein-I gene fused to the structural gene of herpesvirus thymidine kinase. When mouse eggs were microinjected with this plasmid and incubated with cadmium (a natural inducer of metallothionein gene transcription) thymidine kinase activity increased approximately 10-fold compared with control eggs not exposed to cadmium. Analysis of a set of deletion mutants revealed that the minimum sequence required for cadmium regulation lies within 90 nucleotides of the transcription start site. PMID- 6278312 TI - A viral polymerase involved in recognition of influenza virus-infected cells by a cytotoxic T-cell clone. PMID- 6278313 TI - Selective blockage of voltage-dependent K+ channels by a novel scorpion toxin. PMID- 6278314 TI - Triphosphoinositide increases glycoprotein lateral mobility in erythrocyte membranes. PMID- 6278315 TI - Growth factors adherent to cell substrate are mitogenically active in situ. PMID- 6278316 TI - Selective effect of feline leukaemia virus on early erythroid precursors. PMID- 6278318 TI - Nanomolar concentrations of extracellular ATP activate membrane Ca channels in snail neurones. PMID- 6278317 TI - High-affinity ouabain binding site and low-dose positive inotropic effect in rat myocardium. PMID- 6278319 TI - Genetic diagnosis of the fetus. PMID- 6278320 TI - Genome instability in a region of human DNA enriched in Alu repeat sequences. PMID- 6278321 TI - beta-Endorphin is processed differently in specific regions of rat pituitary and brain. PMID- 6278322 TI - Nuclear localization and DNA binding of the transforming gene product of avian myelocytomatosis virus. PMID- 6278323 TI - Regulation of DNA ligase activity by poly(ADP-ribose). PMID- 6278324 TI - Single Ca2+-activated nonselective cation channels in neuroblastoma. PMID- 6278325 TI - DNA sequence homology and chromosomal deletion at a site of SV40 DNA integration. PMID- 6278326 TI - Self-excitation of olfactory bulb neurones. PMID- 6278327 TI - Opioid peptides induce reduction of enkephalin receptors in cultured neuroblastoma cells. PMID- 6278328 TI - Influence of ionic current on Na+ channel gating in crayfish giant axon. PMID- 6278329 TI - Nerve impulses increase the phosphorylation state of protein I in rabbit superior cervical ganglion. PMID- 6278330 TI - [Metabolism during shock and its therapeutic regulations]. AB - During shock the organism reacts by means of primarily useful autoregulation mechanisms which, however, in excess become useless and at least harmful. Therapeutically it is necessary now, to partly break through these mechanisms going along with substitution of certain necessary substances and besides that in part to support the organism's own concept. PMID- 6278331 TI - Effects of hydrogen fluoride on benzo[a]pyrene and dimethylnitrosamine metabolism in rats. PMID- 6278332 TI - 3H-Noradrenaline and 3H-5-hydroxytryptamine release from rat brain slices and its presynaptic alpha-adrenergic modulation after long-term desipramine pretreatment. AB - The electrically-evoked release of 3H-noradrenaline from superfused cortex and hippocampus slices was strongly enhanced (by about 50%) after long-term (4 weeks) pretreatment with desipramine (10 mg/kg, twice daily). The release-enhancing effect of the alpha-receptor antagonist phentolamine was significantly reduced but the inhibitory effects of exogenous noradrenaline and clonidine on 3H noradrenaline release were virtually unchanged after chronic desipramine treatment. The K+ -induced release of 3H-noradrenaline from superfused synaptosomes obtained from rats pretreated with desipramine was about 25% higher than that from synaptosomes of control animals. However, noradrenaline inhibited the K+ -induced synaptosomal 3H-noradrenaline release to the same extent (viz. by about 55%) in both cases. The release of 3H-5-hydroxytryptamine induced by 26 mM K+ from cortex and hippocampus slices was not affected by chronic pretreatment with desipramine. In addition, no change was observed in the inhibitory effect of noradrenaline o 3H-5-hydroxytryptamine release. It is concluded that long-term pretreatment with desipramine leads to selective changes in the basic mechanism(s) of noradrenaline release rather than to changes in the sensitivity of presynaptic alpha-adrenoceptors. PMID- 6278334 TI - [Are scintigraphy and echography of the thyroid supplementary examination methods?]. PMID- 6278333 TI - In vivo and in vitro desensitization of nicotinic acid-induced adipocyte adenylate cyclase inhibition. AB - The influence of pretreatment of hamsters and of intact hamster adipocytes with the heterocyclic carboxylic acid, 3-carboxy-5-methylpyrazole (CMP) was studied on the inhibitory effects of various antilipolytic agents on adenylate cyclase in membrane preparations. In vivo pretreatment of hamsters for 3 days with the potent antilipolytic agent, CMP (40 mg/kg b.w., twice daily), markedly reduced the inhibitory potencies of CMP itself and of nicotinic acid on the adenylate cyclase. In contrast, enzyme inhibition by PGE1 remained unchanged. Increase in adenylate cyclase activity due to the stimulatory hormone, ACTH, the basal activity were enlarged in adipocyte membranes from CMP-pretreated hamsters compared to controls. In vitro pretreatment of intact hamster adipocytes with CMP at a maximally effective concentration (300 microM) caused a time-dependent decrease in adenylate cyclase inhibition by CMP itself; upon pretreatment for 120 min, inhibition was diminished by about 50%. Identical decreases in the inhibitory potencies were observed for nicotinic acid and for 3-carboxy-5 methylisoxazole, another carboxylic acid. In contrast, inhibitions of the adipocyte adenylate cyclase by PGE1, adrenaline and N6 -phenylisopropyladenosine were not changed by the in vitro pretreatment with CMP. The data indicate that adipocyte adenylate cyclase inhibition can desensitize following both in vivo and in vitro administration of an inhibitory agent. The desensitization process and the specificity observed give further support to the idea that inhibition of adipocyte adenylate cyclase by nicotinic acid is mediated by a membrane receptor and, additionally, suggest that nicotinic acid and the heterocyclic carboxylic acids studied act via a common receptor. PMID- 6278336 TI - [Changes in the postsynaptic responses of spinal cord motor neurons against a background of rhythmic stimulation of the locus coeruleus]. AB - Influences of locus coeruleus rhythmical stimulation on the postsynaptic reactions of spinal motoneurons were studied in chloralose anesthetized cats. IPSPs evoked by flexor reflex afferent (FRA) stimulation were inhibited, but EPSPs evoked by FRA and PSPs evoked by low-threshold muscle afferents stimulation were unchanged under these conditions. The inhibition of IPSPs disappeared in reserpine-pretreated animals. Both IPSPs and EPSPs evoked by FRA stimulation were diminished also during rhythmical stimulation of nucleus reticularis pontis oralis. However this effect was not of monoaminergic nature because it was present in reserpine-pretreated animals. PMID- 6278335 TI - [A patient with (South American) mucocutaneous leishmaniasis recurring after treatment with co-trimoxazole]. PMID- 6278337 TI - [Glial origin of the slow negative potential of the direct cortical response: microelectrode study and mathematical analysis]. AB - Slow negative potential of a direct cortical response is similar in configuration, time-course and reaction to repeated stimuli to depolarization of the cortical glial cells and differs from IPSP of the cortical neurons. According to data of digital spectral (frequency) analysis, slow negative potential is based on the glial component formed by summing up the constituents, which coincide with glial depolarization within a constant factor. The neuronal component, whose contribution is comparatively much smaller, is an indirect result of IPSP. PMID- 6278338 TI - [Theoretical description of the calcium channel in the spinal ganglion neurons of the rat]. AB - The surface charge density (sigma'0 and the constant of Ca2+ binding with the charged surface groups of the outer side of the membrane (KCa) are calculated on the basis of experimentally determined values of the current-voltage characteristic shifts for calcium currents in the membrane of rat spinal ganglia neurons. The obtained values are as follows: sigma'0 = 0.15 +/- 0.05 e/nm2 and KCa = 70 +/- 10 1/ml. The energy profile of the calcium channel in the membrane of investigated neurons was calculated for Ca2+, Ba2+ Cd2+ Mn2+ Co2+ ions and verapamil using a three-barrier model. It is shown that the value of calcium current is determined mainly by the potential hole depth which corresponds to the outer binding centre of the calcium channel. On the basis of the data obtained a conclusion is made that the outer binding centre contains only one carboxylic group. PMID- 6278339 TI - [Sintered calcium hydroxyapatite implants, transmucosally inserted. An experimental animal study]. PMID- 6278340 TI - [Gliomas induced in mammals by Rous sarcoma virus (RSV) (author's transl)]. AB - Studies concerning the induction of gliomas in a variety of animal species by an oncogenic avian virus (Rous sarcoma virus) were presented. Experimental conditions (age of inoculation, doses of virus) were reviewed. Preliminary attempts to localize viral antigens in the tumors in vivo using labelled antibodies were discussed. PMID- 6278341 TI - [Comparative ultrastructure of human gliomas and experimental gliomas induced by Rous sarcoma virus (RSV) (author's transl)]. AB - A brief review of earlier results concerning the ultrastructure of 20 gliomas induced by intracerebral inoculation of Rous sarcoma virus in the dog emphasizes the similarity of certain canine and human histological types. Since no sign of the viral induction is recognizable for certain in the experimental tumors and since the large size of the animal authorizes the use of the same radiological detection techniques as in Man these viral induced tumors represent a remarkable experimental model. PMID- 6278342 TI - Pharmacokinetics of inhaled gases and vapors. AB - Physiological and biochemical factors determine the kinetic patterns of uptake, distribution, metabolism, and elimination of inhaled gases and vapors. For metabolically inert gases, true equilibrium is achieved after appropriately long exposures and the overall shape of the time-course curves of uptake and elimination should be concentration-independent. At equilibrium, achieved internal concentrations will be linearly related to exposure concentration. The rate of approach to equilibrium depends on blood flows for poorly soluble chemicals and on alveolar ventilation for soluble gases and vapors. For metabolized gases and vapors, steady-state is achieved where net pulmonary uptake replaces chemical removed from the circulation by metabolism. The shape of time course curves for soluble, well-metabolized chemicals will be concentration dependent, and steady-state blood:gas concentration ratios will be complexly related to inhaled concentrations. Physiological models of inhalation pharmacokinetics allow extrapolation of results in one species to untested species, based on knowledge of changing physiology between animal-to-animal. Two potential applications of interspecies extrapolation are discussed. PMID- 6278343 TI - The neurobehavioral toxicity of trichloroethylene. AB - Trichloroethylene is an industrial solvent used primarily in degreasing operations with some use as an anesthetic agent as well. The primary route of exposure is inhalation and the central nervous system effects consist of headache, nausea, sleepiness, burning of the eyes. Human cases of intoxication have been associated with trigeminal neuropathy, however this is probably caused by a breakdown product dichloroacetylene. Fatal exposures may be the result of cardiac failure. Chronic exposure in industrial settings may cause alterations in a variety of behavioral parameters such as reduced memory and intellectual functioning. Experimental human exposures reveal fatigue and sleepiness effects and possible alterations in reaction time, but no deterioration in performance on manual dexterity tasks up to 300 ppm exposures. Animal experiments using acute exposures generally fail to reveal behavioral effects at concentrations below 1000 ppm, with a range of 75-2000 ppm. Cessation of exposure results in rapid behavioral recovery with no residual behavioral deficits. Exposure of dogs to 3000 ppm chronically results in severe cerebellar pathology, with no trigeminal nerve damage. No neurochemical effects of exposure have been documented. The neurobehavioral literature on the toxic effects of trichloroethylene is fragmented and poorly documented suggesting that more and better quality work is needed to understand the potential toxicity of this compound. PMID- 6278344 TI - Hexacarbon neurotoxicity. AB - The neurotoxicity of hexacarbon solvents has become apparent only recently, but an extensive literature has already developed as a result of the clinical and epidemiological implications of the human disease, and the advantages of small animal models of hexacarbon neurotoxicity in investigation of experimental neuropathies. This review selectively summarizes the literature, focusing on clinical manifestations and pathological alterations in the peripheral nerves. The clinical manifestations are dominated by a distal, symmetrical, sensorimotor polyneuropathy. Pathological changes include distal axonal degeneration and formation of neurofilament-filled axonal swellings in early or moderately severe exposure. A late manifestation in severely affected experimental animals is recurrent demyelination and remyelination. PMID- 6278345 TI - Immunoreactive beta-endorphin in the plasma, pituitary and hypothalamus of young and old male rats. AB - Immunoreactive beta-endorphin (IR-beta-ENDO) was measured in the plasma, pituitary, and hypothalamus of young (3-5 mo.) and old (19-23 mo.) male Sprague Dawley rats, using a specific radioimmunoassay. Plasma IR-beta-ENDO in old male rats (3.44 +/- 0.54 ng/ml) was more than three times higher than values observed in young male rats (1.00 +/- 0.10 ng/ml). Pituitary content and concentration of IR-beta-ENDO also were significantly greater in the old (5.85 +/- 0.51 microgram/gland and 1.17 +/- 0.10 microgram/mg protein) than in the young (3.53 +/- 0.29 microgram/gland and 0.78 +/- 0.06 microgram/mg protein) male rats. The content of IR-beta-ENDO in the hypothalamus of old and young rats was nearly the same (43.45 +/- 2.47 and 49.88 +/- 6.35 ng/hypothalamus, respectively), whereas the concentration of IR-beta-ENDO in the hypothalamus of the old male rats (3.89 +/- 0.25 ng/mg protein) was approximately 50% lower than that observed in the young male rats (7,80 +/- 0.85 ng/mg protein). These changes in plasma, pituitary, and hypothalamic IR-beta-ENDO may contribute to the increase in prolactin and decrease in gonadotropins observed in old male rats, since beta ENDO administration is known to produce these effects on prolactin and gonadotropin secretion. PMID- 6278346 TI - Functional interaction between benzodiazepine and GABA recognition sites in aged rats. AB - The present study was undertaken to explore whether there may be age-related changes in benzodiazepine binding and in the functional interaction between GABA and benzodiazepine recognition sites. Data indicate an increase in benzodiazepine binding sites with age. Moreover the functional interactions between GABA and benzodiazepine receptor sites are differentially affected by aging. GABA is less active in enhancing benzodiazepine binding in the older animals because of the loss of GABA receptors, and Diazepam may be more active in enhancing GABA receptor binding in the aged animals because there are more benzodiazepine receptors in this group. An understanding of the relevance of the apparent alteration in coupling between GABA and benzodiazepine receptors must permit a better definition of the behavioral manifestation of their biochemical phenomenon. PMID- 6278349 TI - [Intraoperative radiotherapy and conformation radiotherapy for malignant brain tumors (author's transl)]. PMID- 6278350 TI - The possible adjuvant role of bordetella pertussis and pertussis vaccine in causing severe encephalopathic illness: a presentation of three case histories. AB - The clinical and some laboratory details of three children who had severe neurological sequelae after either infection with Bordetella pertussis or immunisation with diphtheria, tetanus and pertussis vaccine and oral polio vaccine are reported. Each of these patients had had a recent or concurrent viral illness. The severity of their encephalopathic illness may have been due to an adjuvant role of B. pertussis or a component of the vaccines they received. PMID- 6278347 TI - Detergent solubilization and hydrophobic chromatography of rat brain phosphatidylinositol kinase. AB - Rat brain microsomal phosphatidylinositol kinase activity was maximally activated in the presence of either 3 mM sodium deoxycholate, 2% Triton-X-100, or 30-40 mM octylglucoside. Among these detergents, 1% Triton-X-100 was most effective in solubilizing the enzyme, and after treatment with this agent, 100% of the activity was recovered in the high speed supernatant. Octylglucoside solubilized 40% of the enzyme at concentrations below its critical micelle concentration of 25 mM and up to 80% at higher levels. Solubilized phosphatidylinositol kinase failed to absorb to adenosine nucleotide affinity resins. However, when the Triton-X-100 extract was chromatographed on an uncharged hydrophobic resin, consisting of dodecyl chains attached to Sepharose 4B by ether bonds, nearly all the enzyme activity was retained, and from 44-85% could be eluted with 8 mM sodium deoxycholate. Solubilization followed by hydrophobic chromatography resulted in several-fold purification of phosphatidylinositol kinase and may have disrupted interactions of the enzyme with other hydrophobic proteins sufficiently to allow its substantial purification by conventional or affinity chromatography techniques. PMID- 6278348 TI - Characteristics of the norepinephrine-stimulated phosphatidylinositol turnover in rat pineal cell dispersions. AB - Dispersed rat pineal cells can be used for the study of the phosphatidylinositol effect. The response to ( - )-norepinephrine of the incorporation of 32Pi into phospholipids is linear with time and cell concentration, stereospecific, and mediated through alpha-1-adrenergic receptors. Na+ in the incubation medium is obligatory for labeling of phosphatidylinositol and phosphatidylcholine by 32P. In the absence of K+, incorporation of 32P is drastically lowered and no stimulation by norepinephrine occurs. Rb+ can replace K+. Omission of Ca2+ or substitution with Sr2+ preferentially lowers incorporation of radioactivity into phosphatidylcholine. Mg2+ is not required for basal or stimulated labeling. PMID- 6278351 TI - Acute changes in cyclic AMP levels of certain brain areas following intraventricular injection of renin in rats. AB - Actions of the central angiotensin system on brain cAMP concentrations were studied after intraventricular injection of renin. Biosynthesis of endogenous brain angiotensin II was stimulated by 0.1 U renin and the blood pressure was elevated 40 min after the application. Stimulation of the brain angiotensin system affected cAMP metabolism in several brain areas, mostly expressed as decreased cAMP levels. Significant decreases in cAMP concentrations were found in the cerebral cortex, hippocampus, basal ganglia, various hypothalamic nuclei, substantia nigra, central grey matter and in the locus coeruleus. The unchanged cAMP levels in the nucleus of the solitary tract might indicate that intraventricular injection of renin induced an acute blood pressure elevation which is not associated with changes in the cAMP system in the primary baroreceptor area. PMID- 6278352 TI - Stereospecific accumulation of dihydromorphine and naltrexone by corpus striatal slices of morphine-dependent mice. AB - Stereospecific accumulation of [3H]dihydromorphine and [3H]naltrexone by striatal slices from morphine-dependent mice was examined in Krebs-Ringer bicarbonate medium. Striatal slices showed a saturable and stereospecific accumulation of both [3H]ligands. The accumulation constant of naltrexone, determined by Wilkinson's analysis, was significantly decreased in both morphine-dependent mice and dependent mice abruptly withdrawn for 6 hr. The maximal accumulation of naltrexone was not changed in withdrawn mice, but decreased in dependent mice. This could be due to the high concentration of residual morphine in the slices. There were no significant differences in the accumulation constant or maximal accumulation of dihydromorphine among the striatal slices from control, dependent and withdrawn mice. These data indicate that in morphine-dependent mice, there is an increased affinity of the opioid receptors for the narcotic antagonist, naltrexone but not for the agonist, dihydromorphine. PMID- 6278353 TI - Central excitatory properties of delta 9-tetrahydrocannabinol and its metabolites in iron-induced epileptic rats. AB - The effects of delta 9-tetrahydrocannabinol (delta 9-THC), two of its metabolites, 8 beta-hydroxy-delta 9-THC and 11-hydroxy-delta 9-THC, and cannabidiol were comparatively studied by means of an iron-induced cortical focal epilepsy in conscious rats with chronically implanted electrodes. delta 9 Tetrahydrocannabinol produced depression of the spontaneously firing epileptic focus, excitatory behavior, generalized after-discharge-like bursts of epileptiform polyspikes and frank convulsions. The pharmacological profiles of the two metabolites differed from that of the parent compound: 11-Hydroxy-delta 9 THC did not precipitate convulsions, but it did elicit all the other effects of delta 9-THC; the 8 beta-hydroxy derivative, on the other hand, exerted only two delta 9-THC-like effects; that is, it evoked polyspike bursts and convulsions. In contrast, cannabidiol, even in large doses (100 mg/kg) was devoid of all the effects of delta 9-THC. Furthermore, pretreatment with cannabidiol markedly altered the responses to delta 9-THC in the following ways: focal depression was partially blocked, polyspike activity was enhanced and convulsions abolished. Phenytoin pretreatment elicited similar effects, but it failed to block the delta 9-THC-induced convulsions. In general, the cannabinoids exhibit a wide spectrum of CNS effects ranging from focal depression to convulsions; specifically, however, the pharmacological profile of each agent can differ markedly; for example, the convulsant properties of delta 9-THC are not a universal characteristic of this class of drugs. PMID- 6278354 TI - Chronic benzodiazepine treatment increases [3H]muscimol binding in mouse brain. AB - High affinity [3H]muscimol receptors were analyzed in the forebrain and cerebellum of mice that had been treated on a chronic regimen with chlordiazepoxide and clonazepam. In the forebrain, only clonazepam induced an increase in the number of muscimol binding sites 2 hr after drug treatment. In the cerebellum, both the chlordiazepoxide and clonazepam-treated animals showed an increased number of muscimol binding sites 2 hr after drug treatment. At 26 hr after drug treatment, only the clonazepam-treated animals still revealed an increased number of cerebellar muscimol binding sites. Chronic benzodiazepine administration, therefore, induced an increase in the apparent number of high affinity muscimol binding sites in both the forebrain and cerebellum. PMID- 6278355 TI - Chronic clonazepam administration induces benzodiazepine receptor subsensitivity. AB - Clonazepam and chlordiazepoxide were administered chronically in increasing doses for three weeks in two different strains of mice. Forebrain [3H]diazepam binding was assayed in groups of mice sacrificed at 2, 26, 50 hr and 10 days following the last dose. Scatchard and single point analyses revealed a significant decrease in the number of [3H]diazepam binding sites [Bmax] which persisted for at least two days following chronic clonazepam treatment. The Bmax changes observed following chlordiazepoxide treatment were less pronounced than those elicited by clonazepam. No significant changes in receptor binding affinity (Kd) were detected with either drug. In the clonazepam-treated animals, Bmax values returned to normal by day 10 after drug treatment. Chronic benzodiazepine administration therefore induced a decrease in the apparent number of benzodiazepine binding sites in the mouse forebrain. The magnitude and duration of the observed subsensitivity appears to depend on the potency of the administered benzodiazepine. PMID- 6278356 TI - Specific proconvulsant action of an imidazobenzodiazepine (RO 15-1788) on isoniazid convulsions. PMID- 6278357 TI - Differences in central effects of beta-endorphin and enkephalins: beta-endorphin. A potent psychomotor stimulant. AB - The endogenous opiate-like peptides, beta-endorphin, methionine- and leucine enkephalin have been investigated in unanaesthetized cats after intracerebroventricular injection. beta-Endorphin produced marked and prolonged psychomotor stimulation (restlessness, apprehension, looking around, vacant stare and impelling locomotion), accompanied by pupillary dilation and tremor which was prevented by nalorphine. In contrast to beta-endorphin, the enkephalins did not cause affective behavioural phenomena. However, the enkephalins evoked transient and inconsistent vomiting which was also prevented by nalorphine. It is apparent, therefore, that morphinomimetic brain peptides are involved in at least two functions in the central nervous system: beta-endorphin subserves the mediation of a long-lasting psychomotor stimulation, while the enkephalins mediate vomiting of a transient character. PMID- 6278358 TI - Elevation of Met5-enkephalin and beta-endorphin hypothalamic content in rats receiving anorectic drugs: differences between D-fenfluramine and D-amphetamine. AB - D-Fenfluramine, an anorectic that releases serotonin (5-HT), repeatedly injected in rats (15 mg/kg per day) enhanced the met5-enkephalin and beta-endorphin content of the hyhpothalamus. The onset of this effect was slow, reaching a peak at 5 days; the increase in beta-endorphin gradually declined toward control level while the drug was still being administered although that of met-enkephalin persisted for 15 days. The elevation of the opioid peptide content of the hypothalamus was temporally associated with a slowing in the rate of body weight increase. A transient, small, increase in striatal met-enkephalin content was also induced by repeated D-fenfluramine injections; however the met-enkephalin content of frontal cortex, hippocampus and brainstem was not affected. A modification of the beta-endorphin content of hypothalamus was not seen after acute injection of D-fenfluramine or D-amphetamine but an increase was observed during repeated treatment with D-fenfluramine. Repeated injections of D amphetamine for 5 days (4.5 mg/kg per day) failed to increase either the met enkephalin or the beta-endorphin content of the hypothalamus. These data suggest that the anorexia elicited by repeated injections of D-fenfluramine but not that elicited by D-amphetamine, includes a participation by hypothalamic and beta endorphin stores. PMID- 6278359 TI - Effects of acute and chronic haloperidol treatment on the concentrations of immunoreactive beta-endorphin in plasma, pituitary and brain of rats. AB - Haloperidol (0.03-3 mg/kg) dose-dependently increased the plasma concentrations of immunoreactive beta-endorphin (ir-beta-END) without changing the concentrations of ir-beta-END in the adenohypophysis or the neurointermediate pituitary of rats. Elevated levels of ir-beta-END in plasma were also found after chronic treatment with haloperidol (1 mg/kg per day) for 3 weeks. The long-term treatment, however, caused a selective increase (60%) of the ir-beta-END concentrations in the neurointermediate pituitary without changing that in the adenohypophysis. Significantly elevated levels of ir-beta-END were also found in the hypothalamus and the septum. Gelchromatographic separation of the plasma components reacting with the beta-endorphin antiserum revealed that both the acute and chronic haloperidol treatment increased the plasma concentrations of immunoreactive material with the molecular size of beta-endorphin and beta lipotropin. This indicates that, at least, part of ir-beta-END released by haloperidol into plasma is of adenohypophyseal origin, since no material with the molecular size of beta-lipotropin has been found in the neurointermediate pituitary. PMID- 6278360 TI - Adrenergic receptors in rat spinal cord. AB - Radioligand binding assays were used to demonstrate the presence of alpha 1, alpha 2 and beta receptors in rat spinal cord. Specific and saturable binding was exhibited for [3H]-WB 4101 (alpha 1), [3H]-aminoclonidine (alpha 2) and [3H] dihydroalprenolol (beta). Binding was of high affinity and the total number of binding sites (Bmax) were: alpha 1, 66.5 +/- 1.0 fmol/mg protein; alpha 2, 20.0 +/- 0.6 fmol/mg protein; beta, 10.2 +/- 0.3 fmol/mg protein. The data confirms the existence of adrenergic receptors in spinal cord and provides further evidence of the role of catecholaminergic neurons in regulating spinal cord physiology. PMID- 6278361 TI - Isotachophoresis quantitation of subfractions of multiple sclerosis intra-blood brain barrier IgG synthesis modulated by ACTH and/or steroids. PMID- 6278362 TI - Antibodies to coronaviruses OC43 and 229E in multiple sclerosis patients. AB - Multiple sclerosis (MS) and matched control sera had similar antibody titers to coronaviruses OC43 and 229E when tested by a radioimmunoassay method. In contrast, cerebrospinal fluid from MS patients contained coronavirus antibodies more frequently and in higher titers than matched controls. Intrathecal antibody synthesis to OC43 and 229E viruses was detected in 41% (9/22) and 26% (7/27) of MS patients, respectively, but was not found in any of the neurologic control patients. This intrathecal antibody synthesis may mean that coronaviruses play an etiologic or pathogenic role in MS. Alternatively, intrathecal synthesis of coronavirus antibodies may be but part of a generalized and variable intrathecal antibody synthesis that is typical for MS patients. PMID- 6278363 TI - Fabry disease: significance of ultrastructural localization of lipid inclusions in dermal nerves. AB - An ultrastructural examination of intradermal nerve fibers in Fabry disease revealed signs of lipid accumulation and small unmyelinated nerve fiber degeneration. Many axons were swollen, and their internal organelles were lost. In several damaged axons, dense inclusions, probably lipid, were observed. No lipid inclusions were found in Schwann cells, which may indicate that they utilize different metabolic processes or are impervious to ceramide trihexoside. It is hypothesized that Schwann cells and myelin sheaths act as a metabolic barrier protecting the larger myelinated fibers. Lacking this barrier, the smaller unmyelinated fibers are more susceptible to lipid infiltration. This view may explain the small fiber neuropathy in Fabry disease. PMID- 6278364 TI - Abnormalities of cerebral oxidative metabolism in animal models of Parkinson disease. AB - Abnormalities of cerebral oxidative metabolism were investigated in "animal models" of Parkinson disease by in situ optical measurements of local cerebral blood volume and cytochrome oxidase redox shifts in rats two weeks after unilateral 6-hydroxydopamine lesions of the substantia nigra with or without interruption of ascending noradrenergic pathways. The data demonstrate oxidative metabolic dysfunction of ipsilateral cerebral hemispheres caused by lesions that involve both dopaminergic and noradrenergic systems but not when dopaminergic neurons only are affected. We speculate that the dementia of Parkinson disease may be more prevalent when degeneration of catecholaminergic systems is widespread and not restricted to the dopaminergic system. PMID- 6278366 TI - Peripheral neuropathy in epileptics. PMID- 6278365 TI - Disulfiram neuropathy: a neurofilamentous distal axonopathy. AB - Disulfiram is used to treat alcoholism and is known to cause peripheral neuropathy: few reports of biopsied human nerves have revealed axonal degeneration and loss of myelinated fibers. We studied a 22-year-old woman with severe sensorimotor neuropathy following treatment with disulfiram for 6 months. Histologic studies of the sural nerve revealed a neurofilamentous axonopathy with rare enlarged axons distended by neurofilaments. Disulfiram is converted enzymatically to carbon disulfide, which causes neurofilamentous distal axonopathy in animals. Similar changes in human nerve after disulfiram administration suggest that carbon disulfide is the toxic agent. PMID- 6278367 TI - [Needle aspiration in the cytodiagnosis of breast neoplasms]. PMID- 6278368 TI - [Neuroendocrine study in 2 children with headache treated with lisuride]. PMID- 6278369 TI - [Latent neuropathy in rheumatoid arthritis. Electrophysiological study of 45 cases]. AB - Thirty (88.2%) of thirty four patients with rheumatoid arthritis showed evidence of latent neuropathy, as judged by the following tests: measurements of motor and sensory conduction velocity; analysis of single motor units at various sites and under different conditions. All patients demonstrating electrophysiological signs of involvement of nervous functions showed no clinical signs of peripheral neuropathy. On the basis of the present results it is proposed that neurophysiological alterations could depend on a widespread (immunologically mediated?) injury of the axonic membrane. PMID- 6278370 TI - The CAMP test performed by using Staphylococcus aureus sphingomyelinase (beta haemolysin) and Clostridium perfringens lecithinase. AB - The CAMP test was performed by employing Staphylococcus aureus beta-haemolysin (sphingomyelinase) and Clostridium perfringens alpha-toxin (lecithinase) for identification of group B streptococci on blood agar, using blood from different origin. Partial purification of Cl perfringens alpha-toxin was carried out by means of sheep erythrocytes. With the toxin preparations described positive CAMP reaction was obtained only on sheep blood agar. False positives with regard to group A streptococci could not be avoided by either of the methods. PMID- 6278371 TI - NITA's I.V. standards/CDC's I.V. guidelines. PMID- 6278373 TI - Effects of colchicine on myelination of rabbit optic nerve: a biochemical study. AB - The effects of intravitreous injections of colchicine on the myelination of the optic nerve were studied in rabbits at different times during development using biochemical and morphological techniques. Myelin basic protein concentrations and 2',3-cyclic nucleotide 3'-phosphodiesterase activities were affected in a similar way by colchicine treatment and reflected the degree of myelination of the tissue. During early development, colchicine produced axonal degeneration and secondary demyelination (Wallerian degeneration). Later, axonal and myelin abnormalities were more variable. Some demyelination was observed amd myelin formation also may have been inhibited. Thus, the effect of colchicine was proportional to the degree of optic nerve and retinal maturation (the youngest being the most sensitive) and, to a lesser extent, to the dosage, Under conditions used for this study, no remyelination was observed. In adult animals, no lesions could be detected histologically one to four months after injection of colchicine, Thus, intravitreous administration of colchicine is a useful chemical technique for producing Wallerian degeneration in optic nerves of young developing rabbits. PMID- 6278372 TI - Cholinergic actions of false neurotransmitters: acetyldiethylcholine. AB - Acetyldiethylcholine (AcDECh), a false transmitter at cholinergic synapses, binds to central muscarinic receptors with 14-fold lower affinity than acetylcholine (AcCh). The properties of this binding, including the limited extent of regional specificity and binding activation by N-ethylmaleimide, are those associated with weak agonists and antagonists. Intracerebroventricular injection of AcDECh or AcCh produces an increase in arterial blood pressure which is blocked by prior administration of atropine (10 nmol). While AcDECh is 17-fold less potent tha AcCh in producing the pressor response, the maximum pressure changes and time courses are comparable. AcDECh also binds to nicotinic receptors of Torpedo electric organs thereby inducing conformational changes in the receptor-ion channel complex that are associated with postsynaptic activation. In these nicotinic actions, however, AcDECh is 320-fold less active than AcCh. PMID- 6278374 TI - Effects of chemicals on delayed matching behavior in pigeons I: acute effects of drugs. AB - Pigeons were trained to peck a red or green center key 15 times to turn it off. After a delay, two side keys, one red and one green were illuminated. Pecks on the side key whose color matched the color that the center key had been produced food. Pecks on the other side key produced a timeout. The effects of various drugs were studied as the delay between extinguishing of the center key light and illumination of side keys was varied from 1 to 8 sec. Pentobarbital and phencyclidine consistently decreased matching, but morphine, d-amphetamine and delta 9-tetrahydrocannabinol had little effect on matching even at doses that increased latency to respond and decreased the rate of responding. Pentobarbital frequently decreased matching at doses below those that increased response latency and decreased response rate, but phencyclidine decreased matching only at doses that increased latencies and decreased rates. The effects of pentobarbital on matching did not depend on the delay duration. PMID- 6278375 TI - Aujeszky's disease: recent advances and current problems. PMID- 6278376 TI - Feline leukaemia virus testing. PMID- 6278377 TI - [In vitro maturation of mammalian oocytes. I. The hormonal regulation of maturation]. AB - A review of the data on in vitro induction of meiotic maturation in cultured follicles and oocytes in mammals. The data are given on cyclic AMP accumulation, steroidogenesis and glycolysis in preovulatory follicles and on the effect of the substances synthesized, as well as the follicular constituents (cumulus and granulosa cells and follicular fluid) on oocyte maturation. Hypotheses concerning the mechanism of gonadotropin - induced mammalian oocyte maturation are discussed. PMID- 6278378 TI - Development of oral HSV-1 infection model in mice. Evaluation of efficacy of 5' amino-5-iodo-2',5'-dideoxyuridine. AB - We developed a new model of oral HSV-1 infection in mice. After oral inoculation, 100 percent of mice developed the clinical lesions at the inoculated area and latent HVS infection in their trigeminal ganglia without mortality. The antiherpetic efficacy of AIdUrd, an agent specifically activated by herpesvirus encoded enzyme, has been evaluated in this animal model. Early topical or systemic treatment of AIdUrd notably reduced the development of clinical lesions and the virus content in the inoculated lips. However, the establishment of latent HSV infection in the sensory ganglia was not influenced by AIdUrd treatment. PMID- 6278379 TI - [Congenital varicella syndrome]. PMID- 6278380 TI - [Myocarditis in the newborn infant caused by Coxsackie B2 enterovirus]. PMID- 6278381 TI - Mixed tumor of the epiglottis: case report. PMID- 6278383 TI - Case report. Hepatocellular carcinoma with lamellar fibrosis: an important histological variant. AB - A case of hepatocellular carcinoma with lamellar fibrosis in a young woman is described. This tumour, in contrast to the usual hepatocellular carcinoma, occurs in young adults, is characterized histologically by large polygonal eosinophilic hepatocytes and abundant fibrous stroma, and has a better prognosis. It has been confused with focal nodular hyperplasia. PMID- 6278382 TI - [Comparative study of light and electron microscopic aspects of myocardial lesions induced in the rat by oils with several amounts of gamma-linolenic acid (author's transl)]. AB - Four different oils : onager oil, mixture of onager oil (20 p. cent) and middle class triglyceride, sunflower oil and soybean oil were given to the rat during three different periods of time : 1) during 3 months after weaning ; 2) during 6 months after weaning ; 3) from the 9th to 15th months. Three types of myocardial lesions were found : interstitial inflammatory reaction, foci of cellular necrosis and fibrosis. The semi-quantitative study of these lesions did not find important differences from one oil to the other, as well as with the controls. However, subgroups onager are among the least altered, particularly as far as necrotic lesions are concerned. Morphometry of mitochondria, despite evident alterations under electron microscope, gave similar results in every groups. PMID- 6278384 TI - [Relationship between the type of bronchial cancer and the presence of cellular infiltrates in and around its foci]. PMID- 6278385 TI - [Adenocarcinoma arising in adenomyosis after estrogen therapy in the menopause]. PMID- 6278386 TI - [A rare case of paraganglioma with intestinal localization]. PMID- 6278387 TI - Lung ossifications in a case of acromegaly. PMID- 6278388 TI - Multicystic nephroblastoma: ultrasound diagnosis (with a pathologic-anatomic commentary). AB - Multicystic nephroblastoma is a cystic variant of the Wilms' tumor. As such, pathologic identification of blastemal elements in multilocular cystic disease is important in the long term care of patients. A case of this unusual Wilms' tumor is described. PMID- 6278389 TI - Antibody response to herpes simplex virus type 1 polypeptides and glycoproteins in primary and recurrent infection. AB - Sequential sera from a patient with primary Herpes Simplex Virus type I (HSV-1) encephalitis and a patient with HSV-1 recurrent oral lesions were collected. Sera were analyzed quantitatively by radioimmunoassay and qualitatively by electrophoresis and autoradiography of immune precipitates to determine the sequence of antibody production to specific radiolabeled HSV-1 polypeptide and glycoprotein antigens. The major antibody response in both primary and recurrent sera was against HSV-1 envelope antigens and the major capsid polypeptide. Sequential sera showed a significant correlation between neutralizing antibody titers and quantitative antibody to HSV-1 glycoproteins. Qualitative electrophoretic analysis of primary infection sera showed sequential appearance of antibodies to increasing numbers of HSV-1 polypeptides by the fourteenth day of infection. A corresponding qualitative antibody response to glycoproteins was not seen. Sequential sera obtained before, during, or after a recurrent lip lesion in another patient showed no significant quantitative or qualitative changes in antibodies to either HSV-1 glycoproteins or polypeptides. PMID- 6278390 TI - Alpha 1-antitrypsin phenotype: transient cathodal shift in serum of infant girl with urinary cytomegalovirus and fatty liver. AB - A 2-month-old white girl had severe liver disease (but without signs of hepatic necrosis, infection or cirrhosis), urinary cytomegalovirus, transient reduction of alpha 1-antitrypsin concentration and transient abnormal alpha 1-antitrypsin phenotype that were not present in her parents. Five serum specimens that were obtained during the 11/2 months of acute phase liver disease indicated, by polyacrylamide gel isoelectric focusing (PAG-IEF), acid starch gel and agarose electrophoresis as well as immunofixation, an unusual alpha 1-antitrypsin phenotype that we labeled delta (delta). It migrated adjacent to Z, i.e., cathodal of Z and Zpratt on PAG-IEF; anodal of Z but cathodal of X, S, Zpratt on starch gel. We labeled the girl's complete phenotype M delta. After clinical recovery, her phenotype was MM and identical to that of her parents. Hepatic electronmicroscopy of the acute phase specimen showed dilated bile canaliculi. We observed the following in hepatocytes: clusters of globular inclusions surrounded by myelin sheets that, to a lesser extent, also appeared in the liver of CMV infected children with phenotype MM; dilated endoplasmic reticulum cisternae that contained floccular material; and marked steatosis. These changes were less severe in the convalescent liver specimen. PMID- 6278392 TI - Biological activities of cystic fibrosis serum. IV. Stimulation of the calcium mediated K+ efflux from rat submandibular gland fragments. AB - Cystic fibrosis (CF) and heterozygote sera stimulate a significant K+ efflux from rat submandibular gland fragments in the presence of 1 mM ouabain. This sensitive parameter can be maximally stimulated by as little as 0.5% CF serum and is inhibited by the calcium channel blocker D600 and EGTA. Specific receptor blockers propranolol, phenoxybenzamine or atropine do not inhibit the CF serum stimulated K+ efflux and agonists do not supramaximally stimulate K+ efflux when added with serum. CF serum-induced K+ efflux did not result in the leakage of lactic dehydrogenase into the bathing media nor did it mimic the action of the calcium ionophore A23187 when added in the presence of D600. In addition, ultrafiltrates of CF serum (less than 10,000 daltons) also stimulated K+ efflux from rat submandibular gland tissue fragments. PMID- 6278391 TI - Pyruvate kinase and phosphofructokinase isozymes in childhood cancers. AB - Pyruvate kinase and phosphofructokinase isozymic expression have been investigated in 62 childhood cancers by electrofocusing and immunoneutralization. Isozymic expression was roughly intermediate between that of the corresponding normal adult and fetal tissue. There were unique features distinguishing malignant from both adult and fetal tissues. In addition, phosphofructokinase and pyruvate kinase appeared to be excellent markers for nervous tissue and skeletal muscle differentiation. In neuroblastomas there was a close relationship between the differentiation level of tumors and their isozyme composition. The same phenomenon was observed in rhabdomyosarcomas and teratomas. PMID- 6278393 TI - Characterization of nuclear 3,5,3'-triiodothyronine receptors in the developing rat lung: effects of hypo- and hyperthyroidism. AB - In this study, we measured the changes in binding characteristics of T3 to its nuclear receptor in lungs of rats from 2 days of age to adulthood. In all ages studied, we found a single class of binding sites with a mean Ka of 1.16 +/- 0.05 x 10(10) M-1. The specificity of the nuclear receptor, as judged by the relative affinities of thyroid hormone analogs, is in general agreement with data previously reported for lung and other organs. A significant decrease in the maximum binding capacity of the lung T3 receptor occurs between 30 and 40 days of age: 0 to 30 days, 0.208 +/- 0.005 pmole T3/mg protein; 40 days to adulthood, 0.111 +/- 0.007 pmole T3/mg protein. Our experiments showed that this reduction is not due to differential recovery of nuclei or efficiency of extraction at various ages. The relative saturation of the nuclear receptor increases from 26% at 2 days of age to 48% in the adult animal. Thyroid status has no significant effect on the nuclear T3 binding capacity of developing rat lung up to 11 days of age. However, hyperthyroidism substantially increased the binding capacity in lungs of adult animals (0.157 +/- 0.009 versus 0.108 +/- 0.006 pmole T3/mg protein), without a corresponding reduction in hypothyroidism (0.116 +/- 0.007 pmole T3/mg protein). These findings suggest that thyroid hormones play a yet unspecified important role in lung physiology during the first month of life in the rat. PMID- 6278394 TI - [Syndromes of developmental anomalies of branchial arches and clefts in the light of cardiological observations]. PMID- 6278395 TI - [Case of viral myocarditis with exudative pericarditis caused by Coxsackie B3 virus]. PMID- 6278396 TI - Ectopic immature renal tissue over the dorsum of lumbar and sacral area in two infants. AB - During reparative surgery for meningomyelocele of the lumbar area, a 7-week-old female infant was found to have a small, well-delineated, subcutaneous, renal blastema. A 3-month-old female infant was found to have immature renal tissue, consisting of glomeruli and tubules, in a soft tissue swelling dorsal to the sacrum. Neither of these patients showed neurologic or renal abnormalities. The second patient has had a disease-free follow-up period of six years. The possible etiology and significance of these benign findings and their relation to the origin of Wilms tumors are discussed. PMID- 6278397 TI - [Characteristics of the cellular and humoral immune response in children with adenovirus infections]. PMID- 6278398 TI - Effects of bilateral amygdala lesions on neophobia and conditioned taste aversion in mice. PMID- 6278399 TI - [Hepatitis B and the prevention of hepatocellular carcinoma]. PMID- 6278400 TI - [Cushing's disease with corticotropic microadenoma and empty sella turcica]. PMID- 6278401 TI - RSITE: a computer program to predict the recognition sequence of a restriction enzyme. AB - A computer program (RSITE) was developed which predicts the recognition sequence of a restriction endonuclease. The sizes of fragments experimentally determined on cleavage of a DNA of known sequence were input. Possible recognition sequences producing fragments of sizes matching those determined empirically were printed out. The program faithfully predicted the specificity of restriction enzymes of known recognition sequence and also determined the recognition sequence of a new restriction enzyme from Haemophilus influenzae GU (HinGU II). PMID- 6278402 TI - Creation of a data base for sequences of ribosomal nucleic acids and detection of conserved restriction endonucleases sites through computerized processing. AB - As part of a project pertaining the organization of ribosomal genes in Kinetoplastidae, we have created a data base for published sequences of ribosomal nucleic acids, with information in Spanish. As a first step in their processing, we have written a computer program which introduces the new feature of determining the length of the fragments produced after single or multiple digestion with any of the known restriction enzymes. With this information we have detected conserved SAU 3A sites: (i) at the 5' end of the 5.8S rRNA and at the 3' end of the small subunit rRNA, both included in similar larger sequences; (ii) in the 5.8S rRNA of vertebrates (a second one), which is not present in lower eukaryotes, showing a clear evolutive divergence; and, (iii) at the 5' terminal of the small subunit rRNA, included in a larger conserved sequence. The possible biological importance of these sequences is discussed. PMID- 6278403 TI - A simple computer program for calculating, modifying and drawing circular restriction maps. AB - An HPL program is described which constructs and draws circular restriction maps given a set of cleavage sites, together with deletion of insertion data if required. This program is of great use in the routine interpretation of restriction patterns for the analysis of recombinant DNA molecules. PMID- 6278404 TI - Computer programs for the analysis and the management of DNA sequences. AB - A program package is described for the management and the analysis of DNA sequence data. The programs - with the exception of a few Fortran routines - are written in the programming language APL. They are best used interactively although batch processing is possible. The package has been in constant use for about 3 years and contains programs for most of the routine problems presently found in a DNA sequencing laboratory. PMID- 6278405 TI - Automatic construction of restriction site maps. AB - A computer program is described which constructs maps of restriction endonuclease cleavage sites in DNA molecules, given only the fragment lengths. The program utilizes fragment length data from single and double restriction enzyme digests to generate maps for linear or circular molecules. The search for a map can be limited to the unknown (insert) region of a recombinant phage or plasmid. Typical restriction maps with four or five enzymes which cut at three to five unknown sites can be calculated in a few minutes. PMID- 6278406 TI - A flexible new computer program for handling DNA sequence data. AB - A compact new computer program for handling nucleic acid sequence data is presented. It consists of a number of different subsets, which may be used according to a given code system. The program is designed for the determination of restriction enzyme and other recognition sites in correlation with translation patterns, and allows tabulation of codon frequencies and protein molecular weights within specified gene boundaries. The program is especially designed for detection of overlapping genes. The language, is FORTRAN and thus the program may be used on small computers; it may also be used without any prior computer experience. Copies are available on request. PMID- 6278407 TI - MAXAMIZE. A DNA sequencing strategy advisor. AB - The MAXAMIZE advisory system determines from user-provided restriction maps an optimal strategy to do nucleotide sequencing by methods involving end-labeled fragments. The maps may be either simple linear restriction maps of fragments or complex circular maps including restriction sites of a vector. The whole system is interactive and is written in the Genetic English language provided by the GENESIS System, a molecular genetics knowledge representation and manipulation package. In addition, MAXAMIZE provides bookkeeping facilities for sequencing and offers advise on how to verify the newly obtained sequence data. PMID- 6278408 TI - Microcomputer programs for DNA sequence analysis. AB - Computer programs are described which allow (a) analysis of DNA sequences to be performed on a laboratory microcomputer or (b) transfer of DNA sequences between a laboratory microcomputer and another computer system, such as a DNA library. The sequence analysis programs are interactive, do not require prior experience with computers and in many other respects resemble programs which have been written for larger computer systems (1-7). The user enters sequence data into a text file, accesses this file with the programs, and is then able to (a) search for restriction enzyme sites or other specified sequences, (b) translate in one or more reading frames in one or both directions in order to find open reading frames, or (c) determine codon usage in the sequence in one or more given reading frames. The results are given in table format and a restriction map is generated. The modem program permits collection of large amounts of data from a sequence library into a permanent file on the microcomputer disc system, or transfer of laboratory data in the reverse direction to a remote computer system. PMID- 6278409 TI - Secondary structure model for the complete simian virus 50 late precursor mRNA. AB - Structures for all sequences containing less than 1790 nucleotides in the 2600 nucleotide late region of the SV40 virus have been computed and saved on magnetic tape. Previously the longest sequence whose secondary structure was calculated in a single computer run contained 950 nucleotides. In the past, analysis of long molecules required numerous repeated, partially overlapping computations on much shorter segments. The structure obtained for the late half of the SV40 is Y shaped with two unequal arms. It has 52 short hairpins. Two long range interactions between nucleotides near 650 and 1350 and between 1450 and 2450 appear to play an important role. The first is within the 16S intron; the second is in the 3' exon. The 5' and 3' ends of the molecule are close to each other and are found in the major elongated stem in the vicinity of the fork. PMID- 6278410 TI - Apple II software for M13 shotgun DNA sequencing. AB - A set of programs is presented for the reconstruction of a DNA sequence from data generated by the M13 shotgun sequencing technique. Once the sequence has been established and stored other programs are used for its analysis. The programs have been written for the Apple II microcomputer. A minimum investment is required for the hardware and the software is easily interchangeable between the growing number of interested researchers. Copies are available in ready to use form. PMID- 6278411 TI - Interactive computer programs in sequence data analysis. AB - We present interactive computer programs for the analysis of nucleic acid sequences. In order to handle these programs, minimum computer experience is sufficient. The nucleotide sequence of the human gamma globin gene complex is used as an example to illustrate the data analysis. PMID- 6278412 TI - A convenient and adaptable package of DNA sequence analysis programs for microcomputers. AB - We describe a package of DNA data handling and analysis programs designed for microcomputers. The package is convenient for immediate use by persons with little or no computer experience, and has been optimized by trial in our group for a year. By typing a single command, the user enters a system which asks questions or gives instructions in English. The system will enter, alter, and manage sequence files or a restriction enzyme library. It generates the reverse complement, translates, calculates codon usage, finds restriction sites, finds homologies with various degrees of mismatch, and graphs amino acid composition or base frequencies. A number of options for data handling and printing can be used to produce figures for publication. The package will be available in ANSI Standard FORTRAN for use with virtually any FORTRAN compiler. PMID- 6278413 TI - Computer programs to analyze DNA and amino acid sequence data. AB - Extensive modifications have been incorporated into many of the computer programs written by Staden (1-4) to make them easier to cope with DNA and amino acid sequence data. These programs can be easily used by persons with minimal knowledge of computers. PMID- 6278414 TI - Computer programs for nucleic acid sequence manipulation. AB - Computer programs are described which help during the collection and analysis of nucleic acid sequence data. They are written in FORTRAN and have been implemented on a PDP 11/60 computer. PMID- 6278415 TI - Is DNA unwound by the cyclic AMP receptor protein? AB - Superhelical pBR 322 derivatives have been relaxed by eukaryotic topoisomerase I in the presence or in the absence of E. coli cyclic AMP receptor protein (CRP) and of cyclic AMP (cAMP). CRP alone, or cAMP alone do not affect the average linking number of the distribution of the relaxed topoisomers. Hence, they do not unwind the template. In the presence of cAMP, CRP induces a small unwinding. The extent of this unwinding is barely modified when the relaxation is carried out on a similar vector plasmid where the CRP binding site of the lac or of the gal operon has been inserted. Under these conditions, we checked that CRP occupies the lactose control site and that upon addition of RNA polymerase, the corresponding promoter is readily activated. These findings are difficult to reconcile with the proposal that activation of these promoters results from the binding of the CRP-cAMP complex to left-handed DNA sequences. PMID- 6278417 TI - A 3' co-terminus of two early herpes simplex virus type 1 mRNAs. AB - A 3' co-terminus of two early herpes simplex virus type 1 mRNAs has been identified using the nuclease -S1 mapping procedure with cloned virus DNA probes. These mRNAs (5.0 kb and 1.2 kb), located within the genome region 0.56-0.60, are unspliced and are transcribed rightwards on the prototype genome orientation. The position of their 3' ends has been located on the virus DNA sequence and lies downstream from the polyadenylation signal 5'-AATAAA-3'. This hexanucleotide sequence also was present in the complementary DNA strand and was shown to be the polyadenylation signal for a leftwards-transcribed late mRNA. The abundance within the cytoplasm of the 5.0 kb and 1.2 kb mRNAs was investigated. Results indicated that these mRNAs were regulated in concert. It is suggested that sequences at the 3' co-terminus may be involved in their regulation. PMID- 6278418 TI - Evidence for ribosomes involved in splicing of yeast mitochondrial transcripts. AB - We have investigated the processing of transcripts of the split gene COB in yeast mitochondrial DNA from cells whose mitochondrial translation was blocked by chloramphenicol for several generations of cell growth. First analysis of transcripts by electrophoresis and RNA/DNA-hybridization clearly showed that cell growth in the presence of CAP leads to an inhibition of processing yielding an increasing amount of splicing intermediates of the COB transcript and decreasing amounts of the 18S mRNA coding for apocytochrome b. This observation is in accordance with the now widely favoured idea that mitochondrial proteins are involved in splicing of COB transcripts and that their reduction should hamper processing and - therefore - lead to an accumulation of pre-mRNAs. However, further information obtained by pulse-labeling of pre-mRNA in vivo in the presence of CAP for various times shows that even 30 minutes after addition of CAP a reduction of the processing rate is obtained. Based on these findings we conclude that maturation of mtRNAs is not only dependent on mitochondrial proteins, but also on a more direct interaction of the translation machinery and RNA processing whose nature is so far unknown. PMID- 6278416 TI - The 5S RNA genes of Schizosaccharomyces pombe. AB - The genomic arrangement and sequences of S. pombe 5S RNA genes are reported here. The 5S gene sequences appear to be dispersed within the genome, and are found independently of other rRNA genes. The sequences of two 5S genes examined show identical coding regions of 119 base pairs but have widely varying flanking sequences. A tRNAAsp gene is found in the 3' flanking region of one of the 5S genes. The tRNAAsp gene is faithfully transcribed in an X. laevis in vitro system, while the 5S genes are not transcribed in this system. The phylogenetic position of S. pombe is examined through comparison of 5S RNA sequences. PMID- 6278419 TI - The genome of Plasmodium falciparum. I: DNA base composition. AB - Some structural properties of the DNA of Plasmodium falciparum were studied thoroughly using several techniques. Its G+C content was found to be extremely low (17-19%), the lowest reported for a living organism. The DNA seems to be composed only of the four major bases as no methylated bases were detected. This DNA had a Tm value of 62.5 degrees C and its denaturation profile showed no marked intramolecular heterogeneity. PMID- 6278420 TI - Simple repeated sequences in human satellite DNA. AB - In an extensive analysis, using a range of restriction endonucleases, HinfI and TaqI were found to differentiate satellites I, II and III & IV. Satellite I is resistant to digestion by TaqI, but is cleaved by HinfI to yield three major fragments of approximate size 770, 850 and 950bp, associated in a single length of DNA. The 770bp fragment contains recognition sites for a number of other enzymes, whereas the 850 and 950bp fragments are "silent" by restriction enzyme analysis. Satellite II is digested by HinfI into a large number of very small (10 80bp) fragments, many of which also contain TaqI sites. A proportion of the HinfI sites in satellite II have the sequence 5'GA(GC)TC. The HinfI digestion products of satellites III and IV form a complete ladder, stretching from 15bp or less to more than 250bp, with adjacent multimers separated by an increment of 5bp. The ladder fragments do not contain TaqI sites and all HinfI sites have the sequence 5'GA(AT)TC. Three fragments from the HinfI ladder of satellite III have been sequenced, and all consist of a tandemly repeated 5bp sequence, 5'TTCCA, with a non-repeated, G+C rich sequence, 9bp in length, at the 3' end. PMID- 6278422 TI - Moloney murine leukemia virus-induced tumors: recombinant proviruses in active chromatin regions. AB - The DNase I sensitivity of chromosomal DNA regions carrying integrated proviral genomes of Moloney (M-MuLV) and AKR Murine Leukemia Virus (AKR-MuLV), and the cellular homologue of the mos-gene (c-mos) of Moloney Sarcoma Virus (MSV) were studied in tumor tissues of leukemic mice. The genetically transmitted sequences of M-MuLV, AKR-MuLV, and the c-mos gene are all in DNase I resistant chromatin conformations in M-MuLV-induced tumors. Each M-MuLV-induced tumor contained at least one somatically acquired integrated recombinant MuLV genome that displayed two main characteristic features of active chromatin: a) a configuration hypersensitive to DNase I, and b) extensive hypomethylation. DNase I hypersensitive sites were mapped at the junction of cellular sequences and the 5' viral large terminal repeat (LTR). Expression of a recombinant MuLV seems therefore to be a necessary feature to maintain the transformed state. PMID- 6278421 TI - Tissue specificity and organisation of CpG methylation in calf satellite DNA I. AB - Examination of bovine satellite DNA I methylation within CpG dinucleotides has been made by restriction analysis. It is shown that variations in the methylation patterns occur between different tissues (brain, liver, thymus and sperm) . Some of the 8 Hpa II sites present per repeat are clearly undermethylated in sperm as compared to other tissues. Methylation is considered therefore, as a highly specific event. It is also shown that there is a spatial specificity in the methylation pattern of the 3 Hha I sites in all tissues. These results are discussed in the light of methylation and satellite DNA functions. PMID- 6278423 TI - Characterization of the DNA duplication-transposition that controls the expression of two genes for variant surface glycoproteins in Trypanosoma brucei. AB - The genome of Trypanosoma brucei carries over a hundred genes coding for different variants of the major surface glycoprotein. Activation of some of these genes is accompanied by a duplication and transposition of the gene (the basic copy) to another region in the genome where it is transcribed. We present here physical maps of the basic and transposition-activated genes for two surface glycoproteins of Trypanosoma brucei, stock 427. In both cases the transposed segment starts 1-2 kb in front of the coding region and ends within the 3' terminal region of the gene. The DNA segments flanking both transposed genes are indistinguishable and share a 6-kb stretch upstream and a 8-kb stretch downstream of the transposed segment not cut by several restriction endonucleases. The 5' borders of the two transposed segments are homologous and contain sequences present in many copies in the genome. A different repeated sequence has previously been found at the 3' edge of the transposed segment. The replicative transposition may, therefore, involve a unidirectional gene conversion initiated by base pairing between the edges of the transposed sequence and a single expression site elsewhere in the genome. PMID- 6278424 TI - On immunoglobulin heavy chain gene switching: two gamma 2b genes are rearranged via switch sequences in MPC-11 cells but only one is expressed. AB - During B lymphocytes differentiation, switches in the expression of heavy chain immunoglobulin constant region (CH) genes occur by a novel DNA recombination mechanism. We have investigated the requirements of the CH gene switch by characterizing two rearranged gamma 2b genes from a gamma 2b producing mouse myeloma (MPC-11). One of the two gamma 2b genes is present in 2-3 copies per cell (gamma 2b strong hybridizer) while the other is present in approximately 1 copy per cell (gamma 2b weak hybridizer). Genomic clones of the gamma 2b strongly hybridizing gene indicate that this is an abortive switch event between the S gamma 3 and S gamma 2b regions. However, clones of the gamma 2b weakly hybridizing gene suggest a functional rearrangement due to the presence of VH, JH and S mu sequences. The switch-recombination sites of these rearranged gamma 2b genes and those of other CH genes show a high degree of preference for the sequence AGGTTG 5' of either the S mu donor site or the appropriate CH S acceptor site. AGGTTG and its analogs are rare in the S mu region, are somewhat prevalent in s alpha and in the case of S mu are found 5' of a tandemly repeated DNA sequence (GAGCT, GGGGT) comprising most of S mu. PMID- 6278425 TI - Organization of highly repetitive satellite DNA of two Cucurbitaceae species (Cucumis melo and Cucumis sativus). AB - The prominent satellites of the Cucurbitaceae Cucumis melo (melon) and Cucumis sativus (cucumber) have been characterized, in actinomycin/CsCl gradients where the satellite sequences can be separated from ribosomal, organelle, and main band DNA the location of the satellites is different indicating a different GC content. The purified satellite of C. melo is cut by HindIII into a repeat unit of 380 bp; AluI digestion gives rise to two bands (about 80 and 220 bp in size). The HindIII repeat unit if cloned into pBR325 exhibits new recognition sites for HpaII leaving two bands with 150 and 80 bp suggesting methylation of the C/CGG cutting site in the uncloned material. The restriction pattern indicates an internal sequence repeat within the 380 bp HindIII fragment. The C. sativus satellite is cut by AluI to a repeat unit of 180 bp showing no other recognition site for the restriction enzymes tested so far. About 10% sequence homology has been determined between the C. melo and C. sativus satellites by cross hybridization studies. A high methylation degree of cytosines has been measured for both satellites and the ribosomal DNA of C. sativus (about 30%). No transcription products of the C. melo satellite were found during seedling development. PMID- 6278427 TI - Avian erythrocyte chromatin degradation: the progressive exposure of the dinucleosomal repeat by bovine-pancreatic-DNAase-I-armed probes and free DNAase I. AB - Bulky probes composed of glutaraldehyde-cross-linked-complexes of ferritin or bovine serum albumen armed with bovine pancreatic DNAase-I have been used to generate the typical dinucleosome-based series of native or denatured DNA fragments from chicken erythrocytes. Double stranded DNA fragments have shown that the 2N series extends to at least 10N without any clearly defined end-point. The susceptible nucleosomes were observed to be nucleolysed by two major, different methods of attack that generated multiple peaks in the dinucleosomal size class and some characteristics of the digestion patterns have been taken as indications of the presence of another level of structure above that of the alternating structure. The chromatin structure becomes resistant to the probes when the average fragment length falls to approximately 4N but this 'core' is not a single integral multiple of nucleosomes. PMID- 6278426 TI - Secondary structure of Bombyx mori and Dictyostelium discoideum 5S rRNA from S1 nuclease and cobra venom ribonuclease susceptibility, and computer assisted analysis. AB - The 5S rRNAs from Bombyx mori and Dictyostelium discoideum were end-labeled with [32-P] at either the 5' or 3' end and sequenced using enzymatic digestion. The secondary structure of these molecules was studied using the single-strand specific S1 nuclease and the base-pair specific cobra venom ribonuclease. Computer analysis of these results was performed and was used to generate a consensus secondary structure for each molecule. A comparison of these results with those of other workers is presented. PMID- 6278428 TI - The primary structures of two leghemoglobin genes from soybean. AB - We present the complete nucleotide sequences of two leghemoglobin genes isolated from soybean DNA. Both genes contain three intervening sequences which interrupt the two coding sequences in identical positions. The 5' and 3' flanking sequences in both genes contain conserved sequences similar to those found in corresponding positions in other eukaryotic genes. Thus, the general DNA sequence organization of these plant genes is similar to that of other eukaryotic genes. PMID- 6278429 TI - Reduced-stringency DNA reassociation: sequence specific duplex formation. AB - Reduced-stringency DNA reassociation conditions allow low stability duplexes to be detected in prokaryotic, plant, fish, avian, mammalian, and primate genomes. Highly diverged families of sequences can be detected in avian, mouse, and human unique sequence dNAs. Such a family has been described among twelve species of birds; based on species specific melting profiles and fractionation of sequences belonging to this family, it was concluded that permissive reassociation conditions did not artifactually produce low stability structures (1). We report S1 nuclease and optical melting experiments, and further fractionation of the diverged family to confirm sequence specific DNA reassociation at 50 degrees in 0.5 M phosphate buffer. PMID- 6278431 TI - The sequence of the nucleoprotein gene of human influenza A virus, strain A/NT/60/68. AB - The nucleotide sequence of the nucleoprotein gene of influenza A/NT/60/68 was established after using improved cloning methods to obtain full length cDNA clones in pBr322. The gene is 1565 residues long and codes for a basic protein of 498 amino acids. There are only 30 amino acid differences between it and the homologous sequence in A/PR/8/35, all occurring as point mutations. Assuming a common lineage, the evolutionary rate of divergence of the two strains is 0.18% amino acid per year. This confirms there is a slow but significant rate of evolution. PMID- 6278430 TI - Uncoupling of SV40 tsA replicon activation from DNA chain elongation by temperature shifts and aphidicolin arrest. AB - To synchronize SV40 replicons, simian cells infected with a tsA mutant were restricted at 40 degrees, to complete ongoing replication and returned to 32 degrees, to activate new replicons in the presence of the DNA chain elongation inhibitor aphidicolin. Upon further incubation at 40 degrees without the drug, 3H dT was incorporated into SV40 FI DNA, almost to the extent seen with cells recovered in the absence of the drug. To determine whether DNA synthesis would begin from the origin, following the temperature-shifts-aphidicolin regimen, chains subsequently pulse-labeled with (alpha-32p)dGTP in isolated nuclei were analyzed for size distribution and genomic location. These chains reached up to 300-400 nucleotides in size, unlike the control which featured comparable amounts of label in long chains and Okazaki pieces. The nascent DNA of the drug-treated system could be chased into longer chains, indicating that it was a replicative intermediate; and it hybridized preferentially to an origin proximal fragment of AtuI- restricted SV40 DNA, demonstrating partial replicon synchronization. The data prove that T-antigen activates the SV40 replicon independent of DNA chain elongation and suggest means to study the mechanism of DNA chain priming at the origin. PMID- 6278432 TI - Comparison of nucleotide sequences of mRNAs belonging to the mouse H-2 multigene family. AB - The complete nucleotide sequences of three cDNAs coding for the C-terminal part of mouse histocompatibility (H-2) antigens, and for the 3' non coding regions of these clones have been determined. Comparison of the sequence indicates a large homology throughout the coding and non-coding regions and suggests the existence of a genetic mechanism which homogenizes nucleotide sequences among genes of the H-2 multigene family. PMID- 6278433 TI - A novel species of double stranded RNA in mitochondria of Saccharomyces cerevisiae. AB - A double stranded RNA species has been detected in guanidine hydrochloride extracts of mitochondria from respiratory competent cells of Saccharomyces cerevisiae. This novel mitochondrial RNA, termed mtdsRNA, has been purified in a Cs2SO4 density gradient where it bands at a density of 1.58 g/ml. The mtdsRNA runs as a single slow moving band on agarose gels. Its double stranded RNA character was evidenced by its sensitivity to digestion by RNase III, but not by RNase H, or DNase I. Moreover the mtdsRNA hybridized to each separated strand of a petite mtDNA. It is concluded that mtdsRNA contains long transcripts derived from most regions of yeast mtDNA, because 1) its weight-average length as determined by electron microscopy was 4.5 micrometer (about 14 kb, or 20% of the wild type mtDNA genome), and 2) it hybridized to each of a series of eight petite mtDNA probes carrying sequences derived from widely different segments of mtDNA. It is proposed that prolonged transcription of both strands of yeast mtDNA can occur and that mtdsRNA arises from hybridization of these long complementary transcripts. PMID- 6278434 TI - Isolation and structure of a yeast initiator tRNAmet gene. AB - Sixteen bacterial clones containing yeast initiator tRNAmet genes have been isolated. The size of the BamHI fragments encoding these genes ranges from 4,000 to 23,000 base pairs. The nucleotide sequence of one member of this group has been determined. It has no intervening sequences. PMID- 6278435 TI - Study on conformational states of Escherichia coli tRNAPhe in solution by a modulation-free ESR-spectrometer. AB - A modulation free Electron Spin Resonance spectrometer was used for the registration of spectral absorption lines of a spin-labeled Escherichia coli phenylalanine tRNA in solution with low (less than 0.1%) line shape distortion. The analysis of line shape of two different spin-labels introduced into position 8 revealed that phenylalanine tRNA in solution exists as a mixture of two conformers, the equilibria between conformers being dependent on pH, concentration of magnesium and functional state of tRNA (deacylated, aminoacylated or peptidylated). There are no overall structural rearrangements upon aminoacylation or peptidylation of tRNA. The observed small changes of spectral line shape can be assigned to shifts in conformational equilibria. PMID- 6278436 TI - analysis of genomic rearrangements associated with two variable antigen genes of Trypanosoma brucei. AB - Some variable surface glycoprotein (VSG) genes of Trypanosoma brucei undergo duplication and transposition when they are expressed. We report here the cloning of cDNAs coding for two VSGs from the ILtar 1 repertoire. Analysis of the genomes of trypanosomes expressing these and other antigens shows that there is no additional copy of the sequences coding for eight VSG in expressing clones of trypanosomes, and reveals rearrangements analogous to those previously described for the gene for another VSG from this antigen repertoire. The data indicate that duplication does not accompany the expression of these VSG genes. Transposition to a specific expression site cannot be excluded, but would have to involve either a much larger segment of DNA, or movement to a region of much greater homology with the previous flanking sequences, than is observed for VSG genes that are duplicated when expressed. It is reasoned that the control of expression by coupled duplication and transposition is not sufficient to account for the selection of a single VSG gene for expression. PMID- 6278437 TI - Oligonucleotide-directed mutagenesis of gene IX of bacteriophage M13. AB - The synthetic oligodeoxyribonucleotide pCGAAAGACTACAC has been applied as a site specific mutagen to introduce a T leads to G transversion mutation at nucleotide position 1223 of the M13 DNA sequence. The in vitro-induced conversion of a TAT codon into a TAG at this position resulted in gene IX mutants with an amber mutant character thereby confirming that this reading frame defines a gene of an essential phage protein. The gene IX amber mutants obtained grew well on SuI (Ser) and SuIII (Tyr) suppressing strains but could not be propagated on SuII (Gln) and SuVI (Leu) strains. Complementation studies show that amber mutants in genes V and VII exert a polar effect on gene IX expression suggesting that these three contiguous genes form an operon. In addition, we demonstrate the in vitro synthesis of gene IX-protein in a coupled transcription-translation system. PMID- 6278439 TI - Overlapping genes at the DNA primase locus of the large plasmid ColI. AB - The sog gene of the large plasmid ColIdrd-1 has previously been shown to encode a DNA primase and a smaller antigenically related polypeptide. Genesis of these two products has been examined using Sog+ recombinant plasmids. Effects of amber mutations, isolated after in vitro mutagenesis, and deletions into or within sog suggest that the smaller polypeptide is a separate translation product which is encoded by DNA specifying the C-terminal region of the larger protein. Under control of the lac promotor, synthesis of both polypeptides is reduced when transcription is repressed. These findings imply that transcription of sog yields a single transcript which is translated from two initiation sites. PMID- 6278438 TI - Discriminatory function of ribonuclease H in the selective initiation of plasmid DNA replication. AB - The initiation stage of ColE1-type plasmid replication was reconstituted with purified protein fractions from Escherichia coli. The reconstituted system included DNA polymerase I, DNA ligase, RNA polymerase, DNA gyrase, and a discriminating activity copurifying with RNAase H (but free of RNAase III). Initiation of DNA synthesis in the absence of RNAase H did not occur at the normal replication origin and was non-selective with respect to the plasmid template. In the presence of RNAase H the system was selective for ColE1-type plasmids and could not accept the DNA of non-amplifiable plasmids. Electron microscopic analysis of the reaction product formed under discriminatory conditions indicated that origin usage and directionally of ColE1, RSF1030, and CloDF13 replication were consistent with the normal replication pattern of these plasmids. It is proposed that the initiation of ColE1-type replication depends on the formation of an extensive secondary structure in the origin primer RNA that prevents its degradation by RNAase H. PMID- 6278440 TI - A lac promoter with a changed distance between -10 and -35 regions. AB - A lac promotor mutant was constructed by filling in the protruding ends of the HpaII site located within the lac promotor. The mutation, named M42, is a two base pair insertion that changes the distance between the -10 and -35 regions from 18 to 20 residues. The activity of the mutant promotor measured in vivo is 15% of the wild-type promotor. The M42 promotor is sensitive to the catabolite repression in the manner similar to that of the wild type. Sequences of several deletions within the lac promotor are also given. PMID- 6278441 TI - Modification profiles of bacterial genomes. AB - DNAs were prepared from twenty-six bacterial species and digested with a variety of restriction endonucleases to determine what modifications the DNAs carry. Several general conclusions could be made: 1) First, in no instance was the DNA of a restriction enzyme. 2) The specificity of the DNA modification was the same as that of its restriction counterpart; there were no cases of the DNAs being modified against a less specific class of restriction enzymes. 3) In most (but not all) cases, the resistance of a bacterium's DNA to its own restriction enzyme could be generalized to include resistance to all other restriction enzymes with the same specificity (isoschizomers). 4) DNA modified within the central tetramer of a recognition sequence is usually protected against cleavage by all related hexameric enzymes possessing that central tetramer. Only three families of DNA presented in this study disobey this rule. 5) Finally, a significant number of cases emerge where bacterial DNA carries a modification but no corresponding restriction endonuclease activity. PMID- 6278442 TI - Structural analyses of E. coli 5S RNA fragments, their associates and complexes with proteins L18 and L25. AB - The structure of Escherichia coli 5S RNA fragments 1-41 and 42-120 has been studied by the read-off gel sequencing technique using S1 nuclease and cobra venom RNase as probes. Comparison of the digestion patterns with those of reassociated and intact 5S RNA suggests that the structure of both fragments is very similar to that of the corresponding regions in the intact molecule. Six different fragments obtained by partial digestion with T1 RNase and S1 nuclease have been used for reconstitution of 5S RNA, its certain structural regions and complexes with ribosomal proteins L18 and L25 recognizes the double-helix consisting of nucleotides 79-97 (i.e. prokaryotic stem), whereas a loop-region around position 40 (possible positions 39-47) is involved in the interaction with protein L18. PMID- 6278443 TI - DNA sequence of an immediate-early gene (IEmRNA-5) of herpes simplex virus type I. AB - We describe a 2560 base pair herpes simplex virus type 1 (HSV-1) DNA sequence containing the entire immediate-early mRNA-5 (IEmRNA-5) gene. The 3' and 5' termini of IEmRNA-5 were mapped within this DNA sequence by single-strand specific endonuclease protection experiments. The IEmRNA-5 gene contains DNA sequences from both the unique (Us) and reiterated (TRs/IRs) regions of the HSV-1 DNA short component and is interrupted by a single intron mapping in TRs/IRs. A search of the transcribed DNA sequence revealed no initiator codon within TRs/IRs. The first ATG was located 6 bases into Us sequences and this reading frame (316 codons) was also observed in the 3' transcribed region. The oligonucleotide sequences adjacent to the IEmRNA-5 termini are discussed in relation to those of the HSV-1 thymidine kinase gene and other genes transcribed by RNA polymerase II. PMID- 6278444 TI - Preferential site-dependent cleavage by restriction endonuclease PstI. AB - The four identical recognition sites for the restriction endonuclease PstI in purified plasmid pSM1 DNA I are cleaved at markedly different rates. The order and relative frequencies of cleavage at these four PstI sites have been determined from the order of appearance of partial cleavage products and from an analysis of production of specific unit length linear molecules. The same pattern of preferential cleavage is also found when linear, nicked circular, or relaxed closed circular forms of the same plasmid DNa are used as substrates for PstI. Inspection of the nucleotide sequences immediately adjoining each of the PstI sites suggests that the presence of adjacent runs of G-C base pairs confers significant resistance to cleavage. PMID- 6278445 TI - DNase I hypersensitive sites of the chromatin for Drosophila melanogaster ribosomal protein 49 gene. AB - By using an indirect end-labelling technique for mapping, five DNase I hypersensitive sites have been located in Drosophila melanogaster chromatin at the 5'-end of the gene coding for ribosomal protein 49. These sites typically span about 100-600 base pairs and are approximately the length of a nucleosome apart (center to center distance ca 245 bp). This is the first analysis of the chromatin structure of a constitutive house-keeping gene. The results support the hypothesis that the presence of such a DNase 1 hypersensitive site in chromatin is necessary for transcription in vivo. The presence of such sites may reflect some local changes in the conformation of the chromatin in the presumptive regulatory region. PMID- 6278446 TI - In vitro transcription of a cloned mouse ribosomal RNA gene. AB - An in vitro transcription system which utilizes cloned mouse ribosomal RNA gene (rDNA) fragments and a mouse cell extract has been developed. RNA polymerases I is apparently responsible for this transcription as evidenced by the complete resistance to a high concentration (200 micrograms/ml) of alpha-amanitin. Run-off products obtained with three different truncated rDNA fragments indicated that RNA was transcribed from a unique site of rDNA. The S1 nuclease protection mapping of the in vitro product and of in vivo 45S RNA confirmed this site, indicating that, in this in vitro system, transcription of rDNA started from the same site as in vivo. This site is located at several hundred nucleotides upstream from the putative initiation site reported by us (1) and by others (2). Some sequence homology surrounding this region was noted among mouse, Xenopus laevis and Drosophila melanogaster. The data also suggest that some processing of the primary transcript occurs in this in vitro system. PMID- 6278447 TI - Purification and characterization of two new modification methylases: MClaI from Caryophanon latum L and MTaqI from Thermus aquaticus YTI. AB - A method for detecting Type II modification methylases and determining their methylation site by assaying the ability of methylated DNA to be cleaved by heterologous restriction enzymes is described and applied to the isolation of the restriction modification methylases from Thermus thermophilus HB8, Thermus aquaticus YTI and Caryophanon latum L. M.TaqI is shown to have a methylation specificity identical to M.ThI (TCGmeA). M.ClaI methylates at adenine and protects a subset of TthI sites indicating that it methylates the sequence ATCGmeAT. Methylation by M.ThI also protects against cleavage by SalI, XhoI and at some HindII, AccI and MboI sites. PMID- 6278448 TI - Separation of closed circular DNA from linear DNA by electrophoresis in two dimensions in agarose gels. AB - A method that provides an easy, rapid, and reproducible way for separating closed circular DNA species from linear DNA and nicked circles is described. The method is based on the difference in mobility of form I (supercoiled) DNA and form II (nicked circles), and the differential mobility of relaxed circular DNA in the presence and absence of ethidium bromide (EtdBr). It can be used for detection or for purification of plasmid, episomal, or viral DNA from the bulk of cellular DNA, or from other DNA mixtures. PMID- 6278449 TI - Restriction mapping of a new deletion responsible for G gamma (delta beta)o thalassemia. AB - DNA from individuals heterozygous for (G)gamma(deltabeta)(o) thalassaemia has been studied by restriction endonuclease analysis. The results reveal a new molecular defect associated with this condition. A total of three defects is now responsible for the one single phenotype, thereby emphasising the complex relationship between genotype and phenotype among the disorders of beta-like globin synthesis in man. PMID- 6278450 TI - Nucleotide sequence at the end of the gene for the RNA polymerase beta' subunit (rpoC). AB - We have determined the DNA sequence surrounding the transcription terminator following rpoC, the gene that codes for the beta' subunit of RNA polymerase in E. coli K12. The 2044 bp sequence obtained contains the distal 335 codons of rpoC followed by a 212 bp non-coding region and a second open reading frame (ORFa) of 179 codons. The final 181 nucleotides of the sequence form the 5' end of a third open reading frame (ORFb). The in vivo 3' end of the rpoC mRNA was located by analysis of RNA/DNA hybrids cleaved with nuclease S1 (S1 mapping). These results indicated that the major transcription termination of the rplJL-rpoBC transcription unit occurs a short distance past the translation stop codon for rpoC. Four regions of symmetry, suggesting secondary structure in the mRNA, were found in the DNA sequence near the rpoC translation termination codon. The last of these hairpin structures is similar to other rho-independent transcription terminators and its 3' end coincides with the end of the rpoC mRNA as predicted by S1-mapping. Inspection of the open reading frames indicates that rpoC uses a high percentage of codons that are recognized by the major tRNA species of E. coli while ORFa and ORFb contain many codons recognized by minor tRNA species. ORFa specifies a very basic peptide. PMID- 6278451 TI - Conservation of sequence arrangement among higher plant chloroplast DNAs: molecular cross hybridization among the Solanaceae and between Nicotiana and Spinacia. AB - Isolated, nick-translated Pvu II fragments of Nicotiana tabacum chloroplast DNA produce specific intra- and intergeneric hybridization signals with chloroplast DNA digests from several representatives of the Solanaceae. These data, along with similarities in restriction enzyme patterns, permit construction of physical maps for Nicotiana line 92 (a cytoplasmic substitution line), Atropa belladonna and Petunia parodii. Plastid-DNA map differences among the Solanaceae are shown to result from single base-pair substitutions as well as local deletions or insertions. Several of these differences of Nicotiana tabacum chloroplast DNA fragments to a chloroplast DNA digest of Spinacia oleracea defines a sequential arrangement of fragments for spinach DNA which is very similar to its published physical map. This is achieved although chloroplast-DNA restriction enzyme patterns from the two organisms are grossly dissimilar. Alignment differences which have been revealed involve the edges of the inverted repeat region where certain single copy stretches in tobacco have been duplicated in spinach. PMID- 6278452 TI - A complex repeated DNA sequence within the Drosophila transposable element copia. AB - A 320 nucleotide repeated DNA sequence within the copia coding element of Drosophila melanogaster has been identified and characterized. This sequence has been localized by DNA-DNA hybridization and electron microscopic analysis of heteroduplexes to the approximate middle of the 5 kb copia coding region. The primary sequence of this repeated DNA has been determined. The sequence is composed of three related subunits, 35-37 nucleotides in length (A, B and C). This 105 nucleotide higher order repeat has apparently been duplicated twice to yield a complex repeated sequence, ABCA'B'C'A"B"C", which exhibits divergence among the individual subunits. This sequence is AT rich, as are the direct terminal repeats which flank the copia coding region, but does not contain any apparent homology with the terminal repeats. This repeated sequence contains three presumptive polyadenylation signals and two 25 nucleotide, imperfectly matched, inverted repeat sequences adjacent to two of the polyadenylation sequences. PMID- 6278453 TI - Two cloned beta thalassemia genes are associated with amber mutations at codon 39. AB - Two beta globin genes from patients with the beta(+) thalassemia phenotype have been cloned and sequenced. A single nucleotide change from CAG to TAG (an amber mutation) at codon 39 is the only difference from normal in both genes analyzed. The results are consistent with the assumption that both patients are doubly heterozygous for beta(+) and beta degrees thalassemia, and that we have isolated and analyzed the beta degrees thalassemia gene. PMID- 6278454 TI - Nuclear Overhauser effect study and assignment of D stem and reverse-Hoogsteen base pair proton resonances in yeast tRNAAsp. AB - Nuclear Overhauser effects (NOEs) in yeast tRNAAsp were found for all four GU and G psi base pairs. NOEs of both reverse-Hoogsteen pairs were identified by comparison with a purine C8 deuterated sample. Several NOEs involving these resonances were also found which are clearly between single protons on adjacent base pairs. These interbase NOEs, combined with the assumption of reasonable similarity between the structure of yeast tRNAAsp and that of yeast tRNAPhe, lead to unambiguous assignment of many resonances including all the ring NH and C2 protons in the D stem. The stability of the stem at 28 degrees C, as recently deduced by Moras et al (Nature 288 669-674), from x-ray diffraction is confirmed. Assignments of the ring NH resonances of T54-A58 and of a G psi pair are made for the first time. PMID- 6278455 TI - Myocardial aneurysm in association with disseminated cytomegalovirus infection. AB - An infant with disseminated cytomegalovirus infection and apical aneurysm of the left ventricle died. At autopsy the coronary arteries were anatomically normal, but there was occlusion of the left anterior descending artery with an inflammatory lesion and corresponding organized thrombus. It seemed likely that cytomegalovirus infection acquired in utero may have induced an endothelial lesion, leading to thrombosis, occlusion, apical myocardial infarction, and eventual aneurysm formation. PMID- 6278457 TI - ACTH secreting pituitary adenoma in an infant of 18 months. Immunohistochemical, electron-microscopic, and in-vitro studies. AB - An 18-month old infant with Cushing's disease due to an ACTH producing pituitary tumor is presented. The case showed typical clinical and morphological sings of hypercortisolism. The infant died of pulmonary thromboembolism after transsphenoidal partial adenomectomy. The adrenals were diffusely hyperplastic. The pituitary adenoma was classified as an undifferentiated mucoid cell adenoma with sparse granulation by light microscopy. Immunoenzymatic studies demonstrated ACTH not only in granulated adenoma cells. Ultrastructurally the cells were only differentiated as typical ACTH cells or so-called follicular cells in small areas. Most of them were undifferentiated, showing pleomorphism of the relatively sparse organelles. In-vitro experiments using suspensions of adenoma cells showed a distinct enhancement of ACTH secretion after arginine-vasopressin and a further decrease ultrastructurally in the number of secretory granules. No effect of ACTH levels and no alterations of the ultrastructure were observed after cortisol. The case is representative of typical hypothalamic-hypophyseal Cushing's disease with an undifferentiated pituitary adenoma secreting ACTH in part autonomously. This constellation of Cushing's syndrome is extremely rare at the age of one year. Our case is the second one reported in the literature. PMID- 6278456 TI - [Etiology and treatment of atopic dermatitis in the light of new data]. PMID- 6278458 TI - Malignant fibrous histiocytoma of bone. A clinico-pathological and electronmicroscopical study. AB - The clinicopathological and ultrastructural features of seven Malignant Fibrous Histiocytomas reported to the Spanish Registry of Bone Tumors were studied. Two patients were females (20 and 60 years old) and the other five males (15, 37, 67, 38 and 17 years old). The tumors were located in the lower femur (3 cases), upper tibia, upper humerus, rib and iliac bone. The past medical history revealed previous fracture in one patient and previous radiation of the affected bone in another; in a third patient the tumor associated to a bone infarct. Microscopically all tumors presented a storiform pattern composed of neoplastic fibroblasts, histiocytes, and malignant multinucleated giant cells. There was no evidence of osteoid or bone tissue formation by the tumor cells. Five cases were studied with electronmicroscopy; the tumor cells had features of fibroblasts, myofibroblasts, histiocytes and multinucleated histiocytes. Primary and secondary lysosomes, along with lipid vacuoles were common findings in the tumor cells. The value of the electron-microscopy in the differential diagnosis of this tumor is emphasized. PMID- 6278460 TI - New quinoline derivatives as amoebicidal and cysticidal agents. PMID- 6278459 TI - The primary tissue culture of human adrenocortical Conn's adenomata. I. The synergistic stimulation of adenomatous cell growth by purine cyclic nucleotides and by ACTH1-24 and angiotensin II. AB - Primary cultures of dissociated parenchymal cells from Conn's adenomata causing primary hyperaldosteronism were successfully set up by a method previously used with normal adult human and rat adrenocortical tissue. In such cultures the adenomatous cells largely prevailed (making up 87% of the whole cell population), could survive for lengthy terms (at least up to 30 days), and were endowed with a spontaneous, discrete capability to proliferate. The de novo RNA- and DNA synthetic and mitotic activities of Conn's cells were markedly stimulated in cultures exposed between 16 and 21 to daily doses of exogenous cyclic AMP, either alone or in equimolar association with cyclic GMP. A significantly weaker, though still prominent enhancement of adenomatous cell growth was elicited also by daily administrations of an equimolar mixture of ACTH1-24 and angiotensin II. In contrast, little stimulation or inhibition of growth or no effect at all could be observed when cyclic GMP, ACTH1-24, and angiotensin II were respectively administered, each by itself. PMID- 6278461 TI - [What is your diagnosis? Arrhenoblastoma]. PMID- 6278462 TI - [Use of glucocorticoid preparations in congenital adrenocortical dysfunction]. AB - Circadian changes in the corticosteroid, testosterone, and ACTH content in the blood plasma were examined to monitor the suppressant effect of glucocorticoids on adrenocortical and hypophyseal functions during administering glucocorticoid drugs to patients with congenital adrenocortical dysfunction. The daily dose of glucocorticoids was given in two intakes according to the two schemes: scheme 1 included the drug intake in the morning and in the afternoon, scheme 2 in the morning and in the evening. The effect of glucocorticoid therapy on the pattern of the circadian rhythm and the absolute corticosteroid and testosterone content was shown to depend on the time of the drug intake. The treatment according to scheme 2 is preferable: the absolute testosterone content was reducing to normal during the whole day. Meanwhile the normal circadian rhythm of the adrenal cortex and hypophysis was preserved. PMID- 6278463 TI - [Catecholamine regulation of the liver and kidney ATPase activity in mice]. AB - Noradrenaline (NA) and isopropyladrenaline in a dose of 11 microM/kg were shown to activate in vivo Na+, K+-ATPase and Mg2+-ATPase in the mouse renal microsomal fraction. NA (10(-6) M) increased the activity of Na+, K+-ATPase in vitro and did not affect the activity of Mg2+-ATPase in the hepatic and renal microsomal fractions. NA did not significantly affect in vivo the mitochondrial ATPases activity in the presence of various stimulators (Mg, dinitrophenol, bicarbonate) and inhibitors (oligomycin, bicyclohexyl carbodiimide) in the mouse liver and kidneys. cAMP (10(-6) M) activated Na+, K+-ATPase in vitro reduced the activity of Mg2+-ATPase in the microsomal fraction, and did not change the activity of the mitochondrial ATPases in the mouse liver and kidneys. The following scheme of the mechanism of catecholamine (CA) effect on Na+, K+-ATPase is suggested: CA leads to beta-adrenoreceptors leads to adenylate cyclase leads to cAMP leads to protein kinase leads to Na+, K+-ATPase. The mechanism of Mg2+-ATPase regulation is discussed. PMID- 6278464 TI - Incorporation into DNA of the base analog 2-aminopurine by the Epstein-Barr virus induced DNA polymerase in vivo and in vitro. AB - The Epstein-Barr virus (EBV)-induced intracellular DNA polymerase was assayed in vitro for the ability to utilize the mutagenic nucleotide analog 2-aminopurine deoxyribose triphosphate (d2apTP), incorporating it as the corresponding monophosphate into DNA or poly[d)(A-T)] template. Bacteriophage T4, lymphocyte alpha, and the EBV particle-associated DNA polymerases were assayed simultaneously for direct comparison. Unlike these three polymerases, which were capable of distinguishing between d2apTP and dATP with a strong preference for the latter, the EBV-induced DNA polymerase only weakly distinguished between dATP and d2apTP and incorporated substantial amounts of d2apTP into template. Detergent-treated lymphocyte nuclei undergoing a high level of EBV DNA synthesis were shown to incorporate the 2-aminopurine analog of dATP into viral DNA. The relative inability of the EBV-induced DNA polymerase to distinguish between the two purine nucleotides reported here is consistent with previous reports on the ready incorporation of other nucleotide analogs into DNA polymerases induced by other herpesviruses. Because most antiherpes agents currently in use or under study are nucleotide analogs, the viral mutagenic properties of these drugs should be examined. PMID- 6278465 TI - Mutual interaction between adjacent dG . dC actinomycin binding sites and dA . dT netropsin binding sites on the self-complementary d(C-G-C-G-A-A-T-T-C-G-C-G) duplex in solution. AB - The Watson-Crick imino protons, the backbone phosphodiester resonances, and the antibiotic exchangeable protons have been used as markers to monitor the separate and simultaneous binding of actinomycin and netropsin to the d(C-G-C-G-A-A-T-T-C G-C-G) self-complementary duplex in aqueous solution. We demonstrate that intercalation of actinomycin at dG(3'-5')dC sites at either end of the duplex results in a conformational perturbation at the dA . dT tetranucleotide core of the dodecanucleotide duplex. Parallel studies of the groove binding of netropsin at dA . dT sites in the interior of the duplex reveal a conformational perturbation which extends to adjacent dG . dC base pairs in the dodecanucleotide duplex. The NMR markers demonstrate that the d(C-G-C-G-A-A-T-T-C-G-C-G) duplex can accommodate actinomycin and netropsin simultaneously at adjacent dG . dC and dA . dT tetranucleotide blocks along its length with some mutual interaction between neighboring antibiotic binding sites. PMID- 6278466 TI - beta-Endorphin: demonstration of a tertiary structure in aqueous solution. AB - Difference spectra generated during thermolysin digestion of camel beta-endorphin at pH 8.2 or at pH 6.5 indicate rapid blue shifting of the near-UV absorption bands of the NH2-terminal tyrosine. A similar spectral change is not observed for the NH2-terminal tyrosine in [Met]enkephalin when it is digested under similar conditions. These results suggest that enzymatic digestion destroys or alters some structural interaction between the NH2-terminal tyrosyl residue of the endorphin and a residue(s) within the COOH-terminal segment of the molecule. Peptide mapping of the digest as a function of time suggests that cleavage of the bond linking the alanine-21 and isoleucine-22 residues produces most of the observed effect. These data provide evidence for the existence of a tertiary structure for beta-endorphin in aqueous solutions. PMID- 6278467 TI - Synthetic glycopeptide substrates for receptor-mediated endocytosis by macrophages. AB - Mammalian macrophages contain a transport system that binds and internalizes glycoproteins with exposed mannose residues. This system and analogous systems on other types of cells require substrates to bear multiple nonreducing terminal residues of the appropriate sugar for effective uptake. Small multivalent synthetic glycopeptides with mannose residues covalently linked through a spacer arm to the alpha- and epsilon-amino groups of lysine, dilysine, and trilysine are competitive inhibitors of rat alveolar macrophage uptake of the neoglycoprotein mannosyl-bovine serum albumin with inhibition constants in the microM range. Various compounds could be covalently attached to the alpha-carboxyl group of these glycopeptides with substantial retention of inhibitory potency. This uptake system does not recognize galactose residues, and the galactosyl analog of an inhibitory mannosylpeptide did not inhibit uptake of mannosyl-bovine serum albumin. The trimannosyldilysine ligand is not only an inhibitor but also a substrate for specific uptake by macrophage, as shown with an 125I-labeled derivative. Macrophages bound 6.4 x 10(5) molecules per cell at 0 degrees C with a dissociation constant of 2 microM. At 21 degrees C the cells could internalize the labeled conjugate with an apparent Michaelis constant of 6 microM and a maximal velocity of 1.7 x 10(5) molecules per min per cel. The dissociation constant and Michaelis constant are similar to the inhibition constant of 9 microM determined at 21 degrees C for inhibition by this conjugate at mannosyl bovine serum albumin uptake. These synthetic substrates may be useful in targeting pharmacologic agents to macrophages, and analogous compounds may target such agents to other types of cell. PMID- 6278468 TI - [3H]Methotrexate as a ligand for the folate receptor of Dictyostelium discoideum. AB - Studies of the folate chemotactic receptor of vegetative Dictyostelium discoideum cells have been hampered by the presence of the degradative enzyme folate deaminase. The diaminopterin compounds aminopterin and methotrexate (MTX) are chemoattractants but are not attacked by the deaminase. [3',5',7,9 3H]methotrexate ([3H]MTX) is a nondegraded radioligand for the folate receptor. Binding to the receptor is rapid, reaching steady state in less than one min, and reversible in less than 15 s by an excess of unlabeled MTX. A single class of binding sites is found with a Kd of 2 x 10(-8) M, which correlates well with the concentration dependence of chemotaxis. Folate, aminopterin, and MTX all compete for [3H]MTX binding, whereas pterin, p-aminobenzoate, and nucleotides do not. Analysis of the receptor during differentiation indicates a decrease in site number by a factor of 3 with no change in affinity during the first 7 hr. During this time, the directional response (chemotaxis) to MTX and folate is lost, but a nondirectional stimulation of motility rate (chemokinesis) is retained. The response to cyclic AMP displays reciprocal behavior, first appearing as a chemokinetic response and then as a chemotactic response. PMID- 6278469 TI - Spin trapping of free radicals during hepatic microsomal lipid peroxidation. AB - We have used spin-trapping techniques to identify radical species formed during the NADPH-stimulated peroxidation of rat hepatic microsomes. Using 5,5-dimethyl-1 pyrroline-1-oxide, we have confirmed the presence of substantial quantities of superoxide but found evidence for the formation of only small quantities of hydroxyl radical. Use of the spin traps N-tert-butyl-alpha-4-pyridylnitrone-1 oxide and 2-methyl-2-nitroso-1-propanol have allowed us to determine that lipid peroxyl radicals are the predominant "lipid-type" radical found in peroxidizing microsomes under aerobic conditions. These data suggest that microsomal lipid dienyl radicals react with molecular oxygen at near diffusion-controlled rates. PMID- 6278470 TI - A DNA fragment with an alpha-phosphorothioate nucleotide at one end is asymmetrically blocked from digestion by exonuclease III and can be replicated in vivo. AB - 2'-Deoxyadenosine 5'-O-(1-thiotriphosphate) (dATP[alpha S]) was introduced into the 3' ends of DNA restriction fragments with Escherichia coli DNA polymerase I to give phosphorothioate internucleotide linkages. Such "capped" 3' ends were found to be resistant to exonuclease III digestion. Moreover, the resistance to digestion is great enough that, under conditions used by us, just one strand of a double helix is digested by exonuclease III when a cap is placed at only one end; when digestion is carried to completion, this results in production of intact single strands. When digestion with exonuclease III is limited and is followed by S1 nuclease treatment, double-stranded DNA fragments asymmetrically shortened from just one side are produced. In this was thousands of nucleotides can be selectively removed from one end of a restriction fragment. In vitro introduction of phosphorothioate linkages into one end of a linearized replicative plasmid, followed by exonuclease III and S1 nuclease treatments, gives rise to truncated forms that, upon circularization by blunt-end ligation, transform E. coli and replicate in vivo. PMID- 6278471 TI - Enzymatic replication of the origin of the Escherichia coli chromosome. AB - An enzyme system that replicates plasmids bearing the origin of the Escherichia coli chromosomes (oriC) has the following physiologically relevant features. The system (i) depends completely on low levels of exogenously furnished supercoiled oriC plasmids, (ii) uses only those plasmids that contain the intact oriC region of about 245 base pairs, (iii) initiates replication within or near the oriC sequence and proceeds bidirectionally, (iv) proceeds linearly, after a 5-min lag, for 30-40 min to produce as much as a 40% increase over the input DNA, (v) depends on RNA polymerase and gyrase as indicated by total inhibition by rifampicin and nalidixate, (vi) depends on replication proteins (e.g., dnaB protein and single-stranded DNA binding protein) as judged by specific antibody inhibitions, (vii) operates independently from protein synthesis, and (viii) depends on dnaA activity, as suggested by the inactivity of enzyme fraction from each of two dnaA temperature-sensitive mutant strains, and complementation (with a 15-fold overproduction of complementing activity) by a fraction from a strain containing the dnaA gene cloned in a multicopy plasmid. Resolution and analysis of factors that control the initiation of a chromosome cycle should become accessible through its enzyme system. PMID- 6278472 TI - Salts promote activation of fat cell adenylate cyclase by GTP: special role for sodium ion. AB - The effects of GTP on adenylate cyclase [ATP pyrophosphate-lyase (cyclizing), EC 4.6.1.1] of human and rat fat cell membranes ("ghosts" and purified membranes) were examined in the absence and presence of added inorganic salts. With human ghosts GTP alone (0.1 mM) inhibited enzyme activity by 40% at 30 degrees C and had no significant effect at 37 degrees C. At both temperatures Na+ salts of Cl-, N3-, and SO2-(4) stimulated activity (up to 4-fold basal activity for 200 mM NaN3), with maximal effects at salt concentrations of 100-200 mM. Over the same concentration range these salts also allowed temperature-dependent stimulation by GTP. GTP increased the maximal activity produced by salt alone by about 2-fold at 30 degrees C and about 4-fold at 37 degrees C. Na+ (added as Cl-) was much more effective than other alkali metal cations in promoting activation by GTP. Na+ salts allowed activation of the human enzyme by the GTP analog 5'-guanylyl imidodiphosphate and also promoted stimulation of rat fat cell adenylate cyclase by both nucleotides. In time course studies of human and rat fat cell ghosts, GTP appeared to sustain an initial high rate of salt-stimulated activity, which in the absence of nucleotide subsequently fell to a lower rate, suggesting that salts might activate adenylate cyclase by promoting the stimulatory effect of endogenous membrane-bound GTP. However, with purified human fat cell membranes and a GTP-free system, salts were still stimulatory and promoted activation by added GTP. These results differ from those of previous reports in other systems in which Na+ has promoted only inhibitory effects in GTP regulation of adenylate cyclase. PMID- 6278473 TI - Method for studying kinetics of light-induced transport across membranes. AB - A technique based on phase spectrophotometry is described for studying the rates of elementary processes associated with the light-driven transport of ions and molecules across membranes. The light-induced pumping of protons by bacteriorhodopsin reconstituted into phospholipid vesicles and by chloroplast thylakoids has been studied to illustrate the potential of this technique. The exciting light is modulated by a mechanical chopper over the frequency range 5 Hz to 2 kHz. The internal pH of the membrane vesicles is modulated at the same frequency as the exciting light but differs in phase because of the finite rate of proton pumping. Measurement of this phase difference or of the frequency dispersion of the amplitude of the internal pH modulation is accomplished by use of a lock-in amplifier. The results can be interpreted in terms of relaxation times characterizing the chemical steps in proton pumping. The shortest relaxation time that can be measured is about 50 microseconds, although the time resolution could be easily extended by use of faster light chopping techniques. At pH 8.0, two relaxation processes are associated with proton pumping by bacteriorhodopsin reconstituted into phospholipid vesicles; the relaxation times are 2 and 28 msec. Two relaxation processes also were found with chloroplast thylakoids at pH 7.8, with relaxation times of 2 and 16 msec. The former can be associated with photosystem II and the latter, with photosystem I. PMID- 6278474 TI - Characterization of corticotropin receptors on adrenocortical cells. AB - The binding of corticotropin (ACTH) to receptors on isolated rat adrenocortical cells was investigated with the aid of [[125I]ITyr23, Phe2, Nle4]ACTH-(1-38) (125I-ACTH analog) which retained full biological potency and had a specific radioactivity of 1800 +/- 75 Ci/mmol. Binding was highly specific to adrenocortical cells, and the radioactive peptide was displaced by low concentrations of ACTH but not by other basic peptides. Binding was rapid, reversible, and linearly related to the number of cells. 125I-ACTH analog was not significantly degraded by incubation with the cells at 23 degrees C for 1 hr. Scatchard analysis of the binding was compatible with a single class of binding sites with Kd = 1.41 +/- 0.21 nM, and the number of sites was estimated to be 3840 +/- 1045 per cell. The binding curve was superimposable on the concentration response curve for cyclic AMP. Small, but significant amounts of 125I-ACTH analog were bound at concentrations sufficient for maximal stimulation of steroidogenesis. For a series of ACTH analogs, the concentrations of the peptides required for half-maximal stimulation of cyclic AMP production were in excellent agreement with the concentration required for half-maximal inhibition of binding. These results suggest that the adrenocortical cells contain only one class of ACTH receptors and that stimulation of a small fraction of these receptors (less than 3%) is sufficient for maximal steroidogenesis. PMID- 6278475 TI - Prolactin-like activity of anti-prolactin receptor antibodies on casein and DNA synthesis in the mammary gland. AB - Prolactin receptors were partially purified from rabbit mammary gland membranes by using an affinity chromatography technique. Antibodies against this prolactin receptor preparation were obtained in guinea pig and sheep. Both antisera were able to inhibit the binding of 125I-labeled ovine prolactin to rabbit mammary gland membranes. When added to culture media of rabbit mammary explants, the anti prolactin receptor antiserum inhibited the capacity of prolactin to initiate casein synthesis and casein mRNA accumulation as a function of the antiserum concentration. However, in the absence of prolactin, both antisera (guinea pig and sheep) at moderate concentrations were capable of mimicking prolactin action on casein gene expression and on DNA synthesis. At higher concentrations, the anti-prolactin receptor antibodies inhibited their own actions. Several characteristics of the prolactin effect were also observed with the anti prolactin receptor antibody: the stimulatory effect of the antibody was amplified by glucocorticoids; colchicine, which was capable of blocking prolactin action, also prevented the induction by the antibody. Lysosomotropic agents, which do not interfere with prolactin action, did not alter the response observed with the antibody. These results indicate that an anti-prolactin receptor antibody can mimic two major actions of prolactin obtained in mammary explant culture and suggests that the prolactin molecule is not required beyond the initial binding to its receptor. PMID- 6278476 TI - Transmembranous incorporation of photoelectrically active bacteriorhodopsin in planar lipid bilayers. AB - Various methods to incorporate bacteriorhodopsin in black lipid membranes are reported. Both purple membrane patches and monomeric bacteriorhodopsin were used as starting material. The incorporation of bacteriorhodopsin into planar lipid bilayers was achieved by the following methods. (i) Purple membrane patches were transferred from water to solutions of lipids in n-alkanes. Black membranes were formed from such organic suspensions. (ii) Lipid layers containing solvent and purple membranes were spread on an air/water interface. These layers were used to form planar bilayers. (iii) Vesicles containing purple membranes or monomeric bacteriorhodopsin were spread on an air/water interface and, from the resulting layer, bilayers were formed. On illumination, steady-state photocurrents were observed in all three cases, indicating that these methods lead to functional transmembranous integration of the protein in the planar black lipid membrane. The influence of an applied electric field on the pumping process was studied on membranes formed by using method i. At approximately 200 mV, the photocurrent tends to zero. Furthermore, it was possible to make planar lipid bilayers photoelectrically active by adding vesicles containing monomeric bacteriorhodopsin to the bathing solution. Because, in this case, only transient photocurrents were observed, it can be concluded that the vesicles are attached to but not fused with the black lipid membrane. PMID- 6278477 TI - Primary step in the bacteriorhodopsin photocycle: photochemistry or excitation transfer? AB - The absorption polarization of the first intermediate (K610) formed at room temperature in the proton-pumping photochemical cycle of bacteriorhodopsin (bR) shows a strong correlation with the polarization direction of the photolyzed parent molecule. The results suggest that, unlike other photosynthetic systems, excitation transfer does not take place prior to the primary photochemical change in bR. These observations together with the previously observed circular dichroism and the polarization temperature dependence are discussed in terms of the exciton structure and the nature of the absorption bandwidths (i.e., homogeneous vs. inhomogeneous) of the bR monomers within the trimer structure. PMID- 6278478 TI - Monoclonal antibodies against receptor for epidermal growth factor induce early and delayed effects of epidermal growth factor. AB - Mice were immunized with human epidermoid carcinoma cells (A-431 cell line) that possess an unusually high number of membrane receptors for epidermal growth factor (EGF). Spleen cells from these mice were fused with NSI cells, a nonsecreting murine myeloma. The immunoglobulins secreted by the obtained hybridomas were screened for specific binding to A-431 cells and selected according to their ability to inhibit the binding of radiolabeled EGF to the membrane of A-431 cells. Several antibodies secreted by cloned hybrid lines were found to inhibit the binding of radiolabeled EGF to membrane receptors of living A-431 cells, human foreskin fibroblasts, and mouse 3T3 fibroblasts and also to membrane preparations from A-431 cells. These monoclonal antibodies induced the early and delayed biological effects mediated by EGF. Like EGF, the antibodies induced morphological changes in A-431 cells and enhanced the phosphorylation of endogenous membrane proteins in membranes from these cells. They also stimulated DNA synthesis in human foreskin fibroblasts. These observations support the notion that the biological information of the EGF-receptor complex resides in the membrane receptor. Furthermore, the antibodies offer a powerful tool to study the structure, processing, and mode of action of EGF receptors. PMID- 6278479 TI - Efficient infection of monkey cells with DNA of simian virus 40. AB - With standard protocols for DNA infection, only a small fraction (about 4%) of monkey cells exposed to purified DNA of simian virus 40 (SV40) exhibits signs of infection. We have devised a protocol by which we can extend the time of exposure of BSC-1 cells to DNA in the presence of low concentrations of DEAE-dextran. The efficiency of infection is proportional to the time of exposure. With an 8-hr exposure, we are reproducibly able to infect 25% of the cells, and we have been able to achieve levels of infection as high as 50% with a 16-hr exposure. The percentage of cells infected was measured either by scoring for nuclei positive for SV40 tumor antigen or by an infectious centers assay. We also report the use of ethidium bromide as a nonspecific nuclear counterstain in the immunofluorescence assay for SV40 tumor antigen. PMID- 6278480 TI - Immunohistochemical localization of renin in luteinizing hormone-producing cells of rat pituitary. AB - The location of renin (EC 3.4.99.19) in rat pituitary was determined by the peroxidase-antiperoxidase immunohistochemical technique. By using antisera prepared with purified rat renal renin, an immunoreactive substance was localized within ovoid cells scattered throughout the anterior pituitary. These cells were shown to be luteinizing hormone-producing cells by staining with anti-luteinizing hormone antisera in adjacent sections. By using the double staining method, the renin-containing cells were differentiated from cells containing corticotropin, thyrotropin, growth hormone (somatotropin), and prolactin (mammotropin). These results suggest a possible local role for renin in the anterior pituitary. PMID- 6278481 TI - Rapid glucocorticoid inhibition of vasoactive intestinal peptide-induced cyclic AMP accumulation and prolactin release in rat pituitary cells in culture. AB - Vasoactive intestinal peptide (VIP) stimulates both adenosine 3',5'-cyclic monophosphate (cAMP) accumulation and prolactin release in normal rat pituitary cells in culture. cAMP accumulation is significant (P less than 0.01) at VIP concentrations as low as 1 nM and reaches a maximum with 0.1 microM. Addition of dexamethasone as early as 15 min before VIP inhibits VIP stimulation of both cAMP production and PRL secretion. The rapid inhibition is dose-dependent: it appears at doses as low as 0.01 pM and is complete at 1 pM dexamethasone. Increasing concentrations of dexamethasone induce a noncompetitive type of inhibition, as shown by the decrease in Vmax with no change in the apparent Km for VIP. Cycloheximide (1 mM) counteracts the inhibitory effect of dexamethasone on VIP induced cAMP production, which suggests the involvement of a rapid protein synthesis mechanism. Ru-26988, a specific glucocorticoid devoid of any mineralocorticoid activity and which does not bind to intracellular transcortin like component, also produces an inhibition of VIP-induced cAMP accumulation. Corticosterone also inhibits VIP-induced cAMP production but at concentrations higher than those of dexamethasone. In contrast, aldosterone, progesterone, estradiol, and testosterone have no effect. These results demonstrate that, in normal rat pituitary cells in culture, glucocorticoids at physiological concentrations rapidly inhibit the cAMP production and prolactin release induced by VIP by acting through specific glucocorticoid receptors. PMID- 6278482 TI - Rat liver gap junction protein: properties and partial sequence. AB - Gap junctions, strongly implicated as channels for direct cell-to-cell communication, have been isolated from rat liver in high yield and purity. These gap junction fractions contain few morphologically recognizable contaminants, but NaDodSO4/polyacrylamide gel electrophoresis reveals a number of polypeptides. With the exception of a nonjunctional component of Mr 38,000 and some poorly soluble material, including collagen, all the polypeptides have very similar or identical two-dimensional peptide maps and arise from proteolytic cleavage of the COOH-terminus or aggregation of a Mr 28,000 protein. We report the sequence of the NH2-terminal 52 amino acids of this protein. The polypeptide (Mr approximately equal to 10,000) characteristic of trypsin-treated gap junction preparations is shown to be two distinct polypeptides, both derived from the Mr 28,000 protein. PMID- 6278483 TI - Activation of lymphocytes by brominated nucleoside and cyclic nucleotide analogues: implications for the "second messenger" function of cyclic GMP. AB - The purine nucleoside guanosine, when derivatized at the C-8 position to give 8 bromoguanosine (8-BrGuo), acquires the capacity to stimulate high-level lymphocyte proliferation in the presence or absence of serum. Direct comparisons were undertaken to determine whether this activity is exerted only by virtue of the structural resemblance of 8-BrGuo to 8-bromo cyclic GMP (8-BrcGMP) (a known intracellular lymphocyte mitogen). They showed that, of the brominated guanosine derivatives studied, 8-BrGuo is the primary activator because (i) it is a far more potent lymphocyte activator than 8-BrcGMP, the order of mitogenic potency being 8-BrGuo greater than 8-bromo GMP (8-BrGMP) greater than 8-BrcGMP; (ii) it acts much more rapidly than 8-BrcGMP; (iii) it is not metabolized to 8-BrcGMP or cGMP; and (iv) it does not elevate intracellular cGMP content. cGMP is not likely to be the second messenger serving to activate B cells because (i) it does not induce significant proliferation unless brominated at the C-8 position; (ii) the brominated form is much less efficient than 8-BrGuo or 8-BrGMP; (iii) 8-BrGuo and many other mitogens do not increase intracellular cGMP; (iv) many agents that increase cGMP fail to initiate lymphocyte activation; (v) certain agents that increase cGMP (i.e., 15-hydroperoxyarachidonic acid, azide) inhibit lymphocyte activation; and (vi) addition of unbrominated cGMP to cultures stimulated with 8 BrGuo actually diminished stimulation. These data (i) indicate that, by interaction with cellular components, 8-BrGuo triggers high level lymphocyte activation and (ii) cast significant doubt on the role of cGMP as an intracellular second messenger in lymphocyte proliferation. PMID- 6278484 TI - Mutations in the lacY gene of Escherichia coli define functional organization of lactose permease. AB - Mutations in the lacY gene of Escherichia coli have been used to analyze the functional organization of lactose permease. Deletions suggest that the NH2 terminus of lactose permease is not essential and can be replaced by residues of the cytoplasmic enzyme beta-galactosidase. Negative dominant mutations in the lacY gene can be explained by the assumption that membrane-associated lactose permease is active as a dimer or oligomer. The map positions of these mutations and other point mutations that lower or alter the sugar specificity define regions of lactose permease involved in sugar or proton binding and transport. PMID- 6278485 TI - Down syndrome fibroblasts are hyperresponsive to beta-adrenergic stimulation. AB - The hormonal response to human skin fibroblasts after exposure to beta-adrenergic agonists, prostaglandin E1 (PGE1), and cholera toxin was monitored by intracellular cyclic AMP accumulation. Down syndrome (DS; trisomy 21) cells had an approximately 10-fold greater response to beta-adrenergic agonists than did either normal diploid skin fibroblasts or other aneusomic fibroblast strains (trisomy 13, 18, and 22). The altered response in DS fibroblasts was specific for beta-adrenergic agonists, because treatment of DS or control cells with PGE1 or cholera toxin resulted in the same degree of cyclic AMP accumulation. Experiments with 3-isobutyl-1-methylxanthine, a cyclic nucleotide phosphodiesterase inhibitor, indicated that the increased response of DS fibroblasts was not primarily a function of altered cyclic AMP degradation. Monosomy 21 cells responded less than normal diploid fibroblasts to stimulation by the beta adrenergic agonist isoproterenol. These findings suggest that genetic information on chromosome 21 participates in regulating the beta-adrenergic response of human fibroblasts. PMID- 6278486 TI - Anti-poly(A) polymerase antibodies in sera of tumor-bearing rats and human cancer patients. AB - Poly(A) polymerase (polynucleotide adenylyltransferase; ATP:polynucleotide adenylyltransferase, EC 2.7.7.19) was covalently linked to diazobenzyloxymethyl filters and used to screen the sera from a number of tumor-bearing rats and human cancer patients for antibodies to poly(A) polymerase. Sera from rats that had been inoculated with any of several Morris hepatomas or a mammary adenocarcinoma contained immunoglobulins capable of complexing with poly(A) polymerase. No antibodies to the enzyme could be detected in sera from control animals or from those bearing tumors for short periods of time. Antibodies to poly(A) polymerase were also observed in sera from human patients with leukemia, polycythemia vera, and Wilms tumor. The antibodies were not evident in sera from normal volunteers or from patients with nonneoplastic diseases. These included lupus erythematosus, a disorder in which antibodies are produced against an array of nuclear proteins. Immunoglobulins from the serum of one of the human patients were capable of inhibiting poly(A) polymerase activity in vitro, whereas those prepared from the serum of a normal volunteer did not affect enzyme activity. As determined by the diazobenzyloxymethyl-filter technique, the relative concentration of antibodies in the sera of an individual with leukemia (in remission) increased severalfold during a relapse. These data suggest that the presence of antibodies to poly(A) polymerase may be characteristic of sera from cancer patients and that the relative concentration of these antibodies may be indicative of the disease state. PMID- 6278487 TI - Human chorionic gonadotropin radioantibodies in the radioimmunodetection of cancer and for disclosure of occult metastases. AB - Radioimmunodetection (RaID) of tumors containing human chorionic gonadotropin (hCG; choriogonadotropin) was evaluated in 25 patients by injecting 131I-labeled goat antibody IgG against hCG and performing total-body photoscans with a gamma scintillation camera 24 and 48 hr later. All 10 testicular cancer patients with proven tumor sites had positive RaID results, whereas three cases without known tumor were negative. Four patients with hydatidiform mole and one with degenerative products of conception showed positive RaID results consistent with elevated serum hCG titers. Two putatively false-positive results were obtained in patients with lung or ovarian cancer, whereas a false-negative metastasis to the liver of a patient with lung cancer and an elevated serum hCG titer was observed. Of 14 tumor sites found by RaID in 10 testicular cancer patients, 4 were revealed by RaID prior to any other detection method used and provided a lead time to definitive diagnosis by other measures of a few days to greater than 1 yr. Although a number of patients had high serum hCG levels, even exceeding 3 microgram/ml, the xenogeneic antibody was capable of localizing in tumor. No adverse effects were noted in any of the patients studied. Thus, hCG RaID appears to be a safe and effective method of detecting and locating hCG-producing tumors and has been found to disclose occult testicular cancers. PMID- 6278488 TI - The U3 portion of feline leukemia virus DNA identifies horizontally acquired proviruses in leukemic cats. AB - The presence and location of DNA sequences related to the U3 and U5 portions of the infectious exogenous feline leukemia virus (FeLV) long terminal repeat (LTR) in various cat DNAs have been determined by hybridization experiments. In uninfected cat DNAs, the U5 LTR segment from the Gardner-Arnstein strain B virus is present at approximately 150 copies per cell. This level is approximately 10 fold greater than that of endogenous internal FeLV sequences. The U5 sequences differ in copy number and, to some extent, in location from one animal to another. For any one animal, the sequence organization of the U5 segments is the same among different tissues, showing that the pattern is inherited through the germ line. Most importantly, the viral U3 LTR probe hybridizes only very weakly with uninfected cat DNAs. Both the U3 and the U5 regions of the LTR from the Gardner-Arnstein strain of virus cross-hybridize with DNA derived from four other infectious FeLVs representing A, B, and C subtypes. Thus, the C3 region may be used as a probe for studying the number and location of exogenously acquired FeLV proviruses in infected cat tissues. In some cases exogenously acquired proviruses are present in unique sites in the genome of virus-positive cat lymphosarcomas, indicating a monoclonal origin for the tumor. In other tumors, the proviral sequences are randomly distributed over many sites. Lymphosarcomas of virus negative cats have no exogenous U3 sequences despite epidemiological evidence of an association of virus-negative leukemia with exposure to FeLV. PMID- 6278489 TI - The behavioral effects of phencyclidines may be due to their blockade of potassium channels. AB - The action of phencyclidine [1-(1-phenylcyclohexyl)piperidine; PCP] and its behaviorally active analog (m-amino-PCP) and of two behaviorally inactive analogs [m-nitro-PCP and 1-piperidinocyclohexanecarbonitrile (PCC)] were examined in this study. In a test of spatial alternation performance in rats, PCP and m-amino-PCP were much more potent behavior modifiers than were PCC and m-nitro-PCP. We studied the effects of the drugs on the ionic channels of the electrically excitable membrane and of the nicotinic acetylcholine (AcCho) receptors at the neuromuscular junction of frog skeletal muscle. All four compounds blocked the indirectly elicited muscle twitch and depressed the amplitude and rate of rise of directly elicited muscle action potentials. They also caused a voltage- and concentration-dependent decrease in the peak amplitude of the endplate current but did not react with the nicotinic AcCho receptor. These observations indicate that the four compounds have comparable blocking effects on the ionic channels associated with the nicotinic AcCho receptor. In contrast, the behaviorally active agents could be distinguished from behaviorally inactive ones by their effects on K+ conductance. PCP and m-amino-PCP blocked delayed rectification in frog sartorius muscles, prolonged the muscle action potential more than 2-fold, and markedly potentiated the directly elicited muscle twitch. The behaviorally active compound also blocked depolarization-induced 86Rb+ efflux from rat brain synaptosomes (presumably a measure of K+ conductance) and increased quantal content at the frog neuromuscular junction. In these actions, m-nitro-PCP was much less effective, and PCC was relatively ineffective. Because PCP and m-amino PCP are much more potent behavior modifiers than PCC and m-nitro-PCP, we suggest that the behavioral effects of PCP and m-amino-PCP, may be due to a block of K+ conductance and enhancement of transmitter release at central neurons. PMID- 6278490 TI - Changes in neuromuscular junction endplate current time constants produced by sulfhydryl reagents. AB - The acetylcholine receptor is a protein that contains certain critical disulfide bonds. Experiments were designed to determine the role such bonds might play in the physiological activity of the receptor. Modification of the receptor with sodium bisulfite and diamide produced an increase in the time constants of the miniature endplate current without changes in the single-channel properties of the receptor. Controls were done to determine that this change in the miniature endplate current was not due to an effect on acetylcholinesterase at the endplate. These data are interpreted to mean that the reagents increase the time acetylcholine is bound to the receptor before the channel opens and is most probably due to a change in receptor affinity brought about by chemical modification of the receptor protein. PMID- 6278491 TI - Isolated horizontal cells from carp retina demonstrate dopamine-dependent accumulation of cyclic AMP. AB - Horizontal cells of the carp retina were separated from other retinal cell types by using enzymatic dissociation and velocity sedimentation at unit gravity. Fractions containing horizontal cells were tested for their ability to accumulate cyclic AMP in the presence of various putative neurotransmitters. Micromolar concentrations of dopamine, when added in the presence of 3-isobutyl-1 methylxanthine, stimulated cyclic AMP accumulation in these isolated cells. The dopamine-dependent accumulation of cyclic AMP in intact isolated horizontal cells was blocked by nanomolar concentrations of dopamine antagonists such as haloperidol, (+)-butaclamol, and fluphenazine. The results indicate that there is a postsynaptic dopamine receptor on carp horizontal cells that is associated with adenylate cyclase [ATP pyrophosphate-lyase (cyclizing), EC 4.6.1.1]. PMID- 6278492 TI - Simple rapid method for the synthesis of radioactively labeled cDNA hybridization probes utilizing bacteriophage M13mp7. AB - Double-stranded cDNA sequences for rat alpha 1-acid glycoprotein and rat glutathione S-transferase mRNAs were inserted into the Pst I site of bacteriophage M13mp7 and used to develop a new method for preparing specific cDNA hybridization probes directly from cloned template DNA. A palindrome sequence surrounding the Pst I site in the vector DNA permitted single-stranded DNA isolated from the recombinant phage to fold back, thus forming a stable hybrid bounded on the ends by a large loop of M13mp7 single-stranded DNA and a small loop of inserted foreign DNA. A primer corresponding to an internal sequence of the foreign DNA was hybridized, then Escherichia coli DNA polymerase I was used to synthesize a 32P-labeled complementary DNA copy of the cloned inserted DNA. The single-stranded cDNA reaction product was easily isolated by subsequent sedimentation through alkaline sucrose gradients. Gel electrophoresis of the labeled cDNA product, after denaturation with glyoxal, indicated a single discrete band with an electrophoretic mobility corresponding to the length of the inserted DNA sequence. About 95% of the cDNA product formed S1 nuclease-resistant hybrids in hybridization reactions with excess RNA in solution. DNA sequences complementary to the M13mp7 vector DNA were not detected in the cDNA product. Thus, these M13mp7-derived probes are the functional equivalent of cDNA copies to mRNAs and can be employed for quantitative measurements of mRNA concentration. This simple, rapid method probably can be used for most cloned DNA sequences to yield single-stranded radioactively labeled DNA, without contaminating vector DNA sequences, for virtually any hybridization requirement. PMID- 6278493 TI - Cytochrome c gene-related sequences in mammalian genomes. AB - We use a rat cytochrome c gene that we previously isolated and determined the sequence of to estimate the number of related sequences present in the rat genome. Approximately 25 different EcoRI restriction endonuclease fragments from total rat DNA hybridize to the gene of known structure. Four of these correspond to homologous sequences present in four different lambda Charon 4A-rat cytochrome c recombinants previously isolated. Intact or nearly intact genes appear to reside on almost all of the genomic fragments, because they hybridize strongly to gene subfragments representing both 5' and 3' portions of the coding sequence as well as to 3' noncoding DNA that is found specifically associated with the coding region. A subgroup of about six of the fragments also shares homology within the 73 nucleotides immediately preceding the AUG codon. An intron-specific probe reveals only the EcoRI fragment from which it was derived and one other genomic fragment. On the basis of the temperature of complete dissociation of the coding region probe in 0.75 M NaCl/0.075 M Na3 citrate/50% (vol/vol) formamide, the 25 fragments are separable into three stringency classes of 40-50 degrees C, 50-55 degrees C, and 55-60 degrees C. The latter, high-stringency group of about seven fragments includes those cloned in the recombinant phage isolates, whose regions homologous to cytochrome c are shown to differ from the purified gene of known sequence by an amount equivalent to about 2% mismatched bases. Families of cytochrome c gene-related sequences are also found in the genomes of several other mammals, including humans. PMID- 6278494 TI - Variable-sized free episomes of Shope papilloma virus DNA are present in all non virus-producing neoplasms and integrated episomes are detected in some. AB - The state of rabbit (Shope) papilloma virus DNA in virus-induced nonproducing tumors on domestic rabbits was investigated. Virus-specific sequences were resolved into many distinct bands by one-dimensional agarose gel electrophoresis. Two-dimensional gel electrophoresis, CsCl/propidium iodide density equilibrium centrifugation, partial digestion with a restriction endonuclease, and S1 nuclease digestion permitted us to identify the bands as free viral episomes representing circular molecules of increasing size. In some tumors (both papillomas and carcinomas), up to 25% of the virus-specific DNA was linear and comigrated with cellular DNA. Integration of at least some of these sequences was suggested by the detection of viral-cellular junction bands in one tumor after digestion of DNA with EcoRI and Sal I, enzymes that cut Shope DNA once. Finally, the physical states of viral DNA in papillomas and carcinomas were found to be similar, although free episomes were generally larger in carcinomas. PMID- 6278496 TI - Reconstitution in planar lipid bilayers of a Ca2+-dependent K+ channel from transverse tubule membranes isolated from rabbit skeletal muscle. AB - Addition of membrane vesicles prepared from transverse tubule (T-tubule) membranes of rabbit skeletal muscle to the aqueous phase of a planar lipid bilayer induces a stepwise increase in conductance. This conductance is both voltage and Ca2+ dependent. At 1 mM Ca2+, the steady-state conductance is maximal at voltages higher than +20 mV and decreases for more negative voltages. (Voltages refer to the side to which the vesicles are added, cis) Decreasing the Ca2+ concentration reversibly shifts the conductance-voltage curve toward the right along the voltage axis. Furthermore, Ca2+ can activate the conductance only if added to the cis compartment. Neither Mg2+, Ba2+, nor Cd2+ can activate the conductance induced by T-tubule vesicles. Addition of 5 mM tetraethylammonium ion to the trans, but not the cis, side abolishes the T-tubule-induced conductance. The Ca2+-dependent conductance appears as a consequence of ionic channel formation. Single-channel activity appears in bursts followed by periods of time in which the channel remains "silent". The conductance of the open channel averages 226 pS in 0.1 M KC1 and is voltage and Ca2+ independent. However, the fraction of time that the channel remains in the open state is voltage and Ca2+ dependent in a manner that parallels the voltage and Ca2+ dependence of the multichannel membrane. The channel is 6.6 times more permeable to K+ than to Na+ and is impermeable to C1-. PMID- 6278495 TI - Expression of transforming region of Moloney murine sarcoma virus in Escherichia coli as a fusion protein with small tumor antigen of polyoma virus. AB - Bacterial expression of the transforming region of Moloney murine sarcoma virus, designated mos, was obtained as a fusion protein with a portion of the small tumor antigen of polyoma virus. This was accomplished by fusing the entire mos open reading frame, encoding a 41,000-dalton protein, with a plasmid that expresses a beta-galactosidase-polyoma fusion protein under lac operon control. The resulting plasmid directed synthesis of the predicted polyoma antigen-sarcoma virus fusion protein of 59,000 daltons. This protein was immunoprecipitated by an anti-polyoma tumor antigen antiserum that recognized polyoma determinants at the NH2 terminus of the hybrid protein. This protein was also immunoprecipitated by an antiserum directed against a synthetic peptide containing the 12 COOH-terminal amino acids encoded by the mos open reading frame. This work confirms the existence of a long open reading frame in the mos gene and resolves a discrepancy between different nucleotide sequences for its COOH-terminal coding region. PMID- 6278497 TI - Increase in apparent compressibility of cytochrome c upon oxidation. AB - The apparent molal adiabatic compressibilities of ferri- and ferrocytochrome c have been determined from measurements of density and sound velocity. The values found were +2.99 X 10(-8) and -2.40 X 10(-8) cm5 mol-1 dyne-1 for the ferri and ferro forms, respectively. Experiments were performed on identical solutions containing either the oxidized or reduced form of protein. Solutions of ferricytochrome c were found to have significantly greater adiabatic compressibility than equivalent solutions of ferrocytochrome c at 25 degrees C and pH 7.15. The remarkable similarity of the three-dimensional structures of the ferri and ferro proteins [Takano, T. & Dickerson, R.E. (1980) Proc. Natl. Acad. Sci. USA 77, 6371-6375] strongly suggests that this difference in compressibility is due to an increase in volume fluctuations within ferricytochrome c relative to the ferro form rather than a change in equilibrium structure or hydration. Such a difference in the dynamic properties of the structures is consistent with both the crystallographic thermal B factors and the observed increase in amide hydrogen exchange kinetics when ferrocytochrome c is oxidized. The relative magnitude of the root mean square volume fluctuations is approximated from an ideal solution treatment of the compressibility data and yields a ratio of delta Vrms (ferri cyt c)/ delta Vrms (ferro cyt c) = 1.3. PMID- 6278498 TI - Cyclic AMP-mediated control of meiosis: effects of progesterone, cholera toxin, and membrane-active drugs in Xenopus laevis oocytes. AB - Progesterone depressed rapidly (50% at 1 min) and persistently cyclic AMP (cAMP) concentration that had been elevated by cholera toxin in Xenopus laevis oocytes. cAMP remained below 1 pmol per oocyte (mean basal level) for approximately 1 hr and thereafter rose to approximately 120% of control values, while germinal vesicle (nucleus) breakdown did not occur. In the absence of cholera toxin, progesterone treatment for 6 hr maintained cAMP concentration below the basal level (but not lower than 80%), and germinal vesicle breakdown occurred. Experiments in the presence of phosphodiesterase inhibitors suggested that progesterone modulates adenylate cyclase activity. The maturation promoting factor, which is formed after 3-5 hr of progesterone treatment and provokes germinal vesicle breakdown after its injection into untreated oocytes, also decreased cAMP concentration, an observation that may explain its "autoamplification." Nonsteroidal inducers of meiosis reinitiation (e.g., propranolol, methoxyverapamil, mersalyl) diminished the cholera toxin-mediated accumulation of cAMP, in contrast to compounds devoid of meiotic-inducing capacity and antagonist to progesterone action, such as gammexane (an inositol analogue) and 5'-deoxy-S-(2-methylpropyl)-5'-thioadenosine (a methylase inhibitor), that increased the nucleotide level. The fine control, suggested by the effects of small changes in cAMP levels, gives evidence of great sensitivity to a critical determinant governing meiotic cell division. PMID- 6278499 TI - Plasmid gene organization: naphthalene/salicylate oxidation. AB - Genes for naphthalene metabolism are localized on nah7, an 83-kilobase (kb) plasmid, in two gene clusters under salicylate control. Polar mutations formed by insertion of the transposon Tn5 permit detection of the transcription direction and the gene organization within two approximately 10-kb DNA segments separated by a approximately 7-kb regulatory gene region. The gene cluster specifying conversion of naphthalene to salicylate lies near the left initiation of a 25-kb DNA fragment A released by EcoRI; that for the salicylate pathway via catechol meta-fission lies near the right terminus with extension into the adjoining 5.9 kb fragment C. The genetic organization and regulation resemble the tol plasmid encoded "upper" and "lower" pathways of toluene/xylene oxidation in Pseudomonas putida mt2. PMID- 6278500 TI - Lymphocyte-conditioned medium protects human monocyte-macrophages from cholesteryl ester accumulation. AB - Exposure of human monocyte-macrophages to as little as 50 microliters of cultured medium from lymphocytes stimulated by concanavalin A (Con A) resulted in a dramatic decrease in the activities of the low density lipoprotein (LDL) receptor pathway, the LDL-dextran sulfate pathway, and the scavenger receptor pathway. This effect was not seen when the monocyte-macrophages were exposed to culture medium from lymphocytes cultured without Con A or with Con A together with alpha methyl mannoside or control medium without lymphocytes. The activity of 3-hydroxy 3-methyglutaryl-coenzyme A reductase also decreased in monocyte-macrophages exposed to culture medium from stimulated lymphocytes. Acyl-CoA:cholesterol O acyltransferase activity, protein synthesis, protein content, phagocytosis of heat-killed yeast, and non-receptor-mediated endocytosis were not inhibited. Monocyte-macrophages exposed to malondialdehyde altered-LDL in the presence of stimulated lymphocyte culture medium accumulated substantially less cholesteryl esters than did cells in control medium. We propose that substances produced by stimulated lymphocytes may be useful in protecting macrophages from cholesteryl ester accumulation. PMID- 6278501 TI - Ca-mediated activation of a K current at fertilization of golden hamster eggs. AB - Golden hamster eggs respond to fertilization with recurring hyperpolarizations [Miyazaki, S. & Igusa, Y. (1981) Nature (London) 290, 703-705]. We analyzed the ionic mechanism of the fertilization potential and examined whether the fertilization potential plays a role in polyspermy block. Each hyperpolarizing response (HR) during fertilization is found to be caused by an increase in the K conductance activated by an increase in the intracellular Ca2+ concentration. This conclusion is based on the following: (i) The reversal potential of the HR shifted with the Nernstian slope for K ions when the external K concentration was changed, whereas it was unaltered by the removal of Cl ions. (ii) The HR was blocked by the intracellular injection of EGTA. (iii) Injection of Ca2+ into an egg induced a hyperpolarization of the membrane similar to the HR. The Ca activated K conductance shows an apparent outward rectification, which could be explained by an asymmetric distribution of K ions across the membrane. The HR associated with sperm entry into the egg occurred at any membrane potential between -160 and +50 mV. Therefore, a potential-dependent block of sperm entry does not occur in the hamster egg. PMID- 6278502 TI - Carp horizontal cells in culture respond selectively to L-glutamate and its agonists. AB - Horizontal cells were enzymatically isolated from the carp retina and maintained in culture for 2-7 days. Cultured horizontal cells typically had resting membrane potentials of -50 to -70 mV and input resistances of 100-150 m omega. The cells were treated with a number of neurotransmitter agents and their analogues. Significant responses were evoked only by 3,4-dihydroxyphenylalanine (dopamine), L-glutamate, and certain glutamate analogues. The responses to dopamine were inconsistent; most often, the membrane hyperpolarized and input resistances increased. However, highly characteristic responses to L-glutamate and its analogues, quisqualate and kainate, were observed in virtually all of the cells tested. The responses consisted of an initial graded depolarization accompanied by a resistance increase, followed in most cases by a prolonged (1- to 2-min) regenerative depolarization. The regenerative component of the response appears to be Ca2+ dependent, while the underlying graded potential may be due to a decrease in K+ conductance of the membrane. PMID- 6278503 TI - Interactions of dicyclohexylcarbodiimide with myelin proteolipid. AB - Dicyclohexylcarbodiimide (DCCD) is known to bind preferentially to a proteolipid subunit of proton-translocating systems and thereby to inhibit proton transport. In the present study we show that, in an aqueous medium, DCCD binds to the bovine white matter proteolipid apoprotein, the major protein of central nervous system myelin. The binding is dependent on time, temperature, and concentration and is not inhibited by the hydrophilic carbodiimide 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide. By contrast, when the incubation is carried out in chloroform/methanol no labeling by DCCD is demonstrable. In isolated rat myelin, DCCD binds specifically to the proteolipid and not to the myelin basic proteins. Liposomes reconstituted with the myelin proteolipid apoprotein transport protons, as assayed by quenching of the fluorescence of 9-aminoacridine. Preincubation of proteolipid-containing liposomes with DCCD results in an inhibition of transport. These studies have important implications for a possible ionophoric function of the myelin proteolipid and for the occurrence of transport processes within myelin. PMID- 6278504 TI - Photobleaching through glass micropipettes: sodium channels without lateral mobility in the sarcolemma of frog skeletal muscle. AB - Sodium currents were recorded from frog skeletal muscle by using fire-polished micropipettes to electrically isolate and voltage clamp a small patch of sarcolemma. Sodium current amplitude served as an assay for the number of functional sodium channels in the patch. With the pipette as a light guide, these channels were irradiated with ultraviolet (UV) light directed through a quartz fiber into the back end of the pipette. The UV light emerging from the pipette tip caused localized destruction of the sodium channels in the patch, reducing sodium current 3- to 5-fold during a 30-90 s irradiation. If sodium channels could diffuse laterally in the membrane, current from the patch should recover with time as fresh channels enter from neighboring areas. No such recovery was observed during observation for 1 hr after irradiation. Our results set an upper limit of 10(-12) cm2/s for the diffusion coefficient--1/1000th that of rhodopsin, a membrane protein in the cell membrane of retinal rods. It is suggested that sodium channels are anchored in the sarcolemma. PMID- 6278505 TI - Effect of vasoactive agents on fatty acid metabolism: inhibition of lipolysis by tipropidil in rat epididymal adipose tissue. PMID- 6278506 TI - Movements of benzo[a]pyrene across the hemochorial placenta of the guinea pig. PMID- 6278507 TI - Effects of a synthetic Met5-enkephalin analog on plasma luteinizing hormone and prolactin levels in conscious orchiectomized rats. PMID- 6278508 TI - beta-Adrenergic suppression of progesterone-induced luteinizing hormone surge in ovariectomized, estrogen-primed rats. PMID- 6278509 TI - Effects of solid tumors on the resistance of mice to viral and bacterial infections. PMID- 6278510 TI - Simian varicella virus (delta herpesvirus) infection of patas monkeys leading to pneumonia and encephalitis. PMID- 6278511 TI - The effect of polybrominated biphenyl (PBB) in testes, adrenal, and pituitary function in the rat. PMID- 6278512 TI - Optimization of a radioreceptor assay for human growth hormone using male rat liver membranes. PMID- 6278513 TI - Bioavailability of trace elements in human nutrition. PMID- 6278514 TI - Terminology of dietary fiber. PMID- 6278515 TI - Biochemical determinants of iron absorption. PMID- 6278516 TI - Effects of cyclic adenosine and cyclic guanosine nucleotides and theophylline on gastrin synthesis and secretion in rat antral organ culture. AB - The effect of cyclic adenosine and cyclic guanosine nucleotides and theophylline upon gastrin synthesis and secretion were examined in rat antral mucosa maintained in organ culture. In concentration-response experiments, cyclic AMP, dibutyryl cyclic AMP and theophylline stimulated gastrin release and incorporation of [3H]-tryptophan into gastrin: a high degree of positive correlation was demonstrated between gastrin synthesis and gastrin release in response to control and test culture conditions. Cyclic GMP and dibutyryl cyclic GMP also stimulated gastrin secretion to a degree similar to that seen with cyclic AMP. These studies provide support for the proposal that the cyclic nucleotide system is involved in the stimulation of gastrin synthesis and release by the antral gastrin cell. PMID- 6278517 TI - The effects of photoactivated protoporphyrin on reticulocyte membranes, intracellular activities and hemoglobin recipitation. PMID- 6278518 TI - [Comparative biochemical studies of certain urinary metabolites in patients with the nephrotic syndrome, eye diseases, alkaptonuria, obstructive myocardiopathy, and cancer and in newborn infants]. PMID- 6278519 TI - [Urine biochemistry in children with supposed metabolic diseases]. PMID- 6278520 TI - Cholecystokinin reduces exploratory behavior in mice. PMID- 6278521 TI - Melatonin and the adrenal cortex: relationship to territorial aggression in mice. PMID- 6278522 TI - Compatibility and molecular properties of plasmid Rms 149 in Pseudomonas aeruginosa and Escherichia coli. PMID- 6278523 TI - Transposition of mercury resistance from a transferable R plasmic of Escherichia coli. PMID- 6278524 TI - IS2-IS2 and IS3-IS3 relative recombination frequencies in F integration. PMID- 6278525 TI - Transfer of plasmids by conjugation in Streptococcus pneumonias. PMID- 6278526 TI - DNase-resistant transfer of chromosomal cat and tet insertions by filter mating in Pneumococcus. PMID- 6278527 TI - A rapid microscale technique for isolation of recombinant plasmid DNA suitable for restriction enzyme analysis. PMID- 6278528 TI - OSHA criteria for laboratory proficiency in blood lead analysis. AB - The OSHA lead standard, 29 CFR 1910.1025, was established to protect the health of workers exposed to the hazards of lead. The standard lists specific requirements to ensure that blood lead analyses--critical indicators of workers at risk--be performed reliably by laboratories. Employers must be laboratories that meet OSHA performance criteria in blood lead proficiency testing programs monitored by the Centers for Disease Control and certain States. This proficiency testing requires that, as a minimum, laboratories must report the equivalent of eight out of nine samples within acceptable limits for the most recent three quarters or similar period. For compliance purposes, OSHA circulates to its staff a "List of Laboratories Approved for Blood Lead Analysis" each quarter. PMID- 6278529 TI - Radiolytic pathways in gamma-irradiated DNA: influence of chemical and conformational factors. PMID- 6278530 TI - The reaction of the hydroxyl radical with glutathione in neutral and alkaline aqueous solution. PMID- 6278531 TI - Radiation-induced tumors in transplanted ovaries. PMID- 6278532 TI - Effect of heat shock on poly(ADP-ribose) synthetase and DNA repair in Drosophila cells. PMID- 6278533 TI - [Role of untwisting proteins in the repair of radiation damage to DNA]. PMID- 6278534 TI - [cAMP-dependent phosphorylation of proteins in irradiated mouse tissues]. PMID- 6278536 TI - Human umbilical blood vessel converts all cis-5, 8, 11, 14, 17 eicosapentaenoic acid to prostaglandin I3. AB - All cis-5, 8, 11, 14, 17 eicosapentaenoic acid (EPA) is presently being evaluated for dietary prophylactic use in thrombo-embolic disorders. EPA inhibits the production of TXA2 and platelet aggregation. We here present results demonstrating that human umbilical arteries convert 14c-EPA to a substance that in aqueous solutions decomposes to 14C-delta 17-6-keto-PGF1 alpha . The conversion rate in rat aortic tissue was found substantially lower. These results in combination with earlier data indicating that EPA does not influence the conversion of arachidonic acid (AA) into PGI2 in human vascular tissue, encourage further research along the lines initiated by the findings of high EPA/AA ratio and low incidence of myocardial infarction in Greenland Eskimos. PMID- 6278535 TI - Radiation-induced osteochondromas. AB - Radiation-induced osteochondromas, either single or multiple, occur more commonly than is generally recognized. The incidence following irradiation for childhood malignancy is approximately 12%. Any open epiphysis is vulnerable. Age at irradiation, time of appearance following therapy, dose and type of radiation, and clinical course in 14 cases are discussed. Due to growth of the lesion and/or pain, 3 tumors were excised. None revealed malignant degeneration. PMID- 6278537 TI - Metabolism of PGI2 by 15-hydroxyprostaglandin-dehydrogenase and delta 13 reductase in different vascular beds of the cat in vivo. PMID- 6278538 TI - Effect of corticosterone, hydrocortisone and prostaglandin fatty acid precursors on the spontaneous motility of the uterus isolated from ovariectomized rats. AB - The effect of corticosterone (CC, 0.4 and 1 microgram/ml) and of hydrocortisone (HC, 20 and 40 micrograms/ml) on the spontaneous motility and on prostaglandin (PG) generation in the uterus from ovariectomized rats, was studied. Both concentrations of CC depressed significantly the frequency of contractions but the isometric developed tension was affected only by the higher dose. HC significantly inhibited the isometric developed tension at both concentrations whereas the contractile frequency was only depressed by the higher one. The CC inhibited motility was accompanied by a reduction in the amount of PGs released from the uterus into the bath solution. In addition, the influences of arachidonic acid (AA), linoleic acid (LA) and gamma-linolenic acid (alpha-LA) - 1 or 2 micrograms/ml - on the depression evoked by CC, were also explored. The fatty acids had no effect on the spontaneous uterine motility except in the case of alpha-LA at 1 microgram/ml. alpha-LA completely blocked the CC-evoked reduction of both tension and frequency; AA (1 microgram/ml) elicited a reversion only on frequency whereas LA had no effect at all. This reversion by a fatty acid PG-precursor might indicate that CC is able to diminish substrate availability for PG synthesis in the rat uterus. PMID- 6278539 TI - Effects of dibutyryl cAMP, LHRH, and aromatase inhibitor on simultaneous outputs of prostaglandin F2 alpha, and 13,14-dihydro-15-keto-prostaglandin F2 alpha by term placental explants. AB - Explants from term human placentas were maintained in culture with daily changes of medium. Daily output of PGF2 alpha and PGFM1 decreased during the course of the incubation. Addition of 4 micrograms/ml DHEAS or 67 micrograms/ml LDL cholesterol had no effect on output of PGF2 alpha or PGFM. Addition of 1.6, 3.2, or 6.4 micrograms/ml of LHRH to the culture plates had no effect on output of PGFM or PGF2 alpha, but LHRH increased hCG output. Dibutyryl cAMP (1mM, 2mM, and 4 mM) increased output of PGF2 alpha, PGFM, and hCG. Aromatase inhibitor decreased hCG output, but it was without effect on output of PGF2 alpha, or PGFM, Significant correlations were demonstrated between progesterone, PGFM, PGF2 alpha, and hCG, suggesting that PGF2 alpha originates in the syncytiotrophoblast cell. The ability of LHRH to stimulate output of hCG but not PGF2 alpha while dbcAMP stimulated both suggests that either PGF2 alpha and hCG arise in different cells or that LHRH does not act through cAMP. PMID- 6278540 TI - Prostaglandin hyperalgesia, V: a peripheral analgesic receptor for opiates. AB - Prostaglandin E2 injected in the rat paw causes hyperalgesia which is antagonized by local injections of opiate and opiate antagonists. In the present investigation in rats it is shown that naloxone has an analgesic effect at doses as low as 2 micrograms/site, injected into the rat hind paw. At a dose that has no analgesic effect (1 microgram/site) naloxone antagonized the analgesia produced by either local or systemic administration of morphine. Local administration of levorphanol (50 micrograms/site) caused a 50% reduction in the intensity of the hyperalgesia induced by prostaglandin E2. A dose four times greater of its isomer, dextrorphan, had little analgesic effect. The present results support the suggestion that this peripheral analgesia is the result of an action of opiates in receptors located at the nociceptors. PMID- 6278542 TI - Angiotensin II receptors are reduced in the CNS of the young Brattleboro rat. AB - In the rat, angiotensin II (AII), following specific interaction with sensitive central nervous system (CNS) receptors promotes release of vasopressin (ADH). We have examined the integrity of this chain of events by comparing the concentration, Bmax, and dissociation constant, Kd, in the CNS of Brattleboro rats (BB), a strain incapable of synthesizing ADH, with Long Evans (LE) control rats that can synthesize ADH. AII binding properties in the hypothalamic-thalamic septal-midbrain (HTSM) area from young and old BB and LE, as well as systolic blood pressure, were determined. There was a reduction in the HTSM-AII receptor concentration of young BB when compared with young LE rats. Young BB had lower pressure than age and sex matched LE controls. Neither Bmax nor pressure was significantly different between older BB and LE. A decline in HTSM-AII receptor concentration with age observed with LE is consistent with observations in SHR and WKY rats. Parallel Scatchard plots obtained indicated the presence of a single class of CNS AII receptors. These data suggest that ADH synthesis and AII receptor concentration are partially interdependent and that the CNS AII-ADH system is redundant in the maintenance of blood pressure. PMID- 6278541 TI - Postsynaptic inhibitory effects of Met- and Leu-enkephalin on endocrine and adjacent neurones in the preoptic-septal region of the guinea pig. AB - Iontophoretic application of enkephalins induced inhibitory effects on unit activity of endocrine and adjacent neurones in the preoptic-septal region. Antagonism or lack of antagonism of these effects by naloxone indicated an action of enkephalins through different opiate receptors. Inhibitory effects of enkephalins were obtained during iontophoretic application of Mg2+, showing that these opioid peptides acted postsynaptically. Because enkephalin acts on endocrine neurones, these opioid peptides might be involved in the control of gonadotrophic hormone release by acting on cell bodies of LH-RH neurones. PMID- 6278543 TI - Pharmacological actions of aminopyridines and related compounds. PMID- 6278544 TI - [Oxygen transport and 2,3-DPG. Physiology and Physiopathology]. PMID- 6278545 TI - [Ectopic ACTH secretion syndrome in a sigmoid neoplasia: an infrequent association]. PMID- 6278546 TI - [Lung carcinoma with inappropriate ADH secretion and Eaton-Lambert syndrome]. PMID- 6278547 TI - [Genetic structure of sedentary cattle of the northern Ivory Coast. Future prospects as a function of crossbreeding of zebu stock]. PMID- 6278548 TI - [Purposes and methodological aspects of a computerized individual monitoring system applied to villagers' cattle of the northern Ivory Coast]. PMID- 6278549 TI - [Central nervous system lesions in cytomegalic disease]. PMID- 6278550 TI - [Health education and group work during in-patient rehabilitation following myocardial infarction (author's transl)]. AB - Starting from the usual in-patient rehabilitation measures for myocardial infarction patients and based on the authors' clinical experiences, a working model is presented concerning the health educational care of patients undergoing rehabilitation procedures immediately following infarction or bypass operation. The new concept is based on a psychosomatic view of the cardiovascular diseases, and uses group-dynamic elements in its methodological approach. The rehabilitational tasks of health education are understood as therapeutic strategies and are aimed at achieving changes on the attitudinal and behavioural level. PMID- 6278551 TI - Epinephrine induced myocardial necrosis: effects of aminophylline and adrenergic blockade. AB - The pathogenesis of myocardial necrosis produced in the albino rat by a single large dose of the potent alpha and beta adrenergic agonist epinephrine was investigated. In confirmation and extension of earlier observations with the alpha adrenergic antagonist tolazoline, it was found that alpha adrenergic blockade with phenoxybenzamine or beta adrenergic blockade with propranolol only partially attenuated the cardiotoxic effect of epinephrine while complete prevention of myocardial injury was achieved with the combined use of the two antagonists. In the presence of alpha adrenergic blockade alone, phosphodiesterase inhibition (aminophylline) caused a dramatic increase of epinephrine cardiotoxicity, demonstrating the importance of unopposed beta adrenergic activation. These results are consistent with the assumption that, in the rat, a cardiotoxic dose of epinephrine produces powerful alpha adrenergic activation which overshadows the effects of the beta adrenergic component of this catecholamine. It is concluded that in epinephrine induced myocardial necrosis, excessive alpha-adrenergic receptor activation in the cardiovascular system is clearly dominant in the initial phases. However, the contribution of the beta adrenergic component of this catecholamine is of considerable importance for the eventual full development of the injury. PMID- 6278552 TI - Properties of hepatic and pancreatic cyclic AMP phosphodiesterase in the cat. AB - Cyclic adenosine-3',5'-monophosphate phosphodiesterase has been characterized in homogenates of feline pancreas and liver. The cyclic AMP phosphodiesterase of feline liver showed a pH optimum at 7.8-7.9. In contrast, cyclic AMP phosphodiesterase activity of feline pancreas increased at up to pH 8.8 without attaining a peak. Hepatic cyclic AMP phosphodiesterase appeared to possess two different enzyme activities, a high and a low affinity form for cyclic AMP. The Michaelis constant for this enzyme was 0.567 and 85.6 microM, respectively. The maximal velocity of this enzyme was 0.135 and 2.84 nmol cyclic AMP hydrolyzed per mg of protein per min. The presence of multiforms of pancreatic cyclic AMP phosphodiesterase was neither demonstrated nor excluded. The kinetics constant for this enzyme for a one-enzyme model without cooperativity was 5.30 microM, and the maximal velocity was 4.97 nmol cyclic AMP hydrolyzed per mg of protein per min. PMID- 6278553 TI - Evaluation of triple drug chemotherapy with or without right nephrectomy in the treatment of bilateral murine Wilms' tumor. AB - The effect of nephrectomy with or without chemotherapy was evaluated in bilateral implanted Wilms' tumor. There was no significant effect of chemotherapy on survival, primary tumor growth or the incidence of metastases to the lungs demonstrated in this study. Regrowth of tumor at the site of unilateral nephrectomy was prevented by the use of chemotherapy when nephrectomy was performed on day 3. The weight of the left kidney at autopsy of animals treated with nephrectomy only was greater than that of animals who received no treatment following bilateral implantation, suggesting that nephrectomy indirectly may have stimulated tumor growth in the residual kidney. The results suggested that future experimental studies of bilateral Wilms' tumor should include surgery and/or irradiation of the remaining kidney. PMID- 6278554 TI - [Hepatitis caused by cytomegalovirus]. AB - Epidemical, clinical, biochemical, cytological and immunological aspects are studied in ten cases of hepatitis occurred endemically in a chirurgical ward between cancerous operated patients. Three were HBsAg positive. None of the others seven had clinical or serological dates enough to attribute them to A nor B virus hepatitis. The clinical and serological data exclude also Epstein-Barr virus or Toxoplasma aethiology. Nevertheless, the objective results of the study have entity enough to classify the other seven no-A no-B hepatitis cases into the group of the cytomegalovirus hepatitis. The high number of positive titles for antibodies anti-CMV in the staff members of the chirugical ward, some with high rates, allow us to give them a role in the spread of the virus and his transmission to the patients' through a failure with the hygenic conditions necessary to look after the wounds. PMID- 6278555 TI - [Neuropathy in angioimmunoblastic lymphadenopathy (author's transl)]. AB - Four cases of angioimmunoblastic lymphadenopathy associated to peripheral neuropathy are described. The neuropathy was mixed, sensory and motor, more or less extensive, always asymetrical. In two cases, the clinical symptomatology and the clinical course were very peculiar, characterized by sensory disorders of a precise topography, circumscribed and sometimes suspended and by a relapsing and remitting course. In the third case, the neurological signs were acute and rapidly extensive with mandatory respiratory assistance. In this case, death occurred after a few weeks and the exact diagnosis was only attained at post mortem examination. In the fourth case the neuropathy was very painful but the course was slow. In all four cases marked and extensive pain was present prior to the neurological disorders. Electrophysiological abnormalities were a constant feature with a marked slowing down of nerve conduction velocity. CSF was normal at the beginning in one case but was otherwise markedly pathological with an increased number of cells due to a large number of lymphocytes ranging from 6 to 40 cells while protein ranged from 60 to 160 mg per 100 ml. Nerve and muscle biopsies were non specific, i.e. neurogenous muscular atrophy and demyelination, except in case n. 4 where specific angioimmunoblastic lymphadenopathy infiltrates were present both in nerve and muscle. In cases 1 and 3 a non specific lymphohistiocytic infiltrate was present in spinal roots and meninges. Corticotherapy was used and efficient in two cases. These data are compared with a review of the literature. Since 1976, 7 cases of angioimmunoblastic lymphadenopathy associated to peripheral neuropathy have been reported. Clinical, electrophysiological and biological features are similar. Only one case underwent a post mortem examination of the central nervous system: a non specific lymphocytic infiltration in the spinal roots and meninges was mentioned. The role of the dysproteinemia associated with the AIL in the occurrence of the neuropathies is discussed. It remains a possible factor only. The role of a massive specific localization of the pathological process is well established in only one case (case 4). Such localization in lymphomas are not unusual. In 2 of our cases as well as in 2 from the literature the pathological findings are non specific: mild lymphoplasmocytic infiltrates of spinal roots and meninges. Those lesions are similar to neuropathies associated to non metastasizing neoplasms or malignant hemopathies. PMID- 6278556 TI - [Isolated sensory trigeminal neuropathy. Discussion on possible toxic etiology (author's transl)]. PMID- 6278557 TI - [Neurological manifestations of oculodentodigital dysplasia: a case report (author's transl)]. PMID- 6278558 TI - [Coxsackie B virus neuropapillitis (author's transl)]. PMID- 6278559 TI - Cardioactive substances that prolong the open state of sodium channels. PMID- 6278560 TI - [Androblastoma of the ovary]. PMID- 6278561 TI - Tuberculosis chemotherapy in low-prevalence countries,--at present and in future. PMID- 6278562 TI - [Tuberculosis in the industrial environment. Epidemiological aspects and preventive measures]. AB - An analysis is made of the tuberculous endemia in several of the most important industrial branches over the last years, as compared with the endemy outside the industrial environment. Although the global morbidity through tuberculosis in industrial enterprises has declined to approximately one half of the initial values (considering the year 1975 as the basis), it is still above the level in the territory. The gap between the two curves, much wider at the beginning of the interval, has narrowed consistently with time. The factors are discussed, that may determine this relative morbidity, and the necessary measures for their neutralization. The necessity for a close cooperation is stressed, between the enterprise physician and the pneumologist (or phthisiologist) from the territorial polyclinic. PMID- 6278564 TI - [Research on normal values of pulmonary ventilation in young persons aged 20 to 30 years]. PMID- 6278563 TI - [Current aspects of drug resistance of the Koch bacillus in the area of one district]. AB - Some aspects were analyzed, of drug resistance of primary and secondary type in mycobacteria strains from patients with pulmonary tuberculosis in a district area, between 1975 and 1979. The results were compared with those obtained in a similar study performed in 1971--1974. While the primary resistance has persisted at an approximately similar level under the aspects of frequency and incidence in the two periods (11% and 13,6%, and 4,0-- and 4,5% respectively), the secondary resistance has marked a significant decrease (36% and 12,3 per 100 000 inhabitants in 1979). The intensive use in the recent years of rifampicin and etambutol in the therapy of tuberculosis explains the new aspects noted in the structure of the primary and of the secondary drug resistance. Thus, an increase was noted in resistance to rifampicin, which, between 1977 and 1979 was above the values recorded for primary resistance to streptomycin. PMID- 6278565 TI - [Corticotherapy of bronchial asthma, using dosed aerosols]. AB - Corticoid therapy with dosed aerosols is one of the achievements that have been made lately in the treatment of bronchial asthma. In view of establishing the efficiency of this type of treatment two preparations have been used: Auxiloson (isonicotinate of dexamethasone), and Becotide (dipropionate of belcomethasone), in two groups of patients. The favourable results obtained involved over 80% of the patients treated with each of these drugs. The use of corticoid therapy with dosed aerosols in the treatment of bronchial asthma reduces many of the risks of the general therapy with corticoids, and is indicated in the maintenance therapy of patients, between asthma attacks, in the intervals when the risk of such attacks is increased, and especially in the long term treatment of corticoid dependent bronchial asthma. PMID- 6278566 TI - [Notes on 2 cases of pneumonia caused by Klebsiella pneumoniae]. AB - In two cases of bacterial pneumonia occurring in adults which were due to Klebsiella pneumoniae germs. The evolution became chronic with extensive invasion of the pulmonary tissues and destruction of the parenchyma. There was also multiple resistance of the germs to antibiotics as evidenced by sensitivity tests. The severe pleural complications have determined serious difficulties in the performance of the treatment. Confronted with such a condition the only therapeutical attitude consisted in testing the known antibiotics with efficiency in Klebsiella infections. PMID- 6278567 TI - [Current chemotherapy of tuberculosis and its effectiveness in Bulgaria]. PMID- 6278568 TI - [BCG vaccination of young children in maternity hospitals and medical dispensaries]. PMID- 6278570 TI - [Detection or diagnosis by symptoms?]. PMID- 6278569 TI - [Evaluation of secondary chemoresistance of Koch bacillus at the present time]. PMID- 6278572 TI - [Debate on evolutionism]. PMID- 6278571 TI - [200 years since the birth of Laennec]. PMID- 6278573 TI - Herpes simplex virus infections of the nervous system: virus isolation and typing from a case of encephalitis. PMID- 6278574 TI - [Clinical and electromyographic study of hemodialysis patients]. PMID- 6278575 TI - [Absorption and metabolism of labelled vitamin D3 in elderly subjects]. AB - This study involved three adult men and six elderly individuals (3 men and 3 women). Adult subjects served as the control group. All subjects were given one dose of 10 microCi of tritiated vitamin D3 (approximately 15 IU) per os. Blood samples were taken 5 min., 1 h, 2 h, 3 h, 4 h, 6 h, 8 h, 24 h, 48 h, 72 h, 96 h, 120 h, 144 h, 168 h and 192 h after administration of vitamin D3. The concentrations of tritiated vitamin D3 and 25-hydroxyvitamin D3, its principal metabolite, were measured in each sample after extraction and purification by high performance liquid chromatography (HPLC). In addition, two adult subjects and three elderly women subjects were given an identical dose of tritiated vitamin D3 approximately 10 days following the end of the initial experience. Blood samples were collected at the times indicated above and analyzed in identical fashion. The percentage of intestinal absorption of vitamin D3 and the amount of vitamin D3 metabolized to 25-hydroxyvitamin D3 were calculated as were the time constants for vitamin D3 and 25-hydroxyvitamin D3 disappearance curves. Review of these parameters confirmed that significant individual variations exist for intestinal absorption of vitamin D3. Study results did not furnish any evidence of a decrease in 25-hydroxylation of vitamin D3, nor any increase in the rate of disappearance of vitamin D3 or 25-hydroxyvitamin D3 which might explain the very low levels of 25-hydroxyvitamin D3, often found in aged persons. PMID- 6278576 TI - Conditions for humoral alpha-adrenoceptor stimulation of renin release in anaesthetized dogs. AB - To examine whether an alpha-adrenergic agonist, methoxamine, influences renin release solely by its haemodynamic effect, experiments were performed in anaesthetized dogs with denervated kidneys. Methoxamine was infused intrarenally at rates which reduced renal blood flow (RBF) by 30-40%. At control blood pressure, renin release rose during infusion of methoxamine from 1.4 +/- 0.7 to 31 +/- 11 microgram/min. A beta-adrenergic stimulator, isoproterenol, did not increase renin release significantly when administered alone into the renal artery, but doubled the effect of methoxamine infusion: at control blood pressure renin release rose from 0.5 +/- 0.3 to 71 +/- 17 microgram/min during combined infusion of isoproterenol and methoxamine. Mechanical constriction of the renal artery left RBF unaltered down to a renal perfusion pressure of 90 +/- 4 mmHg during methoxamine infusion, whereas the lowest autoregulating pressure in control experiments averaged 60 +/- 5 mmHg. At renal infusion pressure below the range of autoregulation, renin release was not further increased by intrarenal infusion of methoxamine. Isoproterenol infusion at low renal perfusion pressure doubled renin release, which was not significantly altered by additional infusion of methoxamine. The stimulatory effect of methoxamine on renin release at control blood pressure could be diminished but not prevented by infusing 2.9% NaCl intravenously in large amounts. These data indicate that methoxamine induces autoregulated dilation of afferent arterioles by disproportionate vasoconstriction on pre-afferent arteries. Thereby afferent arterioles are conditioned for stimulation of renin release by isoproterenol. PMID- 6278577 TI - Conditions for stimulation of renin release by cyclic AMP in anaesthetized dogs. AB - Cyclic AMP (cAMP) is the intracellular mediator of beta-adrenergic stimulation in most tissues. Stimulation of beta-adrenoceptors increases renin release much more at low than at control arterial perfusion pressure. If beta-adrenergic stimulation is mediated by cAMP, this nucleotide should also potentiate renin release at low perfusion pressure. In anaesthetized, propranolol treated dogs, the dibutyryl derivative of cAMP (DB-cAMP), which penetrates cell membranes more readily than cAMP, increased renin release significantly during renal arterial constriction at a perfusion pressure below the range of autoregulation, but no significant effect was observed at control blood pressure. A dose-response relationship could be demonstrated in propranolol treated dogs by administering DB-cAMP at 10, 100 and 1000 micrograms/min at low but not at control blood pressure. Since sodium excretion increased, stimulation of a macula densa mechanism is unlikely, whereas arteriolar dilation, caused by autoregulation at low blood pressure, may condition the juxtaglomerular apparatus for renin release. Infusion of cAMP had no effect on renin release either at control or low blood pressure, whereas 5'AMP exerted a marked inhibitory effect at low blood pressure. We conclude that infusion of DB-cAMP rather than cAMP stimulates renin release at low but not at control blood pressure and that this effect is not mediated by beta-adrenergic receptors; cAMP may be an intracellular mediator of renin release. PMID- 6278578 TI - Application of the law of mass action to design and performance of receptor assays for 1,25(OH)2 vitamin D. AB - The binding of 1,25(OH)2 vitamin D3 to its chick duodenal cytosol receptor protein has been analysed on a law of mass action model. Within the limits of experimental error there is such good agreement, when only one binding site is considered, that it is possible to fit the standard assay curve for practical use. Simple pocket-calculator methods are given for this. The equilibrium constant for the reaction, under the conditions used (pH 7.4, 25 degrees C), was 2.1 X 10(10) 1 X mol-1. A kinetic analysis under the same conditions yielded forward and backward reaction rate constants of 6.5 X 10(6) mol-1 X s-1 X l(2) and 3.1 X 10(-4) s-1 X 1, respectively. The kinetic curve also very closely fitted to the experimental data on the basis of the model. The theory for use of the mass action model to optimize the binding-site concentration to give a minimum absolute error for any measured ligand concentration is presented. The use of the model in assay design, standard curve fitting, quality control and problem solving are discussed. PMID- 6278579 TI - The same type of crystalline inclusions in T-lymphocytes as in plasma cells and B lymphocytes in multiple myeloma. A pathological clone originating from a common lymphocyte stem cell? AB - 2 patients with multiple myeloma had the same type of crystals in plasma cells, B lymphocytes, and T-lymphocytes. It is hypothesized that these crystals are formed in cells belonging to the same clone of pathological cells. The clone would then have to be derived from a common lymphocyte stem cell. PMID- 6278580 TI - Cloning of functional human T lymphocytes by limiting dilution: impact of filler cells and interleukin 2 sources on cloning efficiencies. AB - Conditions for the cloning of alloactivated human lymphocytes by limiting dilution in the presence of interleukin 2 (IL 2) and filler cells have been investigated. Cloning efficiencies were extremely high (at least 40%) when IL 2 derived from pooled peripheral blood mononuclear cells (PBM), stimulated with phytohaemagglutinin in the presence of irradiated Epstein-Barr virus-transformed lymphoid cell lines (LCL), was used in combination with irradiated pooled PBM as filler cells. Cloning efficiencies were reduced almost twofold using autologous filler cells and were further reduced dramatically using autologous IL2, although some clones could still be obtained. In addition, cloning efficiencies were acceptable (over 30%) when IL 2 produced spontaneously from the leukaemic cell Jurkat (M-N) was used. In contrast, sheep erythrocytes and LCL as filler cells failed to allow successful cloning, although LCL were beneficial in the expansion of cloned populations. Other factors being equal, the use of pooled PBM as filler cells and highly active preparations of IL 2 will be the method of choice for cloning functional human T lymphocytes. PMID- 6278582 TI - [Preliminary seroepizootiological studies of the distribution of caprine herpesvirus infections in Switzerland]. PMID- 6278581 TI - The St. Thomas' hospital cardioplegic solution: a characterization in two species. AB - Following detailed investigation and definition of some of the critical factors relating to the composition and use of cardioplegic protective solutions, we have formulated the St. Thomas' Hospital cardioplegic solution number 2. This cardioplegic solution (NaCl 110.0 mM, NaHCO3 10.0 mM, KCl 16.0 mM, MgCl2 16.0 mM, CaCl2 1.2 mM, pH 7.8) is designed for routine clinical use combining optimal protection with simplicity of formulation and administration/infusion. In order to characterize the efficacy of this modified solution, experiments have been carried out in two species: the isolated rat heart and the in-situ dog heart. In parallel protocols, hearts were subjected to ischemic arrest of up to 4 hours. Multidose (every 40 minutes) cardioplegic infusion of the St. Thomas' solution combined with topical hypothermia extended the tolerable period of ischemia from less than 30 minutes to about 120 minutes in the rat and from less than 60 minutes to more than 180 minutes in the dog. These conclusions were based on the measurement of functional indices together with biochemical, cellular chemical and ultrastructural assessments. The studies confirmed the additive protective properties of hypothermia and chemical cardioplegia and the utility of the rat heart model in the assessment of protective interventions. PMID- 6278583 TI - Silicate nephrotoxicity in the experimental animal: the missing factor in analgesic nephropathy. AB - Three silicon-containing compounds (magnesium trisilicate BP, crushed quartz and crushed Arran granite) were added to the drinking water (250 mg/l) of three groups of male guinea pigs for four months. At autopsy all animals receiving magnesium trisilicate showed a focal tubulo-interstitial nephritis mainly affecting the distal nephron. Similar but less intense lesions were encountered in two animals receiving crushed quartz. No renal lesions were found in the control group or in animals receiving crushed Arran granite. The concentration of soluble silicates in drinking water, measured as elemental silicon (Si) by atomic absorption spectroscopy (AAS) was determined for twenty-eight locations in different regions of Britain. Following ingestion of magnesium trisilicate, significant increases in urinary excretion of Si were demonstrated in two healthy adults using atomic absorption spectroscopy. PMID- 6278584 TI - Cirrhosis and primary liver cell carcinoma in Tayside: a five year study. AB - Despite Scotland's well-recognised alcohol problem, there is scant information of the aetiology of cirrhosis in this country. This study of 222 patients, reviewed 197 cases presenting as cirrhosis and 25 cases presenting as primary liver cell carcinoma (PLCC) in the East Tayside area of Scotland between 1975 and 1979. The survey was based on an analysis of all histologically proven cases of cirrhosis and PLCC encountered during a five-year period. There was a constant rate of presentation of cirrhosis of about 40 new patients per year, with a stable pattern of aetiology. About 55 per cent were due to alcohol, and there was no significant change in this proportion over the study. No evidence was found for an increasing female susceptibility or earlier female morbidity in alcoholic cirrhosis. Cryptogenic cirrhosis, cardiac cirrhosis and secondary biliary cirrhosis were more often diagnosed at post mortem. Ninety one per cent of patients with primary biliary cirrhosis were females, but the expected male preponderance in haemochromatosis was not present. In addition to the 25 patients presenting with PLCC, three of the cirrhotic patients developed the tumour by the end of 1979. Seventy one per cent of PLCC cases arose in already cirrhotic livers, none were HBsAg positive. Bronchopneumonia, hepatic failure, gastrointestinal bleeding and cardiac failure were the most frequent causes of death. PMID- 6278586 TI - Different synaptic location of mianserin and imipramine binding sites. AB - The high-affinity binding sites for mianserin and imipramine appear to be locate in different neurons of rat brain. Studies in which lesions were produced with 5,7-dihydroxytryptamine and other studies in which the 5-hydroxytryptamine content was decreased with p-chlorophenylalanine indicate that some of the imipramine binding sites are on serotonin axon terminals and others are on nonserotonergic synapses. The sites that bind mianserin are on postsynaptic serotonin sites as well as on synapses of other neuronal systems. PMID- 6278587 TI - Hyperglycemia of diabetic rats decreased by a glucagon receptor antagonist. AB - The glucagon analog [l-N alpha-trinitrophenylhistidine, 12-homoarginine]-glucagon (THG) was examined for its ability to lower blood glucose concentrations in rats made diabetic with streptozotocin. In vitro, THG is a potent antagonist of glucagon activation of the hepatic adenylate cyclase assay system. Intravenous bolus injections of THG caused rapid decreases (20 to 35 percent) of short duration in blood glucose. Continuous infusion of low concentrations of the inhibitor led to larger sustained decreases in blood glucose (30 to 65 percent). These studies demonstrate that a glucagon receptor antagonist can substantially reduce blood glucose levels in diabetic animals without addition of exogenous insulin. PMID- 6278585 TI - Measurements of benzo(a)pyrene in the city of Bombay for the evaluation of carcinogenic risk. AB - Cigarette smoking, environmental chemicals and ionizing radiations are the three factors known to cause cancer in human beings. The relative importance of each of these is being constantly evaluated. There is an urgent need to monitor the environmental carcinogens on a large scale to assess the role of environmental chemicals in the incidence of cancer in human populations. Polycyclic aromatic hydrocarbons (PAH) are released into the atmosphere as a result of fossil fuel combustion and some of the PAH (e.g. benzo(a)pyrene) are recognised carcinogens. Measurement of benzo(a)pyrene in urban, suburban and rural regions of Bombay is carried out in order to evaluate the possible correlation with lung cancer incidence among different population groups. The variations in the concentration at the three sampling locations are discussed. The wide differences in the concentration at different locations seem to be very suitable for epidemiological investigations. PMID- 6278588 TI - Saxitoxin binding sites in frog-myocardial cytosol. AB - Cytosolic fractions of frog heart homogenates contain large amounts of a soluble, large molecular weight protein that binds the specific neurotoxin saxitoxin with the same high affinity as does the plasma membrane. Another neurotoxin, tetrodotoxin, which ordinarily is competitive with saxitoxin, does not displace saxitoxin from the cytosolic sites or from plasma membrane-enriched vesicular fractions even when its concentration exceeds that of saxitoxin by a factor of 1000. Thus, cytosolic sites are similar to membrane sites in this respect. The vesicular fraction accounts quantitatively for the amount of saxitoxin bound by whole ventricles, so that no appreciable losses seem to occur. Therefore, the cytosolic site probably is a membrane site precursor, although other possibilities cannot be ruled out. In any case, the occurrence of a soluble molecule closely related to the sodium channel provides opportunities for further study of the structure of the sodium channel. PMID- 6278589 TI - The leukotrienes in allergy and inflammation. PMID- 6278590 TI - New theory of hormones proposed. PMID- 6278591 TI - alpha 2-adrenoceptor-mediated hyperpolarization of locus coeruleus neurons: intracellular studies in vivo. AB - Intracellular recordings in vivo from noradrenergic neurons in the rat locus coeruleus showed that membrane potential was hyperpolarized by the administration of clonidine (an alpha 2-adrenoceptor agonist) or after a burst of spikes evoked by intracellular pulses; both types of hyperpolarization were associated with a decrease in membrane input resistance, and both could be blocked by the alpha 2 adrenoceptor antagonist piperoxane. These results suggest that a hyperpolarization of membrane potential mediated by an alpha 2-adrenoceptor underlies both clonidine- and activation-induced inhibition of locus coeruleus cell firing. PMID- 6278592 TI - Oxytocin receptors and human parturition: a dual role for oxytocin in the initiation of labor. AB - The concentration of oxytocin receptors increased in the myometrium of pregnant women and reached maximum levels in early labor. Concentrations of oxytocin receptors were also high in the decidua and reached a maximum at parturition. In vitro, prostaglandin production by the decidua, but not by the myometrium, was increased by the addition of oxytocin. Oxytocin may therefore stimulate uterine contractions by acting both directly on the myometrium and indirectly on decidual prostaglandin production. Oxytocin receptors are probably crucial for the onset of human labor, and the stimulus for the increase in uterine prostaglandins may be oxytocin originating from the fetus. PMID- 6278593 TI - Long-term synaptic potentiation in the superior cervical ganglion. AB - Brief tetanic stimulation of the preganglionic nerves to the superior cervical ganglion enhances the postganglionic response to single preganglionic stimuli for 1 to 3 hours. This long-term potentiation of transmission through the ganglion is apparently not attributable to a persistent muscarinic action of the preganglionic neurotransmitter, acetylcholine, since neither the magnitude nor the time course of the phenomenon is reduced by atropine. The decay of long-term potentiation can be described by a first-order kinetic process with a mean time constant of 80 minutes. We conclude that long-term potentiation, once considered a unique property of the hippocampus, is in fact a more general feature of synaptic function. This form of synaptic memory may significantly influence information processing and control in other regions of the nervous system, including autonomic ganglia. PMID- 6278594 TI - Peptide and steroid regulation of muscle degeneration in an insect. AB - Two types of cell death occur in the intersegmental muscles of the giant silkmoth Antheraea polyphemus. The first results from a slow atrophy of the fibers, and the second is a rapid, programmed dissolution of the muscle. Both types appear to be mediated by endocrine factors. The slow atrophy is brought about by the decline in the steroid molting hormone 20-hydroxyecdysone and can be prevented with exogenous steroid. The rapid degeneration is triggered by the peptide eclosion hormone, but the sensitivity of the muscle to the peptide depends on the history of exposure of the muscle to 20-hydroxyecdysone. PMID- 6278595 TI - Extracellular potassium ions mediate specific neuronal interaction. AB - The giant interneurons from the nerve system of the cockroach Periplaneta americana exhibit a peculiar reciprocal synaptic interaction. The synaptic potentials are not blocked by addition of 5 millimolar cobalt chloride and have an extrapolated reversal potential close to 0 millivolt. Hyperpolarizing current injected into one cell does not spread to the other. Intracellular injection of tetraethylammonium ions into one giant interneuron increases the duration of the action potential of the injected cell to 30 milliseconds and reduces the rise time and amplitude of the postsynaptic response recorded in the other giant interneuron. These results indicate that the interaction between the interneurons is not mediated by conventional chemical or electrotonic synapses.. All evidence points to generation of the potentials by localized increases in extracellular potassium concentrations as a consequence of firing of one neuron. PMID- 6278596 TI - Isolation of infectious bovine rhinotracheitis virus from the soft-shelled tick, Ornithodoros coriaceus. AB - Infectious bovine rhinotracheitis virus was isolated from the soft-shelled tick (Ornithodoros coriaceus). Serological identification of the isolated viruses was confirmed by restriction endonuclease digestion of purified virus DNA. These isolations indicate that the soft-shelled tick may be a vector for infectious bovine rhinotracheitis virus. This may be the first reported isolation of mammalian herpesvirus from an arthropod vector. PMID- 6278598 TI - [Evaluation of protein-losing enteropathy in the cirrhotic patient by assessing the intestinal clearance of alpha 1 antitrypsin (author's transl)]. AB - The purpose of this study was to evaluate protein-losing enteropathy in cirrhotic patients by assaying the intestinal clearance of alpha 1 antitrypsin (C alpha 1), 28 patients were divided into 5 groups: group A = 4 controls, group B = 6 patients with peptic ulcer, group C = 5 cirrhotic patients without esophageal varices (EV), group D = 6 cirrhotic patients with EV but without ascites, group E = 7 cirrhotic patients with EV and ascites. A significant increase in C alpha 1 was found only in groups D and E. The C 1 level was equal to the mean level found in controls multiplied by 2.5 and added to 2 standard deviations. Increases in C alpha 1 levels were not statistically significant when ascites or gastrointestinal involvement were present. C alpha 1 was significantly correlated to the size of EV (p greater than 0.05) and with the Child ratio (p greater than 0.01). No significant correlation was found between C alpha 1 and serum albumin or prothrombin. PMID- 6278597 TI - Interaction of brain synaptic vesicles induced by endogenous Ca2+ -dependent phospholipase A2. AB - Endogenous phospholipase A2 activity of brain synaptic vesicles was Ca2+ dependent and was increased by prostaglandin F2 alpha, calmodulin, adenosine 3', 5' -monophosphate, and adenosine triphosphate, whereas the activity was inhibited by prostaglandin E2 in the absence or presence of calmodulin. Light-scattering measurements demonstrated that stimulation of the enzyme's activity correlated with the induction of vesicle-vesicle aggregation. The effects of these compounds on endogenous synaptic vesicle phospholipase A2 activity may imply a common end point of their purported neuromodulatory actions, and indicate that synaptic vesicle phospholipase A2 may play a central role in presynaptic neurotransmission. PMID- 6278600 TI - [The immediate and remote results of surgical treatment of renal adenocarcinomas. Discussion of two hundred and twenty-two cases (author's transl)]. AB - A series of 222 cases of adenocarcinoma of the kidney are discussed. Many of these cases involved extended carcinomas (stages III and IV, 65 p. cent; stage IV, 41 p cent). The surgical procedures used included 204 extended nephrectomies, 4 partial nephrectomies and 14 laparotomies for the exploration of unextirpable cancers. The mortality rate was 4.5 p. cent and the morbidity rate, 22 p. cent. 168 patients were followed up over a long period after an extended nephrectomy (8 died postoperatively and contact was lost with 28 patients after their operations). The 5-year survival rate was 71 p. cent for stage I, 77 p. cent for stage II, 48 p. cent for stage III and 23 p. cent for stage IV. PMID- 6278599 TI - [Hepatic amebiasis: recent aspects. One hundred and sixty one case reports (author's transl)]. AB - The authors describe their experience of 16l patients (151 adults and 10 children) with hepatic amebiasis. Diagnosis and treatment were carried out at the Treichville Hospital University Center in Abidjan over a three-year period, from September 1976 to November 1979. Diagnosis was established without question in all cases. The indirect immunofluorescence reaction was particularly helpful. All patients were treated by metronidazole or it's derivatives. Semiology, course and therapy are described in detail. A comparison is made with cases published in the literature during the last 15 years. Most of the classical pictures were encountered in this study. The clinical picture may be misleading, for instance when it is suggestive of cancer or when jaundice is present. Immunology is particularly interesting in atypical forms. The authors insist on the unvarying and high morbidity due to this disease which accounts for 2% of all hospitalizations. Prognosis has improved, the mortality rate among adults was 4.5%. A fall in the number of surgical procedures was recorded. Only 10% of the patients underwent surgery. The therapeutic principles advocated by the authors are described. They can be used even when echotomography is not available, as is usually the case in regions where the disease is endemic. PMID- 6278601 TI - [Pulmonary fibrosis in ankylosing spondylitis. Five new cases (author's transl)]. AB - The most common thoracic finding in ankylosing spondylitis (AS) is ankylosis of the costovertebral joints with severe limitation of chest expansion inducing a restrictive syndrome pleural and pulmonary fibrosis can occur during the course of the disease. One-hundred record of patients with AS were reviewed. Five had pleuro-pulmonary involvement. This always included pachypleuritis, usually apical. Roentgenographic pulmonary changes were infiltrates or fibrobullous lesions. Pulmonary function tests showed restriction in four cases, obstruction in one case, and a decreased diffusing capacity in all five cases. PMID- 6278602 TI - [Anatomo-clinical correlations in Berger's disease: evaluation of the prognosis by analysis of one hundred and one cases (author's transl)]. AB - In order to evaluate the prognosis of IgA nephropathy semi-quantitative analysis of tubulo-interstitial and vascular alterations was performed. Three groups of patients are described. In the first group the prognosis has remained favourable until the present time. The prognosis is variable in the second group. In the last group the prognosis is always unfavourable. However, the limited duration of the study may elicit certain reservations concerning the long-term prognosis for the first and second groups. PMID- 6278603 TI - [Factors of early prognosis in medically treated haemorrhages of upper gastrointestinal tract in cirrhotic patients. Statistical study (author's transl)]. AB - In order to define immediate criteria of prognosis for patients with cirrhosis, hospitalized in emergency for upper gastrointestinal tract haemorrhage, 164 cases of haemorrhages medically treated were reviewed. A prognostic study of each variant points to the clinical, endoscopic and biological data connected with death. That study illustrates the prognostic value of traditional data. Patients who died within the first 30 days had a significantly higher incidence of hepatic encephalopathy, hematemesis, ascites, low blood pressure, bleeding oesophageal varices during endoscopy, hypoprothrombinemia, hyperbilirubinemia. Those who survived had a significantly higher incidence of aspirin intake, numerous haemorrhages, good state of health, non bleeding peptic ulcer. Several series of multivariant analysis: multiple regression and linear discrimination, differing from one another in variates were done. They have allowed identification of 11 variables used in establishing a prognostic index. In the series studied, it gives an 87% probability of correct differentiation. Such an index allows an objective estimate of the value and limits of a group of patients in which the efficiency of other treatments could be tested in controlled trials. PMID- 6278604 TI - [Peritoneal dialysis for eliminating copper in patients with Wilson's disease (author's transl)]. AB - We report a case of Wilson's disease, with a rapidly progressive course, in which peritoneal dialysis was performed. The physiopathology of acute features occurring during the course of the disease is reviewed. The different methods for eliminating copper, particularly peritoneal dialysis, are discussed. The authors recall the precautions which are necessary when initiating D-penicillamine therapy. PMID- 6278605 TI - [Sarcoidosis and hyperthyroidism (author's transl)]. AB - The authors report the case of a 31-year-old man with thyrotoxicosis which occurred four months after the discontinuation of long-term corticosteroid therapy for sarcoidosis. Clinical and biological features of thyrotoxicosis were reversible when corticosteroid therapy was reinstated. A review of 27 cases from the literature provides the opportunity to recall the characteristic features of hyperthyroidism associated with sarcoidosis: female prevalence, usual presence of sarcoidosis at onset, secondary occurrence of hyperthyroidism, necessity for simultaneous treatment of both diseases, since corticosteroid therapy alone is infrequently sufficient. PMID- 6278606 TI - [Paralysis of the femoral nerve complicating ilio-psoas hemorrhage after iliac bone transplantation (author's transl)]. AB - The author reported an unusual complication of iliac bone transplantation for grafting of a tibial pseudarthrosis. In a patient having anticoagulant therapy, a large iliac haematoma developed in the donor site and extended deep to the iliacus muscle and through the osteomuscular gap into the retroperitoneal space. Moreover it spread downwards and entrapped the femoral nerve as it lies behind the iliac fascia, above the inguinal ligament. Both a paralytic ileus and a femoral nerve injury commanded surgical exploration through an oblique iliac approach; emptying of the clotted haematoma, section of the inguinal ligament and liberation of the femoral nerve enable to avoid definitive sequelae to the quadriceps but the time required is varying: three years after the accident, recovery is not complete in the operated patient probably owing to delayed surgery (three weeks). PMID- 6278607 TI - [The anatomical forms of mitral incompetency (author's transl)]. AB - A change in any of the parts of the mitral valve apparatus may cause systolic leakage: the mitral ring, the large and small valve, the papillary muscles and the chordae tendinae, hence the possibility of a large number of anatomical and physiological mechanisms and many possible etiologies in the onset of mitral incompetency. For each of the four anatomical constituents of the mitral valve these mechanisms are successively reviewed and their etiologies considered, as various as the sequelae of rheumatic or infective endocarditis, metamorphic calcifications, mucoid degeneration, ischemic or other myocardial changes, constitutional abnormalities. Each type of mitral incompetency has a variable effect on the heart cavities and the pulmonary circulation. PMID- 6278608 TI - [Management of transient ischemic cerebral attacks (author's transl)]. AB - Practitioners should be familiar with the description of transient ischemic attacks (TIA) as 40% are followed more or less rapidly by a stroke. Analysis of the natural history of TIA shows that patients die twice as often from cardiac manifestations of atheroma than from cerebral lesions. This demonstrates the significance of diagnosis and treatment of risk factors. Patients are at increased risk of stroke during the six to twelve months following TIA. Anticoagulants and inhibitors of platelet aggregation seem to reduce this risk. Carotid surgery is indicated only for localized vascular lesions; in other cases surgery increases morbidity. Doppler examination is the best means of detecting cases which need emergency surgical treatment. PMID- 6278609 TI - [Clinical symptoms of mitral valve prolapse. Treatment with metoprolol (author's transl)]. AB - Twelve patients with symptomatic mitral valve prolapse were given metoprolol. The efficacy of the drug was evaluated on clinical improvement and changes in echocardiograms, exercise testing and 24 h ambulatory electrocardiograms. The efficacy of metoprolol was remarkable on palpitations and cardiac arrhythmias recorded on ambulatory electrocardiograms. PMID- 6278610 TI - [Treatment of severe diabetic ketoacidosis in children. A comparative study of two methods (author's transl)]. AB - Fifteen children with severe diabetic ketoacidosis (pH less than 7,10) were treated according to two protocols. Protocol I used in 7 patients, consisted in a low dose insulin therapy by intravenous and intramuscular injections. Protocol II consisted in a low dose continuous intravenous insulin therapy, with insulin added to the rehydration solution at constant concentration (22 U/l). Rehydration and potassium supplementation were similar in both methods. The fall of glucose concentration, rise in venous pH, fall of diuresis, glycosuria, ketonuria were similar in the two groups. With protocol I three hypoglycemias (greater than 4 mM) and three hypokalemias (greater than 3 mEq/l) did occur. With protocol II no hypoglycemia and only one moderate hypokalemia was observed. Thus, continuous infusion of insulin added to the rehydration solution, including isotonic bicarbonate, isotonic saline, glucose and electrolytes is as effective as discontinuous insulin infusion, but with a lower incidence of complications. In addition, this method has the advantage of being easy and applicable everywhere. PMID- 6278611 TI - [Primary tumors of the renal pelvis and ureter: value of postoperative external radiotherapy (author's transl)]. AB - Twenty-four observations of primary malignant tumors of the renal pelvis and ureter derived from epithelial cells were included in this study. Thirteen patients were treated by radiotherapy and surgery. In eleven patients, surgical excision was the only treatment. Although patients were not randomized there was no significant difference between the two groups concerning the following parameters: age distribution, sex ratio, stage of disease, histological grade, surgical procedure. The morbidity related to external radiotherapy (megavoltage technique, delivering 45 grays in 5 weeks) was low: only one patient developed retroperitoneal fibrosis. No patient was lost to follow-up. Local recurrence occurred in 54.5% of the patients treated by surgery alone, against 15.5% after combined radio-surgical therapy. The overall three-year survival rate was 35% without any significant difference between the two groups. The authors advocate postoperative high-voltage radiotherapy for non-disseminated carcinoma of the renal pelvis and ureter. PMID- 6278612 TI - [Radioimmunoassay of serum digoxin levels. Clinical exploration (author's transl)]. AB - This work undertakes, in a second part, the clinical exploration of 947 serum digoxin levels of 281 hospitalized patients on a cardiology ward. Our results which coincide with those of other researchers, have led us to draw certain practical conclusions: the posology is determined first of all according to kidney function, weight and age of the patient. When the treatment is insufficient or on the other hand, poorly tolerated, a serum digoxin level is performed permitting thus: 1) in the case of ineffective treatment: to be sure of the patient's cooperation, to increase the posology if the serum digoxin level is not in the toxic zone, to discover an eventual pharmacokinetic problem; 2) to establish the responsibility of digitalis (when there are signs of intolerance or of intoxication), in case of arrhythmia, in patients with pacemakers, when associated drugs are capable of causing similar adverse effects; 3) to better manage a digitalis treatment in a high risk patient (unstable renal function, advanced myocardial disease, chronic obstructive disease). PMID- 6278613 TI - [Benign clear cell tumour of the lung (author's transl)]. AB - The benign clear cell tumor of the lung is an uncommon neoplasm. It is often confused, before histological analysis, with metastases. The data of one personal observation are similar to those of 19 other cases of the literature. This tumor which occurs with an equal frequency in adults of the two sexes is often asymptomatic. It is discovered after a systematic x-ray control of the lungs. It is located in the peripheral parenchyma and looks like a round, homogeneous, well delimited opacity. Its evolution is always favourable after surgical excision. The benign clear cell tumour is never identified before pathological study. This neoplasm 1 to 6 cm in diameter consists of a uniform proliferation of clear cells filled up with free of intra-vesicular glycogen. Its cytological and structural features are not the same as those of the other primitive bronchopulmonary tumours having an important clear cell contingent. The histochemical and ultra structural studies are at the origin of three hypotheses; is the benign clear cell tumor an apudoma, a smooth muscular neoplasm or an adenoma of the terminal bronchioles? PMID- 6278614 TI - [Anti-tumorous effect of bromocriptine in prolactin-secreting adenomas. A case report (author's transl)]. AB - Bromocriptine inhibits prolactin release. An increasing number of observations, including the one reported in this paper, provide evidence that bromocriptine can also cause significant reduction in the size of prolactin-secreting adenomas. This effect can now be easily demonstrated by CT scanning. Bromocriptine can be used, not only after surgery has failed, but also in order to facilitate the surgical procedure, thereby increasing the surgical success rate. Some authors suggest that prolactin-secreting adenomas be treated with bromocriptine only. Several questions remain open, for instance, why the response to bromocriptine is unpredictable, and whether prolonged therapy with a dopaminergic drug whose effect is not restrained to hypophyseal cells will have adverse side-effects. PMID- 6278615 TI - [Value of myoglobin radio-immuno-assay in myocardial infarction (author's transl)]. AB - Serum myoglobin radio-immuno-assay was performed in 137 patients with myocardial infarction. Two groups, of 102 and 35 patients respectively, were seen in two different hospitals. Serum myoglobin was increased in 72 patients from the first group, and in 30 patients from the second group. In 10 patients from the first group, during the first 24 hours, the increase in serum myoglobin was isolated, without increase in CPK, LDH or SGOT. In the second group, a high serum myoglobin level with normal CPK was found in five patients. Increased serum myoglobin is not a specific indicator of myocardial infarction. PMID- 6278616 TI - [The advantages of lymphography in malignant melanoma (author's transl)]. AB - The authors report the results of their observations on 27 cases of malignant melanoma having undergone lymphography. They insist on the absence of risk of this examination causing diffusion of the primary tumour and the advantages of lymphography in this particular cases: clinically palpable metastatic adenopathy and high risk malignant melanoma without palpable adenopathy. PMID- 6278617 TI - [Paracetamol in oral solution: effectiveness against fever and tolerance in children with measles (author's transl)]. AB - Paracetamol in oral solution, which has become recently available, was given to thirty-one children with hyperthermia due to measles either in the pre-eruptive or the eruptive stages. Rectal temperatures were recorded at regular intervals over the first six hours following the ingestion of a single dose. Falls in temperature were significant and satisfactory. No adverse side-effects were seen. PMID- 6278618 TI - [Hypereosinophilia in cancer and cirrhosis : utility of total eosinophil count (author's transl)]. AB - A total eosinophil count (TEC) and a white blood cell count (WBCC) were performed simultaneously in 213 patients. Discrepancies were found between TEC and WBCC. WBCC did not allow detection of hypereosinophilia (TEC exceeding 300 eosinophils/microliter) in 45 patients. Patients were classified in three groups : controls (n = 27), benign disease (n = 65), and apparently cured cancer (n = 36). There was no significant difference between total eosinophil counts for these three groups (Wilcoxon non-parametric test). The incidence of hypereosinophilia (TEC exceeding 300) was similar in the three groups (11%, 14% and 6%). Average TEC levels were determined in 20 patients with decompensated hepatic cirrhosis, in 31 patients with active cancer, and in 14 patients with infection. These levels were significantly different from those found in each of the three previous groups. (Wilcoxon test). The incidence of hypereosinophilia (greater than 300/microliter) was significantly higher among patients with cirrhosis (50%) or active cancer (58%). The possible mechanisms of this hypereosinophilia and it's eventual prognostic utility are considered. PMID- 6278619 TI - [An empirical combination of antibiotics used in aplasias during chemotherapy for acute malignant hemopathies (author's transl)]. AB - Continuous infusion of amikacin, cotrimoxazole and carbenicillin was the second empirically established combination of antibiotics used when fever occurred during the induction phase of chemotherapy in sixty-five patients (58 acute myeloid leukemias, 5 acute lymphoid leukemias, 2 non Hodgkin lymphomas). Clinical evidence of infection was available in 25 cases and the infection was bacteriologically documented in 19 cases. Therapy was successful in 57 patients (89%). When infection was clinically or bacteriologically documented tha success rates were 92 and 82% respectively. The average length of treatment was ten days. In 25 patients receiving 2 g of amikacin in continuous infusion, the mean serum concentration was 15,9 micrograms/ml; in 17 patients receiving 3 g, the mean serum concentration was 19,4 micrograms/ml. PMID- 6278620 TI - [Acute adreno-cortical insufficiency. Eight case-reports (author's transl)]. AB - Four female and four male adults, with acute adrenocortical insufficiency were admitted to a medical intensive care unit over a period of five years (1975 1980). Three of them were under treatment for Addison's disease. The clinical picture combines fever with signs of shock and dehydration. Increased pigmentation of the skin and laboratory data suggest the diagnosis, which is established by the serum cortisol assay. The etiology was tuberculosis in four patients. Cortical retraction was found in three others. One case followed interruption of non-substitutive corticotherapy. After 48 hours of adequate therapy, all patients had recovered. PMID- 6278621 TI - [Accidental localized vaccinia. A report of six recent cases (author's transl)]. AB - Vaccination against smallpox should be discontinued in all countries except for individuals with a high risk of exposure (WHO, 1980). Since this vaccination is performed less and less often, one must expect complications to occur, the etiology of which may not be recognized. This course of events leads the authors to point out the difficulties in diagnosis and therapy of localized accidental vaccinia encountered in six patients hospitalized in Brest (1971-1979). Diagnosis is considered if the patient himself, or a person he came in contact with, was recently vaccinated. Diagnosis should always be established by virology. Such accidents can be avoided by a faultless vaccination technique and by giving sufficient information to the inoculated subject or to his relatives. PMID- 6278622 TI - [Inheritance of insulin-dependent diabetes mellitus in relation to the HLA system (author's transl)]. PMID- 6278623 TI - [Embolization of benign non tumoral diseases of the kidney (author's transl)]. AB - The material includes 11 traumatic lesions of the kidney after needle biopsy, one hemangioma of the kidney with arteriovenous fistula and 4 renal exclusions for malignant hypertension. In traumatic lesions of the kidney after needle biopsy, embolization was followed by 90,9 p. cent excellent, permanent results without any marked loss of renal parenchyma and with simple post-operative follow-up. A hemangioma with congenital arteriovenous fistula required complex embolization owing to multiple pedicles and caused devascularisation of part of the lower pole of the kidney. The clinical result was satisfactory, but an arteriographic control carried out as a routine 3 months later after the embolization showed the development of newly formed vessels around the renal pelvis. However, no relapse of the hematuria occurred after a follow-up of 2 years. During the same period, spontaneous disappearance of 3 hemangiomas of the kidney sent up for embolization was noted, which led us to treat only hemangiomas which bled and prefer embolization in order to keep the kidney intact. In malignant hypertension, embolization proved efficacious and lasting in one case of small unilateral kidney. Carried out in two stages in one case of chronic nephritis with small kidneys, it proved efficacious on one side, insufficient on the other and had to be followed by nephrectomy. Finally, embolization of a remaining kidney due to chronic glomerulonephritis, technically incomplete, was followed by an uncontrollable attack of hypertension with later death of the patient. The blood supply of the kidney must thus be interrupted completely and embolization should not be undertaken unless it appears technically possible. The degree of pain following embolization suggests the possibility of peridural anesthesia. PMID- 6278624 TI - [Stump carcinoma. A report of ten cases (in a series of 235 gastric cancer cases) (author's transl)]. AB - In a series of 235 gastric cancer cases, 10 cases (4,2%) of cancer of the stump were discovered. Analysis of the results show that the length of time before appearance was always more than 15 years, and 24 years on average. There were no cases of cancer after gastroentero-anastomosis. As concerns pathogenesis, the authors stress the role of bile reflux and of the reduction in stomach acid secretion. They present in details their therapeutic management and insist on the necessity of fiberoptic detection among high risk cases : gastrectomy patients of over 15 years standing. PMID- 6278625 TI - [Congenital fibromatosis. Anatomical and clinical study on three observations (author's transl)]. AB - Three congenital fibromatosis are studied. Six years later, the subcutaneous and skeletal lesions of the first patient have disappeared with no recurrence. Regression was also observed in the second one, a generalized fibromatosis with skeletal, subcutaneous and almost certainly pulmonary and laryngeal lesions. The last case is now progressing (vertebral lesion) with a vascular invasion, the prognostic of which is unknown. A review is made about microscopical appearance, origin and difficulties in diagnosis. PMID- 6278626 TI - [Recent considerations on systemic lupus erythematosus (author's transl)]. AB - Recent advances have shown that the pathogenesis of systemic lupus erythematosus involves a number of interacting factors. Renal involvement is frequent and is one of the leading factor in the prognosis. Renal histopathology is most important in the management of patients. Immunologic tests may help in the diagnosis and also in the evaluation of diseases activity. Although survival has improved dramatically with the use of corticosteroids, there is as yet no evidence that immunosuppressive drugs may be beneficial. PMID- 6278627 TI - [Serum-ferritin assay in patients over seventy. A study on fifty-eight non anemic subjects and thirty-five anemic patients (author's transl)]. AB - An immuno-enzymatic assay of serum ferritin was performed in subjects over 70 years of age. Wide variations in serum ferritin levels were demonstrated in the non-anemic group (17-510 ng/ml). This makes interpretation of results very difficult. In anemic patients a negative correlation was found between serum ferritin levels and TIBC (Total Iron Binding Capacity). When TIBC is under 42 mumol/l or above 63 mumol/l the two values are so closely correlated that serum ferritin assay does not provide any additional information for the diagnosis of anemia. On the contrary when TIBC is between 42 and 63 mumol/l serum ferritin can allow an estimation of tissue iron storage particularly in absolute iron deficiencies. PMID- 6278628 TI - [Value of anti hemolysin and anti-gammahemolysin assays for the diagnosis of staphylococcal infections, particularly of osteoarthritis (author's transl)]. AB - The authors report the results of more than twelve years' personal research on the value of anti-alphahemolysin (AASTL) and antigammahemolysin (AGSTL) assays for the diagnosis of staphylococcal infections, and particularly of osteoarthritis. Among 574 controls, AASTL levels exceeded 2 IU in only 14 subjects (11 of these, levels were between 2 and 3 IU). Levels less than, or equal to, 2 IU were therefore considered normal. This is consistent with previously published data. In 144 staphylococcal infections, confirmed by bacteriology, an increase in AASTL was found in 95 of all cases (65.9%) and in 54 of the 76 osteoarthritis' (71%). Similarly, AGSTL titres, which were under 1/160 (upper normal limit) in 138 controls, were increased in 35 patients with unequivocal staphylococcal infections (61.4%), and in 25 of 36 patients with osteoarthritis (69.4%). These results show that AASTL assay is reliable and often abnormal. However, assay of both hemolysins yields even better results. This dual assay was performed in 57 patients with staphylococcal infection. One hemolysin at least was increased in 47 patients (82.4%). This represents additional positivity in 15.7% of patients when compared to AASTL assay alone, and in 21% when compared to AGSTL assay alone. The high level of positive results with dual assay is even more striking when only staphylococcal osteoarthritis is considered: one or both hemolysins were increased in 91.1% of these patients (31/34). PMID- 6278629 TI - [A survey of 223 salmonella infections diagnosed in the Brest Hospital University Center over a six year period (author's transl)]. PMID- 6278631 TI - [Clinical and pathological study of a peculiar form of tuberculous meningitis: diagnostic and therapeutic problems (author's transl)]. AB - The authors report a case of tuberculous meningitis in which multiple bacteriological samples were negative during the eleven months of the clinical course. There were no other visceral localizations. The fatal outcome was due to vascular lesions (multiple cerebral infarcts), and to CSF flow disorders. The findings of cranial computerized axial tomography and of the pathological examination are reported. PMID- 6278630 TI - [Legionnaire disease and creatinine phosphokinase (author's transl)]. AB - The authors report a case of Legionnaire disease where myolysis was suspected because of increased activities of creatine phosphokinase and it's isoenzymes MBCPK. Significance of MBCPK increase is discussed. Enzyme activity levels are of prognostic significance and should be monitored during the course of the disease. PMID- 6278632 TI - [Congenital toxoplasmosis with hydranencephaly. A case report (author's transl)]. AB - Hydranencephaly is an uncommon finding in congenital toxoplasmosis. The authors report a new case and emphasize the toxoplasmic etiology of this malformation, the possibility of normal neurologic examination at birth, and the usefulness of cranial transillumination whenever congenital toxoplasmosis is suspected in a neonate. PMID- 6278633 TI - [Stevens-Johnson syndrome and pulmonary infection by Mycoplasma pneumonia (author's transl)]. AB - The case-report of an eleven-year-old boy presenting with the association of Stevens-Johnson syndrome and pulmonary symptoms is studied. The course of the disease covered two distinct periods since a one-month interval occurred between the two cutaneo-mucous and pulmonary episodes. Mycoplasma Pneumoniae infection was confirmed by positive seroconversion. The child recovered without sequelae. Indirect immunofluorescence transitorily showed anticytoplasmic antibodies directed against basal epidermic cells. These antibodies were not fixed on the lesions as shown by direct immunofluorescence. The possible connexion between Stevens-Johnson syndrome, Mycoplasma Pneumoniae infection and anticytoplasmic antibodies is under discussion. PMID- 6278634 TI - [Acute infectious myocarditis (author's transl)]. AB - Among the primary myocardiopathies resulting from a known cause, acute infectious myocarditis plays an important part. This condition, usually interstitial myocarditis of viral etiology, occurs predominantly in the infant under two years of age. They are characterized by a hypotrophic hypokinetic myocardiopathy which is very hard to differentiate from endocardial fibroelastosis. Fibroelastosis is probably only one possible result of the course of certain myocarditis'. However, on the whole, prognosis is good and the children recover. Typhoid fever and diphtheria may also compromise myocardial function. PMID- 6278635 TI - [Skin-T lymphocyte-thymus interactions: a new approach in cutaneous immunopathology (author's transl)]. PMID- 6278636 TI - [Echocardiography in the diagnosis of diseases of the pericardium (author's transl)]. AB - Echocardiography is a reliable, non-invasive and inexpensive tool for initial diagnosis and for evaluating the course of various cardiopathies. It was first used in diagnosis of pericardial effusion which is still one of it's best indications. Demonstration of an echo-free area behind the posterior wall of the left ventricle, bordered by dense echoes from the pericardium and remaining visible when the transducer is lowered, lead to the diagnosis of pericardial effusion and to quantification of the amount of fluid. The secondary effects can be assessed by analysis of parietal and valvular motion. PMID- 6278637 TI - [Value of neonatal screening for cystic fibrosis. Evaluation of a neonatal screening program including 34,522 neonates (author's transl)]. AB - Screening for cystic fibrosis in 34,522 neonates was done by assaying proteolytic activity in feces samples spread on special filter paper. Infants were considered at high risk for cystic fibrosis if proteolytic activity was significantly decreased and albumin was found in all the fresh stool specimens. Cystic fibrosis was detected in eight infants, most of whom were already hospitalized for respiratory and/or digestive manifestations suggestive of the disease. Six other patients, one of whom was three-and-a-half years-old, were detected after referral by pediatricians. Two false-negative results were recorded, in infants without detectable pancreatic involvement. Given these results, the authors believe that routine neonatal screening for cystic fibrosis is unnecessary. The various etiopathogenetic mechanisms and the possibilities for investigating patients with cystic fibrosis are discussed. PMID- 6278638 TI - [Acute leukemia associated with breast cancer (author's transl)]. AB - Two cases of acute leukemia in patients with breast cancer are reported. In the first patient, erythroleukemia occurred three years after breast cancer was treated by mastectomy, followed by local radiotherapy ; complementary chemotherapy (melphalan) has been given for twenty-six months. The second patient had onset of acute lymphoblastic leukemia four months after breast cancer was treated by surgery only. This patient subsequently has complete remission. Both patients died shortly after onset of leukemia. In the first patient, bone marrow cytogenetic studies evidenced major abnormalities at an early stage of the disease, with abnormal mitoses in all the cells, whereas, in the second patient, only minor abnormalities were found. A review of previously published cases of breast cancer with acute leukemia was done. Our findings suggest that the association of leukemia with breast cancer may result from therapy is some cases (secondary induced acute leukemias) while in others it may occur spontaneously. PMID- 6278639 TI - [Psoriatic arthritis with renal amyloidosis. A case-report. Review of the literature (author's transl)]. AB - A case of severe psoriatic arthritis with both spinal and peripheral joint involvement is reported. The patient, an HLA B27-positive man, was thirty-one years old at onset. Both antiinflammatory drugs and immunosuppressive agents (chlorambucil followed by azathioprine) were ineffective and the patient became bed-ridden. Thirty-three months after onset a severe nephrotic syndrome developed. Renal biopsy let to diagnosis of secondary renal amyloidosis. The patient died five months later in spite of chemotherapy with melphalan and prednisone. Autopsy could not be performed. A review of the literature showed twelve other cases of psoriatic arthritis with secondary amyloidosis. Men are affected more often than women. Twelve years was the average interval between onset of joint involvement and diagnosis of amyloidosis. In three instances, however, this interval was less than thirtyeight months. Arthritis often included both spinal and peripheral joint involvement. Amyloidosis was evidenced by a nephrotic syndrome in eight out of twelve cases. Prognosis is poor with a fourteen months average interval between onset of clinical amyloidosis and death. PMID- 6278640 TI - [Hypertrophic pulmonary osteoarthropathy with paraneoplastic secretion of four hormones. Considerations on pathogenesis (author's transl)]. AB - The case of a patient with small-cell carcinoma of the lung, bone marrow metastases, and hypertrophic pulmonary osteoarthropathy is reported. Normal growth hormone serum concentrations contrasted with significant increases in ACTH, beta-MSH, calcitonin, and gastrin. A hormonal etiology has previously been suggested for hypertrophic pulmonary osteoarthropathy. Our findings indicate that the hormone responsible for hypertrophic pulmonary osteoarthropathy may be an APUD polypeptidic substance, that differs from immunoreactive GH but is related to somatomammotropins. PMID- 6278641 TI - [Myocardiopathies secondary to myocardial ischemia (author's transl)]. AB - Myocardial ischemia is very rare in childhood but, as in the adult, it can result in very severe heart failure. The discovery of specific signs on the electrocardiogram provides a diagnostic orientation which can be confirmed by biologic data and radionuclide scans. The causes of the affections are related to the child's age group : severe fetal and neonatal anoxia in the neonate (transient myocardial ischemia syndrome) ; anomalous origin of the left coronary artery in the young infant ; acquired coronary anomalies of Kawasaki's disease in the older infant or the young child. PMID- 6278642 TI - [A study on 333 patients with adenocarcinoma of the endometrium treated by combined surgery and radiation therapy (author's transl)]. AB - Three hundred and thirty-three consecutive patients with adenocarcinoma of the endometrium treated from 1958 to 1978 by combined surgery and radiation therapy or radiation therapy alone were reviewed. According to the FIGO staging system, there were 204 stage I, 40 stage II, 24 stage III and 20 stage IV. 40 cases of recurrences were also included in this study. The main parameters influencing survival were clinical staging, depth of myometrial invasion, histologic differentiation as well as the possibilities of radical surgery in these patients. Analysis of therapeutic results shows the superiority of pre-operative intra-cavitary irradiation followed by total abdominal hysterectomy and bilateral salpingo-oophorectomy with an 80% actuarial survival at five years in stage I. Primary surgery followed by radiation therapy gives inferior results, particularly in stage II. Treatment by radiation therapy alone should be reserved for inoperable patients after careful evaluation and gives a 35% actuarial survival at five years. The incidence of serious therapeutic complications was 3,6 %. The early diagnosis of endometrial carcinoma by means of systematic endo uterine explorations in post-menopausal bleeding and the elaboration of combined multi-modality protocol prior to any treatment should lead to a better cure rate for this cancer of increasing incidence. PMID- 6278643 TI - [Anatamo-pathologic aspects of gold-salt induced cholestasis. Report of two cases with ultrastructural study (author's transl)]. AB - The authors report two cases of gold-salt-induced cholestasis. They make a review of the thirteen cases previously related in the literature ; the cholestasis was constant, the hepatocytic necrosis was unusual and the gold-salts were difficult to visualize in the liver biopsy and more especially at the ultrastructural level. The immunological mechanism of hypersensibility is well established ; the possibility of an associated hepatotoxicity is debated. PMID- 6278645 TI - [Varioliform gastritis revealed by edema with hypoproteinemia (author's transl)]. AB - The authors report the case of a woman in whom edemas with hypoproteinemia, but without digestive symptoms, were present for several months. Roentgenography, fiberoptic endoscopy, and histopathology showed typical varioliform gastritis. Hypoproteinemia was due to plasma protein leakage into the digestive tract. Edema disappeared spontaneously while gastric pain developed. PMID- 6278644 TI - [Granulomatous histiocytosis in a 70-year-old woman, revealed by prolonged fever ; osseous lesions in the femur and tibia were associated with diabetes insipidus and hypernatremia (author's transl)]. AB - The different patterns of histiocytosis X are usually classified into three forms according to their clinical and pathological expression. One form is acute and disseminated ; the two others are chronic, unifocal or multifocal. Borderline cases and intermediate forms can be seen. For instance, onset can be acute or subacute in chronic disseminated forms, such as Hand-Schuller-Christian disease ; acute or subacute exacerbations may occur during the course of the disease. We report an observation of this type which is remarkable for three reasons. Fever was the initial manifestation and remained apparently isolated for one year. Subsequent osseous involvement was limited to the femur and tibia. Hypernatremia developed as a result of both diabetes insipidus and disrupted thirst control. These unusual features of our case-report led us to review previously published data. PMID- 6278646 TI - [Relapse of Addison's disease after apparent recovery : a case-report]. AB - In our observation, antituberculous therapy was given systematically, although neither interview nor bacteriology had provided unequivocal evidence of tuberculosis. Corticosteroid withdrawal by the patient was uneventful. After, withdrawal, the 17 OH steroids and the response to metyrapone were normal. The water load test was normal. After insulin, only cortisol levels did not increase. Salt-loss with hyponatremia then occurred spontaneously ; 17 OH steroids were low and did not increase after corticotrophin. PMID- 6278647 TI - Some important aspects of the palatal pleomorphic adenoma. PMID- 6278648 TI - Charcot-like joints in calcium pyrophosphate dihydrate deposition disease. PMID- 6278649 TI - [Arterial hypertension today in Africa. Milan Symposium]. PMID- 6278651 TI - [Viruses in the feces and their significance in the occurrence of diarrhea]. PMID- 6278650 TI - [Diagnosis of neural amyotrophy]. PMID- 6278652 TI - [Continuity of the treatment of peptic ulcer patients after hospitalization]. PMID- 6278653 TI - [Evaluation of Bordetella strains used in the production of anti-pertussis vaccine in stationary culture]. PMID- 6278654 TI - [Initial studies for the development of processes in the production of bacterial vaccines by agitator culture]. PMID- 6278655 TI - [Development of a fermentation process in the production of pertussis vaccine]. PMID- 6278656 TI - [Comparative study of HBP-2 and FGP-1 culture media in the production of Bordetella pertussis by fermentation]. PMID- 6278657 TI - [Concentration of pertussis vaccine using citric acid]. PMID- 6278659 TI - [Experimental relation between leukocytosis and other factors of Bordetella pertussis]. PMID- 6278658 TI - [Comparative study of the potency of the pertussis component in DPT vaccine produced in stationary culture and by fermentation]. PMID- 6278661 TI - [Biological selection of Bordetella pertussis and Salmonella schottmuelleri using the diabetic mouse]. PMID- 6278660 TI - [Experimental relation between histamine-sensitizing factor and the protective antigen of Bordetella pertussis]. PMID- 6278662 TI - Two histologically different, synchronous carcinomas in a single male breast. A case report. AB - A rare case of male breast carcinoma is reported. The operative specimen revealed two histologically distinct tumours, a papillary carcinoma and a colloid carcinoma. Features may support the theory of multicentric origin of breast cancer. PMID- 6278663 TI - Myofibroblastic stromal reaction in retracted scirrhous carcinoma of the breast. PMID- 6278664 TI - Pituitary Cushing's syndrome and Nelson's syndrome: diagnostic criteria, surgical therapy, and results. AB - Eight patients with pituitary Cushing's syndrome and 2 with Nelson's syndrome were followed from one to ten years after removal of pituitary adenomas. A detailed assessment of the pituitary-adrenal axis was obtained in all patients when last seen, save the first, who had undergone a complete hypophysectomy ten years previously. Long-term observations have shown sustained endocrine cure in 7 of 8 patients with pituitary Cushing's syndrome. One patient with Nelson's syndrome was also cured. There was no operative morbidity or mortality. There were no instances of diabetes insipidus. The long-term results in this study indicate that patients with pituitary Cushing's syndrome have a better than 90% chance of being cured after transsphenoidal removal of the pituitary (micro) adenomas. Current diagnostic and therapeutic concepts in the management of pituitary Cushing's syndrome are discussed in detail. PMID- 6278665 TI - Intramedullary spinal cord metastasis. AB - Intramedullary spinal cord metastases are rarely the presenting manifestation of a previously undiagnosed neoplasms. We report such a case in which a subacutely progressive motor and sensory spinal cord syndrome was the initial problem. The differential diagnosis and the difficulties in distinguishing intramedullary from extramedullary mass lesions are discussed. The special features of tumors of the lung and their predilection for metastasis to the central nervous system are considered. PMID- 6278666 TI - [Blood levels of beta-endorphin and beta-lipotropin in patients with Cushing's syndrome]. PMID- 6278667 TI - [Effect of exercise on the level of cyclic adenosine monophosphate in patients with different variants of neurocirculatory dystonia]. PMID- 6278668 TI - Immunohistochemical localization of cyclic AMP during normal and abnormal chick and mouse limb development. AB - This paper describes the immunohistochemical localization of cAMP during limb chondrogenesis in talpid3 chick, brachypod mouse, and normal embryos. Comparisons were made between chick wing buds at Stages 22, 25, and 30, and mouse hind limb buds at Days 11, 12.5 and 14. At Stage 22, the normal mesenchyme in the chick displayed areas of bright fluorescence compared to a lesser intense and more evenly distributed fluorescence in talpid3. Sections of the central region from normal Stage 25 limb buds exhibited an intense fluorescence that was uniformly distributed, whereas, in talpid3 staining was more mosaic with some areas fluorescing brightly and others showing little fluorescence. At Stage 30 the staining pattern was similar between normal and talpid3, with the fluorescence being brighter in the cartilage tissue than in the surrounding soft tissue. Difference in the staining patterns of normal and brachypod limb tissue were not detectable. At Days 11 and 12.5, tissue from both genotypes displayed a very bright, uniform fluorescence. In the 14-day hind limb buds, the staining patterns were comparable to those observed in Stage 30 chick wing buds. However, under in vitro conditions conducive for the expression of the chondrogenic phenotype, differences in the intensity and extensiveness of fluorescent staining were detectable in cultures derived from 12-day normal and brachypod hind limb mesenchyme. Compared to the control, the uneven distribution of immunofluorescence in the talpid3 limb buds and the differences in intensity and extensiveness of fluorescence in the brachypod cultures support the hypothesis that cAMP is involved in limb cartilage differentiation. PMID- 6278669 TI - Two forms of Ca++-activated neutral protease in platelets. PMID- 6278670 TI - Streptokinase inhibits sympathetic neurotransmission in isolated blood vessels of the dog. PMID- 6278671 TI - The enhancement of streptokinase activation of plasminogen by nonionic detergents and by serum albumin. PMID- 6278673 TI - [Aujeszky's disease in young chicks]. PMID- 6278672 TI - Activated prothrombin complex concentrates: approaches to their preparation. AB - Locally produced prothrombin complex concentrate was activated by different methods and the resultant inhibitor bypassing activity compared with commercially produced activated concentrates using an assay based on the shortening of the clotting time of haemophilic plasma containing inhibitor to factor VIII. Analytical chromatography suggested that for most of these products the inhibitor bypassing activity could be explained in terms of classical activated coagulation factors. In addition, a preparation of Bridge Anticoagulant Neutralising Activity was examined. The results indicate this product merits in vivo investigation as a possible treatment for inhibitor patients. PMID- 6278674 TI - Estimation of B-cells transformed by Epstein-Barr virus in patients with congenital agammaglobulinemia. AB - In vitro immunoglobulin synthesis was measured in lymphocytes from four patients with congenital agammaglobulinemia (cA gamma) stimulated by two different polyclonal B-cell activators, pokeweed mitogen (PWM) and Epstein-Barr virus (EBV). In PWM-stimulated cultures, patient T-cells treated with mitomycin C (MMC) were able to help the immunoglobulin (Ig) synthesis of normal B-cells. Patient B cell-enriched fraction not containing surface Ig positive cells did not produce Ig in combination with MMC-treated autologous or allogeneic T-cells. Patient lymphocytes were infected with EBV and the subsequent production of Ig was measured. In lymphocytes from control subjects, exponential growth of the cells having EBV-associated nuclear antigen (EBNA) was shown to be associated with an exponential increase in Ig secretion within 1 week after EBV infection. However, in lymphocytes from three of the four patients, it took 2, 4 and 10 weeks, respectively, until lymphocyte-transformation and subsequent Ig-secretion were observed. Lymphocytes from one patient were not transformed nor did they secrete Ig after EBV infection. These results may imply that a small number of B-cells are present in peripheral blood of most of patients with cA gamma, and that they are able to produce the Ig after transformation by EBV which takes a much longer time than in controls. PMID- 6278675 TI - Brain angiotensin-converting enzyme activity in experimental hypertensive rats. AB - Angiotensin-converting enzyme (ACE) activity was measured in six areas of the brain of normotensive and experimental hypertensive rats; one-clip, one-kidney (1 c, 1-k) and one-clip, two-kidney (1-c, 2-k) Goldblatt hypertensive (GH) rats. ACE activity was consistently high in the thalamus of normotensive and both 1-c, 1-k and 1-c, 2-k GH rats. However, the enzyme activity in the hypothalamus of 1-c, 2 k GH rats was significantly higher than that of normotensive rats, while there was no significant difference in the enzyme activity between normotensive and 1 c, 1-k GH rats. These results demonstrate that in 1-c, 2-k GH rats, increased ACE activity in the brain may play a central role in the hypertension. PMID- 6278676 TI - Enzymatic activities of liver serine esterases during the reticuloendothelial system phagocytosis blockade by Carbaryl, an anticholinesterasic insecticide. PMID- 6278677 TI - Effects of caramel color (ammonia process) on mammalian vitamin B6 metabolism. PMID- 6278679 TI - Biochemical studies in infective amosite pneumoconiosis. AB - The effects of Candida albicans, of amosite dust, and of the 2 agents combined, on some biochemical parameters of the lung in male guinea pigs were determined. Adult guinea pigs were intratracheally injected with 50 mg of amosite dust and 0.3 mg of viable suspension of mycelia of Candida albicans. The other groups received either dust or organism or none. Biochemical measurements done at 30, 60 and 90 days after infection showed that the superimposed state produced more rapid and drastic changes in pulmonary contents of collagen, mucopolysaccharides and phospholipids and activities of lactic dehydrogenase and succinate dehydrogenase. The levels of lactic acid and pyruvic acid were also more profoundly changed after amosite + Candida. These results indicate that Candida albicans which is commonly found in upper respiratory tract could aggrevate the lesions caused by exposure to asbestos. PMID- 6278678 TI - Immunotoxicity of pentachlorophenol (PCP): increased susceptibility to tumor growth in adult mice fed technical PCP-contaminated diets. PMID- 6278680 TI - [Biochemical disorders of bone involving calcium, phosphorus and phosphatases affecting the jaw]. PMID- 6278681 TI - Failure of brain cytochrome alpha , alpha 3 redox recovery after hypoxic hypotension as determined by in vivo reflectance spectrophotometry. AB - Rats were subjected to a 30 minute period of combined hypoxia (F1o2 = .08) and hemorrhagic hypotension (MAP = 30 mm Hg), then resuscitated by restoration of F1o2 = .30 and reinfusion of shed blood and saline. Intracranial blood volume, hemoglobin saturation, and the cytochrome alpha, alpha 2 redox state were monitored through the intact skull during hypoxic hypotension and after resuscitation utilizing reflectance spectrophotometry. Although resuscitation returned arterial blood pressure, arterial pO2, and hemoglobin saturation toward normal, a sustained, significant (p less than .005) reduction in cytochrome alpha , alpha 2 remained. A parallel series of rats was subjected to identical hypoxic hypotension. At designated intervals the animals were sacrificed to determine brain ATP, ADP, and inorganic phosphate (P1). The data are discussed in terms of relationships between high energy phosphate metabolism and recorded changes in cerebral cytochrome alpha , alpha 2 redox state. PMID- 6278683 TI - [Ultracytochemistry of cyclic AMP metabolism]. AB - The paper is concerned with the role of cyclic nucleotides in physiological and pathological processes of the cell vital activity. Principles of histochemical detection of adenylate cyclase and phosphodiesterase of cAMP are discussed. Electron-histochemical data on ultrastructural localization of these two enzymes are reviewed and practical recommendations are given for their electron histochemical detection. PMID- 6278684 TI - [Effect of the structural integrity of Na,K-ATPase preparations on the ability of lithium to substitute for sodium in enzyme activation]. AB - The ability of Li+ to substitute for Na+ in activating ouabain-sensitive ATPase of the rat kidney was studied on three kinds of the enzyme preparation: a) crude membrane fraction, b) purified enzyme, c) tissue slices. The preparations were free of endogenous Na+ and K+. The ATPase activity of slices was estimated both biochemically and cytochemically. In slices, the extent of Li+--Na+-isomorphism was about 10-fold higher than that in the purified enzyme. It is concluded that the biochemical procedures involved in the enzyme isolation procedure cause the damage of membranes, thus decreasing Li+-affinity to the Na+-dependent site. PMID- 6278682 TI - Nonbacterial nonfungal pneumonia following marrow transplantation in 100 identical twins. AB - Pneumonia attributable to causes other than bacterial or fungal infection is a major complication of allogeneic marrow transplantation. In this study, the incidence, mortality, and possible risk factors for the development of nonbacterial, nonfungal pneumonia after 100 syngeneic marrow transplants for hematological malignancy are reviewed and compared with the results found in 351 allogeneic marrow transplants performed during the same time period. Both the incidence and mortality of pneumonia were far lower among syngeneic patients, especially for pneumonia associated with cytomegalovirus. Idiopathic pneumonia, however, occurred with about equal frequency in the two populations. Among twins, an increased incidence of pneumonia was seen in older patients and in those who received chemotherapy in addition to cyclophosphamide and total body irradiation as part of the preparative regimen for transplantation. PMID- 6278685 TI - Resistance to Moloney murine sarcoma virus (M-MuSV) tumor induction is associated with natural antibody production to "endogenous" Moloney leukemia virus (M-MuLV) in BALB/Mo mice. AB - BALB/Mo mice are characterized by early expression of "endogenized" M-MuLV and are resistant to M-MuSV tumor induction. Furthermore, compared to normal BALB/c mice, sera from BALB/Mo mice exhibit a significant reactivity which is specific for M-MuLV when tested in a 125I-labelled Staphylococcus protein A binding assay. The possible significance of this reactivity in conferring tumor resistance is explored. PMID- 6278686 TI - Histology and cytometrics in human breast cancers assayed for the presence of prolactin receptors. AB - Histology and cytometrics were evaluated in 50 cases of human breast infiltrating carcinomas (44 ductal, 6 lobular) assayed for the presence of prolactin receptors (PR). The tumors were considered PR positive when the level of specific binding reached or exceeded 0.5%. Twenty-six ductal infiltrating carcinomas and 4 lobular ones were PR positive. No strict correlation was found between PR and degree of histologic differentiation (expressed in grades) or menopausal status. Better differentiated (grade I) ductal carcinomas were, however, mostly PR negative. Evaluation of the mean nuclear diameter (MND) and of the maximal epithelial cellularity (MEC) revealed that in ductal carcinomas a high level of PR (greater than 3% specific binding) was significantly correlated with a high MND. It is concluded that, contrary to that observed in estrogen receptor-positive tumors, PR-positive human ductal carcinomas are more likely to have large nuclei, i.e., cytologically anaplastic, whereas better differentiated (grade I) carcinomas are generally PR negative. PMID- 6278687 TI - Combination chemotherapy for metastatic brain tumors. AB - Forty-four patients with central nervous system metastases were treated with combination chemotherapy (adriamycin, VM 26 and CCNU). The best results were obtained in breast adenocarcinoma and small cell lung carcinoma with multiple, small cerebral metastases and without concomitant visceral involvement at other sites. The potential effectiveness of this regimen to prevent cerebral metastases is discussed. PMID- 6278688 TI - Oat cell carcinoma of the esophagus. Report of a case. AB - A case of primary oat cell carcinoma of the esophagus is described. The oat cell pattern appeared intermingled with small areas of squamous carcinoma. The origin of this mixed tumor is discussed together with the literature on esophageal carcinomas with an oat cell pattern. PMID- 6278689 TI - [Myxedema and polyneuropathy]. PMID- 6278690 TI - [Hepatocellular carcinoma 34 years after injection of thorotrast]. PMID- 6278691 TI - [Inguinal herniotomy with silk or Dexon. A randomized comparison with special attention on recurrence rate]. PMID- 6278692 TI - [Diagnosis of bovine coronaviruses using mouse erythrocyte suspensions stabilized with formalin]. AB - Bovine coronavirus isolated from calf faeces diseased with gastroenteritis and passaged to colostrum-free calves agglutinated mouse and rat erythrocytes. The agglutination reaction depended on temperature and took place only at a temperature of 4 degrees C. At a temperature of 37 degrees C the agglutinate broke down within 15 minutes. The coronavirus could be detected by the haemagglutination test in the contents of the small and large intestines and in the faeces of experimentally and naturally infected calves. The agglutination capacity of mouse erythrocytes was not affected by careful fixation of these erythrocytes with formalin and subsequent lyophilization and remained unchanged for as long as 52 weeks of storage at a temperature of 4 degrees C. It was demonstrated by a comparative examination of 182 samples of the faeces of calves suffering from diarrhoea that haemagglutination test was as sensitive as electron microscopy. PMID- 6278693 TI - Swine pox in Nigeria. AB - An outbreak of swine pox on a pig farm in Ibadan, Nigeria is reported. Diagnosis was based on clinical and pathological signs and the observation of poxvirus particles in a serum sample. Neonatal deaths were common in the outbreak. Indigenous pigs did not show any sign of the disease even though mixed breeds were kept in the same pens. It is suggested that indigenous pigs were probably carriers of the swine pox virus. PMID- 6278694 TI - Incidence of rotavirus, adenovirus and herpesvirus infection in pigeons. PMID- 6278695 TI - Immunosuppression in toxoplasmosis: studies in sheep with vaccines for chlamydial abortion and louping-ill virus. AB - Toxoplasma gondii infection was established in sheep seven days (acute infection) or 28 days (chronic infection) before they were given enzootic abortion (EAE) vaccine alone or simultaneously with louping-ill (LI) vaccine. Uninfected controls received the same vaccination schedule. The serological response of acute toxoplasma-infected sheep to initial LI vaccination was significantly depressed but after revaccination four weeks later these sheep demonstrated a response equal to that of the revaccinated controls. Infected sheep also gave a significantly poorer response to EAE vaccine than control animals, the effect being more marked in those with chronic infection. Revaccination evoked no clear response in any group but on three of five sampling days antibody titres of controls were significantly higher than those of infected sheep. PMID- 6278696 TI - Field study of coccidial and rotaviral diarrhoea in unweaned piglets. AB - A study of diarrhoea in unweaned piglets was carried out in nine herds, with special reference to the enteropathogenic agents which could be demonstrated. Coccidial (Isospora suis) and rotaviral infections were both identified, either singly or in combination. More extensive studies of I suis infection were undertaken in two of the herds and it was found that diarrhoea occurred most commonly in five- to 14-day-old piglets. Piglets with I suis infection were not necessarily diarrhoeic but grew poorly compared to uninfected piglets. I suis infection in litters correlated with oocyst excretion in sows. In herds with I suis infection, amprolium and monensin were used in the sow ration to achieve control, and in one herd oral dosing of piglets with amprolium in the first three or four days of life was carried out. PMID- 6278697 TI - Long distance transport of foot-and-mouth disease virus over the sea. AB - The conditions required for the transport of foot-and-mouth disease (FMD) virus in the atmosphere over long distances and in sufficient concentrations to cause infection in exposed animals are described. Using these factors a series of 23 outbreaks of FMD in Europe, where the original outbreaks were separated from later outbreaks by sea passage, have been investigated. The findings obtained support the hypothesis that under certain conditions the airborne transmission of FMD over a long sea passage is possible. PMID- 6278699 TI - Evidence for the presence of pro-gamma-melanotropin, the NH2-terminal fragment of the corticotropin-beta-lipotropin precursor, in corticotropin-producing tumours. AB - Five corticotropin-producing tumours were examined for peptides related to the corticotropin-beta-lipotropin precursor. Two were basophil pituitary adenomas and three were bronchial carcinoids. The cells of the two pituitary adenomas stained with antisera against beta-endorphin and against pro-gamma-melanotropin, the NH2 terminal fragment of the corticotropin-beta-lipotropin precursor, but not with antisera against alpha-melanotropin or beta-lipotropin. The corticotropin-storing tumor cells of the bronchial carcinoids stained with antisera against beta endorphin, beta-lipotropin or pro-gamma-melanotropin. Only one of the three bronchial carcinoids contained cells reacting with the antiserum against alpha melanotropin. Although the two types of corticotropin-storing tumours (pituitary adenoma and bronchial carcinoid) differed with respect to beta-lipotropin content, the over-all picture indicates that the proteolytic processing of the corticotropin precursor proceeds along similar lines in tumour cells and in pituitary corticotrophs. An acetic acid extract of one of the bronchial tumours was subjected to gel chromatography and immunochemical analysis of material related to pro-gamma-melanotropin. The immunoreactive material displayed a considerable size heterogeneity, with the predominant components having a molecular weight larger than that of authentic pro-gamma-melanotropin. PMID- 6278700 TI - Malignant fibrous histiocytoma of the lung. AB - A case of malignant fibrous histiocytoma of the lung is reported. The tumour margin was well circumscribed, showing an expanding border and no capsule. The main part of the tumour was composed of spindle-shaped fibroblast-like cells arranged in broad fascicles with a partially storiform pattern. Other parts of the tumour were arranged in a haphazard pattern, containing many mononucleated and multinucleated giant cells. Ultrastructurally six different cell types were encountered. The dominant type was a fibroblast-like cell; also present were many giant cells and some histiocyte-like cells, together with their intermediate forms, and few undifferentiated mesenchymal cells. We consider this tumour to have developed from the peribronchial connective tissue; it has the same cellular composition as the malignant counterpart arising in soft tissues. PMID- 6278698 TI - Prevalence of rotavirus antibody in chickens and horses in Louisiana, USA. PMID- 6278701 TI - Immunohistochemical detection of carcinoembryonic antigen (CEA) in parotid gland carcinomas. Analysis of 52 cases. AB - The presence of CEA in parotid gland tumours was studied by immunohistochemical methods. 52 cases were analysed. 7 of 8 adenocarcinomas, 3 of 5 cystadenocarcinomas, 3 of 4 adenoid cystic carcinomas and all 3 salivary duct carcinomas were positive for CEA. 5 of 8 squamous cell carcinomas and 9 of 21 carcinomas in a pleomorphic adenoma were also positive for CEA. The anaplastic carcinomas were negative. The distribution pattern of the presence of CEA in the carcinomatous and the adjacent normal or inflamed tissue was analysed. The results are discussed with regard to their histogenetic and diagnostic implications. PMID- 6278702 TI - Lactoferrin and lysozyme in carcinomas of the parotid gland. A comparative immunocytochemical study with the occurrence in normal and inflamed tissue. AB - Lactoferrin and lysozyme, parts of the non-specific defense system, were studied in normal and diseased parotid glands, using the immunohistochemical PAP-method. 31 normal and inflamed glands were investigated. The presence of lactoferrin and lysozyme was demonstrated in the acinar cells and some duct cells. The amount of these substances was increased in obstructive parotitis. The 52 carcinomas showed a distinct distribution pattern for lactoferrin (positive cases: adenocarcinomas 5 of 8; cystadenocarcinoma: 3 of 5; adenoid cystic carcinomas 2 of 4; salivary duct carcinomas 2 of 3). Some of the carcinomas in pleomorphic adenomas were positive for lactoferrin. Squamous cell carcinomas and anaplastic carcinomas were constantly negative. All carcinomas were negative for lysozyme. These observations are discussed with respect to their physiological and pathological significance. PMID- 6278703 TI - The effect of a temperature-sensitive lesion in the alkaline DNase of herpes simplex virus type 2 on the synthesis of viral DNA. PMID- 6278704 TI - Further studies on the inhibition of cellular protein synthesis by vesicular stomatitis virus. PMID- 6278705 TI - Association of 3' terminal RNA sequences with avian leukosis viruses causing a high incidence of osteopetrosis. PMID- 6278706 TI - Translation of foot-and-mouth disease virion RNA and processing of the primary cleavage products in a rabbit reticulocyte lysate. PMID- 6278707 TI - DNA and histone synthesis of butyrate-inhibited BSC-1 cells infected with SV40. PMID- 6278709 TI - Identification of a 39,000-dalton protein in cells transformed by the FBJ murine osteosarcoma virus. PMID- 6278708 TI - The evolution of polyoma-transformed rat cell lines during propagation in vitro. PMID- 6278710 TI - Control of abundance of immediate-early mRNA in herpesvirus (pseudorabies) infected cells. PMID- 6278711 TI - Association of murine leukemia virus gag antigen with extracellular matrices in productively infected mouse cells. PMID- 6278712 TI - Acylation of viral spike glycoproteins: a feature of enveloped RNA viruses. PMID- 6278713 TI - Cores in foot-and-mouth disease virus. PMID- 6278714 TI - Epstein-barr virus-related DNA-binding proteins induced by n-butyrate in P3HR-1 cells. PMID- 6278715 TI - Analyses of the genomes of prototype pichinde arenavirus and a virulent derivative of Pichinde Munchique: evidence for sequence conservation at the 3' termini of their viral RNA species. PMID- 6278716 TI - Cell killing by simian virus 40: evaluation of the role of extracellular calcium. PMID- 6278717 TI - Helper viruses associated with avian acute leukemia viruses inhibit the cellular immune response. PMID- 6278718 TI - Mapping of nonconditional and conditional mutants in the src gene of Prague strain Rous sarcoma virus. PMID- 6278719 TI - The oncogenicity of avian adenoviruses. III. In situ DNA hybridization of tumor line cells localized a large number of a virocellular sequence in few chromosomes. PMID- 6278720 TI - The adsorption and degradation of foot-and-mouth disease virus by isolated BHK-21 cell plasma membranes. PMID- 6278721 TI - Studies on the interaction of HVJ (Sendai Virus) with liposomal membranes. HVJ induced permeability increase of liposomes containing glycophorin. PMID- 6278722 TI - Molecular mechanisms involved in morphological variation of avian sarcoma virus infected rat cells. PMID- 6278723 TI - Activation of herpes simplex virus (HSV) type 1 genome by temperature-sensitive mutants of HSV type 2. AB - Previous studies have demonstrated that herpes simplex viruses (HSV) type 1 (HSV 1) and type 2 (HSV-2) can be maintained in a repressed form in human embryo lung cells. Reducing the incubation temperature or superinfecting with a heterologous herpesvirus, human cytomegalovirus (HCMV), results in activation of virus replication. We now report that superinfection with a partially homologous herpesvirus, HSV-2, also resulted in activation of HSV-1. To minimize excessive synthesis of infectious HSV-2 while allowing virus gene expression, repressed HSV l-infected cultures were superinfected with HSV-2 temperature-sensitive (ts) mutants (tsF3, tsB5, or tsH9). The predominant virus replicated after HSV-2 ts mutant superinfection at a nonpermissive temperature was identified as activated parental-like HSV-1 by (i) plaquing efficiency at permissive (34 degrees) and nonpermissive (40.5 degrees) temperatures, (ii) sensitivity to inhibition by the HSV-l-specific antiviral agent (E)-5-(2-bromovinyl)-2'-deoxyuridine, and (iii) restriction endonuclease cleavage analysis. In addition, the fact that superinfection with HSV-2 tsB5 or tsH9, which are unable to synthesize virus DNA and express only early virus genes at nonpermissive temperature, resulted in synthesis of virus demonstrated that HSV-2 DNA synthesis is not required for activation. This system has provided the basis for further studies concerning the regulation of HSV gene expression in human cells. PMID- 6278724 TI - A radioimmunoassay for herpes simplex virus (HSV) thymidine kinase. PMID- 6278725 TI - Interaction of satellite phage P4 with phage 186 helper. PMID- 6278726 TI - Evidence for control of herpes simplex virus mutagenesis by the viral DNA polymerase. PMID- 6278727 TI - Pathway of assembly of herpesvirus capsids: an analysis using DNA+ temperature sensitive mutants of pseudorabies virus. PMID- 6278728 TI - Evolutionary variants of mouse adenovirus containing cellular DNA sequences. PMID- 6278729 TI - Sequence of RNA segment 7 of the influenza B virus genome: partial amino acid homology between the membrane proteins (M1) of influenza A and B viruses and conservation of a second open reading frame. PMID- 6278730 TI - Expression and modulation of virus receptors on lymphoid and myeloid cells: relationship to infectivity. PMID- 6278731 TI - The presence of viral-induced proteins in nuclei from poliovirus-infected HeLa cells. PMID- 6278732 TI - A temperature-sensitive lesion affecting levels of transformation-specific viral RNA in a mutant of avian sarcoma virus PRCII. PMID- 6278734 TI - Biochemical characterization of protein kinase activities associated with transforming gene products of the Snyder-Theilen and Gardner-Arnstein strains of feline sarcoma virus. PMID- 6278733 TI - Genome structure of the defective avian sarcoma virus PRCIV. PMID- 6278735 TI - Effects of 12-O-tetradecanoyl-phorbol-13-acetate on cell proliferation and Epstein-Barr virus DNA replication. PMID- 6278736 TI - Restricted expression of integrated primate type-C virus (gibbon ape leukemia virus-simian sarcoma virus) proviral DNA in nonproductively infected human B lymphoblasts. PMID- 6278737 TI - Endogenous murine leukemia virus-encoded proteins in radiation leukemias of BALB/c mice. PMID- 6278738 TI - Papilloma virus-induced tumors contain a virus-specific transcript. PMID- 6278739 TI - Genomic characterization of a highly oncogenic env gene recombinant between amphotropic retrovirus of wild mouse and endogenous xenotropic virus of NIH swiss mouse. PMID- 6278740 TI - Specific interaction of the SV40 T antigen-cellular p53 protein complex with SV40 DNA. PMID- 6278741 TI - FV-1 restriction of age-dependent paralytic lactic dehydrogenase virus infection. PMID- 6278742 TI - Physical map of the short foldback sequences of herpes simplex virus type 1 DNA. PMID- 6278743 TI - Glycoprotein processing in mutants of HSV-1 that induce cell fusion. PMID- 6278744 TI - Factors determining the susceptibility of NIH swiss mice to erythroleukemia induced by Friend murine leukemia virus. PMID- 6278745 TI - The molecular biology of rotaviruses. II. Identification of the protein-coding assignments of calf rotavirus genome RNA species. PMID- 6278746 TI - Intermolecular association of HANA glycoprotein of Sendai virus in relation to the expression of biological activities. PMID- 6278747 TI - Alterations in the genomes of avian sarcoma viruses. PMID- 6278748 TI - Characterization of bacteriophage phi 42 DNA. PMID- 6278749 TI - Induction of anemia by avian leukosis viruses of five subgroups. PMID- 6278750 TI - Detection of VSV(MuLV) pseudotypes by an immunobiochemical technique. PMID- 6278752 TI - [Clinic and epidemiology of human cytomegalovirus (review) (author's transl)]. PMID- 6278751 TI - Expression of herpesvirus thymidine kinase gene under control of early promoter of SV40. PMID- 6278753 TI - [Tumorgenesis due to oncovirus: special reference to avian sarcoma viruses (review) (author's transl)]. PMID- 6278754 TI - [Isolation of human rotavirus in cell cultures and its antigenic and physicochemical properties (author's transl)]. PMID- 6278755 TI - [Variation of diarrhea viruses detected in Japan with special reference to spherical viruses other than rotaviruses (author's transl)]. PMID- 6278757 TI - Transmitter-related studies in the isolated, perfused eye of the cat. PMID- 6278756 TI - Effect of synaptic transmitter drugs on receptive fields of rabbit retinal ganglion cells. PMID- 6278758 TI - [Use of isofocusing for isolating and characterizing phosphorus metabolism enzymes]. AB - A modified procedure is described for colorimetric estimation of phosphate in a sample containing ampholines; the procedure enabled to simplify the detection of some enzymes of phosphorus metabolism in the fractions obtained after isoelectric focusing of the initial protein mixture. The analyses did nor require an isotope technique and dialysis and were carried out within a relatively short period. PMID- 6278759 TI - [Structural and functional characteristics of the sarcoplasmic reticulum membranes of the skeletal muscles in thyrotoxicosis]. AB - A rate of CA2+ accumulation and efficiency of Ca2+-pump were decreased in the fractions of skeletal muscle sarcoplasmic reticulum isolated from rabbits with thyrotoxicosis. Temperature of phase transition in membranes of sarcoplasmic reticulum was changed by 4-5 degrees towards low temperatures. Under conditions of thyrotoxicosis fatty acid composition of total lipids from the sarcoplasmic reticulum fractions was altered as follows: amount of palmitic acid was increased 1.4-fold but oleic and arachidonic acids were decreased 21.8% and 40.5%, respectively. Modification of Ca2+-transport properties in sarcoplasmic reticulum caused by long-term administration of thyroid hormones into animals appears to alter the membrane protein-lipid composition. PMID- 6278760 TI - [Membrane-bound sialoglycoproteins of the rat liver as affected by the combined action of benz(a)pyrene and sulfur dioxide]. AB - Simultaneous effect of benz(a)pyrene and sulphurous gas was accompanied by distinct alteration in metabolism of sialoglycoproteins in liver tissue; at the time N-acetylneuraminic acid was redistributed in liver subcellular organelles (mitochondria, lysosomes and endoplasmic reticulum) as well as cytochromes P-450 and b5 were induced in liver microsomes. The alterations observed appear to play a definite role in manifestation of the unfavourable effect in combined action of the carcinogen and chemical contaminations of environments. PMID- 6278761 TI - [Spin label analysis of subclasses of high density lipoproteins]. PMID- 6278762 TI - [Effect of experimental hypertension on activity of certain enzymes of energy metabolism in the rabbit aorta]. PMID- 6278763 TI - [Analysis of pleioregulatory disorders in disease]. PMID- 6278764 TI - [Structural-functional analysis of erythrocyte membranes with different content of cholesterol]. PMID- 6278765 TI - [Biochemical mechanisms of regulation of gastric secretion of pepsin]. PMID- 6278766 TI - [Lymphogenic dissemination of ovarian cancer based on autopsy data]. AB - A clinicopathological study of 152 autopsied cases of ovarian cancer was carried out. Metastases in lymph nodes ranked third (57.9%), following lesions in the peritoneum (92.1%) and greater omentum (79.6%). Metastases in lymph nodes were found in 88 cases; almost invariably, they were concomitant with implantation and hematogenous metastases. The sites of lymphogenous lesions were most frequently in paraaortal (72.7%), pelvic (47.7%) and mesenteric (27.3%) lymph nodes. No metastases in lymph collectors were detected in nearly half the cases (42.1%) of generalized malignancy. It was shown that lymphogenous dissemination does not necessarily predominate in the clinical picture even at the terminal stage of the disease. PMID- 6278767 TI - [Potential for benz(alpha)pyrene uptake from the soil into potato tubers and grain]. AB - A significant correlation between benz(a)pyrene concentration in soil and potato tubers was established for its soil level of 0.6-50.0 mg/kg. This points to potato uptake of the carcinogen even at low levels of pollution. Absence of correlation between benz(a)pyrene concentrations in grain (husk) and soil even for relatively high levels of soil pollution (up to 320 micrograms/kg) shows that wheat and millet grains do not absorb the carcinogen. PMID- 6278768 TI - [Role of electron microscopy in determining the degree of differentiation and cell type in oat cell carcinoma of the lung]. AB - Ultrastructurally, small cell carcinoma of lung may be classified as follows: Type I -- undifferentiated small cell carcinoma in which cells do not show any ultrastructural signs of organ- or tissue-specific differentiation. Type II -- differentiated small cell carcinoma consisting of two groups of cells: undifferentiated (without ultrastructural organ-or tissue-specific features) and differentiated (with ultrastructural organ-or tissue-specific features). Type II may occur in different modifications depending on the variety of differentiated cells, e. g. endocrine, mucous, alveolocytic, squamous-cell, mixed, etc. Histologically, small cell carcinoma of lung is represented by a heterogenous group of tumors. Application of electron microscopy provides an objective and reliable means of identification of type and stage of differentiation of cells of this carcinoma. PMID- 6278769 TI - [Role of immunotherapy in the combined treatment of lung cancer]. PMID- 6278771 TI - [Foremost tasks in the problem of trophoblastic disease]. AB - Among the most urgent problems of trophoblastic disease are: (1) timely diagnosis which may be assured by histological examination of material obtained by cutterage from uterine cavity whenever discharge is tinged with blood; (2) obligatory determination of the marker of the disease-chorionic gonadotropin in urine; and (3) chemotherapy with due account of "factors of risk". All these problems may be solved chiefly by organizational means and, first of all, by setting up endocrinological laboratories in city districts. PMID- 6278770 TI - [Treatment of far-advanced stages of lung cancer]. PMID- 6278772 TI - Detection of anti-varicella-zoster virus antibodies in blood donors by automated passive haemagglutination. AB - 1,219 normal blood donors were screened for anti-varicella-zoster virus antibodies by automated passive haemagglutination (APH) on a Technicon 15-channel AutoAnalyzer. 98 samples (8%) were positive by APH and 92% of these were shown to have antibody titres greater than or equal to 1/16 by indirect immunofluorescence assay (IFA). Parallel testing on an additional 265 donors revealed that 11 (4%) individuals had IFA titres greater than 1/16 and 6 of these 11 donors were identified by APH. Thus, APH is an effective means of rapidly detecting units of plasma acceptable for production of varicella-zoster immunoglobulin. PMID- 6278773 TI - [Radioisotope hepatography in assessing the liver function in hypertension and arteriosclerosis]. PMID- 6278774 TI - [Possible formation of Venezuelan equine encephalomyelitis DNA-provirus in tissue culture cells]. PMID- 6278775 TI - [Chronic rubella virus-induced infection of human continuous cells]. AB - A stable chronically rubella virus-infected culture of HEp-2-BK cells existing for over 30 months has been obtained as a result of a single inoculation and further passages. This system is characterized by the lack of cell destruction and permanent production of infectious virus in titres of 2.5-6.3 lg PFU/ml. Synthesis of RNAs of the same classes as in the acute infection was demonstrated in the chronically infected cell culture (CICC) but virion RNA production was less marked. Only a portion of the cells in the population was found to carry the infectious virus. Virus-free cell clones were as susceptible to rubella virus as the control culture. The process of persistence proved to be resistant to virus specific antibody as well as to the effect of higher (40 degree C) or lower (34 degree C) temperatures. Actinomycin D, 5-bromodeoxyuridine, and 5-iodo-2 deoxyuridine exerted no significant effect on virus reproduction in CICC which, combined with negative results of the experiments on transfection of the susceptible cultures with DNA preparations recovered from CICC, evidenced against the role of integration mechanisms in the establishment and maintenance of the HEp-2-BK system. A clear-cut interference with a heterologous virus was demonstrated in HEp-2-BK culture. The formation of the chronic form of infection in HEp-2-BK culture appears to be due to weak cytocidal properties of rubella virus and formation of endogenous interferon. PMID- 6278776 TI - [New antigen induced by herpes simplex virus in human tumors]. AB - The study showed the new antigen induced by herpes simplex virus type 2 (HSV-2) to be present not only in cancer tumors of the cervix and corpus uteri, and ovaries but also in malignant tumors of the mammary glands, kidneys, urinary bladder, as well as in tissues of fibrous-cystic mastopathy and fibroadenoma of the mammary glands. In malignant tumors of the cervix, however, the HSV-2-induced antigen was found more frequently and had a higher serological activity than in other tumors. In contrast to tumors, this antigen was not detected in normal tissues. Among 25 specimens of malignant cervical tumors containing the virus induced antigen, 8 specimens contained virus antigen and 4 yielded biologically active virus. The virus-induced antigen is found in the tumors much more frequently then viral antigen or virus. Detection in the tumors of this new antigen demonstrates the presence in them of viral genetic information coding for the synthesis of this antigen as confirmed by transfection experiments. PMID- 6278778 TI - [Effect of the Tahyna virus on Langat virus persistence in the central nervous system of the mouse]. AB - The features of development of a mixed infection in which one of the infectious agents (Tahyna virus) had a marked immunodepressive activity and the other (Langat virus) caused predominantly asymptomatic course of the disease were studied. Successive infection with these agents increased the severity of the disease (lethality in monoinfection 7-18%, in mixed infection 57%). At the same time the duration of persistence in the brain of mice of Langat virus but not of Tahyna virus increased up to 110 days. It was demonstrated by the method of linear labyrinth that long-term asymptomatic carrier state of Langat virus was accompanied by disorders in the orienting-analysing function of the central nervous system. It is suggested that an increase in the duration of asymptomatic virus carrier state is due to the inhibiting effect of Tahyna virus on the T dependent mechanisms of virus elimination from the body. PMID- 6278777 TI - [Rotavirus gastroenteritis. Laboratory diagnosis and study methods]. PMID- 6278779 TI - [Morphological study of a mixed alphavirus infection]. AB - In combined paired cultivation of 8 Venezuelan equine encephalomyelitis virus, multiploid virions formed in 17.8% of cases. Five clones with this effect were used in mixed infections with clones of Semliki Forest and Sindbis viruses. In these infections changes in virions of the virus progeny were observed in 20% and 16.6%, respectively. Mixed cultivation of pairs of Sindbis and Semliki Forest viruses resulted in formation of multiploid virions in 42.8%. Besides, in two mixed populations formed upon combined multiplication of the latter viruses virions of unusual shapes were found: rounded, oval, elongated, and triangular designated as polymorphic. PMID- 6278780 TI - [Modelling of a chronic infection in a cell culture of the brain of suckling Syrian hamsters due to viruses of the tick-borne encephalitis complex]. AB - The capacity of 3 members of the tick-borne encephalitis virus complex (Langat Tp 21, Sophyin, Elantsev) differing in their biological properties to induce chronic infection in primary cultures of suckling Syrian hamster brain cells (SHB) was studied. Three types of the infectious process were observed in these cells. Langat Tp-21 virus showed cytoproliferative activity in chronically infected SHB cell cultures; formation of cell colonies and alternation of phases of destruction and repopulation were observed with persisting Sophyin strain. The Elantsev strain in these cells produced infection with unestablished virus-cell equilibrium. The persisting viruses were shown to undergo changes in their biological properties consisting in the loss of the hemagglutinating activity and reduced pathogenicity for the susceptible animals. PMID- 6278781 TI - [Damaging action of Venezuelan equine and Eastern equine encephalomyelitis viruses on the chromosome apparatus of mouse bone marrow cells]. AB - Cytogenetical study of bone marrow cells of mice infected with pathogenic and attenuated strains of Venezuelan (VEE) and Eastern (EEE) equine encephalomyelitis viruses was carried out to elucidate the pattern of changes of the chromosomal apparatus in the infected animals, and differences in the effect of strains with different degree of pathogenicity on the cell during mitosis. It was shown that inoculation of mice with pathogenic and attenuated VEE and EEE virus strains led to the appearance in the bone marrow of a larger number of aberrant cells. Both VEE virus strain induced a significant increase both of the total number of aberrant cells and of the cells with true aberrations. The pathogenic and attenuated EEE virus strains also caused a marked increase in the number of aberrant cells, but while the number of true aberrant cells is significant for the pathogenic strain, the attenuated strain causes an insignificant change in this parameter. PMID- 6278783 TI - [Restriction endonuclease study of interstrain differences in the DNA of herpes simplex virus type 1]. AB - Electrophoregrams of the products of cleavage of DNA of 4 HSV-1 strains by restrictases Xbal, Hind III, and Sau 18/4 were compared. The sets of fragments obtained by the effect of Xbal restrictase on DNA of all 4 strains were found to be identical. At the same time, analysis of the products of DNA hydrolysis by Hind III and Sau 18/4 revealed significant interstrain differences. PMID- 6278782 TI - [Malignant transformation of newborn hamster cells by herpes simplex virus type 2]. AB - The oncogenic potentials of HSV-2 inactivated by ultraviolet rays in newborn hamster cell culture were studied. Virus-induced morphological and malignant transformation of cells was accompanied by synthesis of virus-specific antigen, changes in morphology, formation of colonies in semi-liquid agar, and tumorigenicity of cells. The animals bearing tumors induced by transformed cells showed humoral and cellular immune responses to the virus-specific antigen. Lines of transformed and tumor cells were obtained and established in passages. PMID- 6278784 TI - [Practical application of a method for the ultrasonic disintegration of a virus antibody complex in virological research]. PMID- 6278785 TI - The malignant potential of mucinous cysts of the pancreas. PMID- 6278786 TI - [Effectiveness of Zanil and Acedist in the control of fascioliasis in sheep]. PMID- 6278788 TI - Transferrin binding to the membranes of lactating rabbit mammary gland. Iron transfer from membranes to lactoferrin. PMID- 6278787 TI - Screening for lung cancer. AB - The survival from bronchogenic carcinoma is highly dependent upon stage at the time of treatment. This is particularly true for squamous cell carcinoma, adenocarcinoma, and large cell carcinoma, but holds true for small cell carcinoma as well. The problem presented to the medical profession has been to find a practical means of detecting lung cancer while it is still at an early stage. Three studies in progress have indicated that a larger proportion of the patients may be found to have early stage lung cancer when screened with a combination of chest X-rays and sputum cytology. However, the detection of these early stage cases has not yet been translated into an improvement in the overall mortality rate from lung cancer. PMID- 6278789 TI - [Electron microscopic investigations about the influence of the cyclic N6 monobutyryladenosine-3':5'-monophosphate (mb-cAMP) on the neuron differentiation in the explantate cultures of the hippocampus (author's transl)]. AB - 1. The influence of the cyclic N6-monobutyryladenosine-3':5'-monophosphate (mb cAMP) on the neurogenesis was investigated by means of electron microscopic and morphometric methods at the explantate cultures of the fetal hippocampus of the rats in vitro. 2. After three days of the in vitro cultivation the neuroblasts of the treated cultures show not significantly changes of the total number of ribosomes in comparison to the controls. 3. After 7 and 14 days of the in vitro cultivation the neurogenesis is significantly stimulated under the influence of the mb-cAMP. The drug produces a significant increase of the free ribosomes and polysomes in company with a higher degree of differentiation of the granular endoplasmatic reticulum. Under the influence of the mb-cAMP the total number of ribosomes significantly increases by 32% (after 7 days) respectively 20% (after 14 days). 4. The results might suggest, that mb-cAMP might stimulate the protein synthesis in connection with an increased nerve fibre growth in vitro. PMID- 6278790 TI - [New viruses of the family of Papovaviridae isolated from man]. PMID- 6278791 TI - [Myeloperoxidase of neutrophil leukocytes]. PMID- 6278792 TI - [Molecular genetic and cellular basis for the interaction of genetic factors: genes, hormones and receptors]. PMID- 6278793 TI - [Granular-cell Abrikosov's tumor with skin localization]. PMID- 6278794 TI - [Organization of the sensory input of command neurons]. AB - Dynamics of habituation of synaptic responses to tactile stimulation of the snail skin was used as a criterion for defining organization of the receptive field of an escape behaviour command neurone. It was shown that every command neurone had its own specific nonhabituating area of the receptive field while the size of the field was common to all command neurones - it comprised the whole skin surface. The receptive field of a command neurone system is not homogeneous in structure. Areas corresponding to ecologically significant regions can be singled out by the maximal response amplitude and low rate of habituation. PMID- 6278795 TI - [Melanogenesis and postnatal cellular proliferation in the retinal pigment epithelium in the rat]. PMID- 6278796 TI - Three rare ovarian tumours: Krukenberg, struma ovarii and granulosa cell neoplasms. PMID- 6278798 TI - Caprine herpesvirus infection in Switzerland: some aspects of its pathogenicity. PMID- 6278799 TI - [Virus-like structures in human gastric cancer]. PMID- 6278797 TI - [Calcium lithiasis II. Idiopathic or hypophosphatemic hypercalciuria? Vitamin D - metabolism and othophosphate therapy]. AB - Hypophosphateamia in patients with Ca lithiasis leads to the activation of the vitamin D endocrine system: the plasma 1.25(OH)2-D3 concentration is raised. The raised 1,25(OH)2-D3 biosynthesis causes an increase in intestinal Ca absorption, which in its turn explains the hypercalciuria. The syndrome originally presented by Albright and his pupils as "idiopathic hypercalciuria" in fact corresponds to a secondary, reactive D hypervitaminosis. According to the present findings the often wrongly used term "so-called idiopathic calciuria" should be replaced by the pathogenetically correcter term "hypophosphataemic calciuria". Efficient treatment of this syndrome consists in sufficient oral orthophosphate substitution. In the interests of a better understanding as a requirement for suitable treatment of this metabolic disorder, the vitamin D metabolites and their renal key enzymes, 25-OH-Vitamin-D3-1-Hydroxylase and 25-OH-Vitamin -D3-24 Hydroxylase, are described. The hormonal control add the activation and inactivation of the vitamin D endocrine system are explained independently of the individual Ca and phosphate requirement of the organism. The dependence of renal Ca excretion on the rate of glomerular filtration is pointed out once again. The clinical-diagnostic term hypercalciuria must not be applied globally but individually. Indications, counter-indications, dosage, duration and side-effects of the orthophosphate therapy are discussed. PMID- 6278800 TI - Cytodiagnosis of herpes simplex virus infection in the newborn infant: report of a case. AB - Typical herpetic stomatitis that developed in a premature 6-day-old infant was initially diagnosed cytologically. The cytomorphologic characteristics of herpes simplex virus (HSV) infection in smears of scrapings from the oral mucosa helped to establish the diagnosis of a neonatal HSV infection, permitting early treatment with cytosine arabinoside (ara-C) and subsequent complete recovery from the generalized infection. The diagnosis was later confirmed by rises in the serologic titer of complement-fixing antibodies for HSV type 2. PMID- 6278801 TI - Cytologic diagnosis of occult small-cell undifferentiated carcinoma of the lung. AB - Small-cell undifferentiated carcinoma of the lung is usually metastatic and/or locally advanced at diagnosis. We describe an occult tumor that cytologic techniques diagnosed in a minimally symptomatic, high-risk individual. Despite the absence of radiologic or histopathologic confirmation, bronchoscopically directed cytologic specimens localized the lesion. A tiny, apparently stage I, small-cell undifferentiated carcinoma was successfully resected. PMID- 6278802 TI - Serum angiotensin-converting enzyme and blood monocytes in splenectomized individuals. AB - In 45 splenectomized individuals without malignancies, residual splenic tissue was detected by 99mTc-scanning using reinjection of labelled heat-damaged and autologous erythrocytes. The angiotensin-converting enzyme in serum (SACE) and the blood monocyte count were measured in order to demonstrate an eventual association between these parameters as well as the influence of residual splenic tissue. In 20 patients with detectable spleen tissue SACE was normal (26.2 +/- 4.4 U/ml), but in 25 patients without residual spleen tissue SACE was slightly elevated (28.7 +/- 6.4 U/ml; p less than 0.02) as compared with healthy controls (24.4 +/- 6.2 U/ml). In both groups of patients blood monocytes were markedly increased. However, no correlation could be demonstrated between SACE and the monocyte count in any group. In 2 patients SACE was above the upper limit of the normal range (greater than 36.8 U/ml) as found in sarcoidosis but no signs of this disease could be demonstrated. It is concluded that splenectomy must be taken into account when the significance of raised SACE is evaluated. It may be suggested that the spleen exerts an effect on the metabolism of SACE or on the ACE-producing cells in the organism. PMID- 6278803 TI - Opiate action on adenohypophyseal hormone secretion during anesthesia and gynecologic surgery in different phases of the menstrual cycle. PMID- 6278804 TI - Elements of structure and ultrastructure of the blood-thymus barrier in ACTH involuted thymus. AB - The ACTH influence upon the thymus may be a reliable model for stress involution. In this case, the cortical lymphocytic depletion is accompanied by mast cell accumulation and increased caliber of the blood vessels. The blood-thymus barrier which has an active role in involution shows an enriched transport activity of the endothelial cells, great enlargement of the basement membrane, increased macrophage activity within the perivascular space with elevated values of acid phosphatase activity, and thickening of the fibrillar network. The epithelioreticular cells show plenty of vacuoles in their cytoplasm, the mitochondria undergo swelling processes, and their cristae are diminished. The ultrastructural data show that lymphocyte depletion is carried out by macrophage mediated lymphocytolysis. But by counting the peripheral blood cells an earlier mechanism is revealed; i.e. migration through the enlarged but more permeable blood-thymus barrier. The epithelioreticular cells do not seem to have an active, direct implication in any of the phenomena. PMID- 6278805 TI - Conformation-activity relationships of corticotropin segments. PMID- 6278806 TI - Steroid spectrum in human urine as revealed by gas chromatography. IV. Changes in the exception of 16-oxygenated neutral steroids by children with 21-hydroxylase deficiency during different stages of development. PMID- 6278807 TI - The Ca2+-sensitive K+ transport in inside-out red cell membrane vesicles. AB - Ca2+-sensitive K+ transport ("Gardos-effect") was investigated in inside-out red cell membrane vesicles (IOVs) by measuring the effect of Ca2+ and other agents on the 86Rb+ (K+-analogue) influx. 50 micro M-5 mM Ca2+ stimulated the 86Rb+ influx as compared to the effect of 1 mM EDTA. Mg2+ and chlorpromazine inhibited the effect of Ca2+, oligomycin and quinine were without effect. The permeability increase of K+, elicited by Ca2+ in IOVs, corresponded to that of intact cells. The effect of Ca2+ was not specific to K+ with respect to Na+. The Ca2+-sensitive K+ channels saturated about 5-10 mM KCl in the medium. The results indicate that IOVs the Ca2+-sensitive K+ transport has altered characteristics as compared to that of intact cells and resealed ghosts. However, in contrast to the suggestion of other investigators, it was found that some features of this phenomenon are still preserved in IOVs. PMID- 6278808 TI - Correlation between acetylcholine-evoked electrical activity, effect of cyclic AMP and actual redox state in frog rectus muscle. AB - In view of the very high correlation coefficient between actual redox state potential level in the biophase and spike activity--triggered by acetylcholine and influenced by cAMP--in skeletal muscles it seems rather reasonable the conclusion that the redox state potential level influences the cooperative effects not only in the case of acetylcholine but, directly, also the actions of cAMP, since the type of the regulatory influence of actual redox state potential level is non-additive in some cases, though additive in others. PMID- 6278809 TI - Drinking habits and peripheral alcoholic neuropathy. AB - Drinking habits of 156 consecutive polyneuropathic and 106 consecutive pressure palsy patients were evaluated in retrospect. Respectively, 46 patients (30%) had alcohol polyneuropathy and 32 (30%) got pressure neuropathy while being drunk and these patients were analyzed in more detail. Most of the patients with alcoholic neuropathies were men, those with polyneuropathy being older than those having pressure palsies. Pressure neuropathy coincided with alcoholic polyneuropathy in 13 patients (28%). Other medical complication of heavy alcohol drinking (i.e. liver diseases, seizures and cerebellar signs) were seen in 54% of the patients with polyneuropathy and in 6% of the patients with pressure palsies. Heavy drinking prolonged the disability due to pressure palsy. The present study confirms the significant role of alcohol abuse in etiology of peripheral neuropathies. Heavy drinking seems to worsen the prognosis of these neuropathies. PMID- 6278810 TI - Lectin-binding sites of the human pituitary adenoma cells by means of the ferritin-labeling technique. PMID- 6278811 TI - Status marmoratus and Bielschowsky bodies. A report of two cases and review of the literature. AB - Intraneuronal inclusions, consisting of polyglucosan and having histochemical and ultrastructural features identical to Lafora body of familial myoclonic epilepsy (Unverricht-Lafora disease), have been found restricted to the lateral pallidum in five patients. Two of these patients were also found to have status marmoratus of the basal ganglia. These lateral pallidal inclusions have been named after Bielschowsky, their original discoverer. We report two additional patients with status marmoratus and Bielschowsky bodies and suggest that these two conditions are frequent concomitant phenomena arising independently from a common cause. PMID- 6278812 TI - Light- and electron-microscopic studies of intracytoplasmic acidophilic granules in the human locus ceruleus and substantia nigra. AB - Using the brains of 30 patients with mental and neurologic disorders, we studied the intracytoplasmic acidophilic granules in neurons of the substantia nigra and locus ceruleus by light and electron microscopy. The granules were present in all 30 brains, including those with no recognizable pathologic change, there was no correlation between their appearance and the age, sex, disease of, or the medication received by, the patients. In four electron-microscopically examined brains, we noted many small, round electron-dense bodies in the perikarya and neuronal processes of the substantia nigra and locus ceruleus. The bodies were packed tightly within a double membrane; in shape, size, and distribution in the neuronal cytoplasm, they corresponded to acidophilic granules. Some mitochondrial matrices contained one or more similar, but smaller inclusion bodies; large bodies pushed aside the mitochondrial cristae. We conclude that the acidophilic granules represent high developed forms of mitochondrial inclusions. PMID- 6278814 TI - Papovavirus-related RNA sequences in human neurogenic tumours. AB - Thirty-two human brain tumours were investigated for the presence of papovavirus related RNA sequences. Radiolabelled viral DNAs isolated from SV40, BKV, HDV, and BPV were hybridized in-situ on to tumour kryostat sections under conditions that detect complementary RNA. SV40-related RNA was detectable in 34% of all tumours tested but was not found in normal human brain tissue. SV40-positive tumours included 1/1 angiofibroma, 1/4 astrocytomas, 2/4 metastatic brain tumours, 1/1 melanoma, 3/7 meningiomas, 2/4 neurinomas, 1/2 oligodendrogliomas. Bovine papilloma virus-, BKV-, and HDV-related RNAs were absent from all tissues tested. The presence of SV40-related nucleic acids in a large number of human neurogenic tumours may reflect a viral involvement in the etiology of these tumours. PMID- 6278813 TI - Adenovirus-related RNA sequences in human neurogenic tumours. AB - Thirty two human tumours, mainly neurogenic, have been investigated for the presence of adenovirus-related RNA sequences. 3H-labelled tumour virus DNA probes derived from human adenoviruses types 2 and 12, bovine adenovirus type 3, and avian adenovirus CELO were hybridized in-situ on tumour kryostat sections under conditions that detect complementary RNA. Tumour virus-related RNA was detected in 62% of all tumours tested, but was not detectable in normal human brain tissues. Expression of tumour virus-related RNA was found in 2/4 astrocytomas, 2/4 metastatic brain carcinomas, 2/2 glioblastomas, 1/1 melanoma, 5/7 meningiomas, 4/4 neurinomas, 1/2 oligodendrogliomas, and 1/1 rhabdomyosarcoma. The presence of adenovirus-related RNA in the majority of human neurogenic tumours may reflect a viral involvement in the pathogenesis of these tumours. PMID- 6278816 TI - 99mTc-pertechnetate uptake after total knee replacement in rheumatoid arthritis. AB - In 6 out of 48 non-hinged knee joint replacements in 35 patients, 99mTc04- uptake values were clearly increased compared with a control group. Measurements were performed an average of 3 years (range 9 months - 4 1/2 years) after operation. Four of the six cases showed loosening of the prosthesis at arthrotomy and one showed an infection. Tests for hypersensitivity to the different components of the prosthesis and cement were normal. Histology and immunofluorescence of the synovial membrane obtained at arthrotomy in the four operated patients showed no recurrence of the synovitis. This study indicates that there is no recurrence or only a slight recurrence of rheumatoid synovitis after total knee joint replacement of the geometric type. PMID- 6278815 TI - Anderson-Fabry's disease: neuropathological and neurochemical investigation. AB - A clinical, neuropathological and neurochemical study of a case of Anderson Fabry's disease is described. The clinical course mainly consisted of repeated ictus with major involvement of the CNS. The neuropathological examination is dominated by severe alterations in the cerebral vessels due to glycolipid deposits on the walls, with reduction or occlusion of the lumen. This is correlated with secondary ischaemic foci scattered throughout the cortex as well as through the white matter. In addition, the cells of the cerebral cortex, thalamus, basal ganglia, amygdala, cerebellar and olivary nuclei show a marked accumulation of lipofuscin. Biochemical examination reveals a threefold increase in galactolipids due to the specific alpha-galactosidase deficiency. Cholesterol is reduced secondarily to ischaemic myelin damage. Glycosaminoglycans uronic acid is increased in cytosol and membrane-bound fractions which could be related to reactive gliosis. Glycoprotein sugars show a decrease in N-acetyl-neuraminic acid and fucose as well as an increase in hexosamines and hexoses in membrane-bound fraction, while in cytosol fraction all sugars are increased. This suggests that the alpha-galactosidase deficiency can alter not only the glycolipid but also the glycoprotein metabolism, resulting in a higher presence of hexosamines and hexoses-rich glycoproteins. PMID- 6278817 TI - Long-term social prognosis after hip fractures. AB - A long term follow-up study of 518 patients with hip fractures was undertaken 2.5 years after the operation. The total mortality was 35 per cent (180/518). The risk of social deterioration for patients admitted from home was 47 per cent (132/281). A statistical analysis showed this risk to be determined primarily by the age of the patient and secondarily by the placement of the patient on discharge from hospital. A thorough description of these factors is presented. PMID- 6278818 TI - The action of putative neurotransmitter substances in the cat labyrinth. AB - Possible neurotransmitter candidates were tested in the labyrinth of the cat with the aid of microiontophoretic techniques. Depending on the recording site, spontaneous regular or irregular fibre activity was obtained in the subsynaptic region of the macula sacculi. Ejection of GABA enhances the firing rate, whereas acetylcholine reduces the spontaneous activity. A similar application of glycine and proline produced no effect. The action of GABA was specifically blocked by the GABA antagonists bicuculline and picrotoxin. The alkaloids further induced a decrease in the spontaneous activity which lasted for several minutes. PMID- 6278819 TI - Fine structure of-and some histochemical observations on-chondroid regions of benign mixed parotid tumours. AB - Seven cases of mixed parotid tumour were studied at the light-and of these, three at the electron microscopical level. In the latter group, special attention was paid to the fine structure of the chondroid regions of the tumours. The chondroid regions showed a typical cartilage fine structure-of greater interest was the finding that this tissue was mixed type of both hyaline and elastic cartilage. This was evident with respect to both cell and extracellular matrix. Two cell types were found: (A) cells similar to cells of hyaline cartilage and (B) cells resembling cells shown to be involved in synthesis of elastic fibres (elastic tendon fibroblasts, elastic cartilage chondrocytes and arterial smooth muscle cells). As in normal calcifying cartilage the chondroid regions contain numerous membrane-delimited matrix vesicles. However, mineral deposits within the latter were rare, indicating either a suppressed tendency towards mineralization of a lower stage of tissue maturation at which the mineralization process has not yet started. PMID- 6278820 TI - The ultrastructure of the geniculate ganglion. AB - The ultrastructure of the geniculate ganglion was studied in the guinea pig and monkey. Two cytologically distinct colonies of ganglion cells were seen and identified as light and dark cells. All neurons were unmyelinated and most were unipolar, although a few bipolar cells were seen. The axons, upon leaving the cell bodies, followed convoluted courses in the vicinity of the cells bodies before becoming invested by myelin sheaths. Two types of nerve terminal were identified on the small light cells. The first type contained predominantly agranular and spherical vesicles and had synaptic contact with the cell body. The second type did not show synaptic contacts but contained many ellipsoidal vesicles. This study provided no information on the functional significance of these nerve terminals. PMID- 6278821 TI - Presacral vipoma in a 16-month-old child. PMID- 6278822 TI - Systemic amyloidosis and non-hematologic malignancy in a large autopsy series. AB - A series of 24,388 consecutive autopsies yielded 148 cases of amyloidosis, for which the associated diseases were tabulated. 13 solid primary malignancies were found in eleven patients, including one mucoepidermoid cancer of the parotid and one thymoma; two out of eleven had evidence of pyelonephritis. Five patients suffered from past or present renal adenocarcinoma, the most common carcinomatous cause of amyloidosis. Systemic amyloid deposits thus occurred in 2.1% of autopsied patients with renal carcinoma, but showed no obvious correlation with tumor stage or histologic type. Details are presented of an unusual case of hypernephroma producing a stable bronchial metastasis cuffed by nodular amyloid and dissociated by a lymphoid infiltrate containing plasma cells. The occurrence of systemic amyloidosis in non-hematologic malignancy thus appears to be a rare event, which, especially in the case of renal cancer, is assumed to be due to amyloid-fibril protein AA. PMID- 6278823 TI - An EBV-associated, Swedish case of Burkitt-type malignant lymphoma. AB - Morphological, serological, DNA-homology and immunofluorescence studies of a rapidly fatal malignant lymphoma in a 25-year-old Sweden male revealed most of the characteristics of an EBV-associated Burkitt lymphoma. The various findings are discussed in relation to previously reported observations on endemic (African) and non-endemic Burkitt lymphomas. PMID- 6278824 TI - The use of Staphylococcus aureus rich in protein A in the detection of herpes simplex virus antigens. AB - Herpes simplex virus (HSV) type 1 antigens were detected in infected human embryonic lung cells with the aid of specific antiserum and Staphylococcus aureus rich in protein A. When such staphylococci carrying specific anti-HSV IgG on their surface were interacted with various suspension of virus, a reduction in the initial virus titre of about 65% was obtained. However, no direct coagglutination was observed between cell-free supernatants of HSV or HSV infected cells and sensitized staphylococci. When monolayers or suspended cells infected with the virus were treated with dilutions of specific anti-HSV antiserum followed by non-sensitized staphylococci (indirect method), an "aureola" of the bacteria was detected around the cells expressing the viral antigens. A similar picture was observed when infected cells were interacted directly with sensitized staphylococci. Viral antigens were detected already 12 hours post infection, well before the appearance of cytopathic effect. The sensitivity of the indirect method was found to be higher than that of the direct one and dependent on the multiplicity of infection and the serum dilution used. The method is proposed as a rapid means of identifying viral antigens in diagnostic and experimental virology. PMID- 6278825 TI - C1q binding and complement activation by capsular and cell wall components of S. pneumoniae type XIX. AB - Cell wall components (purified cell walls, teichoic acid and residual cell walls) from S. pneumoniae type XIX showed antibody independent C1q binding capacity, as assessed by C1q deviation test, with teichoic acid being the most efficient. Specific capsular substance did not bind C1q. All substances tested produced C1 activation in normal human serum, but not in hypo-gamma-globulinemic serum. Thus, teichoic acid showed high C1q binding capacity but did not activate C1 in the absence of antibodies. Teichoic acid was an effective activator of alternative pathway. Specific capsular substance did not activate the alternative pathway in C1q deficient serum or in Mg2+ -EGTA chelated normal serum. PMID- 6278826 TI - Quantitation of proenzyme C1r-C1s complexes in serum by electroimmunoassay. PMID- 6278827 TI - [Comparative study of the effects of dimethylcurine methylchloride and D tubocurarine on isolated ganglion and phrenic nerve-diaphragm preparation (author's transl)]. PMID- 6278828 TI - [Mode of action of dimethyl l-curine dimethochloride on neuromuscular transmission (author's transl)]. PMID- 6278829 TI - [Radioimmunoassay for guanosine 3', 5'-cyclic phosphate (cGMP) (author's transl)]. PMID- 6278830 TI - [Effect of dl-demethylcoclaurine on cultured rat heart cells (author's transl)]. PMID- 6278831 TI - [Effect of dl-demethylcoclaurine on plasma cAMP in mice (author's transl)]. PMID- 6278833 TI - Studies on the mechanism of the GABAergic inhibition of TSH secretion in male rats. AB - The mechanism of the GABAergic inhibition of the TSH cold-response was studied in male rats. Picrotoxin, a putative GABA receptor antagonist (4 mg/kg but not 0.5 or 2 mg/kg intraperitoneally) significantly decreased the cold-stimulated TSH secretion but did not modify the TRH-induced (100 ng intraperitoneally) TSH response. The inhibitory effects of two GABAergic drugs muscimol (1 mg/kg subcutaneously) and aminooxyacetic acid (10 mg/kg intraperitoneally) were not antagonized by picrotoxin (1-4 mg/kg intraperitoneally), by phenoxybenzamine (2.5 or 5 mg/kg intraperitoneally), by two 5-HT antagonists cyproheptadine (5 mg/kg intraperitoneally and R 41468 (2.5 mg/kg intraperitoneally) or by pimozide (0.5, 1.5 or 2.5 mg/kg subcutaneously). The results suggest that neither picrotoxin sensitive GABA receptors nor noradrenaline, 5-HT or dopamine pathways seem to be involved in the inhibitory action of GABAergic drugs on the TSH cold-response in male rats. PMID- 6278832 TI - On the role of cyclic nucleotides in the regulation of cardiac contractility and glycolysis during hypoxia. AB - A possible involvement of cyclic nucleotides (cAMP and cGMP) in the regulation of cardiac contractility and glycolysis during hypoxia was examined in spontaneously beating rat atria. A reduction of the high oxygen saturation (HiOxSa) of the incubation medium from 95-100% to half produced a rapid decline of the amplitude. The deterioration of 50% was seen after 30 sec. of hypoxia. The decline was partly antagonized by noradrenaline (NA, 1 X 10(-6) M) or hypercalcaemia (5.7 X 10(-3) M instead of 1.9 X 10(-3) M). The cAMP level remained unchanged during the first 12 min. of hypoxia, but the cGMP content increased gradually and reached a significantly increased level in 4-8 min. Paradoxically, the production of lactate decreased, after 30 sec. of hypoxia, but accelerated then 2-4 min. after the onset of hypoxia. The depletion of creatine phosphate and ATP stores was initiated after 2 min. of hypoxia. The arterial content of the active forms of phosphofructokinase and lactate dehydrogenase gradually rose during hypoxia. Sodium nitroprusside (SNP, 1 X 10(-4) M) and NA produced increases in cGMP and cAMP levels, respectively, both in HiOxSa and hypoxia. SNP induced a slight and NA a marked increase in the amplitude in HiOxSa. Verapamil (1 X 10(-6) M) decreased the contractility, but did not affect the levels of cAMP and cGMP. Both SNP and verapamil decreased the lactate production, but they could not resist the NA-induced increase in the atrial lactate level. Hypercalcaemia increased the amplitude but slightly reduced the lactate production in HiOxSa. 45Ca-uptake was reduced to about 35 per cent of control as measured between 5 and 10 min. of hypoxia. It is concluded that the lack of oxygen could have direct and parallel effects on the sarcolemma and on the mitochondria. The former could result in the deterioration of contractility and the latter in the termination of aerobic energy production. Cyclic nucleotides are not involved in either of these phenomena. However, at the low rate of anaerobic glycolysis, e.g. in HiOxSa or at the very early stage of hypoxia, cGMP could inhibit and cAMP accelerate the lactate production. PMID- 6278834 TI - Investigations on the occurrence of cytidine 3',5' monophosphate (cCMP) in tissues. AB - A specific and sensitive radioimmunoassay (RIA) for the detection of cCMP was developed. Antibodies were prepared by immunization of rabbits with succinyl cCMP conjugated to human albumin. In the RIA commercially available 3H-cCMP was used as labeled antigen. Low amounts (0.4-2.6 pmol/g w.wt.) of immunoreactive material could be detected in extracts of brain, heart, ileum, kidney, liver, lung, spleen, testis and uterus from rats, guinea-pig and frogs. The immunoassayable material in rat liver was subjected to column chromatography on AG 1-X8 anion exchange resin. The radioimmunoassayable material isolated from the liver did not cochromatograph with authentic tritiated or unlabeled cCMP. The chromatographic properties of the immunoreactive material did not change in regenerating livers - following partial hepatectomy, or after pronase treatment. However, after treatment with pronase the amount of assayable material, extracted from the rat liver, increased. It is suggested that cCMP - as such - is not present, in detectable amounts, in extracts prepared by commonly used procedures for isolation of cyclic nucleotides. The chemical nature of the cCMP immunoreactive material isolated from rat liver is at present unclear. PMID- 6278836 TI - Studies on hydragogue drugs: effect of surfactants on cAMP levels in the rat jejunal mucosa in short time experiments in vivo. AB - Theophylline and the surface active agents specified below were instilled into the jejunum of anaesthetized rats, and the cAMP levels in the mucosal tissue determined after 71/2 and 15 min. incubation in vivo. Most experiments were done in rats prepared with two tied intestinal loops; one of these served as the control loop and the other as the stimulated loop. The surfactants (mmol/l) included dodecylsulphate (17), dioctylsulphosuccinate (5.6), cetrimonium bromide (4.1), deoxycholic acid (2.4 and 3.6) and Lubrol WX (0.5% w/v). Theophylline (10 mmol/l) caused a substantial increase in cAMP (110% +/- 17, n = 7 and 60% +/- 8.9, n = 10, respectively) and dodecylsulphate a minor and transitory increase (28.1% +/- 3.8, n = 10 and 11.7% +/- 4.9, n = 8). The other agents were without stimulatory effect on cAMP, although like dodecylsulphate they may significantly affect normal intestinal cation and glucose transport under these conditions. The results, therefore, do not suggest that stimulation of cAMP and intestinal secretion induced thereby is any significant phenomenon in the overall hydragogue effect of these agents, at least not in short term jejunal experiments. PMID- 6278835 TI - Net sodium and glucose transport in the jejunum, ileum and colon of anaesthetized rats in response to intraluminal theophylline and anionic surfactants. AB - Osmotically balanced solutions of sodium chloride and glucose (5-150 mmol/l) were instilled into rats prepared with two tied intestinal loops (jej-col or il-col). Luminal accumulation or disappearance of Na and glucose after 15 min. was determined, and the parameters of the linear regression lines of net Na flux (y) with initial Na concentration (x) calculated. Control cation and glucose transport were changed by dioctylsulphosuccinate and dodecylsulphate in the way described by Sund & Matheson (1978). Theophylline (10-25 mmol/l) on the other hand did not alter glucose disappearance, and had a distinctly dissimilar effect on net Na transport compared to the surfactants. This could be described as a parallel displacement of the control regression lines to the right, without improvement in correlation. This effect of theophylline was greatest in the ileum, where mean luminal Na concentration corresponding to zero net transport was raised from about 70 mmol/l under control condition to values above 200 mmol/l. The results are consistent with the view that theophylline mainly affects transcellular and the surfactants mainly paracellular sodium transport, and do not support the theory that the effect of dioctylsulphosuccinate on intestinal transport is related to an activation of the mucosal adenylate cyclase/cAMP system, at least not in short term experiments. PMID- 6278837 TI - Investigations on possible presynaptic effect of adenosine and noradrenaline on cholinergic neurotransmission in guinea pig trachea. PMID- 6278838 TI - Tricaine (MS-222) induced modification on the metabolism of foreign compounds in the liver and duodenal mucosa of the splake (Salvelinus fontinalis X Salvelinus namaycush). AB - The splake, a popular game fish, is a crossbreed which must be reared in nurseries. The fish are marked under anesthesia for later study. We analyze the effect of a common anesthetic, tricaine (MS-222), on the metabolism of foreign compounds in the liver and duodenum of the splake. In the liver and to some extent in the duodenum, aryl hydrocarbon hydroxylase and epoxide hydrase activities were reduced during treatments. The ethoxycoumarin O-deethylase activities were not affected in either the liver or duodenum. Tricaine significantly decreased the hepatic UDP-glucuronosyltransferase activity. The decrease was greater when the aglycone used was p-nitrophenol than with methylumbelliferone. A similar effect was also found after trypsin treatment of the microsomes. No significant decrease in the UDP-glucuronosyltransferase activity was detected in the duodenal mucosa. This was the case when both p nitrophenol and methylumbelliferone were used as aglycones. PMID- 6278839 TI - Differences between alpha-adrenergic and beta-adrenergic inotropic effects in rat heart papillary muscles. AB - alpha-And beta-adrenergic inotropic effects have been shown to be qualitatively different. In order to further characterize these difference we compared the mechanical response to alpha- and beta-adrenoceptor stimulation, respectively, in electrically driven left ventricular papillary muscles from rat heart. The muscles were stimulated by either isoprenaline (Beta-adrenoceptor stimulation), phenylephrine in the presence of propranolol (alpha-adrenoceptor stimulation) or phenylephrine alone (combined alpha-and Beta-adrenoceptor stimulation). Isometric tension (T), rate of rise and decline of tension (first derivate=T') and rate of transition from tension rise to tension decline (negative part of second derivative=T') were recorded. These recordings disclosed qualitative differences between the alpha-and Beta-inotropic response both in dose-response and time course experiments. Maximal Beta-adrenoceptor stimulation caused a small increase in Tmax (18%), intermediate increases in T'max (45%) and T'min (68%) and considerable increase in T'min (145%) ("Beta-type" effect). Maximal alpha adrenoceptor stimulation increased all qualities by about the same degree (23-24% ("a-type" effect). While Beta-adrenoceptor stimulation gave a dose-dependent and pronounced increase in the ratio T"min/T'max (relaxation-onset index), alpha adrenoceptor stimulation decreased it to subcontrol values and phenylephrine alone gave a small dose-dependent increase at higher dose. The time course of the alpha-adrenoceptor stimulation was characterized by a transient decrease in all qualities followed by an increase which reached maximum at 4-5 min. Beta Adrenoceptor stimulation gave a monophasic response which reached maximum after 1 2 min. Phenylephrine alone gave mainly an "a-type" effect although T"min increased significantly more in the absence than in the presence of propranolol and T"min/T'max showed a small increase which developed slowly. Thus Beta adrenoceptor stimulation activated relaxation compared to contraction by a higher degree than did alpha-adrenoceptor stimulation. This probably reflects different mechanisms of action. While the alpha-effect may rely primarily on an increased calcium influx, the Beta-effect probably is the final result of several subcellular effects of cyclic AMP. PMID- 6278840 TI - Leucocytic and haemostatic reactions of the Indian catfish, Heteropneustes fossilis, subjected to environmental pollution by sewage, fertilisers and insecticides. PMID- 6278841 TI - Participation of the adrenergic receptors in the regulation of large bowel motor activity in sheep. AB - In sheep with long-standing fistulae of the caecum, proximal loop of the colon and rumen the motor activity of these parts of the intestine was studied using the balloon method. Slow isoprenaline infusion caused first inhibition of caecal contractions, followed by gradually decreasing intensity of contractions of the colon and rumen. This inhibitory effect of isoprenaline was eliminated by propranolol. Intravenous infusion of phenylephrine failed to inhibit the colonic motor activity. A small dose of atropine, similarly as isoprenaline, caused an irregular inhibition of the contractions of the caecum, colon and rumen. In sheep, in contrast to other animal species, the effects inhibiting large intestine motility are mediated only by the beta-adrenergic receptors. The inhibitory effect of the beta-adrenergic receptors was strongest in the caecum. PMID- 6278842 TI - Decrease of oxygen difference between arterial blood and cerebrospinal fluid after intravenous injection of sodium bicarbonate in hyperoxic patients, anaesthetized, paralyzed and artificially ventilated. AB - In the subjects being prepared to neurosurgical treatment an i.v. injection of NaHCO3 (2 mEq/kg) elicited a significant increase in PCSFO2 from 69 +/- 6.4 (SEM) Torr to 75.5 +/- 3.9 (SEM) Torr. This change ws accompanied by a significant drop of PaO2 from 150.5 +/- 6.0 Torr to 138.0 +/- 5.8 Torr. Metabolic alkalosis (pH 7.54 +/- 0.02 SEM) elicited by bicarbonate administration was accompanied by arterial blood hyperoxia. Both these factors reduce the cerebral flow (CBF). We suppose that changes in the blood--CSF oxygen relationship reflect the presence of a mechanism which might protect the CNS against a decrease in CBF. PMID- 6278843 TI - Biosynthesis and distribution of ACTH and related peptides. PMID- 6278844 TI - Organization of the nervous structures involved in the control of the anterior pituitary. PMID- 6278845 TI - [Role of histamine in the body reactions during excessive central nervous system loading]. PMID- 6278846 TI - Invasive squamous cell carcinoma of the uterine cervix. IV. Analysis of a histopathologic malignancy grading system and construction of a partial index. AB - A new histopathologic malignancy index has previously proved capable of permitting discrimination of patients with carcinoma of the uterine cervix (stage II) with respect to prognosis. Eight histopathologic items graded from 1 to 3 add up to constitute the malignancy index. By regression analysis the number of items could be reduced to 4. Seemingly without loss in the prediction value a partial index was elaborated by the following 4 items: mitosis, mode of invasion, cellular response, and vascular invasion, the latter attributed double the weight. The predictive value of the partial index seemed to be as good as that of the previous malignancy index on an independent materials as well. PMID- 6278847 TI - Results of radiation treatment in carcinoma of the oral cavity. AB - Treatment results in 391 patients with oral carcinoma after radiation therapy are reported. In the total series 5- and 10-year actuarial survival were 30.8 and 25.2 per cent, respectively. After irradiation alone the corresponding figures were 21.5 and 18.5 per cent, after combined radiologic and surgical treatment 33.3 and 28.7 per cent, and after interstitial radium therapy 56.1 and 43.8 per cent, respectively. These differences were mainly an expression of selection factors. Tumor doses above 60 Gy gave a higher survival than lower doses. The presence of neck node metastases was prognostically unfavourable. PMID- 6278848 TI - Adjuvant radiation therapy compared with cyclic chemotherapy in patients with mammary carcinoma. II. Changes of mitogen responses of blood lymphocytes. AB - Blood lymphocyte reactivity to purified protein derivative of tuberculin (PPD) and phytohaemagglutinin (PHA) was examined in 62 patients with breast carcinoma who received postoperative adjuvant radiation therapy or cyclic chemotherapy with chlorambucil, methotrexate and 5-fluorouracil. Both treatments impaired the immunologic reactivity of blood lymphocytes as measured by PPD and PHA stimulation in vitro. Radiation therapy seemed to cause more profound and protracted suppression of the PPD response than chemotherapy. PMID- 6278849 TI - Bilateral carcinoma of the breast. Epidemiology and histopathology. AB - A history of previous carcinoma in the contralateral breast was found in 66 of 1351 women (5%) consecutively diagnosed as having breast carcinoma. The mean time interval between the diagnosis of the first and second tumour was 10.0 years (range 0-37 years); 63 of the tumours were metachronous. No significant differences were found between the first and the second carcinoma with respect to the Ackerman malignancy grading or the frequency of axillary node involvement. High risk groups for bilateral disease could not be defined on the basis of information about familial occurrence, parity, age at first birth, or malignancy grade. PMID- 6278850 TI - Cervical lymph node metastases from an unknown primary tumour. AB - Forty-nine cases of cervical lymph node metastases from an unknown primary tumour have been reviewed. All the patients were treated by radiation therapy. In 7 cases the primary tumour was detected from 4 to 53 months after the end of treatment. The crude survival rates at 2 and 5 years were 39 and 32 per cent, respectively. The survival rate was correlated to sex, microscopic appearance, stage, and extent of the nodal involvement. PMID- 6278851 TI - Effect of glutaurine and its derivatives and their combinations with radiation protective substances upon irradiated mice. AB - The radiation protective effects of glutaurine (gamma-L-glutamyl-taurine, Litoralon), and of some of its derivatives, as well as of their combinations with substances of the amino-alkyl-thiol group, have been investigated in mice. The results suggest that glutaurine possesses a radiation protective effect in animals irradiated with LD50/30 of roentgen rays and 60Co gamma rays. The compound has a favourable effect also when administered after irradiation. Its protective effect is especially marked in case of prolonged irradiation. Among the combinations best results were obtained by its simultaneous administration with subminimal doses of AET or cystamine. Some of its derivatives also exhibited considerable protection against irradiation with roentgen rays. PMID- 6278853 TI - Radiation damage to the rat spinal cord. A radiologic model. AB - A new kinetic model for evaluation of the cumulative biologic effect of fractionated irradiation was described previously. Survival curves were calculated from two parametric Huggett formulae in agreement with the connection to NSD. The model has been further developed and now consists of 4 simple parameters based on radiation biologic considerations. PMID- 6278852 TI - Differences in the plasma kinetics of daunorubicin in normal and leukemic rats. AB - The plasma pharmacokinetics of the anthracycline drug daunorubicin in doses of 1 and 10 mg/kg body weight were compared in normal and leukemic inbred Sutton August rats (50-70 x 10(9) leukemic cells/1 blood). The area under the plasma concentration versus time curve was about 25 times larger in the leukemic rats at both dose levels. The data indicate that daunorubicin has a lower volume of distribution in leukemic rats as compared with normals. The large reduction in plasma clearance of daunorubicin in the leukemic rats can to some extent be explained by impairment of hepatic and renal functions but the results indicate that the leukemic state per se was of importance for the observed pharmacokinetic differences. This finding may be of importance for the treatment of patients with a high leukemic cell load, since higher retention of the drug in the central compartment leads to lower concentrations in poorly vascularized tissues, which favours the development of resistant leukemic cell clones. PMID- 6278854 TI - Effect of ionizing radiation on platelet function in vitro. AB - The effect of ionizing radiation on platelet function was investigated in vitro. Platelet-rich plasma (300 x 10(9/1)) was irradiated with doses of 1, 4, 10, 20 and 50 Gy. Platelet function tests were performed on both irradiated and control (non-irradiated) platelet samples. The platelet function tests were (1) platelet aggregation by ADP (1, 2, 4 mumol final concentration), adrenaline and collagen, (2) ADP-release from platelets, (3) clot retraction and (4) platelet factor-3 availability. It was found that roentgen irradiation of platelets in vitro did not affect these platelet functions tests. PMID- 6278855 TI - Disturbances of fertility in female mice 99Sr-contaminated as foetuses. AB - Pregnant CBA-mice were intravenously injected with 90Sr doses of between 46 and 740 kBq per animal. Their female descendants were mated to untreated CBA-males during a period of 7 months and the reproductive capacity was investigated, i.e. the number of litters born as well as the number of young per female. At the end of the mating period the females were killed and their ovaries microscopically analysed regarding the number of remaining oocytes and follicles. A presumptive correlation between the manifested decrease of the reproductive capacity and the microscopically observed injury was analysed. The fertile conditions of the in utero treated females seemed not affected by 90Sr doses below 370 kBq per dam, although the microscopic examination revealed that 90Sr amounts of 46.3 kBq reduced the total number of oocytes and follicles by one third. PMID- 6278856 TI - Acute gastric erosions in the rat. I. Microangiography. AB - Microangiography of the gastric mucosa was performed in 16 adult rats 5 hours after administration of indomethacin. Superficial gastric erosions were induced irrespective of the route of administration. Microangiography demonstrated absence of vascular dilatation underneath the superficial gastric erosions, suggesting absence of protracted vascular spasm or microthrombosis in eroded areas. The presence of normal microangiographic appearances underneath the gastric erosions suggests that such lesions may be induced by mechanisms other than by vascular injury. PMID- 6278857 TI - Adrenocortical function compared with computed tomography of the adrenals in small cell carcinoma of the lung. AB - Autopsy in patients with pulmonary small cell carcinoma frequently reveals metastases to the adrenal glands. In order to test the hypothesis that CT scanning might demonstrate such metastases at the time of clinical diagnosis of the pulmonary disease, the results of adrenal CT in 15 patients were correlated to adrenocortical function tests. No statistically significant relationship was found. It is concluded that no reliable non-invasive method exists to diagnose adrenal metastases in patients with pulmonary small cell carcinoma. PMID- 6278859 TI - Alteration of gallium distribution in the rat using periodically related elements: enhanced early imaging. AB - Complexes of carrier-gallium and indium were administered to normal rats and rats with tumors 2 to 6 hours after an injection of carrier-free 67Ga citrate in order to determine their effect on the distribution of 67Ga. Both Ga and In rapidly and significantly decreased the blood activity. Imaging with the same dosage schedule showed decreased activity in soft tissue and viscera, and increased activity in bone, kidney, and bladder. In animals with tumor high doses of carrier-Ga were shown to deplete tumor activity as well as activity in viscera and soft tissues. Moderate doses of carrier-Ga allowed blood and soft tissue clearing with no significant loss of tumor activity. When In citrate was administered to animals with tumors 2 hours after administration of 67Ga, tumor activity continued to increase while soft tissue and visceral activity decreased. Simultaneous injection of In citrate and 67Ga drastically altered the distribution of 67Ga. Two hours after the injection activity was present only in the tumor, kidney, bladder, and bone. This rapid clearance of non-productive Ga brings forth the potential for use of short lived, positron emitting 68Ga coupled with emission tomography. PMID- 6278858 TI - Subcellular binding of 99Tcm-gluconate in infarcted myocardium in dogs. AB - Binding of 99Tcm-gluconate on the subcellular level in myocardial infarct tissue in the dog was investigated by means of a fractionated centrifugation technique. High isotope activity/g protein was recorded in the mitochondrial fraction indicating the 99Tcm-gluconate binds preferentially to a protein structure localized in the mitochondria. PMID- 6278860 TI - Comparison of cell proliferation in the regenerating bone marrow and spleen of irradiated mice with the use of 125I-iododeoxyuridine. AB - The method of 125IUdR labelling of hemopoietic cell populations was used to estimate differences of repopulation kinetics in the femoral bone marrow and spleen of sublethally irradiated mice. In regenerating bone marrow the cellularity and proliferation index, as measured by 6 h 125IUdR incorporation, were found to be inversely related. In the spleen the proliferation index increased parallelly with the increase in cellularity and indicates an ineffectiveness of a change in the set point of local cellularity control. Experiments with measuring the retention of 125IUdR in hemopoietic organs revealed, in the spleen of irradiated animals, phases of increased cell losses as well as immigration of labelled cells into this organ. PMID- 6278862 TI - Calibration and use of plane-parallel ionization chambers for the determination of absorbed dose in electron beams. AB - The dosimetry procedures to be used at electron energies, E0, below 15 MeV in order to determine absorbed dose to water with an ionization chamber have been investigated. The performance of different types of plane parallel ionization chambers was tested. A chamber suitable for use in accordance with the Nordic dosimetry recommendations was designed. Correction factors to be applied in order to use solid phantom materials instead of water were determined. PMID- 6278861 TI - Human serum albumin labelled with 99Tcm and 131I for total blood volume determination: A critical and comparative analysis in vivo and in vitro. AB - The increased plasma volume or total blood volume determined with radioactive tracers often have no physical or pathologic explanation. The plasma volume and blood dilution were simultaneously measured in 5 patients who had received human serum albumin labelled with 99Tcm or 131I. In each case and for each tracer, a biexponential dilution curve was obtained. The measured plasma volume was consistently higher after 99Tcm-labelled albumin. The results are correlated with the presence of non-protein artifacts arising from the labelling process and proteinaceous compounds arising from denaturation. PMID- 6278863 TI - Electron beams with mean energies at the phantom surface below 15 MeV. Supplement to the recommendations by the Nordic Association of Clinical Physics (NACP) 1980. PMID- 6278864 TI - [Intravascular papillary endothelial hyperplasia (HEVI of Masson)]. PMID- 6278865 TI - Comparison of pathological and biochemical studies of transplanted Morris hepatomas. PMID- 6278866 TI - Interrelationship between glycolysis and the anaerobic part of the pentose phosphate pathway of carbohydrate metabolism in the myocardium. PMID- 6278867 TI - Biochemical markers as indicators of prognosis in acute nonlymphocytic leukemia (ANLL). PMID- 6278868 TI - Polycyclic hydrocarbon metabolism: a plethora of phenomena. PMID- 6278870 TI - The distribution of 5-methylcytosine in guinea pig satellite DNA's as probed by the restriction enzymes Msp I and Hpa II. PMID- 6278869 TI - Intracellular Ca2+-dependent protease (calpain) and its high-molecular-weight endogenous inhibitor (calpastatin). PMID- 6278871 TI - Protein methylation: cytochrome c methylation as a model system. PMID- 6278872 TI - The structure and behavior of coronavirus A59 glycoproteins. PMID- 6278874 TI - Assembly of mouse hepatitis virus strain JHM. PMID- 6278873 TI - Relatedness of virion and intracellular proteins of the murine coronaviruses JHM and A59. PMID- 6278875 TI - Glycoprotein E1 of coronavirus A59: a new type of viral glycoprotein. PMID- 6278876 TI - Analysis of the functions of coronavirus glycoproteins by differential inhibition of synthesis with tunicamycin. PMID- 6278877 TI - Oligonucleotide fingerprinting of the RNA of different strains of infectious bronchitis virus. AB - 11 distinct oligonucleotide fingerprints were obtained in studies of the RNA of 13 isolates of IBV. Different serotypes had distinct fingerprints, but so did varieties within a serotype; allowing a greater degree of strain differentiation than was previously possible. Some conclusions can be drawn from the fingerprints concerning theories of origin and spread of IBV. PMID- 6278878 TI - Coronavirus cell-associated RNA-dependent RNA polymerase. PMID- 6278879 TI - On enteropathogenic bovine coronavirus. PMID- 6278880 TI - Polypeptide structure of bovine enteritic coronavirus: comparison between a wild strain purified from feces and a HRT 18 cell adapted strain. PMID- 6278882 TI - Intracellular protein synthesis and the in vitro translation of coronavirus JHM mRNA. PMID- 6278883 TI - Messenger RNAs of mouse hepatitis virus A59: isolation and characterization, translation in Xenopus laevis oocytes of RNAs 3, 6 and 7, UV target sizes of the transcription templates. PMID- 6278881 TI - The avian coronavirus multiplication strategy. PMID- 6278884 TI - Murine coronavirus RNA. PMID- 6278885 TI - Comparison of the RNAs of murine and human coronaviruses. PMID- 6278886 TI - In vivo and in vitro models of demyelinating diseases. PMID- 6278887 TI - Evolution of a coronavirus during persistent infection in vitro. PMID- 6278888 TI - Persistent infection with mouse hepatitis virus, JHM strain in DBT cell culture. AB - After inoculation with JHM strain into DBT cell monolayers, a persistently infected DBT cell culture was established without producing typical cytopathic changes after about 15th passages. By immunofluorescence virus specific antigen was demonstrated in 10 to 15% DBT cells. This persistently infected culture (JHM CC) was resistant to superinfection with parental JHM, but such resistance was not shown against vesicular stomatitis virus. JHM-CC virus produced small plaques on DBT cell monolayers. Temperature sensitive (TS) mutant, defective interfering (DI) particle or interferon was not detected in the JHM-CC. To intracerebral inoculation with JHM-CC virus, cortisone treated ICR mice survived without showing clinical signs, however, demyelinating lesions were produced in the brain and spinal cord of them. PMID- 6278889 TI - JHM infections in rats as a model for acute and subacute demyelinating disease. AB - An animal model with different central nervous system (CNS) disease processes associated with demyelination is described which provides a basis to analyse the pathogenetic mechanisms leading to these disorders. Intracerebral infection of rats with the murine coronavirus strain JHM can result in an acute encephalomyelitis with a short incubation period or in subacute to chronic encephalomyelitis occurring after prolonged incubation. The most prominent finding of the latter two diseases consists of typical demyelinated lesions distributed in selected areas of the CNS. The induction of high rates of animals with demyelination depends both on properties of the virus used for infection and host factors such as age and immune status. A high number of rats with demyelination was obtained by intracerebral inoculation of temperature sensitive mutants into suckling rats with maternal JHM antibodies. PMID- 6278890 TI - Host and virus factors associated with CNS cellular tropism leading to encephalomyelitis or demyelination induced by the JHM strain of mouse hepatitis virus. PMID- 6278891 TI - Pathogenicity and persistence of mouse hepatitis virus in inbred strains of mice. PMID- 6278892 TI - Influence of the immune system on the course of infection with murine coronavirus JHM in suckling mice. AB - The course of infection with murine corona virus JHM in C3H mice depends on the age of the animals. Mice up to 20 days of age are fully susceptible while mice older than 23 days resist the infection. Protection of suckling mice from death of infection can be provided by intraperitoneal administration of immune spleen cells but not by non-immune spleen cells from adult mice. The immune spleen cells can be generated by priming adult mice, or by priming non-immune spleen cells from adult mice in baby mice with inactivated JHM virus. Thus the immune system might well be involved in the different outcome of infection with JHM-virus in suckling and adult mice, but it does not seem to be the exclusive factor responsible for the achievement of natural resistance. PMID- 6278893 TI - Macrophages and resistance to JHM virus CNS infection. AB - Thioglycollate elicited peritoneal exudate cells from resistant SJL mice, younger susceptible SJL mice, and susceptible ASW, BALB/c, and C57/BL6 mice all exhibit extrinsic antiviral activity. The active cell was characterized as a Thy 1.2 negative, Ia negative, radiation resistant adherent cell. The antiviral activity was not due to nonspecific cellular cytotoxicity directed against the susceptible cell nor interferon. Adherent PE cells from resistant and susceptible SJL mice were similar with respect to the number of phagocytes, nonspecific esterase containing, Fc, and C3b receptor bearing cells. Finally, extrinsic antiviral activity was not dependent upon intrinsic antiviral activity. PMID- 6278894 TI - Vomiting and wasting disease, a coronavirus infection of pigs. PMID- 6278895 TI - Inborn resistance of mice to mouse hepatitis virus type 3 (MHV3): liver parenchymal cells express phenotype in culture. AB - Primary monolayer cultures of hepatocytes isolated from adult resistant A/J or partially resistant A/Sn or C3H/HeJ mice exhibited resistance to MHV3 as the respective macrophages do: Compared to susceptible C57BL/6 hepatocyte cultures, cytopathic effect occurred later and was restricted to small foci, coinciding with areas specifically labelled by immunofluorescence. Production of infectious particles was delayed, titers being 100 to 1000 fold lower at the moment of maximal yields in susceptible cultures. Pretreatment with interferon could reduce the titers in susceptible cultures to a level as seen in resistant cultures not treated with interferon. Nevertheless, interferon was not responsible for the genetic resistance of hepatocytes: it reduced virus titers in susceptible and resistant cultures to the same extent and the addition of specific antibodies to interferon after infection did not augment susceptibility of resistant cultures. We assume that intrinsic resistance of liver parenchymal cells is an important facet of inborn resistance of mice in vivo. PMID- 6278896 TI - Biology of coronaviruses 1980. PMID- 6278897 TI - Genome structure of mouse hepatitis virus: comparative analysis by oligonucleotide mapping. AB - Several natural variants of mouse hepatitis viruses have been compared by the T1 oligonucleotide fingerprinting technique. In general, they have diverged quite extensively. However, MHV-3, a hepatotropic strain, and A59, a nonpathogenic strain, were found to be extremely related. Yet, each of them contains 2-4 specific oligonucleotides. One of the MHV-3-specific oligonucleotides was mapped in 30S poly(A)-containing RNA or 6-7 Kb from the 3-end and the other near the 5' end of the genome. These two genetic regions might be associated with viral pathogenicity. In addition, two JHM plaque variants, DL producing large plaques and DS producing small plaques, were also compared. They share almost all T1 oligonucleotides, but each contains one unique spot. The DL-specific oligonucleotide was mapped in 21S poly(A)-containing RNA or at about 4 kb from the 3'-end. Finally, the MHV genome was found to contain the "cap" structure, confirming that it is a positive-stranded RNA. PMID- 6278898 TI - The structure of the canine coronavirus. PMID- 6278899 TI - The effects of relaxin on cyclic-AMP an ornithine decarboxylase levels in target tissues. PMID- 6278900 TI - Relaxin receptors and a study of the physiological roles of relaxin. PMID- 6278901 TI - Target tissues for relaxin. PMID- 6278902 TI - The nutritional significance, metabolism, and function of myo-inositol and phosphatidylinositol in health and disease. AB - Recent advances in nutritional and biochemical research have substantiated the importance of inositol as a dietary and cellular constituent. The processes involved in the metabolism of inositol and its derivatives in mammalian tissues have been characterized both in vivo and at the enzyme level. Biochemical functions elucidated for phosphatidylinositol in biological membranes include the mediation of cellular responses to external stimuli, nerve transmission, and the regulation of enzyme activity through specific interactions with various proteins. Inositol deficiency in animals has been shown to produce an accumulation of triglyceride in liver, intestinal lipodystrophy, and other abnormalities. The metabolic mechanisms giving rise to these latter phenomena have been extensively studied as a function of dietary inositol. Altered metabolism of inositol has been documented in patients with diabetes mellitus, chronic renal failure, galactosemia, and multiple sclerosis. A moderate increase in plasma and nerve inositol levels by dietary supplementation has been suggested as a means of treating diabetic neuropathy, although excessively high levels, such as are found in uremic patients, may be neurotoxic. A thorough consideration of the biochemical functions of inositol and a further characterization of various diseases with the aid of appropriate animal models may suggest a possible role for inositol and other dietary components in their prevention and treatment PMID- 6278903 TI - Angulated fluoroscopy for needle biopsy localization. PMID- 6278904 TI - Prophylaxis for hepatitis A. AB - Hepatitis A is a common communicable disease in this country. Each diagnosed case of hepatitis A involves the problem of selecting contacts for post-exposure immunoglobulin. These decisions should be systematic, based on current knowledge of epidemiology, kinetics of viral excretion by infected persons and the influence of time on the efficacy of immunoglobulin prophylaxis. An algorithm incorporates this information into a strategy consistent with the United States Public Health Service guidelines. PMID- 6278905 TI - Prediction of sustained antihypertensive efficacy of chronic captopril therapy: relationships to immediate blood pressure response and control plasma renin activity. AB - The blood pressure (BP) lowering effect of the orally active angiotensin converting enzyme inhibitor, captopril (SQ14225), was studied in 59 hypertensive patients maintained on a constant sodium intake. Within 2 hours of the first dose of captopril BP fell from 171/107 to a maximum low of 142/92 mm Hg (p less than 0.001), and after 4 to 8 days to treatment BP averaged 145/94 mm Hg (p less than 0.001). The magnitude of BP drop induced by captopril was significantly correlated to baseline plasma renin activity (PRA) both during the acute phase (r = -0.38, p less than 0.01) and after the 4 to 8-day interval (r = -0.33, p less than 0.01). Because of considerable scatter in individual data, renin profiling was not precisely predictive of the immediate or delayed BP response of separate patients. However, the BP levels achieved following the initial dose of captopril were closely correlated to BP measured after 4 to 8 days of therapy, and appeared to have greater predictive value than control PRA of the long-term efficacy of chronic captopril therapy despite marked BP changes occurring in some patients during the intermediate period. Because of these intermediate BP changes, addition of a diuretic to enhance antihypertensive effectiveness of angiotensin blockade should be restrained for several days after initiation of captopril therapy. PMID- 6278907 TI - Diagnostic considerations in cardiomyopathy: unique scintigraphic pattern of diffuse biventricular technetium-99m-pyrophosphate uptake in amyloid heart disease. PMID- 6278906 TI - Value of positive myocardial technetium-99m-pyrophosphate scintigraphy in the noninvasive diagnosis of cardiac amyloidosis. AB - Ten consecutive patients with tissue-proven amyloidosis, seven of whom presented with congestive heart failure, were found to exhibit intense diffuse uptake of technetium-99m-pyrophosphate (Tc-99m-PYP) on cardiac radionuclide imaging. The patients exhibited echocardiographic and systolic time interval abnormalities suggesting combined restrictive and congestive cardiomyopathic changes. On M-mode echocardiograms, there was symmetrically increased thickness of the interventricular septum and left ventricular (LV) posterior wall in diastole (10 of 10), decreased fractional shortening of the LV minor axis diameter in systole (eight of nine), and decreased percent thickening of the LV minor axis diameter in systole (eight of nine) and LV posterior wall (10 of 10) in systole. Three patients demonstrated enlarged LV end-diastolic diameter. All 10 patients had abnormal PEP/LVET and eight had shortened LVETI. When combined with noninvasive tests of LV performance, positive myocardial pyrophosphate (PYP) scanning provides a new and useful adjunct in the diagnosis of amyloid heart disease. PMID- 6278908 TI - Noninvasive diagnosis of cardiac amyloidosis by technetium-99m-pyrophosphate myocardial scintigraphy. PMID- 6278909 TI - Balkan nephritis and adenovirus infection. PMID- 6278910 TI - Role of calcium-blocking agents in treatment of cardiac arrhythmias related to myocardial ischemia. AB - Potential antiarrhythmic effects of calcium (slow-channel)-blocking drugs ca be indirect or direct. The indirect effect would affect ischemia-induced arrhythmias by improving myocardial perfusion or decreasing myocardial oxygen consumption. The direct effect would affect the electrical activity of the sinoatrial (SA) and atrioventricular (AV) nodes and any type of cardiac fibers depolarized to a level at which the slow inward current becomes activated. Although myocardial ischemia could become a setting of slow-channel-dependent conduction or automaticity, or both, there is no conclusive evidence that slow-channel-blocking drugs suppress ischemia-induced ventricular arrhythmias. Thus the principal usefulness of slow channel-blocking drugs may be expected in the treatment of supraventricular arrhythmias and in the prevention of arrhythmogenic ischemia. PMID- 6278911 TI - Hypoxia, calcium fluxes, and inotropic state: studies in cultured heart cells. AB - Methods have been developed for the study of cellular mechanisms underlying a variety of physiologic and pathophysiologic phenomena using cultured chick embryo heart cells. These spontaneously beating monolayer preparations lack appreciable diffusion barriers, and interstitial space components can be defined and controlled conveniently. These properties facilitate the measurement of transmembrane fluxes and cellular contents of sodium, potassium, and calcium. Concomitantly, methods have been developed to measure the contractile state as reflected in the amplitude and velocity of movement of individual myocytes in the culture. Experiments are described in which the contractile responses of cultured heart cells to graded hypoxia demonstrated a critical PO2 level around 12 mm Hg, below which progressive impairment of contractile function occurred. In preparations with iodoacetate-induced glycolytic blockade, enhanced sensitivity to hypoxia occurred. Although experiments are described, demonstrating the applicability of this model system to the study of problems including the mechanism of cardiac glycoside-induced increases in inotropic state and the definition of calcium flux pathways important in excitation-contraction coupling. PMID- 6278912 TI - Sympathomimetic amines: potential clinical applications in ischemic heart disease. AB - Sympathomimetic amines are useful in the treatment of patients with ischemic heart disease complicated by heart failure and shock. These agents influence the cardiovascular system by action on alpha-adrenergic, beta-adrenergic, and dopamine receptors. Recent evidence has demonstrated the existence of subtypes of the classic adrenergic and dopamine receptors that mediate distinct physiologic effects. The relative actions of sympathomimetic amines on these receptors differ substantially, resulting in considerable variation in their cardiac and peripheral vascular effects. Two classes of sympathomimetic amines are being intensively investigated at present: (1) compounds acting predominantly on beta 1 adrenergic receptors (i.e., they increase cardiac contractile force with little or no peripheral vascular effects) and (2) compounds acting on both beta 1 adrenergic and dopamine receptors. Orally active compounds of these two classes have been synthesized recently and are now under study for the treatment of patients with heart failure. Results of preliminary studies with such components are briefly reviewed. PMID- 6278913 TI - Absence of slow channel-dependent conduction within the His-Purkinje (bundle branch) reentrant circuit: a clinical and experimental study of the effects of verapamil. PMID- 6278915 TI - Overdrive suppression of atrial pacemaker tissues in the alert, awake dog before and chronically after excision of the sinoatrial node. PMID- 6278914 TI - Prevention of propranolol withdrawal mechanism by prolonged small dose propranolol schedule. AB - Abrupt withdrawal of propranolol may be followed by a "propranolol withdrawal syndrome "due, at least in part, to enhanced beta adrenergic sensitivity. Tapering propranolol dosage is frequently used in the hope of preventing adverse withdrawal events but the success of such a maneuver has not been shown. The rationale for the dose tapering schedule in this study was based on earlier observations after abrupt withdrawal of propranolol. Nine hypertensive patients were gradually withdrawn from long-term propranolol therapy, either by serial dose reduction for 6 to 9 days (n = 3) or by reduction to a prolonged small dose (30 mg daily) for 2 weeks before complete withdrawal (n = 6). During dose reduction of propranolol and for 2 additional weeks of placebo therapy, serial measurements were made of cardiac sensitivity to isoproterenol, heart rate at rest, blood pressure, plasma catecholamines, serum thyroxine (T4) and triiodothyronine (T3) and symptoms. Serial dose reduction decreased but did not prevent cardiac hypersensitivity in two of three patients. The prolonged small dose therapy largely prevented cardiac hypersensitivity and overshoot in heart rate, blood pressure and plasma catecholamines and symptoms. Serum T4 decreased significantly and T3 tended to increase during and after prolonged small dose treatment. These results indicate that prolonged administration of small dose propranolol before complete withdrawal in hypertensive patients prevents enhanced cardiac beta adrenergic sensitivity and other adverse events. PMID- 6278916 TI - Myocardial infarct imaging. PMID- 6278917 TI - Components of fiber bind iron in vitro. AB - Interactions among fiber, inorganic iron, and substances known to chelate iron were examined in vitro. Cellulose, neutral and acid fractions of lignin, psyllium mucilage, and pectin were incubated with 59Fe SO4 at various pH's, in various concentrations, and in the presence or absence of known chelators of iron including ascorbate, citrate, cysteine, fructose, and EDTA. Insoluble components of fiber were tested in a precipitation-centrifugation system, soluble components were tested by equilibrium dialysis. Lignins and psyllium mucilage had the greatest capacity to bind ferrous iron under conditions that approximated those of the proximal intestine postprandially; cellulose and pectin were less potent. Dissociation constants of binding were similar for several components, suggesting the existence of comparable binding sites in different components of fiber. Citrate and EDTA inhibited the binding of iron markedly but other chelators were much less effective. These reactions among fiber, inorganic iron, and other constituents of food may influence the bioavailability of dietary iron and the simple systems described here offer a means of dissecting interactions that are complex in vivo. PMID- 6278918 TI - Components of fiber impair iron absorption in the dog. AB - We have demonstrated previously that semipurified components of fiber bind ferrous iron in vitro. The present studies examined the possibility that these same fibers would decrease absorption of iron by the canine small bowel. Healthy and anemic dogs were studied chronically. Two preparations were needed; soluble components of fiber were tested by continuous perfusion of a duodenojejunal segment of approximately 100 cm, insoluble components were tested in Thiry-Vella fistulae of jejunum (25 cm). The results confirm that, 1) anemic dogs absorb iron more efficiently than do healthy animals, and 2) that there is a slight, but significant, decrease in iron absorption during the 4 to 6 h of an acute experiment. To obviate problems of interpretation because of the latter point, all studies featured a control period, before and after, the test period. Lignin and psyllium mucilage were potent inhibitors of iron absorption, pectin less so, and cellulose was without effect in this system. The results in vivo correlate well with the binding of iron in vitro. PMID- 6278919 TI - The effects of oral iodized oil on intelligence, thyroid status, and somatic growth in school-age children from an area of endemic goiter. AB - One hundred goitrous school children received 475 mg iodized oil by mouth, while 100 controls received mineral oil, on a double-blind basis. On follow-up 22 months later the urinary iodine had increased and goiter size had decreased in both groups, more strikingly in the iodine-treated children. There were no consistent differences between the two treatment groups in rate of somatic growth or performance on the Stanford-Binet and Bender tests. Because of the complexities introduced by increases in urinary iodine in the controls, we compared goiter reduction with improvement in IQ score in all children, regardless of group, and found a significant relationship (p = 0.014), particularly in girls (p = 0.029). We conclude that oral iodized oil is an attractive alternative to its injection but we recommend an approximate doubling of the dose used here for more effective control. Also, while our data are not conclusive, they support the possibility that correction of iodine deficiency may improve mental performance in school age children, particularly girls. PMID- 6278920 TI - Analysis of fiber components. PMID- 6278921 TI - A factor in food which impairs Na+-K+-ATPase in vitro. AB - Extracts of foods were examined to identify materials which inactivate Na+-K+ dependent ATPase. Active material which we call food factor was found in tea, cocoa, red wines, and in certain other dietary components. A unit of activity is defined and the material is compared with ouabain and an inhibitory flavone. The possible role of such a food factor in the regulation of sodium metabolism and in the causation of essential high blood pressure is discussed. PMID- 6278922 TI - No link between Poland syndrome and leukemia? PMID- 6278923 TI - Congenital thrombocytopenia, hepatosplenomegaly, and growth retardation. A clinicopathologic conference. PMID- 6278924 TI - Pertechnetate imaging following cimetidine administration in Meckel's diverticulum of the ileum. AB - The demonstration of secretory gastric mucosa in Meckel's diverticulum of the ileum in patients undergoing radionuclide imaging with 99mTc sodium pertechnetate is greatly improved with the prior administration of cimetidine. This results in enhanced accumulation of pertechnetate in the gastric mucosa along with reduced excretion of the radionuclide into the gastric lumen and, consequently, less intestinal radioactivity. PMID- 6278925 TI - Longitudinal studies of infectious diseases and physical growth of children in rural Bangladesh. II. Incidence of diarrhea and association with known pathogens. AB - Longitudinal studies were done in two villages rural Bangladesh to learn more about the interactions between infectious diseases and the nutritional status of children. Diarrheal diseases, identified by surveillance of 197 children aged 2 60 months, were studied for bacterial, viral and parasitic enteropathogens in 1978-1979. The annual incidence of diarrhea was highest in children aged 2-11 months, and declined progressively with age from seven to four episodes per child per year. An enteropathogen was identified from rectal cultures taken during diarrhea in 51% of episodes and from 6% of monthly cultures taken when diarrhea was not present. Enterotoxigenic Escherichia coli were the pathogens found most frequently, followed by shigellae and rotaviruses. Diarrheal episodes associated with shigellae had the longest duration, while episodes associated with Vibrio cholerae or with rotavirus were more frequently associated with dehydration. E. coli diarrhea had a peak incidence during the hot months, and shigellosis was more frequent during the cool, dry months. PMID- 6278926 TI - A large outbreak of hepatitis A in a day-care center: association with non-toilet trained children and persistence of IgM antibody to hepatitis A virus. AB - In early summer, 1979, a large outbreak of hepatitis A occurred in an Oklahoma day-care center. A total of 41 cases were confirmed, all in adults. Of the 115 non-employee households represented by children in the center, 19 households (16.5%) had one or more cases of hepatitis. Hepatitis occurred in 29% of the households with at least one non-toilet-trained child, compared to 2% of the households without such a child (p = 0.00004). At least four (15%) of 27 center employees had hepatitis. Of 26 cases tested serologically, all were positive for hepatitis A antibody (anti-HAV), and 24 of these 26 were also positive for anti HAV of immunoglobulin class M (anti-HAV IgM), at an average time after onset of illness of 80 days (range, 38-142 days). Three of ten persons remained anti-HAV IgM-positive 164 days after onset. PMID- 6278927 TI - Analysis of the genomes of bluetongue viruses recovered from different states of the United States and at different times. AB - The ribonucleic acid (RNA) genomes of the original representatives of the four serotypes of bluetongue virus (BTV) isolated from the United States (BTV-10, BTV 11, BTV-13 and BTV-17) have been analyzed by polyacrylamide gel electrophoresis. Ten double-stranded RNA species were resolved for each virus and categorized into the size classes of large (L segments #1-3), medium (M segments #4-6) and small (S segments #7-10). Minor electrophoretic mobility differences were observed for the M5 and S10 RNA segments of BTV-10 by comparison with the other US prototype viruses. The oligonucleotide fingerprints of the individual RNA species of BTV-11 were obtained and compared to each other providing proof that each of the viral double-stranded RNA species contains unique genetic information. Alternate isolates of BTV-11 isolated in the same year (1973), but from different states (Texas, Idaho, California and Oregon), or in different years (1963, 1970, 1974 and 1975), but from the same state (Colorado), have been analyzed by both RNA gel electrophoresis and L, M and S RNA oligonucleotide fingerprinting. When compared to the individual RNA fingerprints of the US prototype BTV-11 strain, the results indicated that there has been both genetic drift (accumulation of point mutations) and recombination (RNA segment reassortment) for BTV-11 in the United States over a 12-year period since 1963. PMID- 6278928 TI - Norwalk virus gastroenteritis following raw oyster consumption. AB - In January, 1980, six out of 13 persons (46%) attending a party in a small northwest Florida town near the Gulf of Mexico became ill with Norwalk virus gastroenteritis after eating raw oysters. Symptoms experienced by the ill persons were principally nausea (100%), vomiting (83%) and diarrhea (50%) and were of brief duration. The symptom complex and epidemiology of Norwalk virus infection closely resemble the gastrointestinal illness commonly referred to as the 24-hour intestinal flu or "stomach flu." Norwalk virus infection was identified in this outbreak by application of a recently developed sensitive and specific serologic radioimmunoassay. Oysters from the incriminated batch had fecal coliform levels above recommended standards; however, recent studies of oyster-harvesting waters have shown only a weak correlation between fecal coliforms and the presence of enteric viruses. Further studies are needed to determine whether modifications of monitoring modalities for oyster-harvesting waters are needed. PMID- 6278929 TI - Thalassemia: recent insights into molecular mechanisms. AB - Recent advances in defining the molecular basis for the thalassemia syndromes are discussed. We now realize that the causes of the thalassemia phenotype are diverse and include gene deletions, nuclear RNA processing defects, nonsense mutations, fusion genes, termination codon mutants, and unstable globin chains. PMID- 6278931 TI - Nursing care in childhood cancer: update. PMID- 6278930 TI - Albright's hereditary osteodystrophy: a review. PMID- 6278932 TI - Hypothalamic-pituitary-adrenocortical function during long-term low-dose dexamethasone therapy in hyperandrogenized women. AB - Hypothalamic-pituitary-adrenocortical (H-P-A) function was tested in 14 hyperandrogenized women, aged 17 to 32 years, who had been continuously treated with nightly single-dose oral dexamethasone, 0.25 to 1.00 mg, for 3.7 to 16.5 months. Daily AM serum cortisol concentrations were measured in nine subjects after discontinuation of dexamethasone. Basal cortisol concentrations returned to normal (greater than or equal to 6.0 micrograms/dl) within 36 hours in 67%, within 60 hours in 89%, and within 84 hours in 100%. Median time to return to normal was between 12 and 36 hours. Rate of return correlated with both the dose adjusted duration of dexamethasone therapy (p less than 0.01) and the degree of adrenocortical suppression during treatment (p less than 0.01). The H-P-A response to insulin-induced hypoglycemia and the adrenal response to an acute intravenous adrenocorticotropic hormone (ACTH) challenge were evaluated in eight subjects 12 to 36 hours after the final dexamethasone dose. Thirty-eight percent demonstrated normal cortisol increments to hypoglycemia, 25% had blunted or absent cortisol responses to hypoglycemia but normal cortisol increments to exogenous ACTH, and 38% had blunted or absent responses to both hypoglycemia and exogenous ACTH. The responsiveness of the H-P-A axis correlated with the degree of adrenocortical suppression (p less than 0.05) but not with dose of dexamethasone or duration of treatment. Small doses of dexamethasone are not necessarily "physiologic", but dexamethasone therapy with maintenance of serum cortisol levels greater than or equal to 2.0 micrograms/dl was associated with rapid return to normal basal cortisol concentration and a normal cortisol response to insulin-induced hypoglycemia. PMID- 6278933 TI - Actions of progestins on the morphology and biochemistry of the endometrium of postmenopausal women receiving low-dose estrogen therapy. AB - Endometrial biopsies were obtained from postmenopausal women receiving 0.625 mg Premarin daily and either 2.5 or 5 mg norethindrone daily or 150 or 500 microgram dl-norgestrel daily for 10 days each month. Sample were taken during the estrogen-only phase of treatment and on the sixth day of combined estrogen/progestin administration. Progestin exposure caused marked morphologic and biochemical changes as well as features comparable with the premenopausal luteal phase. Thus, progestins oppose the stimulation of premarin to the postmenopausal endometrium. However, the currently recommended dosage of norethindrone and dl-norgestrel greatly exceed those necessary to suppress endometrial proliferation effectively. The recommended daily dosage may be lowered without losing protective effect. This reduction will probably minimize the risk of dose-dependent progestin side effects. PMID- 6278934 TI - Leber's idiopathic stellate retinopathy. AB - Three patients with Leber's idiopathic stellate retinopathy-a 16-year-old girl, a 26-year-old man, and a 7-year-old girl-also had acute multifocal chorioretinitis. The second patient also had an associated idiopathic peripheral seventh nerve palsy. Extensive neurologic testing and viral cultures gave no explanation of the origin of the retinopathy. However, each of the patients had had an upper respiratory or gastrointestinal illness shortly before the onset of the condition, suggesting an infectious origin. In all three patients the optic disk swelling resolved spontaneously and the stellate exudate gradually faded. PMID- 6278935 TI - Human pancreatic adenocarcinoma line Capan-1 in tissue culture and the nude mouse: morphologic, biologic, and biochemical characteristics. AB - Human pancreatic ductal adenocarcinoma line Capan-1 was studied in tissue culture and the nude mouse. In tissue culture, the neoplastic cells grew as large epithelial-like mucin-producing cells. Subcutaneous and intraperitoneal transplantation of neoplastic cells into nude mice resulted in tumor formation characterized by marked invasiveness and distant metastases. Histologically, the tumor appeared as a well-differentiated mucin-producing adenocarcinoma morphologically resembling the tumor of origin. Chromosomal analysis showed a human karyotype with a chromosome number between 51-61. Lactate dehydrogenase and beta 2-microglobulin used as tumor markers were present in both tissue culture and the serum of tumor-bearing mice. The neoplasm, which was characterized by an increased level of cAMP, had lost completely the ability to respond to secretin stimulation. The tumor grown in the nude mouse was resistant to treatment with 5 fluorouracil, behavior identical to that of the original tumor. Diphtheria toxin resulted in complete tumor destruction. Because Capan-1 tumor grown in the nude mouse shows morphologic, biologic, and biochemical characteristics similar to the tumor of origin, it may be an invaluable tool in furthering understanding of the biology of human pancreatic cancer. PMID- 6278936 TI - S-100 protein in soft-tissue tumors derived from Schwann cells and melanocytes. AB - In soft tissues outside the central nervous system, S-100 protein is found normally only in Schwann cells. Using the peroxidase-antiperoxidase immunohistochemical method S-100 was also found in tumors derived from Schwann cells and melanocytes, including neurofibromas, neurilemomas, granular cell myoblastomas, cutaneous nevi, and malignant melanomas. S-100 was not detected in malignant Schwannomas, neuroblastomas, oat cell carcinomas, medullary carcinomas of the thyroid, paragangliomas, or meningiomas. S-100 was also absent from neoplasms of soft tissues not usually considered to arise from cells of neural crest origin. S-100 appears to be a useful marker for identifying neoplasms derived from Schwann cells and melanocytes. PMID- 6278937 TI - Animal model of human disease. Skeletal hyperostoses: viral induction of avian osteopetrosis. PMID- 6278938 TI - Multiple cellular forms of corticotrophs in surgically removed pituitary adenomas and periadenomatous tissue in Cushing's disease. AB - Transsphenoidally removed samples of pituitary adenomas from 14 patients with Cushing's disease and 5 patients with Nelson's syndrome always contained groups of uniform small ACTH-cells. Antibodies against the pro-opiocortin precursor fragments beta-endorphin, ACTH, and 16k-peptide recognized material in typical adenoma cells. A subpopulation of these cells, varying in number from sample to sample, specifically exhibited alpha-melanotropin immunoreactivity. Most periadenomatous samples showed signs of severe degeneration. Typical Crooke cells only occurred in samples from patients with Cushing's syndrome, but, with this exception, no clear differences between pituitaries of patients with Cushing's and Nelson's syndromes could be discerned. Two other forms of ACTH-immunoreactive cells were observed: rare, single, highly immunoreactive cells, with characteristics of both normal and Crooke cells, and numerous syncytial groups of cells in an advanced state of disintegration, presumably the remnants of hyperplastic follicles. The four different corticotrophs are characterized according to their fine structure and immunoreactivity in this study. PMID- 6278939 TI - Brain phagocytes: source of high acid phosphatase activity. AB - A stab wound was produced in the adult rat cerebral cortex, and the progress of enzyme cytochemistry of phagocytosis was studied over 450 days. Light- and electron-microscopic observations were made to establish the origin of high acid phosphatase activity commonly seen at the site of brain lesions. Cells with phagocytic potential became differentiated and activated by the presence of degenerating neurons. The Golgi-lysosomal system of the phagocytes became elaborated, as evidenced by thiamine pyrophosphatase and acid phosphatase activities, the synthesis of acid phosphatase was increased, and the enzyme then secreted into the digestive vacuoles containing dead cells to be digested. Progress of the digestive process resulted in the accumulation of large amounts of acid phosphatase reaction product within the digestive vacuoles. The results showed that the phagocytes were the only detectable source of increased acid phosphatase activity at the site of injury in the cerebral cortex. In contrast to the phagocytes, newly formed multi-nucleated giant cells exhibited weak acid phosphatase, and intense cytochrome oxidase activities, the difference between the two cells reflecting the functional characteristics of each. PMID- 6278940 TI - Comparison of canine corticosteroid responses to mean and phasic increases in ACTH. AB - To determine the dynamics and magnitudes of adrenal corticosteroid responses to ACTH, we measured arterial plasma ACTH and corticosteroid concentrations in conscious dogs during infusions of ACTH or saline. Synthetic alpha 1-24-ACTH was infused at rates of 300,900, or 4,500 ng/30 min either as constant infusions or as three equal short infusions at 10-min intervals. In dogs infused with saline, plasma ACTH fluctuated, whereas corticosteroids did not, suggesting that ACTH is secreted episodically in dogs as in man. The magnitudes of the plasma corticosteroid responses to ACTH infusions were linearly related to the logarithm of the total amount of ACTH infused in 30 min and not to the pattern of administration. In all ACTH infusion experiments, the lag between an increase in arterial ACTH and corticosteroids was not less than 3 min. Mean ACTH half disappearance time, metabolic clearance rate, and volume of distribution estimated from the different experiments ranged between 1.8 and 2.1 min, 24 and 38 ml . kg-1 . min-1, and 95 and 114 ml/kg, respectively. Collectively, these results explain the apparent paradox that corticosteroid responses to ACTH releasing stimuli can be initiated before a detectable increase in ACTH above the highest control value (Wood et al. Apparent dissociation of ACTH and corticosteroid responses to ml/kg hemorrhage in conscious dogs. Endocrinology In press). PMID- 6278941 TI - Interrelationships of ion transport in rat submaxillary duct epithelium. AB - The transport of Na+, K+, Cl-, and HCO3(-) across the epithelium of the rat submaxillary salivary duct is postulated to be due to the coupling of the basolateral Na+-K+-ATPase with various ion transport systems in the luminal and basolateral membranes. Na+ reabsorption depends on the presence of a rheogenic (Na+ conductance) and an electroneutral (Na+:H exchange) pathway, both of which are sensitive to amiloride. K+ secretion is postulated to be mediated by a K+: H+ antiport, coupling between Na+ reabsorption and K+ secretion, thus depending on local H+ ion concentration. The ratio between electroneutral Na+ influx and K+ efflux, therefore, determines the rate of HCO3(-) secretion. In the absence of Na+ influx, although K+ efflux falls, HCO3(-) secretion rises to a value equal to that of K+ secretion. The maintenance of K+ secretion in the absence of luminal Na+ requires an additional Na+-entry step across the basolateral membrane, also postulated to be due to Na+:H+ exchange. PMID- 6278942 TI - Cyclic nucleotide antagonists of cholecystokinin: structural requirements for interaction with the cholecystokinin receptor. AB - Previously, we have found that, in pancreatic acini, butyryl derivatives of cGMP antagonize the action of cholecystokinin by inhibiting binding of the peptide to its membrane receptors. In the present study, we found that derivatives of cAMP and cIMP can also inhibit binding of cholecystokinin as well as its actions on acinar cell function. Moreover, the inhibition caused by cyclic nucleotide derivatives did not require the presence of a butyryl moiety, because certain 8 bromo-cyclic nucleotides also inhibited the interaction of cholecystokinin with its receptors. Cyclic nucleotide derivatives can also increase pancreatic enzyme secretion; however, for the various cyclic nucleotides tested, there was no apparent correlation between their abilities to stimulate enzyme secretion and their abilities to antagonize the actions of cholecystokinin. Finally, cyclic nucleotide derivatives also inhibited binding of 125I-cholecystokinin to antibodies that were specific for the biologically active, C-terminal region of cholecystokinin. Thus, certain cyclic nucleotide derivatives possess a conformational structural which resembles that of the biologically active portion of cholecystokinin, and this structural similarity accounts for the abilities of these nucleotide derivatives to interact with cholecystokinin receptors and, by so doing, to inhibit the action of cholecystokinin on its target tissues. PMID- 6278943 TI - Intestinal phospholipase A and triglyceride lipase: localization and effect of fasting. AB - We investigated the distribution of phospholipase A and triglyceride lipase in the rat small intestine and the effects of heparin and hormones on enzyme release. Phospholipase A activity was 10 times higher in the ileum than in the jejunum; triglyceride lipase activity was threefold higher in the jejunum than in the ileum. Activities of both enzymes were much greater in villus than in crypt cells. The specific activity of phospholipase A was highest in microsomes and least in cytosol. The crude nuclei and brush-border fraction contained 40.5% of total phospholipase A activity; mitochondria contained 33.8%; and microsomes, 17.4%. Phospholipase A activity increased significantly in the distal intestinal mucosa in fasted rats compared with controls. Heparin did not increase the release of phospholipase A by isolated intestinal cells or perfused intestinal vasculature. Thus, the small intestine probably does not contribute significantly to the phospholipase A activity of postheparin plasma. Hormones and cAMP, which inhibit the secretion of phospholipase A and triglyceride lipase from isolated hepatocytes, had no effect on the release of either enzyme from intestinal cells. PMID- 6278944 TI - beta-Adrenergic relaxation and cAMP kinase activation in coronary arterial smooth muscle. AB - If beta-adrenergic relaxation of smooth muscle is partly mediated by the adenosine 3',5'-cyclic monophosphate (cAMP) system, then beta-stimulation should be correlated to activation of cAMP-dependent protein kinase (cPK). Studies were performed with bovine coronary arterial strips to identify isozymic forms of cPK and to determine if beta-relaxation is correlated to activation of cPK (reflected by elevated ratios of cPK activity without cAMP to cPK activity with cAMP). Both ion exchange chromatography and a new electrophoretic technique revealed two cPK isozymes (types I and II). No change in cPK activity occurred in strips contracted with 30 mM KCl. In contrast, dose- and time-dependent relaxation during beta-stimulation with isoproterenol was highly correlated to parallel increases in cPK activity. Increased cPK activity was inhibited in assays performed with a specific inhibitor of cPK. Both relaxation and activation of cPK were abolished during beta-adrenergic blockade with propranolol. Relaxation by KCl removal or the ionophore R02-2985, unlike beta-mediated relaxation, did not increase cPK activity. These findings show that beta-mediated relaxation of isolated coronary arterial strips specifically activates cPK, and they support the hypothesis that beta-induced relaxation of vascular smooth muscle involves the cAMP system. PMID- 6278945 TI - Sympathetic transients caused by abrupt alterations of carotid baroreceptor activity in humans. AB - We examined the role of carotid baroreceptors in the short-term modulation of sympathetic outflow to the muscle vascular bed and parasympathetic outflow to the heart in 10 healthy adults. Afferent carotid baroreceptor activity was modified with 30-mmHg neck suction or pressure applied during held expiration, and efferent sympathetic activity was measured with microelectrodes inserted percutaneously into peroneal nerve muscle fascicles. Sympathetic responses were conditioned importantly by directional changes of carotid transmural pressure: increased pressure (onset of neck suction or offset of neck pressure) inhibited (totally) sympathetic activity, and reduced pressure (offset of neck suction or onset of neck pressure) augmented sympathetic activity. Responses occurred after a latency of about 2 s and did not persist as long as changes of neck-chamber pressure. Cardiac intervals were prolonged by increased carotid transmural pressures and shortened by decreased carotid transmural pressures, but, in contrast to sympathetic responses, cardiac responses adapted only slightly during neck-chamber pressure changes. Our results suggest that in the human a common baroreceptor input is processed differently in central vagal and sympathetic networks. Muscle sympathetic responses to changing levels of afferent baroreceptor traffic are profound but transitory. They appear to be conditioned more by changes of arterial pressure than by its absolute levels. PMID- 6278946 TI - Chloride self-exchange in toad skeletal muscle in vivo and in vitro. AB - The exchange of cellular Cl with 36Cl has been measured in saline-perfused hindlimb muscles of the pithed toad and compared with cellular Cl exchange in isolated muscles incubated in the vitro either in toad Ringer solution or in toad plasma. In the perfused hindlimb, the rate of 36Cl efflux from muscle cells [17.0 +/- 0.9 pmol Cl.(cm2 plasma membrane.s)-1] was only 40% as fast as that of the 36Cl influx. The discrepancy between Cl influx and efflux was accompanied by a cellular accumulation of Cl against the electrochemical gradient for this anion. Concurrently, the cells took up Na in amounts at least equal to the accumulated Cl. During this accumulation of Na and Cl, the mean resting potential remained constant at a value of -89.2 +/- 1.9 (SE) mV. Na and Cl were taken up by the muscle cells of the perfused hindlimb without a concomitant decrease in cellular K content; i.e., without evidence of inhibition of the Na-K pump. The rate constant for cellular 36Cl efflux from isolated toad muscles preincubated for 3 h in vitro in toad Ringer solution was about five times faster than that of muscles in the perfused hindlimb and similar in magnitude to published values for Cl fluxes in frog muscle. Cellular Cl efflux from muscles briefly preincubated in vitro for 15 min instead of 3 h was significantly slower than after prolonged preincubation. In vitro incubation of isolated toad muscles in toad plasma slowed the cellular 36Cl efflux to values approaching those measured in the perfused hindlimb, without comparably depressing the 36Cl influx. It is suggested that the uptake of NaCl by the cells of perfused hindlimb muscle may proceed by an electroneutral inward cotransport of Na and Cl on the same carrier. PMID- 6278947 TI - Catecholamine sensitivity in brown fat cells from cold-acclimated hamsters and rats. AB - Brown fat cells, freshly isolated from cold-acclimated hamsters and rats, did not respond to norepinephrine addition with the characteristic increase in oxygen consumption (heat production) seen in cells from control animals. However, incubation of these cells for 1 h in a Krebs-Ringer bicarbonate buffer, in the presence of 10 mM pyruvate, fully restored norepinephrine responsiveness. Cells treated in this way from cold-acclimated hamsters (a hibernator) increased the rate of oxygen consumption after maximal norepinephrine stimulation as much as cells from control hamsters; also norepinephrine-stimulated fatty acid release was unaltered, indicating that brown fat cells may partly be responsible for the increase in serum fatty acid level seen during arousal from hibernation. Similarly, preincubated cells from cold-acclimated rats (a nonhibernator) increased oxygen consumption and fatty acid release as much as cells from control rats; this suggests that also in cold-acclimated rats brown fat may supply the circulation with fatty acids during cold stress. Cells from cold-acclimated animals were, however, about 10 times less sensitive to norepinephrine than cells from control animals; this desensitization may be the result of a stimulated phosphodiesterase. PMID- 6278948 TI - Renal parathyroid hormone receptors in the chick: downregulation in secondary hyperparathyroid animal models. AB - We characterized the binding of 125I-[Nle8, Nle18, Tyr34]parathyroid hormone-(1 34) amide [125I-nlPTH-(1-34)] to renal plasma membranes prepared from chicks to determine the effects of secondary hyperparathyroid states on renal PTH receptors. This radioligand exhibited specific binding to membranes with high affinity (Kd, 2-3 X 10(-9) M). Agonists or competitive antagonists of PTH were effective in competing for binding sites labeled with 125I-nlPTH-(1-34), whereas an inactive fragment of PTH, salmon calcitonin, and bovine growth hormone did not compete with the radioligand for renal PTH receptors. Newly hatched chicks raised on control diet with adequate vitamin D and calcium or diets deficient in either vitamin D or calcium were used to study the regulation of renal PTH receptors in experimental models of secondary hyperparathyroidism. We found that both experimental diets resulted in marked hypocalcemia and progressive loss of renal cyclic AMP responsiveness to PTH in vitro. Associated with this refractoriness to the hormone was a marked reduction in PTH receptors in membranes from both vitamin D-deficient and calcium-deficient chick kidney. No change in the affinity of the PTH receptors was found. Vitamin D3, in a single dose of 250 micrograms, partially restored serum calcium of vitamin D-deficient birds toward normal by 72 h and also partly restored renal cyclic AMP responsiveness to PTH and the PTH receptor number toward control values. We conclude that renal refractoriness to PTH observed in experimentally hyperparathyroid animals models is due to a marked loss of plasma membrane receptor sites for PTH without an apparent change in the affinity of the receptors for the hormone. PMID- 6278949 TI - Renal Na-K-ATPase: its role in tubular sodium and potassium transport. AB - Na-K-ATPase, the enzymatic equivalent of the sodium:potassium pump, is found in large amounts in the kidney, and this organ has figured prominently both as a source for the purification of the enzyme and as a target for the study of its properties. Located on the basolateral aspect of tubule cells, renal Na-K-ATPase plays a key role in the active translocation of Na and K across this membrane as well as in the "secondary active" transport of a number of other solutes. The activity of renal Na-K-ATPase varies in parallel with sustained changes in Na or K transport, indicating the participation of this enzyme in the chronic adaptation of the kidney to altered Na reabsorption or K secretory load. Because of its slow turnover, however, the role of Na-K-ATPase in the modulation of acute changes in cation transport is unclear. Several hormones and vanadate influence renal Na-K-ATPase activity, and their importance as potential physiologic regulators of this enzyme is examined. Most of the information on the renal enzyme has been obtained from studies using homogenates or subcellular fractions thereof, but more recently the development of tubule microdissection and microanalytic methods has made possible the study of Na-K-ATPase in single nephron segments. This approach has opened new possibilities for evaluating the role of this enzyme in kidney function by facilitating correlation of enzyme activity with transport events in the same structure and by enabling us to focus the study of Na-K-ATPase on discrete anatomic subdivisions of the functionally heterogeneous nephron. PMID- 6278950 TI - Membrane fluidity and enzyme activities in brush border and basolateral membranes of the dog kidney. AB - The physical state of membrane lipids and relationships with the activity of Na+ K+-ATPase and alkaline phosphatase were studied in basolateral and brush border membranes of the dog kidney. Fluorescence polarization and electron spin resonance experiments demonstrate that basolateral membranes are much more fluid than brush border membranes. This can be accounted for by a difference in fluidity of the lipid part of the membranes. Broad (43-17 degrees C) thermotropic transitions are observed in liposomes made from total lipid extracts of brush border and basolateral membranes. Fluorescence data strongly suggest that thermotropic transitions also occur in intact membranes and that a change in membrane physical state may take place around the physiological temperature. A nonlinear Arrhenius plot for the Na+-K+-ATPase activity in basolateral membranes (breakpoint 21 degrees C) provides additional support for the existence of a lipid liquid leads to gel transition in antiluminal plasma membranes. A break in the Arrhenius plot of alkaline phosphatase activity is also observed but at a temperature significantly higher (26 degrees C) than that of the end of the thermotropic transition. "Room temperature" appears as a critical zone for lipid physical state and activities of both enzymes. PMID- 6278951 TI - Energetics of tubular sodium reabsorption sensitive to ethacrynic acid and ouabain. AB - Ouabain reduces renal oxygen consumption more extensively than ethacrynic acid despite similar natriuretic effects. Therefore, ethacrynic acid, which does not inhibit Na-K-ATPase, might stimulate energy metabolism unrelated to net sodium reabsorption. Experiments were performed on anesthetized dogs that had received isotonic saline intravenously corresponding to 10% of body wt and acetazolamide (100 mg.kg-1 i.v.). Subsequent infusion of ouabain in nine dogs (120 nmol.kg-1 intrarenally) reduced sodium reabsorption and oxygen consumption in parallel, giving a delta Na/delta O2 ratio of 18.0 +/- 1.1. With ethacrynic acid (3 mg.kg-1 i.v.) in six other dogs the delta Na/delta O2 ratio averaged 24.5 +/- 1.4. In a third group of five dogs, ouabain administered after ethacrynic acid reduced sodium reabsorption and oxygen consumption to the same levels as when ouabain was given alone. Thus, the high oxygen consumption remaining after ethacrynic acid can be inhibited by ouabain. We propose that ethacrynic acid generates a futile cycling of sodium by Na-K-ATPase across the basolateral cell membrane that is not apparent as net sodium reabsorption and is stopped by ouabain.U PMID- 6278952 TI - alpha 2-Adrenergic receptor regulation of ion transport in rabbit ileum. AB - Catecholamines are known to decrease short-circuit current (Isc), stimulate NaCl absorption, and inhibit HCO3 secretion in rabbit ileal mucosa in vitro. These effects appear to be mediated by alpha-adrenergic receptors because they are partially blocked by phentolamine and not by propranolol. To further characterize this receptor system, we determined the interactions of epinephrine (Epi) with alpha-subtype-selective antagonists. Prazosin (PZ), a specific alpha 1 antagonist, did not alter the Epi dose-response curve at concentrations up to 10( 5) M. Yohimbine (YO), a specific alpha 2-antagonist, completely inhibited the Epi effect on Isc. At 10(-5) M, YO increased by 70-fold the concentration of Epi required to produce a half-maximal effect (ED50; from 1.4 X 10(-7) M to 10(-5) M). YO and PZ by themselves had no significant effect on Isc in concentrations up to 10(-5) M. Clonidine, a specific alpha 2-agonist, decreased Isc with an ED50 similar to that of Epi; its effect was blocked by YO but not by PZ. Two alpha 1 selective agonists, methoxamine and phenylephrine, only caused a decrease in Isc in doses greater than 10(-5) M. This effect was reversed by YO but not by PZ. The effects of YO and PZ on Epi-modified Cl fluxes were also determined. YO completely aborted the effects of Epi on net Cl flux. No significant effects were seen with PZ. We conclude that the effects of Epi on ileal ion transport are mediated by a specific alpha 2-adrenergic receptor present in ileal mucosa and that no physiologic alpha 1-receptor function can be demonstrated. PMID- 6278953 TI - Mucosal gastrin receptor. VII. Up- and downregulation. AB - A series of experiments were performed to examine the long-term upregulation of the gastrin receptor and to explore the possibility of a short-term down regulation with regard to the time course of the trophic action of gastrin. Vagotomy or fasting was used to examine upregulation. Elevated serum gastrin levels observed postvagotomy are accompanied by an increase in gastrin binding capacity. Conversely, fasting caused a decline in serum gastrin and receptor levels; refeeding restored both values to normal levels. Cycloheximide was used to study the role of protein synthesis in the upregulation of gastrin receptors in vagotomized and refed rats, and it prevented the rise of gastrin binding capacity to either normal or above-normal values. This indicates that protein synthesis may play an integral part of receptor upregulation. Following the injection of gastrin-17, there was a short-term induction of downregulation. These results coupled with the maximal reduction of gastrin receptor 3 h after cycloheximide injection indicate that the gastrin receptor has a short half-life. Following the gastrin receptor interaction, the cellular response occurs at discrete periods and manifests itself as both down- and upregulation. PMID- 6278954 TI - Effects of diltiazem on smooth muscles and neuromuscular junction in the mesenteric artery. AB - Effects of diltiazem on membrane properties, neuromuscular transmissions, and mechanical responses were investigated in intact and skinned muscles of the guinea pig mesenteric artery. Diltiazem (greater than 10(-6) M) depolarized the membrane, increased the membrane resistance, and suppressed the spike evoked by either electrical depolarization or summation of excitatory junction potentials (EJPs). This drug also suppressed the facilitation process of amplitudes of EJPs produced by repetitive perivascular nerve stimulations. These suppressions of EJPs were not caused by a reduced number of active nerves contributing to the generation of EJP but but rather to a reduction in the release of chemical transmitters. Norepinephrine (NE)-induced and K-induced contractions were suppressed by diltiazem noncompetitively, but the contraction evoked in Na deficient solution was not suppressed, i.e., diltiazem is not a nonselective inhibitor of the Ca influx. In the saponin-treated skinned muscles diltiazem did not suppress the release from or the accumulation into the Ca store site, nor did it suppress activation of the Ca receptor in contractile proteins. These results indicate that diltiazem acts on the surface membrane and nerve terminal as a Ca antagonist or Ca-channel blocker. PMID- 6278955 TI - Myocardial responses to alpha-adrenoceptor stimulation with methoxamine hydrochloride in lambs. AB - This study was undertaken to evaluate the effects of methoxamine on left ventricular performance, coronary blood flow (CBF), and myocardial oxygen consumption (MVO2) in lambs. Measurement of maximum rate of rise of left ventricular pressure (LV dP/dtmax), left ventricular end-diastolic pressure (LVEDP), CBF, and MVO2 were made in nine lambs using a hemodynamically controlled preparation. The lambs were given atropine sulfate (1 mg), tetraethylammonium chloride (100 mg), and practolol (4 mg/kg) to provide parasympathetic, ganglionic, and beta-adrenergic blockades. Aortic pressure and blood flow and heart rate were held constant in each lamb. Dose-related increases in LV dP/dtmax and decreases in LVEDP were observed after progressively larger doses of methoxamine ranging from 0.4 to 6.0 mg/kg were given. These positive inotropic responses were eliminated by giving phentolamine (2 mg/kg). CBF, myocardial O2 extraction, and MVO2 did not change significantly. A positive inotropic action of methoxamine was also demonstrated in five additional animals by obtaining ventricular function curves. Initially the mean stroke volume at LVEDP 10 cmH2O (SV10) was 4.11 +/- 0.16 (SE) ml. This value increased to 5.09 +/- 0.28 ml after methoxamine (P less than 0.01). After phentolamine, SV 10 fell to 4.37 +/- 0.08 ml (P less than 0.05). These observations support the hypothesis that alpha adrenergic receptors are present and mediate a substantial positive inotropic action on neonatal lamb myocardium. PMID- 6278956 TI - Effects of fatty acid intermediates on Na+-K+-ATPase activity of cardiac sarcolemma. AB - The effect of amphiphilic lipid intermediates on the Na+-stimulatable activity of the Na+-K+-ATPase of sarcolemma from adult canine cardiac myocytes was studied. Sarcolemma (mean Na+-stimulatable ATPase activity of 73 mumol.mg sarcolemmal protein-1.h-1) was preincubated (37 degrees C for 10 min at pH 7.2) or rapidly mixed at 0 degrees C with amphiphilic lipid intermediates prior to dilution and assay of enzyme activity. Their effects were dependent on temperature, initial concentration, and the ratio of bound amphiphile to sarcolemmal protein. In particular, pretreatment of freshly prepared sarcolemma at 0 degrees C with arachidonyl CoA (up to 0.25 mM) caused 110% stimulation above control activity; palmitoyl CoA or palmitoyl carnitine under the same conditions caused no significant effect. Despite strong binding to the sarcolemmal vesicles, palmitoyl carnitine (up to 0.4 mM or 5 mumol/mg protein) and palmitoyl CoA (0.1 mM or 1.0 mumol of membrane-bound palmitoyl CoA/mg protein) were ineffective even with preincubation. Palmitoyl CoA was inhibitory above this level. Preincubation (22 degrees C for 10 min) with lysophosphatidylcholine only produced inhibition (40% at 0.75 mM). Thus fatty acyl thioesters of CoA and lysophosphatidyl choline but not palmitoyl carnitine perturb sarcolemmal Na+-K+-ATPase activity. PMID- 6278957 TI - Entrainment of the circadian rhythm from the eye of Aplysia: role of serotonin. AB - The finding that serotonin (5-HT) treatments as short as 1.5 h in duration produce phase shifts in a circadian rhythm from the isolated eye of Aplysia suggested that release of 5-HT was part of an ocular entrainment pathway. Since light cycles entrain this rhythm, we compared phase shifting by 5-HT and by light. The similarity in the shapes of the phase-response curves for 5-HT and light pulses indicates that 5-HT treatments are capable of entraining the rhythm. Also, "skeleton" 5-HT treatments phase shift as well as continuous 5-HT treatments. However, 5-HT does not appear to mediate the phase shifts produced by light, since 1) treatments that should block transmitter release do not change the phase shifts produced by light pulses; 2) the response curves of 5-HT and light pulses are displaced by 12 h relative to one another on the phase axis of the response curve; and 3) light-induced phase shifts are apparent almost immediately, whereas 5-HT-induced phase shifts become evident only about 24 h after 5-HT treatment. The eye appears to contain two independent entrainment pathways, one for light and one utilizing 5-HT. PMID- 6278958 TI - Developmental aspects of pituitary and adrenal responses to arterial hypotension in neonatal, weanling, and adult sheep. AB - We have studied neuroendocrine and heart rate responses to arterial hypotension in unanesthetized, chronically cannulated neonatal (less than 10 days old), weanling (3- to 4-wk-old), and adult sheep. Nitroprusside-induced arterial hypotension in the absence of hypovolemia promptly increased plasma ACTH, vasopressin (AVP), and cortisol levels in all three groups. The integrated adenohypophyseal and neurohypophyseal responses to a standard hypotensive stimulus were similar at the three ages, suggesting that the functional development of the systems subserving these responses is complete by the immediate postnatal period. The integrated cortisol/ACTH ratio was greatest in the youngest animals, suggesting that enhanced adrenal responsiveness to ACTH exists in neonatal as well as in late-gestation fetal lambs. The reflex tachycardia accompanying the arterial hypotension in the weanling and adult sheep was absent in the neonates. PMID- 6278959 TI - Reduced norepinephrine turnover in brown adipose tissue of ob/ob mice. AB - Obese (ob/ob) mice have a lower thermogenic capacity than lean mice. The possible role of brown adipose tissue (BAT) in this defect was investigated. Lean and obese mice were exposed to 33 (thermoneutral), 25, or 14 degrees C for up to 3 wk. BAT cytochrome oxidase activity and numbers of Na+-K+-ATPase enzyme units, enzymes involved in thermogenesis, were similar at 33 or 25 degrees C. Chronic exposure to 14 degrees C increased these enzymes 34 and 62%, respectively, in lean mice and nearly 150% in obese mice. Sympathetic nervous system activity, which stimulates thermogenesis in BAT, was evaluated by measuring norepinephrine (NE) turnover. At 25 degrees C, NE turnover rate in BAT of obese mice was only 40% as rapid as in BAT of lean mice. Chronic exposure to 33 degrees C depressed NE turnover in BAT of lean mice, but not in obese mice, whereas exposure to 14 degrees C accelerated NE turnover in both lean and obese mice. Lower sympathetic nervous system activity in BAT of obese mice at 25 degrees C is likely a major factor in their reduced nonshivering thermogenesis and resultant high efficiency of energy storage. PMID- 6278960 TI - Heterogeneity of sodium-dependent D-glucose transport sites along the proximal tubule: evidence from vesicle studies. AB - The glucose transport properties of brush border membrane vesicles from the outer cortex (early proximal tubule) and outer medulla (late proximal tubule) of rabbit kidney were studied. In the outer cortical preparation the behavior of the sodium dependent component of D-glucose flux indicated the presence of a low-affinity transport system with Km congruent to 6 mM and Vmax congruent to 10 nmol.min-1.mg protein-1 as measured under zero trans conditions at 40 mM NaCl and 17 degrees C. By contrast, in the outer medullary preparation this component of flux behaved as a high-affinity system with Km congruent to 0.35 mM and Vmax congruent to 4 nmol.min-1.mg protein-1. Differences in transport specificity between the two preparations were also indicated and glucose uptake by the outer cortical vesicles was significantly more sensitive to inhibition by phlorizin. These results suggest the existence of two distinct sodium-dependent D-glucose transport systems in the renal proximal tubule brush border membrane. The kinetic studies presented here were done under zero trans sodium and glucose conditions. The rationale and methodology for carrying out these measurements reliably are discussed in detail. PMID- 6278961 TI - Escherichia coli heat-stable toxin: its effect on motility of the small intestine. AB - Escherichia coli heat-stable enterotoxin is a low-molecular-weight substance that has been shown to induce the active secretion of fluid and electrolytes in the small intestine. In this study, we have characterized the effects of purified E. coli heat-stable toxin (ST, strain 18D, serotype 042:K86:H37) on the motility of rabbit small intestine by using myoelectric recording techniques. Substances, such as cholera toxin, that activate the adenylate cyclase-cAMP system induced predominantly migrating action-potential complex activity. E. coli ST, a toxin that activates the guanylate cyclase-cGMP system, was infused into isolated in vivo ileal loops of New Zealand White rabbits. Inactivated toxin was also studied by exposing the ST to 1 mM dithiothreitol for 90 min. Active E. coli ST induced only repetitive bursts of action potentials. When the toxin was inactivated with dithiothreitol, no alteration in myoelectric activity was observed. We speculate that repetitive bursts of action-potential activity may represent a virulent factor of the bacterium, altering motor activity to slow transit and allowing for bacterial proliferation and invasion. PMID- 6278962 TI - Receptors on smooth muscle cells: characterization by contraction and specific antagonists. AB - Smooth muscle cells were isolated from the stomach of the guinea pig, and the kinetics, stoichiometry, and specificity of contraction in response to the C terminal octapeptides of cholecystokinin (CCK-OP), gastrin-17, and acetylcholine were examined. All three agonists elicited dose-dependent peak contraction that did not depend on the presence of extra-cellular calcium. The potencies of CCK-OP and gastrin-17 were equal (D50, 10(-11) M) and 10 times greater than the potency of acetylcholine (D50, 10(-10) M). A combination of low doses of acetylcholine and CCK-OP was synergistic; however, its effect did not exceed the maximal responses to either agonists alone or to high extracellular concentrations of calcium. The specificity of the receptors was established by the use of atropine and the two CCK-receptor antagonists dibutyryl cGMP and proglumide. The span of the dose-response curves was wide, suggesting the existence of receptor heterogeneity. It is concluded that gastric smooth muscle cells of the guinea pig possess distinct, high-affinity receptors for CCK-gastrin and acetylcholine; the receptors mediate contraction that is not immediately dependent on the presence of extracellular calcium. PMID- 6278963 TI - Prevention of phenylbutazone ulcer in the rat by glucose: role of a glycoprivic receptor system. AB - The mechanism by which the extragastric administration of phenylbutazone (PBZ) and other nonsteroid anti-inflammatory drugs induce a vagally mediated gastric hypercontractile response and ulceration is unknown. Glucose administration has been reported to protect the rat and the monkey from the ulcerogenic action of aspirin. It was postulated that by uncoupling oxidative phosphorylation PBZ might stimulate an energy-sensitive receptor system that results in the gastric contractile response. Glucose feeding or intravenous infusion was seen to prevent or abolish the gastric contractile response to the intraileal injection of 22.5 mg of PBZ in cervical-sectioned rats with miniature gastric balloons. Glucose treatment also significantly reduced ulceration and hemorrhage. Fructose infusion was somewhat less effective. Mannitol infusion had no effect on the PBZ-induced gastric contractile activity, and all animals developed severe ulceration and hemorrhage. Blood glucose levels fell significantly in starved rats treated with PBZ. Glucose feeding and glucose or fructose infusion prevented or reduced this effect. It is suggested that by alleviating the "glucoprivic state" induced by PBZ glucose abolishes the gastric contractile response and prevents ulceration and hemorrhage. PMID- 6278964 TI - Postnatal regulation of canine oxygen delivery: control of erythrocyte 2,3-DPG levels. AB - The oxygen affinity of canine blood changes markedly following birth. These changes are correlated with alterations in the intracellular concentration of 2,3 diphosphoglycerate (2,3-DPG). We have examined the control of erythrocyte glycolysis by measurements of intracellular enzymes and intermediates, and we have identified the component responsible for regulation of 2,3-DPG concentration and hence blood oxygen affinity during canine postnatal development. The concentration of 2,3-DPG could be regulated entirely by the enzymes of the Rapoport-Luebering shunt. We have not detected any alterations in the levels or intracellular activity of 2,3-DPG mutase or 2,3-DPG phosphatase during development; therefore postnatal changes of 2,3-DPG must be a result of changes in the intracellular concentrations of 1,3-diphosphoglycerate (1,3-DPG) that are controlled by other reactions in the glycolytic pathway. Neither low intracellular concentrations of glucose, the glycolytic substrate, nor an inherently low glycolytic rate can account for the low 2,3-DPG levels at birth. 1,3-DPG concentrations and hence 2,3-DPG concentrations are controlled by the activity of pyruvate kinase, which acts as a glycolytic sink reaction. The intracellular activity of pyruvate kinase decreases during the first 50-60 days of age and causes the accumulation of 2,3-DPG. There is a subsequent change in the in vivo kinetic properties of the enzyme, giving increased intracellular activity and resulting in the slow decline of 2,3-DPG concentrations toward normal adult values. PMID- 6278965 TI - Evidence for a neurotransmitter role for epinephrine derived from the adrenal medulla. AB - We examined the neurohumoral mechanisms underlying the hindlimb vasodilator response produced by electrical stimulation of the anteroventral region of the third ventricle (AV3V). Hindlimb blood flow velocity was recorded using a pulsed Doppler flow probe. The vasodilator response to AV3V stimulation was greater than that obtained after inhibition of neurogenic vasoconstrictor tone with sympathectomy and was, therefore, in part an active process. The hindlimb vasodilator response was not affected by cholinergic or histaminergic receptor blockade but was reduced by bilateral adrenalectomy (ADX) or adrenal demedullation (ADM) and was further reduced by beta-adrenergic receptor blockade with propranolol in ADX rats. In ADX or ADM rats the vasodilator response was attenuated by repeated AV3V stimulations and restored by epinephrine infusion. Moreover, restoration of the response after epinephrine infusion was completely blocked by the neuronal uptake blocker desmethylimipramine. As was observed with vasodilation, constrictor responses to AV3V stimulation in the renal and mesenteric vascular beds were also attenuated by adrenal demedullation and were restored by epinephrine infusion. These data suggest that circulating epinephrine, originating in the adrenal medulla, is taken up by sympathetic nerve terminals innervating blood vessels. The catecholamine is then released from these nerves and acts like a classical neurotransmitter, producing vasodilation in skeletal muscle and vasoconstriction in splanchnic and renal vascular beds. PMID- 6278966 TI - alpha- and beta-Adrenergic effects of epinephrine on ventricular pacemakers in dogs. AB - We studied the effects of epinephrine on idioventricular rhythm in 15 adult dogs with chronic complete heart block induced by the injection of formalin into the His bundle. Atropine (0.1 mg/kg) was given intravenously to attenuate any potential vagal effects, and epinephrine was infused in graded doses of 0.01-10.0 micrograms.kg-1.min-1. Two different responses were seen. In 12 dogs there was a concentration-dependent increase in ventricular rate following epinephrine infusion. These animals then were given the beta-blocker propranolol (0.5 mg/kg iv), and the epinephrine infusions were repeated. In this situation epinephrine concentrations less than 0.1 micrograms.kg-1.min-1 induced a decrease in ventricular rate with no associated change in arterial pressure. In four additional dogs this decrease in ventricular rate was prevented by alpha-blockade with phentolamine. In three dogs epinephrine less than 0.1 micrograms.kg-1.min-1 induced a decrease in ventricular rate without an associated change in arterial blood pressure. This decrease in rate was abolished by the alpha-blocker phentolamine. It therefore appears that an alpha-adrenergic effect on ventricular automaticity can occur in the intact animals. When this does not occur initially, it can be unmasked by propranolol and results in a slowing of ventricular rate unrelated to changes in blood pressure. PMID- 6278967 TI - Histocompatibility antigens in pregnancy, abortions, infertility, preeclampsia, and trophoblast neoplasms. AB - The genetic makeup of an individual may determine the capacity of the individual to combat successfully cancer and other disease states, eg abnormal pregnancies, abortions, infertility, and preeclampsia. These factors appear to be based upon the individual's immunogenetics, as histocompatibility-restricted T-cell cytotoxicity may be deficient in the observed instances of increased histocompatibility between husband and wife found in trophoblast tumor patients and in other instances of abnormal pregnancies. PMID- 6278968 TI - The IgG and transferrin receptors of the human syncytiotrophoblast microvillus plasma membrane. AB - Fc gamma receptor activity has been demonstrated using a radioisotopic assay on human syncytiotrophoblast microvillus plasma membrane (StMPM) vesicle preparations isolated from normal full-term placentas. Prior chaotropic extraction to remove extrinsic membrane protein from StMPM preparations was shown to increase radiolabeled IgG uptake by up to 30%. Polyacrylamide disc gel electrophoresis (PAGE) has been used to indicate that the StMPM receptor for transferrin solubilizes in 1% sodium deoxycholate and retains its binding activity for transferrin. Quantitative data for both Fc gamma and transferrin receptors obtained following study of isolated StMPM preparations are summarized. Current concepts for the potential role of trophoblast Fc gamma and transferrin receptors in the mechanisms of transplacental passage of IgG and iron, respectively, from mother to fetus are illustrated and discussed. PMID- 6278969 TI - Cell-mediated immunity to cytomegalovirus in pregnant women. AB - Employing the techniques of in vitro lymphocyte transformation (LTF) and complement fixation, cell-mediated immunity (CMI) and antibody to cytomegalovirus (CMV) were studied in pregnant and nonpregnant women. The LTF activity was determined by the whole blood microassay using four strains of CMV (AD-169 and its early antigen [EA], Davis, Veca, and Towne strains), and phytohemagglutinin (PHA). Lymphocyte transformation response to specific CMV antigens at 11-30 weeks of gestation and to nonspecific mitogen (PHA) in all pregnant and postpartum women were found to be significantly depressed compared with the nonpregnant women. The lower LTF responses to CMV antigen and PHA were found in specimens taken from pregnant women at 21-30 weeks of gestation. There were no significant differences in the mean complement-fixing (CF) antibody titers and the percentage of E-rosette-forming T lymphocytes between subjects in various stages of pregnancy. In addition, concanavalin A (Con A)-generated suppressor T cell activity was evaluated in pregnant and nonpregnant women. The suppressor effect of Con A-activated lymphocytes in the pregnant women was somewhat higher than in nonpregnant women. These observations suggest that CMV-specific suppression of cellular immunity may play an important role in reactivation of CMV in pregnancy. PMID- 6278970 TI - Human papillomavirus (HPV) in condylomatous lesions of cervix. AB - Ninety-seven cervical condylomata classified histologically as flat condyloma (planum), papillary condyloma (acuminatum), and endophytic condyloma were studied by transmission electron microscopy (TEM) and by immunoperoxidase technique (IPT) for the presence of human papillomavirus (HPV) particles and antigen, respectively. Both techniques localized HPV chiefly in nuclei of koilocytotic cells. HPV particles were found in 25% of the cases by TEM and HPV antigen was detected in 48% of the cases by IPT. All cases positive by TEM were also positive by IPT, thus confirming the specificity of the immunological staining. The viral antigen was detected in 56% of 68 flat condylomata, 35% of 26 papillary condylomata, and in none of 3 cases of endophytic condylomata. However, when histiotypes of virus-positive condylomata were controlled for the intraepithelial extent of koilcytotic cells, the prevalence of HPV correlated with the extent of koilocytosis rather than with the histiotype. The immunologic technique will be of value for the further characterization of cervical condylomata and of the relationship between HPV infection and cervical intraepithelial neoplasia. PMID- 6278971 TI - Prostatic mucinous adenocarcinoma. A Cowper gland carcinoma mimicker. PMID- 6278972 TI - Melanotic adenocarcinoma of the anorectum. AB - Epithelial cells of an otherwise typical anorectal mucinous adenocarcinoma contained melanin pigment. Ultrastructurally, three cell populations were present. The largest population contained membrane-bound compound melanosomes in cells having glandular epithelial characteristics including surface specializations, microvilli, and mucin. Additionally, occasional melanocytes were intermixed with the tumor cells and finally very rare cells having both mucin and isolated melanosomes were present. We believe that this phenomenon is better explained by phagocytosis or transfer of melanin from melanocytes to neoplastic cells, rather than by melanin synthesis in adenocarcinoma. PMID- 6278973 TI - Hemangiopericytoma of the breast. AB - The clinical and pathologic features of three hemangiopericytomas of the breast in women are described. The three tumors presented as painless, nontender nodules. The morphologic variation typical of hemangiopericytomas was present in each tumor. One case was studied by electron microscopy and was found to be composed of two types of pericytes. Both cell types were closely associated with vessels, partially or completely surrounded by basal lamina, and contained pinocytotic vesicles. There were only minor differences between the two forms of pericytes. Smooth muscle cells, fibroblasts, or transitional cell forms were not observed. The two patients on whom follow-up information was available have remained free of tumor 2.8 and 4 years after their surgery, indicating that not all vascular tumors of the breast are highly aggressive neoplasms. PMID- 6278974 TI - Recurrent malignant fibrous histiocytoma. A histologic analysis of 50 cases. AB - This clinicopathologic study deals with 84 patients who had recurrent malignant fibrous histiocytoma (MFH), and is primarily concerned with 50 cases in which specimens of both the initial and recurrent tumors were available for histopathologic examination. Quantitative and qualitative differences in the histologic patterns characterizing MFH, such as storiform and myxoid, between the initial and recurrent tumors, were clear-cut in nearly two-thirds of the cases. Conversion in histologic subtypes of MFH occurred in nine cases, including six cases which converted from myxoid to nonmyxoid pleomorphic. The superficially located tumors recurred somewhat more frequently, but in such cases, there was a significantly better prognosis than was seen with the deeply located tumors. Changes in the degree of mitotic rate, nuclear atypism, and cellularity from the initial to the recurrent tumor correlated better with the biological behavior of the tumor than did the degree in the initial tumor itself. The increased proportion of histiocyte-like tumor cells in the recurrent tumor was another morpholic feature which indicated a much poorer prognosis than did the increased proportion of fibroblast-like tumor cells. PMID- 6278975 TI - Mammary carcinoma with "choriocarcinomatous" features. PMID- 6278977 TI - Antigen-induced bronchial anaphylaxis in actively sensitized SD rats. Effects of immunization and provocation doses of antigen and of pretreatment with DSCG, theophylline, terbutaline and a new anti-allergic xanthine derivative, D4026. AB - Bronchial anaphylactic reactions, estimated as increase in intratracheal pressure, were precipitated by intravenous injections of antigen into actively sensitized SD rats. The degree of bronchial reactivity was found to depend on both the challenge dose and the immunization dose of antigen; therefore the course of the capacity to respond was recorded as a function of these variables. The degree of the bronchial anaphylactic response could be reduced by pretreatment with disodium cromoglycate, terbutaline or a new anti-allergic xanthine derivative, D 4026, in some groups of animals. The efficacy of each agent was found to depend on the dose of antigen used for sensitization and for provocation of the bronchial reaction rather than on the strength of the response. Taken together, the data suggest that more than one type of homocytotropic antibody mediates bronchial anaphylactic reactivity in the SD rat. PMID- 6278976 TI - Brain shrinkage and subdural effusion associated with ACTH administration. AB - Sequential computed tomographic (CT) studies of 11 patients (aged five months to seven years) with intractable epilepsy treated with synthetic ACTH-Z showed brain shrinkage in all cases. Brain shrinkage started to appear on daily ACTH injections for seven days, reached the maximum within four weeks of administration (14 injections every day and then 7 injections every other day), and almost returned to the original status in seven out of nine cases which were followed up for one to three months after the therapy. The subjects aged less than two years showed more remarkable brain shrinkage than did those aged more than five years. Furthermore, two other cases were complicated by subdural effusion after ACTH therapy. It is the authors' assumption that both of these phenomena are caused by the high concentration of corticosteroid through a change of the water and electrolyte contents in the brain. PMID- 6278978 TI - A simplified radioactive assay for the enzyme UDP-glucose pyrophosphorylase. PMID- 6278979 TI - A new discontinuous buffer system for the electrophoresis of cationic proteins at near-neutral pH. PMID- 6278981 TI - Analysis of leukotrienes by high-pressure liquid chromatography. PMID- 6278980 TI - Assay of short-chain acyl coenzyme A intermediates in tissue extracts by high pressure liquid chromatography. PMID- 6278982 TI - Effect of the incomplete separation of bound and free ligand on binding measurements. PMID- 6278983 TI - Rapid separation of high-mannose-type oligosaccharides by high-pressure liquid chromatography. PMID- 6278984 TI - Superoxide-dependent formation of hydroxyl radicals: detection of hydroxyl radicals by the hydroxylation of aromatic compounds. PMID- 6278985 TI - Sensitive detection of specific repair endonucleases: radial diffusion assay utilizing differential alkaline denaturation of supercoiled and nicked PM2 DNA in agarose gels. PMID- 6278986 TI - Correlation between growth rate and cytochemistry in Morris hepatomas. AB - The correlation between the cytochemistry (glycoprotein, glycogen, glucose-6 phosphatase, catalase, alkaline phosphatase) and the growth rate of the fast growing Morris hepatoma 3924A and the slow-growing Morris hepatoma 9618A was studied by utracytochemical techniques. By the chromic acid-phosphotungstic acid technique, acid glycoprotein is stained in glycocalyx, Golgi saccules and vesicles, and secretory granules of the tumor cells of both hepatomas. However, the hepatoma 3924A cells contain thicker glycocalyx and more numerous glycoprotein-rich granules than hepatoma 9618A cells. Abundant alpha and beta glycogen particles are found in hepatoma 3924A. Moderate glucose-6-phosphatase activity is observed in the cisternae of endoplasmic reticulum and nuclear envelope of hepatoma 9618A, but it is totally absent in hepatoma 3924A. High catalase activity is present in numerous peroxisomes of hepatoma 9618A. Hepatoma 3924A contains only a few catalase-positive microperoxisomes. Weak to moderate alkaline phosphatase is present in the plasma membrane and nuclear envelope of hepatoma 9618A cells, while hepatoma 3924A shows no activity of the enzyme. All the cytochemical parameters except glycoprotein show an inverse relationship with the growth rate of the hepatomas. The higher intracellular glycoprotein content of hepatoma 3924A may be related to differences in cell coat secretion (composition and activity) from the slower-growing hepatoma 9618A PMID- 6278987 TI - Nitrous oxide at analgesic concentrations--an opiate agonist: further evidence. PMID- 6278988 TI - Lithium and neuromuscular transmission. AB - The actions of lithium on end-plate depolarization and on indirect and direct twitch response of isolated guinea pig muscle were investigated. At clinical concentrations lithium did not cause depolarization, nor did it affect the depolarizing action of carbachol. Lithium also had no effect on the twitch response to nerve stimulation throughout its therapeutic serum concentration range and it did not alter the ED50 of pancuronium. When animals were chronically pretreated with lithium there appeared to be a slight reduction in dosage requirement for d-tubocurarine. At concentrations well above therapeutic serum levels, end-plate depolarization as well as indirect and direct twitch responses were depressed. It is concluded that, at therapeutic levels, there is minimal interaction of lithium with competitive neuromuscular blocking agents. PMID- 6278989 TI - Effect of bretylium on neuromuscular transmission. AB - The effect of bretylium on the response of an isolated muscle (guinea pig lumbrical) to direct and indirect stimulation was examined. An initial increase in twitch response to both indirect and direct stimulation was followed at higher concentrations of bretylium by block of the indirect response only. The ED50 for this latter effect was 106.9 +/- 9.25 microM. However, in the presence of subthreshold levels of d-tubocurarine the ED50 was markedly reduced to 1.31 +/- 1.11 microM, a concentration that could occur during clinical use of bretylium. Thus, the physician is cautioned to consider the possibility that neuromuscular block may be produced when bretylium is administered in close temporal proximity to neuromuscular blocking agents. PMID- 6278990 TI - Opiate receptors and their definition by antagonists. PMID- 6278992 TI - [Nature and dynamics of spontaneous decurarization with d-tubocurarine as a muscle relaxant]. PMID- 6278991 TI - Opiate receptor mediation of ketamine analgesia. AB - Previous workers have noted that analgesia produced by ketamine can be antagonized by the narcotic antagonist, naloxone. In order to elaborate further the apparent similarity between ketamine- and narcotic-induced analgesia, the authors examined the effects of ketamine in three standard test systems for the opiate receptor. In a radioligand binding assay using 3H-dihydromorphine, ketamine stereospecifically bound to opiate receptors in rat brain homogenate, (+) ketamine being 2-3 times more potent than the (-) enantiomer of ketamine. In a bioassay for the opiate receptor, using the longitudinal muscle-myenteric plexus of the guinea pig ileum, ketamine inhibited the twitch-like muscular contractions, as do narcotics. However, only the inhibitory effects of (+) ketamine, which in this system also was twice as potent as (-) ketamine, could be partially antagonized by naloxone, suggesting that this enantiomer is responsible for the opiate receptor-related effects of ketamine. In vivo, the authors found that ketamine displaces 3H-etorphine, a potent narcotic, from opiate receptors in regional areas of the mouse brain, especially in the thalamic region, but not in the cortex. The results suggest that a significant mechanism of ketamine-induced analgesia is mediated by opiate receptors. PMID- 6278993 TI - [Changes in the enterochromaffin and pituitary-adrenal systems in the immediate postoperative period in children with nephroblastomas]. PMID- 6278995 TI - Prevalence of bluetongue viral antibodies in Louisiana goats. PMID- 6278994 TI - Adenovirus enteritis in pigs. AB - Experimental transmissions were done with adenovirus strain 6618 in hysterectomy produced colostrum-deprived pigs. After an incubation period of 3 to 4 days, all inoculated animals had diarrhea. Histopathologically, many intranuclear inclusion bodies were present on short villi of the terminal parts of jejunum and ileum. With electron microscopy, the inclusion bodies were observed to contain numerous adenovirus particles. Immunoperoxidase-positive cells were seen on short villi of the terminal parts of jejunum and ileum. Adenovirus particles also were detected by negative staining of intestinal contents. In 1 pig (naturally occurring infection), adenovirus enteritis was studied by the aforementioned techniques. Similar intestinal lesions as described in the experimental pigs were observed. PMID- 6278996 TI - Isolation of transmissible Gastroenteritis virus, pseudorabies virus, and porcine enterovirus from pharyngeal swabs taken from market-weight-swine. AB - Pharyngeal swab samples were collected at a central Iowa abattoir from 6,010 market-weight slaughter hogs from September 1, 1979 to August 31, 1980. The swab samples were examined for cytopathic viruses by inoculation of monolayer cultures of continuous line of swine testicular cells. Of the 6,010 swab samples tested, transmissible gastroenteritis virus was isolated from 91 (1,51%), pseudorabies virus was isolated from 431 (7.17%), and porcine enterovirus was isolated from 21 (0.35%). Although all 3 viruses were identified throughout the year, transmissible gastroenteritis and pseudorabies viruses were found more frequently during the winter and early spring. In contrast, porcine enterovirus was detected more frequently during the spring and summer. PMID- 6278997 TI - Differentiating bronchioloalveolar carcinoma from adenocarcinoma. AB - The recognition of bronchioloalveolar carcinoma (BAC) as distinct from adenocarcinoma of the lung, is controversial. Using strict pathologic criteria, 43 consecutive patients with BAC were matched by year of diagnosis and compared with a similar number of patients with adenocarcinoma, and for contrast, with those with squamous and oat cell carcinoma of the lung. We demonstrated that BAC is not sex related, and is not as smoking related as the other neoplasms. Unlike epidermoid carcinoma, BAC does not show a predilection for those occupations requiring manual labor. Also, BAC is frequently distinguishable radiologically from the other three by being smaller and peripheral. A pleural tag and an air bronchogram in a mass are rather specific, and BAC is less likely to have large airway involvement and adenopathy. The percentage of patients who were free of tumor after 2 yr was greater in the BAC group than in the others, but the overall survival rate between the BAC group and the adenocarcinoma group was not. Based on inter-observer variability, there is some overlap pathologically between these 2 groups. However, when the overlap between the adenocarcinoma and the BAC groups is compared with that between the adenocarcinoma and the squamous cell carcinoma groups, the difference is not significant. We conclude that BAC should be considered a distinct clinical entity. PMID- 6278998 TI - Subsensitization of beta-adrenoceptors in airways and lymphocytes of healthy and asthmatic subjects. AB - Subsensitization of beta-adrenoceptors in airways and lymphocytes developing during 4 to 5 wk of orally administered terbutaline (a long-acting beta 2 selective bronchodilator) was compared in 10 healthy subjects and 11 subjects with mild asthma. The following results were obtained. Normal subjects developed a significant reduction in acute bronchodilator responsiveness to inhaled isoproterenol but not to subcutaneously administered terbutaline. The subjects with asthma failed to develop any alteration in responsiveness to either inhaled isoproterenol or subcutaneously administered terbutaline. None of 5 normal or 4 asthmatic subjects studied demonstrated any significant change in sensitivity to histamine-induced bronchospasm or any significant decrease in the acute protective effect of subcutaneously administered terbutaline against histamine induced bronchospasm. Marked and significant decreases occurred in the density of lymphocyte beta-receptor sites in both groups. Maximal isoproterenol-stimulated cyclic AMP responses in lymphocytes from normal subjects were reduced, but in asthmatics, in whom the baseline values were lower to begin with, a similar decrease was not observed. In most instances, the number of receptor sites returned to normal within 2 wk after cessation of terbutaline. A single dose of methylprednisolone caused a return to normal values within 16 h. We conclude that chronic therapy with an oral beta 2-adrenostimulant, although producing striking and significant down-regulation of beta-adrenergic receptors of peripheral lymphocytes, does not lead to physiologically detectable beta-adrenoceptor subsensitivity in the airways of asthmatics or to consistent subsensitivity in the airways of healthy persons. PMID- 6278999 TI - Talc granulomatosis: laboratory findings similar to sarcoidosis. AB - We studied 6 men who had some combination of dyspnea, abnormal chest radiographs, resting increased alveolar-arterial oxygen gradients, noncaseating granuloma by lung biopsies, increased serum angiotensin-converting enzyme concentrations, positive gallium lung scans, and increased lymphocyte counts by bronchoalveolar lavage. Our patients were suffering from talc granulomatosis secondary to the intravenous injection of suspended, crushed pentazocine tablets. In addition to the above findings, bronchoalveolar lavage revealed birefringent intracellular and extracellular particles consistent with talc. We conclude that talc granulomatosis can mimic the laboratory findings of pulmonary sarcoidosis, but the findings of birefringent particles in the bronchoalveolar lavage fluid by polarized microscopy is a simple laboratory method for confirming a clinical diagnosis. PMID- 6279000 TI - Effects of leukotriene E on pulmonary mechanics in the guinea pig. AB - The effects of intravenously infused 5(S)hydroxy-6(R)-S-cysteinyl-7,9, trans,11,14,-cis eicosatetraenoic acid (leukotriene E) (LTE), one of the leukotriene constituents of slow-reacting substance of anaphylaxis (SRS-A), on pulmonary resistance (RL) and dynamic compliance (Cdyn), breathing frequency, and mean systemic arterial pressure were determined in both anesthetized and unanesthetized guinea pigs. The LTE caused a dose-dependent increase of RL and decrease in Cdyn over the range of doses from 100 to 10,000 ng/kg with significance effects at the highest doses. The onset of effect after a significant dose occurred within 30 s and was maximal 1 to 3 min after infusion. The LTE elicits a significantly greater effect on RL for a given change in Cdyn than occurs with LTC or LTD indicating that LTE is a less selective peripheral airway agonist than LTC or LTD. The LTD infusion resembled LTC or LTD in evoking a systemic arterial hypotension that was preceded by a brief initial period of hypertension in unanesthetized animals. PMID- 6279001 TI - Pulmonary sarcoidosis: correlation of serum angiotensin-converting enzyme with blood and bronchoalveolar lymphocytes. AB - Blood and bronchoalveolar (BA) cell populations were compared with serum angiotensin-converting enzyme activity (SACE) in 22 smoking and nonsmoking patients with recently diagnosed or active sarcoidosis. There was no correlation between SACE and blood lymphocyte populations, but a significant correlation was found between the number of BA lymphocytes and SACE and the number of BA T cells and SACE. In no case was SACE elevated when the number of BA lymphocytes was normal, but in 10 patients with increased numbers of BA lymphocytes SACE was normal. In fact, SACE was increased in less than one-third (7/22) of the patients. These results suggest that both SACE and the number of lymphocytes in lavage fluid reflect the intensity of the lung's inflammatory response in sarcoidosis but that quantification of BA lymphocytes is a more sensitive measure than SACE of active pulmonary sarcoidosis. PMID- 6279002 TI - [Wiedemann-Beckwith syndrome with hepatoblastoma (author's transl)]. PMID- 6279003 TI - [Cytomegalovirus infection in 32 children (author's transl)]. AB - Thirty-two cases of cytomegalovirus infection are reviewed; 13 were congenital, 19 acquired. The clinical aspects of evolution and manifestation of the disease, agree with those described in other series. Virological diagnosis was based on isolating the virus from either urine or pharynx (or both) and culturing in pulmonary fibroblasts from human embryo. To obtain optimum results, fresh specimens must be used. Although several techniques are available for CMV antibody measurement, immunofluorescence is the technique of choice. Clinical manifestations differ according to age and immunological status of the patient. Late manifestations of congenital infection, monosymptomatic forms and acquired forms of the disease are at present of great interest. Importance of establishing diagnosis through virological isolation whenever a cytomegalovirus infection is suspected, is underlined. PMID- 6279004 TI - Ketoconazole for treatment of disseminated coccidioidomycosis. AB - Of 29 selected patients with disseminated coccidioidomycosis, 27 were treated for at least 6 months with ketoconazole, 200 to 600 mg/d. Two patients had progression of coccidioidal disease shortly after starting ketoconazole, and one developed meningitis. Seven of eight patients with synovitis had prompt improvement in symptoms, but four either had recurrent synovial thickening without recoverable Coccidioides immitis or could not remain free of symptoms off the drug. The response of osteomyelitis to ketoconazole was hard to assess; three of eight cases clearly improved and none progressed. Abscess or sinus formation clearly improved in eight of patients; five remained free of disease after the drug was discontinued. Skin lesions improved in six of nine; three lesions remain healed off the drug. Ketoconazole is absorbed readily after oral ingestion and has little toxicity. In the dosages used, it seems to suppress but not eradicate C. immitis. The drug may be able to stabilize the infection while cell-mediated immunity is restored. PMID- 6279005 TI - Ketoconazole for treatment of chronic pulmonary coccidioidomycosis. AB - Twenty-one patients with chronic pulmonary coccidioidomycosis were treated with ketoconazole. In 16 patients with chronic cavitary disease, nine improved, four showed no change, and the condition of three deteriorated; three of nine patients had culture conversion. Roentgenograms showed improvement in two patients, no change in 12, and deterioration in two. Serologic improvement was not noted. In five patients with persistent infiltrative disease, the response was more favorable. Radiographic improvement, culture conversion, and serologic improvement were seen in most patients. Ketoconazole-induced side effects were mild and of short duration, ending in the first days of therapy. Ketoconazole seems to be of significant value in infiltrative pulmonary disease, but seems unable, at the doses used, to change finding in the sputum or to change radiographic findings in chronic cavitary disease. Further long-term observation is necessary to evaluate fully the role of this drug in chronic pulmonary coccidioidomycosis. PMID- 6279006 TI - Peripheral neuropathy and starvation after gastric partitioning for morbid obesity. AB - Three months after gastric partitioning for morbid obesity, two patients developed an unusual and severe form of polyneuropathy that affected their sense of position maximally. This disorder produced severe ataxia of the upper extremities and trunk, and pseudochorea. One patient died and the autopsy showed an extensive demyelinating polyneuropathy. Neuronal cell bodies in the anterior horns and dorsal root ganglia showed extensive accumulations of lipofuscin and Schwann cells showed extensive accumulations of lipid. This neuronal and Schwann cell lipidosis appears to result from starvation of the obese and has never been reported in other forms of human starvation or nutritional deficiency. PMID- 6279007 TI - [Inappropriate elevation of plasma thyroid stimulating hormone]. PMID- 6279008 TI - [Application of hormone determination to the diagnosis of pituitary corticotropic adenomas]. PMID- 6279009 TI - [Neuroradiological surveillance of secretory pituitary adenoma]. PMID- 6279010 TI - [Hormonal surveillance of corticotropic adenoma]. PMID- 6279011 TI - [Bilateral parotid gland tumors. Report on 3 cases (author's transl)]. AB - Three cases of rarely observed bilateral parotid gland tumors are reported. In two cases the tumors were of different histological types (mixed tumor on one side and an adenocarcinoma on the other side) while in the third case they were both of the mixed variety. No clear explanation for the possible bilateral nature of these tumors was found, diagnosis being made either during the initial examination or after an interval of several years. PMID- 6279012 TI - Radiographic and scintigraphic appearance of bone metastasizing Wilms'tumour: problems in confirming diagnosis. PMID- 6279013 TI - Mechanisms of antibiotic-induced nephrotoxicity. PMID- 6279014 TI - The activity of five cephalosporins against Bacteroides fragilis. PMID- 6279015 TI - Racial differences in intact erythrocyte ion transport. AB - Epidemiologic studies indicate a much higher incidence of hypertension in blacks than in whites, although no clear biochemical correlates to account for such overt racial differences have been identified. In recent years, considerable evidence has linked perturbations in ion transport to the risk of developing hypertension. Potassium (K+) transport and the ouabain-sensitive component of K+ transport in the erythrocyte were measured in 54 healthy, age and sex matched black and white subjects. Blacks have a significantly (p less than 0.001) lower capacity for K+ transport (0.190 +/- 0.03 mumoles K+ per hr per 10(9) red blood cells [RBC] than whites (0.230 +/- 0.03 mumoles K+ per hr 10(9) RBC) with a significantly (p less than 0.001) higher percentage of K+ transport dependent upon ouabain-sensitive mechanisms (blacks 85.26 +/- 4.14 percent versus whites 76.69 +/- 6.67 percent). These data clearly define overt racial differences in K+ transport which suggest that blacks have a more limited capacity to exchange intracellular sodium for extracellular K+, and a higher percentage of that exchange is dependent upon ouabain-sensitive mechanisms. These findings need be kept in mind were clinical studies of ion transport are being assessed and may be germane to the increased prevalence in blacks for the development of hypertension. PMID- 6279016 TI - Characterization of an automated radioimmunoassay for T4, T3, T3 U, and FTI. AB - The performance characteristics of assays is reported for thyroxine (T4), triiodothyronine (T3), and T3-uptake (T3U) using the GAMMAFLOTM Automated Assay System. A comparison of calculated free thyroxine index (FTI) values is also presented. This automated radioimmunoassay (RIA) system utilizes a combination of continuous-flow methodology and chromatographic separation techniques. The T4 assay studied had a standard curve range of 1.5 to 24.0 microgram per dl. The intra- and inter-assay precisions were 4.3 and 5.3 percent CV, respectively, for a T4 concentration of 10.0 microgram per dl. The T3 assay had a standard curve range of 50 to 1000 ng per dl, the corresponding precisions were 7.3 and 7.1 percent CV, respectively, for a concentration of 213 ng per dl. The automated serum T4 and T3 results correlated (r = 0.966 and 0.864) with a manual radioimmunoassay procedure. Intra-assay and inter-assay precisions for a mid range normal 30.1 percent T3U value were 6.2 percent and 4.9 percent CV, respectively. Reference range comparison of FTI by both automated and manual results correlated for 47 out of 51 (95 percent) patients compared. It is concluded that this automated system appears to offer a viable alternative to T4, T3, and T3U manual RIA techniques in terms of operational simplicity, analytical performance, and sample through-put flexibility. PMID- 6279017 TI - Preliminary use of linear discriminant function to predict success or failure of kidney transplant. AB - Adenylate kinase (AK) activity appears in serum or in urine as a result of tissue destruction. The serum and urine AK activity are normally very low (as previously reported). These studies were carried out to determine the usefulness of measuring the appearance of urinary AK activity in the detection of renal transplant crisis. Serum creatinine determinations, the results of nuclear medicine studies and other clinical evidence, were compared with urinary AK activity during renal transplant crises. As a result of these studies, a model is proposed which assesses the reliability of predictors of success or failure of kidney transplants by step-wise discriminant function analysis using adenylate kinase and creatinine clearance measurements of urine to separate patients into two groups based on whether the kidney transplant was removed or retained postoperatively. The effectiveness of the predictors used in the analyses changed with respect to their importance during nonhomogeneous time intervals of analysis. PMID- 6279018 TI - Directed mutagenesis. PMID- 6279019 TI - Simian virus 40 as a eukaryotic cloning vehicle. PMID- 6279020 TI - Transposable elements in prokaryotes. PMID- 6279021 TI - [Formation of stable Hfr strains in R factor RP1 integration with the chromosome of E. coli K12 recA and their use for R-plasmid selection]. AB - Analysis of thermoindependent derivatives of E. coli K12 JC1553 recA (p VD1) carrying a replication thermostable mutant pVD1 of R factor RP1 IncP Ap Km Tc showed that formation of about 5 per cent of them was associated with stable integration of the plasmid with the bacterial chromosome. The respective bacteria had the following features: (1) preserved all the markers of plasmid pVD1, (2) according to the data of the electrophoretic analysis had no extrachromosomal DNA on prolonged cultivation under nonselective conditions, (3) were effective donors of the chromosomal genes, (4) had a low rate of the plasmid marker transfer on crossing with R- recipient. The latter feature was suggested to be used as a test for identification of stable Hfr strains. Investigation of the properties of the transconjugants obtained on crossing of stable Hfr strains with R-recipients rec+ showed that same of them had plasmid DNA with a higher molecular mass as compared to that of plasmid pVD1 DNA. The presence of this DNA was connected with formation of R' plasmid as a result of an irregular exclusion of plasmid pVD1 from the chromosome of stable Hfr bacteria. On the basis of the results a simple method was proposed for selection of R' plasmids having a number of advantages over the classical ones. The perspectives of using thermostable derivatives of RP1 for cloning the chromosome genes are discussed. PMID- 6279022 TI - Natural cell-mediated immunity to mouse mammary tumor virus and its relevance to host immune defences. PMID- 6279023 TI - Antiviral action of 5-bromo-2'-deoxyuridine and polyoma virus-specific RNA synthesis. AB - The influence of 5-bromo-2'-deoxyuridine on the synthesis of polyoma virus specific RNA in mouse embryo cells at a late phase of infection was assayed by molecular hybridization. The effect of the analogue on virus yields in terms of infectivity (p.f.u.) and haemagglutination activity (h.a.u.) was also quantitated. 12.68-126.80 micrograms/ml bromodeoxyuridine inhibited viral RNA synthesis to 15-10%, h.a.u. titers of virus yield to 50-10%, p.f.u. titers of virus yield to 0.10-0.01%, and p.f.u./h.a.u. ratios to 1.0-0.1% of the respective controls. The inhibition of viral RNA synthesis was reversed by thymidine. Since i) bromodeoxyuridine inhibited viral RNA synthesis and h.a.u. titers only partially and to a different extent, and ii) it reduced p.f.u. titers and p.f.u./h.a.u. ratios by several orders, it is likely that viral RNA synthesis represents only one of the sites of the antiviral action of the analogue. PMID- 6279024 TI - Commentary on "The current status of the hepatostatic theory of food intake control. PMID- 6279025 TI - Hepatic glucoreceptors do exist but do not control food intake. PMID- 6279026 TI - Neurophysiological evidence for hepatic glucose-sensitive afferents. Commentary on "The current status of hepatic theory of food intake control". PMID- 6279027 TI - On hepatic involvement in the short-term regulation of food ingestion. PMID- 6279028 TI - Reply to commentary on "Current status of the hepatostatic theory of food intake control". PMID- 6279029 TI - A diurnal variation in the satiating potency of cholecystokinin in the rat. PMID- 6279030 TI - Endorphinergic mechanisms in the control of food and water intake. PMID- 6279031 TI - Endogenous radiolabeling of enterotoxin from Clostridium perfringens type A on a defined medium. AB - Four enterotoxin-positive strains of Clostridium perfringens were tested for sporulation and enterotoxin production on defined media. The medium described by Sacks and Thompson (Appl. Environ. Microbiol. 35:405-409, 1978) gave the highest enterotoxin production and was selected for the production of endogenously labeled enterotoxin. The specific radioactivity of the enterotoxin was 16,000 dpm/microgram when the tritiated amino acids were added to the growth medium just before the inoculum. Addition of the radioactive amino acids during the growth period gave consistently lower specific radioactivity. When the enterotoxin was produced on the medium described by Duncan and Strong (Appl. Microbiol. 16:82-89, 1968), the highest specific radioactivity of the enterotoxin was found when the radioactive amino acids were added to the growth medium 4 h after inoculation. In this case, the specific activity of the enterotoxin was 10,000 dpm/microgram. PMID- 6279032 TI - Pathology quiz case 2: syringocystadenoma papilliferum. PMID- 6279033 TI - Ketoconazole. PMID- 6279034 TI - Serum angiotensin I-converting enzyme level in patients with cutaneous sarcoidal granulomas. AB - In a previous study we found that only half of those patients presenting with cutaneous sarcoidal granulomas have evidence of systemic involvement. The current study was designed to determine whether abnormal angiotensin-converting enzyme (ACE) levels were predictive of multisystem disease. Serum ACE levels were determined in 15 patients with active cutaneous sarcoidal granulomas. The ACE levels were elevated in ten of the patients but did clearly differentiate those with systemic involvement. Three of the six patients with disease localized to the skin had elevated ACE levels, whereas seven of the nine patients with systemic disease had elevated ACE levels. The ACE levels did not correlate with the extent of cutaneous disease, or any individual or combined system involvement. A normal ACE level cannot be used to rule out a diagnosis of sarcoidosis and, conversely, an abnormal level does not confirm multisystem involvement. PMID- 6279035 TI - Negative immunoperoxidase staining for lysozyme in nodular subepidermal fibrosis. AB - Nodular subepidermal fibrosis (NSF) is a clinical entity, the histogenetic origins of which remain unclear. More than 200 such lesions were examined with light microscopy and subdivided into four types based on their relative degree of cellularity. Five examples of each subtype were stained for lysozyme with the peroxidase-antiperoxidase technique. None of the 20 lesions contained cells with lysozyme. We conclude that the basic cell type in NSF is not lysozyme-containing macrophage; it is a different cell, perhaps one of mesenchymal origin. PMID- 6279036 TI - Cylindroma and eccrine spiradenoma coexistent in the same lesion. AB - Two patients complained of having a tender nodule. One patient had the nodule on the scalp and the other on the face. Each nodule contained a cylindroma and an adjacent eccrine spiradenoma. While eccrine spiradenoma is of eccrine derivation, the origin of cylindroma is controversial because of variable histochemical and enzyme histochemical findings. The presence of cylindroma and eccrine spiradenoma in one clinical nodule contributes circumstantial evidence for the similar derivation of both tumors. PMID- 6279037 TI - Faecal excretion of alpha-1-antitrypsin in acute diarrhoea. AB - High levels of faecal alpha-1-antitrypsin were found in children suffering from acute rotavirus diarrhoea and in children with diarrhoea in whom no microbial pathogen could be established. The results suggest that transient protein-losing enteropathy is common in acute childhood diarrhoea and may influence the outcome of the disease. PMID- 6279039 TI - Granular cell tumor of larynx and bronchus. PMID- 6279038 TI - A prospective study of hepatic imaging in the detection of metastatic disease. AB - This prospective study of 80 patients compared the results of liver scintiscan, ultrasound, and CT scan to the objective findings recorded at laparotomy. Analysis of these data revealed no significant differences in sensitivity, specificity, or accuracy among these three imaging procedures, although a trend toward increased accuracy of the CT scan was noted. In a composite analysis, the accuracy was not improved by combining two or all three of these examinations. A lesion by lesion analysis revealed the inability of any of the three currently used techniques to accurately detect lesions less than 3 cm in diameter. PMID- 6279040 TI - Delta-9-tetrahydrocannabinol: its effect on hypothalamo-pituitary system in male rats. AB - Subcutaneous administration of delta-9-tetrahydrocannabinol (delta 9-THC) in adult male rats caused a decrease in serum luteinizing hormone (LH) levels with unchanged serum prolactin, pituitary LH and pituitary prolactin content. Response of pituitary to in vitro gonadotropin releasing hormone (GnRH) remained unaltered while the response to in vivo GnRH treatment was markedly increased with the drug indicating the pituitary to be functionally normal. Differences in the in vitro and in vivo response could be due to the endogenous steroid levels. The hypothalamic LH-RH content concommitantly increased. Delta-9-tetrahydrocannabinol may inhibit the release of luteinizing hormone releasing hormone (LHRH). PMID- 6279041 TI - Freezing of retrograde ejaculate for AIH. PMID- 6279042 TI - Influence of stimulation interval on inotropism mediated by alpha-adrenoceptors in the left atria of rabbits. AB - The inotropic effects induced by alpha-adrenoceptor stimulation were investigated in the left atria of rabbits at various stimulation rates and were compared with those by beta-adrenoceptor stimulation. Phenylephrine at a concentration of 10( 5) M in the presence of 5 X 10(-5) M sotalol more increased the contractile force with decreasing the rate of stimulation from 120 to 30 beats/min, whereas the same procedure resulted in a negative inotropic effect at low stimulation rates such as 1 beat/min. Inversion from the positive inotropism to the negative one was observed to occur in a range of 3 to 12 beats/min. Rested state contractions obtained by a stimulation rate of 1 beat per 5 min, i.e. a rest period of 300 sec, were significantly decreased by phenylephrine, while increased by 5 X 10(-9) M isoproterenol. This depressive effect of phenylephrine was antagonized by 3 X 10(-6) M phentolamine. Post rest contractions (PRC) which were elicited following a rest period of 300 sec after steady stimulation of 60 beats/min, were inconsistently affected by phenylephrine in contrast with the consistent and significant increase by isoproterenol. Of 17 preparations treated with phenylephrine, 9 showed an increase and others a decrease in PRC. However, reduction of NaCl concentration by 50% resulted in a decrease in PRC in all preparations treated with phenylephrine without affecting the increase produced by isoproterenol. These results indicate that alpha-adrenoceptor-mediated positive inotropic effect inverts to a negative one when the contractions have a long interval of rest. PMID- 6279043 TI - Direct and indirect actions of 5-hydroxytryptamine in the rat anococcygeus muscle. AB - The contractile effect of 5-hydroxytryptamine on the rat isolated anococcygeus muscle was analysed. 5-Hydroxytryptamine (10(-6) g ml-1 - 1.6 X 10(-5) g ml-1) concentration-dependently contracted the anococcygeus muscle. The contractions were not competitively antagonized by methysergide (2.5 X 10(-8) g ml-1 - 10(-6) g ml-1) as the contractile response to higher concentrations of 5 hydroxytryptamine were not much reduced. The methysergide-resistant contractions to 5-hydroxytryptamine were not reduced by hyoscine (6 X 10(-7) g ml-1), tetrodotoxin (2 X 10(-7) g ml-1), but were completely abolished by phentolamine (3 X 10(-7) g ml-1). 5-Hydroxytryptamine-induced contractions were considerably reduced but not completely abolished by reserpinization. It is suggested that 5 hydroxytryptamine contracted the rat anococcygeus muscle directly (by activating tryptaminergic 'D' receptors) and indirectly by releasing noradrenaline at the nerve endings. PMID- 6279044 TI - Cardiostimulating effect of sodium linoleate and its influence on effects of cardiac glycosides. PMID- 6279045 TI - Calcium channel blockers for cardiovascular disorders. PMID- 6279046 TI - Fasting hypoglycemia in adults. PMID- 6279047 TI - How close to the theoretical diffusion limit do bacterial uptake systems function? AB - Using a 10 cm flow-through cuvette in a high precision spectrophotometer linked to a mini-computer, the growth rate dependence of Escherichia coli on glucose concentration has been studied. The specific growth rate vs bacterial mass of single cultures consuming small amounts of glucose was followed. The data were analyzed with the computer programs described previously. For neither batch nor chemostat-cultured organisms did growth follow the Monod growth law. Rather, the growth rate vs residual glucose concentration has an almost abrupt change in slope, indicative of a passive diffusion barrier prior to an uptake system possessing hyperbolic dependency. Calculations showed that the diffusion through the outer membrane via the porin channels could quantitatively account for the deviations from hyperbolic dependency. Long term chemostat culture alters the bacteria so that the maximum specific growth rate is reduced, but the initial dependence on glucose concentration is increased approaching more closely the theoretical limit. Therefore there was both a change inthe outer membrane channels and the uptake activity of the cytoplasmic membrane. PMID- 6279048 TI - Auditory brainstem evoked responses in autistic children. AB - Auditory brainstem evoked responses (ABRs) were studied in 16 autistic children. Three children had severe delays in wave I latency, indicating defective functioning of the peripheral auditory pathway. The remaining subjects also had delayed wave I latency but only for right ear stimulation at the lowest stimulus intensity. Eight autistic children (and no control subjects) had ABR transmission time values 3 SDs beyond the normal mean, suggesting auditory processing defects peripheral to or within the brainstem auditory pathway. These findings (1) may have no causal relationship to the child's autistic handicaps, (2) may represent distortions in auditory input that impair the learning of language, and (3) may reflect an earlier state in which abnormal input directly caused maldevelopment of forebrain systems necessary for language and cognitive function. PMID- 6279049 TI - Impaired presynaptic regulation of norepinephrine in schizophrenia. Effects of clonidine in schizophrenic patients and normal controls. AB - Recent studies have found elevated levels of norepinephrine (NE) in CSF and brain specimens from schizophrenic patients. Presynaptic inhibitory alpha 2-adrenergic receptors regulate NE release in the brain. The hypothesis that the functional sensitivity of this presynaptic regulation of NE is impaired in schizophrenia was tested by evaluating, in schizophrenic patients and age-matched normal controls, the ability of clonidine, an alpha 2 agonist, to lower plasma levels of the NE metabolite 3-methoxy-4-hydroxyphenylglycol (MHPG) and to lower blood pressure (BP). Clonidine produced a significant decrease in plasma MHPG levels in the normal control group, but did not lower plasma MHPG levels in the schizophrenic patients. Clonidine decreased BP equally in both groups. These results suggest that there is a functional subsensitivity of the inhibitory presynaptic alpha 2 adrenergic receptor in schizophrenia, which may relate to an impaired regulation of NE turnover. PMID- 6279050 TI - Adrenergic receptor sensitivity in depression. Effects of clonidine in depressed patients and healthy subjects. AB - Several recent investigations have raised the possibility that the sensitivity of alpha 2-adrenergic receptor may be of etiologic importance in depression. To assess whether abnormalities in presynaptic alpha 2-adrenergic receptor exist in depressed patients not taking drugs, the effects of an alpha 2 agonist, clonidine, on plasma 3-methoxy-4-hydroxyphenelethyleneglycol (MHPG) and on blood pressure (BP) were evaluated in 15 depressed patients and 12 healthy controls of similar age. The ability of clonidine to increase growth hormone (GH) secretion was also assessed. The effect of clonidine on plasma MHPG and BP was not different between the depressed patients and controls. However, the GH response to clonidine was blunted in the depressed patients. These results suggest that in depression (1) the sensitivity of the presynaptic alpha 2-adrenergic receptor is not abnormal, and (2) the sensitivity of postsynaptic adrenergic receptors may be decreased. PMID- 6279051 TI - [Viruses as co-factors in chemical carcinogenesis (author's transl)]. PMID- 6279052 TI - [Phorbol ester-induced expression of primate retroviruses (author's transl)]. AB - Addition of the tumor promoter 12-0-Tetradecanoyl-phorbol-13-acetate (TPA) to the growth medium of certain human cell cultures persistently infected with simian retroviruses of type C (BaEV, SSV) or type D (MPMV) or a human cell line-derived type D isolate (PMFV), respectively, resulted in a considerable but transient stimulation of virus production. Enhanced virus expression was paralleled by striking morphological alterations of the cells. Among four infected cell types tested so far, ony one (A 204) failed to respond to TPA with significant virus stimulation or altered morphology. PMID- 6279053 TI - Dissociation between hepatosplenic and marrow iron in liver cirrhosis. AB - Histochemical study of tissue iron in the various parts of the reticuloendothelial system in 15 subjects with cirrhosis was performed. Stainable iron in the liver and spleen sections, generally in large quantities, was found in 13 of 15 cases. Paradoxical association of hepatosplenic siderosis with depletion of marrow iron reserves was observed in seven subjects. In all seven, gross gastrointestinal blood loss had occurred during life and the source of bleeding from one or more anatomic lesions was identified at the time of autopsy. Depletion of marrow iron reserves in these seven subjects can be attributed to two factors: first, the available marrow iron stores were used for erythropoiesis; second, the ample hepatosplenic iron deposits could not be mobilized for transport to the bone marrow. Thus, hepatosplenic iron in cases of cirrhosis seems to be sequestered from its normal metabolic pathways. PMID- 6279054 TI - Uncoating-like modification of poliovirus capsid resulting from the cooperative effects of subfreezing temperature and submolar concentrations of urea. AB - Inactivation of poliovirus at subfreezing temperature in the presence of unusually low concentrations of urea (less than or equal to 0.5 M) was investigated. Whereas serotypes 1 and 2 are very sensitive, type 3 is resistant. Inactivation cannot be attributed to concentration of solutes since temperature must be reduced below -13 degrees C for loss of infectivity. Characteristics of the inactivated virion are similar to those of virions in the early stages of uncoating in HeLa cells, viz., loss of infectivity, sensitivity to proteases and detergents, change in isoelectric point, retention of intact genome, and in some instances, loss of VP4. The molecular basis for inactivation is considered to be dissociation of water bound to capsid proteins thereby causing irreversible denaturation of native tertiary structure. The results of this study are discussed in terms of their relevance to the early stages of uncoating in vivo. PMID- 6279055 TI - The genome of orf virus: restriction endonuclease analysis of viral DNA isolated from lesions of orf in sheep. AB - The purification of orf virus directly from scab material from clinical cases of orf in sheep and restriction endonuclease analysis of the viral DNA is described. Between 7 x 10(9) and 1.6 x 10(11) virus particles, and 0.7 to 22.8 micrograms of viral DNA could be recovered from 1g of scab material. Considerable heterogeneity was observed between different field isolates when restriction endonuclease digests of orf DNA were compared by gel electrophoresis. It was also shown, for two isolates, that these fragment patterns did not change after plaque purification and passage in cell culture. It is suggested that restriction endonuclease analysis of viral DNA offers a convenient method of identification of isolates of orf virus. The molecular weight of orf DNA was determined and found to be 88.8 x 10(6). PMID- 6279056 TI - Electron microscopic studies of bovine viral diarrhea virus in tissues of diseased calves and in cell cultures. AB - Pathomorphological studies by electron microscopy (EM) were carried out on the intestines and lymphoid tissues, the buffy coat cells and cultured lymphocytes from calves suffering from mucosal disease (MD). This led to the detection of particles, 45--55 nm in diameter, within characteristic vesicular structures. As these findings coincided with the isolation of bovine viral diarrhea virus (BVDV) from the same tissues and demonstration of BVDV-antigen by immunocytochemical techniques in corresponding samples, the particles were tentatively identified as the BVDV. A detailed study of in vitro infected bovine cell cultures corroborated this supposition and contributed to a conjectural evaluation of the viral morphogenesis. It revealed a difference from the morphogenesis of most other togaviruses, as the presumed virions were assembled within smooth-membraned vesicles, formed during the infection. Thus, in the material examined, a budding process was not involved in the development of BVDV. PMID- 6279057 TI - Immunogenicity of a subviral herpes simplex type 1 preparation: reduction of recurrent disease in mice. AB - Immunization of mice with a relatively small dose of subviral herpes simplex type 1 vaccine used prior to homotypic virus infection reduced the frequency of recurrent disease. However, the same vaccine dose administered after virus infection did not result in significant reduction of recurrences. PMID- 6279058 TI - The effect of herpesvirus infection on ribosomal RNA synthesis and on nucleolar size and number in HeLa cells. AB - The effects of infection by the herpesvirus pseudorabies virus on the nucleolus and on ribosomal RNA synthesis in HeLa cells were examined. The size of individual nucleoli and the total nucleolar content per cell are reduced to approximately 35 per cent of normal by 6 hours of infection. The proportion of cells containing one nucleolus rises from 10 to 50 per cent in the same time. The synthesis of ribosomal precursor RNA declines to approximately 40 per cent in this period. PMID- 6279059 TI - Herpes simplex virus infection of isolated autonomic neurons in culture: viral replication and spread in a neuronal network. AB - Cultures of isolated neurons, derived from the superior cervical ganglion (SCG) of the newborn rat and maintained in the absence of nonneuronal cells, were infected with herpes simplex virus (HSV) type 1. By phase-contrast microscopy, including time-lapse cinematography, cytopathologic changes appeared first in neuronal cell bodies and only approximately 24 hours later were axonal abnormalities detectable. Despite low yields of viral progeny, infection spread readily within the two-dimensional network of neurons and their processes. Immunoperoxidase staining for viral antigens confirmed the replication and spread of virus and revealed that antigen extended along axons during infection. Antiviral antibody added to the overlay medium slowed but did not prevent the spread of infection, indicating that virus passed from neuron to neuron over axonal pathways. Despite alteration of neuronal macromolecular synthesis early in infection, axonal transport is apparently preserved long enough to allow propagation of virus over interconnecting neural pathways. PMID- 6279060 TI - The structure of the Rous sarcoma virus glycoprotein complex: in vitro phenotypic mixing of rous sarcoma virus. AB - Gp 85 the major envelope glycoprotein of Rous sarcoma virus can be released from intact by virus treatment with 2-mercaptoethanol. Attempts were made to reassociate released gp 85 to 2-mercaptoethanol treated virus. In order to study the biological activity and to analyse the reaction product with biochemical methods gp 85 from a subgroup different from the recipient virus was used. Successful in vitro phenotypic mixing could be shown. The biochemical analysis, however, revealed that no reassociation of gp 85 and gp 35 could be obtained. The biological activity was due to a strong association of whole gp 85-gp 35 molecules derived from the gp 85 preparation to the recipient virus. These molecules were present as a minor contamination in the gp 85 preparation. PMID- 6279062 TI - A secretory product of human monocyte-derived macrophages stimulates low density lipoprotein receptor activity in arterial smooth muscle cells and skin fibroblasts. AB - The ability of macrophages to influence the metabolism of native low density lipoprotein by arterial smooth muscle cells was evaluated using cultured human monocyte-derived macrophages. Macrophage-conditioned medium stimulated the binding and degradation of low density lipoprotein by cultured arterial smooth muscle cells and skin fibroblasts. Sterol synthesis also was stimulated by macrophage-conditioned medium as was cholesterol esterification in the presence of high concentrations of low density lipoprotein. These findings suggest that macrophages secrete a factor that enhances the activity of the low density lipoprotein receptor. Low density lipoprotein degradation by arterial smooth muscle cells also was enhanced by macrophage-conditioned medium in the presence of high concentrations of low density lipoproteins in the medium. The macrophage factor that stimulates low density lipoprotein metabolism is stable to freezing, is inactivated by acid hydrolysis, tryptic digestion, and boiling, and is of large molecular weight (greater than 12,000 to 14,000 daltons). Modulation of arterial smooth muscle cell metabolism of low density lipoprotein by a macrophage secretory product may be of importance in the pathogenesis of atherosclerosis. PMID- 6279061 TI - Ocular effects of topical administration of delta 9-tetrahydrocannabinol in man. AB - delta 9-Tetrahydrocannabinol (THC) or vehicle alone was applied topically to one eye of normal paid volunteers. Ocular and systemic toxic effects and intraocular responses were measured in different series. Toxicity was limited to minor conjunctival injection that was of short (less than 60 minutes) duration and occurred with both drug and vehicle alone. Subjective responses indicated a sensation of minor burning and/or tearing. No fall in intraocular pressure was found. A small (1 mm) but significant mydriasis occurred in both the treated eye and untreated eye and was not drug related. Single-drop administration of delta 9 THC did not, therefore, cause any significant ocular irritation or reduce IOP. PMID- 6279063 TI - An outbreak of infectious bursal disease among chickens between 16 and 20 weeks old. AB - Infectious bursal disease (IBD) was diagnosed in a flock of 1,031 broilers and cockerels aged between 16 and 20 weeks. Affected birds passed whitish, watery feces. On postmortem examination, the bursa of Fabricius (bursa) was enlarged and the kidney tubules were well distended. Histopathological sections of the bursa were characterized by edema, destruction of lymphocytes, and heterophilic infiltration. Spread was rapid, and the average mortality rate was 3.5%. Bursal homogenates from dead chicks produced precipitation lines with known IBD antiserum in the agar-gel diffusion precipitation test. Sera collected from surviving chicks 10 days after the onset of the outbreak also gave precipitation lines with known IBD virus antigen. Fresh bursal homogenates from dead chicks administered intraconjunctivally to susceptible chicks exhibited typical IBD. PMID- 6279064 TI - Diagnosis of infectious bronchitis (IB) by examination of tracheal mucus for IB precipitating antigens. AB - The use of tracheal exudates for detecting precipitating infectious bronchitis (IB) antigen by the agar-gel precipitin (AGP) technique is described. Of 47 experimentally infected tracheas, 24 gave positive precipitin lines between the second and 12th day postinfection; 9 out of 18 tracheas from birds from natural outbreaks of IB were also positive. Tests can be read within 18-24 hr. For best results, several antisera or an antiserum at various concentrations should be used. PMID- 6279065 TI - Localization of viral protein in avian-encephalomyelitis-virus-infected hens. AB - Eighteen 2-year-old hens were infected orally with wild avian encephalomyelitis virus and examined using the immunofluorescent-antibody technique for 30 days. Viral antigen was detected in 10 organ tissues (liver, spleen, kidney, pancreas, and ovary) in 7 birds during the first half of the observation period and sporadically in alimentary tracts of 12 birds throughout this period, especially the lower parts of the intestines, but it was not detected in other organ tissues. Thirteen test-positive hens showed more localization of the antigen than baby chicks of other studies did. PMID- 6279067 TI - 1980 P. P. Levine Award (Dr. R. L. Witter). PMID- 6279066 TI - Comparison of precipitin antibodies and virus-neutralizing antibodies to infectious bursal disease virus. AB - Three hundred twenty-two serum samples from commercial pullets and multiplier breeders were analyzed for agar-gel precipitin (AGP) antibodies and virus neutralizing (VN) antibodies to infectious bursal disease virus. Two hundred thirty-four of these sera were AGP-positive, and 88 were AGP-negative. The geometric mean of the reciprocal of the VN titers for the AGP-positive sera was 208.7, and 232 (99.1%) had a VN titer of 1:16 or greater. In contrast, the geometric mean of the reciprocal of the VN titers for the AGP-negative sera was 6.1, but 53 (60.2%) had a VN titer ranging from 1:4 to 1:256. When the AGP test was compared with the VN test, the sensitivity and specificity, respectively, of the AGP test were 81.5% and 100%. PMID- 6279068 TI - Characterization of two strains of fowl adenovirus type 1. AB - An adenovirus isolated from the trachea of a chicken with respiratory disease was compared with an isolate obtained from the trachea of a clinically normal chicken. The isolates caused similar respiratory disease experimentally but differed in plaque morphology and fluorescent-antibody reactions in infected chick embryos. PMID- 6279069 TI - Vertical transmission of duck plague virus (DPV) by apparently healthy DPV carrier waterfowl. AB - Duck plague virus (DPV) was transmitted vertically in muscovy, pekin, and mallard ducks that were persistently infected with the LA-SD-73, MSN-WI-77, or CO-WI-73 isolates of DPV. The effects of vertical transmission on the fertility and hatchability of eggs laid by DPV carrier ducks varied with the DPV isolate and duck species. Fertility was reduced significantly only in eggs laid by MSN-WI-77 virus carrier pekin and muscovy ducks. The hatchability of eggs laid by DPV carrier mallards and muscovies was significantly reduced from that of uninfected control ducks. All ducklings tested that hatched from eggs laid by DPV carrier waterfowl shed DPV in the feces. The DPV carrier ducklings shed DPV in small amounts. Vertical transmission of DPV in domestic flocks can lower fertility and hatchability. In wild waterfowl, vertical transmission may be a means of virus perpetuation from generation to generation. PMID- 6279070 TI - Use of a constant-virus diluting-serum microneutralization technique for the detection and quantification of infectious bronchitis virus antibodies. AB - A constant-virus diluting-serum microneutralization test (CVMNT) for avian infectious bronchits virus (IBV) was evaluated for both reliability and repeatability. The virus used in the assay was a chick kidney (CK) cell-adapted strain, the Beaudette strain (IBV 42). Sera tested were from 24-week-old broiler breeder chickens that had been vaccinated 3 times from a combination vaccine of Newcastle disease virus (NDV) and IBV. Test results were not repeatable or comparable when the same sera were tested on different days, but test results were repeatable and comparable when the sera were tested on the same day. Differences in virus titer at the different times that tests were performed appeared to cause the variation in test results. A comparison was made between the CVMNT and a constant-serum diluting-virus microneutralization test (CSMNT). The CVMNT was able to detect differences in flock antibody titers that the CSMNT could not. PMID- 6279071 TI - In vitro infection of fractionated chicken lymphocytes by infectious bursal disease virus. AB - Lymphocytes from bursa of Fabricius and thymus of chickens were purified and separated into the three cell subsets--T, B, and null cells--by the techniques of nylon fiber columns and cytotoxicity tests. The in vitro susceptibility of the fractionated lymphocytes to a virulent strain of infectious bursal disease virus (IBDV) was studied by using immunofluorescence as the infection criterion. B cells were highly susceptible. By contrast, T cells and null cells were insusceptible to infection by IBDV. The relationship between the target cells for virus infection and those B cells that possessed surface immunoglobulin (SIg) was tested. B cells were further divided into SIg(M)- and SIg(G)-bearing cells by immunoadsorbent columns employing anti-immunoglobulin M(IgM) (mu-specific) or anti-IgG (gamma-specific) sera coated with Sephadex. The SIg(M)-bearing cells were highly susceptible. These results suggest strongly that SIg(M)-bearing B cells were the target cells for infection by IBDV. PMID- 6279072 TI - A simple in vitro differentiation between turkey herpesvirus and Marek's disease virus. AB - The growth and plaque formation by turkey herpesvirus (HVT) amd Marek's disease herpesvirus (MDHV) were examined in QT35 cells, a continuous fibroblast cell line derived from chemically induced tumors of Japanese quail. HVT grew and formed plaques consistently in QT35 cells when inoculated with cell-culture-propagated virus or peripheral mononuclear leukocytes (PML) from chickens that had been inoculated with HVT. Both oncogenic and nononcogenic strains of MDHV, however, failed to grow and induced neither plaques nor cytopathic effects in QT35 cells, whether inoculated with cell-culture-grown virus or heavily infected PML. When PML from chickens infected with both HVT and MDHV were assayed, only HVT plaques had developed, despite the presence in the inocula of high levels of MDHV with less HVT. The QT35 cell line provides a simple in vitro system for differentiating between HVT and MDHV and for selective isolation and identification of HVT from chickens infected with both HVT and MDHV. PMID- 6279073 TI - Histopathology of avian adenovirus group II splenomegaly of chickens. AB - Chickens were infected with an avian adenovirus group II isolate previously obtained from chickens exhibiting splenomegaly in commercial broiler flocks. The isolate was inoculated orally in 6-week-old experimental chickens, which were euthanatized and necropsied 6 days postinoculation. The primary gross lesions found were splenomegaly and splenic mottling. Numerous tissues from 12 chickens were taken for histologic evaluation. Histologic lesions included splenic reticuloendothelial cell hyperplasia with intranuclear inclusions. Lymphocytic degeneration was seen in the lungs of most chickens examined. Lung hemorrhage and edema with endothelial disruption and congestion of pulmonary arterioles were found less frequently. The splenic lesions in the chickens were similar to those seen in turkeys naturally infected with hemorrhagic enteritis, and the lung lesions resembled those seen in pheasants naturally infected with marble spleen disease. PMID- 6279074 TI - Canary pox vaccination with live embryo-attenuated virus. AB - A live-virus embryo-propagated canary pox vaccine was developed that can be safely and effectively applied by a single needle wing-web stab. Reactions to vaccination with passages lower than the 72nd passage were sometimes excessive in proliferative response. The 72nd and 100th embryo passages produced a more restricted lesion at the site of inoculation. The vaccine does not cause a systemic reaction or metastasis from the inoculation site. PMID- 6279075 TI - Natural killer cell activity in chickens exposed to Marek's disease virus: inhibition of activity in susceptible chickens and enhancement of activity in resistant and vaccinated chickens. AB - Chickens of 2 genetic lines (lines P and N) were inoculated with a pathogenic strain of Marek's disease (MD) virus (MDV) and chronologically examined for disease response and natural killer (NK) cell expression. The NK cell reactivity was assayed in an in vitro cytotoxicity assay in which effector cells from the spleen of test chickens were reacted with 51Cr-labeled LSCC-RP9 target cells. Chickens of line P developed progressive debilitating disease and a high incidence of gross tumors and death. The NK cell reactivity of line-P chickens infected with MDV was significantly lower than that of uninfected control hatchmates. In contrast, NK cell levels were significantly elevated in MDV inoculated line-N chickens that were resistant to MD and in chickens of lines P or N that had been inoculated with herpesvirus of turkeys (HVT). NK cell levels were also elevated in line P if chickens were vaccinated with HVT before infection with MDV. Inhibition of NK reactivity in susceptible chickens and elevation of reactivity in naturally resistant or vaccinated chickens may indicate a role for the NK cell system in regulating resistance to MD. PMID- 6279076 TI - Characteristics of apparent derivatives of the 2512 strain of infectious bursal disease virus when used as vaccines. AB - The 2512 infectious bursal disease virus (IBDV) strain maintained in the authors' laboratory was compared with apparent 2512 IBDV isolates designated I-2512, P 2512, and H-2512. The latter three viruses were obtained from different sources and, ostensibly, had their origin in the Purdue laboratory. Their effects on immunogenicity, transmissibility, pathogenicity, and cell cultures varied. One of these isolates was said to be only 2 embryo passages higher than the original seed virus in our laboratory. Included in the study, also for comparison, was a cell-cultured-adapted 2512-cloned attenuated virus. The findings emphasize changes that may occur in the identity of a virus from manipulation, mutation, storage, errors in labeling, or other factors. These characteristics should be identified if the virus is to be used as a vaccine, in research, or for other purposes. The need for well-characterized reference strains in repositories is discussed relative to their importance in potential vaccine research and development. PMID- 6279077 TI - Induction of lymphomas and tumor antigen by Marek's disease virus in turkeys. AB - Pathogenic Marek's disease (MD) virus (MDV) induced a high incidence of mortality and gross and microscopic lesions in turkey poults. Inoculated turkey poults became persistently viremic with MDV, although the levels of detectable circulating MDV were generally lower in turkeys than in similarly inoculated chickens. The early lytic phase of MD characterized by lymphoid cell destruction and the appearance of viral antigen in lymphoid organ was not as prominent in turkeys as in chickens. Gross MD lesions in turkeys were most prevalent in liver and spleen; peripheral nerves were involved infrequently. MD tumors in turkeys contained cells that reacted with antiserum prepared against chicken MD-tumor associated surface antigen (MATSA); this result indicated that MDV induced cellular transformation in turkeys and the tumor-associated antigen in chicken and turkeys cross-reacted. Several in vitro-propagating B-cell lines were developed from the turkey lymphomas. As in chickens, MD in turkeys also resulted in immunodepression. Circulating lymphocytes from turkeys that eventually died of MD or had gross tumors at the end of the experiment were deficient in mitogenic response to Concanavalin A. PMID- 6279078 TI - Growth and infectivity titration of virulent infectious bursal disease virus in established cell lines from lymphoid leukosis tumors. AB - Lymphoblastoid cell lines were investigated for susceptibility to virulent (wild) infectious bursal disease virus (IBDV) and attenuated IBDV by detection of fluorescent antigen in inoculated cells. LSCC-BK3 and LSCC-CU10 cells were susceptible to both types of IBDV; LSCC-1104-B-1 line was susceptible to only the attenuated IBDV. However, 2 lymphoid leukosis and 2 Marek's disease cell lines were refractory to growth of any type of IBDV. When the J1 strain of virulent IBDV was inoculated into LSCC-BK3 cells, intracellular and extracellular viruses began to increase logarithmically between 4 and 8 hr postinoculation and reached 10(6.3) and 10(7.3) TCID50/0.1 ml, respectively, at 72 hr. Infectivity titration and serum-neutralization tests of virulent IBDV were possible with LSCC-BK3 cells. PMID- 6279079 TI - Pathogenesis of infectious bursal disease in embryonally bursectomized chickens. AB - The pathogenesis of infectious bursal disease (IBD) in intact chickens was compared with pathogenesis in chickens that had undergone embryonal bursectomy (EBX chickens), which were challenged at either 2 or 6 weeks of age. All EBX chickens were free of bursa remnants, and those challenged at 6 weeks of age failed to develop primary and secondary antibody responses to sheep red blood cells and bovine serum albumin. A direct fluorescent-antibody technique was used to study the course of infection in the bursa of Fabricius, spleen, thymus, cecal tonsils, and kidneys. EBX delayed but did not prevent the appearance of IBD infected cells. Virus-positive cells were detected in the thymus of EBX chickens up to 18 days postinfection. Significantly more EBX chickens challenged at 6 weeks of age had hemorrhagic lesion in muscles and intestinal tract than comparable intact chickens had. PMID- 6279080 TI - Perspectives for the genetic engineering of hydrocarbon oxidizing bacteria. PMID- 6279081 TI - Models for genetic manipulation of Actinomycetes. PMID- 6279082 TI - Strategies for cloning in Bacillus subtilis. PMID- 6279083 TI - Molecular cloning in the Streptococci. PMID- 6279084 TI - Genetic engineering on microorganisms for chemicals: diversity of genetic and biochemical traits of pseudomonads. PMID- 6279085 TI - Cloning vectors derived from bacterial plasmids. AB - A wide variety of plasmid cloning vectors, most of which utilize the basic Co1E1I replicon have been constructed. Utilizing these vectors, in conjunction with the newly developed techniques of gene isolation and oligonucleotide synthesis, essentially any gene which can be identified can be cloned. We anticipate that future work in this area will be directed at improving techniques for the regulated expression of cloned genes and the further development of plasmid replicons in which the copy number can be readily controlled. PMID- 6279086 TI - The alcohol dehydrogenase genes of the yeast, Saccharomyces cerevisiae: isolation, structure, and regulation. PMID- 6279087 TI - Plasmids as vectors for gene cloning: past, present, and future use. PMID- 6279088 TI - Baker's yeast: a successful industrial microorganism is now a favorable host for molecular cloning. PMID- 6279090 TI - [Chorioiditis, chorioretinitis]. PMID- 6279089 TI - Transformation of Neurospora crassa utilizing recombinant plasmid DNA. PMID- 6279091 TI - [Panuveitis]. PMID- 6279092 TI - A single serum dilution method for the quantitation of neutralizing antibodies to varicella-zoster virus. AB - A one-point serum dilution method for determination of neutralizing antibody in human sera to varicella-zoster (V-Z) virus instead of the serial serum dilution method was investigated. Focus counting was performed under a microscope on day 5 to 6 after inoculation of V-Z virus into 6-well plastic trays in which human embryonic lung cells were grown. A table was constructed to estimate the ND50 titers by the per cent reduction of the focus count from the control at only one dilution of test sera. The estimated ND50 values agreed well with those determined by the serial serum dilution method. Test sera showed a slight nonspecific reactivity at low serum dilutions, but reliable results could usually be obtained at a serum dilution of 1:8 or more. This method, which saves materials and labor, was applied to the quantification of neutralizing antibody against V-Z virus in human sera with satisfactory accuracy and reproducibility. PMID- 6279093 TI - Isoelectric-focusing patterns of cyclic nucleotide phosphodiesterase from rat heart. AB - 1. Isoelectric focusing on a flat gel bed of the rat heart cytosolic fraction resolved cyclic nucleotide phosphodiesterase activity into several forms, characterized by their substrate specificity, kinetic constants and dependence towards Ca2+ and calmodulin. A peak of pI 4.9 displayed 20 times more affinity for cyclic GMP than for cyclic AMP and was markedly inhibited by EGTA. A less substrate-specific form, only slightly sensitive to EGTA inhibition, focused at pH 5.45. Several overlapping peaks detected between pH 5.55 and pH6 specifically hydrolysed cyclic AMP, with non-Michaelian kinetics; these peaks were insensitive to Ca2+ chelation. 2. Isoelectric focusing did not dissociate enzyme-calmodulin complexes, as none of the resulting peaks was activatable by calmodulin plus Ca2+. 3. Some new information on rat cardiac phosphodiesterase is obtained with this technique, which is convenient for routine analytical studies of phosphodiesterase, as well as for preparative purposes. PMID- 6279094 TI - Interaction between non-classical beta-lactam compounds and the Zn2+-containing G and serine R61 and R39 D-alanyl-D-alanine peptidases. AB - Streptomyces albus G secretes a Zn2+-containing D-alanyl-D-alanine peptidase. Streptomyces R61 and Actinomadura R39 secrete D-alanyl-D-alanine-cleaving serine peptidases. The effect of non-classical beta-lactam antibiotics on these three model enzymes has been studied. Mecillinam, cefoxitin, quinacillin, quinacillin sulphone, clavulanate and N-formimidoylthienamycin have no effect on the Zn2+ containing enzyme. 6-Amino-penicillanic acid slowly inactivates this enzyme and 7 aminocephalosporanic acid behaves as a reversible inhibitor. Cefoxitin and N formimidoylthienamycin are potent anti-bacterial agents; they effectively inactivate the serine R39 enzyme and, to a lesser extent, the serine R61 enzyme. All the other beta-lactam compounds tested, including mecillinam, are slow inactivators of these serine enzymes. The intermediates formed between 6 aminopenicillanic acid and the R61 and R39 enzymes are long- and short-lived respectively, whereas those formed between 7-aminocephalosporanic acid and the same R61 and R39 enzymes are short- and long-lived respectively. Breakdown of the short-lived intermediates thus obtained gives rise to several ninhydrin-positive degradation products. The intermediates formed between clavulanate and the serine enzymes are long-lived. With the R39 enzyme, the inactivated complex formed in a first step undergoes subsequent monomolecular rearrangement to give rise to a second species exhibiting a high absorbance at 273 nm. PMID- 6279095 TI - The steady-state kinetics of the NADH-dependent nitrite reductase from Escherichia coli K 12. Nitrite and hydroxylamine reduction. AB - The reduction of both NO2- and hydroxylamine by the NADH-dependent nitrite reductase of Escherichia coli K 12 (EC 1.6.6.4) appears to follow Michaelis Menten kinetics over a wide range of NADH concentrations. Substrate inhibition can, however, be detected at low concentrations of the product NAD+. In addition, NAD+ displays mixed product inhibition with respect to NADH and mixed or uncompetitive inhibition with respect to hydroxylamine. These inhibition characteristics are consistent with a mechanism in which hydroxylamine binds during catalysis to a different enzyme form from that generated when NAD+ is released. The apparent maximum velocity with NADH as varied substrate increases as the NAD+ concentration increases from 0.05 to 0.7 mM with 1 mM-NO2- or 100 mM hydroxylamine as oxidized substrate. This increase is more marked for hydroxylamine reduction than for NO2- reduction. Models incorporating only one binding site for NAD can account for the variation in the Michaelis-Menten parameters for both NADH and hydroxylamine with [NAD+] for hydroxylamine reduction. According to these models, activation of the reaction occurs by reversal of an over-reduction of the enzyme by NADH. If the observed activation of the enzyme by NAD+ derives both from activation of the generation of the enzyme-hydroxylamine complex from the enzyme-NO2- complex during NO2- reduction and from activation of the reduction of the enzyme-hydroxylamine complex to form NH4+, then the variation of Vapp. for NO2- or hydroxylamine with [NAD+] is consistent with the occurrence of the same enzyme-hydroxylamine complex as an intermediate in both reactions. PMID- 6279096 TI - Pre-incubation of subcellular fractions from rat cerebral cortex inactivates protein phosphorylation. AB - Pre-incubation of various subcellular fractions from rat brain caused a significant decrease in the phosphorylation of individual polypeptides. The rate and extent of this loss of labelling was not uniform and polypeptides whose phosphorylation was independent of cyclic AMP were primarily affected, whereas substrate availability remained unaltered. It is recommended that pre-incubation effects must be carefully monitored if valid conclusions are to be made about the physiological relevance of changes in protein phosphorylation observed in vitro. PMID- 6279097 TI - The stereochemical course of phosphoryl transfer catalysed by polynucleotide kinase (bacteriophage-T4-infected Escherichia coli B). AB - Polynucleotide kinase (bacteriophage-T4-infected Escherichia coli B) catalyses the transfer of the [gamma-16O,17O,18O]phosphoryl group from 5'[gamma(S) 16O,17O,18O]ATP to 3'-AMP with inversion of configuration at the phosphorus atom. The simplest interpretation of this observation is that the [gamma 16O,17O,18O]phosphoryl group is transferred directly from ATP to the co-substrate by an 'in-line' mechanism. PMID- 6279098 TI - Protein crystals, membrane proteins and membrane lipids. Recent advances in the study of their static and dynamic structures using nuclear magnetic resonance spectroscopic techniques. PMID- 6279099 TI - Electron spin resonance studies of lipid-protein interactions in membranes. PMID- 6279100 TI - Studies of larger motions within proteins by nuclear magnetic resonance spectroscopy. PMID- 6279101 TI - Aqueous ion channels. AB - The purpose of this paper is to describe models of ion channels in membranes which are built around a central core of ordered water. The calculated characteristics of such channels will be described and compared when possible with the measured properties of real ion channels. The approach is descriptive with reference when necessary to more detailed calculations contained in two previous papers (Edmonds, 1979, 1980). To place in perspective the problems of designing realistic model mechanisms for the translocation of ions across lipid membranes, the electrostatic self energy of a charged particle is introduced at the start. The severe limitations this energy places upon the environment that an ion must experience in traversing the lipid bilayer are briefly discussed. PMID- 6279102 TI - Enzymatic phosphorylation of acyclic nucleoside analogs and correlations with antiherpetic activities. PMID- 6279103 TI - Differences in the inhibitory effect of Cd2+, Mn2+ and Al3+ on the uptake of dopamine by synaptosomes from forebrain and from striatum of the rat. PMID- 6279104 TI - Cyclic AMP response to norepinephrine in the limbic forebrain of male and female rats: effect of desipramine. PMID- 6279105 TI - Evidence for more than one type of post-junctional alpha-adrenoceptor. AB - The concept of two types of alpha-adrenoceptor, alpha 1 located on smooth muscle and mediating contraction and alpha 2 located on nerve terminals and mediating inhibition of transmitter release, has broken down. In vivo it has been shown that post-junctional receptors, with characteristics closely related to those of the alpha 2-adrenoceptors at nerve terminals, can mediate pressor responses and are, "post-junctional alpha 2-adrenoceptors". Several differences among agonists in vitro have superficial similarities to the in vivo alpha 1/alpha 2 system but do not correspond precisely and seem to point to a subdivision of post-junctional alpha 1-adrenoceptors. A preliminary hypothesis is: in vivo alpha 1 is rapid in onset, short-lived, utilises internal Ca2+, prefers alkalosis and responds to short-term stimuli such as short bursts of nerve impulses or bolus injections of catecholamines; alpha 2 is slower in onset, longer-lived, utilises external Ca2+, prefers acidosis and responds to more prolonged stimuli such as circulating catecholamines; in vitro these categories of response occur but antagonists fail to define an alpha 1/alpha 2 split, suggesting that some critical factor is missing in vitro. The implications of these trends in alpha-adrenoceptor classification are discussed in relation to current pharmacological and biochemical methods for receptor typing, to the possible physiological actions and roles of such receptors and to structure/activity relationships among agonists and antagonists. PMID- 6279106 TI - H2O2 production, modification of the glutathione status and methemoglobin formation in red blood cells exposed to diethyldithiocarbamate in vitro. AB - Human red blood cells treated with the CuZn superoxide dismutase inhibitor diethyldithiocarbamate (DDC) undergo metabolic modifications in addition to the superoxide dismutase inhibition: oxidation of the reduced glutathione (GSH) to oxidized glutathione (GSSG), methemoglobin formation, and increased hexose monophosphate shunt activity were observed. The magnitudes of these changes are dependent on the DDC concentration. Under nitrogen, only superoxide dismutase inhibition occurs. After removal of the GSH with N-ethylmaleimide, production of H2O2 can be detected by measuring the red cell catalase inhibition in the presence of 3-amino-1,2,4-triazole. H2O2 production is not altered by conversion of oxyhemoglobin to methemoglobin by sodium nitrite prior to incubation. GSH oxidation and methemoglobin formation are stopped when DDC is eliminated from the incubation medium after completion of the superoxide dismutase inhibition. These data indicate that methemoglobin formation and modification of the GSH status in red cells treated by DDC are not a direct consequence of the CuZn superoxide dismutase inhibition but are due rather to a DDC-dependent production of H2O2. PMID- 6279107 TI - Interaction of drugs with a model membrane protein. Effect of dibucaine on cytochrome oxidase proteoliposomes. AB - Cytochrome oxidase is a mitochondrial trans-membrane protein which catalyzes the vectorial transfer of electrons from cytochrome c to molecular oxygen. When the oxidase was incorporated into liposomes composed of saturated phospholipids, enzymatic activity was reduced as compared to the activity of either the isolated enzyme or the enzyme incorporated into soy bean phospholipid (asolectin) liposomes. This reduced activity probably resulted from partial replacement of retained oxidase boundary lipid with exogenously added lipid and an unfavourable orientation of a portion of the oxidase molecules for reaction with externally added substrate. On the other hand, substrate binding at the low affinity site was enhanced by incorporation of the oxidase into vesicles composed of either saturated phospholipids or asolectin. At pH 7.4 the local anesthetic dibucaine behaved as an uncompetitive inhibitor of the enzyme, while at pH 6.0 the inhibition pattern became mixed in type. Dibucaine had similar effects on both the isolated and incorporated enzyme except that, in general, the anesthetic caused less inhibition of the incorporated oxidase. It is postulated that positively charged anesthetic molecules act predominantly by competing with substrate for binding while non-charged anesthetic molecules interact with the oxidase boundary lipid to form non-productive complexes. PMID- 6279108 TI - Effects of polychlorinated biphenyls and environmental temperature on in vitro formation of benzo[a]pyrene metabolites by liver of trout (Salmo gairdneri). AB - Rainbow trout (Salmo gairdneri) held at 7 degrees and 16 degrees were given Aroclor 1254 (PCB) (10 mg/kg body wt) via intraperitoneal injections. The binding of [3H]benzo[a]pyrene (BaP) to deproteinized salmon sperm DNA was assayed (pmoles BaP equivalents per mg DNA per mg protein) using the post-mitochondrial supernatant (S 10) fractions from livers of fish at 24-168 hr after the PCB exposure. Liver enzymes from the untreated fish acclimated at 7 degrees yielded an average binding value (0.37 +/- 0.17) which was significantly greater (P less than 0.05) than the value (0.07 +/- 0.03) for untreated fish at 16 degrees. Liver supernatant fractions from PCB-induced fish acclimated at 16 degrees and sampled at 24-120 hr showed a substantial increase (P less than 0.05) in the binding (average value 2.4 +/- 1.8) compared to the value obtained with untreated fish at 16 degrees. At 24, 48 and 120 hr after the PCB treatment of fish held at 7 degrees, there was no significant increase in the binding value or extent of metabolism of BaP compared to that obtained with the untreated fish at 7 degrees. However, at 168 hr, three of four fish at 7 degrees responded to the PCB treatment with significantly (P less than 0.05) increased binding values (3.3 +/- 1.6). Chromatographic analyses of the ethyl acetate-soluble metabolites revealed that 3-hydroxy BaP and 7,8- and 9,10-dihydrodiols were the major metabolites; K region metabolites were formed in trace amounts in untreated and PCB-treated fish at both temperatures. No marked qualitative differences were observed in metabolite profiles after the PCB treatment; however, overall metabolism of BaP and production of reactive metabolites by liver enzymes were considerably (P less than 0.05) enhanced in the PCB-induced fish at both 7 degrees and 16 degrees. PMID- 6279109 TI - K+ release from rat parotid cells: an alpha 1-adrenergic mediated event. AB - To characterize alpha-adrenergic receptors in rat parotid gland tissue, 9,10 [9,10-3H(N)]-dihydro-alpha-ergocryptine ([3H]DHE) and [3H]prazosin binding to membranes and stimulated K+ release from parotid cell aggregates were examined. Prazosin (selective alpha 1-adrenergic antagonist), displacement of [3H]DHE binding from parotid membranes was biphasic, indicating the presence of both alpha 1- and alpha 2-adrenergic receptors. The numbers of alpha 1- and alpha 2 receptors were about equal. alpha 1-Adrenergic receptors were further studied by [3H]prazosin binding. [3H]Prazosin binding was a rapid, reversible, saturable and stereospecific process, with high affinity (KD = 0.38 nM) and low capacity (Bmax = 380 fmoles ligand bound/g tissue, 10.1 fmoles/mg protein) as determined by Scatchard analysis. The characteristics of [3H]prazosin binding were in good agreement with those of catecholamine-stimulated K+ release, suggesting that K+ release from rat parotid gland cells is an alpha 1-adrenergic mediated effect. PMID- 6279110 TI - Further studies on the generation of reactive oxygen species from activated anthracyclines and the relationship to cytotoxic action and cardiotoxic effects. AB - The relative ease of generation of reactive oxygen species from a series of reductively activated aglycone and sugar modified anthracyclines was compared by the extents of single strand scission in supercoiled PM2-covalently closed circular (CCC)-DNA. The electrochemical properties of these agents were used as a quantitative measure of the ease of reduction and subsequent reoxidation of the reduced species. The relationship of these processes to various biological properties of the anthracyclines, in particular to the measured cardiotoxicity of the drugs, was examined. An attempt was made to identify those structural changes which ameliorate the cardiotoxic effects measured in other test systems while permitting the expression of antitumor properties. PMID- 6279111 TI - Localisation of [3H]clonidine binding to membranes from guinea pig renal tubules. AB - The selective radioligand [3H]clonidine has been used to localise alhpa 2 adrenoceptors in guinea pig kidney. Chemical sympathectomy with 6-hydroxydopamine produced no significant change in the number of sites labeled by [3H]clonidine indicating that the majority of binding sites were not located on sympathetic nerve terminals. Binding was enhanced in membranes prepared from renal tubules and considerably reduced in preparations from glomeruli. Subcellular fractions of renal cortex revealed that binding was to plasma membranes and that the greatest binding capacity was present in the fraction rich in basal lateral membranes. It is concluded that the major concentration of renal alpha 2 adrenoceptors are present on renal tubules and that they may be localised to a particular pole of the renal tubule cell. PMID- 6279112 TI - [3H]Diazepam binding sites on rat heart and kidney. PMID- 6279113 TI - Prostaglandins and cannabis--XI. Inhibition of delta 1-tetrahydrocannabinol induced hypotension by aspirin. PMID- 6279114 TI - Pyrophosphate arthropathy in two Swedish families. AB - A clinical and radiographic survey of 110 members of 2 families with hereditary pyrophosphate arthropathy was performed. The mode of inheritance was autosomal, dominant with a variable penetrance. Twenty-two percent of the family members had joint involvement related to pyrophosphate arthropathy, 47% of those over 50 years of age had experienced acute attacks of arthritis and/or had joint calcifications. The majority of individuals with both arthritis and joint calcifications suffered from chronic pain that resulted in early retirement. A high frequency of back pain was observed, but no ankylosis or deformity. Surgery was performed for parathyroid hyperplasia on the propositus in 1 family, and several members of her family suffered from symptoms that suggested a disturbance of calcium phosphate metabolism. There were several differences between our patients and 50 cases of sporadic pyrophosphate arthropathy from the same area of Sweden. Familial cases had an earlier onset, a greater number of involved joints, and peripheral joint involvement more often. Back pain was more frequent, and calcifications of intervertebral discs were found only in the hereditary cases. PMID- 6279115 TI - Superoxide anion generation by human neutrophils exposed to monosodium urate. AB - We studied the capacities of naked and protein-coated monosodium urate (MSU) crystals to stimulate superoxide anion (O(2)) release by human polymorphonuclear leukocytes (PMN). Uncoated MSU estimated significant O(2) production by cytochalasin B-treated PMN. Precoating MSU with IgG caused an increase in mean O(2) production, whereas precoating heated MSU with serum or plasma inhibited O(2) release. Unheated MSU crystals, which activate complement to a greater extent than heated crystals, also provoked O(2) generation, an effect again abrogated by precoating with serum but not with plasma. Coincubation of unheated MSU and plasma resulted in an enhancement of O(2) generation. The results of these experiments support the hypothesis that adsorbed proteins modulate the phlogistic potential of MSU and that the surface activation of humoral mediators contributes to the local inflammatory response. PMID- 6279116 TI - Stimulation of the respiratory burst in human neutrophils by crystal phagocytosis. AB - Exposure to monosodium urate crystals (MSU) stimulated the respiratory burst of human neutrophils as measured by increased O2 consumption and the generation of superoxide radicals (O(2)). From the comparison of data derived from nitroblue tetrazolium and cytochrome C reduction (two methods of detecting O(2) release), it appears tht O(2) production in response to MSU may be compartmentalized, i.e., occur predominantly in the intracellular space. After exposure to MSU, neutrophils from patients with chronic granulomatous disease lost viability at the normal rate; thus products of the respiratory burst are not likely to be responsible for cell death. PMID- 6279118 TI - Antibody to cytomegalovirus in patients with Sjogren's syndrome. As determined by an enzyme-linked immunosorbent assay. AB - By use of enzyme-lined immunosorbent assay (ELISA), patients with Sjogren's syndrome were found to have levels of serum IgG antibody to cytomegalovirus twice those of matched control subjects, and IgM antibody levels which were three times as high. It may be relevant that features of cytomegalovirus infection are similar to those of Sjogren's syndrome. PMID- 6279117 TI - Variable response to oral angiotensin-converting-enzyme blockade in hypertensive scleroderma patients. AB - Twelve scleroderma patients with hypertension, seven of whom had malignant hypertension and renal failure of scleroderma renal crisis, were treated with captopril. The first dose lowered mean pressure in all patients by 21.3 mmHg; in 6 patients it relieved encephalopathy. Blood pressure was controlled in all patients. Two of 7 patients with scleroderma renal crisis had improvement in renal function; the 5 patients who did not have malignant hypertension improved or stabilized. Despite good pressure control, however, renal failure developed in 5 patients with scleroderma renal crisis. The data indicated that captopril is effective antihypertensive therapy in scleroderma and, when given early, may prevent renal failure and death. PMID- 6279119 TI - Cauda equina syndrome in a patient with high titer anti-RNP antibodies. PMID- 6279120 TI - Acute temporomandibular arthritis in a patient with bruxism and calcium pyrophosphate deposition disease. PMID- 6279121 TI - Electron spin resonance studies on the influence of carbocromen on mitochondria and oligomycin-sensitive mitochondrial ATPase. AB - The action of ethyl 3-(2-diethylaminoethyl)-4-methyl-2-oxo(2H)-1-benzopyran-7 yloxyacetate (carbocromen, Intensain) on the structure of mitochondria from rat liver and bovine heart and on oligomycin-sensitive (OS)-ATPase was studied using the spin labelling method (ESR). Most prominent results were found with the spin label 4-maleimido-2,2,6,6-tetramethylpiperidinooxyl. In bovine heart mitochondria ho/h-1 ratios in repeated scans increased in presence of carbocromen (20--40 mumol/l), indicating a decrease in mobility of the spin label 100. Reduction rates of spin label were comparable in both controls and carbocromen treated samples. Carbocromen under similar conditions had no effect on rat liver mitochondria. In bovine heart mitochondrial OS-ATPase a concentration-dependent increase of mobile parts of the spectra was found. The ATPase activities indicated a substrate inhibition from concentration of about 2.85 mmol ATP onward. As a working hypothesis it is proposed that in the presence of high substrate concentrations, the drug may decrease ATPase activity, an elevation of which may have fatal consequences in myocardial ischemic disease. PMID- 6279122 TI - Most biological effects of zinc deficiency corrected by gamma-linolenic acid (18: 3 omega 6) but not by linoleic acid (18: 2 omega 6). PMID- 6279123 TI - Effect of blackgram fibre on ethanol-induced hyperlipidemia in rats. AB - Rats fed a diet low in fibre and provided with ethanol for 4 weeks showed a higher concentration of cholesterol in the serum, liver and heart, but not in the aorta when compared with control rats not provided with ethanol. Animals maintained on a diet of blackgram fibre (30%) and provided with ethanol had significantly lower concentration of cholesterol in these tissues and in the aorta. The concentration of triglyceride was also raised in the serum, liver and heart in rats fed a diet low in fibre + ethanol. A diet of blackgram fibre caused a significant decrease in serum and liver triglyceride. Fecal excretion of neutral sterols and bile acids decreased in rats fed a diet low in fibre + ethanol, whereas blackgram fibre caused an increase in such fecal excretion. PMID- 6279124 TI - The fatty acids of erythrocytes of myocardial infarction patients. AB - Arachidonic acid (C20 : 4 omega 6) and eicosapentaenoic acid (C20 : 5 omega 3) are precursors of two different series of prostaglandins important in homeostasis of the cardiovascular system. The levels of these and other fatty acids have been measured in a group of 20 patients who had suffered myocardial infarction (samples taken within 12 h of infarction) and a group of 17 healthy age-matched controls using the erythrocyte as a lipid probe. There was no significant difference between the level of C20 : 4 omega 6 in patients and controls. There was however a highly significant difference in the level of the peak containing C20 : 5 omega 3 i.e. 6.60% +/- 0.29 (mean and SE of mean) for controls; 3.91 +/- 0.36 (mean and SE of mean) for patients with myocardial infarction. In each of the two groups the relationship between the levels of the two fatty acids arachidonic acid, eicosapentaenoic acid and their essential fatty acid precursors has been investigated. No significant functional relationship was found except between C18 : 3 omega 3 and C20 : 5 omega 3 in the control group. These results are discussed in relation to homeostasis and recent evidence that levels of these fatty acids can be altered by dietary manipulation. PMID- 6279126 TI - [IgM-specific neutralizing antibodies against poliovirus in cases of poliomyelitis]. PMID- 6279125 TI - The number of LDL receptors and age at myocardial infarction. PMID- 6279127 TI - [Molecular organization of erythrocyte membranes in Duchenne muscular dystrophy]. AB - Lipid-soluble spin labels were used to study lipid protein interactions in erythrocyte membranes obtained from patients with Duchenne muscular dystrophy. The temperature dependence of order parameter (Sn) and correlation time (tau c) have demonstrated alterations in the surface and in the hydrophobic core of membrane; the differences might be correlated with a depletion of extrinsic protein and of intramembrane particles. Our studies suggest a great abnormality of membrane molecular organization in Duchenne muscular dystrophy. PMID- 6279128 TI - [GH after baclofen and diazepam]. AB - It has been proposed that GABAergic receptors possess two functionally distinct, but strictly co-operating binding sites, one for GABA itself, another for benzodiazepines. The activation of GABA receptor is followed by opening of the Cl channels. This action is strengthened if both binding sites of GABA receptor are contemporaneously engaged by their respective agonists. Baclofen, a GABAergic drug, is able, in acute administration, to determine an increase of serum GH in man. The aim of this work was to control if benzodiazepines can strengthen this metabolic effect of Baclofen in vivo. Five healthy voluntary women were studied. Blood samples for serum GH evaluation were collected basally and 30, 60, 90, 120, 150, 180, 210 minutes after oral administration of 20 mg. of Baclofen. This test was repeated one week after oral administration of 12 mg. die of Diazepam. The results were analyzed for paired data. The finding show that the metabolic effect of Baclofen on GH secretion are prolonged in time after Diazepam premedication and suggest that GABA-benzodiazepines interaction is effective at hypothalamo hypophyseal level in man. PMID- 6279129 TI - [Influence, in normal subjects, of an isocaloric hyperprotein diet on cortisol, ACTH, GH and PRL response to lysine-8-vasopressin]. AB - The Lysin-8-Vasopressin test has been experimented in ten healthy subjects during normocaloric balanced diet and after hyperproteic-normocaloric diet. The levels of ACTH, Cortisol and GH are significantly more elevated after hyperproteic normocaloric diet than in basal conditions. The levels of Prolactin do not show any remarkable change. These results can indicate the increased reactivity of the diencephalon-hypophysis-adrenal axis and of the hormones connected with the mechanisms of homeostasis and stress, probably correlated to more disposable proteic material and to the metabolic effects which follow. PMID- 6279130 TI - [Adsorption of proteins by artificial mineral fibers: in vitro experiments]. AB - Since previous studies reported that in vitro some proteins and phospholipids were absorbed by asbestos fibres, namely chrysotile, in this study, man made filamentous glass fibers are been tested in vitro at the presence of proteins. The objective was to obtain evidence to evaluate whether continuous glass fibers have a behaviour similar to asbestos fibres. A sample of chrysotile fibres was used as control. Uptake of bovine serum albumin and horse spleen ferritin on these continuous glass fibres has been observed. However on glass fibres adsorbed less proteins per weight unit (22 mg/g and 12 mg/g respectively for albumin and ferritin) than asbestos chrysotile fibres (42 mg/g and 49 mg/g respectively for albumin and ferritin). PMID- 6279131 TI - [The influence, in normal subjects, of a high-protein normocaloric diet on the response of cortisol, ACTH, GH, and PRL to insulin hypoglycemia]. AB - A protein rich diet causes a remarkable increment of plasma cortisol, corticotropin and somatotropin concentration, but does not modify the plasma prolactin level; this diet, moreover, is followed by a more vivacious response to the Lysin-8-Vasopressin test. In 10 healthy voluntary subjects we have studied the hormonal behaviour during the insulin-induced hypoglycemia test in course of equilibrated diet and after 15 days of protein-rich diet. In these two experimental conditions the insulin-induced hypoglycemia test has promoted a similar increment of the four hormones. The different behaviour between the two tests -Lysin-8-Vasopressin and insulin-induced hypoglycemia- indicates that the increased hormonal levels which follow a protein-rich diet are not provoked by a generic stress effect, but by a direct stimulation of the hypothalamo-hypophyseal structures. PMID- 6279132 TI - [Cardiac metastases and neoplastic pericarditis: clinical cases]. PMID- 6279133 TI - [Effect of the cold pressor test on erythrocyte beta-adrenergic receptors in essential hypertension and healthy subjects]. PMID- 6279134 TI - Influence of medication, pain and progress in labour on plasma beta-endorphin like immunoreactivity. AB - Plasma beta-endorphin-like immunoreactivity (beta-ELI) was measured at intervals during labour in 42 patients. An increase in beta-ELI occurred during labour and increased to a maximum during the second stage. The increase in beta-ELI paralleled dilatation of the cervix. A correlation existed between the duration of labour and second stage beta-ELI in patients with labours of more than 4 h duration. Analgesic drugs were associated with changes in beta-ELI. The patients who received Entonox had greater plasma beta-ELI concentrations than those who received pethidine. A significant difference in the second stage beta-ELI existed between those who received only Entonox and those who received only pethidine. Induction and augmentation of labour were also associated with differences in beta-ELI. Patients in spontaneous labour had significantly lower beta-ELI than patients who had either artificial rupture of membranes or Syntocinon augmentation of labour. PMID- 6279135 TI - Evaluation of electrical fields inside a biological structure. AB - A digital computer simulation has been carried out of exposure of a cell, modelled as a multilayered spherical structure, to an alternating electrical field. Electrical quantities of possible biological interest can be evaluated everywhere inside the cell. A strong frequency selective behaviour in the range 0 10 MHz has been obtained. PMID- 6279136 TI - Ultrasonic treatment of experimental animal tumours. AB - Studies on the effects of ultrasound on several solid tumours in experimental animals have indicated that tumour growth rates can be reduced. These data are generally consistent with a thermal mechanism of action. Application of combined ultrasound and X-irradiation have shown that with some experimental animal tumours the radiation dose required to locally control 50% of the tumours can be reduced by ultrasound. These results were also consistent with a thermal mechanism of action hypothesis. Fractionated X-irradiation was not enhanced as much as single dose. Pulsing the ultrasound with the same time-average intensity resulted in the same radiosensitivity enhancement. The combined effects of ultrasound and cancer chemotherapy drugs have been studied in mouse leukaemia. The treatment was applied in vitro with cells in suspension. Subsequent to treatment, the cells were inoculated into host mice and survival was monitored. Cytotoxic action of 5 of 10 drugs studied was enhanced by ultrasound. A thermal mechanism of action apparently was not involved. Cavitation in the suspension probably played a role in the cytotoxic enhancement. Experimental data are consistent with the concept that ultrasound causes rapidly reversible cell damage which, in the presence of cytotoxic drugs, is not so readily reversed and results in significant loss of lethal potential of the malignant cells to the host. PMID- 6279137 TI - Basement membrane components and galactosylhydroxylysyl glucosyltransferase in suction blisters of human skin. AB - Basement membrane components and collagen biosynthesis were studied in suction blisters in human skin. The basement membrane components were characterized by immunofluorescence using specific antibodies to type IV collagen, laminin and fibronectin, and collagen biosynthesis was studied by assaying galactosylhydroxylysyl glucosylatransferase. In suction blisters, the separation of epidermis and dermis occurred above the lamina lucida, indicating that the basement membrane, composed of lamina lucida and lamina densa, forms a mechanically strong entity. During the regeneration phase of blisters, type Iv collagen and laminin were not observed in the old epidermal blister roof. This indicates that keratinocytes when separated from the underlying basement membrane or connective tissue do not synthesize laminin or type IV collagen. Galactosylhydroxylysyl glucosyltransferase activity could be demonstrated in blister fluid and was about the same as in serum when expressed on the basis of protein in fresh blisters. It increased by 2-3 fold during the repair of blisters, indicating that there was local production of this enzyme. Further studies revealed that pure epidermis contained galactosylyhdroxylysyl glucosyltransferase and hydroxyprolineand this suggests that epidermis may synthesize some collagen type which, according to these studies, is not type IV (basement memebrane) collagen. PMID- 6279138 TI - Mortality patterns of rock and slag mineral wool production workers: an epidemiological and environmental study. AB - An epidemiological and environmental study of rock and slag mineral wool production workers was undertaken at a plant that has been in operation since the early 1900s. Size characteristics of fibres produced by each process at the plant and data from industrial hygiene surveys were used to evaluate current and past exposures. These data suggest that the average historical airborne fibre concentration probably did not exceed 2.5 fibres/cc before 1935 and 1.0 fibre/cc after 1935. A retrospective cohort mortality study was designed to assess mortality patterns. Detailed occupational histories were compiled on all plant employees. All jobs in the plant were assigned to one of eight potential exposure categories to assess the extent and severity of mineral wool exposure and the effect of other significant exposures on employee mortality. Findings included an increase in the number of deaths due to cancer of the digestive system and non malignant respiratory disease among workers who had over 20 years' exposure to mineral wool or who had survived 20 years since their first exposure to mineral wool. These findings are not inconsistent with those of Enterline's (Symposium on Biological Effects of Mineral Fibres, Lyon, France, September 1979) in the Thermal Insulation Manufacturers' Association's mortality study of men employed in four mineral wool plants. PMID- 6279140 TI - Analysis of fibres in human lung tissue. AB - Standard UICC crocidolite fibres and fibres extracted from occupationally exposed human lung tissue have been analysed by scanninng electron microscopy after different preparation procedures. A significant fibre loss has been shown due to the adhesion of fibres to the breakers. Fibres are also discarded during the washing and extraction of the lung tissue samples. The recovery was found to be 30 to 40% both for UICC standard specimens and for lung tissue samples. PMID- 6279139 TI - Elemental mercury exposure: peripheral neurotoxicity. AB - Nerve conduction tests were performed on the right ulnar nerve of factory workers exposed to elemental mercury vapour. Time integrated urine mercury indices were used to measure the degree of exposure. Workers with prolonged distal latencies had significantly higher urine mercury concentrations when compared with those with normal latencies. Significant correlations between increasing urine mercury concentrations and prolonged motor and sensory distal latencies were established. Elemental mercury can affect both motor and sensory peripheral nerve conduction and the degree of involvement may be related to time-integrated urine mercury concentrations. PMID- 6279141 TI - Antibody activity against herpes simplex virus in human amniotic fluid. PMID- 6279143 TI - The effect of different levels of wheat bran on iron absorption in rats from bread containing similar amounts of phytate. AB - 1. Iron absorption was studied in weanling rats using balance techniques from semi-synthetic diets containing dried white bread (60.5 g dietary fibre/kg. White & Southgate, unpublished results), brown bread (130.2 g dietary fibre/kg) or wholemeal bread (221.2 g dietary fibre/kg) at a level of 300 g/kg and compared with a control group given a diet containing added FeSO4 at a similar Fe level to that for the bread groups. The dried bread contained 6.2-6.4 g phytate/kg. 2. Absorption of Fe was significantly higher (P less than 0.001) in the control group (0.45) than in the White (0.28), brown (0.31) or wholemeal (0.24) groups. 3. A second experiment was carried out on 6-week-old rats in which the dried bread was extrinsically labelled with 59Fe and absorption from a single meal measured by both faecal excretion and incorporation of 59Fe into the blood. Control animals were given 59FeSO4 for comparison. 4. The excretion of 59Fe (% of administered dose) was significantly lower (P less than 0.001) in the control group (31) than in the white (48), brown (45) or wholemeal (47) groups. After 10 d the control group had significantly more 59Fe in the blood than the bread groups, but there were no differences between the bread groups. 5. It appears that wheat bran fibre itself has no effect on the retention of Fe from the diet in the rat, when supplied in amounts similar to those found in commercially available bread. PMID- 6279142 TI - Studies on vitamin D metabolism in malnourished children. AB - 1. Basal levels of serum 25-hydroxy vitamin D (25-OHD) were estimated in ten normal children and twenty-seven children with protein-energy malnutrition. 2. Five normal children and fourteen malnourished children were administered a single massive dose of 15 mg vitamin D orally and the remaining children received oral supplements of 50 micrograms vitamin D daily for 20 d. Blood samples were obtained after 10 and 20 d of dosing and serum 25-OHD levels were repeated. 3. The basal serum 25-OHD levels were significantly lower (P less than 0.05) in malnourished children than in normal children. 4. Administration of a single massive dose of vitamin D as well as daily supplementation of small doses resulted in significant increases in serum 25-OHD levels within 10 d. However, the increase with a massive dose was of a higher magnitude. There was no significant difference between the normal and malnourished children. 5. The binding capacity of serum to 25-OHD was similar in both the groups. 6. These results suggest that vitamin D metabolism is not altered in protein-energy malnutrition. PMID- 6279144 TI - Comparative digestion of timothy (Phleum pratense) fibre by ruminants, equines and rabbits. AB - 1. The abilities of cattle, sheep, goats, equines and rabbits to digest mature timothy (Phleum pratense) hay were compared. Apparent digestibilities were partitioned into true digestibility, metabolic faecal output (MFO) and fibre digestibility. The aid of the study was to determine the relative effects of fermentation site (among groups) and of body-weight (within groups) on the efficiency of digestion. 2. The ruminants were superior to equines, which were in turn superior to rabbits, in digesting fibre-components of the hay. A large individual variation in digestibility was noted only for the equines. Increasing body-weight was associated with higher digestibility in ruminants, but no such trends were seen in the non-ruminants. 3. The MFO expressed as a proportion of dry matter intake gave similar values for all groups (0.085-0.118). As a proportion of available microbial substrate originating from the feed, the values were found to be 0.167 for the ruminant, 0.425 for the equines and 2.13 for the rabbits. The value for the rabbits shows that their lower tract microflora must obtain energy from non-fibre components of the feed. No appreciable digestion of the generated microbes by the host was suggested by the values obtained for the equines. PMID- 6279145 TI - Structure and dynamics of phospholipid membranes: an electron spin resonance study employing biradical probes. AB - The large zero-field splitting of rigid biradicals makes them important candidates for spin probes of phospholipid membranes. Here we develop an electron spin resonance line-shape model for such probes on the basis of the stochastic Liouville equation. Particular emphasis is given to the slow-diffusional regime, characteristic of bilayers in the gel phase. The theory is employed to study the line shapes of bis(verdazyl) biradicals, incorporated into oriented multibilayers of dimyristoylphosphatidylcholine. Computer simulations of the angular-dependent spectra provide the orientational distribution functions and rotational correlation times of the spin probes. They occupy two different sites in bilayer membrane. The orientational distribution of the spin probes is related to the structure of the phospholipid phases. In the L beta' phase the hydrocarbon chains are uniformly tilted by delta = 23 degrees with respect to the bilayer normal. For the P beta' phase we observe a random distribution of tilt angles from delta = 0 degree to delta = 19 degrees, indicating that the chains orient perpendicular to the local (rippled) bilayer surfaces. This structure has not been established previously. In agreement with other studies we find no tilt for the L alpha phase. The order parameters of the hydrocarbon chains increase with decreasing temperature, jumping from S less than or equal to 0.6 to S greater than or equal to 0.8 at the main transition. From the rotational correlation times of the spin probes, intrinsic bilayer viscosities of 0.08 P less than or equal to eta less than or equal to 20 P (50 degrees C greater than or equal to T greater than or equal to 1 degree C) are determined. An Arrhenius plot provides activation energies of the viscous flow. The values increase from Evisc approximately 10 kcal/mol in the L alpha phase to Evisc approximately 18 kcal/mol in the L beta' phase. PMID- 6279146 TI - Binding of 2-azidoadenosine [beta-32P]diphosphate to the receptor on intact human blood platelets which inhibits adenylate cyclase. PMID- 6279147 TI - Resolution of mitochondrial NADH dehydrogenase and isolation of two iron-sulfur proteins. AB - The low molecular weight NADH dehydrogenase which can be solubilized from the mitochondrial NADH-ubiquinone oxidoreductase complex with chaotropic agents consists of three subunits in equimolar ratio [Galante, Y. M., & Hatefi, Y. (1979) Arch. Biochem. Biophys. 192, 559]. The largest subunit (subunit I) can be completely separated from the other two (subunits II + III) by treatment with sodium trichloroacetate and ammonium sulfate fractionation. Both the subunit I and subunit II + III fractions contain iron and acid-labile sulfur. From visible and EPR spectroscopy and the iron and acid-labile sulfide content, we propose that the subunit II + III fraction contains a binuclear cluster. The cluster structure present in subunit I is as yet unclear. On separation of the subunits of NADH dehydrogenase, the FMN is lost. PMID- 6279148 TI - Iron-sulphur cluster composition and redox properties of two ferredoxins from Desulfovibrio desulfuricans Norway strain. AB - Two ferredoxins from Desulfovibrio desulfuricans, Norway Strain, were investigated by EPR spectroscopy. Ferredoxin I appears to be a conventional [4Fe 4S]2+;1+ ferredoxin, with a midpoint reduction potential of -374 mV at pH 8. Ferredoxin II when reduced, at first showed a more complex spectrum, indicating an interaction between two [4Fe-4S] clusters, and probably, has two clusters per protein subunit. Upon reductive titration ferredoxin II changed to give a spectrum in which no intercluster interaction was seen. The midpoint potentials of the native and modified ferredoxin at pH 8 were estimated to be -500 and -440 mV, respectively. PMID- 6279149 TI - Inactivation and solubilization of opiate receptors by phospholipases A2. AB - (1) As previously shown, stereospecific binding of opiates to membrane bound receptors is inhibited by treatment with small amounts of phospholipase A2 from Vipera russelli. This effect is quantified and compared with the enzymes from the venoms of Naja Naja siamensis, Apis Mellifica and from porcine pancreas. All enzymes are equally effective. The inhibition is due to partial phospholipid hydrolysis leading to inactivation of membrane-bound receptor. (2) Bee venom phospholipase A2 together with the synergistically acting peptide, melittin, causes receptor solubilization up to 80% of preformed receptor-ligand complex can be solubilized in this manner. (3) Lysophosphatidylcholine, a product of phospholipid hydrolysis, solubilizes performed receptor-ligand complex to a similar extent. Several other detergents were tested for their ability to solubilize receptor-ligand complex. Digitonin appears to be most effective in solubilizing such a complex. PMID- 6279151 TI - Inhibition by amiloride of sodium transport into rabbit kidney medulla microsomes. AB - Sodium transport into rabbit kidney medullar microsomes was 50% inhibited by amiloride. This Na+ uptake was shown to represent transport when the uptake process was reserved by the ionophore nigericin. The transport was complete within 60 min and proportional to the microsomal protein concentration. The effect of amiloride on transport was specific since the similar compound sulfaguanidine failed to affect microsomal Na+ transport. Amiloride-sensitive Na+ transport into microsomes was inhibited 70% by decreasing the pH (from 7.0 to 5.9), but was unaffected by the presence of a pH gradient. The kinetics of Na+ transport could be explained by a simple model, assuming that amiloride lowered the rate of Na+ entrance into the vesicles but had not effect on the rate of efflux. The failure of amiloride to effect efflux from the vesicles was also demonstrated directly. PMID- 6279150 TI - Ca2+-dependent K+ transport in the Ehrlich ascites tumor cell. AB - The possible presence and properties of the Ca2+-dependent K+ channel have been investigated in the Ehrlich ascites tumor cell. The treatment with ionophore A23187 + CA2+, propranolol or the electron donor system ascorbate-phenazine methosulphate, all of which activate that transport system in the human erythrocyte, produces in the Ehrlich cell a net loss of K+ (balanced by the uptake of Na+) and a stimulation of both the influx and the efflux of 86Rb. These effects were antagonized by quinine, a known inhibitor of the Ca2+-dependent K+ channel in other cell systems, and by the addition of EGTA to the incubation medium. Ouabain did not have an inhibitory effect. These results suggests that the Ehrlich cell possesses a Ca2+-dependent K+ channel whose characteristics are similar to those described in other cell systems. PMID- 6279152 TI - Sodium channel activity of lobster nerve membrane treated with purified phospholipase A2. AB - Sodium channel activity was determined by measuring the veratridine-tetrodotoxin sensitive sodium influx in reconstituted vesicles prepared from lobster nerve membrane and soybean lipids. The sodium channel activity was abolished by treatment of membranes, prior to reconstitution, with purified phospholipase A2. When the treatment with phospholipase A2 was carried out in the presence of bovine serum albumin the channel activity was fully preserved. Electron microscopy revealed that the normal vesicular appearance of the membranes was changed to an amorphous mass by treatment of the membranes with enzyme alone. A population of preserved vesicular structures was observed when bovine serum albumin was present during the enzyme treatment. Analysis of the membrane components indicate that there is no relationship between phospholipid composition and sodium channel activity. PMID- 6279153 TI - Effect of 2H2O/H2O replacement on the dielectric structure of lipid bilayer membranes. AB - Dielectric dispersion measurements were made on planar egg phosphatidylcholine and egg phosphatidylcholine/cholesterol bilayer membranes in H2O and 2H2O electrolyte solutions. The properties of the dielectrically distinct substructural layers of these membranes, which can be characterised by this technique, were found to be insensitive to replacement of H2O by 2H2O. PMID- 6279154 TI - Purification and characterization of an (Na+ + K+)-ATPase proteolipid labeled with a photoaffinity derivative of ouabain. AB - Highly purified lamb kidney (Na+ + K+)-ATPase was photoaffinity labeled with the tritiated 2-nitro-5-azidobenzoyl derivative of ouabain (NAB-ouabain). The labeled (Na+ + K+)-ATPase was mixed with unlabeled carrier enzyme. Two proteolipid (gamma 1 and gamma 2) fractions were then isolated by chromatography on columns of Sepharose CL-6B and Sephadex LH-60. The two fractions were interchangeable when rechromatographed on the LH-60 column, suggesting that gamma 1 is an aggregated form of gamma 2. The total yield was 0.8-1.5 mol of gamma component per mol of catalytic subunit recovered. This indicates that the gamma component is present in stoichiometric amounts in the Na+ + K+)-ATPase. The proteolipids that were labeled with NAB-ouabain copurified with the unlabeled proteolipids. PMID- 6279156 TI - Conditions for sliding of nucleosomes along DNA: SV 40 minichromosomes. AB - 'Sliding' of nucleosomes along DNA under nearly physiological conditions was studied using treatment of SV 40 minichromosomes with the single-cut restriction endonucleases EcoRI and BamHI. Each enzyme can convert no more than 20-25% of the circular DNA molecules of minichromosomes into the linear form irrespective of the presence of histone H1. This suggests absence of the nucleosomes lateral migration (sliding) along DNa at least in the vicinity of the restriction endonucleases cleavage sites during several hours of incubation. The sites available for EcoRI and BamHI in minichromosomes seem to be located predominantly in the spacer DNA regions of nucleosomes. Introduction of only one double-strand (but not single-strand) break into the DNA of minichromosomes stripped of histone H1 is sufficient to induce redistribution of the nucleosome core particles due to their sliding along DNA. Thus, sliding of the nucleosome core particles can be induced under physiological conditions by rather low energy expenditures. PMID- 6279155 TI - The effects of glutaraldehyde and osmium tetroxide on the erythrocyte membrane. A spin label study. AB - A nitroxide spin label probe technique was applied to study the interaction between glutaraldehyde or osmium tetroxide (OsO4) amd the membranes of horse erythrocytes, ghosts and liposomes prepared from erythrocyte lipids. Two major conclusions have been established: (1) Reaction of the fixation reagents with the membrane is selective. OsO4 reacts predominantly with lipids and glutaraldehyde with membrane proteins. (2) The lipid-protein interactions change after pretreatment by OsO4 or glutaraldehyde. PMID- 6279157 TI - Structural homogeneity of mitochondrial DNA in the mitochondrial nucleoid of Physarum polycephalum. AB - Mitochondrial DNA (mtDNA) of Physarum polycephalum was isolated gently by CsCl centrifugation. The mtDNA was linear with molecular weights ranging from 25 . 10(6) to 45 . 10(6) and heterogeneous in size. Nevertheless, thermal transition profiles of the mtDNA suggested that this DNA fraction was more homogeneous than nuclear DNA. Exhaustive digestions of this DNA with restriction endonucleases yielded unique fragments, and then the total of their molecular weights of each digest was around 45 . 10(6). This value is equivalent to the maximum molecular weight estimated using electron microscopy and electrophoresis. Moreover, EcoRI digests of the mtDNA fractionated by the sucrose gradient showed unequimolar quantities of large fragments and a high background between bands. These results suggest that the mtDNA of Physarum has a homogeneous base sequence, and that the size heterogeneity of the mtDNA is attributable to degradation of the DNA under isolation procedures. The mtDNA was cleaved by EcoRI and XhoI to yield 16 and 7 fragments, respectively. A physical map of these fragments was constructed using the routine mapping procedures. The physical map showed that the mitochondrial genome of Physarum was linear with molecular weight of 45 . 10(6). We concluded therefore that the mitochondrial nucleoid is a structure in which the homogeneous mtDNA is highly amplified. PMID- 6279158 TI - Complex kinetics of bis(4-methylumbelliferyl)phosphate and hexadecanoyl(nitrophenyl)phosphorylcholine hydrolysis by purified sphingomyelinase in the presence of Triton X-100. AB - We have examined the hydrolysis of the synthetic phosphodiesters, bis(4 methylumbelliferyl)phosphate and hexadecanoyl(nitrophenyl)phosphorylcholine, by purified placental sphingomyelinase (sphingomyelin cholinephosphohydrolase, EC 3.1.4.12) in the presence of Triton X-100. Triton X-100 enhanced activity with bis(4MU)phosphate at all concentrations tested. At very low concentrations of detergent, bis(4MU)phosphate hydrolysis approached zero. Our results indicate that bis(4MU)phosphate does not form a micelle with Triton X-100. The observed enhancement of bis(4MU)phosphate activity with Triton X-100 is likely due to a direct effect of detergent on the enzyme itself. HDNP-phosphorylcholine formed its own micelle (or liposome) in the absence of Triton X-100 and, at substrate concentrations below 4 mM, hydrolysis was inhibited by Triton X-100. The extent of this inhibition varied with detergent concentrations but could be totally eliminated at substrate values above 4 mM. For theoretical reasons kinetic constants which could be obtained with the HDNP-phosphorylcholine substrate at concentrations above 4 mM are not considered to be truly representative of the real values. We conclude that neither substrate is recommended to describe the true kinetic parameters pertaining to purified sphingomyelinase. In addition, bis(4MU)phosphate may not be suitable as an aid for diagnosis of sphingomyelinase deficiency states.U PMID- 6279159 TI - Chemical, but not functional, differences between the iron-binding sites of rabbit transferrin. PMID- 6279160 TI - Effect on protein stability of reversing the charge on amino groups. AB - The amino groups of beta-lactoglobulins A and B, cytochrome c and ribonuclease were progressively converted to acidic groups by reaction with succinic anhydride. The mixtures of modified proteins generated in this way were analyzed by urea-gradient electrophoresis, which separates the molecules on the basis of their net charge and demonstrates visually their urea-induced unfolding transitions. Molecules succinylated to varying extents were resolved by the electrophoresis, so purification of the many modified species was not required. It is demonstrated that accurate estimates of the stability of the folded state of an individual species may be estimated very easily from its urea-gradient electrophoretic pattern. Changes in ionization of the protein upon unfolding may also be detected. The general electrostatic effect of varying the net charge on these proteins was small. Converting the normally basic ribonuclease and cytochrome c to neutral and then to acidic proteins caused the net stabilities of their folded states to vary by no more than a few kJ/mol. However, specific interactions between a few ionized groups appear to be more important in some instances. Succinylation of the 19th, and final, lysine residue of cytochrome c produced unfolding even in the absence of urea, whereas reaction of the first 18 had very little effect. Reaction of the initial amino groups of beta lactoglobulins A and B produced a small increase in stability in a few instances, a decrease in others; modification of more than about ten groups abruptly caused unfolding in the absence of urea. PMID- 6279161 TI - Identification of plasma kallikrein as an activator of latent collagenase in rheumatoid synovial fluid. AB - Rheumatoid synovial fluid contains an activator of latent collagenase from culture medium of pig synovium. The activator was purified by gel chromatography on Ultrogel AcA 44 and affinity chromatography on soybean trypsin inhibitor coupled to Sepharose 4B. The purified material was homogeneous on SDS polyacrylamide gel electrophoresis with Mr 88 000. The activator had limited proteolytic activity against azo-casein, but showed amidase activity on Pro-Phe Arg-NMec, Z-Phe-Arg-NMec, D-Val-Leu-Arg-NPhNO2 and D-Pro-Phe-Arg-NPhNO2, with an optimum at pH 8.0. Activity was completely inhibited by diisopropyl fluorophosphate, soybean trypsin inhibitor, leupeptin and Pro-Phe-Arg-CH2Cl, whereas lima bean trypsin inhibitor, Tos-Lys-CH2Cl, a specific inhibitor of factor XIIa from maize, EDTA and iodoacetate were not inhibitory. These properties of the activator suggested that it might be plasma kallikrein (EC 3.4.21.34), and the possibility was further examined. The activator was treated with [3H]diisopropyl fluorophosphate, and run in SDS-polyacrylamide gel electrophoresis with reduction; a radioautograph of the gel showed a pair of [3H]diisopropyl phosphoryl-labelled bands (Mr 36 000 and 34 000) identical to those obtained with authentic plasma kallikrein. Double immunodiffusion with monospecific antiserum against human plasma kallikrein confirmed the identification. This is the first demonstration of collagenase-activating activity of plasma kallikrein, and raises the possibility that activation of prokallikrein in the inflamed joint space may contribute to the disease process not only by the production of bradykinin, but also by activating latent collagenase. PMID- 6279162 TI - Activation of calmodulin-dependent NAD+ kinase by trypsin. AB - Sea urchin egg NAD+ kinase (ATP:NAD+ 2'-phosphotransferase, EC 2.7.1.23), a calmodulin-dependent enzyme, can be activated by a moderate treatment with trypsin in a similar fashion to calmodulin. Stimulation by trypsin is dependent on its concentration (half-maximal dose: 1.5 microgram/ml) but independent of the presence of calcium. This suggests that limited proteolysis is able to activate NAD+ kinase as described for several other calmodulin-activated enzymes and that these enzymes may interact with calmodulin in a similar way. PMID- 6279163 TI - Electron paramagnetic resonance studies of NO-heme-nitrogen base. An interpretation of electron paramagnetic resonance spectra of NO-hemoproteins. AB - In order to characterize the structure of the heme environment of hemoproteins, electron paramagnetic resonance (EPR) spectra for the NO complex of the iron(II) porphyrin nitrogen bases (pyridine and imidazole derivatives) were measured. The coupling constants of the nitrogen atom of NO (AN1) and the fifth ligand (AN2) and g values were determined from the 9-lined hyperfine using second-derivative display. These EPR parameters varied with changes in trans ligand (trans effect) and heme substitution at positions 2 and 4 (cis effect) as follows. (1) Both AN1 and AN2 increased as the basicity of the nitrogen atom of the fifth ligand increased, while AN1 increased concomitant with the decrease of AN2 by steric hindrance of the fifth ligand. (2) Both AN1 and AN2 increased as the basicity of the porphyrin nitrogen atom decreased. (3) In both cases, the anisotropy of g values (gx and gy) decreased concomitant with the increase of AN2. From the analysis of the EPR spectra of model systems, the substantial difference in the EPR spectra of NO-hemoproteins is discussed in relation to iron-proximal histidine binding and heme-apoprotein interactions. PMID- 6279164 TI - Analysis of strain-induced EPR-line shapes and anisotropic spin-lattice relaxation in a [2Fe-2S] ferredoxin. AB - The electron paramagnetic resonance spectrum of the [2Fe-2S]1+(2+;1+) cluster in spinach-leaf ferredoxin has been measured at four microwave frequencies from 1 to 35 GHz. Using a modified g-strain formula, the asymmetrical spectrum has been simulated in detail without the assumption of signal multiplicity. In all but the lowest frequency bands the line width is dominated by an extremely anisotropic g shift distribution, caused by a statistical distribution in dislocation strains. The crossover point of domination by unresolved proton splittings is around 2 GHz. The angle-dependent elasticity of the cluster can be related to an anisotropy in the spin-lattice relaxation rate. Intensity behaviour under continuous saturation, at temperatures in the two-phonon region, is in qualitative agreement with elementary theory. On the basis of these results it is argued that biochemists should be aware of the questionable nature of some ad hoc assumptions commonly made to interpret EPR of metalloproteins. Specifically, a physically meaningful determination of the number and stoicheiometry of distinguishable compounds, represented in a complex spectrum, may well require more advanced theoretical tools than the frequently employed deconvolution in symmetrical Gaussians with associated unique relaxation times. PMID- 6279165 TI - Glycerol kinase activity and glycerol metabolism of rat granular pneumocytes in primary culture. AB - Glycerol kinase activity and glycerol utilization by rat granular pneumocytes were determined in order to investigate the rate-limiting step for glycerol incorporation into lung lipids. Granular pneumocytes were isolated in primary culture following trypsinization of rat lungs. Glycerol kinase activity was 8.2 nmol/h per 10(6) cells. Incorporation of [1,3-14C]glycerol into total cell lipids was 0.29 nmol/h per 10(6) cells. In the presence of saturating glycerol concentration, production of 3H2O from [2-3H]glycerol was 13 times greater than incorporation of [14C]glycerol into lipids. Glycerol phosphate dehydrogenase activity in isolated cells was approximately 10 times glycerol kinase activity. In the presence of 5.6 mM glucose, glycerol incorporation into lipids was decreased 79% and detritiation of glycerol was decreased 34%. This effect of glucose was due to a 25% increase in cell glycerol 3-phosphate content, resulting in dilution of the precursor pool and possible inhibition of glycerol phosphorylation. These results indicate that the relatively limited incorporation of glycerol into surfactant phospholipids by lung epithelial cells reflects the relatively high rate of glycerol 3-phosphate oxidation. PMID- 6279166 TI - Elevated levels of cellular and extracellular phospholipases from pathogenic Naegleria fowleri. AB - Phospholipase A, sphingomyelinase and lysophospholipase activities were examined in cell homogenates and cell-free culture media of virulent and virulent attenuated Naegleria fowleri and nonpathogenic Naegleria gruberi. Homogenates of virulent N. fowleri contained from 3 to 250 times the lipolytic activity of virulent-attenuated and non-pathogenic Naegleria spp. Similarly, the cell-free media of virulent N. fowleri cultures contained large quantities of phospholipase A, lysophospholipase and sphingomyelinase while comparable activities in the cell free media of virulent-attenuated and nonpathogenic Naegleria spp. were only slightly, if at all, detectable. Lipolytic enzymes accumulated in the media of virulent N. fowleri cultures at various stages during growth but not in virulent attenuated and nonpathogenic Naegleria cultures. In general, phospholipase A and sphingomyelinase accumulated during the log phase of growth while lysophospholipase appeared only in the late stationary phase. We conclude that pathogenic Naegleria contain potent lipolytic enzymes that are released selectively into the media during growth. These enzymes could contribute to the pathogenesis of Naegleria-induced primary amoebic meningoencephalitis. PMID- 6279167 TI - Linkage of the isoprenoid biosynthetic pathway with induction of DNA synthesis in mouse lymphocytes. Effects of compactin on mitogen-induced lymphocytes in serum free medium. AB - Concanavalin A induction of DNA synthesis in mouse spleen lymphocytes cultured in serum-free medium was shown to be very sensitive to inhibition by compactin (ML 236B), a specific competitive inhibitor of hydroxymethylglutaryl-CoA reductase. As low as 0.1 microM compactin could give 98% inhibition of mitogen induction of a 5.10(6) cells/ml culture. This inhibition could be reversed completely by addition of exogenous mevalonate, but could not be reversed by either exogenous cholesterol or isopentenyladenine. Oxygenated sterol inhibition of mitogen induced DNA synthesis could be reversed by cholesterol or by mevalonate, whereas cyclic AMP inhibition could not be reversed by either compound. These results suggest that endogenous cholesterol production is a necessary but not sufficient factor co-ordinated with mitogen-induced DNA synthesis, and that the presence of some additional product of mevalonate metabolism is involved also. Isopentenyladenine, though, did not have as significant effect of alleviating any of the above inhibitions. Since mevalonate could not relieve cyclic AMP inhibition, but could overcome compactin inhibition, cyclic AMP inhibition cannot be explained as due only to blockage of mevalonate production. PMID- 6279168 TI - [ESR study of interaction between adenylate kinase, substrates and Mn2+ ions]. AB - Plant adenylate kinase was first investigated by ESR technique at room temperature. In contrast to previous studies the measurement conditions corresponded to maximal enzyme activity. It was shown that adenylate kinase addition provoked Mn2+--adenine nucleotide binary complex distribution and rearrangement of components with the formation of ternary Mn2+--adenine nucleotide--adenylate kinase complex. The same results were obtained in the studies on myokinase from rabbit muscle. PMID- 6279170 TI - [Selectivity characteristics of cation channels in Nitella flexilis plasmalemma]. AB - Ionic selectivity of Nitella flexilis plasmalemma cation channels is studied by voltage-clamp method with consecutive replacing of cations in the bathing medium. The selectivity sequence received by measuring the ionic current reversal potentials, psi alpha is: Ba++ approximately equal to Sr++ approximately equal to Ca++ greater than Mg++ greater than Cs+ approximately equal to K+ greater than Na+ greater than Li+. An analysis of results based on the three-barrier channel model suggests that when ions of the same valency are compared, the channel selectivity is determined by specific interactions between the ion and the nearest water molecules, which is possible both in a narrow and wide pore. On the other hand, when monovalent and divalent ions are compared the effects of ions binding in the channel or near the membrane surface prevail, thus causing the channel preference for divalent cations. PMID- 6279169 TI - [Comparative study of beta-adrenoreceptors of rat brain synaptic membranes]. AB - Affinity of beta-adrenoreceptors in the rat brain synaptic membranes to agonists isoproterenol and norepinephrine, as well as to antagonist 125I hydroxybenzylpindolol is lower in young (1 month) and old (24--26 months) than in mature (8--12 months) rats. Desensitization toward isoproterenol is expressed in the young ones only. In the old but not in other groups simultaneous action of isoproterenol and N-ethylmaleimide decreases the following binding of the antagonist while the same agents added separately produced no effect. It is suggested that beta-adrenoreceptors undergo age-related changes in their conformational state due to modification of the membrane environment. PMID- 6279171 TI - [Channel formation properties of black widow venom]. AB - Black widow venom in the concentration 1--10 mkg/ml added on one side of the bilayer of common bovine brain phospholipids induces the formation of conductivity channels with high cation-anion selectivity with the number of cation transfer for K+, Ca2+, Sr2+, Mg2+, Na+, Cs+, Li+ equaling 0.98, 0.97, 0.96, 0.94. 0.88. 0.82, 0.82 correspondingly (at pH = 7.5). At pH less than 3.5 the channels are slightly selective for anions (the number of cation transfer 0.4). Potential-dependence of the channels is found, which is explained by microstructural reconstruction of their protein complex. PMID- 6279172 TI - Microsomal cAMP-independent histone H1 kinase activity in plasmacytoma, Morris hepatoma and normal liver. AB - A protein kinase activity with high specificity for histone H1 was isolated from mouse plasmacytoma, Morris hepatoma and normal mouse liver and compared by ion exchange chromatography after DEAE-cellulose, hydroxylapatite and Sephadex G-200 chromatography. This cAMP-independent histone H1 kinase is not affected by the heat-stable cAMP-dependent protein kinase inhibitor. It has the following particular properties: it prefers GTP to ATP as substrate and was found to be present with a great activity only in neoplastic tissues. No phosphatase activity was detected in the partially purified histone H1 kinase fraction from normal and neoplastic cells. These results suggest either an increase amount of histone H1 kinase and/or of its activator in neoplastic cells, or the presence of a strong inhibitor in normal cells. This histone H1 kinase appears to be analogous to the chromatin bound kinase which phosphorylates histone H1 at the NH2 and COOH terminal regions. We might suggest an implication of this kinase in the regulation of cell division. PMID- 6279173 TI - [Role of membrane colchicine binding proteins in the transmission of prolactin message to casein genes in the rabbit mammary gland]. AB - Previous work demonstrated that tubulin binding drugs specifically inhibit the capacity of prolactin to initiate casein and DNA synthesis in the mammary cell. It was concluded that microtubules or other tubulin containing cellular structures were involved in the transmission of the prolactin message to genes. In the present work, it is shown that griseofulvin, an antimitotic drug which alters microtubule structure and function, does not prevent prolactin actions. Autoradiographic studies showed that [3H]colchicine binds preferentially to plasma and Golgi membranes in the mammary cell. Short term cultures of mammary explants with [3H]colchicine demonstrated that the labelled drug binds to membranous cellular structures which were isolated from explants at the end of the culture. Fractions containing plasma and Golgi membranes contained the highest amount of radioactivity. Solubilisation of the membranes by Triton X-100 dissociated the [3H]colchicine from the prolactin receptors as judged by a chromatography of the soluble fraction on a Sepharose 6 B column. On the column, the labelled colchicine remains associated with a molecular entity which may be free tubulin. In all cases, the binding of [3H]colchicine was greatly attenuated by an excess of unlabelled colchicine but was only slightly affected by the competition with lumicolchicine. These results suggest that mammary membranes contain tubulin and that binding of drugs to this molecule inhibits the generation of the prolactin second messengers eliciting the hormonal actions in the mammary cell. This also suggests that microtubules are probably not involved in the mechanism of prolactin action. PMID- 6279174 TI - [Inventory of proteolytic activity of a new collagenolytic bacteria Empedobacter collagenolyticum]. AB - During the growth of Empedobacter collagenolyticum on a medium with gelatin, only one proteinase, a collagenase, was excreted in the culture medium. No other proteolytic activity was detected in the extracellular medium or in acellular extracts. The other proteases of this bacteria are principally intracellular peptidases. By electrophoresis of an acellular extract five peptidases were separated; they were aminopeptidases and dipeptidases. Some of them exhibited a specificity towards peptides with aminoacid frequently found in collagen; Gly Leu, Gly-Pro, Pro-Gly-Gly. Two other peptidases seem to have special specificity, one of them hydrolyses peptides with lysine residues at the NH2 terminal end, the other one hydrolyses dipeptides of the structure Lys-X. These enzymes were also found in the culture medium; they certainly play an important role in bacterial nutrition. PMID- 6279175 TI - [Purification and study of some properties of a collagenase produced by Empedobacter collagenolyticum]. AB - A collagenase from Empedobacter collagenolyticum was extracted from the culture medium of the bacteria. The complete purification of the enzyme was achieved by successive ammonium sulfate precipitation. Sephadex G 200 gel filtration and DEAE cellulose chromatography. This collagenase is active on insoluble collagen, and on the synthetic peptide Pz-Pro-Leu-Gly-Pro-D-Arg. Its optimum activity was at 30 degrees C and at pH 7.6. A strong inhibition was observed with chelating agents such as O-phenanthroline and EDTA. Among the cations tested to restore the activity, only Ca2+ has a measurable effect. Heavy metals, Pb, Hg, Cd, Cu, Fe, Co, strongly inhibit the enzyme activity. Zn2+ is also highly inhibitory; 10 microM ZnCl2 completely inhibits the collagenase. p CMB, iodoacetate have little effect on the collagenase. This new collagenase ressembles by most of its properties the already known bacterial collagenases. PMID- 6279176 TI - [A new type of intracellular relay for protein hormones?]. PMID- 6279177 TI - Evidence that the activation of casein gene expression in the rabbit mammary gland can be elicited by a low amount of prolactin firmly retained on its receptors. PMID- 6279178 TI - [Comparison of various physico-chemical properties of pig and cattle myeloperoxidases]. AB - The myeloperoxidases possessing the specific activity of 500 000-700 000 of o dianisidine units per 1 mg of protein were isolated and purified from the lysosome-like granules of pig and cattle neutrophylic leukocytes. The absorbance spectra for the enzymes in the visible region are identical; their molecular weight is 140 000-160 000. The enzymes are made up of two subunits, each containing a heavy (m. w. 68 000) and a light (m. w. 10 000) polypeptide chains. The pH optimum for myeloperoxidase oxidation of o-dianisidine lies around 5.8-6.2 for both enzymes. The dependence of the reaction rate on H2O2 concentration does not obey the Michaelis--Menten kinetics. The optimal concentrations of the reaction substrates are 0.34 mM for o-dianisidine and 0.075 mM for H2O2. The amino acid composition and the peptide maps for pig and cattle myeloperoxidases are in many ways similar, showing no coincidence of enzymes, however, in terms of antigenic determinants. The similarities of some physico-chemical properties of pig and cattle enzymes and their immunological differences are discussed. PMID- 6279179 TI - [Effect of cardenolids and sodium ion gradient on ATP-dependent Ca2+ accumulation in cardiac sarcolemmal vesicles]. AB - Sarcolemmal preparations isolated from rat and guinea pig hearts mostly consisting of inside-out membrane vesicles catalyzed ATP-dependent Ca2+ accumulation. The creatine kinase ATP-regenerating system containing exogenous creatine kinase and phosphocreatine was most effective in supporting Ca2+ accumulation. The normalized rate of Ca2+ accumulation obtained by dividing the Ca2+ uptake rate by its initial equilibrium vesicular content was correlated with the (Na+ K+)-ATPase activity. The ATP-dependent Ca2+ uptake in sarcolemmal vesicles was inhibited by cardenolids--digitoxigenin and ouabain (40-50%), when the latter acted from inside the vesicles. The Ca2+ gradient formed in sarcolemmal vesicles at the expense of ATP was dissipated by Na+ but not by Li+ added into the external medium. The external sodium ions also caused Ca2+ efflux from sarcolemmal vesicles equilibrated with Ca2+. The described effects of Na+ are taken to show the existence of a Na-Ca exchange system in cardiac sarcolemma. Ca2+-ATPase of sarcoplasmic reticulum (SR) vesicles from guinea pig heart as well as ATP-dependent Ca2+ influx in these preparations were found to be partially suppressed by digitoxigenin. External NaCl led to be a rapid (during 5-10 sec) fall in vesicular Ca2+ content (by about 40%) accumulated at the expense of ATP followed by a return of Ca2+ content to its initial level. LiCl had no effect on Ca2+ content in SR. NaCl gradient directed inside the SR vesicles did not influence the distribution of Ca2+ between internal and external vesicular volumes in equilibrium (without ATP). The differences in properties of the Ca pump of sarcolemma and sarcoplasmic reticulum found in this work support the idea on the existence of sarcolemmal system of ATP-dependent transport of Ca2+. PMID- 6279180 TI - [Endogenous phosphorylation of sarcoplasmic reticulum fragments of rabbit fast skeletal muscles]. AB - The purified membrane fragments of sarcoplasmic reticulum (SR) of rabbit fast skeletal muscles were found to incorporate 32P from[gamma-32P]ATP in endogenous membrane substrates and in histone H1. The existence of membrane-bound protein kinase of SR was demonstrated by steady state binding of [3H]-cAMP to the SR membranes. The constant of [3H]cAMP binding to the membranes is 2.5 +/- 0.003 x 10(6) M-1, the number of binding sites is 6.1 +/- 0.8 pmol per 1 mg of protein. The endogenous phosphorylation of SR components was inhibited by cAMP and cGMP at concentrations of 10(-7)-10(-6) and depended on Mg2+ and Ca2+. The thermostable protein inhibitor of cAMP-dependent protein kinase inhibited the endogenous phosphorylation of SR membranes by 30-40%. The protein phosphoproduct of SR membranes revealed the properties of a phosphoester. The membrane-bound protein kinase was active towards the exogenous substrate--histone H1. Phosphorylation in the presence of histones was independent of cyclic nucleotides, Mg2+ and Ca2+. Fractionation of 32P-labelled solubilized membranes in polyacrylamide gel in the presence of Na-SDS showed that the radioactivity is bound to protein zones with molecular weights of 95 000 and 6000. PMID- 6279181 TI - [Immunological detection of sphingomyelin exchange protein in tumors and in embryonic rat liver]. AB - The interaction of postmicrosomal supernatants from the Jensen sarcoma, sarcoma 45, nephroma RA, normal, regeherating and fetal rat livers with antisera to the sphingomyelin exchange protein isolated from hepatoma 27 was studied by double immunodiffusion. It was shown that this protein is present in comparable amounts in all the tumours tested and in fetal rat liver, whereas in normal and regenerating liver its content is negligible. Based on the correlation between the content of sphingomyelin exchange protein in cell cytosols and the amount of sphingomyelin in the corresponding mitochondria it is suggested that this protein is responsible for the presence of sphingomyelin in mitochondria. PMID- 6279183 TI - [Existence of boundary lipids in reconstituted hydrophobic protein-lipid system]. AB - A fraction of hydrophobic proteins (9, 14, 18 kD) soluble in a chloroform methanol mixture (2:1) has been isolated from Micrococcus lysodeikticus bacterial membranes. The proteins obtained were introduced into proteoliposomes at a protein/lipid weight ratio ranging from 0.1 to 0.25 in combination with the fluorescent probe pyrene or the spin probe 2-(14-carboxytetradecyl)-2-ethyl-4.4 dimethyl-3-oxasolidinyloxyl. The excimertization of pyrene upon direct excitation of its molecules (gamma excit.=338nm) and under conditions of energy transfer from the excited protein chromophores to pyrene (gamma excit.=286nm) and the spin spin exchange between the spin probe molecules was investigated. The experimental results suggest that the hydrophobic protein molecules are surrounded by a structurally heterogenous lipid area containing up to 3.3 mg of lipid per l mg of protein. The maximal expression of structural heterogeneity was observed at the minimal content of protein in the proteoliposomes. Treatment with the membranotropic antibiotic gramicidin S resulted in disappearance of lateral heterogeneity of lipids in the constituted system and in lipid aggregation in bacterial membranes. It is assumed that the aggregability of membrane proteins depends on the structural rearrangement of some part of lipid bilayer around them. PMID- 6279182 TI - [Effect of nucleoside triphosphates on cyclic nucleotide phosphodiesterase: role of protein modulators]. AB - The effects of nucleoside triphosphates (ATP and GTP) on phosphodiesterase (PDE) of brain and outer segments of the retina enriched or devoid of protein modulators were studied. In the case of retinal outer segment PDE the enzyme activity was considerably inhibited by both nucleosides only when the enzyme was separated from the inhibitor. In case of brain PDE, on the contrary, the effect of the nucleosides was much more pronounced in the enzyme preparation coupled with the protein activator, calmodulin. The latter when added to brain PDE devoid of the activator in the presence of ATP and GTP considerably reduced the enzyme activity. An addition of the inhibitor simultaneously with GTP to the purified PDE of outer segments increased the PDE activity. The constants for the inhibition of brain PDE coupled with calmodulin and retinal outer segment PDE separated from the inhibitor by ATP and GTP were determined. PMID- 6279184 TI - [Biochemical mechanism of the radioprotective action of adenylic compounds]. PMID- 6279185 TI - Ontogeny of cyclic nucleotides in embryonic chick gonads. AB - The cyclic nucleotides AMP and GMP show characteristic patterns of concentration throughout development in the gonads of the embryonic chick. In the ovary, cyclic AMP concentration varies depending upon whether the ovary is regressing (right) or developing (left). In the regressing right ovary cyclic AMP concentration is 4.25 pmol/10(6) cells on day 8 and 1.3 pmol/10(6) cells on day 15 of embryonic development. In the developing left ovary cyclic AMP is approximately one-half that of the right ovary and also decreases throughout this period (2.9 pmol/10(6) cells on day 8 and 0.75 pmol/10(6) cells on day 12 through hatching). Cyclic GMP levels, on the other hand, are similar in right and left ovaries on day 10 (4 and 5 fmol/10(6) cells, respectively). On day 12, however, cyclic GMP in the regressing right ovary increases 5.5-fold while that in the developing left ovary remains constant. In the testis, cyclic AMP and cyclic GMP remain at 0.9 pmol and 6 fmol per 10(6) cells, respectively, throughout embryonic development. Micromolar concentrations of cyclic AMP and cyclic GMP reciprocally inhibit the hydrolysis of each other to a similar degree in the developing left ovary and testis. In the regressing right ovary, however, the hydrolysis of cyclic GMP is not affected by cyclic AMP. PMID- 6279186 TI - Relaxin receptors in the myometrium and cervix of the pig. PMID- 6279187 TI - Placenta as a source of 'brain' and 'pituitary' hormones. AB - Placental peptides, such as human chorionic gonadotropin (hCG) and human placental lactogen (hPL), which have marked homologies to pituitary peptides, were described in the early part of this century. Recently, the presence in placenta of additional peptides previously demonstrated as occurring in other tissues, such as brain and pituitary, has been reported. Their presence in placenta has been attributed to similar embryological origin of the tissues which share these peptides, although this has by no means been proven. Placental concentrations of these recently described peptides are several orders of magnitude lower than described for their original sites of production. In many instances, definitive characterization of structural identity with their extraplacental counterparts has not been performed, but has been based on indirect evidence obtained by immunoassay or immunocytochemistry. Evidence of placental synthesis has been obtained for hCG, hPL, and pro-opiomelanocortin (the precursor molecule for adrenocorticotropic hormone (ACTH), B-lipotropin (B-LPH), alpha-melanocyte-stimulating hormone (alpha-MSH), beta-endorphin, and other thus far uncharacterized peptides). Possible functions for the recently described peptides might include actions on the maternal or fetal systems or local (paracrine) actins affecting other placental constituents, although to date no definitive physiological roles have been demonstrated. PMID- 6279188 TI - Expression and structures of human placental hormone genes as a function of placental development. PMID- 6279189 TI - Cyclic nucleotides, DNA, and steroid levels in ovarian follicles and corpora lutea of the cyclic hamster. PMID- 6279191 TI - Suppression of luteinizing hormone and testosterone secretion in bulls following adrenocorticotropin hormone treatment. AB - The present investigation was conducted to evaluate the inhibitory effects of adrenal corticosteroids on testosterone production by the bull testis. Administration of a single i.v. dose of adrenocorticotropic hormone (ACTH; 80 IU) resulted in a corticosteroid peak which lasted approximately 6 h. During this 6 h period, no episodic increases in secretion of LH or testosterone were initiated and basal concentrations of testosterone were suppressed (P less than 0.05) below control values. Episodic secretion of LH and testosterone resumed 6--7 h after ACTH when concentrations of serum corticosteroids had returned to basal levels. These results suggest that ACTH-induced increases in serum corticosteroids suppress the episodic secretion of LH, resulting in a suppression of testosterone secretion by the bull testis. PMID- 6279190 TI - Insulin augmentation of testosterone production in a primary culture of rat testicular cells. AB - The direct effects of insulin on basal and human chorionic gonadotropin (hCG) stimulated accumulation of testosterone were investigated in vitro using a primary culture system of rat testicular cells from adult hypophysectomized male rats. The basal accumulation of testosterone was low throughout the 10-day incubation period. Treatment of testicular cells with insulin (10 micrograms/ml) by itself was without effect on the basal accumulation of testosterone, while treatment with increasing concentrations (0.1--10 ng/ml) of hCG resulted in dose dependent increases in the accumulation of testosterone. Furthermore, concomitant treatment with increasing concentrations (0.01--10 micrograms/ml) of insulin led to a dose-dependent augmentation (up to 116% on Day 10) in the hCG-stimulated accumulation of testosterone, as well as a 1.6-fold increase in the testicular responsiveness to hCG. In contrast, treatment with desoctapeptide insulin (10 micrograms/ml), a trypsin degraded insulin, was without effect on the hCG stimulated accumulation of testosterone. Increasing duration (12--72 h) of treatment with insulin resulted in time-dependent increases in the hCG-stimulated accumulation of testosterone achieving statistical significance (P less than 0.05) by 36 h. In addition, pretreatment with insulin (10 micrograms/ml) brought about significant (P less than 0.01) increases in the choleragen and Bt2cAMP stimulated accumulation of testosterone. The augmenting effect of insulin was equally effective upon culturing in a glucose-free medium and was not associated with significant alterations in testicular cell number or cellular DNA or protein content. It is concluded that diminished testicular steroidogenesis in the diabetic rats may represent, at least in part, a direct consequence of insulin deficiency at the testicular level and that insulin may play an important role in the augmentation of testicular androgen production. PMID- 6279192 TI - Biodegradation studies on periodate oxidized cellulose. PMID- 6279193 TI - Prospective study of Epstein Barr virus (EBV) infection during pregnancy. AB - The serological EBV profile of 2752 pregnant women characterized in most cases a latent EBV carrier state. The pregnancy rarely reactived this latent infection. But mothers with an "active" EBV serology gave birth more frequently than others to still born or defective bodies. In six pregnancies with primary EBV infection, detected early, four presented a pathological delivery. Furthermore the follow up of the EBV profile in 719 mothers revealed a significant relation between defective births and persistent EA antibodies. The role of EBV has yet to be precisely defined but the early detection of anti-EA EBV antibodies should be considered as a risk indicator in the management of pregnancy. PMID- 6279194 TI - Hormonal changes in patients with haematological malignancies. AB - Levels of immunologically reactive insulin (IRI), growth hormone (GH), thyroxine, cortisol and ACTH in sera of patients with various haematological malignancies were determined. IRI and GH levels were increased in 80% of the patients regardless of the type of disease. IRI was more elevated in relapse than in remission. Cytostatic treatment returned IRI serum levels to normal. Thyroxine, ACTH and cortisol in serum were higher in only two percent of the patient with haematological malignancies. PMID- 6279195 TI - Aging of the erythrocyte XIII. Decrease in nitroxide reducing capacity. AB - A method is proposed for the determination of nitroxide-reducing capacity (NRC) of erythrocytes, and other cells, based on measurements of reduction of a nitroxide stable free radical in cell suspensions by ESR spectroscopy. Comparison of different age fractions of bovine erythrocytes by this method demonstrates a decrease in NRC during cell aging in vivo. PMID- 6279196 TI - The respective advantages of complement fixation and ELISA for routine diagnosis of cytomegalovirus infection. AB - The results of CF and ELISA tests for cytomegalovirus performed on 270 sera of hospitalized patients show a positive correlation. As a general rule, ELISA is more sensitive than CF, except for a few sera collected from patients with immunological disorders. When two sequential sera are available, the CF remains a reliable and inexpensive method. But when only one serum can be obtained, the probability of an active CMV infection can be estimated on the IgM/IgG ratio. In 26% of the patients, this ratio was greater than or equal to 1. The ELISA is twice as expensive as the CF test. To reduce its cost, a simple method for preparing ELISA antigen from commercially-obtained CF antigens is described. PMID- 6279197 TI - Effects of amphotericin B on membrane permeability--kinetics of spin probe reduction. AB - The effect of the polyene antibiotic amphotericin B on the permeability of both unilamellar and multilamellar model membranes is investigated. The method measures the loss of the electron paramagnetic resonance signal of a spin probe, trapped in the aqueous compartment of a lipid dispersion, upon addition of ascorbate ions to the bulk aqueous phase. Amphotericin B causes large increases in the permeability of cholesterol-containing egg phosphatidylcholine membranes, whereas the effects are small in the absence of sterol and do not depend on surface charge. The effect of amphotericin depends upon the antibiotic:sterol mole ratio. The antibiotic appears to be unable to cross the membrane, acting only on the outermost bilayer of a multibilayer dispersion. When a phospholipid in the gel phase is used, amphotericin B causes large increases in permeability, independently of the presence or absence of sterol. It is suggested that the mechanism of action of amphotericin B is different for lipids in the liquid crystalline or gel states. PMID- 6279199 TI - H-NMR of U-G-A and U-G-A-A in D2O: assignment of nonexchangeable protons and analysis of solution conformation. PMID- 6279198 TI - Proton spin-lattice relaxation rates in erythrocytes adsorbed with hemagglutinating viruses. PMID- 6279200 TI - H-NMR of G-U-C and G-U-C-C in D2O: assignment of nonexchangeable protons and analysis of solution conformation. PMID- 6279201 TI - The chiroptical properties of proteins. III. Adenylate kinase. PMID- 6279202 TI - Autoregulation of repressor synthesis in bacteriophage lambda imm434. AB - A 550-bp DNA fragment derived from the immunity region of phage lambda imm434 and carrying the wild-type prm promoter has been inserted upstream of the lacZ gene, in the beta 2 region of an 'in-vitro'-constructed transducing phage. In strains lysogenized with this phage, the rates of transcription from the 434-prm promoter can be assayed as units of beta-galactosidase in the absence or in the presence of increasing levels of 434 repressor. The results obtained indicate the existence of a mechanism of autoregulation for repressor maintenance in strains lysogenic for wild-type phage 434. The rate of transcription from prm in a (434) lysogen is of the same magnitude as that in a (lambda ) lysogen, and, as in the case of lambda , high levels of repressor repress prm whereas low levels stimulate it. An estimation of the strength of the leftward promoters located in the right half of the lambda beta 2 region compared with 434 prm was also obtained. PMID- 6279203 TI - Cell-membrane receptors for purines. Review. AB - Purines are involved in many aspects of cell chemistry - intermediary metabolism, nucleic acid synthesis, and the supply of high-energy phosphates to various active transport systems. In addition, however, there appear to be specific receptor molecules located within the plasma membrane of some cells, which mediate changes of cell function in response to purines present in the extracellular fluid. It is the purpose of this review to summarize the kind of functions subserved by those receptors as well as the basic structural requirements for their activation. PMID- 6279204 TI - Epidermolysis bullosa. AB - The disorders known as epidermolysis bullosa constitute a group of genetic diseases of the skin which in the mildest form are debilitating throughout life and in the most severe forms result in death at an early age. The profound morbidity associated with all forms of the disease results not only in the loss of an economically productive life but also, in many cases, in the need for continuing medical care throughout life. Inroads are being made into potential pathogenic mechanisms in certain forms of EB. The current state of knowledge can best be characterized as a number of potentially significant disparate observations from various disciplines. The ultimate goals of research in EB are not only to understand the disease but also to provide the biochemical tools for reliable genetic classification, for antenatal diagnosis, and for rational modes of therapy. PMID- 6279205 TI - The biology of pigmentation. AB - The many factors involved in the normal pigmentation of human skin are highly complex involving anatomic, biochemical, and genetic aspects of melanocytes in the skin and the influence of UV light and various hormones on the melanocytes. It is probably more than just coincidence that the melanocytes, which are of neurogenic origin, are so responsive to several trophic hormones produced in the brain (pituitary). Understanding of the various factors involved in the normal pigmentary process is crucial to explaining the many alterations and anomalies in human pigmentation. PMID- 6279206 TI - [Relationship between the kininase and angiotensin-converting lung functions in rabbits with cerebroischemic and renovascular hypertension]. PMID- 6279207 TI - [Penetration of 3H-cyclic adenosine-3',5'-monophosphate into different tissues of mice]. PMID- 6279208 TI - [Regulation of acetylcholinesterase level in microsomes of neurons by acetylcholine and cyclic nucleotides]. AB - In a cell-free system containing isolated nuclei and microsomal-cytoplasmic brain fraction there takes place a spontaneous puromycin-sensitive increase in acetylcholinesterase (AChE) activity of microsomes. As a result of preincubating the system with acetylcholine (ACh) (10(-6)-10(-3) M), AChE activity of microsomes was found to be increased, reaching the maximal value at an ACh concentration of 10(-5) M (25% during 60 min incubation). The effect of ACh was not detectable in the presence of actinomycin D and puromycin. Upon removal of the nuclei from the system, preincubation with ACh produces a lowering of AChE activity in microsomes. cAMP and cGMP reduce AChE activity of microsomes in a complete system. The results obtained enable one to consider acetylcholine regulation of microsomal AChE level in nerve cells as a multi-component system. The main component in the system is a direct action of ACh on the synthesis of AChE gene product or on its processing, while the other components (cyclic nucleotides and effect of ACh itself on translation) are elements of correction. PMID- 6279209 TI - [Activation of citrate synthase of liver mitochondria by norepinephrine and cyclic AMP]. PMID- 6279210 TI - [Sensitivity of hypothalamic neurons to beta-endomorphin, met-enkephalin, and thyroliberin]. AB - A study was made of susceptibility of hypothalamic neurons to beta-endorphine, thyroliberin and met-enkephalin applied microiontophoretically. The opioids were shown to exert a primarily unidirectional effect on the same neurons irrespective of the fact that the inhibitory action of beta-endorphine was more pronounced. The nalorphine-competitive antagonist of the opiates removed the met-enkephalin induced inhibition. Unlike opioids, thyroliberin largely activated the test neurons. The possibility of neuropeptide participation in the control of gonadotropic function of the pituitary is discussed. PMID- 6279211 TI - [Effect of immunization on functional state of thyroid gland and thyroxine binding in rat organs]. AB - A number of modern research methods were used in experiments on 220 white rats to examine thyroid function at varying times after single immunization with typhoid vaccine. It was found that on the 1st and 2nd days of the observation period, only cAMP level of the thyroid parenchyma decreases. Three, four and seven days following immunization, thyroid function is drastically reduced, which manifests by a decrease in 131I uptake by the thyroid gland. Also decreased were the conversion ratio, the levels of thyroxine and triiodothyronine in blood serum and incorporation of labeled thyroxin by the heart, liver and spleen. During the productive phase of the immunogenesis (days 10, 15, 20 and 25), all the test indicators considerably increase. The phases of the reaction obtained seem likely to be a consequence of reciprocal relationship between the thyroid and sympathoadrenal systems (the inductive period), as well as by the feedback effect (the productive period of antibody formation). The reaction of thyroid function to immunization is regarded as a component of non-specific adaptation response of the body to an extreme irritant, that provides for metabolic reconstruction necessary for the synthesis of antibodies and formation of immune lymphocyte populations. PMID- 6279212 TI - High blood and urine levels of cadmium in phosphate workers: a preliminary investigation. PMID- 6279213 TI - Performance of meadow voles from sewage sludge-amended fields in swim-escape behavior trials. PMID- 6279215 TI - pH dependence of EPR spectra of nitroxides containing ionizable groups. PMID- 6279214 TI - Effects of hydroxyurea and benzo(a)pyrene on DNA synthesis in the isolated perfused rat lung. PMID- 6279216 TI - Interrelationships of glandular trichomes and cannabinoid content. II. Developing vegetative leaves of Cannabis sativa L. (Cannabaceae). AB - Gland number and cannabinoid content were quantified during ontogeny of vegetative leaves from three clones of Cannabis. Initiation of capitatesessile and bulbous glands was found to occur uniformly during leaf development. Cannabinoids were synthesized throughout leaf development as well, but at a decreasing rate. A positive correlation was found for total capitate-sessile glands per leaf as compared with total cannabinoid content of the leaf. The data also indicated that other leaf tissues in addition to the glands may contain cannabinoids. PMID- 6279217 TI - The role of presynaptic alpha-adrenoceptors in the regulation of blood pressure in the conscious rabbit. AB - 1 Changes in mean arterial pressure, heart rate and plasma noradrenaline after alpha-adrenoceptor blockade with several alpha-adrenoceptor antagonists have been studied in the conscious rabbit in order to investigate the possible role of presynaptic alpha-adrenoceptors in cardiovascular regulation. 2 Prazosin (0.05-2 mg/kg) and phentolamine (0.5-20 mg/kg) produced dose-dependent falls in mean arterial pressure and rises in plasma noradrenaline. These changes were related to the degree of postsynaptic alpha-adrenoceptor blockade determined by the pressor response to intravenous phenylephrine. 3 Similar changes in mean arterial pressure and plasma noradrenaline were observed after administration of the direct vasodilators hydralazine (1-10 mg/kg) and nitroprusside (2.5-55 microgram kg-1 min-1). 4 After baroreceptor deafferentation by sinoaortic denervation the falls in mean arterial pressure were much greater and the rise in plasma noradrenaline was markedly attenuated. 5 Yohimbine (1 mg/kg) increased mean arterial pressure and plasma noradrenaline but it was not possible to exclude the possibility that central nervous effects of yohimbine underlay the increased sympathetic activity. 6 The magnitude of the baroreflex response to changes in pressure make it unlikely that the functional significance of the presynaptic alpha-adrenoceptor can be readily determined by measurement of plasma noradrenaline in intact animals. PMID- 6279218 TI - Dual effects of catecholamines on pre- and post-junctional membranes in the dog trachea. AB - 1 Effects of noradrenaline or isoprenaline on the membrane and contractile properties of the smooth muscle cell, or on the excitatory neuro-effector transmission in the dog trachea, in vitro, were observed by use of microelectrodes and double sucrose gap methods.2 Noradrenaline (<5 x 10(-6) M) or isoprenaline (<5 x 10(-7) M) modified neither the membrane potential nor the membrane resistance. Increased concentrations of noradrenaline (>5 x 10(-5) M) depolarized and isoprenaline (>5 x 10(-7) M) hyperpolarized the membrane, and these actions were suppressed by phentolamine and propranolol respectively. Both catecholamines reduced the membrane resistance.3 Noradrenaline (5 x 10(-6) M) or isoprenaline (5 x 10(-7) M) reduced the resting tension, raised the mechanical threshold required to produce the contraction and suppressed the amplitude of phasic contractions evoked by electrical depolarization of the membrane.4 The action potential evoked by an outward current pulse in the presence of tetraethylammonium (TEA) was not affected by 5 x 10(-6) M isoprenaline, while the mechanical response was markedly suppressed.5 The excitatory junction potential (e.j.p.) evoked by electrical field stimulation was blocked by atropine. Noradrenaline (5 x 10(-7) M) or isoprenaline (5 x 10(-8) M) suppressed the amplitude of e.j.p. with no change in the membrane potential or input membrane resistance. Depression in the amplitude of e.j.ps produced by noradrenaline or isoprenaline reduced the amplitude of phasic contractions evoked by e.j.ps.6 These inhibitory actions of the catecholamines on mechanical responses and on e.j.ps were suppressed by pretreatment with propranolol (4 x 10(-6) M).7 Dog tracheal smooth muscles are innervated by cholinergic excitatory and adrenergic inhibitory systems. Electrical field stimulation produced excitation of both cholinergic and adrenergic nerve fibres, and propranolol (4 x 10(-6) M) enhanced the amplitude of e.j.p. generated by excitation of cholinergic nerves when repetitive stimulation (10 stimuli at 20 Hz) was used, but not the amplitude of the e.j.p. evoked by a single stimulus.8 5-Hydroxytryptamine (6 x 10(-6) M) produced a tonic contracture of the dog trachea. After pretreatment with atropine (4 x 10(-6) M), field stimulation (50 mus in duration and repetitive stimuli at 20 Hz) induced reversal of the contracture induced by 5-hydroxytryptamine and this was abolished by propranolol (5 x 10(-6) M).9 These results indicate that endogenous or exogenous catecholamines, in relatively low concentrations, predominantly activate beta-adrenoceptors in the pre- and post-junctional membrane in the dog trachea, and induce muscle relaxation. PMID- 6279220 TI - Scintigraphic features of superior vena cava obstruction due to substernal non toxic goitre. AB - Four men with clinical features of superior vena caval syndrome due to large substernal non-toxic multinodular goitre have undergone radionuclide superior vena caval studies. In all cases the dynamic scintigraphs showed a characteristic thyroidal configuration attributed to the definition of stretched thyroidal veins and their tributaries. Radiocontrast studies of two patients demonstrated the characteristic thyroid configuration. A radionuclide superior venacavogram taken post-thyroid lobectomy in one case showed the features had reverted to normal. PMID- 6279219 TI - The actions of pentobarbitone, procaine and tetrodotoxin on synaptic transmission in the olfactory cortex of the guinea-pig. AB - 1 It has been suggested that the depression of excitatory synaptic potentials produced by general anaesthetics can be attributed to a partial blockade of impulse conduction in the terminal branches of axons. This hypothesis has been tested by comparing the actions of pentobarbitone, procaine and tetrodotoxin (TTX) on synaptic transmission in the guinea-pig olfactory cortex. 2 Pentobarbitone (0.1-0.3mM) depressed the evoked synaptic potentials without any significant depression of impulse conduction in the afferent fibres of the lateral olfactory tract (1.o.t). It had no effect on the electrical excitability of either the l.o.t axons or the postsynaptic neurones. 3 Tetrodotoxin (TTX; 1 5x10(-8 M) slowed conduction of impulses in the l.o.t. and decreased the amplitude of the l.o.t compound action potential in proportion to the concentration applied. All concentrations of TTX elevated the electrical threshold of the l.o.t. axons and there was evidence to suggest that the threshold of the postsynaptic neurones was also elevated. The synaptic potentials were depressed in direct proportion to the depression of the l.o.t. compound action potential. 4 Procaine (0.1-0.5 mM) exhibited a pattern of activity intermediate between pentobarbitone and TTX. The most marked effect, seen at all concentrations tested, was a slowing of impulse conduction and a decrease in the electrical excitability of the l.o.t. axons. 5 It is concluded that general anaesthetics (exemplified by pentobarbitone) depress synaptic transmission by interfering with the processes involved in chemical transmission and not by blocking impulse conduction in the terminal branches of afferent nerves. PMID- 6279221 TI - Parenchymal filling of hepatic metastasis demonstrated by endoscopic retrograde cholangiography. PMID- 6279222 TI - Skin decontamination--a comparison of four methods. PMID- 6279223 TI - Does the type of suture material contribute to the strength of the lateral paramedian incision? AB - A prospective randomized trial was carried out on 231 consecutive laparotomies in which the lateral paramedian incision was used in all cases. Closure of the wound was identical except for the anterior rectus sheath where closure was randomized to (a) chromic catgut, (b) polyglycolic acid or (c) polypropylene. Follow-up for 1 year has revealed no wound dehiscences and only one incisional hernia (in the catgut group). We conclude that the lateral paramedian incision is inherently strong, and that this strength is due to splintage of the wound by the rectus abdominis muscle itself and is unrelated to the type of suture material used. PMID- 6279224 TI - Dose-related changes in vaginal cytology after topical conjugated equine oestrogens. PMID- 6279226 TI - Is bran useful in diverticular disease? PMID- 6279225 TI - Prevalence of antibodies to poliovirus in 1978 among subjects aged 0-88 years. AB - The antibody state of a population aged 6 months to 88 years to poliovirus types 1, 2, and 3 was determined by examining 919 sera collected in Lancashire, London, and southern and south-east England. In subjects aged over 2 years the immune state was surprisingly uniform, although the older patients had probably acquired practically all their antibodies as a result of natural infection and those under 16 through vaccination. at least 95% had detectable antibodies to at least one poliovirus type and about 60% to all three types, with the exception of a cohort of children born between 1963 and 1968, in whom the proportions were about 80% and 40% respectively. These children were born around the time of the changeover from inactivated to oral vaccine, when immunisation rates were low and there was confusion over the number of doses required. These results indicate that a complete course of vaccine or a booster dose at or around school-leaving age is necessary. PMID- 6279227 TI - Postexposure immunoprophylaxis against B virus infection. PMID- 6279228 TI - Failure to heal D-deficiency rickets and suppress secondary hyperparathyroidism with conventional doses of 1,25-dihydroxy vitamin D3. PMID- 6279229 TI - Burst abdomen and incisional hernia: a prospective study of 1129 major laparotomies. AB - Burst abdomen and incisional herniation are continuing problems for the general surgeon. A prospective study was carried out to define the extent of the problem. Over five years from 1975 to 1980 a total of 1129 major laparotomy wounds in adults were assessed at regular intervals for 12 months after operation. There were 19 burst abdomens (1.7%) and 84 incisional hernias (7.4%). The introduction of the mass-closure technique reduced the incidence of burst abdomen from over 3% in 1975 to 0.95% in 1979. It did not, however, improve the rate for incisional hernias, which was 7.6% in 1979. Many factors are associated with incisional herniation: old age, male sex, obesity, bowel surgery, type of suture, chest infection, abdominal distension, and, most important, wound infection. More work is needed to find the ideal method of wound closure, and efforts should be made to eliminate wound infection. PMID- 6279230 TI - Short-term prophylaxis with cefotaxime for prostatic surgery. AB - A randomised controlled trial of a new cephalosporin, cefotaxime, was carried out in men undergoing transurethral resection of the prostate. The purpose of the trial was to determine whether 48-hour prophylaxis with this new broad-spectrum, non-nephrotoxic cephalosporin would reduce postoperative bacteriuria and postoperative complications. The treated patients fared significantly better than the non-treated patients in having fewer febrile episodes, fewer episodes of tachycardia, a lower incidence of appreciable bacteriuria postoperatively, and fewer complications, and spending on average one day less in hospital. There was no difference in postoperative urea and creatinine concentrations between the groups, and no other side effects of cefotaxime occurred in this elderly population. Prophylaxis with cefotaxime would appear to make prostatic surgery safer. PMID- 6279231 TI - Lack of effect of bicarbonate on the survival of Treponema pallidum (Nichols) in vitro. PMID- 6279232 TI - Evidence in favor of a neurotransmitter role of glycine in the rat cerebral cortex. AB - In the present study we analyze whether glycine satisfies some electrophysiological and biochemical criteria to consider it as a putative transmitter in the rat cerebral cortex. Intracellular recordings from rat sensory motor cortex showed that in 15-20% of the tested neurons glycine hyperpolarized the cell membrane, decreased the firing rate and flattened the evoked EPSP-IPSP sequence by increasing the membrane conductance. The iontophoretic application of strychnine antagonized the block of 'spontaneous' firing and the membrane hyperpolarization induced by glycine. Moreover, in a group of neurons, strychnine decreased the amplitude and duration of the IPSP and brought back the membrane potential to resting values. Previously accumulated [3H]glycine and endogenous glycine were released from cortical synaptosomal preparations by depolarizing stimuli in a Ca2+-dependent way. The release pattern of glycine was qualitatively similar in cortical and in spinal synaptosomes. [14C]Glycine was rapidly synthetized from [14C]serine in cortical synaptosomal preparations, and the newly formed [14C]glycine was released by depolarizing stimuli in a Ca2+-dependent way. It is concluded that glycine, which is generally considered as an inhibitory neurotransmitter in the spinal cord, medulla and pons, may also have a transmitter role in a discrete number of cortical neurons of some mammalian species.U PMID- 6279233 TI - Barbiturate reduction of calcium-dependent action potentials: correlation with anesthetic action. AB - Calcium-dependent action potentials were recorded from mouse spinal cord neurons in primary dissociated cell culture following addition of the potassium channel blockers tetraethylammonium ion and 3-aminopyridine. The pharmacologically active barbiturates, pentobarbital and phenobarbital, but not the pharmacologically inactive barbiturate, barbituric acid, produced reversible, dose-dependent reduction of action potential duration at sedative-hypnotic and anesthetic concentrations. Pentobarbital reduced action potential duration at concentrations from 25 to 600 microM (50% reduction at 170 microM) while phenobarbital reduced action potential duration at concentrations from 100 to 5000 microM (50% reduction at 900 microM). The barbiturate concentrations which reduced calcium dependent action potential duration in this study correlate with reduction of neurotransmitter release from other neuronal preparations and with reduction of calcium uptake by synaptosomes. The results suggest that barbiturates may produce anesthesia in part by reduction of presynaptic calcium entry and consequent reduction of neurotransmitter release in addition to postsynaptic increase of membrane chloride ion conductance. Barbiturate anticonvulsant actions are probably due to postsynaptic augmentation of GABA-mediated inhibition and depression of excitatory synaptic transmission. The major difference between anticonvulsant (phenobarbital) and anesthetic (pentobarbital) barbiturates was the dose-dependency of these actions. Phenobarbital produced postsynaptic modulation of neurotransmitter responses at low concentrations and decreased calcium-dependent action potential duration and increased chloride ion conductance at high concentrations. In contrast, pentobarbital produced all actions at low concentrations. Thus for phenobarbital there would be a large therapeutic index for anticonvulsant activity compared to anesthetic activity but for pentobarbital there would be a small therapeutic index. PMID- 6279235 TI - Circadian rhythm and neural regulation of rat pineal angiotensin converting enzyme. AB - Angiotensin converting enzyme was detectable in rat pineal gland and exhibited a circadian rhythm in activity with maximum at the end of the light phase of daily photoperiod. Superior cervical ganglionectomy (SCGx) or exposure to light for 6 days increased enzyme activity and obliterated morning-evening differences, whereas injection of the beta-agonist isoproterenol depressed the high values observed in SCGx animals. These results indicate that angiotensin converting enzyme in the pineal gland is under negative control by the norepinephrine released from pineal sympathetic nerves. PMID- 6279234 TI - Selective action of piretanide on primary afferent GABA responses in the frog spinal cord. AB - The effects of piretanide, a drug which blocks active chloride transport in other systems, was examined on amino acid responses and synaptic potentials in the frog spinal cord. It was found that piretanide had no effect on GABA ventral root hyperpolarizing responses, but abolished dorsal root depolarizing responses. Dorsal root glutamate responses were little affected by piretanide. The depression of GABA responses on dorsal root ganglion cells was accompanied by a large decrease in the conductance change elicited by GABA. In addition, there was a small shift of the GABA equilibrium potential toward the resting membrane potential, which is consistent with a reduced inward pumping of chloride ions. Piretanide abolished the dorsal root potential elicited by ventral root stimulation. In addition it reduced the early part of the dorsal root potential evoked by stimulating an adjacent dorsal root but had remarkably little effect on the late phase. PMID- 6279236 TI - Negative slope conductance due to a persistent subthreshold sodium current in cat neocortical neurons in vitro. AB - The voltage dependent ionic currents of large layer V neurons of cat sensory/motor cortex were examined in an in vitro slice preparation using a single-microelectrode voltage clamp. These cells exhibit a persistent inward current in a voltage range below spike threshold. This inward current is responsible for the increase of input resistance upon depolarization seen in these cells in response to a constant current pulse and is activated at the same voltages traversed by the membrane potential between spikes during rhythmic firing. The inward current appears to be a persistent sodium current, since it is unaffected by extracellular Ba2+ or Co2+ but is blocked by extracellular TTX or intracellular QX314. PMID- 6279238 TI - Lumbar intrathecal naloxone blocks analgesia produced by microstimulation of the ventromedial medulla in the rat. AB - In lightly barbiturate-anesthetized rats. low threshold (less than 10 micro A) electrical stimulation within the rostral ventromedial medulla inhibited the tail flick response to noxious heat. Naloxone applied intrathecally at the lumbar level reversed this inhibition, but the same dose of naloxone applied to the cervical intrathecal space had no effect. Doses of naloxone 1- to 4-fold greater than the intrathecal dose did not antagonize tail-flick suppression when given systemically. Because neither systemic nor intrathecal naloxone had any effect on base-line tail-flick latencies, we conclude that the inhibition of the tail-flick response resulting from microstimulation in the ventromedial medulla is mediated by a spinal opioid synapse. PMID- 6279237 TI - Specific binding of [3H]prostaglandin E2 to rat brain membranes and synaptosomes. AB - There is saturable, reversible and specific binding for [3H]prostaglandin E2 (PGE2) to rat brain membranes. This binding is of high affinity, selectively distributed with a maximum in the hypothalamus, the amygdala and the posterior pituitary, and is associated subcellularly with the synaptosomal fraction. This specific PGE2 binding has the characteristics expected for receptors, so opening new perspectives which might clarify the role of PGs in the brain. PMID- 6279239 TI - A spinal opioid synapse mediates the interaction of spinal and brain stem sites in morphine analgesia. PMID- 6279241 TI - The influence of ACTH and corticosterone on [3H]GABA receptor binding in rat brain. AB - Bilateral adrenalectomy (ADX) induces a significant, regionally selective, increase in GABA, but not cholinergic muscarinic or alpha1-adrenergic, receptor binding in rat brain. The increase in GABA receptor binding in the midbrain occurs within 72 h of surgery, whereas that found in the corpus striatum becomes evident between 1 and 2 weeks later. These ADX-induced receptor changes are counteracted by the administration of corticosterone, a reversal which can occur within 24 h following a single administration of steroid. Unlike ADX, hypophysectomy causes a significant reduction in [3H]GABA receptor binding in these two brain areas, an action that is not reversed by corticosterone treatment. Furthermore, systemic administration of either ACTH1-39 or ACTH4-10 in unoperated animals causes an increase in midbrain and striatal GABA receptor binding similar to that observed in ADX animals. The increase in [3H]GABA binding observed after ACTH administration appears to be due to the appearance of low affinity, high capacity binding sites not observed in untreated animals. ADX had no effect on high affinity of GABA uptake, glutamic acid decarboxylase or GABA content in the brain regions where receptor modifications were noted. These findings indicate that GABA receptor binding in rat brain can be modified by changes in the circulating levels of ACTH. PMID- 6279240 TI - Some properties of solubilized GABA receptor. AB - gamma-Aminobutyric acid (GABA) receptor was solubilized from synaptic membrane of the rat brain by various detergents. Nonidet P-40, a non-ionic detergent, was found to be an effective solubilizing agent, since it caused no interference on the receptor binding assay, yielded a [3H]muscimol binding protein with a high specific activity and no aggregation, and preserved good stability of the solubilized fraction. Ammonium sulfate precipitation of the solubilized supernatant significantly increased the binding of [3H]muscimol to GABA receptor, possibly by removing heat-stable and small molecular inhibitory substances. The specific [3H]muscimol binding to the soluble fraction obtained by Nonidet P-40 treatment and subsequent ammonium sulfate precipitation, was saturable with KD 13 and 64 nM, and Bmax 3.4 and 1.8 pmol/mg protein, respectively. The enhancement of the [3H]muscimol binding by diazepam as found in synaptic membrane was also detected in the soluble fraction. Molecular weight of the [3H]muscimol binding site was determined by gel filtration on Sephadex G-200 and was calculated to be 270,000 daltons. This value was identical with that of the [3H]flunitrazepam binding site which appeared in the same solubilized fraction. These results indicate that the properties of solubilized GABA receptor are identical to those of membrane-bound GABA receptor. Furthermore, the present results suggest that both GABA and benzodiazepine receptors may reside on the same macromolecule in synaptic membrane. PMID- 6279242 TI - Evidence for increased release of prostaglandins of E-type in response to orthodromic stimulation in the guinea-pig superior cervical ganglion. AB - Prostaglandins of the E type (PGEs) stimulate cyclic adenosine 3',5' monophosphate (cAMP) biosynthesis both in isolated preparations of rat, guinea pig and rabbit superior cervical ganglia (SCG) and in calf SCG slices. Electrical stimulation of preganglionic nerve fibers of the guinea-pig SCG remarkably increased PGE release and cAMP biosynthesis. These effects were blocked by reducing the Ca2+ to Mg2+ ratio in the incubation medium. Atropine (1 microM) and phentolamine (10 microM) inhibited PGE biosynthesis and significantly reduced cAMP levels. PMID- 6279243 TI - Electrophysiological evidence for a PGE-mediated presynaptic control of acetylcholine output in the guinea-pig superior cervical ganglion. PMID- 6279245 TI - Mutual inhibition between perigeniculate neurones. PMID- 6279244 TI - Ca2+ and calmodulin-regulated endogenous tubulin kinase activity in presynaptic nerve terminal preparations. AB - Synaptosomal tubulin was shown to be the major substrate for a Ca2+-calmodulin regulated protein kinase in synaptosome soluble fractions as determined by two dimensional gel electrophoresis and peptide mapping. Ca2+ activated this endogenous tubulin kinase system in presynaptic nerve terminal preparations. The Ca2+-dependent activation of the tubulin kinase system was mediated by the Ca2+ binding protein, calmodulin. Trifluoperazine, a known inhibitor of calmodulin, significantly blocked the calmodulin-stimulated [32P]phosphate incorporation into synaptic tubulin. This inhibition of endogenous tubulin phosphorylation could be reversed by addition of exogenous calmodulin to the reaction mixture. The concentrations of Ca2+ and calmodulin required to produce a half-maximal stimulation of the tubulin kinase were 0.8 microM and 0.3 microM respectively. Greater than 70% of soluble tubulin present in the nerve terminal was phosphorylated in less than 50 s by this kinase system. Evidence is presented indicating that the synaptic Ca2+-calmodulin tubulin kinase is a distinct enzyme system from the previously described cyclic AMP microtubule-associated kinase. The anticonvulsant phenytoin inhibited the Ca2+-calmodulin stimulated phosphorylation of tubulin, and alpha- and beta-tubulin were identified as major components of previously designated synaptic phosphoprotein bands of DPH-L and DPH-M. Existence of the kinase as a calmodulin-tubulin-kinase complex is suggested from kinetic studies. The Ca2+-calmodulin tubulin kinase is very labile and specialized isolation procedures were necessary to retain activity. The activation of the tubulin kinase by Ca2+ and calmodulin may play a role in the functional utilization of tubulin in the nerve terminal and may mediate some of the effects of Ca2+ on synaptic function. PMID- 6279246 TI - A search in rat brain cortex synaptic vesicles for endogenous ligands for kainic acid receptors. AB - Conditions were found for stabilizing rat brain cortex kainic acid (KA) receptors. Such receptors had the same Hill number (about 0.6) for KA and for glutamate. The receptors were then used as detectors for endogenous ligands present in brain cortex synaptic vesicle (SV) soluble extracts. When these SV extracts were fractionated by gel filtration on Sephadex G-10, by thin-layer chromatography, or by high voltage electrophoresis, a single endogenous component, that in all cases comigrated with glutamic acid, was found. PMID- 6279247 TI - Development of spontaneous neuronal activity in the caudate nucleus, globus pallidus-entopeduncular nucleus, and substantia nigra of the cat. AB - Spontaneous single unit activity was obtained from caudate (Cd), globus pallidus entopeduncular nucleus (GP-Ento), and substantia nigra (SN) neurons in kittens of 1-60 days of age and adult cats. Five developmental trends were found in the spontaneous firing patterns of these neurons: (1) overall mean interspike intervals (ISIs) decreased with age; (2) the occurrence of neurons with shorter mean ISIs (less than 400 ms) increased with age; (3) the occurrence of neurons with burst activity increased with age; (4) burst activity became more complex with age; and (5) the rate of burst occurrence in neurons with burst activity increased with age. Neurons within each region of the basal ganglia had characteristic patterns of spontaneous activity. Furthermore, the developmental patterns of spontaneous neuronal activity were different in each structure. The spontaneous activity of GP-Ento and SN neurons matured before the spontaneous activity of Cd neurons. Thus, spontaneous firing may mature in the output nuclei of the basal ganglia prior to its maturation in the Cd. PMID- 6279248 TI - Localization of nerve growth factor bound to neurons growing nerve fibers in culture. AB - The peptide hormone, nerve growth factor, is necessary for the normal growth and development of sensory and sympathetic neurons. In culture, this hormone is also need for the regeneration of fibers from sensory and sympathetic neurons. We have needed for the regeneration of fibers from sensory and sympathetic neurons. We have determine the distribution of the nerve growth factor receptors on cells cultured from embryonic dorsal root ganglia. The binding of the radiolabeled nerve growth factor is highly selective for neurons, though some saturable binding is present on non-neuronal cells, the density of binding is less than 10% of that on neurons. By incubating cultures with different concentrations of radiolabeled nerve growth factor, we were able to quantify the type I (high affinity) and type II (low affinity) sites on neuronal somata and along growing nerve fibers. We estimate that there are 5 times more type I sites on the nerve fibers than on the cell bodies. On the other hand, type II sites appear to be uniformly distributed on nerve fibers and cell bodies. The amount of radiolabeled nerve growth factor binding was directly correlated with the extent of nerve fiber growth. However, binding to growth cones was variable and was not obviously different from the main axis cylinder of the growing nerve fiber. Finally, the saturable binding found on non-neuronal cells appears to be the same as the type II sites on neuronal cells. Bases upon our calculations, the apparent equilibrium dissociation constants for the two types of cells are the same (approximately 2 X 10(-9) M). PMID- 6279249 TI - Role of midbrain raphe nuclei in stress-, pentobarbital-, beta-endorphin-, or TRH induced changes in plasma PRL levels of adult male rats. AB - Radiofrequency lesions of either the dorsal (LD) or the median (LM) raphe nuclei of male rat mesencephalon did not modify baseline levels of plasma prolactin (PRL). However, the PRL releasing effect of 30 min of immobilization stress was suppressed in LM rats and enhanced in LD rats. The PRL releasing effect of pentobarbital (PB, 50 mg/kg, IP) or of beta-endorphin (END, 15 micrograms/rat, intracerebroventricularly, ICV) also was enhanced in LD rats. TRH (10 micrograms/rat, ICV) administered concomitantly with either PB or END, antagonized the releasing effect of the former and enhanced the releasing effect of the latter in sham operated rats. Lesions of the raphe nuclei blocked the antagonizing effect of TRH, while the enhancing effect was heightened in LD rats. These results indicate that neurons originating in the raphe nuclei are not involved in the control of baseline plasma PRL levels. They indicate, furthermore, the existence of an inhibitory pathway originating in the dorsal raphe nucleus the suppression or activation of which is, at least partly, the mechanism of PB, END or TRH effects on PRL release. The PRL releasing effect of immobilization stress seems to be under a dual, mutually antagonistic control: activating through the median and inhibitory through the dorsal nucleus. PMID- 6279250 TI - Changes in hippocampal CA1 population spikes following administration of delta-9 THC. AB - The effects of delta-9-tetrahydrocannabinol (delta-9-THC) on monosynaptic population responses in hippocampal CA1 pyramidal cells were examined in acute rats for several doses and at several post injection time periods. Delta-9-THC enhanced paired pulse inhibition in a dose-dependent manner; however biphasic dose effects were observed on cell responses during low frequency potentiation and on population spikes elicited by a conditioning stimulus. The biphasic responses were such that 2--4 mg/kg THC (IP) excited while 16 mg/kg depressed the stimulus elicited CA1 response. For low doses of THC slight biphasic time effects were observed. Evidence presented suggests that some of THC's actions on hippocampus may be due to direct or indirect actions on GABA-mediated activity. PMID- 6279252 TI - [Purification of hepatitis A virus from human feces]. AB - Hepatitis A virus was purified from on early acute phase human stool by a procedure using 10% polyethylene glycol precipitation, sepharose 2B gel filtration, isopycnic banding in cesium chloride, rate zonal separation in sucrose - 99,5% of the 280 nm absorbing fecal impurities were removed. PMID- 6279251 TI - [Aspects of megakaryocytic viral production in relation to lymphocytic leukemia in the rat]. AB - Two viral populations BL/F (EL) and BL/F (SL) were derived from RadLV-Rs by propagation in rats where they induced respectively a generalized lymphoma in 5-6 weeks or a thymic lymphoma killing the animals in 5-6 months. In both cases, 10 days after inoculation of viral extract, numerous viral particles are present in the megakaryocytes (MKC) of the bone marrow and the spleen. Our results suggested a production rather than a passive accumulation of those particles by the MKC. The kinetics of blood platelet level for both leukemias showed a thrombocytopenia corresponding with the macroscopic development of the tumor. Therefore the evolution of the blood platelet level is not related to the MKC viremia. This suggests a lack of direct effect of virus BL/F on the MKC metabolism. PMID- 6279253 TI - [Comparison of the molecular forms of exonuclease and ribonuclease in normal human spleen]. AB - Molecular forms of exonuclease and ribonuclease from normal human spleen were separated by preparative electrofocusing and characterized by their enzymatic properties. Six molecular forms (E1 to E6) of exonuclease can be separated by isoelectric focusing from human normal spleen. Properties of these molecular forms have been studied: substrate specificity, hydrolysis of paranitrophenyl thymidine-3'-monophosphate, optimum pH and heat-stability. So two enzymatic groups can be separated: - an acidic group contained forms E2, E3, E4 hydrolysing at high level the paranitrophenyl-thymidine-3'-monophosphate, with optimum pH between 5.0 and 5.5. These three forms were heat labile at 50 degrees C and belong probably to the same isoenzymatic group. - a neutral or more alkaline group was represented by the forms, E1, E5 and E6. These forms hydrolysed at very low level the paranitrophenyl-thymidine-3'-monophosphate and were different by their optimum pH and heat-stability. So, these three forms do not belong to the same isoenzymatic group. Profiles of ribonuclease showed three molecular forms R1, R2 and R3 different from exonucleases by their isoelectric point, optimum pH, substrate specificity and overall heat-stability. These ribonuclease forms differed from one another in optimum pH and probably do not belong to the same isoenzymatic group. PMID- 6279254 TI - Effect of Fe2+ and ascorbic acid on acid phosphatases from rat bone. PMID- 6279255 TI - Alkaline phosphatase inhibition by parathyroid hormone and isoproterenol in a clonal rat osteosarcoma cell line. Possible mediation by cyclic AMP. AB - The effect of parathyroid hormone (PTH 1-34 bovine) on alkaline phosphatase activity was investigated in an osteoblast-like clonal cell line derived from rat osteosarcoma (ROS 17/2). ROS 17/2 alkaline phosphatase resembled the bone enzyme in levamisole sensitivity and electrophoretic mobility but differed in heat stability. The specific activity of ROS 17/2 alkaline phosphatase increased with time in culture. This increase was inhibited by PTH (1-34) and (-)-isoproterenol in a dose-dependent manner starting at near-physiological hormone concentrations. The ID50 values were 0.02 nM for PTH (1-34) and 1.7 nM for isoproterenol. The two hormones stimulated ROS 17/2 adenylate cyclase, albeit at higher concentrations: Km values were 13 nM for PTH (1-34) and 16 nM for isoproterenol. The rise in alkaline phosphatase was also inhibited by dibutyryl cyclic AMP and 8-bromocyclic AMP (0.1 mM). These findings further document the osteoblastic properties of the ROS 17/2 osteosarcoma cell line, suggest that PTH inhibition of alkaline phosphatase represents a physiological response to the hormone in these cells, and implicate cyclic AMP as a mediator of this PTH effect. PMID- 6279256 TI - Influence of pregnancy on immunoreactive parathyroid hormone levels. PMID- 6279257 TI - Effects of cyclic nucleotides on hemoglobin synthesis in fetal calf liver cells in culture. PMID- 6279258 TI - Isolation and characterization of subcellular membranes from canine stomach smooth muscle. AB - Subcellular membrane fractions were isolated from the circular muscle of the corpus of canine stomach by differential and isopycnic sucrose density gradient centrifugation. Differential centrifugation gave a mitochondrial fraction enriched (fourfold) in cytochrome c oxidase and a microsomal fraction enriched (fourfold) in 5'-nucleotidase and NADPH-cytochrome c reductase over postnuclear supernatant. On the basis of a study using continuous gradient, a discontinuous sucrose density gradient was prepared to yield F1 to F5 fractions. The F3 fraction at the interface of 18-32% (w/w) sucrose was maximally enriched (13 fold) in 5'-nucleotidase. The fraction contained very low levels of cytochrome c oxidase but did contain NADPH-cytochrome c reductase (eightfold enrichment). The F4 fraction, at the interface of 32-40% (w/w) sucrose, was maximally enriched in NADPH-cytochrome c reductase (12-fold) and cytochrome c oxidase (6-fold). The distribution of the azide-insensitive. ATP-dependent Ca2+ uptake correlated very well with that of 5'-nucleotidase but less well with NADPH-cytochrome c reductase and not at all with cytochrome c oxidase. Sodium azide and ruthenium red inhibited the ATP-dependent Ca2+ uptake by the mitochondrial fraction and postnuclear supernatant, but not by the F3 fraction. ATP-dependent Ca2+ uptake by the F3 fraction was inhibited by calcium ionophores A23187 and ionomycin, but not by the sodium ionophore, monensin. These results are consistent with the hypothesis that the plasma membrane plays a major role ih regulating intracellular Ca2+ concentration in canine corpus circular muscle. PMID- 6279259 TI - Ba2+ ions block K+-induced contractures by antagonizing K+-induced membrane depolarization in frog skeletal muscle fibres. AB - The effects of Ba2+ ions on twitches, K+-induced contractures, and on intracellularly recorded membrane potentials (Em) and depolarizations of frog skeletal muscle fibres were investigated. Exposure of toe muscles to choline- Ringer's solution with 10(-3) M Ba2+ with Ca2+ (1.08 mM) eliminated or very greatly reduced contractures produced by 60 mM K+. In contrast, not only did the same concentration of Ba2+ ions fail to depress the twitch tension of isolated semitendinosus fibres when added to Ringer's with Ca2+, but it even restored twitches that had been eliminated in a zero Ca2+ Ringer's solution. The resting Em of sartorius muscle fibres in choline--Ringer's solution was reduced about 20 mV by 10(-3) M Ba2+. This Ba2+ ion concentration also antagonized the K+-induced depolarization. Thus in the presence of 1 mM Ba2+, 20 mM K+ hyperpolarized rather than depolarized the fibres and 60 or 123 mM K+ produced only very slowly developing, small depolarizations. These results suggest that the loss of the K+ induced contracture in choline-Ringer's caused by Ba2+ ions is due to an inhibition of the K+-induced depolarization. The latter result is consistent with previous findings of other workers that Ba2+ ions block membrane K+ channels. PMID- 6279260 TI - Response of adrenal 3beta-hydroxy-delta5-steroid dehydrogenase to adrenocorticotropin treatment in thiouracil-fed male mice. AB - A number of parallels can be drawn between the reported endocrine status of thiouracil-fed young rodents and that of aged animals, particularly with regard to the hypothalamus-pituitary-adrenal axis. Since the activity of the adrenal steroidogenic enzyme 3beta-hydroxy-delta5-steroid dehydrogenase (3beta-HSD) has been shown to be depressed in aged rats and mice, the present study was done to determine whether exposure of young mice to thiouracil had a similar effect on adrenal 3beta-HSD activity. Feeding the goitrogen thiouracil at 0.25% (w/w) of the maternal diet from conception, and keeping it 0.25% of the offsprings' diet after weaning, significantly elevated activity of 3beta-HSD per gram of adrenal gland above control levels in 4-month-old mice, perhaps to compensate for depressed adrenal mass. Daily subcutaneous injections of physiological saline (0.9%) for 4 days was sufficient to increase 3beta-HSD activity per gram of adrenal tissue in euthyroid (P less than 0.05) but not in thiouracil-fed mice. Subcutaneous administration of ACTH (2 IU daily for 4 days) significantly increased adrenal 3beta-HSD activity to comparable levels in thiouracil-fed and euthyroid animals. Thus, thiouracil enhances the activity of 3beta-HSD per gram of adrenal tissue and does not prevent response of enzyme activity to exogenous ACTH. PMID- 6279261 TI - Factors affecting recovery of latent herpes simplex virus from human trigeminal ganglia. AB - The rate of recovery of herpes simplex virus (HSV) from human trigeminal ganglia explant monolayers is affected by two factors: (1) time elapsed from the death of an individual to the establishment of in vitro culture of ganglia and (2) surface area onto which ganglia are explanted. Spontaneous reactivation of HSV from human trigeminal ganglia can be maximized when ganglia are obtained within 12 h of death and explanted into surface area of 250 cm2. Viruses isolated by explantation of human trigeminal ganglia were found to be uniformly HSV type 1 by restriction enzyme analysis. PMID- 6279262 TI - Restriction fragment analysis for differentiation of indistinguishable temperate coliphages. AB - DNA of lambda and seven related phages was digested with restriction endonuclease, EcoRI. Seven different fragment patterns were observed, only two of the eight phages showing identical profiles. Restriction enzyme fragment analysis is thus shown to be a sensitive tool for the differentiation of biologically indistinguishable phages. PMID- 6279263 TI - Antiviral activity of antibiotic-producing marine bacteria. AB - The stability of poliovirus 1 in estuarine water and sediment was examined. The present data indicated that a 2 log reduction in virus titer at 15 degrees c occurred within 6-7 days in water samples taken from estuarine waters on both sides of the Atlantic Ocean. The antiviral effect decreased significantly when the seawater was subjected to autoclaving but not when it was filtered. That the antiviral activity activity of the seawater was related to the growth activities of microorganisms was corroborated by the isolation of antibiotic-producing marine bacteria that had marked activity against poliovirus (net inactivation greater than or equal to 2 logs within 6-8 days). These organisms retained this activity following repeated subcultivation on laboratory media. Since comparable inactivation rates were observed in cell-free filtrates from these marine strains, extracellular products appear to be involved in the virus-inactivation process. Other enteric viruses, Coxsackie B-5 and ECHO-6, were also inactivated by these marine bacteria. The addition of sediment to natural seawater increased the length of poliovirus survival more than three times over that in seawater alone. However, this was not found under sterile conditions, suggesting that the sediment can protect the viruses from inactivation by the marine microflora. PMID- 6279264 TI - Electrocardiography, cardioenzymes and myocardial imaging to detect perioperative myocardial infarction. AB - Serial electrocardiography, technetium-99m pyrophosphate scintigraphy and measurement of myocardial creatine kinase (CK2) and lactic dehydrogenase isoenzyme activity (specifically the LD1 to LD2 ratio) were evaluated prospectively in 26 patients who underwent aortocoronary bypass grafting and 11 patients who underwent valvular or other thoracic surgery, as methods of diagnosing perioperative myocardial infarction. Of the 26 patients who had aorto coronary bypass grafting, 7 (group 1) had myocardial infarction perioperatively; of these, only 2 had positive results from all four diagnostic tests. The other 19 patients (group 2) had no perioperative myocardial infarction and the only diagnostic method yielding positive results was the LD1 to LD2 ratio, in 7 of the 19. In the 11 patients who did not undergo aortocoronary bypass grafting (group 3), only 1 patient had myocardial infarction perioperatively; results from the measurement of CK2 isoenzyme activity and LD1 to LD2 ratio were positive while electrocardiography and 99mTc pyrophosphate scintigraphy yielded negative results. The respective sensitivity (%) and specificity (%) of the four diagnostic methods were as follows: electrocardiography: 38, 100; 99mTc pyrophosphate scintigraphy: 88, 100; LD1 to LD2 ratio: 100, 68; CK2: 38, 100. When the CK2 criterion was redefined, using patients in group 2 as controls, the sensitivity and specificity of this method became 100 and 90 respectively. The authors conclude that 99MTc pyrophosphate scintigraphy and measurement of myocardial creatine kinase isoenzyme are of comparable value and are the most reliable indicators of perioperative myocardial infarction. PMID- 6279265 TI - Cytomegalovirus in the semen. PMID- 6279267 TI - Reflections on Alzheimer's disease. AB - As longevity increases, society will face a silent epidemic of idiopathic dementias. The concept, Alzheimer's disease, reflects a cumbersome and vaguely defined cluster of signs, symptoms and other variables which might more appropriately be labelled as the idiopathic dementias, Alzheimer-type or IDAT. Diagnosis, which is made by exclusion and treatment, primarily custodial, demonstrates the complex nature and unfortunate prognosis of the problem. Dramatic progress, nevertheless, has been made in various scientific aspects of the issue, namely, in histology, genetics and neurochemistry. The resulting evidence warrants further speculation on the role of central cholinergic neurotransmission in cognitive functioning. PMID- 6279266 TI - Neonatal herpes simplex. PMID- 6279268 TI - Viral leukoencephalomyelitis-arthritis of goats. PMID- 6279269 TI - Phase III study of CCNU, cyclophosphamide, adriamycin, vincristine, and VP-16 in small-cell carcinoma of the lung. AB - A comparison of CTX + CCNU and CTX + CCNU + procarbazine as initial systemic treatment was made in 440 evaluable patients with small-cell carcinoma of the lung. The overall response rate for patients receiving the three-drug combination was 57% (11% CR) compared with 44% (9% CR) for the two-drug combination. Median survival times were similar, 27 (with procarbazine) and 29 weeks (without procarbazine). Nonresponders to the initial treatment received ADR, ADR + VCR, and ADR + VP-16 with overall response rates of 14% (2 of 14), 27% (9 of 33), and 30% (15 of 51), respectively. Median survivals for nonresponders, as measured from day 42, were 15 weeks (ADR), 21 weeks (ADR + VCR), and 22 weeks (ADR + VP 16). Responders to the initial treatment either continued on the initial therapy or received a non-cross-resistant combination chemotherapeutic regimen (ADR + VCR) alternating with the initial therapy. There is also the suggestion that responders who received the cycled therapy after day 42 survived significantly longer than responders who did not switch treatments until relapse, 38 vs. 29 weeks. PMID- 6279270 TI - Oat-cell carcinoma of the thymus. AB - Oat-cell carcinoma of the thymus gland is exceedingly rare as a primary lesion, and only a few cases have been reported. We report a case in which a neoplasm interpreted as an oat-cell carcinoma was intimately associated with and arose in transition from a carcinoid tumor of the thymus. Ultrastructural features of this lesion were unusual compared with previous descriptions of thymus neuroendocrine neoplasms. PMID- 6279271 TI - Primary anterior mediastinal seminoma. AB - A review of the Mayo Clinic experience with primary anterior mediastinal seminomas involved 17 patients who had pure anterior mediastinal seminomas and four who had mixed germ-cell tumors containing seminomas. At follow-up, of the 17 patients with pure anterior mediastinal seminoma, nine had no evidence of disease and eight had died of metastatic disease. Of the four patients with mixed germ cell tumor containing seminoma, two were alive at follow-up and two had died of metastatic disease. In the group with pure anterior mediastinal seminoma, these factors seemed to have been associated with a greater potential for progression of disease: older than 35 years of age, presentation with fever, superior vena caval syndrome, supraclavicular or cervical adenopathy, and roentgenographic evidence of hilar disease. PMID- 6279272 TI - Intestinal mucinous substances in gastric intestinal metaplasia and carcinoma studied by immunofluorescence. AB - Immunofluorescent studies using specific antisera against intestinal mucins revealed small intestinal mucin antigen (SIMA) and large intestinal mucin antigen (LIMA) in areas of intestinal metaplasia of the stomach. In 25 gastric carcinomas studied, both these antigens were detected in seven carcinomas, SIMA only was present in four and LIMA in only four cases; the antigens could not be detected in ten of the carcinomas. In 21 of the 25 gastric operation specimens, including the 15 carcinomas positive for intestinal mucin antigens, there was evidence of chronic gastritis and intestinal metaplasia. These immunohistologic observations confirm the results of recent histochemical studies that both small and large intestinal type mucins are present in intestinal metaplasia and gastric carcinomas. Our findings provide further evidence that at least a proportion of gastric carcinomas may supervene on intestinal metaplasia. The absence of one or both antigens in gastric carcinomas may indicate stages of dedifferentiation or alternatively differences in histogenesis. PMID- 6279274 TI - Malignant mixed tumor of the skin: malignant chondroid syringoma. AB - Malignant tumors of sweat gland origin are rare. Probably one of the rarest types, still poorly understood, is the so-called malignant chondroid syringoma or malignant mixed tumor of the skin. A case of malignant chondroid syringoma is presented. Ultrastructural study proves very useful for the differential diagnosis with other chondroid or chordoid tumors. PMID- 6279273 TI - Nodular (pseudosarcomatous) fasciitis, a nonrecurrent lesion: clinicopathologic study of 134 cases. AB - Clinicopathologic review of 134 patients originally diagnosed as having nodular (pseudosarcomatous) fasciitis is presented. In 114 patients with 116 lesions, no recurrence of the lesion was noted. Of the 114 patients, 85% were younger than 50 years of age, and the forearm and arm were the most common sites of presentation. Nonrecurrent lesions rarely exceed 4 cm and 71% were smaller than 2 cm. In at least six instances, incompletely resected lesions never recurred. Though all lesions were histologically reminiscent of reparative mesenchymal tissue, four subtypes--the reactive type, the densely cellular type, those with osteoid or cartilaginous metaplasia, and the so-called proliferative fasciitis--were distinguished from the majority of lesions that conform to the description given by Kornwaler. Recurrence of the tumor was noted in 18 patients. Fifteen of 18 lesions recurred within two years, and two more recurred at 30 months following initial excision. In all these cases, review of the histology and clinical course led to a revision of the original diagnosis. The greatest number of errors was made in incorrectly classifying of inflammatory fibrous histiocytoma. Recurrence of a lesion originally diagnosed as nodular fasciitis should lead to a careful reappraisal of the pathologic findings. PMID- 6279275 TI - The natural history of colorectal carcinoma in adolescents. AB - The symptoms, histology, extent, and course of disease of 24 adolescents with colorectal carcinoma who were admitted to St. Jude Children's Hospital between 1964 and 1980 are presented. Twenty of the patients were referred between October 1974 and June 1980. Most patients presented with vague abdominal complaints. Twenty-one of the 24 patients had poorly differentiated mucin-producing adenocarcinoma. Extensive disease at diagnosis and unresponsiveness to medical management was reflected in the eight-month median survival from diagnosis. Only two of the 24 patients survive free of disease 15 and 130 months from diagnosis. Two other patients survive with disease at four and 24 months. PMID- 6279276 TI - A multivariate analysis of the prognosis after surgical treatment of malignant soft-tissue tumors. AB - Ninety-seven patients who had received their primary and definitive surgical treatment for a soft-tissue sarcoma during the years 1956--1976 were studied with respect to local recurrence, metastasis, and survival. The aim of the treatment was to eradicate the tumor while preserving good function. The influence of host and tumor properties and different diagnostic and surgical procedures on the prognosis was studied by means of nonparametric multivariate analysis. Ninety four percent of the tumors were located in the extremities. Malignant fibrous histiocytoma was the most common histologic type. A four-grade scale of histologic malignancy was used. Eighty-eight percent of the tumors were Grade III or IV. In 85% of the patients with an extremity lesion, local extirpation was carried out. The overall local recurrence rate was 21.7% and in 76 patients who underwent an adequate surgical procedure it was 6.6%. The overall five-year survival rate was 59%. The survival depended on the local control of the primary tumor, which was related to the adequacy of the surgical procedure and to the histologic grade of malignancy. No patient with a Grade I or II tumor died. The five-year survival rate for patients with Grade III tumors was 68% and for patients with Grade IV tumors 47%. PMID- 6279277 TI - Chromosome number 11 and Wilms' tumor. AB - Cytogenetic studies have been carried out on cells derived from two Wilms' tumors in vitro. Both tumors had a diploid chromosome range. One tumor was shown to have a definite stemline; 46,XY,4p+,del(9)(q22),11p-q,11p+, and the other range of variation chiefly involving chromosomes No. 11, 4, 7, and 2; most changes in chromosome No. 11 took the form of deletions of the short arm. High resolution chromosome analysis of peripheral blood lymphocytes of the two patients revealed apparently normal karyotypes. These findings suggest that changes in the short arm of chromosome No. 11 are important in the development of Wilms' tumor in normal individuals. This association is reinforced by the fact that patients with spontaneous aniridia with a 1 in 3 risk of Wilms' tumor have been reported as having a specific 11p13 deletion. PMID- 6279278 TI - Cancer nursing and the law: valuing the missed opportunity for treatment. The James case. PMID- 6279279 TI - Selective activation of some dihydrodiols of several polycyclic aromatic hydrocarbons to mutagenic products by prostaglandin synthetase. AB - The ability of prostaglandin synthetase (PGS) to cooxidize benzo(a)pyrene, benzo(a)anthracene, chrysene, and several of their dihydrodiol derivatives to mutagenic products was tested with Salmonella typhimurium strains TA98 and TA100. The microsomal fraction of ram seminal vesicles, a known source of PGS, in the presence of the PGS substrate arachidonic acid, metabolized benzo(a)pyrene-7,8 dihydrodiol, benzo(a)anthracene-3,4-dihydrodiol, and chrysene-1,2-dihydrodiol to mutagenic products. This activity was inhibited by the PGS inhibitor indomethacin. Unlike the PGS system, however, a cytochrome P-450-reduced nicotinamide adenine dinucleotide phosphate-dependent system, present in an Aroclor 1254-induced rat liver 9000 x g supernatant fraction, also activated the parent compounds [benzo(a)pyrene, benzo(a)anthracene, chrysene] and several other benzo(a)anthracene dihydrodiols (the 1,2-dihydrodiol, the 8,9-dihydrodiol, and the 10,11-dihydrodiol). The chrysene trans-3,4, trans-5,6, and cis-5,6 diols were not activated to mutagens by either system. Thus, the PGS system appears to be more selective than does the cytochrome P-450 system in the activation of polycyclic aromatic hydrocarbons to mutagenic products, activating only those dihydrodiols with adjacent double bonds in the bay region from which the bay region diol-epoxides are formed. PMID- 6279281 TI - Immune selection of tumor cell variants in chickens bearing tumors induced by avian sarcoma virus. AB - We have compared avian sarcoma cells, cultured from tumors at various stages of growth, in terms of their ability to synthesize infectious progeny virus and to express antigens that are reactive with the sensitized lymphocytes of tumor bearing hosts. Cell-mediated immunity in chickens bearing tumors induced by avian sarcoma viruses was monitored by each of a target cell cytotoxicity test and an antigen-driven lymphocyte stimulation assay. Our results show that it is only those tumor cells which have been derived from progressively growing neoplasms that are able to synthesize infectious progeny virus and to specifically interact with the sensitized lymphocytes of tumor-bearing hosts. In contrast, cells from regressing tumors do not express relevant antigens to the same extent as do progressors, and they synthesize noninfectious particles only. Experiments on cellular outgrowths, derived from the plating at limited dilution of progressively growing tumor cells, revealed that such producers of defective virus are present at the earliest stages of tumor growth. Both these cells and regressing tumor cells are poorly stained (about 10%) in indirect immunofluorescence tests by antiserum against viral envelope glycoprotein, whereas tumor cells from progressing neoplasms react well (about 60 to 85%). These results suggest that tumor cells which are found in regressing neoplasms are selected out by a functional immune response directed against cells which are efficient producers of progeny virus. PMID- 6279280 TI - Implementation of micromethods to resolve problems of human breast tumor heterogeneity in analysis of cyclic 3':5'-nucleotide phosphodiesterase. AB - Individual human infiltrating ductal carcinomas and fibroadenomas were sectioned frozen to yield an alternating sequence of stained and lyophilized material. Stained preparations were used as references permitting microdissection of regions of tumor involvement in the corresponding dried sections. Tissues quantities of 5 to 25 micrograms dry weight were incubated under mineral oil in reaction volumes of 5 microliters and analyzed for cyclic adenosine 3':5' monophosphate phosphodiesterase (PDE). The observed affinity constants for the 27,000 x g soluble PDE from benign tumors were 4.7 and 49.9 microM, while those for malignant tumors were 6.3 and 28.5 microM. The soluble enzyme of both tumor types eluted in three peaks on DEAE-Sephacel microcolumns. Both tumor types possessed a PDE activator eluting at 350 mM NaCl, although endogenous PDE activities were unaffected by additions of either this activator or 200 microM ethylene glycol bis(beta-aminoethyl ether)N,N,N',N'-tetraacetic acid. Individual microsections of benign tumors contained total PDE levels 2-fold higher than those of malignant tumors. Homogenates prepared from pooled microsections of the same tumors possessed only one-half of the total activity. Differential losses of enzyme in various preparation schemes as well as the use of tumor samples differing in cell density were suggested to account for some of the apparently conflicting literature values for breast tumor PDE. PMID- 6279282 TI - Benzo(a)pyrene phenol production by perfused rat liver and its inhibition by ethanol. AB - A rapid and inexpensive method has been developed to estimate rates of benzo(a)pyrene phenol production by perfused rat liver. This method is based on the measurement of benzo(a)pyrene phenols utilizing a simple fluorometric procedure. Within 2 to 3 min after infusion of benzo(a)pyrene bound to serum albumin, phenols are excreted into the perfusate, primarily as glucuronide and sulfate conjugates. Maximal rates of phenol release were 8 to 10 nmol/g/hr in livers from control rats and 40 to 42 nmol/g/hr in livers from 3 methylcholanthrene-treated rats. Fasting of 3-methylcholanthrene-treated rats for 24 hr prior to perfusion experiments did not affect either the rate of phenol production or the extent of their conjugation. Ethanol (20 mM) inhibited rates of phenol formation by 50% in livers from fasted, 3-methylcholanthrene-treated rats but had no effect on benzo(a)pyrene hydroxylase activity in isolated hepatic microsomes. These data indicate that ethanol inhibits phenol formation from benzo(a)pyrene in intact liver, probably by diminishing the supply of the cofactor reduced nicotinamide adenine dinucleotide phosphate. PMID- 6279284 TI - Conversion of malignant murine embryonal carcinomas to benign teratomas by chemical induction of differentiation in vivo. AB - PCC4azal embryonal carcinoma tumors were grown in strain 129 mice by s.c. transplantation. When palpable, the tumors were treated with a combination of retinoic acid and dimethylacetamide. In vitro, this embryonal carcinoma cell line shows minimal spontaneous differentiation and is exquisitely sensitive to retinoic acid and/or dimethylacetamide induction of differentiation. Ten daily 20 microliter intratumor injections of a solution of 10 mg retinoic acid per ml of dimethylacetamide resulted in nearly complete induction of morphological differentiation mainly into neuroepithelial and glandular derivatives. Control tumors showed minor spontaneous differentiation. Differentiation was associated with decreased tumor growth rate, decreased mitotic index, decreased extent of necrosis, and increased survival time of the hosts. In 4 of 18 cases, long-term survival of the hosts was effected by a complete differentiation of the malignant embryonal carcinoma tumors into benign teratomas. Retinoic acid:dimethylacetamide was also effective in inducing differentiation with the same dosage and schedule when administered systemically, i.e., i.p. or s.c. PMID- 6279283 TI - Effect of magnesium content on density-dependent regulation of the onset of DNA synthesis in transformed 3T3 cells. AB - A spontaneously transformed clone of BALB/c 3T3 cells became more transformed after more than 90 passages as indicated by increased rounding of cells, multiplication to a higher saturation density, and increased ability to form colonies when suspended in agar. When the extracellular concentration of Mg2+ was sharply reduced, the highly transformed cells flattened, assumed the shape of nontransformed cells, and became regularly arranged in cohesive arrays. If crowded when deprived of Mg2+, they lost more intracellular Mg2+ than did nontransformed and early passage-transformed cells and remained at constant cell density for at least 10 days. The intracellular content of neither Na+ nor K+ changed consistently with Mg2+ deprivation, but the Ca2+ content increased more than 2-fold. The sensitivity of the onset of DNA synthesis to inhibition by Mg2+ deprivation increased with the extent of crowding of the cultures. This was demonstrated by varying population density within a single culture dish as well as from culture to culture. The loss of intracellular Mg2+ in low concentrations of extracellular Mg2+ increased with cell crowding as did the inhibition of DNA synthesis per fractional loss of intracellular Mg2+. Neither deprivation of K+ or Ca2+ nor addition of cyclic adenosine 3':5'-monophosphate produced a density dependent inhibiton of DNA synthesis. The results indicate that a reduction of the Mg2+ content of highly transformed cells restores density-dependent inhibition of the onset of DNA synthesis, which is a characteristic property of nontransformed cells. The differences in Mg2+ retentiveness with population density may reflect differences of intracellular distribution and binding of Mg+, which could in turn explain some of the regulatory effects of population density on metabolism and growth. PMID- 6279285 TI - Metabolism of benzo(a)pyrene in epidermal keratinocytes and dermal fibroblasts of humans and mice with reference to variation among species, individuals, and cell types. AB - The metabolism of benzo(a)pyrene (BP) in epidermal keratinocytes and dermal fibroblasts of humans and mice was investigated with emphasis on variation among species, individuals, and cell types. Human epidermal and dermal cells were isolated from the skin of normal subjects by trypsinization at 4 degrees overnight, followed by separation of the epidermis from the dermis with forceps. In confirmation of previous studies metabolic activity of human epidermal cells on BP was consistently demonstrated by cell-mediated assay, in which V79 Chinese hamster cells were plated on top of sheets of epidermal cells and treated with BP for 48 hr. Mutation of the V79 cells, measured as ouabain resistance, was induced in a dose-related fashion, although the extent of induced mutation varied from 5 to 22 ouabain-resistant colonies per 10(6) survivors/10 microM BP in cultures derived from different individuals. The most striking observation was that human dermal fibroblasts did not activate BP to a form that was mutagenic to cocultured V79 cells. This was observed without exception in all nine cultures of dermal fibroblasts and the one culture of embryo fibroblasts (MR-90) tested. Analysis by high-pressure liquid chromatography indicated that human epidermal and dermal cells both metabolized BP, producing almost the whole series of known metabolites of BP. The amount of BP 7, 8-dihydrodiol, a proximate metabolite of BP, produced by human dermal cells varied from 0.2 to 2.7% of the total BP added and seemed to be enough to induce mutation. Furthermore, human dermal cells not necessarily activated exogenously added BP 7,8-dihydrodiol to a form being mutagenic to V79 cells. These observations suggest that further metabolism of BP 7,8-dihydrodiol is partially or entirely blocked in human fibroblasts. In contrast to human fibroblasts, mouse fibroblasts isolated from the dermis of embryos did activate BP and induced mutation in cocultured V79 cells to a higher extent than did mouse epidermal cells, indicating interspecies variation in metabolic activation of BP between human and mouse fibroblasts. PMID- 6279287 TI - Dome formation by a retrovirus-induced lung adenocarcinoma cell line. AB - A cell line has been established from an alveologenic lung adenocarcinoma that develops in newborn NFS/N mice after i.p. inoculation with a carcinoma-inducing retrovirus. A clonal derivative of this cell line, 3041, forms adenocarcinomas when injected into syngeneic weanling mice and in culture retains several properties typical of alveologenic type 1 cells. These include morphological details, such as tight junctions and interdigitating microvilli in the areas of cell-cell contact, as well as the ability to transport fluid in culture. Fluid transport becomes evident in confluent cultures by the formation of domes or hemicysts. The formation of domes, which is considered to reflect a differentiated function of these secretory epithelial cells, can be enhanced more than 50-fold by treatment with dibutyryl cyclic adenosine 3':5'-monophosphate but only 5- and 3-fold by sodium n-butyrate and dimethyl sulfoxide, respectively. It is anticipated that the secretory alveologenic cell line 3041 described here will be useful for studies of both its pathological and its physiological properties. PMID- 6279286 TI - Induction of cytotoxicity, mutation, cytogenetic changes, and neoplastic transformation by benzo(a)pyrene and derivatives in C3H/10T1/2 clone 8 mouse fibroblasts. AB - Benzo(a)pyrene (BaP), a series of its metabolic derivatives, and benzo(e)pyrene, a very weakly carcinogenic isomer, were tested for their biological effects on transformable C3H/10T1/2 cells. These cells were used as targets in a series of assays designed to measure oncogenic transformation, mutation to ouabain resistance, cytotoxicity, and induction of cytogenetic changes, as evidenced by chromosomal aberrations and sister chromatid exchange. Of all the compounds tested, only the parent hydrocarbon, BaP, an (+/-)-trans-7,8-dihydroxy-7,8 dihydrobenzo(a)pyrene were found to be significantly active in producing transformation and cytogenetic alterations. BaP, (+/-)-trans-7,8-dihydroxy-7,8 dihydrobenzo(a)pyrene, and (+/-)-7 alpha, 8 beta-dihydroxy-9 beta, 10 beta-epoxy 7,8,9,10-tetrahydrobenzo(a)pyrene, however, were all effective inducers of mutation in C3H/10T1/2 cells. (+/-)-7 alpha, 8 beta-Dihydroxy-9 beta, 10 beta epoxy-7,8,9,10-tetrahydrobenzo(a)pyrene was the most potent agent in tests for cytotoxicity, Benzo(e)pyrene was inactive in all assays examined. Among the compounds tested, there was a correlation between the ability to induce cytogenetic changes and the ability to produce mutation and transformation. These results support the demonstrated role of (+/-)-trans-7,8-dihydroxy-7,8, dihydrobenzo(a)pyrene as a proximal carcinogenic form of BaP and illustrate the utility of the C3H/10T1/2 cell system as an important tool for the detection of genotoxic damage by carcinogenic chemicals. PMID- 6279288 TI - Protective effects of S-2-(3-aminopropylamino)ethylphosphorothioic acid against radiation damage of normal tissues and a fibrosarcoma in mice. AB - S-2-(3-Aminopropylamino)ethylphosphorothioic acid (WR-2721) was investigated for its protective effect against radiation-produced damage of jejunum, testis, lung, hair follicles, and a fibrosarcoma of C3Hf/Kam mice. Most of these tissues were radioprotected, and the degree of radioprotection depended on the dose of WR-2721 and the time interval between administration of WR-2721 and radiation treatment. WR-2721 increased resistance of jejunal epithelial cells and spermatogenic cells to single doses of gamma-rays by factors of 1.64 and 1.54, respectively. Protection against hair loss was less pronounced; the dose-modifying factor here was 1.24. The radiation-induced acute damage of the lung expressed by the increased formation of tumor nodules in the lung was not decreased by treatment of animals with WR-2721 before radiation. In contrast, WR-2721 augmented the radiation-induced enhancement of metastasis formation in the lung. WR-2721 protected fibrosarcoma micrometastases in the lung against therapeutic effect of radiation by a factor of 1.238. In contrast, this compound had no effect on the therapy of an 8-mm fibrosarcoma growing in the legs of mice. PMID- 6279289 TI - Induction of sister chromatid exchange by polyoma large viral tumor antigen in transformed rat fibroblasts. AB - The frequency of sister chromatid exchange (SCE) was determined in rat fibroblasts transformed by wild-type polyoma virus or by a mutant temperature sensitive for viral large tumor antigen function (ts-a). Elevated SCE frequencies were observed in two wild-type transformed cell lines growing at 37 degrees and in four ts-a-transformed lines upon growth at the permissive temperature for large viral tumor antigen (33 degrees). The increase in SCE frequency in ts-a transformed cells at 33 degrees was reversed by growth at 39 degrees (nonpermissive for T-antigen function). An increase in SCE at 33 degrees was not observed in untransformed cells or in a ts-a-transformed cell line which makes a defective large viral tumor antigen. These results suggest that large viral tumor antigen can induce SCEs. Since large viral tumor antigen is also responsible for amplification of integrated viral DNA sequences (4), we tried to correlate this phenomenon with the increased SCE frequency. However, increasing SCE artificially by growing cells in the presence of 12-O-tetradecanoylphorbol-13-acetate did not result in amplification of integrated viral DNA in the absence of large viral tumor antigen function. Thus, there is no simple causal relationship between increased SCE and amplification. PMID- 6279290 TI - Establishment and characterization of SV40-transformed human breast epithelial cell lines. AB - Human breast epithelial cells cultured from milk have been transformed with SV40. Indirect immunofluorescence tests using monoclonal antibodies show that cells from clones grown in soft agar have SV40 large T-antigen in their nuclei and epithelia-specific tonofilament antigens on their intermediate filaments. In primary cultures of milk epithelial cells, the tonofilaments form a characteristic delicate basketwork throughout the cytoplasm, but in the SV40 transformed epithelial cell strains, the filament network is grossly distorted. Insulin, hydrocortisone, and serum stimulate the growth of the cell strains. At passages 8 to 11, the cell strains become quiescent and usually die. One cell strain survived this crisis period and gave rise to the fR series of cell lines. Most cell lines have a cuboidal morphology and react with a monoclonal antibody that recognizes a differentiation antigen on the membranes of breast epithelia. Line fR2 expressed the highest level of this antigen whereas fR5, the only fR line isolated with fusiform morphology, had relatively little. The in vitro transformed lines may be related to the two dominant epithelial cell types seen in primary milk cultures and could be useful for studying the relationship between transformation and differentiation in human mammary epithelial cells. PMID- 6279291 TI - Diphtheria toxin treatment of human advanced cancer. AB - Purified diphtheria toxin was administered i.v. to 50 patients with advanced solid tumors. The doses of toxin varied according to the different immunological status of the patients against diphtheria. The prominent toxic effects of the treatment were transient peripheral neuropathy observed in two nonimmune patients and fever (37-41 degrees) which occurred in all immune patients with cell mediated hypersensitivity to toxin. A partial response to lasting from 2 to 4 weeks was achieved in three of 13 nonimmune patients and in one of three nonimmune patients with cell-mediated hypersensitivity. Of 21 immune patients, nine had partial response lasting from 4 to 12 weeks. Of 13 immune patients with cell-mediated hypersensitivity, six had partial and five complete response lasting from 2 to 12 and from 1 to 25+ months, respectively. The overall response rate was 48%. The results suggest that diphtheria toxin may have a role in cancer immunotherapy. PMID- 6279292 TI - Radiotherapy of squamous, adeno- and large cell carcinoma of the lung. PMID- 6279293 TI - Mechanisms of promotion in nutritional carcinogenesis. PMID- 6279294 TI - Epidermal growth factor receptors interact with transforming growth factors produced by certain human tumor cells and are distinct from specific membrane receptors for phorbol and ingenol esters. PMID- 6279295 TI - Interaction of tumor promoters and growth factors with cultured cells. PMID- 6279296 TI - Inhibition of epidermal growth factor binding by phorbol esters, saccharin, and cyclamate and its implications in the mechanism of tumor promotion. PMID- 6279297 TI - A possible role of protein alkylation in phorbol ester action. PMID- 6279298 TI - Spin-labeled phorbol esters and their interaction with cellular membranes. AB - A series of homologous spin-labeled fatty acid analogs of the type 12-O-FASL (n,m)-phorbol-13-acetate [(n,m)PA] of 12-O-tetradecanoylphorbol-13-acetate (TPA) with variable chain length N of the fatty acid moiety and various positions of the nitroxide group were synthesized. Their irritant doses 50 (ID50) on the ear and their initiation-promoting activities on the back skin of NMRI mice were determined. The irritancy 1/ID50 of the (n,m)PA follows the simple linear relation ln(1/ID50) = alpha N + beta. In an "equilibrium" approach, the electron paramagnetic resonance (EPR) spectra measured in absence and in presence of erythrocytes (E) were used to determine the partition coefficients KM,0 between the erythrocyte membrane (M) and the extracellular solution (o). The KM,0 depend on the chain length N of the fatty acid moieties following the equation ln KM,0 = aN + b and show that the (n,m)PA are accumulated preferentially in the erythrocyte membrane. In a "kinetic" approach, the decay rate J(t)/J(o) of the EPR signal intensity of (n,m)PA molecules in absence and in presence of equilibrated E was measured during oxidation with ferricyanide and reduction with ascorbate, respectively. From the data obtained by the "kinetic" approach, a "two compartmental model" is shown to be the most simple approximation to describe the system, in which finite concentrations of the (n,m)PA are postulated for the extracellular and the membrane compartment only. EPR data indicate that generally the (n,m)PA are buried with their acyl chain in the E membrane with the phorbol moiety close to the outer membrane surface. Apparently only in case of (1,12)PA is the acyl chain long enough to be anchored in the lipid layer of the membrane in a way similar to the tetradecanoyl residue of TPA. A possible relationship with biological activities is discussed. PMID- 6279299 TI - Phorbol myristate acetate uncouples the relationship between beta-adrenergic receptors and adenyl cyclase in mouse epidermis: a phenotypic trait in papillomas. PMID- 6279300 TI - The role of free oxygen radicals in tumor promotion and carcinogenesis. PMID- 6279301 TI - Virus induction by tumor promoters. PMID- 6279303 TI - Cellular proliferation kinetics of the murine sarcoma induced by the Moloney sarcoma virus. AB - Cell proliferation kinetics of the sarcoma induced by Moloney virus was studied in newborn Swiss OF1 mice. After in vivo injection of tritiated thymidine, followed by autoradiography, it was shown that the majority of cells were actively proliferating (labelling index; 31%, growth fraction 78%). The mean cell cycle was 16 hr and cell loss was relatively low (cell loss factor 48%). The study of tumour specific activity with time after a single [in vivo] injection of [3H]dR or [125I]UdR did not demonstrate the same degree of cell loss as that calculated by autoradiography. This result is consistent with a massive reutilization of radioactivity released by normal tissues. PMID- 6279302 TI - Effect of tumor promoters in immunological systems--the macrophage as a target cell for the action of phorbol esters. PMID- 6279304 TI - Dibutyrylic cyclic AMP and theophylline inhibit proliferation of accessory cells in primary cultures of adrenomedullary cells. AB - Studies on isolated adrenal chromaffin cells in primary cultures may be seriously hampered by the presence of non-chromaffin, mainly fibroblast-like cells, which always occur in dissociates of adrenal medullary tissue and often outnumber the chromaffin cells by the end of the first week of culture, when no measures are taken to control their proliferation. The present study offers a new means to inhibit effectively the proliferation of these accessory cells by treating the cultures with dibutyrylic cyclic AMP (dbcAMP, 0.1 or 0.01 mM) and equimolar amounts of the phosphodiesterase inhibitor theophylline. With this treatment cultures of young rat adrenal chromaffin cells remain virtually free of accessory cells for two weeks of culture. Cultures of bovine adrenomedullary cells retain their initial amounts of non-chromaffin cells, which largely depends upon whether the primary cell suspensions have undergone differential plating prior to seeding. Suppression of accessory cell proliferation with dbcAMP and theophylline is partly due to maintaining differentiation of cortical cells, which otherwise dedifferentiate into rapidly dividing fibroblast-like elements. However, a more direct action of dbcAMP on accessory cells in terms of growth control is also conceivable. DbcAMP and theophylline in the doses applied do not impair the viability, ultrastructure and catecholamine-storing capacity of cultured chromaffin cells. PMID- 6279306 TI - Ultrastructural and cytochemical studies on the blood cells of the sea squirt, Ciona intestinalis. I. Stem cells and amoebocytes. AB - The ultrastructure and cytochemistry of the stem cell and amoebocytic blood cells of the sea squirt, Ciona intestinalis, are described. The stem cells are characterised by a high nuclear: cytoplasmic ratio and on undifferentiated cytoplasm. In thin sections, four types of amoebocytes can be demonstrated: non vacuolar and vacuolar hyaline amoebocytes, granular amoebocytes, and refractile amoebocytes. The cytoplasm of non-vacuolar amoebocytes contains many vesicles, which although structurally similar to primary lysosomes are in most cases acid phosphatase-negative. The granular amoebocytes enclose several microtubule containing granules and vacuoles with electron-dense vanadium deposits, while the refractive amoebocytes are characterised by their pleomorphic inclusions and spike-like pseudopodia. Both the granules and inclusions of these amoebocyte types are peroxidase- and acid phosphatase-negative indicating that they are probably not lysosomes. The possible interrelationships of the four amoebocyte types are discussed, and it is suggested that the vacuolar hyaline amoebocytes may differentiate into granular amoebocytes. PMID- 6279305 TI - Gastro-intestinal and neurohormonal peptides in the alimentary tract and cerebral complex of Ciona intestinalis (Ascidiaceae). AB - Polypeptide-hormone producing cells were localized in the alimentary tract and cerebral ganglion of Ciona intestinalis using cytochemical, immunocytochemical and electron-microscopical methods. Antisera to the following peptides of vertebrate type were employed: bombesin, human prolactin (hPRL), bovine pancreatic polypeptide (PP), porcine secretin, motilin, vasoactive intestinal polypeptide (VIP), beta-endorphin, leu-enkephalin, met-enkephalin, neurotensin, 5 hydroxytryptamin (5-HT), cholecystokinin (CCK), human growth (GH), ACTH, corticotropin-like intermediate lobe peptide (CLIP) and gastric inhibitory peptide (GIP). Immunoreactive cells were found both in the alimentary tract epithelium and in the cerebral ganglion for bombesin, PP, substance P, somatostatin, secretin and neurotensin. Additionally, in the cerebral ganglion only, there were cells immunoreactive for beta-endorphin, VIP, motilin and human prolactin. 5-HT positive cells, however, were restricted to the alimentary tract. No immunoreactivity was obtained either in the cerebral ganglion or in the alimentary tract with antibodies to leu-enkephalin, met-enkephalin, CCK, growth hormone, ACTH, CLIP and GIP. Prolactin-immunoreactive and pancreatic polypeptide immunoreactive cells were argyrophilic with the Grimelius' stain and were found in neighbouring positions in the cerebral ganglion. At the ultrastructural level five differently granulated cell types were distinguished in the cerebral ganglion. Granules were present in the perikarya as well as in axons. The possible functions of the peptides as neurohormones, neuroregulators and neuromodulators are discussed. PMID- 6279307 TI - Immunohistological study of the anterior pituitary gland - pars distalis and pars intermedia - in dwarf mice. AB - This immunohistological study of the anterior pituitary gland in Snell-Bagg dwarf mice demonstrates a total absence therein of somatotropin prolactin and thyrotropic cells. Gonadotropin, melanocorticotropic, LPH- and endorphin-positive cells are well developed; they appear to be hypertrophied and more numerous than in normal animals. PMID- 6279308 TI - Transmethylation reactions regulate affinity and functional activity of chemotactic factor receptors on macrophages. AB - Methylation mediated by S-adenosyl-L-methionine is required for the chemotaxis of mononuclear leukocytes. We investigated whether transmethylation reactions are required for normal functioning of chemotactic factor receptors. Three chemoatractant-mediated functions in macrophages, chemotaxis, the stimulated release of arachidonic acid from membrane phospholipids and superoxide production, are markedly depressed by agents that inhibit cellular methylation reactions. Treatment of macrophages with methylation inhibitors decreased the affinity of the N-formylated chemoattractant receptor present on these cells by a factor of 4.5, but did not significantly alter the total receptor on macrophages can exist in more than one affinity state and that an ongoing methylation reaction is required for the maintenance of the receptor in its higher affinity form. Inhibition of methylation lowers the affinity of the receptor and renders it nonfunctional or "uncoupled" in its ability to produce chemotaxis, superoxide and the release of a arachidonic acid from leukocyte membranes. PMID- 6279309 TI - Isolation and characterization of chicken DNA homologous to the two putative oncogenes of avian erythroblastosis virus. AB - The genome of avian erythroblastosis virus contains two independently expressed genetic loci (v-erbA and v-erbB) whose activities are probably responsible for oncogenesis by the virus. Both loci are closely related to nucleotide sequences found in the DNA and RNA of chickens and other vertebrates. We have isolated and characterized chicken DNA homologous to v-erbA and v-erbB. The two viral genes are represented by separate domains within chicken DNA (c-erbA and c-erbB), which are separated by a minimum of 12 kilobases (kb) of DNA and may not be linked at all. The nucleotide sequences shared by the viral and cellular erb loci are colinear, but the cellular loci are interrupted by multiple intervening sequences of various lengths. Polyribosomes prepared from normal chicken embryos contain two polyadenylated RNAs transcribed from c-erbA and two transcribed from c-erbB. The evident coding regions of these RNAs represent an unusually small fraction of the lengths of the RNAs, as if the 3' untranslated domains of the RNAs might be exceptionally large (3-11 kb). These findings indicate that the c-erb loci are normal vertebrate genes rather than genes of cryptic endogenous retroviruses, and that they may have a role in the metabolism of normal cells. It appears that the viral erb genes, like most other retrovirus oncogenes, have been copied from cellular genes. In the viral genome, the two genes are devoid of introns, but they remain independently expressed loci, and they remain colinear with the coding domains of their cellular progenitors. PMID- 6279310 TI - A symmetrical six-base-pair target site sequence determines Tn10 insertion specificity. AB - Transposon Tn10 inserts at many sites in the bacterial chromosome, but preferentially inserts at particular hotspots. We believe we have identified the target DNA signal responsible for this specificity. We have determined the DNA sequences of 11 Tn10 insertion sites and identified a particular 6 base pair (bp) symmetrical consensus sequence (GCTNAGC) common to those sites. The sequences at some sites differ from the consensus sequence but only in limited and well defined ways. The sequences at some sites differ from the consensus sequence than do sequences at other sites, and the consensus sequence and closely related sequences are generally absent from potential target regions where Tn10 is known not to insert. Other aspects of the target DNA can significantly influence the efficiency with which a particular target site sequence is used. The 6 bp consensus sequence is symmetrically located within the 9 bp target DNA sequence that is cleaved and duplicated during Tn10 insertion. This juxtaposition of recognition and cleavage sites plus the symmetry of the perfect consensus sequence suggest that the target DNA may be both recognized and cleaved by the symmetrically disposed subunits of a single protein, as suggested for type II restriction endonucleases. There is plausible homology between the consensus sequence and the very ends of Tn10, compatible with recognition of transposon ends and target DNA by the same protein. The sequences of actual insertion sites deviate from the perfect consensus sequence in a way which suggests that the 6 bp specificity determinant may be recognized through protein-DNA contacts along the major groove of the DNA double helix. PMID- 6279311 TI - Molecular organization of a Drosophila puff site that responds to ecdysone. AB - The 68C locus in the polytene chromosomes of Drosophila melanogaster salivary glands is puffed during the last half of the third larval instar and harbors the structural gene for sgs3, one of the glue polypeptides synthesized by the glands during this period. This puff regresses in response to the steroid hormone ecdysone. We have isolated a set of overlapping cloned segments that define approximately 50 kb of DNA at the 68C puff locus. Three polysomal poly(A)+ RNAs that are abundant in the salivary glands during the intermolt-puff stage are transcribed from three genes (II, III and IV) that map near the center of the cloned DNA in a 5 kb cluster region. These are the only transcripts from the 50 kb of 68C DNA detectable in these glands, and they are undetectable in other larval tissues at this stage, or in whole animals at other stages of development. Correlative criteria indicate that gene IV, which yields an RNA of 1.1 kb, is the structural gene for sgs3. Genes II and III, which yield RNAs of 0.36 kb and 0.32 kb, respectively, are oppositely oriented so that their promoters are adjacent, suggesting that this pair may form a single regulated unit, a suggestion that is enhanced by the fact that the pair is bounded by an inverted repeat of 0.3 kb elements. The possible, but as yet unidentified, functions of this gene pair are discussed. A 9.2 kb element belonging to a family of transposable elements called roo is inserted adjacent to the 5 kb cluster region in some but not other D. melanogaster strains. This insertion has no obvious effect on the transcription of genes II, III and IV. Although roo elements yield a 9 kb poly(A)+ RNA in embryos, no roo transcripts were detected in intermolt salivary glands, whether they do or do not contain an element at the 68C puff site. PMID- 6279312 TI - Uncoating of enveloped viruses. PMID- 6279313 TI - Two distinct intracellular pathways transport secretory and membrane glycoproteins to the surface of pituitary tumor cells. AB - The pituitary cell line, AtT-20, synthesizes adrenocorticotropic hormone (ACTH) as a glycoprotein precursor that is cleaved into mature hormones during packaging into secretory granules. The cells also produce an endogenous leukemia virus (MuLV) that is glycosylated after translation similar to the glycosylation of the ACTH precursor. Our evidence suggests that the envelope glycoprotein and some precursor ACTH get to the cell surface in a vesicle different from the mature ACTH secretory granule. Viral glycoproteins and ACTH precursor are released from the cells much sooner after synthesis than mature ACTH. Isolated secretory granules do not contain significant amounts of the envelope glycoprotein or ACTH precursor. Exposing cells to 8Br-cAMP stimulates release of mature ACTH four to five fold, but has little effect on the release of the ACTH precursor or the viral glycoproteins. We propose that the viral glycoproteins and some of the ACTH precursor are transported by a constitutive pathway, while mature ACTH is stored in secretory granules where its release is enhanced by stimulation. PMID- 6279314 TI - Evidence for a continual exchange of 5'-nucleotidase between the cell surface and cytoplasmic membranes in cultured rat fibroblasts. AB - Approximately 40% of the 5'-nucleotidase activity in cultured rat embryo fibroblasts was patent, as judged by enzymatic assays comparing the activity of intact cells with detergent-solubilized cells. The patent activity was inhibited when cells were incubated with anti-5'-nucleotidase serum at 2 degrees C, whereas latent activity (calculated as the difference between total and patent activity) was not. Latent activity was inhibited by antibody when the antiserum was added directly to detergent-solubilized cells or when cells were cultured in the presence of antiserum for several hours. Patent activity was inhibited by antibody, and cells were returned to culture in antibody-free medium; after 12 hr, 30% of the total activity was expressed in intact cells and 60% of the anti 5'-nucleotidase, assayed by the binding of sheep antirabbit antibodies to intact cells, was lost from the cell surface, indicating an exchange of 5'-nucleotidase between the latent and patent compartments. Cytochemical studies showed that the patent activity was located on the cell surface and that latent activity was present in cytoplasmic vacuoles and vesicles, and in the Golgi complex. Over 30% of the anti-5'-nucleotidase internalized during 6 hr in culture returned to the cell surface after a further 9 hr, indicating a continual exchange of the enzyme between the cell surface and cytoplasmic membranes. PMID- 6279315 TI - Decreased uptake and retention of rhodamine 123 by mitochondria in feline sarcoma virus-transformed mink cells. AB - A reduce uptake and retention of the mitochondria-specific membrane potential probe rhodamine 123 by feline sarcoma virus (FeSV)-transformed mink fibroblasts (64F3) has been detected. The decreased accumulation of rhodamine 123 by 64F3 mitochondria is not due to abnormal plasma membrane dye permeability, since after microinjection of the dye these cells are still unable to retain the dye at levels comparable to the untransformed parental cells, CCL 64. Nigericin, an ionophore that mediates an electrically neutral exchange of protons for potassium ions resulting the elimination of the pH gradient across the mitochondrial membrane and a compensatory increase in mitochondrial membrane potential with continued respiration, increases both the dye uptake and the retention time in transformed 64F3 cells. These results suggest that mitochondria in FeSV transformed mink cells may have an abnormally low mitochondrial membrane potential accompanied by a relatively high pH gradient. Since anioic metabolites such as pyruvate and glutamate are accumulated by mitochondria in proportion to the delta pH across the mitochondrial membrane, the abnormal mitochondria described here may contribute to the abnormal metabolic state of FeSV-transformed cells. PMID- 6279316 TI - Expression of neuronal markers in chick and quail embryo neuroretina cultures infected with Rous sarcoma virus. AB - Cultures of neuroretina (NR) cells from 7-day chick and quail embryos were infected with ts NY-68, a thermosensitive mutant of Rous sarcoma virus (RSV) which transformed NR cells at 36 degrees C. The following differentiation markers for neurones were studied: tetanus toxin-binding sites at the cell surfaces, presence of synapses, and the specific activity of the enzymes choline acetyltransferase (CAT) and glutamic acid decarboxylase (GAD). Appearance of synapses and expression of CAT were similar in control and transformed cultures. Tetanus toxin-binding cells were observed in transformed primary cultures and also in quail NR subcultures. GAD-specific activity was markedly stimulated in chick and quail primary cultures transformed by ts NY-68 and further increased in subcultures of ts NY-68-transformed quail NR cells. Stimulation of GAD activity is controlled by the transforming (src) gene of RSV since it was not observed in cultures infected with RAV-1, a leukosis virus which lacks the src gene. These data show that infection of chick and quail NR cultures with RSV results in the transformation of cells with neuronal markers. PMID- 6279317 TI - Changes in endonuclease activity and in chromatin structure of rat hepatocytes during fetal and neonatal life. AB - A developmental study of rat hepatic endonuclease has been performed. Nuclei, from different stages of hepatocyte maturation, were analyzed for endogenous endonuclease activity. The chromatin extracted from these nuclei does not show any fragmentation during the first 17 days of fetal development. On the 18th day of fetal life there is a massive increase in specific endonuclease activity. At birth this activity reaches a maximum level (3.5 units/mg DNA); thereafter it undergoes a gradual decrease. The size of the basic DNA repeats produced by the endonuclease action is 218.9 +/- 1.6 in 18-day-old fetuses and decreases to 204.9 +/- 2.5 in 19-day-old fetuses, a value which remains constant in the following fetal and postnatal life. This difference in monomer size is due to changes in the chromatin structure. Micrococcal nuclease digests show that the "nucleosome core" does not change during hepatocyte development. Therefore, the difference in size of the endonuclease DNA fragments must be due to the linker regions. PMID- 6279318 TI - The migration of lymphocytes across specialized vascular endothelium. IV. Prednisolone acts at several points on the recirculation pathways of lymphocytes. PMID- 6279319 TI - [Immunoenzymatic detection of IgA and IgG antibodies to HSV-1, HSV-2 in sera of patients with cervical cancer (author's transl)]. PMID- 6279320 TI - [Studies on abnormal hemoglobins in China: structural analysis of a case of Hbj Calabria (author's transl)]. PMID- 6279321 TI - [A simplified method for preparation of restriction endonuclease BamHI (author's transl)]. PMID- 6279322 TI - A system of nomenclature for poliovirus strains: a WHO memorandum. AB - As a result of recent developments in the biological and biochemical characterization of polioviruses, intratypic differentiation of poliovirus strains has become a powerful epidemiological tool. However, use of this tool should be linked with a better system of nomenclature.It is recommended that future poliovirus isolates should be identified by type, country (or city), strain number, and year of isolation. The system should be applied universally from the time of publication of this Memorandum. PMID- 6279323 TI - A serological study of cytomegalovirus and herpes simplex virus infections in Peninsular Malaysia. AB - Healthy Malaysians from various parts of Peninsular Malaysia were examined for CF antibodies against cytomegalovirus (CMV) and herpes simplex virus (HSV) type 2. CMV antibodies were detected in 1114 out of 1556 persons (71.6%) and HSV antibodies were detected in 954 persons out of 1554 (61.4%). The age distribution patterns were similar for the two infections, with maximum prevalence at 5 - 14 years of age. Prevalence was higher in women than in men. There were no significant differences among the Malay, Chinese, and Indian groups of the population with respect to CMV, 72 - 78% possessing antibodies, but in the case of HSV, 76% of the Chinese had antibodies, compared with 57 - 60% of the Malays and Indians. More than 90% of newborn infants had CMV and HSV CF antibodies, confirming the highly immune status of childbearing women in Malaysia. No CMV specific IgM was detected in the Malaysian neonates examined but this does not exclude the possibility of congenital infection. PMID- 6279324 TI - DNA repair in normal human and xeroderma pigmentosum group A fibroblasts following treatment with various methanesulfonates and the demonstration of a long-patch (u.v.-like) repair component. AB - Excision repair of DNA in normal and xeroderma pigmentosum complementation group A fibroblasts were examined following treatment with methyl-, ethyl-, and isopropyl methanesulfonate. Studies utilizing repair synthesis methods and inhibition with arabinofuranosyl cytosine revealed two distinct phases of repair; one commencing and terminating within the first 3-5 h after the treatment, and a second much longer phase extending from 9-35 h post-treatment. Both phases of repair have a long-patch (u.v.-like) component, which establishes for the first time the existence of this mode of repair in response to alkane sulfonate damage. While xeroderma cells display somewhat fewer alkaline labile sites in their DNA following alkylation treatment than do their normal counterparts (consistent with earlier observations suggesting a deficiency in a glycosylase or apurinic endonuclease), we are unable to demonstrate a deficiency of these cells in either of the two phases of repair. PMID- 6279325 TI - Correlation between retinoid inhibition of N-methyl-N-nitrosourea-induced mammary carcinogenesis and levels of retinoic acid binding proteins. AB - A correlation was made between the ability of retinoids to suppress N-methyl-N nitrosourea (MNU)-induced mammary carcinogenesis and the levels of cytosolic retinoic acid binding proteins (cRABP) in the cytosol of MNU-induced mammary tumors. Although retinyl acetate and N-(4-hydroxyphenyl)-retinamide were found to be effective inhibitors of mammary carcinogenesis in intact hosts, both retinoids were significantly more active in ovariectomized rats than in intact animals. Quantitative analysis of cRABP in the tumors indicated that mammary cancers arising in animals which were ovariectomized one week after MNU administration contained significantly increased concentrations of cRABP compared to cancers appearing in intact rats. In addition, when animals bearing palpable mammary tumors were ovariectomized, the tumors which continued to grow contained significantly higher levels of cRABP than did tumors which stopped growing or regressed. These data suggest that the selective inhibition of ovarian hormone independent mammary cancer by retinyl acetate and N-(4-hydroxyphenyl)retinamide may be mediated through an increased level of cRABP in tumor cells of ovariectomized hosts. PMID- 6279326 TI - Different patterns of benzo[a]pyrene metabolism of purified cytochromes P-450 from methylcholanthrene, beta-naphthoflavone and phenobarbital treated rats. AB - An improved high-pressure liquid chromatography system was used to analyze the amount of benzo[a]pyrene metabolites formed in reconstituted microsomal mixed function oxidase systems containing different cytochromes P-450. We separated twelve identified and seven unknown metabolites of BP which included three diols: the 9,10-, 4,5-and 7,8-dihydrodiols; four phenols, 9-,7-, 1-, and 3 hydroxybenzo[a]pyrene (OH-BP); and three quinones: the 1,6-. 3,6-, and 6,12 quinones. Two additional peaks co-migrated with synthetic 4-OH-BP and 5-OH-BP, respectively. The former, designated fraction 1, was shown by u.v. spectra to contain primarily the 4,5-epoxide with small amounts of 4-OH-BP. The total metabolism of BP was found to be approximately 20-fold greater with the cytochrome P-450 from the 3-methylcholanthrene (P-450 3-MC) and beta naphthoflavone (P-450 BNF) treated rats than with the phenobarbital preinduced cytochrome P-450 (P-450 BP). 3-OH-BP ad 9-OH-BP were the major phenolic products for both P-450 3-MC and P-450 BNF whereas the 3-OH-BP and 1-OH-BP were the major phenolic products for P-450 BP. The ratio of total phenols to diols was found to be 3.34, 4.85 and 0.70 for P-450 3-MC, P-450 BNF and P-450 PB. The major dihydrodiol generated by P-450 3-MC and P-450 BNF was 7,8-dihydroxy-7,8 dihydrobenzo[a]pyrene, whereas the 9,10-diol was the major diol from P-450 PB. The amount of 1,6- and 3,6-quinones produced was greater than the 6,12-quinone with the P-450 3-MC and P-450 BNF but all three quinones were produced in low and equal amounts by the P-450 PB. In respect to the percent metabolites formed at a given region of the BP, P-450 3-MC and P-450 BNF preferred oxidation at the 1, 3 positions, 6 position and the 7, 8 positions, whereas the P-450 PB preferred oxidation at the 4, 5 position. This study demonstrates the unique positional specificity of different forms of cytochrome P-450 which may regulate the balance between activation and detoxification pathways of polycyclic aromatic hydrocarbon metabolism. PMID- 6279327 TI - Protease inhibitors from processed legumes effectively inhibit superoxide generation in response to TPA. AB - Crude extracts containing protease inhibitors from edible legumes (canned chick peas, canned kidney beans and bean curd) were capable of blocking the superoxide response to human polymorphonuclear leukocytes produced by the tumor promoter 12 O-tetradecanoyl-phorbol-13-acetate (TPA). Protease inhibitors purified from crude extracts more effectively blocked the superoxide response produced by TPA. Bowman Birk soybean inhibitor was more effective in blocking this effect of the tumor promoter than Kunitz soybean inhibitor. The significance of protease inhibitors in edible legumes and the possible role of free oxygen radicals in tumor promotion are discussed. PMID- 6279328 TI - Inhibition of metabolic cooperation in Chinese hamster V79 cells in culture by various polybrominated biphenyl (PBB) congeners. AB - Using an in vitro assay system to measure a form of cell-cell communication in Chinese hamster V79 cells, several purified polybrominated biphenyl (PBB) congeners were analyzed for their ability to affect colony-forming ability and "metabolic cooperation" in these cells. Results clearly demonstrate the assay's ability to distinguish differences in the cytotoxic and metabolic cooperation inhibiting properties of these congeners related to their structure. It appears that if a carbon ortho to the bridge carbon of each phenyl ring is brominated, the molecule has the property by which metabolic cooperation is inhibited. The results suggest that several PBB congeners share at least one property associated with several known tumor promoters, such as the phorbol esters. Although we have not ruled out that the mixture of PBB congeners might contain initiators or that various PBB metabolites would be mutagen/initiators, based on these and other results, it was concluded that the reported carcinogenic potential of PBB seems to be mediated by its action as a tumor promoter, rather than as a complete carcinogen or an initiator. PMID- 6279329 TI - Metabolism and DNA binding of benzo[a]pyrene in cultured human bladder and bronchus. AB - The metabolism of benzo[a]pyrene (BP) was examined in explant cultures of human bladder and bronchus. Three-day cultures were exposed to radiolabeled BP for 24 h, and the metabolism was determined by analysis of the level of binding of reactive metabolites to DNA, and by the release of metabolites into the medium. For a given individual, the DNA binding level and extent of metabolism was usually higher in the bladder than in the bronchus. In specimens obtained from 16 individuals, the average DNA-binding levels for BP-DNA adducts following a 24 h exposure to 1 microM BP were 6.4 +/- 5.0 mumol BP/mol deoxyribonucleotide for the bladder and 3.1 +/- 1.9 mumol BP/mol deoxyribonucleotide for the bronchus. The major BP-DNA adduct in both tissues co-chromatographed with one of the adducts formed by reaction of r-7, t-8-dihydroxy-t-9,10-oxy-7,8,9,10 tetrahydrobenzo[a]pyrene with deoxyguanosine using high-pressure liquid chromatography. In tissues obtained from the same individual, the binding levels of BP metabolites to bladder cell DNA was not strongly correlated to that of bronchial cell DNA (r = 0.55). The medium of both tissues contained small amounts of free, unconjugated metabolites of BP (less than 3% of the total) and large amounts (30-86% of the total) of unidentified, highly polar material. Human bladder appears to be the most active explant tissue yet studied with respect to its ability to activate BP to DNA binding forms. The relevance of this observation to human bladder cancer is, as yet, unknown. PMID- 6279331 TI - Hepatic arterial and portal flow in cardiogenic and hemorrhagic shock in awake dogs. AB - The changes in liver blood flow produced by experimental cardiogenic and hemorrhagic shock are relatively unexplored. Fifteen unanesthetized dogs in which electromagnetic blood flow transducers had been implanted on the common hepatic artery and portal vein were subjected to acute myocardial infarction by mercury embolization of the circumflex coronary artery. Another group of ten dogs were bled to an arterial pressure of 40 mmHg and maintained at that level for 2 hours before reinfusion. Six additional dogs in which blood flow transducers had been implanted on the superior mesenteric artery and portal vein also were subjected to hemorrhage. In three of the six, phenoxybenzamine was infused directly into the superior mesenteric artery 45 minutes prior to bleed-out. During cardiogenic shock, both hepatic arterial and protal venous flow fell. However, whereas protal flow continued to fall, eventually stabilizing at values 36 +/- 3% of control, hepatic arterial flow subsequently rose, reaching values 93 +/- 9% above control. Total liver blood flow, after an initial decline to 53 +/- 4% of control levels rose as a result of the increased hepatic arterial flow to 73 +/- 4% of control values. During hemorrhage, both hepatic arterial and protal venous flows decreased as aortic pressure fell. Within 5 minutes of reinfusion, hepatic arterial flow surpassed its control values. Portal flow also increased but, on a percentage basis, not to so great an extent. Flow in hepatic artery remained high for 40 minutes after reinfusion, whereas portal flow rapidly decreased to levels seen at the end of hemorrhage. In hemorrhage without alpha-adrenergic receptor blockade the superior mesenteric bed constricted, thereby supporting systemic pressure. With alpha-adrenergic blockade, however, mesenteric flow became pressure-dependent and no longer acted as a homeostatic mechanism. PMID- 6279332 TI - Angiotensin II and alpha-adrenergic control of the intrarenal circulation in hemorrhage. AB - Alpha receptor and angiotensin II blockade were used to study renal cortical vasoconstrictor mechanisms during hemorrhage to 70 mmHg in chloralose-urethane anesthetized dogs. A freeze-dissection 133Xe disappearance technique was utilized to assess renal blood flow patterns. Hemorrhage alone caused a 30% decrease in total renal blood flow (TRBF) and a 50% decrease in outer cortical blood flow. Inner cortical flow decreased approximately 30%. Outer medullary blood flow decreased 25%. Renal arterial infusion of phentolamine beginning 20 min posthemorrhage produced no alteration in the expected posthemorrhage TRBF or its distribution. Plasma renin activity increased 6-7 fold in the hemorrhage group as well as in the hemorrhage with phentolamine group. Saralasin treatment beginning 20 min posthemorrhage produced a pattern in which neither TRBF nor cortical blood flow was significantly reduced by the hemorrhage. It therefore appears that angiotensin II is a renal cortical vasoconstrictor during hemorrhage. These data demonstrate that during the first 30 minutes of hemorrhage the renal cortical vasoconstriction is mediated by angiotensin II and appears to be independent of alpha-adrenergic mechanisms. PMID- 6279330 TI - The metabolism and activation of polycyclic aromatic hydrocarbons in epithelial cell aggregates and fibroblasts prepared from rat mammary tissue. AB - The metabolism of 7,12-dimethylbenz[a]anthracene (DMBA), benzo[a]pyrene (BP) and benz[a]anthracene (BA) by epithelial cell aggregates and fibroblasts in culture has been investigated using mammary tissue obtained from female Wistar rats. The results show that: (a) both types of mammary cells metabolise all three hydrocarbons into ether- and water-soluble derivatives; (b) the patterns of metabolites produced by epithelial cells and fibroblasts are similar but that fibroblasts form more of the water-soluble materials; (c) the major dihydrodiols formed are the 8,9-dihydrodiols of BA and DMBA and the 7,8- and 9,10-dihydrodiols of BP; (d) all three hydrocarbons are metabolise to form products that bind covalently to protein but only the potent carcinogens BP and DMBA are metabolised to form derivatives that react covalently with DNA; and (e) the chromatographic profiles of the hydrocarbon-deoxyribonucleoside adducts formed in epithelial cells treated with either BP or DMBA are similar to those obtained in analogous experiment with fibroblasts. PMID- 6279333 TI - Sodium, divalent cations, and guanine nucleotides regulate the affinity of the rat mesenteric artery angiotensin II receptor. PMID- 6279334 TI - Alpha subunit of the glycoprotein hormones: secretion by human malignancies. PMID- 6279335 TI - A one-step determination of serum 5'-nucleotidase using a centrifugal analyzer. AB - We describe a one-step kinetic method for the determination of 5'-nucleotidase (EC 3.1.3.5). Inosine is formed by the hydrolysis of inosine 5'-monophosphate which is catalyzed by seric 5'-nucleotidase, and then is converted to hypoxanthine by nucleoside phosphorylase. Two moles of hydrogen peroxide are formed for each mole of hypoxanthine oxidized to urate by xanthine oxidase. The rate formation of hydrogen peroxide is monitored at 510 nm using the oxidation of the chromogenic system 3,5-dichloro-2-hydroxybenzenesulfonic acid/4 aminophenazone in the presence of peroxidase. beta-Glycerophosphate inhibits the unspecific cleavage of the substrate by alkaline phosphatases. Inorganic phosphate is added to improve the reagent stability, and ferrocyanide to reduce bilirubin interference. Automation of the technique requiring 20 microliter of serum on a centrifugal analyzer is also described. PMID- 6279336 TI - In vitro studies on calcium activated phosphatidylinositol phosphodiesterase of erythrocyte ghosts from normal individuals and those with myotonic muscular dystrophy. AB - Myotonic muscular dystrophy is an inherited disorder affecting many organs, though the underlying biochemical defect is unknown. A recent publication [1] suggested that the metabolic lesion may be associated with defective phospholipid metabolism. These workers observed impaired calcium-stimulated phosphatidic acid accumulation in red cell ghosts from individuals with myotonic dystrophy compared with normal controls. The present study investigated some points of calcium activated phosphatidylinositol metabolism in red cell ghosts from patients with myotonic dystrophy, those at risk of developing the disease and normal individuals. No differences between the three groups could be found in the incorporation of 32P into endogenous phosphatidylinositol nor in the distribution of label between the various phosphatidylinositols. Additionally, no differences were observed in either basal or calcium-activated phosphatidylinositol phosphate breakdown by phosphodiesterase. This would suggest that the observed decreased phosphatidate accumulation [1] may be due to impaired diacylglycerol kinase activity. PMID- 6279337 TI - Adrenocortical function in children with precocious sexual development during treatment with cyproterone acetate. AB - Adrenal function was studied in thirty-two children with precocious sexual development who were being treated with cyproterone acetate (CPA) at doses ranging from 68 to 175 mg. m2. day for periods lasting from 2 to 79 months. In eighteen children the adrenocortical function evaluation was made before and during CPA treatment. In these eighteen patients, the mean basal plasma cortisol level during the morning hours was 11.2 +/- 4.6 micrograms/dl (m +/- SD) before treatment and fell significantly to 7.2 +/- 4.1 micrograms/dl (P less than 0.02) during therapy. In fifteen patients tested during insulin hypoglycaemia the cortisol peak fell from 21.6 +/- 5.5 micrograms/dl before treatment to 16.7 +/- 6.8 micrograms/dl (P less than 0.05) during CPA therapy. There was a significant inverse correlation between this peak and the dose of CPA but no correlation was found between the cortisol response and duration of treatment. In eight of twenty patients tested, urinary free cortisol levels were undetectable during treatment. No change in basal plasma ACTH levels were demonstrated using standard radioimmunoassay techniques. In the patient receiving the highest dose of CPA and showing complete suppression of the adrenal axis, prolonged stimulation with ACTH Depot demonstrated a responsive adrenal gland. Addition of a replacement dose of cortisol to the CPA treatment led to the rapid development of the typical signs of Cushing's syndrome. It was concluded that despite the evidence of adrenal suppression by CPA, cortisol supplementation is not necessary and may not even be contraindicated. PMID- 6279338 TI - Human hair follicles, a convenient tissue for genetic studies on carcinogen metabolism. AB - Basal levels of benzo(a)pyrene metabolism were measured in hair follicles of seven monozygotic twins, eight dizygotic twins and ten pairs of unrelated individuals. Organic soluble metabolites were separated by thin-layer chromatography, visualised by autoradiography and quantified by scanning of the films. Activity was expressed as pmol 7,8- and 9,10-dihydrodiol metabolites of benzo(a)pyrene per micrograms DNA per hour. Intra-twin differences in benzo(a)pyrene metabolism for monozygotic twins were smaller than for dizygotic twins and intra-pair differences for dizygotic twins were smaller for pairs of unrelated individuals. The results clearly suggest that individual differences in benzo(a)pyrene metabolism in hair follicles are partly genetically determined. Thus, hair follicles may be used for investigations on the suggested relations between genetic predisposition to carcinogen-induced cancer and individual differences in carcinogen metabolism. The relevance of these findings to research into the induction of neoplasms by carcinogens in epithelial tissues is discussed. PMID- 6279339 TI - Pseudodeficiency of alpha-galactosidase A. AB - Apparent deficiency of alpha-galactosidase A was observed in a 51-year-old, clinically healthy male, with no clinical symptoms of Fabry disease, and without excess urinary excretion of ceramide trihexoside. The deficiency, which was similar to that found in Fabry disease patients, could be demonstrated using both synthetic and natural substrates. This pseudodeficiency was transmitted in his family by classical X-linked inheritance. His wife showed enzyme activity in the normal range, two daughters were heterozygotes for this mutation as demonstrated by hair root assay, and three sons showed normal alpha-galactosidase activity. Kinetic studies in cultured skin fibroblasts indicated a five-fold increase in the apparent Km and a greater heat stability of the residual alpha-galactosidase activity when compared to controls. These data indicate that the residual enzyme activity in this mutation behaves similarly to that observed in Fabry disease patients but does not cause any clinical abnormalities. PMID- 6279340 TI - Present nosology of the Cenani-Lenz type of syndactyly. AB - Synostoses of the carpals and metacarpals with oligodactyly were noted in a man whose brother was similarly affected. Since the proband's two children are normal, autosomal recessive transmission is probable. Although abnormalities of the feet, and particularly radioulnar synostosis, are lacking, this malformation is comparable to the Cenani-Lenz type of syndactyly. Interfamilial variations of published cases suggest subdivision of a new nosologic group. PMID- 6279341 TI - Specific in vitro antibody response to varicella zoster. AB - Peripheral blood mononuclear cells from normal donors were stimulated in vitro by varicella zoster (VZ) antigen and antibody measured by an ELISA technique. Antibody was produced in 14 of 19 individuals. This antibody was specific for VZ with no binding to herpes simplex virus (HSV), even when the mononuclear cells were taken from individuals who had circulating antibody to VZ and HSV. The failure of antibody production in mononuclear cell cultures from some normals, all of whom had serum antibody, was analysed using reciprocal combinations of E+ and E- cells from producers and non-producers. E+ cells from producers could help E- producer cells make antibody, but E+ cells from non-producers could not help E cells from non-producers to do so. This result suggests a deficiency in circulating E- cells in non-producers. Four of four patients with untreated Hodgkin's disease were non-producers using blood mononuclear cell cultures, but their spleen cells produced antibody readily. This suggests that failure of in vitro antibody production in immune individuals may be due to sequestration of responsive B cells in spleen and other sites. This technique allows further study of the mechanisms of immunity to VZ in normal and immune-compromised individuals. PMID- 6279342 TI - The isolation of the antibody moieties of immune complexes from serum by the pepsin digestion of conglutinin-anti-conglutinin complexes. AB - A technique is described which allows the antibodies of circulating immune complexes to be isolated as their F(ab')2 fragments. The method is based on the precipitation of the complexes by the sequential addition of conglutinin and anti conglutinin, and the subsequent digestion of these precipitates by pepsin. Using this technique it has been possible to show antibodies to Epstein-Barr (EB) virus antigens in the immune complexes of patients with Burkitt's lymphoma and to microbial antigens in two patients with nephritis. By substituting DNAase for pepsin it has also been possible to show antibodies to DNA-containing nuclear antigens in the serum of patients with systemic lupus erythematosus. PMID- 6279344 TI - Cytotoxicity of human peripheral blood monocytes against Trichomonas vaginalis. AB - Human peripheral blood monocytes were purified by adherence on microexudate coated plastic and detached by exposure to 1 mM ethylenediamine tetracetic acid. Lysis of Trichomonas vaginalis was measured as release of 3H-thymidine from prelabelled protozoa after 48 hr of incubation. Human blood monocytes had appreciable levels of spontaneous cytotoxicity against Trichomonas vaginalis at effector-to-target cell ratios ranging from 3:1 to 25:1. Cytotoxicity was expressed with fetal bovine serum and human AB serum. Unseparated mononuclear cells, containing 10-25% monocytes as assessed by staining with non-specific esterase, were significantly cytotoxic against Trichomonas vaginalis but cytolysis levels were lower than those of purified monocytes. Lymphoid cells depleted of adherent cells by repeated incubations on plastic (less than 2% esterase-positive cells) had little cytotoxicity against these protozoa. In vitro exposure to silica natural 40-75% inhibition of the cytotoxic capacity of monocytic preparations. The natural cytotoxic capacity of human monocytes against Trichomonas vaginalis could represent one line of resistance against these protozoa. PMID- 6279343 TI - Studies on the kinetic properties and subcellular localization of adenosine diphosphatase activity in human peripheral blood lymphocytes. AB - Using a recently developed radioassay, the conditions for measuring adenosine diphosphatase (ADPase) activity in human lymphocytes were optimized. Kinetic studies showed that the activity was optimal at pH 8.0 and required 0.2 mM MgCl2. The Km of the enzyme for ADP was 0.03 mM. Analytical subcellular fractionation showed that the ADPase activity was distributed between the plasma membrane and mitochondria. Studies with the non-penetrating inhibitor, diazotized sulphanilic acid, indicate that plasma membrane ADPase activity is located on the external aspect of the cell. Polymorphonuclear leucocytes were found to have about three times the ADPase activity. Lymphocytes, therefore, have the ability to generate AMP on their surface which can be further metabolized to adenosine by ecto-5' nucleotidase. A role for this system is discussed. PMID- 6279345 TI - Nephrotic syndrome in association with a trophoblastic neoplasm. PMID- 6279346 TI - The effect of treatment on growth and deformity in hypophosphatemic vitamin D resistant rickets. AB - Vitamin D-resistant rickets is characterized by short stature, lower extremity deformities, and defective mineralization of bone. While basic biochemical defects vary among involved individuals, all patients show a failure of the proximal tubule of the kidney to resorb inorganic phosphate. Laboratory findings consist of low serum phosphorus, elevated alkaline phosphatase, and abnormal serum calcium. Previously recommended treatment programs of high doses of vitamin D have effected some roentgenographic improvement in the rachitic lesions, but no related increase in height or severity of deformities has been associated with significant complications resulting from vitamin D toxicity. Daily administration of low doses of vitamin D and oral phosphates has more recently been suggested to be beneficial in promoting growth and preventing deformities. Thirteen children with documented vitamin D-resistant rickets were treated with oral phosphate and low doses of vitamin D for a mean of five years, and followed for a mean of ten years. Partial roentgenographic resolution of rachitic lesions was similar to those who received vitamin D alone. While the majority had consistently lowered alkaline phosphatases, no patient had consistently normal phosphate levels. No patient exceeded the third percentile in height. One half required osteotomies of the lower extremities. All osteotomies (eight) performed before maturity required revision, whereas those done after maturity (12) did not. Since no apparent clinical roentgenographic benefit can be documented by the addition of oral phosphate to low doses of vitamin D, we would not recommend continually doing so at this time. PMID- 6279347 TI - Peripheral nerve electrical stimulation. PMID- 6279348 TI - The natural history of metastatic synovial sarcoma: experience of the Southwest Oncology group. AB - Of 35 treated cases of metastatic synovial sarcoma, there was a slight male predominance (60%). The median age was 33 years, with 88% of the patients being white. The lower extremity was the most common primary site, occurring in 66% of the cases. The most common metastatic involvement was pulmonary (33 patients), with only three patients having lymph node involvement. The low incidence of lymph node involvement and the poor prognosis when positive lymph nodes are found suggest that prophylactic lymph node resection is unwarranted. The median time from evidence of metastases to death was ten months. The complete and partial response rate to chemotherapy in this series was 50%; however, duration of the response was rather short with recurrence of metastatic disease a median of seven months after response was first noted. The size of the primary lesion was greater than 5 cm in all but one of the cases. The local recurrence rate was 70% in those cases undergoing local resection and 30% in those cases in which local resection plus irradiation to the primary lesion was undertaken. No case in which radical resection of the primary tumor was undertaken had a local recurrence. PMID- 6279349 TI - The classics. Checkrein shoulder: a type of "frozen" shoulder: diagnosis and treatment by manipulation and ACTH or cortisone by T. B. Quigley, M.D. PMID- 6279350 TI - Radiation-induced sarcoma of bone: clinical and radiographic findings in 43 patients irradiated for soft tissue neoplasms. AB - Sarcomas rarely follow irradiation of benign lesions of bone as the dangers of this form of therapy is well recognised. Forty-three patients with soft tissue neoplasms - mainly carcinoma - were irradiated and developed sarcomas in the radiation field. Twenty-four of these patients were seen at this centre in the last decade, a considerable increase compared to earlier years. Radiation osteitis may indicate the nature of the tumour but was only present in 50%. A purely lytic or purely sclerotic pattern was commoner than a mixed lytic and sclerotic appearance. CT scan, bone scan and angiography were helpful in delineating disease but did not assist in diagnosing the type of tumour. A tumour arising within the irradiated field is an indication for biopsy. Prognosis of radiation-induced sarcoma (RIS) remains poor, only five of the 43 patients survived 5 years. PMID- 6279351 TI - Circadian variation in adrenergic responses in asthmatic subjects. AB - 1. To determine whether circadian variations in adrenergic responsiveness might underlie nocturnal wheezing in asthma, we measured cardiovascular, airway and plasma adenosine 3':5'-cyclic monophosphate (cyclic AMP) responses to stepwise infusions of L-adrenaline (0.01, 0.03 and 0.075 microgram min-1 kg-1) at 4 h intervals over 24 h in five extrinsic asthmatic men. 2. Peak expiratory flow, blood pressure, heart rate and plasma cyclic AMP showed a significant circadian variation with peak values at 16.00 hours and trough values at 04.00 hours. 3. The beta 2-adrenoceptor-mediated increases in peak flow and cyclic AMP were similar at all times, but adrenergic responsiveness (measured by response/log dose of infused adrenaline) was greater at 04.00 hours than at 16.00 hours because of the lower baseline values at night. 4. Blood pressure and heart rate responses to adrenaline infusions did not significantly differ over 24 h. 5. Airway responses to inhaled adrenaline were studied on the second day; the mean peak flow after adrenaline was similar at 16.00 hours to that at 04.00 hours and since the pretreatment values were lower at 04.00 hours, the magnitude of response to inhaled adrenaline was greater at night. 6. We conclude that there is no significant circadian change in adrenergic responses in asthma and that adrenoreceptor dysfunction is not important in the pathogenesis of nocturnal asthma. PMID- 6279352 TI - Reversible resistance to the renal action of parathyroid hormone in human vitamin D deficiency. AB - 1. The response to exogenous parathyroid hormone (PTH) was tested in normal subjects and patients with osteomalacia due to vitamin D deficiency; 200 MRC units of bovine PTH were administered intravenously. 2. The rise in plasma adenosine 3':5'-cyclic monophosphate (cyclic AMP) and the increase in urinary excretion of cyclic AMP were reduced in the patients with vitamin D deficiency. After treatment with vitamin D the responses returned to normal. 3. It is suggested that this reversible resistance is due to the secondary hyperparathyroidism associated with vitamin D deficiency. PMID- 6279353 TI - Heritability of the low density lipoprotein receptor activity of human blood mononuclear cells: studies in normolipidaemic adult male twins. AB - 1. The concordance of low density lipoprotein (LDL) receptor activity of blood mononuclear cells was examined in 26 pairs of monozygotic and 17 pairs of dizygotic normolipidaemic young adult male twins. 2. Receptor activity was quantified as the degradation of 125I-labelled LDL during a 6 h incubation, after derepression of the cells for 72 h in lipoprotein-deficient medium. 3. The total variance of receptor activity was similar in the two groups of twins. In contrast, within-pair variance was five times greater in dizygotic twins than in monozygotic twins (P less than 0.001). 4. Estimates of heritability, mostly based on the intra-class correlation coefficients (monozygotic, r = 0.83; dizygotic, r = 0.39), ranged from 0.72 to 1.05. 5. These results suggest that the maximal LDL receptor activity of peripheral cells in normolipidaemic subjects is largely genetically determined. PMID- 6279354 TI - The new cephalosporins and penicillins. PMID- 6279355 TI - Differences of adenosine kinases from various mammalian tissues. AB - 1. Adenosine kinases purified to homogeneity from various mammalian tissues have a monomeric structure, and their molecular weights were estimated around 40,000. 2. The enzyme activity per wet weight of tissue appears to vary from source to source, but the specific activities of the final enzyme preparations were alike, which were 6.3--7.8 mumol/min/mg protein. 3. No or small difference was observed in the kinetic properties among all seven adenosine kinases. 4. Some differences in structure were observed among five liver enzymes from human, rabbit, rat, mouse and Mongolian gerbil, but no difference was observed between the enzymes from human placenta and liver, or the enzymes from rat liver and brain. PMID- 6279357 TI - Allergy to zinc dimethyldithiocarbamate in rubber gloves. PMID- 6279356 TI - Biological properties of a calcium-modulator protein from Phycomyces blakesleeanus. AB - 1. A modulator protein having properties similar to calmodulin-like proteins could be detected in Phycomyces the first day of growth. 2. The modulator protein from Phycomyces was partially purified to a specific activity of 9,000 Units/mg protein. 3. The modulator protein was similar to calmodulin with respect to: (a) the ability to stimulate a calcium-plus-calmodulin-dependent cAMP phosphodiesterase from bovine brain; (b) the inhibitory effect of trifluoperazine in this process; (c) the stimulation of the [Ca2+ + Mg2+)-ATPase from human erythrocyte membranes. 4. Maximum effect of modulator protein in the above mentioned reactions was achieved at micromolar calcium concentrations. PMID- 6279358 TI - Disulfiram treatment of three patients with nickel dermatitis. AB - Diethyldithiocarbamate chelates nickel and accelerates its excretion in the urine. Disulfiram (Antabuse) splits into two molecules of sodium diethyldithiocarbamate after absorption. Oral administration of disulfiram tablets starting with 50 mg a day with a gradual increase in dosage to 100 mg more than than twice a day readily cleared the eczema in three patients with severe and long-standing nickel contact dermatitis. PMID- 6279359 TI - Dog response to inactivated canine parvovirus and feline panleukopenia virus vaccines. AB - Inactivated canine parvovirus (CPV) and inactivated feline panleukopenia virus (FPV) vaccines were evaluated in dogs. Maximal serologic response occurred within 1-2 weeks after vaccination. Antibody titers then declined rapidly to low levels that persisted at least 20 weeks. Immunity to CPV, defined as complete resistance to infection, was correlated with serum antibody titer and did not persist longer than 6 weeks after vaccination with inactivated virus. However, protection against generalized infection was demonstrated 20 weeks after vaccination. In unvaccinated dogs, viremia and generalized infection occurred after oronasal challenge with virulent CPV. In contrast, viral replication was restricted to the intestinal tract and gut-associated lymphoid tissue of vaccinated dogs. Canine parvovirus was inactivated by formalin, beta-propiolactone (BPL), and binary ethylenimine (BEI) in serum-free media; inactivation kinetics were determined. Formalin resulted in a greater loss of viral HA than either BEI of BPL, and antigenicity was correspondingly reduced. PMID- 6279360 TI - The spectrum of organoid nevi. AB - Two patients in whom tumors developed in organoid nevi are reported. The first patient, a 50-year-old man, had a trichilemmoma arising from an organoid nevus on the scalp. The second patient, a 68-year-old woman, had a basal cell epithelioma, sebaceous epithelioma, syringocystadenoma papilliferum, and a trichilemmoma arising from an organoid nevus on the face. PMID- 6279361 TI - Lung cancer among Navajo uranium miners. AB - Lung cancer has been a rare disease among the Indians of the southwestern United States. However, the advent of uranium mining in the area has been associated with an increased incidence of lung cancer among Navajo uranium miners. This study centers on Navajo men with lung cancer who were admitted to the hospital from February 1965 to May 1979. Of a total of 17 patients with lung cancer, 16 were uranium miners, and one was a nonminer. The mean value of cumulative radon exposure for this group was 1139.5 working level months (WLMs). The predominant cancer type was the small cell undifferentiated category (62.5 percent). The low frequency of cigarette smoking in this group supports the view that radiation is the primary cause of lung cancer among uranium miners and that cigarette smoking acts as a promoting agent. PMID- 6279362 TI - Japanese B encephalitis virus infection of horses during the first epidemic season following entry into an infected area. PMID- 6279363 TI - Epidermal growth factor: a critical factor in fetal maturation? AB - Epidermal growth factor (EGF) infused over 3-14 days into fetal sheep of 110-125 days gestation resulted in a number of morphological and endocrine changes. Striking hypertrophy of the skin, wool follicles and their accessory structures was seen, together with a reduction in the ratio of secondary to primary follicles and degenerative changes in wool fibres associated with shedding of fibres. Adrenal, thyroid, liver and kidney weights were increased while thymus weight was decreased. The increase in adrenal size resulted from cortical hypertrophy and was associated with increased cortisol secretion. Thyroid hypertrophy was accompanied by an increase in colloid stores, decreased plasma thyroxine and reverse triiodothyronine (T3) concentrations, unchanged plasma T3 and thyroid-binding globulin and raised thyrotropin (TSH) levels. Thyrotropin receptor affinity and content per gram of tissue were unchanged. Fetal and maternal plasma prolactin and growth hormone levels, and fetal plasma placental lactogen levels, were unchanged, although there was a significant rise in maternal plasma placental lactogen concentrations with high doses of EGF. Other maturational parameters such as switching from fetal to adult haemoglobin and liver glycogen content were unaffected. PMID- 6279364 TI - Development of peptidergic systems in the rat brain. AB - The brain contains a large variety and number of peptides some of which were known earlier as hypothalamic hormones (vasopressin, oxytocin, luteinizing hormone-releasing hormone, thyrotropin-releasing hormone, somatostatin) or as pituitary hormones (the family of opiomelanocortins), while others, not primarily known as hypothalamic or pituitary hormones, may also have endocrine effects (substance P, angiotensin II, neurotensin, bombesin, vasoactive intestinal peptide (VIP), gastrin-cholecystokinin, glucagon, carnosine, bradykinin). These peptides, which form a new class of putative neurotransmitters, are present early in brain development and show important sex differences in both their pattern of innervation and their effects. Their peripheral effects may include intrauterine growth of the placenta and fetus, the timing of birth, acceleration of the course of labour and responses to haemorrhage (redistribution of cardiac output and stimulation of blood cell formation). Endogenous peptides are probably involved in brain development, which may explain their general, permanent and sex dependent effects when given in the period of rapid brain development. Although peptides might in the future be useful for stimulating recovery from retarded brain development, at present one should be aware of the potential dangers of their use in, for example, obstetrics. PMID- 6279365 TI - Development of the fetal lung. AB - The development of the fetal lung compared to that of other organs is unusual in the degree of its dependence on extrinsic stimuli. When the space available to the growing lung is limited by space-occupying lesions or when the diaphragm is paralysed, lung growth is markedly impaired. The relationship of lung volume to growth may depend on lung distension. Lung hypoplasia associated with experimental procedures causing inhibition or blunting of fetal breathing movements suggests that the distending forces may be generated by these movements. Maturation is less dependent on distension and more dependent on the hormonal environment. Distensibility and stability of the lung in fetal sheep develops rapidly within a few days of birth and correlates strongly with the plasma cortisol concentration. Hypophysectomy retards mutation which is restored by infusing adrenocorticotropin but not cortisol into the fetus. The hormones mainly responsible for controlling the various aspects of maturation probably include cortisol, iodothyronines and catecholamines but the interrelationships of these hormones and the extent of involvement of other hormones is uncertain. PMID- 6279366 TI - The development of fetal adrenal function. AB - The response profiles of fetal sheep adrenals to tropic stimulation have been examined ih vivo and in vitro. Isolated adrenal cells from sheep fetuses in early pregnancy (Day 50) reduced cortisol in response to ACTH, dibutyryl cyclic AMP and GTP. The response was minimal on Day 100, but reappeared near term. 17 alpha Hydroxyprogesterone was converted to cortisol by adrenals of all ages, but pregnenolone and progesterone were converted to cortisol only in early and late, but not mid-pregnancy. These studies suggested that the mid-gestation loss of fetal adrenal responsiveness was associated with post-receptor/adenylate cyclase events and involved loss of 17 alpha-hydroxylase activity. Fetal adrenal function was activated by exogenous ACTH in vivo, and was reflected in an increase in the ratio of cortisol to corticosterone in fetal plasma and in augmented cortisol output in vitro from dispersed fetal adrenal cells. The results were consistent with an effect of ACTH administration on 17 alpha-hydroxylation. Fetal pituitary cells, prostaglandin E2, alpha-MSH and term placental extract are other potential (sources of) corticotropins, although further studies are required to delineate the nature and origin of the active substances, and/or their primary sites of action. PMID- 6279367 TI - Maturation of hypothalamic-pituitary function in the ovine fetus and neonate. PMID- 6279368 TI - Regulation of hormone production in the human feto-placental unit. AB - Hormone production in the human feto-placental unit has been studied extensively yet relatively little is known about the regulatory mechanisms involved. A tissue culture approach has been used to examine the effect of potential controlling factors on steroid production by the human mid-term fetal adrenal and mid-term and term placenta. Adrenal. The pituitary peptides corticotropin (ACTH) and alpha melanocyte-stimulating hormone (alpha-MSH) had the most significant influence on adrenal steroidogenesis in both the fetal and definitive zones. Their effects were not identical: they enhanced dehydroepiandrosterone sulphate (DHA-S) production in a comparable manner but alpha-MSH had much less of a stimulatory effect on cortisol biosynthesis. Medium from homologous fetal pituitary cultures mimicked the effects of alpha-MSH rather than ACTH. Homologous placental culture medium and progesterone enhanced only cortisol production and only in the fetal zone cells. These results demonstrate that specific fetal pituitary and placental factors influence fetal adrenal activity and suggest a functional zonation of the fetal adrenal. Placenta. DHA, DHA-S and 16-hydroxy-DHA stimulated oestrogen biosynthesis while high concentrations of DHA and DHA-S (but not 16-hydroxy-DHA) inhibited progesterone production. Luteinizing hormone-releasing hormone (LRH) inhibited both oestrogen and progesterone biosynthesis. Placental steroidogenesis can therefore be influenced not only by the fetus, through its increasing adrenal output of oestrogen precursors, but also by factors originating within the placenta itself. PMID- 6279369 TI - Colorectal linitis plastica. AB - Primary linitis plastica of the colon is an uncommon tumor, with only 22 cases having been previously reported. Experience with two men and one woman between 17 and 55 years of age, portraying a spectrum of clinical presentations ranging from rectal bleeding to large-bowel obstruction, is reviewed. Clinical and radiologic characteristics and histologic features helpful in making the diagnosis are detailed. Though the prognosis is grim, resective surgery, including oophorectomy in women, should be undertaken. Adjuvant therapy has been disappointing. PMID- 6279370 TI - Maturation-related changes in catecholamine-dependent cyclic AMP and protein kinase in the rabbit myocardium. AB - It has been shown that the sensitivity to isoproterenol of myocardial inotropic response increases with growth in rabbits. In the present study we attempted to delineate the biochemical mechanisms involved in this maturation-related change. We investigated cyclic AMP generation and protein kinase activation by isoproterenol in the ventricular myocardium of rabbits aged 1 day, 1 week, and 1 month. In contrast to an increase in inotropic response, both the sensitivity to isoproterenol of cyclic AMP generation and the sensitivity to exogenous cyclic AMP of protein kinase activation decreased with maturation from 1 day to 1 month of age. ED50 of isoproterenol (mean +/- SE) on cyclic AMP generation in myocardial slices was 1.83 +/- 0.42 x 10(-7) M for 1-day-old, 3.70 +/- 0.61 x 10( 7) M for 1-week-old, and 8.32 +/- 1.20 x 10(-7) M for 1-month-old rabbits. Likewise, the ED50 of isoproterenol on adenylate cyclase activation was 0.65 +/- 0.11 x 10(-7) M, 2.04 +/- 0.32 x 10(-7), and 15.2 +/- 2.5 x 10(-7) M in 1-day, 1 week- and 1-month-old rabbits, respectively. The ED50 of cyclic AMP on protein kinase activation was 1.50 +/- 0.64 x 10(-8) M for 1-day-old and 7.08 +/- 1.52 x 10(-8) M for 1-month-old rabbits. This apparent discrepancy between inotropic response and biochemical response in the sensitivity to isoproterenol indicates that the biochemical mechanism of the maturation-related change in inotropic response was at a step or steps distal to protein kinase activation. PMID- 6279371 TI - Ontogeny of myocardial alpha 1-adrenergic receptor in the rat. AB - The alpha 1-adrenergic specific radioligand [3H]-prazosin and the beta-adrenergic radioligand (-)-[3H]-dihydroalprenolol [(-)-[3H]-DHA] were used to describe the postnatal ontogeny of alpha 1- and beta-adrenergic receptors in particulate fractions of rat ventricular myocardium. The number of [3H]-prazosin binding sites increased 4- to 5-fold during the first 15 days of age from 35 +/- 9 to 163 +/- 5 fmol . mg-1 protein (p less than 0.01), and thereafter decreased progressively with age, reaching 83 +/- 5 fmol . mg-1 protein in the adult myocardium. (-)-[3H]-DHA binding to the same tissue decreased progressively at 1 day of age from 61 +/- 6 in the newborn to 37 +/- 4 fmol . mg-1 protein in the adult (p less than 0.01). Affinities (KD) for [3H]-prazosin or (-)-[3H-DHA were approximately 0.13 and 1.8 nM, respectively, and did not change with advancing age. We conclude that the postnatal development of the alpha 1-adrenergic receptors is distinct from that of the beta-adrenergic receptor in the rat ventricular myocardium. Possible explanations for this difference are discussed in relation to cardiac sympathetic innervation and cardiac growth. PMID- 6279372 TI - Patterns of corticotropic cells in the adult human pituitary in Cushing's disease. AB - The overall anatomical distribution of specifically stained corticotropic cells has been studied in pituitaries obtained at autopsy from 9 patients with Cushing's disease. Three patterns have been demonstrated consistent with the theory that it is not a single entity, but that some cases are of primary pituitary aetiology, whereas others are the result of hypothalamic or central nervous system dysfunction. The junctional corticotropic cells appear to react differently from those in the anterior pituitary to some stimuli, supporting the hypothesis that these cells may represent the remnants of the pars intermedia in the human adult, and suggesting that they may have some as yet unidentified physiological function. PMID- 6279373 TI - Recent progress in the histological typing of human breast tumours. AB - Recently, new concepts in the histological typing of human mammary carcinomas have been proposed. The basic subdivision into the two major categories, i.e. ductal and lobular carcinomas, remains valid. In so far as lobular carcinomas are concerned, histological variants have been recognized: invasive carcinoma without an in situ component as well as solid and signet-ring cell forms. Special types of duct carcinoma have been defined with more accuracy, those deserving special attention being, scirrhous, tubular and secretory forms. The validity of the diagnosis of so-called apocrine carcinoma is discussed, as is the place of mucus secreting tumours in the histologic classification. PMID- 6279375 TI - [Segmental resection of thoracic trachea for cylindroma--a case report (author's transl)]. PMID- 6279374 TI - Alveolar trapping in pulmonary carcinomas. AB - Sixty surgically resected pulmonary carcinomas have been studied by light and electron microscopy for engulfed alveolar cavities within the tumour mass. Alveoli had been incorporated in twelve cases, ten epidermoid (squamous) carcinomas and two adenocarcinomas. They were lined by granular pneumocytes and cells transitional between granular and membranous pneumocytes. The lining cells showed a positive reaction to alkaline phosphatase. Cellular debris, leucocytes and mucus could be seen within the residual lumen. It is concluded that alveoli commonly found within pulmonary carcinomas. Accurate histological classification requires these to be differentiated from true neoplastic spaces. PMID- 6279376 TI - [Benign mixed mesodermal tumor of the broad ligament (author's transl)]. PMID- 6279377 TI - [Sensitivity of the retinal pigment epithelium of newborn rats to the melanotropic action of adrenocorticotropic hormone]. PMID- 6279378 TI - [New site-specific endonuclease from Streptomyces: SGRII]. PMID- 6279380 TI - [Foodborne intoxications by Clostridium perfringens type A in chicken salad (author's transl)]. PMID- 6279379 TI - Clonidine and glucose intolerance. AB - Changes in carbohydrate metabolism were observed in a diabetic, hypertensive patient managed with clonidine. After a slight increase in the clonidine dose, his blood sugar control deteriorated. However, when the clonidine was withdrawn, the glucose intolerance subsided. Because clonidine preferentially binds alpha 2 subtype receptors, we investigated animal pancreatic tissue by radioligand binding technique and found it to contain alpha-adrenergic receptors predominantly of the alpha 2-subtype. In consideration of the response from withdrawal of clonidine and the results of our radioligand studies, wer concluded the glucose intolerance seen in this patient was most likely due to the specific action of clonidine on alpha 2-pancreatic receptors. PMID- 6279381 TI - [Corona-, rota- and parvovirus infection in calves from a clinical point of view (author's transl)]. PMID- 6279382 TI - Failure of ethanol or isopropanol pretreatment to affect carbon tetrachloride induced inhibition of hepatic microsomal calcium pump activity. AB - The calcium pump of liver microsomes is sensitive to CCl4 metabolism in vitro and in vivo. Treatment of rats with alcohol might involve an enhancement of the action of low doses of CCl4 on rat liver microsomal calcium pump. It was found that treatment of fasted rats with isopropanol or ethanol potentiated the increase of serum glutamic oxaloacetic transaminase elicited by CCl4. Alcohol pretreatment alone had no effect on calcium pump activity. After CCl4 alone, the calcium pump was 50% inhibited at 20 min, and 85% inhibited at 1 hr. Pretreatment with either alcohol had no effect on either rate of decline or extent of CCl4 dependent depression of the liver microsomal calcium pump. The mechanism of alcohol potentiation of CCl4-induced hepatotoxicity probably resides in alteration of processes developing after the initial events of CCl4 metabolism. PMID- 6279383 TI - [Trial of a new drug in Raynaud's disease. Clinical and thermographic control]. PMID- 6279384 TI - Volume conduction of motor unit potentials from different human muscles to long distances. PMID- 6279385 TI - Facial reflexes of short latency. PMID- 6279386 TI - Effect of MgATP on cAMP-dissociation kinetics of lactating rat mammary gland. AB - Mammary gland cytosols exhibit temperature-dependent interconversion of cAMP dissociation rates from low to high affinity (k-1 = 0.14 min-1 at 0 degree C to k 1 = 0.02 min-1 at 24 degrees association). This interconversion corresponds to a change from a site 2 to a site 1 cAMP-dissociation rate for the type II cAMP dependent protein kinase in mammary gland cytosols. This report presents data which indicates a requirement for MgATP in the temperature-dependent interconversion of cAMP-dissociation rates. The effect of MgATP on the generation of the high affinity state was observed at 24 degrees C but not 0 degree C association. The effect of MgATP was not mimicked by equimolar MgAMP-PNP, but did require an intact type II protein kinase holoenzyme which can undergo autophosphorylation of its regulatory subunit. The effect of MgATP was reproduced with partially purified preparations of beef heart type II protein kinase. These results suggest that MgATP may act through autophosphorylation of the type II holoenzyme. The data suggest a novel role of MgATP in the regulation of cAMP binding to cAMP-dependent protein kinase II. PMID- 6279387 TI - Comparative effects of fibrous and nonfibrous minerals on cells and liposomes. PMID- 6279388 TI - The effects of sample preparation and measuring techniques on the shape and shape characterization of mineral particles: the case of wollastonite. PMID- 6279389 TI - Alterations in hepatic 5'-nucleotidase in the tumor-bearing rat. AB - The effect of the growth of the Walker 256 carcinoma on the level of 5' nucleotidase and alkaline phosphatase in the whole liver and in an isolated hepatocyte membrane preparation of its host was investigated. Alkaline phosphatase activities of whole liver and plasma membrane were increased approximately 5-fold by tumor growth. A 50% decrease in whole liver 5' nucleotidase activity was observed in tumor-bearing rats while the 5' nucleotidase activity per milligram membrane protein was unaltered. Tumor growth would therefore appear to affect a pool of 5'-nucleotidase which is not associated with the plasma membrane. PMID- 6279390 TI - Alterations in the activities of intestinal enzymes in vitamin-C-deficient guinea pigs. AB - The effect of vitamin C deficiency on various enzymes of the intestinal epithelium has been studied in guinea pigs. Brush border sucrase and alkaline phosphatase activities were considerably enhanced (p less than 0.001), but leucine aminopeptidase levels were reduced in scorbutic animals compared to the control group. There was essentially no change in the activity of maltase under these conditions. Kinetic studies with sucrase and alkaline phosphatase in control and scorbutic animals revealed that augmentation of the enzyme activities in scurvy is due to enhanced enzyme contents. Lactate dehydrogenase, succinate dehydrogenase, glucose-6-phosphatase and Mg+2 ATPase also exhibited reduced activities in the intestine of vitamin-C-deficient animals. Observed alterations in the activities of intestinal enzymes in scurvy were restored to control levels upon feeding of vitamin C to scorbutic guinea pigs. PMID- 6279391 TI - Effects of nutritional deficiencies during neonatal and post-weaning period on rat intestinal phytase and phosphatase activities. AB - Studies were made of the effects of pre- and post-weaning undernutrition and/or protein deficiency on intestinal phytase and phosphatase activities in albino rats and reversibility of the same by subsequent dietary rehabilitation. Neonatal undernutrition induced by rearing the pups in litters of 16 caused a marked decrease in alkaline phytase activity (as compared to those reared in litters of 8), while acid phytase activity decreased to a lesser extent and acid and alkaline phosphatase activities did not change. When neonatally undernourished rats were subsequently continued on a 4 or a 20% protein diet in restricted amounts (2.5 g/day) for 6 weeks the decreases in the alkaline phytase activity but not in that of acid phytase were further aggravated. Acid and alkaline phosphatases were not influenced by these treatments either. On dietary rehabilitation of these rats for subsequent 6 weeks on a 20% protein diet (ad libitum) acid and alkaline phytase activities of intestine recovered partially. These studies indicate the importance of alkaline phytase activity as a marker of intestinal maturation and is also suggestive of interrelationships between nutrition, intestinal development and its alkaline phytase activity. PMID- 6279392 TI - Glycolytic activity of rat aorta after exercise. AB - Activities of glycolytic enzymes in the aorta were investigated in female Wistar rats. There were two groups of rats; one served as the control (sedentary rats), while the other group was forced to run on a treadmill for 10 weeks. In the control animals, the activities of hexokinase, phosphofructokinase and aldolase were relatively lower than those of the other glycolytic enzymes (phosphoglucose isomerase, lactate dehydrogenase and pyruvate kinase). After exercise, the activity of phosphofructokinase increased by 15%, whereas the other enzymatic activities were much the same as in the controls. Within the limits of the experiments, the increased percentage of phosphofructokinase was statistically significant (p less than 0.05). Since phosphofructokinase is a putative rate limiting enzyme, this enzymatic activation may indicate that glycolytic activity in the rat aorta is enhanced during and after running exercise. PMID- 6279393 TI - Two-dimensional NMR spectroscopy of siomycin A. Proton--carbon-13 chemical shift correlation. AB - A trial application of a recent two-dimensional nuclear magnetic resonance experiment to the polypeptide antibiotic siomycin A is described. Proton--carbon 13 chemical shift correlation measures the proton and carbon-13 chemical shift for each directly bonded CH group in a molecule, in a single experiment. The resultant map of correlated chemical shifts enables the carbon-13 spectrum to be assigned directly from the known proton shifts, and allows individual proton signals to be identified without problems of overlap. The signal-to-noise ratio available from such techniques should enable their application to aqueous protein solutions using currently available high-field spectrometers. PMID- 6279394 TI - Light-induced interactions between rhodopsin and the GTP-binding protein. Relation with phosphodiesterase activation. AB - The existence of rapid light-induced changes of light scattering in suspensions of bovine rod outer segment membranes has been described previously [H. Kuhn et al. (1981) Proc. Natl Acad. Sci. USA, 78, 6873-6877]. The signal observed in the presence of GTP has been interpreted as being related to the rhodopsin-catalyzed exchange of GTP for GDP bound to the GTP-binding protein, i.e. to the formation of the activator of the cGMP phosphodiesterase [B.K.K. Fung et al. (1981) Proc. Natl Acad. Sci. USA, 78, 152-156]. We have tested this interpretation in the present paper by investigating the relation between the light-scattering signal and the activity of the phosphodiesterase using rapid recording techniques for both processes. All the results obtained are consistent with the above hypothesis. The amplitude of the light-scattering signal and the activity of the phosphodiesterase are shown to present the same dependence upon the flash intensity and upon the concentration of GTP or its analog guanosine 5'-[beta, gamma--imido]triphosphate (p[NH]ppG). The results suggest that the GTP-binding protein possesses one high-affinity p[NH]ppG-binding site (Kd much less than 0.1 microM). At high concentrations of GTP or p[NH]ppG the phosphodiesterase is activated in the dark and the light-scattering signal is correspondingly reduced; both effects are prevented by previous incubation with guanosine 5'-[beta thio]diphosphate (p[S]pG). PMID- 6279395 TI - Temperature dependence of the activity of DNA-modifying enzymes: endonucleases and DNA ligase. AB - The activities of 17 endonucleases: the restriction endonucleases AvaI, Bam HI, EcoRI, HindIII, PstI and SalI, which cleave pBR322 DNA once: AluI, AvaII, CfoI, HaeIII, HhaI, HinfI, HpaII and TaqI, which cut pBR322 DNA several times, and three 'unspecific' nucleases (S1 nuclease, staphylococcal nuclease and DNase I from bovine pancreas) were determined between 0 degrees and 65 degrees C. The reaction was followed by the disappearance of covalently closed circular pBR322 DNA, using the alkaline ethidium fluorescence assay of Morgan et al. [Nucleic Acids Res. (1979) 7, 547-594]; the activity of T4 DNA ligase was similarly measured by the conversion of nicked circular DNA to closed circular DNA. For each enzyme, small aliquots of the same solution were incubated at different temperatures simultaneously in a temperature gradient device, resulting in a high relative precision. The experimental results are summarized by the simplest possible theoretical description, using linear or exponential kinetics and apparent activation energies Ea for the enzymatic reaction, Ei for the enzyme inactivation and Ti for the inactivation temperature. To a good approximation these three parameters suffice for describing the temperature dependence of the activity of most of the enzymes. PMID- 6279396 TI - Cytosolic and nuclear binding of 2,3,7,8-tetrachlorodibenzo-p-dioxin to the Ah receptor in extra-hepatic tissues of rats and mice. PMID- 6279397 TI - Metal coordination centres of class II cytochromes c. AB - The class II cytochromes Rhodospirillum molischianum cytochrome c', Rhodopseudomonas palustris cytochrome C556 and Agrobacterium tumefaciens (B2a) cytochrome c556 have been investigated with a variety of spectroscopic techniques. The cytochrome c' was found to be high-spin and the two cytochromes c556 were found to be mainly low-spin and sx-coordinate with the fifth and sixth ligands being histidine and methionine. The implications of the different types of iron coordination are discussed. PMID- 6279398 TI - Individual assignments of the amide proton resonances involved in the triple stranded antiparallel pleated beta-sheet structure of a long neurotoxin, Laticauda semifasciata III from Laticauda semifasciata. AB - THe characteristic feature of the crystal structure of erabutoxin b, a short neurotoxin from Laticauda semifasciata, and alpha-cobratoxin, a long neurotoxin from Naja naja siamensis, is the presence of a triple-stranded antiparallel pleated beta-sheet structure formed by the central and the third peptide loops. In the present study, we have studied the assignment of slowly exchangeable amide protons of Laticauda semifasciata III from L. semifasciata, using nuclear Overhauser effects (NOE) and spin-decoupling methods. The results show that nearly all of the slowly exchangeable amide protons are to be assigned to the back-bone amide protons, involved in the triple-stranded antiparallel pleated beta-sheet structure, indicating that this sheet is stable in 2H2O solution. In contrast, the amide protons in short neurotoxins are readily exchangeable under the same experimental condition, suggesting that long neurotoxins have a more rigid sheet structure than short ones. This rigidity may come from the hydrophobic and hydrogen bond interaction between the central loop and the tail, which is not present in short neurotoxins. Since the functionally important residues are located on this beta-sheet, the different kinetic properties of the neurotoxins are well correlated with the difference in the rigidity of the beta sheet. PMID- 6279399 TI - Radiology of uncommon esophageal neoplasms. AB - The radiographic appearance of 16 uncommon primary esophageal tumors (4 adenoid cystic carcinomas, 3 adenoacanthomas, 4 small cell carcinomas, 2 adenopapillary carcinomas, adenocarcinoma, mucoepidermoid carcinoma and malignant melanoma), is presented. These tumors were found in a retrospective study of 270 esophageal neoplasms. Sufficient radiographic, histologic and clinical data was available to substantiate primary esophageal origin of the tumors. Cylindromas tender to have a smooth surface and either funneled or polypoid appearance. None of the tumors, however, had radiographic appearance readily distinguishable from variable patterns of the common esophageal carcinomas. The age and sex distribution, location as well as prognosis of the rate adenocarcinomas appear to be similar to squamous cell carcinomas. The small cell carcinomas of the esophagus only had an average prognosis of 4 months. The x-ray examination was of little value in predicting local tumor spread, mediastinal metastases or prognosis. PMID- 6279400 TI - Efficiency of different radiodiagnostic techniques in pancreatic disorders. AB - Using figures from the literature and from our own case material, the diagnostic efficiency of the barium meal, computerized tomography, angiography, ERPC and biopsy have been compared in acute and chronic pancreatitis, pancreatic and papillary carcinoma and islet cell tumour. In acute pancreatitis, CT is the method of choice. In chronic pancreatitis, ERPC has the highest diagnostic accuracy. Furthermore, it is capable of giving an indication for operative intervention, and facilitating the choice of operative method. In pancreatic carcinoma, combination of ERPC, angiography and/or biopsy provides the highest diagnostic accuracy. Angiography and ERPC can provide early diagnosis, and angiography and CT can predict the operability of a tumour. CT is also the most important of the non-invasive diagnostic techniques. Carcinoma of the papilla is easily and reliably recognized by endoscopy. For islet cell tumours, angiography still remains the method of choice. PMID- 6279401 TI - New aspects of the uptake of thallous ion into myocardial tissue. PMID- 6279402 TI - Alkaline phosphodiesterase I of cultured mouse peritoneal macrophages. Distribution between cell surface and cytoplasm, turnover rate, and effects of colchicine. AB - Mouse peritoneal macrophages were cultivated in vitro and analyzed for activity of alkaline phosphodiesterase I. The specific enzyme activity was 2 to 3 times higher in thioglycollate-elicited than in resident cells and raised slowly for at least 3 days of culture in both cell types. Following phagocytosis of polystyrene latex beads the enzyme activity remained unchanged. About 90% of the activity was lost by treatment of the cells with the diazonium salt of sulfanilic acid (DASA) for 30 min at 37 degrees C. At 4 degrees C the corresponding degree of inactivation was only 40 to 50%. This temperature-dependent variation in the accessibility of the enzyme to inactivation by DASA could be related to the high rate of endocytosis in macrophages. During a 30 min incubation with DASA at 37 degrees C, large amounts of surface membrane will be internalized and replaced from the interior of the cell. Surface membrane is thus transferred into intracellular membrane and vice versa. Moreover, these membranes will be exposed to DASA independent of their location at the start of incubation. At 4 degrees C no comparable membrane traffic will take place. Treatment of the cells with cycloheximide at concentrations that inhibited protein synthesis by 90% or more left the enzyme activity essentially unaltered, indicating a slow turnover rate. On the basis of these findings, it is suggested that about half of the alkaline phosphodiesterase I activity of cultured macrophages is located in the plasma membrane. The other half is believed to be present in endocytic vesicles, lysosomes, and other as yet unidentified organelles that participate in the circulation of membrane constituents between the plasma membrane and the interior of the cell. PMID- 6279403 TI - Mechanism of function of the plasma membrane receptor specific for galactose terminated glycoproteins in hepatocytes and after transfer to mouse L-cells. PMID- 6279405 TI - Pulmonary insufficiency in the rat after intravascular coagulation and inhibition of fibrinolysis. I. Studies on some pathogenetic mechanisms. PMID- 6279404 TI - 67Gallium scintigraphy in myocarditis. PMID- 6279406 TI - Phase II study of oral VP-16-213 in hepatocellular carcinoma. PMID- 6279407 TI - Immunoperoxidase staining for carcinoembryonic antigen in small cell carcinoma of the lung. PMID- 6279408 TI - Estrogen receptors in cystosarcoma phyllodes of the breast. PMID- 6279409 TI - Long-term survival five years or more after combination chemotherapy and radiotherapy for small cell lung carcinoma. PMID- 6279410 TI - Calcium channel blockers: a pathophysiologically based antihypertensive treatment concept for the future? PMID- 6279411 TI - Beneficial action of a new opiate antagonist (Win 44,441-3) in hemorrhagic shock. AB - The new opiate antagonist Win 44,441-3 (-)-isomer was infused intravenously in cats at a rate of 2 mg . kg-1 . h-1 to determine its effect in hemorrhagic shock. Hemorrhaged cats treated with Win 44,441-3 maintained post reinfusion mean arterial blood pressure (MABP) at a higher value compared to cats receiving only the vehicle. Final MABP was 70 +/- 11 mm Hg for cats receiving vehicle compared to 103 +/- 7 mm Hg for cats receiving Win 44,441-3. These values represent 60 +/- 9% and 85 +/- 6% of initial MABP for the vehicle- and Win 44,441-3-treated cats respectively. Win 44,441-2 (+)-isomer, the inactive stereoisomer of Win 44,441-3, was also infused at 2 mg . kg-1 . h-1 in cats subjected to hemorrhagic shock. The final pressure in this group was 72 +/- 8 mm Hg which is 61 +/- 8% of the initial pressure for this group. Win 44,441-3 and Win 44,441-2 were both ineffective in moderating increases in circulating lysosomal hydrolase activity in shocked cats. Neither isomer stabilized lysosomal membranes or retarded proteolysis in vitro. Plasma myocardial depressant factor was significantly reduced by the opiate antagonist, Win 44,441-3 during shock. Our results show that the systemic infusion of an opiate antagonist improves the hemodynamic state of cats subjected to hemorrhagic shock while the (+)-isomer which lacks opiate antagonist activity produces no such improvement. PMID- 6279412 TI - Effects of tetrazole derivatives on [3H]diazepam binding in vitro: correlation with convulsant potency. AB - A series of 8 tetrazole derivatives which differ more than one hundred-fold in their potencies as convulsants were tested for their abilities to inhibit [3H]diazepam binding to benzodiazepine receptors in vitro. The concentrations of drug necessary to inhibit 50% of specifically bound [3H]diazepam ranges from 18 micro M for undecamethylenetetrazole to 20 mM for trimethylenetetrazole. A comparison of the minimum convulsive doses for the 8 tetrazole derivatives tested with their relative potencies in displacing [3H]diazepam binding in vitro revealed a highly significant correlation (r = 0.98, P less than 0.001). In contrast, several representative tetrazole derivatives were found to have no inhibitory effects on beta-adrenergic, alpha-adrenergic or muscarinic cholinergic receptors in the same membrane preparation. These results suggest that pentamethylenetetrazole and related tetrazole derivatives may elicit their convulsant effects by interaction with the benzodiazepine receptor. PMID- 6279413 TI - Ketamine inhibition of ligand binding to cholinergic receptors and ion channels. AB - Ketamine inhibited the binding of [3H]perhydrohistrionicotoxin and [3H]phencyclidine to the ion channel associated with Torpedo acetylcholine receptors with ID50 values between 11 and 84 micro M, but had not effect on [3H]acetylcholine or [3H]d-tubocurarine binding to the receptor itself. Ketamine's affinity for the ion channel was increased 3-5 fold by receptor agonists. Ketamine also inhibited muscarinic cholinergic receptors with ID50 between 28 and 38 micro M. These results are consistent with biophysical evidence that ketamine interferes with neuromuscular transmission through blockade of the ion channel. PMID- 6279414 TI - [3H]Tyr-D-Ser-Gly-Phe-Leu-Thr: a specific probe for the delta-opiate receptor subtype in brain membranes. PMID- 6279416 TI - Contact relationships between chick embryo cells growing in monolayer culture after infection with Rous sarcoma virus. PMID- 6279415 TI - Biochemical and morphological characterization of glycogen-storing epithelial liver cell lines. PMID- 6279417 TI - Isolation of parenchymal cell-derived particles from non-parenchymal rat liver cell preparations. PMID- 6279418 TI - Prolactin stimulation of ornithine decarboxylase activity in cultured rat mammary tumors. PMID- 6279419 TI - Distribution of a 100K dalton glucose-regulated cell surface protein in mammalian cell cultures and sectioned tissues. PMID- 6279420 TI - Serum protein electrophoretic pattern and angiotensin-converting enzyme in sarcoidosis. AB - S-angiotensin-converting enzyme (SACE), s-protein electrophoretic pattern and ESR were examined in 61 patients with sarcoidosis. Among 46 patients with active disease there was a significant elevation of ESR, alpha 2-globulin, and gamma globulin, but the deviations from healthy controls were generally small and the pattern was unspecific, without correlation to clinical or roentgenographic features. Elevation of SACE was confined to active sarcoidosis, in which group elevation was noted in 59% of the patients. SACE was positively correlated to total protein and gamma-globulin. This correlation may not be causal but rather an expression of two different cellular activities in the disease, involving the macrophage-system and the B-lymphocytes, respectively. PMID- 6279421 TI - Activity of angiotensin-converting enzyme in pleural fluid and serum in non sarcoid, non-tuberculous pleural effusion. AB - The activity of angiotensin-converting enzyme (ACE) was measured in pleural fluid (PACE) and serum (SACE) from 25 patients with pleural effusion due to cancer or non-malignant diseases. In all patients SACE was within normal limits. One patient with a rheumatic pleuritis had a PACE/SACE ratio greater than 3.0, but among the remaining 24 patients PACE was 12.1 +/- 6.5 U/ml (mean +/- 1 SD), and SACE was 21.9 +/- 6.4 U/ml (P less than 0.005), with a PACE/SACE ratio of 0.58 +/ 0.31. SACE and PACE were positively correlated in patients with exudative pleuritis (P less than 0.01). In hydrothorax due to congestive heart failure a very low PACE/SACE ratio was found. Enzyme activity or ratio was correlated to protein content (P less than 0.02) but not to aetiology of pleural effusion, presence of malignant cells, leucocyte- pr erythrocyte content. In general, measureable CE activity can be detected in pleural fluid, but this finding seems at present to have no clinical significance in these conditions. PMID- 6279422 TI - Interactions between motor units in modulating discharge patterns of primary muscle spindle endings. AB - Discharge patterns of group Ia afferents of muscle spindles in the (isometric) cat semitendinosus muscle were recorded when subjected to the influence of contractions of three motor units elicited by stimulation of three alpha efferents with different regular or stochastic stimulus patterns. In order to study conditioning interactions between motor unit contractions, defined time constellations of stimuli to different motor units were isolated from the stimulus trains by electronic means. This technique was also applied to discharge patterns of two Ia fibres to isolate "correlated discharges." The effects of defined time constellations of stimuli upon discharge patterns of single Ia afferents or "correlated discharges" of pairs of Ia afferents were assessed by poststimulus time histograms (PSTHs). It turned out that the influence exerted by one motor unit on spindle discharges was more or less strongly modulated by actions of the other motor units, dependent on the relative timing of their contractions. These effects could in part be explained by the relative locations of motor units and spindles within the muscle. These results are discussed with respect to possible mechanisms of low-amplitude physiological tremor. PMID- 6279423 TI - Non-linear temporal summation by simple cells in cat striate cortex demonstrated by failure of superposition. AB - Simple cells in area 17 of the anaesthetized, paralysed cat were stimulated with stationary sinusoidal gratings whose contrast was temporally modulated in different ways. The response to a temporal waveform which was the sum of two sinusoids (1.25 Hz and 7.75 Hz) was compared with the response elicited by each component when presented alone. The responses to the high temporal frequency in the compound stimulus were relatively enhanced by the addition of the low temporal frequency; those to the low frequency were relatively depressed. PMID- 6279424 TI - Evidence against transmitter function of met-enkephalin and chemosynaptic impulse generation in "Merkel cell" mechanoreceptors. AB - Using the mechanoreceptors in the cat's sinus hair follicles as a model system the recent hypothesis (Hartschuh and Weihe 1980) was tested that Merkel cells are mechanoreceptive sensory cells releasing met-enkephalin as a neurotransmitter to initiate action potentials in the terminals of type I afferent fibres. Since the met-enkephalin antagonist naloxone, even in high doses, did not affect responses in any type I mechanosensitive afferent it is unlikely that a synaptic mechanism with the release of met-enkephalin underlies impulse generation in Merkel cell axon complexes, nor did naloxone influence the responses from other types of mechanoreceptors. PMID- 6279425 TI - The effect of gamma-aminobutyric acid on the potassium movements of rat cortical synaptosomes. AB - K+ fluxes of rat cortical synaptosomes were monitored by a K+ sensitive ion selective membrane electrode. The uptake phases of K+curves from control and experimental measurements were compared and statistically evaluated. GABA significantly reduced K+ uptake but it was able to reverse the decrease of K+ uptake caused by protoveratrine. In the absence of Ca2+ the K+ uptake was diminished; GABA had no effect on this inhibition. The effect of GABA on K+ take was partially reduced by bicuculline and abolished by diamino-butyric acid. The K+ content of synaptosomes decreased, while the NA+ content increased in the presence of GABA. The results are discussed in association with presynaptic depolarisation and with modulatory effects of GABA on the release of other transmitters. PMID- 6279426 TI - The spatial organization of the excitatory regions in the visual receptive fields of the pigeon's optic tectum. AB - The spatial location of the excitatory regions in the receptive field of cells in the pigeon's optic tectum was analyzed with light and dark edges moving at a constant velocity. The tectal cells were classified into two main groups: 1-cells showing spatially overlapping light and dark excitatory regions in their receptive field (60%); cells showing spatially separated light and dark excitatory regions in their receptive field (32%). A small number of cells discharged only to one sign of contrast. These results were confirmed by testing the cells with light bars of various widths. Latency studies were carried out with single edges moving at a series of constant velocities. In most cases, for any given cell the light and dark edge discharges were shown to have similar latencies. These results also indicate that the relative location of the excitatory regions in the receptive field of most tectal cells was not significantly affected by the latency of the discharges. PMID- 6279427 TI - Stimulus-induced changes in extracellular Na+ and Cl- concentration in relation to changes in the size of the extracellular space. AB - Extracellular Na+ - and Cl- -concentrations ([Na+]o, [Cl-]o) were recorded with ion-selective microelectrodes during repetitive stimulation and stimulus-induced self-sustained neuronal afterdischarges (SAD) in the sensorimotor cortex of cats. In all cortical layers [Na+]o initially decreased by 4-7 mM. In depths of more than 600 micrometer below the cortical surface such decreases usually turned into increases of 2-6 mM during the course of the SADs, whereas in superficial layers [Na+]o never rose above its resting level. [Cl-]o always showed an increase in the course of the SADs often preceded by an initial small decrease. The average increase at a depth of 1,000 micrometers was about 7 mM. [Cl-]o reached peak values at about the end of the ictal period, whereas [Na+]o reached its maximum shortly after the end of the SAD, at times when [K+]o was still elevated above the baseline concentration. These data indicate that the extracellular osmolarity can increase during SAD by up to 30 mM. Such an increase in osmolarity can be explained by an increase in the number of intracellular particles, caused by cleavage of larger molecules during enhanced metabolism. This could lead to cell swelling due to passive water influx from the extracellular space (ES). However, the resulting reduction of the size of the ES is calculated to be less than 10% for an increase in intracellular osmolarity by 30 mOsm. This value is too small as compared to previously measured ES-reductions under similar conditions (i.e., 30% reduction at 1,000 micrometers; Dietzel et al. 1980). Reductions of the size of the ES that accompany the observed changes in the ionic environment, are quantitatively explained on the basis of the extended glial buffering mechanism described in the preceding paper. PMID- 6279428 TI - Corticotrophin and corticosterone secretion following delta 1 Tetrahydrocannabinol, in intact and in hypothalamic deafferentated male rats. AB - Adult male rats, either intact (N) or bearing complete hypothalamic deafferentations (CHD), were injected with delta 1-tetrahydrocannabinol (THC: 5 mg/kg BW, IP). Forty-five minutes later, they were decapitated and trunk blood was collected for serum ACTH and corticosterone (CS) determinations. In the N animals, serum levels of both ACTH and CS were markedly elevated in the drug treated, as compared to the vehicle-treated group (approximately 8-fold and 10 fold, respectively). In CHD rats, on the contrary, THC administration did not significantly alter serum concentrations of either ACTH or CS. These results demonstrate (1) that acute treatment with THC stimulates the secretion of ACTH as well as of CS; and (2) that extrahypothalamic sites and/or neural pathways mediate this effect. PMID- 6279430 TI - Gastrin and the transforming protein of polyoma virus have evolved from a common ancestor. PMID- 6279429 TI - Foveal inhibition and facilitation caused by remote grating jerks: interaction between long-range and short-range effects. AB - Periodic oscillation of a luminance grating imaged upon the peripheral retina reduces the threshold visibility of a foveally presented test spot. This new effect has been named the "jerk effect". The present investigation is concerned with the effect of a single jerk of the remote grating on threshold sensitivity. Foveal sensitivity changes were measured for different delays between grating jerk and test spot presentation. For 0.38 degrees, 100 ms test spot, long-range transient inhibition was found for all delays, with a maximal effect between 0 and 30 ms delay. By combining the jerk effect with the Westheimer paradigm, both facilitatory and inhibitory long-range effects could be demonstrated. For facilitation to occur, it was necessary that the steady background extended into the sensitization zone of the Westheimer area. Inhibition was the only result for smaller backgrounds. Reduced visibility is consistent with the hypothesis that peripheral transient mechanisms inhibit foveal sustained mechanisms. Enhanced visibility indicates that thresholds depend on an interaction between foveal sustained and foveal-transient units. Transient peripheral stimulation and steady backgrounds of increasing diameter change the balance of inhibitory and facilitatory processes between these units. PMID- 6279431 TI - The effects of monoclonal antibodies against human growth hormone on hormone- receptor interactions. PMID- 6279432 TI - The topoisomerase activity of T4 amG39 mutant is restored in Mu lysogens. PMID- 6279433 TI - Pro-opiocortin converting activity in rat intermediate and neural lobe secretory granules. PMID- 6279434 TI - Discovery of a Ca2+- and calmodulin-dependent protein phosphatase: probable identity with calcineurin (CaM-BP80). PMID- 6279435 TI - Cyclic adenosine-5'-trimetaphosphate phosphorylates a histidine residue nearby the initiating substrate binding site of Escherichia coli DNA-dependent RNA polymerase. PMID- 6279436 TI - Structural and functional features of the interaction of cytochrome c with complex III and cytochrome c oxidase. PMID- 6279437 TI - Several nucleoside-3' and/or 5'-polyphosphates stimulate beta-galactosidase induction in Escherichia coli. PMID- 6279438 TI - alpha 2 HS glycoprotein binds to lymphocytes transformed by Epstein-Barr virus. PMID- 6279439 TI - Relative rates of 2- and 4-hydroxyestrogen synthesis are dependent on both substrate and tissue. PMID- 6279440 TI - The effect of benzo[a]pyrene on DNA synthesis and DNA polymerase activity of rat liver mitochondria. PMID- 6279441 TI - Inhibition of the purified and reconstituted calcium pump of erythrocytes by micro M levels of DIDS and NAP-taurine. PMID- 6279442 TI - Evidence against a requirement for phospholipid methylation in adenylate cyclase activation by hormones. Methyltransferase inhibitors do not impair cyclic AMP accumulation induced by glucagon or beta-adrenergic agents in rat hepatocytes. PMID- 6279443 TI - Heterogeneity of the catalytic activity arising from thyroidal cAMP-dependent protein kinase type II. PMID- 6279445 TI - Resonance Raman spectra of heme c and heme d1 in Pseudomonas cytochrome oxidase. PMID- 6279444 TI - Tissue-specific genes code for polypeptide VIa of bovine liver and heart cytochrome c oxidase. PMID- 6279446 TI - Structural features of the copper-depleted cytochrome oxidase from beef heart: iron EXAFS. PMID- 6279447 TI - Interaction between D-fructose-1,6-bisphosphate aldolase and triosephosphate isomerase. Mutual protection against perchloric acid denaturation. PMID- 6279448 TI - Phosphorylation of skeletal muscle myosin light chain kinase by the catalytic subunit of cAMP-dependent protein kinase. PMID- 6279449 TI - Effects of diphtheria toxin and other exotoxins on oxidant generation by human and murine phagocytes. AB - Bacterial exotoxins such as diphtheria toxin (D.T.), staphylococcal alpha toxin and Leucocidin can powerfully activate granulocytes and macrophages as detected by production of chemiluminescence in presence of Luminol. Production of superoxide by granulocytes and of prostaglandin E2 in macrophages is also stimulated by D.T. In contrast with the known resistance of rodent parenchymal cells to the diphtheria toxin, human and rodent leucocytes have similar sensitivities to D.T. PMID- 6279451 TI - Hydrolyzing activities on cAMP in E. coli B cells incubated in the presence of polyamines. AB - 1. Hydrolyzing activities on cAMP were investigated in E. coli B cells incubated in the presence or absence of 10(-5) M spermine, spermidine or putrescine. 2. Bacterial cells incubated in the presence of each of the investigated polyamines show an increase in the PDE (phosphodiesterase) activities already evident after 3 min of incubation, reaching the maximum between 5 and 10 min. disappearing between 15 and 25 min. 3. The stimulating effect is higher in the presence of lower (10(-6) M) substrate concentration (high affinity PDE activities). 4. Kinetic analyses carried out for the "high affinity" PDE activity, indicate that spermidine and putrescine are the most effective on increasing both the Vmax or the apparent Km. 5. Kinetic analysis carried out for the "low affinity" PDE activity, indicate that putrescine is the most active on increasing either the Vmax or the apparent Km and that spermine and spermidine have both quantitatively and qualitatively comparable effects. 6. Analyses, by TLC, of the products of the hydrolytic activities on cyclic AMP (5'-adenosine monophosphate (5'-AMP), adenosine, inosine) indicate that the incubation in the presence of each of three polyamines, results in an increase also of the 5' nucleotidase and adenosine deaminase activity. PMID- 6279450 TI - Mechanism and contribution of the pentose phosphate cycle to glucose metabolism in epididymal fat tissue. AB - 1. Glucose 6-phosphate, fructose 6-phosphate and altroheptulose 7-phosphate are the major products formed non-oxidatively from ribose 5-phosphate by rat epididymal fat pad enzymes. 2. Arabinose 5-phosphate was detected among the reaction products and significant activity of the new enzyme of the L-type pentose pathway, D-glycero D-ido octulose 1,8-bisphosphate: D-altroheptulose 7 phosphotransferase was found. 3. The glucose moieties of glucose 1-phosphate, glucose 6-phosphate and glucose 1,6-bisphosphate were degraded and showed that epididymal fat pad enzymes relocate 14C from [2-14C]glucose into C-1, C-2, and C 3 of each hexose-phosphate. 4. The 14C-distribution patterns in the hexose phosphates revealed that these intermediates were not in isotopic equilibrium and the rate of the transaldolase exchange reaction was relatively small. 5. The 14C distribution data suggest that glucose 1-phosphate, rather than glucose 6 phosphate, is the first intermediate in the path of glycogen synthesis from glucose in this tissue. 6. The data provide the first proof of the mechanism of the pentose pathway in adipose tissue. PMID- 6279452 TI - Multiple forms and inhibitors of uridine-cytidine kinase in neoplastic cells. AB - 1. Two forms (isozymes) of uridine (urd)-cytidine (cyd) kinase are present in the 30-50% ammonium sulfate fraction of the cytosols of L1210 ascites leukemia cells and a human malignant lymphoma. 2. These findings confirm those which described multiple forms of urd-cyd kinase in tumors with rapid growth rate. 3. Studied of inhibitors (nucleoside analogs) of urd-cyd kinase derived from L1210 and 6410 leukemia cells resulted in the finding of four possible inhibitors of this enzyme. PMID- 6279453 TI - Chemical reactivity of oxygen-derived radicals with reference to biological systems. PMID- 6279454 TI - Regulation of manganese superoxide dismutase by oxygen in Saccharomyces cerevisiae. PMID- 6279455 TI - Progestins inhibit FSH-induced functional LH receptors in cultured rat granulosa cells. AB - The role of intraovarian progesterone in the control of follicular growth and development remains unclear. The presence of a rat ovary granulosa cell progesterone receptor suggests that progesterone has a direct effect on the follicles. We have previously reported that progestins inhibit FSH-stimulated estrogen production by cultured granulosa cells by inhibiting the FSH induction of the aromatase enzyme. We now report that progestins can inhibit another FSH action on rat granulosa cells; the induction of LH/hCG receptors. The concomitant administration of 10(-5) M R5020, a potent synthetic progestin, with 10 ng/ml FSH during a 2-day culture period inhibits the FSH induction of LH/hCG receptors by 75 +/- 6% (mean +/- S.E.) The progestin inhibition of the induction of LH/hCG receptors is not mediated by its inhibitory action on the induction of aromatase. Scatchard analysis indicates that progestin decreases the number of LH/hCG receptors per cell but has no effect on receptor affinity. Both R5020 and progesterone have a dose-dependent inhibitory effect on the FSH induction of LH/hCG receptors, causing a 30 and 85% decrease in receptor number at concentrations of 10(-6) and 10(-5) M, respectively. The concomitant administration of R5020 with FSH also leads to a significant decrease in the ability of LH to stimulate cAMP production, indicating that progestin is inhibiting the induction of 'functional' LH/hCG receptors. R5020 (10(-5) M) also inhibits by 90% the induction of LH/hCG receptors by cholera toxin and dibutyryl cAmP, indicating that the progestin effect is at a post-cAMP site. Since the induction of LH/hCG receptors by FSH is a necessary event in follicular maturation, these results offer another mechanism by which progestins, at high concentrations, can inhibit follicular growth and development. PMID- 6279456 TI - Direct visualization of membrane clustering and endocytosis of thyrotropin into cultured thyroid cells. AB - We prepared a highly fluorescent conjugate of thyrotropin (TSH) which retains approximately 25% of the binding affinity of native TSH towards TSH receptor and approximately 25% of the potency of the native hormone in stimulating the accumulation of cAMP in thyroid cells. Using an image-intensified microscopy system, we observed that our fluorescent TSH bound specifically to diffusely distributed membrane receptors on live rat or bovine embryo thyroid cells grown in culture. At 37 degrees C the fluorescent hormone formed visible patches which were internalized and subsequently degraded. Hence, TSH, like other polypeptide hormones, is internalized by a process of receptor-mediated endocytosis. The addition of bivalent antibodies against the fluorophore rhodamine, to thyroid cells that were previously exposed to sub-optimal concentrations of rhodamine TSH, elicited maximal stimulation of the thyroid adenyl cyclase. Monovalent Fab' fragments did not enhance the response of rhodamine TSH. Therefore we conclude that enhanced surface clustering of TSH molecules increases their capacity to stimulate the adenyl cyclase of thyroid cells. PMID- 6279457 TI - The role of cyclic AMP in the induction of estrogen and progestin synthesis in cultured granulosa cells. AB - The role of cyclic AMP in the induction of enzymes involved in estrogen and progestin biosynthesis in undifferentiated granulosa cells was investigated. When granulosa cells from immature hypophysectomized, DES-treated rats were cultured for 2 days in serum-free medium with aromatase substrate (10(-7) M androstenedione) together with graded doses of FSH, prostaglandin E2 (PGE2), cholera toxin (CT), or dibutyryl cyclic AMP (Bu2cAmP), there was a dose-related increase in estrogen (E) production. The induction of E production by saturating doses of FSH, PGE2, CT, and Bu2cAmP required a lag phase of approximately 24 h, after which the E response increased sharply to maximum levels at day 3, and then declined gradually to day 5. Treatment for 24 h ((day 0-1) with FSH, together with 1 microgram/ml of either actinomycin D or cycloheximide, completely abolished the stimulatory action of FSH on E production. When the inhibitors were removed, the FSH-induced increases in E returned to near normal levels after a 24 h lag period. Similar effects of the inhibitors upon E production by CT, PGE2 and Bu2cAMP were observed. As with E, the production of progesterone and 20 alpha dihydroprogesterone was markedly stimulated by FSH, PGE2, CT and Bu2cAmP, and the results of the time course, dose response, and inhibitor experiments were similar to those for E production. These results indicate that FSH induces the de novo synthesis of enzymes required for both estrogen and progestin biosynthesis by undifferentiated granulosa cells and suggest that this action is mediated by cyclic AMP. PMID- 6279458 TI - Fructose 2,6-bisphosphate: a mediator of hormone action at the fructose 6 phosphate/fructose 1,6-bisphosphate substrate cycle. PMID- 6279459 TI - Non-specific uptake of the radioligand 125I-IHYP by intact human lymphocytes: reversal of the uptake process. AB - Binding of (+/-)[125I]iodohydroxybenzylpindolol (IHYP) to beta-adrenoceptors on intact human peripheral blood lymphocytes was found to be associated with a high degree of non-specific intracellular uptake of the radioligand. Non-specific binding on cellular surfaces was identified by displacing IHYP from its specific binding sites with the cold competing antagonist (+/-)-propranolol. Another methodological approach, however, was necessary to eliminate the uptake problem: release of intracellularly accumulated IHYP was achieved by sedimentation of the IHYP-loaded cells and resuspension in hypotonic buffer (HME buffer) for 10 min at +4 degrees C. The true value of maximal IHYP binding measured after IHYP release was 970 binding sites/cell as compared with 2300 receptors/cell found under standard binding conditions. This difference in maximal IHYP binding obtained by the 2 methods is attributed to the strong interference of the uptake process with the measurement of specific IHYP binding. Further evidence for the efficiency of the procedure to reverse the uptake process was obtained with the very slowly dissociable beta-adrenoceptor antagonist FM 24. Pre-incubation of the cells with 5 x 10(-5) M FM 24 totally prevented specific IHYP binding. Any IHYP measured on cells in the presence of FM 24, therefore, must have been due to mere non specific binding and cellular uptake of the radioligand. Even though the uptake of IHYP was enhanced in FM-24-treated cells, this influx could be completely reversed by exposing the IHYP-loaded cells to HME buffer for 10 min at +4 degrees C. Our findings indicate that our experimental conditions are suitable for beta receptor binding studies on intact cells with IHYP, because the uptake of the labeled ligand is eliminated. PMID- 6279460 TI - Inhibition of FSH action on granulosa cells by low molecular weight components of follicular fluid. AB - Immature female rats were injected with a single dose (10 IU) of pregnant mare serum gonadotropin to induce growth and maturation of ovarian follicles. Using such ovaries as a model, we tested the effects of low molecular weight subfractions of charcoal-absorbed bovine follicular fluid (FF-c) on (a) radioiodinated human FSH (125I-hFSH) binding to ovarian homogenates, (b) ovine FSH-stimulated adenylate cyclase activity in granulosa cell homogenates and (c) cAMP production by intact granulosa cells. The follicular fluid was fractionated by ultrafiltration through membranes of differing pore-sized into molecular weight components of 1000-5000 (passing Amicon H1P-5 hollow fibers but retained by Amicon UM-2 membrane) and 500-1000 (passing Amicon UM-2 membrane but retained by Amicon Um-05 membrane). These low molecular weight fractions inhibited 125I hFSH binding to ovarian receptors, FSH-stimulated cAMP production by rat granulosa cells and FSH-stimulated, as well as fluoride-ion-stimulated adenylate cyclase activity in granulosa cell homogenates. Inhibition of FSH-stimulated adenylate cyclase activity by the FF subfractions was non-competitive as determined by double reciprocal plot analysis. Our results suggest that modulation of FSH effects on granulosa cells may be mediated by low molecular weight constituents of follicular fluid. PMID- 6279461 TI - Characterization of solubilized prolactin receptors from Rana catesbeiana tadpole tissues. AB - Prolactin (PRL) receptors were solubilized from Rana catesbeiana tadpole liver and tail fin and female Sprague-Dawley rat-liver membranes by treatment with 1% Triton X-100. Binding of [125I]oPRL to tadpole-liver and tail-fin solubilized extracts reached equilibrium by 12 h at 19 degrees C. The binding was linear up to 50 micrograms of tadpole liver and tail-fin protein and 30-40 micrograms of rat-liver protein. Solubilization did not affect the dissociation constant for [125I]oPRL binding but did result in a greater number of sites. The binding was specific for oPRL and hGH, which has PRL-like activity. Neuraminidase pretreatment of membranes altered the binding affinity of oPRL to tadpole-liver membranes and solubilized extracts but not to tadpole-tail fin membranes or extracts. Pretreatment of membranes with neuraminidase did not affect the binding capacity of tadpole-liver or tail-fin membranes or solubilized extracts. PMID- 6279462 TI - Regulation of rat pineal melatonin synthesis: effect of monoamine oxidase inhibition. AB - Inhibition of pineal monoamine oxidase (MAO) activity either by harmine or pargyline in adult male Sprague-Dawley rats housed in a 12 : 12 LD cycle resulted in increase pineal N-acetyltransferase (NAT) activity. Pineal MAO inhibition also increased pineal melatonin content, presumably as a result of the increased NAT activity. Conjunct treatment with propranolol, a beta-adrenergic receptor antagonist, nullified these effects, regardless of the MAO inhibitor (harmine, pargyline or both) used or the inhibitor dose given. MAO inhibition during continuous light resulted in increased NAT activity greater than that observed following MAO inhibition during a 12 : 12 LD cycle. On the other hand, the increase in melatonin content following MAO inhibition during continuous light was not significantly different from that following MAO inhibition during a 12 : 12 LD cycle. Conjunct propranolol administration negated the effects of MAO inhibition on both the level of NAT activity and melatonin content, regardless of the lighting conditions. The level of pineal NAT activity is apparently regulated by the level of pineal beta-adrenergic receptor stimulation. While melatonin production appears to be dependent on increases in NAT activity, biosynthesis of this methoxyindole may also be regulated, in part, by other factors or processes in metabolic pathway. PMID- 6279463 TI - Expression of a phenotype of normal differentiation in cultured mammary glands is promoted by epidermal growth factor and blocked by cyclic adenine nucleotide and prostaglandins. PMID- 6279464 TI - Enhancement of squamous cell development in cultured skin by cyclic adenine nucleotide and prostaglandins. AB - The present study tests the hypothesis that agents known to elevate the level of intracellular cyclic adenine nucleotide may direct different epithelial cells onto a pathway of epidermoid (squamous) development and differentiation. We report here that the mixture of dibutyryl cyclic AMP (dbcAMP), prostaglandins E1, E2 and B1 (PG E1, E2, B1), and papaverine (pap) enhances the rate of normal squamous cell development in organ-cultured skin of chick embryos. The three components may act synergistically to elevate the level of intracellular cyclic adenine nucleotide. We recently reported that the same group of agents induces abnormal development (squamous metaplasia) and aberrant differentiation (keratin production) in the normally cuboidal epithelium of cultured whole mammary glands of mice [1]. Thus, dbcAMP, PG E1, E2, B1, and pap are effective in enhancing normal squamous cell development and also in inducing squamous metaplasia de novo in the epithelial components of two different organs of embryonic and adult animals of two classes of vertebrates. The combined findings are suggestive that cyclic adenine nucleotide together with the prostaglandins may act generally on diverse types of epithelia to bring about squamous cell development and a differentiation marked by keratin production. PMID- 6279466 TI - Inhibition of basal acid, chloride, and pepsin secretion in duodenal ulcer by graded doses of ranitidine and atropine with studies of pharmacokinetics of ranitidine. PMID- 6279465 TI - Effect of gastrointestinal hormones on the biliary sphincter of the opossum. AB - The smooth muscle sphincter enveloping the terminal portion of the common bile duct in the opossum exhibits spontaneous electrical activity and simultaneous rhythmic contractions. The aim of our study was to define the influence of four gastrointestinal hormones on biliary sphincter electrical and mechanical activity. An array of five monopolar extracellular electrodes was placed along the opossum choledochal sphincteric smooth muscle and contiguous duodenum. A catheter in continuity with a pressure transducer, drop counter, and saline reservoir was placed in the common duct for simultaneous measurement of ductal pressure and flow. The cystic and distal common hepatic ducts were then ligated to isolate the common bile duct from the gallbladder and liver. In each opossum, biliary sphincteric and duodenal myoelectric activity, common bile duct and gallbladder pressure, and common duct flow were recorded simultaneously before and after the intravenous administration of five different doses of an enteric hormone. Ten animals were given 0.1-10.0 international dog units per kilogram body wt of cholecystokinin, 10 received 0.01-1.00 microgram/kg body wt of cholecystokinin-octapeptide, 10 were given 0.1-10.0 micrograms/kg body wt of secretin, and 5 were given 0.1-10.0 micrograms/kg body wt of pentagastrin. Cholecystokinin, cholecystokinin-octapeptide, and pentagastrin all effected a significant increase in sphincter electrical spike activity and common duct pressure with a decrease in common duct flow. This contractile response was consistent at a wide range of hormonal levels. Secretin had little effect on biliary pressure, flow, and myoelectric activity. The data lend support to the concept that cholecystokinin and gastrin contract the biliary sphincter, metering bile flow at the time of gallbladder emptying in the opossum. PMID- 6279468 TI - The binding of pancreatic hormones to isolated chicken hepatocytes. PMID- 6279467 TI - Quantification of the fate of dietary fiber in humans by a newly developed radiolabeled fiber marker. AB - A radiolabeled cellulose (131I-fiber) that retains the essential physical and chemical properties of this class of fiber was developed in our laboratory. We quantified the fate of orally ingested 131I-fiber in healthy individuals by external gamma camera monitoring and fecal collections. The marker passes virtually intact through the human gastrointestinal tract with negligible release and absorption of the label in the gut. Comparison of the gastric emptying rate of 131I-fiber with that of a predominantly aqueous marker, 99mTc diethylenetriamine pentaacetic acid (99mTc-DTPA), showed that 131I-fiber strands were evacuated more slowly than intragastric fluids. An important finding was that some 131I-fiber emptying occurred during most time periods, even before liquids were completely evacuated. This suggests that the human stomach is able to empty simultaneously liquids and fiber strands (1-15 mm in length) that are resistant to grinding by antral mechanical forces and to digestion by acid-peptic secretion. Thus, some nondigestible solids may be emptied with the bulk of a meal, although at a slower rate. 131I-Fiber may be a useful marker for quantifying gastric emptying of nondigestible solids. Further, the stability of 131I-fiber in the gut, as opposed to most other physiologic solid labels, should enable future investigation of intestinal and colonic transit of fiber, which is an important component of the human diet. PMID- 6279469 TI - Hormonal effects on cyclic nucleotides and carbohydrate and lipid metabolism in isolated chicken hepatocytes. PMID- 6279471 TI - Hypoglycemic coma: don't overlook acute adrenal crisis. PMID- 6279470 TI - [Ultrastructure of different cells of the adenohypophysis of the Saharian lizard Uromastix acanthinuris Bell 1825 (Sauria Agamidae)]. PMID- 6279472 TI - [Effect of ultraviolet radiation on the electrochemical potential and fibrogenic properties of quartz dust]. PMID- 6279473 TI - [Occupational hygiene in the production of potassium fertilizers]. PMID- 6279474 TI - Health fairs for older adults. After the fair is over... PMID- 6279475 TI - [Behaviour of two second messengers; cAMP and cGMP in leucocytes of peripheral blood of normal and leukemic subjects (author's transl)]. PMID- 6279476 TI - Platelet aggregation and thromboxane A2 production after adrenergic stimulation in young healthy humans. AB - The effects of adrenergic stimulation on platelet aggregation (platelet aggregation ratio; PAR), beta-thromboglobulin (beta-TG) release and plasma thromboxane B2 (TxB2) levels were investigated in 25 healthy young volunteers. Adrenergic stimulation induced by cold application was checked by evaluating the changes in the calculated vascular resistance in the forearm. A prompt increase in platelet aggregates and plasma beta-TG and TxB2 concentrations was observed after adrenergic stimulation. PAR changed from resting values of 0.97 +/- 0.05 to 0.75 +/- 0.08 (p less than 0.001) at the end of cold application. At the same time, beta-TG plasma concentration increased from 32.09 +/- 19.64 to 135.48 +/- 37.97 ng/ml (p less than 0.001) and TxB2 plasma levels changed from 0.49 +/- 0.24 to 0.99 +/- 0.39 pmol/ml (p less than 0.001). TxB2 levels, but not PAR and beta TG concentration came back to the resting values at the end of the observation period (10 min). Aspirin, as the lysine acetylsalicylate equivalent to 5 mg/kg i.v. of acetylsalicylic acid, although able to completely inhibit platelet cyclooxygenase failed to inhibit the plasma TxB2 increase induced by adrenergic stimulation. This strongly suggests that the increase in plasma TxB2 following adrenergic stimulation is of extraplatelet origin. Also beta-TG and PAR changes were not affected by aspirin administration. PMID- 6279478 TI - Glucagon receptors: are they present on human blood cells? AB - Binding of glucagon to and its degradation by human mononuclear leucocytes and erythrocytes was investigated. Labeled glucagon binds to these cells, but can only be displaced by unlabeled glucagon at pharmacologically high concentrations. It is concluded that glucagon binding to human blood cells does not represent receptors of physiological importance and is of no value for the determination of glucagon sensitivity in man. PMID- 6279477 TI - [Lithium as a psychotropic drug. Experimental studies on its mechanism of action]. AB - The effect of lithium on the cAMP metabolism was studied in rat brain. It was shown that: 1. Acute and subacute doses of lithium have no effect on the cAMP content of the forebrain of rats while chronic administrations significantly reduce the cAMP level in the brain by 30%; 2. the activities of the cerebral cAMP and cGMP phosphodiesterases are not affected by lithium in vitro; 3. the adenyl cyclase in rat brain homogenates stimulated by sodium fluoride and noradrenaline is inhibited dose-dependently in the mM range and 4. adenyl cyclase in isolated cells from the rat forebrain is very sensitive to inhibition by lithium in the therapeutically relevant microM range. With all due caution in transferring experimental results to the mechanisms of action of drugs in humans, it is obvious from these results in comparison with the effect of lithium on hormone sensitive adenyl cyclases from extracerebral tissues that the effect of lithium salts in manic psychoses may be due to inhibition, and their prophylactic action probably to a "stabilizing" of cerebral adenyl cyclases. PMID- 6279480 TI - Effect of adrenalectomy on glucagon binding in rat fat cells. PMID- 6279479 TI - Binding of a variant of human growth hormone to liver plasma membranes. AB - The binding of human growth hormone (hGH) and a naturally occurring variant of hGH (20K) to rat and rabbit liver membrane has been compared. The concentration of hormone causing half maximal displacement (ED50) of 125I-hGH from rat liver plasma membranes was 1.6 nM for hGH and 46 nM for 20K. The ED50 for hGH and 20 K against 125I-hGH binding to rabbit liver plasma membranes was 1.6 mM for hGH and 28 nM for 20K. Significant binding of 125I-20K did occur to rat and rabbit liver membranes but the relative capacities for 20K were 12 and 6 fold less in rat and rabbit liver membranes, respectively when compared to hGH. The competitive binding characteristics of 20K to pregnant rabbit liver membranes indicated lack of competition between bovine GH and 125I-20K for binding. However, significant binding of 125I-20K to male rabbit liver membranes was present and displaceable by hGH and oPrl. We conclude that deletion of a portion of the hGH molecule (defined as residues 32-46) results in significant alterations in the binding characteristics of the molecule. This indicates primary or secondary involvement of this portion of the hGH molecules in the binding process and may define a portion of the binding domain for hGH. PMID- 6279481 TI - Dexamethasone-suppressible hypercorticosteronism in two 46,XX subjects with ambiguous genitalia and ovarian cysts. Partial defect of 17 alpha-hydroxylase or 17-20-desmolase. AB - The paradoxical association of female pseudohermaphroditism and androgen deficiency was observed in two 46,XX subjects with high corticosterone plasma levels. Subject 1 has been declared a boy due to clitoris enlargement; she had no vagina and uterus. Subject 2 had ambiguous external genitalia. In both, at age 27 and 17 years, fusion of outer labia, impuberism, ovarian cysts, and histologically normal ovarian tissue were observed. Blood pressure was normal. Basal cortisol levels were normal but unresponsive to ACTH. Progesterone levels were 40 and 62 ng/ml and rose after ACTH (50 and 79 ng/ml). 17 hydroxyprogesterone levels were 25 and 21 ng/ml and did not rise after ACTH. Corticosterone levels were 70 and 92 ng/ml and rose after ACTH (110 and 180 ng/ml). All three steroids were suppressed by dexamethasone. Androgen and estrogen levels were at or below the lower limit for normal women. The sex steroid levels obtained by radioimmunoassay in plasma and a follicular cyst fluid were confirmed by isotope dilution-mass spectrometry. We suggest that the sexual ambiguousness resulted from an excessive production of gestagenic steroids during fetal life, and that the enzyme defect is either a partial 17 alpha-hydroxylase defect combined with a peripheral production of 17-hydroxyprogesterone, or else a partial 17-20-desmolase defect with a secondary 21-hydroxylase defect limited to the cortisol pathway. PMID- 6279482 TI - Responsiveness of cortisol and dehydroepiandrosterone to ACTH in children. AB - In a total of 101 children, the dehydroepiandrosterone (DHA) and cortisol (F) levels were measured before and after ACTH (Synacten) administration. F responsiveness was unchanged during development, while DHA responsiveness in healthy children was highest during adrenarche. In hypopituitary patients DHA levels were lower than in the controls, but responsiveness to ACTH showed similar changes during development. Children with Turner's syndrome and hypergonadotrophic males had the response in elevated DHA levels while ACTH induced DHA response related to bone-age matched controls. We conclude that regulation of adrenal androgens is mediated by both ACTH and another hypothalamo pituitary hormone, perhaps independent of gonadal activation, but requiring gonadal integrity. PMID- 6279483 TI - Concanavalin A reduces liver collagen accumulation in murine schistosomiasis. AB - Mice infected with Schistosoma mansoni develop hepatic fibrosis associated with enhanced collagen synthesis that out-paces induced collagenase activity. Administration of one dose of concanavalin A [Con A (200 micrograms)] by i.p. injection to mice at 5 or 6 weeks after infection with 50 S. mansoni cercariae decreased liver collagen content by 50% compared to levels in control-infected mice injected with either homologous immunoglobulin (200 micrograms) or phosphate buffered saline; additional doses of Con A had no further effect. The decrease in collagen content could not be attributed to either decreased egg deposition in the liver or inhibition of liver collagen synthesis, but was coincident with a greater solubility of granulomas. Collagen contents of skin and tail were unaffected. The relative solubilities of liver collagen in 8 M urea: 10 mM dithiothreitol were greater in treated animals as compared to controls. However, the amounts of collagen solubilized were similar in both sets of animals, since the total collagen content of treated mice was 50% of the controls. A possible explanation for these results is that much of the synthesized collagen does not accumulate in treated animals, whereas it does accumulate in controls. Peak collagenase and neutral protease activities occurred at 7 weeks postinfection in treated animals, and were 2-fold greater than in controls. Similar effects were observed when succinylated Con A was administered. The results suggest that Con A may modulate host-immune responses influencing fibrogenesis in hepatic murine schistosomiasis. PMID- 6279484 TI - Hepatic catabolism of low density lipoprotein: mechanisms and metabolic consequences. PMID- 6279486 TI - Epinephrine-induced conditioned taste aversion. PMID- 6279485 TI - Liver and cellular vitamin A binding proteins. PMID- 6279487 TI - Polymorphism of human galactose-1-phosphate uridyl transferase. AB - Frequencies for the electrophoretically detectable alleles of galactose-1 phosphate uridyl transferase were determined from 406 neonates in Adelaide. GALT*D and GALT*LA frequencies were 0.054 and 0.027, respectively. The expected proportion of heterozygotes was 15.3%, with a GALT N-D component of 10%. The distinction between GALT*D and GALT*LA is essential for estimation of the expected frequency of GALT DG genotypes, an unknown proportion of which are detected as 'deficient' from neonates on galactosaemia screen. PMID- 6279488 TI - Generation of cross-reactive cytotoxic T lymphocytes following immunization of mice with various bluetongue virus types. AB - Mice immunized with a single bluetongue (BT) virus type were shown to produce cytotoxic T lymphocytes (CTL's) which cross-reactive with a number of BT virus types. These cross-reactive CTL's could be induced by both primary in vivo and secondary in vitro stimulation. A varying degree of cross-reactivity occurred with the six BT types examined. Aspects of the character of this cross-reactivity were examined and its role in protection from disease and vaccination strategy is discussed. PMID- 6279489 TI - Differences in the ingestion mechanisms of IgG and C3b particles in phagocytosis by neutrophils. PMID- 6279490 TI - Split T-cell tolerance in herpes simplex virus-infected mice and its implication for anti-viral immunity. AB - Mice simultaneously injected intravenously and subcutaneously with herpes simplex virus fail to adoptively transfer delayed hypersensitivity (DH) to syngeneic recipients. The transferred lymph node cells also failed to rapidly eliminate infectious herpes from the pinna, despite the presence of cytotoxic T cells in the transferred suspension. Both primary and secondary cytotoxic cell responses in the draining lymph node were unaffected by the inhibition of DH. The lymph nodes from DH tolerized mice also contain lymphocytes capable of undergoing a proliferative response in vitro to herpes antigens. In addition, a neutralizing antibody response with IgG antibodies against herpes are also present in DH tolerized mice. These data suggest a form of split T-cell tolerance in which only DH responses are directly compromised. The implication of these findings for the pathogenesis of herpes simplex virus is discussed. PMID- 6279491 TI - Alpha-adrenoceptors in the rabbit intestine and aorta. PMID- 6279492 TI - Clostridium perfringens in faeces of healthy persons and diarrhoeal patients. PMID- 6279493 TI - Acute hypertension in a nonhuman primate: humoral and hemodynamic mechanisms. AB - The present study assessed the contribution of the renin-angiotensin system (RAS), dietary sodium, and cardiac output (CO) to the genesis of primate hypertensin in a one-kidney model which was developed to test species-specific renin inhibitors. Reduction of renal perfusion pressure increased mean arterial pressure (MAP) from 105 +/- 4 to 127 +/- 3 mm Hg (p less than 0.0005), associated with increased plasma renin activity (PRA) from 4.9 +/- 0.7 to 13.8 +/- 1.1 ng. ml-1.hr-1 (p less than 0.0005). Correlation of MAP with PRA yielded an r value of 0.662 (p less than 0.0005). Significant blood pressure elevation was obtained with both regular (R) and low sodium (LS) diet (p less than 0.0005), although the MAP change was greater with LS. With both R and LS diet, hypertension was associated with increased PRA (p less than 0.0005), and normotensive pressures were achieved with converting enzyme inhibitor (teprotide). The hemodynamic change with hypertension was an increase of systemic vascular resistance (SVR) from 0.89 +/- 0.12 to 1.17 +/- 0.09 units (p less than 0.05); cardiac output (CO) and central blood volume did not change significantly. Thus, acute hypertension, mediated by the RAS, was developed in a one-kidney primate model. The hemodynamic correlate of hypertension was increased SVR; CO and volume redistribution were not initiating factors. PMID- 6279494 TI - Responsiveness to pressor agents in experimental renovascular and steroid hypertension. Effects of converting enzyme inhibitor and nephrectomy. PMID- 6279495 TI - Increased circulating levels of an endogenous digoxin-like factor in hypertensive monkeys. AB - An endogenous, immunoreactive digoxin-like factor (endoxin) was measured in the plasma of nonhuman primates with hypertension. Both normotensive and hypertensive rhesus monkeys had levels of endoxin that significantly correlated with their systolic or diastolic blood pressure. Vervet monkeys with experimentally produced chronic Goldblatt hypertension had significantly elevated endoxin, but not plasma renin. These data suggest that increased plasma endoxin may be a contributing factor in the development of hypertension. PMID- 6279496 TI - Aldosterone infusion into the rat and dose-dependent changes in blood pressure and arterial ionic transport. AB - Induction of hypertension by implantation or injection of deoxycorticosterone acetate (DOCA) requires a dose well above the physiological range. The objective of this study was to produce hypertension in rats by chronic infusion of d aldosterone, a more potent mineralocorticoid. Aldosterone infused at a dose of 1 microgram/hr for 4 weeks gave maximal rise in systolic blood pressure (132 +/- 3 vs 203 +/- 7 mm Hg). A significant rise in blood pressure was achieved at 0.1 microgram/hr (170 +/- 6 mm Hg) which was associated with a 2.3-fold rise in plasma aldosterone levels (7.6 +/- 0.4 vs 17.7 +/- 2.2 ng/dl). A series of isotope flux studies on the aorta and femoral artery from hypertensive animals demonstrated increases in 42K and 36Cl turnover. In both vessels the largest changes in ion turnover were observed in vessels from animals infused with aldosterone at 0.25 micrograms/hr. Increases in 42K and 36Cl turnover were detected as early as 1 week after the start of aldosterone infusion, well before a significant rise in blood pressure had occurred. PMID- 6279497 TI - Function of the sodium pump in arterial smooth muscle in experimental hypertension: role of pressure. PMID- 6279499 TI - Chronic potentiation of vasoconstrictor hypertension by adrenocorticotropic hormone. AB - The chronic effects of ACTH on mean arterial pressure (MAP) and related variables were studied in dogs with both chronic norepinephrine (NE)- and chronic aldosterone-induced hypertension. MAP was recorded continuously for 24 hours/day, and sodium intake was 71 mEq/day. ACTH was infused for 8 days at a rate that does not increase MAP in normotensive dogs and yet a rate that produces pronounced mineralocorticoid and glucocorticoid effects. Chronic ACTH infusion in dogs with NE hypertension caused natriuresis, kaliuresis, diuresis, hypernatremia, hypokalemia, and suppression of PRA; additionally, there was either no net change in water balance or net water balance was positive. However, in marked contrast to dogs without pre-existing hypertension, in dogs with NE hypertension ACTH produced a pronounced additional increase in MAP of 39 to 63 mm Hg. Although ACTH markedly potentiated NE hypertension, high infusion rates of aldosterone (+6 mm Hg) and cortisol (-7 mm Hg) had relatively weak effects on MAP; further, in dogs with NE hypertension, the increase in MAP associated with simultaneous infusions of high rates of cortisol and aldosterone was equal to only approximately half of that produced by ACTH. In dogs with aldosterone hypertension, the changes in salt and water balance produced by ACTH were comparable to those that occurred when ACTH was administered to dogs with NE hypertension. In dogs with aldosterone hypertension, however, ACTH did not produce kaliuresis, hypernatremia, or hypokalemia; moreover, ACTH did not exacerbate aldosterone hypertension. Thus, the data indicate that the hypertensive effects of ACTH are manifested in conditions of reduced renal excretory capacity such as exist when plasma levels of the potent sodium -retaining hormone NE are inappropriately elevated. Finally, the hypertensive effects of ACTH cannot be accounted for simply on the basis of enhanced mineralocorticoid and glucocorticoid activity. PMID- 6279498 TI - Dopaminergic modulation of pressor and hormonal responses in essential hypertension. AB - Hormonal and mean arterial pressure (MAP) responses to posture, isometric handgrip, angiotensin II (AII), adrenocorticotrophic hormone (ACTH), and metoclopramide (MCP), a dopamine (DA) antagonist, were examined in nine men with essential hypertension and nine age- and weight-matched normotensive men on a constant 100 mEq sodium and 80 mEq potassium intake before and after 4 days of administration of the DA agonist, bromocriptine (BEC; 2.5 mg three times a day). BEC depressed supine basal MAP in the hypertensives, and decreased MAP response to posture and isometric exercise in both groups. Hypertensives displayed greater (p less than 0.01) NE responses to posture and exercise than the normotensives. BEC decreased the NE response to 10 minutes of upright posture and exercise more in hypertensives (p less than 0.01) than in normotensives, but following BEC, the responses were similar. BEC did not affect basal PRA or PRA responses to posture and exercise in the two groups. PA responses to ACTH and MCP were similar in both groups, but the hypertensives displayed greater (p less than 0.01) PA responses to AII. BEC suppressed PA responses to AII (p less than 0.01) and to high dose ACTH (p less than 0.05) to a similar extent in both groups. The prolactin as well as the PA response to DA antagonism with MCP was similar in the two groups. These results suggest that dopaminergic control of NE secretion may be altered in essential hypertension. Blood pressure lowering effects of BEC in patients with essential hypertension may be related, in part, to depression of sympathetic nervous system activity. PMID- 6279500 TI - Alterations in osmotic but not pressor responses to ACTH by optic recess lesions in sheep. AB - This study examines whether neural structures in the region of the optic recess of the third ventricle may be involved in the genesis of adrenocorticotrophic hormone (ACTH)-induced hypertension in sheep. Five sheep were prepared with lesions in an area of the forebrain that included the organum vasculosum of the lamina terminalis (OVLT) and surrounding periventricular tissue. In these animals the dipsogenic response to systemically infused hypertonic sodium chloride (NaCl) was abolished. ACTH treatment (20 micrograms/kg/day) for 5 days caused an increase in mean arterial pressure (MAP) of 19 mm Hg, a response identical to that seen in normal sheep. With ACTH treatment, increases in plasma osmolality were greater than normal, but polydipsia did not occur in the lesioned sheep. In six other sheep with lesions either lateral or anterior to the optic recess of the third ventricle, the dipsogenic response to hypertonic NaCl and pressor response to ACTH were normal. These studies establish that in ACTH-treated sheep the integrity of the anterior ventral part of the third ventricle is not essential for the development of the hypertension. This is in contrast to the finding in other models of experimental hypertension in the rat. PMID- 6279501 TI - Antibodies to the alpha 1- and alpha 2-selective antagonists prazosin and yohimbine as probes of the alpha-adrenergic binding sites. AB - Antibodies were raised against a newly synthesized analog (CP57,609) of the alpha 1-selective antagonist prazosin, and against the alpha 2-selective antagonist, yohimbine, by immunization of rabbits with antigens prepared by covalent linkage of these ligands to albumin. Competitive inhibition of [3H]prazosin binding to anti-CP57,609 antiserum by a variety of unlabeled ligands revealed a spectrum of antibody specificity, with alpha 1-selective agents competing more potently than alpha 2-selective ligands. In contrast, alpha 2-selective ligands competed more potently with the binding of [3H]yohimbine to the anti-yohimbine antiserum than alpha 1-selective agents. These respective antisera were subjected to affinity fractionation of a CP57,609- or yohimbine-Sepharose 4B resin. Fractions from the CP57,609 resin were eluted successively with phentolamine (10(-3)M), prazosin (10(-4)M), and guanidine (5M), and from the yohimbine resin with prazosin (10( 4)M), yohimbine (10(-4)M), and guanidine (5M). The binding profiles of these fractions differed, and in certain fractions the relative order of potency of adrenergic agents was almost identical to that observed with membrane alpha adrenergic receptors. Moreover, using these eluted fractions as immunogens, antisera have been obtained which, in the initial bleeds, already possess antiidiotypic activity. These findings therefore suggest that affinity fractionation of antibodies raised against alpha 1- and alpha 2-selective antagonists may provide useful analogs for the further study of the ligand recognition properties of alpha-adrenergic receptors. Additionally, it is probable that antiidiotypic antisera will be developed which will recognize the alpha-adrenergic binding sites. PMID- 6279502 TI - Altered renal alpha 2-adrenergic receptor regulation in genetically hypertensive rats. AB - Renal alpha 1 and alpha 2-adrenergic receptors were quantified in Dahl salt sensitive and salt-resistant rats, in Okamoto-Aoki spontaneously hypertensive rats (SHR), in Wistar Kyoto "normotensive" (WKY), and in Charles River rats made hypertensive by the Grollman ligature technique and by DOC-NaCl administration after unilateral nephrectomy. The effect of high dietary NaCl on renal alpha receptors was studied in Dahl, SHR, and WKY rats. Renal alpha 1 and alpha 2 receptor densities were higher (p less than 0.05) in SHR and in Dahl salt sensitive rats than in their normotensive controls. High dietary sodium increased renal alpha 2 receptors and blood pressure in SHR, WKY and Dahl salt-sensitive, but not in resistant Dahl rats. A study of time relationships revealed that the increase in renal alpha 2 receptors preceded most of the blood pressure elevation due to high dietary sodium. Renal alpha-adrenergic receptor densities of surgical (Grollman) and endocrine (DOC-NaCl) forms of rat hypertension were not different from normotensive controls. Thus, renal alpha 2 receptor density and increase thereof by dietary sodium may be: 1) a biochemical marker for genetic forms of hypertension in the rat, and 2) closely linked to the basic mechanism of high blood pressure. PMID- 6279504 TI - Enhanced vasodilatation in essential hypertension by calcium channel blockade with verapamil. AB - The dependency of arteriolar tone on calcium influx was studied in 11 patients with essential hypertension (EH) and compared to 11 age-matched normotensive subjects (NT) by measuring the forearm blood flow response to intraarterial infusion of the calcium channel blocker verapamil (Verap) and the non-specific vasodilator sodium nitroprusside (Nip) using venous occlusion plethysmography. Verap in incremental dosages from 1 to 75 micrograms/100 ml forearm tissue induced a greater increase in forearm blood flow ( delta FAF) in EH then in NT, whereas there was no significant difference in delta FAF following Nip 1,2 micrograms/100 ml tissue. Delta FAF to Verap as adjusted for delta FAF to Nip was still greater in EH than in NT. Delta FAF to all dosages of Verap correlated positively with basal plasma epinephrine concentration in EH. At the two highest dosages of Verap, systemic blood pressure fell in EH, and the Verap-induced vasodilator (as adjusted for the response to Nip) correlated negatively to plasma renin activity or plasma angiotensin II concentration. These findings support the concept of an increased dependency of arteriolar tone on calcium influx in EH, which is related to the activity of the sympathetic nervous system. This association may be due to a common underlying derangement in transmembranous ionic fluxes in smooth muscle cells and sympathetic neurons in EH. PMID- 6279503 TI - Calcium- and endothelial-mediated vascular smooth muscle relaxation in rabbit aorta. AB - The role of calcium in the relaxations evoked by methacholine and A23187 in intact rabbit aortic rings was investigated. Methacholine (10(-8) to 10(-6) M) and the calcium ionophore A23187 (10(-8) to 10(-6) M) produced dose-dependent relaxations of rings which had been contracted with the alpha-adrenergic agonist phenylephrine. The ability of a ring to relax in this manner was correlated with the presence of endothelium as judged by transmission and scanning electron microscopy. Purposely disrupting the endothelium led to a loss of the relaxation response. In these rings methacholine caused dose-dependent contractions at concentrations greater than 10(-7) M. Deletion of Ca++ from the incubation medium inhibited maximum methacholine-induced relaxations by 67% and A23187-induced relaxations by 92%. The Ca++-channel blockers verapamil (10 microM) and nifedipine (0.5 microM) inhibited maximum methacholine-induced relaxations by 39% and 45%, respectively. The blockers had no effect on the methacholine ED50 (2.5 x 10(-7) M) for relaxation. Verapamil and nifedipine also inhibited maximum A23187 induced relaxations by 43% and 47% with no effect on the ED50 (6 x 19(-8) M) for relaxation. A structurally dissimilar vasodilator, sodium nitroprusside (10(-7) M), had no effect on the A23187-induced relaxation. These data are consistent with a role of Ca++ in regulating either the production or release of endothelial derived relaxing factor(s). PMID- 6279505 TI - Myosin phosphorylation and crossbridge regulation in arterial smooth muscle. State-or-the-art review. AB - Regulation of vascular resistance is generally explained in terms of neural, hormonal, metabolic, and myogenic factors altering intracellular calcium [Ca++] in vascular smooth muscle. Ca++ acts as a second messenger regulating the number of active crossbridges and force generation by binding to a myofilament regulatory protein. A search for the Ca++-binding regulatory protein in arterial smooth muscle has uncovered what appears to be a new type of regulation. In addition to its interaction with an undefined Ca++-binding site which determines force development, Ca++ stimulates phosphorylation of the crossbridges. Phosphorylated crossbridges cycle more rapidly than dephosphorylated crossbridges in the presence of Ca++. Some known characteristics of the myosin light chain kinase/phosphatase system and the effects of crossbridge phosphorylation on the mechanics of arterial smooth muscle are described. Chronic alterations in this system have potential effects on vascular resistance and merit investigation in studies of arterial smooth muscle from hypertensive animal models. PMID- 6279506 TI - Relative roles of sodium and calcium ions in the steroidogenic response of isolated rate adrenal glomerulosa cells. AB - To study the relative roles of sodium (Na+) and calcium ions (Ca2+) in the response of adrenal glomerulosa cells, we investigated the effects of different Na+ concentrations in the incubation media and the actions of substances that interfere with Ca2+ fluxes. Basal aldosterone secretion and response to angiotensin II (AII), adrenocorticotropic hormone (ACTH), or potassium (K+) were dependent on extracellular Na+ concentration. Veratridine, a Na+ channel opener that dissipates Na+ gradients, blocked the stimulated steroidogenic response. Mersalyl acid and tetracaine, which are potent Ca2+ antagonists, blocked the effects of aldosterone secretagogues. Divalent cations with Ca2+ antagonistic action such as manganese M(n2+), nickel (Ni2+), and cobalt (Co2+) blocked the aldosterone secretory response to AII, ACTH, and K+. Barium (Ba2+) and strontium (Sr2+), known to mimick Ca2+ effects, increased or did not affect responses of the glomerulosa cells. Sodium vanadate, an inhibitor of ATP-dependent Ca2+ translocation, did not alter the stimulated aldosterone responses. Trifluoperazine (10(-6) M), an inhibitor of calmodulin, blocked AII and K+ induced aldosterone secretion, but was partially effective on ACTH-stimulated aldosterone output only at a concentration of 10(-5) M. The actions of ouabain on aldosterone biosynthesis were similarly affected by all these drugs. Thus, both extracellular Na+ and Ca2+ appear to play a role in the steroidogenic response of isolated glomerulosa cells. The intracellular action of Ca2+ may involve a calmodulin-like protein. The effects of ACTH are only partially dependent on Ca2+ as a second intracellular messenger. PMID- 6279507 TI - Norepinephrine sensitivity and membrane potentials of caudal arterial muscle in DOCA-salt, Dahl, and SHR hypertension in rat. AB - Comparison of norepinephrine (NE) sensitivity in caudal arterial muscle of rats with three forms of hypertension showed that there was no increase in either DOCA salt or Dahl genetic hypertension, in contrast to the increased NE sensitivity found in spontaneously hypertensive rats (SHR). In hypertension induced by deoxycorticosterone acetate (DOCA)-salt treatment, as in Dahl genetic hypertension, there was also no difference in membrane potential (Em) between hypertensive and normotensive rats. By comparison to the SHR membrane alterations reported previously, any increased NE sensitivity might have been associated with altered Em electrogenesis which is triggered by a trophic factor of the sympathetic nervous system. SHR have a lower intracellular K+ free ion concentration and thus a smaller contribution of the ion gradient generated voltage which appears to be compensated for at rest by more active electrogenic ion transport. While SHR show greater depolarization and contraction than Wistar Kyoto (WKY) rats in response to midrange NE concentrations, DOCA-salt and Dahl hypertensive rat caudal arterial muscle showed neither NE hypersensitivity nor any evidence of altered Em mechanisms. Ion transport in isolated peripheral arteries in DOCA-salt hypertension may only secondarily be altered in response to primary changes in humoral factors and altered neural control mechanism. PMID- 6279508 TI - Aspects of angiotensin action in the adrenal. Key roles for calcium and phosphatidyl inositol. AB - The steps between exposure of bovine adrenal glomerulosa cells to angiotensin and the stimulated increase in aldosterone production were studied in two ways. Binding of angiotensin to receptors, and hormone effects on phosphatidyl inositol turnover, 45Ca2+ fluxes, and aldosterone production were measured directly. Other potential intermediate steps were investigated indirectly by use of inhibitors. Angiotensin slowed calcium influx and accelerated phosphatidyl inositol turnover in proportion to hormone dose. The effects correlated with receptor binding and aldosterone production. None of the inhibitors tested, except saralasin, inhibited angiotensin's effect on phosphatidyl inositol turnover. Altered calcium flux and stimulated aldosterone production were affected by the calmodulin inhibitor trifluoperazine and the intracellular calcium antagonist 8-(N,N diethylamino)-octyl 3,4,5-trimethoxybenzoate hydrochloride (TMB-8). Several reagents did not affect angiotensin binding, its effect on phosphatidyl inositol, or 45Ca2+ flux, but severely inhibited steroidogenesis. These included the phospholipase A2 inhibitor mepacrine, the protein synthesis inhibitor cycloheximide, and the Na+/k+-ATPase inhibitor ouabain. Colchicine had very little effect on the processes we measured, suggesting that microtubules play no role in angiotensin action in the adrenal. Based o these observations, we propose that angiotensin II affects the adrenal glomerulosa cell by first interacting with receptors, then increasing phosphatidyl inositol turnover, then altering cellular calcium distribution. Step distal to altered calcium distribution that contribute to increased steroid output include altered phospholipid metabolism, protein synthesis, and Na/k metabolism. PMID- 6279509 TI - Immunohistochemical evidence that angiotensins I and II are formed by intracellular mechanism in juxtaglomerular cells. AB - The existence of angiotensin II (AII) immunoreactivity in juxtaglomerular (JG) cells of rat kidney, which has been demonstrated previously by immunohistochemical studies, can be explained either as the product of intracellular synthesis or by the internalization of receptor-bound AII originating in plasma. To resolve these two alternative mechanisms, attempts were made to identify AI in JG cells of rat kidney by immunohistochemical staining using specific antibodies to AI. Although AI-like immunoreactivity was not detected in normal rat kidney, rats treated with the angiotensin-converting enzyme inhibitors, MK-421 or captopril, showed AI-like immunoreactivity in JG cells. The presence of renin and AII-like immunoreactivity was demonstrated in the same cells by specific antibodies to respective antigens used on adjacent serial sections. These findings support an intracellular mechanism of the formation of AII and suggest an intracellular renin angiotensin system, presumably separate from the extracellular system. PMID- 6279510 TI - Mutant of Bordetella pertussis which lacks ability to produce filamentous hemagglutinin. AB - Mutant cultures that lacked the ability to produce filamentous hemagglutinin were obtained by UV irradiation of Bordetella pertussis strain 18-323. Various biological activities of these mutants were compared with those of the parent culture. It was found that the leukocytosis-promoting, histamine-sensitizing, and mouse-protecting activities of the mutants were similar to those of the parent strain but that the virulence to mice was lower. The results seemed to rule out fimbrial hemagglutinin as a protective antigen when the ability to protect mice was measured by the intracerebral challenge test. PMID- 6279511 TI - Effect of infectious bovine rhinotracheitis virus infection on bovine alveolar macrophage function. AB - Bovine alveolar macrophages isolated in culture were assessed for immunological activity in assays for Fc and complement receptors, for phagocytosis, and for effector cell function in antibody-dependent cell cytotoxicity. In the case of uninfected alveolar macrophages, Fc receptors were detected on approximately 94% of macrophages and complement receptors were detected on 39%. Phagocytosis of immunoglobulin G-coated sheep erythrocytes occurred in 58% of macrophages, and phagocytosis of opsonized Candida parapsilosis, mediated by the complement receptor, was observed in 68% of cells. Alveolar macrophages were efficient effector cells in antibody-dependent cell cytotoxicity. Infection of macrophages with infectious bovine rhinotracheitis (IBR) virus resulted in reductions in Fc mediated receptor activity and phagocytosis after approximately 12 and 6 h, respectively. Complement receptor activity was initially elevated and then markedly reduced. Macrophages retrieved from IBR-immune and -susceptible donors were affected to a similar extent. The ability of macrophages to participate in antibody-dependent cell cytotoxicity was reduced dramatically from 2 h after IBR virus infection, suggesting that IBR virus-infected alveolar macrophages undergo alterations in immunological activity long before morphological changes in the cells become apparent. PMID- 6279512 TI - Neurological involvement in mice after infection with a cold-adapted herpes simplex type 2 virus. AB - Experimental intracerebral infection of 4-week-old mice with the MS strain of herpes simplex virus type 2 or its derivative cold variant was compared. The infectious process was followed in both the brain tissue and the trigeminal ganglia, using hematoxylin and eosin and antigenic tracing with indirect peroxidase-antiperoxidase staining. The wild-type virus elicited a severe meningitis and necrotic lesions by 7 days post-inoculation in the brain. The cold variant produced a mild meningitis and no necrotic lesions. In both viral infections, the neuron seemed to be the target cell at the early stages. Infection with the wild-type strain was able to spread through host tissues and produce confluent lesions, whereas infection with the cold mutant seemed to be confined to individual neurons. Data suggest that the cold variant is restricted in its ability to lyse the cells and spread in the host nervous tissue. PMID- 6279513 TI - Initial herpes simplex virus type 1 infection prevents ganglionic superinfection by other strains. AB - The ganglia of rabbits infected with a relatively benign strain of herpesvirus (E 43) and challenged with either of two virulent neurotrophic strains (MP or McKrae) were found to be colonized only by the initial benign infecting strain. Primary infection with the E-43 strain resulted in milder disease when the animals were infected with MP or McKrae strains and also prevented colonization of the ganglion by these strains. Neutralization with anti-glycoprotein C, plaque morphology, cytopathic effects, reconstruction experiments, and restriction endonuclease analysis indicated that the virus recovered from the ganglion was the initial infecting E-43 strain; no traces of the challenging MP and McKrae strains were found. The challenging McKrae strain was shed for several weeks in a few animals, but the virus isolated from the trigeminal ganglia of these animals was the primary infecting E-43 strain. These results suggest that initial infection with a relatively benign strain of herpesvirus may prevent superinfection of the ganglion (but not necessarily the end organ) by highly virulent herpes simplex virus strains and could have significant implications in the consideration of immunization against this disease in humans. PMID- 6279514 TI - Uncoupled relationship between demyelination and primary infection of myelinating cells in Theiler's virus encephalomyelitis. AB - Theiler's virus infection in mice produces a chronic demyelinating disease which appears to be based on an immune pathogenesis rather than on direct viral destruction of myelin-supporting cells. The purpose of the present study is to ascertain whether viral antigen is present in the cytoplasm of such cells in areas of demyelination. Because of the difficulty of identifying oligodendrocytes in tissues rich in infiltrating mononuclear cells and fixed for immunohistochemistry, I turned to a recently described form of Theiler's virus encephalomyelitis which follows inoculation with the attenuated ww strain and is characterized by extensive spinal cord remyelination by invading Schwann cells and by recurrent demyelination of Schwann cell-remyelinated axons. The unlabeled antibody peroxidase-antiperoxidase technique was employed to study whether such spinal cord Schwann cells were primarily infected by virus at the time when recurrent demyelination was occurring. Whereas other types of cells, including neurons, astrocytes, and macrophages, contained abundant viral antigen, no positive immune reaction was observed in Schwann cells. These results correlate with our previous studies which had suggested that demyelination in this viral model is not dependent on primary viral attack on myelinating cells but is probably dependent on the host immune response. PMID- 6279515 TI - Collagenolytic activity of Vibrio vulnificus: potential contribution to its invasiveness. AB - Vibrio vulnificus (lactose-positive vibrio) produced collagenase when grown in 2% synthetic sea salts supplemented with hydrolyzed casein. The addition of collagen or peptone to the medium increased the level of collagenase production. Collagenase activity was inhibited by EDTA but not by fetal calf serum. PMID- 6279516 TI - Cell-mediated immunity in human cytomegalovirus infection. AB - The direct leukocyte migration inhibition test, in response to cytomegalovirus stimulation, was used to study cell-mediated immunity in a group of children with cytomegalovirus infection. The test was impaired in children with chronic disease associated with cytomegaloviruria. In those cases with no viruria at the moment of the test, leukocyte migration inhibition was normal. Our data suggest that the acquired chronic cytomegalovirus infection may be sustained by a state of specific cellular desensitization, as already demonstrated for congenital infection. PMID- 6279517 TI - Isolation and characterization of isogenic pairs of domed hemolytic and flat nonhemolytic colony types of Bordetella pertussis. AB - Four different serotype strains of Bordetella pertussis, 3779BL(2)S(4), Tohama I, 353/Z, and 2753, were plated on Bordet-Gengou agar, where they grew as domed, hemolytic (D(+)H(+)) wild-type colonies. Cloned D(+)H(+) colony types of all four strains were passed onto modified Stainer-Scholte medium solidified with 1% Noble Agar. Colonies were selected from Stainer-Scholte agar, and these subsequently grew as flat, nonhemolytic (D(-)H(-)) colonies when transferred back onto Bordet Gengou agar. The frequency of D(-)H(-) organisms within a population of cloned D(+)H(+) was determined to be between 5 x 10(-5) and 5 x 10(-6). The D(-)H(-) colony types maintained their flat, nonhemolytic characteristics for over 80 single-colony passages on Bordet-Gengou agar. The isogenic pairs of D(+)H(+) and D(-)H(-) colony types from the four strains were compared for hemagglutination titer, lymphocytosis-promoting activity, adenylate cyclase activity, and presence of agglutinogens by agglutination. In all cases the D(-)H(-) colony types showed reduced activities or amounts of antigen compared with their D(+)H(+) parents. Freely diffusible antigens were markedly different between the two phenotypes as noted by double diffusion of antisera added to plates on which colonies of the variants were growing. Antigens solubilized from the two colony types by Triton X 100 were also markedly different as judged by radial immunodiffusion with antifimbrial hemagglutinin, antilymphocytosis-promoting factor, and anti-353/Z adsorbed with autoclaved 353/Z. In addition, autoradiographs of (125)I-surface labeled whole cells separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed unique banding patterns for each colony type. Since all organisms, regardless of colony type, were grown on Bordet-Gengou agar, the differences reported could not be due to medium composition. Differences between phenotypes were also independent of passage number on Bordet-Gengou agar. By analogy to previous studies, the D(-)H(-) organisms appear to fulfill the criteria for phase III or phase IV in the system of Leslie and Gardner (P. H. Leslie and A. D. Gardner, J. Hyg. 31:423-434, 1931) or phase III in the system of Kasuga et al. (T. Kasuga, Y. Nakase, K. Ukishima, and K. Takatsu, Kitasato Arch. Exp. Med. 26:121-134, 1954). PMID- 6279518 TI - Involvement of a plasmid in the invasive ability of Shigella flexneri. AB - Representative Shigella flexneri strains were studied to determine whether plasmids are involved in their virulence. All invasive S. flexneri strains, irrespective of serotype, were found to harbor a large plasmid of approximately 140 megadaltons in size, although some strains carried additional plasmid species. Spontaneous variants of strains of serotypes 1, 2, and 5 had lost this 140-megadalton plasmid and had concomitantly become avirulent, i.e., could neither invade HeLa cell monolayers nor produced keratoconjunctivitis in guinea pigs. To monitor plasmid transfer, the 140-megadalton plasmid of strain M90T (serotype 5) was tagged with the kanamycin resistance transposon Tn5. This tagged plasmid, pWR110, was not self-transmissible, but was mobilized by one of several different conjugative plasmids into avirulent derivatives of the heterologous serotypes 1 and 2 which had lost the comparable large plasmid. Transconjugants of both serotypes which had received pWR110 regained virulence. These data directly demonstrate that this large S. flexneri plasmid encodes or regulates some function(s) required for epithelial cell penetration. PMID- 6279520 TI - Mecillinam versus cephaloridine for the treatment of acute pyelonephritis. PMID- 6279519 TI - Production of mononuclear cell chemotactic factors during Sindbis virus infection of mice. AB - Draining lymph node cells of mice infected subcutaneously with Sindbis virus (SV) produced two mononuclear chemotactic factors in vitro. One factor did not require the addition of SV in vitro and was only detectable during the first week after infection. A second factor, resembling lymphocyte-derived chemotactic factor, required the addition of SV in vitro, was first detectable at 3 days, reached a peak at 15 to 18 days, and was gone by 29 days after infection. The production of this factor was virus specific. Diluent-inoculated mice produced no detectable mononuclear chemotactic factors in response to SV. In vitro production of the virus-specific chemotactic factor was dependent on both adherent cells and sensitized Lyt1+ T cells. In vitro production of the spontaneous factor was associated only with adherent cells but also appeared to be T cell dependent, since the lymph node cells from SV-infected athymic nude mice failed to produce either factor. Infectious center assays showed that adherent cells contained infectious SV without replicating it, suggesting the engulfment of virus by macrophages in the lymph node draining the area of virus replication. These cells probably process virus as antigen for presentation to T cells, resulting in local production of chemotactic factors as well as production in more distant sites of viral replication after leaving the lymph node. These virus-stimulated, mononuclear cell-produced chemotactic factors are likely to be of importance in generating the mononuclear inflammatory response. PMID- 6279521 TI - Pivmecillinam plus pivampicillin in urinary tract infections. A double-blind comparison with pivmecillinam alone in hospitalized patients. AB - Forty hospitalized patients with mostly uncomplicated urinary tract infections received treatment for ten days with either pivmecillinam/pivampicillin of pivmecillinam alone. The bacteriological cure rate immediately after therapy was discontinued was 82% (18/22) in the combination group and 72% (13/18) in the pivmecillinam group. The causative organisms were Enterobacteriaceae in 89% of the cases. All but one were sensitive in vitro to mecillinam alone. No side effects were recorded. PMID- 6279522 TI - [Eosinophilic meningomyelitis (author's transl)]. AB - Eosinophilia of more than 50% was observed for a period of ten days in the cerebrospinal fluid (CSF) of a patient with acute meningomyelitis and paraplegia. Serologic studies suggested the simultaneous infection with mycoplasma pneumoniae and Coxsackie A 09 as being a likely cause of meningomyelitis. This case demonstrates that pronounced CSF eosinophilia may be observed for a short period in non-parasitic infections of the central nervous system. parasitic infestation may be assumed only when eosinophilia persists for several months. PMID- 6279523 TI - Antibacterial activity of N-formimidoyl thienamycin in comparison with cefotaxime, lamoxactam, cefoperazone, piperacillin gentamicin. AB - The in vitro activity of N-formimidoyl thienamycin (MK 0787), a new stable derivative of thienamycin was compared with that of lamoxactam, cefotaxime, cefoperazone, piperacillin and gentamicin against 410 clinical isolates of common bacteria. In comparison with the other agents, MK 0787 was more or equally active against Staphylococcus aureus, Streptococcus faecalis, Escherichia coli, Citrobacter spp., Klebsiella spp., and Serratia spp. It inhibited all isolates at a concentration of 0.5 mg/l, but was less active than cefotaxime and lamoxactam against Proteus mirabilis and less active than cefotaxime, lamoxactam and cefoperazone against Beta-lactamase positive Haemophilus influenzae. Strains of Pseudomonas aeruginosa were more susceptible to N-formimidoyl thienamycin than to other antibiotics. PMID- 6279524 TI - Perspectives in the pathogenesis and management of acute renal failure. PMID- 6279525 TI - 2' and 3' ribonucleoside monophosphate in leukocytes of acute myeloid leukemia: markers for early diagnosis of relapse. AB - Levels of 2' and 3' purine and pyrimidine ribonucleoside monophosphates (2'-, 3' NMP) in leukocytes from blood and/or bone marrow were measured in three adult patients with acute non-lymphoblastic leukemias. The measurements of 2'-, 3'-NMP were made by high-performance liquid chromatography (HPLC) at various times in the course of the disease. Complete remission (CR) was obtained for all three patients but two of these have since died after relapsing at 8 and 9 months, respectively. The third patient remains in CR at 1 1/2 year. The levels of 2'-, 3'-NMP in the leukocytes of the patient remaining in remission have not changed since the beginning of his remission. However, in the patients who relapsed 2'- and 3'-NMP levels increased first in bone marrow than in blood leukocytes. These increases occurred about 3 months before the relapse was detected by morphological criteria. These data suggest that 2'-, 3'-NMP measurements may have a prognostic value if used to monitor patients with acute myeloid leukemia in CR. PMID- 6279526 TI - Complement-fixing antibody to Epstein-Barr virus soluble antigen in populations at high and low risk for nasopharyngeal carcinoma. AB - Greenland Eskimos comprise an ethnic group with one of the highest recorded incidence rates for nasopharyngeal carcinoma in the world. Sera from 625 Eskimos and 73 Danes (Caucasians) living in Greenland, as well as from 62 Danes living in Denmark, were tested for complement-fixing antibody to Epstein-Barr virus (EBV) soluble antigen and, from this study group, 129 donors were matched by age and sex for a study comparing antibody to viral capsid antigen, early antigen, and soluble antigen. Both Eskimos and Danes living in Greenland had significantly higher titers of EBV antibodies than Danes living in Denmark, suggesting that environment was more important than genetics or socio-economic factors in determining the antibody response to EBV. Age and sex were also factors, higher titers occurring in females and young Eskimos. PMID- 6279527 TI - Detection of simian virus 40 surface-associated large tumor antigen by enzyme catalyzed radioiodination. AB - To facilitate detection of SV40 surface-associated tumor antigen (T-ag), conditions were established to surface label T-ag on intact cells by lactoperoxidase-catalyzed radioiodination (125I/LPO). SDS-PAGE analysis of anti-T immunoprecipitates of SV40-transformed and -infected cells labelled with 125I/LPO revealed the presence of iodinated T-ag. Several types of control experiments were employed to guarantee the surface specificity of the 125I/LPO labelling technique. When SV40-transformed mouse cells were surface labelled with lactoperoxidase and glucose oxidase immobilized on insoluble beads, a preparation less readily internalized than soluble enzymes, T-ag was iodinated. Selective immunoprecipitation of surface antigens demonstrated that lactoperoxidase did not iodinate internally localized T-ag. A reconstruction experiment in which an extract of SV40-infected cells was added to uninfected cells prior to surface labelling suggested that T-ag released from lysed cells did not adhere significantly to monolayer surfaces and become iodinated. Finally, systematic omission of reactants from the iodination reaction revealed that exogenous addition of lactoperoxidase and H2O2 was necessary to generate an iodinated T-ag, indicating that endogenous host cell reactants do not contribute significantly to the iodination of T-ag. 125I-labelled T-ag was detectable on the surface of SV40 tsA-infected cells at the nonpermissive temperature 24 h post infection, indicating that the tsA lesion does not prevent the interaction of T-ag with the cell surface. When 125I/LPO-labelled transformed or infected cells were chased for 2.5 h after labelling, iodinated T-ag was no longer associated with the cell monolayer but was immunoprecipitable from culture supernatants. Cultures from which labelled T-ag had been shed could then be relabelled with 125I/LPO and surface-associated T-ag was again detectable. These data suggest that surface associated T-ag is continuously shed from the cell surface and is rapidly replaced in the membrane by intracellular T-ag. PMID- 6279528 TI - Characterization of a highly oncogenic murine leukemia virus from wild mice. AB - We have isolated a highly lymphomagenic wild mouse virus by passage of a weakly oncogenic amphotropic murine leukaemia virus (MuLV-A) in NIH Swiss mice. This virus is similar in host range, interference and neutralization properties to that of the inoculated amphotropic virus but is distinct from it biochemically and causes lymphomas in 90-100% of mice within 1-2 months. Our results indicate that this highly oncogenic virus is a recombinant of wild mouse MuLV-A and sequences related to env gene region of the endogenous xenotropic virus of NIH Swiss mice. PMID- 6279529 TI - Lithium and warts. PMID- 6279530 TI - Cell-mediated immunity in wart infection. AB - The role of cell-mediated immunity (CMI) in wart infection was demonstrated. The correlation with humoral immunity was also suggested. Patients with cell-mediated immune deficiency were found to be more susceptible to wart infection than those with humoral immune deficiency. PMID- 6279531 TI - Changes in water content and volume accompanying exsheathment of Haemonchus contortus. PMID- 6279532 TI - Hydroxyl radical damage in low oxygen concentrations in irradiated bacteria. PMID- 6279533 TI - E.S.R. of spin-trapped radicals in aqueous solutions of pyrimidine nucleosides and nucleotides. Reactions of the hydroxyl radical. AB - Free radicals produced by the reactions of OH radicals with ribose phosphates, pyrimidine nucleosides and nucleotides in aqueous solutions have been investigated by e.s.r. and spin-trapping techniques. OH radicals were generated by U.V. -photolysis of hydrogen peroxide and short-lived free radicals of the samples were spin-trapped by t-nitrosobutane and identified by e.s.r. spectroscopy. For ribose-5'-phosphate and 2' -deoxyribose-5' -phosphate, e.s.r. spectra observed consisted mainly of singles due to -C(5')H2 radicals which were initiated by H-abstraction from the sugar at the C-4' position and formed by the radical transfer to the C-5' position. It has also been shown that OH radicals abstract a hydrogen atom from the sugar at C-1' and C-5' positions. For cytidine, deoxycytidine, 5'-CMP, 3' -CMP and 5' -dCMP, four radicals have been observed and for uridine, deoxyuridine, 5' -UMP, 3' -UMP and 5' -dUMP, the existence of at least three radical species have been established. In contrast to the case of ribose phosphates, no signals due to -C(5')H2 radicals were detected for pyrimidine nucleosides and nucleotides. The results are discussed in relation to a recent mechanism which described OH-induced strand breaks of DNA. PMID- 6279534 TI - Influence of anoxic sensitizers on the radiation damage in biologically active DNA in aqueous solution. AB - The competition between biologically active single-stranded phiX174 DNA and the anoxic radiosensitizers metronidazole, misonidazole, paranitroacetophenone or nifuroxime for OH radicals is studied. The results are compared with experiments in which the protection of the DNA by t-butanol is determined. Also the effects of the sensitizers on the chemical nature of the damage (immediate and potential break, immediate and potential base damage) is studied. It is found that in diluted aqueous solutions of DNA these radiosensitizers do not sensitize with respect to the biological inactivation. The only effect observed is a shift from potential to immediate breaks with misonidazole and also nifuroxime. PMID- 6279535 TI - Recent trends in surgery of peripheral nerves. PMID- 6279536 TI - Evaluation and prediction of regenerative processes by quantitative analysis of the electromyographic response and biofeedback in peripheral nerve injuries. AB - The purpose of this study was to quantitatively and objectively evaluate and monitor regenerative processes in peripheral nerve lesions using a new electrophysiological method, and to develop and evaluate an electromyographic (EMG) biofeedback training programme for the rehabilitation of patients with such injuries. Five patients with traumatic unilateral brachial plexus injuries participated in the study longitudinally on a twice-weekly basis. The electrophysiological method was based on quantitative on-line analyses of the myoelectric signal from affected muscles of the upper limb. The biofeedback programme was developed and used to: (1) increase maximum voluntary centrally mediated neuromuscular activity; (2) dissociate desired and interfering neuromuscular activity; and (3) improve patient control over the neuromuscular activity. The electrophysiological results have differentiated clearly between muscles where regeneration can be assumed to be in progress and muscles where it is not in progress. This differentiation can be made prior to the occurrence of any observable contraction in muscle under study. The biofeedback training resulted in improved neuromuscular function in all patients. PMID- 6279537 TI - Plasma cyclic AMP hyper-response to glucagon in manic patients on lithium treatment. AB - We have measured the rise in plasma cyclic AMP induced by glucagon and adrenaline in manic patients on lithium therapy. Glucagon-induced increases in the concentration of plasma cyclic AMP were larger, but the adrenaline-induced increases were smaller, in lithium-treated manic patients than in normal controls. In 3 manic patients, the enhanced response to glucagon was normalized 3 weeks after the cessation of lithium treatment. Though the underlying mechanism is unknown, the unusual plasma cyclic AMP responses may reflect altered receptor sensitivity which would be closely related to the curative effect of lithium on symptoms of mania. PMID- 6279538 TI - [Roentgen diagnosis of the pancreas]. PMID- 6279540 TI - Role of the cortical neuron: integrator or coincidence detector? AB - A model was constructed of the relations between the incoming excitatory postsynaptic potentials and inhibitory postsynaptic potentials to the intracellular membrane potential fluctuations and to the firing rate of a single neuron. From this model, the strength of synapses was assessed in two ways: 1) the ability of several synchronous presynaptic spikes to initiate a postsynaptic spike, termed the synchronous gain of the synapse; and 2) the ability of several asynchronous presynaptic spikes to add a spike to the output spike train, called the asynchronous gain. It was found that for the conditions prevailing in the brain's cortex, the synchronous gain is almost always higher. It appears that the cortical neurons act as coincidence detectors, and that the appropriate code for the higher cortical functions is coincidence and not firing rate. PMID- 6279539 TI - Inhibition of sensory transmission in the cat lemniscal system during fast movements: a review. PMID- 6279541 TI - Comparison between biochemical and histochemical assessments of peroxidase activity in rat mammary tumours. AB - The endogenous peroxidase content of 26 'hormone-dependent' and 16 'hormone independent' rat mammary tumours was assessed by means of a biochemical (guaiacol) assay on tumour extracts and by means of a histochemical (diaminobenzidine) technique on frozen sections of the same tumours. By guaiacol assay, the hormone-dependent tumours had significantly higher peroxidase levels than the hormone-independent tumours. In contrast, by diaminobenzidine staining of the same tumours, peroxidase was detectable in 94% of hormone-independent tumours but in only 54% of hormone-dependent tumours. Moreover, there was no direct correlation between the results of biochemical and histochemical methods. At least in these rat mammary tumours, therefore, histochemical estimates of peroxidase activity based on the diaminobenzidine reaction do not seem to reflect the same tissue properties as biochemical estimates based on the guaiacol reaction after tissue disruption. PMID- 6279542 TI - Cytochemical markers of bladder carcinogenesis. AB - Enzyme cytochemical and immunocytochemical techniques at the light and electron microscope levels were used to study the distribution of potential markers of chemical transformation in rodent bladders. In rat tumours induced by in vivo treatment with methylnitrosourea, alkaline phosphatase localization was normal on the external surface of the plasma membranes of some cells but abnormal in others where reaction product was seen only on intracellular membranes. 5'-Nucleotidase localization was abnormal in all cells, being seen on endoplasmic reticulum and nuclear membranes only, while in normal bladders only ectoenzyme localization was seen. Heterogeneity of alkaline phosphatase amd 5'-nucleotidase localization was seen on the plasma membranes of these tumours after 15 days in organ culture. Some cells produced enzyme and others did not; in other cells only parts of the membrane reacted heavily, while other regions were negative. In transformed cell cultures and tumours of mouse bladder derived by in vitro treatment of explants with dimethylbenz (a) anthracene, a bimodal pattern of alkaline phosphatase localization was seen. Cells had either normal ectoenzyme reaction product or abnormal intracellular membrane reaction product. 5'-Nucleotidase and ADPase were lost after transformation while cAMP-phosphodiesterase was retained as an ectoenzyme. Mg.ATPase and a cAMP-independent, calcium-insensitive 'protein phosphatase' were induced in transformed cell cultures. An epithelial antigen was detected in the cytoplasm of both normal and transformed cells associated with reticular cytoplasmic ground substance, plasma membrane vesicles and cytoskeletal elements. PMID- 6279544 TI - Studies on DNA content of hepatocytes in cirrhosis and hepatoma by means of microspectrophotometry and radioautography. AB - In order to clarify the underlying mechanism of malignant transformation from cirrhosis to hepatoma the cell kinetics of hepatocytes were studied in these two conditions. The content and synthesis of DNA in hepatocyte nuclei were investigated, by means of Feulgen-microspectrophotometry and tritiated thymidine radioautography, in cirrhotic and noncancerous parts of hepatoma with concomitant cirrhosis. The distribution of ploidy patterns was widely spread, from hypodiploid to hyperpolyploid, in the noncancerous parts of a cirrhotic liver containing hepatoma. In normal liver, each paired nuclear DNA content of a binucleate cell recorded almost the same amount, whereas in the noncancerous as well as in hepatoma cells much difference of DNA content was observed between the paired nuclei of the binucleate cells. The ploidy pattern of hepatocytes in patients with liver cirrhosis, who had developed hepatoma during follow-up periods of several months to several years, appeared to resemble that in noncancerous parts of hepatoma cases. On the other hand, the incorporation of tritiated thymidine into hepatocytes was found to be markedly increased in noncancerous parts as well as in cirrhotic liver developing hepatoma during follow-up periods. These results suggest the possibility that the hepatocytes in noncancerous parts of hepatoma have deranged cell-kinetics which might be a driving factor for the development of malignancy. PMID- 6279543 TI - Immunocytochemical localization of Na+, K+-ATPase in the rat kidney. AB - To determine if rat kidney Na+, K+-ATPase can be localized by immunoperoxidase staining after fixation and embedding, we prepared rabbit antiserum to purified lamb kidney medulla Na+, K+-ATPase. When sodium dodecylsulfate polyacrylamide electrophoretic gels of purified lamb kidney Na+, K+-ATPase and rat kidney microsomes were treated with antiserum (1:200), followed by [125I]-Protein A and autoradiography, the rat kidney microsomes showed a prominent radioactive band coincident with the alpha-subunit of the purified lamb kidney enzyme and a fainter radioactive band which corresponded to the beta-subunit. When the Na+, K+ ATPase antiserum was used for immunoperoxidase staining of paraffin and plastic sections of rat kidney fixed with Bouin's, glutaraldehyde, or paraformaldehyde, intense immunoreactive staining was present in the distal convoluted tubules, subcapsular collecting tubules, thick ascending limb of the loops of Henle, and papillary collecting ducts. Proximal convoluted tubules stained faintly, and the thin portions of the loops of Henle, straight descending portions of proximal tubules, and outer medullary collecting ducts did not stain. Staining was confined to basolateral surfaces of tubular epithelial cells. No staining was obtained with preimmune serum or primary antiserum absorbed with purified lamb kidney Na+, K+-ATPase, or with osmium tetroxide postfixation. We conclude that the basolateral membranes of the distal convoluted tubules and ascending thick limb of the loops of Henle are the major sites of immunoreactive Na+, K+-ATPase concentration in the rat kidney. PMID- 6279545 TI - Biochemical and histopathological response to chrysotile ingestion in guinea pigs. PMID- 6279546 TI - Respiratory and inert gas exchange during high-frequency ventilation. AB - Pulmonary gas exchange during high-frequency low-tidal volume ventilation (HFV) (10 Hz, 4.8 ml/kg) was compared with conventional ventilation (CV) and an identical inspired fresh gas flow in pentobarbital-anesthetized dogs. Comparing respiratory and infused inert gas exchange (Wagner et al., J. Appl. Physiol. 36: 585--599, 1974) during HFV and CV, the efficiency of oxygenation was not different, but the Bohr physiological dead space ratio was greater on HFV (61.5 +/- 2.2% vs. 50.6 +/- 1.4%). However, the elimination of the most soluble inert gas (acetone) was markedly enhanced by HFV. The increased elimination of the soluble infused inert gases during HFV compared with CV may be related to the extensive intraregional gas mixing that allows the conducting airways to serve as a capacitance for the soluble inert gases. Comparing as exchange during HFV with three different density carrier gases (He, N2, and Ar), the efficiency of elimination of Co2 or the intravenously infused inert gases was greatest with He O2. However, the alveolar-arterial partial pressure difference for O2 on He-O2 exceeded that on N2-O2 by 5.4 Torr during HFV. The finding agrees with similar observations during CV, suggesting that this aspect of gas exchange is not substantially altered by HFV. PMID- 6279547 TI - A multidimensional approach to evaluation. PMID- 6279550 TI - Organizing approaches used in curriculum design. PMID- 6279549 TI - Role transition: a practicum for baccalaureate nursing students. PMID- 6279551 TI - Process evaluation: a second look at psychomotor skills. PMID- 6279548 TI - Interdisciplinarism: problems & promises. PMID- 6279552 TI - Nursing continuing education: California to Texas via teleconference. PMID- 6279553 TI - Border disease in a flock of Minnesota sheep. AB - A flock problem affecting newborn lambs on a Minnesota farm was investigated. The lambs had hairy fleece and rhythmic tremors, and were unthrifty. Histologic examination revealed hypomyelinization of the CNS and hypertrophy of primary follicles in the skin. A virus antigenically related to bovine viral diarrhea virus was isolated. Serum neutralization showed that a large number of flock ewes had much higher titers to bovine viral diarrhea virus than did control animals. PMID- 6279554 TI - Poly(ADP-ribose) polymerase activity in HeLa cells treated with various antibiotics. PMID- 6279555 TI - Examination of bovine faeces for the isolation and identification of Clostridium species. PMID- 6279556 TI - Resistance to chloramphenicol and metronidazole in anaerobic bacteria. PMID- 6279557 TI - Cholecalciferol--a vitamin or hormone. PMID- 6279558 TI - Transformation of Rhizobium meliloti 41 with plasmid DNA. AB - Plasmid pGV1106, a derivative of the wide-host-range plasmid S-a of the W incompatibility group, was introduced into Rhizobium meliloti 41 by plasmid mediated mobilization to overcome the restriction of foreign DNA. The mobilized plasmid pKK2 differed from the original pGV1106 by an extra piece of DNA of 1.3 kilobase pairs which supposedly originated from pJB3JI used for mobilization. If pKK2 was isolated from R. meliloti 41, it could be successfully reintroduced by transformation. The transformation frequency was low (10 to 54 colonies per micrograms of plasmid DNA) but reproducible, and several lines of evidence showed that it was the consequence of plasmid DNA uptake. The small size (10.3 kilobases) and elevated copy number (10 to 15 copies per cell) of pKK2 make it a potentially useful cloning vector for the study of symbiotic nitrogen fixation genes of R. meliloti 41. PMID- 6279559 TI - Transposition of the carbenicillin-hydrolyzing beta-lactamase gene. AB - We isolated a new transposon Tn2101, from plasmid Rms433 in Enterobacter cloacae. Tn2101 encoded the formation of type IV (carbenicillin-hydrolyzing) beta lactamase and multiple resistance to streptomycin, sulfanilamide, spectinomycin, and mercury in addition to ampicillin. Tn2101 was transposable between conjugative (or nonconjugative) plasmids and the host chromosome. Transposition occurred independently of the general recombination ability of the host cell. Tn2101 had a molecular size of 9.5 x 10(6) and contained short inverted repeat terminal sequences. PMID- 6279560 TI - Second system for potassium transport in Streptococcus faecalis. AB - It has been reported that the accumulation of K+ by Streptococcus faecalis is mediated by a transport system which required both ATP and the proton motive force (Bakker and Harold, J. Biol. Chem. 255:433-440, 1980). My results indicate that S. faecalis has a second transport system for K+. The features of this system are as follows: (i) the system is driven by ATP (or a derivative of ATP) and does not require the proton motive force; (ii) the system is normally absent in the wild-type strain but can be derepressed by lowering rhe intracellular concentration of K+; (iii) the pH optimum of this system is about 8.5, and no detectable K+ is accumulated at pH values below 6.5; and (iv) the rate of Rb+ accumulation by this system is very low. These properties are quite different from those of the transport system described by Bakker and Harold. Therefore, I propose that S. faecalis has two K+ transport systems. PMID- 6279561 TI - Properties of R1162, a broad-host-range, high-copy-number plasmid. AB - Regions of plasmid DNA encoding characteristic properties of the IncQ (P-4) group plasmid R1162 were identified by mutagenesis and in vitro cloning. Coding sequences sufficient for expression of incompatibility and efficient conjugal mobilization by plasmid R751 were found to be linked to the origin of DNA replication. In contrast, there was a region remote from the origin, and active in trans, that was required for plasmid maintenance. A derivative that was temperature sensitive for stability was isolated. The defect mapped at or near the region required for plasmid maintenance and resulted in far fewer copies of supercoiled plasmid DNA per cell under permissive conditions. A second region required for stability was also identified from the behavior of a deletion derivative of R1162, which did not, however, show an altered number of supercoiled plasmid DNA copies. Finally, a plasmid DNA mutation resulting in a substantially higher copy number was isolated. Plasmid reconstruction experiments suggested that the mutation was linked to the replicative origin. PMID- 6279562 TI - Proton motive force and the physiological basis of delta pH maintenance in thiobacillus acidophilus. AB - At optimal growth pH (3.0) Thiobacillus acidophilus maintained an internal pH of 5.6 (delta pH of 2.6 units) and a membrane potential (delta psi) of some +73 mV, corresponding to a proton motive force (delta p) of -83 mV. The internal pH remained poised at this value through external pH values of 1 to 5, so that the delta pH increased with decreasing external pH. The positive delta psi increased linearly with delta pH: above a delta pH of 0.6 units, some 60% of the increase in delta pH was compensated for by an opposing increase in delta psi. The highest magnitude of delta pH occurred at an external pH of 1.0, where the cells could not respire. Inhibiting respiration by CN- or azide in cells at optimal pH decreased delta pH by only 0.4 to 0.5 units and caused a corresponding opposite increase in delta psi. Thus, a sizable delta pH could be maintained in the complete absence of respiration. Treatment of cells with thiocyanate to abolish the delta psi resulted in a time-dependent collapse of delta pH, which was augmented by protonophores. We postulate that T. acidophilus possesses unusual resistance to ionic movements. In the presence of a large delta pH (greater than 0.6 pH units), limited diffusion of H+ into the cell is permitted, which generates a positive delta psi because of resistance to compensatory ionic movements. This delta psi, by undergoing fluctuations, regulates the further entry of H+ into the cell in accordance with the metabolic state of the organism. The effect of protonophores was anomalous: the delta p was only partially collapsed, and respiration was strongly inhibited. Possible reasons for this are discussed. PMID- 6279563 TI - Phosphoenolpyruvate:sugar phosphotransferase system-mediated regulation of carbohydrate metabolism in Salmonella typhimurium. AB - The crr mutation was shown to affect the phosphoenolpyruvate:sugar phosphotransferase system-mediated transient repression of the lac operon, intracellular cAMP levels, and sensitivity to inducer exclusion. Our results indicate that the presumed crr gene product, factor IIIGlc, plays a direct role in the regulation of inducer exclusion. We propose a mechanism in which inducer exclusion depends on both the level and state of phosphorylation of factor IIIGlc and the level of an inducer exclusion-sensitive transport system. The results of studies on the sensitivity to inducer exclusion of glycerol and maltose in cultures induced for short periods of time on these substrates (resulting in varying degrees of activity of the respective transport systems) support this model of inducer exclusion. Previously, the crp*-771 mutation has been shown to result in an altered cAMP receptor protein, which has a changed affinity for cAMP, and to affect the sensitivity for inducer exclusion of glycerol. Changes in other functions of the altered cAMP receptor protein were indicated by our results; these changes were in the roles of this protein in (i) the cAMP dependent initiation of transcription of the lac operon and (ii) the regulation of intracellular cAMP levels and the export of cAMP. We propose that the crp*-771 mutation has an indirect effect in relieving inducer exclusion in repressed or hypersensitive strains, in which the crp*-771 mutation allows the synthesis of inducer exclusion-sensitive transport systems to higher levels than the levels found in strains containing wild-type cAMP receptor protein. PMID- 6279564 TI - Attachment of the chromosomal terminus of Bacillus subtilis to a fast-sedimenting particle. AB - After gently lysed protoplasts of exponential phase cells of Bacillus subtilis were treated with restriction endonuclease BamHI, 99% of the DNA did not sediment with the plasma membrane. This DNA was fractionated on sucrose gradients into (i) a fast-sedimenting fraction highly enriched for genes from the origin and terminus (purA and ilvA), (ii) a 50 to 100S component also enriched for purA and ilvA, and (iii) the bulk of the DNA. The fast-sedimenting fraction was dissociated by Sarkosyl; this fraction contained a substantial amount of protein and is probably a membrane subparticle. The S value of the 50 to 100S component was not greatly affected by Sarkosyl treatment, but these particles were unable to penetrate an agarose gel during electrophoresis and were retained by nitrocellulose filters. The terminus DNA in the fast-sedimenting fraction and the 50 to 100S component contained a large restriction fragment (1.5 x 10(7) to 2.0 x 10(7) daltons) encoding ilvA, thyB, and ilvD. The bulk of the SP beta prophage and metB, which lie to the right and left, respectively, of the ilvA-ilvD cluster, were not part of the complex. citK, which lies to the right of SP beta, appeared to be present in the fast-sedimenting complexes. The neighboring genes kauA and gltA were not part of the fast-sedimenting complexes. The presence of terminus DNA in the fast-sedimenting components was also demonstrated by a radiochemical method. PMID- 6279565 TI - A multifunctional gene (tetR) controls Tn10-encoded tetracycline resistance. AB - The tetracycline resistance regulatory gene (tetR) of transposon Tn10 was analyzed by a combination of methods involving gene fusion and cloning. This gene is located on a 695-base pair HincII DNA fragment near the center of Tn10. The direction of transcription is opposite to that of neighboring gene tetA, which encodes the TetA protein. The gene product of the tetR gene (the TetR protein) has a molecular weight of 23,000. tet-R-lacZ gene fusions encode fusion beta galactosidases that are membrane bound, indicating that the TetR protein itself is membrane associated. Mutants defective in tetR result in constitutive tetracycline resistance, but the level of resistance is reduced. Expression of the tetR gene is induced by tetracycline; in the absence of tetracycline, the TetR protein turns off its own synthesis. PMID- 6279567 TI - Identification of a new gene (secA) and gene product involved in the secretion of envelope proteins in Escherichia coli. AB - We have constructed lambda specialized transducing phages which carry an Escherichia coli gene (secA) involved in the secretion of certain envelope proteins. These phage have been used to show that secA is a new gene to the clockwise side of envA. The secA mutation previously described, secA5l(Ts) (D. B. Oliver and J. Beckwith, Cell 25:765-772, 1981), is recessive to the wild-type allele. We have also isolated Tn5 insertions in the gene carried on the transducing phage to further define the gene. These phage were used to infect UV irradiated cells to allow the identification of the secA gene product as a 92 kilodalton polypeptide and to show that transcription of secA is clockwise relative to the E. coli genetic map. PMID- 6279566 TI - Adenine nucleotide and lysine transport in Chlamydia psittaci. AB - Isolated reticulate bodies of Chlamydia psittaci were found to transport ATP and ADP by an ATP-ADP exchange mechanism. ATP uptake activity was not detected in elementary bodies. The apparent Km of transport for both ATP and ADP was approximately 5 microM, and the calculated Vmax for both was about 1 nmol of nucleotide transported per min per mg of protein. ADP competitively inhibited ATP transport with a Ki of 4.5 microM. Other nucleotides tested had no effect on the uptake of ATP. A magnesium-dependent, oligomycin-sensitive ATPase (ATP phosphohydrolase, EC 3.6.1.3) was associated with reticulate bodies, and most of the transported ATP was hydrolyzed to ADP, which was exchanged for additional, extracellular nucleotide. Some ADP was hydrolyzed to AMP, which exited the cells slowly. Lysine was transported against the electrochemical gradient by reticulate bodies in the presence of ATP. Oligomycin and carbonyl cyanide p trifluoromethoxyphenylhydrazone inhibited ATP-dependent lysine transport. Lysine exited reticulate bodies when the reticulate bodies were incubated in the presence of ADP, carbonyl cyanide p-trifluoromethoxyphenylhydrazone, or a reduced concentration of ATP. The results support the concept that chlamydiae are energy parasites which are capable of drawing upon the adenine nucleotides of their hosts, hydrolyzing ATP, and establishing an energized membrane. PMID- 6279568 TI - Facilitation of plasmid transfer in Streptococcus pneumoniae by chromosomal homology. AB - The frequency of plasmid establishment in the transformation of Streptococcus pneumoniae by plasmid DNA was increased more than 10-fold when the plasmid carried DNA homologous to the host chromosome. Perfect homology was not necessary for such facilitation; small additions or deletions were tolerated, but extensive deletions in the homologous segment of either plasmid or chromosome reduced or eliminated facilitation. The facilitated plasmid transfer showed a linear dependence on monomeric plasmid concentration rather than the quadratic dependence found in the absence of homology, which indicated that entering plasmid fragments interacted with the chromosome rather than with each other to establish a plasmid replicon. Restriction enzyme cleavage of the plasmid in the nonhomologous segment destroyed its activity, but cleavage in the homologous segment or even enzymatic removal of part of that segment did not prevent plasmid transfer, and plasmids of the original size were established. In facilitated transfer, chromosomal markers (additions and deletions as well as single-site mutations) entered the plasmid with a frequency ranging from 10 to 90% depending on the marker location. Several possible mechanisms for the establishment of plasmids in the presence of chromosomal homology and for the transfer of chromosomal information are considered. They depend on synapsis of the newly entered single-strand plasmid fragment with the host chromosome and subsequent copying of, donation from, or integration into the homologous chromosomal segment. After plasmid establishment, equilibration of donor and chromosomal markers between the chromosome and the plasmid pool, presumably by homologous recombination events, was observed. PMID- 6279570 TI - Trimethoprim-produced F-specific insertion mutations in Escherichia coli K-12. AB - Besides producing thymine-requiring mutants (thy), trimethoprim (TMP) cured the mini-ColE1 replicon pML21 at an appreciable frequency. The cured Escherichia coli K-12 cells behaved like polA mutants by failing to support the stable maintenance of the ColE1 plasmid. The mini-F replicon pSC138, which was lacking all three insertion sequences (IS3, gammadelta, and IS2) normally used for F-specific integration and excision, was not cured by TMP. Instead, it integrated into specific regions of the E. coli chromosome and thus caused auxotrophic mutations in operons which were always localized on either side of oriC (origin of chromosomal replication). The incompatibility and replication functions of the integrated plasmid in auxotrophs were retained, and the plasmid DNAs recovered from spontaneously occurring revertants did not show any alterations in their contour lengths as determined by electron microscopy. The F replicon (fragment 5) contained in plasmid pSC138 carried two origins of replication, the primary origin, oriV(1) at 42.6F and the secondary origin, oriV(2), at 44.1F. Another mini-F plasmid pMF21, deleted of the primary origin of replication (oriV(1)), was still capable of autonomous replication but failed to integrate onto the chromosome after TMP treatment. Furthermore, the composite plasmid pRS5, which normally uses only the replication origin and functions of the pSC101 component, was also insensitive to TMP. On the basis of these results, we propose a new scheme of F integration via the functional oriV(1) and suggest the involvement of a similar mechanism in the formation of Hfr strains by integrative suppression. PMID- 6279569 TI - Outer membrane protein P of Pseudomonas aeruginosa: regulation by phosphate deficiency and formation of small anion-specific channels in lipid bilayer membranes. AB - A new major outer membrane protein, P, was induced in Pseudomonas aeruginosa PAO1 upon growth in medium containing 0.2 mM or less inorganic phosphate. Studies with media containing different levels of phosphate and with mutants of PAO1 suggested that protein P was coregulated with alkaline phosphatase and phospholipase C. Protein P was substantially purified and shown to form sodium dodecyl sulfate resistant oligomers on polyacrylamide gels. The incorporation of purified protein P into artificial lipid bilayers resulted in an increase of the membrane conductance by many orders of magnitude. Single-channel experiments demonstrated that protein P channels were substantially smaller than all previously studied porins from P. aeruginosa and enteric bacteria, with an average single-channel conductance in 1 M NaCl of 0.25 nS. The protein P channel was apparently not voltage induced or regulated. The results of single-channel conductance experiments, using a variety of different salts, allowed a minimum channel diameter estimate of 0.7 nm. Furthermore, from these results it was concluded that the protein P channel was highly specific for anions. Zero-current potential measurements confirmed that protein P was at least 30-fold more permeable for Cl- than for K+ ions. The possible biological role of the small, anion-specific protein P channels in phosphate uptake from the medium is discussed. PMID- 6279572 TI - Analysis of Mycoplasma hyorhinis genome by use of restriction endonucleases and by electron microscopy. AB - The chromosome of Mycoplasma hyorhinis was analyzed by using different restriction endonucleases and electron microscopy. It was found that restriction enzymes BstEII, XhoI, and SacI are the enzymes of choice for analysis and characterization of M. hyorhinis. The bands resulting from digestion of M. hyorhinis DNA with BstEII had apparent molecular weights ranging from 1.2 X 10(6) to 75 X 10(6). The apparent total molecular weight of DNA was calculated from the molecular weights of the individual bands and found to be 251 X 10(6). Electron microscopic contour length measurements of the largest DNA fragments verified the molecular weight values calculated from gel analysis. Electron microscopic contour length measurements of intact DNA of M. hyorhinis revealed a molecular weight of 5.4 +/- 5 X 10(8). The discrepancy between the values of molecular weight of M. hyorhinis DNA as determined by restriction enzyme analysis and contour length measurement is based on the fact that some of the DNA fragments which migrate as an apparent single band in the agarose gel really are double or multiple DNA fragments. PMID- 6279571 TI - Evolution of the D-ribose operon on Escherichia coli B/r. AB - The D-ribose operon (rbs) of Escherichia coli K-12 maps at 83 min and is inducible. The rbs operon of E. coli B/r maps at 2 min and is constitutive. Evidence is presented showing that a second inducible copy of the rbs operons is present in E. coli B/r mapping at 83 min. The data indicated that the duplication of the rbs operon represented a transposition of the 83-min region to 2 min. The identification of a second copy of the rbs operon in B/r and the determination of its inducibility were based on the reactivation, through mutagenesis, of inducible rbs expression, mapping by P1 transduction of the mutation site to 83 min, and merodiploid complementation analysis of the D-ribokinase expression in E. coli B/r. We also show that the rbs transposition to 2-min continued to generate transposable elements coding for the 1- to 2-min region of the chromosome and transposing onto extrachromosomal DNA target molecules such as pBR322. PMID- 6279573 TI - Genetic characterization of the glutamate dehydrogenase gene (gdhA) of Salmonella typhimurium. AB - Salmonella typhimurium mutants, either devoid or glutamate dehydrogenase activity or having a thermolabile glutamate dehydrogenase protein, were used to identify the structural gene (gdhA) for this enzyme. Transductions showed that the mutations producing these phenotypes were linked to both the pncA and nit genes, placing the gdhA locus between 23 and 30 U on the S. typhimurium chromosome. Additional transductions with several Tn10 insertions established the gene order as pncA-gdhA-nit. Since few genetic markers exist in this region of the chromosome, Hfr strains were constructed to orient the pncA-gdhA-nit cluster with outside genes. Conjugation experiments provided evidence for the gene order pyrD pncA-gdhA-nit-trp. To further characterize gdhA, we used Mu cts d1 (Apr lac) insertions in this gene to select numerous strains containing deletions with various endpoints. Transductions of these deletions with strains containing different gdh mutations and with a mutant having a thermolabile glutamate dehydrogenase protein permitted us to construct a deletion map of the gdhA region. PMID- 6279575 TI - Changes in the organization of the outer membrane of Proteus mirabilis during swarming: freeze-fracture structure and membrane fluidity analysis. AB - Freeze-fracture studies of short, nonswarming Proteus mirabilis revealed the characteristic gram-negative profile of fractured inner membrane with densely packed particles and sectioned outer membrane with little or no fracture plane. Long swarming cells, however, fractured easily along both the inner membrane and a second membrane, probably the outer membrane. The inner membrane had a typical profile, whereas the outer membrane had fewer but more prominent particles. Isolation and purification of the inner and outer membranes of the short and long bacteria and examination of them with electron paramagnetic resonance measurements after spinlabeling supported the above observations. The outer membrane of swarmer cells allowed higher mobility of the spin label than did the outer membrane of the nonswarming short cells, which showed a typical rigid profile. These results suggest that regions of lipid bilayer appear in the outer membrane during swarmer formation. Previous observation of the behavior and biochemistry of P. mirabilis during swarming are discussed in light of these results. PMID- 6279574 TI - Nucleotide sequence and functional map of pE194, a plasmid that specifies inducible resistance to macrolide, lincosamide, and streptogramin type B antibodies. AB - pE194 is a small plasmid (isolated originally in Staphylococcus aureus) which confers erythromycin-inducible resistance to macrolide, lincosamide, and streptogramin type B (MLS) antibiotics. The nucleotide sequence of pE194 contains 3,728 base pairs (bp), corresponding to a molecular mass of 2.4 million daltons. By means of site-specific cleavage with restriction endonucleases and cloning resultant fragments, determinants of the two major biological functions of p E194, i.e., inducible MLS resistance and replication, could be localized and assigned to specific sequences in the plasmid. Restriction endonuclease TaqI cut pE194 at three sites. TaqI fragment A (1,443 bp) contained the determinant for inducible MLS resistance, whereas TaqI fragment B (1,354 bp) contained a determinant necessary for plasmid replication. Regulatory mutations resulting in constitutive expression of MLS resistance mapped in TaqI fragment A, whereas a mutation associated with elevated plasmid copy number was mapped in TaqI fragment B. Also mapping in TaqI fragment B was a plasmid replication determinant comprising two sets of inverted complementary repeat sequences, one of which spanned 124 bp and was adjacent to a second smaller set which was rich in guanine and cytosine residues. pE194 contained six open reading frames which were theoretically capable of coding for proteins with maximum molecular masses as follows (in daltons): A, 48,300; B, 29,200; C, 14,000; D, 13,900; E, 12,600; and F, 2,700. Insertion of plasmid pBR322 into the single PstI site located in frame A of pE194 resulted in a composite plasmid which could replicate in both Bacillus subtilis and Escherichia coli, suggesting that an intact polypeptide A is dispensable for both replication of pE194 and for MLS resistance. Frame B specified inducible MLS resistance, whereas frame F specified the putative peptide associated with the proposed B determinant translational attenuator. The extent to which frames C, D, and E, all contained in TaqI fragment B, were translated into polypeptide products is not known; however, a base change in frame E was found in a comparison between the high-copy-number mutant, cop-6, and the wild-type strains. PMID- 6279576 TI - Isolation and characterization of angiotensin converting enzyme from human kidney. AB - Angiotensin converting enzyme [EC 3.4.15.1] was solubilized from the membrane fraction of human kidney cortex using trypsin and purified to homogeneity by DEAE cellulose, hydroxylapatite and DEAE-Sephadex A-50 column chromatographies, preparative isoelectric focusing, and Sephadex G-200 gel filtration. The final recovery of the enzyme was 13.9%. The molecular weight of the enzyme was estimated to be 199,000 by a sedimentation equilibrium method. A value of 170,000 was obtained for the reduced and denatured enzyme by dodecylsulfate polyacrylamide gel electrophoresis. The enzyme was a glycoprotein consisting of a single polypeptide chain with an isoelectric point of 5.10. Neutral sugar accounted for 13% per weight of the enzyme. The purified enzyme had a specific activity of 96.9 mumol/min/mg protein for hippurylhistidylleucine. The Km value, Kcat value and hydrolytic coefficient (Kcat/Km) of the enzyme for hippurylhistidylleucine were 2.0 mM, 545 s-1 and 273 mM-1 . s-1, respectively. Rabbit antibody against the human kidney converting enzyme inhibited the activities of the enzymes from human lung and serum as equally as that from human kidney, but not those from sheep, dog, or rat sera. The human kidney and lung converting enzymes were immunologically identical on double immunodiffusion analysis. PMID- 6279577 TI - Studies on the cation channel in sarcoplasmic reticulum vesicles. I. Characterization of Ca2+-dependent cation transport by using a light scattering method. AB - The characterization of the cation channel in sarcoplasmic reticulum (SR) vesicles was performed by measuring the choline influx. The choline influx in SR vesicles was measured by following the change in light scattering intensity using a stopped flow apparatus. From the analysis of the initial rate of choline influx, the following results were obtained. (1) The choline influx was activated by extravesicular Ca2+ with an apparent dissociation constant of 8.3 x 10(-7) M and was inhibited through two steps with inhibition constants of 2.6 x 10(-5) M and 7.4 x 10(-4) M. (2) The dependence of choline influx on the ion concentration followed the Michaelis-Menten kinetics with a half-saturation constant of 30 mM. (3) The choline influx was inhibited by lowering the pH. (4) The activation energy of choline influx was 3.5 kcal/mol. (5) The choline influx was strongly blocked by Cs+ with an inhibition constant of 10 mM. These properties of the choline transporting system in SR vesicles are similar to those of the cation channel reported by Miller (J. Membrane Biol. (1978) 40, 1--23). Lastly, the effect of extravesicular Ca2+ on the choline transport can be explained by an allosteric model. PMID- 6279578 TI - Developmentally regulated glycoprotein alterations in Dictyostelium discoideum. AB - We have studied the changes in glycoprotein patterns on the surface of Dictyostelium discoideum cells during development on a membrane filter and in suspension culture. Cell-surface carbohydrates were detected by a periodate [3H]borohydride method. The appearance and the increase of some glycoproteins during the early developmental stage on a membrane filter were induced 6 h after the onset of starvation. In suspension culture the same proteins appeared about 2 h earlier and less copies per cell were made. Some glycoproteins present in growth-phase cells, especially with high molecular weights, partially disappear during development on a membrane filter. However, the latter changes were not observed in suspension culture. Pulses of cAMP, which generally accelerate cell development, induced only the appearance of one glycoprotein. We conclude that the changes in patterns of plasma membrane glycoproteins during early development are regulated by several factors; starvation, pulses of cAMP, cell-cell contact and transfer onto a membrane filter. PMID- 6279579 TI - Purification and properties of alpha-galactosidase from immature stalks of Saccharum officinarum (sugar cane). AB - The immature sugar cane stalks studied contained less than 7% sucrose, and showed the activities of enzymes such as invertase, alpha-galactosidase, alpha mannosidase, beta-N-acetylglucosaminidase, beta-glucosidase, beta-xylosidase, and beta-galactosidase. The alpha-galactosidase was highly purified by ammonium sulfate fractionation, gel filtration on a Sephadex G-100 column, ionexchange chromatography on DEAE-cellulose, and CM-cellulose columns, and heat treatment (60 degrees C, 15 min) in the presence of 0.2 m D-galactose. In polyacrylamide gel electrophoresis, the purified enzyme was homogeneous, having a molecular weight of approximately 46,000. In gelfiltration, it was approximately 47,000. The activity was optimum at pH 4.5 and at 60 degrees C. The purified enzyme hydrolyzed p-nitrophenyl-alpha-D-galactopyranoside (Km, 0.83 mM; Vmax, 25.0 mumol/mg/min), raffinose (Km, 25.9 mM; Vmax, 15.4 mumol/mg/min), and stachyose (Km, 13.0 mM; Vmax 2.7 mumol/mg/min), in addition to melibiose, guar gum, and locust bean gum. The hydrolysis of p-nitrophenyl-alpha-D-galactopyranoside was markedly inhibited by HgCl2, AgNO3, p-chloromercuribenzoate (PCMB), L-ascorbic acid, melibiose, stachyose, and D-galactose. Also the purified enzyme showed a lectin activity with trypsinized erythrocytes. PMID- 6279580 TI - Molecular multiplicity of nuclease TT1 from Thermus thermophilus HB8. AB - Purified nuclease TT1 from Thermus thermophilus HB8 has multimolecular weight forms, each of which is composed of three different subunits, alpha (10.8 x 10(4)), beta (7.8 x 10(4)), and gamma (4.1 x 10(4)). The molecular weights of this enzyme were estimated by gel filtration, polyacrylamide gel electrophoresis and equilibrium sedimentation. It was found that most of the enzyme has a molecular weight of about 22 x 10(4) being a monomer having the subunit composition of alpha beta gamma. The remaining part of the enzyme has larger molecular weights and is considered to be size-isomers of alpha beta gamma. The alpha-helical content, 5.5--6.5%, and the beta-structure, about 28%, were estimated from the CD spectrum at 4 degrees C. PMID- 6279581 TI - The effect of an in vivo-injected thiol protease inhibitor, E-64-c, on the calcium-induced degeneration of myofilaments. AB - The role of intracellular calcium-dependent proteinase(s) has been investigated in intact rat muscle. When calcium ions were introduced into intact muscle in vitro with ionophore A23187, Z-line loss and concomitant release of alpha-actinin into the medium were observed. The calcium-induced release of alpha-actinin was not diminished in the muscle with in vivo-injection of a thiol protease inhibitor, E-64-c. Intramuscular concentrations of E-64-c were also measured after pulse labeling with [3H]E-64-c followed by subcellular fractionation. Most of the inhibitor was localized in the cytosol, not in the lysosome. Therefore, we conclude that cytosolic as well as lysosomal proteinases in muscle are not inhibited by the in vivo labeling of the protease inhibitor (10 mg/kg). PMID- 6279582 TI - A survey of multiple phosphoprotein phosphatases in cytosols from rat tissues and erythrocytes. PMID- 6279583 TI - Purification of gizzard myosin light-chain phosphatase, and reversible changes in the ATPase and superprecipitation activities of actomyosin in the presence of purified preparation of myosin light-chain phosphatase and kinase. AB - Sepharose 4B conjugated with phosphorylated myosin light chains was used in affinity chromatography of a partially purified preparation of gizzard myosin light-chain phosphatase (MLCP) (Onishi et al. (1979) J. Biochem. 86, 1283-1290). The MLCP preparation thus purified contained, according to SDS gel electrophoresis, three components of 67,000 (67 K), 54,000 (54 K), 34,000 (34 K) daltons. In an accompanying report, Uchiwa et al. (J. Biochem. 91, 273-282 (1982)) described the purification of gizzard myosin light-chain kinase, which consisted of two subunits; 130 K and 17 K daltons. Using the purified preparations of MLCP and MLCK, it was demonstrated a) that reversible changes in the ATPase and superprecipitation activities occur as myosin light chains are enzymatically phosphorylated and dephosphorylated, and b) that addition of a very low concentration of Ca2+ and its removal cause reversible changes in the turbidity of actomyosin suspensions as well as in the state of phosphorylation of myosin light chains only when MLCK and MLCP are both present. These results provide strong support for the proposal (see Ikebe et al. (1977) J. Biochem. 80, 299-302) that MLCK and MLCP play a key role in the Ca2+ regulation in gizzard. PMID- 6279585 TI - Oxygen Enhancement of bactericidal activity of rifamycin SV on Escherichia coli and aerobic oxidation of rifamycin SV to rifamycin S catalyzed by manganous ions: the role of superoxide. AB - Oxygen enhanced the bactericidal activity of rifamycin SV to Escherichia coli K12. Anaerobically grown cells, which had a low level of superoxide dismutase, were more susceptible to the bactericidal activity than aerobically grown cells, which contained a high level of superoxide dismutase. Oxygen also enhanced the inhibition of RNA polymerase activity of rifamycin SV, when Mn2+ was used as a cofactor. Rifamycin S was reduced to rifamycin SV by NADPH catalyzed by cell-free extracts of Escherichia coli K12. These results indicate that the inhibition of bacterial growth by rifamycin SV is due to the production of active species of oxygen resulting from the oxidation-reduction cycle of rifamycin SV in the cells. The aerobic oxidation of rifamycin SV to rifamycin S was induced by metal ions, such as Mn2+, Cu2+, and Co2+. The most effective metal ion was Mn2+. In the presence of Mn2+, accompanying the consumption of 1 mol of oxygen and the oxidation of 1 mol of rifamycin SV, 1 mol of hydrogen peroxide and 1 mol of rifamycin S were formed. Superoxide was generated during the autoxidation of rifamycin SV. Superoxide dismutase inhibited the formation of rifamycin S, but scavengers for hydrogen peroxide and the hydroxyl radical did not affect the oxidation. A mechanism of Mn2+-catalyzed oxidation of rifamycin SV is proposed and its relation to bactericidal activity is discussed. PMID- 6279584 TI - Subcellular localization of O2- generating enzyme in guinea pig polymorphonuclear leukocytes; fractionation of subcellular particles by using a Percoll density gradient. AB - An iso-osmotic Percoll density gradient was applied to determine the subcellular localization of the O2- generating enzyme, NADPH oxidase, in guinea pig polymorphonuclear leukocytes (PMN). [14C]Myristate (MA) was employed as a metabolic stimulant in order to clarify whether the myristate binding site on PMN membrane was identical with the O2- generating site. The distribution pattern of the O2- generating activity of MA-activated PMN was compared with that of unactivated PMN in parallel experiments to find fractions showing an enhanced O2- generating activity (i.e., above the background values due to O2-generation by other electron-transport systems). We observed very high O2- generating activity concentrated in a single peak with MA-activated PMN but little activity was seen with unactivated PMN in the Percoll density gradient. The O2- generating activity of MA-activated PMN was consistently associated with 5'-nucleotidase activity as a membrane marker enzyme, but was not associated with lysosomal marker enzymes such as myeloper-oxidase and lysozyme. O2- generating and 5'-nucleotidase activities in the peak fraction of MA-activated PMN were increased to about six and four times those of whole cells in terms of specific activity, respectively. These results indicate that the O2- generating enzyme is located on PMN plasma membrane. Furthermore, [14C]myristate-binding activity was mainly found in the peak fraction containing O2 - generating enzyme. This suggests that [14C]myristate binds to plasma membrane, and the O2 - generating enzyme may thus be activated. PMID- 6279586 TI - Electron spin resonance studies on the oxidation of rifamycin SV catalyzed by metal ions. AB - The formation of Mn2+- or Cu2+-rifamycin SV complex and the semiquinone radical of rifamycin SV was studied by electron spin resonance spectroscopy. The results indicate that: (1) the complexes Mn2+-rifamycin SV (g parallel = 2.265, g perpendicular = 2.087, A parallel = 177 x 10(-4) cm-1) were formed; (2) in Cu2+ rifamycin SV, Cu2+ was of square-planar type and was ligated to oxygen atoms; (3) the semiquinone radical of rifamycin SV (g = 2.0031) was formed only under aerobic conditions and its formation was inhibited by superoxide dismutase. The formation of hydroxyl radicals during the autoxidation of rifamycin SV was detected by the spin trapping method using N-t-butyl-alpha-phenylnitrone. Superoxide dismutase, mannitol and EDTA inhibited the development of the signal of the spin adduct, indicating that the hydroxyl radical is formed in via the Fenton-type reaction. PMID- 6279587 TI - Counteraction of calcium-activated, phospholipid-dependent protein kinase activation by adenosine 3',5'-monophosphate and guanosine 3',5'-monophosphate in platelets. AB - In intact human platelets activated by thrombin, diacylglycerol is produced with the concomitant disappearance of phosphatidylinositol (PI). This reaction is associated with phosphorylation of a protein having a molecular weight of about 40,000 (40 K protein) and serotonin release. All the reactions are inhibited in a parallel manner by incubation of platelets with either dibutyryl cyclic AMP or 8 bromocyclic GMP, prior to the stimulation by thrombin. The inhibition of these reactions is inversely related to phosphorylation of another group of platelet proteins. Since Ca2+-activated, phospholipid-dependent protein kinase (C-Kinase) is activated by diacylglycerol and is responsible for 40 K protein phosphorylation (Kawahara, Y., Takai, Y., Minakuchi, R., Sano, K., & Nishizuka, Y. (1980) Biochem. Biophys. Res. Commun. 97, 309-317), the results suggest that in platelets both cyclic AMP and cyclic GMP may serve as inhibitors of C-Kinase by counteracting the receptor-linked PI breakdown probably through the actions of cyclic nucleotide-dependent protein kinases. PMID- 6279588 TI - A spin trap study on anaerobic dehalogenation of halothane by a reconstituted liver microsomal cytochrome P-450 enzyme system. AB - In the presence of a spin trapping reagent, N-tert-butyl-alpha-phenylnitrone, anaerobic incubation of halothane (2-bromo-2-chloro-1,1,1-trifluoroethane) with a reconstituted cytochrome P-450 enzyme system of rabbit liver microsomes exhibited an electron spin resonance spectrum containing signals of a nitroxide radical, and all components of the reconstituted system were necessary to produce the nitroxide radical. Formation of 2-chloro-1,1,1-trifluoroethane, the product of anaerobic dehalogenation of halothane by the reconstituted system, was inhibited by N-tert-butyl-alpha-phenylnitrone. These results indicate that a radical intermediate of halothane is produced during the anaerobic dehalogenation reaction, and that the radical is trapped by N-tert-butyl-alpha-phenylnitrone to form a N-tert-butyl-alpha-phenylnitrone radical adduct. PMID- 6279589 TI - A comparison of dephosphorylation of pig glycogen phosphorylase a isoenzymes. PMID- 6279590 TI - Duality of alpha-subunit of canine kidney sodium- and potassium-activated adenosinetriphophatase in electrophoresis. AB - Sodium- and potassium-activated adenosinetriphosphatase (Na+, K+-ATPase) purified from dog kidney outer medulla was examined by polyacrylamide gel electrophoresis and by photoaffinity labeling with N-(ouabain)-N'-(2-nitro-4-azidophenyl) ethylenediamine (NAP-ouabain). The large subunit band (alpha-band) split into two bands on the gel after the enzyme was heat-treated in the presence of 1% sodium dodecylsulfate (SDS). Of the two bands (alpha I and alpha II), alpha I had the same electrophoretic mobility as the original band, while alpha II moved slightly faster. Total conversion into alpha II was not observed, about half of the original remaining as alpha I. Below 60 degree C, heat treatment did not produce alpha II. Phenylmethylsulfonyl fluoride did not prevent the appearance of alpha II. Both alpha I and alpha II were labeled with [3H]NAP-ouabain. Nonspecific incorporation of [3H]NAP-ouabain also occurred irrespective of illumination, but it was removed either by diffusion during staining and destaining of the gel or by treatment of the enzyme with trichloroacetic acid. It is tentatively concluded that the splitting of the band reflects some intrinsic differences in situ of the alpha-subunit of dog kidney membrane Na+,K+-ATPase. PMID- 6279591 TI - A spin label study on human low density lipoprotein. AB - Selective labeling with N-(1-oxyl-2,2,6,6-tetramethyl-4-piperidinyl)-maleimide of human serum LDL has been performed. The spin-labeled LDL exhibited an ESR spectrum containing signals of a strongly immobilized component only. The signals were completely reversible between 4 degrees C and 37 degrees C and fairly stable at each temperature. The spin-labeled LDL which was prepared by the usual method exhibited an ESR spectrum containing signals of both strongly immobilized and weakly immobilized components (5, 6). The latter was unstable above 25 degrees C and changed irreversibly. The strongly binding site showed higher affinity for the nitroxide radical than the weakly binding site, and two kinds of the strongly binding site were demonstrated kinetically. The rate of binding of the nitroxide radical to the two kinds of strongly binding site were estimated to be 4.7 x 10(4) M-1 . day-1 and 0.16 x 10(4) M-1 . day-1 at pH 7.4 and 4 degrees C, respectively. Both the strongly immobilized and weakly immobilized radicals were reduced with ascorbate at the same rate. It was also shown on gel filtration of the SDS-treated LDL derivatives that the strongly immobilized component was on the apoprotein B moiety, whereas either noncovalent binding to LDL or binding to some small molecular species other than protein was suggested for the weakly immobilized component. PMID- 6279592 TI - A 15N-NMR study on ribonuclease T1-guanylic acid complex. AB - Ribonuclease T1 is highly specific for the guanylic acid residue in polyribonucleotides. To clarify the origin of the substrate specificity, the interaction sites of guanylic acid with ribonuclease T1 were investigated by the use of 15N-NMR. 95% 15N-enriched guanosine-3'-phosphate was prepared and mixed with purified ribonuclease T1. 15N-NMR spectra of the mixtures at different concentrations were obtained and compared with that of the 15N-enriched substrate alone. Upon complex formation, a 15N signal assigned to the amino group nitrogen at position 2 of guanine shifted and was significantly broadened, suggesting a strong interaction with the enzyme through the amino group. This observation is consistent with the results of studies on the substrate specificity of chemical modification. Nuclear Overhauser effects of signals assigned to N-7 and N-3 were also changed, but not shift was observed. The observations do not support the occurrence of protonation at N-7 upon complex formation, which was previously proposed. PMID- 6279593 TI - Synthesis of phosphorylated recognition marker in lysosomal enzymes is located in the cis part of Golgi apparatus. AB - Rat liver membranes were subjected to centrifugation in a sucrose density gradient in which the Golgi apparatus was separated into several subfractions. Two enzymes involved in the synthesis of the phosphorylated recognition marker in lysosomal enzymes, UDP-N-acetylglucosamine:lysosomal enzyme precursor N acetylglucosamine-1-phosphotransferase and alpha-N-acetylglucosaminyl phosphodiesterase fractionated with alpha-1,2-mannosidase, a marker enzyme of cis Golgi membranes and differently from galactosyltransferase, a marker enzyme of trans Golgi membranes. PMID- 6279594 TI - Detection of a g' = 1.74 EPR signal in bovine spleen purple acid phosphatase. AB - When purified with hydroxylapatite, bovine spleen purple acid phosphatase, bearing two iron atoms/molecule, is EPR-silent. In contrast, enzyme purified without hydroxylapatite exhibits the distinctive g' = 1.74 EPR signal characteristic of porcine uteroferrin, with an intensity accounting for about 10% of the total iron. The intensity of the signal is increased 8-fold by the addition of ferrous iron. This treatment, while shifting the visible absorption maximum of the protein from 550 to 525 nm, does not significantly alter the intensity of its visible absorption. Loss of the g' = 1.74 EPR signal upon addition of phosphate to EPR-active preparations and the detection of virtually stoichiometric amounts of phosphate in the protein as isolated suggest that phosphate-binding may abolish the g' = 1.75 EPR signal. Such binding may bring the two iron atoms of the enzyme into juxtaposition, causing loss of EPR signal intensity either through spin-lattice relaxation broadening or antiferromagnetic exchange coupling, perhaps involving phosphate or other ligands intercalated between the two paramagnetic iron atoms. PMID- 6279595 TI - Normal and neoplastic human cells have different histone H1 compositions. AB - The H1 histone is the least conserved of the five major groups of histone proteins. There are as many as five subtypes of H1 histone (1-3). These H1 subtypes occur in different amounts in different animal species and also show tissue specificity (1-3). Normal and neoplastic tissues from the same animal, e.g. rat or calf, contain the same H1 subtypes but in different relative amounts (4, 5). Because H1 subtypes exhibit tissue specificity, it is therefore difficult to identify the changes in their composition that are associated with neoplasia reported in studies utilizing tissues of different origins (4, 5). Tissue culture cells and their in vitro transformed neoplastic counterparts, on the other hand, offer an excellent system in which to study these changes because the cells are derived from the same origin. We have examined normal and neoplastic human cells and found a relationship between the H1 composition and the ability of the cells to form tumors in nude mice. The ratio of H1A to H1B in normal human cells is considerably lower than that in neoplastic cells and this ratio increases with the increased ability of the cells to form tumors in nude mice. PMID- 6279596 TI - Insulin-dependent and insulin-independent low Km cyclic AMP phosphodiesterase from rat adipose tissue. AB - Chromatographic analysis of a soluble extract of rat adipose tissue on DEAE Sephacel resolves four distinct peaks of 3':5'-nucleotide phosphodiesterase (EC 3.1.4.17) activity. Kinetic investigation indicates that two of these fractions have a high affinity for cyclic AMP and show negative cooperative kinetic behavior at high substrate concentration. They differ in the degree of inhibition by cyclic GMP and in their response to insulin. If rat epididymal fat pads are incubated with insulin prior to homogenization, only one of the low Km cyclic AMP phosphodiesterase forms is stimulated. PMID- 6279597 TI - Low temperature EPR spectroscopic characterization of the interaction of cytochrome P-450cam with a spin label analog of metyrapone. PMID- 6279598 TI - Ontogeny of the (Na+,K+)-ATPase during chick skeletal myogenesis. PMID- 6279599 TI - Effect of platelet-derived and endothelial cell-derived growth factors on the low density lipoprotein receptor pathway in cultured human fibroblasts. AB - Human platelet-derived growth factor (PDGF) has been previously shown to stimulate low density lipoprotein (LDL) receptor activity in cultured cells. Studies were conducted to delineate in detail the effects of PDGF on the LDL receptor pathway in normal human fibroblasts and to explore relationships between the effects of PDGF on LDL metabolism, on cholesterol metabolism, and on DNA synthesis. Increasing concentrations of PDGF stimulated parallel increases in both DNA synthesis and 125I-LDL cell surface binding. The effect of PDGF was due entirely to an increase (up to 4.3-fold) in the number of receptor sites per cell, and not to a change in receptor affinity (Kd approximately 2.0 nM). Parallel PDGF concentration-dependent increases in 125I-LDL binding, internalization, and degradation at 37 degrees C were observed. The results indicate that PDGF-stimulated cells metabolize receptor-bound LDL in a manner that is identical with that seen with quiescent cells. A single study with highly purified PDGF demonstrated that it was PDGF itself, and not some other component in the partially purified PDGF preparation used in most of this work, that was responsible for the observed effects. Studies were conducted on the effects of PDGF on hydroxymethylglutaryl CoA reductase activity, on cholesterol esterification, and on down-regulation by LDL of the LDL receptor. These studies indicated that LDL cholesterol taken into the PDGF-stimulated cell via the receptor pathway, appeared to become available normally and to have metabolic effects within the cell similar to those seen in quiescent cells. Fibroblasts from subjects with familial hypercholesterolemia showed a normal mitogenic response to PDGF, despite the absence or near absence of an effect on the LDL receptor pathway. Finally, studies were also conducted with endothelial cell conditioned medium (ECCM), used as a source of the endothelial cell-derived growth factor. ECCM was similar to PDGF in stimulating LDL binding, but differed strikingly from PDGF in that the degradation of internalized LDL was inhibited. As a result, ECCM-treated cells did not effectively increase cholesterol esterification or suppress hydroxymethylglutaryl CoA reductase activity when LDL was present. These findings with substances produced by endothelial cells may have important implications for atherogenesis. PMID- 6279600 TI - Direct evidence that bacterial lipopolysaccharides elevate cyclic GMP levels in rat fetal liver cells. AB - We have previously shown that crude bacterial lipopolysaccharide (LPS) preparations markedly increase cGMP levels in rat fetal liver cells in a time- and dose-dependent manner. To provide evidence that this effect was due to LPS and not an impurity in the preparations, three series of experiments were undertaken. First, LPS was prepared from Escherichia coli 055:B5 cells and its cGMP potency assessed at various stages of purification; second, the cGMP activity of three highly purified LPS preparations of known chemical structure was measured, and third, a well characterized LPS was broken into its lipid A and polysaccharide fractions and the cGMP activity of each fraction determined. The results showed that the cGMP stimulatory activity in E. coli 055:B5 cells co purified in a parallel fashion with the LPS molecule derived from those cells, that the three chemically defined, highly purified LPS preparations were all very potent stimulators of cGMP levels, and that the ability to increase cGMP levels of lipid A prepared from a highly purified LPS was comparable in potency to the intact LPS, whereas the polysaccharide portion of the molecule was without activity. These findings indicate that the cGMP effect of LPS preparation is due to LPS and not a contaminant and that the activity resides within the lipid A moiety of the molecule. PMID- 6279601 TI - Rapid temperature-dependent transformation of the thyrotropin-releasing hormone receptor complex in rat pituitary tumor cells. PMID- 6279602 TI - Stimulation of [3H]cIMP binding by cAMP analogs in extracts of perfused rat hearts. PMID- 6279603 TI - Formation of low spin complexes of ferric cytochrome P-450-CAM with anionic ligands. Spin state and ligand affinity comparison to myoglobin. PMID- 6279604 TI - ADP-ribosyltransferase activity in cultured hepatocytes. Interactions with DNA repair. AB - A rapid increase in ADP-ribosyltransferase activity was observed when freshly isolated hepatocytes derived from adult rats were established in primary monolayer culture. (ADP-ribose)n-degrading activity remained constant over a period of 48 h of culture. Inhibition of ADP-ribosyltransferase activity with pyridine derivatives, 3-aminobenzamide, theophylline, or thymidine, was accompanied by an enhanced DNA repair synthesis in response to the direct-acting carcinogen, methyl methanesulfonate, or UV irradiation. Three aminobenzamides differing only in the position of the amino group exhibited the same structure activity relationship in regard to their action on DNA repair synthesis and ADP ribosyltransferase. Spermine treatment of hepatocytes apparently had an inverse effect on both these cellular functions. The removal of DNA strand breaks following methyl methanesulfonate treatment was accelerated by inhibitors of ADP ribosyltransferase. The results suggest that ADP-ribosylation interacts with late stages in the process of DNA repair. This interaction apparently is dependent on the nature of damage imposed on chromatin since repair synthesis in response to a number of carcinogens is unaffected by inhibitors of ADP-ribosyltransferase. PMID- 6279605 TI - Solubilized cytochrome c oxidase from Paracoccus denitrificans is a monomer. AB - Cytochrome c oxidase purified from the bacterium Paracoccus denitrificans was analyzed by analytical ultracentrifugation. In the detergent octyltetra/pentaoxyethylene (C8E45), the isolated enzyme exhibits a molecular weight of 79,000 to 84,000. The detergent-solubilized enzyme is thus a monomer which contains one copy of each of the two subunits. PMID- 6279606 TI - Bovine lung cyclic GMP-dependent protein kinase exhibits two types of specific cyclic GMP-binding sites. AB - Cyclic GMP-dependent protein kinase from bovine lung exhibits two distinct and specific binding sites for cyclic GMP. Cyclic GMP bound to Site II dissociates very rapidly (t 1/2 less than 10 s at 0 or 30 degrees C) and requires the presence of ammonium sulfate in the wash buffer for its detection using cellulose ester filters. Both sites are highly specific for cyclic GMP and show affinities for cyclic GMP in the physiological range (less than 1 microM at 30 degrees C). Only cyclic GMP bound to the high affinity site (Site I) correlated with stimulation of histone kinase activity using arginine-rich histone as the substrate; however, both sites were involved with stimulation of histone kinase activity using mixed histones as the substrate. Stimulation of autophosphorylation by cyclic GMP was closely correlated with the binding of cyclic GMP to Site I. PMID- 6279607 TI - The active site structure of Na+- and K+-stimulated ATPase. Location of a specific fluorescein isothiocyanate reactive site. AB - Fluorescein 5'-isothiocyanate (FITC) has been shown to specifically inactivate the Na+- and K+-stimulated adenosine triphosphatase ((Na,K)-ATPase) at low concentrations (Karlish, S. J. D. (1979) Na+,K+ATPase Structure and Kinetics 115 128). The site of modification of purified dog kidney (Na,K)-ATPase by FITC has been investigated by enzymatic cleavage and fluorescence resonance energy transfer. The binding of FITC, which occurs at a stoichiometry of approximately one site per ATP binding site, causes an ATP-protectable inactivation of ATPase activity suggesting that it is reacting at the ATP hydrolysis site. The FITC reaction site apparently is located near the center of the COOH-terminal 77,000 dalton peptide fragment obtained by chymotryptic cleavage of the alpha subunit. Addition of ouabain to the native enzyme in the presence of chymotrypsin enhances cleavage at this site and releases the fluorescein moiety from the membrane. It is further shown that the distance from the FITC reaction site to the ouabain binding site, as judged by fluorescence resonance energy transfer from anthroyl ouabain to FITC, is approximately 74 A. These results demonstrate that ouabain inhibits the (Na,K)-ATPase by causing a protein conformational change which extends an unusually large distance across the membrane. PMID- 6279608 TI - Na+ transport by the (Na+)-stimulated adenosine triphosphatase. AB - We have previously shown that the (Na+ and K+)-stimulated adenosine triphosphatase from canine kidney reconstituted into phospholipid vesicles is capable of K+-independent transport of Na+ against a concentration gradient (Forgac, M., and Chin, G. (1981) J. Biol. Chem. 256, 3645-3646). The Na+ dependence of the stoichiometry of this process has been investigated and suggests that Na+ ions can be bound and transported by the sites normally occupied by K+. Measurement of the membrane potential generated during active Na+ uptake by [3H]triphenylmethylphosphonium ion distribution gave a value of 50 mV (positive inside). The electrogenicity of Na+ transport is sufficient to explain the observed uptake of an approximately equivalent amount of Cl-. We have also measured Na+ transport into reconstituted vesicles containing neither Na+ nor K+. Na+ uptake was half-maximal at an external Na+ concentration of 1 mM and was accompanied by a less than equivalent amount of Cl- uptake. In order to determine whether protons might also be moving in this system, the intravesicular pH was measured with the pH-sensitive fluorescent probe 1-hydroxypyrene-3,6,8 trisulfonic acid. Active Na+ uptake was accompanied by proton efflux, the amount of proton movement depending on the counterion present. PMID- 6279609 TI - Maximal limits of the Escherichia coli replication factor Y effector site sequences in pBR322 DNA. AB - pBR322 DNA contains two separate regions on opposite strands and close to the origin of replication which, when in single-stranded form, can act as effectors for the ATPase activity of Escherichia coli replication factor Y. Small fragments of DNA containing these sites when cloned into an fl phage vector act as origins of DNA replication allowing the formation of complementary double-stranded DNA in a rifampicin-resistant, dnaB-, dnaG-, and dnaC-dependent fashion in vitro. We report here the maximal limits of the E. coli replication factor Y effector sites of pBR322 DNA. The site on the H strand of pBR322 DNA can form a structure resistant to digestion by E. coli exonuclease VII, the site on the L strand cannot. The H and L strand sites lie within nucleotides 2144-2185 and 2353-2416, respectively, of pBR322 DNA. A deletion of 34 nucleotides (nucleotides 2121-2154) within the H strand site renders it totally inactive as an effector for factor Y ATPase activity. No extensive homology could be detected between the three known factor Y effector sites, the two reported here and the one previously identified on the phi X174 viral DNA strand (Shlomai, J., and Kornberg, A. (1980) Proc. Natl. Acad. Sci. U. S. A. 77, 799-803). PMID- 6279610 TI - Characterization of the free radical of mammalian ribonucleotide reductase. AB - Mouse fibroblast 3T6 cells, selected for resistance to hydroxyurea, were shown to overproduce protein M2, one of the two nonidentical subunits of mammalian ribonucleotide reductase. Packed resistant cells gave an EPR signal at 77 K very much resembling the signal given by the tyrosine-free radical of the B2 subunit of Escherichia coli ribonucleotide reductase. Also, the M2-specific free radical was shown to be located at a tyrosine residue. Of the known tyrosine-free radicals of ribonucleotide reductases from E. coli, bacteriophage T4 infected E. coli and pseudorabies virus infected mouse L cells, the M2-specific EPR signal is most closely similar to the signal of the T4 radical. The small differences in the low temperature EPR signals between these four highly conserved tyrosine-free radical structures can be explained by slightly different angles of the beta methylene group in relation to the plane of the aromatic ring of tyrosine, reflecting different conformations of the polypeptide chain around the tyrosines. The pronounced difference in microwave saturation between the E. coli B2 tyrosine radical EPR signal and the M2 signal could be due to their different interactions with unspecific paramagnetic ions or with the antiferromagnetically coupled iron pair, shown to be present in the E. coli enzyme and postulated also for the mammalian enzyme. A difference in the iron-radical center between the bacterial and mammalian ribonucleotide reductase is also observed in the ability to regenerate the free radical structure. In contrast to the B2 radical, the M2 tyrosine free radical could be regenerated by merely adding dithiothreitol in the presence of O2 to a cell extract where the radical had previously been destroyed by hydroxyurea treatment. PMID- 6279611 TI - Escherichia coli heat-labile enterotoxin. Nucleotide sequence of the A subunit gene. AB - We report the complete DNA sequence of the Escherichia coli elt A gene, which codes for the A subunit of the heat-labile enterotoxin, LT. The amino acid sequence of the LT A subunit has been deduced from the DNA sequence of elt A. The LT A subunit starts with methionine, ends with leucine, and comprises 254 amino acids. The computed molecular weight of LT A is 29,673. The A subunit of cholera toxin (CT A) has been shown to be structurally and functionally related to the LT A subunit. Comparison of the primary structure of LT A with the known partial amino acid sequence of CT A indicates that the 2 polypeptides share considerable homology throughout their sequences. The NH2-terminal regions exhibit the highest degree of homology (91%), while the COOH-terminal region, containing the sole cystine residue in each toxin is less conserved (approximately 52%). Alignment of homologous residues in the COOH-terminal regions of LT A and CT A indicates that a likely site for proteolytic cleavage of LT A is after Arg residue 188. The resulting A2 polypeptide would be 46 amino acids long, would contain a single cysteine residue, and have Mr = 5261. The elt A nucleotide sequence further predicts that the LT A protein is synthesized in a precursor form, possessing an 18-amino acid signal sequence at its NH2 terminus. PMID- 6279613 TI - Activation of a rat liver NAD glycohydrolase by histone. PMID- 6279612 TI - Superoxide radical inhibits catalase. AB - Catalase was inhibited by a flux of O2- generated in situ by the aerobic xanthine oxidase reaction. Two distinct types of inhibition could be distinguished. One of these was rapidly established and could be as rapidly reversed by the addition of superoxide dismutase. The second developed slowly and was reversed by ethanol, but not by superoxide dismutase. The rapid inhibition was probably due to conversion of catalase to the ferrooxy state (compound III), while the slow inhibition was due to conversion to the ferryl state (compound II). Since neither compound III nor compound II occurs in the catalatic reaction pathway, they are inactive. This inhibition of catalase by O2- provides the basis for a synergism between superoxide dismutase and catalase. Such synergisms have been observed in vitro and may be significant in vivo. PMID- 6279615 TI - Hyaluronate degradation in 3T3 and simian virus-transformed 3T3 cells. AB - The cellular control of hyaluronate levels was examined in cultures of simian virus 40-transformed 3T3 (SV3T3) and 3T3 cells which are known to differ in their metabolism of hyaluronate. When [3H]hyaluronate was added to cultures of the two cell lines, four times more ligand was bound per mg of protein by the SV3T3 cells than by the 3T3 cells. Of the bound [3H] hyaluronate, 40% was degraded by the SV3T3 cells to oligosaccharides characteristic of the breakdown of hyaluronate, but only 2% was degraded by 3T3 cells. Hyaluronidase activity was found in the cell layer and medium of the SV3T3 cultures, but was not detectable in 3T3 cells. The SV3T3 enzyme was active only at acidic pH, but at neutral pH the secreted SV3T3 hyaluronidase was thermally more stable then the cell-associated enzyme. In contrast, both cell lines were found to contain similar amounts of beta glucuronidase and beta-N-acetylglucosaminidase activity. We conclude that the elevated capacity of SV3T3 cells to degrade hyaluronate may be partially responsible for their lack of the hyaluronate-containing pericellular coat which is prominent around 3T3 cells. PMID- 6279614 TI - Bacteriophage SP6-specific RNA polymerase. II. Mapping of SP6 DNA and selective in vitro transcription. AB - DNA from bacteriophage SP6 grown on Salmonella typhimurium LT2 is a linear duplex with a unique DNA sequence having a molecular weight of 2.9 x 10(6) (43.500 base pairs). Restriction endonuclease cleavage maps on SP6 DNA for Ava II, Kpn I, Bgl II, Eco RI, and HindIII have been determined. SP6 DNA is transcribed selectively in vitro by Escherichia coli RNA polymerase, predominantly from three strong promoter sites located near the left end of the standard physical map, reading rightward to a termination site near 6,000 base pairs. Transcription in vitro by purified SP6-specific RNA polymerase gives rise to at least 10 discrete RNA species, all of which are read rightward. Promoter sites for these species are located throughout the rightmost 90% of the SP6 DNA molecule, although precise mapping has not yet been carried out. The general form of the SP6 transcriptional map is similar to the T7- and T3-like phages, although no gross sequence homologies are evident from DNA-RNA hybridization experiments. PMID- 6279616 TI - Yeast DNA topoisomerase II. An ATP-dependent type II topoisomerase that catalyzes the catenation, decatenation, unknotting, and relaxation of double-stranded DNA rings. AB - An activity from the yeast Saccharomyces cerevisiae, initially noted for its catalysis of aggregation of covalently closed double-stranded DNA rings in the presence of ATP, has been identified as a type II DNA topoisomerase and is designated yeast DNA topoisomerase II. The formation of the DNA aggregate, which has been shown to be a network of DNA rings that are topologically interlocked, requires the presence of a yeast DNA-binding protein in addition to the topoisomerase. In the absence of the binding protein, yeast DNA topoisomerase II catalyzes decatenation and unknotting of duplex DNA rings and the relaxation of negatively or positively supercoiled DNA. All reactions are ATP-dependent and require Mg(II). Similar to other eukaryotic and phage T4-type II DNA topoisomerases, the yeast enzyme does not catalyze DNA supercoiling under the assay conditions employed. The activity is not sensitive to the gyrase inhibitor nalidixic acid, oxolinic acid, or novobiocin. Coumermycin inhibits the activity, however, at a concentration as low as 5 microgram/ml. PMID- 6279617 TI - Rabbit liver glycogen synthase kinases. Characterization of a protein kinase (PC0.7) able to phosphorylate glycogen synthase and phosvitin. AB - A rabbit liver protein kinase (PC0.7), able to phosphorylate glycogen synthase and phosvitin, has been extensively purified. The enzyme had apparent Mr = 170,000-190,000 as judged by gel filtration and was associated with two major polypeptide species, alpha (Mr = 43,000) and beta (Mr = 25,000). Two other polypeptides, Mr = 38,000 and Mr = 35,000, were also detected. Treatment with trypsin led to an enzyme composed only of polypeptides of Mr = 35,000 and Mr = 25,000. The beta-polypeptide underwent autophosphorylation when incubated with Mg2+ and ATP or GTP. The protein kinase was effective in utilizing both ATP and GTP as the phosphoryl donor (apparent Km values 5-11 microM and 9-19 microM, respectively). The enzyme phosphorylated phosvitin, casein, and glycogen synthase but not histone or phosphorylase and was inhibited by heparin. Phosphorylation of glycogen synthase proceeded to approximately 0.5 phosphate/subunit with little inactivation of the glycogen synthase. The phosphorylation occurred predominantly in a 21,000-dalton CNBr fragment of glycogen synthase that had been previously shown to reside toward the COOH terminus of the molecule. The liver PC0.7 appeared very similar to an analogous enzyme isolated from rabbit muscle (DePaoli Roach, A. A., Ahmad, Z., and Roach, P. J. (1981) J. Biol. Chem. 256, 8955-8962). The present work, therefore, provides a point of contact between the Ca2+ and cyclic nucleotide-independent glycogen synthase kinases of rabbit liver and muscle. PMID- 6279618 TI - Alteration of the regulatory subunit of type 1 cAMP-dependent protein kinase in mutant Y1 adrenal cells resistant to 8-bromoadenosine 3':5'-monophosphate. PMID- 6279619 TI - Identification of the paromomycin-resistance mutation in the 15 S rRNA gene of yeast mitochondria. PMID- 6279620 TI - Rabbit skeletal muscle phosphorylase kinase. Catalytic and regulatory properties of the active alpha gamma delta and gamma delta complexes. PMID- 6279621 TI - Affinity labeling of the protein kinase associated with the epidermal growth factor receptor in membrane vesicles from A431 cells. AB - Epidermal growth factor (EGF), a mitogenic polypeptide hormone, stimulates the phosphorylation of certain endogenous proteins in membrane preparations derived from A431 cells, a human tumor cell line. Membrane vesicles prepared from A431 cells were reacted with 5'-p-fluorosulfonylbenzoyl adenosine (5'-p-FSO2BzAdo). Reaction of the vesicles with 5'-p-FSO2BzAdo results in a time-dependent inhibition of EGF-stimulable protein kinase activity which parallels an increase in incorporation into the vesicles of the 5'-p-sulfonylbenzoyl-[8-14C]adenosine moiety from 5'-p-FSO2Bz[14C]Ado. The primary bands labeled have Mr = 170,000 and 150,000. Labeling of these bands by 5'-p-FSO2Bz[14C]Ado is inhibited by incubation of the membrane vesicles with adenyl-5'-yl imidodiphosphate, an ATP analog. Inactivation of the kinase with N-ethylmaleimide or by heating results in a sharply decreased labeling of the proteins with Mr = 170,000 and 150,000. Proteins of these molecular weights have previously been identified in these cells as the EGF receptor and a degradation product of the receptor. These experiments provide chemical evidence that the EGF receptor and the EGF stimulable kinase are the same protein. PMID- 6279622 TI - Partially oxidized active intermediates in refolding of reduced ribonuclease. AB - The refolding of reduced ribonuclease A has been studied by measurements of enzymatic activity under conditions where the oxidation of thiol groups into disulfide bonds is rather slow. The sensitivity to a treatment by N ethylmaleimide has been used to distinguish between partially and totally oxidized active species. It is found that the first active protein molecules to be formed do not have all of their disulfide bonds. Because they are active, these partially oxidized intermediates probably have very close to native conformation, which they can reach without being trapped in a wrong structure by forming too many incorrect disulfide bonds. The significance of these intermediates to the refolding pathway of reduced ribonuclease is discussed. PMID- 6279623 TI - Insulin acutely increases phospholipids in the phosphatidate-inositide cycle in rat adipose tissue. AB - Administration of insulin in vivo provoked rapid (near maximal in 30 min) increases (65-90%) in the concentrations of phosphatidic acid, phosphatidylinositol, and polyphosphoinositides in rat adipose tissue. Insulin also increased phosphatidylinositol levels in adipocytes incubated in vitro. Dose response relationships suggested that these effects of insulin were physiologically relevant. The enrichment of membranes with acidic phospholipids in the phosphatidate-inositide cycle may play a role in the action of insulin in adipocytes. PMID- 6279624 TI - Oxygen toxicity in Streptococcus sanguis. The relative importance of superoxide and hydroxyl radicals. AB - Streptococcus sanguis, whose growth appears to be independent of the availability of iron, makes no hemes, contains neither catalase nor peroxidase, and can accumulate millimolar concentration levels of H2O2 during aerobic growth. It possesses a single manganese-containing superoxide dismutase whose concentration can be varied over a 50-100-fold range by manipulating the availability of oxygen during growth. Cell extracts contain a soluble NADH-plumbagin diaphorase which mediates O2- production in vitro and presumably also in vivo. Plumbagin increased oxygen consumption by S. sanguis and imposed an oxygen-dependent toxicity. Cells grown aerobically and containing elevated levels of superoxide dismutase were resistant to this toxicity. Dimethyl sulfoxide, which was shown to permeate S. sanguis freely, was used as an indicating scavenger of OH. An in vitro enzymic source of O2- plus H2O2 generated formaldehyde from dimethyl sulfoxide, an indication of OH. production. Either superoxide dismutase or catalase inhibited this OH. production and iron salts augmented it. Intact, aerobic cells of S. sanguis also gave evidence of OH. production, in the presence of plumbagin, but all of it appeared to be generated outside the cells. In addition, 0.5 M dimethyl sulfoxide did not diminish the oxygen-dependent toxicity of plumbagin. We conclude that, in S. sanguis, O2- can exert a toxic effect independent of the production of OH.. PMID- 6279625 TI - Functional relationship of the cytochrome b to the superoxide-generating oxidase of human neutrophils. AB - A subcellular particulate fraction containing the NADPH-dependent O2.--generating oxidase from stimulated human neutrophils was prepared. This fraction was depleted of certain enzyme markers of primary and secondary granules and was devoid of measurable myeloperoxidase, both enzymatically and spectrally. When prepared from neutrophils which had been previously stimulated with phorbal myristate acetate, this fraction contained cyanide-insensitive, pyridine nucleotide-dependent O2.--generating activity with a specific activity of 260 nmol min-1 mg-1. O2.--generating activity is completely ablated by p chloromercuribenzoate exposure. Preparations from normal unstimulated neutrophils or stimulated neutrophils from a male patient with chronic granulomatous disease had negligible amounts of this O2.--generating enzymatic activity. The dominant chromophore in this preparation was a b-type cytochrome, the spectral and functional characteristics of which are further described herein. Pyridine nucleotide-dependent reduction of the intrinsic cytochrome b closely parallels O2.- generation in this preparation. Specifically, reduction occurs in preparations from phorbal myristate acetate-stimulated neutrophils and is absent in unstimulated or stimulated p-chloromercuribenzoate-inactivated preparations. PMID- 6279626 TI - Hormonal regulation of the phosphorylation of ATP citrate lyase in 3T3-L1 adipocytes. Effects of insulin and isoproterenol. PMID- 6279627 TI - Interaction of platelet-derived growth factor with its fibroblast receptor. Demonstration of ligand degradation and receptor modulation. AB - Platelet-derived growth factor (PDGF) has previously been shown to bind to a specific high affinity receptor on human foreskin fibroblasts. The present study was carried out to characterize some of the cellular events resulting from the interaction of the ligand with its receptor. Radiolabeled PDGF was rapidly internalized and degraded after binding to the cells. The degradation was complete and was inhibited by low concentrations of the lysosomotropic agents, chloroquine, ammonium chloride, or methylamine, suggesting that the degradation occurs in the lysosomes. The cellular binding capacity for PDGF decreased after exposure of the cells to PDGF at 37 degrees C. This down regulation of the PDGF receptor was optimal after a 60-min incubation at 37 degrees C and half-maximal at 0.5 nM concentration of PDGF. The binding capacity was restored when the PDGF containing medium was changed to medium without PDGF; the binding capacity increased from 40 to 80% od the initial value after a 4-h incubation at 37 degrees C. The reappearance on the cell surface of PDGF-binding sites was dependent on protein synthesis and totally blocked by cycloheximide (20 micrograms/ml). Thus, either the receptor has to be resynthesized after internalization or, alternatively, any step in the recycling of "used" receptors is dependent on protein synthesis. Exposure of the cells to PDGF also caused a dose-dependent decrease in the binding capacity for epidermal growth factor which has a distinct receptor on these cells. In contrast, epidermal growth factor did not modulate the PDGF binding capacity, lending no support to the idea that the receptors for epidermal growth factor and PDGF are processed in a common pathway. PMID- 6279628 TI - The endocytotic rate constant. A cellular parameter for quantitating receptor mediated endocytosis. PMID- 6279629 TI - Kinetics of internalization and recycling of the asialoglycoprotein receptor in a hepatoma cell line. PMID- 6279630 TI - Decreased purine synthesis during amino acid starvation of human lymphoblasts. AB - Normal human lymphoblasts starved for each of several essential, but not essential, amino acids had decreased DNA and RNA synthesis but no change in free intracellular purine nucleotides. The rates of purine nucleotide synthesis via the de novo and salvage pathways were measured by incorporating [14C]formate and [14C]hypoxanthine labels, respectively, into lymphoblasts starved for an amino acid or treated with a protein synthesis inhibitor. After 3 h of starvation, purine synthesis via the de novo pathway decreased 90% and via the salvage pathway decreased 60%. Cycloheximide and puromycin each reduced de novo synthesis by 96% and salvage synthesis by 72%. The decrease in purine synthesis de novo after removal of the amino acid was of first order kinetics and was fully and rapidly reversed by reconstitution with the amino acid. The synthesis of alpha-N formylglycinamide ribonucleotide declined 97% after amino acid starvation; the synthesis of purines from 5-aminoimidazole-4-carboxamide riboside decreased 41%. The synthesis of guanylates decreased more than the synthesis of adenylates during amino acid starvation. PMID- 6279631 TI - Exposure of thymocytes to a low temperature (4 degrees C) inhibits the onset of their hormone-induced cellular refractoriness. AB - The hormonal response of viable mouse thymocytes is radically dependent of their ambient temperature. While at 37 degrees C the cells respond to isoproterenol by an abrupt rise (within 30 s) followed by a exponential decline in the level of intracellular cAMP, at 4 degrees C the level of cAMP remains high, i.e. there is an inhibition of the hormone-induced refractory state. These distinctly different patterns of response are reflected also in both the state of activation of cAMP dependent protein kinase and the activity of adenylate cyclase. The inhibition of cellular refractoriness in the cold is shown to be fully reversible, lasting only as long as the hormone is present in the extracellular medium. Washing out the hormone or displacing it by a specific antagonist (propranolol) results in a decline of cAMP, of the activity ratio of the kinase, and of the activity of the adenylate cyclase back to basal values. Evidence is presented to show that at 4 degrees C there is no significant hormone-dependent decreases in cAMP degradation or efflux. On the other hand, the activity of adenylate cyclase remains persistently high, through neither the hormone-binding site of the receptor nor the active site of the catalytic subunit of the cyclase seem to be impaired. The different response pattern observed at 4 degrees C appears, therefore, to be associated with the transfer and the signal between these two sites and probably with the G/F protein (s). The possibility to dissect in a selective and reversible manner the process of hormonal stimulation (coupling) from the process of desensitization, which, under normal physiological conditions constitute consecutive and inseparable chain of events, leads us to a propose that the signal transfer which enables activation of adenylate cyclase is, somewhere along its way, distinct from the signal transfer which brings about the onset of the refractory state, and to conclude that these two processes are partially autonomous and regulated by either two different proteins or two different sites on the same protein. The postulated proteins (or sites) should, therefore, differ in their sensitivity to temperature changes, a difference which may be most useful in the identification and isolation of the molecular species involved and in the study of their properties and their mechanism of action. PMID- 6279632 TI - Decreased alpha 1-adrenoceptor responsiveness and density in liver cells of thyroidectomized rats. AB - The effects of hypothyroidism on the hepatic alpha 1-receptor system were studied in isolated rat liver cells. Phenylephrine and vasopressin caused concentration dependent activation of glycogen phosphorylase and release of 45Ca from 45Ca loaded cells in either normal or thyroidectomized rats. However, the magnitude of both responses to phenylephrine was markedly suppressed after thyroidectomy and could be restored to near normal levels by in vivo treatment with 1 triiodothyronine (0.25 mg/kg/day) for 4 days. The potency of vasopressin to induce phosphorylase activation and 45Ca release was only slightly reduced by thyroidectomy. Binding of [3H]prazosin to putative alpha 1-receptors in purified liver plasma membranes revealed that the above changes were accompanied by a decrease in the density of binding sites from 567 +/- 51 fmol/mg of protein in controls to 326 +/- 51 fmol/mg in thyroidectomized rats and a return to 498 +/- 23 fmol/mg in thyroidectomized rats treated with 1-triiodothyronine. The affinity of binding sites for [3H]prazosin or for alpha-receptor agonists was the same in the three groups of rats and affinity for epinephrine was unaffected by the presence of guanyl-5'-yl imidodiphosphate (30-100 microM). From these findings, it appears that a reduction in the number of hepatic alpha 1-receptors is responsible for the selective decrease in alpha-adrenergic responses in the hypothyroid rat liver. These changes are opposite to those previously reported for hepatic beta-receptors. PMID- 6279633 TI - Analysis of neurotoxin and mitogen-stimulated sodium transport in human fibroblasts. AB - Human fibroblasts contain two classes of ouabain-insensitive Na+ channels. One channel is activated by alkaloid neurotoxins in combination with scorpion venom, a response which is similar to that of the voltage-sensitive Na+ ionophore of nerve. The other channel is activated by fetal calf serum, several other mitogenic agents, or low extracellular Ca2+. The nervelike Na+ channel is inhibited by tetrodotoxin but not by amiloride, while the mitogen-stimulated channel is blocked by amiloride but not by tetrodotoxin. The combined effects of neurotoxins and low Ca2+ on Na+ uptake are additive, consistent with the conclusion that these Na+ pathways are different and independent. Studies of Na+ efflux show the presence of two intracellular Na+ compartments. When Na+ flux through the (Na+ + K+)-ATPase pump is inhibited by ouabain, compartment A has a high rate constant of efflux (K1 = 0.30 mn-1) and constitutes 60% of the total cellular Na+. Compartment B, with a markedly smaller efflux constant (k2 = 0.07), contains the remaining 40% of cell Na+. Stimulation of flux through the nervelike channel has a pronounced effect on the rate constants and Na+ content of both compartments (K1 = 0.56, k2 = 0.10), but activation of the mitogenic channel has a pronounced effect only on the rate of efflux from compartment A and on the size of compartment B. A cytoplasmic-nuclear model for the Na+ compartments is presented. PMID- 6279634 TI - The mechanism of the insulin-like effects of ionic zinc. AB - The insulin-like effects of ionic zinc (Zn2+) were studied in isolated rat adipocytes. Concentrations of Zn2+ between 250 and 1000 microM stimulated 3-O methylglucose transport and glucose metabolism to CO2, glyceride-fatty acid, and glyceride-glycerol. Selective stimulation of the pentose phosphate cycle was observed since a Zn2+-induced increase in glucose carbon 1 oxidation persisted even when glucose transport was blocked with 50 microM cytochalasin B or when transport was no longer rate-limiting for metabolism at high concentrations of glucose. Enhanced pentose phosphate cycle activity may have been due to a selective inhibition of glutathione reductase by the ion, which was also accompanied by a fall in cellular glutathione content. Zn2+ also inhibited lipolysis stimulated by the beta-adrenergic agent ritodrine in the absence of glucose. The effects of Zn2+ on glucose oxidation and stimulated rates of lipolysis were inhibited by extracellular catalase, indicating that they were largely a result of H2O2 generation. The H2O2 production appeared for the most part to be caused by zinc-catalyzed autoxidation of sulfhydryl groups present on external cell membranes, although involvement of sulfhydryl groups on bovine serum albumin in the buffer could also have contributed. The insulin-like effects of Zn2+ in adipocytes are therefore caused not only by direct effects of the ion on intracellular metabolism but also by indirect effects related to H2O2 generation. PMID- 6279635 TI - The asymmetric distribution of charges on the surface of horse cytochrome c. Functional implications. AB - The electric potential field around native horse cytochrome c and 12 singly modified 4-carboxy-2,4-dinitrophenyl- (CDNP) lysine cytochromes c is asymmetric, mainly because of the inhomogeneous distribution of negative charges. Dipole moments of 325 and 308 debye, (1.08.10(-27) and 1.03.10(-27) coulomb.meter), respectively, were calculated for horse ferri- and ferrocytochrome c. The angle between the heme plane and the dipole vector of horse ferricytochrome c is 33 degrees and increases 1 degree upon reduction to the ferrous form. Dipole moments of the CDNP-lysine cytochromes c differ from that of native cytochrome c by as much as 140 debye in magnitude and 45 degrees in direction. It is proposed that its dipole moment causes cytochrome c to orient itself in the electric fields of its redox partners, and that the CDNP-lysine cytochromes c, which have different dipole moments, do not form a productive complex. Reorientation to the correct position for electron transfer increases the activation energy and lowers the rate of reaction. This model describes quantitatively the relative activities of those CDNP-lysine cytochromes c which are modified outside of the interaction domain and it allows correction of the activities of those modified inside the domain, on the front surface of the molecule, for the change in dipole moment. The interaction domain for the reaction with cytochrome c reductase includes in decreasing order of involvement lysines 13, 72, 86, 27, and 87. That for the reaction with cytochrome c oxidase is slightly smaller, with lysines 13, 72, 86, and 27. The cytochrome c peroxidase domain is the largest of all and is defined by lysines 72, 86, 13, 87, 27,, and 73. All refined interaction domains encompass the exposed heme edge and are to a large extent overlapping, indicating that electron transfer takes place at or close to this prosthetic group and that cytochrome c must move on the outer surface of the inner mitochondrial membrane during electron transport between reductase and oxidase. For a quantitative description of the electrostatic interaction of cytochrome c with other molecules, it is essential to take into account the totality of its charge configuration. PMID- 6279636 TI - Similar glycopeptides in normal chondroblasts and in Rous sarcoma virus transformed fibroblasts. PMID- 6279637 TI - The effect of dexamethasone on the initiation of beta-globin gene transcription in differentiating Friend cells. AB - Plasmid subclones of the 5' and 3' regions of the beta major-globin gene have been used as probes to assay the level and character of the transcripts of this gene in differentiating Friend erythroleukemia cells treated with dexamethasone. Using an S1 nuclease protection protocol, it is shown that dexamethasone reduces the amount of precursor-specific sequences in differentiating Friend cells within 30 min without significantly affecting the nuclear turnover of these molecules. The sites of initiation and termination of transcription remained the same during the shutdown of transcription by the hormone. In contrast to the situation in Friend cells, dexamethasone failed to inhibit globin gene transcription in three normal erythroid tissues. The manner in which the Friend virus complex might render the globin genes sensitive to glucocorticoids is discussed. PMID- 6279638 TI - Purification and properties of Oxidized and reduced forms of Hepatic fructose 1,6 bisphosphatase. AB - D-Fructose 1,6-bisphosphate 1-phosphohydrolase (EC 3.1.3.11) was isolated from rat liver in two forms: "A," isolated in the presence, and "B," isolated in the absence of dithiothreitol. Both forms had an apparently identical molecular weight of approximately 37,000/subunit and the same Km for fructose 1,6 bisphosphate of 2 microM. However, the Ki of the AMP inhibition of form A was 140 microM and of form B, 370 microM. With form B the same inhibition as with form A was reached by incubating the enzyme with dithiothreitol. The two forms of the enzyme differed in their total, as well as in their number of fast reacting thiol groups. Form A was the more reduced form, exhibiting 22.4 thiol groups/molecule, 2.5 of them fast reacting with 5,5'-dithiobis-(2-nitrobenzoic acid). Only 0.5 fast reacting groups and a total of 19.2 were found with form B. The fast reacting thiol groups disappeared when assayed in the presence of AMP. It is suggested that a redox reaction alters a site that influences the inhibitory action of AMP, so as to regulate the activity of fructose 1,6-bisphosphatase. PMID- 6279639 TI - Ovalbumin utilizes an NH2-terminal signal sequence. AB - In synchronized translation experiments in the wheat germ and reticulocyte lysate systems, ovalbumin (385 amino acids) was glycosylated by and segregated in dog pancreatic microsomes only if microsomes were added before the nascent ovalbumin polypeptide contained less than 150 amino acids. This would place the "signal" sequence of ovalbumin prior to residue 150, in contrast to a previous report. PMID- 6279640 TI - Genome-length copies of poliovirion RNA are synthesized in vitro by the poliovirus RNA-dependent RNA polymerase. AB - A soluble RNA-dependent RNA polymerase was purified from the cytoplasm of poliovirus-infected HeLa cells. A single virus-specific protein designated as p63 (or NCVP4) copurified with this activity. The purified polymerase was free of ribonuclease activity and was shown to copy poliovirion RNA when oligo(U) was added to the in vitro reaction mixture. Characterization of the product RNA by electrophoresis in methylmercury (II) hydroxide-agarose gels showed that genome sized copies of poliovirion RNA were synthesized in vitro by the purified polymerase. The product RNA was shown to be heteropolymeric, complementary to virion RNA, and covalently linked to oligo(U). The product RNA contained the expected distribution of UMP and GMP containing dinucleotide pairs which included a very low frequency of CpG pairs. The amount, size distribution, and rate of synthesis of product RNA was very dependent on the in vitro reaction conditions. Full sized product RNA was synthesized in about 6 min when reaction conditions were used that yielded maximum elongation rates (pH 8.0, 7 mM Mg2+, 37 degrees C). Under these conditions, most of the product RNA recovered from a 1-h reaction was full sized. Thus, the polymerase was found to specifically initiate synthesis at the 3'-end of the template using an oligo(U) primer and to carry out an elongation reaction at about 1250 nucleotides/min that resulted in the synthesis of full sized product RNA. PMID- 6279641 TI - Regulation of cyclic adenosine 3':5'-monophosphate phosphodiesterases. Interrelationship of the various forms in rat skeletal myoblasts and adult muscle. PMID- 6279642 TI - Metallothionein accumulation may account for intracellular copper retention in Menkes' disease. AB - Cultured lymphoblasts derived from infants with Menkes' disease exhibit the same increased avidity for copper as do fibroblasts and most extrahepatic tissues from these patients. The Menkes' cells preferentially take up not only copper but also, on exposure to elevated metal concentrations, the other metallothionein binding metals, zinc and cadmium. Menkes' lymphoblasts contain larger amounts of metallothionein than normal cells following exposure to each of these metals; the amount bound to this protein quantitatively accounted for the total cellular increment in metal in Menkes' cells. Induction of metallothionein synthesis caused both normal and Menkes' cells to subsequently take up increased amounts of 67Cu. These observations suggest that an enhanced capacity of Menkes' cells to accumulate metallothionein may be responsible for their increased uptake and retention of copper. PMID- 6279643 TI - Phosphorylation of high mobility group 14 protein by cyclic nucleotide-dependent protein kinases. AB - Chromosomal high mobility group (HMG) proteins have been examined as substrates for cGMP-dependent and cAMP-dependent protein kinases. Of the four HMG proteins only HMG 14 contained a major high affinity site which could be phosphorylated by both enzymes, preferentially by cGMP-dependent protein kinase. One mol of 32P was incorporated/mol of HMG 14. Kinetic analysis revealed apparent Km and Vmax of 40.5 microM and 14.7 mumol/min/mg, respectively, for cGMP-dependent protein kinase, and 123 microM and 11.1 mumol/min/mg, respectively, for cAMP-dependent protein kinase. Tryptic maps of 32P-labeled phosphopeptides of HMG 14 demonstrated phosphorylation of the same site by both enzymes. The tryptic fragment containing the major phosphorylation site was identified by amino acid composition and sequence as HMG 14 (residues 4-13): H-Lys-Val-Ser(P)-Ser-Ala-Glu Gly-Ala-Ala-Lys-OH. HMG 14 and HMG 17 also contained minor sites which could be phosphorylated by cGMP-dependent protein kinase. Tryptic phosphopeptides mapping suggested that the same minor site was phosphorylated on both HMG 14 and 17. On the basis of amino acid composition, the tryptic peptides carrying the minor phosphorylation sites were identified as H-Leu-Ser(P)-Ala-Lys representing residues 23-26 and 27-30 of HMG 14 and HMG 17, respectively. PMID- 6279644 TI - Tandem duplication and multiple functions of a receptor gene in bacterial chemotaxis. AB - The aspartate receptor in bacterial sensing, previously identified with the tar gene, has been shown to be duplicated in tandem in Escherichia coli. Each gene, which we refer to as tar and tap, respectively, codes for a 60,000-dalton protein. By genetic engineering experiments in which each gene is introduced separately into E. coli strains, it is shown that each transmembrane receptor can respond to the small molecule aspartate, to the maltose-protein-chemoeffector complex, and to repellents. PMID- 6279646 TI - A mutation in the rnh-locus of Escherichia coli affects the structural gene for RNase H. Examination of the mutant and wild type protein. AB - Ribonuclease H (RNase H, EC 3.1.26.4) was purified to homogeneity from Escherichia coli wild type strain KS 351 and the RNase H mutant strain FB 2. The specific activity of the wild type enzyme was 43,200 units/mg, while that of the mutant enzyme was 3,430 units/mg, less than 8% of the wild type activity. Isoelectric focusing also revealed differences in the protein from mutant and wild type. The activity of the wild type enzyme was separated into two peaks with isoelectric points of 9.6 and 9.0. In contrast, the activity of the mutant enzyme focused in a single peak with a pI of 9.4. These results indicate that the mutation in the FB2 strain affects the structural gene for RNase H. The molecular weight of both enzymes was determined by gel filtration as well as NaDodSO4 polyacrylamide gel electrophoresis and found to be identical. Both enzymes are very sensitive to increased temperatures and show indistinguishable rates of inactivation. The basis for the heterogeneity of the isoelectric point and the altered activity of the mutant enzyme is still unknown. PMID- 6279645 TI - Albumin stabilizes leukotriene A4. AB - Chemical analysis of intact leukotriene A4 showed that vertebrate albumins prolonged its aqueous half-life. At pH 7.4, leukotriene A4 hydrolyzed by first order reaction kinetics with rate constants inversely proportional to the albumin concentration. The stabilizing effect of albumin varied quantitatively among different species. Certain agents, such as warfarin, that interact with the site I binding region of albumin reversed its stabilizing effect. Sequestration and exposure of leukotriene A4 to a hydrophobic, alkaline microenvironment of albumin would account for the results. The amino acid sequences Lys-Ala-Trp-Ala-Val-Ala Arg from residues 211-217 of human albumin or Lys-Ala-Trp-Ser-Val-Ala-Arg from residues 210-216 of bovine albumin are compatible with this requirement. The persistence of leukotriene A4 in the presence of albumin confirms and extends our recent observations on its uniform and predictable influence on eicosanoid stability. The significance of this influence is uncertain; however, albumin can no longer be viewed as inert considering its capacity to modify the stability of several, structurally diverse eicosanoids. PMID- 6279647 TI - Leukotriene B4 and phosphatidic acid are calcium ionophores. Studies employing arsenazo III in liposomes. PMID- 6279648 TI - The free radical formed during the hydroperoxide-mediated deactivation of ram seminal vesicles is hemoprotein-derived. AB - Prostaglandin synthase is a multi-enzyme complex which catalyzes the oxygenation of arachidonic acid to the various prostaglandins. During the oxygenation, the enzyme is self-deactivated and, on the basis of ESR data, it has been proposed to form a self-destructive free radical. The free radical was suggested to form from the oxygen lost from prostaglandin G2 during its reduction to prostaglandin H2, and the destructive species was therefore thought to be an oxygen-centered free radical, tentatively identified as the hydroxy radical. We have reinvestigated this ESR signal (g = 2.005) and have concluded, with the aid of the known ESR parameters for the hydroxy and other oxygen-centered free radicals, that the free radical formed during the oxygenation is neither a hydroxy nor any known oxygen centred radical. Prostaglandin synthase is thought to be a hemoprotein, so this unknown ESR signal was compared with the previously observed free radical formed by the reaction of H2O2 with methemoglobin. This comparison indicates that the free radical formed by the reaction of prostaglandin G2 with ram seminal vesicles is hemoprotein-derived and may be formed by the oxidation of an amino acid(s) located near the iron of the heme. PMID- 6279649 TI - Kinetic aspects of hemoglobin.haptoglobin-receptor interaction in rat liver plasma membranes, isolated liver cells, and liver cells in primary culture. AB - 125I-Hemoglobin.haptoglobin injected intravenously into rats was incorporated into liver parenchymal cells as evidenced by a cell separation technique. A mixture of freshly isolated liver parenchymal and nonparenchymal cells failed to internalize and degrade the 125I-hemoglobin.haptoglobin added, although it retained the ability to bind the molecule. The liver parenchymal cells in primary culture also lacked the ability to degrade 125I-hemoglobin.haptoglobin, although they bound the molecule more extensively as compared with the freshly isolated liver cells. It was confirmed that the 125I-hemoglobin.haptoglobin which was bound to the freshly isolated liver parenchymal cells localized on the outer surface of liver plasma membranes. Scatchard plots revealed the existence of two binding sites for 125I-hemoglobin-haptoglobin on the isolated liver plasma membrane: an apparent high affinity binding site (Kd = 1.3 X 10(-7) M) and an apparent low affinity binding site (Kd = 4.0 X 10(-6) M) at 37 degrees C. In contrast, freshly isolated liver parenchymal cells had only an apparent low affinity binding site (Kd = 1.4 X 10(-6) M) at 37 degrees C. Impairment of the apparent high affinity binding site during the isolation procedure with collagenase seemed to be related to loss of the ability to internalize and degrade the 125I-hemoglobin.haptoglobin molecules into the freshly isolated liver parenchymal cells or liver parenchymal cells in primary culture. PMID- 6279650 TI - Synthetic peptide substrates for a tyrosine protein kinase. AB - Immunoprecipitates containing the transforming protein of the avian sarcoma virus, Y73, together with its associated tyrosine-specific protein kinase, have an activity which will phosphorylate the synthetic peptide Lys-Leu-Ile-Glu-Asp Asn-Glu-Tyr-Thr-Ala-Arg at the tyrosine residue. This peptide corresponds to 10 out of 11 amino acids surrounding the phosphorylated tyrosine in both pp60src and P90, the transforming proteins of Rous sarcoma virus and Y73 virus, respectively. The apparent Km for phosphorylation of the peptide was about 5 mM. A second peptide with the sequence Lys-Leu-Ile-Asp-Asn-Glu-Tyr-Thr-ala-Arg differing from the first peptide only by the absence of the glutamic acid 4 residues from the tyrosine was also phosphorylated, but the apparent Km for the reaction was 40 mM. Several sites of tyrosine phosphorylation in viral transforming proteins have been found to have one or more glutamic acids close to the phosphorylated tyrosine on the NH2-terminal side. Taken together with our in vitro phosphorylation studies, this suggests that the primary sequence surrounding target tyrosines may play a role in recognition of substrates by tyrosine protein kinases, and in particular, that glutamic acid residues on the NH2-terminal side may be important. PMID- 6279651 TI - Nucleoside phosphotransferase from barley. Characterization and evidence for ping pong kinetics involving phosphoryl enzyme. AB - A nucleoside phosphotransferase has been purified to homogeneity from barley seedlings. Its Mr is about 50,000 and it consists of two subunits of equal size. A tightly bound metal ion required for activity can be replaced by Mg, Ba, or Co ions, but not by divalent Cu, Ca, Cd, or Hg. The enzyme is capable of catalyzing the transfer of phosphate from nucleoside monophosphates to the 5'-hydroxyl of any other nucleoside, but shows a decided preference for purine deoxynucleoside phosphate acceptors. A short lived phosphoryl enzyme intermediate has been trapped by rapid denaturation in presence of [32P]AMP. The steady state kinetics of this enzyme is found to be fully consistent with a branched ping pong pathway involving a compulsory phosphoryl enzyme intermediate from which phosphate may be transferred to nucleoside or to water. With appropriately chosen substrates, parallel line reciprocal plots were obtained, provided that both products of the branched pathway were taken into account in the rate measurements. Conversely, when only transphosphorylation was measured, converging reciprocal plots were obtained. Accordingly, assays were devised for the measurement of three velocity components: transphosphorylation, hydrolysis, and the sum of both reactions. Competitive inhibition of the transferase reaction by higher concentrations of either substrate is attributed to deadend interactions. Instead of being inhibited by the formation of a putative E-P.AMP complex, however, the phosphatase reaction is accelerated almost 2-fold. PMID- 6279652 TI - The effects of copper depletion on structural aspects of cytochrome c oxidase. AB - The removal of copper from beef heart cytochrome c oxidase by either dialysis against potassium cyanide or by treatment with bathocuproine sulfonate produced changes in the enzyme which are indicative of a spin state transition. In the Soret region of the CD spectrum copper depletion of the enzyme caused a significant decrease in amplitude in combination with a red shift of the peak maximum for oxidized samples, while reduced copper-depleted samples exhibited decreased amplitude and a blue shift of the peak maximum. In the magnetic CD spectra of oxidized copper-depleted samples the peak at 420 nm was shifted to lower wave-length along with a significant increase in amplitude. In reduced samples the peak at 446 nm exhibited a slight red shift concomitant with a substantial decrease in amplitude. The conformational changes indicated by the CD and magnetic CD spectra when copper is removed from the enzyme were supported by the EPR spectra of the NO complex of the reduced copper-depleted enzyme. The removal of copper from cytochrome c oxidase caused the NO complex to exhibit a 3 line splitting pattern of gz in the EPR spectrum instead of the 9 lines seen in the NO complex of the native enzyme. When [15N]NO was used, a 2-line pattern was seen at gz when copper was removed from the enzyme. The changes in the CD and magnetic CD spectra and in the EPR spectra of the NO derivatives of cytochrome c oxidase can be explained by the rearrangement of the axial ligands to iron in cytochrome a3 as a result of copper depletion. These results emphasize the close structural interdependence of the metallic components of this enzyme. PMID- 6279653 TI - The hepatic angiotensin II receptor. I. Characterization of the membrane-binding site and correlation with physiological response in hepatocytes. PMID- 6279654 TI - The hepatic angiotensin II receptor. II. Effect of guanine nucleotides and interaction with cyclic AMP production. AB - Guanine nucleotides were observed to modify the binding of 125I-angiotensin II to rat hepatic plasma membrane receptors. GTP and its nonhydrolyzable analogues greatly increased the dissociation rate of bound 125I-angiotensin II and altered hormone binding to the receptor under equilibrium conditions. In the absence of GTP, 125I-angiotensin II labeled both high affinity sites (Kd1 = 0.46 nM, N1 = 650 fmol/mg) and low affinity sites (Kd2 = 4.1 nM, N2 = 1740 fmol/mg). In the presence of guanine nucleotides, the affinities of the two sites were unchanged, but the number of high affinity sites decreased markedly to 52 fmol/mg. In analogous experiments using the angiotensin II antagonist, 125I-sarcosine1,Ala8 angiotensin II (125I-saralasin), guanine nucleotides minimally affected the interaction of 125I-saralasin with its receptor, increasing the dissociation rate 1.9-fold and the Kd 1.4-fold. The guanine nucleotide inhibition of agonist binding required a cation such as Na+ or Mg2+, with a maximal effect occurring at about 1 mM Mg2+. In liver plasma membranes prepared in EDTA, angiotensin II inhibited basal and glucagon-stimulated adenylate cyclase activities by 30% and 10%, respectively. Angiotensin II also caused a 40% inhibition of glucagon stimulated cyclic AMP accumulation in intact hepatocytes, with a half-maximal effect occurring at 1 nM. The inhibition by angiotensin II of adenylate cyclase in membranes and of cAMP levels in intact cells could be reversed by the antagonist sarcosine1,Ile8-angiotensin II. Vasopressin caused a smaller 26% inhibition of glucagon-stimulated cyclic AMP accumulation. The ability of angiotensin II to inhibit cyclic AMP synthesis may provide an explanation for the observed effects of guanine nucleotides on 125I-angiotensin II binding to plasma membranes. PMID- 6279655 TI - Mechanism of autooxidation for hemoglobins and myoglobins. Promotion of superoxide production by protons and anions. AB - Several hemoglobins and bovine myoglobin are shown to undergo autooxidation reactions promoted by anions. The reduced protein in the presence of oxygen and anion yields the anion complex of the oxidized (Met) species and a second product that is almost certainly superoxide. The second product can be detected by its reduction of cytochrome c3+ at the same rates and in the same amount as the Met species. Anions are increasingly effective as promoters in the same order as their strengths as nucleophiles, e.g. Cl- less than F- less than OCN- less than SCN- less than N3- less than CN-. Rates are directly proportional to anion concentrations. A linear dependence of rate upon [H+] is also observed and can be related to the protonation of a strongly acidic group. Globin from hemoglobin A and hemes with altered 2,4-substituents gave reconstituted hemoglobins that autooxidize at rates that decrease with the electron-withdrawing power of the substituent: acetyl greater than vinyl greater than hydrogen greater than ethyl. Changes in rate with globin structure can be interpreted in terms of steric access to the ligand binding site; the more sterically restricted is the site, the slower is the reaction. The effects of [O2] on the rate vary with the degree of saturation with O2. At high O2 levels (e.g. from saturation to the point where 5% deoxyHbA and 95% oxyHbA are present), a decrease in [O2] results in an increase in the rate of azide-promoted autooxidation. At O2 levels with from 5 to 75% deoxyHbA, the rate remains nearly constant. At still lower levels with oxyHbA less than 25%, the rate decreases as [O2] is lowered. Exposure to CO reduces the rate. The reaction mechanism for anion-induced autooxidation must provide for the stoichiometric formation of a cytochrome c3%-reducing species (presumably, superoxide), the anion acting as a nucleophile in the rate-determining step, the facilitation by protons, the sensitivity of the rate to the electronegativity of heme iron, and the varied effects of [O2] upon the rate. These findings can not be fully accommodated by mechanisms in which the Met species forms either via dissociative loss of superoxide from the oxy species followed by anion binding or via displacement of protonated dioxygen from oxyheme upon nucleophilic attack of the anion at heme iron. A consistent mechanism is the reaction of protonated deoxy species with the anion followed by the reaction of Fe2+-anion complex with O2 to give Fe3+-anion and (formula, see text). Here, the deoxy rather than oxy species is involved in Fe2+ oxidation to Fe3+; O2 can serve as one-electron acceptor but not while serving as an iron-bound ligand. A precise non-iron site for electron transfer from ferrous porphyrin to O2 remains unlocated but a process involving the porphyrin pi-system has analogy in simple heme, flavin, or other organic donor reactions with O2 that yield superoxide... PMID- 6279656 TI - The subunit structures of two distinct receptors for insulin-like growth factors I and II and their relationship to the insulin receptor. PMID- 6279657 TI - The formation of sulfur trioxide radical anion during the prostaglandin hydroperoxidase-catalyzed oxidation of bisulfite (hydrated sulfur dioxide). AB - The mechanism of prostaglandin synthase-dependent (bi)sulfite (hydrated sulfur dioxide) oxidation was investigated using an enzyme preparation derived from ram seminal vesicles. The horseradish peroxidase-catalyzed oxidation of (bi)sulfite was used as a model system. Incubation of (bi)sulfite with prostaglandin synthase and arachidonic acid, 15-hydroperoxyarachidonic acid, or H2O2 results in the formation of the reactive sulfur trioxide anion radical (SO3(-)). The horseradish peroxidase/H2O2 system also oxidizes (bi)sulfite to SO3(-). This free radical reacts with oxygen resulting in oxygen consumption by these incubations. The free radical was detected with the indirect electron spin resonance technique of spin trapping. The SO3(-) radical adduct formed by the reaction of SO3(-) with the spin trap, 5,5-dimethyl-1-pyrroline-N-oxide, gives a nitroxide free radical with a nearly unique electron spin resonance spectrum (aH = 16.0 G and aN = 14.7 G). Using the spin-trapping technique, the SO3(-) could be detected even in incubations of guinea pig lung microsomes. When arachidonic acid-derived prostaglandin G2 was the source of hydroperoxide, formation of SO3(-) could be inhibited by indomethacin. When 15-hydroperoxyarachidonic acid or hydrogen peroxide was used to drive the enzymatic oxidation of (bi)sulfite, indomethacin had no effect. This hydroperoxidase activity was not nearly as heat-labile as the cyclo-oxygenase reaction which forms prostaglandin G2. Finally, the peroxidatic oxidation of (bi)sulfite may occur in vivo in competition with the mitochondrial sulfite oxidase, which oxidizes (bi)sulfite to sulfate without the formation of free radicals. PMID- 6279659 TI - Platelet-derived growth factor. II. Specific binding to cultured cells. AB - We have prepared radioiodinated purified platelet-derived growth factor (125I PDGF) which retains full mitogenic activity. The binding of 125I-PDGF to Swiss 3T3 cells is saturable and highly competed by whole blood serum, purified unlabeled PDGF, and by material from each stage in the purification of PDGF from platelet-rich plasma. Other purified mitogens and substances tested do not compete. 125I-PDGF binding to fibroblasts, 3T3 cells, and arterial smooth muscle cells shows an apparent dissociation constant of 10(-11) M, comparable to the range in which PDGF is mitogenic. A clone of Swiss 3T3 cells obtained from a population selected repeatedly against mitogenic response to PDGF shows a greatly reduced mitogenic response to PDGF and binds only 5% as much 125I-PDGF/cell. The binding capacity of the different cell types tested ranges from 2,500 binding sites/cell on the poorly responding variant to 390,000 binding sites/cell on one strain of Swiss 3T3 cells. Cell types that do not respond to PDGF do not show specific high affinity binding of 125I-PDGF. At 4 degrees C, 125I-PDGF binding to monolayer cultures is relatively slow. Equilibrium binding of low concentrations of 125I-PDGF is not achieved during 7 h unless the binding medium is constantly mixed. 125I-PDGF binding at 4 degrees C shows a broad pH optimum between 6.3 and 8.0. Binding does not seem to require Ca2+ or Mg2+ but is reduced more than 6 fold if both monovalent and divalent salts are omitted. The initial rate of 125I PDGF binding is greater at 37 degrees C than at 4 degrees C but cell-associated 125I begins to decline soon after reaching a peak value at 30-60 min. Coincident with this decline, trichloroacetic acid-soluble 125I appears in the medium and the binding capacity of the cells declines. These phenomena suggest that PDGF and its receptor may be internalized and degraded. PMID- 6279658 TI - Induction of urea cycle enzymes by glucagon and dexamethasone in monolayer cultures of adult rat hepatocytes. PMID- 6279660 TI - Platelet-derived growth factor. III. Identification of a platelet-derived growth factor receptor by affinity labeling. AB - Two homobifunctional cross-linking reagents have been used to cross-link 125I platelet-derived growth factor (PDGF) to a cell surface component with an approximate Mr = 164,000 that has many characteristics of a specific PDGF receptor. Excess unlabeled PDGF competed for labeling of this component, while high concentrations of fibroblast growth factor, insulin, epidermal growth factor, low density lipoprotein or acetylated low density lipoprotein had no effect. Preincubation of cells with 125I-PDGF at 37 degrees C reduced specific 125I-PDGF binding (down regulation) and produced a parallel decrease in the amount of the 164,000-dalton receptor available for labeling. The 164,000-dalton component contains at least some protein that is accessible to trypsin in the extracellular medium. A complex of comparable Mr is seen on all PDGF-responsive cell types examined, but not on a nonresponsive cell type. 125I-PDGF does not become covalently cross-linked to this component in the absence of a cross linking reagent. PMID- 6279661 TI - Bacteriophage lambda DNA packaging in vitro. The involvement of the lambda FI gene product, single-strand DNA, and a novel lambda-directed protein in the packaging reaction. AB - The FI gene product (gp) of bacteriophage lambda is required during phage head assembly in vivo. Mutations in this gene lead to an accumulation of immature concatemeric lambda DNA and of proheads that appear normal and are competent for DNA packaging in vitro. This phenotype can be taken as evidence of a failure to couple DNA and proheads for packaging/maturation. In contrast to the requirement for gpFI in vivo, the packaging of lambda DNA in vitro occurs efficiently in the complete absence of gpFI. However, if ssDNA is included at the outset of the in vitro packaging reaction, DNA packaging is blocked. This block to packaging is relieved by addition of gpFI. Thus packaging of lambda DNA in vitro can be made dependent of gpFI by the inclusion of ssDNA at the outset of the reaction. Inhibition of DNA packaging by ssDNA appears to be mediated by a lambda b region directed protein (packaging inhibitor, ben protein) that is present in the crude extracts of cells used to support the early steps of the packaging reaction. Neither ssDNA nor the packaging inhibitor alone has significant inhibitory effect on packaging; both components are required together to effect the inhibition that is relieved by gpFI. The packaging inhibitor was extensively purified and shown to have endonucleolytic activity. Several lines of evidence are presented to support the idea that both the inhibitory and endonucleolytic activities are functions of the same protein. Although gpFI relieves the inhibition imposed by the ben protein in packaging, gpFI fails to block the DNA cleavage activity of the ben protein in the standard endonuclease assay. PMID- 6279662 TI - A novel endonuclease specified by bacteriophage lambda. Purification and properties of the enzyme. AB - A DNA endonuclease whose expression is under the control of the b region of bacteriophage lambda has been partially purified from an induced lambda lysogen. In a reaction that requires single-stranded DNA, ATP, and Mg2+, the lambda induced endonuclease makes one double strand break in pBR322 and other covalently closed circular DNA molecules, converting these substrates into unit-length linear forms. The double strand break in pBR322 DNA occurs at one of several preferred sites. Linear DNA appears not to be a good substrate for the enzyme. PMID- 6279663 TI - Human transforming growth factor. Production by a melanoma cell line, purification, and initial characterization. AB - A low molecular weight human transforming growth factor (hTGFs) was isolated from serum-free medium conditioned by a human metastatic melanoma tumor line, A2058. The purification of hTGFs was achieved by gel permeation chromatography of the acid-soluble growth-promoting activity on Bio-Gel P-10 in 1 M acetic acid, followed by reverse phase high pressure liquid chromatography on muBondapak C18 support using a linear gradient of acetonitrile in 0.045% trifluoroacetic acid, and subsequently by rechromatography of the human transforming growth factor containing fractions with a linear 1-propanol gradient in 0.035% trifluoroacetic acid on the same column. The estimated molecular weight of hTGFs is 7400. It is a single chain polypeptide with three intrachain disulfide bridges and no free sulfhydryl groups. Lacking tyrosine and methionine, but containing three phenylalanine residues, hTGFs is unlike human and mouse epidermal growth factor (EGF). hTGFs competes with 125I-labeled EGF for binding to A431 human carcinoma cells completely and equivalently, and thus is functionally related to EGF. In contrast, hTGFs enabled normal anchorage-dependent rat kidney cells to grow in soft agar, whereas EGF did not stimulate growth of these cells in semisolid medium. The half-maximal growth-stimulating response of hTGFs was reached with one EGF-competing unit or 1.1 ng of hTGFs. PMID- 6279664 TI - Cytochrome oxidase subunit III gene in Neurospora crassa mitochondria. Location and sequence. AB - We have located and sequenced the gene for cytochrome oxidase subunit III (CoIII) in Neurospora crassa mitochondria. The CoIII gene is located downstream from the small rRNA gene within a cluster of tRNA genes and is coded by the same strand as the tRNA and the rRNA genes. Like the tRNA and the rRNA genes, the CoIII gene is also flanked by the GC-rich palindromic DNA sequences which are highly conserved in N. crassa mitochondria. The CoIII coding sequence predicts a protein 269 amino acids long including 8 tryptophan residues. All 8 tryptophan residues are coded for by UGA. This supports our previous conclusion based on the anticodon sequence of N. crassa mitochondrial tryptophan tRNA and provides evidence for the notion that use of UGA as a codon for tryptophan rather than chain termination may be a feature common to most mitochondrial protein synthesis systems. The close correspondence between the amino acid composition of N. crassa CoIII and that of the protein predicted by the CoIII gene sequence suggests that unlike in mammalian mitochondria, AUA is a codon for isoleucine and not for methionine in N. crassa mitochondria. The N. crassa CoIII sequence shows strong homologies to the corresponding yeast and human proteins (53 and 47%, respectively). The overall hydrophobic character of the protein is consistent with suggestions that most of CoIII is embedded in the mitochondrial inner membrane. PMID- 6279665 TI - Photoaffinity labeling of the beta-adrenergic receptor with azide derivatives of iodoccyanopindolol. AB - Two photosensitive iodocyanopindolol derivatives, 1-(4-azidobenzimidyl)-3,3 dimethyl-6-hydroxy-7-(2-cyano-3-iodoindol-4-yloxy)-1,4-diazaheptane (ICYP-azide 1) and 1-(4-azidobenzoyl)-3,3-dimethyl-6-hydroxy-7-(2-cyano-3-iodoindol-4-yloxy) 1,4-diazaheptane (ICYP-azide-2) have been prepared. [125I]ICYP-azide-1 and -2 (specific radioactivity up to 2.2 Ci/mumol) bind specifically and with very high affinity (KD = 40-45 pM) to beta-adrenergic receptors of turkey erythrocyte membranes. When [125I]ICYP-azide-1 or -2 were incubated with membranes and UV irradiated, two polypeptides (Mr = 40,000 and 50,000) were specifically photolabeled as shown by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. These polypeptides may represent subunits of the beta-adrenergic receptor. The yield of specific covalent label incorporation into both polypeptides was up to 17.2% with [125I]ICYP-azide-2 when expressed as fraction of total beta-receptor binding sites. Since the Mr = 40,000 polypeptide was labeled predominantly and since covalent incorporation had the same concentration dependence as reversible specific binding, this polypeptide could contain a beta adrenergic ligand binding site. Due to the low working concentration (10-100 pM) of [125I]ICYP-azide-1 and -2, nonspecific labeling of membrane proteins was extremely low. The new photoaffinity labels should therefore become valuable tools for probing beta-receptor structure. PMID- 6279666 TI - Sources of variations encountered during the selection and production of three strains of FMD virus for the development of vaccine for use in Nigeria. PMID- 6279667 TI - Mini-review. The postsynaptic alpha 2-adrenoreceptor. AB - The discovery of postsynaptic alpha 2-adrenoreceptors with the drug specificities of presynaptic alpha 2-adrenoreceptors has contributed to a refinement of the classification of alpha-adrenoreceptors. postsynaptic alpha 2-adrenoreceptors have been identified by pharmacological means and with the aid of direct radioligand-receptor binding studies. The evidence for the existence of this class of alpha 2-adrenoreceptors in the brain and in vascular smooth muscle is particularly strong. Central postsynaptic alpha 2-adrenoreceptors play a major part in the hypotensive action of centrally acting antihypertensive drugs such as clonidine and alpha-methyl-DOPA. Vascular smooth muscle cells contain postsynaptic alpha 2-adrenoreceptors which mediate vasoconstriction, like the more classical alpha 1-adrenoreceptors. The simultaneous occurrence of contractile alpha 1- and alpha 2-adrenoreceptors in vascular smooth muscle offers a simple model for the characterization of alpha-adrenoreceptor agonists and antagonists. At present, highly selective agonists of alpha 2-adrenoreceptors have been found. These new compounds may be useful for the classification of other alpha-adrenoreceptor populations. It has been suggested that the vascular postsynaptic alpha 2-adrenoreceptor might be located at extrasynaptic sites. Accordingly, adrenaline released by the adrenal medulla would be the endogenous stimulant. Finally, observations have been made in vivo indicating that a transmembrane influx of calcium ions is necessary for linking the drug-induced activation of these alpha 2-adrenoreceptors to vasoconstriction. PMID- 6279668 TI - Adrenoreceptors in the human penis. AB - 1. Adrenaline, noradrenaline and phenylephrine caused contraction of the corpus cavernosum muscle of the human penis. These sympathomimetic amines did not product inhibitory effects even in the presence of the alpha-adrenoreceptor blocker, phentolamine. The effect of dopamine was similar to that produced by these three sympathomimetic amines. Higher doses of isoprenaline and salbutamol also contracted this preparation. 2. Cocaine and guanethidine enhanced the motor response to adrenaline, noradrenaline and phenylephrine while the action of dopamine was blocked by these two drugs. 3. The motor response to adrenaline, noradrenaline and phenylephrine was antagonized by phentolamine and often potentiated by the beta-adrenoreceptors blocking drug, propranolol. The contractions produced by high doses of salbutamol and isoprenaline were also abolished by phentolamine. These findings indicate that the motor response to sympathomimetic amines is the result of activation of alpha-adrenoreceptors in the corpus cavernosum muscle. 4. At low doses, isoprenaline and salbutamol relaxed the corpus cavernosum muscle strip. The inhibitory action was blocked by low concentrations of the beta-adrenoreceptor antagonist, propranolol but not by practolol (beta 1-adrenoreceptor antagonist) and butoxamine (beta 2 adrenoreceptor antagonist). beta-adrenoreceptors are present in the corpus cavernosum muscle but these are unlikely to be either of the beta 1 or beta 2 type. The possible existence of a third type of beta-adrenoreceptor is suggested. PMID- 6279669 TI - Effect of thyroid status on beta-adrenoreceptors and muscarinic receptors in the rat lung. AB - 1. The effects of thyroid status on the specific binding of the muscarinic ligand (-)-[3H]quinuclidinyl benzilate (QNB) and of the beta-adrenoreceptor ligand (-) [3H]dihydroalprenolol (DHA) in the adult rat lung were investigated. 2. The specific binding of (-)-[3H]quinuclidinyl benzilate (QNB) to lung membranes was saturable and the equilibrium dissociation constant (KD) determined from Scatchard analysis was 54 pM. Kinetic analysis of the binding of [3H]QNB yielded a KD of 42 pM. [3H]QNB binding was inhibited by muscarinic agonists and antagonists, the order of their potency was l-hyoscyamine greater than atropine greater than scopolamine greater than oxotremorine greater than carbachol. These data were consistent with [3H]QNB binding to the muscarine receptor. 3. Adult male rats treated for 2 weeks with the antithyroid agent 3-amino-1,2,4-triazole (ATZ) showed a 52% and 80% reduction in the serum concentration of triiodothyronine (T3) and thyroxine (T4) respectively. These hypothyroid rats also had a 39% decrease in the concentration of lung beta-adrenoreceptors and a 37% decrease in the concentration of lung muscarinic receptors as compared to euthyroid controls. Concurrent treatment of rats with ATZ and T4 for 2 weeks resulted in a reduction of 15% and 20% in the concentration of lung beta adrenoreceptors and muscarinic receptors respectively. The KD values for [3H]DHA and [3H]QNB binding did not change with the ATZ or ATZ+T4 treated groups. 4. Administration of T4 (500 micrograms/kg/day) to male rats for 12 days did not result in any significant change in the concentration of either beta adrenoreceptors or muscarinic receptors compared to euthyroid controls. No change in the KD values for [3H]DHA or [3H]QNB binding were detected. 5. The results show that hypothyroid rats have a reduced lung concentration of both beta adrenoreceptors and muscarinic receptors whereas in hyperthyroid rats these receptors do not significantly change from euthyroid controls. PMID- 6279670 TI - Structure-activity relations for presynaptic inhibition of noradrenergic and cholinergic transmission by adenosine: evidence for action on A1 receptors. AB - 1. This study was designed to determine the structure-activity relations for presynaptic inhibition by adenosine of noradrenergic transmission in rat vas deferens and of cholinergic transmission in guinea-pig ileum. 2. The mid-portion of vasa deferentia and segments of ileum were continuously stimulated at 0.2 Hz and the IC50 values of adenosine analogues for inhibition of the twitch responses determined. 3. In both tissues, loss of the 2' and/or 3' hydroxyl groups resulted in a very marked loss of activity, whereas substitution at the 5' position was compatible with considerable activity. 4. In both tissues, an amino group at C6 on the purine moiety was essential for activity. Halide substitutions at the C2 position and bulky substitutions at the N6 position were compatible with increased activity. 5. The L-diastereomer of N6-phenylisopropyladenosine was approximately 100 times more potent that the D-diastereomer in both tissues. 6. It was concluded that adenosine acts at presynaptic receptors with 'R' site characteristics to inhibit noradrenergic and cholinergic transmission and further that these receptors may belong to the A1 subtype. PMID- 6279671 TI - Stimulation of collagenase secretion from rheumatoid synovial tissue by human collagen peptides. AB - Dayer et al. demonstrated that mononuclear cells from human peripheral blood produce a mononuclear-cell factor that stimulates collagenase secretion from adherent rheumatoid synovial cells. The production of this mononuclear-cell factor can be stimulated by phytohemagglutinin, a T-cell mitogen. We have examined immune mechanisms by which collagenase secretion may be stimulated from explants of rheumatoid synovial tissue and from primary monolayer cultures of rheumatoid synovial cells. Conditioned media from cultures of normal peripheral blood mononuclear cells that had been activated by phytohemagglutinin stimulated collagenase secretion from most explants and from all monolayer cultures that were examined. The direct addition of phytohemagglutinin stimulated collagenase secretion in explants from eight of fourteen patients and in monolayer cultures from three of six patients. These observations indicate the presence of responsive T lymphocytes in rheumatoid synovial tissue and implicate them in the stimulation of collagenase secretion from synovial cells. The direct addition of type-II collagen peptides to rheumatoid explants also stimulated collagenase secretion in explants from six to fifteen patients. Four of five of these patients displayed moderate or severe disease activity. Other patients showed a marginal stimulation of collagenase secretion on addition of type-II collagen peptides (four of fifteen patients) and type-I collagen peptides (three of twelve patients). None of the patients with mild or inactive disease responded to type II collagen. With one exception, the addition of collagen peptides to monolayer cultures did not stimulate collagenase secretion. An increased production of factors that enhance or inhibit the migration of polymorphonuclear leukocytes was also observed in explant cultures in response to the addition of phytohemagglutinin (in seven of twelve patients), of type-I collagen peptides (in five of ten patients), and of type-II collagen peptides (in five of eleven patients). There was no correlation between production of these factors and stimulation of collagenase secretion in rheumatoid synovial tissue. PMID- 6279672 TI - Coccygeal glomus tumors: a case of mistaken identity? AB - We undertook an anatomical and histological study to differentiate glomus-cell tumors of the pericoccygeal tissues from the normal coccygeal body. Removal of the coccyx was performed on five consecutive autopsy specimens from patients with no history of coccygeal symptoms. In each specimen, the coccygeal body (glomus coccygeum) was identified grossly and histologically. The histological appearance was indistinguishable from that of photomicrographs published in case reports of patients with glomus tumors of the coccyx. It is likely that the so-called tumors reported previously were in actuality normal glomus bodies. PMID- 6279673 TI - Expression of a mannosyl-fucosyl receptor for endocytosis on cultured primary macrophages and their hybrids. AB - The presence of a pinocytosis receptor, specific for mannose-fucose terminated glycoproteins, has been established on murine resident peritoneal macrophages, thioglycollate-elicited peritoneal macrophages, and macrophages derived from bone marrow in culture. Macrophagelike cell lines (J-774 and P338.D1), a myelomonocytic cell line (427E), lymphocytes, polymorphonuclear leukocytes, and fibroblasts were negative. Binding and uptake of 125I-mannose-BSA and 125I-beta glucuronidase, respectively, into thioglycollate-induced peritoneal macrophages is saturable (Kd 4 degrees C = 5.4 X 10(-9) M; Kuptake 37 degrees C = 7 X 10(-7) M) and sugar specific. Macrophage-macrophage (rat X mouse) hybrids prepared by fusing rat alveolar macrophages with J-774-B10 (HAT-sensitive macrophagelike cell line) expresses the mannose-fucose receptor. Karyotypes of the hybrids confirmed a 1:1 fusion of rat and mouse cells. The rat/mouse hybrids express a variety of rat and mouse antigens including Fc receptors. Fibroblast-macrophage hybrids and melanoma-macrophage hybrids were negative for mannose-fucose receptor activity. The expression of the mannose-fucose receptor by macrophages appears to be regulated independently of other macrophage markers. PMID- 6279674 TI - Erythroblast cell lines transformed by a temperature-sensitive mutant of avian erythroblastosis virus: a model system to study erythroid differentiation in vitro. AB - A continuous chicken erythroblast cell line transformed by the temperature sensitive mutant ts34 of avian erythroblastosis virus was developed. This cell line, designated HD3, could be induced to terminally differentiate by shift to the nonpermissive temperature. The differentiated cells resembled erythrocytes as judged by morphology, expression of hemoglobin as determined by benzidine staining and radioimmunoassay, and by the expression of differentiation-specific cell surface antigens. Terminal differentiation was dependent on an erythropoietin-like activity present in anemic chicken serum. In contrast, induction of differentiation in the same cells by butyric acid was erythropoietin independent and did not lead to the formation of erythrocytes. In addition, we found that the responsiveness to temperature inducibility and to butyric acid could be dissociated in variant sublines of HD3 and that both types of differentiation inducers appear to act via different pathways. PMID- 6279675 TI - Transforming genes of chicken bursal lymphomas. AB - Bursal lymphomas of chickens infected with lymphoid leukosis viruses contain at least two distinct activated transforming genes: 1) a c-myc gene activated by integration of viral DNA and 2) a cellular gene, not linked to viral DNA, which efficiently induces transformation of NIH 3T3 cells. Since lymphomagenesis by lymphoid leukosis viruses appears to be a multistep process, it is possible that activation of these two different genes occurs at different stages of the disease. The bursal lymphoma transforming gene detected by transfection has been isolated by molecular cloning. Hybridization analysis indicates that this gene is homologous to normal chicken DNA sequences. PMID- 6279676 TI - Insertion of drug resistance genes in animals. PMID- 6279677 TI - SV40-transformed human cells fail to grow in zinc concentrations which permit normal human fibroblast proliferation. AB - Zinc is a metal known to be required for normal growth of both cells, and organisms. When normal and SV40-transformed human tumor cells are plated and grown in medium containing zinc sulfate, a significant fraction of the transformed cells fail to grow at zinc concentrations which are relatively nontoxic to the normal fibroblasts. Although cultured cells respond to certain metallic ions by increasing their metallothionein content, no difference in the ability of normal and transformed cells to produce metallothionein in response to zinc exposure could be detected, thus ruling out this mechanism as a basis for the differing abilities of the cells to grow in zinc in vitro. These results suggest that zinc may be capable of differentially regulating the growth of normal and SV40-transformed human fibroblasts. PMID- 6279678 TI - Membrane transport properties differ following return of serum-deprived versus Ca++-deprived human fibroblasts to a proliferative state. AB - Human lung fibroblasts (W138) can be brought to a quiescent state by removal of serum from the medium or by lowering of the extracellular Ca++. Upon return of Ca++ or serum, the cells enter the G1 phase and progress to S within 15-18 hours. Since multiple G1 phase blocks have been demonstrated, we wished to determine whether the Ca++ and serum block were equivalent since previous data suggested that these two medium components may act at a common point in the initiation of proliferation. We have evaluated the membrane transport of 86Rb, 3-O methylglucose, AIB, and cycloleucine following stimulation of quiescent cells by Ca++ or serum. Serum stimulation results in large increases in the influx of all the substances tested. These increases are prevented if Ca++ is absent upon serum stimulation or they are rapidly diminished following Ca++ removal. In contrast, Ca++ stimulation of Ca++-deprived cells causes little or no enhancement of any of the transport system, yet the cells progress to S phase in a manner similar to serum-stimulated cells. These results indicate that the Ca++ and serum G0 and G1 block are not equivalent and that the serum-induced change in transport of these components does not appear necessary for successful G1 phase progression. Furthermore, the data suggest that the sequence in which Ca++ or serum are presented to the cells alters the ability of Ca++ to modulate the transport systems. Quiescent cells which are exposed to Ca++ prior to serum possess a Ca++ modulation of several transport systems. Cells which are exposed to Ca++ subsequent to serum do not appear to possess this Ca++ regulation. PMID- 6279679 TI - Morphological transformation of cells induced by Kirsten sarcoma virus transforming factor is independent of serine proteases. AB - Reports from several laboratories have suggested that the virus transformed state may be maintained either by ectopically produced growth factors or alternatively by ectopically produced serine proteases including plasminogen activator. Here we show that the maintenance of transformation induced by Kirsten sarcoma virus induced growth factor(s) is independent of serine proteases in that 1) the factors are not themselves serine proteases, and 2) the growth factors do not induce the expression of detectable serine proteases or plasminogen activator. PMID- 6279680 TI - Uptake of [3H]ouabain from the cell surface into the lysosomal compartment of HeLa cells. AB - [3H]Ouabain specifically bound at sublethal concentrations to Na,K-ATPase on the surface of HeLa cells is taken up (internalized) by the cells at a rate of three membrane equivalents of labeled sites per generation. Immediately following a pulse label with the glycoside, codistribution of radioactivity with the surface marker 5'-nucleotidase is found in both conventional sucrose-gradient fractionation and in fractionation following a digitonin treatment. At appropriate concentrations digitonin increases the buoyant density of the HeLa surface membrane and solubilizes the lysosomal marker beta-hexosaminidase (Tulkens et al., 1974). After internalization, [3H]ouabain is also solubilized by digitonin. A shear analysis is described which shows internalized ouabain and beta-hexosaminidase to be codistributed in a particulate fraction that is homogeneous with respect to shear; extrapolation to zero-shear shows that little or none of either marker is found in the soluble fraction of the cytosol. Both markers are coreleased from the particulate fraction by osmotic shock. Although internalized ouabain is subsequently released from these cells with a half-time of about 70 hr, apparently by exocytosis, the shear sensitivity of the remaining cell-associated ouabain does not change for up to 72 hr. Thus ouabain (together with Na,K-ATPase?) appears to be taken up from the surface into a lysosomal compartment and, by at least one criterion, this compartment does not change its physical properties with time, i.e., does not "age." PMID- 6279681 TI - Receptor-associated changes of the catecholamine-sensitive adenylate cyclase in glioma cells doubly transformed with Moloney sarcoma virus. AB - A doubly transformed rat glioma cell line, designated C6V-1, was obtained from rat glioma C6 cells by infection with a rat-adapted variant of Moloney sarcoma virus (MSV-M-os). The C6V-1 cells show karyotypic changes in chromosome number (43) and structure, while C6 cells possess a normal male karyotype. C6V-1 and C6 cells were employed for characterization of a receptor-adenylate cyclase system of the surface membrane. C6V-1 cells showed lower adenylate cyclase activity than that of C6 cells, though the apparent Km for ATP in both types of cells was the same. The maximal stimulation of adenylate cyclase by isoproterenol was significantly reduced, and Kact for isoproterenol was approximately 18-fold lower in C6V-1 cells. When the concentration of beta-adrenergic receptors was measured by various concentrations of [3H] dihydroalprenolol (DHA), the maximal binding sites of C6 and C6V-1 cells were 760 and 230 fmol/mg protein, respectively, without any changes in the association constant for DHA. The concentration of isoproterenol required for 50% displacement of the [3H] DHA binding (Kd) was the same (around 1.5 X 10(-6)M) in both cells, measured in the presence of GTP. Thus the 19-fold drop in the Kd/Kact ratio in C6V-1 cells suggests an incomplete coupling of beta-receptors to adenylate cyclase. Cyclic AMP phosphodiesterase activity and cAMP content in C6V-1 were lower than in C6 cells. Mitochondrial monoamine oxidase and cytosomal enolase activities, however, were somewhat higher in C6V-1 cells. The results indicate that a set of changes in the receptors and in the cyclic AMP system of C6V-1 is one of the specific alterations by transformation, even though those may not be the cause of cell transformation. PMID- 6279682 TI - Mouse embryos contain polypeptide growth factor(s) capable of inducing a reversible neoplastic phenotype in nontransformed cells in culture. AB - A growth-factor-like substance capable of inducing nontransformed mouse AKR-2B, rat NRK, and EGF-receptorless mouse NR6 cells to form progressively growing colonies in soft agar was identified in acid/ethanol extracts of 17-day mouse embryos. This "mouse embryo factor" (MEF) is similar to previously described transforming growth factors in that it is capable of stimulating DNA synthesis and conferring a reversible transformed morphology on nontransformed cells in vitro. Passage of crude embryo extracts over a Bio-Gel P-60 column gave a major peak of soft agar growth-stimulating activity in the 15,000 molecular weight range with a minor peak at about 22,000. This biological activity was sensitive to treatment with either trypsin or dithiothreitol, but was unaffected by heat (56 degrees C for 30 minutes or 100 degrees C for 3 minutes), indicating that the activity is due to a heat-stable polypeptide(s) with disulfide bonds. Separation of these polypeptide(s) by chromatography on carboxymethyl cellulose revealed two peaks of soft agar growth-stimulating activity which did not cochromatograph with a peak of epidermal growth factor receptor-competing activity. The similarities of this mouse embryo-derived growth factor to previously identified transforming growth factors suggest that both fetal development and neoplastic transformation may be affected by similar mechanisms. PMID- 6279683 TI - Regulation of dihydrofolate reductase gene transcription in methotrexate resistant mouse fibroblasts. AB - We have used a methotrexate-resistant mouse 3T6 cell line (M50L3) that overproduces dihydrofolate reductase (DHFR) and its mRNA by a factor of about 300 to study the regulation of DHFR hnRNA synthesis. We have previously shown that when resting (G0) M50L3 cells are serum stimulated to reenter the cell cycle, the amount and rate of synthesis of DHFR and the content of DHFR mRNA all begin to increase as the cells enter the S phase of the cell cycle. The increase in DHFR mRNA content is due to an increase in the rate of mRNA production. In the presnt study, we have used the technique of DNA-excess filter hybridization to determine the rate of synthesis of DHFR hnRNA relative to total hnRNA at various times following serum stimulation. We found that the relative rate of DHFR hnRNA synthesis began to increase at about the same time (6 hours), and increased to about the same extent (three to fourfold by 15 hours following stimulation) as we observed previously for DHFR mRNA production. This suggests that the increase in DHFR mRNA production (and consequently DHFR gene expression) is controlled primarily, if not exclusively, at the level of transcription. We also studied the effect of addition of high concentrations of dibutyryl cAMP and theophylline on DHFR gene transcription. We found that addition of these drugs at the time of stimulation completely blocked the increase in DHFR hnRNA synthesis as well as entry into S phase. Addition of the drugs at either 13 or 20 hours following stimulation led to a rapid decrease in DHFR hnRNA synthesis. The drugs were found to have little effect on the ability of the cells to complete S phase when they were added at 13 hours following stimulation. Our results suggest that high intracellular concentrations of cAMP may affect DHFR gene expression not only by preventing the progression of cells through the G1 phase of the cell cycle but also by affecting the rate of DHFR gene transcription in a more direct manner. PMID- 6279684 TI - Separation of glycopeptides by high performance liquid chromatography. AB - A method is presented for separation of tryptic glycopeptides-containing oligosaccharides of the N-asparagine-linked type. High performance liquid chromatography (HPLC) of glycopeptides on a C18 reverse-phase system eluted with a gradient of 0%-50% acetonitrile in 0.1 M NaPO4 pH 2.2 resolves the two major glycosylation sites from the envelope glycoprotein (G) of vesicular stomatitis virus. Glycopeptides containing N-linked oligosaccharides of the complex type coelute with those containing N-linked oligosaccharides of the neutral, high mannose type, indicating that separation is based upon peptide rather than carbohydrate composition. The contribution of the carbohydrate component to glycopeptide elution, as determined by cleavage of the high mannose oligosaccharides with endo-beta-Nacetylglucosaminidase H, is that of a significant, but minor, decrease in peptide retention time. Comparison of the tryptic glycopeptide profiles of G isolated from both wild type and mutant strains of VSV illustrates the rapid, reproducible, and quantitative nature of the technique. Through HPLC analysis of appropriately treated glycopeptides, it is possible to explore both the nature and extent of glycosylation at individual sites in glycoproteins in a single step. PMID- 6279685 TI - The phosphomannosyl recognition system for intracellular and intercellular transport of lysosomal enzymes. PMID- 6279686 TI - Metabolism of benzo(a)pyrene by human epithelial and fibroblastic cells: metabolite patterns and DNA adduct formation. AB - We demonstrate in cell culture that mammary epithelial cells from normal human breast specimens metabolize benzo(a)pyrene (BaP) and form adducts with the bases of their DNA more readily and at lower concentrations of BaP than do fibroblasts from the same specimens. BaP metabolism and adduct formation was determined in the same incubations with epithelial cells grown out in early passage from each of three specimens and with fibroblasts from one of these specimens. The metabolite pattern of the epithelial cells was indicative of preferential formation of 7, 8-dihydrodiol-9, 10-dihydroepoxybenzo(a)pyrene the ultimate carcinogen. In contrast, fibroblasts formed mainly mono- and dihydroxide derivatives of BaP. The metabolite pattern from epithelial cells was compatible with the ease in which adducts between DNA and the diolepoxide of benzo(a)pyrene were formed. These results provide evidence that chemical carcinogens should be considered as possible factors in the induction of breast cancer in women. PMID- 6279687 TI - Carcinogen-mediated amplification of specific DNA sequences. AB - A model experimental system based on SV40-transformed Chinese hamster embryo cells and a highly sensitive in situ hybridization procedure was designed. Exposure of the cells to different categories of chemical and physical carcinogens resulted in the induction of SV40 DNA synthesis in the treated cells. Although the carcinogen-mediated amplification of SV40 DNA sequences is regulated by the viral "A" gene, neither infectious virus nor complete viral DNA molecules were rescued from the treated cells. A heterogenous collection of DNA molecules containing SV40 sequences was generated following treatment with DMBA. Restriction enzyme analysis of the amplified DNA molecules in the Hirt supernatant revealed that not all sequences in the integrated SV40 inserts are present. The possibility that the amplification of SV40 sequences is a reflection of a general gene amplification phenomenon mediated by carcinogens is discussed. PMID- 6279688 TI - Chemotactic receptors of Dictyostelium discoideum. PMID- 6279689 TI - Enzyme-linked immunosorbent assay for detection of antibody to varicella-zoster virus. AB - Primary varicella-zoster virus (VZV) infection is a serious illness in immunocompromised individuals, and a rapid, sensitive, and reliable assay to identify high-risk VZV-susceptible patients would be clinically useful. An enzyme linked immunosorbent assay (ELISA) for antibody to VZV was compared with the fluorescent antibody-to-membrane antigen (FAMA) assay and found to be similar in both sensitivity and specificity. The antibody titers determined by both assays were also similar. The absence of antibody detected by ELISA correlated with susceptibility to VZV infection. Because it is simple to perform and has equivalent sensitivity to FAMA, ELISA should be useful for VZV antibody testing in diagnostic and research laboratories. PMID- 6279690 TI - Influence of humidity on rotavirus prevalence among Nigerian infants and young children with gastroenteritis. AB - Rotaviruses were detected by counterimmunoelectro-osmophoresis in the feces of 16 (13.8%) of 116 infants and young children with gastroenteritis during a 5-month period (September 1979 through January 1980) in Ife, Nigeria. The rate of rotavirus detection varied inversely with relative humidity and was highest in December (38.5% positive) when the humidity was lower. There was not such a distinct relationship with temperature or vapor pressure; and although the rate of rotavirus detection was higher in the drier months (November to January; 19.3% positive) than in the rainy season months (September and October; 8.5% positive), the average humidity was lower in the drier months than in the rainy months. Low relative humidity (49 to 78%) is apparently the most important environmental factor for rotavirus survival and spread in this area. PMID- 6279691 TI - Improved immunofluorescence antigens for detection of immunoglobulin M antibodies to Epstein-Barr viral capsid antigen and antibodies to Epstein-Barr virus nuclear antigen. AB - Epstein-Barr viral capsid antigen and nuclear antigen produced by modified procedures were evaluated for use in measuring viral capsid antigen immunoglobulin M and Epstein-Barr virus nuclear antigen antibody responses in sera from patients with suspected Epstein-Barr virus infections. Viral capsid antigen production was stimulated with a phorbol ester, and the Epstein-Barr virus nuclear antigen cells were fixed in suspension to eliminate loss of antigen during the drying process. Both preparations proved to be sensitive and reliable. PMID- 6279692 TI - Typing of herpes simplex virus with type-specific human immunoglobulin M in an indirect immunofluorescence assay. AB - Sera from nine individuals with suspected primary herpes simplex virus type 1 (HSV-1) or type 2 (HSV-2) infection were screened to identify those containing HSV type-specific immunoglobulin M (IgM). Selected sera were then utilized in an IgM-specific indirect immunofluorescent-antibody HSV-typing assay (patent pending). To evaluate the procedure, 29 HSV isolates were grown in cultures of continuous human amnion cells, fixed, and used as substrates for indirect immunofluorescence. Determination of virus type was based on intensity of fluorescence of the substrate with HSV-1- and HSV-2-specific antisera after staining with fluorescein-conjugated anti-human IgM. Typing of the isolates by restriction endonuclease digestion showed that of 29, 18 were HSV-2 and 11 were HSV-1. Results by IgM-specific indirect immunofluorescent-antibody assay were identical to those by restriction endonuclease digestion for 27 of the isolates; 2 isolates failed to replicate adequately in the test cells. The IgM-specific indirect immunofluorescent-antibody procedure appears to be a simple, rapid, and accurate technique which could be of use to clinical virology laboratories. PMID- 6279693 TI - Porcine pararotavirus: detection, differentiation from rotavirus, and pathogenesis in gnotobiotic pigs. AB - Some characteristics of a newly recognized porcine enteric virus are described. Tentatively, the virus was referred to as porcine pararotavirus (PaRV) because it resembled rotaviruses in respect to size, morphology, and tropism for villous enterocytes of the small intestine. However, it was antigenically distinct from porcine, human, and bovine rotaviruses and reoviruses 1, 2, and 3, and the electrophoretic migration pattern of PaRV double-stranded RNA was distinct from the electrophoretic migration patterns of the rotaviral and reoviral genomes. By passage in gnotobiotic pigs, PaRV was isolated from two suckling diarrheic pigs originating from two herds. After oral exposure of gnotobiotic pigs, villous enterocytes of the small intestines became infected as judged by immunofluorescence, resulting in villous atrophy and diarrhea. Mortality was high when gnotobiotic pigs less than 5 days old were infected. The C strain of this virus was serially passed 10 times in gnotobiotic pigs, and electron microscopy, immunofluorescence, and serological tests indicated no extraneous agents. The virus was serially passed five times in cell cultures which contained pancreatin in the medium, but replication was negligible or absent, as the number of immunofluorescent cells decreased with each passage. Since rotaviral infections are frequently diagnosed by direct electron microscopy of fecal specimens, the presence of other morphologically similar viruses, such as PaRV, should be considered. The use of immune electron microscopy is suggested as a means of helping recognize this situation. PMID- 6279694 TI - Comparison of indirect fluorescent-antibody amoebic serology with counterimmunoelectrophoresis and indirect hemagglutination amoebic serologies. AB - Patients ranged from those with no prior diagnosis of or suspected exposure to Entamoeba histolytica to those with proven amoebic liver abscesses (extraintestinal disease). A comparison of serologies from patients with proven and suspected amoebiasis or possible past exposure revealed good correlation between the indirect fluorescent antibody (IFA) procedure and the other methods used, counterimmunoelectrophoresis and indirect hemagglutination. Titers from patients with proven extraintestinal amoebiasis were in the expected high range previously reported by other authors. Patients with clinical histories suggestive of exposure to E. histolytica but no proven disease had lower titers which indicated possible background exposure. The IFA procedure provides a rapid method of antibody detection; results obtained on an emergency basis provide essential information in making the diagnosis of amoebic abscess, pyogenic abscess, or tumor. The IFA procedure is rapid, reliable reproducible, and relatively inexpensive to perform, provided a good source of antigen is consistently available. PMID- 6279696 TI - Differentiation of herpes simplex virus types 1 and 2 by sensitivity to (E)-5-(2 bromovinyl)-2'-deoxyuridine. AB - Selective inhibition of herpes simplex virus (HSV) type 1 replication but not of HSV type 2 replication by (E)-5-(2-bromovinyl)2'-deoxyuridine (BVDU) was utilized to differentiate clinical isolates of these viruses. BVDU (0.7 microgram/ml) reduced HSV type 1 PFU by an average of 2.74 log10, but had no effect on the plaque-forming ability of HSV type 2 isolates. The incorporation of BVDU into the media of primary cell cultures used for virus isolation in the clinical laboratory allowed for the correct presumptive identification of HSV types. PMID- 6279695 TI - Detection of infectious mononucleosis heterophil antibody by a rapid, standardized enzyme-linked immunosorbent assay procedure. AB - A rapid, specific, sensitive, standardized, a reproducible enzyme-linked immunosorbent assay (ELISA) procedure has been developed for detecting the heterophil antibody associated with infectious mononucleosis (IM). The IM heterophil antibody used for the solid phase was purified from bovine erythrocyte stroma. The test uses heavy-chain-specific anti-immunoglobulin M (IgM) labeled with alkaline phosphatase and three 10-min incubations. The quantitative results correlated well with horse erythrocyte agglutination titers. Absorption tests confirmed the specificity of the ELISA reactions for IM heterophil antibodies. Neither very high levels of IgM in myeloma sera nor high levels of rheumatoid factor caused false-positive reactions. A number of probable IM cases were encountered which positive by ELISA but negative by the horse erythrocyte slide agglutination test. Absorption studies indicated that these were true-positives for the IM heterophil antibody. The IM heterophil antibodies were confirmed to be predominantly of the IgM class, but moderate proportions of the IgM class were sometimes encountered. PMID- 6279697 TI - Identification of DNA viruses by membrane filter hybridization. AB - The use of membrane filter hybridization for the identification of DNA viruses is described. We designed and used a procedure for identification of herpes simplex virus. This method can discriminate between herpes simplex virus types 1 and 2 in a simple way. PMID- 6279698 TI - An antiidiotypic antibody that recognizes the beta-adrenergic receptor. AB - Antialprenolol rabbit antibodies were fractionated on an acebutolol affinity resin, followed by L-propranolol elution so as to separate a class of binding sites that mimic the beta-adrenergic receptor. Allotype-identicaL rabbits were immunized with this fraction. After 6 mo, antisera exhibited antiidiotypic activity inhibiting [3H]alprenolol binding to the original antibody and to rabbit antiacebutolol antibodies, which had a spectrum of ligand-binding properties identical to the original idiotype. Those antisera demonstrating the original idiotype. Those antisera demonstrating the most potent antiidiotypic activity also blocked [3H]alprenolol binding to the beta-adrenergic receptor of turkey membrane, canine pulmonary membrane, and rat reticulocyte. An idiotype affinity purified fraction showed similar activity, inhibiting beta-receptor binding with a calculated dissociation constant (KD) of 53 nM. Isoproterenol-mediated adenylate cyclase activity was also inhibited in a competitive manner. The universality of recognition of these antiidiotypic antisera indicate that the three-dimensional structure of a receptor's binding site can be modeled by a subset of an elicited antibody population. PMID- 6279699 TI - Glucocorticoid and mineralocorticoid effects on adrenocorticotropin and beta endorphin in the adrenalectomized rat. AB - Immunoreactive ACTH (ir-ACTH) and immunoreactive beta-endorphin (ir-betaEP) were determined in plasma, anterior pituitary, neuro-intermediate lobe, and hypothalamus of sham-adrenalectomized rats, and adrenalectomized rats given six daily injections of vehicle (oil), dexamethasone, 9alpha-fluorocortisol or deoxycorticosterone. 6 d after adrenalectomy, anterior pituitary ir-ACTH and ir betaEP were double, and plasma levels approximately fivefold those in controls. Adrenalectomy did not alter hypothalamic levels of either peptide, or ir-betaEP in neuro-intermediate lobe, in which tissue ir-ACTH was below detection limit at routine dilutions. Dexamethasone (0.2-200 mug/d) concurrently suppressed plasma ir-ACTH and ir-betaEP, with a near maximal effect at 20 mug, and a half-maximal effect between 2 and 6 mug; similar dose-response characteristics were found for thymolysis. Step-wise increases in anterior pituitary content of both peptides were found, with no change in hypothalamic levels of either peptide, or neuro intermediate lobe ir-betaEP. 9alpha-fluorocortisol (0.2-200 mug/d) produced plasma, anterior pituitary, and hypothalamic effects equivalent to dexamethasone, but with one-tenth the potency. Unlike dexamethasone, higher doses of 9alpha fluorocortisol significantly elevated neuro-intermediate lobe ir-betaEP. Deoxycorticosterone (2-2,000 mug/d) produced no significant changes in plasma, anterior pituitary or hypothalamic levels of either peptide; like 9alpha fluorocortisol, doses of >60 mug/d significantly elevated neuro-intermediate lobe ir-betaEP. Whereas ir-ACTH and ir-betaEP synthesis in and release from the anterior pituitary are under complex negative feedback glucocorticoid control, there exists a mineralocorticoid-specific effect on neuro-intermediate lobe content of ir-betaEP. PMID- 6279700 TI - Lysosomal localisation of parallel tubular arrays in chronic lymphocytic leukaemia of T cell origin: an ultrastructural cytochemical study. AB - An ultrastructural cytochemical study of lysosomal acid phosphatase was performed on leukemic cells in a case of chronic lymphocytic leukaemia of T cell origin (T CLL). The cells showed inclusion bodies known as parallel tubular arrays, which often lay within acid phosphatase-positive, membrane-bound spaces. This suggests their lysosomal location. PMID- 6279701 TI - Indomethacin and prostaglandins: their behavioral relationships in an acute toxic psychosis. PMID- 6279703 TI - A study of camelpox in Somalia. PMID- 6279702 TI - Effects of single dose ACTH 4-10 versus placebo on the memory of symptomatic geriatric volunteers. PMID- 6279704 TI - Metastasizing pancreatic islet cell tumors in the rat. PMID- 6279706 TI - The syndrome of milker's nodules in burn injury: evidence for indirect viral transmission. AB - Four patients with first- to second-degree burns developed multiple unusual nodular lesions confined to the burned areas 2 to 3 weeks after the accident. Electron microscopy disclosed viral particles within epidermal cells. These were identified as subgroup II poxvirus. Viral culture established the diagnosis of paravaccinia (milker's nodule) infection. Since none of the patients had had direct contact with infected animals, but had been in contact with contaminated objects, an indirect viral transmission, previously not reported for milker's nodules, appears the most likely mode of infection. PMID- 6279705 TI - The pathology of liver tumours in the dog. PMID- 6279707 TI - Reactive and neoplastic epithelial alterations of the vulva. A classification of the vulvar dystrophies from the dermatopathologist's viewpoint. AB - The clinical classification of the vulvar dystrophies has traditionally been difficult because most of these lesions present as a white hyperkeratotic patch. An international committee for the study of the vulva designed a histologic classification that consists of: I, hyperplastic type, with or without atypia; II, lichen sclerosus; and, III, mixed dystrophy, with or without atypia. A new classification and a fourth type are proposed by us to sort out a group of strictly benign dermatoses with distinctive histopathologic features which can present clinically as a white patch in the vulva; these are best designated as benign dermatoses. In addition to these, squamous cell carcinoma in situ and Paget's disease may also be manifested as a white patch. Hence, a classification of vulvar diseases with new nomenclature is presented from the dermatopathologist's perspective, based on histologic and clinical features. PMID- 6279708 TI - Acyclovir comes of age. PMID- 6279709 TI - Retinoid-binding proteins. AB - Much has been learned during the past decade about the specific retinoid-binding proteins that exist in plasma and in the intracellular compartment in a number of tissues. Vitamin A is mobilized from liver stores and transported in plasma in the form of the lipid alcohol retinol, bound to a specific transport protein, retinol-binding protein (RBP). A great deal is now known about the chemical structure, metabolism, and biologic roles of RBP. A number of tissues of rats, humans, and other species contain soluble binding proteins with specificity for either retinol (cellular retinol-binding protein, CRBP) or retinoic acid (cellular retinoic acid-binding protein, CRABP). These intracellular proteins have been purified from several tissues and partly characterized. This paper provides a concise review of our current knowledge about the three retinoid binding proteins, RBP, CRBP, and CRABP. PMID- 6279710 TI - Effects of all-trans-retinoic acid (retinoic acid) and 4-hydroxyphenylretinamide on synovial cells and articular cartilage. AB - We studied the effects of two retinoids, naturally occurring all-trans-retinoic acid (retinoic acid) and the synthetic 4-hydroxyphenylretinamide (4-OH-PRT) on monolayer cultures of rabbit synovial fibroblasts and on explants of rabbit articular cartilage. Treatment of fibroblasts with phorbol myristate acetate (PMA; 10(-8) M) induced the synthesis and secretion of large amounts of collagenase: this was inhibited if the cells were treated with retinoic acid (10( 6) M) or dexamethasone (10(-7 M). Combined treatment with retinoic acid and the steroid prednisolone, at concentrations as low as 19(-10) M, gave an additive inhibition of collagenase production. Both retinoids inhibited collagenase production, but only 4-OH-PRT prevented the increase in prostaglandin E2 (PGE2) induced by PMA. Levels of plasminogen activator were also increased by treatment with PMA, and concomitant addition of either retinoid further enhanced this stimulation. Possible toxicity was assessed by measuring release of glycosaminoglycans (GAG) from explants of articular cartilage. Treatment with retinoic acid induced release of 80% of the total GAG, whereas treatment with 4 OH-PRT resulted in release of 40% of the total, a finding similar to that seen with untreated samples. 4-OH-PRT inhibited production of collagenase and PGE2 by rabbit synovial fibroblasts but was not toxic to articular cartilage. PMID- 6279711 TI - Inhibition of collagen degradative enzymes by retinoic acid in vitro. AB - The effects of a variety of retinoids on collagenase and gelatinase expression have been examined in skin fibroblast cultures derived from normal volunteers and from patients with the hereditary blistering disorder, recessive dystrophic epidermolysis bullosa. Both 13-cis- and all-trans-retinoic acid were effective inhibitors of collagenase production in both cell types. In the case of collagenase, the inhibition of collagenase activity was paralleled by a reduction in immunoreactive enzyme protein, suggesting that these retinoids act by inhibiting synthesis and/or secretion of the enzyme. Retinoic acid also inhibited production of the second enzyme in the collagen degradative pathway, gelatinase. In this case, the decrease in gelatinase activity was equal to or slightly greater than the achieved in collagenase expression. The observation that certain retinoids modulate the two crucial enzymes in the degradation of collagen in the skin suggests that they might be useful therapeutic agents in recessive dystrophic epidermolysis bullosa, a disease in which the pathogenesis of blistering is in part related to connective tissue destruction. PMID- 6279712 TI - The effects of retinoids on neutrophil functions in vitro. AB - Vitamin A (retinol) and its analogues (retinoids) are clinically effective in cystic acne and psoriasis, diseases in which neutrophils may constitute major components of inflammatory cell infiltrates. We found that the earliest histopathologic alteration in psoriasis is the disappearance of neutrophils at 2 to 4 weeks after the initiation of therapy with etretinate. Since retinoids may exert anti-inflammatory effects by virtue of an action upon neutrophils, we studied the effects of the following retinoids on discrete neutrophil functions in vitro: retinol, retinyl acetate, retinal, tretinoin, isotretinoin, etretinate, and Ro 10-1670. Although they had no significant effects upon aggregation, chemokinesis, or chemotaxis, all of the retinoids, with the exception of etretinate and Ro 10-1670, profoundly inhibited superoxide anion production and lysosomal enzyme release. Tretinoin and isotretinoin were the most effective inhibitors. We propose that these drugs exert their pharmacologic effects (resolution of inflammatory lesions) by inhibiting the release of mediators of inflammation and by preventing the accumulation of neutrophils in acne lesions when applied topically or systemically, respectively. PMID- 6279714 TI - Radiographic blood pool contrast agents for vascular and tumor imaging with projection radiography and computed tomography. AB - An emulsion of Ethiodol and Pluronic was evaluated as a vascular (blood pool) imaging agent with both the GE experimental scanned projection radiography system and conventional computed tomography (CT), 475 mg iodine/kg body weight of emulsion, was injected into 11 rabbits with thigh muscle V2 carcinomas. Both projection radiography scans and CT scans visualized aorta, vena cava, pulmonary, and femoral vessels, and distorted vessels around tumors for 20 min after infusion. This persistent vascular visualization allowed CT scanning at multiple contiguous levels, which could be followed by reformatting in the sagittal and coronal planes. Following diatrizoate in identical iodine/kg doses, vascular visualization persisted for only 1 min after infusion. Blood pool agents are ideal for projection radiography systems and CT reformatting of vascular anatomy. PMID- 6279713 TI - Drug states as discriminative stimuli in a flavor-aversion learning experiment. AB - Injection of poison into rats after they drank in the presence of stimulus compounds of a drug state and a flavor resulted in little stimulus control by the drug state. In Experiment 1, half of the rats were poisoned after drinking salt water while stimulated with amphetamine and after drinking sugar water while sedated with pentobarbital, but they were not poisoned after salt-pentobarbital or sugar-amphetamine combinations. The other half were subjected to counterbalanced procedures. In abstract language, poisoning occurred after AX and BY stimulus combinations but did not occur after AY and BX combinations. In Experiment 2A, rats were poisoned only after consuming a particular flavored solution (salt or vinegar) in a particular state (pentobarbital or undrugged); that is, if AX was poisoned, BX, BY, and AY were experienced without poisoning. There was complete counterbalancing of flavors and drug states. Experiment 2B was similar except that amphetamine was used instead of pentobarbital. In both experiments, there was some discrimination learning based on the drug state, gut it was extremely weak. PMID- 6279715 TI - Mesoblastic nephroma. PMID- 6279717 TI - Update on drug therapy: VIII. Oral cephalosporins. PMID- 6279716 TI - Distant, late metastases to skin of carcinoma of the breast. AB - Three cases of late, distant metastases to the skin of cancer of the breast and a review of the literature are presented. PMID- 6279718 TI - American Medical Women's Association, Inc., proposed Constitution and Bylaws changes. PMID- 6279719 TI - Preparation of Na,K-ATPase-containing liposomes with predictable transport properties by a procedure relating the Na,K-transport capacity to the ATPase activity. AB - A microprocedure for the preparation of Na,K-ATPase-containing liposomes with a minimal starting material (200 microgram) of purified Na,K-ATPase is presented. Phosphatidylcholine is added gradually to cholate-solubilized Na,K-ATPase of various concentrations and the lipid-induced decrease in enzyme activity is monitored. After removal of the detergent by dialysis, the transport parameters of the resulting Na,K-ATPase-liposomes are established by a microassay. By relating the transport properties to the Na,K-ATPase activity preset before dialysis, a procedure is developed which allows to prepare standardized Na,K ATPase-liposomes with predictable transport properties. PMID- 6279720 TI - Evidence of degradation of polybrominated biphenyls in soil samples from Michigan. AB - Soil samples obtained from the former polybrominated biphenyls (PBB) manufacturing site in Michigan were analyzed by gas chromatography and gas chromatography with mass spectrometric detection. The results indicate significant degradation of the PBB residue in the soil sample. The soil sample with the highest concentration of PBB had the greatest degree of degradation. Principal degradation products include 2,3', 4,4', 5-pentabromobiphenyl, 2,2', 4,4', 5-pentabromobiphenyl and two unidentified tetrabromobiphenyls. The degradation pattern observed supports a photochemical decomposition mechanism. These degraded residues may be more toxic than the original Firemaster residues. The implications of the results are discussed. PMID- 6279721 TI - [The neurophysiology of micturition (author's transl)]. AB - The ortho and parasympathetic nervous system as well as the somatic nervous system make up the innervation of the lower urinary tract. This constitutes the autonomic nervous system of these organs. There are control centres in the cortex, the sub-cortex and the medullary areas of the brain as well as this peripheral innervation. Together they make possible by their various connections a variety of regulatory circuits and a series of reflexes for micturition which bring about a complex assembly coming from the cortex to the sphincter mechanism of the bladder. PMID- 6279722 TI - [Phosphonate esterase activity in blood serum during early pregnancy (author's transl)]. PMID- 6279723 TI - Gelatin degrading activity in early and term human placenta. PMID- 6279724 TI - Glomus tumor associated with pacinian hyperplasia--case report. AB - Features of the glomus tumor and the relatively recently described pacinian hyperplasia are described including signs and symptoms, differential diagnosis and treatment. The role of trauma as an etiology is proposed. A case is reported and discussed in which the patient had a glomus tumor and pacinian hyperplasia in an index finger, and a glomus tumor in the small finger of one hand. Treatment of both entities was surgical excision. PMID- 6279725 TI - Anterior submuscular transposition of the ulnar nerves by the Learmonth technique. AB - Thirty-eight patients with progressive posttraumatic ulnar neuropathy at the elbow underwent anterior submuscular transposition of their nerves. Multiple mechanical causes of neuropathy were identified. Fourteen patients had undergone previous surgery for ulnar neuropathy, while 24 had not. Postsurgical follow-up averaged 23.1 months. The operative technique is described and illustrated in detail. Complications attributable to surgery were minimal. No absolute prognostic factors could be identified, and even those patients with significant muscular atrophy or time delay before operation were generally benefited. If prior surgery had induced significant scarring and neural damage, the prognosis for recovery was considerably worse, as it also was for patients who had severe preoperative dysesthesia or pain. Four patients thought to represent examples of double crush or compression syndrome were identified. PMID- 6279726 TI - An unusual presentation of a ring injury. AB - A case of chronic erosion of the soft tissues and bone of the proximal phalanx of the ring finger in a 62-year-old woman is presented. The patient's rings had eroded the skin, nerves, and tendons of the finger as well as the palmar aspect of the proximal phalanx; however, circulation was adequate. When the rings were removed, there was no improvement in sensibility or range of motion, but this was not bothersome to the patient. PMID- 6279727 TI - A high-fiber diet does not cause mineral and nutrient deficiencies. AB - We evaluated the hypothesis that a healthy population taking a high-fiber diet may develop deficiencies of various minerals and nutrients in three groups of healthy subjects: 1) sixty-eight people regularly supplementing their diet with at least 2 tablespoons of bran for at least 6 months (mean 3 tablespoons for 13 months); 2) forty-three controls not consuming bran supplements; and 3) twenty vegetarians (eight of them consuming bran supplements) who had a very high fiber consumption for many years. The mean serum levels of iron, total iron binding capacity, calcium, phosphorus, alkaline phosphatase, zinc, magnesium, vitamin A, and cholesterol were within the normal range in all the three groups. In the bran eaters, vitamin A was higher, and alkaline phosphatase and magnesium lower than in the other two groups. In the vegetarians, the phosphorus level was higher and cholesterol and iron binding capacity lower than in the other two groups. There was no correlation between the amount of bran consumed and the blood level of nutrients. The fiber consumption of the vegetarians was very high, more than three times that of the controls. Our study indicates that a moderately or even extremely high consumption of fiber for a long time does not by itself cause mineral or nutrient deficiencies in a western type population. PMID- 6279729 TI - Should we eat more fibre? AB - Controversy continues over the role of dietary fibre in health, and whether or not a standard fibre intake should be recommended for the UK. In this review an attempt is made to consider the evidence for and against making such a recommendation. In doing so, it is clear that many problems exist in studies examining the role of fibre in disease, such as the inherent inadequacies of epidemiological studies and distrust of such studies by some circles, or the lack of uniformity in definition of dietary fibre and in its chemical analysis. The effect of fibre on faecal bulking is given as an example of experimental verification of epidemiological findings which has led to the widespread use of fibre in treating diverticular disease and constipation. Evidence of beneficial effects for cancer of the colon and ischaemic heart disease are far less convincing. Few harmful effects of fibre have been documented, apart from continuing disagreement regarding fibre and mineral balance, a question which remains to be solved. On weighing the evidence, it is suggested that recommending higher-fibre intakes in the UK is a favourable guideline. The type of fibre to be recommended is discussed, based on new evidence of the mode of action of different types of fibre in the gastrointestinal tract, and the amount of fibre considered, in relation to intakes in other countries and in the past in the UK. PMID- 6279728 TI - Transposable elements controlling genetic instabilities in mammals. AB - It is proposed that the instabilities in gene action of some alleles at certain loci in the mouse (e.g., a, c, H-2 Mi, p, pe, T, Va, W), which do not seem to conform to traditional hypotheses of gene action, are better interpretable in the light of modern studies of transposable DNA elements (insertion sequences and transposons of prokaryotic organisms; controlling elements of maize; transposable controlling elements of Drosophila). Some phenotypic evidence in the mouse in support of this hypothesis is presented for the a, Mi, p, and W loci, which affect pigmentation. PMID- 6279730 TI - Effect of cellulose on serum lipids in adolescent girls. AB - The effect on serum lipid concentration of adding 21 g cellulose to a controlled low-fibre diet of natural foods was studied in nine healthy adolescent girls. The experimental period consisted of two periods of three weeks with each subject serving as her own control. The addition of cellulose did not affect serum total lipids, cholesterol, phospholipids, triglycerides and free fatty acids. PMID- 6279731 TI - A two-step procedure for obtaining normal peripheral blood T-lymphocytes using continuous equilibrium density gradient centrifugation on percoll. AB - Equilibrium centrifugation of either peripheral blood mononuclear cells or of pure lymphocytes (obtained by carbonyl iron or glass bead adherence removal of monocytes) on a continuous density gradient of Percoll yielded lymphocyte fractions containing between 92 and 99% T lymphocytes as shown by sheep red blood cell rosetting. B lymphocytes with surface immunoglobulin were found in the regions of low density (1.03-1.065 g/ml) and T lymphocytes in the regions of higher density (1.06-1.08 g/ml). TM lymphocytes with their characteristic positive 'dot' pattern of staining for non-specific esterase were also found mainly in regions of high density. It was concluded that Percoll continuous equilibrium density gradient centrifugation can be used to obtain T lymphocytes in high yield, with high viability and without metabolic changes which may occur after contact with sheep red blood cells. The esterase staining suggested that there was also some separation of T lymphocyte subsets. PMID- 6279733 TI - Decreased accumulation of cyclic adenosine 3',5'-monophosphate in "ischemic" skin after 12-0-tetradecanoyl-phorbol-13-acetate treatment. AB - The effect of 12-0-tetradecanoyl-phorbol-13-acetate (TPA) on cyclic adenosine 3',5'-monophosphate (cyclic AMP) level in adult mouse skin in response to ischemia was examined. The incubation of skin pieces in a buffered salts medium at 37 degrees C resulted in a rapid accumulation of cyclic AMP. In mouse skin pieces maximum accumulation (about 6 times the basal level) occurred after 2 min incubation and was followed by a rapid decline in the cyclic AMP level. This "ischemic" rise in epidermal cyclic AMP was greatly reduced if skin was used 16 hr after a single application of 17 nmoles of TPA. The effect of TPA on cyclic AMP accumulation in response to ischemia was first observed at 1 hr after TPA treatment and was maximal at 4 hr. The lack of "ischemic" response in TPA-treated skin was not related to an increase in the activity of cyclic AMP phosphodiesterase after TPA application. In addition, the accumulation of cyclic AMP in skin in response to both ischemia and exposure to isoproterenol, adenosine, histamine, or prostaglandin E2 (PGE2) was not observed in skin treated with the tumor promoter TPA. PMID- 6279732 TI - A role for anti-inflammatory agents and cyclic AMP in regulating fibronectin mediated phagocytosis. AB - The present study deals with the effects of anti-inflammatory drugs and agents known to elevate intracellular levels of cyclic AMP (cAMP) on plasma fibronectin mediated (PFn) phagocytosis of radiolabeled, gelatin-coated latex particles (g Ltx*) by inflammatory macrophages. Monolayers of casein-elicited peritoneal macrophages were preincubated with the specified agents for either 1 or 24 hrs at 37 degrees C prior to the measurements of phagocytosis in the presence of human plasma fibronectin (47 microgram/ml) and heparin (6.7 U/ml). Under these conditions, prostaglandin E1, colchicine, vincristine, and cytochalasin B were all effective in inhibiting g-Ltx* phagocytosis by macrophages in a dose dependent fashion. More potent inhibition of phagocytosis was manifested by agents known to increase intracellular levels of cAMP in phagocytic cells. Dibutyryl cyclic AMP (dbcAMP), d,1-isoproterenol and aminophylline (10(-5) to 10( 3) M) were all effective in reducing the uptake of g-Ltx* by macrophages. The combination of dbcAMP and aminophylline acted additively. These studies demonstrate that anti-inflammatory drugs and cAMP-elevating agents exert potent inhibitory effects on fibronectin-mediated phagocytosis of gelatin-coated particles by macrophages. Thus, our system provides a suitable in vitro model for further investigations into the humoral regulation of phagocytosis of denatured collagen-coated particles and tissue debris by inflammatory phagocytic cells. PMID- 6279734 TI - Acquisition of serum antibody to Norwalk Virus and rotavirus and relation to diarrhea in a longitudinal study of young children in rural Bangladesh. AB - Serum antibodies to Norwalk virus and to rotavirus were measured during longitudinal studies of infectious diseases and nutrition in rural Bangladesh. Initially, the prevalence of antibody to Norwalk virus was 7% in children younger than six months and increased to 80% in children two to five years of age. The incidence of titer increases was highest in one- and two-year-olds and in children who had low or undetectable levels of antibody. Some Norwalk virus infections appeared to result in diarrhea. Nearly all children had serum antibodies to rotavirus at the beginning of the study; however, children with the lowest levels of antibody to rotavirus had the greatest risk of rotavirus diarrhea. Over half of the children had a fourfold increase in titer of antibody to rotavirus during the year, and 7% had increases in two of the three study periods during the year. Most increases in titer of antibody to rotavirus appeared to result from subclinical infections. PMID- 6279735 TI - In vivo effects of enterotoxin from Clostridium perfringens type A in the rabbit colon: binding vs. biologic activity. AB - Rabbit colonic loops were exposed to purified enterotoxin from Clostridium perfringens type A and perfused for analysis of the effects of the enterotoxin on transport and surface structure of the colon. The colon showed no apparent response to the enterotoxin in the transport of fluid and electrolytes as compared with untreated animals. There was a significant increase in levels of luminal fluid protein, corresponding to an observed increase in secretion of mucus. Histologic sections showed no epithelial damage, but scanning electron microscopy showed an increase in the number of mucous fields over the value in control animals. The colon showed little or no overt response to enterotoxin treatment, but isolated colonic epithelial cells bound 125I-labeled enterotoxin at levels even greater than those observed in cells from the ileum, where the enterotoxin has considerable biologic activity. Thus, although binding of enterotoxin is necessary for its biologic activity, biologic activity does not necessarily follow binding to specific receptors. PMID- 6279736 TI - Enhanced susceptibility of mice infected with murine cytomegalovirus to intranasal challenge with Escherichia coli: pathogenesis and altered inflammatory response. AB - Three-week-old mice inoculated intraperitoneally with murine cytomegalovirus (MCMV) and then challenged intranasally with Escherichia coli strain K1 demonstrated enhanced mortality (70%-90%) as compared with control animals infected with either pathogen alone (0-20%, P less than 0.05). Mortality was greatest when animals were challenged with E. coli on days 1 or 3 after MCMV inoculation. On day 3 of infection with MCMV, clearance of E. coli from blood and tissues was impaired, and there was a decreased inflammatory response to an E. coli-inoculated sponge implanted subcutaneously (geometric mean of 830 leukocytes in sponge fluid/mm3 in MCMV-infected animals vs. 8,510 leukocytes/mm3 in controls, P less than 0.01). On days 1 and 3 of MCMV infection, decreased leukocyte counts in sponge fluid correlated with increased levels of bacteremia (P less than 0.05). These results indicate that MCMV enhances susceptibility to an intranasal challenge with E. coli. A decrease in the inflammatory response may be one mechanism by which MCMV increases susceptibility to bacterial infections. PMID- 6279737 TI - Seroepidemiology of Epstein-Barr virus in pregnant women. AB - Sera from 2,000 women in the early stages of pregnancy in 1980-1981 were tested for antibodies to Epstein-Barr virus. Of the 2,000 women, 1,386 attended an obstetrical clinic (average age, 21 years) and 614 attended a private practice (average age, 28 years). Only eight (0.6%) of the women attending the obstetrical clinic were seronegative (less than or equal to 20 years, five [0.8%] of 650; 21 30 years, three [0.5%] of 648; and greater than or equal to 31 years, zero of 88), whereas 18 (2.9%) of the women attending the private practice were seronegative (less than or equal to 20 years, two [4.8%] of 42; 21-30 years, 12 [3.0%] of 388; and greater than or equal to 31 years, four [2.3%] of 184). Probable recent seroconversions were detected in 35 women. Sera from 185 women pregnant in 1959-1965 were tested for antibodies to Epstein-Barr virus; 17 (9%) were seronegative, compared with 26 (1.3%) of the women in 1980-1981 (P less than 0.01). PMID- 6279738 TI - Therapeutic effects of foscarnet sodium and acyclovir on cutaneous infections due to herpes simplex virus type 1 in guinea pigs. AB - The therapeutic effects of topically applied foscarnet sodium and acyclovir on cutaneous infections due to herpes simplex virus type 1 (HSV-1) were studied in guinea pigs. Foscarnet cream (3%) reduced both the cumulative vesicle score and the time for healing when HSV-1 strains C42 and 79 were tested. The application of foscarnet in cream form resulted in a dose-dependent reduction in skin virus titers and vesicle scores at concentrations ranging from 0.05% to 3%. Infections due to HSV-1 strain 79, but not those due to strain C42, were sensitive to treatment with 5% acyclovir in polyethylene glycol ointment. Topical application of 3% acyclovir in dimethyl sulfoxide on infections due to HSV-1 strain C42 resulted in a reduction in the skin virus titers, lesion scores, and the time for healing. Acyclovir, in both polyethylene glycol and dimethyl sulfoxide, was consistently less active than foscarnet cream in these animal experiments. PMID- 6279739 TI - Disseminated infection due to Mycobacterium avium-intracellulare in a homosexual man with Kaposi's sarcoma. PMID- 6279740 TI - [A case of adenoid cystic carcinoma of the trachea treated by segmental resection and primary anastomosis of the trachea (author's transl)]. PMID- 6279741 TI - [Close classification method of undetermined cases of trophoblastic disease (author's transl)]. AB - With the improvement of recovery ratio of trophoblastic disease by means of primary chemotherapy, undetermined cases of trophoblastic diseases, whose morphological examination has not been conducted, have increased. The respective sickly constitutions of these cases are extremely complicated, and we have deviced a new close classification of these cases from clinical standpoint. For the purpose, we have chosen out the following items.: A. Major subdivision 1. Types of antecident pregnancy. 2. Duration between the antecident pregnancy and the commencement of chemotherapy. 3. Urinary hCG secretion pattern following molar pregnancy. 4. Following treatment of hydatidiform mole. B. Minor subdivision 1. Urinary hCG titer at the commencement of chemotherapy. 2. BBT pattern. 3. Chest x-ray findings. 4. PAG. 5. Clinical signs. 6. Development of focuses (clinical). 7. Necessary reasons for chemotherapy. These items are expressed respectively by figures. This close classification method is based on the combinations of these figures. The authors believe, this method will enable the easy classification of early chemotherapy examples and will present a strong key for the analysis of enormous cases. PMID- 6279742 TI - [Studies on influence of synthetic LH-RH and dbcAMP on normal chorionic villi and BeWo cells (author's transl)]. AB - The influence of synthetic LH-RH and dbcAMP on human chorionic villi and BeWo cells was studied through measurement of cAMP, hCG and estradiol. The results obtained were as follow: 1) There was a rapid increase of cAMP in chorionic villi after the 7th week of gestation, marking a maximum value at the 9th week, and gradually decreasing thereafter. 2) When dbcAMP was added to chorionic villi, there was a significant increase estradiol production in chorionic villi and hCG secretion in media. 3) The stimulation of cAMP in chorionic villi and hCG secretion into medial resulting from the addition of synthetic LH-RH to chorionic villi showed significant increases at 5 minutes and 30 minutes, respectively. 4) When dbcAMP or synthetic LH-RH was added to BeWo cells, increased secretion of hCG and estradiol into media was seen. It therefore is concluded that there is a relationship between the activity of synthetic LH-RH in which cAMP participates and a mechanism involving hCG secretion and estradiol production originating from human chorionic villi and BeWo cells. This is turn suggest the possibility of the existence in chorionic villi of a LH-RH like substance which plays a role in the production or secretion of hormone in the placenta. PMID- 6279743 TI - [Management of retained secundines by hysteroscopy (author's transl)]. AB - Seventy-three patients with retained products of conception were selected from a series of 112 women examined by hysteroscopy, to assess abnormalities of pregnancy. Thirty of the 73 were diagnosed as cases of retained secundines and treated effectively, under hysteroscopic control. This approach has proved to be an effective and a safe procedure for the management of retained secundines. Our study also suggested that traditional therapeutic and/or diagnostic dilatation and curettage (D & C) should be re-assessed and replaced with more effective, simple and safe procedures, such as suction curettage. In these patients with other abnormalities of pregnancy, such as ectopic pregnancy and trophoblastic disease, hysteroscopy does not supplant other diagnostic procedures: rather, it complements them. PMID- 6279744 TI - [Plasma ACTH, beta-lipotropin and beta-endorphin in the human fetus and their mother during delivery (author's transl)]. AB - Adrenocorticotropic hormone (ACTH), beta-lipotropin (beta-LPH) immunoreactivity (IR) and beta-endorphin (beta-EP) immunoreactivity (IR) were measured by high sensitive radioimmunoassay in maternal and umbilical cord plasma samples which were obtained simultaneously in 12 cases. Mean IR-beta-LPH in maternal (690.1 +/- 138.6 pg/ml, +/- S.E.) and cord venous plasma (1114.8 +/- 94.2 pg/ml) were significantly higher than that of normal adults (93.0 +/- 8.2 pg/ml). Mean IR beta -EP in maternal (125.8 +/- 24.0 pg/ml) and cord venous plasma (130.6 +/- 20.6 pg/ml) were significantly higher than that of normal adults (6.0 +/- 0.9 pg/ml). There were significant positive correlations between IR-beta-LPH and IR beta-EP in maternal and cord venous plasma. Mean beta-EP to beta-LPH molar ratio of 0.19 +/- 0.03 in maternal plasma was significantly (p less than 0.05) higher than that of cord venous plasma (0.13 +/- 0.01). There was no significant correlation between the levels of these three hormones in maternal plasma and the levels in cord venous plasma. These data suggest that ACTH, IR-beta-LPH and IR beta-EP were elevate during labor, delivery responding the stress. Beta-LPH and beta-EP in cord venous plasma were fetal and/or placental origin. PMID- 6279745 TI - [The study for the standard of the remission of the patients with trophoblastic disease (author's transl)]. AB - The recurrence of the patients who had obtained remissions once was studied and the standard of remission was reassessed. The comparison of the titers of hCG ranged the LH levels was studied by means of some methods of measurement. 1) During 7 years, from 1973-1979, 141 patients with trophoblastic disease in Chiba University Hospital were treated. The remission rate was 92.2% (130/141), the recurrence rate was 6.2% (8/130). In the high risk group, the recurrence rate was 23.1% (6/26), in the low risk, it was 1.9% (2/104). Four of 8 with recurrence were died, 4 (2 high risk and 2 low risk) obtained remissions again. 2) The mean HCG titers of patients with recurrence were 29.5 +/- 23.5 IU/L (HAR), 13.7 +/- 6.4 mIU/ml (HCG-RIA), 1.5 +/- 0.7 ng/ml (HCG-beta RIA) and that of patients with remission were 13.8 +/- 13.0 IU/L, 11.3 +/- 4.1 mIU/ml, 1.0 +/- 0.6 ng/ml, respectively. 3) The 2 patients whose HCG titers indicated all negative followed the recurrences. The HCG beta titers of the patients who were extirpated their bilateral ovaries indicated rather high in spite of getting their remissions. 4) The essential standard of remission was the equanimity of the HCG titers around the negative levels without disturbance by LH or other substances for a while. But it is not enough for a remission. PMID- 6279746 TI - Hypersensitivity reactions to adriamycin: two case reports. PMID- 6279747 TI - Occupational asthma. PMID- 6279748 TI - Impaired glucagon-stimulated glucose output in livers of acutely uremic rats. AB - The role of glucagon in the pathogenesis of abnormalities of glucose metabolism associated with renal failure remains undefined. We have evaluated glucagon stimulated glucose and cyclic AMP output and amino acid uptake in isolated perfused livers of rats with experimentally-induced ARF and sham-operated controls. ARF animals exhibited azotemia, hyperglycemia, hyperinsulinemia, and hyperglucagonemia. During stimulation with physiologic (3 X 10-10M) or supraphysiologic (3 X 10-8M) glucagon concentrations, glucose output was lower in livers of ARF rats than in those of controls, whereas cyclic AMP responses were similar or exceeded those of controls. Hepatic glycogen content was lower in rats with ARF and the stores were exhausted at the end of perfusions. Additional studies in livers of fasted animals revealed no significant differences in glucose output or amino acid uptake between ARF and control livers perfused with physiologic levels of glucagon. These experiments suggest that the decreased glucagon-stimulated glucose output in isolated perfused livers in acutely uremic rats is due primarily to glycogen depletion rather than to impaired gluconeogenesis. Normal or increased cyclic AMP responses to glucagon suggests intactness of the hormone receptor-adenylate cyclase. PMID- 6279749 TI - Dual system of intestinal thiamine transport in humans. AB - The transport of thiamine across the intestine has been characterized in rats but has not been adequately studied in humans. To determine the kinetics of thiamine intestinal transport directly in humans, mucosal tissues were obtained during routine endoscopy from normal-appearing sites at the second portion of the duodenum. With 3H-dextran as the marker of adherent volume, the uptake of 14C thiamine hydrochloride by the excised mucosa was measured in vitro. By this method thiamine uptake was linear with tissue weight and with incubation time up to 5 min. Results showed that at low thiamine concentrations (0.2 to 2.0 microM), uptake was saturable whereas at high concentrations (5 to 50 microM), uptake was linear with thiamine concentrations. Pyrithiamine, anoxia, N-ethylmaleimide, and replacement of sodium chloride by mannitol reduced the uptake of 0.5 microM thiamine by 42%, 37%, 32% and 35%, respectively (p less than 0.05) but had no effect on the uptake of 20 microM thiamine. These data suggest that, as in the rat, the intestinal transport of thiamine in humans proceeds by a coexistent dual system. At physiologic concentrations, thiamine is transported primarily by an energy-requiring, sodium-dependent active process, whereas at higher pharmacologic concentrations thiamine uptake is predominantly a passive process. PMID- 6279750 TI - Accuracy of localization of acute myocardial infarction by 12 lead electrocardiography. AB - Until recently, ECG accuracy in localizing acute myocardial infarction (AMI) could be assessed only by comparing the ECGs with autopsy findings. This approach, however, preselected patients, including only those who died. It is possible that this postmortem group of patients would be different from the whole population of patients with AMI. Myocardial imaging with 99mTc-pyrophosphate offers the advantage of directly localizing the region of injured myocardium in the acute phase of AMI. In 34 patients with confirmed AMI and focal uptake of 99mTc-pyrophosphate, serial ECGs were obtained and interpreted by two independent observers. The sensitivity and specificity of serial ECGs in determining the location of AMI in the five left ventricular (LV) wall segments were determined: (1) in the anterior wall sensitivity was 86.7% and specificity was 89.5%; (2) in the lateral wall sensitivity was 73.7% and specificity was 80.0%; (3) in the high lateral wall sensitivity was 80.0% and specificity was 87.5%; (4) in the inferior wall sensitivity was 87.5% and specificity was 100%; (5) in the "true" posterior wall sensitivity was 83.3% and specificity was 86.4%. Overall, in the 170 LV wall segments (five per patient) examined, scans localized with a sensitivity of 81.9% and a specificity of 88.8%. After four patients with LBBB were excluded, sensitivity increased to 87.1%. Overall, localization of AMI by serial ECG was accurate in 85.9% of the 34 patients included in the study. PMID- 6279751 TI - Negative rate-sensitive feedback effects on adrenocorticotrophin secretion by cortisol in normal subjects. AB - Plasma ACTH and corticosteroid levels were measured in normal subjects during constant infusion of either 0.9% (W/V) NaCl solution or cortisol, and during insulin-induced hypoglycaemia. During infusions of 0.9% NACl solution the secretion of ACTH and corticosteroids was episodic. Fast, rate-sensitive, negative feedback inhibition of ACTH secretin was observed during cortisol infusions, when the corticosteroid levels were within the physiological range (200-750 nmol/l) and were rising at a rate of between 5 and 10 nmol/l per min for 30 min or longer. When plasma corticosteroid levels were in a steady state, the initial fast feedback effects were abolished and ACTH secretion resumed. However, this recovery of ACTH secretion was not seen when the corticosteroid levels were persistently above 800 nmol/l. It appears that corticosteroid-induced negative feedback in man may be both rate- and level-sensitive. During insulin stress test ACTH secretion fell at time when the plasma corticosteroid level was rising rapidly (greater than 5 nmol/1 per min) despite persistent hypoglycaemia. PMID- 6279752 TI - Effect of human plasma on the bioactivity of adrenocorticotrophin. AB - The effect of plasma on the bioactivity of ACTH was investigated with the cytochemical section bioassay. Levels of ACTH in normal human plasma were underestimated if the plasma concentration in the incubation medium exceeded 2% and if the plasma ACTH level was less than 2 ng/l. A retrospective analysis of 250 plasma ACTH assays revealed that the ACTH concentration was consistently underestimated in the 1:100 dilution compared with the 1:1000 dilution, although not to the extent that any assay would be rejected for non-parallelism. The extent of the underestimation increased as the ACTH concentration increased. Guinea-pig adrenal sections responded faster to 1:10 or 1:50 dilutions of human plasma (containing 2 ng ACTH/l) than to either 1:100 or 1:1000 dilutions of that same plasma, or to ACTH (over the range 0.005-5 ng/l)in the absence of plasma. This faster response could also be seen in the absence of plasma by increasing the ACTH concentration above 5 ng/l. It seems that plasma contains factors which potentiate the biological effect of ACTH in the cytochemical assay system.U PMID- 6279753 TI - Modulation by catecholamines and thyrotrophin of cyclic AMP response to beta adrenergic stimulation by cultured porcine thyroid cells. AB - Thyroid cell cyclic AMP synthesis is stimulated by beta-adrenergic agonists. We have characterized this sensitivity on cultured porcine thyroid cells and have studied its modulation by chronic treatment with thyrotrophin. The synthesis of cyclic AMP in intact porcine thyroid cells in primary culture was stimulated by the beta-adrenergic agonist, isoproterenol. This stimulation was dose-dependent and was inhibited by the beta-adrenergic antagonists propranolol and alprenolol. The cell responsiveness (i.e. the response elicited by 5 microM-isoproterenol after 5-min stimulation) was increased when the cells were cultured in the absence of thyrotrophin. Thyrotrophin, when present in the culture medium at the onset of culturing, inhibited this increase. A concentration of 100 microunits. thyrotrophin/ml was sufficient to reduce the cyclic AMP response to 15% of its control value. Prostaglandin E2 or dibutyryl cyclic AMP did not mimic the effect of thyrotrophin. The low sensitivity of thyrotrophin-treated cells to beta adrenergic agonists could be explained by a decreased number of beta-adrenergic receptors. [125I]Iodohydroxybenzyl pindolol specific binding was ten times greater in membrane preparations of control cells than in membranes derived from thyrotrophin-treated cells. The beta-adrenergic sensitivity of cultured thyroid cells was also decreased after long-term treatment by terbutaline. A time- and dose-dependent desensitization was observed. PMID- 6279754 TI - Effect of high-dose oestrogen administration on the growth and prolactin receptor content of N-nitrosomethylurea-induced mammary tumours in the rat. AB - The effect of chronic administration of a high dose of oestradiol benzoate (OB) on growth and prolactin receptor content of the N-nitrosomethylurea (NMU)-induced rat mammary tumour was investigated. Daily i.m. administration of 50 microgram OB for 8 days to tumour-bearing rats induced marked reduction in tumour size which was already significant after only three injections. Simultaneous daily administration of 1 mg perphenazine to stimulate endogenous prolactin release counteracted the inhibitory effect on tumour growth of OB but only partially since the tumours grew significantly more when perphenazine was injected alone. No treatment significantly affected prolactin receptor content in the tumours. We concluded that administration of pharmacological doses of OB has a potent antitumour action on the NMU-induced rat mammary tumour. This effect can only partially be explained by inhibition of the peripheral action of prolactin. PMID- 6279756 TI - Conduction and block by organic cations in a K+-selective channel from sarcoplasmic reticulum incorporated into planar phospholipid bilayers. AB - A collection of organic cations has been used to probe the gross structural features of the ionic diffusion pathway in a K+-selective channel from sarcoplasmic reticulum (SR). Channels were incorporated into planar phospholipid bilayer membranes, and single-channel currents were measured in the presence of ammonium-derived cations in the aqueous phases. Small monovalent organic cations are able to permeate the channel: the channel conductance drops sharply for cations having molecular cross sections larger than 18-20 A2. Impermeant or poorly permeant cations such as tetraethylammonium, choline, and glucosamine, among others, block K+ conduction through the channel. This block is voltage dependent and can be described by a one-site, one-ion blocking scheme. 19 monovalent organic cations blocks primarily from the trans side of the membrane (the side defined as zero voltage), and much more weakly, if at all, from the cis side (to which SR vesicles are added). These blockers all appear to interact with a site located at 63% (average value) of the electric potential drop measured from the trans side. Furthermore, block by 1,3-bis[tris(hydroxymethyl) methylamino] propane (BTP) shows that the presence of a blocking ion increases the duration of the apparent open state, as expected for a scheme in which the blocking site can be reached only when the channel is open. The results lead to a picture of the channel containing a wide (at least 50 A2) nonselective trans entry in series with a narrow (20 A2) constriction. PMID- 6279757 TI - Kinetics of intramembrane charge movement and sodium current in frog node of Ranvier. AB - Intramembrane charge movement (Q) and sodium current (INa) were monitored in isolated voltage-clamped frog nodes of Ranvier, ON charge movements (QON) for pulses from the holding potential (-100 mV) to potentials V less than or equal to 0 mV followed single exponential time courses, whereas two exponentials were found for pulses to V greater than or equal to 20 mV. The voltage dependence of both QON and its time constant tauON indicated that the two ON components resolved at V greater than or equal to 20 mV were also present, though not resolvable, for pulses to V less than or equal to 0 mV. OFF charge movements (QOFF) monitored at various potentials were well described by single exponentials. When QOFF was monitored at -30 or -40 mV after a 200-microsecond pulse to +20 mV and QON was monitored at the same potential using pulses directly from -100 mV, tauON/tauOFF = 2.5 +/- 0.3. At a set OFF potential (-90 to -70 mV), tauOFF first increased with increasing duration tON of the preceding pulse to a given potential (0 to +30 mV) and then decreased with further increases in tON. The declining phase of tauOFF followed a time course similar to that of the decline in QOFF with tON. For the same pulse protocol, the OFF time constant tauNa for INA also first increased with tON but then remained constant over the tON interval during which tauOFF and QOFF were declining. After 200- or 300 microsecond pulses to +20, +20, or +50 mV, tauOFF/tauNa at -70 to -90 mV was 1.2 +/- 0.1. Similar tauOFF/tauNa ratios were predicted by channel models having three identical charged gating particles that can rapidly and reversibly form an immobile dimer or trimer after independently crossing the membrane from their OFF to their ON locations. PMID- 6279755 TI - Effects of adrenocortical and gonadal steroids on the secretion in vitro of corticotrophin and its hypothalamic releasing factor. AB - The effects of adrenocortical and gonadal steroids on the secretion in vitro of ACTH by adenohypophysial segments and corticotrophin releasing factor (CRF) by isolated hypothalami were studied in the rat. Corticosterone (1.25 X 10(-6) mol/l), betamethasone (2.5 X 10(-8) mol/l) and progesterone (2.5 X 10(-7) mol/l) reduced the hypothalamic extract-induced secretion of ACTH by pituitary tissue in vitro but aldosterone (2 X 10(-7) mol/l), testosterone, androsterone, androstenedione (2 x 10(-7) mol/l), oestradiol, oestriol, and oestrone (10(-6) mol/l) did not. Corticosterone (2.5 +/- 10(-9) mol/l), aldosterone (2 X 10(-8) mol/l) and betamethasone (2 X 10(-10) mol/l) inhibited and oestradiol, oestriol and oestrone (10(-8) - 10(-6) mol/l) potentiated the production of CRF by isolated hypothalami which occurred when acetylcholine or 5-hydroxytryptamine were added to the incubation medium but progesterone (2.5 X 10(-7) mol/l), testosterone, androsterone and androstenedione (2 X 10(-7) mol/l) had no effects. The results indicate that hypothalamo-pituitary-adrenocorticotrophic activity may be modified not only by glucocorticoids but also by other steroids. PMID- 6279760 TI - Simultaneous assay of dihydroxyacetone synthase and transketolase in a methylotrophic yeast grown in continuous culture. A cautionary note. AB - The methylotrophic yeast Candida boidinii CBS 5777 was grown in continuous culture under carbon limitation on glucose, glucose plus methanol, and methanol as carbon and energy sources. During adaptation from glucose to methanol there was a rapid rise in the specific activities of triokinase, fructose-1,6 bisphosphatase and dihydroxyacetone synthase, which are key enzymes of the xylulose phosphate cycle of formaldehyde fixation. The specific activity of classical transketolase fell during this adaptation. Extracts from carbon-limited C. boidinii contained an enzyme which catalysed oxidation of NADH when some preparations or ribose 5-phosphate were added, which was not a transketolase. This enzyme activity was dependent on an impurity in such ribose 5-phosphate preparations and can be confused with transketolase activity. PMID- 6279758 TI - Calcium accumulation by the sarcoplasmic reticulum in two populations of chemically skinned human muscle fibers. Effects of calcium and cyclic AMP. AB - In previous efforts to characterize sarcoplasmic reticulum function in human muscles, it has not been possible to distinguish the relative contributions of fast-twitch and slow-twitch fibers. In this study, we have used light scattering and 45Ca to monitor Ca accumulation by the sarcoplasmic reticulum of isolated, chemically skinned human muscle fibers in the presence and absence of oxalate. Oxalate (5 mM) increased the capacity for Ca accumulation by a factor of 35 and made it possible to assess both rate of Ca uptake and relative sarcoplasmic reticulum volume in individual fibers. At a fixed ionized Ca concentration, the rate and maximal capacity (an index of sarcoplasmic reticulum volume) both varied over a wide range, but fibers fell into two distinct groups (fast and slow). Between the two groups, there was a 2- to 2.5-fold difference in oxalate supported Ca uptake rates, but no difference in average sarcoplasmic reticulum volumes. Intrinsic differences in sarcoplasmic reticulum function (Vmax, K0.5, and n) were sought to account for the distinction between fast and slow groups. In both groups, rate of Ca accumulation increased sigmoidally as [Ca++] was increased from 0.1 to 1 microM. Apparent affinities for Ca++ (K0.5) were similar in the two groups, but slow fibers had a lower Vmax and larger n values. Slow fibers also differed from fast fibers in responding with enhanced Ca uptake upon addition of cyclic AMP (10(-6) M, alone or with protein kinase). Acceleration by cyclic AMP was adequate to account for adrenaline-induced increases in relaxation rates previously observed in human muscles containing mixtures in fast-twitch and slow-twitch fibers. PMID- 6279759 TI - Calcium and cyclic GMP regulation of light-sensitive protein phosphorylation in frog photoreceptor membranes. AB - In frog photoreceptor membranes, light induces a dephosphorylation of two small proteins and a phosphorylation of rhodopsin. The level of phosphorylation of the two small proteins is influenced by cyclic GMP. Measurement of their phosphorylation as a function of cyclic GMP concentration shows fivefold stimulation as cyclic GMP is increased from 10(-5) to 10(-3) M. This includes the concentration range over which light activation of a cyclic GMP phosphodiesterase causes cyclic GMP levels to fall in vivo. Cyclic AMP does not affect the phosphorylations. Calcium ions inhibit the phosphorylation reactions. Calcium inhibits the cyclic GMP-stimulated phosphorylation of the small proteins as its concentration is increased from 10(-6) to 10(-3) M, with maximal inhibition of 70% being observed. Rhodopsin phosphorylation is not stimulated by cyclic nucleotides, but is inhibited by calcium, with 50% inhibition being observed as the Ca++ concentration is increased from 10(-9) to 10(-3) M. A nucleotide binding site appears to regulate rhodopsin phosphorylation. Several properties of the rhodopsin phosphorylation suggest that it does not play a role in a rapid ATP dependent regulation of the cyclic GMP pathway. Calcium inhibition of protein phosphorylation is a distinctive feature of this system, and it is suggested that Ca++ regulation of protein phosphorylation plays a role in the visual adaptation process. Furthermore, the data provide support for the idea that calcium and cyclic GMP pathways interact in regulating the light-sensitive conductance. PMID- 6279761 TI - A conjugative 'plasmid' lacking autonomous replication. AB - Attempts were made to isolate open and covalently closed circular DNA from strains containing the IncJ plasmids. All of the methods tried were unsuccessful. It was shown that the IncJ plasmid R391 can integrate into the Escherichia coli K12 chromosome and can mobilize chromosomal markers from a single origin in an orientated manner. It is proposed that the IncJ plasmids are integrated in the chromosome for most, if not all, of their existence and this explains the inability to isolate plasmid DNA from strains containing them. PMID- 6279762 TI - Is the IS1-flanked r-determinant of the R plasmid NR1 a transposon? AB - The 23 kilobase multiple drug resistance r-determinant (r-det) of the R plasmid NR1 is an IS1-mediated transposon, Tn2671. Drug-resistant Escherichia coli transductants isolated after infection with bacteriophage P1::Tn2671 derivatives carry the intact r-det in their chromosomes. Independently isolated transductants carry the r-det at different locations on the chromosome. From the E. coli chromosome, Tn2671 can transpose to various locations on the phage P7 genome. Throughout these processes, r-det is maintained as a stable unit. Various possible molecular mechanisms, which all might contribute with characteristic frequencies to the transposition of Tn2671, are discussed. The results presented are relevant to the understanding of mechanisms for a wide spreading of drug resistance genes. PMID- 6279763 TI - Oxolinic acid-resistant mutants of Bacillus subtilis. AB - Two mutants of Bacillus subtilis have been independently selected for resistance to oxolinic acid, and inhibitor of DNA gyrase. The mutations, designated oxr-1 and oxr-2, map very close to one another but are clearly separated from mutations in the genes for DNA gyrase. Many of the phenotypic properties of the mutants differ from those of a strain containing the gyrA mutation described by other workers. In particular, the oxr strains are as sensitive as the wild-type to inhibition by nalidixic acid on solid medium. In addition, experiments with DNA synthesis in toluenized cells show that the enzyme of the gyrA mutant is resistant to oxolinic acid, whereas DNA synthesis in the oxr mutants is as sensitive as it is in wild-type preparations. It is concluded that resistance to oxolinic acid is not due to an alteration in the DNA gyrase, but is more probably the result of an impaired uptake of the inhibitor. Although growth of the mutants on agar plates is inhibited at high concentrations of oxolinic acid, lower concentrations (1-2 microgram ml-1) can be used to distinguish them from the wild type. The oxr-1 and oxr-2 mutations define a new genetic locus and can be used as genetic markers in B. subtilis. PMID- 6279764 TI - Inhibition of sporulation by DNA gyrase inhibitors. AB - The effects of oxolinic acid and novobiocin - which respectively inhibit the A and B subunits of DNA gyrase, and therefore inhibit DNA synthesis - have examined in sporulating cultures of Bacillus subtilis. Although both inhibitors prevent sporulation, this is not due to inhibition of DNA synthesis. Instead, they affect protein synthesis generally, though weakly, but have very marked effects on the formation of individual enzymes. These effects are reproducible, but the type of enzyme that will be affected is not predictable. The results point to an involvement of DNA gyrase in the transcription of some genes. This is suggested as the reason for the effect of the inhibitors on spore formation, which they block mainly at Stage O-I. PMID- 6279765 TI - Characterization of conjugative R plasmids belonging to the new incompatibility group IncU. AB - Five conjugative plasmids governing different antibiotic resistance patterns were identified in wild strains of enteric bacteria isolated in Czechoslovakia and the G.D.R. between 1976 and 1979. They have been characterized as members of the new incompatibility group IncU (reference plasmid RA3 from Japan). The molecular sizes of the IncU plasmids ranged between 18 and 37 megadaltons; their restriction fragment patterns indicated them to be distinct types. PMID- 6279766 TI - Regulation of extracellular alkaline protease activity by histidine in a collagenolytic Vibrio alginolyticus strain. AB - Vibrio alginolyticus synthesized an inducible extracellular collagenase in a peptone medium during the stationary growth phase. These cultures also possessed extracellular alkaline serine protease activity. The alkaline protease activity did not require a specific inducer and it was produced in tryptone or minimal media. The collagenase was not produced in either the tryptone or minimal media. The alkaline protease activity was sensitive to catabolite repression by a number of carbon sources, including glucose, and by amino acids and ammonium ions. Cyclic AMP, dibutyryl cyclic AMP and cyclic GMP did not relieve catabolite repression. Histidine and urocanic acid stimulated the production of alkaline protease activity in tryptone and minimal media. Other compounds associated with the histidine utilization (hut) pathway did not increase alkaline protease activity. Histidine reversed the repression of alkaline protease activity by glucose of (NH4)2SO4 in minimal medium. Histidine and the compounds associated with the hut pathway inhibited collagenase production. PMID- 6279767 TI - Relationship between sporulation medium and germination ability of Mucor racemosus sporangiospores. AB - Asexual sporangiospores of Mucor racemosus produced on a minimal sporulation medium (M spores) germinated only if glucose, mannose or a complex substrate such as peptone, yeast extract or Casamino acids was present. Once germinated, growth was supported by a wide range of substrates including amino acids, carbohydrates or organic acids. Sporangiospores produced on a nutritionally complex sporulation medium (C-spores) germinated on a wide range of carbon sources. C-spore phenotype was pleiotropic in that sporangiospores capable of germinating on cellobiose could always germinate on glutamate or xylose; but C-spores capable of germinating on xylose or glutamate did not always germinate on cellobiose. There was a hierarchy of substrates capable of initiating germination with glucose = mannose greater than xylose greater than glutamate greater than cellobiose. C spores also differed from M-spores by initiating germination in the presence of the non-metabolizable glucose analogue 3-O-methylglucose. These results suggest that at least two sporangiospore phenotypes are produced depending upon the concentration and type of ingredients present in the sporulation medium. PMID- 6279768 TI - TnA-directed deletion of the trp operon from RSF2124-trp in Escherichia coli. AB - Plasmid pMT-trp was constructed by digestion of RSF2124-trp with restriction endonuclease PstI and ligation with T4 ligase. In pMT-trp about 78% of the DNA of transposon TnA from RSF2124-trp was deleted, and hence the gene for ampicillin resistance was lost. All Trp- segregants from pMT-trp carriers in Escherichia coli W3110 and its derivatives were found to have lost the entire plasmid. On the other hand, deletion plasmids which had lost the trp operon were found among Trp- segregants from RSF2124-trp carriers, particularly from the mutant strain trpAE1 trpR tnaA. The experimental fact that deletion occurred exclusively in RSF2124 trp suggests that the presence of TnA in the plasmid (RSF2124-trp) was responsible for the deletion. PMID- 6279769 TI - Review article initial stages in infection with animal viruses. PMID- 6279770 TI - Use of recombinant plasmids to investigate the structure of the human cytomegalovirus genome. AB - Human cytomegalovirus (HCMV) DNA was digested with restriction endonucleases and the fragments characterized with respect to molecular weight and relative mole proportions. The terminal fragments were identified by digesting HCMV DNA with exonucleases before restriction endonuclease treatment and subsequent gel analysis. The HindIII fragments of HCMV DNA were cloned in Escherichia coli and recombinant plasmids were characterized by digestion with restriction endonucleases and by molecular hybridization with HindIII, Bg/II and XbaI fragments of the virus genome. Data from these experiments were used to construct physical maps of HCMV DNA for the HindIII, Bg/II and XbaI restriction endonucleases. The terminal regions of the genome and the region containing fragment HindIII M were shown to be heterogeneous. PMID- 6279771 TI - Simian foamy virus pseudotypes of vesicular stomatitis virus: production and use in sero-epidemiological investigations. AB - Simian foamy virus (SFV) pseudotypes of vesicular stomatitis virus have been successfully produced and their host range characterized. The availability of these pseudotypes has permitted the development of a rapid, quantitative assay to measure neutralizing antibody titres to SFV that has proved useful in a sero epidemiological study. PMID- 6279772 TI - Measurement of surface antigen by specific bacterial adherence and scanning electron microscopy (SABA/SEM) in cells infected by vesiculovirus ts mutants. AB - Temperature-sensitive (ts) mutants of the rhabdoviruses vesicular stomatitis virus and Chandipura virus have been used to measure the appearance of virus antigen on the surface of infected cells by the technique of surface analysis by bacterial adherence and scanning electron microscopy (SABA/SEM). The number of staphylococci specifically adhering to antiserum-treated infected PTK-2 or BSC-1 cells at permissive (31 degrees C) and restrictive (39 degrees C) temperatures was followed in time-course experiments and a close correspondence was observed between the proportion of staphylococci bound at 39 degrees C and the known phenotypic properties of the ts mutants. Virus surface antigen was undetected in cells infected by transcription- and replication-defective ts mutants with thermolabile L proteins under restrictive conditions up to an input multiplicity of infection of 50, and in cells infected by a replication-defective NS protein mutant. Some surface antigen was detected late in infection in PTK-2 cells infected by a replication-defective N protein mutant. Surface antigen accumulated normally in maturation-defective mutants with lesions in envelope proteins. These results establish the suitability of the SABA/SEM technique for quantitative estimation of virus antigen on the surface of infected cells. PMID- 6279773 TI - Transplacental Japanese encephalitis virus (JEV) infection in mice during consecutive pregnancies. AB - Transplacental transmission of Japanese encephalitis virus (JEV) has been demonstrated in consecutive pregnancies of mice. Pregnant mice inoculated intraperitoneally with JEV transmit the virus to the foetus. When such female mice were mated again after 6 months, the virus could be isolated from the foetuses of the ensuing pregnancy. The incidence of abortion was increased significantly though the neonatal deaths were considerably less than during the first pregnancy. Intra-uterine infection occurred in spite of the presence of HAI antibodies against JEV in the preconceptional sera of the mice. The findings of the present study indicate the value of such a system for further investigations of the pathogenesis of JEV infection during pregnancy in humans. PMID- 6279774 TI - Envelope protein of the flavivirus Kunjin is apparently not glycosylated. AB - The envelope protein E (formerly designated V3) of the flavivirus Kunjin was not labelled with radioactive galactose, mannose or glucosamine during virus growth in Vero cells. On electrophoresis through polyacrylamide gels containing SDS, the envelope (E) protein migrated more rapidly than related intracellular virus specified glycoproteins. Furthermore, E had a density in CsCl solution consistent with that of a protein lacking carbohydrate, and did not bind to concanavalin A agarose. In contrast, the envelope glycoprotein of Murray Valley encephalitis virus (MVEV) did bind to concanavalin A under similar conditions and was readily labelled with radioactive mannose. These results suggested that the E protein of Kunjin virus was not glycosylated, a feature not shared with MVEV and West Nile virus (WNV), whose properties were consistent with the presence of oligosaccharides attached to the envelope proteins. When Kunjin virions were labelled with radioactive glucosamine, the label was contained in GP19 (formerly NV2). The glycopeptides derived by Pronase digestion of GP19 from Kunjin virions were larger than those derived from GP19 obtained from infected cells. PMID- 6279775 TI - Studies on Proteins of simian sarcoma-associated virus with different growth history. AB - Simian sarcoma-associated virus (SSaV) was repeatedly passaged on three human cell lines. The proteins of the progeny virus were analysed for the presence of variant polypeptides. Occasionally, a few variant polypeptides were observed. One dimensional peptide maps of the major virus protein p30 revealed no modifications after 25 cycles of infection on the three cell lines studied. The peptide map of Pr65gag of virus grown through 25 passages on a human chondrosarcoma cell line was slightly different from that of the virus stock before passaging. The relative amount of the virus protein p30 as compared to p18 and p16 (possibly the SSaV equivalents of p15E and p12E) was variable depending on the host cell. Virus grown on Daudi cells was relatively deficient in p18 and p16. These virus particles were morphologically altered and had a low infectivity. PMID- 6279776 TI - Biological characteristics of type C viruses isolated from different Friend erythroleukaemic cells. AB - We have examined the expression of type C RNA viruses in different Friend erythroleukaemic cell types, distinguished on the basis of increasingly malignant characteristics, which arise during the ageing of mice inoculated with the polycythaemic Friend virus complex. Most early appearing Friend cells (type I) expressed ecotropic virus, but cells of later malignant types showed decreased and variable expression. In general, the more malignant cells released less ecotropic virus. Xenotropic virus was detected in low numbers from type I, II, and IV cells. Two viruses were cloned from type II tumour cells: a xenotropic virus (II clone 1) and an N-tropic ecotropic virus (II clone 2). No pathogenic activity was found when II clone 1 was inoculated into newborn and adult DBA/2J and NIH/Swiss mice observed for up to 20 months, whereas II clone 2 caused a rapid anaemic erythroleukaemia in both N- and B-type newborn mouse strains. It caused a similar form of leukaemia in susceptible N-type adult mice, but at a lower frequency and with a longer latency (usually greater than 5 months). This finding demonstrated a lack of NB restriction in newborn mice. The virus was much less active in DBA/2J mice from which it had been originally cloned; it also appeared to cause lymphoma or to shorten the latency of spontaneous lymphoma in DBA/2J mice. PMID- 6279777 TI - Haemagglutination by bovine leukaemia virus. AB - Bovine leukaemia virus (BLV) was found to agglutinate mouse erythrocytes. Under optimal conditions, including the use of neuraminidase-treated erythrocytes, 200 microgram/ml of BLV purified from the supernatant fluid of BLV-infected bat cells had haemagglutinating titres of about 512 units. BLV haemagglutination was drastically affected by pH and temperature; maximum agglutination occurred at pH 6 and 4 degrees C. That the BLV haemagglutinin is a glycoprotein was suggested by the fact that trypsin, potassium periodate or neuraminidase, but not lipid solvents or phospholipase C, significantly reduced the haemagglutinating (HA) activity of purified BLV. Furthermore, purified BLV glycoprotein of mol. wt. 51 000 (gp51) had HA activity. The receptors for BLV on mouse erythrocytes were inactivated by proteolytic enzymes but not by sodium deoxycholate or potassium periodate. Neuraminidase treatment of erythrocytes increase their agglutinability fourfold. Haemagglutination is a relatively sensitive test for detecting BLV glycoprotein because 0.4 microgram/ml of glycoprotein can be detected by this method. The pH and temperature sensitivity of the BLV HA reaction and specificity for mouse erythrocytes distinguish BLV from that of equine infectious anaemia virus and murine leukaemia virus, the other C type retroviruses known to have HA activity. PMID- 6279778 TI - Atypical patterns of neural infection produced in mice by drug-resistant strains of herpes simplex virus. AB - Mice inoculated intracerebrally (i.c.) with a mutant strain of HSV were found to develop cataracts 1 to 2 months after inoculation. Cataract formation was subsequently shown to follow an acute retinitis which commenced within 1 week of inoculation. The mutant had been selected for high resistance to the nucleoside analogue acyclovir and has been shown previously to be defective in the induction of thymidine kinase and also to express an altered DNA polymerase. The LD50 for mice inoculated i.c. was greater than 10(5) p.f.u. compared with approx 7 p.f.u. for the parental strain. Studies of virus replication following i.c. inoculation with a sublethal dose of the mutant revealed that only small amounts of infectious virus were produced in the brain, but during a period from 6 to 12 days after inoculation vigorous replication occurred in retinal tissue, producing very high titres of virus. PMID- 6279779 TI - Detection of human cytomegalovirus- specific IgA antibodies in colostrum by enzyme-linked immunosorbent assay (ELISA). AB - Fifty women were examined after delivery for the prevalence of antibodies to human cytomegalovirus in colostrum and sera. Eighty percent of them had specific CMV IgG antibodies in the sera, as determined by the immunoperoxidase antibody to membrane antigen (IPAMA) technique. Of the CMV-seropositive women, 60% had specific CMV IgA antibodies in high titer in the colostrum as determined by enzyme-linked immunosorbent assay (ELISA). In only two of the seropositive women were specific CMV IgA antibodies detected in the sera as well. The significance of specific CMV IgA antibodies in colostrum as protection against perinatal infection and the mechanism of this production will be discussed. PMID- 6279780 TI - Excretion of hepatitis A virus in the stools of hospitalized hepatitis patients. AB - A study was carried out to determine whether hepatitis A virus (HAV) can be detected in the stools of patients hospitalized for HAV infection. Acute phase samples of whole blood and stool, as well as completed questionnaires, were obtained from 31 patients hospitalized at any of 13 hospitals in the Phoenix metropolitan area. Blood specimens were tested for hepatitis B surface antigen (HBsAg), IgG antibody to HAV (IgG anti-HAV), and IgM antibody to HAV (IgM anti HAV). Stools were tested for HAV by radioimmunoassay. Five patients (16.1%) had acute hepatitis B, five (16.1%) had acute non-A/non-B hepatitis, and 21 (67.7%) had acute hepatitis A. Of these 21 patients with acute hepatitis A, 11 (52.4%) were found to have HAV in their stools. These results confirm the potential for infectivity of stools of patients hospitalized for hepatitis A and emphasizes the need for caution when dealing with such stools. PMID- 6279781 TI - Early replicative block prevents the efficient growth of fastidious diarrhea- associated adenoviruses in cell culture. AB - Fastidious enteral adenoviruses (EAds) recovered from infants with diarrhea were studied to determine the basis for their inability to propagate efficiently in conventional cell lines. By immunofluorescence microscopy, only rare EAd-infected KB and HeLa cells were shown to synthesize detectable levels of virion proteins. Sedimentation of Hirt-extracted DNAs in sucrose gradients and DNA hybridization analyses demonstrated that EAd DNA synthesis is highly restricted in HeLa cells. Some early gene functions seem to be expressed, however, because Eads can help adenovirus-associated viruses (AAV). Thus, EAd replication in conventional cell lines is blocked at an early step in its growth cycle. PMID- 6279782 TI - Relationships between perfusion defects and static brain scan positivity in patients with ischaemic completed stroke: considerations about the origin of the increased uptake. AB - The relation between perfusion defects shown by radionuclide angiography and static brain scan positivity was evaluated in patients with ischaemic completed stroke at various intervals from the onset of symptoms. An inverse relation between radionuclide angiography and static scan positivity was found for the period within 15 days of the onset of symptoms. The possible relation between changes in perfusion and static brain scan positivity is discussed. PMID- 6279783 TI - Agraphesthesia. A disorder of directional cutaneous kinesthesia or a disorientation in cutaneous space. AB - Tests for graphesthesia and for directional cutaneous kinesthesia (DCK) were performed on a large series of neurological patients and normal subjects, in addition to the standard tests for discriminative sensation. Defects in graphesthesia and DCK were found with lesions at all levels of the nervous system. Graphesthesia was more often and more severely affected than DCK. These functions when impaired were always associated with other sensory defects (directional joint kinesthesia, two-point discrimination, etc.) in different combinations. It appears DCK is probably the basis for graphesthesia. Recent experimental studies have provided an anatomic and physiologic basis for DCK and for graphesthesia. These studies have also discredited wide-held beliefs on the transmission of discriminative sensation through the spinal cord. In this light, the history of ideas about sensation and its mediation is reviewed, and it is concluded that DCK alone deserves to be called a "posterior column function". Graphesthesia and DCK are discussed as kinesthetic functions implying orientation in cutaneous sensory space. These are compared to stereognosis and braille reading, which are complex derived functions depending also on motion, but directed towards recognition in external haptic space. Graphesthesia and DCK should both be considered as distinct forms of somatic sensibility which are valuable adjuncts to the clinical sensory examination. PMID- 6279784 TI - Effect of anisomycin on stimulation-induced changes in dendritic spines of the dentate granule cells. AB - Tetanic stimulation of the entorhinal area induces significant enlargement of the average dendritic spine area and perimeter in the middle and distal thirds of the dentate molecular layer 4 and 90 min following stimulation. Four minutes after stimulation, the differences between the stimulated and control animals were 20% for the dendritic spine area and 9% for the perimeter in the middle third, and in the distal third 32 and 14%, respectively. Ninety minutes after stimulation the differences were 28 and 11% for the area and perimeter in the middle third, and 33 and 18% in the distal third, respectively. Anisomycin at a dose of 25 mg/kg had no significant effect on the average spine area or perimeter in the various thirds of the dentate molecular layer in the 19 and 105 min post-application intervals. This dose of anisomycin given 15 min prior to the stimulation suppresses the stimulation-induced spine changes in the 4 min interval. In the 90 min interval when the effect of anisomycin on protein synthesis is largely terminated, spine enlargement reappears, being 21% higher than the controls in the middle and distal thirds. The differential effect of anisomycin on dendritic spines in the two post-stimulation intervals is discussed in relation to the effect of anisomycin on protein synthesis. The present experiments thus demonstrate that the stimulation-induced spine enlargement in the dentate fascia can be suppressed by a protein synthesis blocking drug. PMID- 6279785 TI - Differentiation of identifiable lobster neuromuscular synapses during development. AB - The ultrastructure of physiologically identified low and high release synapses arising from a single axon on fibres of the distal accessory flexor muscle (DAFM) in a mature lobster was examined by serial section electron microscopy. Low release neuromuscular terminals located only on the proximal fibre were characterized by large synapses (mean area 2.084 micron2), small presynaptic dense bars (mean are 0.021 micron2) and hence a low (2.3%) ratio of dense bar area to synaptic area. In contrast high output terminals located only on the distal fibre had smaller synapses (mean area 0.625 micron2), large dense bars (mean area 0.066 micron2) and a high (23.9%) ratio of bar area to synaptic area. A similar ratio was consistently found for each synaptic type in several other examples of mature lobsters. Hence it was used as a criterion for determining the point at which differentiation occurs during development. In the first larval stage (24 h old) the innervation was localized and undifferentiated. In the fourth (2 week old) and twelfth (1 y old) stage lobsters, the innervation had proliferated to small bundles of proximal and distal fibres. During development synapses increase in their mean surface area in the proximal fibre while remaining constant in the distal fibre. The mean surface area of the dense bars is similar in all stages except for the proximal fibres of the twelfth stage where it is smaller by 50%. Similarly the ratio fo dense bar area to synaptic area is not significantly different for all stages except for the twelfth stage proximal fibres where it is half the value. Consequently differentiation of low and high release neuromuscular terminals occurs by the twelfth stage with an increase in the mean surface area of synapses and a decrease in the mean surface area of dense bars. This morphological differentiation is enhanced in the mature lobster. PMID- 6279786 TI - The distribution of orthogonal arrays in the freeze-fractured rat median eminence. AB - The distribution of orthogonal arrays of particles and their relationships to gap and tight junctions have been studied in the glia of the freeze-fractured rat median eminence (ME). These rectilinear clusters of intramembrane particles are thought to represent trans-membrane channels for ions or metabolites, and were found to be densely packed on the membranous laminations of the pial-glial limitans. Additionally, arrays were found to be present on all of the perivascular glial end-feet examined. Two classes of end-feet were distinguished by their relative densities of orthogonal arrays. End-feet displaying low densities of arrays occurred more frequently in the internal zone, while end-feet displaying high densities occurred more often in the external zone. Similar distinctions based on array density could be made in membranes from other regions of the cell as well. Cross-fractures revealing the cytoplasm underlying these membranes often exposed lipid inclusion bodies, suggesting that membranes containing few arrays belong to tanycytes (or to 'astrocyte-like tanycytes'). The distribution of arrays appeared to be unrelated to the distribution of gap junctions in the membranes of astrocytes and tanycytes (and 'astrocyte-like tanycytes') of the ME, appearing near to and far from gap junctions with approximately equal frequency. Orthogonal arrays were absent from glial membranes near synaptic profiles in the ME. Arrays were also absent from the microvillous membranes of the apical surfaces of ependymal cells, from the cytoplasmic protrusions into the CSF of tanycytes, and from the vicinity of the tight and complex junctions linking the tanycyte and ependymal cell lateral membranes near their apical poles. These results suggest that there is a gradient of array density for most glia of the ME, increasing from the ventricular to the pial surface. PMID- 6279787 TI - Tetrodotoxin-resistant propagating action potentials in presynaptic axon of the lobster. PMID- 6279788 TI - Parallel fluctuations of EPSP amplitude and rise time with latency at single Ia fiber-motoneuron connections in the cat. PMID- 6279789 TI - Distributions of EPSP latency at different group Ia-fiber-alpha-motoneuron connections. PMID- 6279790 TI - Drosophila mutants with opposing effects on nerve excitability: genetic and spatial interactions in repetitive firing. PMID- 6279791 TI - Factors affecting cutaneous mechanoreceptor response. I. Constant-force versus constant-displacement stimulation. PMID- 6279793 TI - Retinal Y-cell activation of deep-layer cells in superior colliculus of the cat. PMID- 6279792 TI - Factors affecting cutaneous mechanoreceptor response. II. Changes in mechanical properties of skin with repeated stimulation. PMID- 6279794 TI - Laminar organization of receptive-field properties in lateral geniculate nucleus of bush baby (Galago crassicaudatus). PMID- 6279795 TI - Norepinephrine regulation of cerebral glycogen utilization during seizures and ischemia. AB - Norepinephrine (NE) depletion of the cerebral cortex after lesion of the ipsilateral locus ceruleus (LC) causes abnormalities of cerebral oxidative metabolism when the cortex is stimulated to increased energy demand (Harik, S. I., J. C. LaManna, A. I. Light, and M. Rosenthal (1979) Science 206: 69-71; LaManna, J. C., S. I. Harik, A. I. Light, and M. Rosenthal (1981) Brain Res. 204: 87-101). These abnormalities were exhibited as decreased mitochondrial reducing equivalent flow. One possible cause of this would be the decreased availability of oxidative metabolic substrates in the NE-depleted cortex. We therefore investigated the effect of unilateral LC lesion and the resultant depletion of ipsilateral endogenous NE on glycogen and other energy metabolites in the cerebral cortex of rats under three conditions: (1) at "rest," (2) when energy demand is inncreased markedly by seizures, and (3) during total cerebral ischemia. We report no differences in cerebral metabolites between NE-depleted and control hemispheres at "rest." In seizures and ischemia, however, the increase in the level of adenosine 3':5'-monophosphate (cyclic AMP) and the breakdown of glycogen were impaired considerably in the NE-depleted cortex. The data suggest that depletion of central NE impairs cerebral glycogenolysis in response to increased energy demands and ischemia. Such impairment may be mediated via a cyclic AMP-related mechanism. PMID- 6279796 TI - Dopamine action on hippocampal pyramidal cells. AB - Dopamine (DA) was applied to CA1 region pyramidal cells in slices of guinea pig hippocampus maintained in vitro in order to examine its electrophysiological effect on CNS neurons. DA induced hyperpolarization of membrane potential and an increased conductance in 75% the 21 CA1 neurons to which it was applied. DA also augmented the afterhyperpolarizations and increased conductance which normally follow spike trains in these neurons. These effects were not altered by intracellular injections of Cl- but were blocked when slices were bathed in Mn2+ solutions. The Mn2+ blockade of DA-induced hyperpolarizations could be overcome when large amounts of agonists were applied. The DA effects were long lasting, were mimicked by the dopamine agonists apomorphine and Epinine, and were blocked by the dopamine antagonists flupenthixol and chlorpromazine. Extracellular or intracellular application of cyclic AMP mimicked the effects of DA. The results suggest that DA-induced hyperpolarization and conductance changes are mediated by a Ca2+-activated K+ conductance. DA may increase the intracellular Ca2+ concentration through effects on one of the Ca2+ buffering mechanisms. The long duration of these effects suggest that DA works though some intracellular intermediary, perhaps cyclic AMP, considering that the actions of cyclic AMP on membrane properties are similar to those of DA. The dopaminergic projection to the hippocampus should have a powerful inhibitory action, which would be most effective in modulating the activities of neurons exhibiting high levels of excitability, particularly cells involved in cyclical burst generation. PMID- 6279797 TI - Cannabinoids inhibit calcium uptake by brain synaptosomes. PMID- 6279798 TI - Pseudopodial interdigitations between abutted nerve terminals: diffusion traps which occur in several nuclei of the rat limbic system. AB - Stimulation of the Torpedine ray electric organ can cause the loss of synaptic vesicles and the growth of pseudopodia from the nerve terminals (Boyne, A. F., and S. McLeod (1979) Neuroscience 4: 615-624). The latter embed themselves in corresponding indentations in abutted terminals. The geometry of these pseudopodial indentations (PSIs) can vary: (i) in length, (ii) in the extent of constriction of the base, and (iii) through a compound interaction between different pseudopodia extending in opposite directions. Examination of six rat brain nuclei in the limbic system has shown that their neuropil can be categorized according to the prevalence of either (i) nerve terminals indented by nerve terminal outgrowths (i.e. PSIs) or (ii) nerve terminals indented by dendritic outgrowths: these have been previously termed spinules. Clusters of simple PSIs were seen in the central nucleus of the amygdala, while base constricted and compound forms were found in the globus pallidus and substantia nigra. Dendritic spinules were prevalent in the nucleus accumbens and the molecular layer of the hippocampus. In the CA4 hilar region of the hippocampus, large nerve terminals containing PSIs were found. The caudate neuropil appeared to be of mixed character in that the small terminals often had spinules but occasionally showed PSIs. Spinules have been recognized for many years and the possibility of their plasticity has been raised previously (Tarrant, S. B, and A. Routtenberg (1977) Tissue Cell 9: 461-473). The present report appears to be first detailed description of an alternative form of invasion which is known to be plastic in the elasmobranch electric organ. It is suggested that the extracellular space between the partners of a PSI could act as variable diffusion traps. If the involved boutons carry action potentials, then nonsynaptic release and accumulation of substances such as potassium, amino acids, and nucleotides may be expected during stimulation. Consequent direct or receptor-mediated effects on the membrane potential could influence transmission through adjacent synapses. PMID- 6279799 TI - Orientation of neurite growth by extracellular electric fields. AB - Extracellularly applied steady electric fields of 0.1 to 10 V/cm were found to have marked effects on the neurite growth of single dissociated Xenopus neurons in culture: (1) neurites facing the cathode showed accelerated growth, while the growth of those facing the anode was reduced. Neurites growing relatively perpendicular to the field axis were prompted to curve toward the cathode. (2) More neurites appeared to be initiate from the cathodal side of the cell. (3) The number of neurite-bearing neurons per culture and the average neurite length were increased. These effects are absent in cultures treated with electric fields of similar strength but alternating polarity and cannot be attributed either to a gradient of extracellular diffusible substances or to the flow of culture medium produced by the field. The field effects are reversible: (1) removal of the electric field resulted in the loss of neurite orientation in a few hours and (2) reversal of the polarity of the electric field led to a rapid reversal in the neurite orientation. To determine the cellular loci of these field effects, we treated the neurons with a number of pharmacological agents or altered their ionic environments. Incubation with concanavalin A (Con A) was found to abolish these filed effects completely. Since the binding of Con A to the neuronal surface was shown to prevent field-induced accumulation of the Con A receptors toward the cathodal side of these neurons, our finding is accumulation of the Con A receptors toward the cathodal side of these neurons, our finding is consistent with the notion that cathodal accumulation of growth-controlling surface glycoproteins by the field is the underlying mechanism of the field-induced orientation of neurite growth toward the cathode. PMID- 6279800 TI - Impulse activity in presynaptic dendrites: analysis of mitral cells in the isolated turtle olfactory bulb. AB - Impulse activity has been reported in neuronal dendrites in several regions of the central nervous system, where it is believed to assist in boosting transmission of signals from remote dendritic sites to the cell body. We have studied this activity in the dendrites of mitral cells in an isolated preparation of the turtle olfactory bulb. Intracellular recordings have been obtained from mitral cells responding to single volleys in the olfactory nerves or lateral olfactory tract. In addition to the large somatic spike, a small fast prepotential (FPP) was present in nearly all cells in response to an orthodromic volley in the olfactory nerves, but it was never seen in antidromic responses from the lateral olfactory tract. Collision tests using antidromic and orthodromic volleys showed that the EPP does not propagate into the axon. Hyperpolarizing current injections caused delay and blocking of the soma spike with little effect on the FPP response. These and other tests provided evidence to localize the EPP in the dendrites and to distinguish it from injury potentials and from spikes in the axon hillock or axonal initial segment. These results suggest that one function of the impulse in mitral cell dendrites is the classical one of boosting transmission of synaptic responses from the glomerular tuft to the cell body. In addition, it si well established that mitral cell dendrites are presynaptic to the dendrites of interneurons within the bulb and that these connections provide pathways for recurrent inhibition of the mitral cells. It therefore appears that the dendritic impulse in mitral cells acts as a booster for local dendritic synaptic output. These results provide further evidence for the multiple state-dependent input-output functions of cells with presynaptic dendrites. PMID- 6279801 TI - A modified method for the in vivo labeling of red blood cells with Tc-99m: concise communication. AB - The rate of incorporation of Tc-99m into red blood cells pretinned in vivo was measured by collecting blood samples in stannous DTPA solution, which served as a competing ligand for Tc-99m. This collection technique permitted a measurement of high-affinity red-cell labeling efficiency at the instant of sampling. At 0.5 min after injection only 62% of technetium is tightly bound to the red cell; this rises to 94.5% at 10 min. Based on the graded labeling of the red cells, the in vivo labeling procedure was modified by isolating pertechnetate and red blood cells tinned in vivo in a syringe during the first 10 min of labeling. The pertechnetate is thus prevented from distributing to extravascular compartments, and 90% of the injected Tc-99m is firmly bound to red blood cells at the time of injection. In a series of 23 patients, seven were tested with the in vivo method and seven with the modified in vivo method, and nine patients were tested with each method on separate occasions. A decrease in gastric activity and improved image quality were found with the modified method compared with the standard method of in vivo red-cell labeling. PMID- 6279802 TI - Radiochemical analysis of Tc-99m human serum albumin with high-pressure liquid chromatography: concise communication. AB - High-pressure liquid chromatography (HPLC) can be performed with an aqueous size exclusion column to separate proteins or other macromolecules on the basis of molecular size. An HPLC system with a Spherogel-TSK SW column was modified to detect simultaneously uv absorption and radioactivity. Characteristic retention times (RT) were determined for pure human serum albumin (HSA) (RT = 17 min) and pertechnetate (RT = 28.5 min). When analysis was performed on Tc-99m HSA preparations, Tc-99m radioactivity was resolved into five different peaks, with RT ranging from 10.2 to 28.5 min. Less than 2% radioactivity was associated with the pertechnetate peak, whereas the remaining Tc-99m was protein bound. Most of the activity (90%) corresponded to the albumin peak, and 7% was bound to contaminants of high molecular weight with RTs of 10.2 and 14 min. Rapid separation of various radiochemical components differing in molecular size provides an improved basis for understanding the biodistribution of a Tc-99m HSA preparation. This technique would be useful for the preparation and analysis of various radiolabeled macromolecules such as enzymes, immunoglobulins, and other proteins. PMID- 6279803 TI - Effects of dietary long-chain fatty acids on the rat biosynthesis of unsaturated fatty acids in the rat. AB - Menhaden oil (ME) was included in semisynthetic diets to study the effect of long chain fatty acids, mainly 20:5n3 and 22:6n3, on the biosynthesis of polyunsaturated fatty acids and on the 6- and 9-desaturase activities in liver microsomes. Five experimental diets, with the following fat supplements, were fed to male rats from weaning for a period of 33 weeks: 5% safflower oil (SAF) + 10% hydrogenated coconut oil (HCO), 5% SAF + 5% HCO + 5% ME, 5% SAF + 10% ME, 15% HCO and 5% HCO + 10% ME. The last two diets were deficient in linoleic acid. The three nondeficient diets contained similar amounts of linoleic acid. Including ME in the diets depressed the 6- and 9-desaturase activities, especially in the linoleic acid--deficient rats. The synthesis of 20:4n6, 22:4n6 and 22:5n6 were depressed. These effects were related to the preferential accumulation of dietary 20:5n3 and 22:6n3 in the liver microsomes, as compared to the n6 fatty acids. It is hypothesized that dietary 20:5n3 and 22:6n3 increase the minimum requirement for linoleic acid in the diet. PMID- 6279804 TI - Utilization of bamboo by the giant panda. AB - Two giant pandas were used to assess the utilization of bamboo as a feedstuff. Three 1-week-long digestion trials were conducted during which feed intake and fecal output were recorded. Passage of digesta was measured from both fluid and particulate markers administered at feeding. Results indicate that the giant panda, although highly specialized for the consumption of bamboo, is a very inefficient digester of bamboo. The pandas consumed up to 6% of body weight in dry matter per day, with bamboo dry matter digestibility averaging less than 20%. Apparent digestion coefficients for the structural carbohydrates of bamboo (27% for hemicellulose and 8% for cellulose) indicate that, unlike most herbivores, pandas do not rely heavily on the microbial degradation of plant material to fulfill their nutritional requirements. Additionally, the passage of digesta through the gastrointestinal tract of the panda was extremely rapid, with complete clearance of markers in less than 12 hours. The giant panda seems to have specialized to a feed source high in plant fiber without extensive modification of the digestive tract by selectivity in feeding, effective mastication, ingestion of large quantities, digestion of cellular contents rather than cell walls and rapid fecal excretion of the undigested residue. PMID- 6279805 TI - Effect of dietary fiber components on fecal nitrogen excretion and protein utilization in growing rats. AB - The effects of purified fiber components and wheat bran on several indices of protein utilization were determined in growing rats. A control diet containing 10% casein was diluted with either cellulose (C), pectin (P), lignin (L), guar gum (G), or wheat bran (W) at fiber levels ranging from 3% to 20%. All fibers except C caused a decrease in net protein ratio (NPR) as compared to the control casein diet. This depression in NPR increased as the dietary fiber level increased. Apparent and true nitrogen digestibilities also decreased with all fibers at all levels. At the highest level of fiber (20%) the depression was greater for G and W and was least for C. NPR when divided by digestibility (analogous to biological value) decreased with P,L, W (all levels) and G (20% level) but not with C. When rats were fed fiber without protein, there was increased excretion of endogenous fecal nitrogen with all fibers at all levels. The results demonstrate that fiber(s) affected protein utilization as measured by NPR, digestibility and endogenous fecal nitrogen excretion and that the negative effect increased with the level of fiber consumed. PMID- 6279806 TI - Vitamin D compounds in cows' milk. AB - The milk from cows fed normal levels of vitamin D has been found to contain approximately 40 IU per liter of vitamin D activity. A 14-fold increase in dietary vitamin D intake causes only a doubling of the amount of vitamin D in milk. This was determined by measuring stimulation of intestinal calcium transport in the vitamin D-deficient rat. Four vitamin D compounds were then isolated from cow's milk using a combination of conventional chromatography on Sephadex LH-20 and Lipidex 5000 followed by high-performance liquid chromatography. 24,25-Dihydroxycholecalciferol and 1,25-dihydroxycholecalciferol were measured using binding protein assays. One liter of milk contained 27 ng and 4.9 ng, respectively, of these two metabolites. Together these account for about 15% of the vitamin D activity. Cholecalciferol was found to be present at a concentration of 281 ng/liter or 11 IU/liter of biological activity. The milk contained 145 ng/liter 25-hydroxycholecalciferol or 29 IU/liter of activity. Therefore the known vitamin D compounds fully account for the biological activity observed in milk. It is therefore clear that no evidence could be found for the existence of a highly active water-soluble form of vitamin D in milk. PMID- 6279807 TI - Hepatic gluconeogenic enzymes, plasma insulin and glucagon response to magnesium deficiency and fasting. AB - Three experiments were conducted to assess the effects of magnesium deficiency on the activities of hepatic glucose-6-phosphatase (G6Pase), fructose 1,6 bisphosphatase (FDPase) and phosphoenolpyruvate carboxykinase (PEPCK). Experiment 1 was designed to determine if magnesium deficiency interfered with the gluconeogenic response to fasting. Rats were fed either a control (C) or magnesium-deficient (MD) diet for 12 days. One-half of each group of rats was fasted for 24 hours prior to death. Hepatic enzyme activities, plasma and liver magnesium, and whole blood glucose were measured. Activities of G6Pase and PEPCK were higher in fasted group C rats compared to fed group C rats. Activity of FDPase was lower. The response was similar in the MD groups. Comparison of C and MD groups indicated that magnesium deficiency was accompanied by an increase in PEPCK activity. To verify this result and to investigate the role of anorexia in producing increased PEPCK activity, experiment 2 included a pair-fed group (PF). The results indicated that anorexia was not responsible for increased PEPCK activity in MD rats. The relation of circulating insulin and glucagon concentrations to effects of magnesium deficiency was explored in experiment 3. A decreased insulin:glucagon ratio was observed in MD rats. The results of these experiments suggest that magnesium deficiency alters PEPCK activity by affecting secretion of pancreatic hormones. PMID- 6279808 TI - Effects of dietary fiber on the bioavailability of folic acid monoglutamate. AB - Low bioavailability of folacin has been previously reported for a variety of foods of plant origin. This study was conducted to examine the possible role of various types of dietary fiber on the bioavailability of folic acid monoglutamate. Cellulose, pectin, lignin, sodium alginate and wheat bran were selected for their differing physical and chemical properties. In vitro binding studies by equilibrium dialysis showed no evidence of physical or chemical binding of folic acid under physiological conditions. In vivo effects were evaluated by a chick bioassay with graded levels of folic acid in semipurified diets containing the fiber materials at 3% (weight/weight). Total liver and plasma folacin concentration and chick growth were used as response indicators. Dose-response curves indicated that pectin, lignin and alginate significantly reduced chick growth at all levels of dietary folacin. Plasma and liver folacin dose-response curves were not significantly different for any of the fiber materials, which indicated that the growth impairment was not due to a fiber effect on folic acid absorption. These results suggest that added dietary fiber has little or no effect on the bioavailability of folic acid monoglutamate. PMID- 6279809 TI - The radiologic diagnosis of Meckel's diverticula. PMID- 6279810 TI - Orbital roof reconstruction with a hydroxyapatite implant. PMID- 6279811 TI - The mesothelium: a cytochemical study of "activated" mesothelial cells. AB - The cytochemical profile of activated mesothelial cells differs from resting cells. The reaction products of enzymes associated with oxidative mechanisms of cell respiration were easily displayed in activated mesothelial cells; in resting mesothelial cells, only enzymes of the pentose pathway were readily demonstrable. Acid hydrolases were detected in greater quantity than in resting cells, possibly reflecting an increased potential for endocytosis. In addition, the cytochemical assays indicated increased Golgi activity was reflected by the demonstration of thiamine pyrophosphatase, while the content of ribonucleic acid was also increased. These cytochemical features compare well with the ultrastructure of active mesothelial cells which possess abundant ergastoplasm and a well-developed Golgi apparatus. In comparison with peritoneal macrophages, activated mesothelial cells differ mainly in the quantity of reaction product, there being more in macrophages. The results were significantly different only in the demonstration of lipids which were never found in mesothelial cells, but were invariably present in macrophages. It is still unclear whether the acid mucopolysaccharide hyaluronic acid is produced by or merely transported through the mesothelium from the subserosal site. PMID- 6279812 TI - Concurrent outbreaks of rhinovirus and respiratory syncytial virus in an intensive care nursery: epidemiology and associated risk factors. AB - An outbreak of viral respiratory disease occurred in eight infants in a neonatal intensive care unit during the 1980 winter respiratory season. Four infections with respiratory syncytial virus and four infections with rhinovirus were identified. Epidemiologic investigation revealed that viral respiratory infection was significantly associated with intubation with orotracheal tubes (P = 0.001), with the presence of both a nasal feeding tube plus an orotracheal tube together (P = 0.007), and with assisted ventilation (P = 0.009) when compared to uninfected controls. Twenty-seven of 85 (30.6%) personnel working in the unit at the time of the outbreak reported a history of upper respiratory illness during the week prior to the outbreak, and 46 (54.1%) of them had had contact with patients in areas of the hospital where patients infected with RSV and rhinovirus were housed. The data suggest that both viruses were transmitted to the babies by hospital personnel. Rhinoviruses can be nosocomial pathogen in neonates with compromised pulmonary function, and the clinical presentation of rhinovirus infection in neonates may be difficult to distinguish from that produced by RSV. PMID- 6279814 TI - Knot integrity of coated synthetic absorbable sutures used for extraocular muscle surgery in a rabbit model. AB - Knot integrity of a new coated polyglycolic acid suture (Dexon Plus) was compared to that using uncoated polyglycolic acid (Dexon-S) and coated polyglactin-910 (Vicryl) for extraocular muscle recessions in rabbits. Knot integrity was found to be virtually identical using 6-0 sutures of any of these materials. Top throws of 5-0 coated Dexon or 5-0 coated Vicryl had a tendency to loosen in a few cases, though this was eliminated in later operations by more careful tightening of knots and avoidance of cutting suture ends to a length less than 2 mm. Subjectively observed differences in "tissue drag" were insignificant in this model. PMID- 6279813 TI - A non-x-linked syndrome with susceptibility to severe Epstein-Barr virus infections. AB - Three siblings developed severe (two) or fatal (one) infectious mononucleosis. This family differed from previously described kindreds with a susceptibility to overwhelming Epstein-Barr virus infections in that: (1) both males and females were affected; (2) they had a history of the recurrent bacterial infections; (3) they produced the full spectrum of antibodies to EBV in the expected range of titers; and (4) survivors recovered completely. Two of these youths, but not their parents or an unaffected sibling with mild IM, had a deficiency of natural killer activity that did not respond to preincubation of their peripheral blood mononuclear cells with interferon. NK activity may have an important role in controlling infections with EBV. PMID- 6279815 TI - Congenital hypertrophic pyloric stenosis and associated anomalies in the genitourinary tract. AB - Genitourinary anomalies were looked for in patients with congenital hypertrophic pyloric stenosis. In a prospective series of 64 patients investigated by intravenous pyelography, 13 were abnormal (20.6%). In a retrospective series of 232 patients, 6 had anomalies of the upper urinary tract (2.7%). In this latter series the incidence of inguinal hernia (3.4%), undescended testes (3.0%), and hypospadias (0.9%) was determined. In another 10 patients urinary tract anomalies (5), urinary infection (2), and a significant family history (3) were found associated with congenital pyloric stenosis. As the incidence of these anomalies is greater than expected, which suggests an interrelationship, a hypothesis has been proposed linking genetic factors and the metabolism of gastrin with the etiology of congenital hypertrophic pyloric stenosis. PMID- 6279816 TI - Pediatric surgical patients with severe anaerobic infection: report of 16 T antigen positive cases and possible hazards of blood transfusion. AB - Red blood cells become polyagglutinable when the normally latent T-antigens of the red blood cell membrane are exposed. Unmasking of T-antigens results from removal of N-acetyl-neuraminic acid by neuraminidase, an enzyme commonly produced by a variety of bacteria. Red blood cells altered in this way are said to be T activated. T-activated red blood cells can be agglutinated by anti-T, an antibody normally present in human serum, so that severe transfusion reactions may occur and have occurred, if T-antigen positive patients are transfused with normal whole blood or plasma. This can be avoided by transfusing only packed or washed red blood cells. From October 1978 to October 1980 we found T-activation in 16 pediatric surgical patients aged 3 days to 14 yr with severe anaerobic infections. This included patients with necrotizing enterocolitis, perforated appendicitis, megacolon, infected anal atresia and gas gangrene. The isolate neuraminidase-producing bacteria were Clostridium perfringens and Bacteroides fragilis. Clinical data of these 16 patients are briefly reviewed and the importance of T-antigen positivity for their management is discussed. PMID- 6279817 TI - Synthesis and characterization of iodoazidobenzylpindolol. AB - A high affinity beta-adrenergic ligand, iodoazidobenzylpindolol, was synthesized and characterized. The absorption spectrum of this compound changed markedly upon photolysis, consistent with decomposition of the azide group. This compound has a KD of 5--7 x 10(-10) M for the duck erythrocyte ghost beta-adrenergic receptor when measured in a competitive binding assay. PMID- 6279818 TI - Separation of penicillin and its major degradation products by ion-pair reversed phase high-pressure liquid chromatography. AB - An ion-pair reversed-phase high-pressure liquid chromatographic technique capable of separating penicillin and its major degradation products within 8 min was developed. The influence of pH, counterion concentration, buffer concentration, and organic modifier content was studied and the observed behavior of the compounds during the chromatographic process was discussed. PMID- 6279819 TI - Interaction of theobromine with sodium benzoate. AB - The interaction of theobromine with sodium benzoate was investigated by PMR spectroscopy. The interaction of theobromine with pentadeuterated benzoic acid (benzoic acid-d5) was examined in the same manner but to a lesser degree. Chemical shifts of theobromine protons were determined as a function of sodium benzoate concentration in deuterium oxide at 30 and 15 degrees. Signals of both methyl groups of theobromine underwent significant upfield shifts when sodium benzoate was added to a theobromine solution. This fact suggests that a complex is formed by vertical stacking or plane-to-plane stacking. The same results were obtained for benzoic acid-d5. PMID- 6279820 TI - Modifications to synaptic transmission at group Ia synapses on cat spinal motoneurones by 4-aminopyridine. AB - 1. The average amplitude of e.p.s.p.s evoked in cat spinal motoneurones by impulses in single group Ia afferents usually increased following the intravenous injection of 4-aminopyridine (4-AP). Most of this increase occurred over the first 30 min following injection of 4-AP. 2. The increase in the average amplitude following 4-AP occurred by a reduction in the probability of occurrence of component e.p.s.p.s with smaller peak amplitudes, and an increase in the probability of occurrence of component e.p.s.p.s with larger peak amplitudes. There was no evidence that the discrete amplitudes of components after 4-AP were a result of graded increases of the discrete amplitudes before 4-AP. 3. The interpretation suggested for these results is that each component e.p.s.p. is generated by transmission at a different combination of boutons. At each of these boutons sufficient transmitter is released to saturate all available receptors. The effect of 4-AP is to decrease the probability of failure to release transmitter at each bouton, including some boutons which, before 4-AP, did not release transmitter. PMID- 6279821 TI - Chemical modification of sodium channel surface charges in frog skeletal muscle by trinitrobenzene sulphonic acid. AB - 1. We have investigated the effects of externally applied trinitrobenzene sulphonic acid (TNBS) on sodium currents of voltage-clamped frog skeletal muscle fibres. TNBS is a membrane-impermeant reagent which reacts specifically with amino groups under physiological conditions. 2. TNBS shifts the voltage dependence of steady-state sodium current inactivation (h infinity) by approximately 18 mV to more hyperpolarized potentials when measured at pH 9. This effect of TNBS is irreversible, suggesting that the reagent permanently modifies some membrane component(s). 3. Time constants for the development of and recovery from inactivation of sodium current are similarly shifted to more hyperpolarized potentials by TNBS treatment. External TNBS did not affect the completeness of sodium current inactivation during depolarizing pulses. 4. The activation of sodium current is less affected by TNBS than is inactivation. The voltage dependence of peak sodium current activation was shifted by only approximately 4 mV by TNBS treatment at pH 9. The kinetics of sodium current activation during depolarization were little affected by TNBS. 5. The TNBS-induced shift in h infinity and sodium current activation are independent of pH when measured at pHs between 7.4 and 11. 6. The results are consistent with the hypothesis that TNBS increases the negative surface charge of the membrane by reacting with membrane amino groups having a high pK. PMID- 6279822 TI - Functional maturation of motor nerve terminals in the avian iris: ultrastructure, transmitter metabolism and synaptic reliability. AB - 1. The transformation of easily fatigued embryonic neuromuscular junctions into highly reliable mature terminals was examined by studying functional and morphological changes during development of the avian iris. The mature ability to follow repetitive electrical nerve stimulation was correlated with the rate of acetylcholine (ACh) synthesis and choline uptake, and with the fine structure of the nerve terminals and the post-synaptic elements.2. The terminals of the ciliary nerve of the chick initially form functional synaptic contacts with the iris muscle at embryonic St. 34-40. At the onset of this period, no Na(+) dependent high affinity choline uptake can be demonstrated, and the low level of ACh synthesis present is sensitive to Na(+) removal. At St. 36 [(3)H]ACh synthesis begins to increase, the increment being Na(+)-dependent.3. ACh synthesis in the embryonic iris was insensitive to a conditioning [K(+)](o) depolarization even as late as St. 43. Just before hatching, depolarization elicits some augmentation in synthesis, but by 2 days ex ovo this release-induced response has increased by an order of magnitude.4. Concurrently with the acquisition of the ability to respond to depolarization with accelerated synthesis, neuromuscular transmission in the iris becomes reliable and secure during stimulation at 20 Hz. Embryonic junctions rapidly block during such stimulation, and the failure is shown to be presynaptic in origin, resulting most probably from failure to sustain adequate levels of transmitter release.5. Ultrastructural examination of the developing ciliary terminals revealed few synaptic vesicles at early stages, and a dearth of other specializations. The sequence of development from these small structurally undistinguished endings to large en plaque junctions completely filled with vesicles was reconstructed and compared to other neuromuscular junctions. Morphological maturation appears progressive with little evidence of discontinuity signalling functional status, but it is only after the terminals enlarge and become closely packed with vesicles that mature synaptic reliability is found.6. The temporal correlation between responsiveness of transmitter synthesis to depolarization and reliable neuromuscular transmission suggests that modulation of neurotransmitter metabolism in response to demand signals the achievement of junctional maturity. PMID- 6279823 TI - Single glutamate-activated channels recorded from locust muscle fibres with perfused patch-clamp electrodes. AB - 1. Glutamate-activated single channels have been examined with conventional and internally perfused patch-clamp electrodes applied to the extrajunctional membrane of locust muscle fibres which were usually treated with concanavalin A to reduce desensitization. Channels opened by glutamate and other agonists have been compared.2. Recording patches were selected where there appeared to be only one active channel under the pipette. The conductance for single glutamate activated channels was 150 pS and was not markedly dependent on clamp potential. The lifetimes of the channels were usually exponentially distributed with a mean of tau(glutamate) = 2.3 +/- 0.12 msec, T = 23 degrees C, V(m) = -60 mV.3. Channels opened by fluoroglutamate had a mean lifetime of tau(fluoroglutamate) = 1.4 +/- 0.1 msec; channels opened by quisqualate had a mean lifetime of tau(quisqualate) = 6.4 +/- 1.0 msec. The conductances of channels opened by fluoroglutamate, quisqualate and glutamate were not significantly different.4. The behaviour of individual receptor-channel complexes has been examined at various concentrations of glutamate. Drug solutions were applied through an internal perfusion pipette which allowed exchange of the solution in the patch electrode tip within 10 sec. The distribution of channel closed times could be fitted with a single exponential. Channel lifetime was not markedly dependent on glutamate concentration (30-600 mum) whereas the channel closed time decreased with increasing glutamate concentration.5. The reciprocal of channel closed time vs. glutamate concentration had a slope value of 1.85 on logarithmic co ordinates. The approximately second power dependence of net forward reaction rate on glutamate concentration suggests that at least two glutamate molecules activate a single receptor-channel complex.6. The apparent dissociation constant for the glutamate-receptor complex is large, being about 300-500 muM. If the receptors have an equally low affinity for neurally released transmitter, then only a small amount of the transmitter packet is expected to bind to receptors. Quisqualate and glutamate have similar receptor affinities whereas receptor affinity for fluoroglutamate is smaller. PMID- 6279824 TI - The effect of theophylline on intestinal bicarbonate transport measured by pH stat in Amphiuma. AB - 1. The influence of theophylline on the mucosa to serosa and serosa to mucosa fluxes of HCO3- were measured by the pH stat technique in isolated segments of proximal small intestine from Amphiuma maintained under short-circuited conditions. The mucosal or serosal fluid was exposed to media containing 25 mM HCO3- (pH 7.4) while the pH of unbuffered media in the opposite compartment was maintained by addition of acid. 2. Theophylline significantly increased the secretory flux of HCO3- and significantly reduced the absorptive flux when measured in Cl- -free (SO4(2-)) media. 3. In normal media theophylline did not alter the secretory flux but significantly lowered the absorptive flux of HCO3-. 4. Acetazolamide (0.1 mM) inhibited the theophylline-stimulated secretory flux of HCO3- and reduced the effect of theophylline on the absorptive flux. 5. In normal intestine there was an inequality between the secretory or absorptive HCO3- flux and the short-circuit current (Isc) consistent with the presence of Cl- absorption. After addition of theophylline the Isc was more nearly equal to the net secretory or absorptive HCO3- flux. 6. Exogenous cyclic AMP had effects identical with theophylline. 7. The results provide strong evidence that elevation of cyclic AMP stimulates net HCO3- secretion in urodele small intestine and provide indirect evidence that Cl- absorption is simultaneously reduced. PMID- 6279825 TI - Electrophysiological and freeze-fracture studies of changes following denervation at frog neuromuscular junctions. AB - 1. Changes which occur at frog neuromuscular junctions following denervation have been studied by combining intracellular recording and freeze-fracture electron microscopy. 2. Shortly after nerve section, both neuromuscular transmission and intramembrane structures of neuromuscular junctions remain normal. 3. Later, neuromuscular transmission fails, beginning with the disappearance of end-plate potentials (e.p.p.s) and followed by the disappearance of miniature end-plate potentials (m.e.p.p.s). The frequency of m.e.p.p.s which persist after cessation of e.p.p.s is not increased dramatically in K+-rich or hypertonic solutions. 4. Concomitant with the changes of transmission are changes in intramembrane structures. The first sign of these changes in disruption of active zones, which become disorganized, fragmented or vanish. Nerve terminals then disintegrate and eventually are engulfed by Schwann cells. 5. When neuromuscular transmission has failed completely, former sites of the neuromuscular junction are occupied by Schwann cells. These cells develop transverse ridges which lie opposed to junctional folds, just like active zones of nerve terminals. However, the ridges on Schwann cells do not contain organized rows of particles or clusters of any synaptic organelles, even at later stages when Schwann cell m.e.p.p.s commence. 6. It is suggested that the failure of e.p.p.s involves at least an impairment of the transmitter release mechanism at the nerve terminal, which is probably associated with the disruption of active zones. The cessation of m.e.p.p.s is thought to be caused by the engulfment of terminals by Schwann cells. PMID- 6279826 TI - The components of synaptic potentials evoked in cat spinal motoneurones by impulses in single group Ia afferents. AB - 1. Excitatory post-synaptic potentials (e.p.s.p.s) were evoked in cat spinal motoneurones by impulses in single group Ia afferent fibres. The probability density of the fluctuations in peak amplitude of each e.p.s.p. was calculated from the recorded peak amplitude and the probability density of the recording noise. 2. Most e.p.s.p.s fluctuated between different components (i.e. individual e.p.s.p.s of a particular discrete amplitude) with peak amplitudes which were integer multiples of the increment between successive components. The average peak amplitude of this incremental e.p.s.p. was about 90 microV for e.p.s.p.s generated at or near the soma. 3. In general, the probability density of the peak amplitude could not be described using Poisson or binomial distributions. 4. For many e.p.s.p.s the complete time course of each component could be calculated. There was no variability in the amplitude of these components nor in their latency of onset. For some e.p.s.p.s there were differences in the latency and time course of the components. 5. The increments between successive components of e.p.s.p. generated proximally were no larger (at the soma) than the corresponding increments for e.p.s.p.s generated at more distal dendritic sites. 6. These results and those from subsequent papers (Jack, Redman & Wong, 1981; Hirst, Redman & Wong, 1981) reinforce earlier suggestions that each bouton behaves in an all-or-nothing manner with respect to post-synaptic effect, and the probability of failure varies at different boutons arising from the same afferent. PMID- 6279827 TI - Post-tetanic potentiation and facilitation of synaptic potentials evoked in cat spinal motoneurones. AB - 1. Excitatory post-synaptic potentials (e.p.s.p.s) were evoked in spinal alpha motoneurones of the cat by impulses in single group Ia nerve fibres. 2. The average peak amplitude of some of these e.p.s.p.s was increased by a conditioning tetanus. The maximum increase observed was 54% of the control amplitude. 3. The average peak amplitude of some e.p.s.p.s was increased by a single conditioning stimulus which preceded the test stimulus by 1 or 2 msec. The maximum increase observed was 28% of the control amplitude. 4. The ability of e.p.s.p.s to potentiate following a tetanus was correlated with their ability to be facilitated by a single conditioning stimulus. 5. If an e.p.s.p. could be facilitated prior to a tetanus, the amount of facilitation was reduced after the tetanus, with all facilitation being abolished when post-tetanic potentiation was maximal. 6. The fluctuations of an e.p.s.p. were analysed before and after a tetanus. The peak amplitudes that an e.p.s.p. fluctuated between while potentiated did not gradually diminish as the effect of the tetanus disappeared. Post-tetanic potentiation, when it occurred, was accompanied by a decrease in the probability of occurrence of components with smaller peak amplitudes and an increase in the probability of occurrence of components with larger peak amplitudes. 7. These results are consistent with the suggestion that the magnitude of the synaptic potential generated at a single bouton does not vary from trial to trial (Jack, Redman & Wong, 1981a). Nor does the amplitude of this potential vary following a single conditioning stimulus or a tetanus. Post tetanic potentiation and facilitation result from a decrease in the probability of failure to release transmitter following the conditioning stimuli. PMID- 6279828 TI - Quantal analysis of a decremental response at hair cell-afferent fibre synapses in the goldfish sacculus. AB - 1. On application of a step decrement in the sound intensity, the amplitude of excitatory post-synaptic potentials (e.p.s.p.s) recorded intracellularly from large afferent auditory fibres in goldfish's sacculus showed a decremental response in which the amplitude of the e.p.s.p.s was temporarily reduced to a very low level, but soon returned to a new steady level appropriate to the decreased sound intensity. This response seems to underlie the temporary reduction in the rate of afferent discharge observed in the mammalian cochlea upon cessation of sound or upon reduction of its intensity 2. A statistical analysis revealed that reduction in the size of the mean quantal content (m) during the decremental response was associated with reduction in the size of binomial population (n), but not that of the probability (p) of any one of the available quanta actually being used. 3. The temporary reduction in the size of n during the decremental responses can be explained if it is assumed that replenishment to vacant release sites was channelled to high threshold sites and that the low threshold sites were bypassed. The mechanism underlying this special type of replenishment is discussed in relation to the ultrastructural features of presynaptic sites in the hair cell. PMID- 6279829 TI - Fluctuation analysis of neutral amino acid responses in cultured mouse spinal neurones. AB - 1. Intracellular recordings using the voltage-clamp technique were made at room temperature (24 +/- 1.5 degrees C) from mouse spinal and sensory neurones growing in dissociated cell culture. 2. Membrane current responses could be elicited by ionophoresis of the neutral amino acids, gamma-aminobutyric acid (GABA), beta alanine (BALA) and glycine to the cell body and processes of these neurones. 3. All membrane current responses were associated with increases in current fluctuations. Most of the analysis presented here was applied to responses generated at the cell body. 4. Many of the fluctuations in membrane current occurring during the responses could be interpreted as reflecting the kinetic behaviour of a single population of two-state Cl- ion-channels. 5. The properties of channels estimated during the desensitized phase of an amino acid-induced current response were not significantly different from those estimated during the peak of the response. 6. The properties of the amino acid-activated channels were relatively constant over the -40 to -90 mV range of membrane potential. 7. There was considerable variation in the estimated average conductance, gamma, and duration, tau, of the elementary events evoked by the neutral amino acids on spinal cord neurones. The properties of the elementary channel events activated by one of the amino acids were significantly different from those activated on the same neurones by either of the other amino acids. 8. In sensory neurones the average gamma and tau values for GABA-activated ion-channels were also determined and these values fell within the range of those for channels activated by GABA in spinal neurones. 9. The results indicate that different naturally occurring neutral amino acids activate channels with unique properties in cultured mouse spinal neurones. The relative charge transfer associated with these channels averages 1.00:0.74:0.32; GABA:glycine:beta-alanine. PMID- 6279830 TI - Stimulation by injected guanosine triphosphate of the sodium efflux in barnacle muscle fibres. AB - 1. A study has been made of the mechanism by which injected disodium GTP stimulates the ouabain-insensitive Na efflux in single muscle fibres from the barnacle, Balanus nubilus. 2. Injection of GTPNa2 causes a stimulatory response which is usually transitory and almost completely reversed by injecting MgCl2 (but not KCl). 3. Injected 5'-guanylylimidodiphosphate, Gpp(NH)p, mimics this action of GTP but the reversal seen with injected Mg2+ is less pronounced. 4. (i) Pre-treatment of these fibres with verapamil reduces the size of the stimulatory response to GTP and Gpp(NH)p. (ii) Pre-injection of protein kinase inhibitor (PKI) or regulatory subunits reduces the response as well. (iii) Pre-treatment with imipramine or trifluoperazine reduces the response to injected GTP; in combination with verapamil, a greater reduction in response is seen. 5. Injection of EDTA leads to a stimulatory response which is transitory. This response is largely abolished by verapamil. 6. Injection of cholera toxin leads to a sustained stimulatory rather than a transitory response. GTP or Gpp(NH)p when injected following peak stimulation by cholera toxin leads to a moderate sustained stimulation. 7. These results support the view that the stimulatory response to injected GTPNa2 is the result of activation of Ca2+ channels and of increased availability of GTPMg and that these two conditions bring about activation of adenylate cyclase and hence activation of cyclic AMP-protein kinase by newly formed cyclic AMP. PMID- 6279831 TI - Paracellular non-electrolyte permeation during fluid transport across rabbit gall bladder epithelium. AB - 1. Mucosa-to-serosa fluxes of seven polar non-electrolytes were determined during isotonic fluid transport across the unilateral rabbit gall-bladder preparation in an attempt to estimate the contribution of the paracellular pathway to the total transepithelial water flow.2. (3)H- and (14)C-labelled non-electrolyte tracers appeared in the transported fluid at fractions (f(n)) of their mucosal concentration which were inversely related to molecular size: ethanediol, 0.80; thiourea, 0.55; glycerol, 0.16; erythritol, 0.11; mannitol, 0.05; sucrose, 0.05; inulin, 0.02. The mean volume flow rate was 78 mul. cm(-2) hr(-1).3. While the fluxes of the larger molecules were probably due to diffusion through a small but unrestricted paracellular ;shunt' permeability, the high f(n) values obtained for the smaller molecules indicate the existence of a substantial paracellular permeability restricted to molecules smaller than erythritol.4. Upper limits to the transcellular ethanediol and thiourea permeabilities, estimated from the time constants of tracer efflux from preloaded epithelial cells, were too low to account for more than a very small fraction of the transepithelial fluxes observed in the unilateral preparation.5. Comparison of the f(n) values with the predictions of a hydrodynamic model of paracellular permeation suggests that in order to account for the large fluxes of ethanediol and thiourea, considerably more than one half of the transepithelial water flow must follow the paracellular pathway.6. Following a reduction of the mucosal osmolality to 110 m-osmole kg( 1), the apparent non-electrolyte permeability of the epithelium increased steadily over a period of 4 hr. This seems to reflect an increase in the shunt permeability rather than a change in the selectivity of the restricted permeability.7. It is concluded that during isotonic fluid transport the bulk of the transepithelial water flow crossing the epithelium passes through paracellular channels of approximately 3 A radius which are probably located in the intercellular junction. PMID- 6279832 TI - Evidence for the presence of potassium channels in the internode of frog myelinated nerve fibres. AB - 1. The central portion of single frog internodes under voltage-clamp conditions was acutely demyelinated with lysolecithin to resolve whether potassium channels are normally present only in the paranode (Chiu & Ritchie, 1980, 1981) or are in fact present throughout the entire internode of a myelinated nerve fibre. 2. The experiments were performed in the absence of nodes of Ranvier to ensure that all currents observed originated from the internode. 3. The demyelinating internodal segment showed a steady increase in the capacity and leakage for the first 40--50 min, after which a delayed outward current was revealed. This delayed current, which was blocked by TEA and caesium and exhibited a potassium-dependent reversal potential, seems to be a potassium current similar to that normally present in the frog node and in squid nerve. 4. The present method of acute demyelination revealed no detectable sodium currents in the internodal segments. 5. These results suggest that potassium channels, but virtually no sodium channels, are normally present throughout a frog internode covered by the myelin. PMID- 6279833 TI - On the quantal release of endogenous glutamate from the crayfish neuromuscular junction. AB - 1. The abdominal slow flexor muscle was isolated from the crayfish (Cambarus clarkii) and placed in 150 microliters. Harreveld solution. The concentrations of glutamate and aspartate in this solution were measured by mass fragmentography. 2. Application of black widow spider venom (BWSV) produced a marked increase in the frequency of miniature excitatory post-synaptic potentials (m.e.p.s.p.s). During the high frequency discharge of m.e.p.s.p.s, the glutamate content in the solution was significantly increased. There was an approximately linear relationship between the increase in the glutamate efflux produced by BWSV and the variance of the membrane potential fluctuation during high frequency discharge of m.e.p.s.p.s. 3. In most cases, the efflux of aspartate during control rest periods was smaller than that of glutamate. During the discharge of m.e.p.s.p.s produced by BWSV, the increase in the aspartate efflux was very small compared to glutamate. 4. Nerve stimulation caused a significant increase in the efflux of glutamate, but the change in the aspartate efflux was very small and not significant. 5. Application of methylene blue increased the frequency of m.e.p.s.p.s and glutamate efflux, but little, if any, increase was found in aspartate efflux. 6. It is concluded that glutamate is preferentially released from nerve terminals in a quantal fashion. PMID- 6279834 TI - [Contribution of ultrasonography to the study of retroperitoneal tumors in children (author's transl)]. AB - Ultrasonography investigations of retroperitoneal tumors in children were conducted in 25 cases: 11 nephroblastomas, 11 neuroblastomas, 2 ganglioneuromas, and 1 rhabdomyosarcoma. Ultrasonographic exploration of such tumors should include analysis of the mass: echostructure, limits, interfaces and dimensions, and analysis of its extension. The latter should determine the position in relation to the median line, the condition of the large vessels (inferior vena cava and aorta), the homolateral renal cavities and the contralateral kidney, and the presence or absence of glandular spread and hepatic metastases. The semiology of each of these elements is described and discussed, to determine their individual diagnostic value. PMID- 6279835 TI - Cellular aspects of the inhibitory actions of LH-RH on the ovary and testis. PMID- 6279836 TI - The role of oocyte maturation inhibitor in follicular regulation of oocyte maturation. PMID- 6279837 TI - Postpartum femoral neuropathy. AB - Two cases of postpartum femoral neuropathy occurred. The literature on this subject is reviewed, and possible etiologic mechanisms are discussed. Complete functional recovery is typical of the excellent prognosis of puerperal femoral neuropathy. The importance of recognizing this complication is to predict a favorable prognosis and thus eliminate anxiety for both patient and physician. PMID- 6279838 TI - Malignant fibrous histiocytoma treated by excision and repair with a free latissimus dorsi musculocutaneous flap. PMID- 6279839 TI - Amoebiasis: a review. PMID- 6279840 TI - Adenosine receptors: targets for future drugs. PMID- 6279842 TI - Synthesis and enzymic activity of various substituted pyrazolo[1,5-a]-1,3,5 triazines as adenosine cyclic 3',5'-phosphate phosphodiesterase inhibitors. AB - A series of various pyrazolo[1,5-a]-1,3,5-triazines have been prepared and studied as inhibitors of cAMP phosphodiesterase isolated from bovine brain, bovine heart, and rabbit lung. A number of compounds were found to be superior to theophylline. 2-Ethyl-7-phenylpyrazolo[1,5-a]-1,3,5-triazine (35) was found to be 97 times more potent than theophylline as an inhibitor of bovine brain PDE. 8 Bromo-2,4-dimethyl-7-phenylpyrazolo[1,5-a]-1,3,5-triazine (52) showed alpha lung = 40 compared to alpha heart = 3.0. Thus, various substituents could increase or decrease the inhibition relative to the type and source of tissue from which the PDE was isolated. The most active compound was 8-bromo-4-(diethylamino)-7 phenylpyrazolo[1,3-a]-1,3,5-triazine (25), which was 185 times more potent than theophylline as an inhibitor of PDE isolated from rabbit lung. The stepwise synthesis via ring-closure procedures of requisite pyrazole intermediates, followed by electrophilic substitution in the pyrazole ring and/or nucleophilic substitution in the 1,3,5-triazine moiety, resulted in the various pyrazolo[1,5 a]1,3,5-triazines listed in Tables I and II. Structure-activity relationships are reviewed. PMID- 6279841 TI - Synthesis and enzymic activity of 6-carbethoxy- and 6-ethoxy-3,7-disubstituted pyrazolo[1,5-a]pyrimidines and related derivatives as adenosine cyclic 3' ,5' phosphate phosphodiesterase inhibitors. AB - A number of 3,7-disubstituted 6-carbethoxypyrazolo [1,5-a] pyrimidines and 3,7 disubstituted 6-ethoxypyrazolo-[1,5-a]pyrimidines have been prepared and evaluated as adenosine cyclic 3',5'-phosphate (cAMP) phosphodiesterase (PDE) inhibitors vs. the low Km enzyme isolated from beef heart, rabbit lung, and kidney preparations. The results were found to be between 0.5 to 13 times as potent as theophylline as inhibitors of PDE, depending on the tissue source. A number of these PDE inhibitors exhibited significant physiological effects in different animal systems, suggesting it should be possible to obtain selective PDE inhibition in various tissues. Several of these heterocycles were found superior to adenosine in inhibiting ADP-induced platelet aggregation in vitro. PMID- 6279843 TI - Angiotensin-converting enzyme inhibitors: importance of the amide carbonyl of mercaptoacyl amino acids for hydrogen bonding to the enzyme. AB - A series of mercaptoacyl amino acids and related compounds was synthesized and evaluated for inhibition of angiotensin-converting enzyme (ACE) in order to determine the nature and importance of the putative interaction between ACE and the amide moiety of inhibitors such as captopril (3-mercapto-2-methylpropanoyl-L proline). It was concluded that the interaction involves a hydrogen bond from a donor site on ACE to the oxygen of the amide carbonyl. Compounds in which the amide moiety is replaced by other groups (ester, ketone, sulfonamide) capable of accepting a hydrogen bond are effective inhibitors, but compounds in which only the geometrical features of the amide are retained are ineffective inhibitors. The presence of an NH group is not necessary for effective inhibition. The activity of a series of mercaptoacyl cycloalkyl carboxylic acids parallels the activity of the isosteric series of mercaptoacyl imino acids. PMID- 6279844 TI - Use of adenine nucleotide derivatives to assess the potential of exo-active-site directed reagents as species- or isozyme-specific enzyme inactivators. 3. Synthesis of adenosine 5'-triphosphate derivatives with N6- or 8-substituents bearing iodoacetyl groups. AB - Several series of N6- or 8-substituted derivatives of adenosine 5'-triphosphate (ATP) were synthesized. N6-(omega-Aminoalkyl) derivatives of adenosine 5' monophosphate (AMP) were converted into their omega-N-carbobenzyloxy derivatives, and these were converted, via the 2',3'-O-carbonyl derivatives of their 5' phosphorimidazolidates, into the corresponding ATP derivatives. Hydrogenolytic removal of the carbobenzyloxy groups, followed by iodoacetylation of the omega amino groups with N-(iodoacetoxy)succinimide, gave N6-R-ATP, where R = (CH2)nNHCOCH2I (n = 2--8) or (CH2)nCON)CH3)(CH2)mN(CH3)CO(CH2)nNHCOCH2I (n = m = 3; n = 3, m = 4; n = 4, m = 3; n = m = 4). Condensation of N6-(omega-aminoalkyl) derivatives of AMP with N-hydroxysuccinimide esters of omega-[N (carbobenzyloxy)amino] carboxylic acids gave N6-(CH2)nNHCO(CH2)mNH-Cbz derivatives of AMP which, upon conversion to the corresponding derivatives of ATP, followed by removal of the carbobenzyloxy group and iodoacetylation, as described above, gave N6-(CH2)nNHCO(CH2)mNHCOCH2I-ATP derivatives (n = 3, m = 5 or 6; n = 4, m = 5; n = 6, m = 1--6). The same sequence of reactions starting with N6-[omega-(methylamino)alkyl] derivatives of N6-CH3-AMP gave N6-CH3, N6 (CH2)nH(CH3)CO(CH2)mNHCOCH2I derivatives of ATP (n = 4, m = 3, 5 or 6; n = 6, m = 5 or 6). Reaction of alpha, omega-diaminoalkanes with 8-Br-ATP gave 8-NH(CH2)nNH2 derivatives of ATP, which upon iodoacetylation gave 8-NH(CH2)nNHCOCH2I derivatives of ATP (n = 2, 4, 6, or 8). Substrate and inhibitor properties indicated that the ATP derivatives are potential exco-ATP-site-directed inactivators of hexokinases, adenylate kinases, and pyruvate kinases. PMID- 6279845 TI - Use of adenine nucleotide derivatives to assess the potential of exo-active-site directed reagents as species- or isozyme-specific enzyme inactivators. 4. Interactions of adenosine 5'-triphosphate derivatives with adenylate kinases from Escherichia coli and rat tissues. AB - Adenosine 5'-triphosphate (ATP) derivatives of the types N6-R-ATP [R = (CH2)nHNCOCH2I, (CH2)nNHCO-(CH2)mHNCOCH2I, or (CH2)nCON(Me)(CH2)mN(Me)CO(CH2)nNHCOCH2I], N6-Me-N6-R-ATP [R = (CH2)nN (Me)CO(CH2)mNHCOCH2I], and 8-R-ATP [R = NH(CH2)nNHCOCH2I] with 5--19 spacer atoms between N6 or C-8 and iodine have been evaluated as substrates, reversible inhibitors, and inactivators of adenylate kinase (AK). With Escherichia coli AK, the derivatives were noncompetitive inhibitors, Ki = 4.7--7.3 mM, with little affinity for the ATP site, and N6-(CH2)nNHCOCH2[-ATP (n = 5 or 6) effected progressive inhibitions that were not ATP site directed. With rat muscle AK (M AK), some compounds had slight affinity for the ATP site as evidenced by weak substrate activity with as much as 8 spacer atoms, but all compounds tested were weak noncompetitive inhibitors; Ki = 6--12 mM vs. ATP. The ATP derivatives, notably N6-(CH2)8NHCOCH2I-ATP, mediated a progressive inhibition of M-AK, which was abolished by substitution of hydrogen for the iodine and thus presumably involves alkylation of the enzyme. The inhibition appeared not to be ATP site directed because kinetic analysis indicated a random bimolecular enzyme-inhibitor reaction and because N6-(CH2)8NHCOCH2I-AMP and its adenosine counterpart, which have relatively low affinity for the ATP site, were more effective than N6 (CH2)8NHCOCH2I-ATP. The ATP derivatives were substrates (KM = 0.4--1.6 mM) and/or competitive inhibitors (Ki = 0.3--6.2 mM) vs. ATP of rat isozymes AK II or III. Exposure of AK II or III for 6 h, 22 degrees C, at pH 7.6 to 10 mM levels of the 1:1 Mg complexes of 25 of the ATP derivatives led in no case to progressive enzyme inhibition, suggesting the absence near the ATP sites of nucleophilic groups suitably positioned for alkylation. PMID- 6279846 TI - 2-(Alkylthio)-1,2,4-triazolo[1,5-a]pyrimidines as adenosine cyclic 3',5' monophosphate phosphodiesterase inhibitors with potential as new cardiovascular agents. AB - A series of new 2-(alkylthio)-5,7-disubstituted-1,2,4-triazolo[1,5-a]pyrimidines have been prepared as inhibitors of cAMP phosphodiesterase from various tissues. These derivatives were prepared via ring closure of various requisite 3-amino 1,2,4-triazole intermediates. 2-(Benzylthio)-5-methyl-7-(dimethylamino)-1,2,4 triazolo[1,5-a]pyrimidine (15a) is 6.3 times as potent as theophylline in inhibiting cAMP PDE isolated from rabbit heart. Treatment of dogs intravenously with 5 (mg/kg)/h of 15a gave a cardiac output increase of 69%, which was largely sustained for a 2-h period after administration of drug had ceased. There was no significant increase in heart rate upon administration of 15a. Related studies with 5,7-di-n-propyl-2-(benzylthio)-1,2,4-triazolo[1,5-a]pyrimidine (22a) in five dogs showed a 31.5% increase in cardiac output with an increase in stroke volume of 34.4% with no increase in heart rate. The specificity of action of these PDE inhibitors could be due to selective binding at a certain cAMP PDE site in the cardiovascular system. Several of these compounds are candidates for further studies with a view to clinical evaluation. PMID- 6279847 TI - Chemical synthesis and molecular pharmacology of hydroxylated 1-(1 phenylcyclohexyl-piperidine derivatives. AB - The following monohydroxy derivatives of 1-(1-phenylcyclohexyl)piperidine (phencyclidine, PCP) were synthesized: o-, m-, and p-phenols of PCP, 1-(1 phenylcyclohexyl)-4-piperidinol, and two stereoisomeric pairs of 3-phenyl-3-(1 piperidinyl)cyclohexanol and 4-phenyl-4-(1-piperidinyl)cyclohexanol. Inhibition of specific binding of tritiated PCP, morphine, or quinuclidinyl benzylate (QNB) in rat brain homogenates was measured for these compounds. Inhibition of PCP binding for selected compounds correlated with mouse rotarod assay activity. The most characteristic effects of hydroxylation of PCP on the cyclohexyl, piperidine, or phenyl moieties are the following: (i) it generally decreases its activity in inhibiting [3H]PCP binding by a factor of 10 to 80; (ii) it does not produce a large variation in the affinity for the morphine receptor; (iii) it produces a considerable decrease of the affinity for the muscarinic receptor. An important exception to these general observations was the metaphenolic derivative of PCP. This PCP derivative has an affinity for the [3H]PCP binding sites that is 8 times higher than that of PCP itself; its affinity for the muscarinic receptor is only twice lower than that of PCP, but its affinity for the morphine receptor is 430 times higher than that of PCP and only one order of magnitude lower than that of morphine itself. PMID- 6279848 TI - A microtiter assay for cell-mediated cytotoxicity to cytomegalovirus antigens. AB - The cell-mediated immune response is important to host defense against viruses, yet techniques with which to measure the contribution of lymphocytes (particularly K cells) to anti-viral cytotoxic reactions are not well established. We describe a new procedure for measuring lymphocyte-mediated antibody-dependent immunity to cytomegalovirus. Central to the assay is the use of 51Cr-containing red blood cells labeled with purified viral antigen, which can be easily quantitated and readily stored for at least 72 hours. High sensitivity was observed, as sera with a 1: 512 complement fixation titer could be diluted 10,000-fold without extinguishing the detectable cytotoxic response. A set of conditions (target/effector ratio; dilution of serum; source and age of target cells; and incubation medium and time) was defined that optimized sensitivity, maintained a low level of background release, yet utilized small amounts of reagents. A kinetics study revealed that 80% of specific release of radioisotope occurred in the first 4 hours. Investigation of the epidemiology, diagnosis, and management of disease caused by CMV may be greatly aided by this new method for quantitating specific cell-mediated anti-viral immune reactivity. PMID- 6279849 TI - An evaluation under code of new techniques for the detection of cytomegalovirus antibodies: sensitivity of assays and importance of immune complexes. AB - Given the morbidity and occasional mortality associated with cytomegalovirus infection and the requirement for good seroepidemiologic tools, assays of high sensitivity and reliability are needed for detection of cytomegalovirus-specific antibody. We report the successful application of two sensitive techniques, antibody-dependent cellular cytotoxicity (ADCC) and the enzyme-linked immunosorbent assay (ELISA) for the measurement of these antibodies and document their favorable comparison with conventional techniques, complement fixation (CF) and indirect hemagglutination (IHA). A coded panel of 48 sera from patients with culture-proven cytomegalovirus infection and disease controls was tested in a controlled fashion by all four procedures. Among 18 CF-positive sera, specific antibody was detected by IHA, ADCC and ELISA in dilutions 100 to 1000-fold higher than measurable by the CF test. Ten of 15 sera, seronegative by CF, were nevertheless found to be cytomegalovirus antibody-positive when assessed by the other procedures. Further, the importance of circulating antigen-antibody complexes upon ADCC test results was documented. Eight sera, known to be antibody positive by other techniques but with an unexpectedly low (or negligible) capacity to induced ADCC, were found to be immune complex-positive by the Raji cell assay. Ultracentrifugation, useful in precipitating antigen-antibody complexes, enhanced ADCC activity in five of six sera known to be complex positive. It is suggested that the simultaneous application of multiple antibody detection systems, particularly the newly-developed ones (ADCC, ELISA) will provide both earlier diagnostic information as well as a more comprehensive understanding of the human host's immune response to infection with cytomegalovirus. PMID- 6279850 TI - Experimental murine hepatitis and inducible suppressor cell function. AB - We have investigated the changes in inducible suppressor activity in spleen cells of young BALB/c mice infected with A-59 strain of murine hepatitis virus. The peak histological and biochemical damage occurred four days after intraperitoneal inoculation of the virus. Before that, on day 2, there was a transient loss of suppressor activity inducible in splenic cells by in vitro incubation with the mitogen Con A. Sequential changes in the proliferative response of spleen cells to Con A also occurred, but did not account for the loss of inducible suppressor activity. This variation in activity of Con A-stimulated suppressor cells suggests an immunoregulatory role for these cells during experimental viral hepatitis. PMID- 6279851 TI - Interaction between a synthetic analogue of vasopressin and human peripheral blood lymphocytes. AB - The interaction between 1-desamino-8-D-arginine vasopressin (DDAVP) and human peripheral blood lymphocytes was investigated in vitro, using DDAVP concentrations ranging between 1 pg and 10 microgram per ml. DDAVP did not display any mitogenic activity in the absence of phytohaemagglutinin (PHA). Neither did DDAVP have any significant effect on the PHA-stimulated incorporation of 3H-thymidine into DNA, provided that optimally mitogenic concentrations of PHA were present. However, DDAVP either enhanced or inhibited lymphocyte transformation initiated by suboptimally mitogenic concentrations of PHA: the direction of the response depended on the magnitude of the transformation response induced by the PHA alone. Experiments employing 125I-DDAVP demonstrated that DDAVP bound reversibly to untransformed lymphocytes, and that at 20 degrees C, the amount bound increased rapidly to reach a plateau value after 60 minutes. The binding of DDAVP was inhibited in the presence of Pitressin or arginine vasopressin suggesting that human peripheral blood lymphocytes have surface receptors for the vasopressins which also react with DDAVP. PMID- 6279852 TI - Loss of the short-lived suppressive function of peripheral leukocytes in feline retrovirus-infected cats. AB - Normal feline peripheral blood mononuclear cells demonstrated increased phytomitogen blast transformation after a 24 hr delay in mitogen addition as compared to 0 hr mitogenesis. Increased stimulation after 24 hr which is not due to enhancing factors accumulating in the culture medium has been attributed to a short-lived suppressor cell which loses its effect after 24 hr in culture. This short-lived suppressive phenomenon was studied in normal specific-pathogen-free cats and compared to experimental animals infected with retrovirus (FeLV). Experimental cats demonstrated a subsequent loss of this function following acquisition of a persistent viremia. PMID- 6279853 TI - Spontaneous generalized Herpesvirus hominis infection of a lowland gorilla (Gorilla gorilla gorilla). AB - A 13-day-old lowland gorilla died from a generalized herpesvirus infection shortly after the onset of clinical signs. The pathologic-anatomical findings were compatible with those described for generalized herpes simplex infection in the human neonate. Electron microscopic examination of lung tissue revealed the presence of herpesvirus which was identified with the fluorescent antibody technique as Herpes simplex virus type I. Tests with related sera of the herpes group (varicella, herpesvirus-B) revealed no specific immunoflourescence. PMID- 6279854 TI - Malignant tumours in Tupaia (tree shrew). PMID- 6279855 TI - Stoichiometry of proton movements coupled to ATP synthesis driven by a pH gradient in Streptococcus lactis. AB - An electrochemical potential difference for H+ was established across the plasma membrane of the anaerobe Streptococcus lactis by addition of sulfuric acid to cells suspended in potassium phosphate at pH 8 along with valinomycin or permeant anions. Subsequent acidification of the cell was measured by the distribution of salicyclic acid. A comparison between cells treated or untreated with the inhibitor N,N'-dicyclohexylcarbodiimide was used to reveal that portion of net proton entry attributable to a direct coupling between H+ inflow and synthesis of ATP catalyzed by the reversible proton-translocating ATPase of this microorganism. When the imposed electrochemical proton gradient was below 180-190 mV, proton entry was at the rate expected of passive flux, for both control cells and cells treated with the ATPase inhibitor, However, at higher driving force acidification of control cells was markedly accelerated, coincident with ATP synthesis, while acidification of cells treated with the inhibitor continued at the rate characteristic of passive inflow. This observed threshold (180-190 mV) was identified as the reversal potential for this H+ "pump". Parallel measurements showed that the free energy of hydrolysis for ATP in these washed cells was 8.4 kcal/mole (370mV). The comparison between the reversal (threshold) potential and the free energy of hydrolysis for ATP indicates a stoichiometry of 2 H+/ATP for the coupling of proton movements to ATP formation in bacteria. PMID- 6279856 TI - Identification of a structural hairpin in the filamentous chimeric phage M13Gori1. PMID- 6279857 TI - Crystallization and preliminary x-ray diffraction data for the collagenase of Hypoderma lineatum larvae. PMID- 6279858 TI - Integration and transcription of group C human adenovirus sequences in the DNA of five lines of transformed rat cells. PMID- 6279859 TI - Sequence rearrangements between mitochondrial DNAs of Torulopsis glabrata and Kloeckera africana identified by hybridization with six polypeptide encoding regions from Saccharomyces cerevisiae mitochondrial DNA. PMID- 6279860 TI - Spin-label study of actin-myosin-nucleotide interactions in contracting glycerinated muscle fibers. PMID- 6279861 TI - Sequential folding of a messenger RNA molecule. PMID- 6279862 TI - Methylated and unmethylated ribosomal RNA genes in the mouse. PMID- 6279863 TI - The nature of the complexes formed between the int protein and DNA. PMID- 6279864 TI - Site-specificity of the chromosomal insertion of Staphylococcus aureus transposon Tn554. PMID- 6279865 TI - Kinetics and mechanism of the association of the bacteriophage T4 gene 32 (helix destabilizing) protein with single-stranded nucleic acids. Evidence for protein translocation. PMID- 6279866 TI - Site-specific recombination Xis-independent excisive recombination of bacteriophage lambda. PMID- 6279867 TI - Conformation change of cytochrome c. I. Ferrocytochrome c structure refined at 1.5 A resolution. PMID- 6279868 TI - Conformation change of cytochrome c. II. Ferricytochrome c refinement at 1.8 A and comparison with the ferrocytochrome structure. PMID- 6279869 TI - Cloning of F DNA fragments carrying the origin of transfer oriT and the fertility inhibition gene finP. PMID- 6279871 TI - Gene amplification in methotrexate-resistant mouse cells. II. Rearrangement and amplification of non-dihydrofolate reductase gene sequences accompany chromosomal changes. PMID- 6279870 TI - Gene amplification in methotrexate-resistant mouse cells. I. DNA rearrangement accompanies dihydrofolate reductase gene amplification in a T-cell lymphoma. PMID- 6279872 TI - Gene amplification in methotrexate-resistant mouse cells. III. Interrelationships between chromosome changes and DNA sequence amplification or loss. PMID- 6279873 TI - Short intervening sequences close to the 5' ends of the three Drosophila larval serum protein 1 genes. PMID- 6279875 TI - Isolation and mapping of ribosomal RNA genes of Caulobacter crescentus. PMID- 6279874 TI - Sequence and expression of the discoidin I gene family in Dictyostelium discoideum. PMID- 6279876 TI - Crystallographic structure of Rhodospirillum molischianum ferricytochrome c' at 2.5 A resolution. PMID- 6279877 TI - Complexes formed between the restriction endonuclease EcoK and heteroduplex DNA. PMID- 6279878 TI - Late messenger RNA production by viable simian virus 40 mutants with deletions in the leader region. PMID- 6279879 TI - Cytochrome c4 from Pseudomonas aeruginosa. PMID- 6279880 TI - Histological effects and lead concentrations in tissues of adult male ringed turtle doves that ingested lead shot. AB - Adult male ringed turtle doves (Streptopelia risoria) were administered 0 or 4 110 mg lead shot and/or were exposed to cold temperatures (6 +/- 1 degrees C). Five of the seven doves that ingested shot and were exposed to cold died. Doves that ingested shot and were not exposed to cld (21 +/- 1 degrees C) had no mortality after 9 d, but several birds had seizures. Tissues were examined microscopically for lesion. Doves ingesting shot consistently had Pb intranuclear inclusion bodies in cells of the proximal convoluted tubules, except if death occurred 48 h after shot ingestion. In such cases as in cold-exposed, Pb-treated birds), intracytoplasmic inclusions were detected. Hemosiderin loading in kupffer cells and rarely in hepatocytes was observed in doves ingesting Pb whether they were exposed to a normal temperature or to cold. it appeared that ingestion of shot could abruptly disturb spermatogenesis in ringed turtle doves. The seminiferious tubules were often devoid of spermatozoa in doves ingesting Pb. PMID- 6279882 TI - [Acute toxicological studies of captopril in rats and mice]. AB - The acute toxicity of captopril, a potential inhibitor of angiotensin converting enzyme, was studied in Sprague-Dawley rats and ICR mice. In the oral administration, a decrease of spontaneous motor activity, lacrimation, salivation and decline of body temperature was noted in the rats and mice. In the mice, tarry stool was also noted. The LD50 was estimated as: 4249 mg/kg in male mice, 5050 mg/kg in female mice, 4336 mg/kg in male rats and 4245 mg/kg in female rats, In the dead animals, a hemorrhagic erosion or ulcer was recognized in the glandular stomach. An intravenous captopril in mice, caused immediate death by dyspnea in some animals within 3 minutes, but delayed death was also occurred within 24 hours showing a decrease of spontaneous motor activity and decline of body temperature. The LD50 was estimated as 3154 mg/kg in male mice and 3225 mg/kg in female mice. A single intravenous administration of captopril in dose of 1600 mg/kg did not cause any death in the rats of both sexes. The mice of both sexes well tolerated a single subcutaneous administration of captopril in a dose of 2400 mg/kg. No death occurred in rats of both sexes received subcutaneously captopril in a dose of 1200 mg/kg. At an injection site, a necrotic ulcer was noted in the skin of rats and mice which received subcutaneously captopril in a dose of 1600 mg/kg or more and 1200 mg/kg, respectively. PMID- 6279881 TI - Acute and chronic effects of pentobarbital in relation to postsynaptic GABA receptors: a study with muscimol. AB - Muscimol, a GABA agonist, enhanced pentobarbital sleeping time in a dose dependent manner. The GABA antagonists such as bicuculline and picrotoxin, and the CNS stimulant such as pentylenetetrazol, inhibited pentobarbital sleeping time; however, all except picrotoxin produced less than 35% maximum inhibition. Picrotoxin, and agent which blocks the chloride ionophore of GABA-receptor complex, exhibited a parallel dose-response curve with respect to muscimol. Chronic administration of pentobarbital by pellet implantation induced tolerance as evidenced by decreased sleeping time; the tolerance receded gradually upon abrupt withdrawal. Muscimol enhanced pentobarbital sleeping time both in tolerant and withdrawal mice. Na+-independent GABA-receptor binding, using [3H]muscimol as a ligand, was increased after acute and chronic pentobarbital administration; withdrawal of the pentobarbital reversed the increase in receptor population. None of the treatments altered the affinity of [3H]muscimol binding. These results support the contention that pentobarbital (a) directly acts on the postsynaptic chloride ionophore and (b) augments GABA-mediated postsynaptic effects. The functional significance of the increase in GABA receptor population after pentobarbital treatment is unclear. PMID- 6279883 TI - [One month studies on the subacute toxicity of captopril in rats]. AB - Subacute toxicity of captopril by daily oral administration at dose levels of 10 mg/kg, 30 mg/kg, 100 mg/kg, 300 mg/kg, 900 mg/kg and 2700 mg/kg for one month was studied in Sprague-Dawley rats. In the 2700 mg/kg group, 13 of 18 males and 17 of 18 females died with marked emaciation and abdominal distension. In the 900 mg/kg group, 1 of 18 males and 3 of 18 females died also during the administration period. In dead animals, a marked dilatation of gastrointestinal tract was noted showing multiple hemorrhagic erosions and/or ulcers in the glandular stomach. The remainder of these groups exhibited polydipsia and polyuria during the dosage period. In the 300 mg/kg and 100 mg/kg groups, all animals survived throughout the entire experimental period showing polydipsia and polyuria. In the 30 mg/kg and 10 mg/kg groups, all animals survived also throughout the entire dosage period without showing any toxic sign. Regarding plasma analysis, the BUN and creatinine concentration was significantly elevated in the group of 100 mg/kg or more. In the hematological examination, there was a decrease of erythrocyte counts, hemoglobin contents and hematocrit values in the group of 300 mg/kg or more. Pathological examinations revealed a marked thickening of the wall in afferent arterioles and interlobular arteries of the kidney in association with hypertrophy and hyperplasia of juxtaglomerular cells in 100 mg/kg, 300 mg/kg, 900 mg/kg and 2700 mg/kg groups. In these groups, multiple hemorrhagic erosions with or without ulcer were also noted in the glandular stomach. In the spleen, a slight increase of extramedullary hematopoiesis and hemosiderosis was noted in the 100 mg/kg, 300 mg/kg, 900 mg/kg and 2700 mg/kg groups, in which an increase of erythropoietic elements was also noted in the bone marrow. From these results, the maximum nontoxic dose was estimated as about 30 mg/kg/day by oral administration of captopril in the rats. PMID- 6279884 TI - [Twelve month studies on the chronic toxicity of captopril in rats]. AB - The chronic administration of captopril to Sprague-Dawley rats was performed under the barrier system by feeding ad libitum with mixed diet in various concentrations of captopril with 3 months recovery period. The number of animals was 180 female and 180 male including 5 groups of control, 30, 100, 300 and 900 mg/kg/day. The maximum nontoxic dose was estimated as about 30 mg/kg/day for male but a little more than this for female rats. Body weight increase was significantly reduced in male but for the first 3 months in female rats. No death was ascribed to the toxic effect of captopril. Polydipsia and polyuria in male, and the significant increase in values of BUN and inorganic phosphate in both sexes were observed. The reduction in erythrocyte count, values of hemoglobin and hematocrit, hemosiderosis in reticulum cells of the spleen and Kupffer cells in the liver and the increase of erythropoieses indicated hemolytic anemia. Heart weight reduced while kidney weight increased. Pathological examination revealed hypertrophia and hyperplasia of JG cells and thickening of walls of afferent arterioles with hyperplasia of vascular smooth muscle cells and increase of collagen fibers. Thickening of walls extended to walls of the interlobular arteries which remained after withdrawal of captopril for 3 months though JG granules attenuated. The age-related increases of incidences of proteinuria and myocardial fibrosis were attenuated dose-dependently which are probably due to hypotension induced by captopril. PMID- 6279885 TI - [Three month subacute toxicity of captopril in beagle dogs]. AB - Thirty four beagle dogs, male and female were orally given 10, 30, 100 and 200 mg/kg/day of captopril, an angiotensin converting enzyme inhibitor, for 3 months followed by a recovery test for 4 weeks. One of 4 female dogs which were treated with the highest dose of 200 mg/kg/day throughout the experimental period died of bronchial pneumonia. Hypersalivation and occasionally vomiting was observed in dogs treated with 100 and 200 mg/kg/day. Skin eruption such as erythema and papules was observed mostly at the ventral surface of the neck, chest and upper abdomen in dogs in these two experimental groups. Histological examination of the lesion revealed cellular infiltration with edema and expansion in the dermis and slight hyperkeratosis with parakeratosis and acanthosis. Changes in erythrocyte counts, hematocrit values and hemoglobin contents during the course of administration were variable among dogs but these were obvious in animals treated with higher doses. An increase in erythropoiesis of the bone marrow, extramedullary hematopoiesis and slight hemosiderosis in liver and spleen were revealed by histological examination. Above histological observations suggest that captopril may cause hemolysis. Hypertrophy and hyperplasia of juxtaglomerular cells with increased number of JG granules were shown in the highest dosage group even 4 weeks after suspension of captopril administration. A distinct plasma renin activity supported the morphological changes. From the results of three months administration of captopril to beagle dogs, the maximum non-toxic dose may be around 10 mg/kg/day and toxic dose 100 mg/kg/day. PMID- 6279886 TI - Improved cadaver allograft survival in transfused recipients who remain serologically negative for cytomegalovirus. AB - Between April 1976 and October 1979, 100 consecutive cadaver renal transplants were done. Before transplantation 48 recipients were seronegative and 52 were seropositive for cytomegalovirus. After transplantation there were 20 primary and 38 secondary cytomegalovirus infections. The development of post-transplant cytomegalovirus infection, with or without overt symptoms, had no effect on graft survival. The 1-year graft survival was significantly better (p less than 0.05) in high transfused (more than 5 units) recipients (70 per cent) compared to nontransfused recipients (36 per cent). The beneficial effect of transfusions was not diminished in patients with positive post-transplant cytomegalovirus serology. Of the transfused recipients those who remained serologically negative for cytomegalovirus pre-transplant had significantly better 1-year graft survival (78 per cent) than those who were cytomegalovirus positive before transplantation (58 per cent) (p less than 0.05). The improved graft survival in patients who remain cytomegalovirus seronegative after multiple blood transfusions may be a manifestation of unresponsiveness to immunologic as well as virogenetic stimulation. PMID- 6279887 TI - Adult Wilms tumor: effect of combined therapy on survival. AB - Thirty-one adults with Wilms tumor were reported to the National Wilms Tumor Study from 1968 to 1979. Treatment and survival data for these patients were analyzed and compared to similar information derived from children enrolled in the first National Wilms Tumor Study. The ages of the 31 adults ranged from 17 to 63 years (mean 29 years). All but 3 patients had surgical resection or excision of tumor, 7 did not receive postoperative irradiation and all but 1 had chemotherapy. Actinomycin D and vincristine were the drugs used most commonly, 26 of the 31 patients receiving both agents. Advanced disease at diagnosis (6 stage III and 9 stage IV versus 9 stage I and 5 stage II--in 2 cases stage was not known) was found more often than in children in whom stages III and IV disease made up 27 per cent of the first National Wilms Tumor Study population. The 3 year actuarial survival rate for the 31 adults was 24 per cent: 48 per cent for stages I and II disease and 11 per cent for Stage IV disease. Comparable data for children in the first National Wilms Tumor Study, adjusted for stage, were 74, 87 and 53 per cent, respectively. It is concluded that adults with Wilms tumor treated as were these have a worse prognosis than children managed according to the first National Wilms Tumor Study regimen. However, those adults in this series who were treated aggressively, that is surgical excision, postoperative irradiation and multi-agent chemotherapy, appeared to have fared better than adults treated in the pre-chemotherapy era. It is concluded that aggressive therapy should be given to all adults with Wilms tumor irrespective of stage. PMID- 6279888 TI - Homozygous cystinuria--evaluation of 35 patients. PMID- 6279889 TI - Assessing ureteral wound healing by pressure flow technique. PMID- 6279891 TI - Prevalence of antibodies to parainfluenza-3 virus in various wildlife species and indigenous cattle sharing the same habitats in Kenya. PMID- 6279890 TI - Beta-endorphin levels in blood from selected Alaskan mammals. AB - Blood samples were analyzed for beta-endorphin from 43 non-torpid black bear (Ursus americanus), 8 torpid black bear, 3 non-torpid brown bear (Ursus arctos), 14 moose (Alces alces), 6 mountain goats (Oreamnus americanus) and 30 Steller sea lions (Eumetopias jubatus). Beta-endorphin levels were detected in all species sampled and there were no significant differences in levels among non-torpid black bear, brown bear and sea lions. Also, no differences were detected between moose and mountain goats, but all other comparisons were significantly different (P less than 0.001). Torpid black bear had higher levels than all other groups. Moose and mountain goats had the lowest levels. The possibility of beta-endorphin influencing behavior and physiology of mammals is discussed. PMID- 6279892 TI - Presence of hepatitis B surface antigen before primary hepatocellular carcinoma. AB - In a study of 7,498 men of Japanese ancestry in Hawaii, 18 incident cases of primary hepatocellular carcinoma were identified after the follow-up period of 13 years. Sixteen of the 18 patients had stored serum specimens available for analysis. Ten of the 16 had positive serum test results for hepatitis B surface antigen in their original specimens. None of 48 matched controls subjects, whose stored serum specimens were also analyzed, had similar positive test results. This finding indicates that hepatitis B surface antigenemia preceded primary hepatocellular carcinoma in many patients. PMID- 6279893 TI - Carrier status in hepatitis B: a modern scourge with hope. PMID- 6279894 TI - Leukemia patients to be given varicella vaccine. PMID- 6279895 TI - Birth defects and vaginal spermicides. AB - In a cohort study of 50, 282 pregnancies recruited between 1958 and 1965, there were 462 gravidae who used nonmercurial spermicides (mostly nonoxynol-9 (95% confidence limits, 0.6 to 1.6). There were also 889 women who used phenylmercuric acetate (no longer available as a spermicide); the corresponding rate ratio was 0.9 (0.6 to 1.3). Limb reduction deformities, neoplasms, Down's syndrome, and hypospadias did not occur in excess in children exposed to spermicides. PMID- 6279896 TI - Transfusion-related cytomegalovirus infection following noncardiac surgery. AB - Clinically evident cytomegalovirus (CMV) infections developed in five patients after noncardiac surgery. Each of them had received several blood transfusions and had evidence of primary CMV infection. These five cases were identified in a single eight-month period, suggesting that this syndrome must be occurring with greater frequency than is commonly appreciated. Three of five patients had undergone splenectomy, and review of the literature for documented cases of posttransfusion CMV mononucleosis following non-cardiac-bypass surgery disclosed an additional nine patients, five of whom had had splenectomies. This striking association suggests the possibility that the spleen plays a role in controlling the incidence or clinical manifestations of posttransfusion CMV infections. Alternatively, this association may only reflect the frequent requirement for many blood transfusions in this type of surgery. PMID- 6279897 TI - Herpes simplex antigen in immune complexes of patients with erythema multiforme: presence following recurrent herpes simplex infection. AB - Serum samples from patients with erythema multiforme (EM) were examined for the presence of herpes simplex virus (HSV) antigen in immune complexes using the Raji cell radioimmunoassay. Raji-cell-bound immune complexes from eight of 12 patients with EM after HSV infection and four of four patients with EM of uncertain cause had increased anti-HSV binding, while patients with EM after drug exposure and patients with recurrent HSV infections without EM had binding in the same range as controls. Viral cultures for HSV of immune complexes eluted from Raji cells were negative. Sucrose density gradient ultracentrifugation studies of serum samples of patients with EM after HSV infection showed HSV antigen in large molecular weight fractions. The HLA typing of lymphocytes from 16 patients with EM after HSV infection was not different from that of controls. Immune complexes composed of antibody and HSV antigen are present in serum samples of patients with EM after HSV infection and some cases of EM of uncertain cause and may mediate the pathogenesis of these disorders. PMID- 6279898 TI - Clostridium perfringens empyema unresponsive to penicillin. PMID- 6279899 TI - [Effect of visceral sensation on the dorsal horn unit activity in cat spinal cord. (2) Effects of changes of intravesical pressure of gall bladder on the unit activity and modification of the activity with thiamylal sodium under the influence of intravesical pressure (author's transl)]. PMID- 6279900 TI - [Effects of intrathecal beta-endorphin and morphine on endocrine response during halothane anesthesia and surgery in man (author's transl)]. PMID- 6279901 TI - [Effects of flunitrazepam-pentazocine-N2O anesthesia and surgery on plasma catecholamines, ACTH, cortisol and ADH levels in man (author's transl)]. PMID- 6279902 TI - [Clinical evaluation of ceftizoxime in the pediatric infections (author's transl)]. AB - Ceftizoxime (FK 749, CZX) was evaluated in 24 children with a suspicion of bacterial infection. Of the 17 confirmed bacterial infections, 16 were shown to be effective (effective rate, 94.1%). The diagnosis included acute pharyngitis (2), pneumonia (6), staphylococcal empyema (1), cervical purulent lymphadenitis (2), acute enterocolitis (2), acute pyelonephritis (1), SSSS (1) and suspected septicemia (2). The etiological pathogens recovered were Streptococcus anginosus (1), Streptococcus pneumoniae (1), Staphylococcus aureus (2), Haemophilus influenzae (3), enteropathogenic Escherichia coli (1) etc. A case of suspected Pseudomonas aeruginosa septicemia was not effectively treated with CZX. The serum half-life of CZX was 1.36 hours after intravenous bolus infection. A cerebrospinal fluid level of CZX was 6.2 mcg/ml 1 hour after intravenous bolus injection of 1 g (23.8 mg/kg) in a child with inflamed meninges. No severe adverse reaction was encountered with the CZX therapy. The data suggest that CZX is an excellent candidate for the first choice parenteral antibiotic in the pediatric infections. PMID- 6279904 TI - [Pharmacokinetics and clinical effects of ceftizoxime in pediatric field (author's transl)]. AB - Pharmacokinetics and clinical effects of ceftizoxime (CZX), a new cephalosporin antibiotic, were investigated and following results were obtained. 1) Ceftizoxime was given by intravenous injection or drip infusion for 1 hour at a single dose of 30 mg/kg. After intravenous injection, the mean peak serum level of 3 children was 95.9 mcg/ml at 15 minutes and half-life time was 1.18 hours. After 1 hour drip infusion, the mean peak serum level of 3 children was 79.5 mcg/ml at the end of infusion and half-life time was 1.20 hours. The urinary level was high and the mean urinary recovery rate was 69.6% and 63.4% up to 6 hours after intravenous injection and 1 hour drip infusion, respectively. 2) CZX was administered in dose of 39--76 mg/kg to 7 pediatric patients (4 cases of purulent meningitis, 2 of septicemia with purulent meningitis, and 1 of aseptic meningitis) by a single intravenous injection. In patients with purulent meningitis, passage into the cerebrospinal fluid was relatively as good as 30% of serum level at the same time in the presence of remarkable signs of inflammation, but poor in cases of mild inflammation or aseptic meningitis. 3) Cerebral puncture fluid level in 1 patient with cerebral abscess was as good as 65.5% of serum level at the same time. 4) CZX was given to 28 cases of respiratory tract infection, 1 of tonsillitis with otitis media, 6 of scarlet fever, 1 each of maxillary sinusitis and bacterial endocarditis, 6 of purulent meningitis, 2 of septicemia, 5 of septicemia suspected, 2 of septicemia with purulent meningitis, 1 each of osteomyelitis, typhoid fever, peritonitis and biliary tract infection, 16 of urinary tract infection, 14 of skin and soft tissue infection, and 1 of external otitis, totaling 87 cases. The mean daily dose of 101.6 mg/kg was administered for an average of 10 days mainly by intravenous injection 4 times daily. Clinical results obtained were excellent in 34 cases, and good in 46. Bacteriological effectiveness rate was 100%. As for side effects, fever, fever with rash, fever with cough and diarrhea appeared in 1 each case out of 182 cases including 95 drop out cases. As for laboratory findings, eosinophilia, thrombocytopenia, elevation of GOT, that of GOT with GPT, and that of GOT with LDH appeared in 10, 2, 2, 3 and 1 cases, respectively. PMID- 6279903 TI - [Clinical studies on ceftizoxime in pediatric field (author's transl)]. AB - Clinical studies on ceftizoxime, a new cephalosporin, were carried out in our department. The following results were obtained. 1. Antibacterial activity. Antibacterial activity of ceftizoxime against 7 strains of E. coli, 6 strains of Klebsiella, 6 strains of H. influenzae, 7 strains of E. cloacae and 10 strains of S. aureus, recently isolated from patients, was compared with that of cefotiam, cefmetazole and cefazolin. Ceftizoxime was more active than the other antibiotics against E. coli, Klebsiella, H. influenzae and E. cloacae, but less active against S. aureus. 2. Urinary excretion. Urinary excretion was measured in 2 cases with normal renal function after dosing with 750 mg (35 mg/kg) and 350 mg (17 mg/kg) of ceftizoxime by intravenous injections. Urinary recovery rates within 6 hours were 97% and 82% respectively. 3. Clinical study. Eighteen children with the following bacterial infections were treated with ceftizoxime; respiratory tract infection (13), acute otitis media (1), acute intervertebral chondritis and tonsillitis (1), chronic cystitis (1), subcutaneous abscess (1) and chronic bacteremia (1). The dosage was 69--147 mg/kg q.i.d. by intravenous injection. The duration of administration was from 3 to 32 days. The clinical results were excellent in 4 cases, good in 13 cases and fair in 1 case of chronic bacteremia. The overall effectiveness rate was 94%. Slight elevation of GPT in 1 case and leukopenia (neutropenia) in 1 case were observed, but returned to the normal range immediately after discontinuation of dosing. It is considered that ceftizoxime is one of the useful first choice antibiotics used for children with bacterial infections. PMID- 6279905 TI - [Clinical studies on ceftizoxime in children (author's transl)]. PMID- 6279906 TI - [Fundamental and clinical studies of ceftizoxime in pediatric infections (author's transl)]. AB - Experimental and clinical trials were carried out with ceftizoxime in pediatric infections. Results were as follows. 1. The antibacterial activity of ceftizoxime against clinically isolated organisms was determined. Ceftizoxime was more active than cefazolin, cefotiam and cefmetazole against K. pneumoniae, E. coli and H. influenzae. 2. The serum concentrations of ceftizoxime following intravenous injection of 10 mg/kg were 16.2, 10.1, 6.2, 1.8 micrograms/ml at 30, 60, 120, 240 minutes after injection, respectively. Ceftizoxime was excreted with the rate of 97.2% in the 6-hour urine after injection. 3. Twenty-one patients comprising 5 with urinary tract infection, 16 with respiratory tract infection were received ceftizoxime at the doses ranging from 32 to 109 mg/kg divided 3 or 4 times a day. The results were excellent in 8 and good in 13 patients. The rate of satisfactory clinical response was 100%. 4. Slight and transient elevation of GPT was observed in 1 patient. Other side effects were not observed in any of these 21 patients. PMID- 6279907 TI - [Laboratory and clinical studies of ceftizoxime in pediatrics (author's transl)]. AB - Laboratory and clinical studies were performed as follows on ceftizoxime (CZX), a new cephalosporin antibiotic. 1. Susceptibility of clinically isolated bacteria to CZX and cefotiam (CTM) or sulbenicillin (SBPC). Antibacterial activities of CZX and CTM were compared against S. aureus, E. coli, K. pneumoniae, E. cloacae, H. influenzae and E. aerogenes; CZX was compared with SBPC against Ps. aeruginosa. CZX and CTM were nearly equal in activity against S. aureus, but CZX was found to be more active than CTM by 1--10 tubes against E. coli, K. pneumoniae, E. cloacae, H. influenzae and E. aerogenes. Against Ps. aeruginosa, CZX and SBPC were nearly equal in activity. 2. Serum concentration and urinary recovery. Serum concentrations of CZX were measured in 6 patients given CZX for prophylactic purpose during cardiac catheterization. In 2 patients given 20 mg/kg of CZX intravenously, the average serum concentration was 38.9 micrograms/ml at 30 minutes after intravenous bolus injection. In 3 patients given 10 mg/kg of CZX by intravenous drip infusion, the peak average serum concentration was 28.1 micrograms/ml at the end of infusion. Urinary recovery in 2 patients tested was 81.1% and 92.5% until 6--7 hours after intravenous bolus injection. 3. Clinical efficacy. CZX was given intravenously to 24 patients in doses of 30--111 mg/kg (57.1 mg/kg on an average) t.i.d. or q.i.d. for 3--16 days (5.5 days on an average): 1 with lacunar tonsillitis, 4 with acute bronchitis, 12 with pneumonia, 2 with enterocolitis, 2 with soft tissue infection, 2 with lymphadenitis and 1 with UTI. The overall efficacy rate was 95.8%, i.e., efficacy was excellent in 10 (41.7%), good in 13 (54.2%), and poor in 1 (4.2%). Bacteriological efficacy was excellent, i.e. 21 of the 23 strains disappeared. One patient had mild and transient diarrhea, but no other laboratory abnormalities were observed during treatment. The above results suggest that CZX is 1 of the most useful antibiotics for treating pediatric infections, especially due to Gram negative bacteria. PMID- 6279909 TI - [Ceftizoxime preparations]. PMID- 6279908 TI - [Pharmacokinetics and clinical evaluation of ceftizoxime (author's transl)]. AB - Pharmacokinetics of ceftizoxime (CZX), a new cephalosporin antibiotic, was investigated in 9 children with normal renal and hepatic function. In addition, the clinical effect of CZX was evaluated in 26 pediatric patients with various infections. In 4 of the 9 children with normal renal and hepatic function, intravenous bolus injection of CZX in a dose of 20 mg/kg yielded a mean peak serum level of 36.5 micrograms/ml at 1/2 hour after infusion, and mean serum levels of 12.5 micrograms/ml at 2 hours and 6.0 micrograms/ml at 4 hours after infusion. The biological half-lives of CZX were estimated to be 1.25--2.55 hours. In another child, serum levels of CZX at 1/2, 2 and 4 hours after intravenous bolus injection in a dose of 10 mg/kg were 19.60, 5.96 and 2.06 micrograms/ml, respectively. The clear difference in dose response between 20 mg/kg and 10 mg/kg reflected the doubled dose levels. In the remaining 4 children, drip infusion of CZX in a dose of 20 mg/kg (1 child 17 mg/kg) over 0.5--1.5 hours yielded peak serum levels at the end of infusion. The biological half-lives of CZX were estimated to be 0.95--1.50 hours. About 80% of CZX was excreted in the urine within 6 hours after infusion in the 4 children tested. Twenty-six pediatric patients with various infections were treated with CZX intravenous doses of 20 mg/kg to 118 mg/kg b.i.d.--q.i.d. for 3--14 days. Of the 12 patients with acute bronchitis and pneumonia, 5 showed excellent response, 6 good and 1 fair response. Of the 5 patients with urinary tract infection, 4 showed excellent response and 1 good response. One patient each with colitis, tonsillitis and facial cellulitis, pharyngitis showed excellent response and 1 patient each with purulent thyroiditis and gluteal abscess showed good response. The single patients with sepsis showed excellent response. One patient each with pyothorax, purulent arthritis and cerebral abscess showed poor response. Overall effectiveness rate was 84.6%. although 22 of all 26 patients treated had serious underlying diseases such as APL, AML. A mild increase in GOT and GPT was observed in 1 patient during treatment with CZX, and the values returned to normal after discontinuation of the drug. These results suggest that ceftizoxime is 1 of the most important antibiotics for treating a wide range of infections in children as well as in adults. PMID- 6279910 TI - [Clinical studies with cefmenoxime in the otorhinolaryngeal field (author's transl)]. AB - We conducted a therapeutic trial with cefmenoxime in the field of otorhinolaryngology on a total of 14 subjects including 7 cases of otitis media, 5 cases of peritonsillar abscess, 1 case of chronic sinusitis and 1 case of congenital aural fistula. 1. The effective rate for peritonsillar abscess was extremely high being judged 'markedly effective' in 3 cases and 'effective' in 2 cases. 2. The effective rate for otitis media was also remarkably high; it was judged effective in 85.7% of the cases. It is thought that this is probably due to the fact that therapy and supervision were carried out on inhospital patients. 3. The 1 case of chronic sinusitis and the 1 case of congenital aural fistula were each judged 'effective' and 'markedly effective'. The only side effect observed was a slight case of skin eruption in 1 of the patients. PMID- 6279911 TI - [Concentrations of cefmenoxime and cefotiam in the human bone marrow blood (author's transl)]. AB - In order to examine the venous blood and bone marrow blood concentrations of cefmenoxime (CMX) and cefotiam (CTM) in man, 1 g of CMX or CTM was administered by one shot intravenous injection prior to surgery of the hip joint. At 30, 60, 120 and 180 minutes after the administration, venous blood and bone marrow blood were collected from each of 3 patients and the concentration assayed by the agar well method. Both CMX and CTM showed excellent distribution to bone marrow blood with peak levels of 50.1 micrograms/ml and 50.9 micrograms/ml on the average 30 minutes after administration. The levels of CMX in bone marrow blood exceeded those in the venous blood at 60 minutes and thereafter and CTM at 30 minutes of administration and thereafter. It is therefore considered that both CMX and CTM appear to be useful drugs for the prophylaxis and treatment of bone marrow infection, e.g. osteomyelitis. PMID- 6279912 TI - [Fundamental and clinical studies in pediatric field on ceftizoxime (author's transl)]. AB - Fundamental and clinical studies in the pediatric field on ceftizoxime were carried out, and the following results were obtained. 1. In 4 children age from 3 years to 5 years, the serum concentrations and urinary excretion of ceftizoxime in a dose of 20 mg/kg by intravenous drip infusion over 60 minutes were measured. The peak serum levels were 22.0--84.0 microgram/ml (mean 45.0 microgram/ml) at the end of infusion. The mean serum levels after the end of infusion were 16.9 microgram/ml at 30 minutes, 12.1 microgram/ml at 1 hour, 6.2 microgram/ml at 2 hours, 1.6 microgram/ml at 4 hours and 0.6 microgram/ml at 6 hours, with mean serum half-life (T 1/2) of 1.03 hours, mean urinary recovery rate was 64.9% up to 6 hours. 2. Concentrations of the drug in the cerebrospinal fluid in 1 patient with purulent meningitis at 30 minutes after an intravenous drip infusion of about 33.3 mg/kg were 0.2 to 1.5 microgram/ml, which were 8 to 60 times higher than the MICs of the causative organisms. 3. Ceftizoxime was administered to 38 children with pneumonia, bronchitis, Salmonella enteritis, purulent meningitis, etc. in the daily dose of 44--200 mg/kg for 3--19 days. Clinical response was excellent in 24, good in 12, poor in 1 and unknown in 1. The drug was proved to be very effective in 1 case of purulent meningitis due to H. influenzae. As for side effect, eruption was observed in only 1 case. PMID- 6279913 TI - [Fundamental and clinical studies on ceftizoxime in pediatric field (author's transl)]. AB - Fundamental and clinical studies of ceftizoxime, a new cephalosporin antibiotic, in children led to the following results. 1. Ceftizoxime compared favorably with cefazolin (CEZ) and cefmetazole (CMZ) for in vitro activity against clinically isolated strains of Staphylococcus aureus (31 strains), Escherichia coli (29), Klebsiella pneumoniae (30) and Pseudomonas aeruginosa (16). While somewhat less active against S. aureus than CEZ and CMZ, ceftizoxime was far more active than these 2 cephalosporin antibiotics against the test strains of E. coli and K. pneumoniae, which included strains resistant to the 2 drugs. Ceftizoxime was not particularly active against Ps. aeruginosa, but this seeming disadvantage was offset by the absolute ineffectiveness of the 2 reference drugs on this obstinate organism. 2. The time course of mean serum ceftizoxime levels in 3 pediatric patients of 5--10 years old given a single intravenous dose of 20 mg/kg was as follows: 45.4 micrograms/ml at 15 minutes, 40.4 micrograms/ml at 30 minutes, 22.1 micrograms/ml at 1 hour, 10.4 micrograms/ml at 2 hours, 2.9 micrograms/ml at 4 hours and 0.9 microgram/ml at 6 hours. The mean serum half life was 1.12 hours. The mean urinary levels of ceftizoxime at serial 2-hour collection intervals were as follows: 2,477 micrograms/ml for 1--2 hours, 1,235 micrograms/ml for 2--4 hours and 462 micrograms/ml for 4--6 hours. The mean urinary recovery up to 6 hours was 61.0%. 3. The clinical response of 28 children with infection to ceftizoxime treatment was 'excellent' in 22 children, 'good' in 4, and 'poor' in 2. These children comprised 11 with acute pneumonia, 3 with acute bronchitis, 4 with acute pyelonephritis, 2 each with acute purulent arthritis and acute enterocolitis, and 1 each with acute purulent tonsillitis, acute purulent lymphadenitis, furunculosis, subcutaneous abscess, subdural abscess and sepsis. The overall rate of effectiveness was 92.9%. Successfully eradicated strains in the bacteriological sense consisted of 4 strains each of H. influenzae and E. coli, 1 strain each of P. morganii, S. pneumoniae and S. pyogenes, 1 of the 2 strains of S. enteritidis, and 1 of the 3 strains of S. aureus. The overall rate of bacteriological effectiveness was 81.3%. No clinical side effects were observed. Changes in laboratory test findings included slightly and transiently elevated GOT and GPT in 1 child and GOT alone in another child. PMID- 6279914 TI - [Laboratory and clinical studies on ceftizoxime (author's transl)]. AB - The authors have carried out the laboratory and clinical studies of ceftizoxime (CZX), and obtained the following results. 1. The antibacterial activities of CZX were measured by plate dilution method against clinical isolates of S. aureus, E. coli, K. pneumoniae and P. aeruginosa. CZX inhibited the growth of S. aureus at concentrations less than 12.5 micrograms/ml, and the peak of sensitivity distribution was obtained at 3.13 micrograms/ml with an inoculum size of 10(6) cells/ml. And the peak sensitivity distribution of E. coli and K. pneumoniae were obtained at less than 0.1 microgram/ml and that of P. aeruginosa was obtained at 6.25 micrograms/ml. 2. Phagocytosis was determined by Quie's method. Phagocytosis of E. coli and K. pneumoniae by human polymorphonuclear neutrophil was more enhanced in the presence of 1 MIC and 1/2 MIC of CZX than of CEZ at 4 and 6 hours after incubation. 3. As for pharmacokinetic study, CZX was given by intravenous injection and drip infusion for 1 hour at a single dose of 10 mg/kg and 30 mg/kg. After intravenous injection of 10 mg/kg and 30 mg/kg of CZX, the mean peak serum levels were 19.1 +/- 3.4 micrograms/ml and 69.1 micrograms/ml at 30 minutes, and half-life times were 1.20 hours and 1.35 hours, respectively. After 1 hour drip infusion of 10 mg/kg and 30 mg/kg of CZX, the mean peak serum levels were 28.8 +/ 3.6 micrograms/ml and 60.9 +/- 5.9 micrograms/ml at the end of infusion, and half-life times were 1.40 hours and 1.77 hours, respectively. The mean urinary excretion rates were between 75.3% and 101% up to 6 hours after intravenous injection and drip infusion. 4. CZX was given to 4 cases with tonsillitis, 3 with pneumonia, 1 with enteritis, 4 with U.T.I., totaling 21 cases. A daily dose of CZX between 350 mg and 2,000 mg was given for 3 to 5 days. Clinical results obtained were good in all cases. No side effects and abnormal laboratory findings were observed. PMID- 6279915 TI - [Clinical experience with ceftizoxime in the pediatric field (author's transl)]. AB - Ceftizoxime (CZX) was given intravenously to 28 children with the following acute bacterial infections; 15 cases of bronchopneumonia, 4 cases of urinary tract infection, 3 cases of purulent meningitis, 2 cases of acute tonsillitis, each 1 case of purulent cervical lymphadenitis and acute tonsillitis, orbital cellulitis, abscess of eyelid, and peritonitis. Out of 28 patients, good clinical responses were obtained in 26 patients, and bacteriological effectiveness was seen in all 14 cases. Side effects with CZX were observed in 2 cases; 1 case was drug fever, and the other case was eosinophilia. From the above clinical results, CZX is useful antibiotic for treating the pediatric patients with various kinds of bacterial infections. PMID- 6279917 TI - [Fundamental and clinical studies on ceftizoxime (author's transl)]. AB - Ceftizoxime was tried in children with various infections and the following results were obtained. 1) Serum levels and urinary recovery of ceftizoxime were studied in 6 patients aged from 6 to 10 years. After intravenous bolus injection of 20 mg/kg, the mean serum concentrations were 113.9 micrograms/ml, 47.6 micrograms/ml, 31.8 micrograms/ml, 18.5 micrograms/ml, 6.3 micrograms/ml and 2.1 micrograms/ml at 5 minutes, 30 minutes, 1 hour, 2 hours, 4 hours and 6 hours, respectively. The average urinary recovery rates of ceftizoxime were 55.1%, 90.9% and 98.3% at 2 hours, 4 hours and 6 hours after the administration, respectively. 2) The therapeutic efficacy was excellent in 11, good in 9 and poor in 2 patients, the efficacy rate being 91%. 3) As for the side effects, elevation of S GOT and S-GPT, and drug fever were observed in 1 case, but disappeared soon after discontinuation of the therapy. PMID- 6279916 TI - [Laboratory and clinical studies on ceftizoxime in the field of pediatrics (author's transl)]. AB - Ceftizoxime, a new cephalosporin preparation, was evaluated for its antibacterial activity, absorption, excretion and clinical effectiveness, and the following results were obtained. The minimum inhibitory concentrations (MICs) of ceftizoxime against 211 clinical isolates were determined in comparison with those of cefazolin, cefmetazole, cefotiam and 6059 S. Against S. pyogenes (50 strains), ceftizoxime was 1 tube inferior to cefazolin inoculum size of 10(8) cells/ml, but was 2--3 tubes superior to cefmetazole and 6059-S. Against E. coli (50 strains), ceftizoxime and 6059-S were significantly more active than the other drugs. The susceptibility pattern of Klebsiella sp. (50 strains) to ceftizoxime was similar to that to cefotiam and 6059-S. Against Proteus sp. (50 strains), cefotiam and 6059-S were more active than the other drugs. Ceftizoxime was intermediate in activity, and cefazolin was the least active. Against H. influenzae (11 strains), ceftizoxime was the most active, with concentrations of 0.1 mcg/ml required to inhibit 100% of strains with an inoculum size of 10(8) cells/ml and 10(6) cells/ml. A dose of ceftizoxime 10 mg/kg or 20 mg/kg was administered to 15 patients aged from 5 years to 12 years, and serum levels and urinary excretion of the drug were measured. Intravenous bolus injection of the drug in dose of 10 mg/kg and 20 mg/kg yielded mean serum levels of 26.6 mcg/ml and 55.7 mcg/ml at 30 minutes, respectively. The serum levels of the drug, thereafter, declined gradually but still remained 1.3 mcg/ml and 2.7 mcg/ml at 6 hours. The serum half-lives (T 1/2) were estimated to be 1.17 hours in dose of 10 mg/kg and 1.31 hours in dose of 20 mg/kg. When a dose of 20 mg/kg was infused over a period of 30 minutes, the serum levels attained the peak of 72.4 mcg/ml to 82.4 mcg/ml (mean 79.4 mcg/ml) at the end of infusion. The levels, thereafter, tapered to mean levels of 45.3 mcg/ml at 30 minutes, 24.7 mcg/ml at 1 1/2 hours, and 3.6 mcg/ml at 5 1/2 hours, with a T 1/2 of 1.22 hours. Meanwhile, when the same dose was infused over 1 hour, the serum levels attained the peak of 59.4 mcg/ml to 68.5 mcg/ml (mean 64.2 mcg/ml). The mean serum levels after the end of infusion were 41.3 mcg/ml at 30 minutes, 21.6 mcg/ml at 1 hour and 1.9 mcg/ml at 5 hours, with a T 1/2 of 0.97 hours. Urinary recovery of the drug was 69.2% to 79.9% after intravenous injection and 62.3% to 79.9% after drip infusion, most of the given drug was excreted in the first 2 hours after administration. In our clinical study, 27 children with moderate or severe infections (12 cases of bronchopneumonia or bronchitis, 5 of pyelonephritis, 3 of purulent meningitis, etc.) were treated with ceftizoxime at the daily dose of 30--309 mg/kg for 3--23 days. Clinical response was excellent in 10, good in 9, fair in 5 and poor in 3. The drug was proved to be very effective against infections due to H. influenzae K. pneumoniae, E. coli and S. aureus. No serious side effects were observed in any case. PMID- 6279918 TI - Effect of high plasma free fatty acids on the free radical formation of myocardial mitochondria isolated from ischemic dog hearts. AB - Effects of high plasma free fatty acids (FFA) on the free radical formation of myocardial mitochondria, isolated from normal and ischemic dog hearts, were studied by electron spin resonance (ESR) spectrometry. Free radical concentrations in state 4 respiration were used for the evaluation of the function in the mitochondria in this study. High plasma FFA levels were induced either by intravenous injection of Intralipid and heparin, or by infusion of norepinephrine. Ischemic hearts were induced by inserting a Cournand's 7F catheter into the left coronary artery under fluoroscopic control. Exogenous high plasma FFA induced by Intralipid and heparin caused the decrease of free radicals in state 4 respiration in the mitochondria isolated from normal and ischemic dog hearts. Endogenous high FFA induced by continuous infusion of norepinephrine also caused the decrease of free radicals. On the other hand, nicotinic acid prevented the decrease of free radicals as well as the rise of plasma FFA by the norepinephrine infusion. These results suggest that high plasma FFA itself, whether it may be exogenous or endogenous, may impair the oxidative phosphorylation of the mitochondria isolated from normal and ischemic hearts. PMID- 6279919 TI - [Hepatitis caused by known viruses (other than A and B viruses)]. PMID- 6279920 TI - [Liver neoplasm accompanied by frequent attack of hypoglycemia--a case report and 91 other cases reported in Japan]. PMID- 6279921 TI - [TRH, its function and receptor]. PMID- 6279923 TI - [Recent findings on ACTH]. PMID- 6279922 TI - [LH-RH and analogues]. PMID- 6279924 TI - [Endorphins and opiate receptors]. PMID- 6279926 TI - [Biological functions and receptors for prolactin]. PMID- 6279925 TI - [Gonadotropin receptors]. PMID- 6279927 TI - [Vasopressin and receptor]. PMID- 6279928 TI - [PTH, thyrocalcitonin (author's transl)]. PMID- 6279929 TI - [Thyroid hormone receptor]. PMID- 6279930 TI - [Determination and action mechanism of mineralocorticoids]. PMID- 6279931 TI - [Recent findings on glucagon receptors and isolation of glicentin]. PMID- 6279932 TI - [Recent findings on catecholamine receptors]. PMID- 6279933 TI - [Angiotensin II and its receptors]. PMID- 6279934 TI - [Prostaglandin and its receptors]. PMID- 6279935 TI - [Congenital ACTH unresponsiveness]. PMID- 6279936 TI - [Electron microscopic autoradiography of diffusible compounds]. PMID- 6279937 TI - [Malignant fibrous histiocytoma--an autopsy case]. PMID- 6279938 TI - [Studies on 5'-nucleotidase. I. An automated micromethod based on the coupled enzymatic reactions and the serum activities in healthy subjects (author's transl)]. PMID- 6279939 TI - [Serological investigation of Epstein-Barr virus antibodies and heterophile antibodies of childhood in Kumamoto Prefecture (author's transl)]. PMID- 6279940 TI - Acute demyelination in mice inoculated intraspinally with mouse hepatitis virus, JHM strain. AB - After intracerebral or intralumbar inoculation of mouse hepatitis virus, JHM strain, into weanling mice, viral growth and lesions in the central nervous system were comparatively studied for 5 days postinoculation. In the intralumbar group the virus titer of the spinal cord exceeded that of the brain until 24 hr postinoculation, and declined later. In the intracerebral group the brain virus titer was always higher, and the cord titer attained the same level as that of intralumbar group at 72 hr. Demyelinating lesions in the spinal cord appeared 48 hr after intralumbar inoculation, that is, 36 hr earlier than after intracerebral inoculation. Virus-specific fluorescence was detected within glia cells in the cord white matter 48 hr intralumbar inoculation. Electron microscopy revealed a number of virions within oligodendroglia cells, suggesting that the acute phase demyelination might be due to viral growth within this type of cells. PMID- 6279942 TI - [Fundamental study on radioisotopic lymphography of the head and neck (author's transl)]. PMID- 6279941 TI - Electron microscopy of massive cerebrocortical necrosis in suckling rats inoculated intracerebrally with mouse hepatitis virus, A59 strain. AB - Cerebrocortical lesions produced in suckling rats by intracerebral inoculation with MHV-A59 was studied by electron microscopy. Many cytoplasmic vesicles and irregular patterns of the nuclear membrane were observed in infected neurons at 48 h post-inoculation (p.i.). The abnormally electron-lucent or -dense neurons were seen at 72 h p.i., and in the formers, many virions were detected between the outer and inner nuclear membranes as well as in the dilated endoplasmic reticulum cisternae. Both reticular and tubular inclusion bodies appeared in the cytoplasm and some virions seemed to be associated with an end of the latter. Many virions were also seen in the intercellular spaces. At 96 h p.i. many virions were present not only in the cytoplasm of degenerated neurons but also in astrocytes appearing in the spongy lesions. There was also degeneration of oligodendrocytes while having no virions. In rats which survived the acute phase of infection and were killed at 8 days or 14 days p.i., many macrophages and fibroblasts were accumulated in the thickened leptomeninges and submeningeal area. At 22 days p.i., atrophied electron-dense neurons having no virions were scattered throughout the cerebral cortex. PMID- 6279943 TI - Sympathetic nervous systems in chronic hypoxic states from pulmonary tuberculosis: a clinical study on plasma norepinephrine and cyclic AMP levels. AB - To evaluate the sympathetic nervous activity in chronic hypoxic states, the plasma concentrations of norepinephrine (NE), cyclic adenosine 3', 5' monophosphate (cyclic AMP), cyclic guanosine 3', 5'-monophosphate (cyclic GMP) and dopamine-beta-hydroxylase (DBH) enzyme activity were measured in 32 patients with pulmonary tuberculosis and 30 normal volunteers. Moreover, the peripheral arterial-venous difference of concentration of these substance, arterial blood gas levels and spirometric data were studied in these patients. The results of this study demonstrated that the plasma NE and cyclic AMP concentrations were directly related to the degree of hypoxemia and some of the spirometric parameters in these patients. It is concluded that sympathetic nervous activity was augmented in chronic hypoxic states, especially with chronic respiratory failure. PMID- 6279944 TI - Vitamin D resistant hypophosphatemic osteomalacia associated with osteosarcoma of the mandible: report of a case. AB - A case of vitamin D resistant hypophosphatemic osteomalacia associated with osteosarcoma of the mandible is presented. The patient complained of lumbar, knee and foot pain and muscle weakness of two years' duration. Serum phosphorous was 1.0-1.6 mg/dl, tubular reabsorption of phosphorus was 47 to 58%, TmPO4/GFR was o.7-1.2 mg/dl. Aminoaciduria was noted. Bone biopsy confirmed the diagnosis of osteomalacia. He partially responded to the treatment with 1 alpha()H) D3 and sodium phosphate. After removal of sarcoma of the mandible, symptoms remitted and pertinent laboratory data became normal except serum alkaline phosphatase for more than one year without treatment. It is suggested that an impaired response of the tubule and bone to active vitamin D3, caused in some way by the osteosarcoma might be one of the causes of osteomalacia in this case. PMID- 6279945 TI - Brain damage after deep hypothermia in dogs. AB - Histopathological and succinic acid dehydrogenase (SAD) activity changes, and the electron spin resonance (ESR) were examined in the brains of dogs exposed to various hypothermic conditions. A simple hypothermic cardiac arrest of 5 min duration occurred in the group subjected to 28 degrees C, and normal SAD activity was noted in dogs exposed 1-hour extracorporeal circulation under normothermia. In cases of a hypothermia below 28 degrees C, a decrease in enzyme activity, marked histopathological changes and the disappearance of the ESR signal were observed. In dogs killed immediately after induction of 20 degrees C hypothermia, the histopathological changes were primarily ischemic in nature, while in dogs killed at 1-12 month intervals after the experiments, irreversible gliosis was dominant. Our findings suggest that, to avoid brain damage under conditions of deep hypothermic circulatory arrest, the critical range is between 26-28 degrees C for rectal, and approximately 28-30 degrees C for brain temperature. PMID- 6279946 TI - [Pulmonary emphysema (author's transl)]. PMID- 6279947 TI - Effects of three avian respiratory viruses on pathogenicity in tracheal organ cultures. PMID- 6279948 TI - [Serogical studies on an epidemic prevalence of swine type influenza virus among swine and human beings (author's transl)]. PMID- 6279949 TI - Isolation of picornavirus from horses associated with Getah virus infection. PMID- 6279950 TI - Aftercare instruction: wound care. From the Patient Teaching Committee, Department of Emergency Nursing, Hennepin County Medical Center. PMID- 6279951 TI - Characterization of a monoclonal antibody reactive with a subset of human breast tumors. AB - Fusion of spleen cells, from mice immunized against the human hormone-dependent breast cancer cell line ZR-75-1, with murine myeloma cells resulted in the establishment of a hybridoma cell line, HY59-H10 (H59). The purified monoclonal IgM produced by the hybridoma reacts with the most differentiated human breast tumor cell lines but not with cells derived from normal breast secretions or with numerous other malignant cell lines. Of 106 biopsy specimens examined, H59 bound to 54% of malignant breast specimens, to 83% of fibroadenomas, and to 82% of specimens containing fibrocystic disease. PMID- 6279952 TI - Moloney murine sarcoma virus tumors in CBA/J mice: chemopreventive and chemotherapeutic actions of supplemental beta-carotene. AB - Decreased tumor frequency, increased latent period, and increased rate of tumor regression were observed in male inbred CBA/J mice fed supplemental beta-carotene before and/or after they were inoculated with the Moloney sarcoma virus. When beta-carotene feeding was begun after tumors were already present, it markedly increased the rate of tumor regression. beta-Carotene minimized the virus-induced thymus gland involution that accompanies tumor growth, and this action on the thymus gland was believed to underlie part of beta-carotene's antitumor activity. The basal diet, a standard commercial mouse chow containing more vitamin A than the National Research Council recommends as a daily allowance for rodents, supported normal growth, reproduction, and longevity of normal mice. The work reported here is the first demonstration of the antitumor action of beta-carotene in mice inoculated with an oncogenic virus. PMID- 6279954 TI - [Radionuclide methods of evaluating ischemic heart disease]. PMID- 6279953 TI - Malignant fibrous histiocytomas induced by 4-(hydroxyamino)quinoline 1-oxide in rats. AB - Subcutaneous and bone malignant fibrous histiocytomas (MFH) were induced in high incidence in rats by 4-(hydroxyamino)-quinoline 1-oxide (4-HAQO). Subcutaneous MFH were induced locally by repeated weekly injections of 1 mg 4-HAQO/rat for 4 weeks. Between 16 and 48 weeks after the final treatment, 13/15 (87%) male noninbred Wistar rats developed tumors. The histologic subtypes of these tumors were as follows: 11 fibrous, 1 myxoid, and 1 giant cell. Bone MFH were induced between 18 and 25 weeks by implantation of 4-HAQO (8 mg/rat) into the bone marrow of the tibia in 12/14 (86%) male noninbred Fischer 344 rats. The histologic subtypes of these tumors were as follows: 6 fibrous, 4 myxoid, and 2 giant cell. Morphologically, these MFH appeared similar to human MFH. PMID- 6279955 TI - [99mTc-pyrophosphate scintigraphic indices during the bicycle ergometry test in myocardial infarct in the early phase of healing]. AB - Scintigraphy with 99mTc-pyrophosphate during bicycle ergometry has been undertaken in 32 patients with myocardial infarction in the early phase of its healing. There were 3 scintigraphic investigations: one during bicycle ergometry and two control ones - before and the day after the test. The determination of changes in the degree of 99mTc-pyrophosphate accumulation under the influence of the exercise in patients with myocardial infarction in the early phase of its healing permitted the authors to detect several variants in the time course of scintigraphic values. Drop or rise of concentration of the labelled pyrophosphate at the height of the bicycle ergometry test compared to the data obtained prior to it are apparently due to changed bloodflow in the focus of lesion during exercise. Changes in the character and intensity of accumulation of the radiodrug after bicycle ergometry can be explained in the majority of cases by increase of metabolic disorders in the myocardium, supervening during the exercise, and in some patients, possibly, to sustained decrease of bloodflow in the affected area of the cardiac muscle. PMID- 6279956 TI - [Level of cyclic nucleotides in the urine of patients with acute pneumonia]. PMID- 6279957 TI - [Levels of catecholamines and cyclic nucleotides in infection]. PMID- 6279958 TI - [Differential diagnostic potentials in myocardial ischemia and necrosis using 99mTc-pyrophosphate]. PMID- 6279959 TI - [Hormone level in the blood during food loading in peptic ulcer and chronic gastritis]. PMID- 6279960 TI - Host factors in Coxsackievirus B4-induced pancreopathy. AB - The diabetogenic potential of the human isolate, Coxsackievirus B4 (CB4) (Edwards) was studied in three inbred mice strains, SWR/J, DBA/2, and C57BL/6. The mice were infected with this agent and evaluated for mortality, pancreatic histopathology, and glucose tolerance. Results showed that the mortality induced by CB4 in these inbred strains differed considerably. There was no evidence of a correlation between virus-induced mortality and virus-induced pancreopathy. Although CB4 (Edwards) was most lethal to C57BL/6 mice, based on the infecting 50 per cent lethal dose (LD50), this mouse strain developed no pancreatic pathology. The most severe pancreopathy, i.e., acinar necrosis with acute interstitial inflammation and islet atrophy, was observed in SWR/J mice, which had an intermediate susceptibility to virus-induced mortality. DBA/2 mice, which displayed the lowest susceptibility to virus-induced lethality, showed less pancreatic pathology (i.e., acute and chronic interstitial inflammation) than SWR/J mice. IN SWR/J mice, virus-mediated alteration in glucose homeostasis was expressed by an increase in glucose tolerance 7 and 21 days after infection. In contrast, C57BL/6 mice showed a tendency toward chemical diabetes at 21 days postinfection. This study suggests that CB4-induced mortality and pancreatic pathology are independent parameters and do not necessarily determine the glucose tolerance of a given host genotype. PMID- 6279961 TI - A critical review of biological methods for the detection of fungal toxins in foods and foodstuffs. PMID- 6279962 TI - The interaction of corticosteroids and naloxone in canine hemorrhagic shock. PMID- 6279963 TI - The catechol estrogens. PMID- 6279965 TI - Effect of steroids on viral infection of human breast cancer cell lines by adenovirus 5. PMID- 6279964 TI - Studies on the mode of action of calciferol-XXXV. Comparison of the biochemical properties and ligand specificities of receptors and antibodies for 1,25 dihydroxyvitamin D3. PMID- 6279967 TI - Hormone binding of estradiol-17 beta receptor: evidence for its regulation by cytoplasmic phosphorylation and nuclear dephosphorylation. Prevention of dephosphorylation by antiestrogens. PMID- 6279966 TI - In vitro study of frog (Rana ridibunda Pallas) interrenal function by use of a simplified perifusion system-VI. Inhibition of aldosterone biosynthesis by SC 14266. PMID- 6279968 TI - Growth inhibitory effects of thyroid hormones on androgen-dependent mammary tumor cells. PMID- 6279969 TI - ACTH induction of the maturation of ACTH-sensitive adenylate cyclase system in the ovine fetal adrenal. PMID- 6279970 TI - The in vitro metabolism of estrone and estradiol-17 beta and their 3-sulfates by the renal tissues from pregnant and female fetal guinea-pigs. PMID- 6279971 TI - A mathematical model describing the subcellular localization of non-membrane bound steroid, seco-steroid and thyronine receptors. PMID- 6279972 TI - The role of chemotherapy as an adjuvant to surgery in the initial treatment of primary soft tissue sarcomas in adults. AB - A prospective study was carried out to determine the results of chemotherapy as an adjuvant to surgery in the initial treatment of adult primary soft tissue sarcomas. The results were compared with those in a group of patients with similar histologic types of primary sarcoma treated by surgery alone. The chemotherapy regimen consisted of adriamycin, 60 mg/m2 intravenously on Day 1, and DTIC, 250 mg/m2 on Days 1 through 5. The cycle was repeated every 22 days. The total dose of adriamycin was 500 mg/m2; the DTIC was continued for 1 year. The adjuvant chemotherapy group consisted of 113 patients (group one) and the concurrent surgical resection group consisted of 144 patients (group two). In group one, 53 tumors were T1 and 60 tumors were T2; 67 of the tumors were grade 3 and 46 were grade 4. In group two, 65 had T1 and 79 had T2 tumors; 82 tumors were grade 3 and 62 were grade 4. The anatomic location and histologic types were similar in both groups. Seventy-seven (77%) of the 113 patients in group one lived disease free for 2 years, compared with 59% of the 144 patients treated by resection alone. Of the patients eligible for 5-year survival analysis, 74% were disease free in the adjuvant chemotherapy group, compared to 50% in the surgical group. The incidence of local recurrence was about the same in both groups. From a histologic standpoint, malignant fibrous histiocytomas and myogenic sarcomas appeared to benefit most with use of the use of the two-drug regimen used here, and the least favorable response was in liposarcomas and fibrosarcomas. In other histologic types, the role of adjuvant chemotherapy needs further clarification. PMID- 6279973 TI - Oat cell carcinoma of the esophagus. AB - An oat cell carcinoma occurring in the esophagus of a 59-year-old man is described. A review of another 22 cases of such a tumor from the English literature is made. These tumors may have three histologic patterns; pure oat cell carcinoma, oat cell carcinoma with squamous cell carcinoma, and oat cell carcinoma with adenocarcinoma. They appear to arise from enterochromaffin cells; the neoplastic cells show argyrophilia and neurosecretory-type granules on electron microscopy. Resection has been the principal mode of therapy. Overall survival for these patients is about 6 months, with most patients dying of extensive metastatic disease. PMID- 6279974 TI - Increased pyrophosphate rib uptake: clinical correlation. AB - Two hundred and thirty-one patients had 270 bone scans. Increased rib uptake was correlated with radiological findings. Interpretation of results whether due to benign or to metastatic disease depends on other observations. These include whether the uptake is localized or along the rib outline, single or multiple, and associated with other abnormal uptake in the rest of the skeleton. Abnormal rib uptake was encountered in 23% of patients scanned for malignant disease and 10% of patients scanned for benign conditions. In the first group 34% were true positive and of these, rib lesions were multiple in 32% and solitary in 2%. In 20% the rib uptake was due to benign conditions and in 46% it was indefinite because of absent radiological findings. In this last difficult group 17% were due to probably early rib metastases and 29% were due to benign causes. Rib metastases were missed in only two cases, 1.1%. One case was multiple myeloma and the other was direct extension from underlying anaplastic lung carcinoma. PMID- 6279975 TI - Radiation therapy for superior vena caval syndrome due to lung cancer. PMID- 6279976 TI - Diet and urinary excretion of lignans in female subjects. AB - Lignans, a class of compounds having a 2,3-dibenzylbutane skeleton, have recently been identified for the first time in humans and animals and evidence indicating their formation by intestinal microflora has previously been established in rats and humans. In the present report the influence of diet on the biosynthesis of this new group of compounds was investigated by comparing the urinary excretion of the principal lignan, trans-2,3-bis-(3-hydroxybenzyl) -butyrolactone (enterolactone, HBBL), in 12 omnivoric and 14 vegetarian women. Young vegetarian women were found to excrete significantly greater amounts of enterolactone than omnivores, while old vegetarians excreted comparable amounts to the omnivore group. A statistically significant (P less than 0.01-0.001) correlation was found between the amount of fibre in the diet and the urinary enterolactone excretion. PMID- 6279977 TI - Factors modulating the activity of ornithine decarboxylase in rat HTC cells. AB - Dibutyryl cyclic AMP (dibu-cAMP), N1-acetylputrescine and both N1 acetylspermidine and N8-acetylspermidine regularly induce ornithine decarboxylase activity in both a time- and a concentration-dependent manner when added to HTC cells in suspension culture. N,N'-Diacetyl-1,6-diaminohexane and N-acetyl-1,3 diaminopropane, aliphatic analogues of acetylated putrescine containing the carboxamide bond but not known to occur naturally in animal tissues, are also effective inducing agents. The addition of 10(-2)M putrescine to the cell culture at the time of maximal induction of ODC in HTC cells by dibu-cAMP causes a marked decrease in the apparent half-life of ODC in less than 10 minutes. Ten-fold concentration of HTC cells at the time of their maximal induction by dibu-cAMP also results in an immediate decrease both in the specific activity of ODC and in its apparent half-life. When the population density of induced HTC cells is increased the relative effect of exposure of cells to increasing levels of putrescine is lessened. Some of the implications of these observations are discussed. PMID- 6279978 TI - Effect of polyamines on enzymes involved in DNA repair. AB - The influence of polyamines on various enzymes involved in the excision repair pathway of DNA, such as UV endonuclease, DNA polymerase I, DNA ligase and polynucleotide kinase, and two AP-endonucleases, were studied. The polymerizing activities of DNA polymerase I and polynucleotide kinase were found to be markedly affected by polyamines. In the former enzyme the effect can be attributed to the stabilization of the correct bihelical structure at the 3' end and in the latter case polyamines stabilize the polynucleotide kinase protein itself in the correct oligomeric structure. The effect of polyamines on the hydrolysis of apurinic and apyrimidinic sites in DNA and nucleosome particles were also investigated. Spermine and spermidine were found to be the most efficient polyamines in causing such hydrolysis both in the free DNA and in the nucleosome particles. PMID- 6279979 TI - Roles of polyamines in the replication of animal viruses. AB - Several animal viruses are known to contain significant amounts of polyamines but so far the function of these viral components is poorly understood. In this study the role of polyamines in the replication of two different types of viruses, herpes simplex virus type 2 and Semliki Forest virus (SFV) has been investigated. Purified SFV was found to contain fairly small amounts of polyamines, sufficient to neutralize only about 3% of viral nucleic acid phosphate, i.e., 1/20 of that found in herpes simplex virus. The production of both viruses was, however, markedly inhibited in BHK21 cells depleted of polyamines by treatment with alpha difluoromethylornithine, a specific inhibitor of polyamine synthesis. This inhibition was reversed by putrescine, spermidine and spermine, and at least partly reversed by several other diamines and polyamine homologs. The activity of viral RNA polymerase induced by SFV infection was markedly reduced in polyamine depleted cells but increased rapidly after addition of spermidine to the culture medium. It appears that the inhibition of virus production in polyamine-depleted cells is due in part to malfunction of protein synthetic machinery of the host cell. The possibility that other steps in virus synthesis and assembly are affected by polyamine deficiency is currently being investigated. PMID- 6279980 TI - [Insulin and glucagon receptors in the liver. Their regulation]. PMID- 6279981 TI - [The adsorption capability of beidellitic Montmorillonite]. PMID- 6279982 TI - [A comparative study of cephotaxime and other antimicrobial agents in relationship to Pseudomonas aeruginosa (author's transl)]. PMID- 6279983 TI - The LDL receptor defect in familial hypercholesterolemia. Implications for pathogenesis and therapy. AB - Familial hypercholesterolemia results from one of several genetic defects in a cell surface receptor that normally controls the degradation of low density lipoprotein. The clinical and genetic features and the pathophysiology of these defects are discussed. Knowledge of the regulation of LDL receptors in the liver can be exploited in the design of a physiologically rooted therapy for familial hypercholesterolemia. PMID- 6279984 TI - The dietary treatment of hyperlipidemia. Rationale, technique and efficacy. AB - Working from the foundation of the role of diet in the pathogenesis of hyperlipidemia, the authors present a rational, detailed therapy for treating the disorder. A three-phase approach is offered so that the patient can gradually incorporate new eating behavior into his or her lifestyle. PMID- 6279985 TI - Comparison of metronidazole assay by microbiological and chemical methods. AB - Chemical (thin layer chromatography/fluorescence quenching in situ) and microbiological (agar well, diffusion technique with Clostridium perfringens as indicator strain) methods of assaying metronidazole have been compared. On dummy samples made with pure metronidazole in pooled human serum, both methods had a coefficient of variation ranging from 5.5 to 9.6 per cent of the mean. The microbiological method slightly underestimated the real amounts, and also had lower values than the chemical procedure. Comparison of serum and urine samples taken during the early, middle and late periods after medication to volunteers showed that biotransformation to antibacterially active metabolites contributes significantly to the antibacterial activity, particularly in urine. Biotransformation explains why microbiologically determined concentrations were higher than those determined chemically in samples taken at least 16-20 hours after intake of tablets or suppositories. It is important to be aware of the circumstance that the results of microbiological assay are sensu strictu limited to the particular indicator strain used, since other bacteria may exhibit other patterns of sensitivity to metronidazole and its metabolites. PMID- 6279986 TI - The effect of temperature upon the responses of isolated cardiac muscle to sympathomimetic amines. PMID- 6279987 TI - [Bronchus cancer - surgery]. AB - Of 9,000 patients with bronchus carcinoma observed during 1958-1980, 3.041 (33%) underwent resection. Although in principle the standard operations are lobectomy for the peripheral and bilobectomy or pneumonectomy for the central carcinoma, parenchyma-saving operations such as clamping or segmental resection or main bronchus resection are justified under certain conditions; however, here all regional lymph nodes have to be removed again. We consider the surgical indication of small-celled carcinoma like any other form of bronchus carcinoma. For stage I, the 5-year survival chance is 40% - 50%, for all stages only 20%. Extended resections including pericardium, large vessels, thoracic wall, trachea, upper thoracic aperture, are justified especially in symptomatic cases, although the 5-year survival rate is less than 10% in these cases. PMID- 6279988 TI - [Chemotherapy of bronchogenic cancer (author's transl)]. AB - This report summarizes our experience with protocols "ACLI" and with alternative combinations such as ifosfamide plus etoposide in the chemotherapy of inoperable bronchogenic carcinoma. Using the "ACO"-protocols and consolidating irradiation, 77% complete remissions were achieved in patients with limited disease (median survival of 21 months). Patients with extensive disease (50% complete remissions) had a median survival of 10 months. The survival data point to a plateau after 30 months, suggesting that approximately 20% of limited disease patients may achieve long-term survival. Relapse patterns are analyzed. PMID- 6279989 TI - [Soft tissue tumors: pathology]. PMID- 6279990 TI - [Hyperthermic perfusion with cytostatic drugs -- an adjuvant therapy in soft tissue sarcoma of the limbs (author's transl)]. AB - Hyperthermic perfusion was used in 15 adult patients with soft tissue sarcoma of the limbs. Only patients were selected with prognostically unfavorable or recurrent tumors, which were excised with a limited safety margin. Three patients died from pulmonary metastases 16, 24 and 34 months after perfusion. In 2 patients a local recurrence developed. Ten patients are alive and free of recurrence. The follow-up period ranges from 7 - 47 months with a median of 30 months. These results encourage the further use of hyperthermic perfusion. PMID- 6279991 TI - [Medical documentation on surgery and follow-up (author's transl)]. PMID- 6279993 TI - [Bronchial carcinoma, bronchial microcarcinoidosis (tumorlets), and bronchial carcinoids (author's transl)]. AB - During the last 22 years in Styria lung cancers, as revealed in autopsies, have increased in men by 9% and in women by 173%. In women undifferentiated carcinomas have been found six times as frequently as previously. The histological findings of mixed types of carcinomas with confusing cytology are presented: metastasizing squamous carcinoma with a survival time of 14 years and lung cancer developing in the upper lobe 35 years after being present in the lower lobe. In oat-cell carcinomas, certain structures suggest they originate from endocrine cells of the bronchial epithelium. This was also proven in bronchial peptide-producing microcarcinoidosis (tumorlets) and in carcinoids. At the same time, there are neuroendocrine contacts between tumors and nerves similar to carcinoids of the appendix and rectum. PMID- 6279992 TI - [Second generation of absorbable suture material. Tissue reaction and indication (author's transl)]. AB - Polyglycolic acid (PGA) sutures have proved to be superior than other, nonabsorbable suture materials: a higher traction capacity and knot-tearing security, a defined and nearly linear decrease of traction capacity, no immunological reactions, very little capillarity, minimal tissue reaction, nearly no scar formation, and a bactericidal effect. PGA sutures of the second generation have the same advantages, but surgical handling is so much improved that they may be technically employed in unfavorable locations. In the surgery of gastrointestinal tract and respiratory organs there is a trend to connect all layers with one suture. PMID- 6279994 TI - [Bronchial cancer - cytology]. PMID- 6279995 TI - Chemotherapy for salivary gland cancer. AB - Salivary gland cancers are usually treated with surgery and irradiation; however, some of the aggressive salivary gland cancers recur or metastasize and are not amendable to treatment with further surgery or irradiation. Little is known about chemotherapy for these palliative situations because of the relatively scarcity of these cancers. The data in the literature has provided little clinical information because all salivary gland cancers are usually lumped together and/or multiple different drug combination are used. In an effort to arrive at a rational basis for recommending specific drug regimens for specific histologic types of salivary gland cancers, a two-part study was undertaken to determine which chemotherapy drugs seem to be effective or ineffective. One part was a literature review, and the second part was a survey of numerous institutions' experiences, including our own. A total of 85 cases of salivary gland cancers treated with chemotherapy were felt to be evaluable for this study. The overall response rate (complete and partial) was 42%. Although disease responded whether it was local, regional, or distant disease, there was a higher response rate in local-regional disease compared to distant metastases. Salivary gland cancers are definitely sensitive to chemotherapy drugs. This study reveals which drugs seem to be effective and provides some rational basis for future chemotherapy trials for salivary gland cancers. PMID- 6279996 TI - Cerebral endothelial cell culture. I. The presence of beta 2 and alpha 2 adrenergic receptors linked to adenylate cyclase activity. AB - Cultured endothelial cells derived from cerebral microvessels separated from 2 day-old rat brain contain a specific beta 2 and alpha 2-adrenergic sensitive adenylate cyclase (AC). Among the various tested hormones, PGE1 and PGE2 were found to be the most potent activators, while adenosine, angiotensin I and II, gamma-aminobutyric acid and vasoactive intestinal peptide inhibited the enzyme activity. However, acetylcholine, histamine, serotonin, glycine, glutamine, bradykinin, neurotensin and vasopressin (Lysine and Arginine) had no effect on the adenylate cyclase activity in this model. The susceptibility of the cerebrovascular endothelial AC system to the vasoactive substances as well as presence of beta 2 and alpha 2-type adrenergic receptors in the cultured endothelium provides additional support for the proposed endothelial involvement in the regulation of cerebrovascular permeability and blood flow. PMID- 6279997 TI - Evidence that systemically administered salbutamol reduces food intake in rats by acting on central beta-adrenergic sites. AB - Salbutamol was found to reduce food intake in a dose-related manner. The effect was prevented by the beta-adrenolytic drugs d,1-propranolol and d,1-alprenolol. In contrast, phentolamine, penfluridol and metergoline, which block alpha adrenergic, dopamine and serotonin receptors respectively, or bilateral lesions of the ventral noradrenergic bundles were not able to counteract salbutamol's effect. The reduction of food intake induced by salbutamol was prevented by intracerebroventricular administration of d,1-propranolol. In addition, salbutamol was found to effectively reduce the apparent motivation for food as revealed by its effect on food-rewarded runaway behaviour. This effect was also counteracted by d,1-propranolol pretreatment. The findings indicate that food intake may be reduced by peripheral administration of salbutamol and that the effect is mediated by stimulation of central beta-adrenergic sites. PMID- 6279998 TI - Treatment with antidepressants and histamine receptor mediated 3H-cyclic AMP formation in guinea pig cortex. AB - It has been recently reported that most of the antidepressant drugs block histamine H1 and H2 receptors in the brain under in vitro conditions and it has been suggested that this may be related in part to their therapeutic effect. Since the in vitro and in vivo effects of these drugs may differ, we studied the effect of treatment with antidepressant drugs on histamine receptor sensitivity in the guinea pig brain and observed that chronic treatment with tricyclic antidepressants or phenelzine (an MAO inhibitor) causes a reduction in histamine receptor sensitivity. This reduction is probably mediated through two different mechanisms, since only tricyclic antidepressants cause a reduction after acute treatment. Although some of the side effects of antidepressant treatment may be related to the blockade of histamine receptors, these results do not support the assumption that this effect of antidepressant treatment contributes to their clinical effects. PMID- 6279999 TI - Stimulatory and protective effects of benzodiazepines on GABA receptors labeled with [3H]muscimol. AB - We investigated the effects of benzodiazepines on [3H]muscimol binding to rat brain membranes and on heat inactivation of GABA receptors. Scatchard analysis of [3H]muscimol binding to frozen and 0.05% Triton X-100 treated membranes revealed two components; a higher affinity (Kd = 2.2 nM, Bmax = 1.2 pmol/mg protein) and a lower affinity component (Kd = 15.9 nM, Bmax = 4.4 pmol/mg protein). Diazepam and flurazepam (3 microM) increases significantly the specific binding of 40 nM but not of 2 nM [3H]muscimol. This stimulation was attributed to an increase in the affinity of the lower affinity component for GABA receptors. The time course of heat inactivation of GABA receptors revealed rapidly and then slowly denaturating Phases. These observations would suggest that there are multiple GABA receptors with different sensitivities to the heat treatment. Diazepam depressed remarkably the slowly denaturating phase(s). After heat treatment for 50 min, the single component of GABA receptors with Kd of 14.3 nM and Bmax of 0.6 pmol/mg protein survived, whereas in the membranes preincubated with 3 microM diazepam, the Kd and Bmax of the still viable GABA receptors were 14.8 nM and 1.14 pmol/mg protein, respectively. In light of these findings, the stimulation of the lower affinity component of GABA receptors may be related to the protective effects of these drugs against heat inactivation. PMID- 6280000 TI - Development of a radioiodinated ligand for characterising alpha 1-adrenoceptors. AB - Two alpha-adrenoceptor antagonists, phentolamine and 2-(beta-(4-hydroxyphenyl) ethylaminomethyl)-tetralone (BE 2254) which are phenolic derivatives were radioiodinated after chloramine-T oxidation of Na125I and the labelled material isolated by chromatography. 125I-phentolamine does not bind selectively to alpha adrenoceptors in guinea pig brain whereas the 125I-BE 2254 derivative binds rapidly, reversibly and with high affinity to these receptors with a Kd of 230 pM. At low concentrations of 125I-BE 2254 (less than 100 pM) approx. 90% of the bound radioligand is specifically bound and under these conditions drug displacement studies show that the ligand binds predominantly to the alpha 1 subclass of adrenoceptors. Binding measurements to kidney and smooth muscle membrane preparations indicate that 125I-BE 2254 may also be a useful tool in the study of alpha-adrenoceptors in peripheral tissues. The high specific activity of 125I-BE 2254 permits the use of minimal quantities of membrane material for receptor assay and ligand displacement measurements, e.g. 250 micrograms per assay tube, and this provides a significant advantage over the use of existing radioligands such as 3H-prazosin which requires approx. 40 times as much tissue. PMID- 6280001 TI - Alpha and beta adrenergic receptors of canine lung tissue identification and characterization of alpha adrenergic receptors by two different ligands. AB - Adrenergic receptors of canine peripheral lung tissues were measured by direct binding techniques using [3H]dihydroergocryptine ([3H]DHE), [3H]prazosin and [3H]dihydroalprenolol ([3H]DHA). All three ligands bound to canine lung tissue with saturability, stereospecificity and reversibility. Adrenergic agonists competed for binding of [3H]DHE and [3H]prazosin in the order: 1-epinephrine greater than 1-norepinephrine greater than d-epinephrine greater than d norepinephrine greater than 1-isoproterenol. Adrenergic antagonists competed for binding of [3H]prazosin in the order: prazosin greater than phentolamine greater than yohimbine. Inhibition curves of [3H]DHE by prazosin or yohimbine were biphasic suggesting two subtypes of binding sites. Maximum binding capacities of [3H]DHE ranged from 30.6 to 42.7 fmol/mg protein. [3H]prazosin from 18.3 to 26.9 fmol/mg protein and [3H]DHA from 135.2 to 359.4 fmol/mg protein. When both [3H]DHE and [3H]prazosin were used in the same membrane preparation, specific binding of [3H]DHE was always more than that of [3H]prazosin. Since [3H]prazosin is considered to bind to alpha 1 adrenergic receptors specifically and [3H]DHE is considered to bind alpha 2 adrenergic receptors nonselectively, the difference between the numbers of the specific binding sites of these two ligands should represent alpha 2 adrenergic receptors. Alpha 2 adrenergic receptor density ranged from 9.5 to 21.1 fmol/mg protein. Our results suggest the existence of both alpha 1 and alpha 2 adrenergic receptors in canine peripheral lung tissue. Approximately 40% of alpha adrenergic receptors were alpha 2. The ratio of alpha/beta adrenergic receptors ranged from 1:3.3 to 1:10.4. The ratio of alpha 1/beta adrenergic receptors ranged from 1:6.7 to 1:21.1. PMID- 6280002 TI - High performance liquid chromatography of pharmacologically active amines and peptides in biological materials. AB - Precise and quantitative reversed-phase high performance liquid chromatographic (HPLC) procedures are described which can be used in biogenic amine and neuropeptide research. The amine procedure was applied to various pharmacological matrices including plasma, heart tissue and brain. The use of peptide HPLC as an analytical tool for various neuropeptides is illustrated by studies on des tyrosine-gamma-endorphin (DT gamma E) metabolism in the brain and the stability of an ACTH (ORG-2766) analogue during a chronic infusion in rats. The power of HPLC as a research tool in peptide pharmacology is described, discussed and demonstrated as an aid in the understanding of the pharmacological effects of exogenous peptides and the function of the brain. PMID- 6280003 TI - Clonidine withdrawal: activation of brain noradrenergic neurons with specifically reduced alpha 2-receptor sensitivity. PMID- 6280004 TI - Effects of chronic reserpine pretreatment on myocardial sodium pump and inotropic sensitivity to ouabain in guinea pig heart. PMID- 6280005 TI - Aging and benzodiazepine binding in the rat cerebral cortex. AB - The specific binding of 3H-diazepam in the cerebral cortex was investigated in membrane preparations from 6 and 30 month old Fisher-344 rats. No age-related differences in the association, equilibrium, or dissociation, binding characteristics were observed. The increased sensitivity of the elderly to the central sedative effects of the benzodiazepines does not, therefore, appear to involve changes in binding to the receptor site located in the cortex. PMID- 6280006 TI - Platelet-derived growth factor increases prostaglandin production and decreases epidermal growth factor receptors in human osteosarcoma cells. AB - Human platelet-derived growth factor (PDGF) stimulated the production of prostaglandin E2 (PGE2) by G-292 cells, a clonal line of human osteosarcoma cells. Half-maximal stimulation occurred with 9 ng/ml PDGF and maximal stimulation, 3-fold above control values, occurred with 40 ng/ml of the protein. Treatment of G-292 cells with 40 ng/ml PDGF also reduced the binding of iodinated epidermal growth factor (EGF) to the EGF receptor on G-292 cells. The effect was time-dependent, and EGF binding was reduced to 60% of control by 24-48 h. PDGF did not, however, compete directly for binding to the EGF receptor. The effects of PDGF and EGF on increased PGE2 production appeared to be additive at all concentrations tested, indicating that they may act through a common pathway, but not via the same membrane receptors. PMID- 6280007 TI - CGS 8216: receptor binding characteristics of a potent benzodiazepine antagonist. AB - CGS 8216 is a novel nonbenzodiazepine that inhibited 3H-flunitrazepam (3H-FLU) binding to rat synaptosomal membranes in vitro at subnanomolar concentrations. It prevented the in vivo labeling of brain benzodiazepine receptors by 3H-FLU with the same potency as diazepam when given orally to mice. Pharmacologic tests showed that it was devoid of benzodiazepine-like activity but it antagonized the actions of diazepam in these tests. It did not interact with alpha- or beta- adrenergic, H1-histaminergic or GABA receptors but it inhibited adenosine activation of cyclic AMP formation. Studies with 3H-CGS 8216 demonstrated that it bound specifically and with high affinity to rat forebrain membranes and was displaced by drugs with an order of potencies similar to that observed when 3H diazepam and 3H-FLU were used as radioligands. The regional distribution of 3H CGS 8216 binding sites in the brain was also similar to that of 3H-FLU. Dissociation of 3H-CGS 8216 binding was slow at 0 degrees C but increased with temperature and was almost complete within 1 min at 37 degrees C. Scatchard analyses were linear, yielding KD values of 0.044, 0.11 and 0.18 nM at 0, 25 and 37 degrees C, respectively; the Bmax value did not change appreciably with temperature and was approximately 1000 fmoles/mg protein. Using 3H-FLU, the value for Bmax as well as for the KD increased with temperature. The total number of binding sites determined for 3H-FLU was greater than that for 3H-CGS 8216 at each temperature. CGS 8216 exhibited mixed-type inhibition of 3H-FLU binding. GABA did not stimulate 3H-CGS 8216 binding whereas it enhanced 3H-FLU binding. CGS 8216 may be a useful ligand for probing the antagonist properties of the benzodiazepine receptor and is likely to exhibit interesting therapeutic effects. PMID- 6280008 TI - Variations in the number of pituitary LHRH receptors correlated with altered responsiveness. AB - Isolated pituitary cells from metestrous, ovariectomized (OVX), and ovariectomized-estradiol treated (OVX-EB) rats were employed to study the gonadotrophin response to luteinizing hormone-releasing hormone (LHRH) challenge and to quantitate LHRH receptors, using a labeled LHRH analog. Ovariectomy (3-4 weeks post castration) resulted in a reduction of LHRH receptor concentration from 34.4 +/- 2.1 in metestrous females to 14.3 +/- 0.9 fmoles/10(6) cells. Concomitantly, the luteinizing hormone (LH) response to a near-maximal dose of LHRH (5 ng/ml) decreased from a 3-fold stimulation in intact females to 1.13-fold stimulation in cells from OVX rats. Replacement therapy with EB (50 ug/rat for 2 days) to OVX rats restored LH response and LHRH binding sites (a 2.5-fold stimulation in LH secretion and 32.0 +/- 2.1 fmoles/10(6) cells, respectively). The LH response to LHRH stimulation was not altered after one day of EB treatment although the number of LHRH binding sites was increased. The changes in the number of LHRH binding sites were not accompanied by any alterations in the affinity of the LHRH analog (K3 approximately equal to 0.5 X 10(-9)M). It is concluded that variations in LHRH receptor number reflect the degree of pituitary sensitivity to LHRH and it may suggest that LHRH and estradiol modulation of gonadotropin release is mediated by these receptors. PMID- 6280009 TI - Role of a humoral sodium-potassium pump inhibitor in experimental low renin hypertension. AB - Recent evidence suggests that the vascular sodium-potassium pump suppression previously observed in animals with various models of low renin hypertension results from a circulating heat stable ouabain-like agent. It appears to come from or be influenced by the anteroventral third ventricle area of the brain and its action on blood vessels results in depolarization of the smooth muscle cell. Suppression of the vascular sodium-potassium pump, with ouabain for example, increases contractile activity and the contractile responses to vasoactive agents. Thus the humoral pump inhibitor may be involved in the genesis and maintenance of experimental low renin hypertension. PMID- 6280010 TI - Impairment of renal compensatory hypertrophy by hypothyroidism in the rat. AB - Renal compensatory hypertrophy (RCH) occurs in hypothyroid rate, but it is impaired when compared to RCH found in euthyroid controls. It is due to cellular hypertrophy as the DNA content does not change and the Protein/DNA ratio increases in the compensating kidney. RCH is enhanced by thyroxine (T4) with a rise in the DNA content of the compensating kidney, but the Protein/DNA ratio does not change indicating that hypertrophy is as important as hyperplasia. Corticotrophin (ACTH) given to eu and hypothyroid rats enhances RCH with an increase in the protein content of the compensating kidney without any change in its DNA content. In the hyperthyroid rats, the enhanced RCH is not further increased by ACTH and the rise in the kidney DNA content elicited by T4 is suppressed by ACTH. The Protein/DNA ratio is increased by ACTH in hypo, eu and hyperthyroid rats. The renotrophic action of ACTH is due to hyperadrenocorticism: it is related to an increased plasma testosterone level and to a disturbed Na+, K+ and glucose metabolism. PMID- 6280011 TI - Mechanisms altered beta-adrenergic responsiveness in the hyperthyroid and hypothyroid turkey erythrocyte. AB - Studies on the relationship between thyroid hormone and the beta-adrenergic catecholamines have been carried out in the turkey erythrocyte. Conditions of thyroid hormone excess and deficiency were examined with respect to their effects on the beta receptor itself, as well as to their effects on associated biochemical and physiological indices of beta receptor function, including agonist stimulated adenylate cyclase activity, cellular cyclic AMP generation, and catecholamine-induced stimulation of potassium ion influx. Erythrocytes obtained from hypothyroid turkeys showed a marked (approximately 50%) reduction in beta receptor number without any change in receptor affinity for agonists or antagonists. Catecholamine-sensitive adenylate cyclase activity and cellular cyclic AMP levels were similarly reduced. The sensitivity of these cells to agonist-stimulated potassium influx was significantly decreased, but maximal agonist-stimulated transport rate was unchanged. Analysis of the quantitative relationship between beta receptor number, agonist concentration, and level of catecholamine-stimulated potassium influx indicates that, at any given absolute level of receptor occupancy, the level of agonist-stimulated potassium influx is identical in hypothyroid and normal erythrocytes, and that the diminished physiological sensitivity of the hypothyroid cell is attributable in its entirety to a reduction in beta receptor number per se. The results obtained in the hyperthyroid turkey erythrocyte were strikingly different. Here, beta receptor number, binding affinity for agonists and antagonists, catecholamine-sensitive adenylate cyclase activity, and maximal cyclic AMP levels were all unchanged. In contrast, maximal agonist-stimulated potassium ion transport was markedly reduced, while the concentration of isoproterenol required for half-maximal stimulation was only slightly increased. Analysis of the relationship between beta receptor number, agonist concentration, and catecholamine-stimulated potassium influx rate indicates that, at all absolute levels of beta receptor occupancy, the stimulation of monovalent cation influx is markedly blunted in the hyperthyroid cell. In contrast to the findings in the hypothyroid cell, where decreased physiologic sensitivity to catecholamines is directly attributable to a reduction in beta receptor number, the primary abnormality responsible for diminished catecholamine responsiveness in the hyperthyroid cell would appear to be located at a point "distal" to the beta receptor itself. PMID- 6280012 TI - Studies on the mechanism of thyroid hormone stimulation in vitro of human red cell Ca2+-ATPase activity. AB - The stimulation in vitro of human red blood cell Ca2+-ATPase activity by thyroxine (T4) and triiodothyronine (T3) in physiological concentrations is shown to depend upon binding of iodothyronines to red cell membranes. Calmodulin enhances the activity of thyroid hormone in this model system but there is no direct interaction of calmodulin and hormone. PMID- 6280013 TI - Isolation and diagnostic cultivation of Entamoeba histolytica: a comparative study of two methods. PMID- 6280014 TI - A role for amines in the antidepressant effect of exercise: a review. AB - A review of the literature suggests that exercise may have antidepressant effects and, like other treatments for depression such as electroconvulsive therapy (ECT), antidepressant medication, and REM sleep deprivation, may enhance aminergic synaptic transmission in the central nervous system. In addition, the effects of exercise and other antidepressants on sleep are similar. Therefore, it is suggested that exercise is an antidepressant that enhances aminergic synaptic transmission in the central nervous system. Many more psychological and physiological studies must be performed in order to verify and quantify this relationship. Present statements that single out norepinephrine, dopamine, or serotonin as the crucial amine may be premature and oversimplified. Future physiological studies must take into consideration the advantages and disadvantages of human and animal subjects. Future psychological studies should be attentive to possible differences in psychological benefits between normal and depressed subjects and should not neglect the possible role of cognitive factors such as subjects' attitudes towards exercising or the feelings of accomplishment that may result from increased physical fitness. There is also a need to measure antidepressant effects in long-term exercise programs and in studies employing various forms of exercise. PMID- 6280015 TI - [A disseminated form of Histoplasma duboisii histoplasmosis in a Zaire patient (author's transl)]. AB - Report of a case of histoplasmosis observed in Zaire with lymphatic, pulmonary and intestinal localizations. Histoplasma duboisii was abundant in the stools, sputum and lymphatic tissue. With no amphotericine B available, treatment first started with high dosage of miconazole which had no effect. Ketoconazole have been, then, tried, giving a clinical and biological improvement, but patient still in a very bad condition, died. Ketoconazole seems to have a rapid but not lasting effect, even with increased dosages. PMID- 6280016 TI - [Viral hepatitis. Modern aspects of serological diagnosis and prevention]. PMID- 6280017 TI - Comparative study of Junin and herpes simplex viruses in mouse brain monolayer cultures. PMID- 6280018 TI - [Immunization of guinea pigs against Argentinian hemorrhagic fever with Tacaribe virus replicated in human diploid cells]. PMID- 6280019 TI - [Effects of ionizing radiation on the content of total collagen and its fractions and the activity of collagenolytic enzymes in rat tissues]. AB - Changes in collagen fractions as well as in the activity of leucocytic collagenase and collagenolytic cathepsin of rats after 30 days of gamma-rays exposure were examined. A statistically significant decrease in total collagen content resulting from the reduction of salt--soluble (NSC) and acid--soluble (ASC) collagen fractions was found. An increased content of insoluble collagen fraction (ISC) may confirm the opinion about stimulative gamma-rays influence upon cross--links formation. An increase in collagenolytic enzymes activity may account for an elevated degradation of collagen; with a contribution of leucocytic collagenase and collagenolytic cathepsin. Perhaps the quantitative increase in insoluble collagen stimulates elevated biosynthesis of collagenolytic enzymes determining the process of protein degradation. Further investigations are needed to explain the radiobiological effects of gamma-rays on the connective tissue. PMID- 6280020 TI - [Diagnostic value of antabuse and CBJ-131 test in workers chronically exposed to carbon disulfide]. AB - The studies covered 32 workers chronically exposed to CS2 and controls. In all subjects diethyldithiocarbamaturia was evaluated and J131 labelled Bengal Rouge decay half-time was determined. The studies indicated statistically significantly lower DDC excretion in CS2 exposed workers, as compared to controls. On the other hand no statistically significant differences were found at the Bengal rouge decay half--time in blood. The above studies suggest that the disulphuran metabolic route was impaired prior to the occurrence of the changes in liver blood flow of CS2 exposed workers. PMID- 6280021 TI - [Effect of passage on the fatty acid composition of the lipids in Clostridium perfringens A]. PMID- 6280022 TI - [Antiviral activity of Kalanchoe sap]. PMID- 6280023 TI - [Selection and mutant properties of Pichia guilliermondii yeasts with derepressed GTP cyclohydrolase, the enzyme of the 1st step in flavinogenesis]. AB - A positive method is proposed for selecting Pichia guilliermondii mutants with derepressed GTP cyclohydrolase. Mutants with the incompletely blocked gene RIB2 were used as parent strains; these can grow in a medium without riboflavin (RF) only if the enzyme is derepressed as the result of iron deficiency in cells. Strains growing in a medium without RF at the optimal supply of cells with iron were selected as regulatory mutants. The mutants accumulated 6,7-dimethylpterin in high concentrations and a small amount of RF in the medium and in the cells. The activity of GTP cyclohydrolase rather than that of RF synthase increased in the mutants; the activity of RF kinase and FAD pyrophosphorylase was not elevated. Hybrids produced by crossing the regulatory mutants with wild type strains did not accumulate 6,7-dimethylpterin in the medium and the activity of the GTP cyclohydrolase did not increase; this is indicative of the negative regulation for the expression of the structural gene for GTP cyclohydrolase. The authors propose a model for the regulation of GTP cyclohydrolase and RF synthase at the gene level involving iron ions as a corepressor. PMID- 6280024 TI - Federal support of radiological research. AB - Pervading the plans and objective outlined herein for continued and enhanced federal support of research in radiology is a challenge of unparalleled magnitude, for the economic foundation on which this support is based has rarely been more precarious. The new administration in Washington may well be the most fiscally constrained in half a century, and its stated interest in reducing federal expenditures could have disastrous consequences for the scientific research effort in this country, including that in radiology and the radiological sciences. The circumvention of these consequences may well require the dedicated effort of the entire scientific community over the next few months and years, including that part representing radiology and the radiological sciences. PMID- 6280025 TI - Good vibrations: Abrams's oscilloclast and the instrumental cure. PMID- 6280026 TI - [Wandering genes: fatal antibiotic-resistance]. PMID- 6280027 TI - [ACTH in the dexamethasone suppression test]. AB - Despite the great clinical importance of the low-dose dexamethasone suppression test (determination of cortisol after dexamethasone 2 mg p.o. at 11.00 p.m. in the evening before) and of the determination of ACTH in plasma, so far only little is known about the ACTH response to dexamethasone in normal subjects. In the present study cortisol and ACTH were determined by radioimmunoassay in normal subjects at 8.00 a.m., 10.00 a.m. and noon before and after dexamethasone. Additionally the cortisol response to ACTH was tested under both conditions. The ACTH concentrations in the morning show a periodicity quite similar to the well known circadian rhythm of cortisol secretion. Dexamethasone blocks this periodicity without decreasing ACTH significantly. The cortisol response to ACTH after dexamethasone is normal. In the morning only a basal and presumably biologically inactive ACTH is secreted which can not further be suppressed by dexamethasone. Therefore it is advisable for the diagnosis of hypercortisolism to measure ACTH only in stimulation tests, e.g. insulin-induced hypoglycemia. PMID- 6280028 TI - [Research in the treatment of fungal infections]. PMID- 6280029 TI - Meningeal carcinomatosis in small cell carcinoma of the lung. AB - Small cell carcinoma of the lung is extremely sensitive to chemotherapy and radiation therapy. We describe the case of a patient who had a complete response to such treatment but relapsed with meningeal carcinomatosis. We propose that prophylactic therapy to the spinal cord as well as the brain should be considered in the treatment of patients with small cell carcinoma of the lung. PMID- 6280030 TI - [Structure of human globin genes]. AB - An attempt has been undertaken to review the present data on the structure of alpha- and beta-like human globin gene families, their quantity in a cell genome, intergene spacers, evolution and common features and differences in globin genes sequences. Some other questions, concerning the regulation of globin genes expression, transcription, structure and processing of primary transcripts, the structure of globin messenger RNAs have been also discussed. PMID- 6280031 TI - [Electron transfer in hemoproteins. VIII. Influence of ionic strength on the rate of reduction of ferricytochrome c by oxymyoglobin derivatives, chemically modified at histidine residues]. AB - The influence of chemical modification of His residues in Mb on the rate of redox reaction in system MbO2--Cyt c has been studied at different ionic strengths and pH medium. The products of alkylation of all available His by bromacetate and iodacetamide, CM-Mb and CA-Mb, respectively, and myoglobin, modified by spin label 2,2', 6,6'-tetramethyl-4-bromoacetoxypiperidine-1-oxyl (SL) at His residue A10--Sl (His-A10)--Mb have been studied. It has been shown, that the character of the ionic strength dependence of reaction SL(His-A10)--MbO2 with Cyt c at pH 6.0 ann 7.0 is basically analogues to that, observed for intact protein. It means that only His-GH1 of two His residues, His-A10 and His-GH1, situated in the region of "active contact" of Mg with Cyt c molecule, participates in the interactions, essential for electron transfer. The interaction of the charge of this His with the negatively charged group of Cyt c is necessary, probably for the proper arrangement of other interactions in the active complex, because the deprotonation of His-GHl in the studied pH interval decreases the rate of the process by more than one order of magnitude. The rate of oxidation of MC-MbO2 and CA-MbO2 by ferricytochrome c, in contrast to intact protein, shows a weak dependence on the ionic strength and does not depend on the pH medium, throughout the range of ionic strengths from 0.005 to 1.0. The cause of the radical change in the ionic strength dependence is, probably, nearly entire disturbance of electrostatic interactions in the active complex due to chemical modification of His residues in the site of "active contact", and first of all, the His-CHl residue. The fact, that during alkylation of all available His in Mb the electron transfer persists in the system, points to that in the process of electron transfer to cytochrome c, uncharged group, most probably "inner" His-B5, participates. Based on the data on spatial structure and the obtained results, the positions of the charged groups in the site of "active contact" of Mb with Cyt c molecule are presented. PMID- 6280032 TI - [Molecular mechanisms of stabilization and preparation of metabolic breaks in DNA strands in vivo. I. Tandem action of exonuclease V and DNA polymerase II]. AB - Investigation of the formation of metabolic imbalance breaks in the DNA of thy- cells of E. coli during thymine starvation is described. The results of experiments indicate that two enzymes--exonuclease V and 3' exonuclease activity of DNA polymerase II take part in the formation of metabolic imbalance gaps. The manifestation of activity of the enzymes has a tandem character. PMID- 6280034 TI - [ESR-investigation of pea mitochondria iron-sulfur centers]. PMID- 6280033 TI - [Molecular biology criteria for functioning of the genome of Rous sarcoma virus in transformed rat XC cells]. AB - Integration, transcription and translation of Rous sarcoma virus (RSV) genome was analyzed in transformed rat fibrosarcoma XC cells (non-virus producing). About ten copies of RSV genome were integrated into the host cell genome. Different types of viral RNA were identified in the cytoplasmic fraction whose sedimentation values were nearly the same as for RSV-specific RNA in virus producing chicken cells. These RNA were translated in polysomes and directed synthesis of gag, env and src proteins. In comparison to chicken cells in transformed XC cells the cleavage of gag precursor Pr 76gag could not be observed. PMID- 6280036 TI - [Simian adenovirus genome. II. Identification of specific fragments of viral DNA possessing transforming activity]. AB - The antigenic variant of simian adenovirus 7 (SA7) DNA was cleaved by restriction endonucleases EcoRI, XbaI, BamHI, SalI. The resulted digests of viral DNA were tested for transforming activity using the "calcium" technique. It was shown that BamHI. XbaI and SalI digests transformed primary baby rat kidney cells as well as native viral DNA. The transforming activity of separated BamHI and SalI fragments was tested also. The viral DNA fragments with transforming activity (BamHI-B and SalI-B) were localised on the left of the physical map of the viral genome. It was also shown that fragment-transformed cell lines were able to form colonies in 0.33% agarose medium. PMID- 6280035 TI - [Cloning and study of inserted sequences of gene 28S in Drosophila melanogaster ribosomal RNA]. AB - Cloning of fragments of ribosomal genes containing insertions in the 28S RNA gene has been reported earlier. Subcloning of DNA fragments corresponding to insertion sequences and their hybridization with DNA, RNA and polytene chromosomes from different flies is described. Type 1 insertions (containing BamI sites) are highly heterogeneous in length and sequence even in homozygotes. Type 2 insertions (with EcoRI sites) are rather homogeneous. Two types of insertions are represented in the D. melanogaster genome by 50 and 30 copies, respectively. Restriction fragments with insertions significantly differ in DNA from embryos and larvae. D. simulans and D. virilis also contain the sequences of both types of insertions, though in fewer number of copies. Type 1 insertions seem to be poorly transcribed, and type 2 insertions are not transcribed at all. Among 2000 recombinant clones screened a number of DI plasmids hybridizing to isolated insertions were obtained. Six of them were mapped with restriction endonucleases and hybridized with insertion fragments. rRNA and polytene chromosomes. All of these DI plasmids hybridize with the nucleoli, one with the chromocenter and one with the 79F 3L site. In LI9, not coding for rRNA, the sequences, corresponding to two types on insertions are located only a few kilobases apart. D17a does not encode for rRNA, but hybridizes in situ only with the nucleoli. PMID- 6280037 TI - [Study of the irreversible conformation change of immunoglobulin M by spin labeling at the carbohydrate and peptide moieties of the molecule]. AB - The irreversible conformational change of the immunoglobulin M (IgM) molecule (Waldenstrom disease) at pH approximately 3 was studied by means of spin-labels introduced in the carbohydrate (2,2,6,6,-tetramethyl-4-aminopiperidine-1-oxyl) and peptide (2,2,5,5,-tetramethyl-3-(dichloro-symm.-triazinylamino)-pyrrolidine-1 oxyl) moieties of the molecule. A marked rise of structure density of IgM especially in the (Fc)5-region and some minor local conformational changes in the Fab-regions were found. Comparison of our findings with the published data shows that Fab-regions of the principal immunoglobulins are rigid structures. Steric hindrance for Fab-regions increases markedly in the row Fab--F(ab')2--IgG--IgA- IgM restricting their spatial mobility. Monomeric Fc-regions of IgM are evidently flexible and one of the domains is especially mobile. It is supposed that oligosaccharide groups of IgM are of two types which differ in their spatial mobility. It was found by ammonium sulfate precipitation of IgM spin-labeled at the peptide moiety that the relative mobility of amino acid residues coupled with spin-label is strongly restricted. PMID- 6280038 TI - Cytomegalovirus encephalitis in children. PMID- 6280040 TI - The cytomegalic inclusion cell and disease. II. Social behaviour in vitro. AB - Upon primary cytomegalovirus infection, previously healthy individuals acutely reject the mass of virus-infected cells, by adequate cellular and humoral defence mechanisms. By contrast, the immunologically immature or compromised host, while still able to neutralize extracellular virus, will tolerate the persistence of the allenated population of cytomegalovirus-converted cells. The micropathology of cytomegalovirus-infected monolayers, however, runs a different course, due to the absence of any immune effector in the in vitro systems. In cultured monolayers of fibroblasts, the infected cells are selected out and aggregate engulfing one another to form giant multinucleated structures. PMID- 6280039 TI - Polymorphism of nuclear inclusions in carcinoma of the cervix uteri. AB - Frequency, polymorphism and ultrastructural characteristics of the nuclear inclusion bodies encountered in cancers of the uterine cervix are reported and briefly discussed. The nuclear inclusions are grouped in three distinct types: a) nuclear bodies (comprising type I and II inclusions according to Bouteille et al.'s classification), b) inclusion of cytoplasmic origin, and c) particles of chromatic material. The ultrastructural aspects of the chromatic particles suggest an early structuration of viral chromatin into core material. There appears to be a direct relation between the frequency of chromatic particles and raised antiherpetic antibodies in the patient's sera. PMID- 6280041 TI - Electrical stimulation of denervated muscle prevents decreases in oxidative enzymes. AB - The influence of muscular contraction on the oxidative enzymes and the diameters of muscle fibers was investigated. Soleus muscles of guinea pigs were denervated for four weeks. The denervated fibers showed a reduction in the intensity of staining for beta-hydroxybutyrate dehydrogenase, cytochrome oxidase, succinate dehydrogenase, and NADH-dependent tetrazolium reductase. Denervation also resulted in a decrease in fiber diameter. Denervated soleus muscles were electrically stimulated to contract over a four-week period at a frequency normally received by slow contracting muscles. Electrical stimulation caused the stain intensity of histochemical reactions for oxidative enzymes to appear to be normal or greater than normal in 90% of the denervated fibers. Stimulation also caused 69% of the denervated fibers to be of normal or greater than normal size. The results demonstrate that contraction of denervated muscle by electrical stimulation prevents the loss of oxidative enzymes and the atrophy associated with denervation. PMID- 6280043 TI - Chronic elevation of endoneurial fluid pressure is associated with low-grade fiber pathology. AB - The effects of chronic elevations of endoneurial fluid pressure (EFP) on mammalian nerve fibers were studied using a modified model of experimental galactose neuropathy. Fiber pathology was monitored by the sensitive method of grading the pathologic abnormalities of single teased fibers. Prominent edema was produced in rats poisoned with combined oral and parenteral galactose. EFP and fascicular size were markedly elevated and tibial nerve conduction velocity was reduced. We have demonstrated, for the first time, the presence of fiber degeneration in galactose fed rats. Possible mechanisms of nerve fiber damage include: (1) impaired capillary circulation and increased fiber separation secondary to increased EFP and edema, respectively; (2) increased endoneurial hyperosmolarity, which is known to cause changes in fiber shape; and (3) unknown metabolic derangement of axons. Because much higher rates and different types of fiber pathology are encountered in lead and hexachlorophene intoxication having comparable degrees of EFP, one cannot attribute the fiber pathology directly to the raised EFP in these latter neuropathies. PMID- 6280042 TI - Normal function in sarcoplasmic reticulum from mice with muscular dystrophy. AB - A rapid, gentle technique has been developed for the isolation of sarcoplasmic reticulum (SR) from small amounts of skeletal muscle from normal and dystrophic mice. Assays for mitochondrial and surface membrane marker enzymes revealed only low levels of contamination in the isolated fractions. A small amount of calcium insensitive ("basal") ATPase in the normal preparation and a higher value in the dystrophic were shown to be due to contamination by a lighter membrane fraction of probable surface membrane origin. Isolated SR from normal and dystrophic mice were indistinguishable by thin section and freeze-fracture electron microscopy. Only small differences in calcium loading rates and capacity, and in calcium stimulated ATPase activity, were present. These were attributable to small differences in purity. We conclude that there is no difference in SR from normal and dystrophic mice in the properties measured. PMID- 6280044 TI - Giant axonal neuropathy: an inborn error of organization of intermediate filaments. AB - Light microscopic examination of cultured skin fibroblasts from three patients with giant axonal neuropathy (GAN) revealed large aggregates of intermediate filaments (IF). However, electrophoretic analysis of these cells showed normal amounts of vimentin, the fibroblast IF subunit protein. The isoelectric point and peptide mapping of vimentin were identical to controls. Moreover, vimentin was synthesized and phosphorylated at a normal rate. These findings demonstrate that GAN is an inborn error of the organization of IF and suggest that the basic genetic defect in GAN does not involve primarily the IF subunit proteins. PMID- 6280045 TI - [Report on two cases of mandibular osteomyelitis caused by Candida albicans (author's transl)]. PMID- 6280046 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 18-1982. A 47-year-old man with fever and a mediastinal mass. PMID- 6280048 TI - Long-term follow-up after SV40 inoculation. PMID- 6280049 TI - Evoked potentials in clinical medicine (second of two parts). PMID- 6280047 TI - Calcium homeostasis in immobilization: an example of resorptive hypercalciuria. AB - Prolonged immobilization may result in hypercalcemia, hypercalciuria, and osteoporosis. Although bone resorption is central to this syndrome, the mechanism of resorption is uncertain. In particular, the role of systemic calcium regulating hormones remains unclear. In 14 immobilized subjects we measured fasting calcium excretion, 24-hour urinary calcium excretion during restricted calcium intake, the renal phosphorus threshold, plasma 1,25-dihydroxyvitamin D, nephrogenous cyclic AMP, and immunoreactive parathyroid hormone. Mean serum calcium levels were normal, but fasting and 24-hour calcium excretion were markedly elevated (0.28 mg per deciliter of glomerular filtrate and 314 mg per 24 hours, respectively). The mean levels of serum phosphorus (4.8 mg per deciliter) and the renal phosphorus threshold (4.3 mg per deciliter) were elevated. Mean plasma 1,25-dihydroxyvitamin D was strikingly reduced (9.9 pg per milliliter), as were nephrogenous cyclic (0.64 nmol per deciliter of glomerular filtrate) and immunoreactive parathyroid hormone in both assays. These findings indicate that the parathyroid--1,25-dihydroxyvitamin D axis is suppressed in patients with immobilization-induced hypercalciuria, as would be predicted by a model of resorptive hypercalciuria. PMID- 6280050 TI - Case 53-1981: Cushing's syndrome. PMID- 6280051 TI - ACTH levels after the dexamethasone suppression test in depression. PMID- 6280053 TI - Peptide hormone receptors. PMID- 6280052 TI - A trial of topical acyclovir in genital herpes simplex virus infections. AB - Seventy-seven patients with first episodes of genital herpes and 111 with recurrent episodes were enrolled in a double-blind trial comparing topical acyclovir with a placebo (polyethylene glycol ointment). Among acyclovir-treated patients with first-episode primary genital herpes, the mean duration of viral shedding (4.1 days) and the time to complete crusting of lesions present at the initiation of therapy (7.1 days) were shorter than among placebo recipients (7.0 and 10.5 days, respectively) (P less than 0.05). Acyclovir-treated patients with recurrent herpes had a shorter duration of viral shedding than placebo recipients (0.95 vs. 1.90 days) (P = 0.03). Among the patients with recurrent herpes, acyclovir reduced the time to crusting of lesions in men but had no effect on the symptoms or healing times in women. Topical acyclovir shortens the duration of viral shedding and accelerates healing of some genital herpes simplex virus infections. PMID- 6280054 TI - Long-term complications of virus-induced diabetes mellitus in mice. PMID- 6280055 TI - Unequal crossing-over accounts for the organization of Drosophila virilis rDNA insertions and the integrity of flanking 28S gene. PMID- 6280056 TI - The role of protein phosphorylation in neural and hormonal control of cellular activity. AB - Protein phosphorylation is now recognized to be the major general mechanism by which intracellular events in mammalian tissues are controlled by external physiological stimuli. However, only recently has the idea that different cellular functions are controlled by common protein kinases and protein phosphatases started to gain widespread acceptance. Thus there is an integrated network of regulatory pathways, mediated by phosphorylation-dephosphorylation, that allows diverse cellular events to be coordinated by neural and hormonal stimuli. The evidence that supports this concept is reviewed, with emphasis on the role of protein phosphorylation in enzyme regulation. PMID- 6280058 TI - Conversion of a gonadotropin-releasing hormone antagonist to an agonist. PMID- 6280057 TI - Linkage of beta-thalassaemia mutations and beta-globin gene polymorphisms with DNA polymorphisms in human beta-globin gene cluster. PMID- 6280059 TI - Gating currents associated with potassium channel activation. PMID- 6280060 TI - Enhanced transformation of human fibroblasts by origin-defective simian virus 40. PMID- 6280061 TI - Enhanced metastasis of tumours induced by a SV40 small T deletion mutant. PMID- 6280062 TI - Activation of non-expressed bovine papilloma virus genomes by tumour promoters. PMID- 6280063 TI - Activation of transcription of a yeast gene in E. coli by an IS5 element. PMID- 6280064 TI - Properties of the Ca2+-activated K+ channel in one-step inside-out vesicles from human red cell membranes. PMID- 6280065 TI - All-or-none response of the Ca2+-dependent K+ channel in inside-out vesicles. PMID- 6280066 TI - Intracellular Ca2+ activates a fast voltage-sensitive K+ current in vertebrate sympathetic neurones. PMID- 6280067 TI - Control of cyclic nucleotide metabolism by non-cholinergic, non-adrenergic nerves in rat thyroid gland. PMID- 6280068 TI - Transposition of a tandem duplication of yeast mating-type genes. PMID- 6280069 TI - Low doses of ethanol disrupt sensory responses of brain noradrenergic neurones. PMID- 6280070 TI - Differences in the kinetics of rod and cone synaptic transmission. AB - Photoreceptors of the vertebrate retina hyperpolarize in response to light. The hyperpolarization elicited by a brief flash is approximately ten times slower in rods than in cones of the same retina. We have examined the amplification and temporal properties of synaptic transfer of rod and cone signals to a common postsynaptic element, the horizontal cell. We find that the kinetics of signal transfer at these chemical synapses parallels the speed of the light-evoked signals themselves. PMID- 6280071 TI - Virus-induced diabetes mellitus: autoimmunity and polyendocrine disease prevented by immunosuppression. PMID- 6280072 TI - Non-neutralizing monoclonal antibodies can prevent lethal alphavirus encephalitis. PMID- 6280073 TI - Opiate receptor subclasses differ in their conformational requirements. PMID- 6280074 TI - Dopamine modulates a Ca2+-activated potassium conductance in mammalian hippocampal pyramidal cells. PMID- 6280076 TI - The origin of manganese in cigarette smoke and ash. PMID- 6280075 TI - Lipid-soluble toxins thought to be specific for Na+ channels block Ca2+ channels in neuronal cells. PMID- 6280077 TI - [Herpes genitalis in pregnancy]. PMID- 6280078 TI - [Diagnosis of intraductal tumors in abnormal nipple secretions]. PMID- 6280079 TI - Organization of proviral DNA and protein composition of hormonally stimulated C57Bl/10 strain-associated mouse mammary tumor virus. AB - Two continuous lines, BL-MaTU/A1 and BL-MaTU/s6, were established from C57Bl/10 mammary adenocarcinomas induced by DMBA-prolactin-estradiol treatment. Under in vivo stimulation with dexamethasone, insulin, prolactin, and prostaglandin A1, the cells produce detectable amounts of B-type particles with biochemical properties similar to the GR-MuMTV. Analysis of restriction endonuclease generated fragments of cellular DNA revealed identical patterns in the integration sites and internal recognition sites of MuMTV proviral equivalents in the tumor cells and normal organs of C57Bl/10 strain mice. The restriction DNA fragments of C57Bl/10-associated MuMTV proviral DNA are closely related to those of the Balb/c-associated MuMTV. These results indicate the endogenous origin of MuMTV produced in the hormonally stimulated cultures of DMBA-prolactin-estradiol induced C57Bl/10 mammary tumors. PMID- 6280080 TI - Diagnosis of bovine leukosis virus infection by solid phase radioimmunoassay for virus antibody. AB - Solid phase radioimmunoassay for detection of anti-BLV antibodies in sera of infected animals is described. Viral antigens bound to surface of polystyrene are able selectively adsorb the antibodies from serum. Detection of bound antibodies quantitatively was done by 125I-labeled protein A from Staphylococcus aureus. Technical details of the assay are reported. The assay could be used as competitive one as well. Two different methods for detection of anti-BLV antibodies were compared with solid phase RIA. The solid phase RIA is highly sensitive, specific and available for large-scale examination. The assay is recommended for early diagnosis of bovine leukosis. PMID- 6280081 TI - The World Health Organization. Histological typing of lung tumours. AB - The WHO Histological Classification of Lung Tumours, published in 1967, has been revised. The main features are as follows: Squamous cell carcinoma (epidermoid carcinoma) has the same definition as in the original version, i. e., the identification of keratin and/or intercellular bridges by light microscopy. Three degrees of histological differentiation are described. Dysplasia and carcinoma in situ are discussed. Small cell carcinoma is divided into oat-cell carcinoma, an intermediate cell type and a category for oat-cell carcinomas combined with other major typed, Adenocarcinoma includes the acinar, papillary and bronchiolo alveolar forms and the solid carcinomas with mucus formation (previously part of the large cell carcinoma group). Mesothelial tumours are divided into fibrous, epithelial and biphasic subtypes. A number of less common tumours and tumour-like lesions are defined. PMID- 6280083 TI - Fibres, nutrition and the gut. PMID- 6280082 TI - BCG immunotherapy of lung cancer in a district oncology dispensary. I. Study of 860 patients with histologic diagnosis. AB - The investigation refers to lung cancer patients registered in an oncology dispensary from 1965 to 1975. The conventional treatment (surgery, radiotherapy and chemotherapy) has covered 35.9% of all patients, whereas immunotherapy with BCG or with its soluble fraction has been applied in 46.2% of them. Immunotherapy has mainly been applied to epidermoid carcinoma and large-cell carcinoma. Comparing the survival of immunotherapy treated with non-immunotherapy treated patients it was found that in Stage I-II group the difference in favor of immunotherapy was significant in the second year of observation and in Stage III group a significant difference was found in all observation periods (1-5 years). In Stage I-II the 5-year survival rate of immunotherapy treated patients (13.2%) was similar to the survival rate of operated patients (13.7%). Immunotherapy in large cell carcinoma showed significantly better results than immunotherapy in epidermoid carcinoma. Immunotherapy with BCG and F70 was applied as an independent treatment of lung cancer. When applied to patients primarily operated, radiation or chemotherapy treated, the survival was significantly higher than the survival of patients submitted to conventional treatment or to immunotherapy separately. PMID- 6280084 TI - Elevated cerebellar cyclic GMP levels during the deltamethrin-induced motor syndrome. AB - Adult male Porton albino rats received deltamethrin (0.5--40.0 mg/kg IP) or glycerol formal solvent IP. Their behaviour was observed during the subsequent 110 minutes and the incidence and latency of salivation, tremor and incoordination and spontaneous choreiform episodes were noted. Cyclic GMP levels in the cerebellum were determined by radioimmunoassay at various times after deltamethrin administration. The development of the motor syndrome was dose dependent and followed a specific time course. The threshold dose for profuse salivation and tremor and incoordination was 2.5 mg/kg IP, and for spontaneous choreiform episodes, 5.0 mg/kg IP. The latency of tremor and incoordination decreased significantly as the deltamethrin dose was increased. The first significant increase in cyclic GMP levels in the cerebellum was at 70 minutes, which was 10 minutes after the mean latency of tremor and incoordination. The levels increased further at 100 minutes which was approximately 20 minutes after the mean latency of spontaneous choreiform episodes. Deltamethrin did not have a dose-dependent effect on cyclic GMP levels. The results suggest that deltamethrin changes cerebellar cyclic GMP levels indirectly. PMID- 6280085 TI - Poly(adenosine diphosphate ribose) polymerase activity in neuronal and glial nuclei from bovine cerebrum. AB - Two different preparations isolated from beef cerebrum have been used to compare the polyadenosine diphosphate ribose (polyADPR) polymerase activities in neuronal and glial nuclei: (1) nuclear suspensions (with or without DNase I treatment), and (2) 1 M NaCl nuclear extracts (soluble enzyme). The DNAse I treatment of nuclei and the solubilization of polyADPR polymerase by 1 M NaCl enhances the polyADPR polymerase activity. The polyADPR polymerase activity is similar in neuronal and glial nuclear suspensions, while the neuronal soluble enzyme activity is significantly higher than that of the glial soluble enzyme. Evidence is presented that the difference in soluble enzyme activities is not due to the effects of DNA or degrading enzymes. Some activating factor(s) seem to be present in neuronal soluble extracts, while both inhibiting and activating factor(s) seem to be present in glial soluble extracts. PMID- 6280087 TI - Brain transfer RNA. I. Methylation of newly synthesized tRNA. AB - Present study demonstrates differences between the labeling patterns of rat cerebral cortex and cerebellum. Although the initial rates of synthesis and methylation of tRNA from these two regions of brain are different, their final degree of methylation is the same in both the cases. Differences in the rates of incorporation of radioactivities in both the tissues can be explained by the differences in the rates of equilibrium of the labeled precursors with the pools. Increased incorporation of labeled uridine into cerebral cortical tRNA following dibutyryl cAMP treatment probably resulted from higher uptake of the labeled precursor due to permeability changes. PMID- 6280088 TI - Demyelination induced in mice by avirulent Semliki Forest virus. I. Virology and effects on optic nerve. AB - Swiss A2G mice were infected intraperitoneally with an avirulent strain of Semliki Forest virus. The virus titres in the optic nerves, retina and brain were estimated on post-inoculation days 3 to 8. High titres of virus were obtained in the optic nerve, retina and brain. The optic nerves, retina and brain were studied by light microscopy up to post-inoculation day 67. Electron microscope studies wee carried out on the optic nerve on post-inoculation days 11 and 14. Inflammatory, microcystic changes and demyelination seen in the optic nerve were similar to those found in he central nervous system. PMID- 6280086 TI - Distribution of lipid synthesizing enzymes, 2',3'-cyclic nucleotide 3' phosphodiesterase, and myelin proteins in rat forebrain subfractions during development. AB - The distribution of UDP-galactose:ceramide galactosyltransferase (CGalT) was studied in subcellular fractions of rat forebrain during development using zonal centrifugation on linear gradients. Specialized subfractions: SN 1, a microsomal fraction, SN 4, a myelin-related fraction, and purified myelin were also used for this study. For comparison, two microsomal lipid synthesizing enzymes, a myelin specific enzyme, 2',3'-cyclic nucleotide 3'-phosphodiesterase and myelin proteins were measured in the same subfractions. UDP-glucose:ceramide glucosyltransferase and cerebroside sulfotransferase were confined to microsomes. CGalT was localized in microsomes, but also in myelin and myelin-related fractions. The developmental change in distribution of CGalT in adult animals toward myelin containing fractions could indicate that the replacement of galactosylceramide in compact myelin could be carried out in close proximity to compact myelin (mesaxon, paranodal loops) rather than in the distant oligodendrocyte perikaryon. PMID- 6280089 TI - Demyelination induced in mice by avirulent Semliki Forest virus. II. An ultrastructural study of focal demyelination in the brain. AB - Four-week-old mice were infected intraperitoneally with an avirulent strain A7(74) of Semliki Forest virus (SFV) and killed at 4, 10, 15, 21 and 29 days after inoculation. Focal demyelinating lesions were present by 10 days. These were usually accompanied by a mononuclear infiltrate which included lymphocytes possessing characteristic cytoplasmic projections. These latter extended deep into the cytoplasm of adjacent cells, which were usually astrocytes and macrophages. Other features of the focal lesions were expansion of the extracellular space and demyelination which appeared to be fragmentation or lysis rather than stripping of myelin by macrophages. Although healing occurred in some mice after 4 weeks, acute lesions were still found in others of the same age. It was concluded that the demyelination probably had an immunological basis, and interaction between elements of the immune system and glial cells was a factor which inhibited orderly remyelination of the relatively mild lesions resulting from this infection. PMID- 6280090 TI - GABA neuron systems in hypothalamus and the pituitary gland. Immunohistochemical demonstration using antibodies against glutamate decarboxylase. AB - The distribution of gamma-aminobutyric acid (GABA) nerve fibers and cell bodies in the rat hypothalamus and pituitary gland was immunohistochemically examined using antibodies against the GABA-synthesizing enzyme glutamate decarboxylase (GAD). The dense network of GAD-positive nerve fibers was observed to be essentially evenly distributed throughout the hypothalamus. A plexus of GABA terminals was also demonstrated both in the median eminence and with in the posterior and intermediate lobes of the pituitary. Three distinct clusters of magnocellular GABA neurons were discovered in the posterior hypothalamus. In the addition, GAD immunoreactive cell bodies were observed in many other hypothalamic nuclei, such as the arcuate nucleus and in the perifornical region. The results provide a morphological basis by which GABA of hypothalamic origin may regulate the neuroendocrine system. PMID- 6280091 TI - Changes in neuroendocrine control of anterior pituitary function during aging. PMID- 6280092 TI - Triphasic changes in plasma ACTH concentration and brain serotonin synthesis rate following adrenalectomy in rats. AB - Following bilateral adrenalectomy in adult male rats, there occurs a pattern of triphasic change in basal plasma concentration of radioimmunoassayable ACTH. Plasma ACTH is markedly elevated at 2 h, has returned down almost to normal at 20 h and is again markedly elevated 96 h after adrenalectomy. We have examined serotonin (5HT) synthesis rats in several brain regions, anterior hypothalamus, posterior hypothalamus, and brain stem, at these times after adrenalectomy using the accumulation of 5-hydroxytryptophan (5HTP) after inhibition of aromatic L amino acid decarboxylase with m-hydroxybenzylhydrazine. In both anterior hypothalamus and brain stem, decrease 5HT synthesis rates were observed at 2 and 96 h after adrenalectomy, but at 20 h 5HT synthesis rates were normal. This pattern was not observed in the posterior hypothalamus. Thus, we demonstrated inverse correlations between 5HT synthesis rates in anterior hypothalamus and brain stem, but not posterior hypothalamus, and basal plasma ACTH concentration throughout the period of triphasic change following adrenalectomy in adult male rats. Both the adrenalectomy-induced increases in plasma ACTH concentration and the adrenalectomy-induced decreases in brain 5HT synthesis rates were inhibited by treatment with dexamethasone, suggesting that the changes resulted from glucocorticoid withdrawal. The data are consistent with a role of brain 5HT neurons with cell bodies in brain stem and nerve endings in anterior hypothalamus in the regulation of the triphasic changes in plasma ACTH concentration following adrenalectomy in rats. PMID- 6280093 TI - Differential effects of central adrenoceptor agonists on luteinizing hormone release. AB - This study examined the alterations in episodic luteinizing hormone (LH) release in response to third ventricle infusions of various alpha- and beta-adrenoceptor agonists in ovariectomized (OVX) rats as well as the effects of steroid priming with 50 micrograms estradiol benzoate (EB) and 25 mg progesterone (P) on the LH responses to these agonists. Unanesthetized rats with indwelling atrial cannulae were bled at 10-min intervals for 0.5-1.5 h prior to infusion and up to 1.5 h following infusion of equimolar amounts (0.06 or 0.3 mumol in 2 microliters saline adjusted to pH 5.5 and infused slowly over a 2-min period) of norepinephrine (NE), phenylephrine (Phen, alpha 1-agonist), isoproterenol (Iso, beta-agonist) or clonidine (Clon, alpha 2-agonist). In unprimed OVX rats, 0.06 mumol NE induced a significant lengthening (by approximately 121%) of the episodic interval between the peak LH levels and caused a decrease in mean blood LH levels of approximately 24%, which began almost immediately and lasted for approximately 1 h after infusion. When administered in the same manner and dosage, both alpha- and beta-adrenergic agonists were similarly effective in suppressing pulsatile LH release in OVX unprimed rats, with the following rank order being apparent: Clon greater than NE congruent to Phen greater than Iso. However, in OVX-EBP-primed rats, while 0.06 mumol NE significantly stimulated LH release, none of the other adrenoceptor agonists administered at this dosage was effective in altering the low nonpulsatile levels of blood LH characteristic of the steroid-primed animal. Nevertheless, at a concentration 5 times higher (0.3 mumol) Clon and Phen did induce LH surges while Iso, even at this higher dose, was not stimulatory to LH release. These results suggest that the inhibitory action of NE on LH secretion in OVX rats may be exerted via activation of both alpha- and beta-adrenoceptors, whereas primarily alpha-adrenoceptors are responsible for mediating the NE-induced stimulation of LH release in OVX steroid primed animals. PMID- 6280094 TI - Adenohypophyseal hormones in the CSF. AB - The presence of adenohypophyseal hormones in human cerebrospinal fluid (CSF) had been firmly established by radioimmunoassay procedures. The hormones can originate from the adenohypophysis itself and from brain, as well as from peripheral sites of production. These peptide hormones enter the CSF through the choroid plexus or by direct secretion. The hormones entering by direct secretion into the CSF have a higher CSF-blood ratio than the hormones entering through the choroid plexus. Elevated CSF levels of adenohypophyseal hormones are most likely to occur in the presence of hormone-producing pituitary tumors with suprasellar extension. Although the CSF concentrations of adenohypophyseal hormones may merely reflect their blood concentrations, it is more likely that these hormones also have paracrine as well as endocrine effects. PMID- 6280095 TI - Effect of pentoxifylline on alpha- and beta-adrenoceptor sites in cerebral cortex medial basal hypothalamus and pineal gland of the rat. AB - The intraperitoneal injection of the methylxanthine derivative pentoxifylline (3,7-dimethyl-1-(5-oxo-hexyl)-xanthine] brought about, 3 hr later, a significant depression of alpha- and beta-adrenoceptor sites in the cerebral cortex, and of beta-adrenoceptor sites in medial basal hypothalamus and pineal gland, (assessed from the specific binding of radioactive dihydroergocryptine and dihydroalprenolol respectively). The changes in the density of binding sites were not accompanied by significant modifications of the Kd's. Sympathetic denervation of the pineal gland by superior cervical ganglionectomy (SCGx) abolished the changes of beta-adrenoceptor number in the pineal caused by pentoxifylline. The increase of alpha-adrenoceptor sites in the hypothalamus brought about by ganglionectomy was not affected by injection of pentoxifylline. Pentoxifylline did not compete in vitro for radioligand binding to brain membranes. These results suggest that methylxanthines depress brain adrenoceptor sites acutely, probably by down-regulation of receptors following the increase in catecholamine release caused by injection of the drug. PMID- 6280096 TI - Effect of ethanol administration and withdrawal on benzodiazepine receptor binding in the rat brain. AB - Administration of single doses of ethanol or presence of ethanol in the incubation medium does not change 3H-flunitrazepam binding in rat cerebral cortex, hippocampus and cerebellum. In rats made dependent on ethanol by three daily ethanol administrations for six days and sacrifices 1 hr after the last ethanol dose 3H-flunitrazepam binding is decreased in the cerebral cortex and cerebellum. This decrease is present only in membranes preincubated with Triton X 100 and is due to decreased number of binding sites. In rats sacrificed 16 hours after the last chronic ethanol dose the 3H-flunitrazepam binding returns to control levels. PMID- 6280097 TI - Immunoregulatory cell function in peripheral blood leukocytes of patients with intracranial gliomas. AB - Levels of indomethacin-sensitive, glass-adherent, preculture-sensitive, and lymphocyte-mediated immunoregulatory activity were measured in peripheral blood mononuclear cells from 12 patients with intracranial astrocytomas. The levels of regulatory cell function were determined in assays of T cell immunocompetence as judged by phytohemagglutinin (PHA)-induced in vitro lymphocyte DNA synthesis. Of 6 patients studied before surgical exploration and diagnosis, 6 exhibited significantly depressed levels of PHA responsiveness in association with significantly increased levels of regulatory cell function by glass-adherent, preculture-sensitive and/or indomethacin-sensitive cells. Six patients were studied after diagnosis and treatment. Of those, 2 in remission had normal levels of immune function in association with normal levels of regulatory cell activity, whereas 2 of 4 patients who had recurrent disease had significantly depressed T cell function in association with increased glass-adherent, preculture-sensitive and/or indomethacin-sensitive regulatory cell activity. Although some alterations in lymphocyte-medicated suppressor cell activity were seen in 2 patients, those changes could not be correlated with impaired T cell function as measured in the PHA stimulation assay. The changes in regulatory cell function could not be correlated with levels of immune complexes in patient sera. These data suggest that increased levels of regulatory cell function by glass-adherent, preculture sensitive and/or indomethacin-sensitive cells alone or in conjunction with lymphocyte and T cell depletion are a primary determinant of impaired immunocompetence in glioma patients. PMID- 6280098 TI - Extraneural metastasis of cerebellar glioblastoma multiforme. AB - The case of a 7-year-old girl with a cerebellar glioblastoma and extraneural metastases has been presented. The dura mater was left open at the time of tumor resection. Postoperative hydrocephalus caused prolonged bulging of the incision. We believe that an open dura mater facilitates continuous exposure of extrameningeal tissue to glioma cells that are capable of invading local lymphatics and extracerebral veins, enhancing the probability of widespread dissemination. We suggest that every effort should be made to close the posterior fossa dura mater after resection of a malignant tumor. PMID- 6280099 TI - Pseudotumor cerebri following removal of an adrenocorticotropic hormone-secreting pituitary adenoma. PMID- 6280100 TI - Distribution of opiate receptors in the monkey brain: an autoradiographic study. AB - By employing both in vivo and in vitro labeling techniques, opiate receptors were labeled with tritiated diprenorphine in the monkey brain and localized by light microscopic autoradiography. Both methods of labeling gave similar results, allowing a description of discrete areas having opiate binding sites. High concentrations of opiate receptors were found in the substantia gelatinosa of the spinal cord, nucleus tractus solitarius, area postrema, lateral parabrachial nucleus, substantia grisea centralis, several nuclei of the thalamus and hypothalamus, substantia innominata and in the amygdala. In the monkey pituitary, receptors were found in the neurohypophysis. These results correlate well with those found in autoradiographic studies of the rat brain although there are a few notable differences. Many of the opiate receptor distributions can be correlated with anatomical loci of brain functions known to be influenced by administration of opiate compounds. PMID- 6280101 TI - Effects of locus coeruleus stimulation on neuronal activities of dorsal lateral geniculate nucleus and perigeniculate reticular nucleus of the rat. AB - In rats anesthetized with urethane, a stimulating electrode was introduced to the locus coeruleus by observing the antidromic field response to single shock stimulation of the dorsal pathway of noradrenergic axons. Effects of locus coeruleus stimulation were studied on activities of relay neurons and intrinsic interneurons of the dorsal lateral geniculate nucleus and on those of neurons in the perigeniculate reticular nucleus. The intrinsic interneurons and the perigeniculate reticular neurons are believed to exert inhibition upon the relay neurons. The relay neurons were activated by repetitive stimulation of locus coeruleus; spontaneous discharges were increased in rate and the threshold of response to single shock stimulation of the optic nerve was lowered. The activation was rarely seen in rats pretreated with alpha-methyl-p-tyrosine. Iontophoretic application of phentolamine, an alpha-blocker, effectively antagonized the activation, whereas an iontophoretic beta-blocker and cholinergic blockers were virtually ineffective. The activation of the relay neurons was suggested to be due to a direct action of noradrenaline, released by locus coeruleus stimulation. Locus coeruleus stimulation inhibited the interneurons and activated the perigeniculate reticular neurons; spontaneous or light-evoked discharges were suppressed in the interneurons and tonic discharges were elicited in the perigeniculate reticular neurons. These effects of locus coeruleus stimulation were mimicked by noradrenaline applied iontophoretically. Activation of the perigeniculate reticular neurons was antagonized by an iontophoretic alpha blocker but not by a beta-blocker. Two special features emerge from the present results: (1) the locus coeruleus exerts different effects upon the two neuronal constituents of the dorsal lateral geniculate nucleus, excitation of the relay neurons and inhibition of the intrinsic interneurons; (2) a suggestion previously advocated that locus-coeruleus-induced excitation of the lateral geniculate relay neurons would be due to inhibition of inhibitory neurons (disinhibition) does not hold true, at least with respect to the perigeniculate reticular neurons; the latter neurons have been proved to exert a powerful inhibition upon the geniculate relay neurons and they are excited by stimulation of the locus coeruleus. PMID- 6280102 TI - Non-neurogenic mechanism for reserpine-induced release of catecholamines from the adrenal medulla of neonatal rats: possible modulation by opiate receptors. AB - Neonatal rats do not have functional splanchnic nerve connections to the adrenal medulla until approximately one week of postnatal age, yet they are able to respond to some drugs or stresses by releasing adrenal catecholamines. Reserpine (5 mg/kg s.c.) resulted in significant loss (20-40%) of neonatal catecholamines within 4 h; unlike the acute effects of reserpine in the adult, depletion was not prevented by pretreatment with a nicotinic blocking agent, demonstrating that the effect in the neonate is non-neurogenic. Depletion did not result simply from inhibition of catecholamine storage by reserpine, but rather, the non-neurogenic depletion represented net movement of the amines from the storage granules into the extracellular space, as evaluated by the subcellular distribution of catecholamines. These results suggest that the non-neurogenic mechanism represents release of catecholamines. The immature response mechanism disappeared by 11 days of age and was replaced by a completely neurogenic release, demonstrating that the special mechanism is lost within several days of the onset of functional innervation of the adrenal. Experiments also were carried out to test whether endogenous opioids and/or opiate receptors are involved in the non neurogenic mechanism. Naloxone (5 mg/kg s.c.) potentiated the depletion of catecholamines by reserpine, while methadone (2.5 mg/kg s.c.) inhibited the non neurogenic response. In contrast, no potentiation of release by naloxone or inhibition by methadone was seen in adult rats. Thus, opiate receptors may modulate only the immature secretory mechanism. PMID- 6280103 TI - Heterotopic stimuli-related potential gradients in a small volume of the medullary reticular formation in the cat. AB - In anaesthetized cats the role of the medullary reticular formation in discriminative functions was studied. We analysed (1). variability of collective reactions of the reticular neurons (extracellular potential gradients recorded bipolarly in three directions by a tetrad of micropipettes with closely positioned tips in a small volume of the reticular formation) to the same stimulus, and (2). reactions to heterotopic stimuli. The pattern of the potential gradients generated by the stimulation (electrical, natural) of the same spot on the body surface was constant during the time periods studied (up to 30 min). Responses evoked from reciprocally remote receptor areas were manifestly different, at least in one of the depth profiles investigated. It can be concluded that the reticular formation, through its reactions, can differentiate heterotopic stimuli; in this respect it does not operate as a 'nonspecific' system. PMID- 6280104 TI - The relationship between the membrane potential of neurosecretory nerve endings, as measured by a voltage-sensitive dye, and the release of neurohypophysial hormones. AB - The membrane potential of isolated rat neurohypophyses and isolated neurosecretosomes (neurosecretory nerve endings) was monitored with the voltage sensitive fluorescent probe diS-C3-(5). K ions, in contrast to Na or Cl ions, give rise to large changes of the fluorescent signal. The fluorescent response is linearly related to log[K+]0 at values higher than 10 mM, whereas at lower [K+]0 the permeability of the membrane for Na ions has to be taken into account. Veratridine increases the fluorescent signal only in the presence of external sodium; this effect is blocked by tetrodotoxin. After prolonged K-induced depolarisation, addition of veratridine to the medium gives a further change in fluorescence of diS-C3-(5) associated with release of vasopressin. Vasopressin release from isolated neurohypophyses started to increase significantly only above 25 mk [K+]0, while the depolarization of the membrane was linearly related to log[K+]0. The results are consistent with the view that neurosecretory nerve endings have voltage-dependent calcium channels that regulate the amount of hormone released during depolarisation. PMID- 6280105 TI - Peripheral neuropathies and tremor. AB - We studied 14 patients with tremor and acquired peripheral neuropathies of different origin. Minimal weakness was the only clinical finding common to all patients. Sensory changes, etiology, and course of the neuropathy varied. NO unique pattern of peripheral nerve histopathology could be demonstrated. Slowing of conduction velocity of Ia fibers was found, but the role played by impairment of sensory inputs remains unclear. Tremor associated with peripheral neuropathy seems to be due to enhancement of physiologic tremor by minimal weakness. PMID- 6280106 TI - Adrenoleukodystrophy: elevated C26 fatty acid in cultured skin fibroblasts and correlation with disease expression in three generations of a kindred. AB - Varying combination of central and peripheral nervous system disease and Addison disease were previously described in 14 members of a family. The diagnosis of adrenoleukodystrophy (ALD) was documented in affected individuals by increased content of C26:0 C26/C22 fatty acid ratios were not proportional to the neurologic syndrome, severity of disease, or duration. PMID- 6280107 TI - Familial X-linked Addison disease as an expression of adrenoleukodystrophy (ALD): elevated C26 fatty acid in cultured skin fibroblasts. AB - Adrenoleukodystrophy (ALD) is a fatal X-linked disorder of very long chain fatty acid (VLCFA) metabolism manifested by disease of the central and peripheral nervous systems and the adrenals. X-linked Addison disease alone, as an expression of ALD, has not been previously reported. We present the results of our study of a unique family among whom clinically apparent Addison disease without neurologic involvement has occurred in affected males, and spastic paraparesis has occurred in female carriers. The presence of ALD was confirmed by VLCFA determination in cultured skin fibroblasts. A comparison group of patients with Addison disease on a putative immunopathogenic basis was normal. PMID- 6280108 TI - Detection and localization of phosphatases, DNA, glycogen, mucopolysaccharides and bound lipids in the testis of a teleost fish, Anabas testudineus (Bloch) during the annual cyclical changes by histochemical methods. PMID- 6280109 TI - [Infections in dialysis centers. Problems and technics of control]. PMID- 6280110 TI - Benzodiazepines enhance the bicuculline-sensitive part of recurrent Renshaw inhibition in the cat spinal cord. PMID- 6280111 TI - Inhibition of the electrically induced release of [3H]dopamine by serotonin from superfused rat striatal slices. AB - The effect of serotonin (5-hydroxytryptamine) on the electrically induced release of [3H]dopamine from superfused slices of the rat striatum has been studied. It was observed that serotonin produced a concentration dependent decrease in the field stimulation-induced release of [3H]dopamine with the threshold concentration being 10(-6) M or lower. Methysergide, in a concentration which did not alter the evoked release, antagonized the inhibitory effect of serotonin. The present results suggest that serotonin should be added to the list of endogenous substances that can influence dopaminergic transmission in the striatum. PMID- 6280112 TI - Direct projections from the extrathalamic forebrain structures to the neocortex in the macaque monkey. AB - Extrathalamic direct projections from the subcortical forebrain structures to the neocortex were examined in the macaque monkey by the horseradish peroxidase method. The enzyme, when injected into discrete regions in the neocortex, labeled cell bodies of extrathalamic forebrain neurons in the basal nucleus of Meynert, nucleus of the diagonal band, medial septal nucleus, hypothalamus, claustrum and dorsolateral part of the basal amygdaloid nucleus. Neurons in the basal nucleus of Meynert, lateral hypothalamus and claustrum appeared to send their axons widely, but not diffusely, to the neocortex. PMID- 6280113 TI - Residual cerebellothalamic terminal fields following hemispherectomy in the cat. AB - Tritiated amino acids were injected into the nucleus interpositus, both the anterior (INA) and posterior (INP) divisions, and the projections to the ventrolateral nucleus (VL) of the thalamus mapped autoradiographically in 7 intact and 8 hemispherectomized adult cats. Five hemispherectomized cats receiving injections into the antero-medial one-third of INP, exhibited terminal fields in lateral portions of the contralateral VL. These terminals were patchy and appeared qualitatively identical to those seen in intact animals. In 3 hemispherectomized cats sparse terminal fields were also observed in both normal and gliotic regions of the parafascicular, central lateral, anterior pretectal and posterior thalamic nuclei. The presence of these terminal fields was specific to the injections into the INP. The density of the terminal labeling was greater in the hemispherectomized than in the intact cats. The VL areas receiving these terminals were conspicuously gliotic and devoid of large thalamocortical relay neurons. These results indicate that the INP projections to the VL persist despite degeneration of the thalamocortical neurons, suggesting that the site of the residual terminals is not directly upon such neurons. Attempts at reorganization are discussed. PMID- 6280114 TI - An excitatory input to nucleus raphe magnus from the red nucleus in the cat. AB - In chloralose-anaesthetized cats, with the cerebellum removed, stimulation in the red nucleus excited the majority (60-65%) of neurones in nucleus raphe magnus (NRM), including raphespinal neurones. Evidence was obtained for both monosynaptic and polysynaptic excitation. The projection was confirmed by recording antidromic responses in the red nucleus to stimulation in NRM. It is suggested that the role of NRM in motor control is to inhibit spinal flexion responses to peripheral stimuli so that commands from the red nucleus and other motor control regions may take place without interruption. PMID- 6280115 TI - Do opioid peptides mediate a presynaptic control of C-fibre transmission in the rat spinal cord? AB - A presynaptic inhibitory role for opioid peptides in the control of C-fibre evoked activity in the dorsal horn has been investigated. The excitability of C fibre terminals in the dorsal horn of decerebrate spinal rats was tested using intraspinal terminal stimulation and recording the size of the antidromic C wave from the dorsal roots. Naloxone (1-2 mg/kg) failed to alter the baseline terminal excitability of the C-fibres, but reduced the increase in terminal excitability produced by A-fibre afferent conditioning stimuli. The inhibition of postsynaptic C-evoked activity in lamina 5 cells produced by A-afferent fibre conditioning stimuli was also reduced by naloxone. This effect may reflect the reversal of an opioid-mediated presynaptic inhibition, although blockade of a direct postsynaptic inhibitory action could also be involved. PMID- 6280116 TI - Cerebellar cortical afferents from the dorsal column nuclei in sheep, demonstrated with retrograde transport of horseradish peroxidase. AB - Following horseradish peroxidase injections in the cortex of the sheep cerebellum (except the ventral part of the anterior lobe, the flocculus and ventral paraflocculus), labeled cells were found in nucleus gracilis, medial cuneate and lateral cuneate. The present findings provide evidence that the projection of the dorsal column nuclei to the cerebellum in the sheep is more extensive than has been reported for the common laboratory animals. Projections to the vermis are bilateral, projections to the hemisphere are mainly ipsilateral. PMID- 6280117 TI - Aggression induced by intermittent positive reinforcement. AB - Mammalian and non-mammalian species engage in aggressive behavior toward animate and inanimate targets when exposed to intermittent access to a positive reinforcer. This behavior, called extinction- or schedule-induced aggression, typically includes a biting or striking topography that inflicts damage on a target. This paper critically reviews research and theoretical issues concerning such aggression and suggests directions for future investigation. PMID- 6280118 TI - Schedule-induced self-injection of drugs. AB - Studies of acquisition and maintenance patterns of drug intake, including dependence, necessitate voluntary intake of drugs. Voluntary intake in animal studies is difficult to achieve because of aversive taste factors associated with most drugs, and involuntary or forced choice methods obscure the behavior which is the object of study. The schedule-induced polydipsia paradigm has been used to induce oral ingestion of large volumes of alcohol, barbiturate and other drug solutions. We have developed a method of schedule-induced self-injection which allows the study of acquisition and maintenance of drug intake behavior with changing environments free from the interference of taste factors or imbalances due to excessive water intake. In this paper we review our findings on the acquisition and maintenance patterns of amphetamine, methadone, heroin, alcohol, nicotine, cocaine, delta 9-THC and haloperidol. For all drugs except amphetamine, the combination of schedule and nutritional deprivation leads to the highest rates of drug intake as compared to controls. The schedule does not appear to be a potent factor at 90% and free feeding weight, and drug intake is the result of interaction of environmental factors and pharmacological properties of the drugs, rather than the effects of drug or environmental factors separately. The maintenance patterns of nicotine, cocaine, heroin and alcohol are also discussed and the advantages of schedule-induced self-injection over schedule-induced polydipsia methods are presented. PMID- 6280119 TI - [Age-related changes of cAMP and prostaglandin content in the brain and lymphocytes of spontaneously tumor-producing C3H mice]. AB - Age caused concurrent changes in the activation of the adenylate-cyclase activity by dopamine in the diencephalon and the stimulation of the basal level of cAMP by isoproterenol in the lymphocytes were demonstrated in the spontaneously tumor producing C3H mice. These changes may be related to the occurrence of tumors in these animals. The above-mentioned changes in the cAMP system were not found in AB mice. By treating the adult C3H mice with substances which increase the cAMP level, it was possible to achieve a renewed stimulation of the cAMP system as seen in young mice. With this treatment the spontaneous tumor-induction rate was also reduced. In the adenocarcinomas of the C3H mice a decrease in the level of prostaglandin E and an increase in the level of prostaglandin F2 alpha was observed. PMID- 6280120 TI - [So-called histiocytosis X and malignant histiocytosis]. AB - A retrospective study of 12 cases of so called histiocytosis-X and 3 cases of malignant histiocytosis was done. It was possible to establish the differences in clinical, morphologic and cytochemical findings of both diseases. The diagnosis of histiocytosis-X can be confirmed by multinucleated histiocytes interrupted by eosinophils and plasmacells. The histopathology of malignant histiocytosis is different and is characterized by atypical histiocytes. Erythrophagocytosis throughout the tissues is seen. Typical histochemistry (acid phosphatase and naphtol-AS-acetat-esterase) findings are also helpful for diagnosis. The treatment of both diseases should be continued at least six months after disappearance of clinical apparent lesions. Combination chemotherapy with vinblastine and prednisone is suggested. In cases of histiocytosis-X in isolated lesions curettage or irradiation may be adequate. Long term remissions and presumed cures of histiocytosis-X are possible in over 70% of the cases. A strict correlation between the prognosis and the degree of involvement is confirmed. Even in cases of malignant histiocytosis, previously reported as rapidly fatal disease, combination chemotherapy may produce complete long term remissions. PMID- 6280121 TI - Decision-making in the therapy of external eye disease: noninfected corneal ulcers. AB - Despite diverse and multiple clinical etiologies, the pathogenesis of noninfected corneal ulcers consistently relates to persistent or extensive epithelial defects, concomitant with stromal inflammatory cell infiltration and enzymatic degradation of collagen and ground substance. The therapeutic approaches to these disorders may be considered on three levels: (1) the determination of etiology and initiation of primary therapy; (2) promotion of epithelial healing; and (3) limitation of ulceration and support of repair, the latter aspects currently featuring pharmacologic management and limited surgical modalities, particularly the use of tissue adhesive. PMID- 6280123 TI - [The nasopharyngeal carcinoma and Epstein-Barr virus (EBV) specific IgA (author's transl)]. PMID- 6280122 TI - A comparison of membrane-bound enzymes of the isolated brush border plasma membranes of the cestodes of Hymenolepis diminuta and H. microstoma. AB - Preparations of isolated brush border plasma membrane of Hymenolepis diminuta and H. microstoma possess the following enzymatic activities: alkaline phosphohydrolase (E.C. 3.1.3.1); Type I phosphodiesterase (E.E. 3.1.4.1); ribonuclease (E.C. 3.1.4.22); adenosine triphosphatase (E.C. 3.6.1.3); and 5' nucleotidase (E.C. 3.1.3.5). The following enzymatic activities could not be demonstrated in either membrane preparation: Type II phosphodiesterase (E.C. 3.1.4.18); cyclic adenosine-3', 5'-monophosphate phosphodiesterase (E.C. 3.1.4.17); leucine aminopeptidase (E.C. 3.4.11.1); maltase (alpha-glucosidase; E.C. 3.2.1.20); and lactase (beta-galactosidase; E.C. 3.2.1.23). These data generally agree with those of previous studies in which similar membrane-bound enzymes were demonstrated in intact (living) worms. PMID- 6280124 TI - [Anterior pituitary and adrenocortical hormones and cellular immunity in newborn infants with staphylococcal infection]. PMID- 6280125 TI - Analysis of creams. II. Quantitative determination of drugs in creams by titration in non-aqueous solvents. AB - The possibilities for the determination of active components in creams by acid base titrations in non-aqueous solvents were investigated. Interference by cream base components with the titration of weak organic bases and their halides with perchloric acid in acetic acid, and with the titration of weak acids with tetrabutylammonium hydroxide in N,N-dimethylformamide were studied. It appeared to be possible to determine alkaloid halides, salicylic acid, hexachlorophene and methyl salicylate without previous clean-up of the cream samples. PMID- 6280126 TI - The effect of intracellular cyclic nucleotides and calcium on the action potential and acetylcholine response of isolated cardiac cells. AB - Ventricular and atrioventricular nodal cells from guinea pig and rabbit hearts were isolated by perfusing the heart with collagenase (Langendorff perfusion). In these cells the cyclic nucleotides cAMP and cGMP or Ca and EGTA were injected through a microelectrode by pressure (0.5-3 kg/cm2). The effect of injection on both the action potential and the hyperpolarization induced by acetylcholine was studied. The following results were obtained. 1. cAMP prolonged the ventricular action potential and shifted the plateau to more positive potentials. The configuration of the A-V nodal action potential was not detectably changed by cAMP injection, but the spontaneous rate was increased. 2. cGMP first shortened the ventricular action potential. In most experiments this effect was followed by long lasting prolongation of the action potential. 3. Both extracellular and intracellular application of dibutyryl cGMP shortened the ventricular action potential but did not produce a subsequent prolongation. However, prolongation was observed on injection of GMP, the direct metabolite. 4. Injection of cGMP in nodal cells did not hyperpolarize the membrane nor slow the spontaneous rate; rather, an increase in rate was observed. 5. The acetylcholine-induced hyperpolarization was not altered in amplitude or time course by the injection of cAMP, cGMP, Ca or EGTA. 6. The results support the hypothesis that cGMP might be involved in the control of voltage-controlled ionic channels but suggest that it does not play a role as a mediator of the classical muscarinic action i.e. the activation of a specific potassium channel by acetylcholine. PMID- 6280127 TI - Facilitation and impulse propagation failure at the frog neuromuscular junction. AB - Exposure of frog neuromuscular junctions to solutions which contain a high concentration of calcium ions produces failure of neuromuscular transmission. This failure of transmission is abrupt and usually complete. However, some terminals produce small end-plate potentials even after the exposure to a high concentration of calcium ions. A second stimulus to the nerve can overcome the block of neuromuscular transmission if the interval between the stimuli is less than a critical value. The size of the end-plate potential is almost independent of the interstimulus interval if the latter is less than the critical value but more than the refractory period. The depth of this neuromuscular block is affected by temperature, potassium ions, osmotic pressure, cobalt ions, and prior high frequency stimulation of the nerve. Neuromuscular transmission failure coincides with failure of the nerve action potential (NAP) to invade the terminal. Prior to propagation failure, the second extracellularly recorded NAP is smaller, but is conducted faster than the first NAP. The relevance of these findings to the facilitation of transmitter release seen in solutions of normal divalent ion content is discussed. PMID- 6280128 TI - A quantitative analysis of the Na+-dependence of Vmax of the fast action potential in mammalian ventricular myocardium. Saturation characteristics and the modulation of a drug-induced INa blockade by [Na+]o. PMID- 6280129 TI - [Transcatheter hepatic artery embolization in 60 patients with hepatocellular carcinoma studies on angiographic features (author's transl)]. PMID- 6280130 TI - [Mediation by opioid peptides of anterior pituitary response to insulin-induced hypoglycemia (author's transl)]. AB - A 1.6 mg dose of the specific opiate antagonist naloxone was administered to normal volunteers, either separately or in conjunction with insulin-induce hypoglycemia, in order to study the possible mediation by opioid peptides of the release of adrenocorticotrophin (ACTH), lipotropins (LPH), human growth hormone (GH) and prolactin (PRL). Administered separately, naloxone was associated with a significant fall in PRL levels (p less than 0.01), a significant and unexpected rise in GH levels (p less than 0.02), and a suppression of the circadian decrease of ACTH and LPH levels. The association of naloxone with insulin-induced hypoglycemia significantly reduced the PRL peak (p less than 0.05), did not affect the rise of GH and lowered the ACTH peak, without altering the LPH peak. These data suggest the existence of a positive opioid tone to PRL secretion, as well as an opioid peptide role in the PRL response to hypoglycemia. They argue against the likelihood of an opioid pathway in the GH response to hypoglycemia. Furthermore, the data favor a paradoxical effect of naloxone on ACTH release during insulin-induced hypoglycemia. PMID- 6280131 TI - [The Verner-Morrison syndrome: endocrine cholera or vipoma? (author's transl)]. AB - This syndrome, also known under the initials W.D.H.A., is due to a single or multiple pancreatic tumour or to micropolyadenomatosis consisting of non-beta islet cells. Malignancy is found in about two-thirds of the cases. The other endocrine glands are rarely involved. The syndrome is more frequent in women than in men. It is characterized by liquid diarrhoea, marked hypokalaemia and absence of gastric hyperacidity. The tumour is mainly diagnosed by echotomography, computerized tomography and arteriography. It can also be located by staged collections of blood along the portal system for hormonal assays. The nature of the tumour can only be ascertained by demonstrating the presence of the responsible hormone, usually the "vaso-intestinal peptide". Treatment is primarily surgical. Adjuvant treatments include streptozotocine and embolization by superselective catherization in cases of hepatic matastases. The prognosis is sombre since in spite of the various treatments cure can only be achieved in 50% of the patients. PMID- 6280132 TI - [Haemophilus influenza infections in infants and mothers. Three cases (author's transl)]. AB - The three cases reported were diagnosed at the time of contamination by direct examination and culture, at birth, of placental tissue, gastric fluid and peripheral specimens. The culture media included a chocolate agar medium favourable to the growth of H. influenzae. One of the isolates was beta-lactamase producer and therefore resistant to the group A penicillins usually prescribed. Determination of the M.I.C.s of eight antibiotics showed that cefotaxime constitutes, for the time being, a suitable alternative to penicillins against such strains. Early detection of H. influenzae perinatal infections make it possible to treat neonates before complications develop. Among the 19 cases published, there were 4 cases of meningitis, 8 cases of septicaemia and 1 case of arthritis. PMID- 6280133 TI - Platelets in primary thrombocythemia: electron microscopic study. AB - The ultrastructural analysis of platelets from patients with Primary Thrombocythemia (PT) showed some anomalous features. Platelets having structural modifications of their organelles were observed and a decrease in the number of dense bodies was seen. Freeze-fracture analysis of the plasma membrane of platelets from PT patients showed a significant increase in the number of intramembranous particles (IMP). PMID- 6280134 TI - Construction and partial characterization of two recombinant cDNA clones for procollagen from chicken cartilage. AB - Type II procollagen mRNA has been partially purified from embryonic chick sternal cartilage by guanidine hydrochloride extraction, sucrose gradient sedimentation and Sepharose 4B chromatography. Double stranded cDNA was synthesized using AMV reverse transcriptase and E. coli DNA polymerase I, tailed using terminal transferase and inserted into the Pst I site of pBR322. Two putative type II procollagen cDNA clones have been characterized. Both plasmids hybridize to 2 sternal RNA species, a major species of 5.3 kb and a minor species of 7 kb. These RNAs are present in total RNA from sterna and differentiated limb bud cultures but are not detected in RNA from stage 24 limb bud which has not yet differentiated to cartilage or in RNA from calvaria. The time of appearance of these RNAs during the differentiation of limb mesenchyme in culture parallels the appearance of translatable type II procollagen mRNA. PMID- 6280136 TI - Gene shuttling: moving of cloned DNA into and out of eukaryotic cells. AB - Successful shuttling of cloned DNA in eukaryotic cells should allow isolation of expressed genes. We tested the utility of cosmids for moving DNA into and out of eukaryotic cells. The unique cleavage of DNA at the cos site by the terminase function of lambda was exploited to maintain the linkage between the vector and inserted gene sequences, a prerequisite for successful rescue of the transforming DNA from high molecular weight DNA of the eukaryotic transformant. A cosmid recombinant containing the HSV thymidine kinase gene and a lambda recombinant containing the chicken thymidine kinase gene were used to test the feasability of this method. It was found that these recombinants can be rescued with high efficiency from DNA of HAT-resistant cells. PMID- 6280135 TI - Characterization of sequence elements at the 5' end of a discoidin I gene isolated from Dictyostelium discoideum. AB - The discoidin I genes of Dictyostelium discoideum encode a family of three closely related developmentally regulated lectins which may play a role in cell cohesion. We have isolated a genomic clone containing the 5' half of a discoidin I gene with 12kb of flanking sequence and we have identified the discoidin I polypeptide encoded by this gene. We have determined part of the nucleotide sequence of the cloned segment and have hence determined the N-terminal amino acid sequence of the encoded polypeptide. The nucleotide sequence upstream of the AUG initiation codon was unusually AT rich, with 94% AT base pairs in the 171 nucleotides for which sequence was determined. However, having determined the start point of transcription ('cap' site) two striking features of similarity with other eukaryotic structural genes became apparent. The sequence TATAAA was present 33 nucleotides upstream from the 'cap' site and the sequence CCAAT was present a further 20 nucleotides upstream. PMID- 6280137 TI - Sequence analysis of bovine satellite I DNA (1.715 gm/cm3). AB - The 1402 bp Eco RI repeating unit of bovine satellite I DNA (rho CsCl = 1.715 gm/cm3) has been cloned in pBR322. The sequence of this cloned repeat has been determined and is greater than 97% homologous to the sequence reported for another clone of satellite I (48) and for uncloned satellite I DNA (49). The internal sequence structure of the Eco RI repeat contains imperfect direct and inverted repeats of a variety of lengths and frequencies. The most outstanding repeat structures center on the hexanucleotide CTCGAG which, at a stringency of greater than 80% sequence homology, occurs at 26 locations within the RI repeat. Two of these 6 bp units are found within the 31 bp consensus sequence of a repeating structure which spans the entire length of the 1402 bp repeat (49). The 31 bp consensus sequence contains an internal dodecanucleotide repeat, as do the consensus sequences of the repeat units determined for 3 other bovine satellite DNAs (rho CsCl = 1.706, 1.711a, 1.720 gm/cm3). Based on this evidence, we present a model for the evolutionary relationship between satellite I and the other bovine satellites. PMID- 6280138 TI - Five nucleotide changes in the large intervening sequence of a beta globin gene in a beta+ thalassemia patient. AB - A beta globin gene from a patient with homozygous beta+ thalassemia has been cloned and completely sequenced. No changes from normal are found in the 200 nucleotides 5' to the cap site, in the 3' untranslated region up to the poly A addition site, in the small intervening sequence (IVS 1), or in the coding sequence except for a third base change in codon 2. The only other differences are in the large intervening sequence (IVS 2). One of these, at a position 16 nucleotides from the 5' end of IVS 2, has been reported previously in normal individuals, and is probably a polymorphism. Four other changes, at positions 74, 81, 666, and 705 are also seen in IVS 2. Abnormal beta globin mRNA precursors detected in the bone marrow cells of this patient, and abnormal beta globin RNA splicing observed when this gene is transcribed in a tissue culture system taken together with these IVS 2 changes, suggest that the beta+ thalassemia phenotype is produced by a decrease in normal beta globin mRNA processing. PMID- 6280139 TI - Coordinate regulation of the four tubulin genes of Chlamydomonas reinhardi. AB - During cell division and during the induction of tubulin synthesis that accompanies flagellar regeneration in Chlamydomonas reinhardi, four tubulin mRNAs of discrete molecular sizes are produced. During induction two beta tubulin mRNAs (2.47 kb and 2.34 kb) and two alpha tubulin mRNAs (2.26 kb and 2.13 kb) are synthesized in high abundance and in a closely coordinated fashion. Combined data from restriction enzyme mapping (i.e., Southern analysis) of genomic DNA and of Charon 30 recombinant clones bearing inserts of Chlamydomonas tubulin genes provide direct evidence for four distinct tubulin genes in this organism. Dot blot analysis of the level of hybridization of a 32p nick-translated beta tubulin cDNA to genomic DNA from gametic cells and to a clone containing the beta 1 tubulin gene indicate that each beta 1 tubulin gene is present in one copy per cell. Additional hybridization experiments employing fragments of cDNA clones which selectively anneal to either the 3' or 5' portions of the two alpha tubulin genes or to one or both of the two beta tubulin genes suggest that each tubulin gene is actively transcribed to give rise to one of the four tubulin mRNAs. These observations further suggest that at most four basic types of tubulin subunits are produced by Chlamydomonas and that the heterogeneity of tubulin subunits reported to exist in the flagellar axoneme must arise as a result of post translational modification. PMID- 6280140 TI - Molecular cloning and nucleotide sequencing of the gene for E. coli cAMP receptor protein. AB - The crp gene of E. coli, which codes for cAMP receptor protein (CRP), has been cloned in the plasmid pBR322 on the basis of a genetic complementation. One of the recombinant plasmids, pHA1, was shown to direct the synthesis of CRP in a cell-free system. The location of the crp gene was determined by constructing subclones carrying various portions of pHA1. The nucleotide sequence of the crp gene has been determined. The coding region consists of 627 base pairs (bp), which specify a protein of 209 amino acids. The predicted amino acid sequence from the DNA sequence is consistent with the amino acid sequence partially known and the amino acid composition of CRP. After the coding region, there is a G-C rich inverted repeat sequence followed by a run of Ts, which could be a terminator of the crp gene. A possible promoter sequence was found about 180 bp upstream from the initiation codon and was shown to act as a promoter in vitro and in vivo. There are two dyad symmetry regions in a 167 bp leader sequence. PMID- 6280141 TI - Cloning and sequence of the crp gene of Escherichia coli K 12. AB - We have determined the nucleotide sequence of the crp gene of Escherichia coli K 12. From a lambda transducing phage, the crp region was subcloned into pBR322. The gene was localized on the cloned fragment by determining the length of deletions which affect its expression. Its nucleotide sequence was established by using the technique of Maxam and Gilbert. The deduced amino-acid sequence is in agreement with the previously published amino acid composition of the protein (1, 2). Analysis of the sequence confirms that the DNA binding domain is located in the C-terminal portion of the protein. PMID- 6280142 TI - Poly(adenylic acids) containing the antibiotic tubercidin -- base pairing and hydrolysis by nuclease S1. AB - Poly(adenylic acids) containing the antibiotic tubercidin (7-deazaadenosine) form double strands with poly(uridylic acid) by Watson-Crick base pairing. The stability of these complexes is enhanced by an increasing adenosine content of the polymers. Whereas poly(tubercidylic acid) can bind only one poly(U) chain, the copolymers of adenylic and tubercidylic acid bind a second strand of poly(U). The melting temperatures imply a triple strand formation in a similar geometry as found for poly(A).2poly(U). The diminished hypochromicity of those complexes suggests semi-Hoogsteen base pairs, caused by the lack of N-7 in the antibiotic. As found for poly(A).poly(U), the double-stranded poly(Tu).poly(U) is not hydrolyzed by nuclease S1. In contrast to the four regular homopolyribonucleotides the single-stranded poly(Tu) is cleaved very rapidly. This may be due to a great flexibility of the polynucleotide chain. Moreover TuMP does not inhibit the enzymic digestion. Both phenomena imply a mechanism for the antibiotic action of tubercidin on the polymer level. PMID- 6280144 TI - Characterization of two new plasmid DNAs found in mitochondria of wild-type Neurospora intermedia strains. AB - Mitochondria from two Neurospora intermedia strains (P4O5-Labelle and Fiji N6-6) were found to contain plasmid DNAs in addition to the standard mitochondrial DNA species. The plasmid DNAs consist of monomeric circles (4.1-4.3 kbp and 5.2-5.3 kbp for Labelle and Fiji, respectively) and oligomers in which monomers are organized as head-to-tail repeats. DNA-DNA hybridization experiments showed that the plasmids have no substantial sequence homology to mtDNA, to each other, or to a previously characterized mitochondrial plasmid from N. crassa strain Mauriceville-lc (Collins et al. Cell 24, 443-452, 1981). The intramitochondrial location of the plasmids was established by cell fractionation and nuclease protection experiments. In sexual crosses, the plasmids showed strict maternal inheritance, the same as Neurospora mitochondrial DNA. The plasmids may represent a novel class of mitochondrial genetic elements. PMID- 6280143 TI - Restriction and modification enzymes and their recognition sequences. PMID- 6280145 TI - Structural analysis of repetitive DNA sequences in the bovine corticotropin-beta lipotropin precursor gene region. AB - Repetitive DNA sequences in the bovine corticotropin-beta-lipotropin precursor gene region have been mapped and subjected to nucleotide sequence analysis. Two of the four repetitive DNA segments found are located in the 5'-flanking region, and one each within the intervening sequences. Each repetitive DNA segment contains one to three highly homologous unit sequences with an approximate length of 120 base pairs. All the unit sequences are flanked on the 3' side by tandem repeats. There are about 10(5) copies of the repetitive DNA in the bovine genome. Comparison of the bovine repetitive sequences with those of other mammalian species reveals the presence of a homologous segment of approximately 40 base pairs. This segment and the region preceding it in the bovine repetitive DNA exhibit sequence homology with the region encompassing the origin of DNA replication in papovaviruses. PMID- 6280146 TI - Repetitive DNA sequences in the human corticotropin-beta-lipotrophin precursor gene region: Alu family members. AB - Repetitive DNA sequences in the human corticotropin-beta-lipotropin precursor gene region have been studied by blot hybridization analysis and DNA sequencing. Six repetitive sequences are present in this gene region; five of them are Alu family members with an approximate length of 300 base pairs, and the other consists of a portion of an Alu family sequence. Two of these Alu family members are located in the 5'-flanking region of the gene, and the remaining four within the intervening sequences. These Alu family sequences constitute inverted repeats in the intervening sequences as well as in the 5'-flanking region of the gene. PMID- 6280147 TI - The nucleotide sequence at the transcription termination site of the ribosomal RNA gene in Tetrahymena thermophila. AB - The sequence of 415 nucleotides surrounding the transcription termination site for ribosomal RNA in Tetrahymena thermophila has been determined. The positions of the 3'-ends of mature 26S rRNA, pre-26S rRNA and 35S pre-rRNA were localized within this sequence by hybridization of the purified RNA species to be selected DNA fragments, followed by S1 nuclease treatment of the hybrid and a precise sizing of the RNA-protected DNA fragments on sequencing gels. The 35S pre-rRNA population contained molecules with two distinct 3'-ends, one of which is identical to the end of pre-26S and 26S rRNA, while the other corresponds to a position 15 nucleotides further downstream, which is assumed to be the transcription termination site. The non-coding DNA strand contains a cluster of T's at the putative termination site, and several other T clusters are found further downstream. A short inverted repeat sequence is located near the putative termination site within the transcribed region. The possible role of these structures for transcription termination is discussed. PMID- 6280148 TI - Comparative analysis of Xenopus tropicalis and Xenopus laevis vitellogenin gene sequences. AB - Analysis of cDNA clones synthesized from vitellogenin mRNA of X. tropicalis revealed three different types of cDNA clones, i.e. A, A* and B. A and A* clones have a sequence divergence of about 6% and are both related to X. laevis vitellogenin cDNAs of subgroup A1 as well as A2 with a sequence divergence of 6 9%. B clones however, are related to X. laevis cDNA clones of subgroup B1 and B2 with a sequence divergence of about 7%. While the A and B clones correspond to vitellogenin mRNAs of similar abundance, A* clone is complementary to a vitellogenin mRNA about 100 fold less abundant than A and B mRNAs although all three vitellogenin mRNAs are encoded by single copy genes. Furthermore, two forms of A* mRNA were found. One of the two is lacking an internal fragment of about 900 bp. Since this DNA fragment is highly repeated in the genome, we suggest that this A* clone was synthesized from a processing intermediate of the A* precursor vitellogenin mRNA. PMID- 6280149 TI - Molecular cloning of rabbit gamma heavy chain mRNA. AB - A cDNA library of rabbit spleen mRNA was screened for immunoglobulin heavy chain sequences. In this paper we report the nucleotide sequence of two cDNA clones containing part of the constant region of the rabbit gamma heavy chain mRNA. The sequence encodes part of the CH2 domain (amino acids 268 to 340), the entire CH3 domain (amino acids 341 to 447) and the 3' untranslated region. This nucleotide sequence has been compared to the corresponding sequences of mouse gamma 1, gamma 2a and gamma 2b genes. The homologies between rabbit gamma chain gene sequence and each of the mouse gamma chain gene sequences are of the same magnitude order. This comparison shows that the CH2 domains are more homologous to each other than CH3 domains or 3' untranslated sequences. The presence of species specific nucleotide positions suggests that mouse gamma chain genes could have evolved from a common ancestor shortly after the mouse-rabbit species separation. Genomic blot analysis of rabbit liver DNA with the rabbit C gamma probes shows a limited number of related sequences, with little restriction site polymorphism between individual rabbits. PMID- 6280150 TI - Benzpyrene groups bind preferentially to the DNA of active chromatin in human lung cells. AB - The cells of the bronchial epithelium of man are targets for benzo(a)pyrene carcinogenesis. When cultures of these cells, and of non-target fibroblasts, are exposed to [3H]-benzo(a)pyrene, we find that the epithelial cells metabolise and bind to DNA far greater amounts of benzpyrene than do fibroblasts. By analysis of nuclei of benzpyrene-treated cells for sensitivity to limited digestion with pancreatic DNase I, we have shown that benzpyrene groups bind initially to the DNA of expressed (DNase I sensitive) regions of chromatin in both cell types. Covalent binding of benzpyrene groups to non-expressed (DNase I resistant) regions follows rapidly in the target epithelial cells. These maintain high levels of carcinogen adducts in their DNA. In fibroblasts, benzpyrene group binding to non-expressed DNA occurs more slowly and active removal of adducts from the DNA is evident. PMID- 6280152 TI - Physical and gene mapping of chloroplast DNA from Atriplex triangularis and Cucumis sativa. AB - A rapid and simple method for constructing restriction maps of large DNAs (100 200 kb) is presented. The utility of this method is illustrated by mapping the Sal I, Sac I, and Hpa I sites of the 152 kb Atriplex triangularis chloroplast genome, and the Sal I and Pvu II sites of the 155 kb Cucumis sativa chloroplast genome. These two chloroplast DNAs are very similar in organization; both feature the near-universal chloroplast DNA inverted repeat sequence of 22-25 kb. The positions of four different genes have been localized on these chloroplast DNAs. In both genomes the 16S and 23S ribosomal RNAs are encoded by duplicate genes situated at one end of the inverted repeat, while genes for the large subunit of ribulose-1,5-bisphosphate carboxylase and a 32 kilodalton photosystem II polypeptide are separated by 55 kb of DNA within the large single copy region. The physical and genetic organization of these DNAs is compared to that of spinach chloroplast DNA. PMID- 6280151 TI - Digestion of highly modified bacteriophage DNA by restriction endonucleases. AB - The ability of thirty Type II restriction endonucleases to cleave five different types of highly modified DNA has been examined. The DNA substrates were derived from relatively large bacteriophage genomes which contain all or most of the cytosine or thymine residues substituted at the 5-position. These substituents were a proton (PBS1 DNA), a hydroxymethyl group (SP01 DNA), a methyl group (XP12 DNA), a glucosylated hydroxymethyl group (T4 DNA), or a phosphoglucuronated, glucosylated 4,5-dihydroxypentyl group (SP15 DNA). Although PBS1 DNA and SP01 DNA were digested by most of the enzymes, they were cleaved much more slowly than was normal DNA by many of them. 5-Methylcytosine-rich XP12 DNA and the multiply modified T4 and SP15 DNAs were resistant to most of these endonucleases. The only enzyme that cleaved all five of these DNAs was TaqI, which fragmented them extensively. PMID- 6280153 TI - tRNA genes are found between 16S and 23S rRNA genes in Bacillus subtilis. AB - There are at least nine, and probably ten, ribosomal RNA gene sets in the genome of Bacillus subtilis. Each gene set contains sequences complementary to 16S, 23S and 5S rRNAs. We have determined the nucleotide sequences of two DNA fragments which each contain 165 base pairs of the 16S rRNA gene, 191 base pairs of the 23S rRNA gene, and the spacer region between them. The smaller space region is 164 base pairs in length and the larger one includes an additional 180 base pairs. The extra nucleotides could be transcribed in tRNAIIe and tRNA Ala sequences. Evidence is also presented for the existence of a second spacer region which also contains tRNAIIe and tRNA Ala sequences. No other tRNAs appear to be encoded in the spacer regions between the 16S and 23S rRNA genes. Whereas the nucleotide sequences corresponding to the 16S rRNA, 23S rRNA and the spacer tRNAs are very similar to those of E. coli, the sequences between these structural genes are very different. PMID- 6280154 TI - Nature of an inserted sequence in the mitochondrial gene coding for the 15S ribosomal RNA of yeast. AB - The small ribosomal RNA, or 15S RNA, or yeast mitochondria is coded by a mitochondrial gene. In the central part of the gene, there is a guanine-cytosine (GC) rich sequence of 40 base-pairs, flanked by adenine-thymine sequences. The GC rich sequence is (5') TAGTTCCGGGGCCCGGCCACGGAGCCGAACCCGAAAGGAG (3'). We have found that this sequence is absent in the 15S rRNA gene of some strains of yeast. When present, it is transcribed into the mature 15S rRNA to produce a longer variant of the RNA. Sequences identical or closely related to this GC-rich sequence are present in many regions of the mitochondrial genome of Saccharomyces cerevisiae. The 5' and 3' terminal structures of all these sequences are highly constant. PMID- 6280155 TI - Differential stabilization by netropsin of inducible B-like conformations in deoxyribo-, ribo- and 2'-deoxy-2'-fluororibo-adenosine containing duplexes of (dA)n . (dT)n and (dA)n . (dU)na. AB - Six polynucleotide duplexes containing polydeoxyadenylic acid, polyadenylic acid or poly-2'-deoxy-2'-fluoro-adenylic acid in one strand, and polydeoxyuridylic acid or polydeoxythymidylic acid in the other strand have been studied by circular dichroism, ionic strength dependence of melting temperatures and binding of the DNA specific antibiotic netropsin. Circular dichroism spectra of (dA)n . (dT)n and (dA)n . (dU)n indicated the presence of the B-form of DNA, while those of (dAfl)n . (dT)n and (rA)n . (dT)n (and the corresponding (dU)n hybrids) indicated the presence of the A-form. (dAfl)n . (dT)n and (dAfl)n . (dU)n bound netropsin only slightly less than the (dA)n containing duplexes, while replacement by (rA)n decreased netropsin binding to a large degree. Since netropsin requires B-DNA for binding, it is concluded that the A to B transition is facilitated in the case of fluorine substitution in the sugar moiety, while the 2'-OH group greatly limits this conformational change. PMID- 6280156 TI - Expression in Escherichia coli of chemically synthesized gene for a novel opiate peptide alpha-neo-endorphin. AB - Chemically synthesized alpha-neo-endorphin gene was fused to the Escherichia coli beta-galactosidase gene on the plasmid pKO13. The resulting recombinant DNA was used to transform E. coli cells. Radioimmunoassay for alpha-neo-endorphin in CNBr treated bacterial cells showed that alpha-neo-endorphin was synthesized at approximately 5 x 10(5) molecules per single E. coli cell. One of the transformants, WA802/p alpha NE2, was used for alpha-neo-endorphin purification. From 10.9 g of wet cells, we isolated 4 mg of chemically pure and biologically active alpha-neo-endorphin. PMID- 6280157 TI - Effect of high mobility group nonhistone proteins HMG-20 (ubiquitin) and HMG-17 on histone deacetylase activity assayed in vitro. AB - We have used a method previously described by Reeves and Candido (1) to partially release histone deacetylase from cell nuclei together with putative regulators of the enzyme. Histone deacetylase released from testis cell nuclei and its putative regulators were separated by gel filtration in Sepharose 6B. A peak of low molecular weight contains a heat-stable factor that stimulate histone deacetylase in vitro. Many of the properties of the activator coincide with those of the protein HMG-20 (ubiquitin). Ubiquitin isolated from testis cell nuclei stimulated histone deacetylase in vitro. It has been suggested that HMG-17 partially inhibits histone deacetylase in Fried cell nuclei (2). In our system, HMG-17 shows no inhibitory effect on histone deacetylase activity PMID- 6280158 TI - Lead poisoning: something in the air. PMID- 6280159 TI - Unfilled, filled, and microfilled composite resins. PMID- 6280160 TI - Parasitic bowel disease: three pathogens important in primary care. AB - A high index of suspicion and careful application of diagnostic methods are essential for accurate diagnosis of parasitic bowel diseases. The varied clinical spectrum of giardiasis, amebiasis, and strongyloidiasis emphasizes the need to consider these pathogens when patients present with gastrointestinal complaints. Giardiasis should be suspected in patients, especially returned travelers, with unexplained increase in stool frequency, particularly with bloating, flatulence, or vague systemic symptoms. Amebiasis must be considered in the differential diagnosis of any patient who presents with persistent diarrhea or signs of inflammatory bowel disease. Unexplained diarrheal illnesses associated with upper abdominal symptoms and eosinophilia should raise suspicion of the presence of strongyloidiasis. These findings in a patient with a compromised immune system or in a candidate for immunosuppressive therapy should prompt a thorough investigation to rule out this parasite, since disseminated strongyloidiasis often is fatal. PMID- 6280161 TI - Specific adsorbents for affinity chromatography of benzodiazepine binding proteins. AB - The use of affinity chromatography as means of isolating/purifying proteins which have an affinity for benzodiazepine is described. Three such drugs are employed: chlorazepate, clonazepam and delorazepam. The results presented in this paper indicate that the proposed technique only works for chlorazepate and delorazepam. In fact these benzodiazepine-Sepharose derivatives are able to retain specifically proteins from human serum and rat kidney, lung, skeletal muscle and brain. PMID- 6280162 TI - In vitro formation of multimeric DNA structures mediated by purified simian virus 40 chromatin. AB - Simian virus 40 chromatin was incubated after purification by sucrose density gradient centrifugation with various circular double-stranded DNA substrates. Monomeric rings were converted in the presence of Mg2+ to structures possessing a higher degree of complexity. Dimeric catenanes, as well as multimeric linear structures and concatemers, were generated, indicating that recombination events had occurred in vitro involving covalent linkage between different DNA molecules. Furthermore, apparently fused dimeric rings were observed. Their structures suggest that they may be recombination intermediates such as those described in a prokaryotic system [Potter, H. & Dressler, D. (1976) Proc. Natl. Acad. Sci. USA 73, 3000-3004]. Recombination did not take place between heterologous DNA substrates, as exclusively homologous multimeric DNA structures were observed. PMID- 6280164 TI - Beta-endorphin: replacement of tyrosine in position 27 by tryptophan increases analgesic potency--preparation and properties of the 2-nitrophenylsulfenyl derivative. AB - An analog of human beta-endorphin with tryptophan in position 27 has been synthetized by the solid-phase method. Bioassay of the analog showed that it had almost 4 times the analgesic potency of the parent hormone but only 68% of the opiate receptor-binding activity. The peptide is the most potent analgesic among the known synthetic analogs of beta-endorphin. The 2-nitrophenylsulfenyl derivative of the analog has been prepared and shown to have a lower analgesic potency and a higher opiate receptor-binding activity than the parent compound. PMID- 6280163 TI - Isolation and partial characterization of a 5'-nucleotidase specific for orotidine-5'-monophosphate. AB - A previously unknown 5'nucleotidase (5'-ribonucleotide phosphohydrolase, EC 3.1.3.5) (5'-Nase) specific for orotidine 5'-monophosphate (OMP) hs been discovered. This enzyme orotidine 5'-monophosphate phosphohydrolase (OMPase), was isolated from mouse liver microsomes as a separate entity from the nonspecific 5' Nase. OMPase was partially purified and is shown to cleave OMP to orotidine and inorganic phosphate. The enzyme has negligible activity towards UMP, CMP, dTMP, AMP, IMP, GMP, XMP, 6-azauridine 5'-monophosphate, 1-beta-D ribofuranosylbarbituric acid 5'-monophosphate (BMF), 2'-UMP, 3'-UMP, 2'-AMP, 3' AMP, ribose 5-phosphate and beta-glycerophosphate, all of which--with the exception of the 2' or 3' monophosphates, ribose 5'-phosphate, and beta glycerophosphate--are substrates for 5'-Nase. Both enzymes are inhibited by NaF, but only OMPase is inhibited by SF reagents. OMPase is not inhibited by orotidine, orotate, BMP, concanavalin A, or tetramisole (an alkaline phosphatase inhibitor). OMPase had a Mr 53,000, Km value of 1 mM for OMP, and Vmax value of 49 nmol/min . mg of protein at the present stage of purification. OMPase activity has also been detected in various mammalian tissues including normal human tissues, human tumor xenografts, lymphocytes, and rat liver. OMPase may be responsible, in part, for the low levels of intracellular "free" OMP and for orotidine accumulation in cells treated with 6-azauridine and patients suffering from aortic aciduria. PMID- 6280165 TI - On the enzymic mechanism of oxidative phosphorylation. AB - Oxidative phosphorylation, like substrate-level phosphorylation, involves oxidative conversion of inorganic phosphate to a reactive species followed by interaction of this species with enzyme-bound ADP to form enzyme-bound ATP. The reactive species in a phosphoryl ester in substrate-level phosphorylation and phosphonium ion of orthophosphate in oxidative phosphorylation. The coupled synthesis is mediated by a combination of two classical enzymes in substrate level phosphorylation and by a set of energy-coupled enzymes in oxidative phosphorylation. The full range of experimental evidence supporting this proposed enzymic mechanism of oxidative phosphorylation is presented as well as the rationalization of phenomena that hitherto have eluded explanation. PMID- 6280166 TI - Mossbauer studies of beef heart aconitase: evidence for facile interconversions of iron-sulfur clusters. AB - Beef heart aconitase, isolated under aerobic conditions, has been studied with Mossbauer and EPR spectroscopy. In the oxidized state, the enzyme exhibits an EPR signal at g = 2.01. The Mossbauer data show that this signal is associated with a 3Fe cluster. In dithionite-reduced aconitase, the 3Fe cluster, probably of the [3Fe-3S] type, is in a paramagnetic state of interger electronic spin (S = 2); the Mossbauer spectra exhibit al the unique features reported for proteins with 3Fe clusters. On activation of aconitase with ferrous ion, the paramagnetic 3Fe cluster of dithionite-reduced enzyme is converted into a diamagnetic (S = 0) form. Activation studies with iron enriched in either 27 Fe or 56 Fe suggest that activation transforms the 3Fe cluster into a center that has a [4Fe-4S] core. This conclusion is supported by the observation that EPR signals characteristic of reduced [4Fe-4S] clusters can be elicited under appropriate conditions. It has frequently been assumed that the activation of aconitase with Fe2+ produces an active site containing a single ferrous ion. The data reported here suggest that a ferrous ion is used to rebuild a [4Fe-4S] cluster. PMID- 6280168 TI - Long-lived cytoplasmic factors that suppress adrenal steroidogenesis. AB - Adrenal cells secrete steroids after stimulation with corticotropin (ACTH), whereas cells reconstructed by fusing adrenal cell nuclei to fibroblast cytoplasms are temporarily (2-3 wk) unresponsive to ACTH. In this report, we characterize this inhibition by using "cybrids" (cytoplasmic hybrids) isolated by either genetic selection or a new procedure that utilizes the fluorescence activated cell sorter and the vital mitochondrial dye rhodamine 123. Such cybrids, which contain both adrenal and fibroblast cytoplasmic components, are unable to produce steroids, suggesting the existence of cytoplasmic inhibitory factors. In order to elucidate this cytoplasmic inhibition of steroidogenesis, techniques are described that test the contribution of fibroblast mitochondria to this phenomenon. The first technique utilizes purified mitochondria, isolated from chloramphenicol (CAP)-resistant fibroblasts, to confer CAP resistance on an otherwise sensitive adrenal cell. The resulting CAPr cells, termed mitochondrial transformants, are responsive to ACTH. The second technique utilizes a procedure for isolating small fragments of cytoplasm (microcytospheres) from fibroblasts. Microcytospheres, which do not contain mitochondria, are stained with rhodamine 18, a vital membrane dye, and then fused to unstained adrenal cells. The fusion products are then isolated with the fluorescence-activated cell sorter. Approximately 30% of the fusion products are inhibited in their ability to respond to ACTH. These results suggest that the fibroblast cytoplasm contains nonmitochondrial long-lived inhibitory factors that temporarily suppress steroidogenic function in adrenal cells. PMID- 6280167 TI - Effect of methylation on expression of microinjected genes. AB - The cloned genes for the simian virus 40 large tumor antigen and for herpes simplex virus (HSV) thymidine kinase (TK) were methylated with EcoRI methylase. The genes were microinjected into the nuclei of TK-deficient (tk-) cells, and expression of the genes was determined by immunofluorescence staining for the simian virus 40 large tumor antigen and by [3H]thymidine incorporation followed by autoradiography for HSV TK. We found that methylation of the simian virus 40 gene, under EcoRI or EcoRI* conditions, resulting in methylation at sites within the gene and in the surrounding sequences, has no effect on expression of the large tumor antigen when the gene is manually microinjected into mammalian nuclei. However, methylation of the HSV tk gene at the two EcoRI sites markedly reduces or abolishes the expression of this gene. One of the EcoRI sites of HSV tk is approximately 1.1 kilobases downstream from the 3' end of the gene and is believed to have no regulatory function in the expression of the tk gene. The other EcoRI site is 79 base pairs upstream from the 5' end of the gene and has considerable homology to the regulatory sequence proposed by [Benoist C., O'Hare, K., Breathnach, R., & Chambon, P. (1980) Nucleic Acids Res. 8, 127-142]. Our results are direct proof that methylation can alter gene expression and also that the effect depends strictly on the sites that are methylated. PMID- 6280169 TI - Epidermal growth factor and insulin cause normal chicken heart mesenchymal cells to proliferate like their Rous sarcoma virus-infected counterparts. AB - Normal chicken heart mesenchymal cells at low culture density are proliferatively quiescent in a physiological culture medium containing heparinized, heat inactivated, chicken plasma at 10%. Rous sarcoma virus-infected chicken heart mesenchymal cells, on the other hand, proliferate maximally in this same medium, undergoing a 60-fold increase in cell number during 4 days of exponential growth. When normal heart mesenchymal cells are cultured for 4 days in the presence of epidermal growth factor at 1 micrograms/ml they undergo a 16-fold increase in number, with graded responses to lower concentrations of the factor. In the presence of insulin at 10 micrograms/ml, normal heart mesenchymal cells multiply by a factor of 3 over a 4-day period. The addition of epidermal growth factor (1 microgram/ml) and insulin (10 micrograms/ml) to cultures of normal chicken heart mesenchymal cells causes these cells to proliferate at a rate comparable to that of their RSV-infected counterparts. PMID- 6280170 TI - ESR studies of O2 uptake by Chinese hamster ovary cells during the cell cycle. AB - Magnetic interactions between dissolved oxygen and nitroxide radical spin probes lead to broadening of the ESR lines. We have used a closed-chamber method based on this property to determine the maximum rate of O2 uptake per cell (Vmax per cell) in cultured mammalian cells. A suitable spin probe and a cell suspension are mixed in an aerated medium, and the rate of disappearance of dissolved O2 is measured. The effects of temperature, pH, and microwave power on the determination of dissolved oxygen in solution were studied. For asynchronous Chinese hamster ovary cells, oxygen uptake is 3.8 X 10(7) oxygen molecules per cell per sec and appears to be enzymatically limited at oxygen concentrations greater than 10 microM. About 5-10 X 10(5) cells were used for each measurement, making it possible to study mitotically synchronized cells. Using this method, we have found that Vmax per unit cell volume changes during the cell cycle of Chinese hamster ovary cells from a minimum in mitosis to maxima in both G1 and late S phases. Advantages and limitations of spin probes for studying the O2 uptake of intact cells are discussed. PMID- 6280171 TI - Monoclonal antibody to transferrin receptor blocks transferrin binding and inhibits human tumor cell growth in vitro. AB - A murine hybridoma has been obtained that produces a monoclonal antibody against the human transferrin receptor. In contrast to previously characterized monoclonal antibodies that recognize the transferrin receptor, this antibody, designated 42/6, blocks the binding of transferrin to its receptor and inhibits the growth of the human T leukemic cell line, CCRF-CEM, in vitro. Inhibition of cell growth was dose dependent, and as little as 2.5 micrograms of purified antibody per ml had a detectable effect, even though transferrin was present in the tissue culture medium in large molar excess. Cells grown in the presence of antibody for 7 days accumulated in S phase of the cell cycle. The addition of iron to antibody-treated cultures in the form of ferric complexes or ferrous sulfate did not overcome the growth inhibitory effects of the anti-transferrin receptor antibodies. This result suggests that either transferrin is the only means by which CCRF-CEM leukemic cells can be provided with sufficient iron in vitro or that other factors in addition to iron starvation are involved in the antibody-mediated growth inhibition. The inhibition of cell growth by 42/6 monoclonal antibody suggests that monoclonal antibodies against proliferation associated cell surface antigens, such as the transferrin receptor, may be useful pharmacological reagents to modify cell growth in vitro. PMID- 6280172 TI - Murine erythroleukemia cell differentiation: DNase I hypersensitivity and DNA methylation near the globin genes. AB - The sensitivity to digestion by DNase I of chromatin containing the alpha- and beta(major)-globin genes and the pattern of DNA methylation near these genes was examined during hexamethylenebisacetamide (HMBA)-mediated erythroid differentiation of murine erythroleukemia cells (MELC). In uninduced and induced cells, the chromatin regions containing the alpha- and beta-(major)-globin genes are more sensitive to digestion by DNase I than is the region containing an immunoglobulin gene (Igalpha) not expressed during erythroid differentiation. However, at low concentrations of DNase I, a 6- to 10-fold increase in site specific cleavages was generated in chromatin regions near both the alpha- and beta(major)-globin genes in cells induced to differentiate by HMBA. The DNase I hypersensitive site near the beta(major)-globin gene maps to a small region near the 5' terminus of the gene. No detectable change in the pattern of DNA methylation around either the alpha- or beta-globin genes was observed during HMBA-mediated erythroid differentiation. Of the potentially methylated sites assayed and mapped near the beta(major)-globin gene, one site is fully methylated, one is partially methylated, and one is unmethylated both in uninduced and induced cells. Many (but not all) sites assayed near the alpha globin genes are unmethylated in both uninduced and induced cells. These results show that specific alterations of chromatin structure occur during MELC differentiation and suggest that these changes may not involve alterations in the pattern of DNA methylation. PMID- 6280173 TI - Mapping of the vaccinia virus thymidine kinase gene by marker rescue and by cell free translation of selected mRNA. AB - A selective plaque assay that uses thymidine kinase (TK)-deficient human 143 cells was developed to titer mixtures of TK(+) and TK(-) vaccinia virus. With this assay it could be shown that methotrexate-resistant TK(+) virus was formed in cells coinfected with TK(-) virus and wild-type virus DNA. By substituting vaccinia DNA fragments cloned in plasmids for virion DNA, this marker rescue system provided the basis for mapping the TK gene. Of the 15 HindIII fragments, only J could rescue five independently derived TK(-) mutants. This 5000-base-pair (bp) fragment maps approximately 80,000 bp from the left-end of the 180,000-bp vaccinia genome. Marker rescue could be detected with 18 ng or less of plasmid and was proportionate to DNA concentration. The resistance to methotrexate of the TK(+) recombinants was shown to be due to TK synthesis. Evidence that the HindIII J fragment contains the structural TK gene and not a regulatory element was demonstrated by the synthesis of active TK in a cell-free system programmed with mRNA selected by hybridization to the plasmid. Previous studies [Belle-Isle, H., Venkatesan, S. & Moss, B. (1981) Virology 112, 306-317] indicated that mRNAs coding for three immediate early polypeptides with molecular weights of 41,000, 21,000, and 17,000 map within HindIII J. The mapping of the easily selectable vaccinia virus TK gene now opens the way to genetic manipulations that should increase our understanding of vaccinia virus gene expression and facilitate the use of vaccinia virus as an efficient cloning vector for foreign genes. PMID- 6280174 TI - At least two regions of the viral genome determine the oncogenic potential of avian leukosis viruses. AB - Recombinants of oncogenic and nononcogenic avian leukosis viruses were tested for their oncogenic potential in chickens. The results indicate that at least two regions of the viral genome determine the oncogenic potential of these viruses. The first region contains sequences that control viral mRNA synthesis. These sequences determine the potential of a virus to induce a low incidence of lymphomas, carcinomas, chondrosarcomas, fibrosarcomas, and osteopetrosis. The second region lies outside the sequences that control viral mRNAs synthesis. These sequences determine the ability of a virus to induce a high incidence of lymphomas or osteopetrosis. PMID- 6280175 TI - Primary structure analysis of the major internal protein p24 of human type C T cell leukemia virus. AB - A human type C retrovirus [human T-cell leukemia (lymphoma) virus; HTLV], recently isolated from young adult patients with cutaneous T-cell lymphoma or leukemia, was not detectably related to other known animal retroviruses in molecular hybridization studies, by comparison of reverse transcriptase and the major core protein p24. The p24 core protein was purified to homogeneity. The amino acid composition, the COOH-terminal amino acid, and the NH(2)-terminal amino acid sequence of the first 25 residues of this major internal structural protein were determined. These results were then compared to the known structure of the internal core protein of other retroviruses. The compositional data reveal that HTLV p24 is chemically distinct from p30-p24 of other animal retroviruses, in agreement with the earlier immunological analyses. However, HTLV p24 shares the common NH(2)-terminal proline and COOH-terminal leucine of all mammalian type C viral p30s. In addition, like bovine leukemia virus (BLV), HTLV lacks the common prolylleucylarginine tripeptide and the larger conserved region found near the NH(2) terminus of the other mammalian type C viral p30s. Alignment of the amino acid sequence of HTLV p24 with previously determined sequences of other retrovirus proteins, including BLV p24, reveals statistically significant sequence homology only to BLV. The results reported here demonstrate that HTLV p24 is related to but chemically distinct from the major core protein of other retroviruses. Similarly, previous results showed that there was no immunological crossreactivity of the p24 protein and reverse transcriptase of HTLV with other retroviruses, including BLV, and no nucleic acid sequence homology. However, the present results, combined with the common size of the p24 and reverse transcriptase, suggest that HTLV may be closer to BLV than any other known retrovirus. PMID- 6280177 TI - Purification and properties of the uvrA protein from Escherichia coli. AB - The uvrA+ gene product from Escherichia coli was purified to apparent homogeneity; the assay measured its ability to restore repair endonuclease activity in extracts from uvrA mutated cells. The uvrA protein is a 115,000 molecular weight DNA-binding protein having higher affinity for single-stranded than double-stranded DNA. It does not introduce single-strand breaks or alkali labile bonds in native or UV-irradiated DNA, but it catalyzes hydrolysis of ATP to ADP and Pi. The ATPase activity is not DNA dependent and has a Km of 0.23 mM, which corresponds to the Km for the ATP requirement of the UV-endonuclease reaction catalyzed by the combined uvrA+, uvrB+, and uvrC+ gene products. ADP and adenosine 5'-[gamma-thio]triphosphate both inhibit the uvrA ATPase as well as the uvrABC endonuclease and also prevent specific binding of the uvrA proteins to UV irradiated DNA. These results indicate that both the DNA-binding property and the ATPase activity of the uvrA protein are essential for uvrABC endonuclease activity and that the ATP requirement of the endonuclease reaction is determined by uvrA ATPase. PMID- 6280176 TI - Analysis of the sequence of amino acids surrounding sites of tyrosine phosphorylation. AB - We have identified the single phosphorylated tyrosine in p60src, the transforming protein of Rous sarcoma virus, as part of the sequence. NH2-Arg-Leu-Ile-Glu-Asp Asn-Glu-Tyr(P)-Thr-Ala-Arg-COOH. Therefore, this is a sequence that is recognized efficiently by a tyrosine protein kinase in vivo. Phosphorylation of tyrosine in cellular proteins appears to play a role in malignant transformation by four classes of genetically distinct RNA tumor viruses. Phosphorylated tyrosines in several other proteins resemble of the tyrosine in p60src in that they are located 7 residues to the COOH-terminal side of a basic amino acid and either 4 residues to the COOH-terminal side of, or in close proximity to, a glutamic acid residue. Therefore it is possible that these features play a role in the selection of sites of phosphorylation by some tyrosine protein kinases. However, several clear exceptions to this rule exist. PMID- 6280178 TI - Molecular cloning and physical mapping of varicella-zoster virus DNA. AB - Varicella-zoster virus (VZV) DNA was cleaved with restriction endonuclease EcoRI, and most of the resulting fragments were successfully cloned in the phage vector lambda gtWES . lambda B. Double digestions of cloned fragments with EcoRI and BamHI and hybridizations to blot-transferred BamHI digests of VZV DNA were used to construct a physical map of the genome. The molecular termini of the DNA were identified by restriction enzyme analysis after exonuclease III digestion. The data indicate that VZV DNA exists in two isomeric forms that differ by inversion of one short terminal genome segment. Electron microscopic studies revealed that the short genome segment consists of a terminal revealed that the short genome segment consists of a terminal sequence of about 3.4 X 10(6) daltons that is separated from an internal inverted repeat of itself by a 5.8 X 10(60)-dalton unique DNA segment. PMID- 6280179 TI - Clonidine p-isothiocyanate, an affinity label for alpha 2-adrenergic receptors on human platelets. AB - Exposure of intact human platelets or platelet membranes to the clonidine analog clonidine p-isothiocyanate (clonidine-NCS), followed by extensive washing, results in the loss of [3H]yohimbine binding to platelet alpha 2-receptors. In addition, exposure of intact platelets to clonidine-NCS, followed by extensive washing, results in the loss of of epinephrine-induced inhibition of adenylate cyclase activity [ATP pyrophosphate-lyase (cyclizing), EC 4.6.1.1] in frozen- thawed platelets and in purified platelet membranes. This effect is dependent on time and concentration (t 1/2 at 30 degrees C is less than 15 min; half-maximal effect occurs with clonidine-NCS at less than 10 microM). Clonidine-NCS appears to interact by irreversibly blocking the platelet alpha 2-receptors because (i) it abolishes alpha 2-receptor effects of adenylate cyclase activity (i.e., epinephrine-induced inhibition of basal and prostaglandin E1-stimulated activity) while not altering other cyclase activity (basal, prostaglandin E1-stimulated, and NaF-stimulated) and (ii) its effect on both [3H]yohimbine binding and epinephrine-induced inhibition of adenylate cyclase can be specifically prevented by alpha-agonists [(-)-epinephrine and clonidine] and alpha-antagonists (yohimbine and phentolamine). These observations indicate that clonidine-NCS is an effective affinity label for platelet alpha 2-receptors. PMID- 6280180 TI - Molecular cloning of murine endogenous viral sequences and expression of a newly constructed recombinant murine leukemia virus DNA in transfected mink cells. AB - In the process of molecularly cloning unintegrated proviral DNA from NIH-3T3 mouse cells infected with Rauscher murine leukemia virus, we observed the occurrence of clones with inserts smaller than the expected Rauscher murine leukemia virus fragments. The insert of one of these clones, lambda.Xe-1, was characterized in more detail. It had a size of 3.5 kilobases. The restriction map was similar but not identical to that of the envelope regions of Moloney and Rauscher murine leukemia viruses. After ligation to previously cloned Moloney murine leukemia viral sequences and transfection of the ligated DNA into mink lung cells a nondefective xenotropic murine leukemia virus, XH-19, was isolated. Restriction mapping of proviral DNA isolated from mink lungs cells chronically infected with XH-19 showed the presence of Moloney murine leukemia virus-derived sequences coupled to xenotropic viral sequences. PMID- 6280181 TI - Site-specific cleavage/packaging of herpes simplex virus DNA and the selective maturation of nucleocapsids containing full-length viral DNA. AB - Defective genomes present in serially passaged herpes simplex virus (HSV) stocks have been shown to consist of tandemly arranged repeat units containing limited sets of the standard virus DNA sequences. Invariably, the HSV defective genomes terminate with the right (S component) terminus of HSV DNA. Because the oligomeric forms can arise from a single repeat unit, it has been concluded that the defective genomes arise by a rolling circle mechanism of replication. We now report on our studies of defective genomes packaged in viral capsids accumulating in the nuclei and in mature virions (enveloped capsids) translocated into the cytoplasm of cells infected with serially passaged virus. These studies have revealed that, upon electrophoresis in agarose gels, the defective genomes prepared from cytoplasmic virions comigrated with nondefective standard virus DNA (M(r) 100 x 10(6)). In contrast, DNA prepared from capsids accumulating in nuclei consisted of both full-length defective virus DNA molecules and smaller DNA molecules of discrete sizes, ranging in M(r) from 5.5 to 100 x 10(6). These smaller DNA species were shown to consist of different integral numbers (from 1 to approximately 18) of defective genome repeat units and to terminate with sequences corresponding to the right terminal sequences of HSV DNA. We conclude on the basis of these studies that (i) sequences from the right end of standard virus DNA contain a recognition signal for the cleavage and packaging of concatemeric viral DNA, (ii) the sequence-specific cleavage is either a prerequisite for or occurs during the entry of viral DNA into capsid structures, and (iii) DNA molecules significantly shorter than full-length standard viral DNA can become encapsidated within nuclear capsids provided they contain the cleavage/packaging signal. However, capsids containing DNA molecules significantly shorter than standard virus DNA are not translocated into the cytoplasm. PMID- 6280183 TI - Membrane potential changes induced by the ouabain-like compound extracted from mammalian brain. AB - The electrical membrane potential (delta psi) of chicken embryo fibroblasts in tissue culture was determined to be -30.5 +/- 2.9 mV as measured by distribution of the lipophilic [3H]tetraphenylphosphonium cation (Ph4P+). Stimulation of the electrogenic activity of the Na+,K+-ATPase by the ionophore monensin induces a hyperpolarization of approximately 47 mV and a new delta psi of -77.3 +/- 5.7 mV. The effects of the cardiac glycoside ouabain and an "ouabain-like compound" (OLC), which was extracted and partially purified from sheep brain, were contrasted using both the resting and hyperpolarized fibroblasts. Addition of OLC or ouabain to the incubation medium for short periods of time does not alter the cells' resting delta psi. However, OLC and ouabain block monensin-induced hyperpolarization. The inhibitory effects of OLC, like ouabain, are dose dependent, with half-maximal inhibition occurring at an amount of OLC equivalent to that found in 1.6 g of brain (wet weight) per ml and at 0.85 microM ouabain. In addition, the maximal actions of ouabain and OLC are not additive. These results show that the endogenous OLC specifically affects the delta psi of intact cells by a mechanism analogous to that of ouabain--i.e., inhibition of the Na+,K+ ATPase. PMID- 6280182 TI - Angiotensin increases Na+ entry and Na+/K+ pump activity in cultures of smooth muscle from rat aorta. AB - Angiotensin markedly altered the Na+ permeability of smooth muscle cells cultured from explants of rat aorta. The rate of net Na+ uptake was followed in the presence of ouabain in order to block Na+ efflux via the Na+/K+ pump. Angiotensin II (AII) or angiotensin III (AIII) increased net Na+ uptake by about 3-fold. Maximal stimulation of Na+ uptake was produced by about 10 nM AII. Bradykinin and the angiotensin antagonist [Sar1, Ileu5, Ala8]AII had no significant effect on net Na+ uptake. Angiotensin also enhanced the activity of the Na+/K+ pump, which was assayed by following the rate of ouabain-sensitive 86Rb+ uptake by the cells. AII and AIII nearly doubled ouabain-sensitive 86Rb+ uptake, but bradykinin, norepinephrine, and [Sar1, Ileu5, Ala8]AII had no effect. In the presence of ouabain, 86Rb+ uptake was not significantly affected by AII or AIII, indicating that angiotensin did not alter passive permeability to Rb+. Loading the cells with Na+, either by incubation in K+-free medium or exposure to the Na+-selective ionophore monensin, markedly increased ouabain-sensitive 86RB+ uptake. This result indicates that the activity of the Na+/K+ pump is limited by the low level of Na+ that is normally in the cells. AII had no effect on the activity of the Na+/K+ pump in Na+-loaded cells. These results suggest that AII or AIII stimulates the Na+/K+ pump in cultured aortic muscle cells by increasing its Na+ supply. PMID- 6280184 TI - RNA ligation via 2'-phosphomonoester, 3'5'-phosphodiester linkage: requirement of 2',3'-cyclic phosphate termini and involvement of a 5'-hydroxyl polynucleotide kinase. AB - Extracts of wheat germ contain a RNA ligase activity that catalyzes the conversion of linear polyribonucleotides into covalently closed circles. As reported previously, this enzyme joins two ends of a RNA substrate via a 2' phosphomonoester, 3',5'-phosphodiester linkage. In the present work we provide evidence that a 2',3'-cyclic phosphate group at the 3' terminus is required for RNA ligation and that the 5'-hydroxyl end is phosphorylated before the two RNA ends are joined. We report on the presence of 5'-hydroxyl polynucleotide kinase and polynucleotide 2',3'-cyclic phosphate 3'-phosphodiesterase activities in wheat germ extracts. A possible involvement of these enzymes in the ligation process and a potential role of the newly described ligation pathway in RNA processing are discussed. PMID- 6280185 TI - Polyoma mutants that productively infect F9 embryonal carcinoma cells do not rescue wild-type polyoma in F9 cells. AB - Mouse embryonal carcinoma cells are refractory to infection by wild-type polyoma virus, the infection process apparently being blocked at a stage after adsorption and penetration but before early protein synthesis. Polyoma virus mutants capable of productive infection of mouse embryonal carcinoma cells have been isolated and these mutants all have DNA sequence alterations in a noncoding region near the origin of replication of the viral genome. PyF101 and PyF441 are two mutants selected for their ability to infect the embryonal carcinoma cell line F9. Here we show that these PyF mutants do not rescue replication of wild-type polyoma during a mixed infection of F9 cells. The mutant and wild-type DNAs were distinguished on the basis of restriction fragments obtained by digestion with Msp I or BstNI, and no wild-type DNA was detected in F9 cells coinfected with wild-type polyoma and with either PyF101 or PyF441. The mutant viruses do not appear to inhibit wild-type replication during a mixed infection because both mutant and wild-type DNAs can replicate efficiently in coinfected 3T6 cells which are permissive for both mutant and wild-type viruses. A double mutant having the PyF101 mutation and the ts-25E temperature-sensitive mutation in polyoma large tumor antigen was constructed and found to be temperature-sensitive for replication in F9 cells. This double mutant, designated PyFts-1, can be rescued in F9 cells at the restrictive temperature by coinfection with PyF441. These results suggest that the PyF mutations affect two processes in F9 cells, one involving expression of polyoma early genes and a second involving viral DNA replication. PMID- 6280186 TI - A monkey Alu sequence is flanked by 13-base pair direct repeats by an interrupted alpha-satellite DNA sequence. AB - A member of the Alu family, the dominant family of short interspersed repeated DNA sequences in primates, interrupts a cloned repeat unit of African green monkey alpha-satellite DNA. The Alu is immediately flanked by 13-base-pair duplications of the known sequence of the satellite at the site of insertion. These observations support the idea that Ala family members may be moveable elements. PMID- 6280188 TI - Thyrotropin stimulation of the ADP-ribosyltransferase activity of bovine thyroid membranes. AB - Thyrotropin increases the ADP-ribosylation activity of bovine thyroid membranes. Rapid ADP-ribosylation of membrane components is followed by increasing ADP ribosylation of components in the supernatant of the reaction mixture. One of the major membrane proteins ADP-ribosylated in the thyrotropin-stimulated reaction has an approximate molecular weight of 40,000; this same protein is also a major ADP-ribosylated product of the A promoter of cholera toxin and appears to be related to the G regulatory subunit of the adenylate cyclase complex. The ADP ribosylated products appearing in the supernatant solution comigrate with thyrotropin and preparations of 125I-labeled alpha subunit of thyrotropin; the alpha subunit, but not the beta subunit, of thyrotropin can be ADP-ribosylated by the membrane ADP-ribosyltransferase activity. NAD can be shown to enhance the ability of thyrotropin to stimulate the adenylate cyclase activity of bovine thyroid membrane preparations and of membrane preparations of a rat thyroid tumor whose adenylate cyclase activity is otherwise unresponsive to thyrotropin. The beta subunit of thyrotropin inhibits thyrotropin stimulation of both the ADP ribosylation and adenylate cyclase activities of the thyroid membrane. PMID- 6280187 TI - H-2Kk and vesicular stomatitis virus G proteins are not extensively associated in reconstituted membranes recognized by T cells. AB - It is shown that liposomes containing (i) a fluorescein-labeled murine histocompatibility antigen (FITC-H-2Kk) and the G protein of vesicular stomatitis virus or (ii) H-2Kk and fluorescein-labeled viral protein (FITC-G) can elicit H-2 restricted syngeneic antiviral cytotoxic T cells as assayed by 51Cr release from appropriate virus-infected target cells. Fluorescence recovery after photobleaching was used to measure the diffusion coefficients of these reconstituted proteins in four different samples: (i) FITC-H-2Kk; (ii) FITC-H-2Kk and G; (iii) FITC-G; and (iv) FITC-G and H-2Kk. The same rate of lateral diffusion (D = 1 x 10(-8) cm2/sec at 37 degrees C in 25% cholesterol/75% dimyristoylphosphatidylcholine) was obtained in every case. Both proteins, fluorescent as well as nonfluorescent, could be patched by using specific antibodies. When G was patched with antibody, FITC-H-2Kk did not copatch. When H 2Kk was patched with antibody FITC-G did not copatch. These diffusion and patching measurements rule out the possibility that these proteins have either extensive oligomeric associations or strong specific pairwise associations. PMID- 6280189 TI - Isopeptidase: a novel eukaryotic enzyme that cleaves isopeptide bonds. AB - In an attempt to clarify the regulatory mechanism that accounts for the shift of protein A24 in the mitotic cycle, we demonstrated the existence of an enzyme, provisionally termed isopeptidase, that cleaves A24 stoichiometrically into histone H2A and ubiquitin. Properties of this enzyme are (i) most eukaryotes, including mammals, amphibia, chicken, and yeast, contain isopeptidase in the cytoplasm; (ii) a significant increase in enzyme binding to chromatin occurs when cells enter mitosis; (iii) Escherichia coli does not contain isopeptidase; (iv) isopeptidase has a molecular weight of 38,000; (v) at an ionic strength that induces globular conformation of H2A, isopeptidase activity is repressed; (vi) a SH group is an essential cofactor; and (vii) most divalent cations (except Mg2+ and Ca2+) are inhibitory. In view of the stoichiometric conversion of A24 into H2A and ubiquitin by isopeptidase in vitro, A24 probably contains a Gly-Gly dipeptide in isopeptide linkage but no other intervening polypeptides. Since ubiquitin in various eukaryotes binds to protein other than H2A, and is proteolytically released, isopeptidase probably acts on isopeptide bonds in general and not uniquely on those of A24. Inasmuch as isopeptidase is present throughout the cell cycle, the level of A24 in chromatin appears to be controlled by a balance between isopeptidase and an as yet unestablished H2A-ubiquitin ligase. PMID- 6280190 TI - Proteins antigenically related to the human erythrocyte glucose transporter in normal and Rous sarcoma virus-transformed chicken embryo fibroblasts. AB - Antibody raised against the purified human erythrocyte glucose transporter specifically precipitated four proteins from normal and Rous sarcoma virus transformed chicken embryo cells: a major protein of Mr 41,000 and minor proteins of Mr 68,000, 73,000, and 82,000. The Mr 41,000 and 82,000 proteins were found only in a membrane fraction, not in the soluble fraction, and displayed a heterogeneous mobility on NaDodSO4/polyacrylamide gel electrophoresis, suggesting glycosylation. The Mr 41,000 and 82,000 proteins were increased in amount after malignant transformation in direct proportion to the increase in hexose transport rate, and the increase was dependent on the expression of the src gene product, as revealed with a temperature-conditional src mutant. We suggest that the Mr 41,000 and 82,000 proteins are the glucose transporter of chicken embryo fibroblasts, or a component of the glucose transporter. These experiments provide direct evidence that malignant transformation increases the rate of glucose transport by increasing the number of transporters in the membrane. PMID- 6280191 TI - Retrovirus long terminal repeats activate expression of coding sequences for the herpes simplex virus thymidine kinase gene. AB - In this study, we demonstrate that the long terminal repeats (LTRs) of a murine retrovirus can activate expression of heterologous gene coding sequences from which a functional promoter region has been deleted. Recombinant plasmid clones were obtained that contained both cloned fragments of Friend spleen locus-forming virus (SFFV) DNA and the herpes simplex virus (HSV) thymidine kinase (TK; ATP:thymidine 5'-phosphotransferase, EC 2.7.1.21) gene (tk). The effects of the LTR on tk expression were determined by constructing clones containing tk coding sequences with or without 5' sequences necessary for the initiation of transcription, inserted either 200 or 1200 base pairs downstream from the SFFV 5' LTR. The expression of the HSV TK protein by these clones was tested by gene transfer of the cloned into TK- mouse cells and assay of TK enzyme activity in TK+ transformants. These experiments demonstrate that: (i) the SFFV 5' LTR activates expression of tk coding sequences when these sequences are inserted 200 base pairs downstream from, and in the same orientation as, the LTR; (ii) tk is not activated when placed 1200 base pairs downstream from, and in the same orientation as, the LTR or when tk is inserted in either site in the opposite orientation as the LTR; (iii) the SFFV 5' LTR does not interfere with in vivo expression of tk when it is flanked by homologous 5' promoter sequences. The implication of these observations for retrovirus oncogenesis and animal cell genetics is discussed. PMID- 6280192 TI - Suppressor of yeast mitochondrial ochre mutations that maps in or near the 15S ribosomal RNA gene of mtDNA. AB - A polypeptide chain-terminating mutation in the yeast mitochondrial oxi 1 gene has been shown to be an ochre (TAA) mutation by DNA sequence analysis. Mitochondrially inherited revertants of this mutation include two types: In the first, the ochre codon has been changed to a sense codon by further mutation in the oxi 1 gene while, in the second, the ochre codon is still present, indicating the occurrence of an extrageneic ochre suppressor mutation. This mitochondrial ochre suppressor, termed MSU1, has been "cloned" in rho- strains of yeast and tested against other oxi 1 mutations. Several additional mutations are also suppressible, and those examined so far are also ochre mutations. MSU1 does not suppress known frameshift or missense mutations at oxi 1. Isoelectric focusing of the gene product (cytochrome oxidase subunit II) from a suppressed-mutant strain indicates that suppression does not involve insertion of charged amino acids. Physical mapping of the mtDNA retained in the MSU1-carrying rho- clones localizes the suppressor mutation to the gene coding the 15S rRNA or a site not more than 300 base pairs from it. No known tRNA genes occur this close to the 15S rRNA gene, and mtDNA from a suppressor-carrying rho- does not hybridize detectably to mitochondrial tRNAs. These results suggest that MSU1 may be an alteration in the 15S rRNA. PMID- 6280194 TI - Human papillomavirus DNA in cutaneous primary and metastasized squamous cell carcinomas from patients with epidermodysplasia verruciformis. AB - DNA extracted from squamous cell carcinomas from patients with the chronic wart disease syndrome, epidermodysplasia verruciformis, was analyzed for the presence of human papillomavirus (HPV)-specific DNA sequences by Southern blot hybridization analysis. Employing an HPV probe obtained by molecular cloning of viral DNA purified from benign warts from these patients, we have unequivocally identified HPV-specific nucleotide sequences in squamous cell carcinomas from these patients. Restriction endonuclease mapping indicated that the DNA present in the carcinomas was of the same type (type 5) as that found in the benign tumors from these patients and was present as unintegrated, free viral DNA. Moreover, we have demonstrated the presence of HPV-5 DNA in a subcutaneous metastatic tumor from one of these patients. This latter observation essentially eliminates the possibility that the HPV-5 DNA present in the malignant tumors in these patients resulted from cross-contamination from an adjacent benign warty lesion. In addition to wild-type HPV-5 DNA, both the primary and metastatic carcinomas analyzed also contained an HPV-5 DNA species lacking approximately 20% of the HPV-5 DNA genome. These subgenomic forms of HPV-5 DNA could not be detected in benign papillomas from these patients. PMID- 6280193 TI - Reversible repression and activation of measles virus infection in neural cells. AB - Conversion of acute measles virus infection to an indolent state has been achieved by treatment of infected cells of neural origin with agents that affect cyclic nucleotide metabolism. Striking results were obtained with papaverine, an inhibitor of cAMP phosphodiesterase that is capable of enhancing neural differentiation. In papaverine-treated cultures, decreased production of infectious virus was accompanied by selective disappearance of intracellular matrix proton, as detected by immunofluorescence. Viral nucleocapsid protein was enhanced in the cytoplasm while three other structural proteins--polymerase, hemagglutinin, and fusion protein--showed little change in distribution or intensity of staining. These results were specific for cells of neural origin and not observed in CV-1 or Vero cultures. cAMP, dibutyryl cAMP, 8-bromo-cAMP, and isobutylmethylxanthine all inhibited replication of virus but less so than did papaverine. Inhibition of virus replication by any of these agents was rapidly reversible, either by removal of the agent or by addition of cGMP to the culture medium and was accompanied by reappearance of the matrix protein. These results suggest that measles virus replication in neural cells depends on host factors, particularly those affecting endogenous cAMP and cGMP. Viral persistence may thus be related to the state of neural differentiation. This model system may yield information on mechanisms of recrudescence observed in some chronic diseases of the nervous system. PMID- 6280195 TI - Isolation and characterization of a monoclonal antibody against the saxitoxin binding component from the electric organ of the eel Electrophorus electricus. AB - A monoclonal hybridoma cell line secreting antibody against the saxitoxin-binding component from the eel Electrophorus electricus has been isolated. The specificity of this monoclonal antibody was established by (i) its ability to immunoprecipitate bound [3H]saxitoxin from a detergent extract of electroplax membranes in a dose-dependent manner, (ii) the inability of unrelated monoclonal antibodies to immunoprecipitate the toxin-binding activity in a similar assay, and (iii) the ability of excess unlabeled tetrodotoxin to displace [3H]saxitoxin from the immunoprecipitated component. The antibody is of the subclass IgG1 and binds specifically to a polypeptide component of Mr approximately 250,000 on NaDodSO4/polyacrylamide gels. The antigenic determinant is associated with the same polypeptide component throughout the purification procedure, indicating that this component is not a result of artifactual aggregation or degradation during isolation. We conclude that the 250,000-dalton polypeptide is part of the saxitoxin binding/sodium channel protein in the native electroplax membrane. PMID- 6280197 TI - Early effects of castration and replacement of androgen on the expression of retrovirus-like particles in rat ventral prostate epithelial cells. PMID- 6280196 TI - Development of chemotaxis and formyl peptide binding in human promyelocytic leukemia cell line (HL60). PMID- 6280199 TI - Collagen synthesis and breakdown in the rat ventral prostate. PMID- 6280198 TI - Increase of plasma neuraminidase activity in experimental peritonitis. PMID- 6280200 TI - Biochemistry of protein kinase reactions in the prostate in relation to androgen action. PMID- 6280201 TI - Photolabeling of benzodiazepine receptors spares [3H]propyl beta-carboline binding. AB - When the benzodiazepine receptor in mouse cerebellar, striatal, and hippocampal membranes was photoaffinity labeled with nonradioactive flunitrazepam, specific [3H]diazepam binding determined with either unlabeled diazepam or ethyl beta carboline-3-carboxylate (betaCCE) as displacer declined greater than 80%. In contrast, specific propyl beta-carboline-3-carboxylate (PrCC) binding in these regions determined with betaCCE as displacer was basically unaltered after photolabeling. Photolabeling lowered specific [3H]PrCC binding with diazepam as displacer to an intermediate extent in the three regions. In cerebellum photolabeling had little effect on either the Kd or Bmax for specific [3H]PrCC binding determined with betaCCE as displacer but significantly lowered the Bmax for specific [3H]PrCC binding determined with diazepam as displacer. These results do not support the idea that [3H]PrCC and [3H]diazepam have a common recognition site on the benzodiazepine receptor. Instead, they suggest that the benzodiazepine receptor is a multicomponent complex. PMID- 6280202 TI - D-Ala2,D-Leu5-enkephalin generalizes to a discriminative stimulus produced by fentanyl but not ethylketocyclazocine. AB - Male Sprague-Dawley rats were trained to discriminate between the effects of saline and either fentanyl (0.04 mg/kg) or ethylketocyclazocine (0.32 mg/kg) by responding on a FR 10 schedule of food reinforcement with a lever on one side of a food cup following a subcutaneous saline injection and responding with a lever on the alternate side following the subcutaneous injection of one of those drugs. The effects of an intracerebroventricular injection of either fentanyl (2.89 14.45 nmol) or D-Ala2,D-Leu5-enkephalin (0.172-1.72 nmol) dose-dependently generalized to the discriminative stimulus produced by fentanyl injected subcutaneously. Intracerebroventricular injection of ethylketocyclazocine (15.7 125.8 nmol) but not D-Ala2,D-Leu5-enkephalin (1.72-13.76 nmol) dose-dependently generalized to the discriminative stimulus produced by ethylketocyclazocine injected subcutaneously. These data demonstrate similarities in the discriminative stimulus properties of proposed micron and delta but not kappa and delta opiate receptor agonists. PMID- 6280203 TI - Effects of naloxone and naltrexone on locomotor activity in C57BL/6 and DBA/2 mice. AB - Administration of naloxone or naltrexone in DBA/2 (DBA) mice was followed by dose related depressant effects. The locomotor activity of the C57BL/6 (C57) mice injected with naltrexone was also depressed. Low doses of naloxone induced a decrease in activity in the C57 strain. This effect gradually disappeared at intermediate doses and recurred again at higher doses. The results are discussed in terms of differences in type number and/or distribution of the receptors influenced by naloxone and naltrexone in the two strains of mice tested. PMID- 6280204 TI - Effects of marihuana cannabinoids on seizure activity in cobalt-epileptic rats. AB - Rats rendered chronically epileptic by bilateral implantation of cobalt into frontal cortices were simultaneously prepared with permanent electrodes for longitudinal recording of the electroencephalogram (EEG) and electromyogram (EMG). Delta-8-tetrahydrocannabinol (delta-8-THC; 10 mg/kg), delta-9 tetrahydrocannabinol (delta-9-THC; 10 mg/kg), cannabidiol (CBD; 60 mg/kg), or polyvinylpyrrolidone (PVP) vehicle (2 ml/kg) was administered IP twice daily from day 7 through 10 after cobalt implantation, at which time generalized seizure activity in non-treated cobalt-epileptic rats was maximal. Relative to PVP treated controls, CBD did not alter the frequency of appearance of seizures during the course of repeated administration. In contrast, both delta-8-THC and delta-9-THC markedly reduced the incidence of seizures on the first and second days of administration. Interictal spiking during this period, on the other hand, was actually enhanced. On the third and fourth days, tolerance to the effect on seizures was evident, with a return of seizure frequency of THC-treated rats to values not significantly different from those of controls. Unlike the effect on seizures, no tolerance developed to the marked suppression of rapid eye movement (REM) sleep induces by delta-8-THC and delta-9-THC. REM sleep remained reduced in the treated animals during the first 2 days after termination of THC administration. In contrast, REM sleep time was unaffected by repeated administration of CBD. These results suggest that delta-8-THC and delta-9-THC exert their initial anticonvulsant effect by limiting the spread of epileptogenic activity originating from the cobalt focus. PMID- 6280205 TI - Antibodies to GM1 ganglioside inhibit morphine analgesia. AB - It has been shown that the injection of antiganglioside serum into the periaqueductal gray matter of rats blocks morphine induced analgesia. This result is due to the action of antibodies to GM1 ganglioside since the specific removal of these antibodies from the antiserum with pure GM1 ganglioside eliminates the blocking activity. Specificity for GM1 ganglioside was further shown by the blocking activity of choleragenoid, which binds specifically to GM1 sites. Antibodies to other brain constituents, namely S-100 protein and myelin, did not block the morphine analgesia. PMID- 6280206 TI - Antagonism of morphine-induced behavioral suppression by opiate receptor alkylators. AB - Experiments were conducted to test the in vivo opiate specificity and long lasting effects of two non-equilibrium opiate antagonists: beta-chlornaltrexamine (beta-CNA) and the beta-fumarate methyl ester derivative of naltrexone (beta FNA). beta-CNA (2.5 or 5.0 micrograms, ICV) partially antagonized suppression of conditioned autoshaped behavior by morphine, when morphine was administered 48-72 hr after beta-CNA. beta-CNA had no effect on amphetamine-induced suppression of autoshaped responding, nor did it antagonize the suppression in rearing activity induced by either morphine or amphetamine. Similarly, beta-FNA (5 mg/kg, IP) antagonized the suppression of conditioned behavior by morphine, for up to 48 hr, while having no effect on amphetamine-induced suppression of autoshaped responding, or on the suppression of rearing activity induced by morphine or amphetamine. Further peripherally administered beta-FNA acts in the brain, since it antagonized analgesia following ICV morphine administration. PMID- 6280207 TI - Effects of chronic phencyclidine on fixed-ratio responding: no relation to neurotransmitter receptor binding in rat cerebral cortex. AB - The effects of chronic phencyclidine (3.2 mg/kg for 25 days) on responding maintained under a fixed-ratio 30 schedule of food presentation were studied in rats. Initially phencyclidine produced large decreases in the overall rate of responding. This decrease was due primarily to long pauses in responding and secondarily to a decrease in local rates of responding. Although tolerance developed to the rate-decreasing effects of phencyclidine in each subject, the extent and pattern of its development differed among the subjects. After the chronic drug regimen, the rats were sacrificed. Ligand binding to muscarinic cholinergic, opiate, adrenergic, and serotonergic receptors in cortex was then compared to that in rats which received saline with operant training, phencyclidine alone, or saline alone. Neither operant behavior alone, phencyclidine alone, nor the interaction of phencyclidine and operant behavior was found to alter binding to these receptors. These results indicate that behavioral tolerance develops to phencyclidine, but it is not accompanied by changes in binding to the receptors studied. PMID- 6280208 TI - Opiate antagonists stimulate affiliative behaviour in monkeys. AB - The effects of treating captive talapoin monkeys acutely (twice daily for 7 days) with naltrexone hydrochloride (0.25 mg 0.5 mg and 1 mg/kg intramuscular injections twice daily), naloxone hydrochloride (0.5 mg/kg IM twice daily) and sulpiride (1.5 mg/kg IM twice daily) was studied in social pairs and singly caged animals. The behaviour of social pairs and endocrine changes in all treated monkeys were monitored before, during and after withdrawal of the course of drug treatment. Naltrexone and naloxone, but not sulpiride, significant increased grooming and grooming invitations while aggressive behaviour, self grooming, scratching and general locomotor activity were unaffected. There was an overall increase in LH, testosterone and cortisol in plasma samples taken 60 mins after opiate receptor blockade. Prolactin was unchanged but increased dramatically in animals treated with sulpiride. No significant endocrine changes were observed to precede the increased grooming behaviour which opiate receptor blockade induced. The behavioural changes reported for this primate support the view that positive affect arising from social bonds may be mediated by cerebral endorphin containing systems. PMID- 6280209 TI - Electrically induced release of (3H)-noradrenaline from rat brain cortex slices: a kinetic analysis of the dependence on extracellular calcium. PMID- 6280210 TI - Pharmacological investigations on electrically-induced changes of cyclic nucleotides in guinea-pig brain slices. PMID- 6280211 TI - Procedures available to examine the immunotoxicity of chemicals and drugs. AB - The discipline of immunology should provide toxicology with sensitive models to assess toxicity as well as provide better safety assessment. More information on correlations between immune function and host resistance changes and a more standardized panel of methods for immunotoxicity screening will add further credence to the already impressive immunotoxicology data base. Research efforts are needed to evaluate newer in vitro methods with microsome activation systems to screen chemicals for immunotoxicity, to evaluate chemicals for induction of hypersensitization or allergy, and to better determine whether there are chemical structure activity relationships for chemically induced immunotoxicity. The studies described here represent preliminary examples of these approaches. PMID- 6280212 TI - Effects of diazoxide and hydrochlorothiazide on water permeability and sodium transport in the frog bladder. AB - The effects of diazoxide and hydrochlorothiazide on vasopressin-induced increments in osmotic water flow and sodium transport across the frog bladder were studied. Diazoxide enhanced the vasopressin-induced osmotic water flow of the bladder, but did not affect the cyclic AMP- or theophylline-induced water flow. Hydrochlorothiazide did not affect the vasopressin-induced water flow. Our results suggest that diazoxide increased the water flow by inhibiting the activity of phosphodiesterase in bladder epithelial cells, whereas hydrochlorothiazide did not. On the other hand, both drugs suppressed the short circuit current of the bladder membrane and inhibited the Na,K-dependent ATPase activity of the kidney cells. These results suggest that both drugs decreased sodium transport in the bladder by inhibiting Na,K-dependent ATPase activity. PMID- 6280213 TI - Proton computed tomography of human specimens. PMID- 6280214 TI - Resolution and contrast recovery at depth in planar nuclear images. AB - Resolution and contrast recovery in planar nuclear images at depth with a high purity germanium (HPGe) camera has been achieved through use of a weighted back projection (WBP) method. The algorithm can be derived from Bayes' theorem using the point spread function of the camera. The variations in the formulation of WBP (one single pass and two iterative procedures) are presented with the characteristics and performance of each method. The evaluation procedure determines the behaviour of signal-to-noise ratio, contrast and texture after application of the algorithm. Both real and simulated cold lesions obtained with the HPGe camera are studied with sizes ranging from 3 mm to 17 mm and background count densities from 100 to 6400 counts cm2. Application of WBP is shown to increase spatial resolution and contrast without a concomitant reduction in signal-to-noise ratio. Images obtained with the HPGe camera and processed with WBP are presented. The algorithm has been applied to the scintillation camera, yielding significant resolution and contrast recovery despite the presence of scatter and textured noise present in the HPGe images. PMID- 6280215 TI - The action spectra for photoreactivation and photorepair in ICR 2A frog cells. PMID- 6280216 TI - Reactivation of UV- and gamma-irradiated herpes virus in UV- and X-irradiated CV 1 cells. PMID- 6280217 TI - Resistance of 3T3 mouse cells to UV light in relation to excision and transfer of dimers to daughter strands. PMID- 6280218 TI - Electron paramagnetic resonance study of storage effects on ceruloplasmin in human serum compared with purified ceruloplasmin in aqueous solution. AB - The EPR signal amplitude of human serum ceruloplasmin shows significant changes as a function of time and temperature during storage. The same behavior occurs with aqueous solutions of purified ceruloplasmin. From the observation that the spectral lines of the EPR signal of ceruloplasmin from unmanipulated serum are identical to those coming from purified ceruloplasmin, we conclude that only type I Cu2+ of ceruloplasmin are involved in the signal changes. A temperature dependent electron shift toward type I Cu2+ paramagnetic centers, occurring via the type II and type III Cu2+ species of the protein, is believed responsible for the process. The possible origin of the reducing electrons is discussed. A procedure to obtain reproducibility of recording of EPR spectra of ceruloplasmin in physiological fluids is proposed. PMID- 6280219 TI - Chemical heterogeneity of structural proteins of cancers with a common low electron spin resonance signal. AB - Electron spin resonance measurements are presented for the separated structural and soluble components of a variety of both normal and cancerous tissues. The signal at g = 2 from the structural part of the normal tissues was at least an order of magnitude stronger than the corresponding signal from the structural part of the cancers. In contrast, no significant difference in radical concentration was found between the soluble fractions of normal and cancerous tissues. Despite their common low signal at g = 2, the structural cancer fractions displayed a remarkable chemical heterogeneity. An important finding was that even cancers of the same histology reacted differently from one another when treated with a variety of chemical probes. PMID- 6280220 TI - Actin polymerization and its regulation by proteins from nonmuscle cells. PMID- 6280221 TI - Organization of visual pathways in normal and visually deprived cats. PMID- 6280222 TI - Responsiveness of the trimmed rat subcutaneous adipose tissue beta-receptors to catecholamines in ontogenesis. AB - In order to estimate the possible changes of the functional state of beta adrenergic receptors during ontogenesis, the lipid mobilizing effects of isoprenaline (ISO), adrenaline (ADR) and noradrenaline (NA) were studied in trimmed subcutaneous adipose tissue of rats at different stages of postnatal development (5, 14, 21 and 35 days after birth. The release of free acids (FFA) into the incubation medium was measured after addition of increasing concentrations of catecholamines and the dose-response curves were evaluated. In all the studied age groups, ISO produced the highest maximum effect and had the greatest potency. The calculated pD2 values indicate the same potency order ISO much greater than NA = ADR in all age groups and they also demonstrate that there exist no appreciable differences in the affinity single catecholamines at different stages of postnatal development. In comparison with all the other groups, the intrinsic activity of the studied adrenomimetics was relatively low in 14-day-old rats. PMID- 6280223 TI - Digitalis-like biological activity and immunoreactivity of synthetic beta 1-24 corticotropin and its natriuretic action. PMID- 6280224 TI - Clinical detection and assessment of drug induced neurotoxicity. PMID- 6280225 TI - Effect of chronic treatment with haloperidol on serum prolactin, striatal opiate receptors and beta-endorphin content in rat brain and pituitary. PMID- 6280226 TI - Anxiety as a state of diminished gabaergic neurotransmission resulting from too frequent recruitment of gabaergic neurons: a neurophysiological model. PMID- 6280227 TI - Regional redistribution of beta-endorphin in the rat brain: the effect of stress. PMID- 6280228 TI - Electroconvulsive therapy update. PMID- 6280229 TI - [Clinical evaluation of the M. v. Ardenne cancer-multistep-therapy concept '74. Part II. Therapy efficiency in inoperable bronchial cancer]. PMID- 6280230 TI - [Intrathecal radiogold application for meningosis "prophylaxis" in children with leukemias and non-Hodgkin lymphomas. II. Results and problems from viewpoint of nuclear medicine]. PMID- 6280231 TI - Changes of acetylcholinesterase activity in different brain areas of the rat following head irradiation. PMID- 6280232 TI - Radioprotective effect of WR-2721 in rabbits: a comparison with cystamine. PMID- 6280233 TI - [Intrathecal radiogold application for meningosis "prophylaxis" in children with leukemias and non-Hodgkin lymphomas. I. Results and problems from the clinical viewpoint]. PMID- 6280234 TI - The value of a portal study in determining the etiology of hepatic masses in the adult. AB - The intrahepatic portal system was studied by superior mesenteric angiography in 80 patients with hepatic masses. The portal study helped to identify the highly significant portal signs of malignancy (amputations, irregular stenosis, portal hypertension), and, among the malignant tumors, the specific signs of hepatoma (portal trunk amputation, ill-defined hepatographic lacunae, association of portal hypertension with proximal portal amputation). It is concluded that the intrahepatic portal study is essential in pretherapeutic staging. It is better than superselective hepatic arteriography in assessing the involvement of one hepatic lobe when a hypervascular tumor involves the other lobe, and it is indispensable if embolization is planned. PMID- 6280235 TI - Redundant lumbar nerve root syndrome: myelographic features. AB - Nine patients with redundant lumbar nerve root syndrome were identified using positive-contrast myelography, seven with metrizamide and two with iophendylate. Serpiginous intradural filling defects occurred in conjunction with a complete or partial extradural block and spinal stenosis. These redundant nerve roots were seen on the cephalic side of the block. Eight of the nine patients had pseudoclaudication. One case was found incidentally during a cervical myelogram. This series is presented to demonstrate the characteristic myelographic pattern that should be recognized and differentiated from tortuous intradural blood vessels. Possible causes are discussed. PMID- 6280236 TI - The Epstein-Barr virus and human lymphoproliferative disorders. PMID- 6280237 TI - [Involvements of cell-to-cell interaction and Na+-Ca2+ exchange system in maintenance of regular beating of cultured heart cells (author's transl)]. PMID- 6280238 TI - [Circadian rhythm in the secretion of corticosteroid and its regulatory mechanisms (authors' transl)]. PMID- 6280239 TI - [The circadian and ultradian related periodicity of pituitary hormone secretion in man and rats (author's transl)]. PMID- 6280240 TI - [Metabolic oscillations and their mechanism; autocatalysis (author's transl)]. PMID- 6280241 TI - A new prostaglandin, C22-PGF4 alpha, synthesized from docosahexaenoic acid (C22:6n3) by trout gill. AB - The concentrations of prostaglandins PGE3 and PGF3 alpha were 214 and 1500 ng/g wet trout gill tissue, respectively. A new prostaglandin, tentatively identified by gas chromatography/mass spectrometry as C22-PGF4 alpha (590 ng/g wet tissue) was discovered. This was synthesized from docosahexaenoic acid. PMID- 6280242 TI - Stimulation of prostaglandin E production by concanavalin A in isolated Graafian follicles. AB - Addition of Concanavalin A (Con A) to isolated rat Graafian follicles induced prostaglandin E (PGE) production after 2 h of incubation. PGE synthesis continued throughout 24 h culture period. Cyclic AMP accumulation was noted after 6 h of incubation with Con A. .Aspirin, indomethacin and flufenamate prevented both the stimulation of PGE production and of cyclic AMP accumulation by Con A; antibodies to PGE prevented the cyclic AMP production. These studies indicate that the interaction of Con A with the follicle results in PGE production. It seems that besides the known pathway for the induction of PGE synthesis in the ovarian follicle, via elevation of intracellular cyclic AMP, an additional pathway, via an external signal which is independent of cyclic AMP exists. PMID- 6280243 TI - Prostaglandin hyperalgesia, IV: a metabolic process. AB - Prostaglandin E2, prostacyclin and Db-cAMP injected into the rat paw induce hyperalgesia. This hyperalgesic effect of the prostaglandins but not of Db-CAMP was blocked by pre-treatment of the animals with cycloheximide. Prostaglandin hyperalgesia thus seems to be dependent on the triggering of some metabolic process which enhances the effects of physical or chemical stimuli. PMID- 6280244 TI - Interactions between parathyroid hormone and prostaglandins on renal cortical cyclic AMP. AB - Effects of parathyroid hormone (PTH) and several prostaglandins (PGs) on cyclic AMP (cAMP) metabolism were studied and compared in isolated renal cortical tubules from male hamsters. Both production and intracellular degradation of cAMP were increased by PTH and each of the PGs tested (PGE2, PGE1, PGI2). Production of cAMP was increased to similar levels by maximal concentrations of PTH and each PG, however, degradation of cAMP was significantly higher in response to PTH than with any of the PGs. This difference in intracellular degradation of cAMP was responsible for the much higher concentrations of cAMP in renal cortical tubules exposed to PGs (PGE1, PGE2, PGI2) than to PTH. Submaximal amounts of each PG produced additive increases in cAMP concentrations in the presence of maximal amounts of PTH. Additivity of the combined responses was lost, however, as the PGs concentrations reached their maxima. The results suggest that renal PGs (PGE2 and PGI2) may modulate the effects of PTH on cAMP concentrations in renal cortical tubules. PMID- 6280246 TI - [Problems of specificity and sensitivity of methods for the rapid detection of antibodies in viral infections]. PMID- 6280247 TI - [Occurrence of enteroviruses in Poland 1973-1979]. PMID- 6280245 TI - Prostaglandins and cancer: a review of tumor initiation through tumor metastasis. PMID- 6280249 TI - Approved HV1 and HV2 systems. PMID- 6280248 TI - Assessments of future environmental trends and problems: applied genetics- agriculture. PMID- 6280250 TI - Certified host vector systems. PMID- 6280251 TI - Case-by-case evaluation of recombinant DNA research involving eukaryotic viruses. PMID- 6280252 TI - Experiments for which containment has been lowered under section III-0-2 of the guidelines. PMID- 6280253 TI - Toxins classified under appendix G of the guidelines. PMID- 6280254 TI - Urinary excretion and circadian rhythm of cyclic-AMP and cyclic-GMP in childhood. AB - Normal urine values of cyclic-AMP and cyclic-GMP were determined in healthy children, using a competitive protein binding and a radioimmunoassay technique respectively. Neither sex nor age affected the cyclic nucleotides excretion. A circadian rhythm in the urinary excretion of cyclic-AMP was observed throughout a 20 h period. The peak excretion occurred at 16.00 h, while minimal excretion was at 24.00 h. The differences observed between the periods of highest and lowest excretion of cyclic-GMP were not statistically significant. PMID- 6280255 TI - The effect of different fiber sources on the neutral steroid excretions of hypercholesterolemic casein fed rabbits. AB - Four groups of 14 New Zealand white rabbits were made hypercholesterolemic by feeding them during four months on a casein semi-synthetic diet containing 21% sawdust. Then 10% sawdust was replaced by cellulose, citrus pectin and wheat bran respectively in three groups, while the fourth group continued on the original sawdust containing diet. A control group consisted of four "chow" fed animals. Excretion of faecal neutral steroids (FNS) was determined periodically. Citrus pectin significantly increased the amounts of FNS excreted after five week. FNS levels were not significantly affected by the other fiber sources, but it tended to be higher on cellulose and wheat bran diets than on the sawdust diet. PMID- 6280256 TI - Cyclic AMP-dependent protein kinase activity and lipolysis in adipose tissue. Effect of fasting, oligomycin and iodoacetamide. AB - The release of glycerol into the medium, the concentration of cAMP, and the cAMP dependent protein-kinase activity were studied in adipocytes and in fat-pads obtained from epididymal adipose tissue of rats under different conditions of feeding. An increase in the tissue concentration of cAMP and in the protein kinase activity was observed in vivo at 48 and 96 h of fasting. A diminished release of glycerol was found in adipocytes from rats fasted for 48 h, in the absence of glucose, and the maximum concentration of cAMP was inferior to that of fed rats. Oligomycin and iodoacetamide, in the presence of epinephrine and glucose, produce a diminution in the values of the parameters studied. No significant differences were observed, however, in the responses of tissue obtained from fed and fasting rats to these compounds. The present results confirm previous observations and show the dependence of the lipolytic process on carbohydrate metabolism. PMID- 6280257 TI - Comparative study of vaccines in preventing respiratory disease in beef calves after weaning. AB - Vaccines against infectious bovine rhinotracheitis (IBR), bovine viral diarrhea (BVD), bovine parainfluenza-3 (PI-3), and bacterin against Pasteurella hemolytica and P. multocida were studied to determine their effectiveness when given 30 days before weaning in preventing respiratory disease in beef calves after weaning. A total of 310 calves, 6 to 8 months old, were divided into 3 groups. Group I consisted of 51 calves vaccinated with a temperature sensitive mutant vaccine intranasally (IBR-PI-3 TSV2). The 56 calves in Group 2 received another intranasal vaccine containing a modified live virus (IBR-PI-3 IP). Calves in both groups received a modified live bovine virus diarrhea (BVD) virus vaccine 30 days before weaning, and a bacteria containing Clostridium chauvei-septicum, Pasteurella hemolytica, and P. multocida. The bacterin was given twice, 2 weeks apart before weaning. The remaining 207 calves were used as unvaccinated controls. All calves were treated at lest once after weaning for clinical respiratory illness. The only virus isolated was PI-3. The serologic response to the viral vaccines was good. There was some doubt as to the effectiveness of the PI-3 component because vaccinated calves were affected by this virus after weaning. Further studies are needed on PI-3 virus vaccine and the most effective vaccination schedule. Vaccination at the time of weaning may have been helpful in the present experiment in preventing clinical disease. PMID- 6280258 TI - Influence of coenzyme A on the in vitro binding of benzo(a)pyrene metabolites to protein. AB - Incubation of 20,000 xg supernatants of liver homogenates from female Wistar rats with benzo(a)pyrene (BP) and other cofactors in the presence of coenzyme A (CoA) produced metabolites that required extensive hydrolysis successively in 2N NaOH, 2N HCl, 2N NaOH and 6N HCl for their release. It is suggested that coenzyme A activated benzpyrenyl-amino acid formed via epoxide(s). The activated amino acid(s) became bound to thiol protein. PMID- 6280259 TI - Intranuclear dense granules in tumour cells of nasopharyngeal angiofibroma. Ultrastructure and submicroscopic cytochemistry. PMID- 6280261 TI - [Diastematomyelia simulating an acrodystrophic neuropathy: diagnosis by computerized axial tomography]. PMID- 6280260 TI - [Embolization in otorhinolaryngology]. PMID- 6280262 TI - Nosocomial infection control: an overview. AB - Nosocomial infections are one of the most important public health problems in the world today. Although a great deal of attention is being directed toward the control and prevention of these infections, old problems continue to occur and new problems are constantly arising. The approximately 5% rate of nosocomial infections among patients admitted to acute-care hospitals has a significant impact on patients morbidity and morality and on the financial concerns of the patient, hospital, and community. An effective, efficient, and sensitive surveillance program developed to meet the needs of the individual hospital is absolutely necessary to obtain data on nosocomial infections from which controls and prevention programs can be developed. At the same time, lessons can be learned from the experience of other hospitals. The nosocomial infection control and prevention program must be under the general direction of a representative, interested, effective infection control committee. One area of importance under the committee's responsibility is that of an appropriate isolation program practiced by all persons within the hospital. PMID- 6280264 TI - [At the fringes of nursing. Heal yourself]. PMID- 6280265 TI - The influence of cyclic 3',5'-adenosine monophosphate on granulation tissue formation. PMID- 6280263 TI - Infectious diseases and immunizations. PMID- 6280266 TI - Responses of plasma adenosine 3',5'-monophosphate, blood glucose and plasma insulin to glucagon in humans. AB - The effect of glucagon on plasma cyclic AMP (cAMP), insulin and blood glucose was examined in normal adult subjects. After an i.v. injection of glucagon there was a rapid, dose-dependent increase of plasma cAMP as well as insulin and blood glucose. Multiple injection of glucagon to the same subject with 60 min intervals gave almost identical responses of plasma cAMP and blood glucose, whereas the insulin response tended to decrease with time. Dose-dependent increases of plasma cAMP, insulin and blood glucose were also seen during a continuous i.v. infusion of glucagon. With the lowest doses of glucagon the blood glucose and plasma insulin concentrations were increased without any change of plasma cAMP. Plasma cAMP, insulin and blood glucose declined prior to the termination of glucagon infusion. During an endogenous hyperglucagonaemia, induced by alanine injection, there was no discernible change of plasma cAMP. We conclude that the early events of glucagon action may be studied in vivo by monitoring plasma cAMP. However, variations of plasma glucagon within the physiological range are not accompanied by measurable changes of cAMP in the peripheral circulation. PMID- 6280267 TI - Recent physiological studies of the alimentary autonomic innervation. AB - The optimum pattern of stimulation of the VIP-ergic fibres in the submaxillary gland of the cat has been investigated by comparing the effects of continuous stimulation at 2 Hz for 10 min with those of stimulation at 20 Hz for 1 sec intervals for the same period. Both the fall in submaxillary vascular resistance (SVR) and release of VIP from the gland are significantly increased when the same total number of impulses is delivered in bursts at the higher frequency. Comparison of submaxillary responses to stimulation in 1 sec bursts, over a wide range of frequencies, has shown that, in atropinised cats, the fall in SVR is linearly related to stimulus frequency over the range 2-60 Hz, and maximal at 80 Hz. In addition, the fall in SVR is linearly related to stimulus frequency over the range 2-60 Hz, and maximal at 80 Hz. In addition, the fall in SVR is linearly related to log VIP output from the gland over the whole of the frequency range 2 160 Hz. Stimulation in 1 sec bursts at 10 sec intervals has also shown that certain non-peptidergic autonomic responses are optimal at much higher stimulus frequencies than has hitherto been supposed on the basis of classical studies employing continuous stimulation. It is concluded that it is no longer justified to assume that autonomic nerve fibres are invariably characterised by low natural discharge frequencies or that they necessarily fire at relatively constant rates. The results obtained using bursts of stimuli also show how differential responses can be obtained in the same tissue simply by varying the stimulus frequency and pattern. PMID- 6280268 TI - Epstein-Barr virus increases the proliferative response and the generation of suppressor and cytotoxic T-cell functions in autologous mixed lymphocyte reaction. AB - Epstein-Barr virus (EBV) infection of stimulator cells significantly increased the proliferative response of T cells in the autologous mixed lymphocyte reaction (AMLR). The addition of a monoclonal anti-HLA-DR antibody to AMLR cultures in which either EBV-infected or non-infected non-T cells were used as stimulator cells strongly inhibited the proliferative response irrespective of the presence of EBV. It is concluded that EBV does not by itself activate the responding cells and that HLA-DR antigens are necessary to trigger T cells. Increased generation of suppressor T cells, determined in both alloantigen-induced DNA synthesis and pokeweed-mitogen-stimulated immunoglobulin production, was found after an EBV infection of stimulator cells. Similarly, EBV-infected non-T cells significantly increased the generation of killer T cells, determined in three different types of target cells: phytohaemagglutinin-stimulated mononuclear cells, EBV transformed cells, and concanavalin-A-activated murine spleen cells. The increased T-cell responses after an EBV infection may reflect the attempts in vivo to control and hold in check the viral infection. PMID- 6280269 TI - Cefotaxime therapy of serious infections with multiresistant gram-negative bacilli. AB - 33 patients with serious gram-negative bacillary infections were treated with cefotaxime. In patients with normal renal function the dose varied between 1.5 to 4 g/day. 17 patients had urinary tract infections, 5 respiratory tract infections, 1 combined urinary tract infection and respiratory tract infection, and 10 miscellaneous infections. 16 patients had septicemia. 25 infections were due to pathogens resistant in vitro to ampicillin, cephalothin, gentamicin and/or tobramycin. 15 infections had failed to respond to ampicillin, cefazolin, gentamicin or tobramycin therapy. 32/33 patients responded favourably to cefotaxime (cure or improvement) but 4 patients developed superinfection with cefotaxime-resistant bacteria. No evidence of nephrotoxicity was observed except for a transient moderate rise in creatinine in one patient. PMID- 6280270 TI - Hemangioma of the lunate (Klippel-Trenaunay syndrome). Case report. AB - An intraosseous hemangioma of the lunate was diagnosed as the cause of wrist complaints. At angiography considerable arteriovenous shunting within the bone was shown. Replacement of the lunate by a SILASTIC implant relieved the pain. The intraosseous hemangioma, however, appeared to be only a part of more wide-spread disease which fulfilled the criteria for a Klippel-Trenaunay syndrome. PMID- 6280271 TI - Granular cell myoblastoma, a rare localization in the trachea. A case report. PMID- 6280272 TI - [The fast and slow channel in myocardium (author's transl)]. PMID- 6280273 TI - [Food fiber and diabetes (author's transl)]. PMID- 6280274 TI - [Contraction-relaxation coupling mechanisms of smooth muscle and calcium regulation (author's transl)]. PMID- 6280275 TI - Solubilized adenosine receptors in the brain: regulation of guanine nucleotides. AB - Adenosine receptors associated with a reduction of adenylate cyclase and labeled by tritium-labeled cyclohexyladenosine can be solubilized from brain membranes with sodium cholate. Regulation of receptor binding by guanine nucleotides is retained in the soluble state. Influences of cations observed in membrane preparations of adenosine receptors are no longer detected with the solubilized receptors. The apparent retention of a complex of receptors and guanosine triphosphate binding but not cation binding protein in the soluble state may permit a molecular analysis of receptor regulation. PMID- 6280276 TI - Spinal sympathetic neurons: possible sites of opiate-withdrawal suppression by clonidine. AB - Morphine, methadone, meperidine, fentanyl, and clonidine rapidly depressed transmission through sympathetic preganglionic neurons in cats with the spinal cord transected. Naloxone promptly antagonized this effect of the opiates but not that of clonidine which was reversed by alpha 2-adrenergic receptor antagonists. The independent depression of preganglionic neurons by clonidine may contribute to the ability of this drug to depress the symptoms of opiate withdrawal that are characterized by sympathetic hyperactivity. PMID- 6280277 TI - Infrared spectrum of the purple membrane: clue to a proton conduction mechanism? AB - The infrared spectrum of the purple membrane of Halobacterium halobium has amide I and amide A frequencies that are anomalously high for standard alpha-helical structures. Normal mode calculations indicate that these and other unusual features of the spectrum can be attributed to alpha 11-helices. Such structures suggest that the helix backbone may provide the framework through which proton transport takes place. PMID- 6280278 TI - Pituitary receptor site blockade by a gonadotropin-releasing hormone antagonist in vivo: mechanism of action. AB - Administration of a potent gonadotropin-releasing hormone (GnRH) antagonist [Nac L-Ala1,pCl-D-Phe2,D-Trp3,6]GnRH as a single subcutaneous injection to castrated adult male rats reduced, by more than 90 percent, both serum luteinizing hormone concentrations and specific pituitary GnRH receptor binding. This effect persisted for 24 hours. The dissociation rate of the antagonist from pituitary membrane homogenates was fourfold slower than the dissociation rate of a potent agonist. The prolonged in vivo inhibition of pituitary GnRH receptor binding and luteinizing hormone secretion by the GnRH antagonist may be mediated by the slower dissociation rate of the antagonist from its specific pituitary membrane receptor site. PMID- 6280279 TI - Experimental hepatic encephalopathy: changes in the binding of gamma-aminobutyric acid. AB - Two populations of receptors for gamma-aminobutyric acid, one with low- and the other with high-affinity characteristics, are detectable in frozen, thawed, Triton-treated synaptic membrane preparations from normal brain. It is now reported that membrane preparations from rats with mild galactosamine-induced hepatic encephalopathy show an increase in the number of low- and high-affinity gamma-aminobutyric acid binding sites, whereas those from rats with severe encephalopathy show only high-affinity binding sites. Thus, hepatic encephalopathy appears to involve partial degeneration of the gamma-aminobutyric acid-containing presynaptic nerve terminals. PMID- 6280280 TI - A new tumor-promoting agent, dihydroteleocidin B, markedly enhances chemically induced malignant cell transformation. AB - Teleocidin, which was isolated from mycelia of Streptomyces, is a potent tumor promoter in mouse skin. The catalytically hydrogenated compound dihydroteleocidin B markedly enhanced malignant cell transformation induced by 3-methylcholanthrene or ultraviolet radiation. Dihydroteleocidin B was at least 100 times more effective in enhancing transformation than 12-O-tetradecanoyl phorbol-13-acetate, the strongest promoter known until now, whereas both promoters showed equal capacities to induce early membrane effects and DNA synthesis. PMID- 6280281 TI - DNA conformation, dynamics, and interactions in solution. AB - The conformation and dynamics of the d(CGCGAATTCGCG) duplex, its analogs containing mismatched base pairs and helix interruptions, and its complexes with actinomycin and Netropsin, bound separately and simultaneously, have been investigated by nuclear magnetic resonance spectroscopy in aqueous solution. Structural information has been deduced from chemical shift and nuclear Overhauser effect parameters, while the kinetics have been probed from line width and saturation recovery experiments on proton and phosphorus markers at the individual base pair level. These studies lead to an improved understanding of the role of nucleic acid sequence on the structure, flexibility, and conformational interconversions in the duplex state. The nuclear magnetic resonance measurements readily identify helix modification and antibiotic binding sites on the nucleic acid and estimate the extent to which the observed conformational and dynamic perturbations are transmitted to adjacent base pair regions. PMID- 6280283 TI - Calmodulin binds to chick lens gap junction protein in a calcium-independent manner. AB - A biochemically active conjugate of calmodulin and tetramethylrhodamine isothiocyanate (CaM-RITC) was synthesized. When incubated with sections of chick lens, this conjugate bound to the surface membranes of lens fiber cells in the presence of absence of calcium. Incubation of lens sections with antibodies to gap junction protein of lens completely blocked the binding of the conjugate to cell membranes, whereas serum from nonimmunized animals or antibodies to others lens proteins reduced the binding only slightly. By means of a gel overlay procedure, 125I-labeled calmodulin was found to bind to the gap junction protein of lens, also in a calcium-independent manner. These results support the concept that calmodulin may interact with and regulate gap junctions in living cells. PMID- 6280284 TI - High angiotensin-converting enzyme activity in the neurohypophysis of Brattleboro rats. AB - The activity of angiotensin-converting enzyme is significantly higher in the intermediate and posterior pituitary lobes of Brattleboro rats than in Long-Evans control rats. The high activity level was reversed by vasopressin treatment. Conversely, angiotensin-converting enzyme activity was significantly lower in the anterior pituitary of Brattleboro rats than in Long-Evans rats, and this activity level was not affected by vasopressin. these findings suggest an inverse relation between vasopressin and angiotensin systems in the posterior and intermediate lobes of the pituitary gland. PMID- 6280285 TI - [Detection and incidence of hepatitis in secondary syphilis. Seventy case-reports (author's transl)]. AB - Biologic hepatitis is common in secondary syphilis. Six cases are reported, in which stereotyped biologic changes were found. These consisted of very high alkaline phosphatase levels and gamma GT levels, with definite inflammation. Cytolysis was moderate or absent. Diagnosis can be established when these biologic anomalies are associated with strongly positive serologic tests for syphilis and rapid recovery under adequate treatment. In these cases, hepatic biopsy is unnecessary. PMID- 6280282 TI - New valiums and anti-valiums on the horizon. PMID- 6280286 TI - [Non-gonadic complications of chemoradiotherapy in Hodgkin's disease. A study of eighty-three patients (author's transl)]. AB - Eighty-three patients with Hodgkin's disease were treated with a combination of chemotherapy and radiotherapy. 43 were included in protocol 1 (from january 1970 to january 1974) and 40 in protocol 2 (february 1974 to december 1977). In protocol 1, staging laparotomy was not systematically performed (20 cases). Treatment consisted of 2 intravenous injections in Vinblastine and total nodal irradiation. In protocol 2, laparotomy was systematic in patients over 50 (35 cases). Patients with stages 1 and II treated as mentioned above. Patients with stage III received two Mopp courses followed by total nodal irradiation. Patients older than 50 with stages I and II and poor prognosis factors received chemotherapy only. Laparotomy was associated with a 0% mortality rate and a 3,6% morbidity rate. No myelitis or pericarditis were observed. Herpes zoster occurred in 24% of the patients, pulmonary apex fibrosis in 6%, hypothyroiditis in 2,4%, and leucopenia in 3,6%. Two late infectious complications were fatal. No solid tumor was apparent. Acute leukemia and non-Hodgkin malignant lymphoma developed in two patients. Good tolerance, shortness of treatment, and remission rate, warrant the pursuit of protocol 2 in which systematic laparotomy for patients under 50 allows total nodal irradiation and therefore reduction of chemotherapy. PMID- 6280287 TI - [Infertility and carbohydrate metabolism. A study of 93 cases (author's transl)]. AB - An oral glucose tolerance test was performed in 93 women with unexplained infertility (sterility or repeated abortions). An abnormal carbohydrate metabolism was found in 1/3 of the cases (32 patients). Advice on diet control, provided to all of these patients, was followed only by 13.9 of these, who conceived less than 6 months after initiation of the low carbohydrate diet, achieved full-term pregnancy. These results suggest a relationship between infertility and glucose intolerance. A glucose tolerance test should be performed in all women presenting unexplained infertility. PMID- 6280288 TI - [Surgical treatment of Prinzmetal variant angina. Coronary artery by pass and plexectomy (author's transl)]. AB - Aorto-coronary by pass, widely accepted in the treatment of patients with coronary artery disease is still controversial in the treatment of Prinzmetal variant angina. Recurrence of attacks, occlusion of grafts and post-operative infarction were frequent by described and seem to be related with the persistence of coronary spasm. Authors propose the association of myocardial revascularization and coronary denervation by resection of periaortic plexi. Results of 50 surgical plexectomies associated with coronary by pass appear far better than by pass alone (mortality 4%, myocardial necrosis 4%, persistence of attacks 4%). PMID- 6280289 TI - [Electromyography in children with polymyositis (author's transl)]. AB - Findings of repeat electromyography (EMG) in 15 children with polymyositis or dermatomyositis are reported. The data classically provided by EMG in these conditions is recalled. Emphasis is put on the contribution of stimulodetection to the diagnosis of primary disease of the muscle. With this procedure, EMG showed myogenic signs in all patients, at the first examination. Electrical anomalies are widespread from the start. A long-term study in 12 patients shows that electrical anomalies remain present long after clinical recovery. EMG disturbances are the last anomalies to disappear. EMG returns to normal when recovery is longstanding. EMG signs reappear if relapse occurs. PMID- 6280290 TI - [Radiological diagrams of the various anatomical forms of mitral incompetency. Report on twenty-nine cases (author's transl)]. AB - The authors compared the radiological findings in 29 cases with the operative or autopsy findings and discuss the criteria at present adopted during standard radiological examination and hemodynamic and angiographic exploration of mitral incompetency to determine the anatomical type. From the radiological findings, one may characterize acute mitral incompetency with a small heart and pulmonary vascular changes from chronic mitral incompetency, the diagnostic criteria of valvular prolapse, ruptures of the chordae tendineae and poor function of the papillary muscles. The authors give a synthesis of their results compared with those in the literature. PMID- 6280291 TI - [Atrial myxoma. Report of eight surgical cases, four with electron microscopy study (author's transl)]. AB - From 1970 to 1979 we studied and operated on 8 patients with atrial myxoma, 7 females and 1 male, 24 to 58 years of age. Symptoms had been present for 4 to 24 months prior to surgery. Five patients were functional class IV (NYHA), 2 class III and one class I. All patients were studied by catheterization and angiography. The diagnosis was suspected from physical findings in 3 patients and was not made before surgery in 2 cases. The tumor was located at the left atrium in 6 patients and the right atrium in 2. Electron microscopy performed in 4 cases revealed the typical findings of myxomatous tissue. Adequate resection under extracorporeal circulation was possible in all subjects. One female who was in severe congestive heart failure died shortly after operation. The survivors did well, remaining functional class I, one to nine years after surgery. PMID- 6280292 TI - [Schmidt's syndrome. Interrelationships between thyroid hormones and cortisol (author's transl)]. AB - An hypothyroidism is discovered. Secondary, a primary adrenal insufficiency is proved. Therefore, we can affirm the Schmidt's syndrome. The anti-adrenal and anti-thyroid antibodies are positive; when the glucocorticoid treatment is begun, the dose of thyroxine treatment must be decreased. That point allows us to discuss the interrelationships between thyroid and adrenals. PMID- 6280293 TI - [Lactic acidosis revealing hepatic carcinoma (author's transl)]. AB - The authors report a case of fatal lactic acidosis which occurred in a 77-year old patient with chronic lymphocytic leukemia. Lactic acidosis revealed a hepatic carcinoma. Pathogenesis and therapy are discussed with reference to previously published data. PMID- 6280294 TI - [Arsenism: a case-report (author's transl)]. AB - Among the multiple clinical features encountered in arsenism the most striking are hematological (leucopenia, macrocytic anemia), neurological, digestive, and cardiovascular. Diagnostic rests on the assay of the arsenic content of hair and urine. D-penicillamine appears to be the best therapy. However, late development of cancer of the skin or of internal organs remains possible. The authors report a case of arsenism and review the aspects of this rare disease. PMID- 6280295 TI - [Pseudotuberculous hydatidosis of the peritoneum. Report of an autochthonous case (author's transl)]. PMID- 6280296 TI - [Is there any justification to the scintigraphy and echotomography B association for the tracking down of hepatic metastases? (author's transl)]. AB - Regarding the tracking down of hepatic metastases, a study was made on 58 patients presenting either digestive or genital adenocarcinoma. The determining criterion of selection was the carrying out of the three following tests within the same month (Technetium 99 Scintigraphy, Echotomography B and surgery). The global breakdown of exact answers is in favour of echography (72% against 53%), echography being more reliable than scintigraphy. Therefore echotomography B seems to be the first morphological method that should be proposed; and for improving the tracking down, scintigraphy makes the interpretation of the results more difficult, and reduces the number of correct answers. PMID- 6280297 TI - [Pharmacodynamics of alizapride (author's transl)]. AB - Studies of alizapride (N[(allyl-1 pyrrolidinyl-2) methyl] methoxy-2 azimido-4,5 benzamide hydrochlorate) in mice and rats demonstrated little toxicity, particularly after parenteral administration. Alizapride's main pharmacodynamic effects are on the central nervous system. It is very effective against emesis induced by apomorphine and dihydrogenated ergot alkaloids in dogs. In this respect it is three times more effective than metoclopramide. In contrast to neuroleptics, alizapride does not modify equilibrium reflexes in mice, nor does it reinforce hypnosis induced by barbiturates. Only minor central antidopaminergic effects were recorded, less marked than those seen with metoclopramide. In mice, alizapride has no anticonvulsant or analgesic effects. It has little action on the autonomic nervous system or on the cardiovascular system. Alizapride has no antihistaminic or parasympatholytic effect. In dogs, sympatholytic effects and hypotension are seen only after giving a much higher dose than that which is effective against apomorphine and dihydrogenated ergot alkaloids. PMID- 6280298 TI - [A study of cardiovascular and antiarrhythmic effects of alizapride (author's transl)]. PMID- 6280299 TI - [Pharmacokinetics and bioavailability of alizapride (author's transl)]. PMID- 6280300 TI - [A comparative study of alizapride and metopimazine (author's transl)]. PMID- 6280301 TI - [A study of the effectiveness of alizapride against emesis in children (author's transl)]. PMID- 6280302 TI - [A study of alizapride, a new benzamide, in nausea and emesis in internal medicine (author's transl)]. PMID- 6280303 TI - [Effectiveness of alizapride against emesis in infants and children (author's transl)]. AB - The effectiveness of alizapride against vomiting was studied in a group of 40 patients including neonates, infants and children. Patients were hospitalized in a surgical ward. 2 to 8 mg/kg/24 h were given orally. Vomiting ceased in 75% of the cases. Optimal results were obtained with 4 to 5.6 mg/kg/24 h. With alizapride symptoms abated in all patients. Operative conditions were improved for those who needed surgery. In addition, nausea was relieved in 100% of cases and regurgitations in 67.7%. Our study shows that alizapride is a reliable drug with a short time lag and no side-effects. We conclude that it can be used against vomiting in pediatric patients. PMID- 6280304 TI - [A double-blind crossover trial of alizapride versus placebo regarding effectiveness against nausea and emesis induced by cancer chemotherapeutic agents (author's transl)]. PMID- 6280305 TI - [A study of the antiemetic effect of alizapride (author's transl)]. PMID- 6280306 TI - [A double-blind study of alizapride in nausea and emesis induced by cancer chemotherapeutic agents (author's transl)]. PMID- 6280307 TI - [Granular cell myoblastoma (Abrikosov tumor) of the esophagus (author's transl)]. AB - Granular cell myoblastoma (Abrikossoff tumor) of the esophagus is exceptional: seventeen cases have been described. We report two new observations. They were both unexpectedly discovered during fiberoptic endoscopy performed for gastric symptoms. Diagnosis was established only by histological examination. PMID- 6280308 TI - [Comparison of various methods of diagnosis of the mitral valve (author's transl)]. AB - This study compares the information supplied by auscultation, echocardiography and left ventriculography in the diagnosis of mitral prolapse from a study of the world literature and a personal study. A click on auscultation or a telesystolic click murmur is noted in almost 74 p. cent of cases. Echocardiography may lead to false negatives in partial prolapse of the posterior valve and in ventriculography, even judged according to strict and restrictive criteria they may make errors by excess. Telemetric echocardiography may assess in an overall way the number or prolapsed valvular structures and the degree of mitral incompetence. The respective indications of the 3 methods are given in detail. PMID- 6280310 TI - [Mac Leod syndrome]. AB - The Mac Leod syndrome is a unilateral pulmonary hypoplasia with emphysema. Onset usually occurs in childhood. The authors delineate this well-defined nosologic entity which is one of the many etiologies of the unilateral hyperlucent lung. PMID- 6280309 TI - [Primitive ano-rectal neuralgia. Atypical cases (author's transl)]. AB - In a new statistic concerning 245 cases of primary ano-rectal neuralgia, it can be seen that the schematic classical classification derived from Thaysen, Theile, and Bensaude must be maintained (proctalgia fugax: 45 cases; coccygodynia: 44 cases; ano-rectal neuralgia: 95 cases), partially integrating unusual cases of pruritus ani. Among the 4 principle etiologies, while not underestimating the neuropsychical and intestinal factors (constipation, laxatives), the importance of two other factors must be underlined (the urogenital factor, and particularly, the role of menopause, and important pelvic operations (33 cases) often overestimated; rachidian factors: tendomyositis (Garrigues), pseudoradicular factor. The interest of this study is to show that besides these typical cases (81,7%), a number of atypical cases exist, which have often been under-estimated. These cases can be classified in intermediary (4%), associated (10%), alternating (3,3%) cases, in the course of which the different syndromes replace each other or seem superposed. It must be underlined that the notion of these primitive ano rectal neuralgias must be inserted in the much larger class of perineal urinary, gynecological or bone and ligament neuralgias. The classification remains opened. An etiopathogenical treatment must be installed, that rejects all regional or surgical aggressive acts when not absolutely necessary, and underlines the importance of massage, internal (levator ani), or external (Maigne's technique, attentive and repeated sessions of rachidian massage).U PMID- 6280311 TI - [Cerebral ischemia after ligature of both carotid arteries in rats: effect of ginkgo biloba extracts (author's transl)]. AB - Simultaneous ligature of both carotid arteries in Long Evans rats results in death for all animals. 50% of the animals survive if the ligatures are separated by a four-day interval. Studies of variations in cerebral blood flow according to the time interval between the two ligatures show that alternative vascularization develops and that the balance between both hemibrains is restored. After giving L dopa3H to animals with ligature of both carotids, increased synthesis of cerebral dopamine 3H and norepinephrine 3H can be demonstrated. When animals are given extracts of Ginkgo biloba leaves prior to carotid ligatures, survival rate is improved and dopamine synthesis increased, without marked changes in cerebral blood flow. This is related to increased brain cell activity in spite of hypoxia due to carotid ligature. PMID- 6280312 TI - [Supra-selective vagotomy in the treatment of perforated duodenal ulcers. Thirty cases report (author's transl)]. AB - Thirty patients with acute perforated duodenal ulcer were managed by supra selective vagotomy and simple closure of the perforation. There was no operative mortality and a low morbidity. Fourteen patients have had gastric studies between four and thirty-two months after operation. Three patients had transitory epigastric fullness (21%); there was no diarrhea or dumping, but two recurrences (14%). The risk of ulcer recurrence after simple closure is high; supra-selective vagotomy as a definitive ulcer therapy, does not increase operative mortality, and has mild digestive sequelaes, for an acceptable rate of recurrence. PMID- 6280313 TI - [A clinical trial of veralipride in menopausal complaints (author's transl)]. AB - Menopausal complaints, especially hot flushes, are common occurrences. Until now, no effective treatment was available for patients in whom estrogens are contraindicated. A trial of veralipride was carried out in ten symptomatic menopausal women (7 physiologic menopauses, 3 surgical menopauses). Symptoms included severe hot flushes, anxiety (N = 6), depression (N = 4), irritability (N = 3), and cephalalgia (N = 2). Patients were given one tablet of veralipride each day for twenty day periods. Effectiveness on hot flushes was excellent in six patients, satisfactory in two, mediocre in one, and null in one. Depression, anxiety, and, to a lesser extent, cephalalgia and irritability, were alleviated. No clinical or biological adverse side-effects were recorded. PMID- 6280314 TI - [Avascular vertebral necrosis in adults (author's transl)]. AB - A new case of avascular vertebral necrosis in a sixty-five-year-old woman is reported. Roentgenograms showed an intervertebral vacuum cleft leading to unequivocal diagnosis. The main features of vertebral necrosis are recalled. PMID- 6280315 TI - [A new case of carcinoma arising in Caroli's disease (author's transl)]. AB - A new case of carcinoma involving Caroli's disease in a 51 years old woman is reported. This papillary carcinoma is developed in left hepatic duct. The type is quite different from the previous cases already described. It is not a cholangiocarcinoma but a papillary mucoid carcinoma. PMID- 6280316 TI - [Factor X deficiency in amyloidosis (author's transl)]. AB - Constitutional aplasia of the bone marrow is a general term which designates familial aplastic pancytopenias with recessive autosomal transmission. They are rare and often associated with malformations which are of great diagnostic value. The Fanconi syndrome is a well individualized form in which aplasia of the bone marrow is combined with various malformations which are of great diagnostic value. The Fanconi syndrome is a well individualized form in which aplasia of the bone marrow is combined with various malformations, particularly skeletal, cutaneous and renal. Cytogenic lymphocyte analysis shows an abnormal tendency to chromosomal breakage which is increased after incubation with alkylating agents. Bone marrow transplantation is the only available therapy for these constitutional aplasias. PMID- 6280317 TI - [Constitutional aplastic pancytopenia in children (author's transl)]. AB - Constitutional aplasia of the bone marrow is a general term which designates familial aplastic pancytopenias with recessive autosomal transmission. They are rare and often associated with malformations which are of great diagnostic value. The Fanconi syndrome is a well individualized form in which aplasia of the bone marrow is combined with various malformations which are of great diagnostic value. The Fanconi syndrome is a well individualized form in which aplasia of the bone marrow is combined with various malformations, particularly skeletal, cutaneous and renal. Cytogenic lymphocyte analysis shows an abnormal tendency to chromosomal breakage which is increased after incubation with alkylating agents. Bone marrow transplantation is the only available therapy for these constitutional aplasias. PMID- 6280318 TI - [Diagnosis of fever in a patient returning from black Africa (author's transl)]. AB - The diagnostic problems which arise when fever occurs in a patient returning from black Africa are more and more frequently encountered because of the multiplication of rapid connections with this continent. Analysis of the main etiologies leads the author to review most of the specifically tropical diseases. However, cosmopolitan diseases should not be underrated. The author emphasizes the high incidence of pernicious malaria, liver amebiasis, and typho-paratyphoid fevers, along with the necessity of keeping in mind the new viral diseases (Lassa, Marburg, Ebola). PMID- 6280319 TI - [Acute paracetamol poisoning. A retrospective study of data from the Paris Antipoison Center. 1974-1981 (author's transl)]. AB - Seventy-five cases of acute poisoning with effervescent tablets of paracetamol have been recorded. No patients developed hepatitis. Ingested doses were low, and resulted only in minor manifestations. As each package contains only 6.6 g of paracetamol and effervescent tablets cannot be ingested in large quantities, the risk of serious overdosage is low. PMID- 6280320 TI - Neurotoxicity of antineoplastic drugs. PMID- 6280321 TI - The newborn with poor muscle tone. PMID- 6280323 TI - Some miscellaneous joint diseases. PMID- 6280322 TI - Calcium deposition diseases. PMID- 6280324 TI - Multiple granular cell myoblastomas of the bronchial tree. PMID- 6280325 TI - Plasma cell myeloma simulating Krukenberg's tumor. PMID- 6280326 TI - Otogenic pasteurella multocida brain abscess and glomus jugulare tumour. AB - We report the occurrence of a Pasteurella multocida temporal lobe abscess in an elderly woman who had a history of neglected chronic purulent otitis and in whom an extensive ipsilateral glomus tumour invading the petrous bone was found. We believe this is the first report in the literature of an otogenic cerebral abscess associated with a glomus jugulare tumour and the fifth report of a Pasteurella multocida brain abscess. The synergistic pathogenesis of the otitis and the glomus tumour in the evolution of the abscess is hypothesized. PMID- 6280327 TI - [Neurogenic tumors of the nasal cavity and paranasal sinuses]. PMID- 6280328 TI - [Rotavirus gastroenteritis (clinical course and diagnosis)]. PMID- 6280329 TI - [Prazosin--pharmacology and mechanism of action]. PMID- 6280330 TI - Gallium-67 in the evaluation of sarcoidosis: correlations with serum angiotensin converting enzyme and bronchoalveolar lavage. AB - Gallium-67 (67Ga) scanning was assessed for its usefulness in the evaluation and follow-up of 54 patients with sarcoidosis, both treated and untreated. Scans were repeated in 23 subjects. Serum levels of angiotensin-converting enzyme (ACE) were determined concurrently in all 54 patients and bronchoalveolar lavage was performed in 29 patients. Gallium-67 scan was effective in the detection and assessment of lesions not revealed by traditional methods of investigation, particularly those affecting the mediastinum, spleen, and salivary glands. The scan also enabled fibrotic lesions, which do not show uptake, to be distinguished from granulomatous lesions, which do--an advantage of prognostic interest particularly in patients with pulmonary lesions. Another merit of 67Ga scanning was that it offered a means of following disease progression in each site. In patients showing spontaneous clearing of disease or receiving treatment the scintigraphic method was more sensitive than serum ACE determination. Scan findings showed a rough correlation with serum ACE but not with bronchoalveolar lavage findings. This suggests that the three markers probably reflect different stages of the granulomatous process. On the strength of this study the indications for gallium scanning in sarcoid patients can be defined more clearly than has previously been possible. PMID- 6280331 TI - Lung inflammation in sarcoidosis: comparison of serum angiotensin-converting enzyme levels with bronchoalveolar lavage and gallium-67 scanning assessment of the T lymphocyte alveolitis. AB - Serum angiotensin-converting enzyme (ACE) is elevated in many patients with pulmonary sarcoidosis and has been proposed as a measure of disease activity. The present study was designed to evaluate the possible relationship between serum ACE and direct measures of the intensity of the alveolitis of pulmonary sarcoidosis as measured by bronchoalveolar lavage and gallium-67 (67Ga) scans. To accomplish this, 64 measurements of serum ACE, lavage T lymphocytes, and lung uptake of 67Ga were performed in 41 patients with biopsy-proven sarcoidosis. Elevations of serum ACE were found on at least one occasion in 17 patients (41%). However, serum ACE was found to be a poor predictor of the intensity of alveolitis in sarcoidosis as assessed by the quantitation of bronchoalveolar lavage cells that were T lymphocytes and by 67Ga scanning. Elevated serum ACE did not predict which patients would have elevated proportions of lavage T lymphocytes, which patients would demonstrate increased pulmonary uptake of 67Ga, or which patients would have high-intensity alveolitis as defined by a combination of these criteria. These observations suggest that while serum ACE may be useful in diagnosing sarcoidosis, it does not reflect accurately the intensity of the alveolitis of the pulmonary component of this disease. PMID- 6280333 TI - The effect of tetracosactrin on platelet aggregation in patients with primary adrenocortical failure. PMID- 6280334 TI - [Clinical evaluation of intravenous mecillinam in patients with chronic urinary infection and reduced renal function]. PMID- 6280332 TI - Clinical and biological activity of the antiplatelet agent suloctidil in treatment of idiopathic recurrent vein thrombosis (I.R.V.T.). AB - A double blind cross over study with suloctidil (Sulocton, Continental Pharma) and placebo was carried out for 6 months in 31 patients with idiopathic recurrent vein thrombosis. They were previously unsuccessfully treated with vitamin K antagonists (VKA) (18 patients) or acetylsalicylic acid (ASA) (13 patients) combined or not the a fibrinolysis activator (theophylline nicotinate). Clinical features, ultrasonic venous flow and biological parameters were controlled monthly during the 6 month treatment. Relevant improvement of clinical, ultrasonic and biological parameters was only observed under suloctidil therapy: during placebo administration 12 patients developed new thrombotic events complicated in 2 by pulmonary embolism while none occurred under suloctidil therapy. PMID- 6280336 TI - [Diagnosis of insulinomas. Determination of insulin, C-peptide and proinsulin in blood to differentiate between endogenous and exogenous hyperinsulinism]. PMID- 6280335 TI - [Benzodiazepine receptors]. PMID- 6280337 TI - HLA antigens in patients with recrudescent herpes simplex infections. AB - We have assessed the humoral and cellular immune responses to HSV-1 antigen in normal controls, and in 47 subjects with recurrent cold sores. THe antigens at the HLA-ABC and DR loci were examined in the first group of 21 susceptible subjects. An apparent increase in prevalence of A9 seen in this group was confirmed in a prospective study of a further 26 subjects (P less than 0.0005). In contrast to previous studies, no relationship was noted between the immune responses and HLA antigens. PMID- 6280338 TI - Nature of serum ascorbate radical and its quantitative estimation. AB - A doublet signal was observed in human serum by the ESR technique at room temperature. This radical had a g value of 2.0054 and a hyperfine splitting constant of 1.84 gauss and was assigned to ascorbate radical. The ascorbate radical in serum was very stable. The intensity of ESR signal showed no differences between serum and plasma of the same individual. Photosensitivity of the ascorbate radical in serum and sodium ascorbate solution was examined and enhancement of ESR signal by irradiation was observed, although the responses in serum and in ascorbate solution were considerably different. The intensity of ESR signal was proportional to the concentration of ascorbate solution. The ESR intensity of ascorbate radical can be used as a reliable method for quantitative estimation of ascorbate radical. PMID- 6280339 TI - Chemiluminescence of UV-irradiated linolenic acid and squalene. AB - An emission spectral analysis was carried out on ultraweak chemiluminescence emitted from UVB-irradiated linolenic acid and squalene. The main emission species produced by the transition of (1 delta g) (1 delta g) dimer and an additional weak band near 477.5 nm (0, 0) by the transition of (1 delta g (1 epsilon g+) to (3 epsilon g-) (3 epsilon g-) were found by spectroscopic analysis of chemiluminescence in both cases of irradiated linolenic acid and of squalene. A distinct peak around 410-420 nm was observed in irradiated squalene and the emitter seems to be due to the excited carbonyl compound. PMID- 6280340 TI - Acute pathologic effects of 3,3',4,4',5,5'-hexabromobiphenyl in rats: comparison of its effects with Firemaster BP-6 and 2,2',4,4',5,5'-hexabromobiphenyl. PMID- 6280341 TI - D-Penicillamine toxicity in mice. II. Concentrations of Cu, Zn, and Fe related to development of toxicity. PMID- 6280342 TI - Effects of polybrominated biphenyls on metabolism of testosterone by rat hepatic microsomes. PMID- 6280343 TI - Inhibition of cholinesterase activity by isocyanates. PMID- 6280344 TI - A simple method for the isolation of vitamin D metabolites from plasma extracts. AB - A simple method has been developed using 'SEP-PAK' disposable silica cartridges to separate the major endogenous vitamin D metabolites, namely vitamin D3, 25 hydroxy vitamin D3 (25OHD3), 1,25 dihydroxy vitamin D3 (1.25 (OH)2D3) and 24,25 dihydroxyvitamin D3 (24,25 (OH) 2D3). After extraction of plasma in isopropanol toluene (25:75) the dried extract is reconstituted in hexane; this is applied to a SEP-PAK column, and stepwise elution carried out under gravity with 0.1 divided by isopropanol in hexane (neutral lipids), 1% isopropanol in hexane (D3), 3 divided by isopropanol in hexane (25OHD3), 3.125 divided by ethanol in dichloromethane (24,25 (OH) 2D3) and 50 divided ethanol in toluene (1, 25(OH) 2D3). Complete separation of these D3 metabolites is achieved by this process and up to 40 samples can be handled at one time. If combined with a suitable ligand binding assay, the system appears to be suitable for preparation of samples prior to the routine assay of vitamin D metabolites. PMID- 6280345 TI - Dopaminergic control of aldosterone: modulation of the response of rat adrenal zona glomerulosa cells to alpha-Msh by pretreatment with bromocriptine or metoclopramide. AB - Bromocriptine treatment in rats (3 mg/kg per day, 7 days) significantly reduced alpha-msh and aldosterone plasma levels 2 hrs after the final treatment in animals on low, normal and high sodium diets. Alpha-MSH dose response curves for corticosterone and 18-hydroxydeoxycorticosterone (18-OH-DOC) in subsequently incubated glomerulosa cells gave stimulation at lower concentrations of alpha-MSH (10(-10) moles per litre) than in cells from untreated animals (10(-9) moles per 1). Curves for aldosterone (ald) and 18-hydroxycorticosterone (18-OH-B) were also affected in cells from animals on a low sodium diet. Fasciculata-reticularis cell responses to ACTH were unaffected. Metoclopramide (4 mg/kg per day, 7 days) elevated plasma alpha-MSH, although ald was unaffected, but inhibited the glomerulosa cell response to alpha-MSH in vitro. Acute dopaminergic responses in plasma ald may be mediated through alpha-MSH in rats, but chronically alpha-MSH may down- regulate glomerulosa cell alpha-MSH receptors. It is unlikely that alpha-MSH mediates the adrenocortical response to sodium depletion. PMID- 6280347 TI - Effects of long-term intake of DDT on rats. AB - DDT is a pesticide used in malaria-control programmes throughout the world. Its potential carcinogenicity was studied in MRC Porton rats (Wistar-derived) which received dietary concentrations of 0, 125, 250 and 500 parts per million DDT (technical-grade) for life. The treatment had no adverse effects on body growth or survival rate. Various types of tumours were observed in animals in all groups: exposure to DDT resulted in statistically significant increased incidence of liver-cell tumours only in female treated rats; one such tumour was observed in control rats. No metastases of these tumours were found. PMID- 6280346 TI - [Effect of thallium on enzyme activity and unidirectional sodium transport in frog skin]. AB - The cytophotometric analysis of enzyme activity in the frog skin epithelium has shown that Tl+ accumulated by cells at millimolar concentrations causes a 70-80% inhibition of both succinate and alpha-ketoglutarate dehydrogenases, while the activity of Na, K-ATPase decreased only slightly. The opposite situation was true for the ouabain treatment. The accumulation of Tl+ by frog skin caused a substantial swelling of mitochondria. It is suggested that the earlier observed inhibition of the unidirectional Na+ transport by Tl+ might be resulted from a blocking of oxidative metabolism. The same cells poisoned by Tl+ were able to maintain their ion composition presumably at the expense of glycolysis. PMID- 6280348 TI - [Association of laparoscopy and bone marrow biopsy in the staging of small-cell undifferentiated carcinoma of the lung. Evaluation of 116 consecutive cases]. AB - 116 consecutive patients with small cell anaplastic carcinoma of the lung had laparoscopy with multiple liver biopsies and bone marrow biopsy, performed as a staging procedure. Bone marrow biopsy was positive in 13 patients (11.2%); no significant rise in percent was seen when bilateral specimens were obtained. Laparoscopy was positive in 17 patients (14.6%); one patient had extrahepatic involvement only. Liver scan did not show high correlation with laparoscopy: 6 patients out of 11 had negative liver scan with metastases demonstrated at laparoscopy. Only 6 patients had laparoscopy positive along with bone marrow biopsy (5.1%). At determination of the stage, 12 patients out of 78 (15.3%), with apparently localized disease had diffuse disease after the two procedures. Laparoscopy and bone marrow biopsy were positive in 12 patients out of 38 (31.5%), when the disease was diffuse at diagnosis. 7 patients had liver metastases at diagnosis, which were excluded after laparoscopy. Laparoscopy and bone marrow remain valuable procedures in assessing stage in small cell anaplastic carcinoma of the lung, because of their high rate of stage modifications due to their combined employment at diagnosis. PMID- 6280349 TI - Electron microscopic observations on cultured fibroblasts from Fabry heterozygotes and hemizygotes. AB - The ultrastructure of cultured fibroblasts from 2 hemizygotes with Fabry's disease ad 8 heterozygotes was compared with that of normal cells. Lysosomal inclusions consisting of tightly packed, concentric lamellae were seen in all fibroblast lines carrying the Fabry gene, whether hemizygous or heterozygous, but not in normal fibroblasts. Most heterozygous, but not in normal fibroblasts. Most heterozygous fibroblasts have fewer lysosomal inclusions than the hemizygous cells. The morphology of the inclusions is similar to that seen in biopsy material. Among the heterozygotes no correlation was found between enzyme levels and fibroblast inclusions. PMID- 6280350 TI - [Alloplastic grafting of ligaments with carbon-fibres (author's transl)]. AB - During several years ligaments consisting of carbon-fibres have been tested in animal experiments. Recently this material was used in humans. Until today approximately 150 patients have been operated. The late results in 65 patients with instability in 66 joints are reported. Carbon-fibre-ligaments yielded good results, provided an expert surgical technic was applied and the follow-up care was optimal. Good results were achieved on the shoulder, the knee and the lateral ankle-joint. Initially there arose some problems due to technical difficulties. By using a modified surgical technic and improvements in the follow-up care still better results can be achieved. The present study is retrospective; a prospective study has been started. PMID- 6280351 TI - [Surgical fracture fixation, development, efficacy and results compared with conservative methods--a report]. PMID- 6280352 TI - Fast hardening aminoacid-solution as potential carrier of chemotherapeutic agents in the treatment of chronic osteomyelitis. AB - Report about an aminoacid, which hardens quickly in liquid environment. It is highly suitable as carrier of chemotherapeutic agents. In-vitro tests show an antibiotic inhibitory activity of different compounds for at least 41 days. According to in-vivo experiments in 17 rats the new substance is replaced in the bone and in soft tissues by fibrous tissue; the antibiotic activity is released slowly. This offers a new carrier which is suitable for occlusion of cavities. The instillation of the new amino-acid (Ethibloc) combined with antibiotics could be a new treatment for chronic osteomyelitis. PMID- 6280353 TI - Antegrade catheter technique to dissolve uric acid ureteral calculi. PMID- 6280354 TI - Case profile: double-contrast urethrography for visualizing small lesions in distal urethra. PMID- 6280355 TI - [Concentration of blood adrenocorticotropic hormone and cortisol in patients with acute pancreatitis]. PMID- 6280356 TI - [Fibroadenoma phyllodes of the breast]. PMID- 6280357 TI - [The effect of feeding sawdust and fodder cut into various lengths on rumen fermentation]. AB - Wethers were subjected to experiments aimed at studying the possibility of replacing part of bulk fodder by treated beech sawdust and its effect on ruminal fermentation in the case of administration of chopped and unchopped hay. For a period of 24 weeks the animals consumed 1000 g of diet dry matter daily; the diet contained 41.81% of chopped meadow hay, 25.28% of barley, 15.37% of chopped straw, 14.98% of molasses, 1.32% of urea and 1.24% of mineral supplement. In the test group the 15.37% proportion of straw was replaced by sawdust. In the subsequent ten-week period the animals were given the same diets, but the hay was not chopped. On the whole the experiment was performed with two groups with straw and two with sawdust. In the groups with chopped hay the sawdust in the diet increased the molar percentage of acetic acid, reduced the molar percentage of propionic acid (which resulted in an increased acetate: propionate ratio), and reduced the pH value and energy efficiency of the produced volatile fatty acids (VFA). The levels of total VFA were not affected. In the groups fed unchopped (long) hay the diet with 15.37% of sawdust showed a significant increase in total VFA and in the molar percentage of butyric acid as well as in the energy efficiency of VFA in rumen. The acetate: propionate ratio was not changed, probably owing to sufficient production of acetic acid. Although some variability was recorded in the results, it can be stated that treated beech sawdust had no adverse effect on rumen fermentation and that the replacement of part of bulk forage by this sawdust was successful. A relationship was also found between sawdust and the physical form of the bulk fodder contained in the ration. PMID- 6280358 TI - The effect of lithium chloride on renal structure and sodium-potassium-adenosine triphosphatase activity in dogs. AB - Lithium chloride was given intraperitoneally to dogs at a dosage of 125 mg/kg body weight for three days. Kidneys were removed for morphologic examination and quantitation of sodium-potassium-adenosine triphosphatase (Na-K-ATPase) activities in cortical and medullary tissue. Light microscopy showed no changes in the kidneys, but cytoplasmic vacuolation and dilatation of the cisternae of the endoplasmic reticulum were seen ultrastructurally in the epithelial cells of the distal tubule and cortical and medullary collecting ducts. Mean cortical Na-K ATPase activity was 1.49 +/- 0.25 and 1.70 +/- 0.31 mumoles inorganic phosphate/mg protein/hour in control and experimental groups respectively. Mean medullary Na-K-ATPase activity was 4.71 +/- 0.41 and 5.01 +/- 0.47 mumoles inorganic phosphate/mg protein/hour in control and experimental groups respectively. It was concluded that lithium produced morphologic changes in the distal nephron, but had no effect on renal Na-K-ATPase activity. PMID- 6280359 TI - Pathology of experimental CV777 coronavirus enteritis in piglets. I. Histological and histochemical study. AB - Sixteen cesarean-derived colostrum-deprived piglets were infected oronasally with CV777 coronavirus on the second or third day of life. Two uninfected piglets were controls. They were killed at 96 and 120 hours after birth. After an incubation period of 22 to 36 hours, all principals showed severe diarrhea. The principals were killed between 12 and 120 hours after infection. Exfoliation of enterocytes were seen first in the piglet killed 24 hours after infection (two hours after the diarrhea began). From that time on, shortening and fusion of villi was present in all small intestinal parts. Affected cells showed vacuolation. The histochemical study showed that infected piglets had decreased activity of all four enzymes studied. The light microscope showed no lesions in the absorptive colonic epithelium. The significance of the lesions in relation to intestinal dysfunction is discussed, and lesions are compared to those of transmissible gastroenteritis and porcine rotavirus infection. PMID- 6280360 TI - Pathology of experimental CV777 coronavirus enteritis in piglets. II. Electron microscopic study. AB - Sixteen cesarean-derived, colostrum-deprived piglets were infected oronasally with CV777 coronavirus on the second or third day of life. Two uninfected piglets were controls. After an incubation period of 22 hours to 36 hours, all principals showed severe diarrhea. The piglets were killed at different time intervals. Viral particles were found in the jejunal villous epithelial cells from 18 hours after infection until four days after the beginning of diarrhea. In the colonic epithelial cells, viral particles and degenerative lesions were found only in the piglet killed 36 hours after onset of diarrhea. Degenerative lesions in the enterocytes began at 18 hours after infection and were most pronounced in the jejunum at the onset of clinical signs. From 24 hours on after the onset of clinical signs, three cell types were found: degenerated virus-containing enterocytes; cuboidal cells; and columnar, highly vacuolated cells containing lipid droplets. PMID- 6280361 TI - Acute encephalitis and hydrocephalus in dogs caused by canine parainfluenza virus. AB - Gnotobiotic puppies were inoculated intracerebrally with a strain of canine parainfluenza virus (CPI-78-238). Four of eight dogs developed histological evidence of acute encephalitis. Clinical signs of encephalitis were seen in two of these four dogs; one had signs and lesions of interstitial pneumonia. Of six inoculated dogs observed for six months after infection, five developed internal hydrocephalus. Virus was reisolated from two dogs with acute encephalitis but not from dogs with hydrocephalus. Hemagglutination-inhibition antibodies persisted throughout the observation period of six months at high levels in the serum and cerebrospinal fluid of hydrocephalic dogs. PMID- 6280362 TI - Multicentric bronchiolo-alveolar neoplasm in a steer. PMID- 6280363 TI - Sequential pathological study of experimental porcine adenovirus enteritis. AB - The sequential enteric lesions of experimental porcine adenovirus strain 6618 infection were studied in 18 hysterotomy-derived, colostrum-deprived piglets, by histology, transmission electron microscopy, and an immunoperoxidase technique. Viral particles could be seen in altered epithelial cells of the lower small intestine from 24 hours until 16 days after inoculation. With the immunoperoxidase technique, viral antigen could be found in epithelial cells at 45 days after challenge. Destruction of epithelial cells and shortening of villi could be related directly to the presence of the virus. Histology of the lower jejunum and ileum, which demonstrates intranuclear inclusion bodies, is proposed as a useful diagnostic technique. PMID- 6280364 TI - Ultrastructural changes in the small intestine of neonatal calves with enteric colibacillosis. AB - The small intestines of calves inoculated orally with the enteropathogenic strain of Escherichia coli 0101:K'B41',K99 were examined by electron microscopy at 3, 6, 12, 16, 21, 36, 69, 70 and 72 hours after inoculation. The challenge organism adhered to the mucosa of the distal small intestine from six hours post inoculation. Bacteria were separated from the microvillous brush border by a gap of 200 to 300 nm in which bacterial fimbriae and the microvillous glycocalyx were seen. Bacteria never were found in epithelial cells but were present in macrophages in the lamina propria from 12 hours. At three and six hours, cytopathic changes were not seen in the small intestine, but from 12 hours epithelial cells on affected villi had blunt and thick microvilli and contained cytoplasmic inclusions. Epithelial cells were seen frequently in the process of extrusion from the villi, either singly, in small groups, or as ribbons of cells. Intervillous bridges, characteristic of villous fusion, were seen frequently from 69 hours. PMID- 6280366 TI - An outbreak of paresis in mares and geldings associated with equid herpesvirus 1. AB - An outbreak of paresis occurred on a small isolated stud farm in July 1980. Of the 42 horses on the stud, infection was confined to a group of nine in-foal mares and their foals and eight other horses which were either housed together at night or grazed adjacent pastures. Eight mares and two geldings developed ataxia or paresis and one mare died. Equid herpesvirus 1 was isolated from 17 animals and serological studies confirmed that 24 of 26 animals sampled had experienced infection. PMID- 6280365 TI - Ultrastructural evaluation of the acute encephalitis and hydrocephalus in dogs caused by canine parainfluenza virus. AB - Intracranial infection of neonatal dogs with canine parainfluenza virus (strain 78-238) progressed from an acute phase of encephalitis to a chronic phase of internal hydrocephalus. Acute encephalitis was characterized ultrastructurally by laminar cortical necrosis, neuronal degeneration, neocapillary formation, reactive gliosis, and ependymitis. Viral nucleocapsids were seen in the cytoplasm of neurons. Ultrastructural lesions in hydrocephalic dogs were restricted to ventricular surfaces. The earliest light microscopic change in ependymal cells occurred four weeks post-infection and consisted of segmental loss of ependymal cells. Dogs killed at three and six months post-infection had quantitatively similar ultrastructural alterations in ependymal cells regardless of the extent and severity of hydrocephalus. The shape of these cells changed from cuboidal to squamous. There was progressive loss of cilia and cell organelles along with a concomitant increase of cytoplasmic filaments. Viral nucleocapsids and inflammatory cells were not present in convalescent hydrocephalic dogs. These results suggested that virus-associated hydrocephalus can occur in the absence of obvious viral persistence in ependymal cells and in the absence of inflammation in the ventricular system. PMID- 6280367 TI - Immune responses of sheep to louping-ill virus vaccine. AB - The immune responses of sheep to single and double doses of commercially available louping-ill virus vaccine were examined. The susceptibility to challenge of sheep which had been vaccinated but showed a poor response was also investigated. Two injections of vaccine were required to provoke an adequate antibody response and maximum titres were obtained when there was an interval of two to eight weeks between injections. After challenge, viraemia could not be detected in animals with an antibody titre of 20 although increase in the concentration of humoral antibodies indicated that infection had occurred. Vaccinated but seronegative sheep and vaccinated animals with an antibody titre of 10 were also clinically resistant to the challenge, although circulation of virus was demonstrated. That vaccination had sensitised those animals to viral antigen was evident from the reduced viraemias, the early rise in humoral antibody titres and subsequent protection afforded compared to unvaccinated control animals. Thus, animals with minimal antibody titres after vaccination are protected, but it is recommended that vaccines eliciting the highest possible antibody responses will be the most useful under field conditions. PMID- 6280368 TI - Enzyme-linked immunosorbent assay for detecting antibodies to Aujeszky's disease virus in pigs. AB - An indirect micro enzyme-linked immunosorbent assay (ELISA) for detecting antibodies to Aujeszky's disease virus in pigs is described. A control antigen prepared from infected cells was included for each serum tested. Of 243 sera from serologically positive farms, 175 (72 per cent) and 147 (60 per cent) were positive by the ELISA test and microtitre serum neutralisation test, respectively. Failure to include a control antigen for each serum would have resulted in 14 sera (6 per cent) being differently recorded. Results for sera from experimental and field infections indicated that seroconversion was more quickly detected by the ELISA test than the microtitre serum neutralisation test. In addition to greater sensitivity the ELISA test has other advantages over the serum neutralisation test. ELISA is a rapid, cheap test which is not dependent on a continuous supply of cell cultures and which can be readily automated. PMID- 6280369 TI - Use of control antigen to improve the enzyme-linked immunosorbent assay for enzootic bovine leukosis antibodies. PMID- 6280370 TI - Immunity to canine adenovirus respiratory disease: a comparison of attenuated CAV 1 and CAV-2 vaccines. AB - Four litters of puppies were divided into three groups. One group was vaccinated with a live CAV-1 vaccine and another with a live CAV-2 vaccine. Throat swabs were collected from two dogs in each of these groups to monitor the possible excretion of vaccine virus, but none was found. Both groups, together with the third group of unvaccinated controls, were challenged 17 days later with an aerosol of virulent CAV-2. One dog from each group was killed on the third, fourth, seventh, ninth, 11th and 14th days after challenge. The unvaccinated dogs developed a clinical disease characterised by anorexia, dullness, coughing and tachypnoea. The lungs were consolidated and histological examination revealed the main lesion to be a severe necrotising bronchiolitis. Large amounts of virus were present in the respiratory tissues of these dogs and high titres of virus were isolated from throat swabs. In contrast, both groups of vaccinated dogs remained clinically almost normal with minimal lesions, present for a much shorter period of time. Virus was found on day 4 in the respiratory tissues of one dog vaccinated with CAV-1 but the other vaccinated animals contained little or no virus. In general, the degree of protection afforded by CAV-1 vaccine seemed similar to that provided by CAV-2 vaccine. PMID- 6280371 TI - [Synostosis of hand and foot bones]. PMID- 6280372 TI - Pituitary adenomas with hyperfunction of TSH. Frequency, histological classification, immunocytochemistry and ultrastructure. AB - In a collection of 564 surgically removed pituitary adenomas, 4 cases werefound to have had elevated TSH plasma levels. One of these tumors )(case 1) could be classified as a highly differentiated mucoid TSH cell adenoma presenting histochemical reactions typical of, as well as electron microscopical features identical to, normal TSH cells. Immunoenzymatic studies failed to dfemonstrate TSH in the tumor cells. Two further adenomas (case 2 and 3) were similarly structured in many areas, but showed regions of poorer differentiation in which distinct pleomorphism, irregular secretory granules, increased numbers of ribosomesand a well developed rough endoplasmic reticulum were present. In 10% of the tumor cells GH could be demonstrated immunoenzymatically, but there was no TSH. The fourth adenoma was an undifferentiated acidophilic adenoma showing pleomorphic cells having slight acidophil and partly mucoid granulations. The ultrastructure showed convoluted nuclei, increased numbers of free ribosomes as well as abundant rough endoplasmic reticulum and secretory granules which were differentin size and number but distinctly of the TSH cell type. Immunoenzymatically, TSH was found in some cells, with GH in more cells. Endocrinologically, elevated levels of GH were measured in cases 2, 3 and 4 with LH being increased in case 1. Clinical and morphological correlations are discussed. PMID- 6280373 TI - Results of an etiological therapy applied to patients with different localizations of herpes infection detected by immunofluorescence reaction. AB - An indirect immunofluorescence reaction was used for cytovirological diagnosis in 110 patients with different herpes-like clinical manifestations. The presence of herpes antigens type 1 and 2 was made evident in the exfoliated cells from 77% of the cases with recurrent herpes, in 68% of those with cutaneous and/or oral herpes, in 64% of the cases with keratoconjunctivitis and in 37.5% of the women with different genital diseases. Administration of specific antiherpes immunoglobulins and of the chemotherapeutic drug Flumidin to cases with positive immunofluorescence reactions led to improvement of the symptoms and even to clinical recovery. PMID- 6280374 TI - Some characteristics of Sendai virus envelope solubilization by different detergents. AB - Gel-filtration through Sepharose-2B of Sendai virus envelopes solubilized by Tween-20 or Triton X-100 treatment allows the separation of two elution peaks, hemagglutinin (HA) activity being detected only in the peak corresponding to large envelope fragments. Solubilization of the same virus envelopes with N octylglycoside (NOG) results in a single elution peak, exhibiting both HA and neuraminidase activities. The results obtained suggest : 1) the similar envelope composition of the different virus populations, and 2) the different mechanisms of solubilization, in relation to the detergent used for disruption. The advantages of virus disruption by NOG are discussed. PMID- 6280375 TI - Relationships between ceruloplasmin and viral immunity. AB - Experimental data are reviewed with reference to the role of ceruloplasmin as a nonspecific factor of viral immunity. Ceruloplasmin would act a) by an inhibition of virus multiplication, resulting in a "slow-motion" infection, which promotes immunization; b) by leading to the synthesis of a virus population rich in defective interfering particles, and c) by the direct modulation of the immune response. PMID- 6280376 TI - Phosphonoacetic acid effective in herpesvirus infection. PMID- 6280377 TI - Investigations in the transforming activity of temperature-sensitive mutants of herpes simplex virus. AB - Transformation of rat embryo fibroblast was induced by a temperature-sensitive mutant of herpes simplex virus type 2. Transformation was scored using the morphological criterion of focus formation and the cell changes that lead to the final focus are described. Virus genome persistence in the transformed cell line was demonstrated by the presence of virus-specific membrane antigens and by the specificity of antibodies elicited in rabbits by inoculation of transformed cells. The cytogenetic analysis of the transformed cells revealed changes in chromosome number and the presence of marker chromosomes. PMID- 6280378 TI - An SV40 deletion mutant accumulates late transcripts in a paranuclear extract. PMID- 6280379 TI - Viable polyoma virus variant with two origins of DNA replication. PMID- 6280380 TI - Oligosaccharide modifications and the site of processing of gPr92env, the precursor for the viral glycoproteins of Rous sarcoma virus. PMID- 6280381 TI - Intracellular transport of SV40 large tumor antigen: a mutation which abolishes migration to the nucleus does not prevent association with the cell surface. PMID- 6280382 TI - Inhibitors of glycosylation reverse retroviral interference. PMID- 6280383 TI - DNase I sensitivity of endogenous and exogenous proviral genome copies in M-MuLV induced tumors of Mov-3 Mice. PMID- 6280384 TI - Virus-specific transcription in bovine papillomavirus-transformed mouse cells. PMID- 6280385 TI - Circular monomers of bacteriophage lambda DNA as substrates for in vitro packaging. PMID- 6280386 TI - XVIII. Effective treatment of AKR leukemia with antibody to gp7 1 eliminates the neonatal burst of ecotropic AKR virus producing cells. PMID- 6280387 TI - High multiplicities of infection favor rapid and random evolution of vesicular stomatitis virus. PMID- 6280388 TI - [Benzo(a)pyrene content in sausage products]. AB - The content of the carcinogenic polycyclic aromatic hydrocarbon, benz(a)pyrene was determined in sausage products (boiled, raw- and semi-smoked) treated both with a smoke-air mixture and with the smoking liquid "Vakhtol". It was found that in raw- and semi-smoked sausage products treated with the smoke-air mixture, the level of benz(a)pyrene was 12-18 times greater than in boiled sausages exposed to the similar treatment. The use of the smoking liquid "Vakhtol" favours the manufacture of the produce not contaminated with carcinogenic substances. This forms basis for "Vakhtol" introduction into food industry. PMID- 6280389 TI - [Hormone-dependent process of the maturation of the major structural proteins of the mouse mammary tumor virus]. AB - The following findings were obtained by the radio-immunoprecipitation method with antisera to gp52 and p27. When the cells were cultivated in a hormone-free medium, they contained precursors of proteins of gene gag (Pr 73gag) and gene evn (gPr 70env). No mature structural MTV proteins were found. The addition of insulin to the growth medium had no effect on maturation of protein precursors. When dexamethazone was added to the medium, gPr 70env "maturated" to gp52 of the main envelope glycoprotein of virion coat whereas Pr 73gag was not cleaved into final products. When the cells were cultivated in the presence of insulin and dexamethazone, there was a marked stimulation of Pr 73gag (although its maturation did not occur), g Pr 70env, and, especially, gp 52. Extracellular particles were found to contain p27 which was precipitated by homologous monospecific antiserum, that is, in the system of clone F2 cells processing of Pr 73gag occurred mainly in extracellular particles. Thus, expression of all three known genes (gag, pol, and env) occurred in the cells of cloned F2 culture. PMID- 6280390 TI - [Formation of transfecting DNA in acute cellular infection with the tick-borne encephalitis virus mediated by an oncogenic virus]. AB - To prove the association of the transfecting activity of nucleic acid from cells co-infected with TBE and SV40 viruses with DNA, experiments were carried out using fractionation of nucleic acid preparations in cesium sulphate gradient an on a column with HAP. The experiments led to a conclusion that the infectivity of the preparations used was associated with double-stranded DNA. Investigation of nucleic acid infectivity form cytochalasine-enucleated cells indicated that the function of the nucleus was necessary for formation of transfecting DNA in mixed TBE and SV40 infection of cell. No formation of transfecting DNA was observed in experiments of blocking replication and transcription of cellular genome. The retention of the transfecting activity of DNA during 6 passages of cells at approximately the same level indicated the lack of selective preferences or inhibition of cells containing transfecting DNA. PMID- 6280391 TI - [Karyological characteristics of L cells and the identification with them, of the antigen of persisting Japanese encephalitis virus]. AB - The immunofluorescence procedure revealed the virus-specific Japanese encephalitis virus antigen in latently infected L cells as well as differences in the percentage of antigen-positive cells and in the localization of the virus specific antigen under conditions of acute, chromic, and latent infection and in the period of activation of the latent state of the virus at 28 degrees C. Karyological studied demonstrated no marked differences between persistently infected and original lines. PMID- 6280392 TI - [Isolation of a cytopathogenic rotavirus from calves]. AB - A cytopathic rotavirus was isolated from calves with symptoms of diarrhea and underwent 15 passages in bovine embryo kidney cell culture. The specificity of the isolate was confirmed by the neutralization test, immunofluorescence, electron microscopy. The CPE of the isolated in bovine embryo kidney cells was similar to that of the reference strain. PMID- 6280393 TI - [Morphological and structural studies of Sendai virus ribonucleoprotein]. AB - The structure and morphology of Sendai virus ribonucleoprotein (RNP) with native and split protein subunits were studied by the method of circular dichroism and electron microscopy. Cleavage of the polypeptide fragment from RNP protein subunits was shown to cause changes in the secondary structure of RNP in situ but not to affect the RNP morphology; when stained with uranyl acetate both RNP types had an appearance of spiral strands. PMID- 6280394 TI - [Effect of formaldehyde on the ribonucleoprotein structure of Sendai virus]. AB - The effect of formaldehyde on the conformation of Sendai virus ribonucleoprotein (RNP) was studied by electron microscopy and a spectrophotometric method. The effect of formaldehyde on RNP conformation was found to depend strongly on the ionic strength of the solution. Under conditions of a low ionic strength in the presence of 1.5% formaldehyde the tightly coiled rods of RNP stained with uranyl acetate become loosely coiled or almost completely extended. At the same time, formaldehyde reaction with RNP results in a decrease in extinction of the RNP suspension in the 250 to 290 spectral region. However, RNP incubation with formaldehyde in the presence of 0.5M NaCl caused no changes in the morphology and spectral properties of the RNP under study. The results indicate that formaldehyde cannot be used for fixation of Sendai virus nucleocapsids before negative staining for electron microscopy examinations. PMID- 6280395 TI - [Physicochemical properties of Teschen disease virus RNA]. AB - The specific infectivity of virion RNA of teschen disease virus in a sensitive PP cell culture was 4-5 lg TCD50/ml per 1 microgram RNA. When virion RNA was inoculated into cell cultures insusceptible to the native virus, the virus replicated to a titre of 2.0-3.5 lg TCD50/ml. The molecular weight of virion RNA determined by two independent methods was 2.7 x 10(6) daltons. Tm calculated from the curve of virion RNA melting temperature was 57 degrees C. The double-stranded replicative form of RNA recovered from virus-infected PP cells was shown to have sucrose gradient sedimentation coefficient of 20 S. The specific infectivity was 2-3 lg TCD50/ml per 1 microgram of RNA. PMID- 6280396 TI - [Cathepsin D activity in cells differing in their sensitivity to the poliomyelitis virus]. PMID- 6280397 TI - [Effect of riodoxol on herpes simplex virus morphogenesis in experimental eye infection]. AB - The effect of riodoxol on herpes simplex type 1 viruses was studied. It was shown to affect multiplication and maturation of the virus as manifested in the appearance of virions with incomplete nucleoid structure and long-term presence of virions in cytoplasmic vacuoles in cells. Submicroscopic changes suggest a dual effect of riodoxol on herpes simplex virus: on the DNA replicative cycle and on the stage of late maturation of virus at the time of its release from the nucleus.' PMID- 6280398 TI - Nutrition cultism. PMID- 6280399 TI - Tumor angiogenesis factor with possible implications for glioblastoma multiforme. PMID- 6280400 TI - [Social aspects in fighting against tuberculosis]. AB - In 1884 Robert Koch postulated in his essay on "The Etiology of Tuberculosis" favouring moments for the settling of tubercle bacilli to explain the predisposition and diversified course of tuberculosis in grown-up persons. Under the conception "disposition" he also subsumised exogen factors beside the internal conditions and proclaimed that all steps undertaken against tuberculosis should take into account the social conditions. Only to effect the treatment in a sanatorium for such patients, it is also necessary to care for their families and for the patients themselves after they left the sanatorium. Complex fighting programmes developed according to the thorough analysis of the social surroundings, including the influences of occupation, income, expenses for food, housing and clothing, the legislation with regard to the health law and education to health duty as well as the level of medical science. Briefly, there was stated that the tuberculosis mortality depends on the standard of life. From the war epidemiology of tuberculosis at least 3 parallel curves resulted, decisively influencing the mortality, the curves of infection and charging with work as well as the curve of nourishment. So the tuberculosis was characterised as a social disease. No wonder that social appearances--and war is one of them--stamp the special picture. As a model of applied knowledge, the strategy of fighting tuberculosis in the GDR and its outstanding results are discussed. After the antituberculosis chemotherapy was introduced, which shortens the duration of the disease and removes its chronic relapsing course, the significance of the social factors, however, decreased, but concerning the situation in the Third World, their knowledge is of great importance and not only of historical interest. PMID- 6280401 TI - [Production of hyper-immune serum for diagnosis of herpes virus hominis infections]. PMID- 6280402 TI - [Passive hemagglutination for the detection of humoral antibodies against rhinoviruses]. PMID- 6280403 TI - [Pheochromocytoma and shock]. AB - In phaechromocytomas with predominant beta-adrenergic activity in a short time develop hypovolaemia and shock due to capillary transudation and profuse sweating. On the basis of an own observation the following symptoms may be summarized: signs of centralisation such as cold, pale, sweat-covered skin, cool extremities in increased nuclear temperature, clear decrease of the blood pressure, tachycardia, arrhythmia, signs of the left heart insufficiency and considerable changes of the ECG as an expression of a catecholamine-induced cardiomyopathy, strong abdominal pains, strong feeling of thirst in isotonic dehydration, oliguria and metabolic acidosis. In patients with shock syndrome of unclear etiology should also be thought of a phaeochromocytoma after exclusion of the most frequent causes. PMID- 6280404 TI - [The value of surgical studies for the current state of diagnosis and treatment of bronchial cancer]. AB - A critical consideration of the modern situation of the diagnostics and therapy of the bronchial carcinoma shows compared with the expense a proportionately small healing quotient of nearly 7% of all diagnosed bronchial cancers. The analysis of the possibilities of the present ways for the therapy shows that an additional improvement of the operative and immediate postoperative results could not lead to a substantial modification of the healing quotient altogether. An improvement is possible only through an increase of the number of operable patients at the moment of the diagnosis. This requires a radical modification of the methods for the early diagnosis. Possible ways for them are shown. A smaller improvement of the healing quotient could be achieved by the intensification and the correction of the after treatment in the later time, inclusively the well timed and effective treatment of relapse and metastases. The healing quotient after five years may probably be doubled with the right use of the above mentioned measures. PMID- 6280405 TI - [Erythema multiforme as a vesicular exanthema (author's transl)]. AB - We report about a special case of recurring Erythema multiforme after Herpes glutealis. There closely grouped vesicles all over; only at the palms we saw target-like skin lesions. PMID- 6280406 TI - [Macrophage secretion]. PMID- 6280407 TI - [Pseudosarcoma Kaposi's: clinical manifestations and relationship with congenital vascular dysplasias]. PMID- 6280408 TI - Identification of neoplasia, especially hepatoma, as a major cause of mortality in the Southern Highlands. AB - This study shows that relative to the number of deaths in hospital, the number of patients discharged home to die is high, that the majority of these cases have a malignancy and that the prognosis of these 'home to die' (HTD) patients is accurate. Respiratory disease is the leading cause of death overall in the community no matter what data are included. The inclusion of HTD cases in hospital mortality data reveals that malignant disease is the leading cause of death in adults and the second cause of death overall and that hepatoma is the major malignant disease. The relevance of a hepatitis B vaccination programme for P.N.G. is discussed. PMID- 6280409 TI - [Simultaneous representation of EsD, DIA, GLO, and UDPGP in a starch block (author's transl)]. PMID- 6280411 TI - [A possible misinterpretation of dead bone decomposition features by macroscopic inspection (author's transl)]. PMID- 6280410 TI - [A new allele of AK-Polymorphism: AK0 (author's transl)]. PMID- 6280412 TI - In vitro susceptibility of salmonellae to eight antimicrobial agents. AB - The Bauer-Kirby disk diffusion method was used to determine the susceptibility of 333 strains of Salmonella isolated in Saudi Arabia to eight antimicrobial agents. The following susceptibilities were observed: Cephalothin 99.40%, furadoine 99.10%, chloramphenicol 98.80%, ampicillin 98.20%, polymyxin 96.70%, kanamycin 95.80%, streptomycin 90.99%. Only 50.45% of the strains were susceptible to tetracycline, whereas 44.14% and 5.41% were respectively intermediate and resistant to this drug. The antibiograms displayed many resistant patterns. Some of the salmonellae were singly resistant while others were resistant to 2 or more drugs in various combinations. In view of the potential for salmonellae to have inherent or acquired resistance to certain commonly used antimicrobial agents, susceptibility of all isolates of salmonellae, particularly of all clinically significant strains should be determined periodically. PMID- 6280413 TI - [A contribution to the recovery of poliomyelitisvirus from digested sewage sludge dependent on pH-value (author's transl)]. AB - For later investigations on the influence on virus in digested municipal sewage sludge four different methods were tested to recover virus of poliomyelitis from the sludge. The best results were obtained by elution with 3% beef-extract at pH 9, followed by ultrasonic-treatment. The recovery-rate is not significantly influenced by the final concentration of beef-extract. Poliovirus inoculated to anaerobic digested sludge shows a 90% reduction within 7 hours at room temperature. PMID- 6280414 TI - [Removal of polio- and parvovirus in sewage-sludge by lime-treatment (author's transl)]. AB - Lime (CaO) applied as conditioner for dewatering municipal sewage sludge in filter-presses is suitable to disinfect sewage sludge with respect to virus. A pH rise to more than 12 followed by release of ammonia inactivates Polio- and Bovine Parvovirus. Addition of 3 kg CaO and more per m3 of raw sludge inactivates Poliovirus within 30 minutes. 7.5 kg CaO per m3 of anaerobically digested sludge are required to inactivate Polio-virus within 5 days and Bovine Parvovirus within 24 hours. Parvovirus seems to be more sensitive against free ammonia than Poliovirus, but less sensitive against high pH-values. PMID- 6280415 TI - [Study of the microbial multiplication in the muscle just after slaughter (author's transl)]. AB - The bactericidal or bacteriostatic activity of the serum, the muscle and the rabbit meat juice was followed after slaughter on animals vaccinated or not against E. coli, P. putida, Cl. perfringens. Rabbit serum is bactericidal against gram negative bacteria, P. putida and E. coli. On the contrary it is without effect on Cl. perfringens. Muscle has no inhibitory effect against E. coli and Cl. perfringens on animals vaccinated or not. P. putida is inhibited or partially eliminated from rabbit muscle just after slaughter. Vaccination does not increase the bacteriostatic or bactericidal activities of the sera, the meat juices and intact muscles. PMID- 6280416 TI - [Electromyography and electroneurography in peripheral nerve lesions. Part II]. AB - Electromyographical and electroneurographical examinations are indicated in the forefield of the surgical treatment for radicular syndromes, traumatic plexus and nerve lesions as well as compression or bottle-neck syndromes. The electrophysiological findings point out the neurogenic nature of the disturbance. They furthermore furnish statements with respect to the localisation, the extent, the kind and the prognosis of the lesions. The electromyographical and electroneurographical examination programme is determined by the anatomical innervation conditions and the questions that are of clinical interest. In case of radicular syndromes it is mainly the localisation with respect to the height of the root lesions that is of interest. In plexus lesions, the question regarding the reinnervation chance is usually to the fore. In primary nerve lesions, the question whether there is a partial or total nerve separation is above all of interest. In secondary nerve lesions and the bottle-neck syndromes it is usually necessary to determine the exact site of the lesions and the extent of the chronic compression damage. In this paper, these aspects ae discussed on the basis of the more important clinical pictures and practical hints for diagnosis and therapy are given. PMID- 6280417 TI - [Anatomic pathways of pain irradiation in lumbar osteochondrosis]. PMID- 6280418 TI - [Features of encephalitis developing against a background of acute respiratory diseases]. PMID- 6280419 TI - [Relation between mental and endocrine disorders: psychoneuroendocrinology of affective disorders]. PMID- 6280420 TI - Isolation and characterization of phosphatases from vascular smooth muscle. AB - Actomyosin preparations of the carotid arteries of cattle contain a soluble phosphatase activity, which can be removed from the contractile proteins by repeated washings. This enzyme activity is lowered by high ionic strength, potassium fluoride, zinc acetate, ammonium molybdate, and vanadate. Magnesium ions enhance the enzyme activity. The phosphatase activity shows a maximum between pH 5.5 and 6.0 and a plateau of pH 7-9. By means of gel filtration on Sepharose 6B the phosphate activity is separated into three peaks, which are characterized with respect to their inhibition by potassium fluoride, ammonium molybate, and vanadate and their dependence on pH. PMID- 6280421 TI - A specific method of high sensitivity for the determination of adenosine in the incubation medium of fat tissue. AB - A suitable method for the measurement of adenosine in the incubation medium of fat tissue (200-500 mg) has been developed. The method is based on the specificity of the adenosine deaminase reaction and on the high sensitivity of a fluorescent method for adenine derivatives. The decrease of fluorescence in a sample after treatment with this enzyme is used for measuring adenosine in the range of 50-500 pmoles/tube. This method is highly specific and is not affected by other adenine derivatives present in the sample. Instead of acetic acid, perchloric acid was used in the fluorescent reaction, thus increasing the amount of adenosine dependent fluorescence. With this modification of the original fluorescent method, perchloric acid extracts can be used without further processing after deproteinization of the samples. Using this method, we could measure the adenosine release of fat pads of Wistar rats incubated in Krebs Ringer-albumin buffer without concentration or purification procedures. PMID- 6280422 TI - [Intra-thyroid metastases (author's transl)]. AB - Four cases with intra-thyroid metastases which simulate a primary tumor of the gland are reported. These metastatic lesions are probably more frequent than generally accepted. Their clinical discovery is quite frequent in patients with a renal neoplasm and is in those cases often the presenting feature of the primary tumor. Ablation of the thyroid gland is often necessary to make a correct diagnosis and to prevent local compression symptoms. This intervention is indicated whenever deemed necessary unilateral thyroid lobectomy is preferable to total ablation since it achieves its goal with a lower mortality. Postoperative radio- and chemotherapy might be added as indicated. PMID- 6280424 TI - Accelerative effect of adenosine 3', 5'-cyclic monophosphate and dibutyryl adenosine 3', 5'-cyclic monophosphate on the cleavage cycle of the sea urchin eggs. PMID- 6280423 TI - Clinical aspects of GIP secretion. AB - The gastric inhibitory polypeptide (GIP) is the main hormone of the incretin type acting on the entero-insular axis. It is released after fat, glucose or meal ingestion. The variations of this secretion are described in obesity and in some pancreatic and gastrointestinal diseases: it is increased in maturity onset diabetes mellitus, obesity or duodenal ulcer, variable according to the food taken and the severity of the pancreatic lesion in chronic pancreatitis and cystic fibrosis, normal in insulinoma and decreased in celiac disease. The impaired absorption of the food-stuffs and the defective feed-back regulation of GIP secretion by insulin are the major causes of these variations. To a lesser degree, gastric acid secretion, gastric emptying and vagal control may also influence GIP secretion. PMID- 6280425 TI - Acute suppressive effect of ACTH-induced cortisol secretion on serum prolactin levels in healthy man. AB - The effect of ACTH on serum prolactin concentrations was studied in 6 healthy women in the follicular phase of the menstrual cycle, in 5 healthy men and in 6 patients (5 males, 1 female) with adrenocortical insufficiency. In healthy women prolactin levels decreased from basal, 14.4 +/- 2.1 (SEM) microgram/l to 9.4 +/- 0.9 microgram/l after 30 min and to 8.1 +/- 0.7 micrograms/l after 60 min of iv administration of synthetic ACTH1-24 (0.25 mg). Upon continuous infusion of ACTH1 24 (0.25 mg for 8 h) prolactin fell to 4.4 +/- 0.6 microgram/l in healthy women and to 4.6 +/- 1.5 micrograms/l (basal: 10.6 +/- 1.8 micrograms/l) in healthy men. In patients with adrenocortical insufficiency prolactin concentrations remained unchanged following an 8 h infusion of ACTH1-24 (before ACTH: 14.5 +/- 2.3 micrograms/l, after ACTH: 16.3 +/- 3.1 micrograms/l). After treatment with dexamethasone (2 mg/day for 3 days) however, prolactin concentrations were suppressed both in healthy women (-52 +/- 7%) and men (-25 +/- 11%) and in patients with adrenocortical insufficiency (-21 +/- 10%). Thus the effect of ACTH on prolactin appeared to be mediated via enhanced cortisol secretion. It is suggested that an acute increase in cortisol levels within the physiological range may modulate prolactin secretion. PMID- 6280426 TI - ACTH-producing pituitary adenomas in Addison's disease: two cases treated by transsphenoidal microsurgery. AB - In 2 women with known Addison's disease, progressive hyperpigmentation reappeared years after an initial remission under conventional substitution therapy with cortisone. Excessively elevated plasma ACTH concentrations and radiological evidence of sella turcica deformation led to the diagnosis of ACTH-producing adenomas and prompted their removal by transsphenoidal microsurgery. In one patient, a large Crooke's cell adenoma with extensive extrasellar expansion had caused severe and irreversible bilateral defects of the visual fields and unilateral optic atrophy. Surgical removal of the tumour and radiotherapy brought about a permanent disappearance of the hyperpigmentation, but eventually led to secondary hypothyroidism. In the second patients, the selective removal of a small intrasellar eosinophilic adenoma consisting of ACTH-producing cells did not alleviate the hyperpigmentation and did not lower the plasma ACTH concentration. However, hyperpigmentation regressed markedly within a year of treatment with a higher dose of cortisone. The rarity of similar cases in the literature seems to indicate that insufficient feedback suppression of ACTH-producing cells in treated Addison's disease does not by itself induce the development of a pituitary adenoma, but might promote the growth of an independently and coincidentally occurring microadenoma, which would have caused Cushing's disease in a person with intact adrenal glands. PMID- 6280427 TI - A case of ectopic ACTH syndrome: diagnostic difficulties caused by intermittent hormone secretion. AB - A patient with a thymic carcinoid tumour causing ectopic ACTH syndrome is presented. The case illustrates the rapid development of the clinical and laboratory findings often associated with ectopic secretion of ACTH, including severe proximal myopathy, emotional lability, and hypokalaemic alkalosis. Interpretation of conventional tests of pituitary-adrenal function was complicated by intermittent secretion of ACTH by the tumour. The results of selective venous sampling for ACTH ruled out pituitary ACTH hypersecretion and were suggestive of a thymic source; computerized tomography of the chest localized the tumour. In vivo and in vitro investigations confirmed excessive ACTH production by the tumour, and surgical resection plus radiotherapy has resulted in resolution of the syndrome. The diagnostic problems created by intermittent secretion of ACTH by these tumours and the pre-operative and post operative medical management of these patients are discussed. PMID- 6280428 TI - Factors influencing the release of cyclic AMP from mouse thyroid tissue stimulated by TSH in vitro. AB - The accumulation of cyclic AMP (cAMP) in mouse thyroid tissue in response to TSH in the presence of 1 mM theophylline was accompanied by a release of the nucleotide from the tissue into the incubation medium. This cAMP release was almost rectilinearly related to the time of exposure to TSH, and rectilinearly related to the log concentration of TSH in the range 0.1-5 mU/ml. The cAMP release proved to be independent of the pre-incubation time up to 4 h, and took place also in the absence of methylxanthines when the cAMP level was low. The total cAMP accumulation in response to TSH was augmented by different inhibitors of protein synthesis but the fraction of the nucleotide that was retained intracellularly was increased only by puromycin. Dipyridamole had an effect similar to that of puromycin. Depolarization or treatment with ouabain did not change the distribution of cAMP between tissue and medium. It is concluded that the release of cAMP from thyroid tissue stimulated by TSH may take place under physiological conditions, that it seems to be regulated by the actual concentration of TSH, and that it may be of significance for the regulation of the intracellular cAMP level. PMID- 6280429 TI - Development of the gonadotrophic resistant ovary syndrome in myasthenia gravis: suggestion of similar autoimmune mechanisms. AB - A woman with myasthenia gravis who developed hypergonadotrophic amenorrhoea was studied. This patient fulfilled all accepted criteria for the diagnosis of the gonadotrophin resistant ovary syndrome: high levels of serum LH and FSH by radioimmunoassay and urinary gonadotrophin excretion by bioassay, low serum oestradiol, lack of response to exogenous gonadotrophin and ovaries with multiple non-stimulated primordial follicles. The serum of this patient contained a substance which behaving like a gamma globulin, inhibited FSH specific binding to receptors in an in vitro system. This inhibitor was looked for in further patients with hypergonadotrophic amenorrhoea, other immunological diseases or myasthenia gravis with normal menses. Ovarian biopsy was not available in these patients. In all except one who also had the association of myasthenia gravis and hypergonadotrophic amenorrhoea, the inhibitor was not detected. It is suggested that development of antibodies directed against gonadotrophin receptors or a receptor related membrane domain may play a role in the mechanism of failure of gonadotrophin stimulation in some patients with the gonadotrophin resistant ovary syndrome. PMID- 6280430 TI - Observations on the effects of prolactin on LH-receptors and steroidogenesis in corpus luteum and testis of the hypophysectomized rat. PMID- 6280431 TI - Effects of antibodies to catecholoestrogens and catecholoestrogen methyl ethers on PMS induced ovulations in immature rats. PMID- 6280432 TI - Properties of thyrotrophin receptors in human thyroid membranes. AB - The influence of pH, salts, temperature and time of incubation on the binding of [125I]thyrotrophin to its human thyroid receptor was studied. Optimal binding was achieved at pH 7.0--7.6. Pre-incubation of membrane receptors at pH 6.0 resulted in a 60% loss of binding. There was dynamic equilibrium, as demonstrated by kinetic experiments. Scatchard analysis yielded non-linear plots with apparent affinity constants of 1.9 x 10(9) M-1 and 3.7 x 10(7) M-1. The concentration of [125I]thyrotrophin used in the experiments (1.2--2.2 pM) approximated normal human serum thyrotrophin concentration. PMID- 6280433 TI - Refractoriness of TSH- and PGE2-stimulated iodine metabolism in cultured porcine thyroid cells, evidence for refractoriness at the level of cAMP action. AB - In an attempt to study intrinsic regulatory mechanism involved in iodine metabolism, chronic and acute effects of TSH, PGE2 and DBC on iodine uptake, iodide discharge and organic binding of iodine were examined using cultured porcine thyroid cells. Culture in the presence of TSH, PGE2 and DBC for 6 days maintained the ability to thyroid cells to take up iodine and organify it, but culture in the absence of these substances failed to do so. When incubated with NaI in the presence of 1 mM methylmercaptoimidazole (MMI), the cells took up iodide and this accumulated iodide was discharged by TSH, pGE2 and DBC. TSH-, PGE2, and DBC-stimulated iodide discharge was depressed greatly after chronic exposure to TSH, PGE2 or DBC. This refractoriness of TSH-, PGE2- or DBC stimulated iodide discharge was not specific for each thyroid stimulating substance; previous exposure to TSH, PGE2 or DBC induced refractoriness of TSH-, PGE2- and DBC-stimulated iodide discharge, providing evidence for the existence of refractoriness at the level of cyclic AMP action on iodide discharge. When incubated with NaI in the absence of MMI, the cells took up iodide and organified it. After 30 min incubation with NaI, TSH, PGE2 and DBC were added and they stimulated iodide organification further. This TSH- and PGE2-stimulated iodide organification was also depressed after exposure to TSH or PGE2. These data indicate that, as an intrinsic regulatory mechanism, refractoriness is operating at the level of cAMP action on iodine discharge and organification. PMID- 6280434 TI - Inhibition by calcium channel blockers of the glycogenolytic effect of glucagon in perfused rat liver. AB - Verapamil and diltiazem, calcium channel blockers, inhibited significantly the glucagon-induced glucose output and 45Ca efflux from perfused rat liver at concentrations higher than 50 microM when the perfusate contained calcium. Although the blockers partially interfered with glucagon-induced elevation of cyclic AMP in the tissue, they also inhibited the effects of cyclic AMP. The blockers did not show the inhibitory effects in the absence of perfusate calcium. However, the inhibition of calcium influx into hepatocytes by omission of extracellular calcium or addition of EGTA did not interfere with these effects of glucagon and cyclic AMP. In the presence of extracellular calcium, the blockers did not inhibit cyanide-induced glucose output, indicating that the activity of glycogen phosphorylase and later processes leading to glucose output were not affected by the blockers. These data suggest that, in the presence of calcium, the blockers inhibit the effect of glucagon also at a step (or steps) subsequent to cyclic AMP production and before the activation of phosphorylase b, probably by inhibiting glucagon-induced mobilization of calcium from intracellular calcium pools rather than inhibiting calcium influx into hepatocytes. PMID- 6280435 TI - Isolated ACTH deficiency and primary hypothyroidism. AB - A patient presented with severe primary hypothyroidism and secondary adrenal insufficiency due to the isolated deficiency of ACTH. The diagnostic evidence suggests that both are of an autoimmune aetiology. Acquired isolated or unitrophic ACTH deficiency is a rare but definite cause of adrenal insufficiency. Additional case reports and autopsy studies describing acquired isolated ACTH deficiency associated with autoimmune thyroid disease have appeared in the literature suggesting that the association is more than coincidental. Combined thyroid and adrenal failure should not always be considered to be the result of combined end organ failure. Unitrophic isolated ACTH deficiency may coexist with primary hypothyroidism. PMID- 6280436 TI - Immunoreactive beta-endorphin/lipotrophin in the chronically cannulated ovine foetus: response to bilateral foetal adrenalectomy. PMID- 6280437 TI - The physiology and pathophysiology of the myoneural junction. PMID- 6280438 TI - The peripheral neuropathy in De Sanctis-Cacchione syndrome. Histological, ultrastructural, and morphometric studies. AB - Histological, ultrastructural, and morphometric studies were performed on nerve and muscle biopsies from three patients with de Sanctis-Cacchione syndrome. Sural nerves showed marked loss of the myelinated fibers, in proportion to decrease in nerve conduction velocities and in inverse proportion to the severity of the clinical symptoms, which were related to the survival length. The larger fibers were involved earlier and more markedly than the smaller. The unmyelinated fibers were also decreased in number. Electron-microscopic studies showed the presence of primary degeneration of myelin sheaths or Schwann cells. Muscle biopsies showed grouping of type I and type II fibers in all three patients. Therefore, peripheral nerve involvement in de Sanctis-Cacchione syndrome was suggested to result from chronic degeneration of the neuronal cells and Schwann cells. PMID- 6280439 TI - Postirradiation sarcoma (malignant fibrous histiocytoma) in the inguinal region. AB - A female patient who died of apparent postirradiation sarcoma at the age of 39 was reported. Following a traffic accident, she developed squamous cell carcinoma at the injured right heel 20 years later, and also metastasis to both inguinal and popliteal lymph nodes. During the period from September 1966 to May 1968, she had received Co60 irradiation to the right inguinal and para-aortic areas, 9,600 and 6,600 Roentgen, respectively. Ten years later, she manifested a huge tumor in the inguinal region which extended further to the retroperitoneum; light and electron microscopic examinations proved to be a malignant fibrous histiocytoma. She died of massive pulmonary metastasis two years later. Histological differentiation of the tumor and relationship between irradiation and sarcoma induction were briefly discussed, and the necessity of reevalution of soft part sarcomas was emphasized. PMID- 6280440 TI - Pale eosinophilic inclusions simulating ground-glass appearance of cells of hepatocellular carcinoma. AB - Pale eosinophilic inclusions simulating ground-glass appearance were observed in cells of hepatocellular carcinoma. They were round or elliptical in shape, and their size was about 14 micrometers in diameter. Light eosinophilia and fine granulation with homogeneous appearance were delineated from other cellular materials. They were negative for HBsAg, blood plasma proteins including alpha 1 antitrypsin, and alpha-fetoprotein by immunohistochemical stainings. Ultrastructurally, they were well-demarcated and consisted of homogeneous finely granular matrix. Histological, histochemical and ultrastructural features were different from several intracellular inclusions hitherto reported on hepatocellular carcinoma. These changes may represent a deposition of secretory proteinous materials in cells of hepatocellular carcinoma, but their chemical and antigenetical content could not be identified. PMID- 6280441 TI - [A study of antitumor chemotherapeutic agents--synthesis of hexoestrol dibromoacetate (HL-286) (author's transl)]. PMID- 6280442 TI - [Evidences for possible existence of a "soman receptor" in earthworm dorsal muscle (author's transl)]. PMID- 6280443 TI - [Interaction of seven 3-methyl fentanyl compounds with cerebroside sulfate (author's transl)]. PMID- 6280444 TI - [Effects of 10-hydroxycamptothecin on cAMP and cGMP levels in hepatoma and liver tissue of mice (author's transl)]. PMID- 6280445 TI - Peripheral metabolism of beta-carboline-carboxylic acid esters. AB - Esters of beta-carboline-3-carboxylic acid have recently been identified as potent inhibitors of brain benzodiazepine receptors in vitro. Ethyl beta carboline-3-carboxylate (beta-CCE), however, is a rather weak inhibitor in vivo of benzodiazepine receptors in mice. The ED50-value was 91 mg/kg intraperitoneally 35 min after administration (ED50 is that dose which inhibits by 50% the specific binding of 3H-flunitrazepam intravenously). ED50 for beta-CCE was 2-20 fold lower in mice pretreated with organophosphorus esterase inhibitors, concomitantly with the observation of strong inhibition of liver and kidney hydrolyzing activity, using 3H-propyl beta-carboline-3-carboxylate as substrate. The rat brain contains only approximately 0.1% of the hydrolyzing activity as compared to the liver. It is concluded that some esters of beta-carboline-3 carboxylate exhibit only weak effects on benzodiazepine receptors in living animals due to hydrolysis outside the brain. PMID- 6280446 TI - An improved foot-shock titration procedure in rats for centrally acting analgesics. AB - A foot-shock titration method for the measurement of antinociceptive activity in the rat has been developed. A new grid design is described which makes the use of scrambled shocks unnecessary. Thresholds for responses elicited at different levels of integration within the central nervous system are measured: Detection threshold, flexor reflex, coordinated jumping and vocalization. These thresholds were found to be stable over time in saline-treated rats. The two opioid analgesics morphine and d-propoxyphene increased these thresholds, each in a characteristic way. Morphine was more selective in its action and significantly more effective in increasing the vocalization threshold than the threshold for jumping, whereas d-propoxyphene increased both these thresholds to the same extent. Neither drug affected the detection threshold in low to moderate doses. The difference in pharmacodynamic profile is discussed in terms of heterogeneity of opioid receptor-effector mechanisms at different levels of the neuroaxis. PMID- 6280447 TI - Angiotensin-converting enzyme in hypercalcaemic disorders. AB - Serum angiotensin-converting enzyme (SACE) was elevated (mean +/- S.D. 55.2 +/- 19.8 U/ml) in 11 patients with hypercalcaemia due to sarcoidosis, whereas it was within the normal limits ((20.0 +/- 5.2 U/ml) in 23 patients with other hypercalcaemic conditions. Among these, 16 had primary hyperparathyroidism and a SACE level of 18.6 +/- 4.7 U/ml, significantly lower than in healthy controls (24.4 +/- 6.2 U/ml). In 7 patients with hypercalcaemia due to malignancies or prolonged immobilization SACE was 21.8 +/- 5.9 U/ml. A weakly positive correlation was found between SACE and S-calcium in hypercalcaemic sarcoidosis patients but not in the other patients or in a control group of 144 consecutive sarcoidosis patients. sarcoidosis with hypercalcaemia seems to be associated with a very high prevalence of elevated SACE. Especially when sarcoidosis is suspected to be the cause of hypercalcaemia, SACE measurement may be useful as a rapid diagnostic guide. PMID- 6280448 TI - [Persistence of variola virus in rabbits]. PMID- 6280449 TI - Histoenzmological mapping of ATPase and 5-nucleotidase in the spinal cord and medulla oblongata of hedgehog (Paraechinus micropus). AB - The contribution presented deals with the distribution of adenosine triphosphatase (ATP-A) and 5-nucleotidase (AMP-A) in the spinal cord and medulla oblongata of hedgehog. The highlights of this study are: (1) AMP-A activity is stronger in neuropil than in neurons, in all the areas of spinal cord and medulla oblongata. In the nerve cells the enzyme is localized at the peripheries of the neurons, whereas the cytoplasm and nuclei are completely free from enzymatic activity. Reaction in blood vessels is quite high both in gray and white matter. (2) ATP-A activity is seen mainly at the peripheries of the neurons. The neuropil activity varies from mild to intense. Reaction in blood vessels is quite strong in all the areas. (3) Fibrous bundles and tracts are negative for both the enzymes. (4) In general, the activity of ATP-A and AMP-A is strongest in cranial nerve nuclei, irrespective of their sensory or motor nature. The distribution of these enzymes has been correlated with the functions of various nuclei of spinal cord and medulla oblongata in hedgehog, and compared with other mammals. PMID- 6280450 TI - Granular pituicytomas of the pituitary stalk. PMID- 6280451 TI - Computerized tomography findings in multifocal glioma. AB - Six patients with multifocal glioma are presented. Computerized tomography revealed multiple, discrete, contrast-enhancing lesions in the cerebral hemispheres, suggestive of multiple intracranial metastases. The most accessible lesion was resected at craniotomy in each patient, confirming the diagnosis of primary malignant glioma. Postoperative radiation therapy and chemotherapy were instituted according to current protocols. Since neuroradiological studies may not allow distinction of multifocal glioma from multiple brain metastases, surgical biopsy is suggested in those patients who have no history of cancer. PMID- 6280452 TI - Serum IgM level as an index of malignancy in brain tumours. PMID- 6280453 TI - Malignant fibrous histiocytoma of the temporal bone with intracranial extension. AB - An 18-year-old male presented with a right temporal bone tumour; histology revealed malignant fibrous histiocytoma. Following surgical excision and radiation the tumour recurred. The patient was treated with Vindesine (desacetyl vinblastine amide sulphate) with marked response. Malignant fibrous histiocytomas of the temporal bone are extremely rare; this report suggests that the occasional dramatic response may be obtained by combined modality treatment. PMID- 6280454 TI - Congenital block vertebrae C2-C3 in patients with cervical myelopathy. AB - One hundred and forty-five patients presenting with symptoms of cervical myelopathy were treated surgically. Eight of these patients (5.5%) showed congenital block vertebrae C2-C3. The relationship between this congenital anomaly and the occurrence of spondylotic myelo-radiculopathy is discussed. PMID- 6280455 TI - A comparative study of the treatment of cervical spondylotic myeloradiculopathy. Experience with 50 cases treated by means of extensive laminectomy, foraminotomy, and excision of osteophytes during the past 10 years. AB - This paper reviews management by means of the posterior approach of 50 patients with cervical myeloradiculopathy caused by spondylosis and stenosis of the spinal canal seen in the past 10 years. Careful selection of patients is an absolute necessity since a primary cause of failure occurred in individuals who subsequently proved to have motor neurone disease. Older individuals with long standing neurological deficits, especially long tract signs indicative of fixed lesions, were benefited primarily by a lack of further progression of their disorder and occasional improvement in hand function and gait. Diagnostic evaluation should include electromyography, nerve conduction studies, and sensory evoked cortical potentials. With the introduction of the fourth generation CAT scanning equipment, additional diagnostic information is available regarding the internal configuration of the spinal canal, its contents, and the amount of available space at various levels. Supplementary myelography remains of basic importance. Laminectomy includes two levels above and below the areas of significant canal encroachment. Foraminal decompression with removal of only the innermost third of the foramen permits mobilization of the nerve roots, removal of osteophytes and untethering of the dural sac. A great deal of importance is attached to the preservation of the cervical lordotic curve since, with an adequate decompression and an intact dura, the cord moves dorsally into an expanded canal, rising above the ventrally situated osteophytes. In those patients with reversal of the cervical curve and swan neck deformities, posterior decompression has not been of value. Recent more radical procedures in such cases, such as vertebrectomy, remain to be evaluated. Any procedure which will permit further kyphotic deformity, such as laminectomy, is contraindicated. Eighty-five percent of the patients operated upon by the recommended surgical approach improved. PMID- 6280456 TI - Polyoma T antigens. PMID- 6280457 TI - Regulation of SV40 gene expression. PMID- 6280458 TI - Transformation induced by herpes simplex virus: a potentially novel type of virus cell interaction. PMID- 6280459 TI - The Shope papilloma-carcinoma complex of rabbits: a model system of neoplastic progression and spontaneous regression. PMID- 6280460 TI - The clinical syndrome of adolescent drug abuse. PMID- 6280461 TI - Thymic hormones and the immune system. PMID- 6280462 TI - Biological activities of leukotriene B4. AB - Leukotriene B4 isomer III is released from polymorphonuclear leucocytes, monocytes, eosinophils and macrophages in vitro and has been detected and measured in human synovial fluid in vivo. Its most prominent biological activities are to induce the aggregation of and to stimulate the movement (chemokinesis and chemotaxis) of leucocytes in vitro and it acts as a cytotaxin in vivo. In addition, it increases vascular permeability in vivo when administered together with the vasodilator, PGE2. PMID- 6280463 TI - Aggregation of polymorphonuclear leucocytes (PMNs) by leukotriene B4: effects of cyclooxygenase products and metabolic inhibitors. AB - Leukotriene B4 isomer III (LTB4) stimulates the aggregation of rat PMNs in vitro. The effects of deoxyglucose, iodoacetate, dinitrophenol, cyclohexamide, colchicine, prostaglandins and thromboxane B2 (TXB2) on aggregation were examined. The results demonstrate that, first, aggregation is an active process, the energy being supplied by glycolysis and not mitochondrial respiration. The response can be elicited in the absence of extracellular glucose. Synthesis of protein does not occur during aggregation. An intact microtubular system is required for the expression of a full aggregation response. Prostaglandins E1, E2, F2 alpha and TXB2, products of the cyclooxygenase pathway of arachidonic acid metabolism, partially inhibit LTB-4 induced aggregation by a mechanism which has not yet been elucidated. The prostaglandins and TXB2 themselves do not promote aggregation. PMID- 6280464 TI - Macrophages as targets of inhibitory effects of E-type prostaglandins in immune related inflammation. AB - The anti-inflammatory effects of E-prostaglandins (PGE) are attracting interest because they are mediated through actions on cells which are also targets for the putative immunomodulator functions of PGE. In the majority of experimental inflammatory conditions, in which inhibitory effects of PGE have been demonstrated, a variety of immunocytes are implicated, including diverse lymphocyte populations. The inhibitory effects of PGE, however, are also readily observable on the tissue component of the carrageenin-induced granuloma (an immune-related inflammatory model) in which activated macrophages, but not lymphocytes participate. Granuloma-derived macrophages are particularly interesting cells for the study of responsiveness to PGE, because the results obtained on such cells in vitro are directly related to the anti-inflammatory effects of PGE on the macrophage phase of the granuloma in vivo. Such combined studies have revealed differences between the responsiveness of granuloma macrophages to PGE2 and prostacyclin, while with elicited peritoneal macrophages such a difference could not be observed. The adenylate cyclase in granuloma macrophages appears to be unusually sensitive to activation by PGE2, but insensitive to prostacyclin. PMID- 6280465 TI - Lymphokine-induced macrophage aggregation: involvement of cyclic-GMP and microtubules. AB - Human lymphokine (LK) is known to induce guinea pig macrophage aggregation. This effect can be quantitatively measured with a Born modified platelet aggregometer. This method has been well correlated with the state of delayed hypersensitivity. Previous findings about the mechanism of action of the aggregation indicated that this aggregation is not due to an increase in the cellular level of c-AMP. It is doubtful whether c-AMP is a messenger of the LK action. Using a radio immunoassay, small but significant increases were found in c-GMP levels of LK aggregated macrophages. In addition, exogenous dibutyryl c-GMP and carbamylcholine induced a macrophage aggregation, as did the divalent cation ionophore A-23187. These data, together with the fact that LK-induced macrophage aggregation is inhibited by colchicine and at 0 degree C, suggest that microtubule polymerization is involved in this process. PMID- 6280466 TI - Local control of inflammatory pain. AB - The mechanism of the local hyperalgesic action of prostaglandin E2 has been studied using rat paw oedema. Prostaglandin E2 is a metabotropic transmitter, activating adenylate cyclase, either directly or through the release of a stimulatory protein factor. This activation of adenylate-cyclase is blocked, locally, by opiates. Hyperalgesia results from the alteration of the cAMP/Ca2+ balance after chemical or mechanical initiation of generator potentials at the nociceptor. PMID- 6280467 TI - Detection and quantification of experimental joint inflammation in mice by measurement of 99mTc-pertechnetate uptake. AB - We adapted the method of 99mTc-pertechnetate (99mTc) uptake measurements to the mouse knee for detection and quantification of arthritis because clinical assessment of mouse knee-joint arthritis is not reliable. The main points to ensure reproducibility of measurements were proper fixation and positioning of the knee and careful shielding of the rest of the body from the gamma-radiation detector. 99mTc uptake was calculated as the mean of three countings. The variation coefficient of these countings ranged from 0.007 to 0.082 in non arthritic joints and from 0.025 to 0.081 in arthritic joints. Arthritis was scored as the ratio of the 99mTc uptake in the right knee versus that in the left knee. This ratio averaged 1.06 (S.D. 0.05) in non-arthritic mice 20 minutes after 99mTc administration i.p. On the second day after induction of arthritis in the right knee, this ratio ranged from 1.44 to 1.96; this was significantly higher (P less than 0.005) than in non-arthritic mice, and the increase remained significant during at least 20 days. 99mTc uptake measurements seem to be a useful method to detect and quantify arthritis of the knee joint in mice. PMID- 6280468 TI - Increased incidence and clinical correlation of persistently abnormal technetium pyrophosphate myocardial scintigrams following acute myocardial infarction in patients with diabetes mellitus. AB - "Persistently abnormal" technetium-99m stannous pyrophosphate myocardial scintigrams (PPi+) appear to be associated with a relatively poor prognosis after acute myocardial infarction (AMI). To assess the incidence and implications of PPi+, we performed a retrospective analysis in 29 patients with and 25 patients without diabetes mellitus who had abnormal myocardial scintigrams within 4 days of AMI and who had follow-up scintigrams at least 3 months after hospital discharge. There were no significant differences between patients with and without diabetes as regards age, incidence of transmural or nontransmural AMI, or degree of left ventricular dysfunction after AMI. Persistently abnormal PPi+ occurred more commonly in patients with diabetes than in nondiabetic patients (18 of 29, 62%, compared to 3 of 25, 12%; p less than 0.001). Patients with chronic PPi+ had more frequent cardiac complications following hospital discharge (p less than 0.005) including death, recurrent AMI, unstable angina, and intractable congestive heart failure. Postmortem analysis in two patients with diabetes and chronic PPi+ revealed marked myocytolysis. Thus, patients with diabetes mellitus have an increased incidence of post-AMI "persistently abnormal" technetium (PPi+) scintigrams and relatively poor prognosis following myocardial infarction. PMID- 6280469 TI - Thou, gentle viper: an introduction. PMID- 6280470 TI - Symposium on angiotensin-converting enzyme inhibition: a developing concept. PMID- 6280471 TI - Angiotensin-converting enzyme inhibition: a developing therapeutic concept. Introduction to session I: development. PMID- 6280472 TI - Historical perspectives on the renin-angiotensin-aldosterone system and angiotensin blockade. AB - Advances leading to recognition of the relation of the renin-angiotensin system to aldosterone include: (1) development of analytic techniques for measuring aldosterone, (2) discovery of an aldosterone-stimulating factor in circulating plasma, (3) the finding that a potent aldosterone-stimulating factor is secreted by the kidney, (4) evidence that synthetic angiotensin II increases aldosterone secretion, (5) fractionation of crude kidney extracts and the finding that aldosterone-stimulating factor is renin, (6) the observation that high plasma renin activity occurs in secondary aldosteronism, and (7) recognition that the renin-angiotensin-aldosterone system occurs in congestive heart failure and in renovascular and malignant hypertension. The early use of blocking agents for the renin-angiotensin system is described along with the landmarks of progress. These include the observations that: (1) arterial pressure decreases in experimental renovascular hypertension in response to angiotensin blockade, (2) angiotensin provides important support for arterial pressure in low cardiac output states including congestive heart failure, (3) the kidney participates in this important compensatory mechanism, and (4) cellular receptors for angiotensin are present in the two inner zones of the adrenal cortex. PMID- 6280473 TI - Angiotensin-converting enzyme inhibition and prostaglandins. AB - To determine whether prostaglandins contribute to the depressor response of angiotensin-converting enzyme inhibitors, plasma prostaglandin levels were measured by radioimmunoassay in normo- and hypertensive subjects on both sodium restricted and sodium-loaded diets before and after captopril administration. On the sodium-restricted diet, the hypotensive response to captopril was accompanied by significant increments in the metabolite of prostaglandin E2 (PGE2-M) and bradykinin and by significant decrements in angiotensin II. The high sodium diet suppressed the response of the renin-angiotensin and kinin systems to captopril but the hypotensive response persisted. Furthermore, the decrease in blood pressure correlated significantly with increments in prostaglandin E2-metabolite. Prostaglandin synthesis was then inhibited in the sodium-restricted hypertensive patients by pretreatment with indomethacin. This maneuver completely eliminated the captopril-induced prostaglandin E2-metabolite increment without changing bradykinin or angiotensin II responses but significantly attenuating the hypotensive response. Finally, when patients were studied on a high sodium intake, similar effects were observed except now indomethacin completely abolished the blood pressure response to captopril. These studies therefore support the hypothesis that increased production of vasodilator prostaglandins in a major mediator of the hypotensive response to captopril. Whether the change in prostaglandin release is a direct effect of the drug or secondary to increased kinin levels is uncertain. PMID- 6280474 TI - Angiotensin-converting enzyme inhibition, catecholamines and hemodynamics in essential hypertension. AB - Captopril was given to 15 unselected patients with essential hypertension (WHO II) at a dose range of 300 to 600 mg/day. Hemodynamic indexes (thermodilution) as well as levels of plasma norepinephrine, epinephrine, renin activity and aldosterone were determined simultaneously at the end of 2 weeks of placebo and after 8 weeks of captopril treatment. Systolic and diastolic arterial pressures were reduced significantly by treatment both supine (p less than 0.0025) and standing (p less than 0.0025). The diastolic arterial pressure was normalized (less than 95 mm Hg) in five patients and significantly reduced in four, whereas six patients were considered poor responders (mean arterial pressure decrease 10 mm Hg or less). The decrease in arterial pressure correlated significantly with the reduction in total peripheral resistance (r = 0.71), whereas cardiac index did not change and stroke index increased because of a slight decrease of heart rate. Plasma and urinary norepinephrine and epinephrine did not change during treatment. Moreover, the response of both heart rate and plasma catecholamines to upright posture was not altered by captopril treatment. Plasma renin activity increased and plasma aldosterone concentration decreased during treatment. These results suggest that inhibition of converting enzyme activity by captopril induces a reduction in arterial pressure through a reduction in total peripheral resistance. There was no evidence of an appreciable reduction in sympathetic nervous system activity during therapy. PMID- 6280475 TI - Role of angiotensin II in the physiologic regulation of glomerular filtration. PMID- 6280476 TI - Is angiotensin a prime factor in blood pressure control? PMID- 6280477 TI - Acute and chronic effects of the angiotensin-converting enzyme inhibitor captopril in severe hypertension. PMID- 6280478 TI - In vivo comparison of three orally active inhibitors of angiotensin-converting enzyme. AB - This study was designed to compare the activity of three structurally different drugs (SQ 14225 [captopril], SA 446 and MK 421) as inhibitors of angiotensin I converting enzyme in vivo and to compare their effects on blood pressure in spontaneously hypertensive rats and one clip, two kidney renal hypertensive rats. All the three drugs were potent, orally effective converting enzyme inhibitors. The relative durations of their actions as inhibitors of angiotensin-converting enzyme, from longest to shortest, were as follows: MK 421, SQ 14225 and SA 446. MK 421 appears the most potent on a molar basis. In renal hypertensive rats the drugs appeared equipotent, although the duration of action of MK 421 was prolonged and SA 446 shorter than that of SQ 14225. SA 446 was less effective than the other two compounds in spontaneously hypertensive rats. PMID- 6280479 TI - Long-term effects of captopril on passive sodium permeability and contractility in vascular smooth muscles of spontaneously hypertensive rats. AB - Long-term effects of captopril on passive sodium permeability and cellular sodium concentration in the aorta of spontaneously hypertensive rats were examined in terms of contractile properties of the muscle. In cold lithium solution, cellular sodium in the aorta isolated from spontaneously hypertensive rats leaked more rapidly than did that isolated from Wistar Kyoto rats (WKY), suggesting an increased sodium permeability in the aorta of spontaneously hypertensive rats. Six weeks of treatment with captopril reduced the increased sodium permeability after hydralazine had failed. Inhibition of the sodium pump by incubating the aorta in potassium-free solution produced a slowly developing contraction which was associated with accumulation of cellular sodium. Both the magnitude of contraction and the accumulation of sodium during exposure to potassium-free solution for 1 hour were significantly less in the aorta isolated from spontaneously hypertensive rats treated with captopril compared with those from control spontaneously hypertensive rats. These data suggest that after prolonged administration, captopril alters the abnormal sodium permeability of vascular smooth muscle in spontaneously hypertensive rats and that the restoration of normal permeability reduces vascular tone. PMID- 6280480 TI - Induction of angiotensin I-converting enzyme in rat lung with captopril: the effect of adrenalectomy. AB - In spontaneously hypertensive rats, treatment with captopril, 0.2 g/liter of drinking fluid for 12 to 24 weeks, caused a threefold increase in serum angiotensin I-converting enzyme activity. Angiotensin I-converting enzyme increased 25 to 120 percent in lung plasma membranes. The elution profile of angiotensin I-converting enzyme on DEAE cellulose and after gel filtration on Sepharose 4B was unchanged by captopril. The Km value value also remained unchanged. In Wistar rats subjected to bilateral adrenalectomy, treatment with the same dose of captopril for 3 days resulted in increased serum angiotensin I converting enzyme activity in both sham-operated and adrenalectomized rats, but angiotensin I-converting enzyme concentration increased in lung plasma membranes from sham-operated rats and captopril-treated rats only. We conclude that captopril causes induction of angiotensin-converting enzyme biosynthesis in spontaneously hypertensive and Wistar rats. The change is a quantitative one. Intact adrenal glands may be important for the incorporation of angiotensin I converting enzyme into lung membranes. PMID- 6280481 TI - Angiotensin II is a necessary component for the development of hypertension in the two kidney, one clip rat. AB - The current study was undertaken to define the role of the renin-angiotensin system in the development of hypertension in the two kidney, one clip Goldblatt rat. Captopril was administered orally (100 mg/kg/day) to two groups of rats (n = 8 each) 24 hours before and each day after unilateral renal artery clipping (0.2 mm internal diameter): the drug was given for either 16 weeks (group I) or 24 weeks (group II). Sham-operated (n = 5) and Goldblatt (n = 8) rats not receiving captopril were prepared for comparisons of plasma renin activity and systolic blood pressure. Indomethacin (20 mg/kg/day subcutaneously) was administered for 48 hours concomitantly with captopril to the rats in group I. In group II, systolic blood pressure was monitored for 7 weeks after cessation of captopril. Continual captopril administration to Goldblatt rats completely prevented the rise in systolic blood pressure, a rise that was observed in Goldblatt rats not receiving captopril. Whereas systolic blood pressure of captopril-treated rats approximated 100 mm Hg throughout the study, that of Goldblatt rats not receiving the drug increased to nearly 180 mm Hg within 6 weeks after clipping. Systolic blood pressure of sham-operated rats remained normal. Indomethacin did not change systolic blood pressure in the drug-treated rats in group I. On cessation of captopril therapy in group II, systolic blood pressure increased gradually in a manner that paralleled the development of the disease in the Goldblatt rats that did not receive captopril. Plasma renin activity was determined in Goldblatt and sham-operated rats at either 16 weeks (group I) or 24 weeks (group II) after clipping; the rats from either group with mild hypertension (systolic blood pressure less than 180 mm Hg) had normal plasma renin activity whereas those with severe hypertension (systolic blood pressure greater than 180 mm Hg) had greatly elevated plasma renin activity. In summary, captopril can completely prevent the increase in systolic blood pressure for up to 24 weeks in Goldblatt rats, and this hypotensive effect is not mediated by the prostaglandins. It is concluded that the renin-angiotensin system is a necessary component of the hypertensive process in this experimental model. PMID- 6280482 TI - Dissociation between in vivo and in vitro measurements of converting enzyme activity after chronic oral treatment with captopril in rats. AB - Intravenous administration of captopril (20 micrograms) produced inhibition of angiontensin I pressor responses by 70 percent and of plasma-converting enzyme activity by 72 percent. Oral treatment with captopril (50 mg/kg/day) for 1 week inhibited angiotensin I pressor responses more (84 percent) than plasma converting enzyme activity (23 percent). Four month oral treatment of normotensive and spontaneously hypertensive rats with captopril (50 mg/kg/day) led to 68 and 71 percent inhibition of angiotensin I pressor responses, but produced increases in plasma-converting enzyme activity of 123 and 94 percent, respectively. In spontaneously hypertensive rats, elevated converting enzyme activity in the medulla oblongata was measured after this treatment. It is concluded that plasma-converting enzyme activity measurements can be dissociated from the in vivo inhibition of converting enzyme. Chronic oral captopril treatment results in an induction of converting enzyme biosynthesis not only in peripheral tissue but also in the brain. PMID- 6280483 TI - Comparative effects of captopril and MK 421 on sympathetic function in spontaneously hypertensive rats. AB - The effects of captopril and MK 421, both orally active angiotensin-converting enzyme inhibitors, on sympathetic function in the pithed spontaneously hypertensive rats were examined. Captopril (100 mg/kg orally) significantly inhibited pressor responses to sympathetic nerve stimulation and norepinephrine whereas MK 421 (10 and 100 mg/kg orally) was without effect. Both drugs abolished the angiotensin I response without affecting that to angiotensin II. Because these doses of MK 421 are antihypertensive in the spontaneously hypertensive rat, it is concluded that inhibition of sympathetic function plays no role in its effect. The role of the sympatholytic action of captopril in its antihypertensive action remains unclear. PMID- 6280484 TI - Comparison in normal volunteers of three converting enzyme inhibitors: RHC 3659, MK 421 and captopril. PMID- 6280485 TI - Acute hypotensive and renin-stimulating actions of captopril before and during treatment with a beta-blocking drug. AB - There is no general agreement on the relation between the hypotensive effect of captopril and the pretreatment plasma renin levels of hypertensive patients. To determine whether the hypotensive effect of captopril was directly related to plasma renin, the angiotensin-converting enzyme inhibitor was administered acutely to 10 essential hypertensive patients with normal or suppressed plasma renin activity before and after inhibition of renin secretion with propranolol. Captopril was equally effective in reducing blood pressure both when administered alone (25 mg: -29/-17; 50 mg: -37/-23 mm Hg) and after chronic treatment with propranolol (25 mg: -33/-20; 50 mg: -30/-20 mm Hg). The increase in renin induced by captopril was not decreased by propranolol therapy. The persistence of the hypotensive effect of captopril after renin suppression by propranolol suggests that this drug has some blood pressure decreasing properties independent of plasma renin. PMID- 6280486 TI - Humoral and hemodynamic effects of increasing doses of captopril in patients with essential hypertension. AB - To compare the hemodynamic and humoral effects of increasing doses of captopril, blood pressure, heart rate, plasma renin activity, plasma aldosterone and angiotensin-converting enzyme activity were measured in 10 patients with mild uncomplicated essential hypertension before and after captopril given in an increasing dose of 25, 50 and 100 mg twice a day, with each dose given for 3 days. The maximal decrease in blood pressure, which was predicted both by basal plasma renin activity values and by the hypotensive response to the first dose, was found after the lowest (25 mg) dose; this effect was detectable for 12 hours independently of the dose administered. Similarly, plasma renin activity was already maximally increased and plasma aldosterone maximally decreased at the lowest dose, while angiotensin-converting enzyme activity showed a dose-dependent inhibition. These data suggest that (1) neither the magnitude nor the duration of the hypotensive effect nor the resin-stimulating and aldosterone-inhibiting actions of captopril are enhanced by drug doses above 25 mg (at least up to 100 mg), and (2) there was no apparent correlation between the degree of angiotensin converting enzyme inhibition with increasing doses of captopril and the hypotensive effects of the drug. PMID- 6280487 TI - Sequence of development of iron deficiency in the rat. AB - This study was designed to determine the interrelationships among storage iron, transport iron, and iron compounds that serve known physiological functions (Hb, myoglobin, and cytochrome c) during the gradual progression of dietary iron deficiency. These three categories of iron compounds are generally considered to become depleted in three corresponding, sequential stages. However, there is scattered evidence of substantial overlap between these stages in man. The presence of such overlap may prove pertinent to the interpretation of laboratory tests used in the diagnosis of iron deficiency. The rat was used as an experimental model to allow more complete evaluation of the interrelationships between the stages of iron deficiency than would be possible in man. Rats were given diets containing 2, 6 and 50 mg iron/kg diet during early adult development, between 36 and 90 days of age. The iron-deficient diets (2 and 6 mg iron/kg diet) resulted in decreases in hematocrit and in muscle and intestinal cytochrome c well before storage iron in the liver and spleen was exhausted. The results in the rat model may help to explain why there is not a consistent pattern of laboratory abnormalities in individuals with chronic, mild dietary iron deficiency. PMID- 6280488 TI - Copper bioavailability and requirements. AB - Knowledge of factors affecting the bioavailability of dietary copper is limited. Intestinal absorption of copper appears to be facilitated by L-amino acids. Picolinic acid has a favorable binding affinity for copper and may facilitate its absorption. Measurements of the dietary requirements for copper in adult men have shown the requirement to range from about 1.5 to 2.0 mg daily, levels similar to the 2.0 mg estimate suggested in the past. Comparison of the copper requirements with the levels of copper present in some contemporary diets suggests that marginal copper nutriture may not be rare. Persons who consume diets high in zinc and low in protein are at risk of copper deficiency. High intakes of sources of dietary fiber apparently increase the dietary requirement for copper. Studies in one man have shown that signs of mild copper deficiency can be produced experimentally when a conventional diet containing about 0.8 mg of copper is fed. At this time, the 2 to 3 mg daily intake of dietary copper suggested by the National Research Council (63) seems appropriate. PMID- 6280489 TI - The launching of a cytomegalovirus vaccine. PMID- 6280490 TI - Vaccination of pediatric nurses with live attenuated cytomegalovirus. AB - Congenital cytomegalovirus (CMV) infections contribute considerably to morbidity and mortality among infants in the United States. In a preliminary study aimed at protecting fetuses from congenital disease we evaluated Towne 125-strain CMV vaccine in ten female pediatric nurses of childbearing age. The women were seen 2, 4, 8, 12, 26, 39, and 52 weeks after vaccination; specimens were obtained for routine laboratory testing, virus isolation, cell-mediated immunity testing, and serologic examination (complement fixation, anticomplement immunofluorescence [ACIF], and neutralization). Local but not systemic reactions were observed in all subjects. Serologic responses appeared at two to four weeks and peaked at four to eight weeks. Mean titers were highest in the ACIF test. A cell-mediated immune response, as assayed by lymphocyte proliferation, was observed in all women, and in most there was a biphasic pattern. Towne 125-strain CMV vaccine seems to be safe and immunogenic in women of childbearing age. PMID- 6280491 TI - Fetal and maternal virilization associated with pregnancy. A case report and review of the literature. AB - A masculinized female infant was born to a mother who had virilizing signs dating from the fourth month of pregnancy. Serum 17 alpha-hydroxyprogesterone, dehydroepiandrosterone, and testosterone levels were all normal in the infant. Maternal testosterone level was markedly elevated one week post partum. Dexamethasone phosphate suppression was normal. Human chorionic gonadotropin stimulation five weeks post partum revealed further elevation of high baseline free and total testosterone levels. Free and total testosterone levels 30 weeks post partum were normal, and all maternal virilizing signs had regressed with the exception of her deepened voice. The child has had no progression of masculinization. The mother is believed to have had a luteoma of pregnancy. PMID- 6280492 TI - Perspectives on marijuana use and abuse and recommendations for preventing abuse. AB - Survey research on levels of use and attitudes toward marijuana by youth indicate a consistently favorable attitude toward occasional, moderate use. Use of marijuana is contrasted with the use of alcohol and tobacco by youth. Research findings re the dangers of marijuana use are summarized. The inability of legislators and law enforcement personnel to control access to and use of marijuana, coupled with the high cost of our present legal response to the marijuana challenge, leads to recommending legalization of marijuana, as well as, developing a generic primary prevention program for marijuana abuse which features distinguishing "responsible use" from "abuse." Criteria for judging marijuana abuse are provided. PMID- 6280493 TI - Antibody-dependent cell-mediated cytotoxicity to herpes simplex virus-transformed cells in the course of cervical carcinoma. AB - The antibody-dependent cell-mediated cytotoxicity assay was used to determine whether humoral antibodies from women with cervical carcinoma in the presence of normal mononuclear cells could induce cytotoxicity to hamster embryo fibroblasts, originally transformed by herpes simplex virus type 2 (HSV-2) (333-8-9), to a cervical carcinoma cell line (ME-180) and to a lung carcinoma cell line (A-549). Control groups consisted of 81 age-matched healthy women and 77 patients suffering from malignancies other than cervical carcinoma. Significant cytotoxicity of HSV-2-transformed cells was observed in sera from 20 of 28 (71%) surviving patients with cervical cancer who were followed with serum samples collected 6-24 months after initial treatment. Comparison of serum samples obtained before initial treatment from 20 patients surviving the observation period of 60 months or longer and from 20 patients who died during the observation period showed that the survivors had significantly higher cytolytic activity prior to treatment. Long-term survivors also showed a drop in cytolytic activity in sera obtained 30 months or later after treatment. Patients with long survival but whose cancer finally caused death, had stronger cytolytic activity than those with short survival (24 months or less). Patients with cervical cancer had significantly higher cytolytic activity to HSV-2-transformed cells and to the cervical carcinoma cells (ME-180) than age-matched control women and patients with malignancies other than cervical carcinoma. PMID- 6280494 TI - A population study of herpesvirus infections and HLA antigens. AB - In a population-based study of herpesvirus infection and human leukocyte antigen (HLA) phenotype, conducted during the period 1977-1979 in the Framingham Study cohort, HLA-Bw16 was found less often and HLA-Cw2 more often in individuals with histories of herpes labialis than in other individuals. Similarly, both geometric mean antibody titer to herpes simplex type 1 and proportion of individuals with detectable antibody were higher in those without Bw16 and in those with Cw2. No strong association was found between HLA and history of herpes zoster or titer to herpes simplex type 2, varicella-zoster virus or cytomegalovirus. PMID- 6280495 TI - Morphologic observations of contact-induced lysis of EBV-infected B lymphocytes by autologous hand mirror T cells. AB - To study the possible immunologic role of hand mirror lymphocytes (HML) in the control of Epstein-Barr virus (EBV)-infected B lymphocytes in patients with infectious mononucleosis (IM), we studied the in vitro interactions of these cells by time-lapse video light microscopy. Peripheral blood T lymphocytes isolated from three patients in the early recovery phase of IM were mixed with their autologous EBV-infected B cells. Motile lymphocytes with their characteristic hand mirror shape were observed to attach by their uropods to the B cells. The HML remained attached for variable periods ranging from 45-75 min. Following detachment, B cells that were in contact with HML underwent lysis. Mixtures of T cells from healthy donors and EBV-infected cell lines exhibited only rare uropod formation with no attachment or lysis of B cells. The present experiment indicates that contact-induced lysis of EBV-infected B lymphocytes is operative in IM and that this process is mediated by the HML. PMID- 6280496 TI - Assignment of first random restriction fragment length polymorphism (RFLP) locus ((D14S1) to a region of human chromosome 14. AB - A locus responsible for a restriction fragment length polymorphism (RFLP) has been identified by hybridization of Eco RI fragments to the random human DNA sequence in recombinant plasmid pAW101. We have examined DNA extracted from 20 human X Chinese hamster somatic cell hybrids for the presence of sequences homologous to the human insert in pAW101. The hybrids were derived from six different human donors, five of whom were heterozygous, producing two bands on Southern transfers. The presence of homologous sequences in the hybrids correlated exclusively with the presence of human chromosome 14. Three hybrids contained chromosome 14 in a frequency of greater than one per cell and were positive for two alleles. Two hybrids contained only the distal half of the long arm of 14 as part of a translocation and were still positive. These results assign the first highly polymorphic random RFPL locus (D14S1) to region q21 leads to qter of chromosome 14. PMID- 6280497 TI - Possible interaction of cefotaxime with a glucose-testing method. PMID- 6280498 TI - Outbreak of Kaposi's sarcoma with cytomegalovirus infection in young homosexual men. AB - Kaposi's sarcoma, a multicentric malignant neoplasm, occurs in certain geographic areas in the world. It is most common in Equatorial Africa and Eastern Europe. The annual incidence of Kaposi's sarcoma in the United States is between 0.021 and 0.061 per 100,000 persons. The appearance of an outbreak of Kaposi's sarcoma in young homosexual men in New York and California is a new and unique phenomenon. Certain differences are already recognized between the disease in these young men and the ordinary Kaposi's sarcoma. Herein we report our observations of the first 10 cases of Kaposi's sarcoma in young homosexual men. In these patients, the disease follows an aggressive clinical course characterized by widespread skin lesions with early involvement of the lymph nodes. In some of these patients, the result was death in a short period of time after initial diagnosis. In addition, cytomegalovirus infections were seen in these patients, which suggests at least a possible association between this viral and the disease. PMID- 6280499 TI - Ketoconazole treatment of nonprimary coccidioidomycosis. Evaluation of 60 patients during three years of study. AB - Sixty patients with coccidioidomycosis were treated with ketoconazole rather than with another antifungal agent, and their responses were evaluated in relation to the predominant site of involvement. For the three main groups, improvement occurred in 12 of 19 patients with chronic pulmonary infections, in 20 of 23 with soft tissue lesions and in six of 11 with skeletal involvement. Infections in soft tissues improved most rapidly (average of 34 days) and often with 200 mg per day, whereas pulmonary and skeletal infections improved more slowly (63 and 165 days, respectively), usually requiring 400 mg per day. Of 12 patients with response in whom therapy has been discontinued, seven have had relapses. Recurrence was apparent usually within the first month and after six months or less of treatment. Patients in remission had received ketoconazole for six to 17 months. Untoward drug effects included abdominal complaints (23 percent) and gynecomastia (8 percent). Therapy was discontinued in only three patients for side effects. Our findings support the use of ketoconazole in the treatment of certain forms of chronic coccidioidal infections. PMID- 6280500 TI - Diagnosis of severe fetal cytomegalovirus infection from amniotic fluid in the third trimester of pregnancy. PMID- 6280501 TI - Pregnancy outcome subsequent to consecutive hydatidiform moles. PMID- 6280502 TI - Vaginitis in sexually active women: relationship to nine sexually transmitted organisms. AB - Women seen for symptoms suggestive of vulvovaginitis were studied for the detection of Mycoplasma hominis. Ureaplasma urealyticum, yeast, Neisseria gonorrhoeae, Chlamydia trachomatis, Gardnerella vaginalis, herpes simplex virus, group B beta-hemolytic streptococci, aerobes, anaerobes, and Trichomonas vaginalis. Asymptomatic women who reported to be sexually active and agreed to undergo comprehensive genital cultures were used as controls. There was a significant association of vulvovaginitis with the recovery of sexually transmitted organisms. However, all organisms were also recovered from asymptomatic patients. A total of 468 sexually transmitted organisms were recovered from 253 symptomatic patients (1.85 organisms per patient), while 125 were recovered from 130 asymptomatic patients (0.96 organisms per patient). The difference in prevalence between symptomatic and asymptomatic women for Chlamydia trachomatis, group B streptococcus, and the mycoplasmas was not statistically significant. Factors that predispose patients to the manifestations of symptoms are not clearly understood. PMID- 6280504 TI - Fibrous histiocytoma of the lacrimal sac. AB - Fibrous histiocytoma is a rare tumor whose ocular manifestations usually involve the orbit or, less commonly, the conjunctiva and ciliary body. We have treated two patients with fibrous histiocytoma of the lacrimal sac. One, a 62-year-old woman who had had hordeolum and trachoma, had a visual acuity of counting fingers at 1 m in the affected eye. The fellow eye had been enucleated ten years previously. The entire lacrimal sac was surgically removed and a brown cystic tumor measuring 28 x 12 x 10 mm was found. The second patient, a 32-year-old man, had undergone an unsuccessful dacryocystectomy for epiphora. When he underwent a dacryocystorhinostomy some months later, a mass measuring 20 x 15 x 12 mm was found in the wall of the lacrimal sac. Microscopic examination of the two excised lacrimal sacs showed that the walls were thickened by cells resembling fibroblasts and by cells resembling histiocytes. The fibroblasts were characterized by collagen production, were fusiform or oval in shape, and were arranged in bundles. The histiocytes were larger and had abundant (and sometimes vacuolated) cytoplasm. All the cells appeared to be mature. PMID- 6280503 TI - Ocular disorders associated with a new severe acquired cellular immunodeficiency syndrome. AB - Among the prominent features of a newly described acquired cellular immunodeficiency syndrome that affects previously healthy male homosexuals are multiple opportunistic infections and Kaposi's sarcoma. Immunosuppression induced by cytomegalovirus infection may play a major role in the pathogenesis of this disorder. We have performed ophthalmic examinations on seven such patients and found ocular abnormalities in all cases. Six patients were examined during the course of their illness and one patient at autopsy only. Each patient had several cotton-wool spots in the affected eye. Other ocular findings included cytomegalovirus retinitis, severe retinal periphlebitis, and conjunctival Kaposi's sarcoma (one case each). PMID- 6280505 TI - Assessing patient satisfaction with state hospital and aftercare services. AB - To assess patients' satisfaction with state hospital and aftercare services, the authors administered an open-ended interview to 22 former state hospital patients. The patients expressed marked dissatisfaction with their treatment in the hospital, but 78% said they were helped by the aftercare services. Important components of their satisfaction were feeling helped, feeling understood, and feeling that the aftercare worker was available when needed. Frequency and length of meetings and types of assistance were not related to satisfaction. The patients reported that the psychotherapeutic and task-oriented assistance were equally helpful. PMID- 6280506 TI - Rotavirus infection in Nigerian infants and young children with gastroenteritis. AB - In a study conducted over a 5-month period (September 1979 to January 1980), rotaviruses were detected by counterimmunoelectrophoresis in the feces of 16 (13.8%) of 116 infants and young children between 1 and 48 months old with acute, sporadic gastroenteritis, at Ife, Nigeria. The highest frequency of rotavirus infection was found in th 7- to 12-month age-group (18.9%). Males constituted 61% of the sample and excreted rotavirus at a significantly higher rate than did females (P less than 0.01). Rotavirus infection was higher in the drier months of November-January (19.3%) than during the rainy months of September and October (8.5%). PMID- 6280507 TI - Significance of clostridial bacteremia. PMID- 6280508 TI - Minimal brain dysfunction in adolescents and young adults: diagnostic and therapeutic perspectives. PMID- 6280509 TI - Evoked brain potential deficits in alcoholism and aging. PMID- 6280510 TI - The use of agar for gel electrophoresis of DNA. PMID- 6280511 TI - Radioimmunolocalization of murine mammary tumor virus proteins in gels. PMID- 6280512 TI - A simple and rapid method for the synthesis of nucleoside 5'-monophosphates enriched with 17O or 18O on the phosphate group. PMID- 6280513 TI - Beta-Galactosidase immunoassay for the measurement of cyclic AMP. PMID- 6280514 TI - A novel reversible thiol-specific spin label: papain active site labeling and inhibition. PMID- 6280515 TI - [Investigations on the ultrastructural cytology of branchiomeric paragangliomas (author's transl)]. AB - The paragangliomas of the jugular and carotid bodies are growth processes which show varying histological and cytological patterns. The real nature of these lesions is still a matter of discussion. In this study 6 jugular and 3 carotid body tumours were investigated using the electron microscope. The main components were irregular cells with short and long cytoplasmic processes closely packed and mostly arranged in complexes. The ultrastructural cytology of the jugular and carotid bodies showed no striking differences. The great majority of the cells exhibits a clear similarity to the cells of normal paraganglia, especially to chief cells of carotid bodies (type I cells). But there were some modifications in the cellular structure including the abnormal structure of mitochondria and a greater variation in the morphology of the dense cored granules. The cytoplasm was dominated by differing populations of mitochondria comprising such of the crista type and such showing tubulus-like structures with a dense matrix and very often intramitochondrial bodies. The characteristic feature was the occurrence of randomly distributed dense cored granules measuring 700-800 A in diameter. These granules display a homogeneous electron dense or moderately dense somewhat granular core which is separated from limiting membrane by a clear electron lucent halo. A further population of granules was of a greater dimension (3,000 4,000 A) revealing an oval or cigar-like shape. In contrast to the former type the homogeneous dense cores of these granules showed a smooth contour and the lucent halo was very narrow. In general, singular exocytotic phenomena could be supposed. More frequently, however, signs of granulolysis were encountered. Free ribosomes, arranged often as polysomes, sparse, disorderly distributed microfilaments of 60-80 A diameter, few 240 A-microtubules and some lysosomal structures are also observed. Usually the cell clusters abutted directly upon the vascular connective tissue. In other cases cytoplasmic processes are intercalated resembling sustentacular cells. Fenestrated capillaries known to be typical for the paraganglionic tissues were only seldom detected. In the interstitial tissue typical collagen fibres are found showing a periodicity of 600-700 A, but occasionally fibres with a periodicity of about 1,200 A could be seen resembling fibrous-long-spacing collagen. Synaptic junctions and nerves or axons were always lacking. PMID- 6280516 TI - Binding of halothane-free radicals to fatty acids following UV irradiation. PMID- 6280517 TI - Adrenocortical suppression in cats given megestrol acetate. AB - Megestrol acetate was given orally to 8 cats at a dose of 2.5 mg every other day for 2 weeks and to 8 cats at a dose of 5.0 mg every day for 2 weeks. Four cats were designated nontreated controls. Pre-ACTH-stimulated plasma concentrations of cortisol (hydrocortisone) and ACTH-stimulated cortisol and tolerance to large dose glucose infusion (IV) were determined on each of the 20 cats given megestrol acetate. Cats were restrained with acepromazine maleate and ketamine hydrochloride during blood sample collection and large-dose glucose infusion. Adrenocortical function and tolerance to large-dose glucose infusion were reevaluated for 4 weeks--after 1st and 2nd weeks of megestrol acetate treatment of the treated groups, and after 1st and 2nd weeks when treatment was stopped (ie, experiment weeks 3 and 4). Each week a cat from the control group and 2 cats from the 2 treated groups were selected to determine the changes occurring during the experiment for that week; after collection of plasma samples, each week's 5 selected cats were euthanatized and necropsied. Significant impairment of adrenocortical function and alteration of adrenocortical morphology occurred with both treated groups. The most severe adrenocortical alterations occurred in the cats 1 week after megestrol acetate was no longer given (ie, experiment week 3). Megestrol acetate-induced adrenocortical suppression contributed to the death of 1 cat. It was concluded that if stress occurs to cats on treatment or soon after treatment with megestrol acetate, glucocorticoids should be supplemented. The effects of megestrol acetate on glucose tolerance were overshadowed by the unforeseen intolerance caused by chemical restraint with acepromazine maleate and ketamine hydrochloride. PMID- 6280518 TI - Pathogenesis of feline infetious peritonitis: pathologic changes and immunofluorescence. AB - Feline infectious peritonitis (FIP) was experimentally induced in FIP virus (FIPV) antibody-positive and antibody-negative kittens after challenge exposure to live-virus aerosol. Seropositive kittens developed antiviral immunofluorescence and lesions more rapidly after challenge exposure than did seronegative kittens. In seropositive kittens, FIPV antigen was present in macrophages and large mononuclear cells in tracheobronchial lymph nodes, lungs, and trachea on postchallenge-exposure day (PCD) 2; in liver and spleen on PCD 3; in kidneys and omentum on PCD 4; and subsequently in nasal turbinates, thoracic and abdominal lymph nodes, thymus, bone marrow, parotid salivary gland, eyes, and brain. Initial antiviral immunofluorescence on PCD 2 coincided with the onset of viremia and vascular lesions. Systemic lesions characterized by perivascular necrotizing pyogranulomatous inflammation, phlebitis and thrombosis, fibrinous serositis, and generalized lymphoid necrosis developed on PCD 3 and 4. Coronavirus-like particles were observed by electron microscopy in cytoplasmic vacuoles or smooth endoplasmic reticulum of degenerating macrophages in inflammatory lesions. In seronegative kittens, antiviral immunofluorescence in tracheobronchial lymph nodes was first detected on PCD 5, and viremia occurred on PCD 6. Systemic necrotizing lesions, comparable with those observed in seropositive kittens on PCD 3 or 4, did not occur in seronegative kittens until PCD 13 or 16. In both groups of kittens, initial viral infection in regional lymphoreticular tissue was followed by viremia and infection of macrophages in reticuloendothelial organs (liver, spleen, lymph nodes) and perivascular locations. The accelerated onset of infection and lesions indicative of an Arthus type reaction in challenge-exposed seropositive vs seronegative kittens further supports the immune-mediated pathogenesis of FIP. PMID- 6280519 TI - Concentrations of corticosteroids, leukocytes, and immunoglobulins in blood and milk after administration of ACTH to lactating dairy cattle: effects on phagocytosis of Staphylococcus aureus by polymorphonuclear leukocytes. AB - A study was conducted to determine relationships among plasma and milk corticosteroids, milk immunoglobulins, and phagocytosis of Staphylococcus aureus by polymorphonuclear leukocytes (PMN) isolated from milk of cows given injections of 0 (saline solution only), 100, and 200 IU of ACTH. Also determined were the effects of ACTH on mobilization of PMN into milk from the mammary gland irritated by infusion of 100 ml of saline solution containing 0.1% oyster glycogen. Cows (n = 10) were injected 6 times within a 48-hour period with 0, 100, or 200 IU of ACTH. Immediately before cows were given the 5th injection, 2 mammary quarters were infused with the saline-glycogen solution. Five hours after the 6th injection, milk from infused quarters was collected, and PMN were isolated, washed, and resuspended in autologous skimmed milk collected 5 hours after the 4th injection and before the udder was infused. Isolated PMN were incubated with S aureus and percentage of phagocytosis was determined. Concentrations of total corticosteroids in plasma and milk increased after cows were given injections of 100 and 200 IU of ACTH. The concentrations of IgA, IgG1, IgG2, IgM, and bovine serum albumin in milk after 4 injections of 0, 100, and 200 IU of ACTH were similar to preinjection (base line) concentrations. Injections of 100 and 200 IU of ACTH significantly increased the concentrations of total circulating leukocytes. Concentrations of leukocytes in milk tended to increased with increasing doses of ACTH, but the differences were not significant. Injection of 100 and 200 IU of ACTH reduced phagocytosis of S aureus by PMN. After 60 minutes of incubation, phagocytosis averaged 57% and 54%, respectively, for the ACTH treatments and 70% for controls (saline only). Results indicate that injections of ACTH that result in physiologic and pharmacologic plasma concentrations of corticosteroids inhibit phagocytosis. Impairment of phagocytosis appeared to be a direct effect of corticosteroid concentration o PMN and was not due to reduced concentrations of immunolobulins. These data indicate that reduced phagocytosis by PMN could be important in increased susceptibility to disease during stress in lactating dairy cows. PMID- 6280521 TI - Sequential mitogen stimulation of peripheral blood lymphocytes from chickens inoculated with infectious bursal disease virus. AB - Chickens were inoculated wih the pathogenic Edgar strain of infectious bursal disease virus at 1 week, 2 weeks, or 1 day of age. In the 3 experiments, phytohemagglutinin stimulation of peripheral blood lymphocytes was significantly decreased on day 3 or 4 after inoculation. Subsequently, on days 7 through 21, stimulations were similar between lymphocytes from inoculated birds and those from control birds. Pokeweed mitogen stimulation was affected minimally in virus inoculated chickens. In each experiment, on day 7, the spontaneous [3H]thymidine uptake was greater in nonstimulated lymphocyte cultures from inoculated chickens than in such cultures from control chickens. In an additional experiment, chickens 1 week of age were exposed to a pathogenic vaccinal virus given in their water. The vaccinal virus exposure resulted in significant decrease of phytohemagglutinin stimulation of lymphocytes on days 3 and 7 of the experiment. A significant decrease in pokeweed mitogen stimulation was observed on day 10 after inoculation. PMID- 6280520 TI - Serologic and molecular comparisons of several equine herpesvirus type 1 strains. AB - The molecular and serologic relatedness of 2 recent respiratory tract isolates of equine herpesvirus type 1, designated T1 and T2, were compared with the Army 183, Kentucky-A hamster-adapted (KyA-ha), and L-M cell-adapted (KyA-LM) strains. Electrophoresis in polyacrylamide gels revealed differences in virion structural proteins among 4 purified strains. Seven envelope glycoproteins (molecular weight of 93,000, 65,000, 62,000, 60,000, 36,000, 20,000, and 18,000) corresponding to virion proteins 13, 16, 17, 18, 23, 25, and 26a, respectively, found in both the Army 183 and KyA-ha strains had slightly different molecular weight counterparts in both the T1 and T2 isolates, which had identical structural protein profiles. virion protein 19 (58,000 daltons), a nonglycosylated protein, was present in reduced amounts in the respiratory tract isolates, whereas virion protein 8a (200,000 daltons) was absent. Virion protein 8a, an envelope glycoprotein, was only present in the KyA-ha strain. The T1 and T2 isolates were not neutralized by equine herpesvirus type 2 antiserum and revealed little cross-neutralizatio with the Army 183 and KyA-ha strains in plaque-reduction neutralization tests. Restriction endonuclease cleavage maps of viral DNA revealed a similar, but not identical, number and size of DNA fragments between T1 and T2 isolates. Likewise, DNa profiles of Army 183, KyA-ha, and KyA-LM were also similar to each other, but vastly different from the respiratory tract isolates. PMID- 6280522 TI - Routine isolation and cultivation of bovine rotaviruses in cell culture. AB - Using the bovine embryonic kidney cell line, Aubek, bovine rotaviruses were routinely isolated from fecal samples of calves with diarrhea. Of 125 fecal samples positive for rotavirus by immune electron microscopy and the enzyme linked immunosorbent assay, 61 isolates were recovered and cultivated continuously. PMID- 6280523 TI - Bacteriologic examination of equine fecal flora as a diagnostic tool for equine intestinal clostridiosis. AB - The fecal flora of 56 clinically healthy and 23 sick horses were examined bacteriologically for counts of Clostridium perfringens, molds, coliforms, alpha- and beta-hemolytic streptococci, and microbes belonging to genus Bacillus, as well as for the presence of Salmonella spp. Of the healthy horses, 85.7% had a C perfringens count less than 10(1) colony-forming units/g of feces. Of the healthy horses, lowest counts were found in race-horses. Of the sick horses, equine intestinal clostridiosis was diagnosed in 2 horses with large C perfringens counts (10(4) to 10(7) colony-forming units/g) and with acute diarrhea. The 7 isolates of C perfringens were identified as serotype A. Salmonella spp were not detected from any of the horses. The study indicated that diagnosing equine intestinal clostridiosis based on the determination of the fecal C perfringens count was suitable. PMID- 6280524 TI - Experimental maedi virus infection in sheep: early cellular and humoral immune response following parenteral inoculation. AB - The early immune response (19 weeks) in sheep inoculated parenterally with maedi virus was studied, using the lymphocyte transformation test, immunodiffusion test, complement-fixation test, and neutralization test. Cellular immune responses were detected between 3 and 7 weeks after the sheep were inoculated. Antibodies were detected by immunodiffusion test in 3 of 5 animals in weeks 7 to 11 and by complement-fixation test in weeks 15 to 19. Neutralizing antibodies were not detected. There was no sign of virus-induced immunosuppression as determined by lymphocyte responsiveness to mitogens or by in vivo and in vitro immune responses following BCG vaccination. PMID- 6280525 TI - Experimental maedi virus infection in sheep: cellular and humoral immune response during three years following intranasal inoculation. AB - A long-term experiment in sheep inoculated intranasally with 2 strains of Norwegian maedi virus was carried out in 2 groups of Norwegian Dala sheep (7 sheep/group). Virus-specific cellular immune response was assayed in the lymphocyte transformation test sequentially during 3 years after sheep were inoculated in group 1 and 4 times in the 3rd year in group 2. Humoral immune response was assayed by immunodiffusion, complement-fixation, and neutralization tests on sequential serum samples collected from the 2 groups. Attempts to isolate virus were made. All group 1 sheep showed transient and irregularly recurring cellular immune responses. In group 2, 6 of the 7 sheep gave similar responses. The frequency of virus isolations was low compared with that reported by various research workers using other breeds for studying experimental maedi visna infection. Precipitating antibodies were detected earlier, and in more animals, than were complement-fixing antibodies. Both were, however, detected later and less frequently than were reported by other research workers. There was a marked difference in the capability of the 2 maedi virus strains to induce neutralizing antibodies. The sequential sera usually showed distinct differences in neutralizing capacity of the virus strains, indicating that they are antigenically different. PMID- 6280526 TI - Effects of ACTH administration on bovine polymorphonuclear leukocyte function and lymphocyte blastogenesis. AB - Yearling steers were treated with ACTH to determine the effect of increased plasma cortisol concentration on bovine lymphocyte and polymorphonuclear leukocyte (PMN) function. The administration of ACTH caused a significant (P less than 0.01) increase in serum cortisol concentration and depression of lymphocyte blastogenesis in response to phytohemagglutinin and concanavalin A. The response to pokeweed mitogen was also depressed, but not significantly. Random migration by PMN was significantly enhanced by ACTH treatment, but there was no effect on ingestion of Staphylococcus aureus, nitroblue tetrazolium reduction, or antibody dependent cell-mediated cytotoxicity by PMN. The iodination reaction, which evaluates the activity of the myeloperoxidase-hydrogen peroxide-halide antibacterial system of the PMN, was significantly impaired after ACTH treatment. These data indicate that specific parameters of lymphocyte and neutrophil function were impaired directly or indirectly by elevated in vivo concentrations of plasma cortisol. PMID- 6280527 TI - Bluetongue virus in pregnant elk and their calves. AB - Two pregnant North American elk (Cervus canadensis), in the 3rd and 4th months of gestation, were inoculated with bluetongue (BT) virus (BTV) serotype 11. The virus was not isolated from the blood of the cows beyond postinoculation day (PID) 8, but was isolated from bone marrow and spleen samples obtained at necropsy on PID 190. Although neither cow had overt clinical signs of BT infection, fluctuations in specific neutralizing BTV antibody titers indicated viral replication. However, in 2 attempts, BTV was not recovered biologically via bites of colonized Culicoides variipennis (biting gnats) with subsequent transmission of the BTV to sheep. Bluetongue virus was isolated from the elk calves at birth and before they nursed. These calves remained latently infected, and BTV was transmitted from each calf to sheep by bites of the biting gnats. Most of the BTV biological recovery attempts resulted in suspicious BT clinical responses in sheep, but without viral isolation. However, after challenge exposure with the homologous virus, 5 of 7 recipient sheep bitten by the gnats reacted with an intensified BT clinical response that indicated viral sensitization. One calf was born weak, never attained a healthy appearance, was latently infected with BTV, and had fluctuating BTV neutralizing antibody titers. The other calf was in apparently good health, was latently infected with BTV, and was immunologically tolerant to BTV. PMID- 6280529 TI - Effect of bacille Calmette-Guerin immunotherapy on feline sarcoma virus-induced neoplasms in the cat. AB - Kittens infected experimentally with feline sarcoma virus (FeSV) were inoculated with emulsions of bacille Calmette-Guerin (BCG) cell wall preparations to determine the effect of BCG immunotherapy on FeSV-induced sarcoma-genesis. The BCG preparations or emulsificant control preparations were administered (i) subcutaneously at the same time and site as was the FeSV inoculation, (ii) at the same site but 1 week after FeSV inoculation, or (iii) with a mixture of viable autochthonous neoplastic cells approximately 35 days after FeSV inoculation. There was no difference in the percentage of kittens that developed neoplasms, the prepatent period for neoplastic development, the percentage of kittens with neoplastic regression, or the survival rate among groups given BCG, emulsificant control, or FeSV alone. Significantly (P less than 0.05) greater dissemination of neoplasms was seen in groups given BCG or emulsificant control preparations, compared with dissemination in groups given FeSV alone. PMID- 6280528 TI - Isolation of viable canine intestinal lymphocytes. AB - A modification of a previously reported enzymatic technique was used to isolate 60% to 90% pure populations of viable lymphocytes from canine small intestinal mucosa. Mitogenesis assays were then simultaneously performed on these and isolated peripheral blood lymphocytes from the same dogs. A technique to isolate intraepithelial lymphocytes was not successful. Intestinal mucosal lymphocytes were simultaneously purified by 2 different techniques (suspensions B and C), which produced cell populations that responded similarly to phytohemagglutinin (PHA), concanavalin A, and pokeweed mitogens. The peripheral blood lymphocytes demonstrated greater response to PHA and concanavalin A mitogens than did the intestinal mucosal lymphocytes (P less than 0.05). Furthermore, peripheral blood lymphocytes had a significantly higher response to PHA than to pokeweed mitogen (P less than 0.05). The impure preparation of intraepithelial lymphocytes (suspension A) did not show any evidence of reactivity to mitogens. Further studies are needed to determine whether the EDTA or collagenase had any effect upon the intestinal mucosal lymphocyte responsiveness. The present study demonstrated that viable mucosal lymphocytes can be isolated from the canine intestine and that they can maintain their responsiveness to mitogens. PMID- 6280530 TI - In vitro evaluation of B-lymphocyte function in turkeys infected with hemorrhagic enteritis virus. AB - Studies were conducted on B-lymphocyte function in turkeys infected with hemorrhagic enteritis (HE) virus. Hemolytic plaque-forming technique was used to detect antibody-forming cells in turkeys. The plaque-forming cell responses in HE virus-infected and noninfected controls were compared. Results of this study indicated a decreased capability of HE virus-infected turkeys to produce antibodies to sheep RBC. The greatest inhibition of antibody-forming cell production was seen in the turkeys 19 days after exposure to the virus. However, after this period, the turkeys gradually recovered their immunocompetence to sheep RBC. PMID- 6280531 TI - Comparison of aqueous porcine ACTH with synthetic ACTH in adrenal stimulation tests of the female dog. AB - The adrenocortical (plasma corticosteroid) responses in female dogs given porcine ACTH in gelatin (1-39 amino acid sequence) and synthetic ACTH (1-24 amino acid sequence) were compared. Sixteen dogs were used. Each dog underwent 4 different ACTH stimulation studies, these being done with a 4- to 8-week interval. The studies in each dog included injections of 2 doses of porcine ACTH--2.2 IU and 4.4 IU/kg of body weight--and of 2 doses of synthetic ACTH--0.25 mg/dog and 0.50 mg/dog. The dogs were arbitrarily allotted to 4 groups, each group being subjected to a given sequence of stimulation studies. The purpose in this project was to determine whether the established methods for synthetic and porcine ACTH stimulation tests had similar results. Statistical analysis of the 4 stimulation methods revealed no significance (P greater than 0.05) in the resting or poststimulation plasma corticosteroid concentrations. Thus, it was concluded that either recommended method using ACTH (porcine ACTH at 2.2 IU/kg or synthetic ACTH at 0.25 mg/dog) causes maximal secretion of adrenocortical reserve. Either ACTH preparation, using the established method, can be used interchangeably. PMID- 6280532 TI - Characterization of parainfluenza type 3 virus isolated from the lung of a lamb with pneumonia. AB - A virus designated DH-1 was isolated from the lung of a lamb that died during the course of an epizootic of acute respiratory tract disease. The virus contained ribonucleic acid and was sensitive to heat (56 C), lipid solvent, and acid (pH 3.0). Inoculated cell cultures absorbed and culture fluids agglutinated guinea pig erythrocytes. Morphogenic and morphologic characteristics of the isolate were compatible with those of the genus Paramyxovirus of the family Paramyxoviridae. Serologic results indicated that the virus was indistinguishable from prototype parainfluenza virus type 3. PMID- 6280533 TI - Effect of antibacterials, anabolic steroid, and diethyldithiocarbamate on induced acute aflatoxicosis in goats. PMID- 6280534 TI - Antibody, immune complexes, and complement activity fluctuations in kittens with experimentally induced feline infectious peritonitis. AB - Humoral changes were studied in 6 specific-pathogen-free kittens during experimental infection with feline infectious peritonitis virus. Although the incubation period and the duration of the disease differed widely, a similar pattern of rectal temperatures, serum complement values, circulating immune complexes, and antibody titers was found for all kittens during the last 15 to 20 days of life. Antibody formation started 8 to 13 days before death and was accompanied by the appearance of circulating immune complexes with subsequent increased concentrations of complement followed by complement depletion. The data are discussed as evidence for an immune complex pathogenesis of feline infectious peritonitis. PMID- 6280535 TI - Comparison of a direct radioimmunoassay with virus isolation in cell culture for detection of pseudorabies virus in tissues of infected swine. AB - A direct radioimmunoassay (DRIA) was developed for detection of pseudorabies virus (PRV) in tissues of pigs with clinical disease. Homogenates of tissues prepared from 289 swine were tested for PRV with the DRIA and virus-isolation technique (VIT). The virus was detected in 53 (18%) of the homogenates by both DRIA and VIT. Fifteen (5%) others were positive for PRV by VIT, but negative by DRIA. There were no samples negative by VIT and positive by DRIA. A total of 221 (77%) pigs with clinical PRV infection were negative in both DRIA and VIT. Although the VIT is more sensitive than DRIA, the DRIA possesses certain advantages which include reduced requirement for aseptic procedures and cell cultures, rapidity, and increased objectivity. Moreover, high levels of confidence could be attributed to DRIA-positive results, since DRIA-positive/VIT negative results were not obtained. PMID- 6280537 TI - Ultrasonically guided aspiration needle biopsy in disease of the chest. AB - Ultrasonically guided aspiration needle biopsy has been used in our department as a diagnostic procedure in chest disease. Twenty patients with chest disease judged to have lesions abutted to the thoracic wall were selected as candidates for this technique, which was a linear electronic array scanner provided with a specially designed guide channel. In all sixteen patients with malignant disease, this method provided biopsies that yielded malignant cells on cytologic examination. The diagnostic accuracy was excellent and no complications were encountered. PMID- 6280536 TI - Marek's disease in turkeys: lack of protection by vaccination. AB - Herpesvirus of turkeys, a highly effective vaccine against Marek's disease (MD) in chickens, was ineffective in protecting turkeys against MD. Another tissue culture attenuated vaccine virus also protected chickens, but not turkeys, from MD. Intact and immunosuppressed turkey poults inoculated with herpesvirus of turkey developed a persistent viremia, but did not have detectable gross or microscopic lesions. PMID- 6280538 TI - Etidronate disodium in the treatment of Paget's disease of bone. AB - Control of Paget's disease of bone has been possible through treatment with agents that decrease bone resorption; calcitonins, diphosphonates, and mithramycin. The pagetic lesion is not, however, cured. Etidronate disodium is one of the diphosphonates. The clinical improvement attained with this drug has to be set against adverse effects, of which pain is probably the most bothersome in practice. Clinical remission can last as long as 2 years after treatment is stopped. PMID- 6280539 TI - Poliomyelitis prevention. Immunization Practices Advisory Committee Centers for Disease Control. AB - This revised Immunization Practices Advisory Committee recommendation on poliomyelitis prevention addresses issues important in poliomyelitis prevention addresses issues important in poliomyelitis control in the United States today. Specifically, situations that constitute increased risk are defined, and alternatives for protection are outlined. Recommendations for immunization of adults are presented, clarifying the role of inactivated polio vaccine in immunizing adults. These recommendations also address the problems of interrupted immunization schedules and completion of primary immunization. Oral polio vaccine remains the vaccine of choice for primary immunization of children. PMID- 6280540 TI - Naloxone, beta endorphins, and high-altitude pulmonary edema. PMID- 6280541 TI - [Dust and noise pollution in the refractory materials industry]. PMID- 6280543 TI - Role of the catabolite activator protein in the expression of the maltose regulon of Escherichia coli. AB - Using malT-lacZ strains deleted for gene crp, we have shown that the expression of malT is controlled by the catabolite activator protein (CAP), the product of gene crp. malT X mutations were obtained which allowed a malT-lacZ hybrid gene to be expressed at a high level even in the absence of CAP. These mutations were shown to be located in or close to the promoter of the malT gene. We transferred the malT X mutation cis to a wild type malT gene. In the resulting strains, the study of the expression of the three operons in absence or presence of CAP led us to the following conclusion. CAP appears to control malPQ expression mainly if not only by regulating the concentration of MalT protein in the cell. On the other hand it controls the two other operons more stringently both by regulating malT expression and by a more direct action probably exerted on the promoters of these operons. PMID- 6280542 TI - Kinetics of induction of the lamB-gene product in Escherichia coli: appearance of the newly synthesized protein in the cell envelope. PMID- 6280544 TI - Pleiotropic expression of catabolic operons in the absence of the cAMP-CAP complex: the case of the maltose regulon. PMID- 6280545 TI - Regulation by ADH and cellular osmolarity of water permeability in frog urinary bladder: a time course study. PMID- 6280546 TI - Evidence for calmodulin dependence of water permeability in toad urinary bladder. PMID- 6280547 TI - Effects of insulin on sodium transport by the amphibian urinary bladder. PMID- 6280548 TI - Hormone-sensitive ion transport systems in erythrocytes as models for epithelial ion pathways. PMID- 6280549 TI - Membrane beta-receptors: interaction with cytoskeleton in chloride secretory systems. PMID- 6280551 TI - Regulation of dome formation in kidney epithelial cell cultures. PMID- 6280550 TI - Stimulation and inhibition of bicarbonate transport in the turtle bladder. PMID- 6280552 TI - Multiple effects of corticoid hormones on the mammalian nephron. PMID- 6280553 TI - Molecular action of 1,25-dihydroxyvitamin D3: new cultured cell models. PMID- 6280554 TI - The phosphatidate-polyphosphoinositide cycle: activation by parathyroid hormone and dibutyryl-cAMP in rabbit kidney cortex. PMID- 6280555 TI - Hormonal control of chloride secretion by canine tracheal epithelium: an electrophysiologic analysis. AB - A principal event in the stimulation of cyclic AMP-induced active C1 secretion across canine tracheal epithelium is an increase in the C1 permeability of the apical cell membrane. This permits C1 to leave the cell down its electrochemical potential difference across this barrier. C1 transport across the basolateral membrane appears to be nonconductive and may be mediated by NaC1 cotransport. Increased C1 entry across the basolateral membrane would enhance Na entry into the cell and secondarily stimulate Na extrusion from cell to submucosal solution. Increased Na-K-pump activity or an increase in the K permeability of the basolateral membrane may be responsible for the (Formula in text) reduction in basolateral membrane resistance that accompanies stimulation of C1 secretion. Analysis of the electromotive force across the apical cell membrane under conditions where C1 secretion is inhibited or stimulated provides reasonable estimates of intracellular Na and C1 activities for these conditions. PMID- 6280556 TI - Localization of the aldosterone response in rabbit urinary bladder by electrophysiological technics. PMID- 6280557 TI - Hormonal control of secretion in shark rectal gland. PMID- 6280558 TI - Transport properties of intestinal basolateral membranes. AB - Techniques for the isolation and study of basolateral membrane vesicles from the intestinal epithelium have afforded new insights into the mechanisms of intestinal absorption. First, we have confirmed the hypothesis that the second stage of glucose transport involves facilitated diffusion. Second, we have shown that the major system for translocation of neutral amino acids across the basolateral membrane is the classical "L" system. Third, we have established that basolateral membranes contain sodium-dependent transport systems that may be useful in the supply of essential amino acids to the epithelium from the blood. And, finally, our studies of the basolateral (Na + K)-ATPase have clarified the role of this enzyme in sodium absorption. PMID- 6280559 TI - Hormone regulation of parietal cell function: gastrin interaction with a specific receptor. PMID- 6280560 TI - Induction of amiloride-sensitive sodium transport in the intestines by adrenal steroids. PMID- 6280561 TI - Nonlinear characteristics of single neurons in the vestibular nuclei. AB - The characteristics of response of 99 vestibular nuclei neurons were investigated in the cat anesthetized with pentobarbital sodium. All neurons responded to stimulation of the horizontal semicircular canals (HSCC) with prolonged 2, 4, 6, or 8 degrees/second2 angular acceleration. Neural response was recorded utilizing the single-neuron recording technique. The vestibular neurons responded nonlinearly to stimulation of the HSCC. Specifically (1) the time required to reach the maximum response level decreased nonlinearly with an increase in stimulus magnitude (SM); (2) the maximum response level increased nonlinearly with the increase in SM: (3) the neural response to excitatory stimulus was not the mirror image of that to an inhibitory stimulus. Several existing mathematical models were simulated by utilization of the IBM Continuous System Modeling Program, account for the nonlinear response of the vestibular nuclei neurons. The nonlinearities observed in the responses may be due to (1) nonlinear behavior of the end organ, (2) the influence of the CNS including the vestibular efferent system, or (3) the inherent neural properties of the vestibular nuclei. PMID- 6280562 TI - Neuronal interaction between ipsilateral medial and lateral vestibular nuclei. PMID- 6280563 TI - Surface ultrastructure and microviscosity of lymphocyte membranes in myasthenia gravis. PMID- 6280564 TI - Recent advances in the molecular mechanisms of human and animal models of myasthenia gravis. AB - The receptor-channel molecule is a dynamic system which exists in multiple conformations and that is the way we should think of it when we study antibody interaction with the molecule. The results presented here suggest that some antibodies may affect receptor function by occupying sites other than the receptor site. Some of these sites may by exposed only in certain conformations, and occupation of some site by antibodies may effect conformational changes. These small but perhaps important differences in cholinergic channel properties of the myasthenic muscle from the normal one are revealed by studying the effect of myasthenic sera on drug interactions with the channel sites. The sera of myasthenics are able to react with certain channel conformations and are able to affect the interaction of channel antagonists such as H12HTX and QNB. The sera appear to act preferentially with the open conformation of the channel. As a consequence of such an effect, important conformational changes of the channel may fail to occur upon activation. PMID- 6280565 TI - Intercostal muscle biopsy studies in myasthenia gravis: clinical correlations and the direct effects of drugs and myasthenic serum. AB - We have found a wide range of mean MEPP amplitude in intercostal muscle biopsies from 43 patients with MG, including several values in the normal range. There was no correlation between MEPP amplitude and the severity of clinical disease as assessed by manual muscle testing or by single-fiber EMG measurements of jitter in arm muscles. Through most of these patients were in a state of clinical remission or marked improvement after treatment with prednisone, we could not attribute the difference between our results and those of others to this factor alone. The application of morphine, meperidine and aminoglycoside antibiotics to intercostal muscle in vitro confirms effects previously demonstrated in rat muscle: (1) At equal therapeutic concentrations, meperidine has greater neuromuscular blocking effects than does morphine, but neither has significant effects at concentrations achieved in the serum clinically. (2) Tobramycin, netilmicin and neomycin have varying severity and sites of action, but their effects are the same in human myasthenic muscle as in normal rat muscle. Bath application of serum from myasthenic patients produces an acute, reversible worsening of neuromuscular blockade in myasthenic muscle. Electrophysiologic measurements in intercostal biopsies from patients with MG can provide information about the basic abnormality of neuromuscular transmission in this disease and can confirm the relevance of studies made in animal muscle. PMID- 6280566 TI - Recently recognized congenital myasthenic syndromes: (a) end-plate acetylcholine (ACh) esterase deficiency (b) putative abnormality of the ACh induced ion channel (c) putative defect of ACh resynthesis or mobilization - clinical features, ultrastructure and cytochemistry. PMID- 6280567 TI - [A case of association between gastric carcinoma and leukemia (author's transl)]. AB - The authors present a case of association between gastric carcinoma and leukemia. After briefly mention of etiopathogenesys, they discuss the difficulties of differential diagnosis between a primitive neoplasia and a secondary infiltration through the gastric walls, caused by leukemia. They then mention the necessity of a surgical intervention in case of doubtful diagnosis, or in case of a pronounced alteration in the intestinal tract. PMID- 6280568 TI - [Cylindromas of the nasal fossae and base of the skull. Four cases (author's transl)]. PMID- 6280569 TI - Experimental viral infections of the inner ear. III. Viremic spread of reovirus to hamster eighth nerve ganglion cells. PMID- 6280570 TI - [Associated Paget's disease of the vulva and of the nipple in the same patient: pathology with ultrastructural study (author's transl)]. AB - A case of Paget's disease of the vulva associated with Paget's disease of the nipple is reported. When the patient was 74-year-old, Paget's disease of the vulva was diagnosed; four years later, Paget's disease of the nipple was detected. Paget's disease of the vulva, although recurrent, is devoid of underlying infiltrating carcinoma. Eccrine sweats glands show different type of lesions consistent with pagetoid extension, in situ carcinoma and epithelial papillary proliferation. Ultrastructural study demonstrates dense granules within Paget's cells. A positive Thiery's cytochemical reaction indicates that these granules contain glycoprotein. Paget's disease of the nipple is associated with an intraductal carcinoma (cribriform type). Some feature of intralobular carcinoma is also observed. No foci of infiltration are found. Oestrogen receptors are detected by the histofluorescence technique of Sin Hang Lee. Few receptors are seen within the intraductal neoplastic cells and within the Paget's cells whereas no receptor at all, are observed within the keratinocytes. Several reports emphasized the relationship between vulvar Paget's disease and infiltrating breast carcinoma. Associated vulvar and nipple Paget's disease is rather rare and was just observed in two cases. These two observations were associated with a mammary infiltrating carcinoma whereas in our case, it remains an in situ carcinoma. PMID- 6280573 TI - Intracellular perfusion. PMID- 6280571 TI - Growth pattern and prognosis in Wilms' tumour. A longitudinal urographic study. PMID- 6280572 TI - Morphology of cutaneous receptors. PMID- 6280575 TI - Squid axon membrane: impedance decrease to voltage clamp. PMID- 6280574 TI - Electroreception. PMID- 6280576 TI - Levels of adenosine 3',5' cyclic monophosphate and guanosine 3',5' cyclic monophosphate in single urine specimens collected from a large population of healthy subjects. AB - The levels of adenosine 3',5' cyclic monophosphate and guanosine 3',5' cyclic monophosphate have been measured in random urine specimens provided by 95 healthy volunteers with a view to assessing the importance of the effects of certain physiological factors on the cyclic nucleotide levels in this type of specimen, and to establish suitable reference ranges for future patient monitoring studies. Of the factors investigated, only the stage of the menstrual cycle was found to have a significant effect; even this variation was still within the overall normal reference range. Measurements of urine cyclic nucleotides were also made in relation to the method of specimen collection, handling, and storage. From these studies it was concluded that if the specimen is placed in a refrigerator within a few hours of collection and then transferred to a deep-freeze within 48 hours, these factors do not present any major problems for the subsequent assay of the cyclic nucleotides. PMID- 6280577 TI - Congenital cytomegalovirus infection. AB - Three cases of overt congenital cytomegalovirus infection, each with a different mode of presentation, are documented. Common features of presentation were hepatosplenomegaly, skin petechiae, neonatal jaundice and gross hepatic dysfunction. One infant presented with hydrops fetalis, a feature not described before. Two infants survived and are at present developing normally. PMID- 6280578 TI - Red cell enzymopathies in the newborn. I. Evaluation of red cell metabolism. AB - In order to evaluate properly red cell metabolic data obtained in newborns with congenital hemolytic disorders, the unique metabolic characteristics and normal developmental changes that occur prenatally and postnatally are presented. The age-dependent red cell glycolytic enzymes (hexokinase, aldolase, pyruvate kinase) and glucose-6-phosphate dehydrogenase and most glycolytic intermediates are elevated at birth and at 11 to 12 months of age, consistent with the presence of a young red cell population the entire first year of life. However, certain red cell enzymes are elevated out of proportion to the age of the red cell population [phosphoglucose isomerase. glyceraldehyde-3-phosphate dehydrogenase, phosphoglycerate kinase (PGK), and enolase (ENO)] whereas others are decreased [phosphofructokinase (PFK), glutathione peroxidase, carbonic anhydrase, and others]. These metabolic characteristics are felt to be unique and representative of "fetal erythropoiesis." Activities of PGK and ENO decrease the PFK increases toward normal adult values beginning at eight to nine weeks of age. The concentration of glucose-6-phosphate steadily increases after birth and peaks at three to four weeks of age, at a time when PFK activity remains relatively unchanged, suggesting a relative block in glycolysis at the PFK step secondary to an enzyme with both decreased activity and altered kinetic properties (a "fetal" isozyme). Thus, evaluation of red cell enzyme and glycolytic intermediate data obtained in the first year of life should be related to the knowledge that a young red cell population is present and the characteristic unique metabolic red cell alterations described in cord blood persist beyond the immediate neonatal period. PMID- 6280580 TI - [Cartilage and vitamin D in vitro (author's transl)]. AB - Interactions between cartilage and vitamin D metabolism were studied in vitro. Chondrocytes were isolated from rabbit growth plate cartilage and cultured. These cells are able to transform 25-hydroxycholecalciferol in 24, 25 dihydroxycholecalciferol. Metabolic transformation is modulated by 1, 25 dihydroxycholecalciferol but not by parathyroid hormone neither by extracellular calcium concentrations. 24, 25-(OH) 2D3 and 1, 25-(OH) 2D3 are active on the cellular metabolism of cultured chondrocytes in a different way : 24, 25-(OH) 2D3 stimulates the proteoglycan synthesis in "mature" chondrocytes 1, 25-(OH) 2D3 increases DNA polymerase activities in chondrocytes during the logarithmic phase of division. Finally, specific sites of nuclear receptors of 24, 25-(OH) 2D3 are present in chondrocytes. PMID- 6280579 TI - [D-glucose and L-leucine-provoked insulin secretion: possible relationship with stereospecific receptors activation or the metabolism of these nutrients (author's transl)]. PMID- 6280581 TI - Studies of the effects of cholecalciferol upon the metabolism of rachitic chick growth cartilage. AB - When a physiological dose of cholecalciferol was given to rachitic chicks, an early response was increased amino acid incorporation into growth cartilage protein, including collagen, but no evidence was found for induction of specific protein synthesis as in the intestine. Increased release into the medium of 45Ca and of hydroxyproline was observed when growth cartilage tissue was incubated in medium containing 1,25-dihydroxycholecalciferol. These studies suggest that the metabolism of cartilage tissue from the zones of proliferation and maturation was more affected than tissue from the zones of hypertrophy and calcification. Cholecalciferol metabolites may therefore have a direct effect upon chondrocytes as well as on bone cells during endochondral bone formation. PMID- 6280582 TI - Experimental models of virus-induced demyelination of the central nervous system. AB - One of the arguments in favor of a viral pathogenesis for multiple sclerosis is the existence of several experimental and natural animal models of virus-induced primary demyelination. This review deals comprehensively with such models. Well known examples of demyelinating viral infections in their natural host are JHM, Theiler, visna, and canine distemper encephalomyelitides. Recent reports of experimental murine infections with pathogens such as vesicular stomatitis, Chandipura, herpes simplex, Venezuelan equine encephalomyelitis, and Semliki Forest viruses are also discussed. The thrust of the review is to include viral models suspected of producing primary demyelination on an immunopathological basis. PMID- 6280583 TI - Selective fiber vulnerability in acute ischemic neuropathy. AB - To study the relationship between fiber size and vulnerability to ischemia, we produced partial infarction of the proximal posterior tibial nerve of Wistar rats by injecting arachidonic acid into the ipsilateral femoral artery. In the resulting lesion degenerating fibers were concentrated in the center of the nerve. Distal to the infarct, surviving myelinated and unmyelinated fibers were evenly distributed throughout both fascicles. Smaller myelinated fibers (less than 6 micrometers in diameter) were more severely depleted than larger ones ( greater than 6 micrometer). Unmyelinated fibers were reduced in number in proportion to or to a greater extent than myelinated fibers. These findings demonstrate that large myelinated fibers are relatively less vulnerable to peripheral nerve ischemia than smaller fibers, and do not support the contention that unmyelinated fibers are inherently resistant to acute ischemia. PMID- 6280584 TI - Rapid improvement of myasthenia gravis after plasma exchange. AB - Neuromuscular transmission and the staircase phenomenon in the adductor pollicis muscle were studied in six patients with myasthenia gravis before and 24 hours after single plasma exchanges given in short series over one to four weeks. An improvement in neuromuscular transmission was observed in all patients but one within 24 hours, suggesting that an immunological block of receptor sites is reversible or that acetylcholine receptors are rapidly resynthesized. In addition to a neuromuscular transmission defect, two of the patients also showed evidence of impaired excitation-contraction coupling with a negative staircase phenomenon and reduced posttetanic twitch potentiation. This condition became normal after plasma exchange, which may indicate that a reversible immunological impairment of the sarcoplasmic reticulum was present. Improvement lasted up to several months. The anti-acetylcholine receptor antibody titers were lowered in response to single plasma exchanges, but actual titer levels showed poor correlation with clinical and electrophysiological changes. PMID- 6280586 TI - Ionic channels in skeletal muscle. PMID- 6280585 TI - Structural bases for metabolic activity. PMID- 6280587 TI - Myocardial membranes: regulation and function of the sodium pump. PMID- 6280588 TI - Phosphorylation of the sarcoplasmic reticulum and sarcolemma. PMID- 6280589 TI - The action of cardiotoxins on cardiac plasma membranes. PMID- 6280590 TI - Adrenergic receptors in the heart. PMID- 6280591 TI - Bacterial chemotaxis. PMID- 6280592 TI - The physiological basis of taxes in Paramecium. PMID- 6280593 TI - Chemotaxis in Dictyostelium. PMID- 6280594 TI - Receptors for peptide hormones: alterations in diseases of humans. PMID- 6280595 TI - Polypeptide and amine hormone regulation of adenylate cyclase. PMID- 6280596 TI - Induction of cell lysis in Escherichia coli: cooperative effect of nocardicin A and mecillinam. AB - Nocardicin A and mecillinam are two beta-lactam antibiotics with poor bacteriolytic activity against Escherichia coli. However, the combined use of these drugs resulted in the induction of a fast lytic response in E. coli cells. For this cooperative effect to take place, the formation of a complex between penicillin-binding protein 2 and mecillinam is apparently necessary. This suggests that penicillin-binding protein 2 might be actively involved in the response of E. coli to bacteriolytic beta-lactam antibiotics. PMID- 6280598 TI - Effect of rifampin, levamisole, and cytoplasmic protein antigen from Mycobacterium microti on 5'-nucleotidase production by guinea pig macrophages. AB - A decline in 5'-nucleotidase production was observed in short-term tissue culture of guinea pig alveolar, peritoneal, splenic, and liver macrophages during exposure to 10(2) microM rifampicin, 1 microM levamisole, or 10 micrograms of cytoplasmic protein antigen extracted from Mycobacterium microti. Liver macrophage 5'-nucleotidase production was more significantly inhibited by the three agents. Cytoplasmic protein antigen from M. microti was the most potent inhibitor of 5'-nucleotidase production. PMID- 6280597 TI - Single-dose pharmacokinetics of ceftriaxone in infants and young children. AB - The pharmacokinetics of ceftriaxone were studied in five infants (7 to 15 months old) and five young children (24 to 70 months old). Both groups received a single 50-mg/kg dose in an intravenous infusion over 5 min. No major pharmacokinetic differences were observed between the two populations. The total (bound plus unbound) plasma concentration-versus-time data could be described in each case by a biexponential equation. Changes in renal clearance indicated time- and dose- dependent pharmacokinetic behavior. The fraction excreted unchanged in the urine (fu) and the biological half-life (t 1/2 (beta)) were, however, dose independent. The average values were 47% for fu (0 to 12 h) and 6.5 for T 1/2 (beta). Weight corrected total systemic clearance was C1TS = 0.71 ml/min per kg; volume of distribution was VD (beta) = 394 mg/kg. The data support intravenous administration of 50 mg of ceftriaxone per kg of body weight every 12 h in assessing its activity against Haemophilus influenzae, Streptococcus pneumoniae, and Neisseria meningitidis in postneonatal-stage pediatric patients. PMID- 6280599 TI - Pharmacokinetics and bacteriological efficacy of cefoperazone, ceftriaxone, and moxalactam in experimental Streptococcus pneumoniae and Haemophilus influenzae meningitis. AB - The pharmacokinetics and bacteriological efficacy of cefoperazone, cefuroxime, ceftriaxone, and moxalactam were evaluated in the experimental rabbit meningitis model of Haemophilus influenzae type b or Streptococcus pneumoniae infection. The cerebrospinal fluid penetration of these beta-lactam antibiotics was from 3 to 14% and was greater in Haemophilus-infected that in pneumococcus-infected animals. With the exception of moxalactam, the antibacterial activity in cerebrospinal fluid and change in concentration of bacteria during therapy with the test drugs were comparable to those of penicillin G in pneumococcal infection. In animals infected with H. influenzae, cefoperazone, moxalactam, and ceftriaxone were as effective as chloramphenicol in reducing the bacterial counts in cerebrospinal fluid. Moxalactam and ceftriaxone produced the largest cerebrospinal fluid bactericidal titers against this beta-lactamase-producing strain of Haemophilus. On the basis of these data, it was concluded that ceftriaxone and cefoperazone were effective against both pathogens in this meningitis model, whereas moxalactam was effective against only Haemophilus, and cefuroxime was effective against only S. pneumoniae. PMID- 6280601 TI - Occurrence of transposable trimethoprim resistance in clinical isolates of Escherichia coli devoid of self-transmissible resistance plasmids. AB - Fifty trimethoprim-resistant clinical isolates of Escherichia coli, devoid of self transmissible trimethoprim resistance plasmids, were examined for the presence of trimethoprim resistance transposons. Trimethoprim resistance was mobilized from 12 strains by transposition onto plasmid RP4. The trimethoprim resistance transposons isolated comprised two groups: those with and without linked streptomycin resistance. PMID- 6280600 TI - Clinical evaluation of cefotaxime for therapy of lower respiratory tract infections. AB - A clinical trial was designed to evaluate the efficacy and safety of cefotaxime, a new semisynthetic, broad-spectrum cephalosporin, in the therapy of community and hospital-acquired pneumonias. Thirty-nine males (mean age, 65 years) were treated for 41 episodes of pneumonia. Only five patient did not have a serious underlying disease; 15 had two or more significant disorders. Sixty-six percent of these pneumonias were due to Streptococcus pneumoniae or Haemophilus influenzae. The minimal inhibitory concentrations for all bacterial isolates ranged from 0.008 to 4 micrograms/ml. Peak serum cefotaxime levels during therapy ranged from 12 to 124 micrograms/ml 1 h after a 1-g dose. Satisfactory bacteriological and clinical responses were observed in 85% of the cases. Four episodes of pulmonary superinfections due to cefotaxime-resistant gram-negative bacilli were noted, each in a patient being mechanically ventilated. Pseudomonas was involved in each of these superinfections, and three were fatal. No serious toxicity or adverse reaction to cefotaxime was seen. The results of this study suggest that cefotaxime is an affective and well-tolerated new cephalosporin antimicrobial agent for the therapy of pneumonia due to susceptible organisms. PMID- 6280602 TI - Comparative in vitro bactericidal activity of cefonicid, ceftizoxime, and penicillin against group B streptococci. AB - The minimal inhibitory and bactericidal concentrations for 108 group B streptococcal strains were determined for two new cephalosporins, cefonicid and ceftizoxime, and compared to those of penicillin G. Penicillin G was the most active inhibitory and bactericidal agent. Ceftizoxime was a significantly more active inhibitory and bactericidal agent than cefonicid (P less than 0.0005, P less than 0.025, respectively. PMID- 6280603 TI - Combined activity of ketoconazole and 5-fluorocytosine on potentially pathogenic yeasts. AB - In vitro studies based on periodic viable count determinations indicated that the two antifungal drugs ketoconazole and 5-fluorocytosine generally were at least additive in their combined effect on various opportunistic yeast pathogens, including some resistant to 5-fluorocytosine. PMID- 6280604 TI - Inhibitory effects of antiherpesviral thymidine analogs against varicella-zoster virus. AB - Thymidine analogs highly active against herpes simplex virus were compared in their inhibitory action against seven strains of varicella-zoster virus by a plaque reduction assay. E-5-Bromovinyl-arabinosyluracil (BV-ara-U) was most active, followed by E-5 chlorovinyl-arabinosyluracil, E-5-bromovinyl-2' deoxyuridine (BV-dUrd), 2'-fluoro-5-methyl-arabinosyluracil, 2'-fluoro-5-iodo arabinosylcytosine, arabinosylthymine, 5-vinyl-arabinosyluracil, acycloguanosine, and 5-iodo-2'-deoxyuridine, in order to decreasing activity. BV-ara-U was more than 10 times as active as BV-dUrd and almost completely inhibited plaque development of five strains of varicella-zoster virus at a concentration as low as 1 ng/ml. PMID- 6280605 TI - Acyclovir - a review of the preclinical and early clinical data of a new antiherpes drug. AB - A review of the literature on acyclovir (Zovirax), a new antiherpes drug, with particular activity against herpes simplex virus types I and II and also against varicella-zoster, Epstein-Barr, cytomegalo and herpes B viruses, is presented. The article deals with 'in vitro' and 'in vivo' efficacy in animals, animal toxicity, latency and resistance, the mechanism of action and early clinical experience. PMID- 6280606 TI - Antiherpes activity of [E]-5-(1-propenyl)-2'-deoxyuridine and 5-(1-propenyl)-1 beta-D-arabinofuranosyluracil. AB - 5-(1-Propenyl)-1-beta-D-arabinofuranosyluracil has been synthesized, and this compound and [E]-5-(-propenyl)-2'-deoxyuridine have been tested for inhibition of herpes virus multiplication. Only [E]-5-(1-propenyl)-2'-deoxyuridine was found to be an active inhibitor reducing by 50% the plaque formation of herpes simplex virus type 1 (HSV-1) at about 1 muM. A comparison to the bromovinyl derivatives showed the following order of descending activity; [E]-5-(2-bromovinyl)-2' deoxyuridine greater than 5-(2-bromovinyl)-1-beta-D-arabinofuranosyluracil greater than or equal to [E]-5-(1-propenyl)-2'-deoxyuridine greater than 5-(1 propenyl)-1-beta-arabinofuranosyluracil. HSV-1 mutants lacking thymidine kinase or resistant against acycloguanosine were resistant against [E]-5-(1-propenyl)-2' deoxyuridine. All compounds seemed to be phosphorylated by HSV-1 thymidine kinase in a cell-free assay. The compounds were phosphorylated to a lower extent by cellular or HSV-2 thymidine kinase, and the HSV-2 strains tested were inhibited by less than 50% at 100 muM in plaque assays. A selective inhibition of HAV-1 DNA synthesis by [E]-5-(1-propenyl)-2'-deoxyuridine was observed in infected cells indicating an effect on viral DNA polymerase. [E]-5-(1-Propenyl)-2'-deoxyuridine had a low cellular toxicity and a therapeutic effect when applied topically to HSV-1-infected guinea pig skin. PMID- 6280607 TI - A comparison of phosphonoacetic acid and phosphonoformic acid activity in genital herpes simplex virus type 1 and type 2 infections of mice. AB - The activity of phosphonoacetic acid (PAA) and phosphonoformic acid (PFA) against four strains of herpes simplex virus type 1 (HSV-1) and four strains of HSV-2 were compared in tissue culture and in a murine model of genital herpes. In mouse embryo fibroblast cells, both drugs were three-fold more active against the HSV-1 strains than against the HSV-2 strains. In contrast, in the animal model infections, PAA appeared to be more active against the HSV-2 strains, while PFA was equally effective against both HSV types. In mice infected intravaginally with HSV-2 and treated with intravaginal 5% PAA, none of the treated mice became infected, replication of virus in the genital tract was completely inhibited, none of the infected mice died from encephalitis, and latent infection in lumbosacral ganglia of surviving animals was completely prevented. In HSV-1 genital infection treated with PAA, 20-60% of mice became infected, replication of virus in the genital tract was strikingly reduced, none of the infected mice died, and latent infection was completely prevented. In both HSV-2 and HSV-1 genital infections, 20-70% of animals treated with 8% PFA became infected, growth of virus in the genital tract was reduced significantly but not completely suppressed, mortality was variably altered, and there was a trend towards reduction in the frequently of latent infection. These results indicate that HSV 1 strains are more sensitive to PAA and PFA in tissue culture, but the HSV-2 strains are generally more amenable to therapy in the murine model of genital herpes. Although PAA appeared to be more active that PFA in the genital infection, both drugs significantly altered the course of the infection. Since dermal toxicity associated with PAA precludes its use in humans and since PFA is already undergoing trials in patients with recurrent herpes labialis, the current results suggest that topical PFA deserved further evaluation in the treatment of mucocutaneous HSV infections, including genital herpes. PMID- 6280608 TI - Antiviral effects of single-stranded polynucleotide inhibitors of the influenza virion-associated transcriptase against influenza virus infection of hamsters and ferrets. AB - Administration of a single-stranded polynucleotide copolymer containing 9% cytidine residues and 91% 4-thiouridine residues [poly(C,S4U10)], a known potent inhibitor of the virion transcriptase of influenza viruses, suppressed the amount of virus recoverable from the nasal washes of influenza virus-infected hamsters and ferrets. The incidence of sneezing and nasal discharge in infected ferrets was also reduced. In hamsters, poly(C,S4U10) was more effective than amantadine HCl or Virazole. Polyinosinic acid in combination with poly-5-hydroxy cytidylic acid also had anti-influenza effects. Poly(C,S4U10) annealed to polyadenylic acid was not effective, nor was the double-stranded polymer (polyinosinic acid) . (polycytidylic acid) even when complexed with carboxymethylcellulose and polylysine. No toxic effects of poly(C,S4U10) were apparent in the treated hamsters and ferrets, and high doses (greater than or equal to 2.86 g/kg) administered intraperitoneally to mice produced no adverse effects. PMID- 6280609 TI - Development of age-dependent resistance to sindbis virus encephalitis: correlation with inactivation of virus within the blood stream. AB - The clearance of Sindbis virus from the blood stream and its localization in the reticuloendothelial system (RES) was studied in mice susceptible (2 weeks old) and resistant (6 weeks old) to fatal Sindbis virus encephalitis. No significant differences in the relative capacity of 2-week-old and 6-week-old mice to remove 125I-labelled virus from the blood stream and to localize virus within the liver were observed. However, the decline of infectious virus was more rapid in the blood of adult mice. These studies suggest that, in addition to physical removal of virus by the RES, inactivation of virus in the blood serum stream plays an important role in limiting viremia during infections with Sindbis virus. PMID- 6280610 TI - Inhibitory activity of the new adamantane derivative N,N'-bis(ethylene)-P(1 adamantyl)-phosphonic diamide against Rous sarcoma virus. AB - A new adamantane derivative N,N'-bis (ethylene)P(1-adamantyl)-phosphonis diamide, was found to inhibit Rous sarcoma virus replication in much the same manner as reported for the parent compound, 1-adamantanamine hydrochloride. PMID- 6280611 TI - Ozone inactivation of cell-associated viruses. AB - The inactivation of HEp-2 cell-associated poliovirus (Sabin 1) and coxsackievirus A9 was investigated in three experimental systems, using ozone as a disinfectant. The cell-associated viral samples were adjusted to a turbidity of 5 nephelometric turbidity units. The cell-associated poliovirus and coxsackievirus samples demonstrated survival in a continuous-flow ozonation system at applied ozone dosages of 4.06 and 4.68 mg/liter, respectively, for 30 s. Unassociated viral controls were inactivated by the application of 0.081 mg of ozone per liter for 10 s. Ultrasonic treatment of cell-associated enteric viruses did not increase inactivation of the cell-associated viruses. The batch reactor with a declining ozone residual did not effect total inactivation of either cell-associated enteric virus. These cell-associated viruses were completely inactivated after exposure to ozone in a batch reactor using continuous ozonation. Inactivation of cell-associated poliovirus required a 2-min contact period with an applied ozone dosage of 6.82 mg/liter and a residual ozone concentration of 4.70 mg/liter, whereas the coxsackievirus was completely inactivated after a 5-min exposure to an applied ozone dosage of 4.81 mg/liter with an ozone residual of 2.18 mg/liter. These data indicate that viruses associated with cells or cell fragments are protected from inactivation by ozone concentrations that readily inactivate purified virus. The cell-associated viral samples used in this research contained particles that were 10 to 15 microns in size. Use of a filtration system before ozonation would remove these particles, thereby facilitating inactivation of any remaining viruses associated with cellular fragments. PMID- 6280612 TI - The effect of diabetes on protein synthesis and the respiratory chain of rat skeletal muscle and kidney mitochondria. PMID- 6280613 TI - A reaction of the superoxide radical with tetrapyrroles. PMID- 6280614 TI - Physiochemical properties of ferralterin. A regulatory iron-sulfur protein functional in oxygenic photosynthesis. PMID- 6280616 TI - Stimulation of albumin synthesis in mouse hepatoma cells by Bt2cAMP: cell cycle specificity. PMID- 6280615 TI - Adenylate cyclase activation by GTP analogs. PMID- 6280617 TI - Differences between CTP and ATP as substrates for the (Na + K)-ATPase. PMID- 6280618 TI - Conversion of adrenergic regulation of glycogen phosphorylase and synthase from an alpha to a beta type during primary culture of rat hepatocytes. PMID- 6280619 TI - Occurrence of a new corticotropin-like intermediate lobe peptide in salmon pituitary glands. PMID- 6280620 TI - Inactivation of fructose 1,6-bisphosphatase by a lysosomal proteinase is reversed by cystamine. PMID- 6280621 TI - Whole foods and increased dietary fibre improve blood glucose control in diabetic children. AB - In prescribing a diabetic diet more attention has traditionally been paid to the amount of dietary carbohydrate than to its type or structure. We have compared the effect on blood glucose of substituting unrefined, whole foods for refined, processed foods in liberal carbohydrate diets (50-55% of dietary energy) eaten by 10 diabetic children in a randomised crossover study. All measurements were made at home. The unrefined diet used whole foods (including) dried beans) supplying 60 g/day of dietary fibre. The refined diet used processed foods supplying 20 g/day of dietary fibre. Diets were isocaloric for carbohydrate, fat, and protein. Glycaemic control was assessed by daily urine analysis for glucose, home blood glucose measurements, glycosylated haemoglobin, and by a 24-hour profile of blood and urinary glucose carried out at home after 6 weeks on each diet. Glycaemic control was significantly better on the unrefined diet. On profile days mean blood glucose levels on the unrefined and refined diets respectively were: preprandial: 5.5 and 8.4 mmol/l; postprandial 8.5 and 12.2 mmol/l. The mean 24 hour urinary glucose excretion on the unrefined diet was 9.3 g and on the refined diet was 38.0 g. Six months after the study the children were eating appreciably more dietary fibre than before (mean increase 13.6 g/day). Attention to food type and structure can improve blood glucose control in diabetic children and should provide an acceptable and more rational basis for dietary prescription than one based on carbohydrate quantity alone. PMID- 6280622 TI - Renal threshold phosphate concentration (TmPO4/GFR). AB - The ratio of maximum rate of renal tubular reabsorption of phosphate to glomerular filtration rate (TmPO4/GFR) was determined in 546 schoolchildren, aged between 6 and 17.9 years, using the nomogram of Walton and Bijvoet.1 TmPO4/GFR correlated with chronological age in girls and boys and in each remained significantly higher than in adults. TmPO4/GFR in the children correlated neither with fasting serum immunoreactive calcitonin and parathyroid hormone levels nor with the urinary cyclic AMP excretion. The study showed a parallel decrease in TmPO4/GFR, excretion of total hydroxyproline and serum alkaline phosphatase activities after puberty, with a significant relationship of both these indices of bone turnover to TmPO4/GFR values. This indicates that the high renal phosphate threshold of children may be an important factor for bone mineralisation by providing high extracellular inorganic phosphate concentrations during normal growth. PMID- 6280623 TI - Oral converting enzyme inhibitor in malignant hypertension. AB - Malignant hypertension, which developed in a 9-year-old boy after an episode of haemolytic uraemic syndrome, could not be controlled with antihypertensive agents. However, treatment with oral converting enzyme inhibitor (captopril) was effective in controlling the blood pressure and it averted bilateral nephrectomy. No adverse effects from the drug were noted. PMID- 6280624 TI - [Rotaviruses as causal agents of acute gastroenteritis. Virological and serological studies in samples of the population of Palermo]. PMID- 6280625 TI - Age changes in rat testicular capsular and parenchymal delta 13-reductase and 15 hydroxyprostaglandin dehydrogenase activities. AB - Prostaglandin dehydrogenase (PGDH), delta 13-reductase, and total PGDH activities (the sum of the first two activities) were measured in rat testicular parenchymal and capsular preparations from 11 through 81 days of age. delta-13 Reductase activity closely paralleled PGDH activity and was substrate dependent except for the parenchymal activity at 61 and 81 days of age when something appeared to enhance its activity. All three activities, when expressed on a per milligram of tissue basis, were elevated at 11 days postpartum and then decreased to a low value at 21 days of age for the parenchyma and 21-32 days of age for the capsule. The elevated activity at 11 days of age suggested a possible elevation of enzyme activity by placental gonadotropins or possibly progesterone. Maximal, activity was observed at 51 days of age for the parenchyma and 61 days for the capsules with diminished activities observed with advancing age except for the parenchymal delta 13-reductase activity. On a per milligram of tissue basis, the capsule demonstrated more enzyme activity than did the parenchyma with maximal activities being observed at 51 days of age. Possible control mechanisms were the following: substrate induction, gonadotropins, testosterone, or those factors regulating testosterone secretion. PMID- 6280626 TI - The decrease of neuromuscular transmission by adenosine depends on previous neuromuscular depression. AB - The action of adenosine (0.5 mM) on neuromuscular transmission has been studied in the presence of progressively blocking concentrations of tubocurarine, tetrodotoxin, and high Mg2+/low Ca2+ solutions. Adenosine enhanced the inhibitory effects elicited either by tubocurarine (4 X 10(-7) - 12 X 10(-7) M), high Mg2+(5 - 11 mM) or low CA2+(0.8 - 0.3 mM) concentrations. The effect of adenosine depended upon the size of the previous neuromuscular inhibition. In the presence of tubocurarine, the inhibitory effects of adenosine were greater at room temperature than at 37 degrees C; however, adenosine was more potent at 37 degrees C than at room temperature in solutions with high Mg2+ or low Ca2+ concentrations. Adenosine did not change the inhibitory effects of tetrodotoxin either at 37 degrees C or at room temperature. These findings suggest that the partial neuromuscular blockade, for the inhibitory action of adenosine to be apparent, has to take place either postsynaptically via the blockade of ACh receptors or presynaptically at the nerve endings through a decrease in the evoked output of the transmitter. Since ATP and/or adenosine are released at the neuromuscular junction, a potential involvement of these substances in conditions with impairment of neuromuscular transmission (e.g. myasthenia gravis) is discussed. PMID- 6280627 TI - The involvement of catecholaminergic mechanisms in the appearance of wet dog shakes produced by carbachol chloride in rats. AB - Noradrenaline (NA), methoxamine, dopamine (DA), given intracerebroventricularly (ICV), and L-DOPA, administered systemically, significantly blocked wet dog shakes (WDS) produced by carbachol chloride (10 microgram/10 microliter, ICV) in rats. Reserpine, alpha-methyl-p-tyrosine and FLA 63 did not affect WDS, while diethyldithiocarbamic acid depressed it. Aceperone and yohimbine weakened shaking response to carbachol but phentolamine given ICV showed no effect on WDS. Propranolol and isoproterenol administered ICV did not significantly influence WDS. Apomorphine failed to affect WDS induced by carbachol. Pimozide and spiperone were also ineffective against WDS, but amphetamine and metoclopramide efficiently blocked it. Selective depletion of brain NA concentration considerably enhanced WDS, while selective depletion of brain DA concentration failed to affect it. These results suggest that carbachol-induced WDS behavior is under the inhibitory control of noradrenergic neurons. PMID- 6280628 TI - Effects of butoprozine on ionic currents in frog atrial and ferret ventricular fibres. Comparison with amiodarone and verapamil. AB - The effects of butoprozine on cardiac membrane currents of frog atrial and ferret ventricular fibres were investigated in voltage clamp conditions by means of the double sucrose gap technique. Comparison was made with amiodarone and verapamil. Butoprozine was found to decrease the fast sodium and slow calcium inward currents and the delayed outward current in a dose-dependent manner. The reactivation kinetics of both inward currents were reduced by butoprozine. Amiodarone principally decreased the delayed outward current but high concentrations also depressed the inward currents. In the same way, small concentrations of verapamil inhibited the slow inward calcium current while high concentrations also affected the delayed outward and the fast inward sodium currents. The reactivation kinetics of both inward currents were reduced by amiodarone. These of the fast inward current remained unaffected by verapamil while these of the slow inward current were reduced or sometimes not appreciably changed. The effects of butoprozine on the ionic currents accounted for the modifications observed on the action potential and automaticity and this study allowed the understanding of the mechanisms by which the drug can exert its antiarrhythmic action. Some of them are common to amiodarone and verapamil. PMID- 6280629 TI - Hypotensive properties of benzodioxane derivatives structurally related to R 28935. Comparison of activity with some receptor affinities. AB - Hypotensive activities were determined of some derivatives of erythro 1-(1-[2 (1,4-benzodioxane-2-yl)-2-hydroxyethyl]-4-piperidyl)-2-benzimidazolinone (R 28935) following intravenous administration to anaesthetized normotensive rats. Introduction of chlorine into the benzimidazolinone part of R 28935 negatively influenced the hypotensive potency as did opening of the dioxane ring. An appropriate substituent restored the hypotensive effectiveness of the "opened" structures. In general, the compounds were weak inhibitors of the specific binding of (3H)-prazosin (alpha 1-adrenoceptors), (3H)-clonidine (alpha 2 adrenoceptors) and (3H)-dihydromorphine (opiate-receptors) to rat brain membranes. The relative order of affinity for either receptor did not correspond with that of the hypotensive activity. Previous (-15 min) intravenous treatment with phentolamine (0.2 mg/kg) abolished the depressor effect of prazosin and diminished that of the threo form R 29814 as well as of all "opened" congeners, but did not significantly reduce the hypotensive response to R 28935 and the other erythro structures. It is concluded that neither alpha 1- and alpha 2- nor opiate-receptors are involved as the primary targets for R 28935 and its congeneric drugs to induce hypotension. The exact mechanism of action remains therefore unsolved. Erythro R 28935 and the other racemic erythro mixtures are centrally acting hypotensive agents. A peripheral alpha-sympatholytic component may contribute to the overall depressor effect of threo R 29814 and the "opened" derivatives. PMID- 6280630 TI - Chemotherapy of germ cell tumors in women. PMID- 6280631 TI - [Cellular effects of anti-arrhythmia drugs. Their predictive value of therapeutic effects]. PMID- 6280632 TI - [Diagnosis of myocardial infarct after cardiac surgery using technetium scintigraphy. Apropos of 106 cases]. PMID- 6280633 TI - [Use of a simplified exercise test in the study of a beta-blocker]. PMID- 6280635 TI - [The role of mammography in the early detection of breast cancer (author's transl)]. PMID- 6280637 TI - Hepatocellular carcinoma metastatic to the choroid. PMID- 6280634 TI - Electron transport-linked proton translocation at nitrite reduction in Campylobacter sputorum subspecies bubulus. AB - Campylobacter sputorum subspecies bubulus contains a membrane-bound nitrite reductase which catalyses the six-electron reduction of nitrite to ammonia. Formate and L-lactate are used as hydrogen donors. Cells of C. sputorum grown with nitrate or nitrite contain cytochromes of the b- and c-type and a carbon monoxide-binding cytochrome c. In addition, a special membrane-bound carbon monoxide-binding pigment is found. Nitrite reduction with formate or L-lactate as a hydrogen donor is strongly inhibited by 2-n-heptyl-4-hydroxyquinoline-N-oxide (HQNO). Nitrite reduction by bacterial suspensions with lactate as a hydrogen donor is strongly inhibited by carbonylcyanide-m-chlorophenylhydrazone (CCCP) whereas nitrite reduction with formate as a hydrogen donor is not inhibited at all. Leads to H+/O values and leads to H+/NO-2 values were measured with ascorbate + N,N,N',N'-tetramethyl-p-phenylenediamine (TMPD), formate (in the absence and presence of carbonic anhydrase) and L-lactate as a hydrogen donor. The results are summarized in a scheme for electron transport from formate or lactate to oxygen or nitrite which shows a periplasmic orientation of formate dehydrogenase and nitrite reductase and a cytoplasmic orientation of lactate dehydrogenase and oxygen reduction, and which shows proton translocation with a leads to H+/2e value of 2.0. The leads to H+/O and leads to H+/NO-2 values predicted by this scheme are in good agreement with the experimental values. PMID- 6280636 TI - Ribosome-lamellae complexes in a symptomatic insulinoma. An ultrastructural study. AB - Ribosome-lamellae complexes (RLCs) were identified in approximately 20% of the tumor cells from a functional insulinoma studied by electron microscopy. Whereas these peculiar cytoplasmic inclusions are commonly seen in hairy cell leukemia (HCL) and can be present in a few cells in other hematologic disorders, to our knowledge they have never been documented in insulinoma. In fact, they are exceedingly rare in epithelial tumors. A review of the literature uncovered only two instances, one in an adenoma of the adrenal cortex, and one in a paraganglioma. In these two cases and in the insulinoma reported here, RPCs tend to be longer, have fewer lamellae, and occur in greater numbers per single cell than in HCL. Although the nature and significance of RLCs remain poorly understood, the fact that all three epithelial neoplasms in which they have been observed belong to the endocrine system is noteworthy. PMID- 6280638 TI - The detection of the factor lowering the immunogenicity of toxoid of Clostridium perfringens type A. PMID- 6280639 TI - Comparative electron microscopic study of an imported pathogenic strain and an autochthonous nonpathogenic strain of E. histolytica with particular reference of nuclear inclusions. PMID- 6280640 TI - [Report of 2 cases of chemodectoma of the carotid glomus]. PMID- 6280641 TI - [Tuberculin intradermoreaction as a survival test in patients with inoperable pulmonary carcinoma]. PMID- 6280642 TI - [Blood levels of alpha 1- antitrypsin in pulmonary neoplasms]. PMID- 6280643 TI - [Hepatitis in infectious mononucleosis]. PMID- 6280644 TI - [Vesicular mole. Epidemiological considerations]. AB - A full and detailed review of trophoblast disease at Rome University's Obstetrics and Gynecology Clinic led to a re-examination of some of the classical epidemiological factors in this pathology: age, parity and blood group. The precise incidence of this condition is difficult to establish from hospital series alone. In spite of numerous clinical studies, its aetiology has not yet been fully clarified. The various environmental socio-economic, racial, genetic and other factors, which are sometimes quoted, are too heterogeneous and contradictory to provide significant results. Of evaluated risk parameters, only pregnancy at an advanced age appears to carry increased risk. For these reasons, and given the low incidence of trophoblast pathology, it is recommended that a register of the disease be instituted, at least at Regional level. PMID- 6280645 TI - An introduction to radionuclide ventriculography. PMID- 6280646 TI - [Glomangioma of the stomach (pathology, pathogenesis and classification)]. AB - Twenty-two glomus tumors of the stomach were studied. Their mandatory components were shown to include vascular structures, elongated cells similar to smooth muscle elements, and the so-called epithelioid cells with clear perinuclear zones of artificial nature. Among gastric glomus tumors, mature, growing and malignant forms should be distinguished. The results of electron microscopic examinations suggest the smooth muscle origin of the glomus neoplasias. PMID- 6280647 TI - [Role of atypical sex cells in the histogenesis of germinative testicular tumors]. AB - Histochemical and electron microscopic study of 23 germinative tumors of the testicle and organ tissue beyond the tumor was carried out. In 78% of the observations, normal seminiferous tubules were found to contain a special kind of cell elements, atypical sex cells (ASC), not observed in health. Ultrastructural and histochemical features of undifferentiated cells comprising seminoma and embryonal cancer were found to be similar to those of ASC and gonocytes of human embryo. The results suggest that ASC are the stem tumor cells for human germinative neoplasia which are characterized by single cytohistogenesis. PMID- 6280648 TI - [Heterotopic mesodermal tumor of the fallopian tube]. AB - An observation of heterologous mesodermal tumor of the fallopian tube in a woman of 53 is described. The tumor had the structure of adenoacanthocarcinosarcoma with inclusions of atypical cartilage. The patient died 6 months after the operation from generalized process. The rare localization of this tumor is noted. PMID- 6280649 TI - [Current theories on the nature of myoepithelial cells and their role in the morphogenesis of dyshormonal hyperplasia and breast neoplasms]. AB - An analysis of current concepts on the origin and potentials of neoplastic transformation of myoepithelial cells (MC) of the mammary glands is presented. The epithelial and smooth-muscle nature of these cells is discussed. The hypothesis of mixed (epithelial and myoepithelial) nature of mammary gland carcinoma and possibilities of neoplastic transformation of poorly-differentiated forms of MC are considered. Methods for identification of cells of the myoepithelial origin are described. PMID- 6280650 TI - [Edematous form of neurocysticercosis: report of 4 cases]. AB - Four cases of the oedematous form of neurocysticercosis are reported. This form is relatively rare and characterized by symptoms of intracranial hypertension and severe bilateral papilledema. Focal convulsive disorders are common. CSF studies show positive reaction for cysticercosis and ventriculography, the presence of small, narrowed lateral ventricles. Treatment based on ACTH and dexamethasone revelead good evolution as far as intracranial hypertension is concerned, not requiring surgical decompression as used before. PMID- 6280651 TI - [Use of cerebral cortex gangliosides in peripheral neuropathies]. AB - Personal experience with brain gangliosides in the treatment of peripheral neuropathies is reported. The clinical and electromyographic trials were efficacious in 30 of 40 treated cases. The authors emphasize their better results in peripheral facial palsy cases. PMID- 6280652 TI - Late-life chronic peripheral neuropathy of obscure nature. PMID- 6280653 TI - Growth hormone responses to the stimulation of hypothalamic glucoreceptors in diabetic retinopathy. Effects of catecholamine precursors and dopamine antagonists. AB - The effects of the stimulation of glucoreceptors located at the level of the lateral hypothalamic nuclei and sensitive to intracellular glucose deprivation on growth (GH) release are well documented in the monkey. In this study the effect of a reproducible hypoglycemic stimulatory procedure was controlled by continuously monitoring blood glucose (BG) levels in normal subjects and in a series of 120 diabetic patients. In contrast to control subjects, GH responses to similar hypoglycemic nadirs were not reproducible in diabetics (with or without diabetic retinopathy). The infusion of levodopa, precursor of both norepinephrine (NE) and dopamine, during BG decline rendered the GH response to hypoglycemia consistently reproducible. To investigate the site of action of levodopa, a group of diabetics were pretreated with a centrally active dopamine antagonist, metoclopramide hydrochloride. The pattern of GH release was similar in both groups. These data suggest that in diabetes the interaction of levodopa and adequate stimulation of hypothalamic glucoreceptor systems is mediated by an NE transmission. PMID- 6280654 TI - Developmental changes in the collagens and some collagenolytic activities in bovine dental pulps. AB - Collagen concentration was fairly low in young bovine 3rd premolar pulps, about 9 per cent by dry weight of pulp in stage I, but it increased consistently up to more than 25 per cent at stage V as the pulp matured. The ratio of type III to type I collagen similarly increased from 13 per cent at stage I to 32 per cent at stage V. Both collagenase and possibly collagenolytic neutral peptidases showed significantly high activity in the early stage of pulp development where the rate of collagen synthesis was also elevated. Higher rates of both collagen synthesis and its degradation might explain the low concentration and content in the early stage of pulp development. PMID- 6280655 TI - Ultrastructural phosphatase cytochemistry of the intercalary ducts of the parotid and submandibular salivary glands of man. AB - Cells of the intercalary ducts showed concentrations of secretory granules adjacent to the luminal plasma membrane. Evidence of thiamine-pyrophosphatase activity was seen in the Golgi apparatus and of acid-phosphatase activity in GERL like structure, lysosomes and immature secretory granules. Adenosine triphosphatase reaction-product was present along surfaces of myoepithelial cells and to a lesser extent along contiguous surfaces of duct cells. The findings indicate secretory activity in the intercalary duct cells. PMID- 6280656 TI - Influence of thyroid hormone on (Na+ + K+)-dependent adenosine triphosphatase activity in rat submandibular glands. AB - Surgical thyroidectomy decreased (Na+ + K+)-dependent adenosine triphosphatase (Na-K-ATPase) activity in rat submandibular glands. Three successive doses of triiodothyronine (100 microgram/100 g of body wt) to euthyroid and hypothyroid rats produced 24 and 23 per cent increase in the enzyme activity, compared to control values. When hypothyroid rats were given smaller doses of triiodothyronine (4 microgram/100 g body wt every 48 h) for 27 days, Na-K-ATPase activity increased 40 per cent over the corresponding values in the control hypothyroid rats. Thus, thyroid hormone acts on rat submandibular glands to increase the Na-K-ATPase activity of that target tissue. PMID- 6280657 TI - Intra-arterial cisplatin treatment of adenoid cystic carcinoma. AB - Adenoid cystic carcinoma (ACC) is unique among salivary gland tumors in both its natural history and in its response to nonsurgical treatment methods (ie, radiation and chemotherapy). The chemotherapeutic agent, cisplatin, seems to be unique in its ability to affect ACC. It might be that ACCs of the minor salivary glands are especially vulnerable to the intra-arterial method of administering this drug, because there are accessible feeding vessels to most locations in which these tumors occur. We have treated four patients with advanced ACC with intra-arterial cisplatin. The responses suggest that this method may be a useful adjunct in the management of this tumor both as a preoperative and as a palliative measure. PMID- 6280658 TI - Epstein-Barr virus and a unilateral neck mass. AB - Although Epstein-Barr infectious mononucleosis (IM) might be considered in the differential diagnosis of multiple neck masses in a young adult, single large masses are not usually associated with IM. This patient did have a solitary mass. Despite lacking other signs of IM, he did have a positive heterophil agglutination titer and reactive lymphocytes. Other serologic tests can be of value in confirming the diagnosis, especially if the heterophil titer and reactive lymphocytes are absent. These tests are viral capsid antigen IgM and IgG, early antigen, and nuclear antigen. Surgeons should be aware of their usefulness, especially when trying to confirm a difficult diagnosis. PMID- 6280659 TI - Silica bonded investment and diatomite. PMID- 6280660 TI - Herpes zoster. The gp view. AB - Herpes zoster is a fascinating disease with many lessons for the general practitioner. This article raises some practical questions about its diagnosis and management. PMID- 6280661 TI - Herpes zoster: complications pathogenesis and pathology. The specialist view. AB - The term herpes zoster is derived from the Greek herpes: to creep and zoster: a belt or girdle. Shingles is from the Latin cingere: to gird, which was corrupted to mean a belt of girdle for the human form. A typical attack of herpes zoster is usually not difficult to recognise, but it is important to be aware of uncommon manifestations and complications. PMID- 6280662 TI - Agonist-stimulated breakdown of myo-[2-3H] inositol-labelled phosphatidylinositol in mouse pancreas. AB - Agonist-stimulated hydrolysis of phosphatidylinositol (PI) in cell membranes has been proposed to lead to an increase of cytosol calcium concentration and activation of the cellular response in certain smooth muscles and glands. A method is described which allows the rapid, reproducible measurement of hydrolysis of phosphatidylinositol in mouse pancreas. The technique involves the in vivo labelling of the pancreas with myo-[2-3H] inositol. The majority of the label incorporated into phospholipids is in the form of PI, with only a small proportion of label in di- and tri-phosphoinositides. Tissue pieces of the labelled pancreata are incubated in vitro in the presence or absence of secretagogues, and the PI in homogenates of these pieces is precipitated with trichloroacetic acid. No PI remains in the acid-soluble supernatant. This technique does not require the time-consuming extraction and chromatographic separation of lipids which has been necessary in other assays of PI hydrolysis. Using this method, we have confirmed that PI breakdown is stimulated by carbachol and cholecystokininoctapeptide in the presence or absence of extracellular Ca2+. Agonist-stimulated hydrolysis of PI was potentiated by extracellular Ca2+, and was not dependent on agonist-activated Na+ influx. This technique will facilitate the investigation of the importance of PI breakdown in stimulus-response coupling. PMID- 6280663 TI - The presence of oncofoetal antigens in large bowel carcinoma. AB - This paper is an immunohistological study of the occurrence of the oncofoetal antigens, (carcinoembryonic antigen (CEA), small intestine mucin antigen (SIMA), and normal large bowel mucin antigen (LIMA) in 60 surgically resected colons: 10 non-malignant specimens and 50 colorectal carcinomas. SIMA is a new oncofoetal antigen found in mucinous carcinoma of the large bowel. In the adult it is normally present only in the duodenum and jejunum. Of the 50 carcinoma specimens, 13 were mucinous, 17 non-mucinous and 20 mixed mucinous and non-mucinous. LIMA was the only antigen detected in the mucosa of non-malignant specimens. In mucinous carcinomas only SIMA was present, whilst in the non-mucinous specimens CEA was always found and to a lesser extent LIMA. The same relationship was observed in mixed tumours: SIMA in mucinous and CEA-LIMA in the non-mucinous parts. In the mucosa adjacent to the cancer in all 50 cases there was evidence of an increase or decrease in LIMA. In 42 cases (84%) both oncofoetal antigens (CEA and SIMA) could also be detected in this transitional or perineoplastic epithelium at varying distances from the tumour. These results provide evidence to suggest that the majority of large bowel carcinomas occur in areas of metaplastic change. PMID- 6280664 TI - Local recurrence of breast cancer: its significance in relation to breast reconstruction. AB - Questions regarding local recurrences are a major concern in considering mastectomy patients for breast reconstruction. A critical analysis of the literature on local recurrences indirectly provides answers to several of these concerns. Local recurrence is a relatively predictable event, and is uncommon in those with a favourable breast cancer. Its presence reflects concurrent systemic disease which is usually lethal within a short time. Concern over subsequent local recurrences should not be a contraindication to breast reconstruction in selected patients. PMID- 6280665 TI - Protozoan and viral infections of feral cats. AB - Identification of protozoan oocysts and serological tests were used to determine the prevalence of infections among 300 mainly adult feral cats in three different habitat types in south-eastern Australia. Oocysts of Isospora rivolta and Isospora felis were recovered from 3% and 4% respectively of 300 feral cat samples. Haemagglutination inhibition antibody to Toxoplasma gondii was detected in 20% of 75 cat sera tested. A high prevalence of specific antibody to feline panleukopaenia virus (79%) and feline calici virus (77%) was demonstrated but the prevalence of antibody to feline herpes virus was low (11%). 15 strains of feline calici virus were isolated from pharyngeal swabs. There were no other virus isolations from the 60 pharyngeal and rectal swabs taken. These viral and protozoan infections could not be linked with any obvious pathological conditions. Most cats were in good condition with light to moderate fat stores in depot areas. Limb fractures and other skeletal abnormalities occurred infrequently. Major tooth damage or absence of important teeth was evident in about 20% of 164 animals examined. There was no correlation between major tooth damage and poor body condition. PMID- 6280666 TI - Relaxin--a review. AB - Relaxin is a polypeptide hormone, similar in structure to insulin and has been found in the female of all species studied. The corpus luteum of pregnancy is the main source of relaxin in many species but in others the decidua is apparently of greater importance. It has also been found in other tissues; e.g. prostatic fluid, testis and ovary. First discovered and extracted from the corpora lutea of pregnant sows in an impure form in 1926, it was found to relax the pubic ligament of the oestrogen primed guinea-pig. It was named after this action, but has since been found to have many other possible roles, including preparation of the endometrium for implantation, inhibition of uterine activity in early pregnancy, remodelling of the uterine stroma during pregnancy, cervical ripening and the initiation of parturition. Relaxin's main cellular action in pregnancy may be to drive collagen biosynthesis in its target organs, thus facilitating the remodelling of the connective tissue. Due to the impurity of relaxin preparations used in clinical trials until the mid-1970's, the role of relaxin in the human has been in doubt. Porcine and rat relaxins have now been highly purified and their detailed structure is known. Human relaxin awaits adequate isolation, purification and characterization, and is not yet available for laboratory and clinical trials. However, the recent preparation of purified porcine relaxin for clinical trials and the availability of specific radioimmunoassays for this relaxin together with the identification of relaxin receptor sites, are rapidly helping to establish the concept that relaxin is indeed an important hormone both in human reproduction and in other physiological processes. PMID- 6280667 TI - Orally administered THIP inhibits food intake in the rat. PMID- 6280668 TI - [Synovectomy in rare forms of arthritis]. PMID- 6280669 TI - Characterization of a rat liver cyclic GMP-activated phosphodiesterase by chromatography on hexyl-agarose. Inhibition of phosphodiesterase activity by hexyl-agarose. AB - Chromatography on hexyl-agarose resolved a partially purified cyclic GMP activated phosphodiesterase from rat liver into two peaks of activity: the first was eluted with 0.5 M-KCl and was cyclic AMP-specific. The second was tightly bound to hexyl-agarose and was not eluted with KCl (0--2.0 M), which enhanced the hydrophobic interactions of this form with the matrix. It was eluted with 0.5 M Tris, hydrolysed cyclic AMP and cyclic GMP and was specifically activated by cyclic GMP. The cyclic GMP-activated phosphodiesterase was immobilized on hexyl agarose. Enzyme activity, quantitatively bound to hexyl-agarose, was not released from the hydrophobic matrix in the presence of cyclic AMP or cyclic GMP, under our assay conditions. The immobilized form of the enzyme retained catalytic activity, was inhibited by 0.1 mM-cyclic AMP and was activated by micromolar concentrations of cyclic GMP to a lesser extent (7-fold) than the control, i.e. the enzyme mixed with unsubstituted agarose (15-fold). When the enzyme was immobilized, inhibition of cyclic AMP phosphodiesterase activity was only observed in the presence of cyclic GMP (at 3 microM); in its absence, activity remained unchanged. The kinetic behaviour of the immobilized enzyme is consistent with the hypothesis of a binding site distinct from the hydrolytic and activating sites. PMID- 6280670 TI - The stereochemical course of hydrolysis catalysed by snake venom 5'-nucleotide phosphodiesterase. AB - Adenosine 5'-(S)-[16O,17O,18O]phosphate was pyrophosphorylated by the combined action of adenylate kinase and pyruvate kinase. The isotopomers of adenosine 5' [alpha-16O,17O,18O]triphosphate were hydrolysed by venom 5'-nucleotide phosphodiesterase (Crotalus adamanteus) in H2(17)O. Analysis by 31P nuclear magnetic resonance spectroscopy of the resulting adenosine 5' [16O,17O,18O]phosphate, after cyclization and esterification, showed that the hydrolysis occurs with retention of configuration at phosphorus. The most likely explanation of this observation is that the enzymic hydrolysis involves a double displacement at phosphorus with a covalent nucleotidyl--enzyme intermediate on the reaction pathway. PMID- 6280671 TI - Design, synthesis and DNA-binding capacity of a new peptidic bifunctional intercalating agent. AB - A lysyl-lysine bifunctional derivative of 9-aminoacridine has been synthesized and its DNA-binding capacity established by electron-paramagnetic-resonance study. For this purpose the binding parameters of a spin-labelled aminoacridine probe were estimated and the affinities of the lysylacridinyl-lysyldiamino-octane dimer and of 9-amino-acridine could be evaluated by competitive assays. The competition study provided quantitative results concerning the dissociation constant (KD) of the dimer. The obtained value was closely similar to the KD of 9 aminoacridine determined by the same method and to the KD previously reported for the anti-tumour and antibiotic bifunctional intercalator quinomycins. PMID- 6280673 TI - Purification and properties of bovine caeruloplasmin. AB - A novel method is reported for isolation of bovine caeruloplasmin from plasma; it involves a rapid and mild procedure, namely two column chromatographies with stepwise elution and one (NH4)2SO4 precipitation, and results in a proteolytically undegraded homogeneous protein. The general structure of the protein, as evaluated by molecular-weight determination and amino acid composition, is very similar to that established for human and rat caeruloplasmin. Copper determination and e.p.r. spectral analysis on the native and NO-treated protein gave a metal-to-protein stoichiometry of six atoms of copper per molecule. Three copper atoms were detectable by e.p.r., with Type 2/Type 1 ratio = 1 : 3 in most samples. The protein is very sensitive to storage and/or handling. A component was isolated from aged samples, which was found to contain approximately four copper atoms per 125000 daltons, two of which were detectable by e.p.r. with the characters of Type 2 copper. However, the same component was found to be present, although to a lesser extent, in the fresh preparation and does not seem to be related to proteolytic degradation. This component has no oxidase activity. On the basis of these results it is suggested that caeruloplasmin molecules are intrinsically heterogeneous with respect to both copper content and copper type, and this can explain the intriguing stoichiometry regarding the different types of copper centres. PMID- 6280672 TI - Coupling of [33S]sulphur to molybdenum(V) in different reduced forms of xanthine oxidase. AB - Different reduced forms of xanthine oxidase, labelled specifically in the cyanide labile site with 33S, were prepared and examined by electron paramagnetic resonance. Coupling of this isotope to molybdenum(V) was quantified with the help of computer simulations and found to differ markedly from one reduced form to another. The xanthine Very Rapid signal shows strong, highly anisotropic, coupling with A(33S)av. 1.27 mT. For this signal, axes of the g- and A(33S) tensors are rotated relative to one another. One axis of the A-tensor is in the plane of gxx ang gyy, but rotated by 40 degrees relative to the gxx axis, whereas the direction of weakest coupling to sulphur deviates by 10 degrees from the gzz axis. In contrast with this signal, only rather weaker coupling was observed in different types of Rapid signal [A(33S)av. 0.3--0.4 mT], and in the Inhibited signal coupling was weaker still [A(33S)av. 0.1--0.2 mT]. Clearly, there must be substantial differences in the structures of the molybdenum centre in the different signal-giving species, with the sulphur atom perhaps in an equatorial type of ligand position in the Very Rapid species but in a more axial one in the other species. Structures are discussed in relation to the mechanism of action of the enzyme and the nature of the proton-accepting group that participates in turnover. PMID- 6280674 TI - Preparation of selectively metal-free and metal-substituted derivatives by reaction of Cu--Zn superoxide dismutase with diethyldithiocarbamate. AB - Incubation of Cu--Zn superoxide dismutase with diethyldithiocarbamate at increasing ligand/protein ratios and subsequent high-speed centrifugation led to proportional removal of copper from the protein, at variance with previous results [Misra (1979) J. Biol. Chem. 254, 11623--11628]. No zinc was lost, even at very high excesses of chelating agent. In this way a copper-free protein could be readily prepared, with avoidance of the critical pH condition and the dialysis step required in a previous method employing cyanide. The holoprotein was fully reconstituted from the copper-free protein by stoicheiometric re-addition of copper. From the mixture of metal-depleted forms originated by treatment with slight diethyldithiocarbamate excess, the protein containing copper only on one subunit, [Cu1--Zn2], could be isolated by preparative column electrophoresis. This species reproducibly showed 25% more specific activity (catalytic constant per copper) than that of the native or reconstituted [Cu2--Zn2] protein. This may result from long-range conformational effects between the active sites. By adding Co2+ ions to the vacant copper site of [Cu1--Zn2] a hybrid molecule containing Cu(II) on one subunit and Co(II) in the homologous site of the other subunit was prepared. Its activity, referred to copper, was identical with that of the native protein. PMID- 6280675 TI - Kinetic solvent isotope effects on the deacylation of specific acyl-papains. Proton inventory studies on the papain-catalysed hydrolyses of specific ester substrates: analysis of possible transition state structures. AB - 1. The hydrolyses of the p-nitrophenyl esters of N-benzyloxycarbonylglycine, alpha-N-benzyloxycarbonyl-L-lysine and N-methoxycarbonyl-L-phenylalanylglycine catalysed by papain (EC 3.4.22.2) have been studied in solvents having a variable composition of 2H2O and H2O. 2. kcat., which represents deacylation in the papain catalysed hydrolysis of reactive esters, is some 2.3-fold less in 2H2O compared with H2O. The magnitude of kcat. has been determined as a function of the 2H atom fraction of the solvent. 3. Both linear and non-linear methods of least-square regression analysis have been applied to the data in order to obtain best-fit parameter values for several three-parameter models which express kcat. in terms of the 2H atom fraction of the solvent. These models represent some possible modes of restructuring of the active site protonic configuration consequent upon transition state formation. 4. The results of curve fitting reveal an essentially linear dependence of kcat. upon the 2H atom fraction, and it may therefore be concluded that the isotope effect originates from a single proton which is in the process of transfer in the transition state. 5. It is postulated on the basis of this and other evidence that the mobile proton is transferred from an attacking water molecule to the imidazole side chain of His-159 during tetrahedral intermediate formation. This has the effect of stabilizing the transition state and promoting catalysis. The role of His-159 in deacylation is therefore to provide general base catalysis. 6. Models that involve two or more protons, such as a two-proton relay system analogous to that proposed for the serine proteinases, or a multiproton 'medium' effect, are considered unlikely on the basis of the data reported in this paper. 7. A more detailed examination of possible transition state structures reveals that the only structure compatible with available experimental data and consistent with certain theoretical predictions is one in which the proton translocated in concern with reorganization of the heavy atom framework. In addition, the transition state vibrations of the mobile proton are strongly coupled to those of the heavy atoms. These properties of the transition state are also manifest in the transition state for the deacylation of serine proteinases. PMID- 6280676 TI - The EcoRI restriction endonuclease, covalently closed DNA and ethidium bromide. AB - The reactions of the EcoRI restriction endonuclease on the covalently closed DNA of plasmid pMB9 were studied in the presence of ethidium bromide. At the concentrations of ethidium bromide tested, which covered the range over which the DNA is changed from negatively to positively supercoiled, the dye caused no alteration to the rate at which this enzyme cleaved the covalently closed DNA to yield the open-circle form, but the rate at which these open circles were cleaved to the linear product could be inhibited. The fluorescence change, caused by ethidium bromide binding with different stoichiometries to covalently closed and open-circle DNA, provided a direct and sensitive signal for monitoring the cleavage of DNA by this enzyme. This method was used for a steady-state kinetic analysis of the reaction catalysed by the EcoRI restriction enzyme. Reaction mechanisms where a complex between DNA and Mg2+ is the substrate for this enzyme were eliminated, and instead DNA and Mg2+ must bind to the enzyme in separate stages. The requisite controls for this fluorimetric assay in both steady-state and transient kinetics studies, and its application to other enzymes that alter the structure of covalently closed DNA, are described. PMID- 6280677 TI - NAD+, ADP-ribosylation and transcription in permeabilized mammalian cells. AB - When permeabilized hamster fibroblasts were incubated with 4 mM-NAD+, the substrate for poly(ADP-ribose) polymerase, RNA polymerase I activity was inhibited by about 85%. This inhibition was not relieved by prior incubation of cells with 3-aminobenzamide, a potent inhibitor of the poly(ADP-ribose) polymerase. Digestion of cells with pancreatic deoxyribonuclease I resulted in the inhibition of RNA polymerase I by 80% and the activation of poly(ADP-ribose) polymerase by up to 300%; prior incubation with 3-aminobenzamide did not prevent the inhibition of the RNA polymerase activity. No radioactivity was found associated with RNA polymerase I during later stages of purification of this enzyme from permeabilized cells previously incubated with [14C]NAD+. The inhibitory effect of NAD+ on RNA polymerase I was not specific for NAD+, as other small, negatively charged molecules with a nuclear location also inhibited the enzyme. The results do not support the concept of a role for ADP-ribosylation in transcription catalysed by RNA polymerase I. PMID- 6280678 TI - Sequencing long DNA fragments cloned in bacteriophage M13 by using internal primers. The sequence analysis of a yeast DNA fragment containing a replication origin. AB - In the ;shotgun' procedure for sequencing DNA, DNA fragments are cloned into a phage M13 vector and sequenced by using a flanking primer. In a variation of this procedure a longer DNA sequence is cloned into M13, the two single-stranded recombinants identified and sequenced by using a set of internal primers prepared by exonuclease III digestion of restriction fragments. PMID- 6280679 TI - The hypothesis of localized chemiosmosis is unsatisfactory. AB - The hypothesis of 'localized' chemiosmosis have been put to an experimental test in the system of oxidative phosphorylation by rat liver mitochondria. We find that the variation of the ratio of phosphate potential to delta muH with delta muH does not depend on how delta muH is varied. This is in conflict with hypotheses of localized chemiosmosis. Of all coupling hypotheses, only the parallel-coupling hypothesis can explain the observations, unless variation of the H+/ATP stoichiometry of the ATPase proton pump is accepted. In the latter case. 'macroscopic' chemiosmosis can explain the observations equally well as hypotheses of localized chemiosmosis. It is concluded that either variation of H+/ATP stoichiometries must be accepted, or that the parallel-coupling hypothesis should be reformulated so that it becomes open to experimental tests. PMID- 6280680 TI - Solubilization of the ileal receptor for intrinsic factor--vitamin B-12 complex by digestion with papain. AB - Brush-border-membrane vesicles isolated from hamster ileum were incubated with either papain or Pronase P and subsequently centrifuged to obtain soluble (supernatant) and insoluble (pellet) fractions. Papain (4 units/ml) solubilized 95--100% of the sucrase and leucine naphthylamide-hydrolysing activities but only 30% of the alkaline phosphatase. Digestion with papain also resulted in the solubilization of more than 75% of the ileal receptor for intrinsic factor vitamin B-12 complex with a corresponding decrease in receptor activity in the pellet. Essentially 100% of the receptor activity was recovered. In contrast, digestion with Pronase P resulted in a decrease in total receptor activity. Papain-solubilized receptor was not sedimented by centrifugation at 105 000 g for 90 min and was eluted in the included volume of Sepharose 6B. Like the binding to more intact preparations, binding of intrinsic factor-vitamin B-12 complex to papain-solubilized receptor was rapid, reaching 50% of maximum in 8 min, and required Ca2+. Although Mg2+ could not completely substitute for Ca2+, Mg2+ did stimulate Ca2+-dependent binding at low Ca2+ concentrations. These results demonstrate that the ileal receptor for intrinsic factor-vitamin B-12 complex can be solubilized with papain, and suggest that papain solubilization may be a useful first step in the isolation and purification of this receptor. PMID- 6280681 TI - Effects of chronic modification of dietary fat and carbohydrate in rats. AB - 1. Rats were fed on diets enriched with starch, sucrose, corn oil or beef tallow for 3 weeks and the activities of various enzymes in the liver were measured. 2. The mitochondrial glycerol phosphate acyltransferase activity was lower in rats fed on the starch diet than on the two high-fat diets. 3. The non-microsomal (presumably peroxisomal) dihydroxyacetone phosphate acyltransferase activity was higher in rats fed on the starch diet and corn-oil diets than in those fed on the sucrose and beef-tallow diets. Urate oxidase activity was higher in rats fed on the starch diet than in the three other groups. There were no significant differences in the activity of acyl-CoA oxidase among the groups. 4. The activity of soluble phosphatidate phosphohydrolase was not significantly different among the dietary groups. There were increases of 3.3--4.3-fold in this activity in the dietary groups 6h after injection of corticotropin. The equivalent increases for the mitochondrial glycerol phosphate acyltransferase were 1.4--1.6 fold. 5. The corticosterone responses to the corticotropin injection were not significantly different between dietary groups. However, the corticosterone response of the rats fed on the two high-fat diets was prolonged when the rats were given an acute load of fructose [Brindley, Cooling, Glenny, Burditt & McKechnie (1981) Biochem. J. 200. 275--283]. 6. Rats fed on the high-fat diets had higher concentrations of circulating cholesterol than those fed on the starch and sucrose diets. Serum triacylglycerol concentrations were lower in the rats fed on the starch diet than in the three other groups. 7. The results are discussed in terms of the relationship between diet, hormonal balance and hepatic glycerolipid metabolism. PMID- 6280682 TI - The activities of lipoprotein lipase and of enzymes involved in triacylglycerol synthesis in rat adipose tissue. Effects of starvation, dietary modification and of corticotropin injection. AB - 1. The effects of dietary modification, including starvation, and of corticotropin injection on the activities of acyl-CoA synthetase, glycerol phosphate acyltransferase, dihydroxyacetone phosphate acyltransferase, phosphatidate phosphohydrolase, diacylglycerol acyltransferase and lipoprotein lipase were measured in adipose tissue. 2. Lipoprotein lipase activities in heart were increased and those in adipose tissue were decreased when rats were fed on diets enriched with corn oil or beef tallow rather than with sucrose or starch. The lipoprotein lipase activity was lower in the adipose tissue of rats fed on the sucrose rather than on the starch diet. 3. Rats fed on the beef tallow diet had slightly higher activities of the total glycerol phosphate acyltransferase in adipose tissue than did rats fed on the sucrose or starch diet. The diacylglycerol acyltransferase and the mitochondrial glycerol phosphate acyltransferase activities were higher for the rats fed on the tallow diet than for those fed on the corn-oil diet. 4. Starvation significantly decreased the activities of lipoprotein lipase (after 24 and 48 h), acyl-CoA synthetase (after 24 h) and of the mitochondrial glycerol phosphate acyltransferase and the N ethylmaleimide-insensitive dihydroxyacetone phosphate acyltransferase (after 48 h) in adipose tissue. The activities of the microsomal glycerol phosphate acyltransferase, diacylglycerol acyltransferase and the soluble phosphatidate phosphohydrolase were not significantly changed after 24 or 48 h of starvation. 5. The activities of lipoprotein lipase and phosphatidate phosphohydrolase in adipose tissue were decreased 15 min after corticotropin was injected into rats during November to December. No statistically significant differences were found when these experiments were performed during March to September. These differences may be related to the seasonal variation in acute lipolytic responses. 6. These results are discussed in relation to the control of triacylglycerol synthesis and lipoprotein metabolism. PMID- 6280683 TI - Binding of nitrobenzylthioinosine to high-affinity sites on the nucleoside transport mechanism of HeLa cells. AB - Nitrobenzylthioinosine (NBMPR) binds reversibly, but with high affinity (Kd 0.1- 1.2 nM), to inhibitory sites on nucleoside-transport elements of the plasma membrane in a variety of animal cells. The present study explored relationships in HeLa cells between NBMPR binding and inhibition of uridine transport. The Km value for inward transport of uridine by HeLa cells in both suspension and monolayer culture was about 0.1 mM. The affinity of the transport-inhibitory sites for uridine (Kd 1.7 mM), inosine (Kd 0.4 mM) and other nucleoside permeants was low relative to that for NBMPR. The pyrimidine homologue of NBMPR, nitrobenzylthiouridine, also exhibited low affinity for the NBMPR-binding sites. Pretreatment of HeLa cells with p-chloromercuribenzene sulphonate (p-CMBS) or N ethylmaleimide (NEM) decreased binding of NBMPR to its high-affinity sites and inhibited uridine transport, indicating the presence of thiol groups essential to both processes. NEM, a more penetrable reagent than p-CMBS, inhibited binding and transport at much lower concentrations than the latter compound. Pretreatment of cells with concentrations of p-CMBS that alone had no effect on either NBMPR binding or uridine transport increased the sensitivity of transport to NBMPR inhibition and changed the shape of the NBMPR concentration-effect curve, suggesting synergistic inhibiton of uridine-transport activity by these two agents. PMID- 6280684 TI - Light activation of fructose bisphosphatase in photosynthetically competent pea chloroplasts. AB - The fructose bisphosphatase (EC 3.1.3.11) activity of type A chloroplasts isolated from young (9-day-old) pea (Pisum sativum var. Progress no. 9) plants, assayed at physiological pH, substrate and Mg2+ concentrations, increased rapidly on illumination. The enzyme activity detected was more than sufficient to account for observed rates of Co2 fixation both during the induction period and during steady-state CO2 fixation, whether or not dihydroxyacetone phosphate had been added to the preparation. Omission of catalase from the suspension medium had no effect. On switching off the light, CO2 fixation by the chloroplasts ceased at once, yet fructose bisphosphatase activity decreased much more slowly. Changes in enzyme activity were much less marked if assays were conducted at 3 mM substrate and 10 mM-Mg2+. Chloroplasts from older (13--20-day-old) peas only fixed CO2 rapidly if catalase was present in the assay medium. The fructose bisphosphatase activity detected under physiological assay conditions was again more than sufficient to account for observed rates of Co2 fixation. In the presence of added dihydroxyacetone phosphate, however, the rate of Co2 fixation appeared to be determined by the rate of light activation of fructose bisphosphatase. In general, the rates of Co2 fixation and enzyme activation, and the final enzyme activity achieved, decreased markedly with increasing age of the plants. The role of light activation of fructose bisphosphatase as a means of controlling the rate of CO2 fixation in pea chloroplasts is discussed. PMID- 6280685 TI - Adenosine production inside rat polymorphonuclear leucocytes. AB - Adenosine synthesis was studied during 2-deoxyglucose-induced ATP catabolism in intact rat polymorphonuclear leucocytes. When both adenosine kinase (EC 2.7.1.20) and adenosine deaminase (EC 3.5.4.4) were selectively inhibited, adenosine accumulated. Adenosine formation took place inside the intact cells by a metabolic pathway independent of the ecto-5'-nucleotidase (EC 3.1.3.5). Distinct metabolic pathways are proposed for adenosine production from intracellular or extracellular nucleotides. PMID- 6280686 TI - Inhibitory action of adenosine 3',5'-monophosphate on phosphatidylinositol turnover: difference in tissue response. PMID- 6280687 TI - On the presence of adenosine 3',5'-cyclic monophosphate in moss (Funaria hygrometrica). PMID- 6280688 TI - Isolated Entamoeba histolytica actin does not inhibit DNAse-I activity. PMID- 6280689 TI - Increased filamentous actin in islets of Langerhans from fasted hamsters. PMID- 6280691 TI - Adriamycin inactivates cytochrome c oxidase by exclusion of the enzyme from its cardiolipin essential environment. PMID- 6280690 TI - Dissociation by piroxicam of degranulation and superoxide anion generation from decrements in chlortetracycline fluorescence of activated human neutrophils. PMID- 6280692 TI - The tissue distribution of fructose-2,6-p2 and fructose-6-P,2-kinase in rats and the effect of starvation diabetes and hypoglycemia on hepatic fructose-2,6-P2 and fructose-6-P,2-kinase. PMID- 6280693 TI - Structural alteration of rRNA in the L7/L12 region of 50s ribosome on removal of L7/L12 proteins. PMID- 6280694 TI - The Na+-K+-pump, energy metabolism, and obesity. PMID- 6280695 TI - Release and activation of phosphorylase phosphatase upon rupture of organelles from rat liver. PMID- 6280696 TI - Inhibition of topoisomerase I by heparin. PMID- 6280697 TI - Dehydro-enkephalins. IV. Discriminative recognition of delta and mu opiate receptors by enkephalin analogs. PMID- 6280698 TI - Isolation of QP-C and reconstitution of the QH2-c reductase. PMID- 6280699 TI - Existence of lysine-derived cross-linking in connectin, an elastic protein in muscle. PMID- 6280700 TI - Purification and characterization of calmodulin from rat liver mitochondria. PMID- 6280701 TI - Cerebral cortical microvessels: an insulin-sensitive tissue. PMID- 6280702 TI - Human histone genes are interspersed with members of the Alu family and with other transcribed sequences. PMID- 6280703 TI - Characterisation of human glomerular basement membrane antigenic fractions isolated by affinity chromatography utilising anti-glomerular basement membrane autoantibodies. PMID- 6280704 TI - Inhibition of mouse mastocytoma protein kinases by amiloride. PMID- 6280705 TI - Reciprocal changes in fructose-6-phosphate,2-kinase and fructose-2,6 bisphosphatase activity in response to glucagon and epinephrine. PMID- 6280706 TI - Prostaglandin A1 and E1 inhibit the plating efficiency and proliferation of murine melanoma cells (Cloudman S-91) in soft agar. PMID- 6280707 TI - Binding of benzo(a)pyrene to hepatic cytosolic protein enhances its microsomal oxidation. PMID- 6280708 TI - Ionic strength and pH effects in the kinetically controlled synthesis of benzylpenicillin by nucleophilic deacylation of free and immobilized phenyl acetyl-penicillin amidase with 6-aminopenicillanic acid. PMID- 6280709 TI - Activation/inactivation of human angiotensin I converting enzyme following chemical modifications of amino groups near the active site. PMID- 6280710 TI - Effects of estrogens on aryl hydrocarbon hydroxylase mediated binding of benzo(a) pyrene metabolites to DNA in vitro. PMID- 6280711 TI - Enzymatic formation of 14,15-leukotriene A and C(14)-sulfur-linked peptides. PMID- 6280712 TI - ATP(adenosine triphosphate)-vanadyl complex. PMID- 6280714 TI - The effect of the Q base modification on the usage of tRNAHis in globin synthesis. PMID- 6280713 TI - Na+, K+-ATPase: evidence for the binding of ATP to the phosphoenzyme. PMID- 6280715 TI - Inhibition of canine lung angiotensin converting enzyme by ACTH and structurally related peptides. PMID- 6280716 TI - DNA methylation levels in normal and chemically-transformed mouse 3T3 cells. PMID- 6280717 TI - Inhibition of leukotriene B4 synthesis in human polymorphonuclear leukocytes after exposure to meningococcal lipopolysaccharide. PMID- 6280718 TI - Glycosylation of thyroglobulin and response to thyrotropin of human cultured thyroid cells. PMID- 6280719 TI - Azidoacridines as photoaffinity probes for ionic channels in excitable membranes. PMID- 6280720 TI - Endogenous protein phosphorylation in adhesive plaques of substrate-attached fibroblasts. PMID- 6280721 TI - Cis- and trans-diamminedichloroplatinum(II) binding products different tertiary structural changes on SV40 DNA. PMID- 6280722 TI - Immunosuppressive properties of leukotriene D4 and E4 in vitro. PMID- 6280723 TI - Calcium uptake by enzyme-treated permeabilized myometrial cells. PMID- 6280724 TI - Iron-EDTA stimulated reduction of indicine N-oxide by the hepatic microsomal fraction, isolated hepatocytes, and the intact rat. AB - Fe(III) complexes of EDTA and diethylenetriamine pentaacetic acid (DETAPAC) at low concentrations (between 1 and 100 microM) produced up to a 20-fold increase in anaerobic microsomal NADPH- and NADH-dependent reduction of indicine N-oxide. Under aerobic conditions microsomal indicine N-oxide reduction was stimulated to half the levels seen under anaerobic conditions. EDTA alone was much less effective at stimulating indicine N-oxide reduction, while FeCl3 alone had no effect on reduction. Other complexes of Fe(III) had little or no effect in stimulating microsomal indicine N-oxide reduction. Fe(III)-EDTA stimulated indicine N-oxide reduction by purified NADPH-cytochrome P-450 reductase and NADPH. It is probable that iron serves to transfer electrons between microsomal flavoprotein reductases and indicine N-oxide. The redox potential and the presence of an exchangeable ligand, such as water, in the inner ligand sphere of the iron complex are suggested to be important factors in determining which iron complexes will stimulate indicine N-oxide reduction. EDTA complexes of other transition metal ions do not stimulate indicine N-oxide reduction. Hydroxyl radicals, detected as the spin adduct of 5,5-dimethyl-1-pyroline-N-oxide, appear to be formed during Fe(II)-EDTA-dependent reduction of indicine N-oxide under anaerobic conditions. Fe(III)-EDTA at concentrations between 50 and 250 microM stimulated indicine N-oxide reduction by rat isolated hepatocytes up to 5-fold under anaerobic conditions and to half these values under aerobic conditions. By themselves, EDTA and FeCl3 at similar concentrations produced a small stimulation of indicine N-oxide reduction by hepatocytes under anaerobic conditions. Fe(III) EDTA stimulated indicine N-oxide reduction by murine leukemia P-388 cells under aerobic conditions and by rat caecal flora under anaerobic but not aerobic conditions. Fe(III)-EDTA, EDTA or FeCl3 administered to rats produced a 3-fold increase in the 24-hr urinary excretion of indicine following an i.p. dose of indicine N-oxide. PMID- 6280725 TI - Role of catalase and hydroxyl radicals in the oxidation of methanol by rat liver microsomes. AB - In view of the presence of adventitious catalase in isolated microsomes, and the requirement for H2O2, it has been suggested that NADPH-dependent oxidation of methanol by rat liver microsomes was mediated exclusively by the peroxidatic activity of catalase. However, H2O2 may also serve as a precursor of the hydroxyl radical, and methanol reacts with hydroxyl radicals to produce formaldehyde. Inhibition of H2O2 production should therefore decrease methanol oxidation by either a hydroxyl radical-dependent pathway or a catalase-dependent pathway. To attempt to clarify some of the controversies concerning the roles of H2O2 and catalase in the microsomal pathway of oxidation of short chain alcohols, studies were carried out to determine the nature of the pathway responsible for methanol oxidation by isolated microsomes. In the absence of the catalase inhibitor azide, methanol may be oxidized primarily by the peroxidatic activity of catalase since there was little effect on methanol oxidation by competing hydroxyl radical scavengers. Azide, which inhibited catalase activity greater than 95%, inhibited NADPH-dependent oxidation of methanol by 30-50%. The azide-insensitive (catalase independent) pathway of methanol oxidation was inhibited by scavengers of hydroxyl radicals. The inhibition of the scavengers reflected the rate constant for interaction with hydroxyl radicals and was greater at lower concentrations of methanol than at higher concentrations, suggesting competition between the scavengers and methanol. The addition of H2O2 stimulated the oxidation of methanol in the presence of azide; H2O2 may serve as a precursor of the hydroxyl radical. Iron-EDTA, which is known to increase hydroxyl radical production, stimulated the oxidation of methanol in the absence and presence of azide. The stimulation by iron-EDTA was blocked by the competing hydroxyl radical scavengers even in the absence of azide, suggesting that the added iron-EDTA favorably with microsomal catalase for H2O2 to produce hydroxyl radicals (or a species with the oxidizing power of the hydroxyl radical). These results suggest that in microsomes, depending on the absence or presence of azide, methanol may be oxidized by two primary pathways, one involving the peroxidatic activity of catalase, and the other in which hydroxyl radicals, generated from microsomal electron transfer, play a role. In view of the crucial role played by H2O2 in both pathways, inhibition of H2O2 formation should not be interpreted solely as evidence for a role for catalase in the microsomal oxidation of alcohols. PMID- 6280726 TI - Early decrease in secretin-, glucagon-, and isoproterenol-stimulated cardiac adenylate cyclase activity in rats treated with isoproterenol. AB - Male Wistar albino rats received three times daily for one to five days 0.25 to 5.0 mg/kg D,L-isoproterenol intraperitoneally. D.L-Isoproterenol injections provoked a time dependent- and dose-related cardiac hypertrophy. With moderate hypertrophy, a selective decrease in secretin-stimulated adenylate cyclase activity occurred. When heart hypertrophy was more pronounced, greater losses in secretin-, as well as in D,L-isoproterenol-, glucagon-, guanine nucleotide-, and fluoride-stimulated enzyme activity developed. Hormone stimulations of adenylate cyclase were more severely, curtailed (60 to 65%) than guanine nucleotide or fluoride stimulations (40 to 45%). The accompanying loss in beta-receptors (35%) was proportionately lower than the loss of D,L-isoproterenol sensitivity of adenylate cyclase. This complex pattern of adenylate cyclase desensitization in heart membranes from animals chronically treated with D,L-isoproterenol is reminiscent of that observed in heart membranes from spontaneously hypertensive rats. PMID- 6280727 TI - Trichlorethylene and halothane inhibit uptake of 5-hydroxytryptamine in the isolated perfused rat lung. AB - Lung uptake of 5-hydroxytryptamine (5-HT) was determined in isolated perfused and ventilated rat lung, and was found to decrease with time according to a two compartmental model. When the lungs were exposed to either trichlorethylene (TRI) or halothane, the uptake of 5-HT was drastically reduced. Both TRI and halothane gave log dose inhibition curves, which were superimposed, i.e. they were equally potent to inhibit lung uptake of 5-HT. At a concentration TRI of 18,000 ppm, the extraction of 5-HT was inhibited by 80 +/- 2 (X +/- S.E.M.) per cent, at 8500 ppm the inhibition was 65 +/- 6 per cent and 25 +/- 1 per cent at 3000 ppm. When the lungs were exposed to halothane, the inhibition was 85 +/- 6 per cent at 40,000 ppm, 48 +/- 1 per cent at 6000 ppm, and 15 +/- 0.3 per cent at 2000 ppm. When exposure to the solvent was discontinued, extraction of 5-HT was rapidly normalized. There was no detectable displacement of [3H]-5-HT from lungs saturated with the amine when they subsequently were exposed to solvent containing atmosphere. This inhibition of lung uptake of 5-HT from the circulation is therefore postulated as to be an effect dependent on concentration solvent in the tissue, and is probably due to a reversible membrane stabilization of the endothelium. PMID- 6280728 TI - Histamine H2 receptors in rat renal glomeruli. AB - The aim of this study was to demonstrate histamine-H2 receptors in glomeruli isolated from rat renal cortex and to correlate binding to stimulation by histamine of glomerular cyclic AMP concentration. Binding studies were performed at 10-12 degrees C using [3H]cimetidine as a tracer. Specificity of binding relies on the following: inhibition of [3H]cimetidine binding by the unlabelled drug, other H2-antagonists and agonists in contrast with the very weak inhibitory effects of H1 agonists and antagonists; reversibility of steady-state binding after addition of unlabelled drug; half inhibition of the glomerular cyclic AMP response to histamine at concentrations of cimetidine close to the KD value derived from the binding studies (3 microM); calculated KD value in agreement with the therapeutical concentration of cimetidine and the physiological concentration of histamine. [3H]Cimetidine binding concentration of cimetidine and the physiological concentration of histamine. [3H]Cimetidine binding strikingly increased in the presence of copper chloride (20-300 microM) due to an increase both in number of sites and affinity. However this greater binding did not influence either the inhibitory effect of cimetidine on histamine-induced glomerular cyclic AMP concentration or the stimulatory effect of histamine itself. [3H]Cimetidine binding was temperature-dependent since it progressively diminished from 0 to 37 degrees. This was not due to [3H]cimetidine degradation as shown by thin layer chromatography but rather to a change in drug-receptor interaction at higher temperatures. Glumerular concentration of cyclic AMP increased progressively in the presence of histamine (0.1-1000 microM). This stimulatory effect was markedly inhibited by H2 antagonists. These data demonstrate the presence in rat glomeruli of H2 receptors linked to adenylate cyclase. PMID- 6280729 TI - Enzymatic changes in the male reproductive organs by delta-9 tetrahydrocannabinol. AB - Like most psychoactive agents, cannabis and its active component delta-9 tetrahydrocannabinol (delta 9-THC) have been reported to affect the neuroendocrine axis in animals. The effect of delta 9-THC on some of the functionally important enzymes of the male reproductive organs are reported. The study indicates that delta 9-THC reduces the activities of the enzymes, beta glucuronidase, alpha-glucosidase acid phosphatase and fructose-6-phosphatase in a dose related manner in the testis, prostate as well as in the epididymis. It may be concluded that delta 9-THC may interfere with the normal functioning of the male reproductive organs. PMID- 6280730 TI - Inhibition of calmodulin-activated cyclic nucleotide phosphodiesterase: multiple binding-sites for tricyclic drugs on calmodulin. AB - A cyclic nucleotide phosphodiesterase from guinea-pig heart is activated by calmodulin in the presence of calcium ions. Activation was measured over a range of calmodulin concentrations, and is antagonised by several tricyclic psychotropic drugs including trifluoperazine, imipramine, chlorpromazine and amitriptyline. When the concentration of amitriptyline was increased, its apparent inhibition constant for binding to calmodulin decreased. This was due in part to binding of amitriptyline to glass surfaces; but after correction for this the discrepancy was still significant. It is proposed that this is due to two sites on calmodulin for amitriptyline, with binding to either site being sufficient to prevent calmodulin from activating phosphodiesterase. PMID- 6280731 TI - Inhibition of some polymorphonuclear leukocyte functions by ethacrynic acid. AB - Ethacrynic acid (10(-4) M) inhibits exocytosis, phagocytosis and superoxide release in rabbit polymorphonuclear leukocytes (PMN's). Dihydroethacrynic acid is a much weaker inhibitor of these PMN functions. Though ethacrynic acid inhibits ATPase activity in the PMN, this occur at much higher concentrations than required for inhibition of exocytosis and superoxide release, thus a causal relationship seems unlikely. The same applies to inhibition of ATP generation by ethacrynic acid: the concentration required to decrease ATP level in PMN's is much higher than required for the inhibitory effect on exocytosis. Inhibition of exocytosis by ethacrynic acid can be prevented by dithiothreitol. It is concluded that vulnerable sulfhydryl groups are involved in the inhibition by ethacrynic acid. PMID- 6280732 TI - Effect of lithium on synaptosomal brain enzymes. AB - During 7 days of lithium administration to rats, the levels of adenylate cyclase bound to brain synaptosomes decreased, while those of acetylcholinesterase increased. Both enzyme levels returned to their initial values after 1 month of treatment. Monoamine oxidase, which is bound to a mitochondrial membrane, was not affected by lithium treatment. Arrhenius plots of the brain synaptosomal enzymes (Na + K)ATPase and acetylcholinesterase from rats treated with LiCl for 7 or 30 days showed a lower transition temperature. Also, when these synaptosomal enzymes were exposed to such exogenous agents as detergents or alcohol, the reaction of the enzymes obtained from lithium treated rats was different than that of control rats. These effects disappeared after ceasing lithium administration for one week. These data indicate changes in the structure of brain membranes after lithium administration. PMID- 6280733 TI - The effect of indomethacin and its ester on lysosomal enzyme release from polymorphonuclear leukocytes and intracellular levels of cAMP and cGMP after phagocytosis of urate crystals. PMID- 6280734 TI - Oropharyngeal Epstein-Barr virus excretion in rheumatoid arthritis. AB - We hypothesized that the defective cellular regulation of Epstein-Barr virus (EBV) in rheumatoid arthritis (RA) might be reflected in an increased rate of oropharyngeal virus excretion, but we found that the prevalence of excretion in 45 RA patients (22%) did not differ from 45 age- and sex-matched non-RA patients (24%) who were taking similar medications. Increased excretion rates correlated with corticosteroid therapy and male sex, but not with age or serum levels of EBV antibodies. PMID- 6280735 TI - [Enterocolitis caused by Shigella in an infant]. PMID- 6280736 TI - Lack of morphine-induced hyperactivity in C57BL/6 mice following striatal kainic acid lesions. AB - Bilateral injection of kainic acid (0.15 micrograms/0.3 microliters) into the striatum (caudatus/putamen) of C57BL/6 mice prevented stimulation of locomotor activity by morphine (20 mg/kg, i.p.). This effect was specific to morphine since mice with the same lesion did not show any impairment of amphetamine (2 mg/kg) induced locomotor hyperactivity. Histological inspections showed neuron damage also in the nucleus accumbens, while hippocampus was not damaged by kainic acid. Moreover, mice with kainic acid lesions in the hippocampus were more stimulated by morphine, compared with the morphine-injected sham lesion group. The results, which suggest the existence of non-catecholaminergic mediations in the locomotor effects of morphine, are discussed in terms of opioid systems in the brain. PMID- 6280737 TI - [Serological diagnosis of hepatitis A: determination with the RIA of anti-A specific IgM after absorption of IgG with protein A of Staphylococcus aureus]. AB - The research of specific IgM anti virus A (anti HAV-IgM), by absorbtion of IgG with Staphylococcus aureus A protein, has been conducted in the sera of 49 patients with acute hepatitis non B (HBsAg negative). Twenty-nine of these patients (59.2%) were anti HAV-IgM positive and therefore were diagnosed as viral hepatitis A, while twenty (40.8%) were anti HAV-IgM negative and were considered as non A-non B hepatitis. In the present work the practical utility of the detection specific IgM by absorbtion with A protein is discussed also in respect of the more expensive and less sensitive technique of gel fractionation of the sera. In addition our preliminary results document a decreasing incidence of viral hepatitis A in all the cases seen in the period 1978-1979. This event, which needs further investigations, could also be due to a more significant incidence of non A-non B hepatitis, probably as consequence of increasing drug addiction. PMID- 6280738 TI - [Demonstration of rotaviruses in feces by means of the complement fixation test]. AB - Complement fixation test for the detection of Rotavirus in stools has been evaluated. Removal of anticomplementary activity from the stools by absorption with complement or fetal calf serum was obtained. Then human reference serum pools were checked for the optimal dilution to be employed in the test. Sensibility and specificity of the c.f. method was determined on faecal specimens controlled by EM. The results indicate that c.f. test is adequate for, and easily adaptable to the detection of Rotaviruses in the stools. PMID- 6280739 TI - [Blood levels of catecholamines, cyclic AMP and cyclic GMP in endogenous adrenergic stimulation (cold pressor test and hand-grip) in humans]. PMID- 6280740 TI - RHC 3659: a new orally active angiotensin converting enzyme inhibitor in normal volunteers. AB - 1 The new converting enzyme inhibitor RHC 3659 was tested in 15 male volunteers. The study consisted of two parts: first, the ability of a single oral dose (5, 10, 20, 40 or 80 mg) to inhibit the pressor response to exogenous angiotensin I was tested with blood pressure and heart rate monitored continuously through an intraarterial catheter. A dose-related shift to the right of the pressor response curve to angiotensin I was observed with a peak occurring within 0.5 to 1 h. The pressor response to angiotensin II was unaffected. 2 In the second part, plasma renin and converting enzyme activity, angiotensin II and aldosterone were measured serially before and up to 8 h after administration of a single oral dose of RHC 3659. As expected. plasma angiotensin II and aldosterone fell within 30 min while plasma renin activity increased. Plasma converting enzyme activity was suppressed at 0.5 h in a dose-related manner with levels still below 30% of control 4 h following 80 mg of the inhibitor. 3 However, in vitro the enzyme inhibitor complex seemed quited fragile since during storage of the plasma samples at -20 degrees C, converting enzyme activity increased significantly already within days (P less than 0.001, n = 28) and continued to rise for more than 2 months. This fragility may explain the seemingly lower potency of RHC 3659 when compared to captopril. No side effects were observed. PMID- 6280741 TI - Biological effects of tremolite. AB - Tremolite is an amphibole which has been implicated in a variety of disease patterns in different parts of the world. It occurs in a number of phases, which are chemically identical but have specific physical characteristics. In an attempt to clarify the epidemiological findings, tremolite fibres of 3 specific forms--A, B and C--were characterized and studied for biological activity by: (i) in vivo intrapleural injection of rats (2 separate experiments--1 with poor survival). (ii) in vitro enzyme release from mouse peritoneal macrophages (iii) in vitro giant-cell formation in A549 cultures (iv) in vitro cytotoxicity for V79 4 cells. Sample C, which contained more long thin fibres than A and B, was alone in producing mesotheliomas. C, but not A or B, induced LDH and B-glucuronidase enzyme release, and induced giant cells. A was not cytotoxic, B moderately cytotoxic and C as highly cytotoxic as UICC crocidolite. The in vivo studies were marred by being split between 2 experiments, of which the second had poor survival. We are aware of the weakness of our in vivo data, but as Tremolite C was being considered for commercial use on the European market we felt it timely to submit our findings for publication. PMID- 6280742 TI - A comparison of the response of human lung carcinoma xenografts to vindesine and vincristine. PMID- 6280743 TI - The current status of systemic antifungal agents. PMID- 6280744 TI - Incidence of myeloperoxidase deficiency in an area of northern Italy: histochemical, biochemical and functional studies. AB - Forty-five subjects with a complete deficiency of myeloperoxidase were identified in an area of the region Friuli-Venezia Giulia in north-eastern Italy using the Hemalog D system as the screening technique. Histochemical and biochemical tests performed on the leucocytes of some of these subjects confirmed the defects shown by the Hemalog D system. The defect was of genetic origin in seven subjects. The genetic origin could be suspected in another eight subjects since more than two affected members were present in a given family. Eosinophil peroxidase, which is present in MPO deficient subjects, interfered with the guaiacol assay of MPO, and in several cases masked the genetic transmission. An assay was developed using o dianisidine as the electron donor which considerably reduced the interference by EPO. With this assay an autosomal recessive pattern of inheritance was found. The MPO deficient leucocytes had a higher respiratory burst than control cells and an impaired bactericidal activity, at early post-phagocytic periods, which became comparable to that of control cells at later stages. Particle ingestion by the MPO-deficient cells was normal. PMID- 6280745 TI - Receptors specific for thyroid hormones in nuclei of mammalian erythroid cells: involvement in erythroid cell proliferation. PMID- 6280746 TI - Unmasking moles. PMID- 6280747 TI - Immune electron microscopy and a cultural test in the diagnosis of adenovirus ocular infection. AB - Immune electron microscopy (IEM) and virus isolation in cell culture were compared in the diagnosis of adenovirus ocular infection during an outbreak of the disease in 1979. Eleven of 14 patients with a keratoconjunctivitis clinically indicative of adenovirus infection had IEM evidence of adenovirus infection or had the virus isolated from ocular swabs. The IEM was positive in 8 patients. Virus as isolated from 10 patients. IEM was positive in one culture negative patient. Since IEM provides a rapid and sensitive method for the detection of adenovirus in human tears, it may be a valuable diagnostic tool for the clinician. PMID- 6280748 TI - Formyl peptide chemoattractants: a model of the receptor on rabbit neutrophils. PMID- 6280749 TI - Regulation of the nuclear-coded peptides of yeast cytochrome c oxidase. AB - We have analyzed the catabolite regulation of cytochrome oxidase by assaying changes in the synthesis of precursors of the nuclear-coded peptides (IV--VII) of cytochrome c oxidase in an in vitro reticulocyte cell-free system programmed with RNA isolated from cells grown in either glucose or raffinose. As a first step, we have characterized antibodies which bind to the precursors of subunits V and VI. Initial translation products for subunits IV and VII have also been tentatively identified by utilizing these antibodies. The messenger RNAs coding for the precursors of the nuclear-coded subunits fall in the expected size range of 8--15 S. Catabolite repression of the nuclear-coded oxidase peptides appears to be regulated by the abundance of their messenger RNAs. Translation of messenger RNA isolated from yeast cells grown on glucose indicates a coordinate and uniform increase in precursor synthesis during glucose derepression. In contrast, when RNA isolated from raffinose (derepressed) grown cells is used to direct cell-free translation, precursor abundance is high throughout growth, although the synthesis of some of the species changes in a complex pattern of ratio and abundance. These data indicate that the abundance of the messengers for the nuclear-coded precursors is regulated in a fashion dependent on the physiologic state of the cell. PMID- 6280750 TI - Purification and characterization of the messenger ribonucleic acid capping enzyme GTP:RNA guanylyltransferase from wheat germ. AB - A GTP:RNA guanylyltransferase or capping enzyme has been purified approximately 2000-fold from wheat germ. The enzyme catalyzes the transfer of the GMP residue from GTP to the 5' end of RNA or synthetic polyribonucleotides. Diphosphate-ended polymers were capped more efficiently than molecules with triphosphate ends, and molecules with monophosphate ends were not capped at all. There appears to be little specificity since RNAs with purine or pyrimidine ends served as acceptors. Other features of the wheat germ RNA guanylyltransferase include relatively low Km values for GTP (2.7 microM) and ppA (pA)n (14.2 nM), a divalent cation requirement satisfied by low (0.5 mM) concentrations of MnCl2 or higher (5 mM) concentrations of MgCl2, and a pH optimum around neutrality. PMID- 6280751 TI - Binding of benzo[a]pyrene to histones and altered affinity of modified histone 1 for deoxyribonucleic acid. AB - The covalent binding of benzo[a]pyrene (B[a]P) to acid extractable chromosomal proteins and the subsequent effect on histone 1-DNA interaction have been characterized in a model system by utilizing calf thymus nuclei as targets and rat liver microsomes as an exogenous source of enzymes for the metabolic activation of B[a]P. A two-step ion-exchange chromatography and desalting procedure was employed for removing noncovalently bound B[a]P and other contaminants. Fluorography of acetic acid-urea and Triton-acetic acid-urea polyacrylamide gels indicated that H1 and H3 were the only principal histone targets in [3H]B[a]P-modified calf thymus nuclei. The validity of this assignment was confirmed by comparison of the chromatographic distributions of [3H]B[a]P cpm among peptides derived from the HClO4- soluble (H1) and HClO4-insoluble (core histones) protein fractions to the distributions obtained for authentic individual histone fractions. Comparison of amino acid compositions in individual peptide fractions which bound [3H]B[a]P differentially yielded some insight into the probable target amino acid residues for B[a]P binding. On the basis of electrophoresis in polyacrylamide gels, it appeared as if B[a]P had bound to multiple subfractions of H1 and H3. The equivalent distribution of covalently attached [3H]B[a]P among the major peptides of H1 and H3 modified either in intact nuclei or while free in solution implied that the relative accessibility of major portions of the H1 and H3 molecules for covalent B[a]P binding is not affected by interactions with DNA or other chromosomal proteins. Covalent attachment of [3H]B[a]P to purified H1 reduced the affinity of this histone for DNA-cellulose. PMID- 6280752 TI - Detection of separated amino proton resonance signals of adenine derivatives of low temperature and its application to estimation of population of the adenine uracil dimers in solution. AB - Splitting of the amino proton signals of 9-ethyladenine derivatives was found in proton nuclear magnetic resonance spectra at low temperature (ca. -30 degrees C). One of the separated signals corresponds to the syn amino proton relative to the N(1) nitrogen in the adenine ring and the other to the anti one. The phenomenon is ascribable to slowing down of the hindered rotation around the N(6)-C(6) bond, which has partial double bond character. On the addition of 1-cyclohexyluracil derivatives, one of the separated signals shifts downfield. From the analysis of the concentration dependence of the signals we could estimate the population of two kinds of adenine-uracil (AU) dimers that employ the syn and anti protons, respectively. i.e. the Watson-Crick-type and the Hoogsteen-type dimers. Independent of the substitution on the uracil ring, the Hoogsteen type is predominant at 70% and the Watson-Crick type at 30% (at -56 degrees C). On the other hand, with mixtures of general kinds of 9-ethyladenine derivatives with 1 cyclohexyluracil. the substituents on the adenine ring cause the population to deviate to extreme values; i.e., either the Watson-Crick-type or the Hoogsteen type dimer predominates. 2-Chloro-9-ethyladenine and N2-(dimethylamino)-9 ethyladenine take almost completely the Hoogsteen-type dimers, while 8-bromo-9 ethyladenine, N2-(methylamino)-9-ethyladenine, and 2-amino-9-ethylpurine predominant in the Watson-Crick-type dimers. PMID- 6280753 TI - Cytochrome c specific methylase from wheat germ. AB - The cytochromes c of plants (e.g., wheat germ) possess two trimethyllysines, residues 72 and 86. In order to investigate the nature of these methylations, we have purified a cytochrome c specific methylase S-adenosylmethionine: protein(lysine) N-methyltransferase (protein methylase III) from wheat germ 135 fold. The in vitro site of methylation by both the purified enzyme and crude wheat germ extract toward various forms of horse heart cytochrome c was localized by two dimensional peptide mapping, Aminex A-5 column peptide analysis, and CNBr cleavage analysis to be the residue 72 lysine. However, no additional sites, in particular residue 86, were seen to be methylated. Although the enzyme is highly specific toward cytochrome c as an in vitro protein substrate, avian cytochromes c are seen to be much better substrates than those from mammalian sources. The enzyme possesses an extremely low Km for apocytochrome c (1.21 microM), suggesting that methylation may occur before heme attachment in vivo. Various S adenosyl-L-homocysteine analogues were tested for their inhibitor capability toward the enzyme; it was observed that only the D and L forms of S adenosylhomocysteine are inhibitors while analogues modified in the adenine or homocysteine moieties do not possess inhibitory capability. Results from the Aminex A-5 column chromatography of horse heart cytochrome c chymotryptic digest showed the N epsilon-methyl-, N epsilon-dimethyl-m and N epsilon-trimethyllysine forms of the residue 68-74 peptide to elute earlier than the unmethylated form. This results suggest that the methylated peptides are less basic than the unmethylated form. PMID- 6280754 TI - Electrophoretic control of reconstituted adenine nucleotide translocation. AB - The initial velocity of adenine nucleotide exchange catalyzed by the reconstituted ADP-ATP carrier from beef heart mitochondria was measured under the influence of membrane potential and with different nucleotide distributions between the internal liposomal and the external buffer volume. Both Vmax and Km of adenine nucleotide uptake not only changed due to the applied potential but also depended on the respective nucleotide distribution. The rate equations for the ADP-ATP exchange under the various conditions were derived. These equations were simplified by assuming two alternative situations; either (a) af affinity type model, where the membrane potential influences only the affinity of the adenine nucleotide carrier toward ATP and ADP, or (b) a velocity type or distribution model, where the membrane potential modulates the rate constants of the ADP-ATP exchange. On the basis of several simplifications in the reconstituted system, the rate equations could be solved and the rate constants and dissociation constants of the exchange in the "energized" and in the "deenergized" state could be calculated. These values were used to derive prediction tables for normalized exchange rates under different nucleotide distributions, which were then compared with the experimental data. Only the exchange rates predicted by the velocity-type model agreed with the measured values. On the basis of this model a definite asymmetry caused by the membrane potential could be seen. Whereas this asymmetry is not very pronounced in the case of carrier-ADP complexes, about 40 times more ATP-loaded binding sites face the outside of the vesicles in the energized state. PMID- 6280755 TI - Stereochemistry of proton abstraction catalyzed by lysine and ornithine omega aminotransferases. AB - (6R)-L-[6-3H]Lysine and (6S)-L-[6]3H]lysine were prepared to investigate the stereochemical aspects of the reaction catalyzed by bacterial L-lysine epsilon aminotransferase. When (6R)-L-[6-3H]lysine was used as a substrate, the tritium label was retained in the product, delta 1-piperideine-6-carboxylate. In contrast, the radioactivity from (6S)-L-[6-3H]lysine was found in the solvent. Thus, the pro-S hydrogen at the prochiral C-6 carbon of L-lysine is specifically abstracted by L-lysine epsilon-aminotransferase. The proton exchange was observed by proton nuclear magnetic resonance analysis in the reaction of bacterial L ornithine delta-aminotransferase with L-ornithine in 2H2O. The isolated L-[5 2H]ornithine was converted to dextrorotatory 4-phthalimido[4-2H]butyrate. This indicates that L-ornithine delta-aminotransferase catalyzes the stereospecific exchange of the pro-S hydrogen at the prochiral C-5 carbon of L-ornithine with the solvent. PMID- 6280756 TI - Functional stability of Torpedo acetylcholine receptor. Effects of protease treatment. AB - The effect of tryptic degradation on structural and functional properties of the membrane-bound acetylcholine receptor from Torpedo californica has been investigated. Under conditions of proteolysis which resulted in extensive degradation of receptor subunits, the membrane preparations retained their full capability of mediating agonist-induced cation flux as measured in rapid kinetic experiments. Low concentrations on trypsin also cleaved receptor dimers to monomers, and this effect was paralleled by degradation of the Mr 65 000 subunits which are known to contain sulfhydryl group(s) involved in receptor dimerization through an interchain disulfide bond(s). This conversion to monomers occurred at lower trypsin concentrations when the enzyme was added to the outside of the vesicles compared with the effects observed when the enzyme was present inside the vesicles. Similarly Mr 43 000 protein consistently found in preparations of the membrane-bound acetylcholine receptor, which can readily be removed without apparent effect on receptor function, displayed greater susceptibility to proteolysis when trypsin was added to the exterior medium rather than inside the vesicles. The results emphasize the full functionality of the monomeric form of the acetylcholine receptor comprised of four polypeptides. PMID- 6280757 TI - Fluctuations and averaging of proton chemical shifts in the bovine pancreatic trypsin inhibitor. AB - The effects of motional averaging on the aromatic ring-current contribution to the proton chemical shifts in proteins are examined. Atomic trajectories obtained from a 96-ps molecular dynamics simulation of the bovine pancreatic trypsin inhibitor are used in conjunction with the Johnson-Bovey model of ring-current shifts to calculate the time evolution of the proton chemical shifts. Although large high-frequency fluctuations are observed (often greater than +/- 1 ppm), the average shift values in most cases are close to those obtained from the average structure; for some protons, significant differences are found. The calculated trends are used to probe the relationship between the average structure, atomic motions, and observed values of the proton chemical shifts. It is concluded that chemical shift values are in general most sensitive to the average structure of the protein and, because of the averaging involved, cannot be used directly to probe the short time structural fluctuations. PMID- 6280758 TI - Purification and properties of type 1 topoisomerase from chicken erythrocytes: mechanism of eukaryotic topoisomerase action. AB - A simple method for the purification of the major topoisomerase (topoisomerase 1) from chicken erythrocytes is described. Because of the generally repressed state of the chromatin from these nuclei, the heterogeneity of the non-histone proteins is reduced, and it is possible to purify this enzyme from a nuclear extract by a single chromatographic step. The chicken erythrocyte topoisomerase appears to be similar to previously described eukaryotic type I topoisomerases with respect to its physical and enzymological properties. The pattern of intermediate products generated during the action of chicken erythrocyte topoisomerase on a supercoiled closed circular DNA substrate has been examined quantitatively and has been shown to be consistent with a mechanism in which the enzyme closes its substrate DNA molecular after the removal of each superhelical turn and in which dissociation of the enzyme substrate complex may, but does not necessarily, occur after each cycle of the reaction. PMID- 6280759 TI - Interaction of cis-[Pt(NH3)2(H2O)2](NO3)2 with ribose deoxyribose diguanosine phosphates. AB - The three diguanosine phosphates GpG (4 X 10(-4) M), d(GpG) (10(-5) M), and d(pGpG) (10(-5) M) have been reacted with cis-[Pt(NH3)2(H2O)2](NO3)2 (1 Pt/dinucleotide) in water at pH 5.5 and 37 degrees C. In each case a single product is formed. The three complexes have been characterized by proton nuclear magnetic resonance (1H NMR) and circular dichroism (CD) analyses. They are N(7) N(7) chelates of the metal with an anti-anti configuration of the bases. They present a conformational change upon deprotonation of guanine N(1)H whose pKa is ca. 8.7 (D2O). Their CD spectra, compared to those of the free dinucleotides, exhibit an increase of ellipticity in the 275-nm region, which can be qualitatively related to the characteristic increase reported for platinated DNA and poly(dG) . poly(dC). These results are in favor of the hypothesis of intrastrand cross-linking of adjacent guanines, by the cis-PtII(NH3)2 moiety, after a local denaturation of DNA. PMID- 6280760 TI - Conformation of 2-[(deoxyguanosin-8-ylacetyl)amino]fluorene differs in protein free deoxyribonucleic acid and chromatin. PMID- 6280761 TI - Molecular topography of phytochrome as deduced from the tritium-exchange method. AB - The hydrogen-tritium-exchange measurements on phytochrome have been performed to detect the conformational differences between the red-absorbing (Pr) and the far red absorbing (Pfr) forms of phytochrome. The large and small Pfr molecules revealed more exchangeable protons that did the corresponding Pr molecules by 96 and 70 protons, respectively. These results suggest that the Pr leads to Pfr phototransformation is accompanied by an additional exposure of the peptide chains in the Pfr molecule. Of 1682 theoretically exchangeable hydrogens in undegraded phytochrome, only 442 (26%) and 346 (21%) protons were found to be exchangeable (excluding instantaneously exchangeable protons that cannot be determined by the present method). Thus, the phytochrome protein appears to be compact and highly folded. The kinetic analyses of the tritium exchange-out curves indicate that two kinetically different groups are responsible for the conformational differences between the Pr and Pfr forms of phytochrome. These components are due to (1) the exposure of hydrogen-bonded peptide segments (alpha helix and/or beta-pleated sheet) in the chromophore vicinity of Pfr and (2) the exposure of hydrogen-bonded peptide segments on the chromophore peptide domain as well as on the chromophore-free tryptic domain of undegraded phytochrome. PMID- 6280763 TI - Adenine nucleotide efflux in mitochondria induced by inorganic pyrophosphate. AB - The efflux of mitochondrial adenine nucleotide which is induced by addition of PPi to suspensions of rat liver mitochondria has been investigated. This efflux of adenine nucleotide is greatly stimulated by the uncoupler FCCP at 1 microM, Vmax being 6.7 nmol/min per mg protein as compared to 2.0 nmol/min per mg protein in its absence. The depletion process is inhibited by carboxyatractyloside. The Km for PPi of 1.25 mM is essentially unchanged when uncoupler is added. Quantitation of the individual adenine nucleotide species (ATP, ADP and AMP) and their relationship to the rate of efflux suggests that ADP is the predominant species being exchanged for PPi. PMID- 6280762 TI - Single-crystal electron paramagnetic resonance studies of photolyzed oxy- and nitric oxide-cobalt myoglobins. AB - Low-temperature photodissociation of oxygen from oxy-cobalt myoglobin was studied by single-crystal electron paramagnetic resonance (EPR) spectroscopy at 5 K. The photolyzed oxy-cobalt myoglobin exhibited an EPR spectrum consisting of two nonequivalent sets (species I and II) of the principal values and eigenvectors of the g tensors: g1I = 3.55, g2I = 3.47, and g3I = 2.26 for species I, and g1II = 2.04, g2II = 1.93, and g3II = 1.86 for species II, which resembled neither the deoxy nor the oxy form. Possible models of the photodissociated state of oxy cobalt myoglobin are proposed by comparison with cobalt porphyrin complexes. The photolyzed product of nitric oxide-cobalt myoglobin exhibited new EPR signals at g = 4.3 and a very broad signal at around g = 2. The principal g values have been determined from the single-crystal EPR measurements: g1 = 4.39, g2 = 4.27, and g3 = 4.00. Analysis of another EPR signal around g = 2 was difficult due to its broadness. Magnetic interactions were observed. An isotropic EPR signal at g = 4.3 suggested a weakly spin-coupled system between cobaltous spin (S = 1/2 or 3/2) and nitric oxide spin (S = 1/2). PMID- 6280764 TI - Anion and ionic strength effects upon the oxidation of cytochrome c by cytochrome c oxidase. AB - Citrate and other polyanion binding to ferricytochrome c partially blocks reduction by ascorbate, but at constant ionic strength the citrate-cytochrome c complex remains reducible; reduction by TMPD is unaffected. At a constant high ionic strength citrate inhibits the cytochrome c oxidase reaction competitively with respect to cytochrome c, indicating that ferrocytochrome c also binds citrate, and that the citrate-ferrocytochrome c complex is rejected by the binding site at high ionic strength. At lower ionic strengths, citrate and other polyanions change the kinetic pattern of ferrocytochrome c oxidation from first order towards zero-order, indicating preferential binding of the ferric species, followed by its exclusion from the binding site. The turnover at low cytochrome c concentrations is diminished by citrate but not the Km (apparent non-competitive inhibition) or the rate of cytochrome a reduction by bound cytochrome c. Small effects of anions are seen in direct measurements of binding to the primary site on the enzyme, and larger effects upon secondary site binding. It is concluded that anion-cytochrome c complexes may be catalytically competent but that the redox potentials and/or intramolecular behaviour of such complexes may be affected when enzyme-bound. Increasing ionic strength diminishes cytochrome c binding not only by decreasing the 'association' rate but also by increasing the 'dissociation' rate for bound cytochrome c converting the 'primary' (T) site at high salt concentrations into a site similar kinetically to the 'secondary' (L) site at low ionic strength. A finite Km of 170 microM at very high ionic strength indicates a ratio of K infinity m/K 0 M of about 5000. It is proposed that anions either modify the E10 of cytochrome C bound at the primary (T) site of that they perturb an equilibrium between two forms of bound c in favour of a less active form. PMID- 6280765 TI - Plasma membrane vesicles from isolated hepatocytes retain the increase of amino acid transport induced by dibutyryl cyclic AMP in intact cells. AB - We have investigated the effect of cyclic AMP on hepatic amino acid transport by measuring the uptake of L-alanine in plasma membrane vesicles purified from hepatocytes incubated without or with dibutyryl cyclic AMP. The application of an Na+ gradient to vesicles from hepatocytes exposed to dibutyryl cyclic AMP, compared to control, resulted in a greater transient accumulation of L-alanine, whereas in the presence of a K+ gradient a similar slow equilibration of L alanine was observed. Kinetic analysis of L-alanine influx revealed that the increased uptake resulted from an increased capacity (Vmax) with no change in the affinity (Km) of the carriers for L-alanine. These results strongly suggest that dibutyryl cyclic AMP induces stable changes at the membrane level probably by increasing the number of amino acid carrier molecules. PMID- 6280766 TI - Stimulatory (insulin-mimetic) and inhibitory (ouabain-like) action of vanadate on potassium uptake and cellular sodium and potassium in heart cells in culture. AB - (1) The influence of vanadate (Na3VO4) on sodium and potassium uptake as well as on cellular ion contents of sodium and potassium has been studied in heart muscle and non-muscle cells obtained from various species. An ouabain-like inhibition of potassium uptake (up to 50%), combined with a decrease of cellular potassium (up to 20%) has been observed by vanadate (10(-4)-10(-3) M) in heart non-muscle cells obtained from neonatal guinea pigs and chick embryos. In heart muscle and non muscle cells prepared from neonatal rats, as well as in Girardi human heart cells, a vanadate-induced stimulation of potassium uptake (up to 100%), combined with a rise in cellular potassium (up to 20%) and without significant alteration of cellular sodium, has been found. A slight increase of 22Na+ influx can be measured in rat heart muscle cells and in Girardi human heart cells in the presence of vanadate (10(-4)--10(-3) M). (2) In beating rat heart muscle cells in culture, detrimental effects of serum deprivation--concerning beating properties, potassium uptake and cellular potassium--can at least in part be overcome by addition of vanadate. Furthermore, this compound prevents ouabain-induced signs of toxicity (contractures) in these cells. (3) The stimulatory effects of vanadate on potassium can be mimicked by insulin (1-10 mU/ml). Furthermore, vanadate produces an insulin-like stimulation of 2-deoxy-D-glucose uptake in rat heart muscle and non-muscle cells as well as in Girardi human heart cells. (4) The experimental data demonstrate an ouabain-like inhibition as well as an insulin-mimetic stimulation of potassium-uptake in various heart cells. The reason for this antagonistic mode of action may be due to the different capabilities of the heart cell types to reduce vanadium in the V-valence state of vanadium in the IV-valence state, thereby favouring either ouabain-like inhibition (vanadium V) or insulin-mimetic stimulation (vanadium IV) of potassium transport. PMID- 6280767 TI - Inhibition of purified rabbit muscle phosphorylase a and phosphorylase b by polychlorinated biphenyls, polychlorinated biphenylols and polybrominated biphenyls. AB - Polychlorinated biphenyls, polychlorinated biphenylols and polybrominated biphenyls inhibited both rabbit muscle phosphorylase a and phosphorylase b (1,4 alpha-D-glucan:orthophosphate alpha-d-glucosyltransferase, EC 2.4.1.1). The degree of inhibition was dependent upon the relative hydrophobicity of the compounds and steric hinderance. 2,4,5,2',4',5'-Hexabromobiphenyl and Firemaster BP-6 were the most effective inhibitors (Ki, 15 . 10(-6) M). Phosphorylase b was inhibited by compounds of all three groups. 2,4,5,2',4',5'-Hexachlorobiphenyl and 2,4,5,2',4',5'-hexabromobiphenyl did not significantly inhibit phosphorylase a. All of the compounds inhibited phosphorylase a less than phosphorylase b, except 2',3',4',5,5'-pentachloro-2-biphenylol, which was equally effective on each enzyme. Kinetic analysis showed the inhibition was non-competitive and mixed. The results indicate that the compounds bind to hydrophobic site(s) on phosphorylase, access to which is limited by phosphorylation of serine 24. PMID- 6280768 TI - Interactions of fluoride and guanine nucleotides with thyroid adenylate cyclase. AB - The activation of bovine thyroid adenylate cyclase (ATP pyrophosphate-lyase (cyclizing), EC 4.6.1.1) by Gpp(NH)p has been studied using steady-state kinetic methods. This activation is complex and may be characterized by two Gpp(NH)p binding sites of different affinities with measured constants: Ka1 = 0.1 micro M and Ka2 = 2.9 micro M. GDP beta S does not completely inhibit the Gpp(NH)p activation: analysis of the data is consistent with a single GDP beta S inhibitory site which is competitive with the weaker Gpp(NH)p site. Guanine nucleotide effects upon F- activation of adenylate cyclase have been studied. When App(NH)p is the substrate, 10 micro M GTP along with 10 mM NaF gives higher activity than NaF alone, while GDP together with NaF inhibits the activity by 50% relative to NaF. These features are not observed when the complex is assayed with ATP in the presence of a nucleotide regenerating system or when analogs Gpp)NH)p or GDP beta S are used along with NaF. These effects were studied in three other membrane systems using App(NH)p as substrate: rat liver, rat ovary and turkey erythrocyte. No consistent pattern of guanine nucleotide effects upon fluoride activation could be observed in the different membrane preparations. Previous experiments showed that the size of soluble thyroid adenylate cyclase changed whether membranes were preincubated with Gpp(NH)p or NaF. This size change roughly corresponded to the molecular weight of the nucleotide regulatory protein. This finding, coupled with the present data, suggests that two guanine nucleotide binding sites may be involved in regulating thyroid cyclase and that these sites may be on different protein chains. PMID- 6280769 TI - The synthesis of deuterium-substituted, spin-labeled analogues of AMP and NAD+ and their use in ESR studies of lactate dehydrogenase. AB - Two spin-labeled analogues of AMP and NAD+ were synthesized, in which a perdeuterated nitroxide radical (4-amino-2,2,6,6-tetramethylpiperidine-1-oxyl, TEMPAMINE) was attached to C-6 or C-8 position of the adenine ring. The ESR spectra of these derivatives exhibit a 4-fold increase in sensitivity and a concomitant decrease in line-width as compared to the corresponding protonated analogues. The improved resolution of composite spectra consisting of freely tumbling and immobilized components is demonstrated in ternary complexes of the spin-labeled NAD+ derivatives with lactate dehydrogenase (L-lactate:NAD+ oxidoreductase, EC 1.1.1.27) and oxalate. PMID- 6280770 TI - Characterization of the soluble cyclic nucleotide phosphodiesterases in Xenopus laevis oocytes. Evidence for a calmodulin-dependent enzyme. AB - Cyclic nucleotide phosphodiesterase activities (3',5'-cyclic nucleotide 5' nucleotidohydrolase, EC 3.1.4.17) were found in the 40,000 X g supernatant fraction of homogenates of Xenopus laevis oocytes. In the supernatant, the ratio of the specific activity of cyclic AMP phosphodiesterase to that of cyclic GMP phosphodiesterase was 1.1 at the 1 micro substrate level. Two phosphodiesterase forms were isolated by centrifugation on sucrose gradient: a 3-4 S form hydrolyzing specificity cyclic AMP and a 6-7 S form hydrolyzing both cyclic nucleotides (cyclic AMP and cyclic GMP). The activity of the 6-7 S phosphodiesterase was characterized by its activation by 0.1 micro M calmodulin purified from beef pancreas in the presence of 50 micro M CA2+. The calmodulin dependence of this form was completely abolished in the presence of 1 mM ethyleneglycobis(beta-aminoethyl ether)-N-N,N',N'-tetraacetic acid (EGTA). Trifluoperazine at 0.1 mM inhibited both the freshly prepared crude enzyme and the partially purified 6-7 S form. On the other hand, no effect of cyclic GMP at 3 micro M was observed on cyclic AMP hydrolysis in the case of the supernatant or that of the partially purified phosphodiesterases. These data show the presence of a calmodulin-dependent phosphodiesterase in the soluble fraction of X. laevis oocytes. PMID- 6280771 TI - The role of endogenous phosphatidylcholine and ceramide in the biosynthesis of sphingomyelin in mouse fibroblasts. AB - The intracellular location of sphingomyelin formation via the cholinephosphotransferase reaction from both endogenous an exogenous phosphatidylcholine and ceramide substrates has been studied in the subcellular membrane fractions prepared from mouse fibroblasts. The enzyme was found to be located in both the plasma membrane and the Golgi fractions. Activity in the Golgi fraction was stimulated to a greater extent by the addition of exogenous ceramide than was the activity in the plasma membrane fraction. It is concluded that endogenous phosphatidylcholine is available to the cholinephosphotransferase at saturating concentration and, therefore, is not rate-limiting. In contrast, the very low concentration of endogenous ceramide seems to limit the reaction rate, necessitating supplementation with exogenous material Both endogenous substrates are shown to be utilized in an intramembranous rather than an intermembranous reaction. The capacity of the plasma membrane fraction to synthesize sphingomyelin from endogenous phosphatidylcholine and ceramide was found to be sufficiently high to account for the rate of net synthesis of plasma membrane-bound sphingomyelin observed in the logarithmically multiplying cell culture. In contrast, the Golgi fraction displayed only 26% of the expected capacity, but it was stimulated 6-fold by the addition of exogenous ceramide. These results demonstrate that the total cellular sphingomyelin of the mouse fibroblasts can be provided via the cholinephosphotransferase pathways and that the plasma membrane and the Golgi fraction are most probably the intracellular sites of sphingomyelin biosynthesis. PMID- 6280772 TI - Metabolism of phosphoinositides in the rat erythrocyte membrane. A reappraisal of the effect of magnesium on the 32P incorporation into polyphosphoinositides. AB - The metabolism of phosphoinositides was investigated in the red blood cell membrane of the rat by measuring 32P-incorporation into phospholipids after incubation of membranes with [gamma-32P]ATP in a medium containing magnesium. A new chromatographic procedure has been developed which facilitates the separation of triphosphoinositide, diphosphoinositide and phosphatidylinositol from the phospholipids present in lipid extracts of incubated 'ghost' under our experimental conditions only two phospholipids, diphosphoinositide and triphosphoinositide, were 32P-labelled. Furthermore, the results indicate that either di-or triphosphoinositide could be labelled preferentially, depending upon the magnesium concentration of the incubation medium. This clarifies some apparent discrepancies reported in the literature between the 32 P labelling of polyphosphoinositides observed in intact erythrocytes and that observed with 'ghost' membranes. In addition, the enzymatic pathways involved in the phosphoinositide metabolism are discussed. PMID- 6280773 TI - Chronic and acute effects of thyrotropin on phosphatidylinositol turnover in cultured porcine thyroid cells. AB - The 32P incorporation into phospholipids of isolated porcine thyroid cells, cultured for 1-4 days, has been studied in subsequent 2-h incubations. Along with culture ageing, decreased 32P incorporation into total phospholipid of control cells was observed. The presence of 40 munits/ml TSH during the 2 h incubation yielded a relative increase in labelling of phosphatidylinositol, named 'acute phospholipid effect'. A chronic treatment of the cells with TSH concentration ranging from 0.1 to 10 munits/ml ensured the maintenance of a high turnover rate of total phospholipids. The analysis of individual phospholipids revealed that 1 day culture cells in the presence of 0.1 munits/ml TSH presented a strong increase of phosphatidylinositol labelling. This 'chronic phospholipid effect' of TSH can be reproduced by a chronic treatment with dibutyryl cyclic AMP (10(-3)M) or prostaglandin E2 (10(-6)M), which did not evoke a classical phospholipid effect in a 2 h incubation. If TSH (40 munits/ml) is added to the cells in a 2 h incubation, control cells show the classical phospholipid effect whereas cells chronically treated with TSH, dibutyryl cyclic AMP or prostaglandin E2 presented a 'reverse phospholipid effect' i.e. a relative decrease in phosphatidylinositol labelling. 10(-4)M cycloheximide presence during the last 12 h of culture prevented the establishment of the 'chronic phospholipid effect' and of its consequence, 'the reverse phospholipid effect'. On the basis of these results a scheme is proposed in keeping with current hypotheses concerning phosphatidylinositol metabolism. PMID- 6280774 TI - Enhanced production of hydroxyl radicals by the xanthine-xanthine oxidase reaction in the presence of lactoferrin. AB - The generation of hydroxyl radicals by the xanthine-xanthine oxidase reaction (C. Beauchamp and I. Fridovich (1970) J. Biol. Chem. 245, 4641-1616) has been shown to be increased by iron-saturated lactoferrin isolated from pig neutrophils. Hydroxyl radical production, measured by EPR spin trapping and by ethylene production from alpha-keto-gamma-methiol butyric acid, has been demonstrated to be produced by a Fenton-type Haber-Weiss reaction catalysed by lactoferrin. The possibility that lactoferrin catalyses such a reaction in vivo is considered. PMID- 6280775 TI - Relevance of ferritin-binding sites on isolated mitochondria to the mobilization of iron from ferritin. AB - Iron can be released from ferritin and utilized by isolated rat liver mitochondria for the synthesis of heme. Mobilization of iron from ferritin is initiated by the binding of ferritin to the mitochondria in an manner compatible with binding sites or receptors for ferritin on the mitochondria. The binding completes rapidly, it is independent of temperature, saturable, reversible and enhanced by K+ and Mg2+. The amount of ferritin binding sites is approx. 0.8 pmol/mg mitochondrial protein, and the affinity constant is 6.4 . 10(6)M-1. The binding kinetics correlate well with the functional features of the ferritin mitochondrial interaction: i.e. mobilization of iron from ferritin followed by insertion of the iron into heme. The results support the concept of ferritin as a possible donor of iron to the mitochondria. PMID- 6280776 TI - Correlation of the biosynthesis of prostaglandin and cyclic AMP in monolayer cultures of rabbit articular chondrocytes. AB - We have utilized ionophores to test whether stimulation of chondrocyte prostaglandin biosynthesis is accompanied by an increase in cyclic nucleotide levels in these cells. Radioimmunoassay of prostaglandin E2, 6-oxo-prostaglandin F1 alpha (the stable metabolite of prostaglandin I2) and prostaglandin F2 alpha showed that synthesis of each was stimulated by the divalent-cation ionophore, A23187 after short-term incubation (1-7 min) in serum-free medium. No stimulation of thromboxane B2 was detected. Two monovalent ionophores, lasalocid and monensin failed to stimulate prostaglandin biosynthesis after short-term incubation. Ionophore A23187-stimulated prostaglandin biosynthesis was variably and partially inhibited by sodium meclofenamate, indomethacin and aspirin, but not by sodium salicylate. Ionophore A23187-stimulated prostaglandin biosynthesis was accompanied by a 7.5-fold increase in cyclic AMP levels after 15 min. Sodium meclofenamate, indomethacin and aspirin which inhibited prostaglandin E2 biosynthesis also reduced cyclic AMP levels. Exogenous prostaglandin E2 (1 microgram/ml) stimulated cyclic AMP biosynthesis, which was not inhibited by aspirin. These results indicated that prostaglandins can be considered as one of the local effectors controlling cyclic AMP production in articular cartilage. PMID- 6280777 TI - Human platelet activation by an alkylacetyl analogue of phosphatidylcholine. PMID- 6280778 TI - Insulin and dexamethasone effects on liver fructose bisphosphatase. AB - Insuling perfusion of isolated diabetic rabbit liver and glucocorticoid perfusion of isolated normal rabbit liver resulted in an increased accumulation of immunoreactive fructose bisphosphatase (nmol/g). The insulin effect was maximal at 30 min with concentration (nmol/g) and specific activity (units/nmol) returning to normal within 2 h after insulin removal. Similarly, dexamethasone perfusion increased enzyme concentration and decreased its specific activity; however, the effects were maximal at 60-120 min. The results parallel those previously demonstrated in whole animals and establish that both hormones influence the enzyme by direct effects on the liver. PMID- 6280779 TI - [Mechanical destruction of collagen]. AB - Mechanical destruction of collagen was studied by means of electrophoresis in polyacrylamid gel and ESR-spectroscopy. It has been stated that the rates of the breakage of polypeptide chains measured by these two methods are in a good agreement. An analysis of ESR spectra of free radicals in collagen suggests that mechanical action induces the rupture of C alpha-N-bonds. PMID- 6280780 TI - [Change of current-voltage characteristics of amphotericin B ion channels accompanying its incorporation into the membrane]. AB - Non-linearity of amphotericin B current-voltage characteristics is determined by means of the third harmonic generated in the membrane. A change of the sign of non-linearity is revealed during incorporation of antibiotic molecules into a membrane, i. e. the incorporation is not a simple increase of the number of similar ionic channels in the membrane. PMID- 6280781 TI - [Effect of pH and substitution of H2O for D2O on oxygen liberation by chloroplasts]. AB - The rate of dark relaxation of the oxygen evolving system in chloroplasts is shown to depend on the value of the surface charge of some chloroplast membrane component having protein nature and isoelectric point at pH 6.0. The substitution of H2O for D2O leads to isoelectric point shift of this protein. PMID- 6280782 TI - [Low potential c-type cytochrome of Thiocapsa roseopersicina]. AB - The low potential c-type cytochrome from the phototrophic purple sulphur bacterium Thiocapsa roseopersicina, strain BBS was isolated in electrophoretically homogeneous state. The bulk of the cytochrome (approximately 90%) after disruption of the cells remained in the membrane fraction. The absorption spectrum of the cytochrome was characterized by the maxima at 420, 523 and 552 nm in the reduced state and at 408 nm in the oxidized one. The cytochrome interacted with CO in the reduced state. The molecular weight of the cytochrome is 50 000. The cytochrome contains great amounts of phenylalanine, leucine, valine, aspartic and glutamic acids and can be reduced by dithionite but not by cysteine, sulfide or ascorbate. Besides, the cytochrome can also be reduced by NAD(P)H in the presence of NAD(P)-reductases of T. roseopersicina, when ferredoxin of Spirulina platensis or benzyl viologen are added to the reaction mixture. The cytochrome can act as an electron donor (acceptor) for T. roseopersicina hydrogenase. PMID- 6280783 TI - Comparison of Nernst-Planck and reaction rate models for multiply occupied channels. AB - The Nernst-Planck continuum equation for a channel that can be occupied by at most two ions is solved for two different physical cases. The first case is for the assumption that the water and ion cannot get around each other anywhere in the channel, so that if there are two ions in the channel the distance between them is fixed by the number of water molecules between them. The second case is for the assumption that there are regions at he ends of the channel where the ions and water can get around each other. For these two cases, the validity of the simple two-site reaction-rate approximation when there is a continuously varying central energy barrier was evaluated by comparing it with the exact Nernst-Planck solution. For the first continuum case, the kinetics for the continuum and reaction-rate models are nearly identical. For the second case, the agreement depends on the strength of the ion-ion interaction energy. For a low interaction energy (large channel diameter) a high ion concentrations, there is a large difference in the flux as a function of voltage for the two models-with the continuum flux becoming more than four times larger at 250 mV. Simple analytical expressions are derived for the two-ion continuum channel for the case where the ends are in equilibrium with the bulk solution and for the case where ion mobility becomes zero when there are two ions in the channel. The implications of these results for biological channels are discussed. PMID- 6280785 TI - Delayed kinetics of squid axon potassium channels do not always superpose after time translation. AB - The activation of potassium ion conductance in squid axons by voltage-clamp depolarization is delayed when the depolarizing step is preceded by a conditioning hyperpolarization of the axonal membrane. Moreover, the control conductance kinetics superpose with the delayed kinetics when they are translated along the time axis by an amount equal to the delay. We have found that the degree of superposition with internally perfused axons depends upon voltage-clamp protocol. The kinetics superpose almost exactly for modest test depolarizations, whereas they clearly fail to superpose completely for more positive levels of membrane depolarization. We have modeled these results by incorporating a time dependence into the rate constant of activation of potassium channel gates in the Hodgkin and Huxley model of potassium ionic conductance. PMID- 6280784 TI - Lipid-protein interactions in sarcoplasmic reticulum are not perturbed by ionophore A23187. An EPR and fluorescence study. AB - The divalent-cation ionophore A23187 at micromolar concentrations prevents the ATP-dependent accumulation of calcium into sarcoplasmic reticulum vesicles. Under the same conditions and throughout the temperature range of 4 degrees-37 degrees C, A23187 has no effect on either the rotational motion of the Ca2+ -ATPase in the membrane, or on the mobility of the lipid acyl chains. The steady-state fluorescence polarization of a polyene fluorescent probe incorporated into the membrane lipids was similarly unaffected by the ionophore. These results show conclusively that the mechanism of action of the ionophore does not involve significant of lipid-protein interactions. PMID- 6280786 TI - In vitro processing of the adenosine analog formycin A to the mono-, di-, and triphosphate by a soluble multienzyme system from mouse liver. AB - The adenosine kinase activity present in a soluble preparation from rat liver was investigated using formycin A (FoA), a fluorescent analog of adenosine as the phosphoryl acceptor and ATP as the donor. Reversed-phase high-performance liquid chromatography (h.p.l.c.) was used to separate substrate from product, and the progress of the phosphorylation reaction was followed by monitoring fluorometrically the amount of formycin 5'-monophosphate (FoMP), and the AMP analog, that was formed. The results showed that while FoMP was formed during the reaction indicating that an adenosine kinase activity was present, both formycin 5'-di- and triphosphate (FoDP and FoTP respectively), the corresponding analogs of ADP and ATP, were also formed, suggesting than an adenylate kinase activity was present. This result was confirmed with FoMP as the substrate and showing the formation of FoDP and FoTP. Other experiments carried out with FoMP as the substrate revealed the formation of FoA. Taken together, these results indicated that a 5'-nucleotidase activity as well as an adenylate kinase was present. Using this analog and h.p.l.c., it has been possible to demonstrate for the first time in an in vitro system the complete salvage of a nucleoside to the triphosphate level. PMID- 6280787 TI - Arachidonate-induced fibrinogen binding to thrombin-degranulated rabbit platelets is independent of released ADP. PMID- 6280788 TI - Demonstration of a putative adenosine receptor in rabbit aorta. AB - This study demonstrates the existence of adenosine receptor(s) in a homogeneous population of cultured smooth muscle cells and intact muscle strips from rabbit aorta. Binding assays with 3H-adenosine were performed using the crude microsomal fraction of the cultured cells. Adenosine binding was reversible and displayed a single species of saturable binding sites. The nuclear and soluble fractions showed negligible binding. Adenosine uptake blockers (e.g., nitrobenzylthioinosine, dipyridamole) had no effect on its binding. Theophylline inhibited the binding and the relaxation of aortic strips to adenosine. The data suggest the existence of specific binding sites for adenosine on the surface of aortic smooth muscle cells. PMID- 6280789 TI - [Small lung cancer]. PMID- 6280791 TI - [Ultrastructural study of small cell carcinoma (author's transl)]. AB - Fifty bronchopulmonary tumors were studied by light and electron microscopy: 39 carcinomas (25 small cell carcinomas, 8 undifferentiated carcinomas, 6 poorly differentiated squamous cell carcinomas) and 11 carcinoid tumors (5 central, 4 peripheral, 1 atypical, 1 of the tumorlet type). The results indicate that small cell carcinoma is a precise entity characterized by the presence of neurosecretory granules in the cytoplasm. Ultrastructurally carcinoid tumor resemble small cell carcinomas and there is a continuous spectrum of differentiation between the less undifferentiated neuro-endocrine tumors, i.e. the small cell anaplastic carcinomas, and the higher differentiated types, i.e. the carcinoids. The diagnostic and prognostic value of the ultrastructural findings are discussed. PMID- 6280790 TI - [Small cell carcinoma. Incidence, histopathology and anatomical features. Analysis of 465 autopsied bronchopulmonary carcinomas (author's transl)]. AB - 465 patients with broncho-pulmonary malignant tumors have been autopsied. Small cell carcinoma was diagnosed in 22.5 per cent of these patients. The histo cytological variants of these tumors (lymphocytoid, polygonal, fusiform and polymorphic) had the same general characteristics (age, sex, survival) and a similar clinical course. Grossly and histologically, the bronchial tumor, always located in proximal bronchial tree, largely involved the mediastinum. Metastases were peculiarly frequent to the liver (69%), to bone (64.2%) and to the central nervous system (36.2%). Three Schwartz-Bartter syndromes and two Denny Brown sensitive neuropathies were noted in this statistical study. PMID- 6280792 TI - [Undifferentiated small cell carcinoma and diffuse endocrine system (author's transl)]. AB - Oat cell carcinomas of the lung are sometimes associated with endocrine secretion. Resulting syndromes, first regarded as "paraneoplasic", are in fact imputable to secretory activity of the tumoral cells, as demonstrated by electron microscopy and immunofluorescence studies. Such ascertainments are explained by the existence in the bronchial tree of a diffuse endocrine system sector, from which carcinoid and oat cell carcinoma originate. However, the authors consider other histogenetic hypotheses and rapidly survey the extra-pulmonary oat cell carcinomas. PMID- 6280793 TI - Endoscopic diagnosis of small anaplastic carcinoma: difficulties and means of determining contour. AB - Small cell carcinoma do not have specific features, because bronchi can present either with a tumoral obstruction (42%) or with an infiltration (41%) or a compression (14%) or normal (3%) pattern. The improvement of the endoscopic technology, the more frequent changes in biopsy forceps, the increasing ways in performing these biopsies are expected to decrease the number of false negative results. But, if were the case, more sophisticated diagnosis tests under fluoroscopy are needed. The transthoracic needle aspiration-biopsy because of its better sensitivity and less fatal complications seems to be preferred to the transbronchial biopsy. Using such approach it can be expected that small cell carcinoma, which chimio sensitivity is known, will be diagnosed accurately. PMID- 6280794 TI - Small cell anaplastic carcinoma of the lung. The Cancer and Leukemia Group B Experience. AB - Three protocols have been evaluated by the Cancer and Leukemia Group B (CALGB) for the treatment of small cell anaplastic cancer of the lung between 1972 to 1980. The first protocol (CALBG 7283) was a randomized comparison between 4 arms: high dose cyclophosphamide alone, cyclophosphamide plus methotrexate, cyclophosphamide plus methotrexate plus vincristine, and cyclophosphamide plus high-dose methotrexate plus vincristine. Response rate and survival were not significantly different among the 4 arms. Effectiveness of prophylactic brain radiotherapy and usefulness of maintenance chemotherapy were demonstrated. The next protocols (CALGB 7781 and 7782) compared two adriamycin containing regimens: MACC (methotrexate, adriamycin, cyclophosphamide, CCNU) versus CCV/AV (cyclophosphamide, CCNU, vincristine, alternating with adriamycin and vincristine). Split course radiotherapy (total 45 Gy over 4 weeks) was also given to all patients with limited disease (CALGB 7781) as well as prophylactic brian irradiation. Radiotherapy to the chest was also given in an additional arm of CALGB 7782 for patients with extensive disease. No significant differences were seen for response and survival within categories defined by extent of disease. Overall about 25 per cent of patients achieving complete remission remain alive at 2 years. The prognostic impact of different factors including extent of disease, performance of status, age and sex are discussed. PMID- 6280795 TI - Experience of the National Cancer Institute (USA) in the treatment and biology of small cell lung cancer. AB - This retrospective study deals with 168 patients with small cell lung cancer (SCLC) who were treated at the NCI March 1973 and July 1977 with intensive chemotherapy. Results showed that 12 per cent of all the patients were disease free for more than 40 months (25% of the limited disease and 3% of the extended stage disease patients which were treated. Complete remission was essential to prolonged survival. The development of a complete remission was favored by a satisfactory initial general condition, localised disease or presence of only one site of metastatic disease. Initial treatment included the following combinations: cyclophosphamide, methotrexate and CCNU. High doses are necessary. However, above a certain level, the toxicity increased without a corresponding increment in activity. Addition of another association without cross-resistance (vincristine-adriamycine-natulan or VP-16 and isophosphamide) may induce a complete remission in cases where only a partial remission was obtained initially. In limited disease, radiotherapy seemed to prolong disease-free survival. This lead the authors to study the use of initial radiotherapy, at the rate of 40 grays in 3 weeks in 15 fractions, at the same time as chemotherapy. The radiotherapist must take certain precautions in the use of this double treatment: reshaped fields and insertion of a spinal cord block above 2 000 rads. In extended disease, thoracic irradiation seemed to be of no benefit. Pilot studies are not underway using intensive chemotherapy with multiple tumor site irradiation and autologous marrow implants 48 per cent of all patients with SCLC will develop cerebral metastatic disease (44% intracranial, 13% leptomeningeal, 9% epidural). At 30 months, 75 percent of those who did not receive prophylactic irradiation developed cerebral disease whereas only 40 per cent of the patients who received a prophylactic dose of 30 grays developed cerebral disease. Inspite of the indisputable but incomplete local benefit of prophylactic brain irradiation, it did not prolong survival. Important biological studies are underway at the NCI. - Cell clone cultures have been established. These were enhanced by the addition of arginine vasopressive (AVP) and bombesine. These two substances may be considered as markers of APUD system and are secreted by the SCLC. Bombesine has been isolated in all the cultures tested. This possible marker was perhaps responsible for the anorexia and cachexia in these patients. - These cultures have allowed for identification of deletion 3p (14-23) chromosome anomalies in all the samples examined. - In vitro chemotherapy studies of these cultures have been carried out. Their correlation with clinical results were encouraging (100% negative p/n; 75% positive p/n). - Lastly, monoclonal AC have been prepared. Inspite of their imperfect specificity and heterogeneity with regard to tumor cells, their potential advantages were considerable. PMID- 6280796 TI - Combination of surgery and polychemotherapy for cure in early small-cell bronchial carcinoma. AB - In 1969 a cooperative randomized two arm study of surgery and adjuvant chemotherapy was started. Within three years after the operation the polychemotherapy was given intermittently using a combination of cyclophosphamid + 5-FU + MTX + vinblastin. From a total of 859 patients operated for cure preliminary evaluations per 1.3.1981 found 53 patients with small cell type carcinomas. An increase of the 3-year survival rate of 13 per cent shows a clear positive therapeutic effect. This also demonstrates the importance of surgery even on small cell bronchogenic carcinoma by decreasing the tumor burden. To evaluate wether or not the number of long-term survivors can be increased furthermore, a new randomized trial was started in 1979 to compare the polychemotherapy used with a new designed intermittent polychemotherapy-schedule using alternating cyclophosphamid + CCNU + MTX, there after cyclophosphamid + adriamycin + vincristin and there after ifosfamid + VP-16; then radiation therapy and there after repeating the whole sequence. PMID- 6280797 TI - [Sequential chemotherapy and radiotherapy modalities in small cell lung carcinoma. Preliminary results]. AB - Two non randomized therapeutic trials have been conducted in oat cell carcinoma of the lung with limited disease. The first protocol combined chemotherapy (adriamycin, cyclophosphamide, VM 26, procarbazine) with mediastinal and supra clavicular radiotherapy in classical fractionation (40 grays in 4 weeks). The second combined a more intensive chemotherapy (adriamycin, VP 16, methotrexate, cyclophosphamide) with an alternating radiotherapy (3 series of 15 grays). All patients received prophylactic brain irradiation. Preliminary results show a 9 month survival rate of 34 per cent and 62.5 per cent respectively. In addition, all patients in the second protocol presented a complete response at the end of the induction treatment. These preliminary data encourage proceeding with this kind of combined treatment. PMID- 6280798 TI - An asymmetrical kinetic model for veratridine interactions with sodium channels in molluscan neurons. PMID- 6280801 TI - Endocrine pancreatic carcinoma and syndrome of inappropriate secretion of antidiuretic hormone. PMID- 6280799 TI - The association between the melanocyte-stimulating hormone receptor and the alpha 2-adrenoceptor on the Anolis melanophore. AB - 1 The primary effect of catecholamines was to lighten Anolis skin previously darkened by alpha-melanocyte-stimulating hormone (alpha-MSH). In concentrations above 10(-7) M noradrenaline, 10(-6) M adrenaline and 10(-5) dopamine, darkening of subpopulations of melanophores occurred. Subsequent experiments were concerned with the effect of low catecholamine concentrations on alpha-MSH action. 2 The relationship between MSH receptors and alpha-adrenoceptors on the Anolis melanophore was studied by a kinetic approach using the rate bioassay method and by use of alpha-adrenoceptor agonists and antagonists. 3 alpha-MSH dose-response curves were shifted, in parallel, to the right in the presence of the catecholamines, noradrenaline, adrenaline and dopamine, and Lineweaver-Burke plots and Arunlakshana-Schild plots indicated that the catecholamines antagonized MSH action by a competitive mechanism. 4 Phentolamine had an inhibitory effect on the action of adrenaline but not on the action of MSH. Therefore MSH and catecholamine actions were mediated by separate receptors. 5 The classical kinetics of competition are not confined to competition at a single receptor. 6 The alpha-adrenoceptor was defined as the alpha 2-subtype since (a) the alpha 2 selective agonist, clonidine, was found to mimic catecholamine action. (b) The alpha 2-selective antagonist, yohimbine, blocked the actions of clonidine and adrenaline. (c) The alpha 1-selective antagonist, prazosin, had negligible blocking effects on adrenaline and clonidine. 7 We conclude that a close association exists between the separate MSH receptor and alpha 2-adrenoceptor on the Anolis melanophore. The competition that takes place between MSH and catecholamines must occur after hormone-receptor interaction, possibly through a common adenylate cyclase moiety oppositely controlled by the two receptors involved. PMID- 6280800 TI - Synthesis of new haloperidol analogues and characterization of their interactions with alpha-adrenoceptors in rat parotid slices and human platelet membranes. AB - 1 The synthesis of several butyrophenone analogues of haloperidol is described. 2 The effects of these compounds on alpha-adrenoceptors were evaluated by examining their ability to reduce alpha 1-stimulated K+ release from rat parotid slices and to displace [3H]-phentolamine from human platelet membrane alpha 2-adrenoceptors. 3 The affinity of haloperidol and its analogues for alpha 1-receptors was found to be 1 to 2 orders of magnitude greater than that for alpha 2-adrenoceptors. These observations suggest that most of the alpha-adrenoceptor activity of butyrophenones results from their interaction with alpha 1-adrenoceptors. 4 The relatively high affinity of the butyrophenones for alpha 1-adrenoceptors suggests that they may be useful as probes in studies of alpha 1-adrenoceptors in these and other tissues. PMID- 6280802 TI - Is bran useful in diverticular disease? PMID- 6280803 TI - Characterization of adenosine receptors in brain using N6 cyclohexyl [3H]adenosine. AB - The presence of distinct binding sites for adenosine in both the CNS and PNS has been proposed in numerous studies. The recent availability of stable adenosine analogues such as cyclohexyladenosine, 2-chloroadenosine and diethylphenylxanthine has made the characterization of such a receptor feasible. In the present report the binding of N6 cyclohexyl [3H]adenosine ([3H]CHA) to rat brain synaptosomal membranes is characterized. [3H]CHA binding is saturable and exhibits a biphasic kinetic saturation profile characteristic of 2 binding sites. The high affinity site has a Kd of 0.7 nM and the low affinity site 2.4 nM. The respective Bmax values are 230 and 120 fmol/mg protein in fat forebrain. The highest density of binding sites is found in the hippocampus and subcellular distribution studies indicate that the [3H]CHA site is predominantly synaptosomal. [3H]CHA binding is highly dependent in the presence of adenosine deaminase since only 30% of the binding capacity is observed in synaptosomal membranes not treated with this enzyme. Of the many cations and anions tested only copper and zinc have effects on [3H]CHA binding. Both metals are potent inhibitors of binding with copper having an IC50 of 30 microM and zinc 150 microM. Sulfhydryl reducing and alkylating agents also inhibit binding indicating that the binding site is a sulfhydryl-dependent protein. PMID- 6280804 TI - CSF beta-endorphin-immunoreactivity in normal, schizophrenic, depressed, manic and anorexic subjects. AB - beta-Endorphin immunoreactivity was measured in cerebrospinal fluid of 75 medication-free subjects: normal, depressed, schizophrenic, and anorexic. No significant differences in beta-endorphin immunoreactivity were found. Affinity extraction chromatography revealed beta-lipotropin and beta-endorphin, but no apparent precursors. PMID- 6280805 TI - Indirect ultrastructural evidence for lithium uptake by cultured rat cerebral cells through the sodium channel. AB - Cerebral cortices, prepared from 18-day-old rat embryos, were grown in explant cultures for 18 days. They were then incubated for 25 min in media of 3 types of cationic composition with and without tetrodotoxin and/or veratridine. Qualitative and quantitative ultrastructural observations revealed a considerable swelling of neuronal elements following veratridine exposure in both Na+ and Li+ media; this swelling was prevented by tetrodotoxin. By contrast, veratridine failed to induce swelling of neuronal profiles in a choline+ medium. The results indirectly indicate that Li+ ions enter cultured rat cerebral cells through the Na+ channels. PMID- 6280806 TI - Development of methionine-enkephalin and naloxone binding sites in regions of rat brain. AB - The development of opiate receptors in whole forebrain, brain stem and cerebellum of developing rats was measured by specific [3H]methionine-enkephalin ([3H]Met Enk) and [3H]naloxone binding. The level of opiate receptor as determined by the binding of these two opiates varied with brain region as well as with age; however, the amount of [3H]naloxone bound in the same brain region obtained from animals of the same age was greater than that of [3H]Met-Enk. Kinetic analyses of these two opiate binding sites in newborns of the same age revealed that they differed in their apparent affinities and receptor numbers. Moreover, both morphine and naloxone were equally active in competing for [3H]Met-Enk binding in all 3 brain regions during the first month after birth, whereas methionine enkephalin was only about 30-50% as active as morphine n competing for [3H]naloxone binding in that period. These results suggest that in the rat brain there are two types of opiate receptors and that the heterogeneity of opiate receptors is already apparent during early postnatal life. PMID- 6280807 TI - Inhibition of growth of mouse neuroblastoma cels by protein factor derived from rat glioblasts. AB - Growth inhibitory factor for mouse neuroblastoma cells was detected in the culture medium of fetal rat glioblasts, and was partially purified and characterized. The factor had an apparent molecular weight of 75,000 and an isoelectric point of 5.8, and showed no esterase activity. It possessed the activity to promote morphological differentiation including the formation of neural processes and the inhibition of cell division when tested on mouse neuroblastoma cell lines (Neuro 2a, NS-20Y, and NIE-115). The activity was susceptible to protease digestion and heat treatment. The serum over 25% cancelled the inhibitory activity of this factor. The factor failed to increase the intracellular contents of cAMP and cGMP. It showed no effect on either morphological differentiation or proliferation of glial cell lines, suggesting that under physiological conditions the factor acts specifically on neuronal cells. PMID- 6280808 TI - Opiate receptor development in midbrain and forebrain of posthatch chicks. AB - Specific binding [3H]naloxone to midbrain and forebrain was examined from chicks which were either 1, 7, 14, 21 or 28 posthatch days of age. While there was a significant age-related increase in the total number of [3H]naloxone binding sites in both brain regions, the increase was greater in magnitude in forebrain than in midbrain. In both brain regions, there was a concomitant age-related decrease in the density of receptor sites without any alteration in receptor affinity for naloxone. The decrease in receptor density was more rapid in onset and was greater in magnitude in midbrain than in forebrain. PMID- 6280809 TI - Inhibition by alpha-amanitin of development of tetrodotoxin-sensitive spike induced by brain extract in cultured chick skeletal muscle cells. AB - The enhancing effect of brain extract on the development of tetrodotoxin sensitive sodium channels in cultured chick skeletel muscle cells is blocked by treatment of culture with alpha-amanitin, and RNA polymerase inhibitor. The result is interpreted as indicating that the trophic substance in the brain extract exerts its effect on the development of sodium channels through regulation of gene transcription. PMID- 6280810 TI - Differential postnatal development of mu and delta opiate receptors. AB - We found a differential postnatal development of mu and delta opiate receptors. Mu receptors labelled with low concentrations of [3H]naloxone appeared to develop earlier than did delta receptors labelled with [3H]D-Ala2-D-Leu5-enkephalin (0.5 nM). Competition binding studies also revealed a delayed appearance of delta receptors (day 12 postnatal). PMID- 6280811 TI - [Glomerular receptors and synthetic functions of the renal glomerulus]. PMID- 6280812 TI - [Hepatitis B virus and hepatoma: evidence for a cytoplasmic tumor antigen in transformed hepatocytes]. AB - A tumoural antigen (HBV/T Ag) has been found in the cytoplasm of the human hepatoma cell line PLC/PRF/5 by indirect immunofluorescence and immunoperoxidase assays using anti-sera from Senegalese patients with primary hepatocellular carcinoma. The same antigen was detected in PHC tissues. According to the data obtained with serum and tissue controls the HBV/T Ag-anti-HBV/T system seems associated with transformation of hepatocytes by hepatitis B virus. PMID- 6280813 TI - [Characterization of a yeast nuclear gene involved in the maturation of mitochondrial pre-messenger RNA of cytochrome c oxidase (author's transl)]. AB - We have analyzed a nuclear mutant of S. cerevisiae with no cytochrome c oxidase activity. The affected gene is that of a protein involved in the maturation process of the mitochondrial pre-messenger RNA of cytochrome oxidase subunit 1. PMID- 6280814 TI - [The effect of oral administration of iodized oil for prevention and treatment of endemic goiter (author's transl)]. PMID- 6280815 TI - Serological differentiation of foot-and-mouth disease virus on electron microscope grids coated with protein A and antibody. AB - A serological technique using electron microscope grids coated with protein A and antiserum was able to detect foot-and- mouth disease virus particles in oesophageal-pharyngeal fluids from infected cattle without the need for prior concentration of the sample. The technique was adapted to differentiate serologically among foot-and-mouth disease virus types A, O and C with antigen adsorbed sera. When grids were coated with heterotypic antigenadsorbed antisera, the homotypic antigen could be observed in viral specimens containing 10(5) PFU/mL, but the heterotypic antigen was not visualized until its concentration was about tenfold higher. Grids coated with the appropriate antigen-adsorbed antiserum can thus be used to indicate foot-and-mouth disease viral serotypes in specimens containing less than 10(6) PFU/mL. PMID- 6280817 TI - Identification of a strain of infectious bursal disease virus isolated from mosquitoes. AB - The Becht strain of infectious bursal disease virus was compared with a virus isolated from Aedes vexans mosquitoes and designed 743 virus. The viruses were compared with respect to cell culture host range, cellular changes resulting from viral infections, growth curves, antigenic relationship, and physicochemical characteristics. The viruses are closely comparable in all these properties, and they are considered to be strains of the same virus. In cross comparisons by the enzyme-linked immunosorbent assay, 743 virus and infectious bursal disease virus were found to be antigenically identical, confirming the results of the neutralization test. The 743 virus differs from most strains of infectious bursal disease virus in that it is nonpathogenic for chickens. PMID- 6280816 TI - Nontumoral, benign ad malignant stages of transformation of a diploid pig cell line. A review. AB - The PFT cell line originated from diploid endometrial cells and was subcultured more than 500 times. Nontumoral, benign and malignant transformation appeared at different stages. Four populations (F1, F2, F3 and F4) of cells were demonstrated sequentially by chromosomes analysis. These consisted of eight phenotypes corresponding to the sequential appearance of markers which included three chromosomal aberrations. The PFT malignant transformation in vitro is a progressive process through qualitatively different stages and may reflect neoplastic development in vivo. It is suggested that the benign and malignant transformations were under separate genetic control since they occurred after two chromosomal translocations on two different chromosomes. The spontaneous expression of a pig endogenous type-C virus which occurred concomitantly with the first translocation appeared to be related to oncogenesis. PMID- 6280818 TI - A survey of zoo mammals for antibody to rotavirus. AB - The prevalence of rotavirus antibody in zoo animals was examined. A wide range of animals were found to be seropositive, indicating a widespread susceptibility to rotavirus infection. PMID- 6280819 TI - Successful experimental challenge of dogs with canine parvovirus-2. AB - Withholding food from dogs for 24 hours prior to, and for 48 hours following oral challenge with a gut mucosal homogenate of canine parvovirus-2, was a successful means of reproducing gastroenteric signs of canine parvovirus-2 infection. Twenty one of 24 dogs, which had previously received various vaccine preparations of mink enteritis virus or were unvaccinated, and which were starved at challenge, developed soft or liquid feces with large or without large clots of mucus. Altered feces were most frequent on postexposure day 11. Seven dogs passed frank blood in their stools on one or more occasions and seven dogs vomited sporadically. Pyrexia was noted in 71.6% of the dogs on postexposure day 6 and lymphopenia was detected on postexposure day 5 or 6 in 50% of the dogs monitored. In contrast, four dogs not starved at the time of challenge remained free of gastrointestinal signs apart from one dog which passed a soft stool with scant mucus on one day, postexposure day 6. Also four dogs vaccinated with a killed canine parvovirus-2 vaccine preparation and subsequently starved at the time of challenge, remained clinically healthy. Apart from these last mentioned four dogs, all others shed canine parvovirus-2 in their feces following challenge. PMID- 6280820 TI - The failure of an inactivated mink enteritis virus vaccine in four preparations to provide protection to dogs against challenge with canine parvovirus-2. AB - Four experimental vaccine preparations comprising a strain of mink enteritis virus inactivated by either formalin or beta-propiolactone, and either adjuvanted or nonadjuvanted, failed to stimulate a consistent serum antibody response in 20 vaccinated dogs and failed to protect all but one of these dogs against oral challenge with canine parvovirus-2. PMID- 6280821 TI - Indirect hemagglutination test in equine infectious anemia. AB - An indirect hemagglutination was developed for the diagnosis of equine infectious anemia using sheep red blood cells coated with group specific virus antigen which had been highly purified by affinity chromatography. The presence of indirect hemagglutination antibodies was demonstrated in horses with equine infectious anemia since the cells were specifically agglutinated by all the serum samples obtained from experimentally infected horses. Antibodies appeared within 35 days after inoculation, and development of which coincided well with that of precipitating and complement fixing antibodies. Titer of indirect hemagglutination antibodies were ten to 320 times greater than those of precipitating antibodies. Test results could be read more clearly by the indirect hemagglutination test especially in weakly positive cases. Ninety-six samples from suspected field cases collected from every region of Japan which were positive on the immunodiffusion test were also positive on indirect hemagglutination test. Serum samples from 420 horses in one race track were examined by both the indirect hemagglutination and immunodiffusion tests to determine the reliability of the indirect hemagglutination test for diagnosis of equine infectious anemia. The same result was obtained on both tests. Based on this evidence, the indirect hemagglutination test can be employed as a very sensitive serological test for the diagnosis of equine infectious anemia. PMID- 6280822 TI - Isoelectric focusing of infectious particles of porcine pseudorabies virus strains in granulated dextran gels. AB - Isoelectric focusing of infectious particles of four strains of porcine pseudorabies viruses (Indiana, Iowa, Shope and an avirulent live-virus vaccine strains) are described. The pseudorabies virus strains exhibited great mobility in the electric field typical of viruses of the herpes group. Strains were considered electrophoretically homogeneous based on their respective isoelectric points. Maximal virus infectivity was concentrated in reproducible, stationary zones representing 73 to 86% of the total virus infectivity initially applied throughout the gels. All pseudorabies virus strains during processing and after isoelectric focusing retained their ability as whole complete particles to typically infect porcine kidney cell cultures. Virus from gel fractions produces foci in cell cultures that specifically reacted with pseudorabies virus fluorescent antibody conjugate. Prevention of foci could be demonstrated by neutralizing with pseudorabies virus monospecific antiserum. Maximal infectivity titers were demonstrated to be directly related to isoelectric points. Strain differences, in relation to virulence in swine, apparently is not related to isoelectric points. PMID- 6280824 TI - The pulmonary clearance of Pasteurella haemolytica in calves given Corynebacterium parvum and infected with parainfluenza-3 virus. AB - Four control calves were aerosolized with parainfluenza-3 and one week later with Pasteurella haemolytica. Three calves were given Corynebacterium parvum at a dose of 15 mg/m2 body surface area, infected with parainfluenza-3 virus one week later, and aerosolized with P. haemolytica two weeks after C. parvum injection. All calves were killed four hours after P. haemolytica exposure and the bacterial retention in the lung was determined. Parainfluenza-3 viral infection did not exert any suppressive effect on pulmonary clearance of P. haemolytica in six out of seven calves used. However, the bacterial colony counts in the lungs of control calves were higher (P less than 0.05) than those in calves given C. parvum. Hence, C. parvum appeared to enhance bacterial clearance. Despite the marked influx of neutrophils into the lungs after the bacterial inoculation, the neutrophil:macrophage ratio in lavage samples was less in calves given C. parvum than in the control calves. The alveolar macrophages in C. parvum treated calves were generally larger but did not differ significantly (P less than 0.05) from those in the controls. There was no significant (P less than 0.05) correlation between the percentages of alveolar macrophages and the bacterial clearance. In calves given C. parvum, bacterial clearance was enhanced in those calves which had larger macrophages. PMID- 6280823 TI - Bovid herpesvirus 2 latency: failure to recover virus from central sensory nerve ganglia. AB - Latent bovid herpesvirus 2 was sought in sensory ganglia, epithelium and lymph nodes from cattle having antibodies against bovid herpes virus 2. Tissues from eight animals were maintained in vitro as explants for 49-72 days during which all expended media was tested for virus. Three animals were pretreated with corticosteroids prior to slaughter. Infectious bovine rhinotracheitis virus was recovered from one animal, but bovid herpesvirus 2 was not detected. PMID- 6280825 TI - A "liver" antigen associated with avian erythroblastosis: binding by bentonite and precipitation with sodium dodecyl sulphate. AB - The properties of a complement fixing antigen, EbAg, extracted from erythroblastosis-affected chicken livers are described. The antigen in extracts freed of structural protein is strongly bound by bentonite, but not by barium sulphate. Strongly alkaline solutions of sodium dodecyl sulphate are required to release the antigen from bentonite. Acidification of the detergent solution precipitates the active solution precipitates the active protein. Extraction of heme from the acidified detergent precipitate by methyl-ethyl ketone further purifies the antigen. This acid detergent treatment eliminates the need to use bentonite as a purification step. PMID- 6280826 TI - The effect of diazepam on adenosine uptake and adenosine-stimulated adenylate cyclase in guinea-pig brain. AB - We have used the adenosine-stimulated adenylate cyclase of guinea-pig brain to examine the potency of diazepam as an adenosine uptake inhibitor. Diazepam at concentrations in the range 10--500 microM stimulates the production of cAMP in incubated slices of guinea-pig cerebral cortex, with maximal fivefold stimulations over basal levels by 200 microM diazepam. The increases can be largely (but not completely) blocked by the adenosine antagonist theophylline or by addition of excess adenosine deaminase to the system. It appears that the stimulation of cAMP production is due to a blockade of adenosine uptake which results in an increase in extracellular adenosine and concomitant activation of the adenosine receptor coupled to adenylate cyclase. Since the cAMP response to standard adenosine uptake blockers (dipyridamole, dilazep) can be completely blocked by theophylline or adenosine deaminase, a small component of the diazepam response cannot be explained by an adenosine effect. The concentration of diazepam at which the first significant cAMP increase occurs is 10 microM or slightly lower. This is significantly higher than the concentration of diazepam needed to saturate the pharmacologically characterized central nervous system receptors for benzodiazepines. PMID- 6280827 TI - Effects of antihypertensive clonidine congeners on alpha-adrenergic receptors. AB - Evidence in the literature suggests that the antihypertensive effects of clonidine stem from its action on alpha-2 adrenergic receptors. In order to examine this possibility we tested the effects of 13 congeners of clonidine on the binding of [3H]WB-4101 and [3H]clonidine to calf frontal cortex homogenates; [3H]WB-4101 served as a label for alpha-1 receptors while [3H]clonidine served to label alpha-2 adrenergic receptors. All the substituted imidazolines were two to three orders more potent in inhibiting the binding of [3H]clonidine than they were against [3H]WB-4101. There was a strong correlation between the antihypertensive doses of these congeners and their concentrations required to inhibit the binding of [3H]clonidine. The results are compatible with the concept that the antihypertensive action of clonidine is more likely due to interactions with alpha-2 adrenergic receptors than with alpha-1 receptors. PMID- 6280828 TI - Isolation of single atrial and ventricular cells from the human heart. AB - The single isolated heart cell has recently emerged as a model for the study of the structure and function of cardiac cells. Heart muscle cells of adult animals of various species have been successfully isolated by enzymatic digestion of intact cardiac tissue. In this paper a dissociation method that yields living cells from atrial and ventricular tissue of young and adult humans is detailed. The cells retain the morphologic features of cells in intact cardiac tissue, and they generate action potentials and contractions in response to electrical stimulation. The study of isolated human heart cells should make a valuable contribution to knowledge of the normal and diseased heart. PMID- 6280829 TI - Herpes simplex encephalitis: poor outcome despite adenine arabinoside therapy. PMID- 6280830 TI - Hypopituitarism in a patient with idiopathic hypoparathyroidism. PMID- 6280831 TI - Extensive disease small cell carcinoma of the lung: trial of non-cross resistant chemotherapy and consolidation radiotherapy. AB - Twenty-nine patients with extensive disease, small-cell carcinoma of the lung, were treated with two cycles of intensive combination chemotherapy: HexaVAC (hexamethylmelamine, vincristine, Adriamycin, cyclophosphamide). Responders received prophylactic cranial radiation (2000 rad/10 fractions) and non cross resistant chemotherapy via a schedule of alternating cycles of CMV (cyclophosphamide, methotrexate, VP-16-213) and AMV (Adriamycin, methotrexate, VP 16-213). Whenever a complete response was achieved, consolidation radiotherapy was given to the lung primary (4000 rad/20 fractions, split dose) and abdominal metastases (2000 rad/10 fractions) synchronous with CMV therapy. The complete response rate was 14% with HexaVAC, but increased to 38% during CMV/AMV. Total response rate (complete and partial) was 59% and median survival was 42 weeks. Prophylactic brain radiation prevented clinical relapse in the brain in all 14 patients who received it. However, consolidation radiotherapy failed to prevent clinical relapse in the lung and/or liver, and therapeutic brain radiation (3000 rad) failed to prevent relapse in that site. The simultaneous administration of radiotherapy and chemotherapy was well-tolerated although two patients with poor performance status died of infectious complications while leukopenic. In spite of the high response rate, durable remissions with prolonged disease free survival were rare. Further evaluation of induction, consolidation, and maintenance modes of therapy are indicated. PMID- 6280832 TI - Breast cancer skin test antigens of increased sensitivity prepared from vesicular stomatitis virus-infected tumor cells. AB - Crude membrane (CM) extracts were prepared from five cultured breast tumor lines (MDA-MB-157, MDA-MB-231, ZR75-1, HS0578T, and MCF-7) which had been infected with vesicular stomatitis virus (VSV) to augment their antigenicity. In skin test trials, CM extracts of uninfected MCF-7 cells elicited positive response in 0 of 13 (0%) tests in breast cancer patients, while VSV-MCF-7 elicited positive responses in 11 of the same 13 patients (84.6%). CM extracts of VSV-ZR75-1 and VSV-MCF-7 elicited greater delayed hypersensitivity responses (mm induration at 48 hr) in breast cancer patients than in patients with lung carcinoma or melanoma. Although the sensitivity of VSV-ZR75-1 was too low (ten of 28, or 35.7% of tests positive) to be useful as a skin test antigen, VSV-MCF-7 elicited positive responses in 30 of 38 (78.9%) tests in breast cancer patients, as compared to two of 15 (13.3%) and two of 13 (15.4%) of tests in patients with lung carcinoma and melanoma, respectively. The "virus-augmented" CM extract of cultured MCF-7 cells exhibited markedly greater sensitivity as compared to control MCF-7 extracts (P less than .005), with a high degree of specificity for breast cancer patients as compared to patients with the other neoplasms (P less than .00001). The results of skin testing with VSV-MCF-7 CM extracts demonstrated antigenic cross-reactivity with a large number of breast cancer patients, a finding of great importance for any potential immunotherapy and/or immunodiagnosis. PMID- 6280833 TI - An ultrastructural study of angiomatoid fibrous histiocytoma. AB - We studied the electron microscopy of two tumors which clinically and histologically conformed to what has been described as angiomatoid fibrous histiocytoma, a tumor thought to be of fibrohistiocytic origin. The ultrastructure of one tumor was that of a cellular angioma and the second tumor a vascular lesion with fibroblasts and histiocyte-like mononuclear cells. Our findings suggest that angiomatoid fibrous histiocytomas are basically vascular tumors with fibroblasts and other cells as secondary participants in some lesions. The behaviour of angiomatoid fibrous histiocytoma cannot be predicted from histology. Our findings of different ultrastructural components in histologically non-separable tumors lead to a question whether different ultrastructural composition is correlated with different biological behavior of these tumors. Further studies of similar cases are required if tumor morphology is to be correlated with clinical behavior. PMID- 6280834 TI - Hepatocellular carcinoma presenting as intrabile duct tumor growth: a clinicopathologic study of 24 cases. AB - We describe the features of 24 cases of hepatocellular carcinoma (HCC) with prominent intrabile duct tumor growth seen among 238 autopsy and 21 surgical cases of HCC. Progressive obstructive jaundice occurred during the course of most cases and was the presenting sign in nine. A fluctuating rise and fall of the total bilirubin was seen in two cases. The average survival time of the cases was significantly shorter than that of HCC patients without intrabile duct tumor growth (Mann-Whitney's U-test, one-tailed, P less than 0.05). The average survival time after the development of severe jaundice (total bilirubin over 10 mg/dl) was only 16 days. Intrabile duct tumor casts were located in the hepatic and/or the common bile ducts in 19 cases (79%) and in five cases were seen in the peripheral (medium to small-sized) bile ducts. Hemobilia developed in five cases (21%) and was regarded as the immediate cause of death in one. Grossly all the cases presented infiltrative or mixed (infiltrative and nodular) growth pattern. Intrabile duct tumor growth and associated marked obstructive jaundice may frequently herald the terminal phase of HCC in certain patients. In our series, approximately 40% of patients with HCC and significant jaundice had gross evidence of extensive intraductal tumor growth. In the absence of intraductal tumor growth, jaundice in HCC usually was seen in a setting of progressive terminal hepatic failure. PMID- 6280835 TI - Thorotrast-induced hepatic angiosarcoma, and combined hepatocellular and cholangiocarcinoma in a single patient. AB - A 64-year-old man developed hepatic angiosarcoma, and combined hepatocellular and cholangiocarcinoma, 36 years after Thorotrast administration. The patient presented with a large mass in the right upper quadrant. His serum alpha fetoprotein increased from 800 ng/ml to 51.2 micrograms/ml, and liver biopsy disclosed hepatocellular carcinoma. At autopsy, the liver had two different malignant neoplasms; angiosarcoma and combined hepatocellular and cholangiocarcinoma. Metastases of angiosarcoma to both lungs and of cholangiocarcinoma to periaortic lymph nodes were also seen. Interestingly, some neoplastic cells of angiosarcoma exhibited globular hyaline inclusions, which were Periodic-Acid-Schiff reaction positive and diastase-resistant. PMID- 6280836 TI - Combination chemotherapy, radiotherapy, and BCG immunotherapy in limited small cell carcinoma of the lung: a Southwest Oncology Group Study. AB - From November 1976 to March 1979 the Southwest Oncology Group treated 298 patients with limited (disease confined to the chest and encompassed by one radiotherapy port) small-cell carcinoma of the lung with combination chemotherapy and radiotherapy with or without BCG immunotherapy. Two induction chemotherapy programs were utilized: (1) cyclophosphamide, vincristine, methotrexate, fluorouracil; or (2) cyclophosphamide, doxorubicin, and vincristine. Patients received 4500 rads of radiation therapy to the bulk primary tumor and 3000 rads to whole brain followed by maintenance chemotherapy. One-half of all the patients were randomized to receive one vial (5 x 10(8)) of high viability Pasteur BCG by scarification technique given on days 8 and 15 of each 21--28 day treatment cycle. Increased granulocytopenia accompanied the addition of BCG immunotherapy. Patients receiving BCG achieved a response rate of 49% vs. those patients not receiving BCG of 44% (P = 0.579). Median response duration was 40 weeks for the BCG arms and 38 weeks for the arms without BCG; survival was no different, 42 weeks for the BCG arms vs. 50 weeks for the arms without BCG. In patients who responded to therapy and survived longer than one year, those who continued to receive BCG therapy demonstrated a slight, yet significant, survival benefit over those patients not receiving BCG (93 weeks vs. 81 weeks, P = 0.03). It appears that BCG immunotherapy has no beneficial effect on response rate, duration of response, or survival in programs using chemotherapy and radiotherapy for control of limited small-cell carcinoma of the lung except in this small group of long term survivors. PMID- 6280837 TI - Offspring of patients treated for unilateral Wilms' tumor in childhood. AB - Twenty-seven women and the wives of nine men who survived unilateral Wilms' tumor in childhood had a total of 59 live born offspring. Among the 33 infants born to women who had received orthovoltage abdominal irradiation, ten weighed less than 2500 g at birth and three died during the perinatal period. In addition, one term infant of normal weight died of complications of a breech delivery. Only one of 26 infants born to the wives of Wilms' tumor patients and unirradiated female patients weighed less than 2500 g at birth and none died. The frequency of congenital malformations and spontaneous abortions in this series was not increased, and no offspring has developed cancer. The findings suggest that the risk of Wilms' tumor is low among progeny of survivors of nonfamilial, unilateral lesions. Damage from abdominal irradiation given to girls with Wilms' tumor may predispose them to the subsequent delivery of low birthweight children. PMID- 6280838 TI - Neutralizing antibody to herpes simplex virus type 1 in patients with oral cancer. AB - Neutralizing antibody to Herpes simplex virus type 1 (HSV-1), type 2, and measles virus was measured in the serum of patients with oral cancer, patients with oral leukoplakia, and in control subjects who were smokers and nonsmokers. Significantly higher titers to HSV-1 were found in controls who smoked than in controls who did not smoke. Patients with untreated oral cancer had HSV-1 neutralizing titers similar to those of the controls who smoked, but those with later stage tumors had higher titers than those with earlier stage tumors. In patients who were tumor free after treatment for oral cancer, higher antibody titers to HSV-1 were associated with longer survival times. No association was found between clinical status and antibody to measles virus. The data are consistent with a role for both HSV-1 and smoking in the pathogenesis of oral cancer. PMID- 6280839 TI - Oat-cell carcinoma with fibrosis: evidence for release of a growth substance. AB - Recent work demonstrates that transformed cells can produce mitogenic agents capable of stimulating fibroblast proliferation. Also, work with normal cells and tissue has recently demonstrated that, among other cells, macrophages and platelets contain and elaborate potent mitogens capable of stimulating fibroblast proliferation. An unusual oat-cell carcinoma associated with large amounts of fibrous tissue was studied. Short-term cell culture of the pleural fluid was utilized to measure the production of a growth substance. Macrophages were depleted by surface adherence. A potent conditioned medium was obtained, which induced proliferation of BALB/c 3T3 cells and human diploid fibroblasts. This conditioned medium, renewed weekly, was at least as potent as Fibroblast Growth Factor 100 ng/ml every other day. It is concluded that in this human malignancy fibroblast proliferation surrounding the malignant cell is secondary to production of mitogens by the malignant cell. PMID- 6280841 TI - Treatment experience. Glioblastoma multiforme treated with 15 MeV fast neutrons. PMID- 6280840 TI - Alternating non-cross-resistant combination chemotherapy for small cell anaplastic carcinoma of the lung. AB - The development of drug resistance limits the survival or patients with small cell anaplastic carcinoma of the lung (SCLC). The present study was undertaken to overcome this problem by administering two alternating noncross resistant combination chemotherapy regimens. One-hundred-one patients were entered on study, and 98 were evaluable, with a median onstudy time of 55+ weeks. All patients received the initial combination therapy of cyclophosphamide, methotrexate, and vincristine, alternating every three weeks with Adriamycin and VP16-213 (etoposide). Radiation therapy was not a standard part of protocol. Thirty-two patients had regional disease (LD), and 66 had extensive disease (ED). Overall, 76% of patients responded to this therapy with 30 (31%) complete remission (CR) and 44 (45%) partial remissions (PR); the respective CR and PR rates were 31% and 50% for LD, and 30% and 42% for ED patients. Myelosuppression was the principal toxicity with a leukocyte nadir of 2.0 X 10(9)/1 in 10% of cycles. Septicemia in six neutropenic ED patients with progressive disease contributed to the only treatment-related deaths. Patients entering CR had a median survival greater than 51 weeks (range, 8-150+); 58+ for LD and 49+ for ED patients. Patients in PR had respective median survivals of 33+ (overall), 43+ (LD), and 24+ (ED) weeks. Forty patients have had relapses with initial sites being local sites in 35%, and neurologic in 38%. Although this protocol has not discernibly delayed the onset of drug resistance, the problem should be considered when new protocols are designed in SCLC. PMID- 6280842 TI - Solitary dural plasmacytoma with conspicuous cytoplasmic inclusions. AB - A solitary intracranial plasmacytoma originated from the left parietal meninges of a 67-year-old woman; the patient had a four-month history of generalized seizures. Immunoelectrophoresis, performed nine months after surgery, revealed a monoclonal IgG band in the cerebrospinal fluid (CSF) but not in the blood. A peculiar feature of the tumor cells was the presence of abundant hyaline, eosinophilic cytoplasmic inclusions. Ultrastructurally, these inclusions consisted of electron-dense floccular material within distended cisterns of endoplasmic reticulum (ER). These ultrastructural features resemble "thesaurocytes" originally described by Paraskevas et al. in IgA myeloma. Tumors composed mainly of such inclusion-bearing cells are extremely rare. Only a fraction of the tumor cells stained positively for IgG with the immunoperoxidase technique; most inclusion-bearing cells did not stain. It is therefore unlikely that the electron-dense material is stored immunoglobulins. PMID- 6280843 TI - Contiguous malignant astrocytoma and Wilms'-like tumor in the brain. AB - A case of primary brain tumor composed of two contiguous neoplasms in presented. At operation, a nodular tumor was embedded in the infiltrating tumor within the brain parenchyma. With light and electron microscopy, the nodular tumor was similar to Wilms' tumor (nephroblastoma). The infiltrating tumor was malignant astrocytoma. Autopsy revealed, besides the recurrence of malignant astrocytoma in the brain and its subarachnoid dissemination, extracranial metastases to the abdominal cavity, liver, lung, and bone marrow. Recurrent and metastatic tumors were glioblastoma multiforme, which was more malignant than the surgical specimen. The possibility of metastatic Wilms' tumor from the kidney was completely ruled out by extensive autopsy survey. The authors present an extremely rare tumor including detailed observations on the electron microscopic appearance of the tumor; the histogenesis of these tumors is briefly discussed. PMID- 6280844 TI - Alpha-1-antitrypsin deficiency in Southern African hepatocellular carcinoma patients. An immunoperoxidase and histochemical study. AB - An increased frequency of alpha-1-antitrypsin deficiency (AATD) has been reported in patients with hepatocellular carcinoma (HCC) in Scandinavia and England. Using a specific immunoperoxidase technique for alpha-1-antitrypsin (AAT) and periodic acid-Schiff (PAS) with and without diastase predigestion, the authors examined nonneoplastic autopsy liver tissue from 58 black southern African patients with HCC and from 54 controls. No periportal PAS-positive diastase-resistant globules containing AAT (specific for AATD) were found in liver tissue from either group. A finely granular diffuse cytoplasmic AAT staining (not removed by diastase predigestion) was present in hepatocytes in a "checkerboard" pattern within the lobule in 33 (57%) HCC patients and in 20 (37%) controls (P greater than 0.1), and was particularly prominent adjacent to tumor in 11 HCC patients. Alpha-1 antitrypsin was present in neoplastic cells in 18 of the 40 HCC examined (45%). These findings suggest no major role for AATD in the etiology of HCC in southern Africa. PMID- 6280845 TI - Chemotherapy of advanced ovarian carcinoma: initial experience using a platinum based combination. AB - Thirty-five previously untreated patients with FIGO Stage III or IV ovarian carcinoma were treated with cis-diamminedichloroplatinum (II), Adriamycin and cyclophosphamide (PACe). Following four or five cycles of PACe, patients were assessed clinically for response. Twenty-one (60%) of the patients attained a clinical complete remission (CCR), seven (20%) a partial remission, and seven (20%) did not respond. The median survival of all patients was 19 months, 6.4 months for partial and nonresponders, and 26.5 months for clinical complete responders. Six patients remain relapse-free for more than two years. Patients entering into the study during the second year underwent a second-look procedure if they attained a CCR. Ten patients in CCR were assessed for second look; two refused, and eight underwent a laparotomy. Three of eight were in complete remission after second look, and a fourth patient who had microscopic disease in one ovary was rendered free of disease by surgery. Subjective toxicity was marked due to severe nausea and vomiting. Renal toxicity was not troublesome and myelosuppression was moderate. Ototoxicity and peripheral neuropathies were seen in seven patients. PACe is a highly effective induction regime but randomized studies are required to show that improved survival will result from it's use. PMID- 6280846 TI - S-100 protein in granular cell tumors (granular cell myoblastomas). AB - Outside the central nervous system S-100 is found only in Schwann cells and satellite cells of ganglia. It has also been demonstrated in Schwannomas and neurofibromas but is absent from soft tissue tumors of non-neural origin. S-100 protein was looked for in granular cell myoblastomas using an immunohistochemical technique in an attempt to further elucidate the histogenesis of these tumors. All tumor cells in the ten tumors studied were intensely stained with antiserum to S-100 including one with some malignant features. These results support the idea that granular cell myoblastomas arise from Schwann cells. PMID- 6280847 TI - Histologic typing of lung cancer in Louisiana. AB - To determine whether histologic patterns differed in the high- and low-lung cancer mortality parishes (counties) of Louisiana and whether the findings in the state differed from those in other parts of the United states, we studied the available histopathologic materials for 272 persons of the 815 who died of lung cancer in ten southern, nonurban Louisiana parishes during a seven-year period from 1971-1977. Squamous-cell carcinoma and small-cell anaplastic carcinoma were the most common tumor types, closely followed in frequency by adenocarcinoma, confirming reports by other investigators of a change during the past decade in the prevalence of various histopathologic types of lung cancer. The distribution of histopathologic types was not different for high- and low-mortality parishes but differed significantly from other areas of the U. S. Three persons had diagnoses consistent with pleural mesothelioma. Occupational histories obtained from relatives showed that one of those persons was a homemaker and the other two were sugarcane farmers with no discernable exposure to asbestos. PMID- 6280848 TI - Tumor-induced hypoglycemia: a result of ectopic insulin production. AB - A 73-year-old woman with a pelvic tumor of nerve cell origin who presented with recurrent episodes of hypoglycemia was demonstrated to have inappropriately elevated plasma insulin and hypoglycemia. The tumor contained Insulin (RIA), proinsulin and secretory type of granules. Removal of the tumor promptly produced hyperglycemia. Also no islet cell tumor was found in the pancreas, suggesting the tumor as the site of ectopic insulin production. PMID- 6280849 TI - Cardiac tamponade due to malignant pericardial effusion in breast cancer: a case report. AB - Cardiac tamponade resulting from malignant pericardial effusion is an uncommon initial presentation of various extracardiac malignancies. Only twenty-nine cases of extracellular malignancies with this initial presentation have been previously published. Lung carcinoma leads as the most common malignancy involved, followed by carcinoma of the stomach, pancreas, kidney and ovary, mediastinal rhabdomyosarcoma, malignant lymphoma and leukemia. This report describes a case of breast carcinoma in a 47-year-old woman who initially presented with cardiac tamponade. To the best of the authors' knowledge, no similar case has even been reported in the literature. PMID- 6280850 TI - Effect of age at first childbirth on risk of developing specific histologic subtype of breast cancer. AB - Epidemiologic variables related to breast cancer risk were assessed in a case control study of 332 women with breast carcinoma and 1353 comparison women. Risk factors for breast cancer as a whole included nulliparity, late age at first childbirth, early age at menarche, late age at menopause, personal history of benign breast disease, family history of breast cancer, and among postmenopausal women, body weight. These risk factors were then analyzed with respect to histologic subtype of breast cancer involved, i.e., duct-derived or lobular tumors, to determine whether the association between any of the risk factors and breast cancer varied according to histopathologic subtype. Histologic subtype for the 316 cases reviewed included 284 duct cancers and 32 lobular carcinomas. Although slight differences were noted among some of the risk factors and the variety of cancer, none of the differences was marked except for the variable age at birth birth. For ductal carcinoma, the risk was highest among nulliparous women and decreased the younger a woman was at the time she gave birth to her first child. The risk of infiltrating lobular carcinoma, however, was lowest among nulliparous females or those who had given birth at a young age and increased the older a woman was when she gave birth to her first child. PMID- 6280851 TI - Cancer in Liberia: a review of cases registered from the Liberia Cancer Registry 1973-1977. PMID- 6280852 TI - Activity of bis-carbamoyloxymethyl derivatives of pyrroles and pyrrolizines against human tumor xenografts in nude mice. AB - The activities of the pyrrolizines, Compounds I and II, and the pyrroles, Compounds IV and V, are reported in nude mouse-grown HT-29 human adenocarcinoma of the colon, DO 1 human oat carcinoma of the lung, and CL 1 human duct cell carcinoma of the breast (coded as CX-1, LX-1, and MX-1, respectively, by the National Cancer Institute). Compounds II and IV were active against all three experimental tumors; both compounds produced significant tumor regression in the human breast tumor xenograft and Compound IV caused tumor regression in the human lung tumor xenograft. PMID- 6280853 TI - Biological behavior of MM1 hamster melanoma. AB - We have reported evidence recently for a high-affinity receptor for glucocorticoid Malignant Melanoma No. 1 hamster melanoma and suggested that tumor growth was facilitated by adrenal steroids. This report characterizes the behavior of Malignant Melanoma No. 1 following manipulation of the pituitary adrenal axis in vivo. Bilateral adrenalectomy significantly retarded tumor growth. Hypophysectomy also significantly reduced tumor growth. Silastic implants of hydrocortisone in intact hamsters produced a dose (7 to 28 micrograms/day) related increase in tumor growth. Implants releasing a low dose (3 micrograms/day) of dexamethasone also increased tumor growth. Chronic exposure of adrenalectomized and intact hamsters to a high dose (125 micrograms/day) of desoxycorticosterone acetate also produced a significant increase over adrenalectomized and sham-adrenalectomized controls. In contrast, chronic administration of adrenocorticotropic hormone and alpha-melanocyte-stimulating hormone to intact hamsters did not significantly alter melanoma growth. These observations support the suggestion that adrenocorticosteroids influence the growth of Malignant Melanoma No. 1 hamster melanoma and provide a model for studying the regulation of growth of a glucocorticoid-positive neoplasm originating outside the reticuloendothelial system. PMID- 6280854 TI - Characterization of transformed cells and tumors by proton nuclear magnetic resonance spectroscopy. AB - Cultured acute lymphoblastic leukemic cells give a well-resolved proton nuclear magnetic resonance spectrum characteristic of isolated plasma membranes. We demonstrate that the signals, in the spectrum of whole cells, arise predominantly from the plasma membrane and that cells transformed by pokeweed mitogen have membranes which are significantly less rigid than are normal human peripheral blood lymphocytes. Normal thymus, malignant thymus, and a leukemic T-cell line have been compared by proton nuclear magnetic resonance spin echo experiments, and the normal thymus was found to differ. Cells transformed by the Epstein-Barr virus can also be characterized and shown to differ from the leukemically transformed cells by spin echo experiments. Since no probe molecule was required to obtain these results, this is the first definitive evidence that the structure and fluidity of the plasma membranes change as a result of transformation of lymphocytes. Proton nuclear magnetic resonance spectroscopy can now be used to compare the effect of different mitogens on T- and B-lymphocytes as well as to monitor the effects of drugs, metals, etc., on the plasma membrane of transformed lymphocytes. PMID- 6280855 TI - Enhanced tumor growth in vivo by a factor in human platelets and rat liver. AB - Liver cell supernatant and platelet extract can promote the in vivo growth of herpes simplex virus type 2-transformed hamster embryo fibroblast cells. Delineation of necessary growth factors for tumor cells in vivo may open up new strategies to inhibit malignant cell growth. PMID- 6280856 TI - Murine leukemia virus-mediated transformation of a mouse epithelial cell line MMC E. AB - Mouse embryo epithelial cells, MMC-E, were malignantly transformed in culture with MuLV derived from C3H/MCA 5 cells chemically induced by 5-iododeoxyuridine. The frequency of transformation was significantly enhanced if infection was followed by treatment with 100 ng of 12-O-tetradecanoylphorbol-13-acetate per ml. The virus-transformed cells retained their high plating efficiency in soft agar in the absence of 12-O-tetradecanoylphorbol-13-acetate and formed neoplasms in nude mice. PMID- 6280857 TI - Identification of a fish protein associated with a kinase activity and related to the Rous sarcoma virus transforming protein. PMID- 6280858 TI - New human papilloma virus isolated from epidermodysplasia verruciformis lesions. AB - Human papilloma virus (HPV) was isolated from red plaques of a patient (N. F.) with epidermodysplasia verruciformis. Electron microscopic examination showed characteristic particles of papilloma virus as icosahedrons about 45 nm in diameter. DNA was extracted from these particles, and closed-circular DNA (Form I) was purified by centrifugation in CsCl containing ethidium bromide. The molecular weight of the DNA was about 5.0 x 10(6). A physical map of the HPV DNA was constructed using several restriction enzymes. The restriction endonuclease cleavage pattern of the HPV DNA was different from those of other types of HPV reported thus far, suggesting that the isolate was a new, as yet unclassified, HPV. PMID- 6280859 TI - HLA antigens in patients with germ cell cancers of the testis. AB - The expression of HLA-A, -B, -C, and -DR antigens has been analyzed in 145 unrelated Caucasian patients with germ cell tumors of the testis. Eighteen of these patients had pure seminoma, while the remaining patients had nonseminomatous tumors with embryonal carcinoma, teratocarcinoma, choriocarcinoma, and/or yolk sac components, with or without seminoma. Increases were noted in the frequencies of Aw33, B5, DR5, and DRw6 among the patients with pure seminoma, A3 and B7 among the patients with embryonal carcinoma with or without seminoma, and Aw32 among the patients with yolk sac tumor components. A decrease in the frequency of HLA-DR3 was noted in all patients subgroups, although none of these differences were statistically significant after correction for the number of antigens tested. HLA typing results for three affected brothers of patients indicate that, in each family, the affected sibling pair share at least one HLA haplotype. The etiological and prognostic significance of this finding and of the increases in a few HLA antigen frequencies in particular patient groups and the overall decreases in DR3 remain to be determined. PMID- 6280860 TI - Human central nervous system distribution of cis-diamminedichloroplatinum and use as a radiosensitizer in malignant brain tumors. PMID- 6280861 TI - Treatment of small-cell carcinoma of the lung monitored by sequential flow cytometric DNA analysis. PMID- 6280862 TI - Characteristics of Hodgkin's disease-derived cell lines. AB - In the last 3 years we were able to establish five long-term in vitro cell cultures from biopsy specimens taken preterminally from four patients with histologically proven Hodgkin's disease (nodular sclerosing type, clinical stage IVB). Four of the lines are continuously proliferating in vitro; one culture stopped growth for unknown reasons after 7 months. When culture conditions were modulated, the first culture, L 428, gave rise to two sublines: L 428 KS, after adaptation to calf serum, and L 428 KSA, permanently growing as an adherent monolayer line after treatment with a phorbol ester (12-O-tetradecanoylphorbol-13 acetate) for 3 weeks. Cell-marker analysis by conventional means (SIg, cIg, rosette formation, Epstein-barr virus reactivity, cytochemistry, phagocytosis, and lysozyme production) and with monoclonal antibodies directed against various human lymphoid, myeloid, and monocytoid antigens showed that the tested cell lines are clearly different from all hitherto described hematopoietic lines; they most likely represent a cell type resembling an early myeloid-monocytoid progenitor cell. Conditioned medium of the L 428 cells and its two sublines showed colony-stimulating factor activity and suppression of spontaneous cell mediated cytolysis of L 428 KS and K 562 cells. PMID- 6280863 TI - Developmental increase of digitalis receptors in guinea pig heart. AB - The apparent differences of digitalis tolerance in human newborns or young animals and adults may be due to the difference in either, the sensitivity of Na+,K+ -ATPase to digitalis, the binding characteristics of the Na+,K+ -ATPase receptor sites or the number of receptor sites itself. Sarcolemmal Na+,K+ -ATPase enriched membrane preparations from guinea pig hearts of various age groups were investigated for Na+,K+ -ATPase activity, the ability of ouabain to inhibit the enzyme and the 3H-ouabain binding characteristics of the receptor sites. There was a continuous developmental increase in Na+,K+ -ATPase activity in fetal (45 days fetus, 1.2 mumol Pi . mg-1 protein . h-1) and newborn (2.4 mumol Pi . mg-1 protein . h-1) reaching maturity at the age of 21 to 25 days (9.1 mumol Pi . mg-1 protein . h-1). The ability of digitalis to inhibit the enzyme, however, was same throughout the development with 50% inhibition occurring at 0.1 mumol . litre-1 concentration of ouabain. The sodium stimulated 3H-ouabain binding was saturable and exhibited a single class of receptor sites. The number of digitalis receptor sites increased progressively in fetal (45 days fetus, 2.2 pmol . mg-1 protein) and newborn (9.1 pmol . mg-1 protein) reaching mature levels at the age of 13 to 25 days. The affinity of the Na+,K+ -ATPase receptor sites for digitalis (Kd, 8.68 - 9.33 x 10(-9) mol . litre-1) remain unchanged during development. The data obtained is consistent with the concept that Na+,K+ -ATPase is the receptor site for digitalis and suggests that factors other than the developmental increase in Na+,K+ -ATPase may be relevant to explain the age-related differences in digitalis sensitivity. PMID- 6280865 TI - Thickness of enamel layers removed by HClO4 etching. PMID- 6280864 TI - Effect of topical application of urea peroxide on caries incidence and plaque accumulation in rats. PMID- 6280866 TI - Calmodulin and the adenylate cyclase-phosphodiesterase system. PMID- 6280867 TI - Plant and fungal calmodulin: Ca2+-dependent regulation of plant NAD kinase. AB - Although little is known about the role(s) of second messengers, including free Ca2+, in plant cells there has been increasing evidence for a role for Ca2+ in metabolic regulation in plants. The recent demonstration that the Ca2+-binding protein, calmodulin exists in extracts of higher plants and basidiomycete fungi provides a basis for understanding Ca2+-dependent metabolic regulation in plant cells. In this review we summarize the similarities and differences of plant, fungal and mammalian calmodulin. We also discuss the known in vitro functions of calmodulin in higher plants. A Ca2+-calmodulin-dependent NAD kinase has been purified to homogeneity from extracts of pea seedlings and shown to be absolutely dependent upon calmodulin and microM levels of free Ca2+ for activity. The available evidence suggest that this Ca2+-calmodulin-dependent NAD kinase is the major form of plant NAD kinase and that this regulatory enzyme is localized in the chloroplast. A model is presented which predicts that the rate of photosynthesis is regulated by a receptor-mediated change in the level of chloroplastic free Ca2+ upon illumination. Free Ca2+, acting as a second messenger, forms a Ca2+-calmodulin complex thus converting calmodulin to its active conformation. This Ca2+-calmodulin complex then activates chloroplastic NAD kinase resulting in an increased NADP/NAD ratio. PMID- 6280868 TI - SINEs and LINEs: highly repeated short and long interspersed sequences in mammalian genomes. PMID- 6280869 TI - Poliovirus-induced inhibition of host-cell protein synthesis. PMID- 6280870 TI - Phosphorylation of vinculin by pp60src: what might it mean? PMID- 6280871 TI - Clusters of genes encoding mouse transplantation antigens. AB - We constructed a cosmid library from BALB/c mouse sperm DNA and isolated 64 cosmid clones with cDNA probes for transplantation antigens (class I molecules). Of these clones, 54 mapped into 13 gene clusters containing 36 distinct class I genes and encompassing 837 kilobases of DNA. One gene cluster mapped to the L region and a second cluster with seven genes to the Qa-2,3 region of the major histocompatibility complex. Restriction map and Southern blot analyses suggest that there are subgroups of class I genes. Using a 5' flanking sequence of the L gene as a hybridization probe, we show the L gene to be present in mouse strains expressing this antigen but deleted or mutated in strains failing to express it. Our data suggest that gene duplication and deletion presumably by homologous but unequal crossing-over has altered the size and organization of the class I clusters in different mouse strains and probably is an important mechanism for generating polymorphism in these genes. Analysis of the 36 class I genes with cDNA probes specific for the 5' and 3' ends shows that the exon encoding the third external domain is far more conserved than those encoding the first and second external domains of the transplantation antigen. These differences in variability have interesting functional implications. PMID- 6280872 TI - Multiple sequences related to a constant-region kappa light chain gene in the rabbit genome. AB - Four allelic genes control kappa light chain allotypes in the rabbit. Amino acid sequence studies have revealed an extensive divergence (22%--33%) in the alternative forms of the kappa constant region (b4, b5, b6 and b9). Furthermore, independent studies have shown that a rabbit could express a wrong allotype. To assess the hypothesis that b allotypes are encoded by duplicated genes with a polymorphic control mechanism, we have analyzed the DNAs of four different homozygous rabbits using the Southern blot hybridization technique, with a cloned b4 C kappa probe. DNAs of individual b4, b5, b6 and b9 rabbits were cleaved with Eco RI, Kpn I and Pst I restriction endonucleases. Comparative analysis of restriction patterns shows that all four rabbit DNAs contain multiple DNA fragments hybridizing with the probe under low- and high-stringency washing conditions. In addition, each restriction pattern is distinct, suggesting that the C kappa genes are organized differently in animals expressing different allotypes. PMID- 6280873 TI - The uvrB gene of Escherichia coli has both lexA-repressed and lexA-independent promoters. AB - We have found that the uvrB gene of Escherichia coli is transcribed from at least two promoters, which we call P1 and P2. Transcription from P1 begins with an ATP at +1, and transcription from P2 begins primarily with a GTP at -31. A binding site for the lexA protein (LEXA), located between the -35 sequence and Pribnow box of P2, regulates transcription from this promoter. In vitro, LEXA inhibits transcription from P2 but has no detectable effect on transcription from P1. A third promoter, P3, was also detected at -341; transcription from P3 is toward uvrB but terminates in vitro in the region of the LEXA binding site. The binding of LEXA to P2 inhibits transcription from the P3 promoter even though several hundred nucleotides separate the two promoters. The data suggest that a transcribing RNA polymerase stalls when it reaches the repressor-operator complex but remains bound to the DNA, causing a jamming of RNA polymerases between P3 and the repressor-operator complex at P2. The physiological significance of P3 is unknown. PMID- 6280874 TI - Partially deficient methylation of cytosine in DNA at CCATGG sites stimulates genetic recombination of bacteriophage lambda. AB - Lambda bacteriophages grown on arl mutants of Escherichia coli ("Arl-" phages) display intermediate levels of cytosine methylation: less 5-methylcytosine (m5C) than phages grown on wild-type bacteria ("Arl+" phages) but more than phages grown on dcm mutants, and thus lacking the methylated sequences (Cm5CATGG) characteristic of E. coli K-12 bacteria ("Dcm-" phages). "Arl-" phages are one twelfth as resistant to Eco RII restriction (recognition site CCATGG) as "Arl+" phages, but 40-fold more resistant than "Dcm-" phages. Chromatographic analyses show the 5-methylcytosine content of "Arl-" DNA to be one third that of "Arl+" DNA. Altered cytosine methylation frequency correlates with two previously described properties of "Arl-" phages, increased genetic recombination and unusual sensitivity of phage DNA to endonuclease S1, which are absent in phages grown on dcm or dcm arl bacteria. Methylated/unmethylated heteroduplex DNA prepared in vitro (one strand from Eco RII-modified phages/one from "Dcm-" phages) is highly recombinogenic but not S1-sensitive. We hypothesize that hemimethylated CCATGG sites in "Arl-" DNA are necessary and sufficient for enhanced recombination, and necessary but not sufficient for S1 sensitivity. PMID- 6280875 TI - DNA sequence required for efficient transcription termination in yeast. AB - The cyc1-512 mutation is a 38 base pair deletion in the 3' nontranslated region of the CYC1 locus in the yeast Saccharomyces cerevisiae. The deletion occurred between two 7 bp directly repeated sequences. The cyc1-512 mutant produces approximately 10% of the normal amount of the CYC1 gene product, iso-1-cytochrome c, and produces 5%--10% of the normal steady-state amount of CYC1 mRNA. Most of the mRNAs in cyc1-512 are longer at their 3' ends by up to 1000 nucleotides, suggesting that the 38 bp deletion in cyc1-512 prevents proper transcription termination. The improper transcription termination is shown to cause converging transcription between CYC1 and an adjacent gene. The fact that all of the aberrantly sized mRNAs in cyc1-512 are polyadenylated leads us to suggest that polyadenylation may be coupled to transcription termination in yeast. We have uncovered a consensus sequence between the region deleted in cyc1-512 and the 3' nontranslated regions of some but not all yeast genes, and discuss the possible role of this sequence in transcription termination. PMID- 6280876 TI - Transcription and accurate polyadenylation in vitro of RNA from the major late adenovirus 2 transcription unit. AB - Nucleoprotein complexes with in vitro transcription activity were isolated from HeLa cells late in lytic infection with Adenovirus type 2 (Ad2). Both polymerase II and polymerase III were active in these extracts, and greater than 85% of the labeled RNA was Ad2-specific. Electrophoretic analyses and Southern blot analyses demonstrated that RNA complementary to the entire 30 kb late transcription unit including RNA near the presumed termination site was synthesized. The addition of DRB (5,6,dichloro-1-beta-D-ribofuranosyl-benzimidazole) in vivo prior to the isolation of the complexes resulted in accumulation of polymerase II at the promoter proximal sites, but nascent chains started in vivo in DRB were successfully elongated in vitro. Approximately 10% of the RNA labeled in vitro contained poly(A), and the length of poly(A) was very similar to that of nuclear RNA isolated from Ad2-infected cells. The in vitro sites of poly(A) addition were specific--labeled poly(A)-terminated RNA molecules ended at a point on the genome coincident with previously mapped poly(A) sites of mRNAs produced in vivo. In addition, the polyadenylation enzyme (or enzymes) cosediment with the nucleoprotein complexes during sucrose gradient centrifugation since gradient purified complexes synthesize poly(A) containing RNA in vitro in the absence of any added nuclear extract. PMID- 6280877 TI - Abnormally spliced messenger RNA in erythroid cells from patients with beta+ thalassemia and monkey cells expressing a cloned beta+-thalassemic gene. AB - The reduced beta-globin synthesis characterizing the beta+ thalassemia phenotype has been shown to be caused by anomalous processing within the small intervening sequence (IVS1) of the beta-globin mRNA precursor. The beta-globin gene from such patients contains a single base substitution within IVS1, located 22 bp from the 3' junction between IVS1 and exon 2, creating an alternative splice site within IVS1 and resulting in retention of the 3'-terminal 19 bases of IVS1. We have identified this abnormally spliced mRNA in the reticulocyte RNA of two patients with beta+ thalassemia, by S1 nuclease mapping and primer-extension analysis. Moreover, a cloned beta+-thalassemic gene preferentially generated the anomalously spliced RNA when expressed in monkey kidney cells. The anomalously spliced RNA constituted approximately 80%--90%, and normal beta RNA approximately 10%--20%, of the total beta mRNA. In contrast, the small amount of beta mRNA present in reticulocytes from such patients consisted predominantly of normal beta mRNA. These results suggest that the reduced amount of normally functioning beta mRNA present in such patients results from preferential processing at the alternative splice site, with subsequent instability, reduced nuclear processing and/or inadequate cytoplasmic transport of the abnormal RNA species. PMID- 6280878 TI - Allele-specific, selective amplification of a ribosomal RNA gene in Tetrahymena thermophila. AB - The amplification of ribosomal DNA during development of the somatic macronucleus in Tetrahymena thermophila was analyzed by genetic and molecular biological techniques. We have identified an alternate form of the rDNA, structurally distinguishable from the wild-type by an extra cutting site for Bam HI in its nontranscribed spacer. The altered rDNA was inherited in crosses in a simple Mendelian fashion, consistent with the presence of only one rRNA gene copy per haploid genome in the micronucleus. We therefore define a locus for the rRNA structural gene, the rdnA locus, with the allele determining the alternate form designated rdnA1. In over 95% of T. thermophila clones heterozygous for the rdnA locus in the micronucleus (rdnA1/rdn+), the macronucleus, which develops from a division product of this micronucleus, contained almost exclusively rdnA1-type amplified palindromic rDNA molecules. The rdnA1 allele is thus almost always dominant over the rdn+ allele with respect to amplification. This genetic variant of the rdnA locus was used to show that the single, free, nonpalindromic rRNA genes, which are synthesized during rDNA amplification, are derived from micronuclear gene copies from both chromosomal homologs. We therefore conclude that in these heterozygotes, selective amplification of the rdnA1 allele is not caused by the production of only one type of free, single rRNA gene during amplification. PMID- 6280879 TI - Chicken oviduct progesterone receptor: location of specific regions of high affinity binding in cloned DNA fragments of hormone-responsive genes. AB - We have used a DNA-cellulose competitive binding assay to measure the extent of displacement of the chicken oviduct progesterone-receptor complex from calf thymus DNA-cellulose by purified cloned fragments of genomic DNA. Several DNA fragments from hormonally responsive genes coding for egg-white proteins were found to be efficient competitors for either crude or partially purified receptor complexes when compared with calf thymus DNA. Data obtained with deletion mutations constructed in vitro allowed delineation of a specific region necessary for strong competition, located 250--300 bp upstream from the mRNA startsite of the ovalbumin gene. Sequence homologies with this 5'-upstream region were observed in other fragments of the ovalbumin, conalbumin, ovomucoid, X and Y genes, which were also efficient competitors. Based on a comparison of such sequences of homology, a consensus sequence that may constitute a region binding progesterone-receptor complex has been constructed: ATCCCTTATTATTCTGGTTTGTA. The results suggest that specific double-stranded DNA sequences are recognized by the oviduct progesterone-receptor complex in vitro, and are relevant to the question of whether specific DNA sequences are directly involved as genomic binding sites for steroid receptors. PMID- 6280880 TI - Influence of sex hormones on Coxsackie B-3 virus infection in Balb/c mice. PMID- 6280881 TI - Immunoglobulin properties of Epstein-Barr virus transformed human umbilical cord and adult peripheral blood lymphocytes. PMID- 6280882 TI - Macrophage-mediated cytolysis of erythrocytes in the guinea pig. II. Role of oxidative burst products in macrophage cytotoxicity activated by lectins and phorbol ester derivatives. PMID- 6280883 TI - Effect of DDT on adenylate cyclase activity in rat liver cell membranes. PMID- 6280884 TI - Chemical interactions with herpes simplex type 2 virus: enhancement of transformation by hydrazine and 1,2-dimethylhydrazine. AB - Quantitative assays for the morphological transformation of 3T3 Swiss mouse cells by herpes simplex type 2 virus (HSV-2) were employed to examine the effect on cell transformation of chemical carcinogens and suspected carcinogens. Exposure of the cells to the chemical compound, followed by virus infection, resulted in enhancement of transformation when compared to that observed with chemical or virus alone. Enhancement occurred in tests utilizing either UV light-inactivated HSV-2 (strain 333) or a temperature-sensitive (ts) mutant of HSV-2 (A8(293)]. A series of seven ts-mutants were tested and exhibited varying degrees of transformation. Enhancement of transformation occurred in cells treated with hydrazine (HZ) and 1,2-dimethylhydrazine (SDMH). No enhancement occurred when cells were treated with monomethylhydrazine, 1,1-dimethylhydrazine and the jet fuels JP-5, JP-10, RJ-4 and RJ-5. A strong time dependence after treatment was demonstrated with some enhancement seen at 6 h after chemical treatment but the greatest enhancement appeared when virus infection began after 24 h of chemical exposure. PMID- 6280885 TI - Inhibition of cyclic AMP phosphodiesterase by lignans. PMID- 6280886 TI - Disposition of methylglyoxal bis(guanylhydrazone) (MGBG, NSC-32946) in man. AB - A high-pressure liquid chromatographic method was utilized to determine the concentration of the antileukemic agent methylgloxal bis(guanylhydrazone) (MGBG, NSC-32946) in tissue samples obtained at autopsy from patients who received MGBG. In a patient with cholangiocarcinoma who received one course of MGBG (500 mg/m2), the highest drug concentration was found in normal liver tissue. However, the drug concentration in intrahepatic tumor tissue was only 10% of that found in uninvolved liver. In a patient with acute myelogenous leukemia (AML) who received 12 courses of MGBG therapy, highly infiltrated lymph node tissue was found to contain the highest concentration of MGBG. High concentrations of the drug were also found in liver, spleen, kidney, adrenal, and thyroid. The drug penetrated well into normal brain tissue. After repeated administration, high drug concentrations were found in cerebral and cerebellar gray matter. These studies suggest that there is no selective uptake of MGBG into solid tumors early after drug administration and provide a pharmacologic rationale for testing this agent against endocrine and intracerebral tumors in man. PMID- 6280887 TI - Compartmentalization of the adenosine pool of dog and rat hearts. PMID- 6280888 TI - Relationship between beta-adrenergic receptor numbers and physiological responses during experimental canine myocardial ischemia. AB - In the present study, we evaluated the physiological responsiveness of the increased numbers of beta-adrenergic receptors in ischemic canine myocardium to in vivo stimulation by (-)-isoproterenol and epinephrine. After 1 hour of temporary proximal left anterior descending coronary artery occlusion and during a 15-minute period of reflow, dogs received (1)-isoproterenol intravenously at a rate sufficient to increase their heart rates 20--40 beats/min. Following the infusion of isoproterenol, myocardial tissue was obtained from the LV ischemic and nonischemic regions for measurement of beta-adrenergic receptor numbers, cyclic AMP content, and phosphorylase b to a conversion. beta-Adrenergic receptor numbers were significantly increased in the left ventricular (LV) ischemic tissue. The administration of (-)-isoproterenol was associated with significant increases in cyclic adenosine monophosphate content and phosphorylase b to a conversion in the LV ischemic tissue. Also, the administration of (-)-epinephrine significantly increased the phosphorylase b to a conversion in ischemic tissue over the nonischemic tissue and this conversion was blocked by pretreatment with (+/-)-propranolol. These data suggest that, in this experimental model, the increased numbers of beta-adrenergic receptors in canine LV ischemic tissue are capable of translating physiological responses when they are activated with an appropriate agonist in vivo. PMID- 6280889 TI - An experimental model for congestive heart failure after encephalomyocarditis virus myocarditis in mice. AB - Severe myocarditis was induced in inbred BALB/c mice inoculated with the M variant of encephalomyocarditis (EMC) virus. The mortality rate was maximal on the fourth day, then decreased gradually, but increased again between the eleventh and fourteenth days. Gross myocardial lesions were seen on the surface of the ventricles in 62 of 125 mice (49.6%) after the fifth day. These myocardial lesions were observed more frequently in the dead mice (46 of 49, 93.9%). Cavity dimensions and wall thickness were measured in two groups of mice with myocarditis. On days 5-7, the cavity dimensions of the right (RV) and left (LV) ventricles in inoculated mice (0.92 +/- 0.51 mm and 1.21 +/- 0.18 mm, respectively) were significantly larger than those in controls (RV 0.54 +/- 0.17, LV 1.01 +/- 0.15; p less than 0.05). The wall thickness of the RV (0.46 +/- 0.09, controls 0.64 +/- 0.11; p less than 0.001) and the LV (0.97 +/- 0.13, controls 1.12 +/- 0.19; p less than 0.05) was significantly decreased. On days 8-14, dilatation of the LV was more pronounced (1.48 +/- 0.37, p less than 0.005) than during days 5-7, and the interventricular septum was also thinner. Pleural effusion, ascites and congestion of the lungs and liver were noted, and death seemed due to congestive heart failure. This study is the first documentation of congestive heart failure after viral infection is an experimental animal. PMID- 6280890 TI - Hemodynamic and radionuclide effects of acute captopril therapy for heart failure: changes in left and right ventricular volumes and function at rest and during exercise. AB - Although the resting hemodynamic effects of captopril in congestive heart failure are known, little information is available about the hemodynamic response to captopril during exercise or about changes in noninvasive measurements of the size and function of both ventricles. In this study, 14 stable New York Heart Association class III patients were given 25 mg or oral captopril. Rest and exercise hemodynamic measurements and blood pool scintigrams were performed simultaneously before and 90 minutes after captopril. The radionuclide studies were analyzed for left and right ventricular end-diastolic volumes, end-systolic volumes, ejection fractions and pulmonary blood volume. The primary beneficial responses at rest were decreases in left and right ventricular end-diastolic volumes from 388 +/- 81 to 350 +/- 77 ml (p less than 0.01) and from 52 +/- 26 to 43 +/- 20 volume units (p less than 0.01), respectively, and in their corresponding filling pressures, from 24 +/- 10 to 17 +/- 9 mm Hg and 10 +/- 5 to 6 +/- 5 mm Hg (both p less than 0.001). Although stroke volume did not increase significantly, both left and right ventricular ejection fractions increased slightly, from 19 +/- 6% to 22 +/- 5% and from 25 +/- 9% to 29 +/- 11%, respectively (both p less than 0.01). During exercise, similar changes were noted in both hemodynamic and radionuclide indexes. Thus, in patients with moderate symptomatic limitation from chronic heart failure, captopril predominantly reduces ventricular volume and filling pressure, with a less significant effect on cardiac output. These effects persist during exercise, when systemic vascular resistance is already very low. Radionuclide techniques are valuable in assessing the drug effect in these subjects, particularly when ventricular volumes are also measured. PMID- 6280891 TI - The relationship between the strength of the human heart beat and the interval between beats. AB - In 15 patients undergoing cardiac catheterization and pacing tests, the left ventricular (LV) pressure and its maximum rate of rise (LV dP/dt max) were measured with catheter-tip manometers. Atrial or ventricular pacing at a single steady frequency (the priming frequency) was followed by a test pulse at a varying interval (test pulse interval). In 14 subjects in whom it was examined, the contractile response after the test pulse increased with test pulse interval to reach a maximum plateau value--the optimum contractile response (OCR). In five cases, further prolongation of the test pulse interval decreased the contractile response. The optimum test pulse interval occurred at 800-900 msec. An increase in the priming frequency before the introduction of the test pulse caused a progressive increase in OCR, in contrast to the minor effects on LV dP/dt max of the control beats. Similar results were recorded in four other patients in whom contractile response was assessed from the rate of rise of right ventricular pressure. These results indicate that with tachycardia, the interval between beats is insufficient to allow maximum contractile performance (presumed to be activated by calcium ions) to develop. The true effect of increasing heart rate is only revealed by the relationship between OCR and the preceding frequency of contraction. PMID- 6280892 TI - Comparison of cardiovascular response to combined static-dynamic effort, postprandial dynamic effort and dynamic effort alone in patients with chronic ischemic heart disease. AB - The cardiovascular responses to combined static-dynamic effort, postprandial dynamic effort and dynamic effort alone were evaluated by upright bicycle ergometry during equilibrium-gated blood pool scintigraphy in 24 men, mean age 59 +/- 8 years, with chronic ischemic heart disease. Combined static-dynamic effort and the postprandial state elicited a peak cardiovascular response similar to that of dynamic effort alone; work load 643 +/- 156 and 638 +/- 161 vs 650 +/- 153 kg-m/min, respectively; heart rate 147 +/- 14 and 145 +/- 14 vs 143 +/- 17 beats/min; systolic pressure 195 +/- 26 and 200 +/- 25 vs 197 +/- 25 mm Hg; and rate-pressure product 286 +/- 48 and 292 +/- 55 vs 282 +/- 52. Heart rate, intraarterial systolic and diastolic pressures, rate-pressure product and ejection fraction were similar for the three test conditions at the onset of ischemia and at peak effort. The prevalence and extent of exercise-induced ischemic left ventricular dysfunction, ST-segment depression, angina pectoris and ventricular ectopic activity were also similar during the three test conditions. Direct and indirect measurements of systolic and diastolic blood pressure were highly correlated. The onset of ischemic ST-segment depression and angina pectoris correlated as strongly with heart rate alone as with the rate-pressure product during all three test conditions. The cardiovascular response to combined static-dynamic effort and to postprandial dynamic effort becomes more similar to that of dynamic effort alone as dynamic effort reaches a symptom limit. If significant ischemic and arrhythmic abnormalities are absent during symptom limited dynamic exercise testing, they are unlikely to appear during combined static-dynamic or postprandial dynamic effort. This simplifies, the task of formulating guidelines for physical effort in patients with chronic ischemic heart disease, especially in providing "clearance" to perform avocational and vocational tasks involving combined static-dynamic and postprandial dynamic effort. PMID- 6280893 TI - 5'-Nucleotidase activity enhanced by manganese and magnesium ions with inosine monophosphate substrate. PMID- 6280894 TI - Human proteins sensitive to neoplastic transformation in cultured epithelial and fibroblast cells. AB - We resolved the [35S]methionine polypeptides synthesized by normal [lung fibroblasts (WI-38), amnion cells] and transformed [SV40 transformed WI-38, AMA (spontaneously transformed amnion cells)] cultured human cells, using two dimensional gel electrophoresis under conditions in which about 1300 polypeptides could be reproducibly separated. These studies demonstrated important changes in the relative proportions of several polypeptides that are present both in normal and transformed cells. Of a total of 400 common polypeptides that we quantitated for each cell type, 53 (22 basic and 31 acidic) varied by 40% of more in both cell pairs. Among these, we have identified vimentin (IEF 26), cyclin (IEF 49), and a tropomyosin-related polypeptide (IEF 52). No new major polypeptide was detected in the transformed cells, at least at the level of resolution currently achieved by this technique. Similar but qualitative studies of [32P]orthophosphate-labeled proteins revealed that, of 250 analyzed, only seven common phosphoproteins, including phosphovimentin (IEF 26e), varied consistently in both cell pairs. These results strengthen our previous conclusion that transformation results in changes in the relative proportions of polypeptides synthesized in normal and transformed cells rather than in the appearance of new polypeptides in transformed cells. PMID- 6280895 TI - Automated assay for alpha 1-antitrypsin with N-alpha-benzoyl-DL-arginine-p nitroanilide as trypsin substrate and standardized with p-nitrophenyl-p' guanidinobenzoate as titrant for trypsin active sites. AB - alpha 1-Antitrypsin is the most abundant of several serum protease inhibitors. Its deficiency is associated with an increased incidence of emphysema in adults, jaundice in newborns, and childhood cirrhosis. We describe an automated functional assay for the Instrumentation Laboratory's Multistat III Microcentrifugal Analyzer with N-alpha-benzoyl-DL-arginine-p-nitroanilide as trypsin substrate. The assay is standardized in terms of moles of trypsin active sites inhibited per liter of serum, by use of a chromogenic titrant for trypsin active sites, p-nitrophenyl-p'-guanidinobenzoate. The method is rapid, precise, and independent of trypsin supplier, and results correlate well with those by a manual chromogenic and a nephelometric assay. PMID- 6280896 TI - Dexamethasone suppression of adrenocortical function. AB - To investigate the potency of dexamethasone in suppressing adrenocortical function, we gave various evening doses to laboratory staff members. Plasma corticotropin, cortisol, and dehydroepiandrosterone sulfate concentrations were measured the following day, to quantitate the degree of suppression after both short- and long-term treatments. We found that the minimum amount for effective suppression was 1 mg for a single evening dose, 0.5 mg per day for a long-term evening dose. On these regimens plasma corticotropin and cortisol were consistently suppressed, but plasma dehydroepiandrosterone sulfate was suppressed only with long-term treatment. PMID- 6280897 TI - Improved micromethod for assay of serum angiotensin converting enzyme. AB - We describe conditions for determining angiotensin converting enzyme (EC 3.4.15.1) in serum, by liquid chromatography. Serum (10 microL) is assayed with the artificial substrate hippuryl-glycyl-glycine (30 mmol/L) in a 50 mmol/L "HEPES" buffer solution with a high salt content (300 mmol of NaCl and 400 mmol of Na2SO4 per liter), at pH 8.0. The resulting enzymic activity is ninefold that of the currently popular spectrophotometric assay of Cushman and Cheung as modified by Lieberman (Am. J. Med. 59: 365-372, 1975). The hippuric acid end product is separated from the substrate by reversed-phase liquid chromatography and measured spectrophotometrically at 228 nm. o-Methyl hippuric acid is used as internal standard. The mean value for 100 normal control subjects was 317 (SD 96) nmol of hippuric acid released per milliliter of serum per minute. The enzyme activity is greater in newborns (p less than 0.05) and has a tendency to decrease with age. This partly automated method, which is optimized with regard to activity and detection, can be used in clinical routine. PMID- 6280898 TI - The steroid response to controlled adrenal stimulation in congenital adrenal hyperplasia. AB - Adrenal steroidogenesis has been studied in vivo in eleven patients aged 13-68 years with 21-hydroxylase deficiency, in one patient with 11 beta-hydroxylase deficiency and in ten female control subjects. Serum levels of the delta 5 3 beta hydroxysteroids, pregnenolone (Pe), 17 alpha-hydroxypregnenolone (17Pe), dehydroepiandrosterone (DHEA) and androstenediol (Adiol) and their delta 4 3-keto counterparts, progesterone (Po), 17 alpha-hydroxyprogesterone (17Po) androstenedione (Adione) and testosterone as well as of 11-deoxycortisol and cortisol were measured during acute adrenal suppression with dexamethasone followed by stimulation with synthetic 1-24 ACTH. In the seven patients with 21 hydroxylase deficiency who were on adequate glucocorticoid therapy, grossly exaggerated responses of 17Po and Po to ACTH were nevertheless preserved. In contrast, there was a grossly subnormal response of 17Pe, DHEA and Adiol to ACTH, and low basal levels of DHEA-sulphate. In the untreated patients the response of 17Pe and DHEA was normal. The Adione response was exaggerated in untreated and normal in treated cases. Similar findings obtained in the patient with 11 beta hydroxylase deficiency who was studied after 6 weeks without replacement therapy. Our findings demonstrate that production of adrenal steroids that are associated with the adrenarche is not exaggerated in untreated CAH, and is grossly suppressed in treated cases. These findings are compatible with the hypothesis that intra-adrenal cortisol may initiate and/or maintain production of the delta 5 steroids by the zona reticularis that occurs in the human adrenarche. PMID- 6280899 TI - Guanfacine and other centrally acting drugs in antihypertensive therapy; pharmacological and clinical aspects. AB - Centrally acting antihypertensive agents exert an agonist action on alpha adrenoceptors in certain areas of the brain, thereby reducing sympathetic outflow and lowering blood pressure without paralysing peripheral homeostatic control mechanisms. Some also stimulate peripheral alpha-adrenoceptors, both postsynaptic and presynaptic. Guanfacine, a representative member of this class of drugs, resembles clonidine in most of its basic pharmacological properties. In some respects, however, clear differences exist and may account for an improved therapeutic usefulness. In cats the two drugs have a different site of action within the CNS. Unlike clonidine, guanfacine does not inhibit dopamine turnover in the corpus striatum of the rat and its hypotensive effect is not inhibited by central H2-receptor blockade. In rat EEG studies guanfacine is much less sedative than clonidine. The newer drug shows a higher selectivity for (peripheral) presynaptic alpha-adrenoceptors than clonidine. On a weight basis guanfacine is about 10 times less potent in lowering blood pressure, but at equipotent doses its antihypertensive effect lasts longer. With an elimination half-life of approximately 18 to 21 h in man guanfacine is suitable for once-a-day treatment. The long duration of action is likely to explain the lack of rebound hypertension in chronically treated spontaneously hypertensive rats as well as the very low incidence and the remarkably mild nature of withdrawal symptomatology in man. PMID- 6280901 TI - Sodium and human hypertension. PMID- 6280900 TI - Effects of the new oral angiotensin converting enzyme inhibitor MK-421 in human hypertension. AB - "MK-421" a new oral converting enzyme inhibitor was administered to 16 hypertensive patients in doses ranging between 2.5-40 mg once daily for periods of 6-18 weeks. The blood pressure, plasma renin activity, plasma angiotensin II, aldosterone and angiotensin converting enzyme activity were assessed before and during treatment. Blood pressure normalized in 11 patients and decreased significantly in the remaining 5, without significant changes in heart rate or orthostatic hypotension. At 24 hrs after the first effective dose, plasma renin activity remained elevated to 228% whereas plasma angiotensin II was suppressed to 58%, aldosterone was suppressed to 40% and angiotensin converting enzyme activity was suppressed to 16% of the baseline. Magnitude of blood pressure decrement achieved with the maximal dose did not correlate with baseline plasma renin activity levels. No side-effects were noted during the 6-18 week period of observation. PMID- 6280902 TI - Angiotensin-I converting enzyme activity in the sera of captopril-treated hypertensive patients. AB - The blood pressure and serum angiotensin-I converting enzyme (SACE) activity were measured in captopril-treated hypertensive patients at frequent intervals. Inhibition of SACE was observed in patients responding to the treatment with lowering of blood pressure as well as in nonresponding cases. It was therefore concluded that a mechanism not depending on the formation of angiotensin-II by ACE is responsible for the high blood pressure persisting in the captopril treated nonresponding hypertensive patients. SACE activity in sera of captopril treated patients recovers from inhibition when stored in frozen state at -20 degrees c. An apparent dissociation was therefore observed when regeneration of SACE activity during storage of sera was not taken into account. Meaningful activities are therefore obtained only if the assay is performed without prolonged storage. PMID- 6280903 TI - (Na+,K+)-activated adenosinetriphosphatase and hypertension in DAHL salt- sensitive and -resistant rats. AB - (Na+,K+)-ATPase activity was compared in Dahl salt-sensitive (S) and salt resistant (R) rats. When S and R rats were maintained on 1% NaCl diet their blood pressures at 5 weeks of age were similar and their renal microsomal (Na+,K+) ATPase activities were also similar. At 6 months of age, on 1% NaCl diet, S rats have markedly elevated blood pressure compared to R and renal microsomal (Na+,K+) ATPase activity was suppressed in S compared to R. Feeding 8% NaCl diet for 5 weeks induced hypertension in young S rats but failed to alter renal or brain (Na+,K+)-ATPase activity. Heart (Na+,K+)- ATPase activity was elevated in S compared to R rats regardless of salt intake of blood pressure. It appears unlikely that mutations in the structural locus for the renal (Na+,K+)-ATPase molecule are involved in the strain specific differences in susceptibility to salt-induced hypertension since the physical-chemical properties of the enzyme from the two strains were found to be similar. Since renal (Na+,K+)-ATPase activities were unchanged by salt feeding and resultant blood pressure changes in young S rats, the suppressed renal (Na+,K+)-ATPase activity seen only in old S rats is probably a response to prolonged renal damage and not a response to "natriuretic factors." Elevated heart (Na+,K+)-ATPase in S-rat hearts is unexplained. PMID- 6280906 TI - [Primary hyperparathyroidism: clinical and laboratory aspects (author's transl)]. PMID- 6280904 TI - Altered inotropic responsiveness to isoproterenol of hearts from spontaneously hypertensive rats during development. AB - The characteristics of the inotropic responses to isoproterenol (Iso) were investigated in isolated hearts from embryonic normotensive (NT) and spontaneously hypertensive (SH) Wistar-Kyoto rats at 14, 16, 18 and 20 days of gestation and from rats 3 and 10 weeks after birth and in ventricular strips prepared from hearts of newborn and 1-week-old rats. With rates of all preparations maintained constant by electrical stimulation, concentration and age dependent increases in contractility in response to Iso, capable of being blocked by propranolol, were observed. In embryonic hearts, irrespective of Iso concentration or embryonic age, similar effects were observed in hearts from NT and SH animals. However, in all postnatal preparations, the effects of Iso on the SH strain were quantitatively different from those on hearts of NT rats of the same age. As early as 12 hr after birth and through 10 weeks of age, most concentrations of Iso induced inotropic effects that were significantly greater in hearts of the SH than the NT strain and the slopes of the linear portions of the concentration-response curves were significantly steeper. These findings indicate that functional beta adrenergic inotropic receptors are present in the embryonic hearts of both strains and undergo changes during development that are strain dependent. PMID- 6280907 TI - [Biological activities of vitamin D3 metabolites and their new analogs (author's transl)]. PMID- 6280908 TI - [Vitamin D3 metabolite receptor in breast cancer (author's transl)]. PMID- 6280909 TI - [A case of congenital adrenal hypoplasia (author's transl)]. PMID- 6280910 TI - [Antibodies of the TSH receptor in Graves' disease (author's transl)]. PMID- 6280911 TI - [Serum human calcitonin level in patients with non-thyroidal cancer (author's transl)]. PMID- 6280912 TI - Systemic treatment of tinea versicolor with ketoconazole in forty-four patients. PMID- 6280913 TI - Immunoglobulin synthesis in nude (nu/nu), nu/+ and reconstituted nu/nu mice infected with a demyelinating strain of Semliki Forest virus. AB - Infections with the avirulent (A7/74) strain of Semliki Forest virus which causes primary demyelination of the central nervous system in mice have been studied further in nude athymic (nu/nu) mice and their immunocompetent (nu/+) litter mates to measure the production of immunoglobulins. This has been done by radial immunodiffusion and enzyme-linked immunosorbent assays. Half the nude mice examined were able to synthesize specific IgG but at levels 1,000-fold lower than their nu/+ littermates. The majority of nude mice reconstituted with spleen cells from nu/+ mice 1 day before infection with virus were able to synthesize specific IgG nearly as well as the nu/+ animals. PMID- 6280914 TI - Hemolytic uremic syndrome in two postmenopausal women taking a conjugated estrogen preparation. PMID- 6280915 TI - Value of pre-treatment and follow-up skeletal scintigraphy in operable breast cancer. PMID- 6280916 TI - Tc-99m pertechnetate uptake in a thymoma: case report. AB - A case is reported to Tc-99m pertechnetate accumulation within an anterior mediastinal thymoma during a search for substernal goiter. This reemphasizes the non-specificity of Tc-99m pertechnetate uptake and the need for caution in using this agent to detect ectopic thyroid tissue. PMID- 6280917 TI - Clinical utility of a multigated modified anterior projection in the detection of left ventricular inferior and apical wall motion abnormalities. AB - Recent evidence indicates that the left anterior oblique projection (LAO) multigated radionuclide ventriculogram (RVG) underestimates presence and extent of apical and inferior left ventricular (LV) wall motion abnormalities. We investigated, prospectively, the sensitivity and specificity of a modified anterior projection (MAP), which incorporates cephalad tilting. Thirty-three consecutive patients undergoing cardiac catheterization suspected to have coronary artery disease were studied with RVG, using both the MAP and LAO views. LAO views were analyzed using the ejection fraction image (REFI), and the regional ejection fraction (REF) of the inferoapical region. The MAP studies were analyzed using stroke volume image (SVI) to evaluate apical and inferior LV regions. Results were as follows: (Formula: see text), Both intraobserver and interobserver variabilities were comparable to those of conventional angiographic studies used in detection of apical and inferior asynergy. It is concluded that the multigated MAP offers additional information about abnormalities of the LV inferior and apical regions. PMID- 6280918 TI - Comparison of effects of marihuana cigarettes to three different potencies. AB - Marihuana cigarettes containing 1.32%, 1.97%, and 2.54% delta 9 tetrahydrocannabinol (THC) were smoked by six experienced marihuana users at weekly intervals in a double-blind cross-over design under laboratory conditions. Puff duration, number of puffs taken, duration of inhalation holding, interval between puffs, and duration of smoking were recorded for each cigarette smoked. The portion of each cigarette remaining after smoking was weighed and analyzed to determine THC content. Subjective ratings of the "high" achieved and the heart rate acceleration induced by smoking the marihuana were measured. The plasma concentrations of THC and of its principle metabolite, 11-nor-delta 9-THC-9 carboxylic acid (9-carboxy THC), were determined by radioimmunoassay of blood samples drawn at frequent intervals for 6 hr. The results indicate that, irrespective of the potency of the marihuana, the pattern of smoking was much the same. The magnitude of the subjective high, heart rate acceleration, THC, and 9 carboxy THC plasma concentrations were proportional to potency. This dose response was particularly clear between the 1.32% and the 2.54% cigarettes. Peak plasma concentrations of THC consistently occurred 7 to 8 min after initiation of smoking and declined thereafter despite continued smoking for another 6 to 10 min. Peak subjective high and peak heart rate acceleration occurred several minutes after the end of smoking and at a considerable interval after maximal THC plasma concentrations were reached. PMID- 6280919 TI - Ranitidine kinetics in normal subjects. AB - A 1-mg/kg IV bolus injection and a 150-mg (one tablet) oral dose of ranitidine were given to seven normal subjects. At least 1 wk separated the two doses. Ranitidine disappeared from plasma with a half-life of about 2.5 hr. Half of the oral dose was effectively absorbed and half of the absorbed amount was found unchanged in urine. Total body clearance was 10.1 ml/min/kg. Urinary clearance was the same after oral and intravenous doses (6.4 and 6.9 ml/min/kg, P greater than 0.10). Intravenous ranitidine kinetics included three phases, with a central distribution volume of 0.2 l/kg and a total distribution volume of 1.2 l/kg. Absorption kinetics were apparently order zero: of the 150-mg dose, 75 mg was absorbed during 5 hr at a constant rate of 15 mg/hr. PMID- 6280921 TI - Decrease in the tubular reabsorption of calcium with evidence for compensatory hyperparathyroidism in X-linked hypophosphataemia in mice. PMID- 6280920 TI - Erythrocyte sodium transport in chronic renal failure. PMID- 6280922 TI - CT demonstration of scirrhous carcinoma of stomach: a case report. PMID- 6280923 TI - Rh hemolytic disease-Connecticut, United States, 1970-1979. Centers for Disease Control. PMID- 6280924 TI - Catechol estrogens and thrombosis: lack of a direct effect of 2-hydroxyestradiol on platelets. AB - Women who use the oral contraceptive pill are at risk for a variety of thromboembolic complications, but the precise mechanism is unclear. Since 2 hydroxyestradiol is a major metabolite of estradiol and possesses some of the structural and functional properties of catecholamines, we examined whether this compound might influence thrombogenesis, in particular whether it would exert an epinephrine-like effect on the platelet. In vitro, 2-hydroxyestradiol does not cause or potentiate aggregation or thromboxane release, nor does it prevent the rise in cyclic AMP caused by anti-aggregatory prostaglandins. While 2 hydroxyestradiol does not exert a direct effect on the platelet in vitro, it may play a role in thrombosis in women who take oral contraceptives through another mechanism. PMID- 6280925 TI - Severe bronchiolitis probably caused by varicella-zoster virus. AB - An unusual, severe pneumonia probably caused by varicella-zoster virus is reported in a 19-year-old previously healthy man. The diagnosis was based on high titer of varicella-zoster antibodies in serum, and demonstration of varicella zoster antigen from lung biopsy specimen. The uncommon feature in the pathophysiological course of the disease was the selective hypercarbia that responded well to bronchodilator therapy with theophylline. Furthermore, the patient had no skin manifestations during his illness. PMID- 6280926 TI - Ultrasonographic evaluation of the kidney. AB - The kidney is a readily accessible organ to the ultrasonographer. In the past several years, improvements in instrumentation have allowed detailed visualization of the normal renal architecture as well as disease processes involving the kidney. Ultrasonographic evaluation should be considered a complementary modality to the intravenous pyelogram, and in cases where contrast material is contraindicated, it may become a primary imaging technique of the kidneys. The indications for a renal ultrasonic examination, techniques, and methods of imaging the kidney as well as normal anatomy will be presented. A review of several renal abnormalities will include mass lesions, hydroenphrosis, pediatric applications, assessment of transplants, and interventional techniques. An integrated radiologic approach to renal disease, including intravenous pyelography, ultrasonography, computed tomography, and angiography, will be suggested. PMID- 6280928 TI - Genetic analysis of tumorigenesis. VIII. Suppression of SV40 transformation in cell hybrids and cytoplasmic transferants. AB - Intraspecies somatic cell hybrids of BALB/c mouse 3T3 and SV40-transformed embryonic fibroblast (SVT2) cells were analyzed for transformation-associated properties and their tumorigenic potential in nude mice. In confirmation of our earlier findings, hybrids expressing the viral T-antigen were not suppressed for the ability to clone in medium with 1% serum. In contrast, division rate in medium with 1% or 10% serum, anchorage independence, cytochalasin-sensitive growth control, and tumorigenicity were suppressed noncoordinately, and the extent of suppression varied from one hybrid to another. Suppression was not simply determined by the increased chromosome content of the hybrid cells, nor was suppression correlated with rearrangements of the integrated viral sequence (SAGER et al., 1981a, b). Similar results were found in cytoplasmic transferants expressing T-antigen. Four independent transferants and subclones derived from them varied in the extent of suppression of anchorage independence and tumorigenicity. In both hybrids and transferants, a low serum requirement for clonal growth apparently was determined solely by expression of SV40 T-antigen, but other transformation properties, as well as tumorigenicity, appeared to require multiple changes in the cellular genome for their expression. These changes must occur during or after viral integration, since they are not expressed in uninfected 3T3 cells. PMID- 6280927 TI - Kainic acid: The neurotoxic breakthrough. PMID- 6280929 TI - Frequency of abnormalities of cortisol secretion and water metabolism in patients with small cell carcinoma of the lung and other malignancies. AB - Previous studies have suggested that ectopic production of adrenocorticotropic hormone (ACTH) or antidiuretic hormone (ADH) may occur commonly in patients with small cell carcinoma of the lung (SCCL) and that evidence of such production may be elicited only by provocative tests of water excretion and adrenal function. We studied 28 patients with SCCL and 29 patients with other cancers. Adrenal function, assessed by measuring the 8 am plasma cortisol, the 8 am to 4 pm diurnal variation in plasma cortisol, and the suppressibility of the 8 am plasma cortisol following administration of 1 mg of dexamethasone, was found to be abnormal in 28.5, 71, and 25 percent, respectively, of the patients with SCCL, compared with 18, 65, and 29.5 percent in patients with other types of cancer (P greater than 0.3). The possibility of ectopic ADH secretion was assessed by a standard water loading test, which showed excretion impairment in 60 percent of patients with SCCL and 68 percent of patients with other cancers (P greater than 0.9). Neither the stage of neoplastic disease, sites of metastatic deposits, nor performance status of the patients correlated with abnormalities of water and cortisol metabolism, indicating that such abnormalities are common in patients with all types of cancer. These data do not suggest that subclinical disturbances of adrenal function or water excretion are characteristic of any histologic type of cancer. The precise mechanism(s) underlying these abnormalities are unknown. PMID- 6280930 TI - Adenovirus bronchiolitis in Manitoba: epidemiologic, clinical, and radiologic features. AB - We reviewed our experience with 41 children hospitalized from 1974 to 1978 for adenovirus (ADV) bronchiolitis. Thirty-two patients (78 percent) were native Indians between four and 12 months old. In 18 of the 41 patients (43.9 percent) acute complications developed. The five fatal cases (12.2 percent) were confined to native children. The initial chest roentgenograms showed lobar consolidation in 35 patients (85.4 percent). Atelectasis developed in five (12.2 percent) during hospitalization. Sixteen of 25 patients (64 percent) with adequate radiologic follow-up examination had subsequent pneumonias or showed residual chronic changes. The reasons for the predilection of ADV bronchiolitis in native Indian children and the precise effect on subsequent airway function in survivors are unknown and require further study. We emphasize the importance of ADV as a cause of bronchiolitis in native Indian children. Furthermore, this report focuses attention on the contribution of this disease to the spectrum of chronic pulmonary disorders in the pediatric group. PMID- 6280931 TI - Pharmacokinetics of a new cephalosporin, cefotaxime (HR 756) in patients with different renal functions. PMID- 6280932 TI - [Hemorrhagic shock]. PMID- 6280933 TI - [Is "blind left-pancreatic resection" for insulinoma still indicated?]. AB - One of the main problems encountered in the surgical treatment of insulinoma is that of locating them within the pancreas, since about 10% of these tumors are occult. Modern pre- and intraoperative methods of identifying of hormone producing pancreatic tumors remove the need for 'blind resection'. Complete exploration of the pancreas, biopsy, intraoperative toluidin-blue-0 staining, glucose monitoring, and especially selective pancreatic venous blood sampling with insulin radioimmunoassay make it possible to locate nearly all tumors. Percutaneous transhepatic portal venography and selective portal blood sample collection for insulin analyses should be done routinely before planning a reintervention when organic hyperinsulinism persists. PMID- 6280935 TI - [The change of plasma cAMP and cGMP in chronic aplastic anemia (author's transl)]. PMID- 6280934 TI - Localization in mouse-L-Cell chromosomal sites of transferred immunoglobulin genes. AB - Mouse thymidine kinase negative (tk-) L-cells were cotransformed with two different kappa light chain genes (cloned from mouse myeloma) and the tk gene from Herpes simplex virus I. (Transformation is defined as change in the genotype by introduction of foreign DNA.) About 80% of the tk+ -transformants contained varying amounts of transferred kappa light chain sequences, one transformant about 150 copies per genome. The transferred immunoglobulin genes appear to be organized in a nucleosomal substructure, as deduced from digestion experiments with micrococcal nuclease. In situ hybridization experiments revealed, that the transferred genes are not distributed randomly across the chromosomes of the recipient cell. Instead they are clustered at one or a few chromosomal locations. PMID- 6280936 TI - [Rapid immune electron microscopic detection for viral particles of viral hepatitis A and B (author's transl)]. PMID- 6280937 TI - Problems in the congenital lactic acidoses. AB - The congenital lactic acidosis form a heterogeneous group of inborn errors that includes defects of gluconeogenesis, the pyruvate dehydrogenase complex, the Krebs cycle and the respiratory chain. These disorders are not easily classified because of the absence of specific metabolites, difficulties in providing suitable tissue specimens and technical problems with the enzyme assays. The commonest causes of lactic acidosis due to inborn errors are the deficiencies of glucose-6-phosphatase and fructose bisphosphatase, which present with hypoglycaemia, lactic acidosis and hepatomegaly. Pyruvate carboxylase and phosphoenolpyruvate deficiencies vary considerably in both clinical expression and biochemical findings. Neurological symptoms predominate in defects of the pyruvate dehydrogenase complex, and some cases of the spinocerebellar ataxias may be due to partial defects of the pyruvate and 2-oxoglutarate dehydrogenase complexes. PMID- 6280938 TI - [Reaction of pulmonary macrophages activated by BCG to silica: I. Research in quantitative changes of intracellular acid phosphatase (author's transl)]. PMID- 6280939 TI - Cell cycle and 5'-Nucleotide phosphodiesterase activities in rat liver. AB - The postnatal developments of liver and serum 5'-nucleotide phosphodiesterase (5' NPD) in Fischer 344 rats were studied. The liver enzyme activities were found to correlate well with the growth of the liver as measured by the wet weight and percentages of liver cells in the G2/M phases of the cell cycle as determined by flow cytofluorometry. Therefore, this enzyme may be intimately related to the growth of the liver. Although the specific activities of the serum enzyme are three orders of magnitude less than that of the liver enzyme, very good correlation was obtained between the total serum and liver enzyme activities. This finding, therefore, suggest that the main origin of the serum enzyme is from liver. Multiple isozymes were detected in both liver and serum 5'-nucleotide phosphodiesterase after polyacrylamide gel electrophoresis. PMID- 6280940 TI - Pathological secretion of the breast. PMID- 6280941 TI - [Establishment and some characteristics of a hepatoma cell line (QGY-7703) (author's transl)]. PMID- 6280942 TI - [Pathological studies on early primary hepatocellular carcinoma. An analysis of 37 resected small lesions (author's transl)]. PMID- 6280943 TI - [Small oat cell lung cancer and long term survival with regard to TNM staging--a report of 11 cases (author's transl)]. PMID- 6280944 TI - Placental and fetal pathology in infectious mononucleosis. A possible indicator for Epstein-Barr virus teratogenicity. AB - Placentae and membranes were studied microscopically in five cases of pregnancy interruption due to maternal infectious mononucleosis in the first 2 months of pregnancy. This viral disease, caused by Epstein-Barr virus and apparently rare in pregnancy, induced specific placental lesions. In the decidua, these lesions were manifested by perivasculitis and necrotizing deciduitis. The membranes exhibited slight to moderate chorionitis. In the villi, endovasculitis, perivasculitis, and occasional vascular obliteration were found, as well as mononuclear and plasma cell villitis, with many large atypical vacuolated cells resembling plasma cells. Three fetuses were studied, two of which exhibited myocarditis. The fact that there were placental lesions in all cases studied, and that two of the fetuses exhibited myocarditis, must stimulate further research concerning the possible teratogenicity of this virus in man. PMID- 6280945 TI - Adenosarcoma of the ovary. A light- and electron-microscopic study with review of the literature. AB - This is a light- and electron-microscopic study of a large ovarian adenosarcoma in a 50-year-old woman as well as a review of the literature. The tumor showed a variety of mullerian type epithelia and a spectrum of stromal patterns including nonspecific fibroblastic, and endometrial type stroma. The latter ranged from normal-appearing to low-grade sarcoma. This study emphasizes the relationship of ovarian adenosarcoma to cystadenofibroma, on the one hand, and to malignant mixed mullerian (mesodermal) tumor, on the other. It appears that the mitotic count which is used as the primary means for the classification of smooth muscle and stromal tumors of the uterus, has less value in the assessment and prognostication of ovarian adenosarcomas. PMID- 6280946 TI - [Use of nucleoside di- and monophosphates as substrates in RNA synthesis by replicative complexes of the encephalomyocarditis virus]. PMID- 6280947 TI - [Properties of the receptive structure controlling sex hormone activity and sexual selectivity of biologically active substances]. PMID- 6280948 TI - [Ketoconazole in the treatment of pulmonary aspergillosis and aspergilloma: mycological observations (author's transl)]. PMID- 6280949 TI - [Meralgia paraesthetica in degenerative arthritis of the hip-joint (author's transl)]. PMID- 6280950 TI - [On the occurrence of the viral diarrhea/mucosal disease virus in cervines in Rheinland-Pfalz (author's transl)]. PMID- 6280951 TI - [A contribution to the egg drop syndrome of laying chickens. 1: Clinical and serological findings (author's transl)]. PMID- 6280952 TI - [A contribution to the egg drop syndrome of laying chickens. 2. Macroscopic and histological findings (author's transl)]. PMID- 6280953 TI - [Causal relationship and pathogenesis of leukosis, kidney tumors, ovarian neoplasms and osteopetrosis following infection with avian leukosis viruses]. PMID- 6280954 TI - [Investigations on the stress syndrome at turkeys--influence of ACTH and excitement upon glucose concentration and white blood cells (author's transl)]. PMID- 6280955 TI - [Herpesvirus infections in animals]. PMID- 6280956 TI - [Abortion, stillbirth, runting, deaths of lambs in two sheep herds--border disease in the Federal Republic? (author's transl)]. PMID- 6280957 TI - [Investigations on inactivated zincbacitracin (author's transl)]. PMID- 6280958 TI - [Possibilities and limits of MD/VD and IBR vaccination in a cattle fattening stable. Observations of a case in practice (author's transl)]. PMID- 6280960 TI - [Alcohol and the peripheral nerves]. PMID- 6280959 TI - [Effects of alcohol on the nerve cell membrane]. PMID- 6280961 TI - Neuromuscular junction dysfunction in primary hyperparathyroidism: a new hypothesis. PMID- 6280962 TI - Intracellular localization of prolactin receptor and prolactin in the rat ovary by immunocytochemistry. AB - The localization of the PRL receptor as well as of PRL has been studied by immunoperoxidase techniques in the ovaries of cycling, pregnant, and lactating rats. Specific antisera to the receptor and to the hormone were used. By light microscopy, immunostaining for the PRL receptor coincided with that for the hormone. Staining was found intracellularly in most components of the ovary, except the theca, and was most striking in the luteal cells. Both PRL and its receptor were concentrated heavily in the ovum. Beginning 24-36 h postpartum, there was a change in the pattern of luteal cell staining, with a shift in the intensity of staining products to the periphery of the luteal cell, giving a ring appearance to these cells. The results suggest roles for PRL in ovarian function involving both maintenance of the corpus luteum and maturation of the ovum. This study also demonstrates the intracellular localization of a polypeptide hormone in association with its specific receptor. PMID- 6280963 TI - Lipoprotein-binding sites in human corpus luteum membrane fractions. AB - In a previous report evidence was presented that plasma low density lipoprotein (LDL), but not high density lipoprotein (HDL), is the major source of cholesterol used by the human corpus luteum for progesterone biosynthesis and, that plasma LDL is taken up by corpus luteum tissues via a receptor-mediated endocytotic process. Using membrane fractions prepared from fresh corpus luteum tissue obtained from nine women at various phases of the menstrual cycle, we conducted the present investigation to characterize lipoprotein-binding sites and changes in such binding sites that might occur throughout the ovarian cycle. High affinity, low capacity binding sites for [125I]iodo-LDL and also for [125I]iodo HDL were detected in membrane fractions prepared from fresh corpus luteum tissue. The interaction of [125I]iodo-LDL with the high affinity binding sites in fresh corpus luteum membrane fractions was prevented by incubation with heparin. Also, the binding capacity of corpus luteum membrane fractions for [125I]iodo-LDL was reduced by treatment with pronase. On the other hand, the specific binding capacity for [125I]iodo-HDL was unaffected by pronase treatment. The binding capacity for [125I]iodo-LDL in membrane fractions prepared from midluteal phase corpus luteum was significantly greater than that of membrane fractions from tissue obtained at any other phase of the cycle, a finding that suggests that changes in progesterone biosynthesis throughout the cycle are positively correlated with changes in the numbers of binding sites for LDL in the corpus luteum. PMID- 6280965 TI - Thyroid hormone specifically inhibits prolactin synthesis and decreases prolactin messenger ribonucleic acid levels in cultured pituitary cells. AB - Electrophoretic analysis of soluble proteins from pituitary cells pulse labeled with [35S]methionine demonstrated that 10 nM T3 inhibited PRL synthesis, but did not affect the synthesis of most other pituitary proteins. The effects of T3 were somewhat slow, requiring about 3 days for a 50% reduction in PRL synthesis. PRL synthesis slowly returned toward control levels after the removal of T3 from the culture medium. In serum-free medium, a concentration of about 0.6 nM T3 was required for half-maximal inhibition of PRL synthesis. In medium containing 5% fetal calf serum, only slightly higher concentrations of T3 were required to inhibit PRL synthesis. The Kd for the binding of [125]T3 to pituitary cell nuclei was 0.2 nM. Analysis of PRL mRNA levels by hybridization of total cellular RNA to PRL cDNA demonstrated that there was a good correspondence between T3 effects on PRL synthesis and PRL mRNA. These findings demonstrate that T3 can specifically inhibit PRL synthesis and PRL mRNA levels in cultured pituitary cells and suggest that T3 may have a physiological role in the regulation of PRL synthesis. PMID- 6280964 TI - Mechanism of the inhibitory action of epidermal growth factor on testicular androgen biosynthesis in vitro. PMID- 6280966 TI - Stimulation by follicle-stimulating hormone of synthesis of lactate by Sertoli cells from rat testis. AB - Sertoli cells from rats aged 16 days were cultured in defined medium for 2 days and then treated by addition of fresh medium containing various hormones (treated) or saline (control). The concentration of lactate in the medium was measured as a function of time. The production of lactate measured under these conditions was increased by addition of FSH to the medium. For NIH FSH (13 and 14), ED50 for stimulation of lactate production was approximately 0.05 micrograms/ml. Stimulation was also seen with LH and TSH (ED50, 0.8 micrograms/ml for both hormones). Reasons are given for believing that TSH may possess the inherent capacity to stimulate production of lactate in contrast to LH, which appears to act only by way of contaminating traces of FSH. Dibutyryl cAMP also stimulates lactate production by Sertoli cells. Other hormones tested, including androgens, were without effect. When production of lactate by Sertoli cells was compared with the maximal consumption by spermatids in vitro, it became apparent that Sertoli cells may provide the major source of metabolic substrate for spermatids in the form of lactate. Stimulation of lactate production by FSH was inhibited by puromycin, cycloheximide, and actinomycin D. Evidently this response requires synthesis of new protein and RNA. This effect of FSH may be important in the regulation of spermatogenesis. PMID- 6280967 TI - Rapid decreases in adrenal and plasma corticosterone concentrations after drinking are not mediated by changes in plasma adrenocorticotropin concentration. AB - Rats given water to drink only during a brief daily period respond to water presentation with a rapid decline in plasma corticosterone concentration. To determine whether this response is consequent to a decrease in plasma ACTH concentration or whether it reflects a sudden reduction in adrenal responsiveness to ACTH, we allowed rats access to water for 2 h/day at lights on and measured plasma ACTH and adrenal and plasma corticosterone concentrations at 3- or 5-min intervals after the onset of drinking. Adrenal and plasma corticosterone concentrations decreased significantly within 2-3 min after water presentation in the absence of concomitant changes in plasma ACTH concentration. The effect was apparent by 5 days after initiating the restricted drinking schedule and became stronger with time up to 21 days. Further characterization of the response showed that the in vitro corticosterone secretion of adrenals removed at intervals after water presentation followed the same pattern as the in vivo concentration. When empty water bottles were presented, plasma ACTH and corticosterone concentrations increased. Neither adrenal medullary function nor plasma renin concentration was found to be associated with the decline in adrenal responsiveness to ACTH after drinking. Hemisection of the spinal cord of unilaterally adrenalectomized rats attenuated the corticosterone response regardless of whether the hemisection was contralateral or ipsilateral to the remaining adrenal. These data suggest that the decreases in plasma and adrenal corticosterone concentration occurring after drinking in water-restricted rats are not dependent on changes in plasma ACTH concentrations, but may be related to changes in adrenal blood flow, steroid metabolism and distribution, or neural input to the adrenal cortex. PMID- 6280969 TI - Effect of erythrosine (2',4',5',7'-tetraiodofluorescein) on the metabolism of thyroxine in rat liver. PMID- 6280968 TI - Low protein-high carbohydrate diet induces alterations in the serum thyronine binding proteins in the rat. AB - The serum T3 concentration was increased in 8-week-old lean Zucker rats fed a low protein-high carbohydrate diet for 2 weeks. This increase was secondary to the generation of a binding protein migrating in the postalbumin zone in polyacrylamide gel electrophoresis employing 125I-labeled T3 and is termed rat thyronine-binding globulin. The presence of this T3-binding protein in serum resulted in a marked decrease in the percent free T3 assessed by equilibrium dialysis and a normal free T3 concentration. An increase in the binding of T4 in the postalbumin zone was also observed, but no changes in the dialyzable fraction of T4 or the total and free T4 concentrations occurred. In contrast to these findings in lean Zucker rats fed the low protein-high carbohydrate diet, no change in the pattern of 125I-labeled T3 and T4 binding, the dialyzable fraction of T3 or T4, or total and free T3 or T4 concentrations were observed in the obese Zucker rats fed this diet. The present findings suggest that diet-induced alterations in thyroid hormone-binding proteins must be considered in the interpretation of data which involve alterations in total thyroid hormone concentrations in serum and their role in affecting tissue metabolism. PMID- 6280970 TI - Structural analysis of rodent growth hormone genes: application to genetic forms of hypopituitarism. AB - Analysis of GH gene structure in the mouse permits evolutionary comparisons with GH gene structure in the rat and provides information about the mechanism responsible for heritable deficiencies of anterior pituitary hormones. The little mutation in mice is analogous to autosomal recessive, isolated, partial deficiency of GH in man, whereas the Snell dwarf mutation is a model for autosomal recessive deficiency of GH, TSH, and PRL. Mouse cellular DNA was digested with the restriction enzymes Eco RI, Bam HI, Bgl II, Hind III, and Kpn I, singly and in combination. Gene sequences containing coding information for GH were detected by hybridization to a radiolabeled recombinant DNA probe complementary to a rat GH mRNA. The results of genomic restriction analysis indicate that there is a single type of mouse GH gene sequence per haploid genome with a length equal to or less than 32000 base pairs. The distances separating 6 base restriction sites close to the mouse GH gene differ from those close to the rat GH gene. A Kpn I site in the codons for amino acids 103 and 104 of rat GH is absent in the mouse gene. Restriction patterns of DNA from little and Snell dwarf mice did not differ from those of normal mice, indicating that these mutations do not involve deletions of the mouse GH gene. PMID- 6280971 TI - Storage of anterior lobe adrenocorticotropin in corticotropes and a subpopulation of gonadotropes during the stress-nonresponsive period in the neonatal male rat. AB - The neonatal rat exhibits a stress-nonresponsive period during days 4-11 of postnatal development. The role and response of the anterior lobe corticotrope was studied with immunocytochemical stains for ACTH on fixed embedded pituitaries from male rats 2-21 days of age. Serially section fields were stained for beta chains of LH or FSH to test for joint storage of ACTH and one or both gonadotropins. Cell counts on semithin sections were used to determine the frequency of stained corticotropes in the developing pituitary. The 2-day-old rats had twice as many corticotropes (17.6%) as the adults (8.1%). During the stress-nonresponsive period, the frequency of corticotropes declined to 6.4% of the population. This was followed by a recovery to 16.9% at 15 days of age. The serial fields showed that stellate cells containing only ACTH declined sharply (by 95%) to less than 1% of the pituitary cell population during the first week of postnatal life. Cells containing ACTH and both gonadotropins predominated in the corticotrope population and were 4-6% of the pituitary cell populations during this time period. In the 15-day-old rats, the corticotropes included cells storing ACTH alone (5%) and the cells storing ACTH and both gonadotropins (9%). Throughout development, cells storing ACTH alone were distinguished by their intense staining, stellate shape, and peripheral granules. Cells storing ACTH and gonadotropins were stellate or ovoid and often resembled maturing gonadotropes. We hypothesize that this second group of cells serves a function related to adrenal-gonadal maturation, or they may be stem cells, abundant during development and present in relatively low percentages (1-3%) in adult rats. PMID- 6280972 TI - The importance of the hypothalamic lateral retrochiasmatic area in the control of adrenocorticotropin and thyrotropin secretion. AB - Anterolateral hypothalamic deafferentiation was made in rats to explore the importance of the neural pathways through the lateral retrochiasmatic area (RCAL) in the regulation of ACTH and TSH secretion. In rats with complete bilateral RCAL transection, pituitary-adrenal function was altered in the following respects compared to sham-operated controls. 1) Basal plasma ACTH, corticosterone (B), and adrenal weight were depressed. 2) Plasma ACTH and B elevation in response to 3 min ether inhalation were markedly decreased or abolished. 3) Insulin-induced hypoglycemia produced no or little plasma B elevation. 4) Lysine-vasopressin was significantly less effective in inducing pituitary-adrenal activation. Reductions in plasma ACTH and B concentrations and adrenal weight were correlated with the completeness of the RCAL transection. The plasma TSH concentration was lower in the deafferented rats than in the controls regardless of the completeness of the cut at the RCAL, indicating that the neural pathways traversing this area do not possess a critical importance for the regulation of TSH if the rest of the hypothalamus is deafferented anterolaterally. We conclude that intact neural connections between the medial basal hypothalamus and the central nervous system at the RCAL are essential for the maintenance of normal hypothalamic-pituitary adrenal function. PMID- 6280973 TI - Regulation of thyrotropin-releasing hormone receptors and responses by L triiodothyronine in dispersed rat pituitary cell cultures. AB - The effects of physiological concentrations of L-T3 (T3) were examined in dispersed cell cultures of pituitaries obtained from 10- to 12-day-old rats. T3 inhibited TSH secretion by 50% and blunted the TSH response to TRH. The PRL response to TRH was also inhibited by T3, and GH secretion was increased 2-fold. These responses were half-maximal at 0.1 nM added T3 in medium supplemented with 10% hypothyroid calf serum, corresponding to a free T3 concentration of 5 pM. In the presence or absence of added T3, TRH effects were half-maximal at 0.5-3 nM, and T3 suppression was not overcome by high concentrations of TRH (up to 1 microM). Maximal inhibition of TSH responses to TRH occurred when cultures were preincubated with thyroid hormone for 24 h; a significant effect was observed after 8 h. The specific binding of [3H]TRH to dispersed rat pituitary cells was decreased 55-70% by T3 in a dose-dependent manner. Inhibition of TSH secretion by T3 was reversible within 24 h, and the fraction of thyrotrophs in the cultures (0.22) was not altered by T3 over the course of the experiments. The results demonstrate that physiological concentrations of T3 regulate TSH and PRL responses to TRH and control TRH receptor levels by a direct action on normal rat pituitary cells. PMID- 6280974 TI - Serum binding of triiodothyronine: theoretical and practical implications for in vitro triiodothyronine uptake. PMID- 6280975 TI - Responses of isolated guinea-pig adrenal cells to ACTH and pro-opiocortin-derived peptides. AB - A comparison of the responses of isolated guinea-pig adrenal cells to ACTH and pro-opiocortin-derived peptides was carried out by measuring cortisol, aldosterone, androstenedione and dehydroepiandrosterone production. With concentrations below 10,000 pg/ml, no steroidogenic activity was found in response to either beta-LPH, gamma-LPH, gamma 3-MSH or the 16K fragment, whether assayed alone or in association with ACTH. At concentrations above 10,000 pg/ml, gamma-LPH (100 ng), the 16K fragment (100 ng) and beta-endorphin (500 ng) proved to be totally inactive. beta-LPH from 25 to 250 ng, however, exhibited a significant though slight stimulatory effect on cortisol, aldosterone and androstenedione production. Its effectiveness on aldosterone production was especially marked, but the extent of the response was modest in view of the concentrations used. PMID- 6280976 TI - Dopamine regulates canine plasma beta-endorphin-immunoreactivity levels. AB - Intravenous administration of the dopamine antagonists domperidone and metoclopramide elevates canine plasma beta-endorphin immunoreactivity. Pretreatment with dopamine, but not dexamethasone, inhibits this effect. Sephadex G-50 chromatography shows that this response is due to beta-endorphin-like peptides, not beta-lipotropin. These findings indicate that pituitary beta endorphin secretion is tonically inhibited by dopamine and suggest that the intermediate lobe is the source of this secretion. PMID- 6280977 TI - The pituitary-adrenocortical response to hemorrhage depends on the time of day. AB - The response of plasma ACTH and the secretory response of cortisol to moderate hemorrhage were determined in awake dogs in the morning (AM) and evening (PM). Whereas the magnitudes of the response of ACTH were similar in the AM and PM, the magnitude of the secretory response of cortisol was significantly greater in the AM compared to that in the PM. At both times of day, heart rate, mean arterial pressure, and secretory rates of epinephrine after hemorrhage were similar. These findings suggest that AM-PM differences in stimuli produced by moderate hemorrhage cannot explain the differences in the secretion of cortisol. Instead, AM-PM changes in adrenocortical sensitivity to endogenous ACTH after hemorrhage determine the AM-PM differences in the secretion of cortisol. PMID- 6280978 TI - Chronic haloperidol treatment increases the level of in vitro translatable messenger ribonucleic acid coding for the beta-endorphin/adrenocorticotropin precursor proopiomelanocortin in the pars intermedia of the rat pituitary. AB - Chronic treatment of rats with haloperidol (1.5 mg/kg, once daily) over a period of 7--21 days resulted in a 80--100% increase in the tissue levels of immunoreactive beta-endorphin and in the in vitro release of immunoreactive beta endorphin from the neurointermediate pituitary. Incorporation of [3H]phenylalanine into isolated neurointermediate pituitaries of haloperidol treated rats revealed an increase in the amount of label incorporated into the beta-endorphin/ACTH precursor proopiomelanocortin (POMC) to a similar extent (about 80%) but had essentially no effect on the conversion of the precursor into beta-lipotropin and beta-endorphin. Extraction of messenger (m) RNA from neurointermediate pituitaries followed by cell-free translation in a reticulocyte system showed an increase in the total level of translatable mRNA (about 25%). The content of translatable mRNA coding for POMC, however, was increased by 100 150%. Time-course studies revealed a parallelism between the effect of haloperidol on the level of in vitro translatable mRNA coding for POMC and the ability of the drug to increase the concentrations of beta-endorphin in the neurointermediate pituitary. A complete reversal of the effects of haloperidol was seen 2 weeks after discontinuation of the drug. These findings suggest that the chronic blockade of dopaminergic receptors by haloperidol causes a reversible increase in the beta-endorphin biosynthesis in the rat intermediate pituitary at the pretranslational level by markedly increasing the level of translatable mRNA coding for POMC. PMID- 6280979 TI - Somatostatin-like immunoactivity and biological activity is present in Tetrahymena pyriformis, a ciliated protozoan. PMID- 6280980 TI - Vitamin D metabolism in pregnant rabbits: differences between the maternal and fetal response to administration of large amounts of vitamin D3. AB - Maternal and fetal metabolism of vitamin D was examined in term pregnant rabbits fed a normal diet and in those supplemented with a large amount of vitamin D3. Term pregnant rabbits (27--30 days of gestation) fed the normal diet showed lower levels of plasma calcium, 25-hydroxyvitamin D3 (250HD3), and 24,25 dihydroxyvitamin D3 [24,25-(OH)2D3] and higher plasma 1 alpha, 25 dihydroxyvitamin D3 [1 alpha, 25-(OH)2D3] levels than age-matched nonpregnant female rabbits. Kidney homogenates from pregnant rabbits produced mainly 1 alpha 25-(OH)2D3, while those from nonpregnant animals produced 24,25-(OH)2D3 primarily. Plasma concentrations of calcium and phosphorus were significantly higher in fetuses than in mothers. Plasma levels of 250HD3 and 24,25-(OH)2D3 in fetuses were almost identical to those in mothers, whereas 1 alpha,25-(OH)2D3 levels in plasma were significantly higher in mothers than in their fetuses. A daily administration of 650 nmol vitamin D3 for 3 days to term pregnant rabbits caused a significant increase in calcium, phosphorus, 25OHD3, and 24,25-(OH)2D3 in maternal plasma, and in 25OHD3 and 24,25-(OH)2D3, but not calcium and phosphorus in fetal plasma. Treatment with large amounts of vitamin D3 also induced a marked suppression of 1 alpha-hydroxylase activity and a concomitant increase of 24-hydroxylase activity in the maternal but not in the fetal kidney. Plasma concentrations of 1 alpha,25-(OH)2D3 were not affected by treatment with large amounts of vitamin D3 in either the fetuses or the mothers. These results clearly indicate that the renal 25OHD3 metabolism in the fetus is regulated independently of that in the mother. PMID- 6280981 TI - Adenosine 3',5'-monophosphate derivatives increase prolactin synthesis and prolactin messenger ribonucleic acid levels in ergocryptine-treated pituitary cells. PMID- 6280982 TI - Low density lipoprotein binding and de novo synthesis of cholesterol in the neocortex and fetal zones of the human fetal adrenal gland. AB - The binding of low density lipoprotein (LDL) and the de novo synthesis of cholesterol in separated zones of human fetal adrenal (HFA) tissues were investigated. The number of LDL-binding sites was 2-fold greater in membrane fractions prepared from fresh fetal zone tissue than in those from neocortex tissue. The binding capacity for LDL in fetal zone and neocortex membrane preparations of HFA tissues maintained in culture in the presence of ACTH was 2 fold greater than that in membrane fractions of control tissues. The rates of de novo synthesis of cholesterol also were determined in separated zones of HFA tissue by measuring the specific activity of 3-hydroxy-3-methylglutaryl coenzyme A reductase in microsomal fractions prepared from HFA tissues and by determining the rate of incorporation of tritium from [3H]water into cholesterol in HFA tissue fragments. The rate of de novo synthesis of cholesterol in fresh fetal zone tissue was twice that in neocortex tissue as estimated by these methods. When separated zones of HFA tissue were maintained in culture in the presence or absence of ACTH, the rates of de novo synthesis, as determined by the rate of incorporation of tritium from [3H]water into cholesterol, were stimulated to a similar extent by ACTH in both fetal zone and neocortex tissues. However, the specific activity of 3-hydroxy-3-methylglutaryl coenzyme A reductase was increased to a greater extent by ACTH pretreatment in neocortex tissues than in fetal zone tissues. In summary, fetal zone tissues of the HFA gland have a larger number of LDL-binding sites and higher rates of de novo synthesis of cholesterol than do neocortex tissues, and ACTH stimulates LDL binding and de novo synthesis of cholesterol in both zones of the HFA gland. PMID- 6280983 TI - Calcium-dependent regulation of aldosterone production in isolated adrenal glomerulosa cells: effects of the ionophore A-23187. AB - The effect of the calcium ionophore A-23187 on the regulation of aldosterone synthesis was investigated in isolated rat adrenal glomerulosa cells. Over the concentration range of 0.1--1.0 microM, A-23187 increased aldosterone production in a dose-dependent manner, with half-maximal stimulation at approximately 0.3 microM. Stimulation of aldosterone production by A-23187 was dependent on the concentration of extracellular calcium and was not accompanied by detectable increases in cAMP production. In contrast to the stimulation of steroidogenesis elicited by A-23187 alone, the ionophore significantly inhibited the aldosterone responses to angiotensin II and potassium. In angiotensin II-stimulated cells, 0.2 microM A-23187 caused half-maximal inhibition of aldosterone production, and 2 microM A-23187 inhibited aldosterone production by about 70%. The ionophore did not affect the binding of angiotensin II to isolated glomerulosa cells and did not change the sensitivity of the cells to either angiotensin II or potassium. However, A-23187 reduced the total amount of steroid produced in response to these stimuli, most markedly when the extracellular calcium concentration was less than 0.4 mM. These results indicate that although A-23187 partially inhibits the steroidogenic responses to angiotensin II and potassium, the increases in cytosolic calcium caused by submicromolar ionophore concentrations result in stimulation of aldosterone production by adrenal zona glomerulosa cells. PMID- 6280985 TI - Receptors for vasoactive intestinal polypeptide on crude smooth muscle membranes from porcine uterus. PMID- 6280984 TI - Adenosine receptor-mediated accumulation of adenosine 3',5'-monophosphate in guinea pig thyroid tissue. AB - The cAMP content of guinea pig thyroid fragments was increased 2- to 3-fold by adenosine analogs, 5'-N-ethylcarboxamide adenosine (NECA), the most potent, caused half-maximal stimulation of 0.7 microM, whereas N6-phenylisopropyl adenosine was much less potent (half-maximal stimulation at 20 microM). Exogenous adenosine by itself was slightly active, and its activity was increased in the presence of adenosine deaminase or transport inhibitors. Although adenosine analogs are partial agonists compared to TSH or the diterpene cyclase activator forskolin, the fractional rate of cAMP accumulation is greater for NECA than for TSH. The stimulatory effect of NECA was antagonized by theophylline. These effects of adenosine and its analogs on cAMP formation in intact thyroid tissue are characteristic of stimulatory adenosine receptors. PMID- 6280986 TI - Gonadotropin receptors of the primate corpus luteum. II. Changes in available luteinizing hormone- and chorionic gonadotropin-binding sites in macaque luteal membranes during the nonfertile menstrual cycle. PMID- 6280987 TI - Differential effects of dithiothreitol and iodoacetamide on corticotropin releasing factor (CRF) activity of bovine hypothalamic CRFs and vasopressin. AB - Various fractions were tested in vivo for corticotropin-releasing factor (CRF) activity after Sephadex G-100 fractionation of 0.1-N HCl extracts of bovine hypophyseal stalk or cerebral cortex. Female rats pretreated with chlorpromazine, morphine, and Nembutal were used for CRF assay. CRF-A (void volume fractions; big CRF), CRF-B (Kav = 0.583), and CRF-C (salt volume fractions) of bovine hypophyseal stalk and lysine or arginine vasopressin all induced clear-cut stimulation of ACTH and corticosterone in the assay rat, whereas they were ineffective in acutely hypophysectomized rats. Control fractions purified from bovine cerebral cortex had no CRF activity. Treatment of arginine and lysine vasopressin and CRF-C with dithiothreitol and iodoacetamide completely abolished their CRF activity, whereas the CRF activities of CRF-A and CRF-B were unaltered by these treatments. Treatment with iodoacetamide alone had no effect on the CRF activity of any of these substances. Fractionation of either CRF-C or arginine vasopressin on Sephadex G-15 yielded a CRF-active peak at a Kav of 0.35. We conclude that 1) three different forms of CRF exist in bovine hypophyseal stalk; 2) CRF-A and CRF-B are unrelated to vasopressin and require neither a disulfide bond(s) nor a sulfhydryl group(s) for their CRF activity; 3) reduction of the disulfide bond of vasopressin destroys both CRF and antidiuretic activities; 4) CRF-C requires an intact disulfide bond(s) for its CRF activity and is likely to be either vasopressin itself or a substance closely related to vasopressin; and 5) CRF-B is likely to be the physiologically important form of bovine CRF. PMID- 6280988 TI - Adrenocorticotropin, and corticosterone secretion in Brattleboro rats. AB - Brattleboro rats which lack endogenous vasopressin have been used to study the role of vasopressin as a corticotropin-releasing factor. Plasma ACTH, beta endorphin, and corticosterone were measured by RIA in male and female Long-Evans and Brattleboro rats under the following conditions: unstressed, after ether stress, after nicotine injection, and after adrenalectomy. A significant reduction in the ACTH, beta-endorphin, and corticosterone responses to the different experimental procedures was observed in the Brattleboro rats. However, in this strain of rats, a significant increase in the release of all three hormones was obtained, suggesting that vasopressin has only a synergistic role in the regulation of their secretion. PMID- 6280989 TI - A comparison between the effects of growth hormone on prolactin receptors and estrogen receptors in rat liver. AB - Human GH (hGH) increased hepatic PRL and estrogen receptors in hypophysectomized (Hx) female rats after continuous infusion of the hormone (5 micrograms/h) using osmotic minipumps, but not after the infrequent administration of the same daily dose (120 micrograms) of the hormone by sc injections (60 micrograms/12 h). The effects of hGH on body weight (an increase) and tibial epiphyseal zones (a widening) were observed regardless of the mode of hGH administration. Thus, it would seem as if the mode of administration of hGH is of importance for the type of biological effect exerted by the hormone. Furthermore, rGH was effective in increasing both PRL and estrogen receptors in the liver of Hx ovariectomized (HxOx) rats, whereas rat PRL (rPRL) was much less efficient. Finally, rat GH (rGH) suppressed the concentration of a nonreceptor estrogen-binding protein in the liver of HxOx rats; the protein occurred at a low concentration in Ox rats and was markedly increased after hypophysectomy rPRL was inefficient in modulating the concentration of estrogen-binding protein. Thus, rGH, but not rPRL, seems to regulate both sexually differentiated (PRL receptors and estrogen binding protein) and sexually nondifferentiated (estrogen receptor) functions in the rat liver. The mechanism behind this dual effect of rGH is obscure, but may possibly be related to the presence of isohormones of rGH with different effects on the liver. PMID- 6280990 TI - ACTH and beta-endorphin-related peptides are present in multiple sites in the reproductive tract of the male rat. PMID- 6280991 TI - Anaphase aberrations: a measure of genotoxicity in mutagen-treated fish cells. AB - Rainbow trout gonad cells (RTG-2) were cultured for various lengths of time in the presence of several classes of known mutagenic chemicals and several related compounds that possessed no known mutagenic/carcinogenic activity. During the course of exposure the cells were examined for the presence of abnormalities in the chromosome arrangement of anaphase figures during mitosis. Untreated and solvent-treated (dimethylsulfoxide-treated) cells exhibited a background abnormality rate of 12% with only minor chromosomal defects being observed. This was also true for those cells exposed to naphthol and anthracene, two chemicals with no proven mutagenic or carcinogenic activity. Conversely, significant increases in the frequency of anaphase aberrations were produced in cells treated with N-methyl-N'-nitro-N-nitrosoguanidine, benzo(a)pyrene, 9-aminoacridine and mitomycin-C. These abnormalities were also far more complex and extensive than those observed in the control and nonmutagen-treated cells. Many species of fish have extremely small and numerous chromosomes, making resolution of chromosome defects such as sister chromatid exchange and deletions more difficult than in most mammalian diploid cells, which generally have larger and fewer chromosomes. Examination of cells during anaphase eliminates the need to observe each chromosome separately as well as the need to produce well-spread metaphase chromosomes. Since the sensitivity of anaphase aberrations to known mutagenic/carcinogenic compounds appears to be quite high in trout cells and since hundreds of suitable cells are available for analysis, this may be an appropriate alternative or addition to some of the more standard chromosome macrolesion tests developed in mammalian systems. PMID- 6280992 TI - Kinetic mechanism of the exchanges catalysed by the adenine-nucleotide carrier. AB - Initial rates of the exchange ADPin/ADPout catalysed by the adenine-nucleotide carrier of rat-heart mitochondria have been studied under conditions where internal and external ADP may be varied. The initial rate was measured within 1 s by the carboxyatractyloside-stop method, using a rapid-mixing technique. The double-reciprocal plots v0(-1) versus [ADP]out-1 at different internal-ADP concentrations and v0(-1) versus [ADP]in-1 at different external-ADP concentrations exhibit straight-line relationships having a common point of intersection on the axis of ordinates. These results demonstrate the essential role of a ternary complex and thus exclude the ping-pong mechanism generally accepted. The kinetic equation implies a strong positive cooperativity in the binding of the two substrates. Two models are proposed: (a) the ternary complex performs the exchange and the transport of the substrates in a single step; (b) the carrier is mobile and transports the substrates one by one, the formation of a ternary complex being needed to release the first product. PMID- 6280993 TI - The catalytic mechanism of glutamyl-tRNA synthetase of Escherichia coli. Evidence for a two-step aminoacylation pathway, and study of the reactivity of the intermediate complex. PMID- 6280994 TI - Nucleotide sequence of the simian virus 40 HindII + III restriction fragment A (second part of the T antigen gene). AB - The nucleotide sequence of the second part of the simian virus 40 DNA HindII + III restriction fragment A is presented. The sequence extends from map position 0.533 to 0.424 and together with the first part of Hind-A [Volckaert et al. Proc. Natl Acad. Sci. U.S.A. 75, 2160--2164 (1978)] completes the total hind-A sequence, comprising 1169 base pairs. The second half of Hind-A includes the region corresponding to the second splicing boundary common to small tumor antigen (small-t) and large tumor antigen (large-T) mRNA and it contains coding information for an internal portion of large-T antigen. Two similar secondary structures of reasonable thermodynamic stability can be proposed for the nucleotide sequence of the pre-mRNA corresponding to the region reported here. Their possible relevance to the splicing of the SV40 early mRNAs is discussed. The deduced amino acid sequence is 188 residues long and contains a Lys-Lys-Lys Arg-Lys-stretch which may be involved in the DNA binding capacity of large-T. A presumptive phosphorylation site is also present. PMID- 6280995 TI - Nucleotide sequence of the simian virus 40 HindII + III restriction fragment I (fourth part of the T antigen gene). AB - The HindII + III restriction fragment I (Hind-I) from simian virus 40 DNA represents 4.96% of the genome and maps in the early transcription region. Hind-I is an internal segment of the A gene and its information is expressed as part of the early 19-S mRNA, which codes for T antigen. We report here the nucleotide sequence of the 259-base-pair Hind-I fragment. The sequence was determined and confirmed by RNA and DNA sequencing methods: by analysis of oligonucleotides resulting from T1 and pancreatic RNase digestion of labeled RNA transcribed from SV40 DNA with Escherichia coli DNA-dependent RNA polymerase, by partial degradation of RNA transcripts with snake venom phosphodiesterase, and by base specific chemical degradation of 5'-end-labeled subfragments of Hind-I according to the procedure of Maxam and Gilbert. Multiple triplets corresponding to termination codons occur in two of the three reading frames of the DNA strand that has the same polarity as early mRNA. The open reading frame connects in phase with the one of the Hind fragments flanking Hind-I, and the amino acid sequence specified by Hind-I lies in the middle part of the large-T antigen. Some features of the primary nucleotide sequence and of early transcription are discussed. PMID- 6280996 TI - Nucleotide sequence of the Hind-I-proximal part of simian virus 40 HindII + III restriction fragment B (fifth part of the T antigen gene). AB - The nucleotide sequence of the simian virus 40 DNA segment that lies between the HindIII restriction endonuclease cleavage site at map position 0.324 and the AtuI cleavage site at 0.261 has been determined by the base-specific partial chemical degradation procedure of Maxam and Gilbert. This region comprises 335 base pairs and represents 6.4% of the total SV40 genome and 41% of the restriction fragment Hind-B. It connects in the clockwise direction to the restriction fragment Hind-I (described in the preceding paper). Hind-B is situated in the second half of the early transcription region of the SV40 genome and encodes information (including the termination signal) for the early protein large-T antigen. in the part of Hind-B described here, the DNA strand that has the same polarity as early 19-S mRNA defines only one open reading frame for translation; the other two are blocked by multiple triplets corresponding to termination codons. The open reading frame is part of one that runs throughout much of the early region: from the second splicing boundary of the large-T gene (position 0.534) to the information for the large-T stop signal (position 0.174) near the Hind-B/Hind-G junction, and which together with the non-contiguous DNA segment from position 0.65 (site of the information for the large-T start signal) to 0.69 (the first splicing boundary) codes for the entire large-T antigen. PMID- 6280997 TI - Nucleotide sequence of simian virus 40 DNA: structure of the middle segment of the HindII + III restriction fragment B (sixth part of the T antigen gene) and codon usage. AB - We report here the nucleotide sequence of the simian virus 40 DNA region that lies between the EcoRII restriction endonuclease cleavage sites at map positions 0.214 and 0.281. The sequence was determined by partial chemical degradation of terminally labeled DNA fragments according to the procedure of Maxam and Gilbert. This region represents 6.7% of the SV40 genome and is located in the middle of HindII + III restriction fragment B. It is expressed as part of the early 19-S messenger RNA, which codes for the large-T antigen protein. Only one open reading frame for translation can be deduced from the message strand of the DNA and this reading frame connects in phase with the one of both neighboring fragments. This publication is the last in a series of papers about the T-antigen gene, and several properties of this gene and its product are discussed. The non-randomness of codon usage is similar to that previously discussed for the late part of the genome. Moreover, it appears that the choice of a third letter can be determined by the nature of the following codon; some codons which start with a pyrimidine are almost never preceded by an adenosine and some ANN-type codons are almost never preceded by a guanosine. PMID- 6280998 TI - Localization of the binding sites of prokaryotic and eukaryotic RNA polymerases on simian virus 40 DNA. AB - The binding sites of calf thymus RNA polymerase (B) II, wheat germ RNA polymerase B and of the Escherichia coli RNA polymerase were mapped on the simian virus 40 genome by observation of enzyme-linear DNA complexes by electron microscopy. Three to four major sites and several minor sites are observed for each enzyme; common binding sites for the three enzymes are found in positions 0.17, 0.53 and 0.90 of the viral physical map. Initiation complexes with these enzymes can be stabilized with specific ribodinucleotides and a single ribonucleoside triphosphate. Whereas ApA and ATP greatly enhances the binding of the E. coli enzyme at position 0.17, they stabilize the binding of the eukaryotic enzyme at many sites, some of them located in close proximity of the origin of replication. PMID- 6280999 TI - Structural studies on heparan sulphates. Characterization of oligosaccharides; obtained by periodate oxidation and alkaline elimination. AB - Three heparan sulphate fractions were subjected to degradation by periodate oxidation and alkaline elimination. The starting materials were one low-sulphated fraction rich in glucuronic acid and N-acetylglucosamine (I), one fraction (IV) that was medium-sulphated, contained similar proportions of N-acetyl and N sulphate as well as of glucuronic and iduronic acid and one fraction (V) that was oversulphated, contained an excess of N-sulphate and was rich in iduronic acid. Selective periodate oxidation of glucuronic acid residues within N-acetylated regions followed by scission in alkali yielded three categories of fragments that were isolated by gel and ion-exchange chromatography. (a) N-Acetylglucosamine-R compounds where R is the remnant of an oxidised and degraded glucuronic acid residue. (b) Medium-sized oligosaccharides, of the general structure glucosamine (glycuronic acid-glucosamine)n-R where n = 1--4, contained largely glucuronic acid associated with glucosamines that were N-sulphated, N-acetylated or unsubstituted. In saccharides where n = 4 indications of an alternating arrangement of N-sulphate and N-acetyl groups as well as of iduronic and glucuronic acid residues were obtained. The glucuronic acid residues of the oligosaccharide fragments were susceptible to reoxidation with periodate. The smaller saccharides were not depolymerised by HNO2 despite the presence of N sulphate groups. (c) Longer oligosaccharide fragments (n greater than or equal to 5) that were highly sulphated and contained heparin-like repeating units, i.e. iduronic acid--glucosamine-N-sulphate with ester-sulphate on both sugars. These oligosaccharides were susceptible to deaminative cleavage. PMID- 6281000 TI - Properties of the interaction of the sodium channel with permeant monovalent cations. AB - The use of sea anemone toxin, veratridine and scorpion toxin which specifically interact with the gating system of the sodium channel and maintain the channel in an open conformation has permitted a study of the mechanism of transport of monovalent cations through the selectivity filter of this channel. The initial rate of 22Na+ influx through the tetrodotoxin-sensitive Na+ channels of excitable cells is dependent upon the external concentrations of Na+ and Na+-substitutes with the following properties. (a) It is saturable at high Na+ concentrations and increases with the external Na+ concentration in a cooperative manner (nH = 1.6). (b) At low external Na+ concentrations (1 mM), it is activated and then inhibited by increasing external concentrations of monovalent cations such as Li+, guanidinium, hydrazinium, hydroxylamine and K+. The activating effect of these cations disappears at higher external Na+ concentrations (10 mM). The experimental data are consistent with a model involving at least two allosteric cation-binding sites per Na+ channel. The binding of monovalent cations to Na+ sites is characterized by a high positive homotropic cooperativity. Most of the work describes the properties of the Na+ channel in neuroblastoma cells. The mechanism has also been shown to be valid for excitable cells of other types and origins. PMID- 6281001 TI - Isoleucyl-tRNA synthetase from Baker's yeast. Action of ATP analogs in pyrophosphate exchange and aminoacylation, two pathways of the aminoacylation depending on concentration of pyrophosphate. AB - The order of substrate addition to isoleucyl-tRNA synthetase from baker's yeast has been investigated by steady-state kinetics with inhibition by four different inhibiting ATP analogs acting competitively, uncompetitively and noncompetitively with respect to ATP, namely purineriboside (= nebularin), 3'-deoxy-adenosine (= cordycepin), 8-amino-adenosine and 8-azido-adenosine 5'-triphosphates. The inhibition studies were done in the aminoacylation and in the pyrophosphate exchange reaction, the aminoacylation was investigated in the absence and presence of inorganic pyrophosphatase. Additionally, bisubstrate kinetics and product inhibition studies were carried out. The inhibition patterns indicate a multisite system with a minimum number of two sites for each of the substrates. The results of the pyrophosphate exchange studies are consistent with formation of E . Ile-AMP . ATP . Ile complexes by random addition of one ATP and one isoleucine molecule, followed by adenylate formation, subsequent release of pyrophosphate and random addition of a second molecule of ATP and isoleucine. For the aminoacylation in the absence of pyrophosphatase an ordered ter-ter mechanism is postulated; in the presence of pyrophosphatase the mechanism is random bi-uni uni-bi ping-pong. Both the pyrophosphate and the analogs of this compound such as imidodiphosphate or methylenediphosphonate can induce the enzyme to act in the ter-ter mechanism. PMID- 6281003 TI - A comparison of purified poly(ADP-ribose) polymerases from Ehrlich ascites tumor cells, pig thymus, and HeLa S3 cells. AB - Poly(ADP-ribose) polymerases from Ehrlich ascites tumor cells, pig thymus, and HeLa S3 cells were purified by chromatography on DNA-agarose and Blue Sepharose. A molecular mass of 112000 Da was found for all three polymerases. Fragmentation of polyacrylamide-gel-embedded polymerases with cyanogen bromide, and subsequent analysis of the fragments by polyacrylamide gradient gel electrophoresis, showed great similarities with regard to fragment sizes. The amino acid composition of the pig thymus enzyme was very similar to that of the polymerase from Ehrlich ascites tumor cells, and the terminal amino group appeared to be blocked. The HeLa polymerase electrofocused in two peaks at pH 8.8 and 5.5, while the Ehrlich ascites tumor cells and the pig thymus enzyme focused in single peaks at pH 9.4 and 9.6 respectively. Removal of residual DNA by treatment with hydroxyapatite abolished these differences in apparent isoelectric points; all three polymerases focused at pH 9.8. No important differences were found with regard to the effect of a number of substances on the synthesis of poly(ADP-ribose). Apparent Michaelis constants for NAD of 41 microM, 48 microM, and 34 microM were found for polymerase from Ehrlich ascites tumor cells, pig thymus, and HeLa S3 cells, respectively. All these results indicate that the three polymerases, which represented the major poly(ADP-ribose) polymerase activity in the organisms investigated, are closely related proteins. PMID- 6281002 TI - Superoxide release by zymosan-stimulated rat Kupffer cells in vitro. AB - Kupffer cells were isolated from pronase-perfused rat livers and were maintained as a monolayer culture in a state of high purity and viability. Immediately after contact with zymosan particles, O2 uptake of the Kupffer cells increased fivefold; about 50% of the net oxygen consumed was accounted for as superoxide released into the medium. Concomitantly, a transient burst of luminol-dependent chemiluminescence, an increased activity of NAD(P)H oxidase and a stimulation of the flow of glucose through the hexose monophosphate shunt were observed. Chemiluminescence and O2- production were almost completely inhibited by superoxide dismutase and iodoacetate. Zymosan-induced chemiluminescence was not inhibited in the presence of the non-penetrating thiol reagents, 5,5'-dithio-bis 2-nitrobenzoate and iodoacetyl-sepharose. Iodoacetate acted on the cytosolic glucose-6-phosphate dehydrogenase rather than on NAD(P)H oxidase of the cell membrane. PMID- 6281004 TI - Effects of chronic stimulation at low frequency on the lipid phase of sarcoplasmic reticulum in rabbit fast-twitch muscle. PMID- 6281005 TI - Physicochemical characterization of detergent-solubilized gamma-aminobutyric acid and benzodiazepine receptor proteins from bovine brain. AB - [3H]Muscimol and [3H]flunitrazepam binding activities have been solubilized from bovine cortex using the ionic detergent sodium deoxycholate. The soluble receptor proteins were shown to bind [3H]muscimol with a dissociation constant, Kd, of 12 nM and a binding capacity (Bmax value) of 1.56 pmol/mg protein; gamma-amino[3H] butyric acid with a Kd of 50 nM and Bmax of 1.55 pmol/mg protein; and [3H]flunitrazepam with a Kd of 8 nM and a Bmax of 0.8 pmol/mg protein. Gel filtration of the soluble receptor proteins showed that the gamma amino[3H]butyric acid and [3H]flunitrazepam binding activities comigrated with a Stokes radius of 6.8 nm. The two binding activities were also found to comigrate after sedimentation in a sucrose density gradient. The hydrodynamic properties of the assumed protein-detergent complexes were determined by gel filtration and sedimentation through gradients of sucrose in H2O or 2H2O. Under the conditions employed, the parameters for both the putative gamma-aminobutyric acid and benzodiazepine receptors were: partial specific volume, 0.73 ml g-1; sedimentation coefficient, 12.5 S; molecular weight, 355000; and frictional ratio 1.46. These observations are consistent with the conclusion that the majority of both binding activities solubilized in deoxycholate reside in a single macromolecular complex. However, Triton X-100 selectively solubilized the benzodiazepine binding activity. This suggests that the two binding activities can be at least partially separated. PMID- 6281006 TI - Local anaesthetics including benzyl alcohol activate the adenylate cyclase in Trypanosoma brucei by a calcium-dependent mechanism. AB - 1. The adenylate cyclase of Trypanosoma brucei is activated by local anaesthetics. 2. Activation of adenylate cyclase by the local anaesthetic, benzyl alcohol, requires Ca2+. 3. The kinetics of the presteady state reveal that the activation step occurs prior to and is separate from the catalytic step. 4. The activation step in the presence of Ca2+ can be stimulated some 200-fold and is highly co-operative with respect to benzyl alcohol concentration. 5. The mechanism by which local anaesthetics activate may be by exposing the Ca2+ binding site of the Ca2+ receptor which is oriented towards the hydrophobic regions of the plasma membrane in the basal state. PMID- 6281007 TI - Effects of streptozotocin-induced diabetes mellitus on the binding and uptake of asialoorosomucoid by isolated hepatocytes from rats. PMID- 6281008 TI - Phosphorylation of 40-S ribosomal subunits by cAMP-dependent, cGMP-dependent and protease-activated protein kinases. AB - The phosphorylation of 40-S ribosomal subunits by cyclic-nucleotide-dependent and protease-activated protein kinases from rabbit reticulocytes was studied in vitro. Under optimal conditions the cAMP-dependent protein kinases incorporated up to 2 mol phosphate/mol S6. The electrophoretic mobility of S6 following phosphorylation indicated that this value was not an average for a population of maximally phosphorylated and non-phosphorylated S6 but represented a uniform population of diphosphorylated 40-S ribosomal subunits. Tryptic digests of S6 were analyzed by two-dimensional fingerprinting following phosphorylation with the cAMP-dependent protein kinase; two phosphopeptides, A and B, were observed. When 40-S ribosomal subunits were examined with the cGMP-dependent protein kinase, 1 mol phosphate was incorporated/mol S6. Upon analysis of the phosphopeptides obtained with the cGMP-dependent protein kinase, only peptide A was observed. S6 was also modified by a cyclic-nucleotide-independent protein kinase, protease-activated kinase II, following activation of the enzyme by limited proteolytic digestion. These findings suggest that a multiple protein kinase system may regulate the phosphorylation state of S6. A second ribosomal protein, S10, was phosphorylated by a different cyclic-nucleotide-independent protein kinase, protease-activated kinase I, and up to 1 mol phosphate was incorporated. PMID- 6281009 TI - The glucose/glucose-6-phosphate cycle in the periportal and perivenous zone of rat liver. AB - Periportal and perivenous hepatocytes contain different activities (V) of antagonistic key enzymes such as glucokinase and glucose-6-phosphatase. In order to get an insight into the metabolism of the periportal and perivenous area the flux rates (v) of the glucose/glucose-6-phosphate cycle were calculated on the basis of the Michaelis-Menten equation using the measured zonal concentrations of glucose and glucose 6-phosphate, the zonal activities of glucokinase and glucose 6-phosphatase previously reported and the half-saturating substrate concentrations (Km) of the two enzymes found in the literature. The concentrations of glucose were obtained as a first approximation by measuring the concentrations in portal (= periportal) and hepatovenous (= perivenous) blood; those of glucose 6-phosphate were calculated from the levels determined in microdissected periportal and perivenous liver tissue. The calculations showed (a) that the overall cycling rates agreed remarkably well with those reported for intact animals and (b) that during a normal feeding rhythm the periportal zone should catalyze net glucose output and the perivenous zone should mediate net glucose uptake, as proposed by the model of 'metabolic zonation'. PMID- 6281010 TI - An extracellular polysaccharide produced by Zoogloea ramigera 115. AB - A weakly acidic polysaccharide was purified from the extracellular zoogloeal matrix produced by Zoogloeal ramigera 115. The purified polysaccharide was homogeneous as judged by sedimentation analysis, and the average molecular weight was estimated to be about 10(5) by gel permeation chromatography of the fully methylated preparation. The polysaccharide was composed of D-glucose, D-galactose and pyruvic acid in an approximate molar ratio 11:3:1.5. On the basis of methylation, periodate oxidation, Smith degradation and partial hydrolysis, the following highly branched structure was deduced for the polysaccharide: a long chain mainly consisting of beta 1 leads to 4-linked glucose residues branching at the C-3 or C-6 position of galactose residues which are present in beta 1 leads to 4 or beta 1 leads to 3 linkages as the minor component of the long chain; pyruvic acid residues, the sole acidic component, are linked to the nonreducing end and/or 1,3-linked glucose residues through 4,6-ketal linkages. The purified polysaccharide was not readily soluble in water and had a high affinity for several metallic ions (e.g, 0.25 mumol Fe3+/mg, and 0.17 mumol Fe2+ mg). Upon addition of metallic ions (1 mM) to a gelatinous aqueous solution of the polysaccharide (K+ form, 0.125%), more than 80% of it immediately coprecipitated out with them. PMID- 6281011 TI - The electrochemical proton gradient of Saccharomyces. The role of potassium. PMID- 6281012 TI - Direct control of glycogen metabolism in the perfused rat liver by the sympathetic innervation. AB - The mode of action of hepatic nerves on the metabolism of carbohydrates was studied in the rat liver perfused in situ. 1. Electrical stimulation of the nerve bundles around the hepatic artery and the portal vein resulted in an increase of glucose and lactate output, an enhancement of phosphorylase a activity and a decrease of portal flow. 2. Sodium nitroprusside prevented the hemodynamic changes after nerve stimulation without affecting the metabolic alterations. 3. Phentolamine or an extracellular calcium level below 300 mumol x 1(-1) abolished both hemodynamic and metabolic changes after nerve stimulation, while propranolol or atropine were without effect. 4. Norepinephrine infusion mimicked nerve stimulation only at the highly unphysiological concentration of 0.1 microM; it was not effective at a concentration of 0.01 microM, which might be reached in the sinusoidal blood due to an overflow from intrahepatic synapses. The present results suggest that, in rat liver, glycogen breakdown is regulated by alpha sympathetic nerves directly rather than indirectly via hemodynamic changes or via norepinephrine overflow. PMID- 6281013 TI - Mitochondrial adenylate kinase from chicken liver. Purification characterization and its cell-free synthesis. AB - Mitochondrial adenylate kinase has been purified 5400-fold from chicken liver extract in an overall yield of 36%. The purified enzyme has a specific activity of 810 U/mg, a molecular weight of 28 000, and the following amino acid composition: 21 aspartic acid or asparagine, 14 threonine, 17 serine, 27 glutamic acid or glutamine, 16 proline, 22 glycine, 22 alanine, 15 valine, 6 methionine, 11 isoleucine, 29 leucine, 5 tyrosine, 7 phenylalanine, 16 lysine, 7 histidine, 19 arginine, 3 half-cystine, and no tryptophan, totalling 257 residues. The purified enzyme has one disulfide bond and one sulfhydryl group. The disulfide bond is related to the active conformation of the enzyme, whereas the sulfhydryl group does not contribute to the enzyme activity. The sulfhydryl group is easily oxidized in the presence of Cu2+ resulting in the formation of dimer with about one half of the specific activity of the monomer. The enzyme is similar to porcine heart mitochondrial adenylate kinase in antigenicity but different from chicken cytosolic adenylate kinase. Mitochondrial adenylate kinase was synthesized in the mRNA-dependent rabbit reticulocyte lysate system programmed with total chicken liver RNA. The mobility in sodium dodecylsulfate gel electrophoresis of the product obtained in vitro was the same as that of the purified mitochondrial adenylate kinase. This evidence indicates that the mitochondrial adenylate kinase is synthesized as a polypeptide with a molecular weight indistinguishable from that of the mature protein. PMID- 6281014 TI - Prognostic significance of paratracheal soft tissue in lung cancer. PMID- 6281015 TI - Relation between cervical carcinoma and HSV-2 infection: herpetic agent in tumoral cells identified by immunofluorescence. PMID- 6281016 TI - Evaluation of renal ischemia with 99mTc-pyrophosphate. AB - Renal ischemia injury was induced by transient 30-min occlusion of the main artery in five rabbits and by transient 60-min occlusion in four rabbits. The vessels to the contralateral kidneys were ligated in all nine animals and in six additional control rabbits. Renal blood flow was restored immediately following the transient ischemia. Twenty-four hours later, 99mTc-pyrophosphate was injected and renal uptake was monitored for 100 s and 10-min intervals for 90 min following injection. At 90 min postinjection, the animals were killed and the percent injected dose remaining in the kidneys was calculated for three animals in each group. At 60 min postinjection, in vivo activity in the 30-min ischemic kidneys was 2.3 times greater than that of controls, and activity in the 60-min ischemic kidneys 4.6 times greater than that of controls. These results suggest that 99mTc-pyrophosphate scanning may be useful in assessing slight ischemic damage to the kidney. PMID- 6281017 TI - Scintigraphic studies in patients with medullary carcinoma of the thyroid. AB - Findings on pre-operative thyroid scintiscans in 29 patients with medullary carcinoma of the thyroid were compared with results of palpation and pathological findings. Six patients had a normal uptake of 131I or 99mTc in tumours 1.5--8 cm in diameter. When compared with 111 pre-operative scintiscans in other thyroid carcinomas, the frequency of normal uptake in medullary carcinomas was high, 19% versus 0--6%. Gallium-67 scintiscans were performed in nine patients. Six patients had extensive tumours, but accumulation of 67Ga appeared in only one patient. PMID- 6281018 TI - Contralateral correction in lateral lung imaging in phantom and clinical studies. PMID- 6281019 TI - The influence of reducing agents on the composition of 99Tc-complexes: implications for 99mTc-radiopharmaceutical preparation. AB - The use of formamidine sulphinic acid as a reducing agent in the presence of technetium-99-pertechnetate and diethyldithiocarbamate ligand has been shown to yield a complex containing a Tc = CO bond. The carbon monoxide present in this complex originates from the reducing agent itself. Evidence is presented which suggests that this carbonyl complex is present in the 99mTc diethyldithiocarbamate preparation obtained by the use of formamidine sulphinic acid as the reducing agent. Use of hydrazine as a reducing agent yields a complex containing a Tc = N bond. It is apparent that when reducing agents such as hydrazine or formamidine sulphinic acid are used in the preparation of 99mTc radiopharmaceuticals, the possibility of the formation of complexes structurally different to those obtained by the use of stannous chloride must be considered. PMID- 6281020 TI - Scintigraphic demonstration of a leiomyosarcoma of the stomach. PMID- 6281022 TI - Iodine-induced hypothyroidism and goiter following lipiodolTM lymphography. AB - Hypothyroid goiter is a rare but well recognized complication following long term administration of iodine containing expectorants and disinfectants in children. Only few reports exist on iodine-induced hypothyroidism after a single injection of the iodized radiopaque dye Lipiodol. A 15-year-old boy with previously normal thyroid function is described who developed hypothyroid goiter within six weeks following bipedal lymphography. Urinary iodine excretion was extremely elevated up to 18 mg/day while serum concentrations of total thyroxine were below the euthyroid range and thyrotropin levels were elevated. After oral L-thyroxine treatment the goiter disappeared. Thyroid function remained normal when treatment was discontinued after five months although iodine excretion was still 50 times higher (2.5 mg/day) than in normal age matched children. The observed alterations of the thyroid gland were caused by a long lasting Wolff-Chaikoff effect with a delayed adaptation to high iodide concentrations. PMID- 6281021 TI - Immunological and virological investigations in Down's syndrome. AB - Lymphocyte responsiveness to phytohaemagglutinin and viral antigens was studied in children with Down's syndrome and in controls. Mitogen-responsiveness in the patients was significantly reduced as compared to the control values. Using the lymphocyte transformation test, trisomic patients showed more than a twofold increase in sensitivity to herpes simplex virus as compared to controls. The same test did not show any essential difference between the two groups when adeno- and influenza viruses were used. Immunofluorescence technique, with specifically conjugated antiviral sera, permitted the detection of specific fluorescence in 30% of the patients with Down's syndrome indicating the presence of oncogenic adenovirus type 12 antigen in the circulating lymphocytes. No antibodies--or only very low titres--against adeno- and herpes simplex viruses were demonstrated in the sera of trisomic patients. Mononuclear leukocytes from these patients often showed structural alterations. The incidence of infectious herpes simplex virus and Candida albicans in the saliva of patients was higher than in the control group. It seems that Down's syndrome involves partial disturbance of both the cellular and humoral immune functions--at least with respect to certain viral antigens. PMID- 6281023 TI - Gonadotropin-releasing hormone analogues for prostate cancer: untoward side effects of high-dose regimens acquire a therapeutical dimension. AB - Gonadotropin-releasing hormone (Gn-RH), a hypothalamic decapeptide, mediates the synthesis and release of pituitary luteinizing hormone (LH). Synthetic Gn-RH analogues with prolonged duration of action are therapeutically administered with 'pro-fertility' intention to overcome endogenous Gn-RH deficiency, i.e. hypogonadotropic hypogonadism and related disorders. When Gn-RH analogues are chronically administered in high supraphysiological dosages, however, a paradoxical effect of pituitary overstimulation becomes evident: gonadotropin release is reduced, Leydig cell responsiveness is impaired, and testosterone secretion is blocked to castrate levels. Metabolic mechanisms involved include desensitization of the pituitary to Gn-RH, down-regulation of Gn-RH receptors, the depletion of the releasable LH pool, the breakdown of physiological feedback mechanisms, as well as direct interactions of the Gn-RH analogue with the Leydig cell. It is tempting to assume that, on the basis of this untoward side effect, high dose Gn-RH analogues will acquire a therapeutical dimension in the palliative treatment of prostate cancer. The therapeutical effect of castrate levels of testosterone is achieved without orchiectomy, cardiovascular side effects of estrogen are avoided, and the drug is conveniently applicable via the pernasal route. From the data available today Gn-RH analogues in overstimulatory dosage can be expected to be safe and effective in the palliative treatment of prostate cancer and would thus prove a true alternative to conventional contrasexual measures. PMID- 6281024 TI - Necessary extent of lymph node dissection in testicular tumours. A histopathological investigation. AB - The findings of a prospective topographic histopathological study of the lymphatic spread of testicular tumours are reported. The study includes 150 patients, 9 of whom were subjected to exploratory laparotomy while 141 patients underwent retroperitoneal lymph node dissection. Lymphatic spread occurs in accordance with a quite definite pattern, atypical metastases were observed in only 3 of the 150 patients. The material was analyzed with respect to the possibilities of using limited unilateral dissections. The results show that by observing appropriate safety precautions, including frozen section histology, limited unilateral dissections can be used more frequently. PMID- 6281025 TI - The expression and functional involvement of nuclease- specific idiotype on nuclease-primed helper T cells. AB - The expression and functional significance of idiotypic determinants on antigen specific helper T (Th) cell populations for responses to Staphylococcal nuclease (Nase) were evaluated in an in vitro antibody response system. Trinitrophenyl (TNP)- specific plaque-forming cell responses to TNP-conjugates of Nase (TNP Nase) were shown to require the cooperation of Nase-primed Th cells as well as unprimed B and accessory cells. The expression on these antigen-primed Th cells of idiotypic determinants cross-reactive with those on anti-Nase antibodies was demonstrated by the specific elimination of Th cells for TNP-Nase by treatment with affinity-purified anti-idiotypic antibodies plus complement. The susceptibility of Nase-primed Th cells to elimination by such treatment was specific in that anti-idiotypic antibodies affected Th cells only from strains normally expressing the same (or a cross-reactive) idiotype on anti-Nase antibodies. A functional role of the idiotypes expressed on Nase-primed Th cells was suggested by the fact that anti-idiotypic antibody present throughout the period of culture, in the absence of complement, suppressed responses to TNP-Nase in an antigen- and strain-specific manner. It was further shown, by cell mixing experiments, that this inhibition appeared to occur at the level of the Th cells and was not dependent on the strain of origin of the B cells. Thus, antigen specific Nase-primed Th cells express strain-specific idiotypic determinants cross-reactive with, or identical to, those of anti-Nase antibodies. These cell surface idiotypic determinants appear to be functionally involved in the activity of Th cells for the induction of antibody responses to TNP-Nase in vitro. PMID- 6281026 TI - Human monoclonal autoimmune antibody produced in vitro: rheumatoid factor generated by Epstein-Barr virus-transformed cell line. AB - The infection of selected lymphocytes from a rheumatoid arthritis patient with Epstein-Barr virus resulted in an immortalized cell line that secretes a monoclonal rheumatoid factor (RF). The cloned line has been growing for more than 24 months, and constantly produces a monoclonal IgM, lambda, 19S, RF (1-2 micrograms/ml/106 cells). The RF agglutinates human and rabbit IgG (but not IgM) and also protein A-coated erythrocytes, but fails to do so to mouse, goat and swine IgG-coated erythrocytes. When bound to immune complexes, this monoclonal RF dose not bind complement. In the cell supernatant RF is the only immunoglobulin and it comprises approximately 5% of the total proteins. The affinity of RF to aggregated human IgG, as detected in inhibition experiments, is higher than that of Fc receptors found on human non-T lymphocytes, K562 and Daudi cell lines. PMID- 6281027 TI - Involvement of alpha 1- and alpha 2-adrenoceptors in the vasoconstriction caused by ergometrine. AB - In pithed normotensive rats, the pressor effects evoked by i.v. serotonin (5-HT) and ergometrine were analysed using the relatively selective alpha 1-, alpha 2- and serotonin antagonists prazosin, yohimbine methysergide, respectively. The pressor response to ergometrine was reduced by both prazosin and yohimbine but only moderately affected by methysergide. On the other hand, the rise in diastolic pressure brought about by serotonin was strongly depressed by methysergide but not influenced by the blockade of either alpha 1- or alpha 2 adrenoceptors. The calcium antagonist nifedipine shifted the dose-pressor response curve of ergometrine to the right in a dose-related, non-parallel manner. The maximum pressor response was diminished by nifedipine. In contrast, the rise in pressure provoked by serotonin was not affected by nifedipine. The results thus show that in the pithed rat the vasoconstrictor response to ergometrine is mediated by alpha-adrenoceptors (both alpha 1 and alpha 2) rather than by serotonin receptors. These findings question the role of serotonin receptors in the vasoconstriction induced by ergometrine. PMID- 6281028 TI - Alpha 1- and alpha 2-adrenoceptors in rat cerebral cortex: effects of neonatal treatment with 6-hydroxydopamine. AB - Neonatal 6-hydroxydopamine treatment was used to destroy the noradrenergic nerve terminals in rat cerebral cortex and thus give some insight into the in vivo regulation of alpha-adrenoceptor subtypes, which in turn provides information concerning the anatomical localization of alpha 1- and alpha 2-adrenoceptors. Treatment of rats in the neonatal period with 6-OHDA causes an irreversible decrease in noradrenaline levels of the cerebral cortex compared to controls. Differences in [3H]clonidine and [3H]prazosin binding in the cerebral cortex occurred which varied depending upon the time elapsed between denervation and the binding assay. In rats aged 7-14 days there was a 20% decrease in the number of alpha 2-adrenoceptors and a slight increase in alpha 1-adrenoceptors. In older rats (45-50 day old) both types of alpha adrenoceptors were increased. Results of this study indicate that alpha 2-adrenoceptors located on presynaptic noradrenergic terminals represent only a small proportion of the total alpha 2 adrenoceptors in rat cerebral cortex. Increases in the binding capacity after 67 OHDA treatment indicate an up-regulation phenomenon affecting alpha 1 adrenoceptors and alpha 2-adrenoceptors located on structures other than noradrenergic nerve endings. PMID- 6281029 TI - Beta-receptors during aging in respiratory tissues. AB - Specific [125I] hydroxybenzylpindolol binding (33 fmol/mg protein) was detected in tracheal tissues from middle aged (417 g) and old (757 g) guinea pigs but binding was not measurable in tracheal tissues from young (118 g) animals. Similarly, receptor density increased in bronchial and parenchymal tissues during development but receptor affinity did not change. For any age, the receptor densities were parenchyma greater than bronchi greater than trachea (20/7/1); receptor affinities were identical. The potency of l-isoproterenol in relaxing bronchial muscle was reduced during development. In vivo, salbutamol reduced airway reactivity to histamine and was most potent in animals exhibiting airway hyperreactivity. In addition, l-propranolol sensitized airway muscle to the bronchoconstrictor effects of histamine in young guinea pigs; in old guinea pigs of the same airway reactivity, l-propranolol did not affect the induced bronchoconstriction. Our data suggest that there is a reduced sensitivity of airway muscle to catecholamines during development which may be due, in part, to increased density of beta-adrenoceptors which are not involved in eliciting the physiological response. PMID- 6281030 TI - Pulmonary opiate receptor activation evokes a cardiorespiratory reflex. AB - The administration of [D-Ala2,Met5]enkephalinamide (D-AME) and [D Ala2,Leu5]enkephalinamide (D-ALE) into the right atrium of decerebrate rats caused bradycardia, a slight transient biphasic blood pressure response and apnea within 1-2 sec. Apnea was followed by rapid shallow breathing. These effects were dose related (1-1000 micrograms/kg) and blocked by pretreatment with naloxone. Atropine blocked the bradycardia. Sectioning the vagi at the level of the diaphragm did not affect the responses, whereas bivagotomy below the cardiac branches abolished all responses. The triad of responses was attributed to a reflex action arising from vagal afferents within the lung. These results were confirmed in paralyzed , artificially ventilated animals. In these animals, the enkephalin analogues produced a cessation of phrenic nerve (PN) activity followed by a decrease in the duration of bursts. The recurrent laryngeal nerve (RLN) was concomitantly excited in a continuous decremental fashion. This excitation was independent of PN inhibition. Recordings of single and near single pulmonary vagal afferents demonstrated no effect of D-AME or D-ALE on stretch and irritant receptors. However, type J-receptors were stimulated. PMID- 6281031 TI - Alterations in local cerebral glucose utilization during chronic treatment with an ACTH 4-9 analog. AB - Adrenocorticotrophic hormone (ACTH) and structural analogs of the fragment ACTH4 9 have marked effects upon behavior (particularly in relation to memory processes). The effects of the chronic (10 day) administration of the potent analog (Met(O2)4,D-Lys8,Phe9]ACTH4-9(100 microgram/kg per day i.p,) upon local cerebral glucose utilization have been examined using the autoradiographic 2 deoxyglucose technique in the attempt to identify the neuroanatomical sites involved functionally in the central actions of this ACTH4-9 analog. Of the 49 anatomically discrete regions examined, significant increases in glucose utilization were observed only in the hippocampus (stratum molecular lacunosum lacunosum and parasubiculum, increased by 16 and 17% respectively), the anterior nucleus of the thalamus (by 23%) and anterior cingulate cortex (by 30%). The highly localized alterations in glucose utilization which were observed following treatment with this ACTH4-9 analog provide evidence for the functional involvement of a hippocampal-anterior-thalamic-anterior cingulate cortical circuit in the actions of this peptide fragment. PMID- 6281032 TI - An isolated perfused dog lung preparation for the study of cyclic GMP metabolism: effects of sodium nitroprusside and oxygen. AB - The intact, isolated perfused dog lung was evaluated as a model for studies directed at defining the role of oxidative modulation of lung cyclic GMP metabolism in pulmonary function. Sodium nitroprusside added to the perfusion blood increased the cyclic GMP content of lung over 4-fold in a dose-dependent manner. Although sodium nitroprusside administration caused changes in lung vascular resistance, these occurred independently of the changes in cyclic GMP. Ventilation of lungs with a high oxygen gas mixture containing 95% O2. 5% CO2 acutely increased the cyclic GMP content of lungs after 15 min from 1.3 +/- 0.06 (mean +/- SE) to 3.4 +/- 0.12 pmol cyclic GMP/mg protein. Cyclic GMP levels returned toward control during continued ventilation with the high oxygen concentration. The oxygen-induced elevation of lung cyclic GMP content was not accompanied by changes in lung vascular resistance. The results indicate that the isolated perfused lung may be useful in studies of cyclic GMP, tissue oxidation and pulmonary function. PMID- 6281033 TI - Cholinergic inhibition of acetylcholine release in the electric organ of Torpedo. AB - The possibility that acetylcholine (ACh) may inhibit its own release from nerve terminals by acting on presynaptic receptors has been investigated using the electric organ of Torpedo marmorata. ACh release was analysed by electrophysiological and biochemical methods conjointly. Oxotremorine, at micromolar concentrations, depressed nerve-electroplaque transmission by reducing the amount of ACh released by nerve impulses. This effect was competitively antagonised by nanomolar concentrations of atropine or methylatropine. Other muscarinic agonists, betanechol, pilocarpine and muscarine, however, failed to depress transmission but choline was effective at high concentrations. Anticholinesterase drugs, physostigmine, neostigmine or fluostigmine (diisopropylfluorophosphate, given as pretreatment and subsequently washed out) markedly depressed evoked ACh release. When cholinesterase was inhibited, the addition of oxotremorine or exogenous ACh caused a further depression of ACh release. Atropine was found to be very effective in reversing the depression of transmitter release produced by anticholinesterases. Looking for the mechanism of these presynaptic changes, we found that oxotremorine had little, if any, effect on the size of the ACh store of the tissue, on the compartmentation of ACh inside and outside synaptic vesicles, or on the rate of ACh turnover. The changes induced by oxotremorine cannot be explained by a reduction in calcium entry, since the presence of oxotremorine did not change the uptake of 45Ca observed after repetitive stimulation. Electrophysiological techniques were used to test for an effect of atropine in experiments where transmission of one impulse was expected to depress ACh release by subsequent impulses. This depression was not affected atropine, making it unlikely that the 'muscarinic' inhibition of ACh release has a role as a short-term feedback regulation of transmission. A second possibility is that oxotremorine (and external non-hydrolysed ACh) can enter the presynaptic membrane and interfere with the mechanism of transmitter release. PMID- 6281034 TI - Temporary spinalization reverses the inhibitory effect of cyclazocine on the nociceptive response of rabbit spinal dorsal horn lamina V-type neurons. AB - Cyclazocine (0.5 mg/kg i.v.) inhibited the activity of lamina V-type neurons of the rabbit spinal dorsal horn induced by intra-arterial injection of bradykinin. This inhibition was reversed during temporary spinalization by cold block. Thus, cyclazocine probably depresses nociceptive transmission in the spinal dorsal horn by facilitating descending inhibitory systems which originate from supraspinal structures. PMID- 6281035 TI - Modulation of benzodiazepine receptor binding: insight into pharmacological efficacy. AB - The effects of GABA on the binding of analogues of benzodiazepines, triazolopyridazines, beta-carbolines and imidazodiazepines were examined in ligand/[3H] flunitrazepam competition experiments. GABA increased the potency of anxiolytics, like flunitrazepam, whereas the potency of benzodiazepine antagonists, like Ro15-1788, was largely insensitive to the influence of GABA. Several other agents including pyrazolopyridines, barbiturates and etomidate caused a chloride dependent enhancement of [3H] flunitrazepam binding but not an enhancement of [3H] propyl-beta-carboline-3-carboxylate binding. PMID- 6281036 TI - SA446, a new orally active converting enzyme inhibitor: antihypertensive action and comparison with captopril in stroke-prone spontaneously hypersensitive rats. AB - The converting enzyme inhibitors (CEI) SA446 and Captopril (CAP) were given orally at a dose of 50 mg/kg per day to adult stroke-prone spontaneously hypertensive rats (SHRSP) over a period of four weeks. Both CEI lowered arterial blood pressure (BP) to a similar extent. CAP was more inhibitory on the plasma renin-angiotensin system (RAS) than SA446. Both CEI lowered urinary aldosterone excretion but had little (SA446) or no (CAP) natriuretic effect. CAP reduced the pressor responses to intravenous (i.v.) angiotensin I (ANG I) more (52%) than SA446 (18%) and potentiated the depressor responses to i.v. bradykinin more (fortyfold) than SA446 (tenfold). In contrast, SA446 treatment reduced the pressor responses to intracerebroventricular (i.c.v.) ANG I by 21% and led to a rise in the hypothalamic renin concentration. Oral CAP treatment for four weeks did not produce these signs of a brain converting enzyme inhibition. It is concluded that SA446 is equally as antihypertensive as CAP in SHRSP. SA446 appears to penetrate more readily into the brain and to exert its action partly through inhibition of the brain RAS which is known to be stimulated in SHRSP. PMID- 6281037 TI - Prostaglandin F2 alpha modifies the action of norepinephrine on alpha- and beta adrenoceptors of isolated rat uterus. PMID- 6281038 TI - Chronic propranolol treatment decreases cardiac beta-adrenoceptors in spontaneously hypertensive rats. AB - Spontaneously hypertensive (SHR) rats were treated for 6 months with 41.6 or 94.6 mg/kg per day of (+/-)-propranolol. Compared to untreated SHR rats, the heart rate and blood pressure in both treated groups were decreased. The cardiac beta adrenoceptor concentration was similar in the untreated and low dose (+/-) propranolol-treated group but was decreased by 30% after treatment with the high (+/-)-propranolol dose. The results suggest that high dose (+/-)-propranolol treatment reduces the cardiac beta-adrenoceptor concentration. PMID- 6281039 TI - Benzomorphan binding sites in rat lumbo-sacral spinal cord. AB - The rat lumbo-sacral spinal cord contains a homogeneous population of opiate binding sites labelled with high affinity (KD = 0.21 +/- 0.04 nM) by [3H]etorphine and lower affinity (KD = 2.2 +/- 0.4 nM) by [3H]ethylketocyclazocine. Benzomorphan drugs are potent competitors for these binding sites while morphine and enkephalin display a low affinity. These binding sites have binding properties which are distinct from the mu-, delta-, and also kappa-sites but are very similar to those of the benzomorphan sites characterized in rat brain. PMID- 6281040 TI - Studies on viral respiratory disease in laboratory cats. I. Isolation of feline herpesvirus and choice of proper disinfectant. AB - In the laboratory cat colony consisted of 14 Korats and 8 Japanese native cats, the disease characterized by upper respiratory signs occurred. Seven cytopathogenic agents were isolated from the oropharyngeal swabs and necropsy materials of affected cats by means of inoculation to primary feline kidney cell cultures. One of the isolates (KS-1 strain) was identical with feline herpesvirus (FHV), and appeared to be a causal agent of the trouble. Properties of the KS-1 strain were in general agreement with the known strains of FHV. Serologic examination and clinical analysis suggested that Japanese native cats became carriers, re-excreted the virus and spread it to susceptible cats. Virucidal activities of several disinfectants against FHV, comparing with a feline calicivirus (FCV), were studied and the following conclusions reached: 1) Sodium hypochlorite, iodine complex, benzethonium chloride and chlorhexidine were effective against FHV at commonly used concentrations. 2) Among chemicals tested sodium hypochlorite was most effective against FCV and followed by iodine complex. 3) Benzethonium chloride and chlorhexidine had little efficacy against FCV at commonly used doses. PMID- 6281041 TI - [Elimination of mouse hepatitis virus infection in human malignant tumors transplanted into athymic (nude) mice (author's transl)]. AB - Mouse hepatitis virus (MHV) infection occurred in athymic (nude) mice transplanted with human malignant tumors and 28 out of 38 tumor lines were contaminated with MHV. These 28 tumor lines were passaged in pathogen-free nude mice and 15 of these tumor lines (53.6%) became MHV-free. The MHV infection was eliminated in more than half of the tumor lines by passaging. PMID- 6281042 TI - [Spontaneous liver neoplasms in Sprague-Dawley rats]. PMID- 6281043 TI - Cell growth and tRNA-lys4 synthesis in mouse 3T3 cells. PMID- 6281044 TI - Stability of parental mitochondrial DNA species and expression of nuclear ribosomal RNA genes in mouse-rat hybrid cells. PMID- 6281045 TI - Genetic complementation in hybrid cells derived from two metabolic co-operative defective mammalian cell lines. PMID- 6281046 TI - Correlations between prestalk-prespore tendencies and cAMP-related activities in Dictyostelium discoideum. PMID- 6281047 TI - Calcium and ATP regulation of the oscillatory torsional movement in a triton model of Physarum plasmodial strands. PMID- 6281048 TI - Loss of transferrin receptors following induced differentiation of HL-60 promyelocytic leukemia cells. PMID- 6281049 TI - Stimulation of liver cell DNA synthesis by oncomodulin, an MW 11 500 calcium binding protein from hepatoma. PMID- 6281050 TI - Correlation of increased nuclease activity with enhanced virus reactivation. PMID- 6281051 TI - Enhancement in 'apparent' membrane microviscosity during differentiation of embryonal carcinoma cells induced by retinoids. PMID- 6281052 TI - Stereospecific [3H]naloxone binding associated with opiate receptors in bovine retina. PMID- 6281053 TI - Organophosphate metabolites of the human and rabbit crystalline lens: a phosphorus-31 nuclear magnetic resonance spectroscopic analysis. PMID- 6281054 TI - Abrogation of infectivity of mouse mammary tumor virus by reserpine. PMID- 6281057 TI - Cytomegalovirus-like particles in the red-backed vole submandibular gland. PMID- 6281056 TI - An increase in Sendai virus-induced cell fusion of erythrocytes infected with Plasmodium chabaudi. AB - The extent of cell fusion induced by Sendai virus was examined in erythrocytes infected with Plasmodium chabaudi. An increase in cell fusion of erythrocytes with Ehrlich tumor cells and of erythrocytes with erythrocytes was observed with the infected erythrocytes. However, agglutination by the virus was not changed between erythrocytes of normal and malarial mice. These results indicate that the increase in cell fusion occurred in the process of membrane fusion, suggesting that some membrane property of Plasmodium-parasitized erythrocytes is changed in terms of Sendai virus-induced cell fusion. PMID- 6281058 TI - Electrically induced fluorescence as a method for studying benzo[a]pyrene binding to DNA. PMID- 6281055 TI - Organization and expression of the poxvirus genome. AB - Poxviruses comprise a large group of very complex animal DNA viruses which replicate in the cytoplasm of infected cells. Vaccinia virus, the most studied poxvirus, has a linear, double stranded DNA genome with an approximate molecular weight of 120 x 10(6) (180 kilobase pairs). The two strands of the DNA molecule are naturally cross-linked at both termini. In addition, the vaccinia virus genome contains very long inverted terminal repetitions of approximately 10 kilobase pairs which are further characterized by the presence of direct tandem repeats of a 70-base-pair sequence arranged in two blocks of 13 and 17 copies, respectively. A central region of the genome is highly conserved between different orthopoxviruses. In contrast, the ends are hypervariable and may contain extensive deletions and complex, symmetrical sequences rearrangements. Vaccinia virus gene expression is divided into two stages. Early in infection, RNA complementary to one half of one strand-equivalent of the genome is transcribed within subviral particles by the virion-associated RNA polymerase. Later in infection, after DNA replication, RNA complementary to one entire strand equivalent is transcribed. RNA made late in infection is very heterogeneous in length and a large fraction of it contains self-complementary sequences. Late genes are clustered near the central region of the genome. Vaccinia virus mRNAs do not appear to be synthesized by a splicing mechanism. PMID- 6281059 TI - Digestion of chromatin to H1-depleted 166 basepair particles by Ca2+/Mg2+ dependent endonuclease. PMID- 6281060 TI - Coincidence of subnuclear distribution of poly(ADP-ribose) synthetase and DNA polymerase beta in nuclei of normal and regenerating liver. PMID- 6281061 TI - The effect of digestion with phospholipase C on intrinsic protein phosphorylation in synaptic plasma membrane fragments. PMID- 6281063 TI - Ca2+--Calmodulin dependent myosin light-chain phosphorylating activity in insulin secreting tissues. PMID- 6281064 TI - CD properties of the fast- and slow-folding forms of unfolded ribonuclease A. PMID- 6281062 TI - Simultaneous stimulation of uric acid synthesis and gluconeogenesis in chicken hepatocytes by alpha-adrenergic action of epinephrine and calcium. PMID- 6281065 TI - Changes in phospholipid polar head group turnover in SV40-transformed hamster fibroblasts. PMID- 6281067 TI - o-Type cytochrome oxidase in the membrane of aerobically grown Pseudomonas aeruginosa. PMID- 6281066 TI - Influence of the surface potential on the Michaelis constant of membrane-bound enzymes: effect of membrane solubilization. PMID- 6281068 TI - The structural requirements of substrates of cyclic AMP-dependent protein kinase. PMID- 6281069 TI - Leukotrienes A4 and B4 stimulate the formation of cyclic AMP in human leukocytes. PMID- 6281070 TI - Concerted phosphorylation of endogenous tracheal smooth muscle membrane proteins by Ca2+ . calmodulin-, cyclic GMP- and cyclic AMP-dependent protein kinases. PMID- 6281071 TI - Na+, K+-ATPase of the canine mesenteric artery. AB - Na+,K+-ATPase, the enzymatic moiety that operates as the electrogenic sodium potassium pump of the cell plasma membrane, is inhibited by cardiac glycosides, and this specific interaction of a drug with an enzyme has been considered to be responsible for digitalis-induced vascular smooth muscle contraction. Although studies aimed at localization, isolation, and measurement of the Na+,K+-ATPase activity (or Na+, K- pump activity) indicate its presence in vascular smooth muscle sarcolemma, its characterization as the putative vasopressor receptor site for cardiac glycosides has depended on pharmacological studies of vascular response in vivo and on isolated artery contractile responses in vitro. More recently, radioligand-binding studies using [3H]ouabain have aided in the characterization of drug-enzyme interaction. Such studies indicate that in canine superior mesenteric artery (SMA), Na+,K+-ATPase is the only specific site of interaction of ouabain with resultant inhibition of the enzyme. The characteristics of [3H]ouabain binding to this site are similar to those of purified or partially purified Na+,K+-ATPase of other tissues, which suggests that if Na+,K+-ATPase inhibition is causally related to digitalis-mediated effects on vascular smooth muscle contraction, then therapeutic concentrations of cardiac glycosides could act to cause SMA vasoconstriction. The additional finding from radioligand-binding studies that Na+,K+-ATPase exists in much smaller quantities (density of sites per cell) in SMA than in either heart or kidney may have implications concerning its physiological, biochemical or pharmacological role in modulating vascular muscle tone. PMID- 6281072 TI - Diffusion in lung gas and across alveolar membrane in mammalian lungs. AB - The relative importance of stratified inhomogeneity in the mammalian lung has been considered both theoretically and experimentally. Calculations show that alveolar-capillary diffusion resistance is not limiting for inert gas exchange. Previous model calculations have concluded that stratified inhomogeneity is not likely to be an important mechanism in limiting gas exchange. Experimental data with inspired boluses of inert gas show the presence of stratified inhomogeneity. However, another possible mechanism, related to interaction of diffusion and convection, remains as a possible explanation for these findings. Stratified inhomogeneity appears to play a role in gas exchange in the normal rat, but the role of stratified inhomogeneity in humans and other animals is yet to be determined. PMID- 6281073 TI - Diffusion in gas exchange of insects. AB - The air-filled tracheal system constitutes the organ for gas exchange in terrestrial insects-its finest branches, the tracheoles, contacting individual cells. In the pupal stage, in which the animal lacks significant ventilatory movement, diffusion in the gas phase of the tracheal system constitutes the only mechanism for gas transfer between the environment and the tissues, transport in the hemolymph being insignificant. We have attempted to identify the main sites of diffusional resistance in the tracheal gas system by measuring the evolution of inert gases of low solubility from the pupa of the giant silkworm moth (Hyalophora cecropia). The results are compatible wih a single model in which the resistance to diffusional gas transfer in the tracheal system is concentrated at its opening at the body surface (spiracle). PMID- 6281074 TI - Acquisition of conditioned facial reflexes in the cat: cortical control of different facial movements. AB - The motor cortex plays a role in determining which of three different facial movements is acquired in Pavlovian conditioning experiments. Three separate facial reflexes can be distinguished by recording electromyographic activity from the orbicularis oculi (eye blink) and levator orii (nose twitch) muscles. One in a pure eye blink; a second is a nose twitch; the third is a compound eye blink and nose twitch. Which of these movements is elicited by a click (conditioned stimulus) following associative conditioning is reflected by the pattern of unit activity elicited by the click at the motor cortex. Activity is enhanced, after conditioning, in those units that project polysynaptically to the specific muscles performing the learned movement. This enhancement of activity is, in turn, relatable to an enhanced electrical excitability of the involved neurons. Analogous changes in the excitability of neurons of the motor cortex to applied currents can be produced by local application of cholinergic agents. Iontophoresis of acetylcholine, aceclidine (a cholinomimetic drug), or intracellularly applied cyclic GMP produces changes in single neuron membrane resistance that increase neuronal excitability. The units of the motor cortex that respond preferentially to these agents and to the click conditioned stimuli with short latencies have been identified as pyramidal cells of layer V. The axons of these neurons form the pyramidal tract, a pathway characterized as serving voluntary movement. It appears that this system supports rapid transmission and processing of auditory-motor information used to perform learned movements adaptively, selectively, and discriminatively. PMID- 6281075 TI - Recent approaches to secretion and stimulus-secretion coupling. PMID- 6281076 TI - Properties of calcium receptors that initiate depolarization-secretion coupling. AB - The relationship between the binding of divalent metal (Me) activators Ca and Sr and the secretion of acetylcholine (ACh) was studied quantitatively at frog motor nerve terminals using conventional electrophysiological methods. Experiments were designed to evaluate the assumption that maximal secretion requires occupancy of all receptors by testing for the presence of spare Ca receptors on nerve endings. Such a receptor reserve for Ca would invalidate the simple mass action approach to ACh secretion. Experimental log [Me]-ACh secretion curves constructed to saturation for Ca Sr were consistent with the presence of spare Ca receptors. La3+ (greater than or equal to 0.5 microM) and 2-chloroadenosine (25 microM) were employed as irreversible antagonists of depolarization-secretion coupling. Despite the irreversible occlusion of a proportion of Me receptors increases in the extracellular [Ca] overcame this antagonism while increases in [Sr] did not. These results suggest that Ca can produce maximal ACh release while leaving a proportion of receptors unoccupied or spare. Further support for this contention is provided by the excellent agreement between the values of the equilibrium affinity constant for Sr calculated by methods that do or do not require the assumption of spare receptors. The equilibrium affinity constant for Ca and the efficacies (efficacy reflects the ability of the Me species once bound to evoke ACh secretion) for both Ca and Sr were determined experimentally by using the mathematical framework of receptor theory. These constants were then employed to generate theoretical curves of log [Me]-ACh secretion. The theoretical relationships were similar to the experimental results, which suggests that the motor nerve endings behaves as a pharmacological receptor for Me agonists and antagonists. It is speculated that spare Ca receptors are equivalent to spare Ca channels and the efficacy may reflect the affinity of Me for an intraterminal site associated with ACh release. PMID- 6281077 TI - Calcium-phospholipid interactions in secretory cells: a new perspective on stimulus-secretion coupling. AB - Recent evidence suggests that stimuli delivered to the cell surface may alter Ca availability by promotion membrane phospholipid turnover. Due to the importance of Ca in stimulus-secretion coupling, the study of membrane phospholipid turnover could provide valuable information on the cellular mechanisms of evoked secretion. In the adrenal cortex, Ca is an obligatory requirement for adrenocorticotropin (ACTH)-induced steroid production and release, and recent findings revealed the presence of a Ca-dependent phospholipase A2 localized to the surface of isolated feline adrenocortical cells. This phospholipase is activated by ACTH to promote the release of arachidonic acid from phospholipids and thereby stimulate prostaglandin formation. Further investigations into the fate of radiolabeled arachidonic acid described a rapid and specific turnover of arachidonic acid within a phosphatidylinositol (PI) pool that is independent of changes in de novo synthesis and is characterized as a Ca-dependent hydrolysis of PI, followed by a rapid, selective reacylation of lysoPI. A similar deacylation reacylation reaction involving arachidonyl PI is also found in the rabbit neutrophil when challenged with the formyl-methionyl peptide, F-Met-Leu-Phe, or the Ca-selective ionophore A23187. The relevance of this reaction to secretory phenomena is indicated by its requirement for Ca, its rapid onset, and dose response curves that paralleled those of the secretory response. The Ca-dependent activation of arachidonyl PI turnover triggered by the activation of membranous phospholipase A2 occupies a critical position in the train of events associated with the activation of the secretory process, and one or another of the products of this reaction-prostaglandins, arachidonic acid, and/or lysophospholipids-may participate in the cellular processes that accompany the discharge of secretory product. PMID- 6281078 TI - Secretion of oxygen intermediates: role in effector functions of activated macrophages. AB - The ability of macrophages to secrete reactive oxygen intermediates, such as superoxide or hydrogen peroxide, correlates closely with their capacity to kill trypanosoma, toxoplasma, leishmania, and candida. In this sense, secretion of oxygen intermediates is a biochemical marker of macrophage activation. The close correlation between oxidative metabolism and antimicrobial activity appears to stem from the direct involvement of oxygen intermediates in the killing of the same parasites by the macrophages. Similarly, there seem to be at least three experimental settings in which oxygen intermediates play a major role in nonphagocytic lysis of tumor cells by macrophages: in the presence of phorbol myristate acetate, of antitumor antibody, or of a peroxidase derived from eosinophils. These findings direct attention to antioxidant defenses in tumor cells and parasites. The oxidation-reduction cycle of glutathione is one major pathway used by tumor cells to limit oxidative injury by macrophages and granulocytes. Thus, cytotoxicity is augmented by inhibition of glutathione reductase or glutathione peroxidase, by interruption of glutathione synthesis, or by diversion of glutathione into another pathway. On the other hand, catalase appears to play a prominent role in limiting macrophage effector function against toxoplasma. PMID- 6281079 TI - Calcium and tension regulation in skinned skeletal muscle fibers. AB - This article reviews the skinned muscle fiber techniques and approaches that have been used in our laboratory in the study of both tension and Ca regulation over the past 15 years. The impetus for using these fibers was the finding that most membrane Cl channels in crayfish muscle are located within the transverse tubules at the junction (diad) with the sarcoplasmic reticulum (SR). This observation led to the postulate that a change in Cl concentration around the SR would cause it to release stored CA. This could be tested by using skinned fibers. Tensions induced by Cl release of SR- Ca or by direct application of Ca are highly dependent on the concentration of substrate (MgATP). A substrate-inhibition formulation accurately summarizes this dependence, but it is not integrated with current biochemical schemes of ATP hydrolysis. To facilitate development of a broad model for substrate regulation of tension, which includes concepts of a cross-bridge cycle, we developed a mammalian skinned fiber preparation and techniques for monitoring regulation in single fibers. Both optical and mechanical techniques are used to investigate contractile protein interactions and Ca binding. PMID- 6281080 TI - Calmodulin: an overview. AB - Calmodulin is a 16,700-dalton Ca2+-binding protein ubiquitous in the eukaryotes. It has no intrinsic enzymatic activity, but it regulates a wide spectrum of enzymes that control many basic cellular processes, ranging from the metabolism of cyclic nucleotides, Ca2+, and glycogen to contractile activity and stimulus secretion coupling. Mounting evidence now indicates that calmodulin is the major intracellular Ca2+ receptor that remained elusive despite three decades of extensive work by many investigators. PMID- 6281081 TI - Insulin release and protein phosphorylation: possible role of calmodulin. AB - Both Ca2+ and cyclic AMP (cAMP) are implicated in the regulation of insulin release in the pancreatic beta cell. In hamster insulinoma cells used in our laboratory to study the mechanism of insulin release, Ca2+ and cAMP trigger secretion independently. Concomitant with stimulation of the secretory apparatus both cAMP and Ca2+ promote phosphorylation of distinct insulinoma cell proteins. Calmodulin may be involved in the stimulation of insulin release and protein phosphorylation induced by Ca2+ influx. The Ca2+-dependent protein kinase of the insulinoma cell is activated by exogenous calmodulin and blocked by trifluoperazine, and inhibitor of calmodulin action. This drug also inhibits glucose-induced insulin release in pancreatic islets. In insulinoma cells trifluoperazine blocks Ca2+ influx-mediated insulin release and protein phosphorylation with no effect on basal or cAMP-mediated insulin release and protein phosphorylation with no effect on basal or cAMP-mediated secretion. Inhibition of Ca2+ influx-mediated insulin release and protein phosphorylation occurs with nearly identical dose dependence. Inasmuch as trifluoperazine affects voltage-dependent Ca2+ uptake in insulinoma cells, an involvement of calmodulin cannot be directly inferred. The evidence suggests that protein phosphorylation may be involved in the activation of the secretory apparatus by both cAMP and Ca2+. It is proposed that stimulation of insulin release by cAMP and Ca2+ is mediated by cAMP-dependent protein kinase and calmodulin-dependent protein kinase, respectively. PMID- 6281082 TI - Calmodulin and Ca2+ in normal and transformed cells. AB - Numerous lines of evidence implicate calcium and calmodulin (CaM) as regulators of cell growth and functional differentiation. In light of this evidence, several studies of the possible involvement of the CaM system in cellular transformation by RNA and DNA tumor viruses have been carried out. This paper summarizes the evidence linking calcium and CaM to the regulation of cell growth and critically examines the evidence that increases in CaM levels occur in transformed versus normal cells. A nontraumatic method for synchronizing both normal and transformed chick fibroblasts is presented. This method is utilized in a comparison of CaM level throughout the cell cycle of Rous sarcoma virus transformed and normal chick embryo fibroblasts. These studies best support the hypothesis that the observed differences in CaM levels between transformed and normal cultures under optimal growth conditions may largely reflect differences in the proportion of cells in a dividing versus a nondividing state. PMID- 6281083 TI - Calcium: calmodulin and cancer. AB - When several fast-growing Morris hepatoma tissue lines are compared with normal adult liver tissue, the following observations are made: calmodulin activity is increased in the cytoplasm and decreased in the membrane of the tumor cells. Total calcium is increased three- to fivefold in the tumors. Cyclic AMP phosphodiesterase activity is increased, whereas cyclic GMP phosphodiesterase activity is decreased. In addition, several of the fast-growing Morris hepatoma tissue lines have a new calcium-binding protein that is not observed in adult liver tissue. It is probable that the Ca2+-calmodulin complex is very active in these rapidly growing tumors. PMID- 6281084 TI - Stimulus properties of opioids with mixed agonist and antagonist activity. AB - The discriminative stimulus properties of opioids with mixed agonist and antagonist activity are heterogeneous in contrast to the relative uniformity of the stimulus properties of classical morphinelike agonists. Patterns of stimulus generalization to mixed agonist-antagonists are critically dependent on factors such as the particular drug used for training, dose of the training drug, and species of the experimental subject. Opioids can be divided into three broad categories on the basis of their patterns of stimulus generalization in rats and squirrel monkeys trained to discriminate saline from morphine, cyclazocine, or phenyclidine (PCP), and these categories predict the ease with which their stimulus effects can be blocked by nalozone or naltrexone. 1) Opioids that are generalized completely to morphine; stimulus effects are easily antagonized. 2) Opioids that are generalized completely to cyclazocine; stimulus effects are antagonized with difficulty. 3) Opioids that are generalized completely to cyclazocine and PCP; stimulus effects not antagonized. The diversity of the stimulus properties of opioids is consistent with evidence that multiple populations of receptors subserve the actions of morphinelike agonists and agonist-antagonists. However, the stimulus effects of opioids in group 3 appear to be mediated by the same neuronal substrates that are acted on by PCP rather than by neuronal sites traditionally associated with the activity of opioids. PMID- 6281085 TI - Intrauterine adhesions: an updated appraisal. PMID- 6281086 TI - Differential response to adrenocorticotropin hormone stimulation in polycystic ovarian disease with high and low dehydroepiandrosterone sulfate levels. AB - The relative contributions of the ovary and the adrenal gland to androgen overproduction in polycystic ovarian disease (PCOD) remain controversial. In this investigation, patients with proven PCOD were divided into two groups, (1) those with low dehydroepiandrosterone sulfate (DHEAS) levels and (2) those with high DHEAS levels, and compared with controls for their response to adrenocorticotropin hormone (ACTH) stimulation. Significant differences in weight, degree of menstrual disturbance, and basal progesterone levels distinguished the two groups with PCOD. Although no discrete enzyme block was unmasked by ACTH, marked differences in steroid production ratios were apparent between the low and high DHEAS PCOD groups. These results suggest that in PCOD with high DHEAS (1) substantial differences in adrenal steroidogenesis pathways occur, (2) increased progesterone as well as inappropriate estrogen feedback may contribute to chronic anovulation, and (3) serum DHEAS levels may be a helpful screen in discerning those patients who have a significant adrenal component to their hyperandrogenism and may benefit from adrenal suppression alone or in combination. PMID- 6281087 TI - Leydig cell hypofunction resulting in male pseudohermaphroditism. AB - An 11-year-old patient with male pseudohermaphroditism presented with essentially normal-appearing female external genitalia. When examined, inguinal gonads, redundant foreskin, and some posterior labial fusion were found. Evaluation revealed basal testosterone (T) levels ranging from 65 to 107 ng/dl with slightly elevated serum gonadotropin levels (luteinizing hormone [LH]: 76 ng/ml, and follicle-stimulating hormone [FSH]: 568 ng/ml). Neither T nor its precursors increased with human chorionic gonadotropin (hCG) stimulation. However, progesterone (P), 17-hydroxyprogesterone (17-OHP), and cortisol (F) responses to adrenocorticotropic hormone (ACTH) were normal. Androgen binding and 5 alpha reductase activity in cultured genital skin fibroblasts were normal. These data, plus the microscopic finding of a markedly reduced number of Leydig cells, strongly suggest that the male pseudohermaphroditism in this patient was due to inadequate Leydig cell function unrelated to LH receptors. PMID- 6281088 TI - [Inhibitory systems in brain structures]. PMID- 6281089 TI - [Effect of lithium chloride on cholinoreactive brain structures]. PMID- 6281090 TI - [Effect of cationic surface-active agents on neuromuscular transmission in smooth muscle]. PMID- 6281091 TI - [Postactivation changes in the ultrastructure of motor neurons]. AB - The prolonged high-frequency synaptic activation of the frog spinal motoneurons suppressing their firing, leads to an increase in the size of synapses and to a considerable shift in their ultrastructure. Restoration of the functional and morphological alterations in the cells after cessation of their rhythmical activation proceeds with different speed: the restoration of neurons' ability to generate synaptic and spike potentials in response to testing stimulation of afferents occurs much earlier than the restoration of the cells' ultrastructure and size. Neurons with obviously changed ultrastructure of soma and synaptic apparatus are capable of generating synaptic and spike potentials in response to stimulation. At the same time, the size of the neurons returns to normal either than the normalizing of cell ultrastructures whose speed of restoration varies as well. PMID- 6281092 TI - [Regulation of the cellular action of antidiuretic hormone]. AB - The steps of cell reactions which could modulate the effect of the antidiuretic hormone (ADH) were investigated in experiments on frog urinary bladder. Adrenaline and D2O reduced the interaction between ADH and its receptors. The urinary bladder cells released an inhibitor of ADH changing the reaction of receptors to ADH; adsorption of this inhibitor increased the water permeability after addition of ADH. Increased intracellular concentration of cellular near basolateral membranes produced the increase of water permeability whereas near the apical membranes calcium produced its decrease acting, perhaps, on microtubules. Swelling of the cells caused by ADH didn't change the reaction of these cells to ADH. Nevertheless, the cells swollen in hypotonic solution before the application ADH showed a lesser reaction to ADH. The role of cAMP phosphodiesterase, hyaluronidase, aldosterone, prostaglandins and other physiologically active substances in the action of ADH has been discussed. The data obtained suggest some possible ways and mechanisms of regulation of the cellular action of ADH. PMID- 6281094 TI - [Effect of serotonin on nervous system function and behavior in the honeybee]. AB - Serotonin in concentration of 0.0025% increased the excitability of chemoreceptors and the dancing rhythm rate in honey bees; higher concentrations diminished electrical activity in the second thoracic ganglion, lowered dancing rhythm rate and accelerated the alteration of food conditioning reflexes. Serotonin exerted no effect on the excitability thresholds of the peripheral nervous activity. A possibility of serotoninergic mediation of neural event in disruption of genetically determined tryptophan metabolic pathways (tryptophanuria) is discussed. PMID- 6281093 TI - [Mechanisms of the adaptive and trophic effects of the sympathetic nervous system on brain vessels and tissue]. AB - Data on immediate and delayed effects of cervical sympathectomy and stimulation of cervical postganglionic sympathetic fibers on cerebral circulation, hypothalamic neurosecretion and cerebral metabolism, are presented. Acute experiments in cats revealed adaptive function of cerebral vascular sympathetic innervation during rapid changes of arterial pressure. Data on the influence of basal vascular tone on post-desympathization circulatory effects were obtained. Long-lasting consequences of sympathetic stimulation (vasoconstriction and cerebral oedema) seem to depend on the activation of vasopressin secretion. Biochemical studies in rats revealed changes of (K+-Na+) ATPase activity, degree of oxidation and phosphorylation as well as dynamics of cAMP-cGMP and prostaglandin E, F2 alpha concentration in brain tissue in different period after cervical desympathization in conditions of normal cerebral blood supply and after circulatory cerebral hypoxia. The significance of sympathetic innervation for the adaptive and compensatory brain reactions to circulatory changes and damages is discussed. PMID- 6281095 TI - [Changes in transcapillary fluid exchange and the distensibility of small intestinal blood vessels upon adrenergic stimulation]. AB - In acute experiments on cats, the mean coefficient of capillary filtration (CFC) in denervated small intestine was 0.10 ml/min/mm Hg/100 g and the stretching ability of the intestinal venous vessels was 0.21 ml/mm Hg/100 g. Infusion of noradrenaline (5 micrograms/min) reduced the CFC by 33% and the stretching ability by 24%. Blockade of vascular alpha-adrenoreceptors abolished and beta adrenoreceptors augmented the noradrenaline effect on CFC and stretching ability. The data obtained indicate presence of both alpha- and beta-adrenoreceptors in precapillary sphincters and venous vessels of small intestine. Stimulation of the alpha-adrenoreceptors entails constriction of precapillary sphincters and decreases venous stretching ability whereas stimulation of beta-adrenoreceptors leads to opposite effects. Noradrenaline exerts main effect on alpha adrenoreceptors of precapillary sphincters and venous vessels in the denervate intestine; alpha-blockade is followed by an obvious beta-effect in the form of relaxation of precapillary sphincters and increase of the venous stretching ability. PMID- 6281096 TI - Synopsis of endogenous opiate research. PMID- 6281097 TI - Changes in levels of plasminogen activator activity in normal and germ-cell depleted testes during development. AB - Levels of plasminogen activator activity were determined in testes obtained from normal and irradiated rats in various ages. During normal development, plasminogen activator activity per g testis increased rapidly between 40 and 60 days of age, but a comparable rise did not occur in germ-cell depleted testes of irradiated rats. Levels of enzyme in various populations of testicular cells were highest in Sertoli (varying between 1800 and 6300 units/mg protein in cell maintained under different culture conditions), and lowest in peritubular myoid cells (about 1 unit/mg protein), with intermediate levels in germinal cells (ranging between 147 and 560 units/Mg protein in residual bodies, spermatocytes and spermatids). No protease inhibitor could be detected in germ-cell extracts. The addition to the medium in which Sertoli cells were in culture of particles which can be phagocytosed (autoclaved E. coli) resulted in an increased formation of plasminogen activator activity by Sertoli cells. A synergistic enhancement of enzyme production resulted following the addition of submaximal quantities of dibutyryl cyclic AMP and autoclaved bacteria to sertoli cells in culture. On the basis of these data, we suggest that the presence of advanced germinal cells during gonadal development may stimulate the synthesis of plasminogen activator by Sertoli cells, mediated in part by the phagocytosis of residual bodies by sertoli cells which occurs prior to spermiation. PMID- 6281099 TI - Calcium requirement for alpha-MSH action on tail-fin melanophores of xenopus tadpoles. AB - The role of Ca2+ in alpha-MSH action on melanophores was studied, in vitro, with a bioassay on ventral tail-fin pieces from tadpoles of Xenopus laevis. Melanosome dispersion induced by alpha-MSH required 1-2 mM extracellular Ca2+. Gradual lowering of the extracellular Ca2+ levels produced a concentration-dependent inhibition of the alpha-MSH response; complete inhibition was obtained in a Ca2+ free medium containing 10-4 M EGTA. In Mg2+-free medium, normal dispersion was observed. The Ca2+ antagonists verapamil (10-4 M), methoxy-verapamil (10-4 M) and La3+ (10-3 M) inhibited the dispersion induced by 3 X 10-9 M alpha-MSH, whereas ruthenium red (10-3 M) was without effect. The ionophore A23187 mimicked the effect of the hormone. Melanosome movement per se was evidently independent of Ca2+, because cAMP and dibutyryl-cAMP induced a full dispersion in the absence of Ca2+. These results show that extracellular Ca2+ is specifically required for alpha-MSH action on tail-fin melanophores in vitro and suggests a Ca2+ influx concomitant with the action of the hormone. Possible intra- and extra-cellular Ca2+ sites are discussed. PMID- 6281098 TI - Relationship between stimulated prolactin release from GH cells and cyclic AMP degradation and formation. AB - We have studied the relationship between the prolaction (PRL) release induced by thyroliberin (TRH) and theophylline and the formation and inactivation of adenosine 3', 5'-cyclic monophosphate (cyclic AMP) in cultured rat-pituitary cells (GH3 cells). TRH, which stimulated prolactin release, increased cyclic AMP formation and stimulated transiently both the low- and high-Km cyclic phosphodiesterases. The maximal effect on the phosphodiesterase was observed at 30 mM TRH. The stimulatory effect of TRH on the activity of the cyclic AMP phosphodiesterases was duplicated by incubation of the cells with cyclic AMP (2 10 mM). In washed particulate GH3 cell fractions, TRH increased the adenylyl cyclase activity up to 180%. Treatment of GH3 cells with theophylline stimulated the release of PRL and inhibited cyclic AMP degradation probably leading to the measured increase in cellular concentrations of the nucleotide. The effects of TRH and theophylline on cellular cyclic AMP concentrations and on PRL release were additive. There was a positive correlation between PRL release and cellular cyclic AMP concentration (r = 0.97). The elevations observed in cellular cyclic AMP concentration after TRH treatment are due to increased formation which in turn leads to phosphodiesterase activation. Therefore, cyclic AMP formation appears to be an intermediary step in the stimulus-secretion coupling caused by the tripeptide. PMID- 6281100 TI - Calcium sites in MSH stimulation of xenopus melanophores: studies with photoreactive alpha-MSH. AB - Photo-affinity labelling of MSH receptors on tail-fin melanophores of Xenopus tadpoles with p-azidophenylalanine 13-alpha-MSH (Pap13)-alpha-MSH) or p azidophenylacetyl-serine1-alpha-MSH ([Apac-Ser1]-alpha-MSH) resulted in a long lasting stimulation of the melanophores which cannot be reversed despite continuous washing. The generation of this irreversible response is inhibited when photo-affinity labelling is performed in a Ca2+-free medium or in the presence of Ca2+ antagonists. The irreversible stimulation produced in normal medium is completely reversed upon removal of Ca2+ , but is not affected by Ca2+ antagonists or melatonin. Re-addition of Ca2+ after temporary removal restores to irreversible stimulation, even in the presence of Ca2+ antagonists or melatonin. This proves that covalent alpha-MSH-receptor complexes remain fully functional despite temporary deprivation of ca2+. Racemized alpha-MSH, which binds 'tightly' to the receptor and produces a long-lasting effect, is removed from the receptor in Ca2+-free medium, but not by Ca2+ antagonists or melatonin. These results confirm earlier results showing that at least 2 Ca2+ sites are involved in alpha MSH action, one associated with MSH-receptor binding and the other with the subsequent generation of the effect. The dual role of Ca2+ is not the result of EGTA present; it is specific (Mg2+ has no effect) and is identical for the two different photoreactive alpha-MSH derivatives. PMID- 6281101 TI - Plasminogen activator production by parietal endoderm cells: stimulation by a protein from pituitary. PMID- 6281102 TI - Down-regulation of epidermal growth factor receptors in mouse embryos. PMID- 6281103 TI - Decreased synthesis of large fucosyl glycopeptides during differentiation of embryonal carcinoma cells induced by retinoic acid and dibutyryl cyclic AMP. PMID- 6281104 TI - The development and use of microcarrier and glass sphere culture techniques for the production of herpes simplex viruses. AB - Two high productivity monolayer culture methods, the microcarrier and glass sphere culture, were compared for their capacity to support the growth of MRC-5 and Vero cells, and Herpes simplex type 2 virus. Cell growth was similar in both systems giving yields of 14 X 10(5)/cm2 in microcarrier and 18 X 10(5)/cm2 in glass sphere cultures with yields of Vero cells being marginally better than MRC 5 cells. Virus yields were only slightly lower in these cultures than in small scale stationary cultures and confirmed the fact that MRC-5 cells produce twice as much HSV-2 as Vero cells, thus neutralising the growth advantage of these cells. Techniques for harvesting the virus in small volumes need to be improved in order to get a high titre suspension from the high capacity cultures. PMID- 6281105 TI - Herpes simplex vaccine using cell membrane associated antigen in an animal model. AB - Vesicles of plasma membrane, containing no detectable live virus, were prepared by treatment of Herpes simplex virus type 2 (HSV-2) infected Vero cells with dithiothreitol and formaldehyde. These preparations proved to be antigenic on injection into guinea-pigs and also protected these animals against intravaginal challenge with HSV-2. PMID- 6281107 TI - Production and potential use of monoclonal antibodies against polio viruses. AB - Lymphocyte hybridomas secreting monoclonal antibodies against different strains of polio virus type 1, 2, or 3 have been produced. For this purpose Balb/C mice were immunized with purified and inactivated virus suspensions and their splenocytes were fused with P3X63Ag8 mouse myeloma cells. Screening for antibody production was performed in an enzyme-linked immunosorbent assay (ELISA). Antibodies were produced either in cell culture or in Balb/C mice by passaging the hybridomas as solid or ascitic tumors, after they had been cloned at least three times by limiting dilutions in microtiter plates. Specificities of a number of these monoclonal antibodies were determined in the ELISA and in a neutralization test using different polio virus subtypes. The results indicate that for epidemiological studies monoclonal antibodies may prove to be very useful tools. Also the use of monoclonal antibodies for vaccine production (affinity chromatography; characterization of viral substructures) and routine vaccine control purpose (antigen quantification; neutralization of vaccine virus) seems attractive. Two of the neutralizing monoclonal antibodies against polio virus type 1, showed a selective immunoprecipitation with VP1, which suggests that VP1 is an important polypeptide for the induction of neutralizing antibody in vivo. PMID- 6281106 TI - Further developments in the quantification of small RNA viruses by U.V. photometry of sucrose density gradients. PMID- 6281108 TI - The use of highly concentrated purified (by a large scale method) and long term liquid nitrogen stored foot-and-mouth disease viruses for the preparation of vaccines: physico-chemical quality controls and potency tests after storage. AB - In 1974 a new industrial technique for concentration and purification of FMD virus was presented at the OIE Conference. The bulk inactivated virus from this technique was stored in liquid nitrogen vapour until required for vaccine formulation. In 1981, having applied this technique regularly for seven years, we now describe the results obtained and the advantages gained in the field of trivalent O, A, C bovine vaccine production. Vaccines prepared from such bulk virus stocks after several years storage give good protection against virulent virus challenge. PMID- 6281109 TI - Observations and implications of proteolysis in preparations of foot-and-mouth disease virus. AB - The integrity of the VP1 protein of foot-and-mouth disease virus was assessed by polyacrylamide gel electrophoresis (following storage at 4 degrees C of conventional tissue culture preparations and concentrated preparations of the virus. There was little evidence of VP1 degradation in tissue culture filtrates whereas considerable degradation was observed throughout a range of different concentrates. The use of the proteolytic enzyme inhibitor 'Trasylol' appeared to inhibit VP1 degradation in some virus preparations. Vaccination experiments with guinea pigs indicate that cleavage of O BFS 1860 virus by a range of proteolytic enzymes was always associated with a lowered stimulation of neutralising antibody and total antibody. Experiments with six other strains treated with trypsin gave similar results to those obtained with O BFS 1860. PMID- 6281110 TI - Structure and expression of the hepatitis B virus genome. AB - By fusion of the hepatitis B virus (HBV) surface antigen (HBsAg) gene to that of the E. coli lac Z gene carried by a phage lambda derivative, expression of HBsAg antigenic determinants was obtained and carried by a 138,000 dalton fusion polypeptide. Such a protein could be ultimately useful for second generation vaccine production. HBsAg gene expression was studied in eukaryotic cells using the mouse L cell (tk-) system. Cotransformation using a plasmid carrying two copies of the HBV genome in a tandem, head-to-tail arrangement (pCP10) and the cloned HSV-1 tk gene resulted in the excretion of 22 nm HBsAg particles in the supernatant. No other HBV markers were detected. These particles possess the same characteristics as the human serum particles (morphology, diameter, density, antigenicity). The purified HBsAg particles from L cells were found to be highly immunogenic in mice. HBV mRNA transcripts from these cells were analysed by Northern blotting. A major species of 2,300 bases was detected. This was mapped on the genome by hybridization with subgenomic fragments and in the L cell system using a series of plasmid derivatives carrying insertions at specific sites in the HBV genome and assaying for HBsAg expression. Thus the HBsAg gene promotor was localized between positions 2,400-2,800. Indeed there is only one TATA like sequence in this region, starting at position 2,776. PMID- 6281111 TI - Construction of a dominant selective marker useful for gene transfer studies in animal cells. AB - Biochemically transformed clones of murine, simian and human cells were obtained after transfection with a new dominant selective marker. This marker is a hybrid gene which was constructed with a bacterial neomycin-kanamycin resistance gene coding region and the transcription signals of the herpes simplex virus thymidine kinase gene. PMID- 6281112 TI - Detection and elimination of cellular nucleic acids in biologicals produced on continuous cell lines. AB - Experiments have been conducted to determine the extent to which currently available purification techniques can remove contaminating substrate cellular DNA from inactivated poliovirus vaccine produced on continuous cell lines rising highly [32P]-labeled, nick-translated cellular DNA added to poliovirus suspensions, we found that purification procedures were capable, in small-scale experiments, of reducing contaminating DNA by factors of 10(3) (DNAse treatment followed by gel filtration) and 10(3)-3X10(5) (ion exchange chromatography). Sequential application of these purification steps should reduce contaminating cellular DNA to acceptable levels. We also examined the potential usefulness of immobilized nucleic acid hybridization techniques for the routine direct testing of residual cellular nucleic acids in final production lots of inactivated poliovirus vaccine and other biologicals. A filter hybridization test, using [32P]-labeled, nick-translated cellular DNA as a probe, was capable of detecting 40 pg of homologous cellular DNA. Using probes of higher specific activity the assay should be sensitive enough for use in routine quality control. PMID- 6281113 TI - [An inhibitor of the replication of varicella-zoster virus]. PMID- 6281114 TI - [Clinical experience with the use of a new cephalosporin (cefotaxime) in infectious bronchopulmonary pathology]. PMID- 6281115 TI - Expression of herpes simplex virus common surface antigens and malignancy by clonal cells of a herpes simplex virus type 2-transformed line. AB - Herpes simplex virus (HSV) type common surface antigens (CSA) were examined by indirect immunofluorescence with rabbit antiserum to HSV type 1 in a clonal hamster cell line 155-4 transformed by HSV type 2. The tumor formation was examined in hamsters transplanted with various transformed and tumor cells. The examination of subclones derived from 1554-cell line gave the following results. (1) Thirty subclones were isolated and classified into three phenotypes as to CSA expression: (i) in CSA-positive type (20% of the clones isolated), the number of CSA-positive cells increased soon (5 hr) after seeding at 37 degree; (ii) in CSA inducible type (33% of the clones), the number of CSA-positive cells increased after treatment with actinomycin D (ACT-D, 2 micrograms/ml), but not without ACT D; (iii) CSA-negative type (47% of the clones), the number of CSA-positive cells did not increase after seeding or after ACT-D treatment. (2) In tumor cell lines derived from the parent line and from three representative clones, CSA expressions were similar to that by CSA-negative type. (3) Transformed cells expressing CSA after seeding or after ACT-D treatment formed tumors and metastases less efficiently than cells expressing little CSA in transplanted hamsters. (4) Tumor cell lines formed tumors and metastases more efficiently than transformed cells expressing little CSA in transplanted hamsters. PMID- 6281116 TI - Synthesis and antitumor effect of new biological alkylating agents, isethionic acid esters. AB - New hydrophilic alkylating agents, isethionic acid esters, are proposed for use as synthetic biological alkylating agents. Methyl, ethyl, and isopropyl esters of isethionic acid were synthesized starting from isethionate and the corresponding alkyl bromides or iodides in good yields. This synthetic procedure might be generally applicable to syntheses of alkyl isethionates. The derivatives thus prepared were water-soluble, as expected, and their alkylating abilities were very similar to those of the corresponding methanesulfonates. Hence, isethinonic acid esters might be suitable for use as hydrophilic biological alkylating agents in place of methanesulfonates. In order to determine the effectiveness of isethionates as anticancer alkylating agents, 1,4-butanediol diisethionate was prepared as a model compound and its anticancer activities against adenocarcinoma 755, sarcoma 180, L1210, and P388 were compared with those of the corresponding methanesulfonate, busulfan. The isethionate was superior to busulfan in all the assay systems employed. 1,5-Pentanediol diisethionate was also prepared and assayed. The results were similar to those for the 1,4-butanediol analog. In conclusion, in the design of molecules for use as cancer chemotherapeutics, the isethionic acid ester group is worth considering, and may be preferable to other commonly used leaving groups, including methanesulfonic acid ester. PMID- 6281117 TI - Lysozyme in Schwartz and Moloney virus-induced lymphoblastic leukaemia. AB - Studies have been made on lysozyme in serum and tissue homogenates in normal and leukaemic mice. An increase in serum lysozyme level occurred in mice bearing both strains of leukaemia as compared to the normal counterparts. Moreover, the elevated enzyme levels seem to be related to the rise in neutrophils in peripheral blood. In the spleen and kidney of leukaemic mice, the lysozyme level was found to be lower than in the normals, while the enzyme level remained more or less unaltered in the liver. The findings are discussed in relation to other reports in the literature. PMID- 6281118 TI - A significant role of proximity of HVJ-antigen in the generation of tumor immunity. AB - According to the blocking test for proximity of cell surface antigens, L1210 leukemia-associated antigen was not linked to molecules carrying H-2d antigen or murine leukemia virus-associated antigen, but there was a close association between HVJ-associated antigen and L1210 leukemia-associated antigen on the surface membrane of HVJ-pi-infected L1210 leukemia cells. Once L1210 leukemia cells were infected with HVJ-pi, their adsorptive power was found to be greater than that of uninfected cells by quantitative absorption analysis. These phenomena may lead to a better recognition by the host of L1210 leukemia associated antigen, followed by generation of tumor immunity. PMID- 6281119 TI - A T-cell line derived from normal human cord leukocytes by co-culturing with human leukemic T-cells. AB - A unique T-cell line, MT-2, was established from normal human cord leukocytes of a male infant by co-culturing with leukemic T-cells from a female patient with adult T-cell leukemia. MT-2 cells expressed receptors for sheep erythrocytes and complement and were reactive with anti-T-cell and anti-Ia sera. They were negative for Fc receptors, surface immunoglobulin, and Epstein-Barr virus nuclear antigen. Chromosomally, the MT-2 line was male and most cells were shown to have a normal diploid karyotype. The cultured cells were tumorigenic when transplanted into immunosuppressed newborn hamsters. PMID- 6281120 TI - Earlier appearance of murine mammary tumor virus-associated antigens in duct alveolus nodules induced by transplantation of fetal salivary mesenchyme into C3H mouse mammary glands. AB - Mammary epithelium of adult C3H mice in contact with fetal salivary mesenchyme took morphogenetically a salivary gland-like pattern of growth and developed a hyperplastic nodular mass at the site of transplantation. When murine mammary tumor virus-bearing C3H mice were treated in this way, early development and high incidence of mammary cancer were observed from these nodules compared to unstimulated mammary glands of the same host. Since tumor development, the viral antigens in these hyperplastic nodules were examined by means of the immunofluorescence technique. As early as 2 weeks after mesenchyme transplantation, the antigens were observed in the ductular or alveolar structure of these nodules, while surrounding normal tissues were not stained. Positive fluorescence was also observed in the mammary tumors that developed from these stimulated nodules. These results strongly suggested the involvement of murine mammary tumor virus in the early development of mammary cancer induced by fetal mesenchyme transplantation. PMID- 6281121 TI - Transformation of normal human cord lymphocytes by co-cultivation with a lethally irradiated human T-cell line carrying type C virus particles. PMID- 6281122 TI - Giant esophageal ulcer associated with cytomegalovirus. AB - A young, former homosexual, and narcotics-abusing male patient developed malaise, fever, lymphadenopathy, esophageal candidiasis, lymphopenia, and anergy progressing to severe odynophagia and a weight loss of 60 lb. Repeated endoscopic examinations revealed a progressively enlarging giant esophageal ulcer. Rigid endoscopic biopsy specimens and autopsy specimens demonstrated innumerable typical cytomegalovirus inclusions in the esophageal ulcer and no invasive candidiasis. Inclusions were also demonstrated in clinically silent smaller ulcers of the terminal ileum, colon, and adrenals. This case illustrates a further gastrointestinal manifestation of cytomegalovirus disease. PMID- 6281123 TI - What a difference an antigen makes. PMID- 6281124 TI - Cyclic guanosine monophosphate effects on nutrient and electrolyte transport in rabbit ileum. PMID- 6281125 TI - Correlation between pancreatic enzyme secretion and plasma concentration of human pancreatic polypeptide in health and in chronic pancreatitis. PMID- 6281126 TI - Human ileal ion transport in vitro: changes with electrical field stimulation and tetrodotoxin. PMID- 6281127 TI - Effect of progesterone pretreatment on guinea pig gallbladder motility in vitro. AB - Experiments were designed to examine the effect of chronic pretreatment with progesterone (2 mg/kg . day for 5 days) on the in vitro contractile response of gallbladder muscle strips from adult male guinea pigs. The muscle strips were challenged with either acetylcholine or the octapeptide of cholecystokinin and dose-response relationships were determined. The data were compared with dose response curves obtained from untreated control animals. progesterone pretreatment produced right-ward shifts in the acetylcholine and octapeptide of cholecystokinin curves that were characterized by significant decreases in the maximal contractile response. Serum progesterone concentrations in the pretreated animals were significantly increased over control levels. The data support the hypothesis that the sluggish behavior of the gallbladder during pregnancy and during the luteal phase of the menstrual cycle may be due, in part, to progesterone-related alterations in the contractile properties of gallbladder smooth muscle. PMID- 6281128 TI - Stress, behavior, and immunity: animal models and mediating mechanisms. AB - Research with animal model systems demonstrates a causal relation between stress and disease susceptibility. The physiological sequelae of stressful stimulation vary with type of stressor, chronicity, and perceived control. Different patterns of endocrine secretion are thus associated with specific behavioral parameters. The immune system is regulated at several levels: genetic, cellular, hormonal, and neuronal. Stress and behavioral factors can modulate both cell-mediated and humoral immunity by impacting on the latter three levels of regulation. Both corticosteroids and catecholamines, released as part of the response to aversive stimulation, profoundly inhibit immune responsivity by binding directly to the lymphocyte surface or by releasing secondary mediators that increase the function of suppressor lymphocytes. In addition, there are direct neuroanatomical connections between the hypothalamus and lymphoid organs. Subtle changes in experimental design have yielded significant differences in results that have increased understanding of intermediary mechanisms. Such research complements psychoneuroimmunological and behavioral epidemiological studies in humans. PMID- 6281129 TI - [Directed mutagenesis]. AB - Methods for generating mutations in preselected genes of prokaryotes and eukaryotes are reviewed. Directed mutagenesis is based on mutagenic treatment of genome fragments in vitro instead of a whole genome in vivo. The cloning methods make it possible to perform directed mutagenesis even using conventional nonspecific mutagens. Methods have been elaborated for selective modification of specific sequences in cloned genes, based on the specific DNA recognition properties of restriction endonucleases and complementary poly- and oligonucleotides. Chemically synthesized oligonucleotides may be used for constructing mutant genes containing any base substitutions, small insertions and deletions. PMID- 6281130 TI - Cloning and characterization of a genomic DNA fragment carrying the basic copy of the gene coding for variant surface antigen 118 of Trypanosoma brucei. AB - It has been proposed (Hoeijmakers et al., 1980b) that variant surface antigen (VSA) gene expression in Trypanosoma brucei is accomplished by a gene re arrangement involving the basic copy of the VSA gene to give the so-called expression-linked copy (which is present only in the strain expressing that particular antigen). In this publication, the basic and expression-linked copies of the gene have been visualized by Southern blot analysis of nuclear DNA and shown to be located on HindIII fragments of 4.5 and 10-12 kb, respectively. In addition, several other bands of weaker hybridization are seen, probably representing evolutionary relatives. Using a shotgun approach, HindIII gene banks have been constructed and recombinants isolated which carry the 4.5-kb HindIII fragment containing the VSA118 gene basic copy. Several clones containing evolutionary relatives were also found. The 4.5-kb HindIII fragment is able to hybridize to probes derived from both the 5' and 3' ends of the cDNA, while the relatives have homology only to the 3' end. A detailed comparison of the restriction map of VSA118 cDNA with that of the VSA118 basic copy showed no differences, demonstrating that the gene contains no introns. This result also indicates that the gene from which VSA118 mRNA is transcribed (whether this be the basic copy or the expression-linked copy) is identical to the basic copy over the region analysed. PMID- 6281131 TI - Efficient transformation of Serratia marcescens with pBR322 plasmid DNA. AB - Eight Serratia marcescens strains tested could be transformed with the plasmid pBR322. Transformants were selected on the basis of resistance to high levels of ampicillin (400 to 500 micrograms/ml). For six of the strains, the CaCl2- mediated transformation procedure developed for Escherichia coli was successful. For the other two strains, no transformants were obtained with the CaCl2-mediated transformation procedure unless the cells first received a heat treatment. Transformation frequency was dependent on DNA concentration, and no transformation was detected with linear pBR322 DNA. The stability and copy number of pBR322 were similar in S. marcescens and E. coli. As in E. coli, the pBR322 DNA was amplified in S. marcescens after inhibition of proteins synthesis. Based on these results, cloning in S. marcescens should be possible and pBR322 should be a useful cloning vehicle. PMID- 6281132 TI - A B1 repetitive sequence near the mouse beta-major globin gene. AB - A sequence which lies 2.8 kb to the 3' side of the BALB/c mouse beta-major globin gene has been identified by its ability to hybridise to a member of the human Alu repetitive sequence family. Nucleotide sequencing revealed a 133-bp region that shows 89% homology to the consensus sequence of the B1 family, the murine equivalent of the Alu family. To the 3' side of this sequence is a 31-bp region, C(A)3(C)2T(C)3G(C)11(A)9, which contains oligo(C) and oligo(A) tracts. The whole 164-bp sequence is flanked by a 16-bp imperfect direct repeat, G(A)4GGAGTCTCATAG. The orientation of the B1 sequence is such that transcription by RNA polymerase III would be expected to occur in the same direction as transcription of the neighbouring beta-major globin gene by RNA polymerase II. PMID- 6281133 TI - Human nucleotide sequences related to the transforming gene of a murine sarcoma virus: studies with cloned viral and cellular DNAs. AB - A recombinant plasmid, pI26, has been constructed by cloning into pBR322 a transforming gene of murine sarcoma virus (a Moloney strain, clone 124, MSV) synthesized by detergent-treated virions. From this plasmid a XbaI-HindIII fragment has been isolated which contains only mos-specific sequences. This mos specific probe has been used for screening a human gene library cloned in bacteriophage lambda Charon 4A. Of these, 19 clones have been isolated containing mos-related sequences. By physical mapping and molecular hybridization it has been shown that these sequences are neighboured by DNA regions related to Moloney murine leukemia virus. Recombinant phages have also been found containing human inserts related to MLV, not to the mos gene. The possible existence of murine like endogenous retroviruses in the normal human genome, including that of a sarcoma type, is discussed. By Northern blotting, expression of the cellular c mos gene has been detected in mouse liver treated with a hepatocarcinogen. The general significance of the suggested model for evaluating the relationship between chemical carcinogenesis and oncogene expression is discussed. PMID- 6281134 TI - Phasmids: hybrids between ColE1 plasmids and E. coli bacteriophage lambda. AB - Plasmids carrying cloned lambda att sites may be integrated into the bacteriophage genome by the site-specific recombination mechanism of lambda. The cross, referred to as "lifting" the plasmid, requires mixed infection of an Escherichia coli strain carrying the plasmid with two appropriately constructed "lifting" lambda phages. One phage donates a short left arm and the other donates a short right arm. These two short arms are of insufficient length to produce a viable phage genome and yield no recombinants when crossed on standard bacteria. However, viable recombinants are obtained when the genome length is extended by integration of one or more plasmids. We call these recombinants phasmids. They contain multiple att sites introduced at the ends of the integrated plasmids, and in the presence of integrase, recombination between these att sites can be exploited to effect release of the plasmid components. These novel genetic elements can be used in a variety of ways as vectors in genetic manipulation experiments. Sequences cloned in phasmids may be studied as a component of either a plasmid and or of a phage, and easily interconverted between the two states. PMID- 6281135 TI - High-level synthesis in Escherichia coli of the SV40 small-t antigen under control of the bacteriophage lambda pL promoter. AB - Several plasmids were constructed in which the SV40 small-t antigen gene was inserted in close proximity downstream from the thermoinducible leftward promoter (pL) of bacteriophage lambda. Upon temperature induction the best of our constructions expressed a small-t-related 19 000-dalton polypeptide in an amount corresponding to approx. 2.5% of total de novo protein synthesis. This 19 000 dalton protein was identified as small-t by specific immunoprecipitation with anti-T serum and by two-dimensional fingerprint analysis. In addition to the 19 000-dalton product, representative plasmids expressed fairly large amounts (up to 7% of total de novo protein synthesis) of a protein with an apparent Mr of 14 500. This 14 500-dalton polypeptide was shown to be related to authentic small-t. Presumably the secondary structure of the mRNA starting at pL is such that translation initiation at an internal AUG codon of the small-t gene is favored over initiation at the true initiating codon. PMID- 6281136 TI - Systematic alteration of the nucleotide sequence preceding the translation initiation codon and the effects on bacterial expression of the cloned SV40 small t antigen gene. AB - In the preceding paper (Derom et al., 1981) we described the cloning in bacterial plasmids of the simian virus 40 (SV40) small-t antigen gene under transcriptional control of the bacteriophage lambda pL promoter. Systematic variation of the distance and/or nucleotide sequence between the Shine-Dalgarno ribosome interaction sequence and the small-t translation initiation codon leads to considerable differences in production of small-t by the different plasmids. Secondary structure models derived for the different mRNAs confirm our previous conclusions about the requirement first for an accessible start codon and second for an accessible ribosome interaction site for efficient translation initiation. Secondary structure models for mRNAs from plasmids containing the small-t gene under control of the lac promoter are in agreement with these conclusions. PMID- 6281137 TI - Site-specific recA-independent recombination (fusion) of pMB8-related replicons in Escherichia coli in the region of the replication origins. AB - Escherichia coli recA+ and recA- cells were co-transformed with a mixture of pMB9 (Tcr) and pST8-26 (Apr, pBR325 derivative) plasmid DNAs followed by selection on plates containing both tetracycline and ampicillin. A set of stable Tcr Apr derivatives was isolated from these transformants. Many of the stable Tcr Apr segregants contained fused pMB9::pST8-26 plasmids with lengths that were about 0.8 kb longer than the sum of the lengths of the parental plasmids; one plasmid (pTF8) was about 1.5 kb shorter. The fusion was not stimulated by UV irradiation of co-transformants and occurred both in recA+ and recA- genetic backgrounds. Restriction analysis of the fused plasmids showed the two replicons were in the same relative orientation, and also indicated unique points of fusion in most cases (in 9 out of 10) which are localised within the 1.6-kb regions around the replication origins (RO). Because the fusion of plasmids of the type used in this study was not described before we have tentatively named it RO-fusion (Replication-Origin-fusion). PMID- 6281138 TI - Isolation and characterization of rat ribosomal DNA clones. AB - Four EcoRI fragments, which contain the transcribed portion of the rat rDNA repeat, have been isolated from a rat genome library cloned in lambda Charon 4A vector. Three of the fragments, 9.6, 6.7, and 4.5 kb, from clones lambda ChR-B4, lambda Nr-42, and lambda ChR-C4B9, contained part of the 5'-NTS, the 5'-ETS, 18S rDNA, ITS-1, 5.8S rDNA, 28S rDNA and approximately 3.5 kb of the 3'-NTS. Two EcoRI fragments, from clones lambda ChR-B4 and lambda ChR-B7E12, which coded for the 5'-NTS, the ETS, and most of the 18S rDNA, differed by 1 kb near the EcoRI site upstream of the 5' terminus of 18S rRNA. Restriction maps of the cloned DNA fragments were constructed by cleavage of the fragments with various restriction endonucleases and Southern hybridization with 18S, 5.8S, and 28S rRNA. These maps were confirmed and extended by subcloning several regions of the repeat in pBR322. PMID- 6281139 TI - Construction and characterization of new cloning vehicles. VI. Plasmid pBR329, a new derivative of pBR328 lacking the 482-base-pair inverted duplication. AB - The 4150-bp plasmid pBR329 was constructed by the the insertion into pBR327 of an 877-bp DNA fragment carrying the Cmr gene from pBR328. This new cloning vector does not contain the 482-bp inverted duplication that has been reported to be present in pBR325 and pBR328 (Prentki et al., 1981). In pBR329 the Cmr gene lacks its original promoter but is transcribed counterclockwise toward the Apr gene by a promoter located to the right of the HindIII site in the Tcr gene. PMID- 6281140 TI - Cloning of small DNA fragments containing the Escherichia coli tryptophan operon promoter and operator. AB - A41-bp AluI restriction fragment from the trp promoter-operator region has been cloned into the PvuII site of pBR322, regenerating PvuII sites. Transformants were selected on media that allowed the selection of trp-operator-bearing plasmids. The cloned 41-bp fragment can be released from the vector by PvuII digestion, and it possesses a functional promoter and operator as demonstrated by in vivo tests. The 41-bp fragment contains several restriction sites: HincII, TaqI, RsaI, and a HpaI site that is located at the center of the operator sequence. Two new operator derivatives, symmetrical about the HpaI site, were prepared from the 41-bp fragment by joining two right-side, or two left-side PvuII-HpaI pieces together at the HpaI site. These derivatives showed in vivo operator activity. Plasmids containing up to five copies of the 41-bp trp promoter-operator fragment have been constructed. These plasmids should be useful in preparing large amounts of the 41-bp fragment. PMID- 6281141 TI - Construction and use of SPP1v, a viral cloning vector for Bacillus subtilis. AB - A unique BamHI restriction site has been inserted into a nonessential region of the genome of a deletion mutant of phage SPP1. Construction of this phage, designated SPP1 v, required the in vitro conversion of a BclI site to a BamHI site. SPP1 v has been used as a vector phage to clone BamHI, BglII and BclI generated restriction fragments of DNA. A direct selection for recombinants has been developed. Transfection with SPP1 v requires intact, genomic-length molecules, and cleavage with BamHI destroys the transfecting ability of this DNA. Recombinants in which the BamHI site has been destroyed by ligation to Bg/II or BclI-generated fragments of DNA become resistant to BamHI digestion after ligation and are active in transfection. Cloning of DNA containing BamHI sites has been accomplished by using the enzyme Bst1503I to methylate BamHI sites before insertion, and so to protect them during the BamHI digestion used to select against vector molecules. The in vitro construction of SPP1 v generated XmaIII sites directly adjacent to, and on both sides of the inserted BamHI site. This permits precise excision of cloned DNA even when cloning destroys the BamHI insertion site. Restriction-enzyme generated fragments of DNA in the size range of 0 to 4 Md have been cloned, including a full-length copy of plasmid pUB110, almost the complete sequence of plasmid pBR322, and a sequence of DNA that carried the lambda cos site. PMID- 6281142 TI - [Potential use of guanidine-resistant Enterovirus strains in assessing the effectiveness of sewage treatment]. PMID- 6281144 TI - Changes in intestinal alkaline phosphatase activity in cholera toxin-treated rats. AB - It is conceivable that brush border enzyme activities of the intestinal mucosa will change when bacterial toxins are exposed to the intestinal microvillous membranes. The effect of cholera toxin on the activity of intestinal alkaline phosphatase in rats was therefore determined in the intestinal mucosa by the histochemical method as well as in intestinal lymph by using lymph fistulated rats. Activity of intestinal alkaline phosphatase in the intestinal mucosa and lymphatics changed biphasically after the oral administration of cholera toxin to rats. For the first three hours after the administration of cholera toxin it was depressed; it then increased and at eight hours reached a maximum. These changes in the activity of intestinal alkaline phosphatase were prevented by the administration of chlorpromazine, a known inhibitor of adenylate cyclase activity. PMID- 6281145 TI - Estrogen and progesterone receptors in tumors of the human ovary. PMID- 6281143 TI - Mitogen stimulation of peripheral blood lymphocytes of duodenal ulcer patients during treatment with cimetidine or ranitidine. AB - During a double-blind randomised clinical trial of cimetidine and ranitidine in the management of duodenal ulcer, the response of patients' peripheral blood lymphocytes to optimal mitogenic stimulation in vitro has been measured. Treatment with cimetidine, but not ranitidine, was associated with a significant increase in the proportion of peripheral blood lymphocytes responding to this optimal mitogenic stimulation. We conclude that these effects of cimetidine may not be mediated at classical histamine H2-receptors. PMID- 6281147 TI - [Inhibition of acid secretion with substituted benzimidazole. A new principle in ulcer therapy?]. AB - Substituted benzimidazoles inhibit gastric acid secretion stimulated by histamine, pentagastrin and vagal innervation. In contrast to anticholinergic agents and H2-receptor antagonists these compounds also block secretion induced by exogenous cyclic AMP suggesting a more peripheral site of action. Recent data show that these substances inhibit a H+/K+-ATPase which is localized at the secretory surface of the parietal cells and is generally regarded as the proton pump within the gastric mucosa. Because of the unique distribution of H+/K+ ATPase the inhibitory action of substituted benzimidazoles may offer a new therapeutic approach in hypersecretory states. PMID- 6281146 TI - [Hormone activity of various Brenner tumors]. PMID- 6281148 TI - [Hemorrhagic herpes encephalitis. A difficult differential diagnosis with computerized tomography]. AB - Herpes simplex encephalitis (HSE) is the most common sporadically appearing encephalitis in Central Europe. Differential diagnosis to brain tumors or spontaneous intercerebral hemorrhage is difficult. There are CT scan findings which are characteristic of HSE but there are no pathognomonic patterns. These characteristic findings are helpful in differential diagnosis to neoplastic or vascular processes. Thus, other diagnostic procedures (i.e. brain biopsy) to confirm diagnosis of HSE and effective therapy may be carried out in time. The difficulties in differential diagnosis are shown by the presented case. PMID- 6281149 TI - [Pathobiologic classification of depressed early gastric carcinoma (author's transl)]. PMID- 6281150 TI - [Studies on the vibration disease - an attempt of a new classification (author's transl)]. AB - In the first section of this study, a survey was made of 1,215 subjects who had been using mowing madchines. Of them 548 subjects without clinical symptomes were examined closely. Vibration threshold and maximal grasping power showed a normal distribution. Since the values decreased with an advancing age, limit values for each age range were calculated from respective probability vales. Advancing age did not affect skin temperature which showed a skewed distribution: limit values therefore were obtained from cumulative frequency distribution. In the second section, we investigated the actual conditions of the authorized patients with vibration diseases. Few severe cases were encountered, but adequacy of the conventional classification as to the degree of severity was questioned. Finally, an attempt was made to classify the disorder according to our new criteria of RNA system. The new system revealed that the vibration disease is classifiable into 7 types: R, N, A, RN, RA, NA and RNA. Single form such as R and N type appeared relatively earlier in life with shorter duration of exposure to vibrating machines. A few cases of A type were seen. Combination form such as RN, RA, NA and RNA type was the form of more severly deranged disorder, as judged from the age of onset, duration of exposure to vibrating machines, physical signs and also laboratory findings.Therefore, the new classification with RNA system appears more adequate in describing the pathosymptomatology of the vibration disease than the conventional method hitherto used. PMID- 6281151 TI - [The prevalence of antibody to hepatitis A virus in different age groups of Hokkaido inhabitants (author's transl)]. AB - Serum specimens drawn at random from Hokkaido inhabitants were tested for antibody to the hepatitis A virus (anti-HAV) by radioimmunoassay. The prevalence in different age groups was as follows, cord blood and newborn infants 44.4%, under one year 0%, 1-9 years 5.3%, 10-19 years 0%, 20-29 years 9.1%, 30-39 years 80%, 40-49 years 90%, and over 50 years 100%. This survey confirmed that antibody against HAV could pass through placenta and HAV had been very common virus by the 1950's in Hokkaido. PMID- 6281152 TI - Enhanced binding of retinoic acid to its cellular binding protein in Lewis lung tumor cytosol by alkaline phosphatase treatment. PMID- 6281153 TI - Specific binding sites on human blood platelets for plasma lipoproteins. AB - Scatchard analysis of the binding of homologous plasma low density lipoproteins (LDL) to human blood platelets shows the existence of a uniform class of saturable specific binding sites. Platelets from healthy donors bound 1470 +/- 640 molecules of LDL per platelet, the constant of association Ka = (6.2 +/- 2.2) X 10-7 l x mol-1. Binding kinetics, temperature dependence, and experiments with formaldehyde-fixed platelets showed that internalization of LDL (at least the labelled apoprotein moiety) did not occur to any considerable degree under the experimental conditions employed. Both very low density lipoproteins (VLDL) and high density lipoproteins 3 (HDL3) markedly inhibited the binding of LDL. In contrast to LDL, HDL3 bound to 3200 +/- 410 binding sites per platelet with a Ka = (9 +/- 1.7) X 10(7) l x mol-1. Additionally, the results of inhibition experiments using both LDL and HDL3 in combination gave evidence, that the sites for HDL3 binding were not identical with those for LDL binding and each inhibited binding of the other noncompetitively with reduction of the binding affinity and the number of available binding sites. VLDL bound to the platelet plasma membrane in a nonspecific-nonsaturable way. Possible significances of the presence of specific LDL receptors on the platelet plasma membrane for recognized functions of these blood elements are discussed. PMID- 6281154 TI - Production of a T-antigen-related protein in mammalian cells after stable transformation with a cloned SV40 gene fragment. AB - A recombinant plasmid based on pBR322 has been constructed which carries the replicator proximal early region of SV40 DNA, including the viral origin of replication (ORI). It lacks a major part of the tumour antigen 3'-coding region, the large T-antigen termination codon and the polyadenylation site. The recombinant plasmid was transferred together with the herpes simplex virus thymidine kinase gene, as a selectable marker into mouse LTK- cells. Integration and expression of the cloned SV40 gene fragment in TK+ transformants could be demonstrated by DNA restriction and blot hybridization and by immunofluorescence techniques. PMID- 6281155 TI - Monophasic synovial sarcoma, epithelioid sarcoma and chordoid sarcoma: ultrastructural evidence for a common histogenesis, despite light microscopic diversity. AB - Ultrastructural examination of six rare sarcomas--four monophasic spindle cell tumours, one epithelioid sarcoma, one chordoid sarcoma--has revealed marked similarities at the electron microscopic level despite widely divergent light microscopic appearances. These features consisted of: 1 the presence of two cell types, viz. a clear cell and a cell resembling the fibroblast; 2 pseudoglandular spaces with projecting microvilli or filopodia, and with related tight junctions; 3 an amorphous intercellular ground substance with focal condensation into recognizable basement membrane. The findings suggest a common maturation of these diverse tumours to synovial-like tissue, and support the proposal of Hajdu Shiu & Fortner (1977) that these be considered variants of synovial sarcoma. Published ultrastructural studies of synovial, epithelioid and chordoid sarcoma are reviewed in the light of these findings. The ultrastructural differentiation of synovial sarcoma from extraskeletal myxoid chondrosarcoma, chordoma and the spectrum of malignant spindle cell tumours is discussed. PMID- 6281156 TI - Placental site trophoblastic tumour (trophoblastic pseudotumour): a study of four cases requiring hysterectomy including one fatal case. AB - The clinico-pathological features of four patients with placental-site trophoblastic tumour (trophoblastic pseudotumour) are presented. One patient had the nephrotic syndrome associated with evidence of disseminated intravascular coagulation, with complete resolution after hysterectomy. In two patients the tumour extended beyond the uterus, and one of them died with many metastases in spite of intensive post-operative chemotherapy and 'second look' laparotomy. In three patients the tumour behaved as an actively infiltrative neoplasm resistant to chemotherapeutic regimes usually effective for choriocarcinoma. Serum HCG levels were relatively low compared with those of choriocarcinoma. Histologically the tumours were predominantly composed of mononuclear cells supported by a variable amount of vascular stroma and lacked the bilaminar structure characteristic of choriocarcinoma. Scattered cells stained positively with anti beta HCG and anti-alpha HCG antisera. Prior curettage was diagnostic in two of three cases. We did not find a clear correlation between mitotic activity and subsequent behaviour. Inflammatory cell infiltration and evidence of organisation around the tumour may be favourable prognostic indicators. We agree with a recent publication stressing the variable behaviour of this tumour, and emphasize the importance of serum HCG monitoring. Total surgical excision is usually feasible and in aggressive cases offers the best chance of eradication. We support the recent suggestion that 'trophoblastic pseudotumour' is an unsuitable name for a potentially lethal disease. PMID- 6281157 TI - Inherited tubule disorders. PMID- 6281158 TI - Cancer of the gallbladder: a review of forty-three cases. AB - This study presents the clinicopathologic findings 43 cases of cancer of the gallbladder. Particular attention was paid to the classification of various histologic types along with their distribution by age, sex, race, the presenting symptoms, the presence of gallstones, the occupational history, and the sites of metastases. The most common type was found to be differentiated adenocarcinoma, and the bulk of the cases occurred in elderly white females. Of note was the fact that in 79 per cent of the cases it was possible to identify underlying predisposing circumstances, either cholelithiasis or a positive occupational history. In light of the dismal prognosis associated with this malignant tumor, the identification of such predisposing factors provides an important avenue for the future investigation and prevention of this disease. In addition, in this series, a high rate of multiple primary malignant tumors of different tissues was note. This was attributable to the age and sex distribution of the cases, that is, their being primarily elderly and female. However, this subpopulation of multiple cancers was significantly different with regard to etiologic factors, and this may provide further clues for future preventive measures for this cancer. PMID- 6281159 TI - Histogenesis of salivary gland pleomorphic adenoma (mixed tumor) with an evaluation of the role of the myoepithelial cell. PMID- 6281160 TI - Chronic relapsing polyneuropathy associated with immunoblastic lymphadenopathy. AB - Two episodes of severe, relapsing sensorimotor polyneuropathy closely paralleled the course of systemic illness in a patient with immunoblastic lymphadenopathy. A review of the literature shows three other cases, although none with documented pathologic changes in muscles and nerves. Based on the pathologic findings in this patient, the peripheral neuropathy associated with immunoblastic lymphadenopathy can be classified under the group of relapsing inflammatory polyneuropathies associated with systemic disorders. Whether prednisone therapy helped this condition is uncertain. PMID- 6281161 TI - Vulvar intraepithelial neoplasia: the concept and its application. PMID- 6281162 TI - Fibrous histiocytoma of the orbit. A clinicopathologic study of 150 cases. AB - This is a clinicopathologic study of 150 cases of fibrous histiocytoma of the orbit. The tumors occurred in adults (median age, 43 years). The upper and nasal portions of the orbit were the most common sites of involvement. The most common signs and symptoms were proptosis (60 per cent), mass (46 per cent), and decreased vision (25 per cent). Based on the histopathologic features, the tumors were classified in three groups: benign (94 cases), locally aggressive (39 cases), and malignant (17 cases). The biological behavior correlated well with the duration of symptoms, the size and margins of the mass, and the histologic classification. The rate of recurrence was 31 per cent for the benign tumors, 57 per cent for the locally aggressive tumors, and 64 per cent for the malignant tumors. Follow-up data were obtained for 123 patients, with a mean duration of seven years. The ten-year survival of patients with benign, locally aggressive, and malignant fibrous histiocytoma was 100 per cent, 92 per cent, and 23 per cent, respectively. Nine patients died as a result of the tumor, six from local invasion of adjacent structures and three from metastatic disease. Fibrous histiocytoma is the most common primary mesenchymal orbital tumor of adults. The origin of the neoplasm is probably a primitive mesenchymal cell. Complete surgical excision appears to be the treatment of choice. PMID- 6281163 TI - Adenoid cystic carcinoma of the lacrimal gland: the clinical significance of a basaloid histologic pattern. AB - The authors reviewed 74 adenoid cystic carcinomas of the lacrimal gland, 54 of which had sufficient follow-up data for analysis of survival. Patients with a basaloid pattern in their tumor had a five-year survival rate of 21 per cent and a median survival of three years, whereas patients whose tumor contained no trace of a basaloid component had a five-year survival rate of 71 per cent and a median survival rate of eight years. Nonparametric statistical analysis revealed that this difference in survival was significant at the 0.0005 level. The authors propose that, in the future, pathologists label all adenoid cystic carcinomas as either "basaloid" or "nonbasaloid" and code each case accordingly. PMID- 6281165 TI - A comparative ultrastructural study of the normal lacrimal gland and its epithelial tumors. PMID- 6281164 TI - Histopathology of meningiomas and gliomas of the optic nerve. PMID- 6281166 TI - Fiber counting and analysis in the diagnosis of asbestos-related disease. AB - Analysis of numbers and types of asbestos fibers present in lung tissue may provide insights into the pathogenesis of asbestos-induced disease, as well as diagnostic information concerning the relationship of a given lesion to asbestos exposure. This type of analysis requires extraction of fibers and asbestos bodies from lung tissue, preferably by means of a digestion-and-concentration technique, and examination with a combination of electron optical techniques, including electron diffraction and energy-dispersive x-ray spectroscopy. The combination permits definitive identification of asbestos fibers. Asbestos bodies have been shown to contain asbestos no matter what population they are found in, but they appear to be of value in ascertaining unusual exposure only when present in very large numbers. Numbers of asbestos bodies markedly underestimate total numbers of fibers present in lung. In patients from the general population, the mean number of asbestos fibers is about 1 X 10(6)/g dry lung; of this number, more than 80 per cent are fibers of chrysotile less than 5 microns long. Patients in the general population who have pleural plaques have about the same total number of fibers, but their lungs contain about a 50-fold increase in long thin amphibole fibers of commercial origin. Patients who have asbestosis and most patients who have mesothelioma have 100 to 200 X 10(6) fibers/g dry lung; the grade of asbestosis appears to be related to total fiber content. Occasional patients may develop mesotheliomas with much smaller fiber burdens. Both benign and malignant pleural diseases appear to be closely related to the presence of long thin amphibole fibers. Analysis of pulmonary fiber burden suggest that asbestos related disease is not merely a matter of total numbers of fibers present, but that factors such as fiber type and size are equally important. PMID- 6281167 TI - Rhabdomyoblastic nature of cytoplasmic inclusions in malignant rhabdoid tumor. PMID- 6281168 TI - Immunocytochemical study of the glial fibrillary acidic protein in human neoplasms of the central nervous system. AB - The distribution of the glial fibrillary acidic protein (GFAP) was investigated in sections of 131 paraffin-embedded brain neoplasms obtained at surgery or at autopsy. The unlabeled antibody immunoperoxidase (peroxidase-antiperoxidase, PAP) method was used. Equally good results were obtained from 17-year-old material and from recent material derived at surgery or autopsy and fixed with Bouin fluid or phosphate-buffered formalin. The perikaryons and processes of reactive astrocytes showed the most intense stain for GFAP. Positive reaction to antibody against GFAP of varying intensity was demonstrated in astrocytomas of various grades of malignancy (32 of 32), glioblastoma multiforme (10 of 10), subependymal giant cell astrocytoma (1 of 1), ependymoma (2 of 10), subependymoma (4 of 4), and astrocytes in mixed neoplasms (8 of 8). In two neoplasms diagnosed as malignant astrocytomas and in four neoplasms diagnosed as glioblastoma multiforme, GFAP stain was limited to a few neoplastic cells. Usually the stain was more intense over processes than in perikaryons, with the exception of gemistocytic astrocytomas and the giant cells in glioblastoma multiforme, which showed an equally intense stain over perikaryons and processes. The periphery of Rosenthal fibers was intensely positive for GFAP. In astrocytic neoplasms the number of GFAP-positive cells and the intensity of the stain were inversely proportional to the degree of malignancy. In the following neoplasms the reaction for GFAP was negative: oligodendroglioma (3), oligodendroblastoma (1), medulloblastoma (3), medulloepithelioma (1), neuroblastoma (1), pineocytoma (1), typical teratoma of the pineal (1), fibrosarcoma (1), pituitary adenoma (2), craniopharyngioma (1), chordoma (1), chemodectoma of globus jugulare (1), metastatic carcinoma (17), and lymphoma (8). In one of 18 meningiomas, endogenous peroxidase activity was seen in mast cells. All meningiomas studied were negative for GFAP. In one of six neurinomas a positive reaction for GFAP was detected over processes. The authors concluded that the immunostain for GFAP is useful in the diagnoses of astrocytic neoplasms and of mixed gliomas. PMID- 6281169 TI - Acute renal failure and renal tubular squamous metaplasia following treatment with streptozotocin. AB - Nephrotoxicity, in the form of transient proteinuria, azotemia, abnormalities of tubular function, and acute renal failure, is the major toxic condition following administration of streptozotocin. The renal morphologic and ultrastructural abnormalities associated with streptozotocin remain poorly defined. We describe a patient with metastatic islet cell tumor of the pancreas who was treated with 16 weekly courses of 1 g/m2 of streptozotocin without marked change in renal function. Following a six-week hiatus without change in renal function, a single course of 1 g/m2 of streptozotocin was administered and resulted in acute renal failure. Light microscopic examination of the kidneys showed irregularly dilated renal tubules lined by low cuboid epithelium. The cells were pleomorphic and showed some mitoses. Nuclei were irregular and variably hyperchromatic. Electron microscopic examination disclosed large aggregates of fine microfilaments in the proximal convoluted tubules and collecting ducts. Microfilament aggregates were both free in the cytoplasm and membrane bound. Microfilaments were proved to be tonofilaments by the demonstration of keratin within the epithelium, using the immunoperoxidase method. These data suggest that squamous metaplasia may be an important part of streptozotocin renal toxicity, and the suggestion is made that they may be an antecedent of neoplastic change. PMID- 6281170 TI - Assigning the polymorphic human insulin gene to the short arm of chromosome 11 by chromosome sorting. AB - We have determined the subchromosomal location of the human insulin gene by analyzing DNA isolated from sorted human metaphase chromosomes. Metaphase chromosome suspensions were sorted into fractions according to relative Hoechst fluorescence intensity by the fluorescence activated chromosome sorter. The chromosomal DNA in each fraction was characterized by restriction endonuclease analysis. Initial sorts indicated that the insulin gene-containing fragment resided in a fraction containing chromosomes 9, 10, 11, and 12. Studies of cell lines that contained chromosome translocations permitted the assignment of the insulin gene to a derivative chromosome that contains portions of the short arm of chromosome II. Simultaneous sorting of the normal homolog from this small derivative chromosome separated the two different sized insulin gene-containing restriction fragments in this individual. These data indicate that the two restriction fragments represent insulin gene polymorphism and not duplicate gene loci. PMID- 6281171 TI - Characterization of a phosphoprotein phosphatase from goat testis. PMID- 6281172 TI - Delta-9-tetrahydrocannabinol & lung surfactant of rat. PMID- 6281173 TI - Role of peroxidase in melanogenesis--search for a control mechanism. PMID- 6281174 TI - Correlation between blood group isoantigens and sex chromatin in tumours of the breast. PMID- 6281175 TI - Original antigenic sin to influenza in rats. PMID- 6281176 TI - Elevated adenosine cyclic 3',5'-monophosphate levels in human macrophages following their interaction with Salmonella typhimurium. PMID- 6281177 TI - Insulin binding to cultured B- and T-lymphocytes. AB - Insulin receptors have been demonstrated on activated but not resting T lymphocytes. We compared insulin binding to several T- and B-cell tissue culture lines transformed by either radiation or lymphotrophic Herpes virus. Two T-cell lines derived from mouse lymphomas, one T-cell derived from the cotton top marmoset and one mouse B-cell line bound, at most, 3% of a tracer dose of [125I]insulin. In contrast, positive B-cell line (1605 L) derived from a leukemic cotton top marmoset, bound 40% of tracer [125I]insulin. The binding was of high affinity (1.9 nM-1), displayed analogue specificity characteristic of the insulin receptor and receptor number per cell was 16,600. The data show that a transformed B-lymphocyte binds substantial levels of insulin. The magnitude of such expression might be related to the fact that this B-cell line also expresses Epstein-Barr virus. PMID- 6281179 TI - Effect of prostaglandins on elicitation of anti-viral-cytolytic activity. AB - We have studied the modulating effect of prostaglandins (PG) on the elicitation of secondary anti-vesicular stomatitis virus (VSV) cytotoxic T-lymphocytes (CTL) with viral antigens. The results indicate that PGE1 and PGE2 both inhibit the induction of anti-VSV CTLs by ultraviolet-inactivated VSV. Consistent with the result was the augmentation of anti-VSV CTL activity when VSV-primed spleen cells were cultured in the presence of indomethacin. These results suggest that PGs may modulate anti-viral CTL responses. PMID- 6281178 TI - Immunochemical identification of products of the mitochondrial cytochromoxidase gene expressed in E. coli cells. AB - Bacterial clones containing hybrid plasmids with mitochondrial DNA insertions were obtained. Among them, a clone synthesizing immunoreactive protein with molecular weight 45,000 was discovered by means of antibodies against mitochondrial cytochromoxidase. The synthesis of the protein depended on orientation of the mitochondrial DNA insertion in the hybrid plasmid. The size of the protein was in excess of the coding capacity of the insertion, which speaks of chimeric nature of the protein. Absorption of the protein by the immunoglobulin fraction containing antibodies against cytochromoxidase resulted in a loss of antibody binding to cytochromoxidase subunit III. PMID- 6281180 TI - Elevation of adenosine 3' 5' cyclic monophosphate level by Aeromonas hydrophila enterotoxin. PMID- 6281181 TI - Effect of dietary fibre on oral glucose tolerance test. AB - Role of dietary fibre in lowering blood sugar levels has been studied in mongrel dogs. Pectin, wheat bran, guargum and Isabghol were fed with standard diet. Blood sugar levels were almost the same on 7th day (P less than .05). However, after 15 days pectin reduced blood sugar levels significantly (P less than .05), while other dietary fibre could not bring significant effect on blood sugar levels (P less than .05). PMID- 6281182 TI - Normocalcemic pseudohypoparathyroidism (Type II). PMID- 6281183 TI - Immunization of cattle with a variant-specific surface antigen of Trypanosoma brucei: influence of different adjuvants. AB - Nine different adjuvants were examined for their ability to potentiate the humoral and cell-mediated immune responses of cattle to a soluble glycoprotein antigen prepared from Trypanosoma brucei. Serological responses as measured by the Farr assay were best augmented by the oil-based adjuvants and saponin. Cell mediated immunity, as assessed by specific lymphocyte transformation in vitro, was enhanced by all oil-based adjuvants at different intervals after immunization. Results from a challenge infection of immunized cattle with the homologous clone of T. brucei and from neutralization tests indicated that protection against infection was better correlated with specific antibodies than with cell-mediated responses. From these considerations, and the absence of tissue reactions at the site of inoculation, saponin was considered more practical than the oil-based or bacterial adjuvants for the elicitation in cattle of antibodies to purified soluble antigens. PMID- 6281185 TI - Myeloperoxidase-mediated oxidation of methionine and amino acid decarboxylation. AB - The myeloperoxidase (MPO)-mediated decarboxylation of amino acids and the MPO mediated oxidation of methionine, two potential bactericidal mechanisms, were compared. In the presence of the MPO system (MPO, 50 mU/ml; H(2)O(2), 0.1 mM; Cl( ), 75 mM), 50% of alanine (0.1 mM) was decarboxylated, whereas only 5% of methionine (0.1 mM) was decarboxylated. In contrast, under similar conditions, 80% of methionine was oxidized to methionine sulfoxide. Once methionine was oxidized to methionine sulfoxide, it was decarboxylated (75%) by the MPO system. Methionine at 0.1 mM completely inhibited the decarboxylation of alanine, whereas alanine at a concentration 200 times that of methionine had no effect on the MPO mediated oxidation of methionine. Sodium azide, an MPO inhibitor, inhibited the decarboxylation of alanine and the oxidation of methionine to the same extent. Tryptophan markedly inhibited the oxidation of methionine, whereas histidine stimulated it. Alanine, glycine, and taurine had no effect. In contrast, all of these amino acids and taurine markedly inhibited the MPO-mediated decarboxylation of alanine. NaN(3), tryptophan, and methionine, which inhibited the MPO-mediated oxidation of methionine, also inhibited the killing of Staphylococcus aureus or Klebsiella pneumoniae by the MPO system; whereas histidine, alanine, and glycine, which did not inhibit the oxidation of methionine, had less or no effect on the killing of these two bacteria by the MPO system. Results suggest that methionine is preferentially oxidized to methionine sulfoxide by the MPO system. Once methionine is oxidized to methionine sulfoxide, it is then readily decarboxylated by the MPO system. The agent responsible for the oxidation of methionine may play an important role in the MPO-mediated killing of bacteria. PMID- 6281184 TI - Neutrophil response and function during acute cytomegalovirus infection in guinea pigs. AB - The mobilization and functional characteristics of polymorphonuclear leukocytes (PMNs) at the site of an inflammatory stimulus were studied during acute cytomegalovirus infection in guinea pigs. Weanling Hartley strain guinea pigs were inoculated subcutaneously with approximately 10(6) 50% tissue culture infective doses of virulent salivary gland-passaged guinea pig cytomegalovirus. The virus was uniformly present in bone marrow, buffy coat, and casein-elicited peritoneal exudate cells 5 to 7 days after the inoculation. The mean numbers of circulating PMNs in the animals were 2,862/microliters in uninfected controls and 880/microliters in infected animals. The total peritoneal PMNs recovered 14 h after casein injection were 491 X 10(6) and 237 X 10(6) in control and infected animals, respectively. The number of 50% tissue culture infective doses of guinea pig cytomegalovirus per 10(6) purified peritoneal PMNs was 10(2.17). Neutrophil O2 consumption was similar in infected and control animals in response to either stimulation by a neutrophil activator, phorbol myristate acetate, or phagocytosis of Staphylococcus aureus. However, the maximal rate of H2O2 release and the percent killing of S. aureus by peritoneal exudate cells from infected animals were significantly reduced compared with uninfected controls during acute infection. Granulocytopenia, a decreased mobilization of PMNs to the site of the inflammatory stimulus, a diminished cellular release of H2O2 in response to a bacterial stimulus, and a modest reduction in bacterial killing were demonstrated during experimental acute cytomegalovirus infection. Such reductions in granulocyte number and function at inflammatory sites during this type of infection could alter antimicrobial defenses. PMID- 6281186 TI - Application of RNase T1 one- and two-dimensional analyses to the rapid identification of foot-and-mouth disease viruses. AB - The analysis of several isolates of foot-and-mouth disease virus by RNase T1 fingerprinting of the 32P-labeled RNA is described. It has been shown that use of the 35S induced RNA instead of the virus particle RNA has two advantages. (i) About 40 times more radioactivity is incorporated into the induced RNA. (ii) The RNA can be prepared much more rapidly, thus increasing the value of the technique in rapid diagnosis. One-dimensional maps, in which the RNase T1 oligonucleotides are separated according to size, have been shown to provide a valuable screening method for distinguishing between viruses. Those viruses giving similar one dimensional maps also gave similar two-dimensional maps. The value of using the length of the polycytidylic acid tract of foot-and-mouth disease virus as a diagnostic tool is also discussed. PMID- 6281187 TI - Incubation of trypanosome-derived mitogenic and immunosuppressive products with peritoneal macrophages allows recovery of biological activities from soluble parasite fractions. AB - This report describes further attempts to define the nature of the parasite product(s) responsible for the extensive changes in lymphoid tissue in mice during infection with Trypanosoma brucei. As previously described, potent mitogenic and immunosuppressive effects are induced by a trypanosome-derived crude membrane fraction in vivo. There was no enrichment in these activities when purified parasite surface membranes were used. Mitogenic activity can be recovered from soluble trypanosome material only when it is incubated with peritoneal macrophages before transfer into syngeneic recipients. Thus, by encouraging association with a critical target cell, soluble parasite products can be studied, and their active components can be separated by conventional methods. Preliminary fractionation of high-spin trypanosome supernatant over Sepharose 4B confined the mitogenic activity to the high-molecular-weight fraction, which is a macromolecular complex of proteins, glycoproteins, and lipid. Extracted lipid from this material was able to significantly suppress a primary immunoglobulin G anti-sheep erythrocyte response. The activity was periodate sensitive and pronase resistant. The use of macrophages in vitro may be a general method whereby important biological activities lost as a result of fractionation procedures can be recovered and the active components studied in greater detail. PMID- 6281188 TI - Effect of murine cytomegalovirus infection on mitogen responses in genetically resistant and susceptible mice. AB - Suppression of the blastogenic response of spleen cells was found during murine cytomegalovirus infection of the genetically susceptible BALB/c and also the more resistant BALB.K strains of mice. These results were observed for both the T-cell mitogen concanavalin A and the B-cell mitogen lipopolysaccharide. As the viral inoculum was increased, there was greater immunosuppression within each strain, the time of maximum depression coinciding with peak virus titers in the spleen. Although both strains developed similar splenic virus titers and exhibited a similar decrease in the proportion of splenic T-lymphocytes, there was greater suppression of the mitogenic response during sublethal infection of the more susceptible BALB/c strain. The suppression could not be readily accounted for by the presence of suppressor cells or by a change in sensitivity to mitogen. The results suggest that the extent of immunosuppression induced by murine cytomegalovirus is determined in part by host genotype. PMID- 6281189 TI - In vitro amoebicidal activity of immune cells. AB - The amoebicidal activity of peripheral blood lymphocytes and spleen and peritoneal cells from hamsters vaccinated against or protected from hepatic amoebiasis and from those with hepatic amoebiasis was investigated. Peripheral blood lymphocytes and peritoneal and spleen cells from vaccinated or protected animals can kill trophozoites of Entamoeba histolytica in vitro. In contrast, spleen cells from infected hamsters showed no significant cytotoxic effect on the parasite. These data suggest that cellular immunity plays an important role in host defense against hepatic amoebiasis. PMID- 6281190 TI - Detection of hepatitis A antigen in human liver. AB - For the first time, hepatitis A viral antigen (HAAg) was shown in liver biopsy tissue from a patient in the acute phase of hepatitis type A by light and electron microscopy, using the peroxidase-antibody technique. Under light microscopy, the staining for HAAg appeared as a fine, granular reaction product, scattered throughout the cytoplasm of hepatocytes and sinusoidal lining cells. Standard thin-section electron microscopy revealed virus-like particles, 24 to 27 nm in diameter, in cytoplasmic vesicles of hepatocytes and Kupffer cells. By immunoperoxidase electron microscopy, HAAg was detected on particles aggregated within cytoplasmic vesicles of hepatocytes, thus demonstrating that the virus like particles (24 to 27 nm) are hepatitis A virus. The surrounding membrane of the vesicles was also positive for HAAg. The distribution patterns of HAAg in human liver were virtually identical to those described for experimentally infected marmosets. It is notable that most HAAg was detected within vesicles of liver cell cytoplasm, suggesting the possibility of vesicle-oriented morphogenesis of hepatitis A virus. PMID- 6281191 TI - Antigenic stimulation of T lymphocytes in chronic nononcogenic retrovirus infection: equine infectious anemia. AB - Equine infectious anemia is a chronic disease of horses caused by a nononcogenic retrovirus. Studies were undertaken to determine the types of cells involved in the in vitro lymphoproliferative response to viral antigens and the dynamics of this reaction. It was observed that reactive lymphocytes were present at unpredictable times in the peripheral blood of infected horses. This reaction was shown to be specific for the interaction of equine infectious anemia virus and T lymphocytes. Enriched B-lymphocyte populations did not divide when exposed to equine infectious anemia virus. Macrophages were depleted from the reaction by two methods: adherence to Sephadex and a combination of binding to Sephadex and adherence to complement-coated erythrocytes. Both methods reduced the number of monocytes, but only the combination of Sephadex and complement-coated cells removed the accessory cells needed for lymphocyte proliferation. We conclude that during the chronic stages of equine infectious anemia the number of antigen reactive T lymphocytes fluctuates within the peripheral blood and that these cells require a complement-binding cell for reaction. The relationship of these cells to the lymphoproliferative stages of this disease is discussed. PMID- 6281192 TI - Attachment and ingestion of bacteria by trophozoites of Entamoeba histolytica. AB - Entamoeba histolytica trophozoites were found to be very selective in their interactions with bacteria. Two principal mechanisms were shown to be responsible for these interactions. Certain bacteria, such as a number of Escherichia coli and Serratia marcescens strains which are known to contain mannose-binding components on their cell surface, bound to mannose receptors on the amoeba membrane. This attachment was markedly inhibited by alpha-methylmannoside (0.5%), especially when the incubations were done at low temperature (5 degrees C). Other bacterial species, such as Shigella flexneri and Staphylococcus aureus, which do not possess a mannose-binding capacity, attached to the amoebae, but only with the aid of concanavalin A or after opsonization of the bacteria with immune serum. In both types of attachment, between 40 and 100 bacteria bound per amoeba, and considerable ingestion of bacteria into amoeba vacuoles was observed at 37 degrees C. The attachment of opsonized bacteria to the amoebae does not appear to be mediated by Fc receptors since Fab' dimers obtained after pepsin digestion of immunoglobulin were capable of mediating adherence. Furthermore, preincubation of the amoebae with aggregated human immunoglobulin G or with heat-inactivated immune serum and EDTA did not inhibit the attachment of opsonized bacteria. The attachment of opsonized bacteria was markedly inhibited by N-acetylglucosamine containing glycoconjugates, such as peptidoglycan and chitin oligosaccharides, as well as by N-acetylgalactosamine. These results indicate that amoebae can attach and ingest bacteria either by using their membrane-associated carbohydrate binding protein or by having their mannose-containing cell surface components serve as receptors. PMID- 6281193 TI - Enzymatic modification of bacterial receptors on saliva-treated hydroxyapatite surfaces. AB - Certain properties of experimental pellicles formed by the adsorption of salivary components on hydroxyapatite surfaces change over time. To determine whether enzymes likely to be present in the oral environment could induce such changes, pellicles were treated with saliva which had been incubated for 18 h at 35 degrees C to promote the elaboration of microbial enzymes. This treatment markedly reduced the numbers of Streptococcus mutans MT3 and JBP and S. sanguis FC-1 and C5 cells which attached, but it had little or no effect on the attachment of S. mitis RE7, Actinomyces viscosus LY7 and CK-8, Bacteroides gingivalis 381, or B. melaninogenicus subsp. intermedius 581. Heating the incubated saliva at 60 degrees C for 30 min partially reduced its pellicle modifying activity, whereas heating at 80 degrees C for 30 min or 100 degrees C for 15 min completely eliminated such activity. This indicated that the saliva contained heat-labile substances, presumably enzymes, which could affect the pellicle receptors involved in the attachment of S. mutans and S. sanguis. Treatment of saliva-treated hydroxyapatite with commercially obtained enzyme preparations also affected bacterial attachment. Thus, treatment with galactose oxidase reduced the numbers of the S. mutans strains which attached, whereas treatment with neuraminidase reduced the adsorption of S. sanguis FC-1 but not that of S. sanguis C5. Treatment with beta-glucosidase preparations derived from almonds significantly reduced the attachment of all of the streptococcal strains studied, but, when subjected to isoelectric fractionation, the adherence inhibiting activity did not correlate directly with beta-glucosidase activity. Treatment of the pellicles with trypsin or eight other glycosidases did not affect streptococcal attachment. Exposure of the enzymatically modified pellicles to fresh saliva did not restore the streptococcal receptors. Collectively, the data suggest that some bacterial receptors in the pellicle coating of teeth can be modified by enzymes likely to be present in the oral environment, and these interactions may affect oral bacterial ecology. PMID- 6281194 TI - Murine cytomegalovirus-induced immunosuppression. AB - The mechanism of murine cytomegalovirus-induced immunosuppression was investigated by examining the roles played by lymphocytes and adherent cells derived from spleens of infected SWR/J mice. As few as 100 infected cells per spleen were correlated with complete abrogation of mitogen responses at 4 and 5 days after infection. In a series of cell mixing experiments it was shown that the deficiency in infected spleens was due partly to the adherent cells, which apparently secreted an immunosuppressive factor, and partly to the infected lymphocytes, which upon exposure to this factor could no longer respond to concanavalin A presented to them by normal adherent cells. PMID- 6281195 TI - In vitro effect of asbestos fibers on polymorphonuclear leukocyte function. AB - Incubation of chrysotile and amphibole asbestos fibers with normal human peripheral blood polymorphonuclear leukocytes (PMN) resulted in a significant stimulation of PMN metabolic activity and generation of toxic oxygen by-products as measured by chemiluminescence (CL). Although all asbestos fibers tested were cytotoxic to PMN, cytotoxicity and CL varied disproportionately with fiber type. Anthophyllite asbestos produced the greatest PMN cytotoxicity. It also depressed PMN phagocytosis of latex beads the most and induced the greatest PMN CL response of the fiber types examined. We postulate that asbestos-induced release of toxic oxygen by-products from PMN which have infiltrated into the pulmonary alveoli may contribute to disease pathogenesis in asbestosis. PMID- 6281196 TI - Modulation of polymorphonuclear leukocyte chemiluminescent response to the chemoattractant f-Met-Leu-Phe. AB - Upon the interaction between polymorphonuclear leukocytes (PMNL) and soluble or particulate matter, the cells become metabolically activated and produce chemiluminescence. To respond with chemiluminescence to the chemotactic peptide formyl-methionyl-leucyl-phenylalanine (f-Met-Leu-Phe), the PMNL required conditioning of the cells at 22 degrees C or higher temperature prior to stimulus addition. This was not required when phorbolmyristate acetate or opsonized yeast particles were used as stimulus. When cell samples were obtained from PMNL suspensions during 240 min from preparation, the chemiluminescence signal triggered by f-Met-Leu-Phe progressively increased. Scatchard plot analysis of the chemiluminescence data indicated that storage of the cells resulted in modulation of both the number of functional receptors and the affinity for the receptor. The importance of receptor function control in the inflammatory process is discussed. PMID- 6281197 TI - Differential cyst(e)ine requirements in human T and B lymphoblastoid cell lines. AB - In an effort to find exploitable metabolic differences between human T and B lymphoblasts, we have compared the ability of lymphocytes of varying phenotype to grow in cystine-deficient medium. Only 6 of 12 human lymphoblastoid cell lines tested were able to utilize homocysteine thiolactone or cystathionine in place of cystine for growth. This difference in growth requirements was unrelated to the rate of cell division, the presence of Epstein-Barr viral genetic material, or whether or not the cell lines derived from benign or malignant tissues. Rather, all B lymphoblastoid cell lines grew in homocysteine thiolactone- or cystathionine-containing medium, while the T and non-T, non-B lymphoblastoid cell lines did not. Normal human peripheral blood T and B lymphoblasts did not respond to mitogens in the homocysteine thiolactone or cystathionine medium, but developed the ability to utilize these cysteine precursors after stimulation with concanavalin A, protein A, or Epstein-Barr virus. The differences in cysteine requirements among T and B cell lines may reflect a fundamental difference in de novo protein-synthesizing capacity of the two cell types. PMID- 6281198 TI - Characterization of the carbohydrate moiety in a purified allergen preparation from the mite Dermatophagoides farinae and its importance for allergenic activity as tested by rast-inhibition method. AB - The monosaccharide composition of a purified mite allergen preparation was mainly found to be: mannose, galactose, glucose, N-acetylglucosamine and N acetylgalactosamine quantitated to 15% of dry material by means of methanolysis and GLC of trimethylsilylated methyl glycosides. Protein content was found to be 65%, determined by the Lowry method. Methylation analysis and periodate oxidation indicated the main polysaccharide material to be built of 1,6-linked hexosamine, with side chains having mainly galactose end groups. Attempts to obtain separate carbohydrate fractions by ion exchange chromatography of the extract on CM Sephadex and DEAE-Sephadex columns were not successful. The RAST-inhibition test showed a lower IgE binding capacity for periodate-treated mite preparation, where sugars containing vicinal diols have been oxidized, compared to an untreated mite preparation. PMID- 6281199 TI - Presence of fibronectin in cold precipitates of patients with cryoimmunoglobulinemia. AB - Fibronectin (FN) has been implicated in the cryoprecipitation of fibrin fibrinogen complexes. Preliminary observations have suggested that it may also play a role in cryoglobulinemia. In the present study 32 cryoprecipitates from patients with hepatitis, chronic active hepatitis, essential mixed cryoglobulinemia, and inflammatory bowel diseases were analyzed and screened for the presence of FN. Most cryoproteins of patients with essential mixed cryoglobulinemia and inflammatory bowel diseases were of the mixed IgM-IgG type and all contained FN. In contrast, most cryoproteins of patients with hepatitis were composed of polyclonal IgM and were devoid of FN. It is possible that FN characterizes and facilitates the formation of mixed cryoglobulins. However, its presence does not seem to be an absolute requirement for cryoprecipitation. PMID- 6281200 TI - Investigation of glucose-6-phosphatase in the liver of the human fetus. PMID- 6281201 TI - Frequent follow-up as data gathering and continued care with alcoholics. AB - THe clinical value of frequent follow-up interviews as a form of continuing care for alcoholics was investigated. Sixty-seven male subjects who had participated in an inpatient alcohol research treatment study were followed up monthly for 2 years and interviewed in depth at the end of 2 years. Almost all subjects felt that the frequent follow-up contacts had functioned for them as continued supportive care. The possibility that frequent follow-up interviews may serve to consolidate and increase gains made in treatment is discussed. PMID- 6281202 TI - Electrophysiologic and histologic studies in leprosy and some acrodystrophic neuropathies. AB - In vitro electrophysiologic and light microscopic studies were carried out on the sural nerve in six patients with non-leprous neuropathy with plantar ulceration and in six patients with various types of leprous neuropathy. In the non-leprous group (with congenital and acquired neuropathy) the abnormalities in the compound action potentials of the myelinated (large and small) fibers were usually more striking than those in the unmyelinated fibers potentials. In the leprous neuropathies, on the other hand, the three major fiber groups tended to be involved indiscriminately, the unmyelinated fiber potential being as liable to abnormality as the myelinated fiber potentials. Histologically the nerve fiber damage in the congenital neuropathies appeared to be moderate to gross involvement of the myelinated fibers without the prominent demyelination and degeneration/regeneration seen in leprosy and the other acquired neuropathies. Leprous neuropathy showed, besides fiber abnormalities, the simultaneous deleterious effects of inflammation and fibrosis. It is a matter for speculation whether disturbed conduction in the large myelinated touch-pressure mediating fibers contributes significantly to impaired pain perception (mediated by small fibers) and plantar ulceration in leprosy and other acrodystrophic neuropathies. PMID- 6281203 TI - Neuritis in pregnancy and lactation. AB - One hundred and forty-six women were studied during and after 153 pregnancies (31 healthy contacts: 34 pregnancies; 115 leprosy patients: 119 pregnancies). One healthy contact and 51 leprosy patients developed neuritis during the study period. All leprosy patients, including those who were considered to be cured and had stopped treatment, were at risk. Neuritis was accompanied by Type 1 and Type 2 lepra skin reactions and/or deterioration of the patients' leprosy status; this was particularly the case when neuritis was associated with nerve pain or tenderness (overt neuritis). Neuritis without nerve pain or tenderness (silent neuritis), preceded by the complaint of "rheumatism" and the clinical finding of enlarged peripheral nerves, was seen more frequently than overt neuritis (48:37 episodes). Insidious silent neuritis with loss of sensory and motor function during lactation was a particularly dangerous and hitherto undescribed risk of pregnancy. PMID- 6281204 TI - beta-Endorphin: isolation, amino acid sequence and synthesis of the hormone from horse pituitary glands. AB - Beta-endorphin has been isolated from equine pituitaries. Its amino acid sequence is identical to that of ovine, bovine and camel beta-endorphins except for substitution of the threonine residue at position 6 by serine. The equine beta endorphin has also been synthesized by the solid-phase method. In comparison with the human hormone, equine beta-endorphin was shown to possess 3 times the receptor-binding activity in rat membrane preparations and 1.6 times the analgesic potency in the mouse tail-flick assay. PMID- 6281205 TI - beta-Endorphin. Circular dichroism of synthetic human analogs with various chain lengths in methanol solutions. AB - The alpha-helix content of human beta-endorphin (beta h-EP) has been determined by circular dichroism (CD) in solutions ranging from 0 to 95% methanol in water. In addition, the CD spectra of beta h-EP-(1-30),-(1-29),-(1-28),-(1-27),-(1-26), (1-21) and -(1-15) have been examined in 90% methanol, and their alpha-helix contents estimated using parameters determined for this solvent. Addition of methanol to beta h-EP solutions brings about a noncooperative formation of alpha helix. An attempt to correlate secondary structure in methanol with biological and immunological activities showed limited direct correspondence, but may indicate the involvement of the tertiary interactions. PMID- 6281206 TI - Isolation and properties of beta-endorphin-(1-27), N alpha-acetyl-beta-endorphin, corticotropin, gamma-lipotropin and neurophysin from equine pituitary glands. PMID- 6281207 TI - Spin trapping of reactive uracilyl radicals produced by ionizing radiation in aqueous solutions. AB - The DNA base analogue 5-bromouracil (BU) reacts with hydrated electrons to produce a highly reactive uracil-5-yl (uracilyl) radical. The uracilyl radical was spin trapped by 5,5-dimethylpyrroline-1-oxide (DMPO) and t-nitrosobutane (tNB). DMPO solutions (0.01 M at pH = 7, N2 or N2O saturated) without BU were irradiated with 60Co gamma-rays and the resulting spin adducts produced by e-aq, H., and OH. were identified. The irradiation of alkaline (pH 12, N2 saturated) solutions of DMPO alone simplified the e.p.r. spectrum of spin adducts in that only the H. adduct was observed. Irradiated DMPO solutions (pH 12, N2 saturated) containing BU yielded a composite of two six-line e.p.r. spectra, attributed to uracilyl spin adducts that have two distinct steric configurations, and a background spectrum of H.-DMPO adducts which was removed by computer-subtraction. Uracilyl-tNB spin adducts were detected by first trapping uracilyl radicals in irradiated photobleached 10 M NaOH glasses at 77 K and then dissolving these glasses in pH-adjusted tNB solutions. PMID- 6281208 TI - Sindbis virus-induced ocular immunopathology. AB - The intraocular injection of the Sindbis virus in adult BALB/c mice produces a uveoretinitis with little or no central nervous system involvement. Ocular disease starts on the third day after infection and presents as a mild to moderate iridocyclitis and retinitis, usually accompanied by typical severe dysplastic changes of the retina. The inflammatory infiltrate consists almost exclusively of lymphocytes and histiocytes. Immunosuppression of the mouse with cyclophosphamide on the day after infection markedly reduces or eliminates completely the inflammatory response, suggesting that the virus itself is not cytopathogenic. In the normal host, the virus replicates within the eye for several days but is then completely eliminated by day 8 after infection. In the immunosuppressed animal, virus titers reach greater levels than in the normal animal and then fall, in step with the developing inflammatory response. It would appear that the immunologic mechanisms responsible for clearance of the viral infection from the eye also mediate the ocular disease. PMID- 6281209 TI - Overall distribution of glucagon-like immunoreactivity in the chicken retina: an immunohistochemical study with flat-mounts. AB - Pancreatic glucagon-like immunoreactivity (GLI) in the chicken retina was investigated by immunohistochemical methods with frozen sections and flat-mounts. Observations of the frozen sections showed that GLI is localized in the amacrine cells. Flat-mounts showed the GLI cells are evenly localized in the retina (n= or approximately 400/mm2) and composed to two types of cells--one is larger and the other is small. The former cell type occupies mainly the peripheral retinal region and the latter the central region. In addition, flat-mounts showed two types of GLI fiber plexuses in the inner plexiform layer; one is located in the area between laminae 2 and 3 and runs circularly and the other is located in lamina 1 and runs randomly. The relationship between GLI and cyclic AMP was also examined. an increase of the formation of cyclic AMP was caused by pancreatic glucagon in the chicken retina, suggesting that this peptide might have neurotransmitter or neuromodulator roles in the chicken retina. PMID- 6281210 TI - Kinetics of ocular herpes simplex virus shedding induced by epinephrine iontophoresis. PMID- 6281211 TI - A retrospective study of 100 cases of carcinoma of the lung. PMID- 6281212 TI - "What's new in medicine?" -- the patient. PMID- 6281213 TI - Familial neuropathy with liability to pressure palsies. Report of a case. AB - A case of hereditary neuropathy with liability to pressure palsies is described. The main histological findings in sural nerve were focal thickenings of myelin "sausages", "tomaculae"-and wide variability of internodal length. Numerous fibers with signs of remyelination were present, while there was little evidence of active demyelination. There were moderate axonal changes. Electrophysiological study revealed slowed conduction velocity in peripheral nerves. The possibility of a congenital myelin defect in this disease is discussed. PMID- 6281214 TI - Trithiozine polyneuropathy: clinical, neurophysiological and histopathological study of three cases. AB - 3 peptic ulcer patients, treated for a few weeks with Trithiozine, developed polyneuropathy. The histopathological patterns in the 2 patients in whom sural nerve biopsy was done presented wallerian degeneration of the axons of the myelinated fibers, especially those of larger caliber. The evidence for a iatrogenic toxic etiology is discussed. PMID- 6281215 TI - [Hereditary complement deficiencies]. AB - Complement deficiencies of all nine C-components have been observed. Hereditary defects of early components of the classical pathway - C1, C4, C2 - are often associated with diseases of the immuncomplex-type especially with systemic lupus erythematosus, dermatomysitis, vasculitis and nephritis. Deficiencies of C3 and C3b inactivator are linked to severe and recurrent bacterial infections. Patients with hereditary defects of the so-called late components, C5-C9, show increased susceptibility to recurrent disseminated infections by neisseria gonorrhoeae and meningitidis. The most frequent of the defects of the complement system is the hereditary deficiency of C1-inactivator which is associated with hereditary angioneurotic edema. In this paper the C-defects and their genetics are described and possible pathomechanisms are discussed. PMID- 6281216 TI - [Genetic aspects of blue rubber-bleb nevus syndrome and multiple generalized glomangiomatosis]. AB - Morphology, clinical findings and histological examination allow to recognize the solitary glomus tumor from the multiple glomus tumors and the blue rubber-bleb nevus-syndrome; however, the latter are more difficult to identify separately. They are transmitted as autosomal dominant conditions with good penetrance. With the hypothesis of a dual mutation system and that of the putative induction of paramutations the authors try to account for the variable expression of these conditions. It may be suggested that the existence of a common histological pattern, the multiple systematic, the multiple generalized tumors and the blue rubber-bleb nevus syndrome being perhaps the different expression of the same basic pathologic process. PMID- 6281217 TI - Young people and drugs. PMID- 6281218 TI - The "alternative therapy" controversy: facts are needed. PMID- 6281219 TI - Viral infections common to human and nonhuman primates. PMID- 6281220 TI - Prevalence of bovine leukemia virus infection in Florida. AB - A serologic study was undertaken to determine the prevalence of bovine leukemia virus (BLV) infection in dairy and beef cattle in Florida. Using the agar gel immunodiffusion test with a glycoprotein antigen 47.8% of 7,768 dairy cattle and 6.7% of 4,911 beef cattle were found to have antibodies to BLV. The prevalence of BLV antibodies increased significantly (P less than 0.0001) with increasing age. After data were adjusted for age, prevalence of BLV antibodies was significantly associated with dairy breed (P less than 0.05) but not with species (Bos taurus and B indicus) or sex. PMID- 6281221 TI - Clinical pharmacology of polymyxins. PMID- 6281222 TI - Risk factors related to the prevalence of infectious bovine keratoconjunctivitis. AB - A cross-sectional epizootiologic study was conducted by mailed questionnaire to determine how the prevalence of infectious bovine keratoconjunctivitis (IBK) related to selected predisposing causes. The prevalence of IBK in Missouri during the summer of 1978 was 4.97 cases/100 cattle, with 45% of respondents reporting cases of IBK. A higher prevalence of IBK was associated with calves and yearlings, Hereford and Hereford-cross cattle, backgrounding cattle operations, and herds with a history of IBK. Vaccination of mature cattle against infectious bovine rhinotracheitis was associated with significantly higher prevalence of IBK in calves. A significantly lower prevalence of IBK was associated with dairy cattle operations, older cattle, and the winter months. There was no association between the prevalence of IBK and locale, nutrient supplementation, pasture management, use of vaccines other than infectious bovine rhinotracheitis, parasite prophylactic measures, or fly control measures. PMID- 6281223 TI - Sodium bicarbonate and bicarbonate precursors for treatment of metabolic acidosis. PMID- 6281224 TI - Purification and properties of two gentamicin-modifying enzymes, coded by a single plasmid pPK237 originating from Pseudomonas aeruginosa. AB - A broad host range multiresistance plasmid pPK237, originating from Pseudomonas aeruginosa mediates high-level resistance to gentamicin and tobramycin. It was found to code for two gentamicin modifying enzymes, which from their substrate profile by radioenzymatic assay were characterized as aminoglycoside acetyltransferase AAC(3)-I and aminoglycoside adenylyltransferase AAD(2"). The two enzymes were studied after purification from an Escherichia coli K12 host. The two gentamicin-modifying enzymes coded by PPK237 were completely separated by DEAE chromatography. The purification (126 fold) of the acetyltransferase was achieved by (NH4)2SO4 precipitation, DEAE chromatography and affinity chromatography. The purification of the adenylyltransferase was performed by affinity chromatography directly after (NH4)2SO4 precipitation. Both purified enzyme preparations showed a single protein band on disc electrophoresis. The Km for gentamicin C1 of the acetyltransferase was 0.066 mM. The amino acid analysis of the acetyltransferase coded by pPK237 showed a different aminoacid composition than that of the gentamicin acetyltransferase AAC(3)-I purified by Williams and Northrop17). The acetyltransferase after DEAE chromatography is stable for many months at -20 degrees C, while the adenylyltransferase after purification is highly unstable; it shows enzymatic activity only in the presence of Mg++. PMID- 6281225 TI - Plasmid DNA in the erythromycin producing microorganism, Streptomyces erythreus NRRL 2338. AB - Streptomyces erythreus NRRL 2338, the erythromycin producing microorganism, contains extrachromosomal (plasmid) DNA. Four different plasmids, pSE1, pSE2, pSE4 and pSE6 present in the wild-type strain have characteristic mobilities on agarose gel electrophoresis, molecular weight and restriction endonuclease digestion patterns. Treatment of the wild-type strain with ethidium bromide or acridine orange gave two variants, one that could not convert erythronolide B to 3 (alpha)-mycarosylerythronolide B and another than produced 2 approximately 3 times more erythromycin A than the parental strain. Although the plasmid DNA profile of these two variants is different from the wild-type strain, it is not possible to conclude that any of the structural genes for erythromycin biosynthesis are located on the plasmids of S. erythreus NRRL 2338. PMID- 6281226 TI - Synergistic effects of a macrolide and a cell wall-affecting antibiotic on Pseudomonas aeruginosa in vitro and in vivo. 1. Combined effects of a macrolide and a peptide antibiotic. AB - Synergistic effects of peptide and macrolide antibiotics against Pseudomonas aeruginosa were investigated in vitro and in vivo. Synergistic effects were evaluated by estimating the number of viable bacteria at varying intervals in the logarithmic growth phase. These bacteria were treated concurrently with polymyxin B (PL) at the final concentration of 1.56 U/ml and with 9,3"-di-O acetylmidecamycin (MOM) at varying concentrations. Synergistic effect was observed when PL was used with MOM at 3.13 and 12.5 microgram/ml respectively. When MOM at 50 microgram/ml was used with PL, the viable bacterial count was reduced to below 1/300 of the control to which PL alone had been added. Thus, the synergistic effect was remarkable. Similar results were obtained when colistin methanesulfonate (CL) was used instead of PL. Subsequently, attempts were made to determine if this action could also be found in in vivo experiments using mice. PL or CL was injected intramuscularly and midecamycin (MDM) or MOM was administered once or repeatedly by the oral route. Simultaneously, Pseudomonas aeruginosa strain IFO 3455 was inoculated intraperitoneally to mice. In the case of treatment once or repeatedly using both PL and MDM or MOM, the survival rate of infected mice increased significantly compared to single treatment by PL alone. Thus, the synergistic effects were demonstrated in four experiments. (The significance levels for the experiments were P = 0.070, 0.015, 0.042 and 0.024). Similar results were obtained when strain No. 5 was used to infect mice (P = 0.0096, 0.0027). When CL and MOM were given to mice once prior to infection with strain No. 5, synergistic effects were obtained as well (P = 0.010, 0.034). PMID- 6281227 TI - beta-Lactamase inhibition by 6 alpha-thiocyanatopenicillanic acids. PMID- 6281228 TI - A study of the biliary excretion of mecillinam in patients with biliary disease. PMID- 6281229 TI - Combination of mezlocillin and azlocillin with cephalosporin antibiotics: cefoxitin, cefoperazone, cefotaxime and moxalactam. PMID- 6281230 TI - The tolC locus of Escherichia coli affects the expression of three major outer membrane proteins. AB - tolC mutants, which are resistant to colicin E1 and also highly sensitive to detergents and dyes, were shown to lack the OmpF outer membrane protein. There was little effect on transcription as judged by the use of an ompF-lac operon fusion strain, and the tolC effect was probably due to a post-transcriptional effect. The NmpC protein and protein 2 were also tolC dependent. PMID- 6281231 TI - Arginine metabolism in lactic streptococci. AB - Streptococcus lactis metabolizes arginine via the arginine deiminase pathway producing ornithine, ammonia, carbon dioxide, and ATP. In the four strains of S. lactis examined, the specific activities of arginine deiminase and ornithine transcarbamylase were 5- to 10-fold higher in galactose-grown cells compared with glucose- or lactose-grown cells. The addition of arginine increased the specific activities of these two enzymes with all growth sugars. The specific activity of the third enzyme involved in arginine metabolism (carbamate kinase) was not altered by the composition of the growth medium. In continuous cultures arginine deiminase was not induced, and arginine was not metabolized, until glucose limitation occurred. In batch cultures the metabolism of glucose and arginine was sequential, whereas galactose and arginine were metabolized concurrently, and the energy derived from arginine metabolism was efficiently coupled to growth. No arginine deiminase activity was detected in the nine Streptococcus cremoris strains examined, thus accounting for their inability to metabolize arginine. All nine strains of S. cremoris had specific activities of carbamate kinase similar to those found in S. lactis, but only five S. cremoris strains had ornithine transcarbamylase activity. PMID- 6281232 TI - Use of in vitro gene fusions to study the uxuR regulatory gene in Escherichia coli K-12: direction of transcription and regulation of its expression. AB - The uxuAB operon is composed of two genes coding for enzymes involved in hexuronate degradation. This operon is negatively controlled by the uxuR and exuR regulatory gene products. Fusions that brought lac gene expression under the control of transcriptional and translational signals within the uxuR gene were used to study uxuR regulation. These fusions were formed on plasmid cloning vectors constructed by Casadaban et al. (J. Bacteriol. 143:971-980, 1980). The transcriptional direction of the uxuR gene was deduced from the restriction pattern and the phenotypic properties of the new plasmids. The gene is transcribed counterclockwise on the standard Escherichia coli map, as is the uxuAB operon. Introduction in trans of a compatible plasmid carrying a wild-type uxuR gene in the lac fusion plasmid containing strain resulted in a decrease of beta-galactosidase synthesis. It was concluded that expression of the uxuR gene itself is repressed by its own product. The two other types of regulation found in the uxuAB operon, i.e., induction by fructuronate and catabolite control, also apply to the uxuR gene, whereas repression by the exuR repressor does not seem to occur for the uxuR gene. PMID- 6281233 TI - The BtuB group col plasmids and homology between the colicins they encode. AB - Colicins A, E1, E2, E3, E4, E5, E6, and E7 exhibited reduced activity against BtuB mutants of Escherichia coli K-12 and also against wild-type cells in the presence of vitamin B12. Plasmids encoding representatives of these colicins were specifically immune to high levels of the homologous colicin. Col(+) cells grown in media containing mitomycin C accumulated large amounts of colicin polypeptide. ColE2(+), ColE3(+), ColE4(+), ColE5(+), and ColE6(+) cultures also synthesized large amounts of second, lower-molecular-weight protein under these conditions. Colicins E2 through E7, but not A or E1, reacted with antiserum raised against purified colicin E3. Colicins E2 and E7 induced synthesis of beta-galactosidase encoded by lacZ under the control of the colicin Ib gene promotor on a derivative of Col plasmid ColIb.P9. This promotor is usually active only when the cells are treated with agents which damage DNA or block replication. Plasmids encoding various mutant forms of colicin E3 (M. Mock and M. Schwartz, J. Bacteriol. 142:384-390, 1980) recombined with ColE2, ColE4, ColE5, or ColE6 plasmids at a frequency of 10(-4) per cell to produce a colicin active against ColE2(+), E4(+), E5(+), or E6(+) cells. ColE5 and ColE6 plasmids recombined with ColE3 plasmids bearing mutations affecting colicin E3 receptor recognition, envelope penetration, and catalytic activities. ColE2 and ColE4 plasmids recombined only with ColE3 plasmids bearing mutations affecting receptor recognition and envelope penetration. Recombinants between mutant ColE3 plasmids and ColA, ColE1, or ColE7 plasmids were not detected. We propose the designation BtuB group for the colicins described here, and we divide the group into two classes comprising colicins A and E1, which act on the cytoplasmic membrane, and the related colicins E2 through E7, which have known or putative nuclease activities. PMID- 6281234 TI - Origin and direction of mini-R1 plasmid DNA replication in cell extracts of Escherichia coli. AB - Replication of the mini-R1 plasmids pKN177 and pKN182 can be carried out efficiently in cell extracts of Escherichia coli and depends on both transcription and translation. Heteroduplex and restriction analyses indicate that both plasmids are derived from the R1 copy mutant pKN104 by Is1-mediated recombination events without involving structural alterations in the replication region. To ascertain whether the in vitro replication of these miniplasmids corresponds to R1 replication in vivo, the origin and direction of replication were analyzed by electron microscopy of replicative intermediates. It was found that replication starts at a unique origin located within the RepA region at R1 coordinate at 82.4 kilobases and proceeds unidirectionally toward the IS/b sequence. The specification of the origin and the direction of in vitro replication are therefore in full agreement with the pattern observed previously for the in vivo replication of the closely related plasmids R100 and R6-5. This agreement provides additional evidence that R1 DNA synthesis in vitro employs the same replication mechanism as it does in vivo. PMID- 6281235 TI - Control of enzyme synthesis in the arginine deiminase pathway of Streptococcus faecalis. AB - The formation of the arginine deiminase pathway enzymes in Streptococcus faecalis ATCC 11700 was investigated. The addition of arginine to growing cells resulted in the coinduction of arginine diminase (EC 3.5.3.6), ornithine carbamoyltransferase (EC 2.1.3.3), and carbamate kinase (EC 2.7.2.3). Growth on glucose-arginine or on glucose-fumarate-arginine produced a decrease in the specific activity of the arginine fermentation system. Aeration had a weak repressing effect on the arginine deiminase pathway enzymes in cells growing on arginine as the only added substrate. By contrast, depending on the growth phase, a marked repression of the pathway by oxygen was observed in cells growing on glucose-arginine. We hypothesize that, in S. faecalis, the ATP pool is an important signal in the regulation of the arginine deiminase pathway. Mutants unable to utilize arginine as an energy source, isolated from the wild type, exhibited four distinct phenotypes. In group I the three enzymes of the arginine deiminase pathway were present and probably affected in the arginine uptake system. Group II mutants had no detectable arginine deiminase, whereas group III mutants had low levels of ornithine carbamoyltransferase. Group IV mutants were defective for all three enzymes of the pathway. PMID- 6281236 TI - Protein H encoded by plasmid CloDF13 is involved in excretion of cloacin DF13. AB - Excretion of cloacin DF13 was studied in Escherichia coli cells harboring different CloDF13 insertion and deletion mutant plasmids. Insertions of a transposon at position 9.8 or 11.5% of the CloDF13 plasmid blocked the expression of gene H and strongly reduced the specific excretion of cloacin DF13 into the culture medium, but had no effect on the production of cloacin DF13. Insertions in or deletions of regions of the CloDF13 DNA upstream the cloacin operon did not affect the excretion or production of the bacteriocin. Introduction of a CloDF13 plasmid that encodes for the gene H product in cells harboring a CloDF13 plasmid with an insertion in gene H stimulated the excretion of cloacin DF13 significantly in mitomycin C-induced and in noninduced cultures. Cloacin DF13 in cloacinogenic cells that did not produce the gene H protein was found to be about 90% located in the cytoplasm. In cells that did produce the gene H product, about 30% of the cloacin DF13 molecules were found in the cytoplasm, about 18% were found in the periplasm, about 2% were in the membranes, and about 50% were located in the culture supernatant. Cyclic AMP stimulated the production but not the excretion of cloacin DF13 in cells cultivated in the presence of glucose. PMID- 6281237 TI - Construction from Mu d1 (lac Apr) lysogens of lambda bacteriophage bearing promoter-lac fusions: isolation of lambda ppheA-lac. AB - Bacteriophage Mu d1 (lac Aprr) was used to obtain strains of Escherichia coli K 12 in which the lac genes are expressed from the promoter of pheA, the structural gene for the enzyme chorismate mutase P-prephenate-dehydratase. A derivative of bacteriophage lambda which carries the pheA-lac fusion was prepared; the method used is generally applicable for the construction, from Mu dl lysogens, of specialized transducing lambda phage carrying the promoter-lac fusions. A restriction enzyme cleavage map of lambda ppheA-lac for the enzymes HindIII and PstI is presented. PMID- 6281238 TI - Mapping of two ugp genes coding for the pho regulon-dependent sn-glycerol-3 phosphate transport system of Escherichia coli. AB - Two genes, ugpA and ugpB, coding for a binding protein-dependent sn-glycerol-3 phosphate transport system, were mapped at 75.3 min on the Escherichia coli chromosome. A Tn10 insertion in ugpA resulted in loss of transport activity but still allowed the synthesis of the sn-glycerol-3-phosphate-binding protein. This Tn10 insertion was found to be linked by P1 transduction to pit, aroB, malA, asd, and livH with 2.5, 2.8, 25, 63.5, and 83% cotransduction frequency. An insertion of Mud (Ampr lac) in ugpB resulted in the loss of the binding protein. ugpB is closely linked to ugpA. It is either the structural gene for the binding protein or located proximal to it. The analysis of the crosses allowed the ordering of the markers in the clockwise direction as follows: aroB, malA, asd, ugpA, ugpB, livH, pit. PMID- 6281239 TI - Energy coupling of facilitated transport of inorganic ions in Rhodopseudomonas sphaeroides. AB - Within the scope of a study on the effects of changes in medium composition on the proton motive force in Rhodopseudomonas sphaeroides, the energy coupling of sodium, phosphate, and potassium (rubidium) transport was investigated. Sodium was transported via an electroneutral exchange system against protons. The system functioned optimally at pH 8 and was inactive below pH 7. The driving force for the phosphate transport varied with the external pH. At pH 8, Pi transport was dependent exclusively on delta psi (transmembrane electrical potential), whereas at pH 6 only the delta pH (transmembrane pH gradient) component of the proton motive force was a driving force. Potassium (rubidium) transport was facilitated by a transport system which catalyzed the electrogenic transfer of potassium (rubidium) ions. However, in several aspects the properties of this transport system were different from those of a simple electrogenic potassium ionophore such as valinomycin: (i) accumulated potassium leaked very slowly out of cells in the dark; and (ii) the transport system displayed a threshold in the delta psi, below which potassium (rubidium) transport did not occur. PMID- 6281240 TI - Expression of an alternative nitrogen fixation system in Azotobacter vinelandii. AB - Nitrogenase activities were determined from maximum acetylene reduction rates for mutant strains of Azotobacter vinelandii which are unable to fix N2 in the presence of molybdenum (Nif-) but undergo phenotypic reversal to Nif+ under conditions of Mo deficiency. The system responsible for N2 fixation under these conditions is thought to be an alternative N2 fixation system (Bishop et al., Proc. Natl. Acad. Sci. U.S.A. 77:7342-7346, 1980). Phenotypic reversal of Nif- strains to Nif+ strains was also observed in N-free medium without Mo but with either V or Re. Two protein patterns were found on two-dimensional gels of proteins from the extracts of wild-type cells cultured in N-free medium without Mo and with or without V or Re. The expression of each protein pattern in the wild-type strain of A. vinelandii seemed to depend upon the physiological state of the N2-fixing culture. Electron paramagnetic resonance experiments were conducted on whole cells of A. vinelandii grown under conditions of Mo deprivation in the absence of fixed N. No g = 3.65 signal (an electron paramagnetic resonance signal characteristic of the Mo-containing component of nitrogenase) was detectable in these cells, regardless of whether V or Re was present during growth of these cells, These results are discussed from the perspective that the well-known effect of V on N2 fixation by A. vinelandii may involve an alternative N2 fixation system. PMID- 6281241 TI - Occurrence and properties of composite transposon Tn2672: evolution of multiple drug resistance transposons. AB - We found Tn2671 (the 23-kb long IS1-flanked r-determinant of NR1-Basel) inserted into the ampicillin resistance gene bla of the Tn3-related transposon Tn902. The resulting 28-kilobase-long composite transposon Tn2672 (= Tn902 bla::Tn2671) is stable, and it translocates as a unit into various loci including IS1 of the resistance transfer factor of R100-1. These results are discussed with respect to the evolution of R plasmids providing multiple drug resistance. PMID- 6281242 TI - Construction and behavior of strains with mutations in two chemotaxis genes. AB - Double mutants have been constructed by transducing each of the six che genes from the main che gene cluster into the cheC mutant with reversed behavior. The behavioral properties of these double-mutant strains were examined. The results are interpreted in terms of a model based on the cheC gene product being the component of the flagellar basal body that generates tumbling or smooth swimming in response to changes in the level of the response regulator. The properties of the double mutants can then be explained in ways which provide further understanding of the bacterial sensing system. PMID- 6281244 TI - "Frizzy" mutants: a new class of aggregation-defective developmental mutants of Myxococcus xanthus. AB - During fruiting-body formation in Myxococcus xanthus, cells aggregate into raised mounds, where they sporulate. A new class of aggregation-defective developmental mutants was identified within a collection of nonfruiting mutants of M. xanthus. The mutants failed to aggregate into discrete mounds, but rather aggregated into "frizzy" filaments. Many cells within the filaments sporulated normally. Pairwise mixtures of representative frizzy mutants were unable to stimulate each other to aggregate normally. Two strains of M. xanthus were isolated which contained transposon Tn5 insertions mapping near one frizzy mutation. A search through 36 mutants exhibiting the frizzy phenotype showed that all were linked to the same Tn5 insertion sites. Three-factor cross-analysis of 22 of these mutants allowed the mapping of these mutations into many loci. The localization of Tn5 inserts adjacent to this region make possible further manipulation of these genes. PMID- 6281243 TI - Enzymes related to fructose utilization in Pseudomonas cepacia. AB - Growth of Pseudomonas cepacia on fructose, mannitol, or sorbitol depended on formation of an inducible fructokinase (forming fructose-6-phosphate) and the presence of enzymes of the Entner-Doudoroff pathway. Mutants deficient in any of these enzymes failed to utilize the aforementioned carbohydrates. Fructokinase deficiency did not affect growth of the bacteria on glucose. Fructose was accumulated intracellularly by active transport. Mutants blocked in transport of fructose grew normally on mannitol or sorbitol despite their inability to utilize fructose. Growth on either of these hexitols or on galactitol was accompanied by induction of two hexitol dehydrogenases, one active primarily with mannitol and the other active with sorbitol and galactitol. As expected, a mutant deficient in mannitol dehydrogenase failed to utilize mannitol as a carbon and energy source but grew normally on sorbitol and galactitol. Extracts of bacteria grown on fructose, mannitol, or sorbitol and higher levels of phosphoglucose isomerase than extracts of bacteria grown on alternate carbon sources such as citrate or phthalate. The higher levels were due to appearance of a second phosphoglucose isomerase species not present in cells with the lower activity. The results indicate that the initial steps in fructose utilization by P. cepacia differ from those of most other pseudomonads, which transport fructose by phosphoenolpyruvate dependent translocation, forming fructose-1-phosphate, and suggest that degradation of fructose, mannitol, and sorbitol occurs primarily via the Entner Doudoroff pathway. PMID- 6281245 TI - Circulation of potassium across the plasma membrane of Blastocladiella emersonii: K+ channel. AB - A previous paper reported that the water mold Blastocladiella emersonii generates a transcellular electrical current, such that positive charges enter the rhizoid and leave from the thallus (Stump et al., Proc. Natl. Acad. Sci. U.S.A. 77: 6673 6677, 1980). To begin to understand the genesis of this current we investigated ionic relationships in this organism by use of intracellular microelectrodes. In cells suspended in buffered CaCl2, the membrane potential could be accounted for as a K+ diffusion potential; no evidence for an electrogenic pump was obtained. Potassium ions diffuse outward by a pathway that also carries Rb+ and Ba2+, but excludes both smaller and larger ions (Li+, Na+, Cs+, Mg2+, Ca2+, and choline). Chloride and other anions make little contribution to the potential, but the presence of Ca2+ in the external medium is required for successful potential measurements. In growing cells, the internal K+ concentration is generally somewhat higher than would be expected if the K+ distribution were determined entirely by the membrane potential. Under certain conditions, net uptake of K+ against the electrochemical potential gradient was observed. We suggest that K+ is actively accumulated by a primary transport system that may exchange K+ for H+, and that K+ leaks passively outward through the K+ channel. The K+ circulation across the membrane amounts to about 2% of the K+ pool per min, or 4.5 microA/cm2 of surface area. We propose that this K+ circulation is one arm of the transcellular current, carrying positive charge out of the thallus. PMID- 6281246 TI - Effect of unsaturated fatty acids and Ca2+ on phosphatidylinositol synthesis and breakdown. AB - CDP-diglyceride : inositol transferase was inhibited by unsaturated fatty acids. The inhibitory activity decreased in the following order: arachidonic acid greater than linolenic acid greater than linoleic acid greater than oleic acid greater than or equal to palmitoleic acid. Saturated fatty acids such as myristic acid, palmitic acid, and stearic acid had no effect. Calcium ion also inhibited the activity of CDP-diglyceride : inositol transferase. In rat hepatocytes, arachidonic acid inhibited 32P incorporation into phosphatidylinositol and phosphatidic acid without any significant effect on 32P incorporation into phosphatidylcholine, phosphatidylethanolamine and phosphatidylserine. Ca2+ ionophore A23187 also inhibited 32P incorporation into phosphatidylinositol. However, 32P incorporation into phosphatidic acid was stimulated with Ca2+ ionophore A23187. Phosphatidylinositol-specific phospholipase C was activated by unsaturated fatty acids. Polyunsaturated fatty acids such as arachidonic acid and linolenic acid had a stronger effect than di- and monounsaturated fatty acids. Saturated fatty acids had no effect on the phospholipase C activity. The phospholipase C required Ca2+ for activity. Arachidonic acid and Ca2+ had synergistic effects. These results suggest the reciprocal regulation of phosphatidylinositol synthesis and breakdown by unsaturated fatty acids and Ca2+. PMID- 6281247 TI - Comparison of glycogen phosphorylase kinases of various rat tissues. AB - Glycogen phosphorylase kinases in soluble fractions of various rat tissues were examined for the pH 6.8/8.5 activity ratio, Ca2+-dependency, activation by cyclic AMP-dependent protein kinase (protein kinase A), and reactivity with anti skeletal muscle phosphorylase kinase serum. The enzymes could be divided into at least two major groups; muscle and liver types. The muscle type, that has a low value of pH 6.8/8.5 activity ratio, is highly dependent on Ca2+, markedly activated by protein kinase A, and strongly inhibited by the antiserum. Inversely, the liver type, that has a high value of pH 6.8/8.5 activity ratio, is poorly dependent on Ca2+, not activated by protein kinase A, and weakly inhibited by the antiserum. The enzymes from heart and skeletal muscle were similar and belonged to the former entity. Whereas, the enzymes from liver, kidney, spleen, lung, and testis appeared to belong to the latter entity. The enzyme from brain apparently differs from these entities, and seems to be an intermediate type or a hybrid of the two. PMID- 6281248 TI - A flash-photometric method for determination of reactivity of superoxide: application to superoxide dismutase assay. AB - Pulse-generation of O2- by a flash was used to determine the reactivity of O2-, O2- was produced within 10 ms by a flash of light through the excitation of FMN in the presence of N,N,N',N'-tetramethylethylenediamine and oxygen. Kinetic analysis of cytochrome c reduction by O2- generated by flash yielded the reaction rate constant between cytochrome c and O2- and the spontaneous disproportionation rate constant of O2-. We applied it for superoxide dismutase assay using a linear relation between superoxide dismutase concentration and the apparent rate constant of cytochrome c reduction by O2-. The catalytic rate constant and activation energy at pH 7.3 of bovine liver Cu,Zn-superoxide dismutase were found to be 1.75 x 10(9) M-1 . s-1 at 25 degrees C and 26.9 kJ . M-1, respectively. The kinetics of O2- decay can be also monitored at 240 nm in this flash-photometric system and gave the spontaneous disproportionation rate constant of O2- and the catalytic rate constant of superoxide dismutase. PMID- 6281249 TI - Comparison of properties of the cellular and extracellular phosphodiesterases induced by cyclic adenosine 3',5'-monophosphate in Dictyostelium discoideum. AB - The kinetic properties and susceptibilities to various agents of intracellular (particulate and soluble) and extracellular phosphodiesterases [EC 3.1.4.17] of Dictyostelium discoideum induced by cyclic adenosine 3',5'-monophosphate (cyclic AMP) were studied and compared. Intracellular particulate phosphodiesterase was obtained by solubilization of the light mitochondrial fraction with Emulgen. The Michaelis constants of this enzyme were 4.5 +/- 0.7 and 10 +/- 0.7 microM, while those of the intracellular soluble phosphodiesterase were 4.6 +/- 0.3 and 13 +/- 2.8 microM. However, the Michaelis constant of the extracellular phosphodiesterase was 6.8 +/- 0.9 microM, differing from the values of the two intracellular enzymes. Susceptibilities of the enzyme activity to various agents (theophylline, caffeine, dithiothreitol, glutathione, etc.) were essentially the same among these three phosphodiesterases. In the presence of 10 mM ethylenediaminetetraacetate, the activities of the particulate and the soluble enzymes were both decreased to about 60%, while that of the extracellular enzyme remained at 90%. The inhibition constants of cyclic inosine monophosphate for the cellular enzymes (35 and 100 microM for the particulate enzyme, and 37 and 90 microM for the soluble one) were considerably different from the value for the extracellular enzyme (48 microM). These results suggest that the characteristics of these three phosphodiesterases are substantially similar, but that the affinity of the intracellular (particulate and soluble) enzymes for the substrate is somewhat different from that of the extracellular enzyme. PMID- 6281250 TI - Is transformation associated with an increased error frequency in mammalian cells? AB - Acute starvation of mammalian cells for amino acids results in translational errors that may be detected by two-dimensional polyacrylamide gel electrophoresis. Using this as an assay for error frequency in mammalian cells, we investigated the hypothesis that neoplastic transformation was associated with an increased error frequency which in turn leads to an increased mutation rate and a decreased efficiency of regulatory controls (phenomena of tumor progression). Although we found that transformation was not always associated with an increased level of mistranslation we showed that SV40 transformation increased the level of translational errors in all cell types tested. PMID- 6281251 TI - Myeloperoxidase precursors in human myeloid leukemia HL-60 cells. AB - The biosynthesis of myeloperoxidase was studied in human myeloid leukemia HL-60 cells. Cells were pulse-labeled with [35S]methionine and cell extracts were treated with anti-myeloperoxidase serum. The immunoprecipitates were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. In a 1-h pulse, only two immunoreactive polypeptides of Mr 89,000 and 81,000 were labeled. During subsequent chase, the amounts of these polypeptides decreased with concomitant appearance of polypeptides of Mr 59,500 and 14,700, corresponding to the two chains of myeloperoxidase. Immunoprecipitation of all these polypeptides was prevented by adding excess myeloperoxidase. These results suggest that the Mr 89,000 and 81,000 polypeptides are precursors of myeloperoxidase. The precursors were present only in the soluble fraction of the cells, but the matured polypeptide chains were found in both the soluble and granule fraction after 17-h chase. Only one of the precursors, the Mr 89,000 peptide, was secreted into the culture medium. PMID- 6281252 TI - Recycling of the hepatocyte asialoglycoprotein receptor does not require delivery of ligand to lysosomes. AB - The hepatocyte asialoglycoprotein receptor is able to mediate the internalization of ligand in buffer devoid of Na+ but containing 0.15 M K+. Under these conditions, degradation of internalized ligand does not occur due to an inability to deliver the ligand to lysosomes. Instead, the ligand becomes localized in a vesicle with the same density as plasma membrane on Percoll gradients. This vesicle may be the functional equivalent of the uncoated vesicles observed by electron microscopy. Internalization of more than 20 glycoprotein molecules/high affinity surface receptor was observed under these conditions, indicating that delivery of ligand to lysosomes is not necessary for receptor reutilization. PMID- 6281253 TI - Gonadotropin receptor binding regulators in serum. Characterization and separation of follitropin binding inhibitor and lutropin binding stimulator. AB - Subfractions of human serum capable of inhibiting specific binding of radioiodinated human follitropin and stimulating specific binding of radioiodinated human lutropin to testicular receptors were prepared by molecular sieving techniques, separated by ion exchange chromatography, and further purified by reversed phase high pressure liquid chromatography. Although purified approximately 2000-fold, the exact chemical nature of the follicle-stimulating hormone binding inhibitor is as yet uncertain. However, a variety of analytical procedures indicated it to be separate and distinct from the lutropin binding stimulator, to have a molecular weight of about 700, and to contain a peptide component. Available evidence suggests that the serum factor responsible for luteinizing hormone binding stimulation may be calcium. PMID- 6281255 TI - The cytochrome c peroxidase.cytochrome c electron transfer complex. Experimental support of a hypothetical model. AB - A hypothetical model of the cytochrome c peroxidase.cytochrome c complex (Poulos, T. L., and Kraut, J. (1980) J. Biol. Chem. 255, 10322-10330) predicts charge interactions between aspartic acid residues of the peroxidase having a spatial distribution that is complementary to the distribution of essential and highly conserved residues of cytochrome c. In a first attempt to test this model, carboxyl groups of cytochrome c peroxidase have been modified with a water soluble carbodiimide, either alone or in combination with a nucleophile. Modification led to the loss of up to 90% of the ferrocytochrome c peroxidase activity. At least 4-5 carboxyl groups out of a total of 48, but none of the heme carboxyls, were modified in a derivative with 14% residual activity. In the peroxidase.cytochrome c complex the rate of peroxidase inactivation is slowed and approximately 2 carboxyl groups are protected from chemical modification. In the presence of the carbodiimide, cytochrome c and peroxidase were cross-linked to form a covalent 1:1 complex and the linkage sites were preliminarily characterized. Cross-linking occurred to carboxyl groups of the NH2-terminal fragment 1-119 and of fragment 172-229. The four crucial aspartates of the hypothetical model are located in these same two sequence regions. PMID- 6281254 TI - Phosphorylation of histones 1 and 3 and nonhistone high mobility group 14 by an endogenous kinase in HeLa metaphase chromosomes. AB - We have developed a simple in vitro system for studying phosphorylation in isolated HeLa metaphase chromosomes which utilizes the endogenous protein kinase and phosphoprotein phosphatase activities in the chromosomes. Because the isolated chromosomes retain the specificity of phosphorylation seen in vivo, this system offers unique possibilities for studying the properties and regulation of the kinase and phosphatase by adding exogenous substances and observing their effects. It should also be useful for studying the sites of phosphorylation, since proteins can be more easily labeled to high specific activity with 32P in this system than in vivo. The pattern of proteins phosphorylated in isolated metaphase chromosomes appears to be nearly identical with the pattern found in vivo. Among the histones (H) only H1 and H3 are phosphorylated, but several nonhistone proteins, including high mobility group (HMG) 14, are also phosphorylated. Since HMG 14 has been implicated as a structural protein of actively transcribing chromatin, our results suggest that phosphorylation of chromatin proteins may be involved in the shutoff of transcription during mitosis. Tryptic peptide maps and analysis of the phosphorylated amino acids indicate that H1A, H1B, HMG 14, and H3 are phosphorylated at the same sites in vitro in metaphase chromosomes as in mitotic cells in vivo. The major site of phosphorylation of histone H3, both in vivo and in vitro, has been identified as serine 10. HMG 14 is phosphorylated both at serine and threonine residues. PMID- 6281257 TI - Poly(ADP-ribose) synthetase. The DNA binding domain and the automodification domain. AB - Poly(ADP-ribose) synthetase is known to require double-stranded DNA for activity and to be susceptible to automodification with poly(ADP-ribose). This enzyme is cleaved into two fragments by limited proteolysis with papain. Their molecular weights are 74,000 and 46,000, respectively, as judged by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The fragment of Mr 46,000 binds to a nicked DNA-cellulose column with the same affinity as that of native enzyme, while the fragment of Mr 74,000 has little affinity for the DNA ligand. When the enzyme previously automodified with [adenine-U-14C]NAD (2 ADP-ribose units incorporated per molecule of enzyme) is digested with papain, the label is detected only in the fragment of Mr 74,000. These results indicate that poly(ADP-ribose) synthetase contains two domains, one acting as the site for binding of DNA and the other acting as the site(s) for accepting poly(ADP-ribose). PMID- 6281258 TI - A novel leukotriene produced by stimulation of leukocytes with formylmethionylleucylphenylalanine. AB - Stimulation of human polymorphonuclear leukocytes with the chemotactic peptide formylmethionylleucylphenylalanine led to the formation of a novel leukotriene: 5(S),12(R)-dihydroxy-6,8,10,14-eicosatetraen-1,20-dioic acid. This dihydroxydicarboxylic acid is derived from omega-oxidation of 5(S),12(R),dihydroxy-6,8,10,14-eicosatetradienoic acid (leukotriene B4). The intermediate 5(S),12(R),20-trihydroxy-6,8,10,14-eicosatetraenoic acid was also isolated from these incubations. The two metabolites of leukotriene B4 exhibit chemotactic properties for human polymorphonuclear leukocytes but are less active in this respect than the parent compound. PMID- 6281256 TI - Protection by superoxide dismutase, catalase, and poly(ADP-ribose) synthetase inhibitors against alloxan- and streptozotocin-induced islet DNA strand breaks and against the inhibition of proinsulin synthesis. AB - We have shown previously that alloxan and streptozotocin, two major diabetogenic agents, cause DNA strand breaks in rat pancreatic islets and stimulate nuclear poly(ADP-ribose) synthetase, thereby depleting intracellular NAD level and inhibiting proinsulin synthesis (Okamoto, H. (1981) Mol. Cell. Biochem. 37, 43 61; Yamamoto, H., Uchigata, Y., and Okamoto, H. (1981) Nature 294, 284-286). In the present study, superoxide dismutase and catalase, scavengers of radical oxygens, were found to protect against islet DNA strand breaks and inhibition of proinsulin synthesis induced by alloxan. The radical scavengers did not affect islet DNA strand breaks or inhibition of proinsulin synthesis induced by streptozotocin. On the other hand, compounds that inhibit islet nuclear poly(ADP ribose) synthetase were found to protect against alloxan- as well as streptozotocin-induced inhibition of proinsulin synthesis. The poly(ADP-ribose) synthetase inhibitors were ineffective in protection against DNA strand breaks induced by the agents. These results may provide an important clue for elucidating the prevention of insulin-dependent diabetes as well as for understanding the cause of diabetes. PMID- 6281259 TI - Regulation of myosin light chain and phosphorylase phosphorylation in tracheal smooth muscle. PMID- 6281260 TI - Nucleic binding affinity of bacteriophage T4 gene 32 protein in the cooperative binding mode. AB - This study reports on various parameters which affect the binding stoichiometry for complexes of bacteriophage T4 gene 32 protein (P32) and single stranded polynucleotides (determined by UV absorbance and fluorescence quenching) and presents results of a quantitative electron spin resonance assay to determine physiologically effective binding affinity differences of nucleic acid binding proteins. The assay employs macromolecular spin probes (spin-labeled nucleic acids) which are used to determine the fraction of saturation in competition experiments with unlabeled nucleic acids. It was found that the fraction of complexed spin-labeled polynucleotides can be directly monitored by ESR with a two-component analysis approach when ligands such as poly(L-lysine), gene 5 protein (P5) of filamentous bacteriophage fd, and gene 32 protein (P32) of bacteriophage T4 are used. The ESR data unequivocally show that: 1) the binding stoichiometry for poly(L-lysine), P5 and P32 is nucleotide/lysine, 4 nucleotides/P5 monomer, and 10 nucleotides/P32 monomer, respectively; and 2) under physiologically relevant buffer conditions the relative affinity of P32 in the cooperative binding mode for polythymidylic acid is about 4 times greater than for polydeoxyinosinic acid and about 12 times greater than for polyinosinic acid, and the relative affinity of P32 for polydeoxyinosinic acid is about 3 times greater than for polyinosinic acid. PMID- 6281261 TI - Kinetic characterization of the interaction between cytochrome oxidase and cytochrome c. AB - The mechanism of electron transfer catalyzed by cytochrome oxidase was investigated by monitoring the reaction of cytochrome oxidase with cytochrome c under carefully controlled anaerobic conditions. The kinetics of the reaction were examined by varying conditions of ionic strength, inhibitor binding, and oxidation-reduction potential. An analogue of cytochrome c in which the iron atom was replaced with cobalt was used to probe the effect of redox potential on the reaction. Under conditions of low ionic strength, there is very rapid oxidation of cytochrome c and reduction of oxidase which occurs at a rate of 3 X 10(7) M-1 s-1. The number of electrons transferred exhibit a hyperbolic dependence on the concentration of cytochrome c reaching a maximum of 2 electrons transferred at the highest concentration of reduced cytochrome c employed. The total number of electrons transferred was always observed to be distributed equally between cytochrome a and a second acceptor which appears to be the associated copper center; electron transfer to cytochrome a3 did not occur in the absence of oxygen. Substitution of cytochrome c by the cobalt analogue (which represents a decrease in oxidation-reduction potential of about 400 mV) yielded identical results indicating that the origin of the lack of reactivity of cytochrome a3 is of a kinetic nature. The effect of increasing the ionic strength on the reaction was 2-fold: a marked decrease in reaction rate and the appearance of biphasic kinetics with the amplitude of the very fast absorbance changes at 605 nm decreasing from 80% to 40% of the total anticipated from static absorbance measurements. Each of the two phases accounted for a maximum of 1 electron at the highest ionic strength employed. These results are simulated in terms of a sample kinetic reaction scheme involving a two-step electron transfer at one binding site. PMID- 6281262 TI - Protein n, a primosomal DNA replication protein of Escherichia coli. Purification and characterization. AB - Protein n, essential in forming the primosome for the in vitro conversion of phi X174 single-stranded (SS) DNA to the duplex replicative form (RF), has been purified about 5000-fold to near homogeneity from Escherichia coli. Protein n is heat- and acid-resistant and N-ethyl-maleimide-sensitive. It appears to be a dimer of 12,000 (+/- 2000)-dalton polypeptides. About 80 molecules of protein n are present/cell. Protein n binding to phi X SS DNA depends on the presence of single-strand binding protein (SSB). This requirement for SSB reflects a direct interaction of protein n and SSB. About 30 protein n monomers can be bound to an SSB-coated circle. However, in forming the primosome on an SSB-coated phi X circle, an input of only 2-3 protein n monomers is required and 1 monomer bound/circle. Retention of this low level of protein n on SSB-coated phi X SS DNA is dependent upon protein n', a DNA-dependent ATPase (dATPase) that guides primosome assembly. This single protein n monomer is retained in the assembled primosome, which is conserved on the completed parental RF and participates in the next stage of the replicative cycle, production of progeny RF. PMID- 6281263 TI - Interconversions of [3Fe-3S] and [4Fe-4S] clusters. Mossbauer and electron paramagnetic resonance studies of Desulfovibrio gigas ferredoxin II. PMID- 6281264 TI - Adaptation of human neutrophil responsiveness to the chemoattractant N formylmethionylleucylphenylalanine. Heterogeneity and/or negative cooperative interaction of receptors. AB - The dose-response characteristics of the neutrophil 3-3'-dipentyloxacarbocyanine (di-O-C5(3)) fluorescence response to repetitive stimulation with the chemoattractant N-formylmethionylleucylphenylalanine (fMet-Leu-Phe) were studied. Neutrophils exposed to the chemoattractant fMet-Leu-Phe at less than 5 X 10(-8) M subsequently responded only to higher concentrations of fMet-Leu-Phe. This stimulus-induced modification of neutrophil responsiveness involved a reversible fMet-Leu-Phe-induced shift in response Km (the concentration of fMet-Leu-Phe producing a half-maximal response) to higher values which occurred 1 to 2 min after exposure to fMet-Leu-Phe and represented a form of adaptation. A Hill coefficient of 0.68 +/- 0.07 was determined from analysis of the data indicating that the di-O-C5(3) fluorescence response behavior is compatible with functional negatively cooperative interaction and/or heterogeneity of fMet-Leu-Phe receptors. In related studies, analysis of the binding of fMet-Leu-[3H]Phe to intact cells and cell-free plasma membrane preparations resulted in Hill coefficients of 0.64 +/- 0.06 and 0.69 +/- 0.07, respectively, indicating that fMet-Leu-Phe binding exhibits properties similar to the fMet-Leu-Phe-elicited di O-C5(3) fluorescence response. Modulation of receptor affinity, through either negative cooperativity or changing populations of heterogeneous receptors, may be an important mechanism by which neutrophils adapt and respond to a gradient of chemoattractant during the process of chemotaxis. PMID- 6281265 TI - Hormone-sensitive adenylate cyclase. Delineation of a trypsin-sensitive site in the pathway of receptor-mediated inhibition. PMID- 6281266 TI - Relationship between superhelical density and cruciform formation in plasmid pVH51. AB - The relative stability of the cruciform state at the large inverted repeat of plasmid pVH51 is measured. At physiological superhelical densities, the cruciform state is present in a high percentage of the plasmid molecules. Investigation of the relationship between negative superhelical density and cruciform prevalence reveals a sharp transition from an undetectable level to a relatively stable state. This transition occurs over the negative superhelical density range of 0.046 to 0.066. Estimates of the free energy contribution to cruciform formation resulting from loss of negative superhelical turns suggest that about 22 kcal/mol are required to generate the cruciform structure at this site in pVH51. PMID- 6281267 TI - The gastric [H,K]ATPase:H+/ATP stoichiometry. AB - An H+/ATP ratio of 2 for H+ transport was determined from initial rate measurements at pH 6.1 in a purified gastric microsomal fraction containing the [H,K]ATPase. This ratio was independent of external KCl, though the apparent K0.5 for ATP was increased from 10.78 +/- 0.51 (n = 3) to 64.6 +/- 11.9 (n = 3) microM ATP and from 5.13 +/- 0.64 (n = 3) to 65.2 +/- 0.64 (n = 3) microM ATP for H+ transport and the K+-stimulated ATPase, respectively, as K+external was increased from 12 to 150 mM. The H+/ATP ratio was also relatively independent of ATP concentration. Maximum initial rates obtained in KCl-equilibrated vesicles were independent of added valinomycin, though net H+ transport was increased 29.3 +/- 1.03% (n = 6) by the addition of ionophore. Maximum net H+ transport in this vesicle preparation was 185 +/- 2.1 (n = 14) nmol mg-1 of protein. Initial rate measurements of ATPase represent a burst of K+-dependent activity of approximately 10-15 s duration. The H+/ATP stoichiometry was calculated based on the K+-stimulated component of hydrolysis. Under most conditions, the Mg2+ dependent component of hydrolysis was less than 10% of the (Mg2+ + K+) component of hydrolysis. PMID- 6281268 TI - Covalent affinity labeling by periodate-oxidized [alpha-32P]ATP of the large-T proteins of polyoma and SV40 viruses. AB - Incubation of crude extracts from cells lytically infected with polyoma virus in the presence of periodate-oxidized [alpha-32P]ATP led to the radioactive labeling of one main polypeptide immunoprecipitated by anti-T antigen antibodies. It was absent from extracts of mock-infected cells and exhibited an apparent Mr value of 105,000, identical with that of the large-T viral protein. This polypeptide was unambiguously identified as large-T on the basis of the heat sensitivity of the in vitro labeling in extracts from cells infected with a tsa mutant. The amount of incorporated radioactivity was found to increase linearly with that of infected cell extract and with the incubation time. Labeling resulted from the specific binding of an oxidized ATP molecule to a nucleotide binding site of the large-T protein, since it was efficiently completed by unlabeled ATP. Study of the dependence on the concentration of oxidized ATP evidenced a first order kinetic for the labeling reaction. Similar results were obtained for the large-T protein of SV40 virus. PMID- 6281269 TI - Escherichia coli sulfite reductase hemoprotein subunit. Prosthetic groups, catalytic parameters, and ligand complexes. AB - Escherichia coli NADPH-sulfite reductase can be dissociated into an oligomeric flavoprotein and a monomeric hemoprotein (HP) subunit in 4 M urea. HP catalyzes stoichiometric 6-electron reductions of SO32- (to S2-) and of NO2-, as well as 2 electron reduction of NH2OH, with reduced methyl viologen (MV+) as reductant. While Vmax values are highest with the nitrogenous substrates, Km for SO32- is 2 to 3 orders of magnitude less than the Km for NO2- or NH2OH. EPR spectroscopic and chemical analyses show that HP contains one siroheme and one Fe4S4 center per polypeptide. The heme is in the high spin Fe3+ state in HP as isolated. Near quantitative reduction of the Fe4S4 center to a state yielding a g = 1.94 type of EPR spectrum by S2O42- and/or MV+ could be achieved if HP was converted to either the CN- or CO complex or treated with 80% dimethyl sulfoxide. HP binds one SO32- or CN- per peptide. Binding of these ligands, as well as CO, appears to be mutually exclusive and to involve the heme. The heme Fe3+/Fe2+ potential is shifted from -340 mV in the free HP to -155 mV in the HP-CN- complex. The potential of the Fe4S4 center is approximately 70 mV more negative in the CN- as opposed to the CO-ligated HP (-420 mV), a result which indicates the presence of heme-Fe4S4-ligand interaction in the HP complexes. PMID- 6281270 TI - Effects of mutational loss of adenosine kinase and deoxycytidine kinase on deoxyATP accumulation and deoxyadenosine toxicity in cultured CEM human T lymphoblastoid cells. PMID- 6281271 TI - Lipid hapten containing membrane targets can trigger specific immunoglobulin E dependent degranulation of rat basophil leukemia cells. AB - We have studied the binding of liposomes containing dinitrophenylated lipid to rat basophil leukemia cells armed with monoclonal anti-dinitrophenyl IgE. The liposomes were either "fluid" at 37 degrees C (dimyristoylphosphatidylcholine or an equimolar binary mixture dipalmitoylphosphatidylcholine and cholesterol) or "solid" (dipalmitoylphosphatidylcholine, distearoylphosphatidylcholine, or dibehanoylphosphatidylcholine). We have also studied the immune mediated degranulation of these cells induced by the above lipid membrane targets. In some cases both studies were carried out with liposomes containing various surface densities of lipid haptens. From these studies we conclude that freely mobile nonaggregated lipid haptens in bilayer membrane targets can trigger efficient serotonin release from rat basophil leukemia cells in the presence of specific antihapten IgE. Solid target membranes are also effective as stimulators of serotonin release. The release of serotonin depends strongly on the surface density of lipid haptens over a narrow range of surface densities. These studies with lipid membrane targets having well defined physical properties indicate the need for generalized molecular models of receptor-mediated cell triggering. PMID- 6281272 TI - Protein-protein interactions of cytochrome oxidase in inner mitochondrial membranes. The effect of liposome fusion on protein rotational mobility. AB - Rotational diffusion of cytochrome oxidase in the inner membrane of rat liver mitochondria was measured by detecting the decay of absorption anisotropy after photolysis of the heme a3.CO complex by a vertically polarized laser flash. As in previous experiments with beef heart mitochondria (Kawato, S., Sigel, E., Carafoli, E., and Cherry, R. J. (1980) J. Biol. Chem. 255, 5508-5510), co existence of rotating cytochrome oxidase (mean rotational relaxation time, phi, of 700 to 1400 microseconds) and immobilized cytochrome oxidase (phi greater than 20 ms) was observed in mitochondria and mitoplasts. The effect of lipid/protein ratio by weight (L/P) on the relative proportions of mobile and immobile cytochrome oxidase was investigated following the fusion of soybean phospholipid vesicles with mitoplasts. The fusion procedure yielded four separate fractions upon sucrose density gradient centrifugation with L/P as follows: 0.3 in Pellet, 0.7 in Band 3, 1.5 in Band 2, and 3.0 in Band 1. The percentage of rotationally mobile cytochrome oxidase (phi = 700 to 1000 microseconds) in each of the different bands was found to be 16% in Pellet, 25% in Band 3, 47% in Band 2, and 76% in Band 1 at 37 degrees C. The dependence of the amount of mobile cytochrome oxidase on L/P indicates that the fraction of aggregated protein progressively decreases with decreasing concentration of proteins in the membrane. Thus, the large immobile fraction of cytochrome oxidase in mitochondrial inner membranes can be explained by nonspecific protein aggregation which is a consequence of the low L/P. The decrease in the mobile fraction in Pellet compared with mitoplasts was shown to be due to the pH 6.5 incubation used for fusion. PMID- 6281273 TI - Transcriptional initiation and processing of the small ribosomal RNA of yeast mitochondria. AB - We have identified the nucleotide at which transcription initiates on the yeast mitochondrial small (14 S) rRNA gene by sequencing of RNA labeled at the 5' initiating triphosphate with vaccinia virus guanylyltransferase [alpha-32P]GTP (in vitro capping reaction). Initiation occurs within the stem of a 12-base palindromic repeat. The initiation sequence has homology with the large (21 S) ribosomal RNA initiation sequence that has been previously determined. We have also sequenced the 5' and 3' ends of the mature 14 S rRNA after labeling with T4 polynucleotide kinase and RNA ligase, respectively. These sequences demonstrate that about 80 nucleotides are cleaved from the 5' end of a precursor to produce the mature 14 S rRNA. This cleavage is imprecise in that the processing occurs at one of five adjacent nucleotides 77 to 81 nucleotides downstream from the 5' initiation site. The 3' ends of this precursor and the mature 14 S rRNA are unique and identical. PMID- 6281274 TI - Interaction of Bacillus subtilis RNA polymerase core with two specificity determining subunits. Competition between sigma and the SPO1 gene 28 protein. AB - Gene activity in the development of phage SPO1 is transcriptionally regulated. Early viral genes are transcribed by the major vegetative cell RNA polymerase (E. sigma). Middle viral genes are transcribed by RNA polymerase core (E) bearing the SPO1 gene 28 protein (gp28) instead of sigma. This paper deals with the competitive interactions of sigma and gp28 with E which must, at least in part, be involved in the ability of viral middle gene expression to succeed early gene expression. An in vitro assay has been developed for determining the proportions of early (E. sigma) and SPO1 middle (E.gp28) Bacillus subtilis RNA polymerase in mixtures. The assay, which is based on the transcription of two well studied SPO1 restriction fragments followed by gel electrophoresis and quantitation of the E. sigma and E.gp28 transcription products, has been used to study the interactions of sigma and gp28 with RNA polymerase core. These subunits are found to be capable of competitively excluding each other. The binding competition is sensitive to ionic strength, with sigma being more effective below 0.2 total ionic strength and gp28 being more effective at higher values. The outcome of competition is also dependent on the order of addition of subunits and the reconstitution kinetics have been studied. The subunit competition is rather insensitive, particularly at higher ionic strength, to temperature. Gp28 can convert E. sigma to E.gp28 but the reverse reaction occurs much less efficiently. The B. subtilis delta protein biases the sigma-gp28 competition in favor of the sigma subunit. the implications of these results for the physiological transition of transcriptional specificity during SPO1 development are discussed. PMID- 6281275 TI - Purification and characterization of protein IIIb, a mammalian brain phosphoprotein. AB - The phosphorylation of a 55,000-dalton protein (Protein IIIb) present in mammalian brain was previously shown to be increased by depolarizing agents in the presence of calcium, by cyclic nucleotides, and by appropriate neurotransmitters. We now report that Protein IIIb has been purified 660-fold to near homogeneity and partially characterized. The hydrodynamic properties of the purified protein indicate that it exists as an elongated monomer. cAMP-dependent protein kinase catalyzes the incorporation of 0.82 mol of phosphate into serine/mol of protein. The protein is heterogeneous in isoelectric focusing, exhibiting multiple forms with isoelectric points ranging in pH from 6.6 to 7.3. PMID- 6281276 TI - Participation of 5'-terminal leader sequences in in vitro translation of Rous sarcoma virus RNA. AB - The cell-free translation of genome RNA from Rous sarcoma virus was examined following hybridization to selected fragments of viral DNA. Single-stranded fragments, generated by t-RNATrp primed transcription of 70 S RNA from the Schmidt-Ruppin D strain, were isolated and purified by electrophoresis. These included DNA complementary to the 5'-terminal 101 nucleotides (DNA100) of virion 38 S RNA and a collection of prematurely terminated transcripts which lack the complement to the extreme 5'-terminal 7-20 nucleotides (DNA less than 100). In addition, DNA encompassing the viral leader sequences was purified from a cloned copy of proviral DNA. Under hybrid-arrested translation conditions, the leader DNA as well as DNA100 inhibited translation of all protein products generated from the 70 S RNA, while hybridization to the shorter transcripts (DNA less than 100) did not affect in vitro protein synthesis. All single-stranded DNAs were shown to hybridize with equal efficiency to viral RNA under hybrid-arrested translation conditions and inhibition of protein synthesis by DNA100 was concentration-dependent. These results document the participation of noncoding leader RNA in Rous sarcoma virus protein synthesis and demonstrate that a free, single-stranded 5' terminus is necessary for cell free translation of 70 S RNA. PMID- 6281277 TI - Polyoma-like particle: characterization of the DNA encapsidated in vitro by polyoma empty capsids. AB - Polyoma empty capsids and DNA interact in a cell-free system to form a polyoma like particle (PLP). The particle is stable in high concentrations of salt and contains a DNA fragment that is protected by the capsid from the action of DNase I. The DNA extracted from PLP is heterogeneous in size. It has a mean molecular weight of 1.2 x 10(6) with a standard deviation of +/- 0.5 x 10(6). Analysis of PLP DNA with restriction endonucleases and the Southern blot technique reveals that a specific primary sequence is not required for PLP formation. Either linear, circular, or supercoiled polyoma DNA, as well as single-stranded DNA, rRNA, and the synthetic homopolymers poly(dA) . poly(dT) and poly(dG) . poly(dC) can be used for PLP formation. It is concluded that polyoma empty capsids can be used to bind and encapsidate nucleic acids differing in their primary, secondary, and superhelical tertiary structure to form, in vitro, stable polyoma-like particles. PMID- 6281278 TI - Characterization of transcripts from the Var1 region on mitochondrial DNA of Saccharomyces cerevisiae. AB - We have identified transcripts with sequence homology to the var1 region on yeast mtDNA. In wild type, and in cytoplasmic petite strains retaining the var1 region, we detect four RNA species, 19 S, 16 S, 14 S, and 13 S, which hybridize to var1 DNA probes. The 16 S RNA is by far the most abundant of these RNAs in wild type cells. We also observe hybridization of the 15 S rRNA and a 10 S RNA species to var1 DNA probes. This hybridization is most likely due to the presence in these RNAs of a GC-rich cluster which has near perfect sequence homology to a GC-rich cluster in the var1 region. We find that the 16 S RNA, estimated to be 2000 to 2100 nucleotides long, varies in size in direct correspondence with the size of var1 polypeptide; it is about 100 nucleotides longer in strains with the var1 (44,000) allele than in strains with var1 (40,000). The amount of the 16 S RNA also varies in correspondence with the amount of var1 polypeptide made; it is increased in an oxi-3 mit- strain (CAD245) which makes about 10 times more var1 protein than wild type, and is barely detected in PZ200, a strain with very low levels of mitochondrial protein synthesis harboring two mutations within var1. We have purified the 16 S RNA following chromatography over oligo(dT) cellulose columns. When the RNA is end-labeled and hybridized to a HincII digest of wild type mtDNA, we observe hybridization only to the fragment containing the entire var1 region, HincII fragment 10. PMID- 6281279 TI - Effect of deoxynucleoside phosphorothioates incorporated in DNA on cleavage by restriction enzymes. AB - DNA synthesized in vitro using deoxynucleoside phosphorothioates as substrates is quite similar to normal DNA in its biochemical properties (Vosberg, H.P., and Eckstein, F. (1977) Biochemistry 16, 3633-3640). In order to investigate the effect of phosphorothioate groups in DNA on the cleavage pattern of restriction endonucleases phosphorothioate double-stranded, circular, replicative form of fd DNA was synthesized in vitro with Escherichia coli DNA polymerase I using native single-stranded DNA as template and mixtures of three normal nucleotides and one nucleoside phosphorothioate analogue as substrates. The double-stranded products were hybrids with respect to their phosphorothioate content. Restriction analysis of normal and phosphorothioate DNA with the restriction endonucleases Hae III, Bam HI, Hpa II, HindII, Alu I, and Taq I showed that the enzymes were inhibited to different degrees depending on which of the nucleotides was replaced by the phosphorothioate. Most significant, inhibition was seen throughout with those DNAs which contained a phosphorothioate exactly at the cleavage site. Phosphorothioate substitutions at other positions, but still within the recognition sequences, were, except for Alu I, not or weakly inhibitory. Phosphorothioate nucleotides not present in the recognition sequences did not affect at all the fragment patterns. The results show that recognition sequences of restriction endonucleases can be selectively protected against cleavage by base-specific introduction of phosphorothioate groups into DNA. PMID- 6281280 TI - Purification of the TET repressor and TET operator from the transposon Tn10 and characterization of their interaction. PMID- 6281281 TI - Localization of dolichol in the lysosomal fraction of rat liver. AB - The distribution of dolichol and/or dolichol esters in subcellular fractions prepared from a rat liver homogenate has been investigated. After saponification of the various fractions dolichol was isolated and quantitated by high performance liquid chromatography in three systems. The degree of purity of the subcellular preparations was examined by marker enzymes and by electron microscopy. Using differential centrifugation it was found that the level of dolichol was highest in the mitochondria-lysosome fraction. Upon further resolution of this fraction by sucrose density centrifugation it was found that the majority of the dolichol was associated with the lysosome-rich fraction. In contrast, the mitochondrial fraction had only a low level of dolichol. This novel observation was confirmed by the finding that dolichol was greatly enriched in a highly purified lysosome fraction preparations isolated by Metrizamide density centrifugation. The enrichment of dolichol in this purified preparation paralleled the observed enrichment of the lysosomal enzyme activity in this fraction. All of these data suggest that the majority of cellular dolichol and/or dolichol esters is localized in the lysosome fraction. The significance of this finding in relation to the metabolism of dolichol is discussed. PMID- 6281282 TI - gamma-Aminobutyric acid and postganglionic sympathetic transmission in the pulmonary artery of the rabbit. AB - 1 The effect of gamma-aminobutyric acid (GABA) on postganglionic sympathetic neurotransmission was studied in strips of the rabbit pulmonary artery. The strips were preincubated with 3H-noradrenaline and then superfused with 3H-amine free medium. They were stimulated either electrically at 2 Hz, or by 60 mM potassium, or by 1 microM tyramine. 2 GABA (1 - 1000 microM) did not change the basal outflow of tritium, but decreased the electrically evoked overflow as well as the contractile response. GABA 1 microM decreased the evoked overflow by 12%, and GABA 1000 microM, by 42%. The effect of GABA was not changed by yohimbine, propranolol, cocaine, corticosterone, or indomethacin. It was not antagonized by picrotoxin or bicuculline methiodide. GABA 100 microM also slightly reduced the potassium-evoked overflow of tritium but did not change the tyramine-evoked overflow. 3 The results show that, in the pulmonary artery of the rabbit, GABA inhibits the release of noradrenaline. Its effect is independent of alpha- and beta-adrenoreceptors and is not mediated by prostaglandins. The effect may be due to activation of presynaptic receptors which appear to differ from conventional GABA receptors inasmuch as they are insensitive to blockade by either picrotoxin or bicuculline. PMID- 6281283 TI - Induction of differentiation in human promyelocytic leukemia cells by tumor promoters. AB - 12-)-Tetradecanoylphorbol-13-acetate (TPA), the prototype polyfunctional diterpene ester tumor promoter of two-step carcinogenesis in mouse skin, induced differentiation of human promyelocytic leukemia cells (HL-60) in culture. Differentiation of HL-60 cells was characterized by increased phagocytosis, increased lysozyme activity (EC 3.2.1.17) in the growth medium, and changes in morphology to those characteristics of more mature cells resembling macrophages. Many of the cells treated with TPA became aggregated, attaching firmly to culture flasks. The average intracellular myeloperoxidase activity (EC 1.11.1.7) per cell decreased during induction of differentiation by TPA. It was also found that TPA enhanced, rather than inhibited, differentiation of HL-60 cells induced by DMSO. In addition to TPA, several polyfunctional diterpene esters of the tigliane, ingenane, and daphnane type have been tested for their ability to induce morphological and functional changes of HL-60 cells. The activities of the compounds to induce these changes correlated well with their activities as tumor promoters in two-step carcinogenesis in mouse skin. In particular, half the concentrations required for induction of adhesion of the cells to flasks were roughly correlated to the potency of these compounds as tumor promoters. Among the compounds tested, phorbol-12,13-didecanoate (PDD), ingenol-3-hexadecanoate, Pimelea factor P1 and Pimelea factor P2 were as active as TPA, while 4-O-methyl TPA and 4 alpha-PDD were much less active. Phorbol and ingenol were totally inactive up to a concentrations 10,000-fold higher than that of TPA. PMID- 6281285 TI - Determination of cefsulodin, cefotiam, cephalexin, cefotaxime, desacetyl cefotaxime, cefuroxime and cefroxadin in plasma and urine by high-performance liquid chromatography. AB - Closely related methods for the determination of several cephalosporins in plasma and urine are described. Deproteinized plasma or diluted urine is directly injected on a RP-8 or RP-18 bonded-material column. Chromatography is performed either in the reversed-phase or the ion-pair mode. The limits of sensitivity range from 0.4 to 2 mumol of cephalosporins per liter of plasma, and from 20 to 100 mumol per liter of urine. The sensitivity may be improved two to five times by using precolumn loading, direct sample clean-up and automatic injection. The stability of the cephalosporins in plasma, urine and water and the reproducibility and accuracy of the methods are reported. PMID- 6281286 TI - Normal-phase high-performance liquid chromatography of porphyrin free acids on silica modified with tetraethylenepentamine. PMID- 6281284 TI - Role of natural prostaglandins in the control of murine mammary tumor virus expression. AB - The regulation of murine mammary tumor virus (MuMTV) production by mammotropic hormones, hormonomimetic substances, and cyclic nucleotides was investigated. The virus produced in control and treated mammary tumor cell cultures was quantitated by measuring the supernatant reverse transcriptase activity in exogenous reaction using poly(rC).oligo(dG) as template-primer. Two days after exposure, the synthetic glucocorticoid, dexamethasone (DXMT), increased spontaneous MuMTV production at optimal concentration (0.1 mumol) up to ten times. Dibutyryl derivative of cyclic AMP had no effect on spontaneous MuMTV production, whereas the drug potentiated suboptimal concentrations of the glucocorticoid. Natural prostaglandins, potent agonists of adenylate cyclase catalyzing intracellular synthesis of cyclic AMP, enhanced both basal (up to five times) and DXMT stimulated (up to 1.6 times) MuMTV replication. The MuMTV-stimulating activity of prostaglandins decreased in the order of PGA1 greater than PGE1 greater than PGB1 greater than PGF2 alpha. Prostaglandins can be replaced partially by norepinephrine and isoproterenol by enhancing the DXMT-mediated MuMTV stimulation, whereas these drugs remained without effect on spontaneous MuMTV production. Theophylline, an antagonist of cAMP-phosphodiesterase converting cAMP to AMP, enhanced the virus-stimulating activity of DXMT as well as of prostaglandins. The enhancement of MuMTV production by adenylate cyclase agonists do not correlate absolutely with the estimates of intracellular cAMP levels, since the highest amounts of cAMP has been repeatedly observed in cells treated with PGE1 and norepinephrine. The results indicate that besides hormones, other hormone-like substances and cyclic nucleotides may be involved in the complex mechanism of hormone-regulated MuMTV genome expression. PMID- 6281287 TI - Human rotavirus RNA prepared from stool samples by a simple procedure suitable for the determination of electropherotypes. PMID- 6281289 TI - Evaluation of a cross-flow filtration technique for extraction of polioviruses from inoculated oyster tissue homogenates. AB - Pleated membrane filtration capsules, utilizing cross-flow and liquid membrane technology, were evaluated for use in poliovirus extraction from inoculated oyster tissue homogenates. The filtration technique is fast and less complex than other extraction procedures. Capsules filtered an average of 450.3 ml of a one liter homogenate within 20 min. Filtrates were clear and contained a mean virus recovery of 46.1% in non-dialyzed samples that were concentrated by ultrafiltration. Cytotoxicity was not observed in cell culture monolayers inoculated with non-dialyzed, water-dialyzed, or buffer-dialyzed concentrates. Dialysis and concentration procedures did not significantly reduce the number of viruses in the filtrates. PMID- 6281288 TI - A reproducible method for the optimum infection of human mononuclear cells and lymphoid cell lines by herpes simplex virus type 1. AB - Parameters for the infection of human mononuclear cells (MNC) with herpes simplex virus type 1 (HSV-1) were investigated and a procedure was established which resulted in a reproducible optimum number of cells expressing virus-specific cell surface antigens. The number of cells infected was independent of sex of the donor and independent of whether the donor was HSV-seropositive or -seronegative. On the average 18 +/- 6% of HSV-infected MNC from any given donor expressed HSV specific cell surface antigens. When the standard procedure was applied to a variety of lymphoid cell lines, a high percentage of cells of both B and non T/non-B lines expressed HSV-specific cell surface antigens, whereas T-cell lines appeared resistant to HSV infection. PMID- 6281290 TI - Evaluation of solubilized herpes simplex virus membrane antigens in diffusion in gel-enzyme-linked immunosorbent assay (DIG-ELISA). AB - Cell membranes were prepared from herpes simplex virus (HSV) type 1-infected cells and solubilized with a low concentration of sodium deoxycholate. The supernatant after ultracentrifugation was used as antigen in a newly developed solid phase assay, diffusion in gel-enzyme-linked immunosorbent assay (DIG ELISA). Antigen solubilization was almost complete, all HSV glycoproteins were represented, the yield of antigen in the solubilization process was high, and the presence of a detergent, sodium deoxycholate, did not interfere with the adsorption of antigen to the solid phase. DIG-ELISA was compared with the neutralization test for the determination of HSC antibodies and zone diameters showed a good correlation with the titre obtained by the neutralization test. PMID- 6281291 TI - Hepatocellular cancer in Sweden: incidence 1961-1962 and 1971-1972. AB - The present study was undertaken to compare the incidence of hepatocellular cancer in Sweden during 1961-1962 and 1971-1972, and to evaluate the geographical distribution of the disease. Another aim was to determine whether a retrospective analysis of hospital records of individual cases could provide information on relevant risk factors, in particular cirrhosis, previous diseases, occupational exposure and alcohol consumption. The incidence of hepatocellular cancer in Sweden has increased, particularly in the highest age groups. A difference in incidence for males between the three largest cities in Sweden and the remainder of the country is demonstrated. The proportion of autopsies in Sweden, particularly in the highest age groups, has increased between the two time periods, which could explain part of the increase in incidence. Information from the hospital records indicate that cirrhosis and alcohol abuse among the male cases in the three largest cities are more common than in the remainder of Sweden. PMID- 6281292 TI - Effect of thyroid status on ouabain binding to the human lymphocyte. AB - Lymphocyte Na-K ATPase was evaluated as an index of thyroid status in man. Lymphocytes from 24 untreated hypothyroid patients and 11 hyperthyroid subjects were sampled in parallel with normal lymphocytes, and Na-K ATPase activity was assessed by measurements of ouabain binding to a plasma membrane fraction or to whole cells. In both systems, ouabain bound saturably and specifically, resulting in linear Scatchard plots. Normal lymphocyte plasma membranes bound 2.30 +/- 0.16 pmol ouabain/mg protein (mean +/- SEM; n = 11), with a Kd of 68 +/- 12 nM. Intact normal lymphocytes bound 3.24 +/- 0.30 pmol ouabain/10(7) cells (n = 14), representing 189,000 sites/cell. In hypothyroidism, ouabain binding, when compared with normal cells sampled on the same day, was reduced by 22.0 +/- 5.3% (n = 11; P less than 0.001) in plasma membranes and by 29.1 +/- 3.5% (n = 14) in whole lymphocytes (P less than 0.001), but there was no significant change in the Kd in the membrane fraction. In 6 subjects, the decrease in ouabain binding to lymphocytes was reversed by thyroid hormone replacement. Red cells from hypothyroid subjects showed normal ouabain binding. Ouabain binding to hyperthyroid plasma membranes (2.42 +/- 0.18 pmol/mg protein) was not significantly different from normal. The results in hypothyroid subjects are consistent with the hypothesis that lymphocyte Na-K ATPase is regulated by thyroid hormones. However, lymphocyte Na-K ATPase does not increase in parallel with elevated thyroid hormone levels in hyperthyroidism. The mechanisms underlying these observations remain to be clarified. PMID- 6281293 TI - Escape from mineralocorticoid excess: the role of angiotensin II. AB - Escape from the sodium-retaining action of mineralocorticoids coincides with the suppression of plasma renin and angiotensin II levels. The purpose of this study was to evaluate whether blockade of the renin system accelerates this escape. Eight male normotensive volunteers, aged 24--33 yr, were maintained during two subsequent periods of 12 days each, separated by 3--4 weeks, on a constant intake of sodium and potassium of 140 mmol/day. During both periods, fludrocortisone acetate (0.2 mg) was administered orally three times a day on days 4--12. In addition, on days 3--12, either a converting enzyme inhibitor (MK 421;20 mg orally, twice daily) or a placebo was added in double blind fashion and randomized sequence. During both periods, blood pressures were similar; they tended to increase slightly toward day 12. The weight increase did not differ between the two periods. With MK 421, angiotension II levels were significantly lower than with placebo on days 3--6 (P less than 0.001). On the same days, PRA was increased due to converting enzyme blockade. Despite the significantly different angiotensin II levels on days 3--6, daily urinary sodium excretion on all individual days as well as cumulative sodium balance were the same during both periods. Therefore, we could find no evidence in man that suppression of circulating angiotensin II levels is causally related to escape from mineralocorticoid excess. PMID- 6281294 TI - Cimetidine treatment of azotemic secondary hyperparathyroidism. AB - Cimetidine, an antagonist to histamine H2-receptors, reportedly lowers serum calcium and/or serum immunoreactive parathyroid hormone (iPTH) concentrations in some patients with primary and secondary (azotemic) hyperparathyroidism. We administered the drug orally (300 mg every 6 h) to five normal volunteers and four azotemic patients with secondary hyperparathyroidism who were not undergoing chronic hemodialysis. The normal persons and one azotemic patient took the drug for 5 weeks, and the remaining azotemic patients took it for 1 week. Before treatment, all patients had elevated levels of serum iPTH (two different assay systems), with or without elevated serum calcium concentrations, and increased urinary excretion of cAMP (per 100 ml glomerular filtrate). Cimetidine treatment caused no changes in serum calcium, phosphorus, or iPTH or in urinary cAMP (expressed as nanomoles per g creatinine). Serum creatinine, however, increased significantly in patients (P less than 0.02) and control subjects (P less than 0.025), which yielded statistically significant but spurious increases of urinary cAMP when expressed per 100 ml glomerular filtrate. We conclude that short term cimetidine administration has no effect on parathyroid function in normal persons or those with azotemic hyperparathyroidism. Because of its confusing effect on serum creatinine and a possible (albeit rare) adverse effect on renal function, the drug should be used with caution in azotemic patients not yet requiring chronic dialysis. PMID- 6281295 TI - Inhibition of follicle-stimulating hormone receptor binding by circulating immunoglobulins. AB - We have characterized a circulating inhibitor of FSH receptor binding found in two patients with hypergonadotropic amenorrhea and myasthenia gravis. The inhibitor behaves as an immunoglobulin according to several criteria, including precipitation by 30% ammonium sulfate, migration on DEAE-cellulose chromatography, specific binding to protein A-Sepharose, characterization as a 7S protein in sucrose density gradients, and immunoprecipitation with specific antihuman immunoglobulin G. Evidence suggests that these antibodies are directed at determinants on or near the FSH receptor, and they may be responsible for the observed clinical FSH resistance. PMID- 6281296 TI - Circulating levels of plasma adrenocorticotropin in polycystic ovary disease. AB - Excessive adrenal androgen production contributes to hyperandrogenism in polycystic ovarian disease (PCO). This study was performed to determine the concentration of basal plasma ACTH in PCO patients and normal women and correlate its level with that of circulating adrenal androgen. In PCO patients, significant increases in serum testosterone, androstenedione, and dehydroepiandrosterone sulfate were noted compared to levels in normal women. The mean circulating plasma ACTH in PCO patients (22 +/- 2 pg/ml) was not different from that in normal controls (20 +/- 2 pg/ml). The mean ratio of dehydroepiandrosterone sulfate to ACTH in individual PCO patients was significantly greater than that in normal subjects, whereas the cortisol to ACTH ratio was similar in both groups. These results suggest that increased adrenal androgen production in PCO patients is not due to abnormal ACTH secretion but arises from either altered adrenal responsiveness to ACTH or abnormal adrenal stimulation by a factor(s) other than ACTH. PMID- 6281297 TI - Effects of a met-enkephalin analog on adrenocorticotropin (ACTH), growth hormone, and prolactin in patients with ACTH hypersecretion. AB - The effect of D-Ala2, MePhe4, Met-(0)enkephalinol (Sandoz FK 33-824; 0.5 mg, im) on pituitary hormone secretion was studied in 11 patients with Addison's disease and 11 patients with ACTH-dependent Cushing's disease. In patients with Addison's disease, a pronounced fall of plasma ACTH levels was observed (P less than 0.005). The ACTH response to FK 33--824 was partially reversed by naloxone (4 mg, iv). In patients with Cushing's disease, no unequivocal decrease in either ACTH or cortisol was seen. Moreover, FK 33--824 failed to influence the vasopressin induced ACTH increase in 5 patients with Cushing's disease. In patients with cortisol deficiency due to either Addison's disease or bilateral adrenalectomy for Cushing's disease, FK 33--824 led to increases in PRL and GH similar to those described in normal subjects. However, in the presence of longstanding hypercortisolism, the PRL increase was significantly diminished, and the GH response to FK 33--824 was completely abolished. Our results suggest that in Addison's disease ACTH release is influenced by inhibitory opiate receptors. In patients with Cushing's disease, ACTH secretion is insensitive to FK 33-284, presumably because of an autonomous pituitary adenoma or hypothalamic derangement. The impairment of the PRL and GH responses to FK 33--824 in Cushing's syndrome seems to reflect a direct action of the elevated cortisol level, for it is not seen after bilateral adrenalectomy. PMID- 6281298 TI - Progesterone and adenosine 3',5'-monophosphate formation by isolated human corpora lutea of different ages: influences of human chorionic gonadotropin and prostaglandins. AB - From each of 24 women undergoing minilaparotomy at various stages of the luteal phase of the cycle, the corpus luteum (CL) was excised in toto, decapsulated, cut into pieces, and incubated for short time periods (5--120 min). Incubations were carried out in the absence and presence of hCG, prostaglandin F2 alpha (PGF2 alpha) and PGE2, both alone and in combination. Dating of the CL was done meticulously using several parameters. After incubation, the tissue levels of cAMP and the media concentrations of progesterone (P) were determined. The basal P production in vitro was highest in CL of the midluteal phase. hCG stimulated cAMP formation in CL of all ages, with the highest levels of cAMP being produced in CL of the midluteal phase. PGE2 was found to increase cAMP formation and potentiate the hCG effect in young CL, but not in CL of the midluteal phase. PGF2 alpha, alone or in combination with hCG, had no effect on cAMP or P formation in either young or old CL, while in CL of the midluteal phase, PGF2 alpha significantly counteracted the stimulatory effect of hCG on both cAMP and P formation. These in vitro data show that PGF2 alpha is capable of inducing functional luteolysis in humans. PMID- 6281299 TI - Deficient activity of receptor-cyclase coupling protein is transformed lymphoblasts of patients with pseudohypoparathyroidism, type I. AB - Erythrocytes of many patients with pseudohypoparathyroidism, type 1 (PHP-I) exhibit quantitatively reduced activity of the N protein, the guanine nucleotide binding regulatory component of adenylate cyclase. We have designated this group of patients PHP-Ia to distinguish them from PHP-Ib patients, in whom erythrocyte N activity is quantitatively normal. In virus-transformed lymphoblasts of three normal, three PHP-Ia, and two PHP-Ib subjects, we compared N and adenylate cyclase activities, as well as cAMP accumulation and susceptibility to radiolabeling in the presence of [32P]NAD and cholera toxin. In comparison to normal lymphoblasts, N activities were reduced by approximately 50% in lymphoblasts of the PHP-Ia patients, but were not reduced in lymphoblasts from the PHP-Ib patients. Toxin-catalyzed radiolabeling of the 42,000 molecular weight subunit of the N protein was also reduced in lymphoblasts of the PHP-Ia patients. These results are consistent with the hypothesis that N deficiency is generalized in tissues of PHP-Ia patients. Deficient lymphoblast N activity in PHP-Ia was not associated with decreases in adenylate cyclase activity or cAMP accumulation, probably because these activities involve many potential regulable cellular components in addition to the N protein. PMID- 6281300 TI - Homologous IM-9 lymphocyte radioreceptor and receptor modulation assays for human serum growth hormone. AB - Radioreceptor assays for human GH (hGH) have been developed using the IM-9 cultured human lymphoid cell receptor. Varying degrees of nonspecific interference with [125I] hGH binding to these cells occurs in the presence of serum. We have modified the traditional IM-9 competitive binding assay for hGH and abolished the nonspecific serum effects. The modified competitive assay is sensitive to as little as 2 ng/ml hGH, and it has been validated by the quantitative recovery of purified pituitary hGH in hypopituitary serum. Sera from stimulated normals, acromegalics, and patients with severe growth retardation were assayed. The RIA to radioreceptor assay ratios for these groups were 0.98 +/ 0.10, 0.97 +/- 0.18, and 2.43 +/- 0.54, respectively. The assay has potential usefulness in screening and predicting growth-retarded patients most likely to respond to exogenous hGH therapy. In addition, a sensitivity receptor modulation assay, which uses the ability of hGH to regulate its homologous IM-9 receptors, is described. This is 8- to 10-fold more sensitive than the nonregulatory assays and has been applied to the measurement of hGH in sera from unstimulated normals and acromegalics. The ratios of RIA to receptor modulation assay for the two groups were 1.17 +/-0.68 ad 1.07 +/- 0.26, respectively. These sensitive receptor assays offer a powerful technique for the measurement of biologically active and inactive GH peptide in serum, and may be particularly useful in the evaluation of statural growth disorders. PMID- 6281301 TI - Idiopathic hypercalciuria: hydrochlorothiazide decrease urinary calcium without altered renal response to parathyroid hormone. AB - Hydrochlorothiazide (HTZ) therapy reduces the urinary excretion of calcium (UCa V) in patients with idiopathic hypercalciuria (IH). To test the role of parathyroid hormone (PTH), namely the possibility that HTZ sensitizes the kidney to PTH as the sole or contributory cause of decreased UCa V associated with HTZ therapy, we measured urinary excretion of calcium (UCa V) and cAMP (UcAMP V), the tubular reabsorption of phosphate (TRP), plasma immunoreactive PTH, and the renal response to infused PTH (change in UcAMP V and TRP) in 10 patients with IH. Patients were studied before (control) and after 4 weeks of treatment with HTZ (100 mg/day). HTZ therapy significantly reduced UCa V (mean change, -122 +/- 19 mg/24 h). The UcAMP V, TRP, and plasma levels of calcium, phosphorus, and immunoreactive PTH were initially within the normal range and did not change significantly during HTZ therapy. PTH infusion resulted in a significant increase in the UcAMP V and a significant decrease in TRP during both control and HTZ therapy studies. There was, however, no significant difference in the degree of these renal responses to PTH infusion during the control vs. HTZ therapy studies. We conclude that the hypocalciuric effect of HTZ in patients with IH is independent of changes in PTH secretion, and that HTZ does not cause general sensitization of the nephron, namely proximal tubules, to PTH, as assessed by an increase in UcAMP V and a decrease in TRP. PMID- 6281302 TI - The use of theophylline as an in vivo probe of adrenocortical function. AB - The first step in the stimulatory action of most polypeptide hormones, including ACTH, is interaction with a specific target organ plasma membrane receptor. Theophylline, a nonspecific stimulus of several endocrine processes, does so presumably by circumventing the receptor step and directly increasing cAMP by inhibiting phosphodiesterase-mediated hydrolysis. Five patients with adrenal insufficiency, documented by a lack of cortisol secretion in response to exogenous ACTH, underwent a 4-h iv infusion of theophylline. In three of the five individuals, a significant concentration of cortisol was measured in serum for the first time. The patients who responded included one patient with the syndrome of ACTH insensitivity, one with ACTH deficiency, and one with idiopathic primary adrenal failure. Two patients with autoimmune adrenalitis failed to respond to theophylline, although one was tested very early in the course of her disease. We also noted that theophylline stimulated renin secretion and, in one patient with an intact zona glomerulosa, evoked a secondary rise in aldosterone equal to that produced by diuresis and upright posture. These studies suggest that the preservation of cortisol responsiveness to theophylline, after the loss of sensitivity to ACTH, may be relate to either the duration of the adrenal insufficiency or to the etiological mechanism. Patients with autoimmune adrenalitis may undergo more rapid and complete adrenocortical destruction, therapy losing sensitivity to both ACTH and theophylline, whereas patients with insufficient or ineffective ACTH stimulation may have receptor failure before the loss of intracellular function. Thus, responsiveness to iv theophylline may serve not only as a probe of potential adrenocortical reserve, but also as an indicator of pathogenesis. PMID- 6281303 TI - Receptor binding and processing of epidermal growth factor by human breast cancer cells. PMID- 6281304 TI - Glucocorticoid inhibition of cyclic AMP in the developing secondary palate. PMID- 6281305 TI - Detection of Japanese encephalitis virus immunoglobulin M antibodies in serum by antibody capture radioimmunoassay. AB - An assay for detecting human immunoglobulin M (IgM) antibodies to Japanese encephalitis (JE) virus was developed by using the antibody capture solid-phase radioimmunoassay approach (JE IgM ACRIAAA). Heavy-chain-specific goat antihuman IgM was first bound to the wells of a polyvinyl microtiter plate, and successive steps involved sequential binding of test sample IgM, acetone-extracted mouse brain JE antigen, and (125)I-labeled flavivirus hyperimmune human IgG. Among 20 patients hospitalized in Bangkok with clinical diagnoses of acute encephalitis, and with acute flavivirus infections proven by hemagglutination inhibition (HAI) serology, 16 had detectable (positive/negative [P/N] ratio, greater than 3.0) JE IgM ACRIA antibodies in the acute-phase serum specimen, and 19 had such antibodies in the convalescent-phase serum specimen. Convalescent patient sera regularly had higher P/N values than the corresponding acute-phase sera (mean +/- 1 standard deviation = 13.0 +/- 9.3 with acute-phase sera and 25.8 +/- 19.6 with convalescent-phase sera). JE virus-infected patients with HAI serological responses indicative of a primary flavivirus infection had higher JE IgM ACRI P/N responses than did those patients whose serological response indicated past exposure to other flaviviruses. None of 70 serum specimens from healthy Thai adults and children with serum JE HAI antibodies had detectable JE IgM ACRIA activity (P/N ratios all less than or equal to 3.0). Biological false-positives with low P/N ratios (range, 3 to 15) were found in sera from patients with acute or recent infections with flaviviruses other than JE virus but could be differentiated by the fact that these sera gave higher P/N ratios with homologous antigens than with JE virus. False-positive reactions with low P/N ratios (range, 3 to 6) due to serum rheumatoid factor activity were differentiated by testing with control antigen. The JE IgM ACRIA technique permits a rapid, accurate diagnosis of acute JE virus infections in both patients with and those without previous exposure to other flaviviruses. PMID- 6281306 TI - Rapid typing of herpes simplex virus based on immunological specificity of viral thymidine kinase and typing according to sensitivity to iododeoxyuridine. AB - We describe two methods for typing of herpes simplex virus (HSV). One procedure is based on the finding that the multiplication of HSV type 1 strains in primary rabbit kidney cells is inhibited by 2 x 10(-5) M iododeoxyuridine, whereas growth of HSV type 2 strains is considerably less affected. Forty-nine different HSV isolates were typed according to this method. For all isolates except two the results were found to be in agreement with results obtained by another typing procedure, the counterimmunoelectroosmophoretic method (S. Jeansson, Appl. Microbiol. 24:96-100, 1972). One HSV type 1 isolate behaved as a type 2 strain and was found to be a deoxythymidine kinase-negative mutant strain. The other deviant strain exhibited an intermediate iododeoxyuridine sensitivity, thus being impossible to type with this method. Another, faster typing procedure which is based on the immunological difference between HSV type 1 and 2 deoxythymidine kinase is also presented. This assay, in combination with the conventional methods for isolation, enables the detection of deoxythymidine kinase-negative therapy-resistant HSV strains. Finally, we report the detection and typing of HSV deoxythymidine kinase present in vesicle fluids. PMID- 6281307 TI - Solid-phase immune electron microscopy-double-antibody technique for rapid detection of papovaviruses. AB - The solid-phase immune electron microscopy-double-antibody technique, which takes less than 1 h to perform, was applied as a rapid, sensitive, and specific diagnostic tool in the demonstration of papovavirus particles. BK virus propagated in 82C human skin fibroblasts and a monospecific high-titer immune serum to BK virus were used to establish the test procedure. When Formvar-carbon coated grids were treated with appropriately diluted antibody, a 28-fold increase of virus particles per square micrometer was observed. Viewing of the virus particles was facilitated by the addition of a second "decorator" antibody. BK virus preparations at concentrations of 10(2) to 10(3) PFU/ml could be detected by this technique. There was no cross-reaction with mouse polyomavirus. PMID- 6281308 TI - Identification of parainfluenza virus serotypes by indirect solid-phase enzyme immunoassay. AB - An indirect enzyme immunoassay was used to detect and identify parainfluenza virus serotypes 1 and 3 in cell culture residuals from which infectious virus could no longer be recovered. PMID- 6281309 TI - Effect of parathyroid hormone on osmotic fragility of human erythrocytes. AB - The survival of erythrocytes (RBC) is shortened in uremia, and it has been shown that calcium influx into RBC evoked crenation and increased their rigidity. The high blood levels of parathyroid hormone (PTH) may augment entry of calcium into RBC and hence affect their integrity. We examined the effect of PTH on osmotic fragility of human RBC and investigated the mechanisms through which PTH interacts with RBC. Both the amino-terminal (1-34) PTH and the intact (1-84) PTH, but not the carboxy-terminal (53-84) PTH, produced significant increases in osmotic fragility. This effect was abolished by prior inactivation of the hormone. There was a dose-response relationship between both moieties of PTH and the increase in osmotic fragility. This action of PTH required calcium, was mimicked by calcium ionophore, and was partially blocked by verapamil. PTH caused significant influx of (45)Ca into RBC, which was not associated with potassium leak. The hormone did not affect water content of RBC. Scanning electron microscopy revealed that the incubation of RBC with PTH was associated with the appearance of membrane filamentous extensions, which anchor RBC together. Inhibition of glycolytic activity of RBC with NaF or inhibition of Na-K-activated ATPase with ouabain did not abolish the effect of PTH on osmotic fragility. PTH did not stimulate RBC Na-K-activated ATPase or Mg-dependent ATPase but caused marked and significant stimulation of Ca-activated ATPase. The basal activity of the RBC adenylate cyclase was low and PTH produced only a modest stimulation of this enzyme. Both cyclic AMP and dibutyryl cyclic AMP had no effect on osmotic fragility. THE DATA INDICATE THAT: (a) the RBC is a target organ for PTH, (b) the hormone increases osmotic fragility of RBC, and (c) this effect of PTH is due to enhanced calcium entry into RBC. We suggest that the increased calcium influx may affect the spectrin-actin of the cytoskeletal network of the RBC and may alter the stability and integrity of the cell membrane. This action of PTH on the RBC could be, at least in part, responsible for the shortened survival of RBC in uremia, and assign a new role for PTH in the pathogenesis of the anemia of uremia. PMID- 6281310 TI - Canine vascular tissues are targets for androgens, estrogens, progestins, and glucocorticoids. AB - Sex differences and steroid hormones are known to influence the vascular system as shown by the different incidence of atherosclerosis in men and premenopausal women, or by the increased risk of cardiovascular diseases in women taking birth control pills or men taking estrogens. However, the mechanisms for these effects in vascular tissues are not known. Since steroid actions in target tissues are mediated by receptors, we have looked for cytoplasmic steroid receptor proteins in vascular tissues of dogs. We find specific saturable receptors, sedimenting at 8S on sucrose density gradients for estrogens (measured with [3H]estradiol +/- unlabeled diethylstilbestrol), androgens (measured with [3H]R1881 +/- unlabeled R1881 and triamcinolone acetonide), and glucocorticoids (measured with [3H]dexamethasone +/- unlabeled dexamethasone); they are absent for progesterone (measured with [3H]R5020 +/- unlabeled R5020 and dihydrotestosterone). Progesterone receptors can, however, be induced by 1-wk treatment of dogs with physiological estradiol concentrations (100 pg/ml serum estrogen), indicating a functional estrogen receptor. Receptor levels range from 20 to 2,000 fmol/mg DNA. They are specific for each hormone; unrelated steroids fail to complete for binding. Low dissociation constants, measured by Scatchard analyses, show that binding is of high affinity. Steroid binding sites are in the media and/or adventitia since they persist when the intima is removed. Compared with the arteries, receptor levels are reduced 80% in inferior venae cavae of females, and are absent in the venae cavae of males. We hypothesize that steroid hormones can have direct effects on vascular tissues medicated by specific receptors present in arterial blood vessel walls. PMID- 6281312 TI - Effect of vitamin D status on the equilibrium between occupied and unoccupied 1,25-dihydroxyvitamin D intestinal receptors in the chick. AB - The dynamic equilibrium between in vivo occupied and unoccupied 1,25 dihydroxyvitamin D(3)[1,25(OH)(2)D(3)] receptors of the chick intestinal mucosa was investigated by the exchange assay previously reported [(1980). J. Biol. Chem.255: 9534-9537]. These parameters and their correlation to biological response, i.e., the levels of intestinal vitamin D-dependent calcium binding protein (CaBP), were assessed under different physiological conditions. After a single 1,25(OH)(2)D(3) injection (3.25 nmol), occupied receptor levels increased sharply to a maximum between 1 and 2 h, followed by a rapid decline. A single dose of 1alpha-hydroxy-vitamin D(3) [1alpha(OH)D(3)], an analog that requires 25 hydroxylation for biological activity, resulted in a protracted, albeit lower, response with maximal receptor occupancy at 6 h and half maximal levels 24 h after injection. The intestinal receptor occupancy patterns mirrored the serum 1,25(OH)(2)D(3) levels after either 1,25(OH)(2)D(3) or 1alpha(OH)D(3) treatment. Additionally, time-course (half-life) of blood disappearance of 1,25(OH)(2)D(3) and occupied receptor levels were similar (1.9 and 2.3 h, respectively), suggesting that the amount of occupied 1,25(OH)(2)D(3) receptor is determined by a simple equilibrium between serum 1,25(OH)(2)D(3) and unoccupied receptors. A dose-response study after intramuscular 1,25(OH)(2)D(3) injection yielded a hyperbolic curve with an apparent plateau at 70% receptor occupancy, corresponding to 5 nmol 1,25(OH)(2)D(3) injected. Half-maximal occupancy was reached after a dose of 1 nmol 1,25(OH)(2)D(3), corresponding to 1.5 ng 1,25(OH)(2)D(3)/ml serum. From this value the apparent K(d) in vivo is 3.7 nM, which is similar to that determined in vitro. A 10-fold increase in the 1alpha(OH)D(3) dose resulted in less than a doubling of the levels of serum 1,25(OH)(2)D(3), occupied 1,25(OH)(2)D(3) receptors, or CaBP. Under all experimental conditions, there was a positive correlation between occupied receptor and CaBP levels; however, the slope of the lines depended on the times chosen for the assays due in part to the lag period for CaBP induction and its accumulation within the cell. Conversely, the correlation between serum 1,25 (OH)(2)D(3) levels and occupied receptor levels yielded a single regression line independent of the observation time. Short and long-term treatment with different vitamin D metabolites, estrogen, progesterone, or cortisol did not affect the levels of total intestinal 1,25(OH)(2)D(3) receptor. Under normal physiological conditions, only 10-15% of the total 1,25(OH)(2)D(3) receptor population was occupied by ligand. These studies provide a basis for further investigations of physiological and biochemical parameters of the vitamin D endocrine system and their clinical applications. PMID- 6281311 TI - Increased numbers of alpha receptors in sympathetic denervation supersensitivity in man. AB - Cardiovascular responses to intravenous administration of norepinephrine and the properties of alpha receptors on platelets were compared in normal human subjects and subjects with multiple system atrophy (MSA) and sympathetic degeneration. All the subjects with MSA had low plasma norepinephrine concentrations (in the supine position) (0.42 +/- 0.09 nM, normal 3.47 +/- 0.58 nM), which did not increase on tilt. The pressor sensitivity of subjects with MSA to norepinephrine infusion was increased 10- to 20-fold, demonstrating denervation supersensitivity to adrenergic agonists. Analysis of alpha receptors was by binding of [3H]dihydroergocryptine to platelets. Results are shown as mean +/- standard error of the mean. In the MSA subjects, the number of alpha receptors (1,712 +/- 699 fmol/10(8) platelets) was about sevenfold greater than in normal subjects (224 +/- 21 fmol/10(8) platelets), and the affinity, as measured by the equilibrium dissociation constant (Kd), was similar in both groups (MSA subjects, 9.6 +/- 4.3 nM; normal subjects, 4 +/- 0.5 nM). These observations suggest that an increase in alphaadrenergic receptor numbers may account for the denervation supersensitivity to infused norepinephrine in patients with sympathetic degeneration. All the subjects with MSA had low levels of the endogenous adrenergic transmitter norepinephrine: the simultaneous increase in alpha adrenergic receptors supports the theory of agonist regulation of receptor numbers. PMID- 6281314 TI - Ultrasonic demonstration of intraductal biliary neoplasms. PMID- 6281313 TI - Neutropenia induced by systemic infusion of 5,12-dihydroxy-6,8,10,14 eicosatetraenoic acid: correlation with its in vitro effects upon neutrophils. AB - 5(S), 12(S)-Dihydroxy-cis-14,trans-6,8,10-eicosatetraenoate (compound I), 5(S),12(R)-dihydroxy-cis-14,trans-6,8,10-eicosatetraenoate (compound II), and 5(S),12(R)-dihydroxy-cis-6,14,trans-8,10-eicosatetraenoate (compound III) were prepared from rabbit peritoneal neutrophils challenged with arachidonic acid plus ionophore A23187. Each arachidonate metabolite caused rabbit neutrophils to aggregate and, in cells treated with cytochalasin B, release granule-bound enzymes. Compound III was 10- to 100-fold more potent than compounds II and I. When intravenously infused into rabbits at doses of 100--1,000 ng/kg, compound III induced abrupt, profound, transient neutropenia associated with a rapidly reversing accumulation of neutrophils in the pulmonary circulation. This in vivo action correlated closely with the ability of the fatty acid to activate neutrophils in vitro: neutropenia, aggregation, and degranulation occurred at similar doses of stimulus and the rapid, reversing kinetics of the neutropenic response paralleled the equally rapid, reversing formation of aggregates. The fatty acid did not alter the circulating levels of lymphocytes or platelets and did not aggregate platelets in vitro. At comparable doses (i.e., 100--1,000 ng/kg), compounds I and II did not cause neutropenia. Thus, compound III possesses a high degree of structural and target-cell specificity in stimulating neutrophils in vitro and in vivo. Clinical and experimental syndromes associating neutropenia with increased levels of circulating arachidonate metabolites may involve compound III as a mediator of neutrophil sequestration in lung. PMID- 6281315 TI - Oral ketoconazole for dermatophyte infections. AB - Nineteen patients with chronic dermatophyte infections were treated with 200 to 400 mg per day of ketoconazole orally. Sixteen had had their disease for 2 years or more. Sixty-three per cent achieved cure, and an additional 26% showed clinical improvement. Clearing of lesions occurred in 10 weeks (mean). The dose was increased to 600 mg per day in four patients. Of these, two achieved cure, for an overall cure rate of 74%. Seventy-five percent of patients who previously failed to respond to griseofulvin were cured or improved by ketoconazole. Four of eight patients with onychomycosis were cured, and the rest improved on therapy. One patient developed a generalized drug eruption after the first dose of ketoconazole, and possible minor side effects were noted in five. Three of eight patients followed off therapy after apparent cure subsequently showed relapse. The drug appears to have great potential value for the therapy of dermatomycoses. PMID- 6281316 TI - Technetium scanning in Kaposi's sarcoma and its simulators. AB - The clinical picture of ulcerated purple plaques on the legs often suggests several diagnoses: Kaposi's sarcoma, stasis dermatitis, atrophie blanche (livedoid vasculitis), and a poorly understood condition called acroangiodermatitis of Favre-Chaix (pseudo-Kaposi's sarcoma). Even the skin biopsy may not always be conclusive. We describe our experience with three patients with pseudo-Kaposi's sarcoma, one with "true" Kaposi's sarcoma and two with atrophie blanche. Clinical and histopathologic similarities among these three conditions pointed up the need for additional confirmatory studies, i.e., isotope scanning. The technetium scan was positive in both Kaposi's sarcoma and pseudo-Kaposi's sarcoma but negative in atrophie blanche. PMID- 6281317 TI - Ketoconazole: assessing its risks. PMID- 6281318 TI - Ketoconazole and the liver. PMID- 6281319 TI - Case report. Pseudomyxoma peritonei. PMID- 6281320 TI - Effect of nutrition on fertility and blood composition in the milk cow. AB - The relationship between nutrition, fertility and blood composition was studied in 400 cows from commercial farms. Fertility evaluated by overall conception rate (OCR) was shown to be significantly related proportion of concentrate and roughage feeds in the diet, and to its content of dry matter, digestible protein, Ca and P. The ratio of serum total lipids to glucose and a newly introduced parameter, delta Pi (a calculated value, obtained by subtracting whole blood levels of Pi from those of the blood serum), showed a highly significant relationship to OCR. No significant relationship was found between OCR and serum, total lipids, cholesterol, urea, Ca, Mg, organic and inorganic phosphate. The values and significance of the 2 blood parameters which showed a relationship to fertility are discussed. PMID- 6281321 TI - Addition of sodium bicarbonate to rations of postpartum dairy cows: physiological and metabolic effects. AB - Sodium bicarbonate was added to complete mixed rations to characterize physiological, metabolic, and ruminal changes immediately postpartum when dairy cows are switched abruptly from a low energy ration prepartum to a high energy ration postpartum. Twelve Holstein cows were paired and assigned randomly to either a control or buffered ration containing .8% sodium bicarbonate. Rations consisted of 50% corn silage:50% concentrate. All All cows were fed a similar dry cow ration for a minimum of 7 days prepartum and experimental rations for 2 wk beginning at parturition. Blood, feces, and urine were sampled on days 1, 2, 4, 7, 10, and 14 postpartum. Rumen fluid was sampled on days 7 and 14. Dry matter intake and milk production were 2.75% of body weight and 30.3 kg/day for cows fed buffer and 2.49% and 27.6 kg/day for cows fed control. Higher partial pressure of carbon dioxide and base excess in blood in cows fed buffer existed on days 2 and 4 postpartum than for cows fed the control ration. Cows fed buffer had higher concentrations of ruminal ammonia than cows fed control. This difference was less pronounced in blood urea nitrogen and urinary ammonia. Urine pH was higher for cows fed buffer than for control. Addition of sodium bicarbonate improved the acid-base status after abrupt change of ration and may be associated with increased dry matter intake and improved ration adaptation. Concentrations of most minerals and metabolites in blood serum did not differ between rations. PMID- 6281322 TI - Performance and health of offspring of cows experimentally exposed to polybrominated biphenyls. AB - Offspring of cows experimentally given polybrominated biphenyls(fire-Master BP-6) were studied from May 1976 through January 1981. Daily doses were 0,.25, and 250 mg for 60 days for Groups I, II, and III, and 250 mg for 180 or 202 days for Group V. Concentrations of polybrominated biphenyls in fat of dosed animals in Groups I, II, and III, and V were .05, .20, 25, and 30 micrograms/g at the end of dosing. Residue concentrations in body fat declined with a half life of about 1 yr. Seventy-five calves were born to these cows and their offspring, representing first through fifth parturition. Fifteen calves died, 10 of which were in Groups III and V. All deaths in Groups III and V were attributable to dystocia. Incidence od dystocia and calf mortality were related to birth weight of the calf which in turn was correlated with concentrations of polybrominated biphenyls in blood and tissues of the dam. Other variables that influenced birth weight were sex of calf, length of gestation, and parturition number of dam. There were no significant differences in growth and performance of calves from the four dosed groups. Clinical signs of polybrominated biphenyl toxicosis were not evident, and general health of offspring was not influenced. Number of inseminations for conception was greater among offspring from Groups III and V. Mechanisms for this dysfunction have not been determined. PMID- 6281323 TI - Oxytocin receptors and mammary myoepithelial cells. PMID- 6281324 TI - Clinical implications of drug-induced desensitization of the beta receptor after continuous oral use of terbutaline. PMID- 6281325 TI - Small cell lung carcinoma in the elderly. AB - Twenty-four patients over the age of 70 with small cell cancer of the lung were studied retrospectively. Ninety-two per cent of these patients had other concurrent medical disorders; 58 per cent had cardiac disease and 25 per cent had a second malignancy. The median survival of 20 treated patients was ten months, and the one-year survival was 30 per cent. Such elderly patients with multiple medical problems can benefit from combination chemotherapy. PMID- 6281326 TI - Linitis plastica involving the entire gastrointestinal tract. PMID- 6281327 TI - Focal dilatation of intrahepatic bile ducts (Caroli's disease), cholangiocarcinoma, and sclerosis of extrahepatic bile ducts: a case report. PMID- 6281328 TI - Linitis plastica: one disease or more? PMID- 6281329 TI - Plasmalemma-bound nucleoside diphosphatase as a cytochemical marker of central nervous system (CNS) mesodermal cells. AB - Nucleoside diphosphatase (NDPase) activity, using inosine-5'-diphospbate (IDP) as a substrate, was studied by light and electron microscope cytochemistry in brain and spinal cord of normal mouse. After cytochemical incubation for the enzyme, neurons, astroglial and oligodendroglial cells contained reaction product in the cisternae of endoplasmic reticulum (ER) and/or in the trans elements of the Golgi apparatus. In contrast, reaction product for NDPase was concentrated on the outer surface of the plasmalemma of microglial cells, endothelial cells of microvessels, pericytes, and smooth muscle cells of the arteriole wall. An occasional microglial cell contained sparse reaction product in the flattened saccules of the Golgi apparatus. The pattern of enzyme distribution for these cell types was the same for all parts of the nervous system examined in this study. These data suggest that in cells of ectodermal origin, NDPase is intracellular in location, whereas in cells of mesodermal origin, this enzyme is bound to the plasmalemma. The usefulness of this cytochemical reaction for NDPase as a marker of microglial and other cells in brain pathology is discussed. PMID- 6281330 TI - Dose-dependent effects of synthetic progestins on the biochemistry of the estrogenized post-menopausal endometrium. AB - Symptomatic post-menopausal women were treated with conjugated equine estrogens (Premarin; Ayerst International) 1.25 mg daily, continuously, adding either norethisterone (Primolut N; Schering Chemicals) 1, 2.5, 5 or 10 mg daily or D/L norgestrel (Wyeth Laboratories) 150 or 500 micrograms daily for 10 days each calendar month. Endometrial biopsies were obtained during estrogen therapy alone and on the 6th day of combined estrogen/progestin administration. Sensitive biochemical indices of estrogen and progestogenic activities were measured in th endometrial samples. These included measurements of DNA synthesis in epithelium and stroma by tritiated thymidine autoradiography; nuclear estradiol receptor content, and the activities of estradiol 17 beta and isocitric dehydrogenase and acid and alkaline phosphatase. Low dosages of progestins achieved maximal biochemical effects and the larger doses failed to enhance these responses. It is concluded that the dosages of progestins currently added in post-menopausal estrogen therapy are greatly in excess of those necessary to suppress endometrial proliferation effectively; lowering the daily progestin dosage is unlikely to result in any lessening of this protective effect and will probably reduce the incidence of side effects, as these appear to be dose-dependent. PMID- 6281331 TI - Induction of immunoglobulin secretion by a simple nucleoside derivative. AB - 8 bromoguanosine (8BrGuo), a low m.w. nucleoside derivative that rapidly traverses the lymphocyte membrane to the interior of the cell, is an extremely potent inducer of immunoglobulin production by B lymphocytes. It has recently been demonstrated that this same agent can initiate high level lymphocyte proliferation in vitro. Data in the current report demonstrate that 8Br-cGMP and other cGMP analogues are at best weak inducers of immunoglobulin secretion, being more than two orders of magnitude less effective (per molecule taken up) than 8BrGuo. Incubation of lymphocytes with 8BrGuo fails to elevate their intracellular concentrations of cGMP. Moreover, at equimolar concentrations, underivatized cGMP actually antagonized the induction of antibody production by 8BrGuo. These data, in concert with observations that many lymphocyte activators fail to alter cGMP content, that many agents that elevate cGMP fail to induce immunoglobulin synthesis, and that some cGMP-elevating agents even inhibit it, suggest that the induction of immunoglobulin production, like the induction of proliferation in B cells, is not primarily dependent upon cGMP. A simple nucleoside derivative is described, the use of which should prove to be a powerful probe for investigating the triggering mechanisms underlying the proliferative and differentiative B lymphocyte pathways at the molecular level. PMID- 6281332 TI - Spontaneous activation of the first component of human complement (C1) by an intramolecular autocatalytic mechanism. AB - For biochemical characterization, the first component of human complement (C1) was reconstituted from physiologic concentrations of purified C1q, 125I C1r, and 131I C1s. Upon incubation at 37 degrees C, C1 spontaneously activated, as evidenced by the characteristic proteolysis of the C1r and C1s polypeptide chains as detected by SDS-PAGE analysis. This spontaneous C1 activation followed first order kinetics (t 1/2 = 4 min and k = 0.173 min-1) with an activation energy of 19.1 kcal/mol. Spontaneous C1 activation was unaffected by the general protease inhibitor phenylmethylsulfonylfluoride (PMSF) but reversibly blocked by a known inhibitor of C1 activation, nitrophenylguanidinobenzoate (NPGB). Spontaneous C1 activation was measured at C1 concentrations ranging from 9 to 160 nM (i.e., 0.05 to 1.0 times physiologic concentrations). The data indicate that C1 spontaneously activates by an intramolecular autocatalytic mechanism, for first-order kinetics were observed over the entire concentration range with t 1/2 = 4 min at each concentration. However, the percentage of activable C1 decreased with dilution due to C1 dissociation (i.e., C1qr2s2 leads to C1q + C1r2s2). The observed concentration of C1 that spontaneously activated at each dilution equalled the concentration of C1 present as macromolecular C1. When reconstituted C1 was mixed with normal human serum (NHS) and then incubated at 37 degrees C, spontaneous C1 activation was completely inhibited. Pretreating NHS at 56 degrees C for 30 min destroyed its inhibitory activity. In conclusion, C1 spontaneously autoactivates at 37 degrees C by an intramolecular mechanism. This activation is suppressed in NHS. PMID- 6281333 TI - Stimulation of a human polymorphonuclear leukocyte oxidative response by the C1q subunit of the first complement component. PMID- 6281334 TI - Eosinophil peroxidase-mediated inactivation of leukotrienes B4, C4, and D4. AB - The slow-reacting substance (SRS) bioactivity of leukotrienes C4 (LTC4) and D4 (LTD4) was rapidly decreased by incubation with eosinophil peroxidase (EPO), H2O2, and iodide, bromide, or to a lesser degree, chloride, LTB4 chemotactic activity was also decreased by the EPO-H2-H2-halide system, although at a slower rate. Myeloperoxidase could substitute for EPO in these reactions. Leukotriene inactivation was greatly decreased or abolished by deletion of any of the components of the system or by the addition of the hemeprotein inhibitors, azide, cyanide, or aminotriazole, indicating a requirement for peroxidase. The H2O2 concentration employed in the above studies was 10(-4) M. H2O2 at higher concentrations (5 x 10(-4) to 10(-2) M) inactivated LTC4 and LTD4 in the absence of EPO and a halide but had no effect on the chemotactic activity of LTB4. We have previously shown that horse eosinophils stimulated with the calcium ionophore A23187 generate SRS. In the present study, eosinophils stimulated in this way were found to release extracellularly both H2O2 and EPO. Incubation of eosinophils with azide that inhibits EPO, and catalase that degrades H2O2, significantly increased the amount of SRS activity detected in the extracellular medium after A23187 stimulation. These findings suggests eosinophils may play an important modulating role in hypersensitivity reactions both by the production of leukotrienes and by their inactivation through the release of H2O2 and EPO. PMID- 6281335 TI - Delineation of four antigenic sites on a paramyxovirus glycoprotein via which monoclonal antibodies mediate distinct antiviral activities. AB - Six hybridoma antibodies specific for the hemagglutinin-neuraminidase (HN) molecule of the parainfluenza type 1 virus strain 6/94 were used to demonstrate the existence of four distinct antigenic sites on the HN molecule. Three of the sites (A, B-C, D) are topologically nonoverlapping, because antibodies to these sites bind noncompetitively to the HN molecule. Two sites (B, C) are operationally nonoverlapping, because mutations in site B do not detectably modify the antigenic site C. Although antibodies to each site had similar potencies (activity per microgram of antibody) in hemagglutination inhibition tests, antibodies to sites A and C or D differed approximately 100-fold in their potency to neutralize virus. Also, the antibody to site A strongly inhibited viral neuraminidase activity, whereas antibodies to sites C and D (and to a lesser extent to site B) enhanced the neuraminidase activity. Lastly, only antibodies to sites C and D formed precipitates in Ouchterlony double diffusion against detergent-disrupted virus. Because all six anti-HN antibodies are of IgG isotype and exhibited similar avidity for HN, these findings suggest that the ability of anti-HN antibodies to interact with the viral protein and to alter viral functions is largely dependent on their fine specificity. PMID- 6281336 TI - Isolation and characterization of circulating feline leukemia virus-immune complexes from plasma of persistently infected pet cats removed by ex vivo immunosorption. AB - IgG and circulating IgG immune complexes (CIC) were purified from plasma of three pet cats persistently infected with feline leukemia virus (FeLV) by adsorption to, and elution from, Staphylococcus aureus Cowan I. CIC were then separated from free IgG by sucrose gradient ultracentrifugation and were analyzed for the presence of FeLV structural proteins and corresponding specific antibodies. Radioimmunoprecipitation analysis indicated that FeLV envelope (gp70) and major core (p30) proteins, along with cat IgG heavy and light chains, were present in the CIC from all three cats. Further analysis of the CIC from one of the cats also revealed the presence of FeLV core proteins p15 and p12. IgG purified from isolated CIC was also shown to bind specifically to purified FeLV gp70, p30, and p15. These data provide direct evidence for FeLV-specific CIC in the plasma of persistently viremic pet cats, and suggest these animals are immunologically response to the virus even though free antibodies against the major structural proteins cannot be demonstrated in standard assays. PMID- 6281338 TI - Chondroid syringoma of skin. PMID- 6281337 TI - Relationship between the rearrangement of immunoglobulin genes, the appearance of a B lymphocyte antigen, and immunoglobulin synthesis in murine pre-B cell lines. AB - Eighteen Abelson virus-transformed immature B cell lines were established and immunoglobulin biosynthesis, expression of a B lymphocyte antigen detected by a monoclonal antibody, and rearrangement of immunoglobulin genes in these cell lines were studied. Only one cell line (A1) synthesized micro-chains but no light chains, and the other cell lines synthesized no detectable immunoglobulins. None of the cell lines established had detectable membrane-associated IgM. Fifteen cell lines expressed a B lymphocyte antigen on their cell surfaces. In three cell lines, however, the majority (greater than 99%) of cells did not express this antigen. Heavy chain genes were rearranged on both chromosomes in all the cell lines, although one heavy chain gene was deleted in three cell lines. In 12 of 18 cell lines, one or both kappa-chain genes were rearranged. In six cell lines, however, both kappa-chain genes remained in embryonic form; lambda-chain genes were in embryonic form in all the cell lines. These results suggested the hierarchy of Ig gene rearrangements, beginning with mu and proceeding to kappa and then to lambda. JH rearrangement was also shown to precede the appearance of a B lymphocyte antigen. In three cell lines (A1-A3), which were considered subclones derived from a single common precursor, it was suggested that one rearranged JH gene was functional, and the other was nonfunctional, indicating that allelic exclusion already operated in pre-B cells. PMID- 6281339 TI - Infection with cytomegalovirus during pregnancy: specific IgM antibodies as a marker of recent primary infection. AB - Specific IgM antibodies that persisted for up to four months were detected by radioimmunoassay (RIA) in the sera of 16 (55%) of 29 women with primary infections due to cytomegalovirus (CMV). The RIA for IgM detected primary infections in six (86%) of seven sera obtained within four months of seroconversion. In contrast, IgM antibodies were never detected by RIA in 104 serum samples from 18 women with recurrent infections due to CMV, irrespective of whether intrauterine transmission of virus had occurred. Specific IgM antibodies were also detected in the earliest samples during pregnancy of serum from three (14%) of 21 women whose type of infection with CMV was unknown but who had been delivered of congenitally infected infants. All of these results show that primary infection with CMV in the first trimester of pregnancy can be diagnosed by testing a single serum sample by RIA for IgM antibodies. Attempts to measure IgM antibodies by immunofluorescence gave a high frequency (19 [18%] of 104) of false-positive reactions. PMID- 6281340 TI - Activation of latent murine cytomegalovirus in vivo and in vitro: a pathogenetic role for acute infection. AB - Many cytomegalovirus (CMV) infections result from activation of virus previously latent in the host. Murine models of latent CMV infection have been developed in which latent virus can be activated in vivo by immunosuppression or by coculture of splenic lymphocytes in vitro. In the present study, latent murine CMV (MCMV) could be activated from lymphocytes of mice regardless of genetic strain, age at time of virus inoculation, or use of syngeneic or allogeneic fibroblasts for coculture. After intraperitoneal inoculation, virulent virus was activated from lymphocytes more often than attenuated MCMV (69% vs. 20% of lymphocyte cocultures were positive, respectively). Latent MCMV was not detected in lymphocytes after subcutaneous inoculation of weanling mice but could be activated from mice infected subcutaneously as newborns. The absence of latent infection in the lymphocytes of the mice that had been inoculated as weanlings was due to lack of virus replication in the spleen during acute infection. PMID- 6281341 TI - Effects of vaccination on cytomegalovirus-associated interstitial pneumonia in strain 2 guinea pigs. AB - The pathogenesis and prevention of generalized infection and interstitial pneumonia due to cytomegalovirus (CMV) in inbred strain 2 guinea pigs were investigated. Eighteen animals inoculated with approximately 10(7) 50% tissue culture infective doses of virulent salivary gland-passaged guinea pig CMV (GPCMV SG) developed generalized, viremic CMV infections, and 44% of the animals died. Most (68%) cocultivated tissues showed CMV infection, including lungs, which demonstrated severe bilateral interstitial pneumonia. Guinea pigs inoculated with live tissue culture-passaged GPCMV (GPCMV-TC) vaccine developed mild, nonlethal, generalized CMV infections and minimal histopathologic changes in infected tissues, including lungs. After challenge of 19 GPCMV-TC-vaccinated animals with GPCMV-SG, viremia was detectable in two (11%); CMV was detected in 15% of cocultivated tissues and rarely in lungs. Although vaccinated animals showed histologic evidence of interstitial pneumonia after challenge, none died. Strain 2 guinea pigs are useful for studies of pathogenesis and prevention of generalized CMV infection, particularly CMV-associated interstitial pneumonia. PMID- 6281342 TI - Pivmecillinam and relapse of typhoid fever. PMID- 6281343 TI - From the National Institutes of Allergy and Infectious Diseases. Summary of a workshop on antiviral agents for genital herpesvirus infections. PMID- 6281344 TI - [Substrate specificity of cyclic GMP-dependent protein kinase (author's transl)]. PMID- 6281345 TI - [Collagenase activities in healthy and inflamed gingivae induced by plaque in dogs. II. collagenase and gelatinase activities in dog gingiva after short term plaque accumulation (author's transl)]. PMID- 6281346 TI - [Observation on some clinical aspects of trophoblastic diseases (author's transl)]. PMID- 6281347 TI - [Studies on sintering of dental porcelain. (Part 2) Binary system sintering of quartz and low fusing frit (author's transl)]. AB - The effect of quartz addition was investigated on the sintering processes of powder compacts made from frit and glass powder with quartz powder. The results obtained are summarized as follows. 1) The quartz powder gave some effects on the shrinkage curves and the firing temperature range of frit and glass powder. Pyrex glass powder was more effective on the firing temperature range of the powder compacts with quartz addition comparing with other frits and glass powders. 2) The produced crystal during heat treatment of the powder compacts made from pyrex glass powder with quartz powder were cristobalite. 3) The sintered composite showed larger thermal expansion value by expansion due to the transition of cristobalite followed with the crystallization of cristobalite than that without crystallizing property. PMID- 6281349 TI - Embolization in otolaryngology. PMID- 6281348 TI - Relation of respiratory burst and arachidonate metabolism during phagocytosis by guinea pig alveolar macrophages. AB - We have examined the respiratory burst and arachidonic acid oxygenation that accompany phagocytosis in macrophages. Guinea pig alveolar macrophages were stimulated with opsonized zymosan in the presence of inhibitors of arachidonic acid metabolism: ASA, indomethacin, and ETYA, ASA, at concentrations as high as 60 micrograms/ml, had no effect on either oxygen consumption or superoxide ion formation. Indomethacin (4 x 10(-4) M) and ETYA (2 x 10(-5) M) did inhibit oxygen utilization and superoxide production. However, no indomethacin or ETYA inhibition of oxygen utilization was detected in the presence of 1 mM KCN, suggesting that the inhibitable portion of the respiratory burst observed with indomethacin or ETYA was dependent on mitochondrial respiration. Further study with ETYA showed that the inhibitor at 2 x 10(-5) M had little effect on uptake of 125I-labeled zymosan but did abolish the conversion of 14C-arachidonic acid to a compound that co-migrated with authentic 12-HETE on silica gel plates. Lower concentrations of ETYA (5 x 10(-6) M), which had no effect on the respiratory burst of phagocytosing alveolar macrophages, also inhibited arachidonic acid metabolism. We conclude therefore that the inhibition of oxygen consumption and superoxide production by ETYA at 2 x 10(-5) M is unrelated to inhibition of arachidonic acid metabolism. Furthermore, the oxygenation of arachidonic acid requires little of the oxygen consumed by phagocytosing alveolar macrophages. PMID- 6281350 TI - Dietary fiber. AB - Dietary fiber is plant-derived material that is resistant to digestion by human alimentary enzymes. Fiber may be divided into two broad chemical classes: 1) non alpha-glucan polysaccharides (cellulose, hemicelluloses, and pectins) and 2) lignins. Dietary fiber behaves within the gastrointestinal tract as a polymer matrix with variable physicochemical properties including susceptibility to bacterial fermentation, water-holding capacity, cation-exchange, and adsorptive functions. These properties determine physiological actions of fiber and are dependent on the physical and chemical composition of the fiber. Fiber undergoes compositional changes as a consequence of bacterial enzymatic action in the colon. Dietary fiber is of clinical significance in certain disorders of colonic function and in glucose and lipid metabolism. Dietary fiber increases stool bulk by acting as a vehicle for fecal water and by increasing fecal bacterial volume. Use of fiber in the treatment of constipation and uncomplicated diverticular disease is well established. By increasing stool bulk, fiber also reduces the fecal concentration of bile acids and other substances. Certain types of fiber decrease the rate of glucose absorption and attenuate postprandial rises in blood glucose and insulin. Plasma cholesterol levels are reduced by mucilaginous forms of fiber. This effect appears to be mediated in part by an increase in fecal acidic sterol excretion. PMID- 6281351 TI - Measurement of eicosapolyenoic acids in the serum by gas-liquid chromatography chemical ionization mass spectrometry. AB - The use of octadeutero eicosatetraenoic acid as an internal standard for the reproducible measurement of serum eicosapolyenoic acids (C20:3, C20:4, and C20:5) by gas-liquid chromatography-chemical ionization mass spectrometry is described. The method has the following advantages. The physicochemical properties of the internal standard and the eicosapolyenoic acids are similar. The acids are easily separated from compounds of similar retention times by means of selected ion monitoring. The measurements can be made more rapidly, (10 min per sample) than with previous techniques. PMID- 6281353 TI - Granular cell tumor of the buccal mucosa: a case report. PMID- 6281352 TI - The characterization of lipoproteins in the high density fraction obtained from patients with familial lecithin:cholesterol acyltransferase deficiency and their interaction with cultured human fibroblasts. AB - Lipoproteins of density 1.063--1.21 g/ml were isolated from the plasma of three sisters of Irish origin with familial LCAT deficiency. Fractionation of the lipoproteins on the basis of particle size by chromatography on Sephacryl S-300 permitted partial separation of two major and at least three other minor components which differed in their lipid:protein ratio and their apolipoprotein content. One of the major components was a small spherical lipoprotein whose sole apolipoprotein was apoA-I; the second major component contained predominantly apoA-I, together with apoE, and in addition, an apolipoprotein of molecular weight 46,000 that was not cleaved by reduction of disulfide bonds, and which was identified as apoA-IV. This apoprotein has not previously been detected in the lipoproteins of LCAT-deficient patients. A second apoE-containing lipoprotein, which contained apoA-I and apoE in a ratio of approximately 2:1, was also present as a minor component, together with two or more minor components whose apoproteins were comprised of apoA-I and apoC. The apoE-containing lipoproteins competed efficiently with 125I-labeled LDL for binding to high affinity LDL receptor sites on the surface of cultured human skin fibroblasts. The ability to bind to the LDL-receptor was directly proportional to the apoE content of the lipoproteins, even when other apoproteins, with the exception of apoB, were present in relatively large proportions. ApoE-containing 125I-labeled lipoproteins from an LCAT-deficient subject were also taken up and degraded by the cultured cells. PMID- 6281354 TI - Sealing ability of a new root canal filling material. PMID- 6281355 TI - The effects of tetraethylammonium and other agents on the potassium mechanoreceptor current in the ciliate Stylonychia. AB - The membrane response to mechanical stimulation was investigated under voltage clamp conditions in the hypotrich ciliate Stylonychia mytilus (Protozoa). The mechanoreceptor current shows high ionic selectivity, unlike than in metazoans. It is exclusively carried by potassium ions and is selectively inhibited by tetraethylammonium and 4-aminopyridine. Procaine, tetramethylammonium, caesium and divalent cations have no or little effect on this receptor current. The ionic channels mediating the currents differ from voltage-dependent channels in the ciliate membrane in their low sensitivity to divalent cations, and their relatively high sensitivity to tetraethylammonium and 4-aminopyridine. PMID- 6281356 TI - The presence of 17 beta-hydroxysteroid dehydrogenase activity in preimplantation rat and mouse blastocysts. AB - When Day 5 rat blastocysts and Day 4 and 5 mouse blastocysts were cultured in 53 microliters of medium containing 1340 or 2680 pg [3H]estradiol (E2), large amounts of [3H]estrone (E1) were detected in the medium at daily intervals for up to 5 days. This indicates the presence of 17 beta-hydroxysteroid dehydrogenase in the embryos. The activity was higher at a higher concentration of E2 and was also higher in mouse than in rat blastocysts. In the mouse, the activity was higher in Day 5 than Day 4 blastocysts during the first day in culture; then it decreased in Day 5 but increased in Day 4 blastocysts. The importance of E2 in embryonic development and implantation as suggested by others may be related to the activity of 17 beta-hydroxysteroid dehydrogenase. PMID- 6281357 TI - Effect of N-bromoacetamide on single sodium channel currents in excised membrane patches. AB - We have studied the effect of N-bromoacetamide (NBA) on the behavior of single sodium channel currents in excised patches of rat myotube membrane at 10 degree C. Inward sodium currents were activated by voltage steps from holding potentials of about -100 mV to test potentials of -40 mV. The cytoplasmic-face solution was isotonic CsF. Application of NBA or pronase to the cytoplasmic face of the membrane irreversibly removed sodium channel inactivation, as determined by averaged single-channel records. Teh lifetime of the open channel at -40 mV was increased about 10-fold by NBA treatment without affecting the amplitude of single-channel currents. A binomial analysis was used both before and after treatment to determine the number of channels within the excised patch. NBA was shown to have little effect on activation kinetics, as determined by an examination of both the rising phase of averaged currents and measurements f the delay between the start of the pulse and the first channel opening. Our data support a kinetic model of sodium channel activation in which the rate constant leading back from the open state to the last closed state is slower than expected from a strict Hodgkin-Huxley model. The data also suggest that the normal open channel lifetime is primarily determined by the inactivation process in the voltage range we have examined. PMID- 6281358 TI - Alamethicin channels incorporated into frog node of ranvier: calcium-induced inactivation and membrane surface charges. AB - Alamethicin, a peptide antibiotic, partitions into artificial lipid bilayer membranes and into frog myelinated nerve membranes, inducing a voltage-dependent conductance. Discrete changes in conductance representing single-channel events with multiple open states can be detected in either frog node or lipid bilayer membranes. In 120 mM salt solution, the average conductance of a single channel is approximately 600 pS. The channel lifetimes are roughly two times longer in the node membrane than in a phosphatidylethanolamine bilayer at the same membrane potential. With 2 or 20 mM external Ca and internal CsCl, the alamethicin-induced conductance of frog nodal membrane inactivates. Inactivation is abolished by internal EGTA, suggesting that internal accumulation of calcium ions is responsible for the inactivation, through binding of Ca to negative internal surface charges. As a probe for both external and internal surface charges, alamethicin indicates a surface potential difference of approximately -20 to -30 mV, with the inner surface more negative. This surface charge asymmetry is opposite to the surface potential distribution near sodium channels. PMID- 6281360 TI - Molecular homology and incompatibility in the IncFI plasmid Group. AB - The usual grounds for the inclusion of a plasmid in a particular incompatibility group are its mutual incompatibility with a type plasmid of that group, and, in some cases, the demonstration of shared regions of specific homology, presumed to be related to DNA replication. We have found that some plasmids classified as IncFI on genetical grounds share no homology with the previously described incompatibility regions of F on the basis of hybridization of specific radioactive probes to restriction enzyme digests of DNA from these plasmids. Others show homology with some or all of the regions of the F plasmid that can express incompatibility. The incompatibility behaviour of these plasmids has been examined to determine the relationship between the possession of regions of homology and the expression of incompatibility. Three plasmids, ColV3-K30, pHH507 and Entp307, show homology only with the secondary replicon of F and appear to use sequences homologous with the secondary F replicon in their replication. The results are consistent with the propositions that some contemporary IncFI plasmids arose by the integration of several replicons, and, in general, the replicon not being used for replicon expresses its incompatibility, as does the replicon being used for replication. We conclude that incompatibility of two plasmids with F does not necessarily demonstrate relatedness of the plasmids to each other, and that inclusion within the IncFI group can result from the possession of any of several combinations of inc sequences. PMID- 6281359 TI - Sodium and potassium fluxes and membrane potential of human neutrophils: evidence for an electrogenic sodium pump. AB - Sodium and potassium ion contents and fluxes of isolated resting human peripheral polymorphonuclear leukocytes were measured. In cells kept at 37 degrees C, [Na]i was 25 mM and [K]i was 120 mM; both ions were completely exchangeable with extracellular isotopes. One-way Na and K fluxes, measured with 22Na and 42K, were all approximately 0.9 meq/liter cell water . min. Ouabain had no effect on Na influx or K efflux, but inhibited 95 +/- 7% of Na efflux and 63% of K influx. Cells kept at 0 degree C gained sodium in exchange for potassium ([Na]i nearly tripled in 3 h); upon rewarming, ouabain-sensitive K influx into such cells was strongly enhanced. External K stimulated Na efflux (Km approximately 1.5 mM in 140-mM Na medium). The PNa/PK permeability ratio, estimated from ouabain insensitive fluxes, was 0.10. Valinomycin (1 microM) approximately doubled PK. Membrane potential (Vm) was estimated using the potentiometric indicator diS C3(5); calibration was based on the assumption of constant-field behavior. External K, but not Cl, affected Vm. Ouabain caused a depolarization whose magnitude dependent on [Na]i. Sodium-depleted cells became hyperpolarized when exposed to the neutral exchange carrier monensin; this hyperpolarization was abolished by ouabain. We conclude that the sodium pump of human peripheral neutrophils is electrogenic, and that the size of the pump-induced hyperpolarization is consistent with the membrane conductance (3.7-4.0 microseconds/cm2) computed from the individual K and Na conductances. PMID- 6281361 TI - Genetic instability of R plasmids in relation to the shift of drug resistance patterns in Salmonella johannesburg. AB - Observation of the resistance of Salmonella johannesburg to the six drugs ampicillin (A), streptomycin (S), tetracycline (T), chloramphenicol (C), kanamycin(K) and sulphadiazine (Su) was made over the 7 years from 1973 to 1979. Strains with ASTCKSu- and ASCKSu- resistance patterns predominated in the years 1973-1975 and 1976-1979, respectively. These resistances were found to be mediated by autotransferring plasmids belonging to the incompatibility group FIme. The ASTCKSu-resistance plasmids were unstable, giving rise to deletion variants at a much higher frequency than ASCKSu-resistance plasmids either of natural origin or derived in vitro from the ASTCKSu-resistance plasmids. Thus, the ASCKSu-resistance plasmid might be a deletion variant of the ASTCKSu resistance plasmid. This is supported by the extensive similarity of their cleavage patterns produced by specific restriction endonucleases. PMID- 6281362 TI - Envelope protein synthesis and inhibition of cell division in Escherichia coli during inactivation of the B subunit of DNA gyrase. AB - The rates of synthesis of inner and outer membrane proteins of Escherichia coli K12 during inhibition of cell division have been studied. When cell division was inhibited, either by treatment of wild-type cells with the antibiotic clorobiocin (an inhibitor of the B subunit of DNA gyrase) or by a temperature shift of a gyrB ts mutant, a 40% reduction in the rate of synthesis of total outer membrane protein relative to that of the inner membrane was observed. When a gyrB-ts mutant was shifted to high temperature under conditions which allowed continued cell division, this large reduction in the rate of synthesis of outer membrane protein relative to inner membrane protein was not observed. In contrast to the results obtained with clorobiocin, inhibition of cell division by the beta-lactam antibiotic cefuroxime did not cause any detectable disturbance in the rate of synthesis of either inner or outer membrane protein. This demonstrates that inhibition of septum formation per se does not perturb synthesis of envelope protein. The data obtained are consistent with a model in which the rate of synthesis and therefore expansion of outer membrane is one of many conditions which must be satisfied before septum formation can occur. The results are discussed in relation to such a model, and to previous findings which have shown that the rate of synthesis of outer membrane proteins displays a linear mode with an abrupt doubling in rate at a discrete point in the cell cycle. PMID- 6281363 TI - Identification and isolation of actin from Neurospora crassa. AB - Crude cell extracts of Neurospora crassa contained an abundant protein that was identified as actin by a number of criteria. The protein, either in cell extracts or in pure form, co-migrated with rabbit skeletal muscle actin in polyacrylamide gels. The N. crassa actin was purified by DEAE-cellulose and DNAase I-Sepharose chromatography and had the expected property of inhibiting DNAase I activity. Although N. crassa actin could polymerize and depolymerize, purification based entirely on this characteristic was ineffective. The actin was susceptible to proteolytic degradation, and under certain conditions, a breakdown product of defined size was observed. PMID- 6281364 TI - Restriction enzyme analysis of mitochondrial DNA of members of the genus Aspergillus as an aid in taxonomy. AB - Restriction endonuclease patterns of mitochondrial DNA from seven species of the genus Aspergillus (A. nidulans, A. wentii, A. awamori, A. niger, A. oryzae, A. tamarii and A. echinulatus) have been compared in order to test the phylogenetic relationships between these species. The fraction of restriction fragments common between all pairs of species from this set ranged from 0 to 1. The use of this approach for the taxonomy of the genus Aspergillus is discussed. The postulated phylogenetic relationships differ significantly in two cases from the classification of the genus presented by Raper & Fennell (1965). PMID- 6281365 TI - Control of synthesis of wall teichoic acid during balanced growth of Bacillus subtilis W23. AB - Enzymes involved in the synthesis of teichoic acid and its linkage to the wall in Bacillus subtilis W23 were measured in chemostat cultures growing at equilibrium at a dilution rate of 0.2 h-1 in different concentrations of inorganic phosphate. All the enzymes, except teichoic acid glucosyl transferase, which was insensitive to changes in phosphate concentration, were almost undetectable at 0.5 mM phosphate. At higher phosphate concentrations the changes in activity of the enzymes of linkage unit synthesis were sufficient to account for the changes in the rate of incorporation of teichoic acid into the wall in vivo. Between 3.5 and 4.5 mM-phosphate the amount of teichoic acid synthesized in vivo increased, but no increase in the ability of toluenized bacteria to synthesize teichoic acid could be detected. Allosteric regulation might therefore be important at high phosphate concentrations. Bacteria maintained a constant ATP content and a constant adenylate energy charge during chemostat growth at all phosphate concentrations. PMID- 6281366 TI - A specialized transducing phage, lambda psrlA, for the sorbitol phosphotransferase of Escherichia coli K12. AB - A specialized transducing phage for the srlA gene, specifying the sorbitol specific Enzyme II of the phosphoenolpyruvate:sugar phosphotransferase system, was constructed and its DNA was analysed by restriction endonuclease digestion. Phage construction involved four steps: (1) integration of lambda into the srlA gene; (2) selection of phage carrying (a) the left and (b) the right end of the srlA gene by means independent of the function of the new DNA acquired; (3) reconstitution of the srlA gene in a dilysogen of these two phage; and (4) the excision, using the heteroimmune lambdoid phage 21, of a plaque-forming srlA+ phage from the dilysogenic chromosome. Comparison of the DNA restriction digests of the transducing phage with those of its parents and of wild-type lambda revealed fragments consisting partly of lambda and partly of Escherichia coli DNA. The junction points in the intermediate phage define a site that must lie within the reconstituted gene of the final phage. This technique should be of general application in relating genes, cloned by our method, to DNA sequences. PMID- 6281367 TI - Control mechanisms governing the infectivity of Chlamydia trachomatis for hela cells: modulation by cyclic nucleotides, prostaglandins and calcium. AB - Chlamydia trachomatis causes common infections of the eyes and genital tract in man. The mechanism by which this obligate intracellular bacterium is taken into epithelial cells is unclear. The results described here support the concept that chlamydial infections of HeLa cells is under bidirectional cyclic nucleotide control, with guanosine 3':5'-cyclic monophosphate (cGMP) acting as a stimulator, and adenosine 3':5'-cyclic monophosphate (cAMP) as an inhibitor. Treatment of the HeLa cells with the divalent cation ionophore A23187, with carbamoylcholine, or with prostaglandins known to increase the concentration of endogenous cGMP, also increased host cell susceptibility to chlamydial infection. Cyclic GMP was only effective if added at or before chlamydial inoculation, suggesting that its main effect was on chlamydial uptake. The stimulatory effect of cGMP, but nt antagonism, by cAMP, was abolished if the cells were first treated with any of four different inhibitors of prostaglandin synthesis, suggesting a critical role for endogenous prostaglandin synthesis. Centrifugation of chlamydiae on to host cells was followed by a rapid increase in the mobility of Ca2+ across the cell membrane. The interrelationships of these observations and the possibility that chlamydiae and other intracellular pathogens might evoke alterations in host cell prostaglandin and cyclic nucleotide concentrations to aid their own uptake are discussed. PMID- 6281368 TI - Aspects of thymidine metabolism and function in cultured mammalian cells infected with herpes simplex virus type 1. PMID- 6281369 TI - Fine structure of virus chromatin in thin sections of SV40-infected cells: a cytochemical and autoradiographical study. AB - The ultrastructural organization of virus chromatin was studied within nuclei prepared from CV1 cells (cultured monkey kidney cells) lytically infected with SV40 virus (simian virus 40) by a procedure which allows a mild loosening of nucleoproteins. In addition to dispersed host components, DNA-containing nucleoplasmic structures could be identified as virus chromatin. Both standard staining for structure and specific staining for DNA clearly revealed in thin sections the nucleosomal structure of well-extended virus chromatin as well as alignments of virions on host chromatin. In addition, replicating and transcribing virus chromatins were abundant as revealed by high-resolution autoradiography. Therefore, the procedure of loosening of nucleoproteins used in this report also preserves the active SV40 chromatin and allows the in situ visualization in Epon sections of transcription and replication of the virus genome. PMID- 6281370 TI - Three strains of European foot-and-mouth disease virus are highly conserved in the 3'-termini and highly variable in the genes of two capsid proteins. AB - Restriction enzyme-generated subgenomic fragments of cloned cDNA prepared from RNA of the strain O1 Kaufbeuren (O1K) of foot-and-mouth disease virus (FMDV) were compared qualitatively and quantitatively for sequence complementarity with radioactive RNA from strains C Oberbayern (CObb) and A2 Spain (A2S) in hybridization experiments on nitrocellulose membranes. Quantitative comparison of nucleic acid sequences neighbouring (C Obb/O1K) or including (A2S/O1K) the 3' end of the virus genomes demonstrated more than 80% homology. In contrast, sequences coding for the capsid proteins VP1 (10%, CObb/O1K; 16 to 21%, A2S/O1K) and VP3 (12%, A2S/O1K) were remarkably heterologous. Sequences downstream from the gene for VP1, i.e. those coding for non-structural proteins, showed 23 to 51%f homology to both RNAs except for the area coding for protein P56a. Here, the observed homology was 82% to CObb and 39 to 46% to A2S. PMID- 6281371 TI - Characterization of strains of type 3 poliovirus by oligonucleotide mapping. AB - T1 RNase oligonucleotide maps of RNA prepared from type 3 poliovirus were shown to be highly characteristic of the strain from which they were prepared. Strains isolated from paralytic cases of poliomyelitis temporally associated with live poliovirus vaccination were shown to be very closely related to the Sabin vaccine strain. Comparison of strains isolated in 1962, and strains isolated between 1973 and 1975 indicated that strains derived from the Sabin vaccine strain have displaced other circulating strains of type 3 poliovirus in the U.K. as a result of the use of live polio vaccine. PMID- 6281372 TI - Structural study of vesicular stomatitis virus G protein in the virion envelope. AB - Some structural properties of the vesicular stomatitis virus (VSV) G protein were examined in virions and isolated envelope fragments. We have shown that in the virion a portion of the G protein extends through the lipid envelope and that this part of the molecule can be cleaved by chymotrypsin. Envelope fragments isolated from VSV without the use fo detergents maintained the structural characteristics of the G protein found in intact virions. In addition, we provide evidence that at least some of the isolated envelope fragments have both sides of the bilayer exposed to added reagents, suggesting that this preparation would be useful for in vitro reassociation experiments. PMID- 6281373 TI - Slow virus replication: the role of macrophages in the persistence and expression of visna viruses of sheep and goats. AB - Lentiviruses of sheep and goats cause slowly progressive diseases of the central nervous system (visna), lungs (maedi) and joints (arthritis) in their natural hosts. However, the virus target cell(s) in these diseases are still unknown. In this report, using laboratory-adapted Icelandic visna virus and several field strains recently obtained from sheep and goats with natural disease in the U.S.A., we show that macrophages became persistently infected when inoculated in culture. Furthermore, macrophages were an invariable source of virus from experimentally and naturally infected animals. Virus-producing macrophages developed minimal cytopathic changes and virus assembly occurred mainly intracellularly, accumulating in cytoplasmic vacuoles. In contrast to macrophages, sheep choroid plexus fibroblasts developed syncytial cytopathic changes after inoculation and virus maturation occurred at the cell surfaces. Replication of the Icelandic virus was highly productive in this system but that of the field viruses was very inefficient. In some cases these agents failed to replicate in the fibroblasts and no cytopathic effect occurred. This block in the field virus replication was, however, overcome when infected nonproducer fibroblasts were co-cultivated with macrophages. In these cases, virus production with attendant cytopathic effect in the fibroblasts required the continuous presence of macrophages because the cells reverted to a non-productive state when separated from macrophages and became productive again when subcultures were added to new macrophages. The roles of the macrophage as a virus target cell and virus inducer in the virus-macrophage-fibroblast interactions are discussed with inferences to the well-known phenomenon of restricted virus replication in infected animals and the immunopathological aspects of the diseases. PMID- 6281374 TI - Specific cross-linking of capsid proteins to virus RNA by ultraviolet irradiation of poliovirus. PMID- 6281375 TI - Suppression of fibronectin synthesis by an early function(s) of human cytomegalovirus. AB - Labelling of fibronectin with [3H]leucine and its isolation by immunoprecipitation followed by electrophoresis on gels showed that fibronectin synthesis was specifically suppressed in human embryonic cells infected with human cytomegalovirus. Degradation or release of pre-existing fibronectin into the medium was not affected. The inhibition of fibronectin synthesis was also observed when cells were infected with a DNA-minus temperature-sensitive mutant at the non-permissive temperature but not observed in infection with u.v. irradiated virus, suggesting the involvement of the expression of an early virus function(s). This capacity of the virus may be implicated in the virus-induced early cell rounding. PMID- 6281376 TI - In vitro stimulation of rabbit T lymphocytes by cells expressing herpes simplex antigens. AB - Lymphocyte stimulation responses to herpes antigens were studied using virus infected X-irradiated cells. Rabbits were immunized with herpes simplex virus type 1 (strain HFEM) grown in RK 13 cells. For in vitro stimulation assay BHK21 cells were X-irradiated (15 000 rad) and infected with a high m.o.i. of a temperature-sensitive (ts) mutant (N102) of HFEM strain at the non-permissive temperature (38.5 degrees C) of virus. Virus antigens were expressed on the infected cells and there was no leakage of infectious virus into the medium at 38.5 degrees C. T lymphocytes from rabbits immunized with herpes simplex virus were specifically activated by herpesvirus-infected X-irradiated cells; lymph node cells from rabbits immunized with RK13 cells and from non-immune rabbits showed no proliferative response. PMID- 6281377 TI - Properties of monoclonal antibodies directed against the glycoproteins of Sindbis virus. AB - Four monoclonal antibodies that react with Sindbis virus glycoproteins have been examined for (i) their effects on virus infectivity, (ii) their ability to recognize conformational changes in glycoprotein structure, and (iii) their cross reaction with several different alphaviruses. Two of the monoclonal antibodies reacted with the native forms of the E1 glycoprotein but did not neutralize virus infectivity. One of the anti-E1 antibodies formed an infectious virus-antibody complex. The other two monoclonal antibodies reacted with the E2 glycoprotein in both an unfolded as well as a native structure. One of these antibodies reacted with Semliki Forest virus. The anti-E1, but not the anti-E2, antibodies cross reacted with three serologically related equine encephalomyelitis viruses. PMID- 6281378 TI - The differentiation of calf rotaviruses by plaque morphology and serum neutralization. PMID- 6281379 TI - Noradrenaline and dopamine-beta-hydroxylase levels in rat salivary glands after preganglionic nerve stimulation: evidence for re-use of amine storage granules in transmitter release. AB - The levels of endogenous noradrenaline (NA) in rat salivary glands were reduced to about 40 per cent after 1 hour of preganglionic stimulation at 4 Hz. Following 2 to 3 hours of rest the NA values had recovered to normal and remained normal for 6 hours after the stimulation. The recovered NA could be depleted by renewed nerve stimulation. After treatment with reserpine the NA disappeared almost completely. The recovered NA is discussed to be mainly present in amine storage granules that are re-used for storage and transmitter release. The axonal down transport of new granules to the nerve terminals appears to be too small, as based on earlier results, for being of any quantitative importance in the experimental situation. The dopamine-beta-hydroxylase (DBH) level in the glands did not significantly change after the period of stimulation. The DBH content remained mainly unchanged following either post-stimulation recovery, repeated stimulation, or reserpine treatment. First when the glands were stimulated for 4 hours, there was a small decrease of the DBH levels. The mainly unchanged levels of DBH after stimulation does not appear to be explained by "trapping" in the tissue of released DBH, or recovery of the DBH by a marked axonal down-transport of new granules, as based on earlier results. The results of the DBH estimations support the view that the granules are not consumed to any marked extent at nerve activity, but can be re-used for nerve transmission. Furthermore, the results are in agreement with lack of correlation between released NA and released DBH. PMID- 6281380 TI - Increasing external K+ blocks phase shifts in a circadian rhythm produced by serotonin or 8-benzylthio-cAMP. AB - Serotonin (5-HT) phase shifts the circadian rhythm from the isolated eye of Aplysia. The discovery of the mechanisms involved in phase shifting by 5-HT may help elucidate the nature of the circadian oscillator. We have found that 5-HT appears to phase shift by causing a change in membrane K+ conductance. Solutions containing zero K+(0-K+) phase shift the rhythm and the phase response curve (PRC) for 0-K+ is similar to one previously obtained for 5-HT. The similarity in PRCs for 0-K+ and 5-HT suggested that these treatments may be phase shifting the rhythm through a common mechanism. The nonadditivity of phase shifting by 0-K+ and 5-HT supports this suggestion. A common mechanism of action of 5-HT and 0-K+ might be effects on membrane potentials. The possible involvement of a membrane potential change in mediating the effect of 5-HT and the lack of an effect of large reductions in Na+, Cl-, and Ca2+ ions on phase shifting by 5-HT led us to examine the role of K+ ions in phase shifting by 5-HT. A change in K+ conductance may mediate the effects of 5-HT on the rhythm because HiK (30mM) solutions blocked the phase shift normally produced by 5-HT. The conductance change produced by 5-HT may be an increase in K+ conductance which would produce a hyperpolarization and not a decrease in K+ conductance which would produce a depolarization since depolarizing treatments, HiK (30-110mM), had no effect on the rhythm at the phase where 5-HT produces its largest phase shifts. Since we previously found that the effects of 5-HT appear to be mediated by cAMP, we examined whether HiK solutions could block the effects of 8-benzylthio-cAMP on the rhythm. HiK (40mM) blocked the phase shifts normally produced by 8-benylthio cAMP. Our working hypothesis for the 5-HT phase-shifting pathway based on these results is 5-HT leads to increased cAMP leads to elevates K+ conductance leads to membrane hyperpolarization leads to phase shifts the rhythm. PMID- 6281381 TI - Evidence for peptide-mediated neurotransmission in a molluskan brain. AB - The previous report (Snow, 1982) characterized the monosynaptic actions of an identified cerebral interneuron (C2) in the marine mollusk Tritonia. The C2 neurons produce four types of postsynaptic potentials in an identified pedal neuron (Pd5). A high-molecular-weight (approximately 1400 daltons by Sephadex G 15 gel filtration) compound, which could mimic the four postsynaptic responses in Pd5, was isolated from C2 somata. The C2 somata had the ultrastructural characteristics of peptide-secreting cells, including profuse rough endoplasmic reticulum and large (170 nm average diameter) dense secretory vesicles. These data are consistent with the hypothesis that the synaptic transmitter of C2 neurons is a peptide(s). PMID- 6281382 TI - Binding of 2-formylpyridine monothiosemicarbazonato copper II to cat and normal human hemoglobins. AB - The antitumor agent 2-formylpyridine monothiosemicarbazonato copper(II) forms adducts with sulfur and nitrogen donor atoms from cat hemoglobin but only nitrogen donor atoms from human hemoglobin. Improved resolution of the mI = 1/2 lines in the g parallel region at S-band not only confirms the number of nitrogen donor atoms in the square planar configuration but provides evidence for strong coupling from a proton. Adduct formation results in an increase in the oxygen affinity of hemoglobin. Thus, it is suggested that allosteric enzyme inhibition may be a mechanism for the action of this agent. PMID- 6281383 TI - Different behavior of sulfonamides with respect to copper-substituted bovine and human carbonic anhydrases. PMID- 6281384 TI - Opiate receptors from different tissue sources: solubilization and characterization. PMID- 6281385 TI - The effects of neonatal pedunclectomy on [3H]noradrenaline uptake and the development of beta-adrenergic receptors in the rat cerebellum. AB - A newly developed method for cutting the cerebellar peduncles in neonatal rats has allowed the study of the development of cerebellar beta-adrenergic receptors in the absence of noradrenergic afferents. Cutting the cerebellar peduncles of neonatal animals did not affect the pattern of development of the beta-adrenergic receptors, nor their final numbers. Pedunclectomy induced a decline in the ability of slices of cerebellar cortex to accumulate [3H]noradrenaline although high-affinity noradrenaline uptake, was never completely abolished. It is suggested that the remaining high-affinity noradrenaline uptake cannot be attributed to noradrenergic fibres from the locus coeruleus. PMID- 6281386 TI - Species differences in the brain regional distribution of receptor binding for thyrotropin-releasing hormone. AB - A survey of the regional distribution of binding of 1 nM [3H](3-Me His2)thyrotropin-releasing hormone ([3H]MeTRH) to TRH receptors in the brains of eight mammalian species revealed major species differences in both the absolute and relative values of TRH receptor binding in different brain regions. Several brain regions exhibited binding equal to or exceeding that in the anterior pituitary gland of the same species, including the amygdala in the guinea pig and rat, the hypothalamus in the guinea pig, the nucleus accumbens in the rabbit, and all these and other regions in the cat and dog, for which pituitary binding was exceptionally low. Species could be divided into two groups according to which brain region appeared highest in binding: rabbits, sheep, and cattle had highest binding in the nucleus accumbens/septal area, whereas guinea pigs, rats, dogs, cats, and pigs had highest binding in the amygdala/temporal cortex area. The nucleus accumbens consistently exceeded the caudate-putamen in receptor binding. For most brain regions, rabbits, rodents, and sheep tended to be higher than carnivores, cattle, or pigs. Further regions that exhibited appreciable binding in most species included the olfactory bulb and tubercle, hippocampus, and various cortical and brain stem areas. In fact, essentially all brain regions appeared to have detectable levels of TRH receptors in at least some species, but no rat peripheral tissues have yet shown detectable receptor binding. The species differences appeared to reflect largely if not entirely differences in receptor density, although this was not tested in every species. PMID- 6281387 TI - Acid sphingomyelinase of human brain: purification to homogeneity. AB - Acid sphingomyelinase (sphingomyelin phosphodiesterase, EC 3.1.4.12) was purified from human brain by extraction with 0.1% Triton X-100, followed by sequential chromatography on Concanavalin A-Sepharose, octyl-Sepharose, hydroxylapatite, DEAE-cellulose, red A-agarose, Sephadex G-200, and DEAE-cellulose with ampholyte elution. Sphingomyelinase activity was purified more than 20,000-fold from the starting homogenate with a 1% yield. Specific activity of up to 800 mumol/h/mg protein could be achieved. Gel electrophoresis with 6% polyacrylamide containing sodium dodecyl sulfate gave a single protein band with a molecular weight of 70,000, in good agreement with the molecular weight previously estimated from sucrose density gradient centrifugation in 0.1% Triton X-100. Triton X-100 could be readily removed from the enzyme by sucrose density gradient centrifugation. The Triton-free enzyme showed the same Km and pH optimum. Heat stability of the enzyme was reversibly affected by Triton X-100, in that removal of the detergent made the enzyme more heat labile. The Km of purified enzyme for sphingomyelin was 36 microM. It was unaffected by sulfhydryl reagents, but was inhibited by dithiothreitol at high concentrations. The preparation was free of all lysosomal hydrolase activities tested, including galactosylceramidase and alpha mannosidase, which tended to copurify in our previous procedure. The enzyme was inactive toward sphingosylphosphorylcholine. It was active with bis[p nitrophenyl]- and bis[4-methylumbelliferyl]phosphate and the chromogenic and fluorogenic sphingomyelin analogues. PMID- 6281388 TI - Inactivation of benzodiazepine binding sites by N-ethylmaleimide. AB - The benzodiazepine receptors of bovine brain membranes have been identified by the specific binding of radiolabeled [3H]diazepam. Pretreatment of membranes with N-ethylmaleimide causes a dose- and time-dependent decrease of 45 to 60% in the number of binding sites. No decrease occurs when membranes are pretreated with N ethylmaleimide before administration, or in the presence, of diazepam. Binding of [3H]diazepam to the remaining sites occurs with the same characteristics as binding to the untreated receptor population. PMID- 6281389 TI - Buprenorphine: high-affinity binding to dorsal spinal cord. AB - The binding of the mixed opiate agonist-antagonist [3H]buprenorphine was compared with [3H]naloxone and [3H]dihydromorphine binding in membranes prepared from rat whole brain and dorsal spinal cord. Scatchard analysis of binding to whole brain yielded KD values close to 1.0 nM for all three 3H-ligands studied, although [3H]buprenorphine labelled five times as many binding sites. [3H]Naloxone and [3H]dihydromorphine bound to dorsal spinal cord with approximately the same affinity as to whole brain, although both 3H-ligands labelled fewer sites in the spinal cord. In contrast, Scatchard analysis of [3H]buprenorphine binding to spinal cord yielded curvilinear Scatchard plots, suggesting the presence of a very high-affinity (KD = 0.12 nM) binding site in addition to the high-affinity site (KD = 1.0 nM) present in the brain. Studies on the displacement of [3H]buprenorphine by opiates and D-Ala2,Met5-enkephalinamide supported the presence of two binding sites for this ligand in the spinal cord. PMID- 6281390 TI - Effect of a synthetic pyrethroid deltamethrin on excitability changes following a nerve impulse. AB - Excitability changes following a nerve impulse were studied in the rat tail. Supernormal nerve excitability in control animals was present, as in other vertebrate nerve fibres, from 4-30 ms and was followed by a period of subnormal excitability extending up to 100-200 ms. Two to seven hours after intravenous administration of 1.5 mg/kg deltamethrin, supernormality was increased in degree and prolonged in duration for up to 400 ms. A minor effect was still detectable 24 hours after injection. This effect of deltamethrin on nerve excitability is probably due to persistent slight depolarisation of the nerve membrane resulting from its known effect of maintaining a proportion of sodium channels open for up to several hundred milliseconds following a nerve impulse. PMID- 6281391 TI - Familial extrapyramidal disease with peripheral neuropathy. AB - A family is presented in which 10 members over three generations developed a Parkinsonian syndrome. Two siblings had additional clinical and electrophysiological evidence of an associated peripheral neuropathy and sural nerve biopsy in one case revealed axonal loss. This is the third family reported in the literature in which an extrapyramidal syndrome is associated with neuropathy, but differs from earlier reports in that neuropathic involvement was due to axonal degeneration rather than segmental demyelination. PMID- 6281392 TI - The relationship between anti-acetylcholine receptor antibody levels and neuromuscular function in chronically myasthenic rats. AB - Rats immunized with acetylcholine receptor protein (AChR) purified from Torpedo electric organ develop defects of neuromuscular function closely mimicking those of myasthenia gravis. These rats are easily fatigued, have reduced neuromuscular transmission, and high titers of anti-AChR antibody. Contrary to previous findings however, the present study finds a very good correlation between the development of the anti-rat AChR antibody levels and subsequent decreases in neuromuscular function in experimentally myasthenic animals. This result supports the hypothesis that anti-AChR antibody levels play a central role in development of experimental myasthenia. The relevance of these findings to the pathology of myasthenia gravis is considered. PMID- 6281393 TI - Neuron to neuron transmission of herpes simplex virus. Transport of virus from skin to brainstem nuclei. AB - Herpes simplex virus (HSV) injection into the snout of mice was followed by the appearance of HSV antigen in neurons in trigeminal ganglia, main sensory and spinal tract trigeminal nuclei, reticular formation including raphe nuclei and locus ceruleus on both sides. The findings indicate that HSV spreads via axons, passes through a series of neurons and in this way can reach vital nuclei in the brainstem including monoaminergic neurons from the primary replication area in the lip. PMID- 6281394 TI - Multiple sclerosis. Electrofocused "bands" of oligoclonal CSF IgG do not carry antibody activity against measles, varicella-zoster or rotaviruses. AB - Electrofocused serum and cerebrospinal fluid (CSF) specimens from patients with multiple sclerosis (MS) were analysed for immunoglobulins (Ig) and for antibodies to measles, varicella-zoster and rotaviruses by an imprint immunofixation method. Patterns of intrathecally synthesized antibodies to the 3 viruses differed from patterns of oligoclonal IgG in the CSF. A variable proportion of virus antibody bands (average 19% for measles antibodies, 8% for varicella-zoster antibodies, 31% for rotavirus antibodies) displayed isoelectric points identical to bands of IgG, but absorption with measles, varicella-zoster and rotavirus antigens produced no change in the bands of IgG and no quantifiable decrease of the CSF IgG. The results confirm previous evidence that the intrathecally synthesized viral antibodies so far demonstrated in MS are not carried by the oligoclonal bands of CSF IgG and account for only a minor fraction of the CSF IgG. PMID- 6281395 TI - Mitogenic lectin receptors of nervous system tumors. Study of gliomas, neural crest tumors, and meningiomas in vitro using phytohemagglutinin and concanavalin A. AB - The effects of mitogenic lectins Phytohemagglutinin (PHA), and Concanavalin A (Con A) on the growth rate of cells derived from glial tumors (astrocytoma, ependymoma, glioblastoma, medulloblastoma, and C6 rat glioma), neural crest tumors (neuroblastoma and schwannoma), and meningiomas were studied. The cell lines were of human and animal origin. The specificity of lectin binding to mitogenic receptors was evaluated using complementary monosaccharides. In all glial- and some neural-crest tumor-derived cell lines, there was a lectin concentration-dependent and cell density-dependent, biphasic growth rate response with stimulation at low and inhibition at high lectin concentrations. This response did not depend on the type of glial tumor, species of origin, or passage level in vitro. Although, in meningioma-derived cell lines, lectins did not induce a growth rate response, they caused morphological changes ("whorling"). Lectin stimulation in glial tumor-derived cell lines resembles that occurring in peripheral blood lymphocytes. Lectin-induced mitogenesis may lay the groundwork for the establishment of a model of glial cell proliferation, and that permits the evaluation of cell surface effects, intracellular mechanisms, and epigenetic factors in studies of tumors, neural development, and neuroimmunology. PMID- 6281396 TI - Proton permeability in biological and model membranes. PMID- 6281397 TI - Comparison of pH and calcium dependence of gap junctional conductance. PMID- 6281398 TI - The role of protons in glucose-induced stimulus-secretion coupling in pancreatic islet B cells. PMID- 6281399 TI - Low affinity, adenosine 5'-triphosphate-dependent [3H]dopamine binding in synaptic membranes from bovine caudate is unrelated to dopamine-stimulated adenylate cyclase. PMID- 6281400 TI - Proton nuclear magnetic resonance studies on brain edema. AB - The water in normal and edematous brain tissues of rats was studied by the pulse nuclear magnetic resonance (NMR) technique, measuring the longitudinal relaxation time (T1) and the transverse relaxation time (T2). In the normal brain, T1 and T2 were single components, both shorter than in pure water. Prolongation and separation of T2 into two components, one fast and one slow, were the characteristic findings in brain edema induced by both cold injury and triethyl tin (TET), although some differences between the two types of edema existed in the content of the lesion and in the degree of changes in T1 and T2 values. Quantitative analysis of T1 and T2 values in their time course relating to water content demonstrated that prolongation of T1 referred to the volume of increased water in tissues examined, and that two phases of T2 reflected the distribution and the content of the edema fluid. From the analysis of the slow component of T2 versus water content during edema formation, it was demonstrated that the increase in edema fluid was steady, and its content was constant during formation of TET-induced edema. On the contrary, during the formation of cold-injury edema, water-rich edema fluid increased during the initial few hours, and protein-rich edema fluid increased thereafter. It was concluded that proton NMR relaxation time measurements may provide new understanding in the field of brain edema research. PMID- 6281401 TI - Correlation of methylprednisolone levels in cat spinal cord with its effects on (Na+ + K+)-ATPase, lipid peroxidation, and alpha motor neuron function. AB - Large intravenous doses of methylprednisolone sodium succinate are associated with biochemical and electrophysiological effects in the cat spinal cord which may be of therapeutic value in the treatment of spinal cord injury. The potentially beneficial effects of large doses of the glucocorticoid include: 1) an enhancement of spinal cord (Na + + K+)-ATPase activity; 2) an attenuation of lipid peroxide formation; 3) a hyperpolarization of motor neuron resting membrane potentials; and 4) an accelerated impulse conduction along the myelinated portion of the motor axon. Each of these is apparent with spinal cord tissue levels of methylprednisolone around 1.3 micrograms/gm wet weight, which are rapidly obtained following a single intravenous dose of 30 mg/kg. The half-life of methylprednisolone in cat spinal cord following a single intravenous administration, as well as the duration of its pharmacological actions, is roughly 3 hours. The data suggest that, in order to be of therapeutic value in the treatment of acute spinal cord trauma, early intervention with high-dose intravenous methylprednisolone (30 to 40 mg/kg) is necessary, followed by intravenous maintenance dosing of 15 to 20 mg/kg every 2 to 3 hours. The rationale and duration for this regimen are discussed. PMID- 6281402 TI - Pertechnetate (Tc-99m) sequential scintiphotography--aid in differentiating exudative from transudative pericardial effusions: concise communication. PMID- 6281403 TI - [11C]spiroperidol: synthesis, specific activity determination, and biodistribution in mice. PMID- 6281404 TI - Resting early peak diastolic filling rate: a sensitive index of myocardial dysfunction in patients with coronary artery disease. AB - Resting first-pass radionuclide angiocardiography (RNA) was used to derive left ventricular (LV) peak diastolic filling rates (PFR) in normals (Group 1:N = 12) and in patients with coronary artery disease (CAD), both without (Group 2:N = 27) and with previous myocardial infarction (Group 3:N = 23). Resting peak filling rates were significantly depressed in both Group 2 (1.61 +/- 0.36; p less than 0.01) and Group 3 (1:35 +/- 0.26; p less than 0.001) patients when compared with Group 1, normals (2.14 +/- 0.63). Even though LV systolic function of Group 2 patients was normal and comparable to that in Group 1 (EF = 0.55 +/- 0.06 against EF 0.55 +/- 0.06 NS), diastolic dysfunction [PFR less than 1.61 end diastolic volume/sec (EDV/sec)] was present at rest in 14 of 27 (52%). Depressed PFR values was also seen in 20 of 23 Group 3 patients (87%). It appears that (a) resting PFR is a sensitive and easily obtainable parameter of the diastolic dysfunction associated with CAD; (b) abnormal PFR values are seen in almost all patients with previous myocardial damage, and (c) a significant proportion of CAD patients without any evidence of abnormal systolic function have depressed resting PFR of the LV. PMID- 6281405 TI - Preoperative technetium-99m imaging of a substernal parathyroid adenoma. PMID- 6281406 TI - Brain tumor evaluation using Rb-82 and positron emission tomography. AB - Positron emission tomography of the brain with 75-sec rubidium-82 obtained from a portable generator (25-day Sr-82 leads to Rb-82) was used to evaluate the integrity of the blood-brain barrier (BBB) in patients with brain tumors. Rubidium is normally excluded from the central nervous system by the intact BBB, but when the BBB is disrupted by a tumor. Rb enters and pools in the extravascular spaces of the central nervous system. Since Rb is also rapidly cleared from the blood, a high tissue-to-blood ratio of the Rb-82 tracer is achieved in regions of BBB disruption after intravenous injection. With dynamic positron emission tomographic imaging, the extravasation of the Rb tracer can be evaluated independent of the intravascular Rb concentration, and very small changes in the BBB permeability can be detected. The results of our studies in eight patients show that this technique is a promising method for evaluation of the BBB integrity in brain-tumor patients.U PMID- 6281407 TI - Phosphoenolpyruvate carboxykinase in the small intestine of developing rodents. AB - Phosphoenolpyruvate carboxykinase activity was found to be high in the mucosa of the small intestine of suckling rats. Activity was higher proximally than distally and decreased to very low values at weaning. After weaning, feeding a high fat diet or starvation did not result in elevation of the low intestinal activity in rats. However, in mice, who also showed high activity in the suckling period, starvation, but not diet, did cause a rise. Since fructose-diphosphatase was also found in the intestinal mucosa of suckling rats, it is suggested that gluconeogenesis may occur in this tissue in the neonatal period. PMID- 6281408 TI - Grain dust and alveolar macrophages: an experimental study of the effects of grain dust on the mouse lung. AB - Exposure of mice to grain dust in a dusting chamber for 2-80 days led to a pulmonary macrophage response but no interstitial pulmonary damage. The macrophages contained prominent paracrystalline cytoplasmic inclusions. PMID- 6281409 TI - Concentration of Plasmodium falciparum-infected erythrocytes by density gradient centrifugation in Percoll. PMID- 6281411 TI - Heterogeneity of postsynaptic Alpha adrenergic receptors in mammalian aortas. PMID- 6281410 TI - Modulation of peripheral adrenergic neurotransmission by methionine-enkephalin. AB - The isolated perfusion cat spleen prelabeled with [3H]norepinephrine was used to study the effects of morphine and Met-enkephalin on the exocytotic release of norepinephrine from sympathetic adrenergic neurons after nerve stimulation. The overflow of endogenous norepinephrine, total 3H and dopamine-beta-hydroxylase (DBH) from cat spleens was measured during postganglionic stimulation of the splenic nerve. Perfusion of spleens with Met-enkephalin (10(-8)-10(-5) M) produced a dose-dependent decrease in the release of endogenous norepinephrine upon nerve stimulation. These changes were paralleled by significant dose dependent Met-enkephalin-induced decreases in the nerve stimulation-mediated release of total 3H, DBH and in the perfusion pressure. However, perfusion of spleens with morphine (10(-7)-10(-4) M) produced no significant changes in the release of endogenous norepinephrine, total 3H or DBH after nerve stimulation at 5 Hz. Morphine (10(-7)-10(-4) M) also had no significant effects on the contraction of the splenic capsule. Perfusion of spleens with naloxone (10(-6) M), a pure narcotic antagonist, did not alter the release of endogenous norepinephrine, total 3H, DBH or perfusion pressure. However, perfusion with naloxone (10(-6) M) plus Met-enkephalin (10(-6)-10(-5) M) antagonized the inhibitory effects of Met-enkephalin. These findings support the hypothesis that the opiate receptor population in peripheral tissues are heterogenous and that Met-enkephalin depresses exocytotic release of norepinephrine by interacting with a specific presynaptic opiate receptor. PMID- 6281412 TI - Acute effect of ethanol on prostaglandin E1-mediated cyclic AMP formation by a murine neuroblastoma clone. PMID- 6281413 TI - Distribution of radioactivity in the spinal cord after intracerebroventricular and intravenous injection of radiolabeled opioid peptides in mice. AB - The rejection of [beta-125I]endorphin and D-[3H]Ala2-Met-enkephalinamide into the lateral ventricles of mice revealed that radioactivity could be transported rapidly via the cerebrospinal fluid (CSF) down to the lowest part of the spinal cord. The maximal levels of both compounds in thoracic, lumbar and sacral sections of the spinal cord occurred 10 min after administration, which coincided with the peak antinociceptive action of morphine after i.c.v. administration. When [3H]Leu-enkephalin was administered i.c.v., the level of radioactivity in the spinal cord did not increase with time and these levels in spinal cord were considerably lower than those of either [beta125I]endorphin or D-[3H]Ala2-Met enkephalinamide. This difference in distribution of these compounds, coupled with the observation that [beta-125I] endorphin and D-[3H]Ala2-Met-enkephalinamide were more stable than [3H]Leu-enkephalin in CSF, suggested that radioactivity in the spinal cord of mice treated with [beta-125I]endorphin or D-[3H]Ala2-Met enkephalinamide was due to unchanged material. Very low levels of radioactivity were found in brain or spinal cord after i.v. administration of either [beta 125I]endorphin or D-[3H]Ala2-Met-enkephalinamide. This latter observation supports the view that endorphins administered i.c.v. enter the spinal cord directly through CSF rather than reentry from the general circulation. The results presented herein support the hypothesis that the antinociceptive action of morphine in mice as measured by the tail-flick procedure can be mediated via the release of endogenous compound(s) from brain into CSF which are transported to lower sections of the spinal cord where they inhibit the tail-flick response. PMID- 6281414 TI - Changes in the density of beta adrenergic receptors in rat lymphocytes, heart and lung after chronic treatment with propranolol. AB - Abrupt withdrawal after the chronic administration of propranolol results in clinical syndromes that suggest adrenergic hypersensitivity. Furthermore, propranolol administration has been shown to lead to an increase in the density of beta adrenergic receptors on human lymphocytes. The present studies were designed to assess the relevance of changes measured in lymphocytes to changes that may occur in solid tissues. Direct measurement of the density and properties of beta adrenergic receptors in membrane fragments was performed in vitro using the radioligand [125I]iodohydroxybenzylpindolol. Chronic infusion of propranolol by s.c. implanted osmotic minipumps generated sustained plasma concentrations of propranolol sufficient to cause chronic blockade of beta adrenergic receptors. Infusion of propranolol for 7 days resulted in significant increases in the density of beta adrenergic receptors in rat ventricles, lungs and lymphocytes. A computer-assisted graphic analysis of results obtained in studies with drugs selective for beta-1 or beta-2 receptors revealed increases in the densities of both beta-1 an beta-2 adrenergic receptors. These results are consistent with the hypothesis that change in beta adrenergic receptors on lymphocytes are qualitatively similar to alterations in beta adrenergic receptors in solid tissues not routinely accessible in humans. Increases in the densities of beta-1 and/or beta-2 adrenergic receptors in solid tissues may be related to some of the untoward effects observed in humans after abrupt discontinuation of propranolol administration. PMID- 6281415 TI - Sensitization produced by repeated administration of naloxone is blocked by food deprivation. AB - The current study was conducted in order to explore the effects of repeated naloxone administration as a function of food intake. Rats were trained to press a bar to avoid foot-shock. They were allowed either free or restricted access to food. Free-feeding rats developed a strong sensitivity to naloxone, as manifested by an increased shock rate after naloxone injection. When animals were food deprived, the sensitivity was greatly reduced. A different species (mouse) and two different tests were used to examine further the effects of food intake and pretreatment with naloxone. The mice were given either free access to food or a restricted diet and were pretreated with either naloxone or saline. The effects of food intake and pretreatment with naloxone were examined in terms of motor activity, morphine analgesia and naloxone hyperalgesia. The results showed that prior exposure to naloxone in free-feeding animals enhanced the suppressant effect of naloxone on motor activity and the analgesic effects of morphine (as measured by paw-lick, but not as measured by jump in the hot-plate test), but had no effect on the hyperalgesic effect of naloxone. When mice were food-deprived during naloxone administration, sensitization did not occur. The hypothesis that naloxone sensitivity is due to changes in the number of brain opiate receptors was tested by measuring the number and affinity of [3H]naloxone binding sites on brain membranes from mice chronically treated with naloxone. Neither naloxone pretreatment nor food deprivation affected the number or affinity of binding sites. The gamma-aminobutyric acid antagonist effect of naloxone (as measured by gamma-[3H]aminobutyric acid binding) was also unchanged by naloxone pretreatment. Thus, the basis of the interactions between naloxone and the feeding state remains unclear. PMID- 6281416 TI - Interaction between the renin-angiotensin and beta adrenergic nervous systems in drinking and pressor responses after renal artery constriction. AB - Constriction of the remaining renal artery of a uninephrectomized rat produced an increase in plasma renin level, a decrease in renal cortex renin level, an increase in blood pressure and a drinking response. Simultaneous infusion with the angiotensin II antagonist, saralasin, potentiated the rise in plasma renin level and blocked the rise in blood pressure. Drinking was only partially attenuated. Pretreatment with l-propranolol had no effect on the changes in plasma or kidney renin levels, but the increase in blood pressure was potentiated and the drinking response was attenuated. It is concluded that the pressor and drinking responses to renal artery constriction are partially mediated by the beta adrenergic nervous system. PMID- 6281417 TI - Norepinephrine enhancement of inhibitory synaptic mechanisms in cerebellum and cerebral cortex: mediation by beta adrenergic receptors. PMID- 6281418 TI - Analysis of the negative inotropic effect of acetylcholine on frog atrial fibres. AB - Voltage-clamp experiments have been performed on frog atrial preparations in order to study the mechanism of the inotropic effect of acetylcholine (ACh) at various concentrations. The amplitude of the slow inward current (Is) is reduced even at low ACh concentrations; such low concentrations have little or no effect on potassium permeability. Dose-effect relationships for Is inhibition (Is/Is max) by ACh show a half amplitude dose (K0.5 around 8 X 10(-8) M ACh. The reduction of Is is attributed largely to a decrease of the maximal conductance of the slow channel (gs). Steady-state activation and inactivation parameters are not affected by ACh. Experiments in a Na-free solution (Na replaced by Li ions) or in a Ca-free solution (with EGTA) indicate that the "slow sodium current" is more sensitive to ACh than the "slow Ca current", although these two currents both seem to flow through the slow channel. The decrease of the phasic component of contraction observed in the presence of ACh is very well correlated with the decrease of Is (K0.5 = 8 X 10(-8) M ACh), while the increase of the tonic tension may be related to the outward potassium current induced by high concentrations of ACh. The significant difference between the half amplitude dose (K0.5) observed in the dose effect curves with ACh for Is inhibition (K0.5 = 8 X 10(-8) M) and for ACh-induced extra-current (K0.5 - 10(-6) M) may indicate the presence of two muscarinic receptors. PMID- 6281419 TI - Glucocorticoid binding and control ACTH secretion. AB - This report attempts to summarize the present state of knowledge concerning interaction of glucocorticoids with brain and pituitary receptors, in relation to induction of specific biological effects. It examines the properties of the receptors, emphasizing the fact that transcortin-like binding molecules are present on the cell membrane in the pituitary gland. Also, it considers the functional aspects of the mechanism of action of steroids, including control, occupancy and heterogeneity of binding sites, as well as correlation with regulation of ACTH release. PMID- 6281420 TI - Control of intermediate lobe peptide secretion. AB - It appears that the intermediate lobe of the rat is able to process alpha-MSH, ACTH and endorphins from the precursor pre-opio-corticotrophin (31K). The release of these hormones is under a neural dopaminergic inhibitory control. A beta adrenergic receptor mediating a stimulatory control, also appears to play a role in this release. The possible implications are discussed. PMID- 6281421 TI - Neuroendocrine regulation of stimulated ACTH secretion. AB - Two models of corticosteroid inhibition of stimulus-induced ACTH secretion are discussed. Modulation of ACTH secretion by a nonsteroidal factor related to adrenal mass is reviewed. Experiments are presented that compare feedback inhibition of ACTH affected by corticosterone and by adrenal mass. Use of two stimuli to ACTH secretion and manipulation of two feedback signals allow the conclusion that only one of the steroid feedback models will satisfy the data, and that corticosteroid feedback inhibition of stimulus-induced ACTH secretion must be exerted at at least two sites in the central nervous system. PMID- 6281422 TI - [Hypophyseal corticotropic activity during perinatal period: ontogenesis and regulation]. AB - 1. In the fetal rat hypophysis, the "opio-melano-corticotropic" cells were immunocytologically revealed earlier in the pars distalis (day 16) than in the pars intermedia (day 17). At an early stage of embryonic development, some factors, possibly of nervous origin, could exert inductive influence on the differentiation and/or the increase in the number of immunoreactive cells. 2. Several forms of ACTH, characterized by their apparent molecular weight, bioactivity and immunoreactivity, can be observed when acid extracts are subjected to gel filtration on Sephadex G 50 fine columns. The ratios between these ACTH forms change during fetal development; the rise in bioactive ACTH content during the last days of gestation, is correlated with an increase of the "intermediate" and "little" molecular forms of ACTH and a parallel decrease of the "big" form. 3. The hypophyseal corticotropic activity is highest on days 18 19 of gestation. Between days 17 and 19, the cortico-stimulating activity is largely independent of the hypothalamus although the fetal hypothalamus shows light hypophyso-stimulating influence. In contrast, between days 19 and 21, the hypophyseal corticotropic activity is mostly under the hypothalamic control; the ACTH release is greatly reduced in its absence. 4. Circulating corticosteroids exert a negative feedback directly at the hypothalamic and pituitary levels. Such negative feedback could explain the decrease of the corticotropic activity of the pituitary during the last three days of gestation and the first postnatal days. 5. The fetus at the end of gestation, as the newborn, is responsive to stress; however, the extent of the pituitary response is weaker than in the adult. PMID- 6281423 TI - Neural control of circadian rhythms in plasma ACTH, plasma corticosterone and motor activity. AB - The circadian rhythms for plasma ACTH and corticosterone (B), as well as motor activity, were explored in female rats after ocular enucleation (O-X), stereotaxic lesion of the suprachiasmatic nuclei (SCN-X) or of midbrain raphe nuclei (R-X), or both O-X and R-X, pharmacological blockade of the serotoninergic (5HT) system by pCPA, sometimes bypassed by 5-HTP, or 5-HT denervation of the SCN by local injection of 5,7-DHT. The three circadian rhythms explored responded quite differently to the treatments. In particular, the ACTH and B rhythms lost their usual close correlations. The amplitude and mean level of ACTH fluctuations were depressed after all treatments, but remained normal or were enhanced for B rhythm. ACTH rhythmicity actually was undetectable after SCN-X, pCPA and, in several rats, combined O-X and R-X, whereas persisting circadian and/or ultradian B and locomotor activity rhythms were always measured. The participation of the suprachiasmatic nuclei, the midbrain raphe nuclei and other possible 5-HT components in a complex circadian pacemaker system is discussed. PMID- 6281424 TI - [A study on the physiological basis of training effect-with special reference to myoglobin. II Determination of myoglobin contents in canine skeletal muscles (author's transl)]. PMID- 6281425 TI - Stress reactions and endorphinergic systems. AB - The discovery of the endorphins, a family of distinctive endogenous peptides possessing opiate-like activity, which appear to play a central role in the regulation of pain and other vegetative functions, prompted several investigations in animals and in man as to their possible role in stress phenomena. Several different strategies have been employed: Measurement of concentrations and release of endorphins in different brain areas, measurement of pain sensitivity and of different behavioral variables of animals before and after stress stimuli (with and without the application of the specific opiate antagonist naloxone), measurement of actual levels of endorphins in CSF and plasma of patients with different types of neuroses and psychoses. The present state of research in this field is summarized. PMID- 6281426 TI - High roughage diet and surgical diseases in Afghanistan. PMID- 6281427 TI - Granular cell myoblastoma: a massive breast tumour. PMID- 6281428 TI - Purification and properties of cyclic AMP-dependent protein kinase from rat epididymis. AB - Two protein kinases (I and II: EC 2.7.1.37) that show a high degree of substrate specificity for protamine rather than histones, phosvitin and casein were partly purified from rat epididymal tissue. The enzymes were present in the cytosol because greater than 80% of the enzymic activity was recovered in the soluble fraction. The kinases required Mg2+ for activity although Co2+ and Mn2+ were partial substitutes. Zn2+ (1 mM) inhibited nearly completely the activity of the enzymes. Both the kinases showed high affinity for activation with cyclic AMP compared to other cyclic nucleotides. Amino acid analysis of 32P-labelled protamine product revealed that the kinases transfer the terminal phosphate of ATP to serine residues of the protein. The isoenzymes I and II showed certain differences in relation to their hydroxyapatite-chromatography profiles, pH activation profiles, heat sensitivity and Km for ATP and cyclic AMP. PMID- 6281429 TI - Angiotensin I converting enzyme in rat testis, epididymis and vas deferens under different conditions. AB - Angiotensin I converting enzyme (ACE) was found in the testis, epididymis and vas deferens of rats. In tissue specimens without a prior washout of seminal fluid the highest specific ACE activity was measured in the testis. The enzyme activity was significantly lower towards the end of the excurrent ducts, suggesting that the enzyme is synthesized in the testis and secreted into the seminal fluid there. An ACE inhibiting substance may be secreted by the epididymal epithelium. Enzyme synthesis and enzyme inhibition are probably under simultaneous endocrine control. In-vitro inhibition, pH- and temperature-dependence of gonadal ACE correspond with that of lung and blood plasma. However, the physiological function of the enzyme on sperm motility and fertility remains unsolved. PMID- 6281430 TI - A rapid densitometric microassay for nitroblue tetrazolium reduction and application of the microassay to macrophages. PMID- 6281431 TI - An evaluation of the phagocytic cells in the ia (osteopetrotic) rat: oxidative metabolism in monocytes and macrophages. PMID- 6281432 TI - Differences in the ability of human peripheral blood monocytes and in vitro monocyte-derived macrophages to produce superoxide anion: studies with cells from normals and patients with chronic granulomatous disease. PMID- 6281433 TI - Use of the muscle relaxant atracurium in anephric patients: preliminary communication. AB - Atracurium is a new, non-depolarizing muscle relaxant which rapidly breaks down in vivo and appears to require neither renal nor hepatic function for its elimination. As the commonly used non-depolarizing muscle relaxants carry the risk of prolonged effect when used in anephric patients, atracurium appears to have special promise in this situation. A preliminary account of the successful use of the relaxant in 20 anephric patients is recorded and evidence presented that atracurium is, indeed, a suitable agent for use in patients with renal failure. PMID- 6281434 TI - A family study of Charcot-Marie-Tooth disease. AB - Forty-seven cases of Charcot-Marie-Tooth peripheral neuropathy were seen in 18 families within a defined area, with a disease prevalence of 1 in 16 400. Maximum motor nerve conduction velocity (MNCV) measurement divided off two types of neuropathy (MNCV less than 30 ms-1 and greater than 40 ms-1), but did not distinguish clinically affected from normal in families whose probands had median nerve MNCV greater than 40 ms-1. In the neuronal type of neuropathy ((MNCV greater than 40 ms-1) two genotypes were seen, autosomal dominant (ADN) and autosomal recessive (ARN). Most cases with the demyelinating type (MNCV less than 30 ms-1) had an autosomal dominant genotype (ADD) but one family had possible X linked recessive inheritance (XRD). In one autosomal dominant family a father and son had different electrophysiological types of neuropathy. Peroneal muscle weakness was progressive with age in the ADD genotype and certain patterns of phenotypic features were associated with the major genotypes. Age of onset was not found to be reliable in distinguishing genotypes. Care is needed when counselling isolated male cases because of asymptomatic affected females in the autosomal dominant genotypes, and the possibility of ill defined X linked forms. PMID- 6281436 TI - Physical map of a 470 x 10(3) base-pair region flanking the terminus of DNA replication in the Escherichia coli K12 genome. PMID- 6281437 TI - Relationships between the physical and genetic maps of a 470 x 10(3) base-pair region around the terminus of Escherichia coli K12 DNA replication. PMID- 6281435 TI - Microelectrode studies of the effect of lanthanum on the electrical potential and resistance of outer and inner cell membranes of isolated frog skin. AB - Microelectrodes were used to investigate the effect of 0.5 mM mucosal lanthanum (La3+) on the intracellular potential and the resistance of outer and inner isolated frog skin (Rana esculenta) cell membranes. Under short-circuit conditions, the transapical membrane potential Vsco (mean value = -65.4 +/- 3.2 mV, inside negative) hyperpolarized to -108.7 +/- 2.3 mV in control skins, after addition of the sodium blocker amiloride. Current-voltage curves for the outer and inner membranes were constructed from the amiloride-inhibitable current versus the outer membrane potential Vo or the inner membrane potential Vi. The outer, and to a lesser degree the inner, membrane showed a characteristic nonlinearity with two slope resistances. Addition of La3+ to the outer medium increased the short-circuit current to 190% of the control value. Vsco concomitantly changed to -28 +/- 3.5 mV and outer and inner membrane resistances fell, considerably attenuating the nonlinearity seen in control skins. La3+ is suggested to raise the conductance by its effect on the surface potential. A secondary long-term inhibitory effect of La3+ on short-circuit current has been observed. It is ascribed to the penetration of La3+ into the sodium channels. PMID- 6281438 TI - Physical evidence for early transcription in intracellular bacteriophage P2 DNA. PMID- 6281439 TI - The transposition frequency of IS1-flanked transposons is a function of their size. PMID- 6281440 TI - Structure and stability of Tn9-mediated cointegrates. Evidence for two pathways of transposition. PMID- 6281441 TI - Cleavage of mammalian repetitive deoxyribonucleic acids by a mammalian site specific endodeoxyribonuclease. PMID- 6281442 TI - Transcription in vitro of the bacteriophage P22 antirepressor gene. PMID- 6281443 TI - A derivative of Tn5 with direct terminal repeats can transpose. PMID- 6281444 TI - Regulation of the establishment of repressor synthesis in bacteriophage lambda. PMID- 6281445 TI - Determination of pyrimidine dimer unwinding angle by measurement of DNA electrophoretic mobility. PMID- 6281446 TI - Large deletion mutants of Escherichia coli tRNATyr1. PMID- 6281447 TI - Mild nuclease treatment as a probe for a non-random distribution of adenovirus specific RNA sequences and of cellular RNA in nuclear ribonucleoprotein fibrils. PMID- 6281448 TI - Genomic organization and functional analysis of a deletion variant of the 87A7 heat shock locus of Drosophila melanogaster. PMID- 6281449 TI - Isopropanol and acetone potentiation of carbon tetrachloride-induced hepatotoxicity: single versus repetitive pretreatments in rats. AB - Acute oral pretreatment of rats with isopropanol or acetone results in a dose related potentiation of CCl4 hepatotoxicity. Minimally effective doses (MED) and noneffective doses (NED) of both agents were estimated to be 0.25 and 0.10 ml/kg, respectively. Six MED given twice a day over 3 d caused a greater potentiation than a single MED, but not as much as that produced by the total dose given singly. Six NED given over 3 d did not potentiate CCl4, whereas the total dose did when given singly. A threshold for isopropanol and acetone appears to exist in the rat. A total dose of 1.5 ml/kg acetone was administered by four different treatment regimens (bolus, divided doses, infusion) over 3 d. Potentiation of CCl4 hepatotoxicity was then correlated with blood pharmacokinetic parameters: area under the concentration-time curve and peak blood concentration. An excellent correlation was found between the degree of potentiation observed and the peak blood concentration attained, but no correlation was found with area under the curve. Results of iv acetone infusions (over 3 d) with higher doses support the hypothesis that a threshold concentration of acetone is critical in the potentiation of CCl4 hepatotoxicity in the rat. PMID- 6281450 TI - Volcanic ash: toxicity to isolated lung cells. AB - Samples of volcanic ash from Mount St. Helens were collected from Spokane, Washington, after the major eruption of May 18, 1980. The toxicity of ash to the lung was estimated by monitoring the effects of in vitro and in vivo exposure on various physiological parameters of isolated lung cells. Volcanic ash had little effect on O2 consumption of rabbit type II pneumocytes, O2 consumption or superoxide release of resting rat alveolar macrophages, or membrane integrity of rat alveolar macrophages. Ash also caused no significant lipid peroxidation in rat lung microsomes. However, volcanic ash did inhibit superoxide anion release from zymosan-stimulated rat alveolar macrophages. Since superoxide is an antibacterial substance, this result suggests that exposure to volcanic ash may adversely affect the ability of alveolar macrophages to protect the lung from infection. PMID- 6281451 TI - Cortisol and corticotrophin in burned patients. AB - In a study of 36 men burned in a fire, based on sequential early morning samples, plasma cortisol concentration was elevated in proportion to burn size. Plasma corticotrophin (ACTH) was not correlated with burn size, suggesting that factors other than ACTH contribute to the elevated cortisol. Cortisol levels did not fall on the days preceding death in nonsurvivors. During 24-hr sampling, burned patients exhibited a fitted cortisol curve mean that was elevated in proportion to burn size, a rhythm amplitude that was significantly less than that in uninjured controls, and a normal peak time. Metabolic rate, rectal temperature, and urinary catecholamine excretion were also elevated in proportion to burn size. Although plasma cortisol was positively correlated with metabolic rate and with temperature, this appeared to result from a common relationship of these variables with burn size. On the other hand, urinary catecholamine values significantly reduced the residual variance of metabolic rate and temperature after accounting for variance related to burn size. Cortisol appears to be less prominent than catecholamines as a possible mediator of the elevated thermogenesis. PMID- 6281452 TI - Radionuclide salivary gland imaging of maxillary sinus oncocytoma. PMID- 6281453 TI - Dark-field electron microscopy of small peptide hormones: adrenocorticotrophic hormone, glucagon, secretin. PMID- 6281454 TI - In vitro studies of the mechanism of leukemogenesis. II. Characterization of endogenous murine leukemia viruses isolated from AKR thymic epithelial reticulum cell lines. AB - Thymic epithelial reticulum (TER) cell lines were established from thymuses of a young healthy AKR mouse (A2T), a preleukemic AKR mouse (A6T), and two lymphoma bearing AKR/Ms mice (ASLT-1 and ASLT-2). Numerous type-C virus particles with occasional budding forms were observed in all cell lines. Expression of XC detectable, N-tropic, ecotropic virus was observed in every cell line, whereas the presence of xenotropic and mink cell focus-inducing (MCF) viruses could be detected only in TER cells derived from preleukemic and leukemic mice. Expression of xenotropic virus in various cells of newborn and young AKR mice could readily be induced by IUdR treatment, whereas MCF virus was never detected in these cells, with the exception of the A2T cell line after more than 20 passages, in which MCF virus with dual-tropic infectivity emerged in addition to ecotropic and xenotropic viruses. These spontaneous and induced MCF viruses were purified, and their virological properties were characterized. The cloned MCF viruses (MCFs AT1, AT2, AT3, and AT4-IU) showed dual tropism and produced cytopathic effect like foci in mink lung cells. Preinfection with either ecotropic or xenotropic virus interfered with the infectivity of MCF viruses. Spontaneous leukemogenesis in AKR mice was accelerated by the inoculation of MCF viruses. These findings indicate that TER cells could serve as the host cells for the genetic recombination of the endogenous MuLV; the recombinant MuLV, MCF virus, appears to be most closely associated with leukemogenesis in AKR mice. PMID- 6281455 TI - Characterization of the major Epstein-Barr virus-specific RNA in Burkitt lymphoma derived cells. AB - Cytoplasmic RNA prepared from five lymphoid cell lines and a Burkitt lymphoma biopsy was radioactively labeled in vitro and hybridized to cloned EcoRI restriction endonuclease fragments of B95-8 Epstein-Barr virus DNA. The results confirmed that the most abundant cytoplasmic RNA species in such cells is specified by a small region of the genome defined by the EcoRI J fragment. Detailed mapping experiments precisely localized these transcripts within the sequence of the rightmost one-third of the EcoRI J fragment. DNA sequencing suggested that this region of the Epstein-Barr virus genome is unable to code for protein. The major early transcripts consisted of two non-polyadenylated RNA species, each about 170 nucleotides in length. They were both transcribed off the same strand of the DNA and showed significant sequence homology with each other. The coding sequences of the two small RNAs contained potential intragenic control regions for RNA polymerase III. PMID- 6281456 TI - Physical mapping of adenovirus type 2 temperature-sensitive mutations by restriction endonuclease analysis of interserotypic recombinants. AB - The genome structures of about 100 interserotypic ts recombinants produced in crosses between human adenovirus type 2 (H2) and 5 (H5) temperature-sensitive mutants were analyzed by cleavage with restriction endonucleases to determine the map coordinates of the following temperature-sensitive mutants: penton base plus fiber-defective H2 ts103, -104, and -136, assembly-defective H2 ts112, fiber defective H2 ts125, hexon-defective H2 ts118 and -121, and DNA-negative H2 ts111. H5 ts1 (100 K defective), H5 ts36 (DNA negative), H5 ts125 (mutated in the early 72,000-dalton protein), H5 ts22 (fiber defective), H5 ts58 (IIIa defective), and H5 ts18 and -19 were used as one of the parents. The physical locations of the H2 temperature-sensitive mutations thus defined are discussed in relation to the genetic map, the biological function altered, and the positions of the structural genes on the genome. PMID- 6281457 TI - Processing and amino acid sequence analysis of the mouse mammary tumor virus env gene product. AB - The envelope proteins of mouse mammary tumor virus (MMTV) are synthesized from a subgenomic 24S mRNA as a 75,000-dalton glycosylated precursor polyprotein which is eventually processed to the mature glycoproteins gp52 and gp36. In vivo synthesis of this env precursor in the presence of the core glycosylation inhibitor tunicamycin yielded a precursor of approximately 61,000 daltons (P61env). However, a 67,000-dalton protein (P67env) was obtained from cell-free translation with the MMTV 24S mRNA as the template. To determine whether the portion of the protein cleaved from P67env to give P61env was removed from the NH2-terminal end of P67env and as such would represent a leader sequence, the NH2 terminal amino acid sequence of the terminal peptide gp52 was determined. Glutamic acid, and not methionine, was found to be the amino-terminal residue of gp52, indicating that the cleaved portion was derived from the NH2-terminal end of P67env. The NH2-terminal amino acid sequences of gp52's from endogenous and exogenous C3H MMTVs were determined though 46 residues and found to be identical. However, amino acid composition and type-specific gp52 radioimmunoassays from MMTVs grown in heterologous cells indicated primary structure differences between gp52's of the two viruses. The nucleic acid sequence of cloned MMTV DNA fragments (J. Majors and H. E. Varmus, personal communication) in conjunction with the NH2 terminal sequence of gp52 allowed localization of the env gene in the MMTV genome. Nucleotides coding for the NH2 terminus of gp52 begin approximately 0.8 kilobase to the 3' side of the single EcoRI cleavage site. Localization of the env gene at that point agrees with the proposed gene order -gag-pol-env- and also allows sufficient coding potential for the glycoprotein precursor without extending into the long terminal repeat. PMID- 6281458 TI - Proteins containing only half of the coding information of early region 1b of adenovirus are functional in human cells transformed with the herpes simplex virus type 1 thymidine kinase gene and adenovirus type 2 DNA. AB - We introduced into tk- human 143 cells adenovirus type 2 (Ad2) genes by transformation with a plasmid (p711) containing both Ad2 sequences and the herpes simplex virus type 1 (HSV-1) tk gene. p711 contained approximately the left 8% of the Ad2 genome inserted in the HindIII site of pBR322, whereas the fragment of HSV-1 containing the tk gene was inserted in the BamHI site. Three tk+ cell lines were isolated after selection in HAT medium. The arrangement of viral sequences in the three transformants was analyzed by restriction endonuclease digestion and filter hybridization. All three lines contained a single insertion of Ad2 DNA which was present at approximately one copy per cell. The arrangement of Ad2 sequences in these lines was identical to that found in the linear p711 DNA used in the transformation. S1 analysis of the Ad2-specified RNA from two of these lines indicated that the early region 1a mRNA's were synthesized, though in lower amounts than found in lytic infections. These cell lines contained only the left half of early region 1b (4.6 to 11.2), which encoded the 5' portion of the 1b mRNA's. A complex pattern of 1b RNAs was made in these cell lines. Transcription of most of these RNAs began at or near the 1b promoter and proceeded through the 1b sequences into the flanking pBR322, HSV-1, or host sequences. Since many of the RNAs were terminated or spliced in the HSV-1 (anti-sense strand) or pBR322 sequences, new RNA processing sites must be used in the formation of these mRNA's. All three lines fully complemented the 1a deletion mutant Ad5 dl312. Surprisingly, these lines also permitted the growth of 1b deletion mutants (Ad5 dl313 and Ad5 dl434), although the complementation was not always complete. Presumably the new gene product(s) which contained only part of the 1b gene provided most of the essential function(s) required for viral multiplication. Alternatively, the 1b 19-kilodalton protein which was entirely encoded by the adenovirus sequences present in these cell lines was sufficient for viral growth even in the absence of the 1b 55-kilodalton protein. PMID- 6281459 TI - Characterization of proviruses cloned from mink cell focus-forming virus-infected cellular DNA. AB - Two proviruses were cloned from EcoRI-digested DNA extracted from mink cells chronically infected with AKR mink cell focus-forming (MCF) 247 murine leukemia virus (MuLV), using a lambda phage host vector system. One cloned MuLV DNA fragment (designated MCF 1) contained sequences extending 6.8 kilobases from an EcoRI restriction site in the 5' long terminal repeat (LTR) to an EcoRI site located in the envelope (env) region and was indistinguishable by restriction endonuclease mapping for 5.1 kilobases (except for the EcoRI site in the LTR) from the 5' end of AKR ecotropic proviral DNA. The DNA segment extending from 5.1 to 6.8 kilobases contained several restriction sites that were not present in the AKR ecotropic provirus. A 0.5-kilobase DNA segment located at the 3' end of MCF 1 DNA contained sequences which hybridized to a xenotropic env-specific DNA probe but not to labeled ecotropic env-specific DNA. This dual character of MCF 1 proviral DNA was also confirmed by analyzing heteroduplex molecules by electron microscopy. The second cloned proviral DNA (designated MCF 2) was a 6.9-kilobase EcoRI DNA fragment which contained LTR sequences at each end and a 2.0-kilobase deletion encompassing most of the env region. The MCF 2 proviral DNA proved to be a useful reagent for detecting LTRs electron microscopically due to the presence of nonoverlapping, terminally located LTR sequences which effected its circularization with DNAs containing homologous LTR sequences. Nucleotide sequence analysis demonstrated the presence of a 104-base-pair direct repeat in the LTR of MCF 2 DNA. In contrast, only a single copy of the reiterated component of the direct repeat was present in MCF 1 DNA. PMID- 6281460 TI - Initiation and regulation of simian virus 40 early transcription in vitro. AB - We have studied initiation and regulation of early transcription of simian virus (SV40) DNA in vitro by eucaryotic RNA polymerase II, using both a crude HeLa cell extract and a partially purified calf thymus polymerase supplemented with a HeLa cell S100 fraction. Analysis of initiation sites by primer-directed cDNA synthesis and sequencing of cDNA's has revealed that early transcription is initiated at a multiplicity of sites corresponding to the 5' termini of early viral mRNA's. The pattern of in vitro initiation closely resembles the pattern of 5' termini of early mRNA's late in the lytic cycle, with principal initiations between residues 5184 to 5194, upstream from the early Hogness-Goldberg (TATA) sequence, and at residue 5123, well downstream from this sequence. In vitro transcription is initiated to a lesser extent at sites between residues 5150 and 5155, the principal 5' termini of early mRNA's in transformed cells and early in lytic infection, located 21 to 26 nucleotides downstream from the TATA sequence. Initiation occurs at identical sites and with similar efficiencies on form I and linearized DNA templates. There are minor differences in the efficiency of initiation at specific sites by the two transcriptional systems. Studies using a DNA template cleaved just downstream from the TATA sequence and a second template cleaved through a pair of 72-base-pair tandem repeats starting 87 nucleotides upstream from the TATA sequence have revealed that neither the TATA sequence nor the repeats are essential for early transcription in vitro. However, removal of the TATA and upstream sequences shifts initiation of transcription principally to the residue 5123 site. Comparison of the relative efficiencies of transcription on intact wild-type DNA, the two cleaved DNAs, and DNA from a deletion mutant suggests that all or most of the sequences constituting an early promoter lie within the genomic region 60-70 to 140 nucleotides upstream from the principal 5' termini of the early mRNA's. PMID- 6281461 TI - Temporal patterns of human cytomegalovirus transcription: mapping the viral RNAs synthesized at immediate early, early, and late times after infection. AB - The transcription of the human cytomegalovirus genome was investigated at immediate early, early, and late times after infection. Viral RNAs associated with either the whole cell, the nucleus, the cytoplasm, or the polyribosomes were analyzed. At immediate early times, i.e., in the absence of de novo viral protein synthesis, the viral RNA in high abundance originated from a region of the long unique section of the prototype arrangement of the viral genome (0.660 to 0.770 map units). The viral RNA in low abundance originated from the long repeat sequences (0.010 to 0.035 and 0.795 to 0.825 map units) and a region in the long unique section (0.201 to 0.260 map units). Viral RNAs associated with the polyribosomes as polyadenylated RNA were mapped to these restricted regions of the viral genome and characterized according to size class in kilobases. At 24 h after infection in the presence of an inhibitor of viral DNA replication, i.e., at early times, the stable viral RNAs in highest abundance mapped in the long repeat sequences. Viral RNAs at intermediate abundance under these conditions mapped in two regions of the long unique section of the viral genome (0.325 to 0.460 and 0.685 to 0.770 map units). Stable viral RNAs that were associated with the polyribosomes in high abundance as polyadenylated RNA orginated from the long repeat sequences, but not from the long unique section of the viral genome. An analysis of whole-cell RNA at late times (72 h) indicated that the abundant transcription was in the regions of the long unique sequences (0.325 to 0.460 and 0.660 to 0.685 map units), and transcription of intermediate abundance was from the long repeat sequences. However, stable viral mRNA's derived from the long repeat sequences were associated with the polyribosomes at late times after infection. In addition, mRNA's originating from the long and short unique sequences were found associated with the polyribosomes at higher relative concentration than at early times after infection. It is proposed that expression of the immediate early viral genes is required to transcribe the early viral genes in the long repeat and adjacent sequences. These sequences are also transcribed at late times after infection while viral DNA synthesis continues. The expression of viral genes in most of the long and short unique sequences appears to require viral DNA replication. PMID- 6281462 TI - McDonough feline sarcoma virus: characterization of the molecularly cloned provirus and its feline oncogene (v-fms). AB - The genetic structure of the McDonough strain of feline sarcoma virus (SM-FeSV) was deduced by analysis of molecularly cloned, transforming proviral DNA. The 8.2 kilobase pair SM-FeSV provirus is longer than those of other feline sarcoma viruses and contains a transforming gene (v-fms) flanked by sequences derived from feline leukemia virus. The order of genes with respect to viral RNA is 5' gag-fms-env-3', in which the entire feline leukemia virus env gene and an almost complete gag sequence are represented. Transfection of NIH/3T3 cells with cloned SM-FeSV proviral DNA induced foci of morphologically transformed cells which expressed SM-FeSV gene products and contained rescuable sarcoma viral genomes. Cells transformed by viral infection or after transfection with cloned proviral DNA expressed the polyprotein (P170gag-fms) characteristic of the SM-FeSV strain. Two proteolytic cleavage products (P120fms and pp55gag) were also found in immunoprecipitates from metabolically labeled, transformed cells. An additional polypeptide, detected at comparatively low levels in SM-FeSV transformants, was indistinguishable in size and antigenicity from the envelope precursor (gPr85env) of feline leukemia virus. The complexity of the v-fms gene (3.1 +/- 0.3 kilobase pairs) is approximately twofold greater than the viral oncogene sequences (v-fes) of Snyder-Theilen and Gardner-Arnstein FeSV. By heteroduplex, restriction enzyme, and nucleic acid hybridization analyses, v-fms and v-fes sequences showed no detectable homology to one another. Radiolabeled DNA fragments representing portions of the two viral oncogenes hybridized to different EcoRI and HindIII fragments of normal cat cellular DNA. Cellular sequences related to v-fms (designated c-fms) were much more complex than c-fes and were distributed segmentally over more than 40 kilobase pairs in cat DNA. Comparative structural studies of the molecularly cloned proviruses of Synder-Theilen, Gardner-Arnstein, and SM-FeSV showed that a region of the feline-leukemia virus genome derived from the pol-env junction is represented adjacent to v-onc sequences in each FeSV strain and may have provided sequences preferred for recombination with cellular genes. PMID- 6281463 TI - Mapping of the 5' termini of two adeno-associated virus 2 RNAs in the left half of the genome. AB - The left 45% of the adeno-associated virus 2 genome was sequenced. The 5' termini of two adeno-associated virus-specific RNAs were mapped at the nucleotide level within this region of the genome (nucleotides 286 to 288 and 871 to 874). Both of these 5' termini map 31 +/- 2 nucleotides downstream from the start of "TATA' boxes. PMID- 6281464 TI - The leader sequence of the subgenomic mRNA's of Rous sarcoma virus is approximately 390 nucleotides. AB - The subgenomic mRNA's of Rous sarcoma virus share a common 5' leader sequence spliced from genomic RNA. We have examined the 5' terminal sequences of four Rous sarcoma virus RNAs: virion RNA and three species of intracellular mRNA which direct the synthesis of the RSV gene products. The lengths of the leaders on the RNAs were determined by the extent that they could protect cloned Rous sarcoma virus DNA fragments from S1 nuclease digestion after RNA-DNA hybridization. We found that the subgenomic mRNA's that direct the synthesis of the env and src gene products have uninterrupted spliced leader sequences of approximately 390 nucleotides, whereas virion RNA and full-length intracellular viral RNA have 5' termini homologous to the cloned viral DNA probe over at least the first 735 bases. In the accompanying manuscript we have determined the nucleotide sequence of the 5' end of the Rous sarcoma virus genome, including the candidate splice donor site identified here (Swanstrom et al., J. Virol. 41:535-541, 1982). PMID- 6281465 TI - Nucleotide sequence of the 5' noncoding region and part of the gag gene of Rous sarcoma virus. AB - Several functions of the retrovirus genome involve structural features in the vicinity of its 5' terminus. In an effort to further elucidate the relationship between structure and function in retrovirus RNA, we have determined the sequence of the first 1,010 nucleotides at the 5' end of the genome of Rous sarcoma virus by using the Maxam-Gilbert method to sequence suitable domains in cloned Rous sarcoma virus DNA. The results (i) locate the initiation codon for the gag gene of Rous sarcoma virus 372 nucleotides from the 5' end of viral RNA; (ii) demonstrate that this codon is preceded by three methionine codons that are apparently not used in translation; (iii) sustain previous conclusions that the principal site to which ribosomes bind on the Rous sarcoma virus genome in vitro does not contain the initiation codon for gag; (iv) permit deduction of the amino acid sequence of a viral structural protein, p19; (v) confirm the amino-terminal sequence of Pr76gag; and (vi) substantiate the identification of a splice donor site described in the accompanying manuscript (Hackett et al., J. Virol., 41:527 534, 1982). PMID- 6281466 TI - Long terminal repeat of murine retroviral DNAs: sequence analysis, host-proviral junctions, and preintegration site. AB - The nucleotide sequence of the long terminal repeat (LTR) of three murine retroviral DNAs has been determined. The data indicate that the U5 region (sequences originating from the 5' end of the genome) of various LTRs is more conserved than the U3 region (sequences from the 3' end of the genome). The location and sequence of the control elements such as the 5' cap, "TATA-like" sequences, "CCAAT-box," and presumptive polyadenylic acid addition signal AATAAA in the various LTRs are nearly identical. Some murine retroviral DNAs contain a duplication of sequences within the LTR ranging in size from 58 to 100 base pairs. A variant of molecularly cloned Moloney murine sarcoma virus DNA in which one of the two LTRs integrated into the viral DNA was also analyzed. A 4-base pair duplication was generated at the site of integration of LTR in the viral DNA. The host-viral junction of two molecularly cloned AKR-murine leukemia virus DNAs (clones 623 and 614) was determined. In the case of AKR-623 DNA, a 3- or 4 base-pair direct repeat of cellular sequences flanking the viral DNA was observed. However, AKR-614 DNA contained a 5-base-pair repeat of cellular sequences. The nucleotide sequence of the preintegration site of AKR-623 DNA revealed that the cellular sequences duplicated during integration are present only once. Finally, a striking homology between the sequences flanking the preintegration site and viral LTRs was observed. PMID- 6281467 TI - Further characterization of mRNA's of mouse hepatitis virus: presence of common 5'-end nucleotides. AB - The mouse hepatitis virus strain A59 codes for seven RNA species in the infected cells. These virus-specific RNAs were found to be polysome associated and therefore likely to represent mRNA's. All of them have common 3'-end sequences (Lai et al., J. Virol. 39:823-834, 1981). Their structure was further studied with respect to their 5'-end sequences. It was found that all of these mRNA's contained cap structures at their 5' ends. Furthermore, the cap-containing oligonucleotides which represent the sequences immediately adjacent to the 5' ends were found to be the same for most, if not all, of the seven virus-specific mRNA's. These sequences are also identical to the 5'-end sequences of the virion RNA genome. The 5'-end sequences were tentatively determined to be 5'-cap-N-UAAG. The presence of the common nucleotides in all of the virus-specific RNAs in mouse hepatitis virus strain A59 suggests several possible mechanisms of synthesis for these RNAs. The significance of these findings is discussed. PMID- 6281468 TI - Isolation of vesicular stomatitis virus defective interfering genomes with different amounts of 5'-terminal complementarity. AB - I isolated at least 30 different vesicular stomatitis virus defective interfering (DI) genomes, distinguished by chain length, by five independent undiluted passages of a repeatedly cloned virus plaque. Labeling of the 3' hydroxyl ends of these DI genomes and RNase digestion studies demonstrated that the ends of these DI genomes were terminally complementary to different extents (approximately 46 to 200 nucleotides). Mapping studies showed that the complementary ends of all of the DI genomes were derived from the 5' ends of the nondefective minus-strand genome. Regardless of the extent of terminal complementarity, all of the DI genomes synthesized the same 46-nucleotide minus-strand leader RNA. PMID- 6281469 TI - Organization of type C viral DNA sequences endogenous to baboons: analysis with cloned viral DNA. AB - Unintegrated linear and circular forms of baboon endogenous type C virus M7 DNA were prepared from M7-infected cells by chromatography on hydroxyapatite columns, and the circular DNAs were purified in cesium chloride-ethidium bromide equilibrium density gradients. The circular DNAs were linearized by digestion with EcoRI, which had a unique site on the viral DNA. The linearized DNA was then inserted into lambda gtWES. lambda B at the EcoRI site and cloned in an approved EK2 host. Molecularly cloned full-length M7 DNA was restricted with BamHI, and the resulting five subgenomic fragments were then subcloned individually in plasmid pBR322. The organization and sites of integration of the approximately 100 copies of M7 DNA sequences endogenous to baboons were investigated by digesting the DNA with restriction enzymes and identifying the virus-specific fragments by hybridization to labeled probes made by using the molecularly cloned full-length and subgenomic fragments of the viral DNA. We found that most of the endogenous sequences had sizes and organizations similar to those of the unintegrated viral DNA and therefore approximately similar to the RNA of the infectious virus. A few of the multiple sequences had deletions in the 3' end (envelope region), and some of the sequences either lacked or contained modified BamHI restriction sites on the 5' end of the viral DNA. The endogenous viral DNA sequences were nontandem, uninterrupted, and colinear with the DNA of the infectious virus, and they were integrated at different sites in the baboon DNA, like the M7 proviral DNA sequences acquired upon infection. PMID- 6281470 TI - Deletions of specific regions of the simian sarcoma-associated virus genome are found in defective viruses and in the simian sarcoma virus. AB - Extrachromosomal DNA was isolated from tissue culture cells that were acutely infected with simian sarcoma virus (SSV) and its associated helper (simian sarcoma-associated virus [SSAV]). Two sizes of closed circular viral genomic DNA intermediates were isolated, cleaved at the single EcoRI site, and ligated to the Charon 21A phage lambda vector. Cloned molecules of the larger size all represented the full-length (9.0-kilobase [kb]) SSAV molecule. A heterogeneous group of clones was derived from the smaller DNA circles. These included the SSV genome and SSAV deletion mutants. When two SSV clones were compared with the helper, they contained the following three characteristic deletions: (i) a 250 base pair deletion in the gag gene about 1.0 kb from the 5' end of the genome; (ii) a 1.85-kb deletion in the pol gene; and (iii) a 1.9-kb deletion at the 3' end, which included part of the env gene. This latter deletion was the site of the onc gene substitution. Six other clones of the smaller molecules represented the following variants of the SSAV genome: (i) two clones of the entire genome containing only one long terminal repeat unit; (ii) one clone with the 1.85-kb deletion of the pol gene observed in SSSV; and (iii) three clones having a deletion of the 3' end of the SSAV genome. In each of the latter clones, the 5' border of the deletion was indistinguishable from the 5' border of the onc substitution in SSV. The fidelity of genetic deletions observed suggested that certain regions of the SSAV genome were deleted at a high frequency. In certain cases, these deletions may have been accompanied by a substitution of cellular sequences to generate SSV. PMID- 6281471 TI - Characterization of genome structure of amphotropic and ecotropic wild mouse retroviruses. AB - We studied the RNA genomes of several wild mouse type C retroviruses by using RNase T1-oligonucleotide fingerprinting. The amphotropic and ecotropic viruses of field strain 1504 produced very similar oligonucleotide fingerprints, but each also had several unique oligonucleotides. All of these unique oligonucleotides were located in the env gene region and were probably responsible for the host range differences between these viruses, as well as the lymphomagenic and paralytogenic properties of the viruses. We obtained similar results with the amphotropic and ecotropic viruses of another field strain (4070), which was isolated from a mouse from a different trapping area. The amphotropic viruses of several field strains (strains 1504, 4070, and 1313) were more closely related than the ecotropic viruses of different strains (strains 1504, 4070, and 4996). These findings suggested that the genetic sequences of the amphotropic viruses are more conserved than those of ecotropic viruses isolated from the same wild mice. PMID- 6281472 TI - Guanidine-resistant defective interfering particles of poliovirus. AB - A mixture containing standard poliovirus and D3 particles (mutants with deletions in the capsid locus) was serially passaged in the presence of guanidine. Within five growth cycles, the standard virus was guanidine resistant, but the D3 particles were guanidine sensitive, even after 21 passages with the inhibitor. By passage 40 with guanidine, D3 particles were eliminated, and a new deletion mutant (DX) appeared in the virus population. D3 particles contained a 15% deletion, and DX particles contained a 6% deletion in the capsid locus. Although neither mutant induced the synthesis of NCVP1a or a complete complement of capsid proteins after infection, cells infected with DX particles produced two novel proteins, which had molecular weights of approximately 68,000 and 25,000. PMID- 6281473 TI - Molecular properties of a gag- pol- env+ murine leukemia virus from cultured AKR lymphoma cells. AB - We have described the isolation of a replication-defective murine leukemia virus from a culture of AKR lymphoma cells [Rein et al., Nature (London) 282:753-754, 1979]. To facilitate the characterization of this murine leukemia virus, we transmitted it to mink cells and analyzed its genome by restriction mapping of the mink cellular DNA. This genome resembled the Akv genome quite closely, but it had an additional KpnI cleavage site at 1.3 kilobase pairs from the 5' end of the provirus and a small (approximately 50-base-pair) deletion between 1.8 and 3.0 kilobase pairs from the 5' end. When we tested these mink cells by immune precipitation or by competition radioimmunoassay, we found that they synthesized gPr82env, but contained no detectable gag or pol proteins. It seems likely that the KpnI cleavage site at 1.3 kilobase pairs reflects an abnormal sequence at or near the beginning of the gag gene, which prevents gag or pol translation by introducing a frameshift or termination codon into this region. PMID- 6281474 TI - Structural relationship between a normal chicken DNA locus and the transforming gene of the avian acute leukemia virus MC29. AB - We screened a recombinant chicken DNA/lambda phage library for sequences homologous to the transformation-specific sequences of the avian acute leukemia virus MC29 by hybridization with molecularly cloned MC29 proviral DNA. Three cellular DNA clones were found and compared with each other and with the viral genome by physical mapping with restriction endonucleases and by heteroduplex analysis. These experiments indicated that the three cellular clones overlap and represent a single cellular locus. The RNA genome of MC29 and normal cell DNA share a homologous region of 1.6 kilobases which is interrupted in the cellular DNA by 1.0 kilobase of sequences not present in the viral genome. Hybridization of the cloned cellular DNA to viral RNA and analysis of the protected viral RNA by fingerprinting techniques indicated that there is extensive sequence homology between the helper virus-unrelated mcv sequences of the viral RNA and the cellular DNA, with only minor base differences. The cellular mcv locus, however, lacks all helper virus-related sequences of MC29, including those of the partial viral gag gene which, together with mcv, encodes the probable transforming protein of MC29. We conclude that although the mcv locus of the normal cell does not represent a complete structural homolog to the onc gene of MC29, it is probably the precursor to the onc-specific sequence in the virus. PMID- 6281475 TI - Use of fluoresceinated Epstein-Barr virus to study Epstein-Barr virus-lymphoid cell interactions. AB - As a direct approach to visualize Epstein-Barr virus (EBV) binding to its cellular receptors and to learn more on the nature of this binding, virus preparations were conjugated to fluorescein isothiocyanate and used to detect EBV receptors on lymphoid cells. Different enzymatic and chemical treatments were also applied either to the virus or to target cells or to both, and the effect of these treatments on virus binding was then examined. The results obtained show that: (i) EBV can be fluoresceinated without affecting its infectivity or cell binding ability, and the fluoresceinated virus represents an important tool to investigate the biology and nature of EBV interactions with its cellular receptors; (ii) the two virus strains (P3HR-1 and B95-8) share common receptors on Raji cells; (iii) protease treatment of EBV or target cells abrogates virus binding; (iv) EBV receptors regenerate after removal of the protease, and this regeneration is inhibited by cycloheximide or sucrose; (v) EBV particles bear concanavalin A receptors, and this lectin hinders the interaction of the virus with its cellular receptors; (vi) neuraminidase treatment, various monosaccharides, ovalbumin, and glycopeptides derived from EBV or cell surface do not inhibit virus binding. Taken together, the above data also demonstrate that some cellular and viral surface (glyco-) proteins are required for EBV binding to its targets. PMID- 6281476 TI - Involvement of the bacterial groM gene product in bacteriophage T7 reproduction. I. Arrest at the level of DNA packaging. AB - The multiplication of bacteriophage T7 is blocked in Escherichia coli M. The genetic determinant of this ability (groM) to inhibit T7 growth was transferred to an E. coli K-12 recipient by means of conjugation. We determined at which precise step T7 maturation is blocked. Phage-directed protein and DNA synthesis as well as degradation of host DNA were not qualitatively affected. Instead of infective phages, only preheads were produced. These, however, were maturable in vitro. The newly synthesized phage DNA accumulated in a concatemeric form and matured from its tetrameric or longer forms (very fast sedimenting DNA) only into its dimeric form (fast-sedimenting DNA) or longer forms. The following step, i.e., the maturation of the dimeric to unit-length DNA, was not observed. Since the concatemeric form of T7 DNA accumulated in spite of the presence of maturable preheads, it is likely that the maturation process was blocked at the level of DNA packaging. As intermediates in the packaging process, we found some prehead DNA complexes. We interpreted these as true assembly intermediates (or breakdown products thereof), since the attached DNA was still in its concatemeric form. This shows that the very first DNA packaging step, the binding of the progeny DNA to the preheads, was obviously not blocked. Rather, a later step, such as the filling of the preheads with T7 DNA or the stabilization of completely packaged particles (i.e., the final cutting of the concatemers into unit-size length), was inhibited. PMID- 6281477 TI - Limited transcription of the herpes simplex virus genome when latent in human sensory ganglia. AB - Human paravertebral ganglia were examined for the presence of specific sequences of viral RNA by in situ cytological hybridization, using as probes individual fragments of herpes simplex virus type 2 DNA. Ganglia were obtained at autopsy from 40 patients, and 14 were positive for herpes simplex virus RNA. Transcripts from the left-hand 30% of the viral genome were detected in all of the herpes simplex virus-positive cases. RNA homologous to other sequences from the L component were present less frequently, and no RNA from the S component was detected. These results argue that specific transcription of the herpes simplex virus genome occurs in latently infected human ganglion cells. PMID- 6281478 TI - Inhibition of Sindbis virus maturation after treatment of infected cells with trypsin. AB - Brief treatment of Sindbis virus-infected BHK-21 or Vero cells with low concentrations of trypsin irreversibly blocked further production of progeny virions after removal of the enzyme. The inhibitory effects of the trypsin treatment could only be demonstrated in cells in which virus infection was established; optimal inhibition occurred at ca. 3 h postinfection. Production of virus structural proteins PE2, E1, and C occurred at normal levels in inhibited cells. PE2 and E1 were also transported to the cell plasma membrane during inhibition; however, PE2 was not cleaved to E2, and little capsid protein became membrane associated relative to control cells. Although trypsin treatment had no effect on Sindbis protein synthesis, the production of both 26S and 42S RNA was greatly reduced. Similar trypsin treatment of BHK cells infected with vesicular stomatitis virus had no detectable effect on the course of virus infection. PMID- 6281479 TI - Induction of Epstein-Barr virus nuclear antigen and DNA synthesis in a human epithelial cell line after Epstein-Barr virus infection. AB - The association of Epstein-Barr virus (EBV) with nasopharyngeal carcinoma is supported by the presence of EBV genomes in the epithelial elements of the tumor and by elevated antibody titers to EBV-specific antigens in the patients; the levels of these titers are related to the clinical course of the disease. However, since most laboratory data suggest that EBV is a B-lymphotropic virus, it is unclear how the virus becomes associated with the epithelial elements of the nasopharynx. The purpose of the present work was to find a human model system to study this association. A human epithelial line (U) was found that could be directly infected by EBV, and viral functions, the induction of EBV nuclear antigen and cellular DNA synthesis, were demonstrated. The U line was established in 1957 by the late H. J. Van Kooten (Kok-Doorschodt at the University of Utrecht), and although it is no longer diploid, it exhibits density inhibition. When U cells were infected with EBV, EBV nuclear antigen was expressed in 6 to 16% of the cells, 1 and 2 days after infection with B95-8 virus, but not with the P3HR-1 strain. No evidence for virus replication was obtained; immunofluorescence staining for early antigens and virus capsid antigens gave negative results. Quantitative adsorption experiments for EBV indicated that the adsorption capacity of U cells is significant (60% of Raji cells). The present results also demonstrated that infection with the virus overcomes block(s) in cellular DNA synthesis caused by 5-fluorodeoxyuridine. The induction of DNA synthesis was determined by increased incorporation of [3H]thymidine into the cells. The highest level of isotope incorporation was observed at about 15 h after infection and thereafter decreased. Analysis of the induced DNA indicated that it was of cellular origin. PMID- 6281480 TI - Uniform nomenclature for monoclonal antibodies directed against virus-coded proteins of simian virus 40 and polyoma virus. AB - A uniform nomenclature has been agreed upon for monoclonal antibodies directed against virus-coded proteins of simian virus 40 and polyoma virus. Blocks of numbers from PAb1 to PAb999 have been allocated to workers involved in the isolation of monoclonal antibodies of this type. The correspondence between PAb numbers and previously used names is given. PMID- 6281481 TI - RNA of rotavirus: comparison of RNAs of human and animal rotaviruses. AB - Single-stranded RNA (ssRNA) was transcribed in vitro from inner-shell particles of human rotavirus strain Wa (HRV-Wa) and a bovine rotavirus (neonatal calf diarrhea virus [NCDV]) by virion-associated RNA polymerase activity. The ssRNA product consisted of 11 RNA segments which were separated by polyacrylamide gel electrophoresis. In vitro-transcribed 32P-labeled ssRNA was used to study the genetic relatedness between rotaviruses by annealing with genomic double-stranded RNA (dsRNA) of homologous or heterologous rotavirus. All segments of HRV-Wa ssRNA were hybridized with dsRNA of HRV TK80, collected from the feces of a gastroenteritis patient, at the level of 88 to 100% of the homologous reaction. On the other hand, no segments of ssRNA from HRV-Wa hybridized with dsRNA of NCDV or simian rotavirus (simian agent 11). Similarly, ssRNA from NCDV did not hybridize with dsRNA of HRV-Wa, but hybridized with dsRNA of simian agent 11 at the level of 30% of the homologous value. PMID- 6281482 TI - Primate origin of the cell-derived sequences of simian sarcoma virus. AB - We sought to identify the species of origin of the cell-derived (sis) sequences of simian sarcoma virus. A molecular clone comprised of sis DNA detected related nucleotide sequences at low copy numbers in normal cellular DNAs of species as diverse as humans and quail. The extent of hybridization and degree of base-pair matching with sis DNA were greatest with New World primate DNAs. The thermal denaturation curve midpoints of hybrids formed between sis and woolly monkey DNAs were indistinguishable from homologous sis DNA hybrids, establishing the woolly monkey (Lagothrix spp.) as the source of sis sequences. In comparative studies, sis was shown to be more conserved among mammalian species than unique-sequence woolly monkey cellular DNA. There was no detectable homology between sis and the cell-derived sequences of other fibroblast-transforming retroviruses. These findings indicate that sis is likely to be a unique onc gene among transforming retroviruses. PMID- 6281483 TI - Identification of the polyprotein termination site on encephalomyocarditis viral RNA. AB - We show by sequence analysis of a 420-base-long region adjacent to the 3' polyadenylic acid of encephalomyocarditis viral RNA and by carboxy terminus analysis of protein E that the termination site of encephalomyocarditis virus polyprotein translation consists of two successive UAG codons located at positions 121 to 126 from the 3' polyadenylic acid. PMID- 6281484 TI - THC: "transitional drug' for emesis therapy? PMID- 6281485 TI - How will patients use acyclovir to treat herpes? PMID- 6281486 TI - Regulation of aldosterone secretion by a new aldosterone stimulating factor. AB - The site of action in the steroidogenic pathway of aldosterone stimulating factor (ASF), isolated from human urine, was studied in collagenase-dispersed rabbit adrenal capsular cells and compared with those of beta-lipotropin (beta-LPH), angiotensin II (A II) and adrenocorticotropic hormone (ACTH). When incubated with adrenal cell suspension at 37 degrees C for 2 hours, ASF, beta-LPH and ACTH induced dose-related increases in aldosterone production. ASF was less potent (ED50 = 10(-9) M) than ACTH but was more potent than beta-LPH. When ASF was added to the incubation with low dose of A II or ACTH, its effect on aldosterone production was additive, while no additional effect of ASF on aldosterone production was obtained in the presence of high dose of A II or ACTH. ASF increased the conversion of corticosterone to aldosterone like ACTH and beta-LPH. We have reported that ASF is a true aldosterone secretagogue and readily distinguishable from ACTH, A II and beta-LPH. The present study suggests ASF shares a common rate-limiting final pathway of steroidogenesis, which may be the step between corticosterone to aldosterone, with ACTH, A II and beta-LPH. PMID- 6281487 TI - [Concentration of 99mTc methylen-diphosphonate in pulmonary small cell anaplastic carcinoma and hepatic metastasis from pulmonary adenocarcinoma. Report of two cases (author's transl)]. PMID- 6281488 TI - [Wilms' tumor in an adult (author's transl)]. PMID- 6281489 TI - [Evaluation of radiological procedures in pancreatic islet cell tumor (author's transl)]. PMID- 6281490 TI - [Diagnostic value of arteriography of the primary hepatic tumor in infants and children (author's transl)]. PMID- 6281491 TI - [Outgrowth culture of carcinoma in situ (author's transl)]. PMID- 6281492 TI - [Clinicopathological and electron microscopical studies on extramammary Paget's disease. Report 1. Clinicopathological and histochemical studies of extramammary Paget's disease (author's transl)]. PMID- 6281493 TI - [Clinicopathological electron microscopical studies on extramammary Paget's disease. Report 2. Ultrastructure of extramammary Paget's disease (author's transl)]. PMID- 6281494 TI - [Clinicopathological and electron microscopical studies on extramammary Paget's disease. Report 3. Comparative studies among genital, mammary and perianal Paget's diseases (author's transl)]. PMID- 6281495 TI - [Serum sialic acid levels in liver cirrhosis and liver cancer (author's transl)]. PMID- 6281497 TI - [Detection of arterio-portal shunt in hepatocellular carcinoma by CT angiography (author's transl)]. PMID- 6281496 TI - [Real-time ultrasonographic diagnosis of hepatocellular carcinoma. Correlation of echograms and histopathological findings (author's transl)]. PMID- 6281498 TI - [Reproducibilities of the successive first pass radionuclide angiography using pertechnetate-99m (author's transl)]. PMID- 6281499 TI - [Validity of left ventricular edge-detection methods in ventricular phantom and clinical cases (author's transl)]. PMID- 6281500 TI - Effects of sodium depletion on contractions evoked in intact and skinned muscles of the guinea-pig mesenteric artery. AB - In the guinea-pig mesenteric artery, reduction in [Na]o by 30 mM (substituted by choline or sucrose; 137 mM [Na]o in Krebs solution) generated contraction with no change in membrane potential. In NaCl-free solution (15 mM [Na]o), the amplitude of phasic contraction reached 0.8 times the contraction evoked by 118 mM [K]o with only a slight depolarization. In NaCl-free solution, the amplitude of phasic contraction evoked by noradrenaline (NA) 5 X 10(-5) M or caffeine 5 mM increased to roughly twice the amplitude of the contraction evoked in the control solution. In Ca-free solution, the K-, NaCl-free- or Na-free-induced contractions rapidly ceased, but NA-induced contraction ceased within 5 min and the caffeine-induced contraction persisted for more than 15 min. In a skinned fiber, increase of [Na]o from 10 to 60 mM suppressed the pCa-tension relationship in the ranges of 10(-7) and 10(-5) M free Ca but not with a dose of 30 mM [Na]o. NA (10(-5) M) had no effect on skinned fibers. Increase in Na concentration (60 mM) had no effect on Ca accumulation in the store site or on Ca release by caffeine. Possible Na related mechanisms on the development of mechanical response are discussed in relation to Ca on the surface and in the internal membrane structure. The NaCl free-induced contraction in smooth muscles of the guinea-pig mesenteric artery is postulated to be due to influx of Ca through the Na channel, rather than the Ca channel. PMID- 6281501 TI - Role of ions in generation of taste nerve responses to electrical tongue stimulation in rats. AB - The role of ions in the generation of taste nerve responses to electric currents was studied by examining integrated responses and single fiber discharges of the chorda tympani nerve to chemical and electrical stimulations of the tongue in rats. Integrated chorda tympani responses to anodal currents were suppressed after cessation of the blood flow to the tongue and on application of FeCl3 to the tongue surface. These findings indicate that electric currents do not stimulate the nerve directly but activate taste cells to evoke neural discharges. Magnitude of the integrated response to anodal current decreased with a decrease in concentration and with an increase in flow rate of the bathing solution applied over the tongue during electrical stimulation. All the chorda tympani fibers responsive to a certain electrolyte responded to anodal or cathodal current in the presence of the same electrolyte at its subliminal concentration in the flow chamber. However, there were a few fibers which did not respond to a few electrolytes, but responded to currents in the presence of either one of the electrolytes even at their much lower concentrations. Across-fiber correlations between the responses to chemical stimuli and anodal currents in the presence of the chemical solutions in the flow chamber indicated that responsiveness to anodal current was not always dependent on species of cation in the bathing solution. From these findings, we assume that the response to electric current is due to the effect of electrophoretically carried ions on not only specific but also non-specific receptor mechanisms for the ions. PMID- 6281502 TI - Interrelation between membrane transport and the contents of alkali metal cations in HeLa cells. AB - The relation of membrane transport of alkali cations to their external concentrations or to their cellular contents was studied in HeLa cells. Chilling the cells at 0 degrees C reversed cell Na+ and K+ to a mirror image of the normal pattern. Upon rewarming to 37 degrees C the ouabain-sensitive Rb+ uptake became 2-fold faster than the control. A kinetic analysis revealed that the stimulation was due to an increase in the maximal rate of Rb+ uptake, Jmax. The increase in apparent Km was relatively small. The analysis also showed that the ouabain sensitive cation transport system seemed to have two binding sites for Rb+. The stimulation of Rb+ uptake was related to an increase in cell Na+, and an addition of ouabain abolished such a relation. Net Na+ flux which was in the direction from inside the cells to the medium at hypernormal cell Na+ was iiincreased when cell Na+ ncreased. In contrast, net Na+ flux which was in the opposite direction in the presence of ovabain was reduced and became almost 0 at cell Na+ of 900 nmol/mg of protein. The Na+/Rb+ coupling ratio in the ouabain-sensitive cation transport was apparently less than 1 at nearly physiological cell Na+, but it approached 1.5 when cell Na+ was sufficiently high. The sum of cell K+ plus Rb+ varied inversely with cell Na+, and this relation was unaffected upon treatment with ouabain. When Rb+ uptake declined below 80% of the control, cell K+ plus Rb+ was reduced, however, 40% of the sum of cell cations was still preserved even after complete inhibition of the cation pumps by ouabain treatment of 2 hr. Interrelations of these results are discussed. PMID- 6281503 TI - Effect of cholecystokinin on thyrotropin releasing hormone (TRH)-induced serotonin potentiation in rats. AB - Cholecystokinin octapeptide (CCK-8) did not affect the behavioral syndrome of rats treated with pargyline and 5-hydroxytryptamine, while TRH produced a marked behavioral excitation under the same condition. However, the serotonin potentiating effect of TRH was apparently suppressed by simultaneous administration of CCK-8. PMID- 6281504 TI - Effects of ouabain, lithium, and cooling on the frog lens fiber potential. AB - The effects of ouabain, Li+, and cooling on the lens fiber potentials i the anterior and posterior sides were investigated in American bullfrog lens mounted in a special holder by using a conventional micropipette technique. Ouabain depolarized the lens fibers in the anterior and posterior sides in a dose- and time-dependent fashion. Similarly, the fibers at both sides were depolarized in Na free, Li Ringer. The posterior fibers were depolarized faster than anterior ones during exposure to Ringer solution containing ouabain and Li+. The potential difference observed in the lens fibers of the anterior and posterior sides reduced and disappeared during a successive long exposure to ouabain and Li+. It was concluded that in the presence of ouabain and Li+ the transient potential difference between the anterior and posterior lens fibers depends on the anterior epithelial cell layer which delays the penetration of ouabain and Li+ into the lens interior, but not to the difference of total lens surface area in the anterior and posterior exposed to ouabain and Li+. Thermal dependence of the lens fiber potentials at the anterior and posterior sides was 0.78 mV/ degrees C, which is greater than the physical change of 0.2 mV/ degrees C based on the Nernst equation. PMID- 6281505 TI - The voltage-dependent nature of the GABA-induced conductance change recorded from the ganglion cell of Aplysia. AB - gamma-Aminobutyric acid (GABA) produces 2 types of hyperpolarizing responses in Aplysia ganglion cells: One is fast and Cl(-)-dependent whereas the other is slow and K+-dependent. This experiment was performed only on the cells which showed the fast, Cl(-)-dependent receptor activity. GABA-induced increase in membrane conductance (delta G) was evaluated under the voltage clamp at different potential levels of the resting membrane. The delta G was found to decrease when the membrane was hyperpolarized. The more the membrane was hyperpolarized, the greater was the decrease. The dose-response curve shifted to the right when the resting membrane was hyperpolarized, a fact suggesting an increase in apparent dissociation constant (Kapparent) of the receptor-GABA complex. In fact, Kapparent increased e-fold for every 11-mV hyperpolarization of the resting membrane. In contrast, neither parameter of other receptor activities, such as the rate of desensitization, the intrinsic activity, and the Hill coefficient, was altered by hyperpolarization of the receptor membrane. It was concluded that the voltage dependent nature observed in this type of GABA receptor might be due to the change in stability of the Cl(-)-channel in an open state at the receptor membrane. PMID- 6281506 TI - Rapid pressure changes and surface displacements in the squid giant axon associated with production of action potentials. AB - By using both optical and mechano-electric detectors, we have shown that the squid giant axon swells when an action potential is generated. The maximum swelling is reached at the peak of the action potential. The undershoot of the membrane potential is associated with a marked shrinkage of the axon. We have also demonstrated these mechanical changes in axons from which a major portion of the axoplasm has been removed. We have examined the effects of changing the tonicity of the external medium and of applying several chemical reagents. PMID- 6281507 TI - Sedative action of cholecystokinin octapeptide on behavioral excitation by thyrotropin releasing hormone and methamphetamine in the rat. AB - Intracerebroventricular (i.c.v.) injection of C-terminal octapeptide of cholecystokinin (CCK-8) in rats prolonged pentobarbital- and ethanol-induced sleeping time, but non-sulfated CCD-8 (CCK-8-NS) had no effect and caerulein showed a tendency to prolong the pentobarbital narcosis. On the other hand, i.c.v. injection of thyrotropin releasing hormone (TRH) shortened the sleeping time and the effect of CCK-8 was apparently antagonized by combined administration of TRH. Spontaneous locomotor activity in the late morning and early afternoon was not affected by CCK-8, but it increased following i.c.v. injection of CCK-8-NS. Hyperactivity produced by TRH and methamphetamine was suppressed by i.c.v. injection of CCK-8, while CCK-8-NS showed a tendency to enhance the methamphetamine-induced hyperactivity and caerulein had no effect. These results indicate that CCK-8 has a sedative action and antagonizes the behavioral excitation caused by TRH and methamphetamine, but that the effects of CCK-8-NS and caerulein were rather the opposite of those of CCK-8. In an additional experiment the TRH-induced body shaking response was not affected by combined administration of CCK-8. PMID- 6281508 TI - High-risk infants: the need for nursing follow-up. AB - The goal of perinatal care can no longer focus only on the medical management of mother and infant, but instead must take a more global approach and focus on the entire family. One aspect of family care is to provide comprehensive nursing follow-up to the high-risk family at the time of the mother's and infant's discharge. To meet this need, a program was developed to educate all interested community nurses to the specific needs of the high-risk family. Evaluation indicates that the program helped nurses to provide consistent comprehensive care to these families, who were very grateful for home visits by a community nurse. PMID- 6281509 TI - Ritodrine hydrochloride in the control of premature labor. Implications for use. PMID- 6281510 TI - Anderson--Fabry disease. PMID- 6281511 TI - [Prostanoids in the physiology and pathology of the cardiovascular system]. PMID- 6281512 TI - [Peripheral blood renin-angiotensin system indices and the kallikrein-kinin system of the kidneys in hypertension]. AB - In III patients with hypertension the activity of renin in blood plasma and excretion of kallikrein and kinins in urine and in 53 the activity of carboxycatepsin were determined. Hypertensive patients showed enhanced renin activity and excretion of kallikrein and kinins with urine. A more significant rise of these figures is noted during the early stage of illness, with labile arterial hypertension. In stable hypertension renin activity is lower than in the labile form, however this index remained higher than in the control group. Activity of kallikrein-kinin system and of carboxykatepsin in stable hypertension drops below normal. PMID- 6281513 TI - The free radical signal of pigment gallstones. AB - Bile stones have been studied by Electron Spin Resonance (ESR) spectroscopy. Pigment containing stones show a 8.76 +/- 0.02 gauss wide free radical singlet at g approximately 2.003 region of the spectrum. Amplitude of this free radical signal linearly correlates with pigment content of the stone, which can be applied in the estimation of pigment content. The possible role of free radicals is suggested in the formation of pigment bile stones. PMID- 6281514 TI - Elevated angiotensin-I-converting enzyme (ACE) in patients with essential hypertension. AB - We investigated the alteration of the ACE in different parts of the circulation in 21 patients with essential hypertension, who suffered from angina pectoris attacks. Blood samples were taken during diagnostic cardiac catheterisation. The ACE was fluorimetrically measured and compared to 48 normotensive patients. In 11 patients the Plasma Renin Activity (PRA) was additionally determined by means of bioassay. The ACE was significantly (p less than 0.001) elevated in all investigated regions but a different distribution was not observed. We found a positive correlation between the ACE from the left ventricle and the systolic, mean arterial and diastolic blood pressure. Furthermore, we observed a negative correlation between ACE and PRA. No relationship could be calculated between ACE and electrolytes, creatinine or haemodynamic parameters. Our results indicate that the ACE may contribute to the pathogenesis of so-called essential hypertension. PMID- 6281515 TI - Parathyroid gland function in subgroups of metabolically mediated urolithiasis as evaluated by serum parathyroid hormone, and urinary and nephrogenous cyclic nucleotides. PMID- 6281517 TI - [The aged: the desire for and the proximity of death]. PMID- 6281516 TI - Receptor-mediated low-density lipoprotein catabolism. AB - Low-density lipoprotein (LDL) receptors are demonstrable in cultured fibroblasts from normal subjects but are decreased or absent in cells from patients with heterozygous or homozygous familial hypercholesterolaemia. In vivo receptor mediated LDL catabolism, determined as the difference between the turnover rates of 125I-LDL and 131I-LDL coupled with cyclohexanedione, is responsible for approximately one-third of the total catabolism of LDL in normal subjects, but less than one-fifth in heterozygotes and is totally absent in homozygotes. Receptor-mediated catabolism can be stimulated in normal subjects and in heterozygotes by measures that promote bile acid synthesis, namely, administration of anion-exchange resins or creating a partial ileal bypass. Studies in dogs have shown that such measures stimulate the high-affinity binding of LDL by liver cell membranes. Taken together, these observations suggest the existence of LDL receptors in human liver, the function of which is to maintain cholesterol homeostasis within the hepatocyte during periods of increased demand. Partial or complete absence of such hepatic receptors may play a major role in the pathogenesis of familial hypercholesterolaemia. PMID- 6281518 TI - [Study on school-clinical training relations. II. Information and collaboration]. PMID- 6281519 TI - [Consultation on education. Open letter to the Swiss Red Cross]. PMID- 6281520 TI - [The aged: aging in an opulent society]. PMID- 6281521 TI - [The aged in my professional experience]. PMID- 6281522 TI - [Social conditions of and assistance to the aged. I]. PMID- 6281523 TI - [Elderly patients: impressions from geriatric centers in the Zurich and Bern cantons. Progressive ideas in the home for the aged]. PMID- 6281524 TI - [Elderly patients: the nursing service in nursing homes]. PMID- 6281525 TI - [Elderly patients: the Basel research project. The participation of the family in the care of elderly, chronically ill patients]. PMID- 6281527 TI - [Elderly patients: the grumbler]. PMID- 6281526 TI - [Elderly patients: participation of family. Nurses come to grips with the Basel research project]. PMID- 6281528 TI - [Psychiatric reform: a glimpse of the endeavors in Italy and other countries]. PMID- 6281529 TI - [Psychiatric nursing: ideal and reality]. PMID- 6281530 TI - [The aged]. PMID- 6281531 TI - [The aged: a look at old age]. PMID- 6281532 TI - [The aged: economic status of pensioners and access to health care]. PMID- 6281533 TI - [The aged: formerly, at least, one took care of the aged]. PMID- 6281534 TI - [The aged: what they think...]. PMID- 6281535 TI - [The aged: the old ladies and the animals]. PMID- 6281536 TI - [The aged: at the time of the World Assembly on Aging. Reflections about geriatrics]. PMID- 6281537 TI - [The aged: motivation of nursing personnel for the care of the aged]. PMID- 6281538 TI - [The aged: routine, the number one hazard in geriatrics]. PMID- 6281539 TI - [The aged: my work is gratifying]. PMID- 6281541 TI - [The aged: the nurse's intrusion into personal space]. PMID- 6281540 TI - [The aged: time and space]. PMID- 6281542 TI - [The aged: a group of patients with dementia]. PMID- 6281543 TI - [Study of the long-term needs of the aged of the Echallens district]. PMID- 6281545 TI - [Groupement du nursing europeen. Statutory meeting, Dublin, 2 - 5 October 1981]. PMID- 6281544 TI - [Study on school-clinical training relations. III. Preparation of nurses for their teaching role]. PMID- 6281546 TI - [Social conditions of and assistance to the aged. II]. PMID- 6281547 TI - [A taboo for nurses? Death and dying]. PMID- 6281548 TI - [Pediatric nursing. Childbirth at risk]. PMID- 6281549 TI - [Home nursing - contribution in Basel: initial experiences]. PMID- 6281550 TI - [As operating room nurse in Peru]. PMID- 6281551 TI - [Housekeeping service in the hospital: your motivation for team work with other hospital services. Housekeeping employee in the hospital]. PMID- 6281552 TI - Diagnosis of sialodacryoadenitis virus infection of rats in a virulent enzootic outbreak. AB - In a natural outbreak of sialodacryoadenitis virus it was observed that the incidence of clinical signs in spontaneous-hypertensive rats was 100%, and that these signs were of a severity not observed before in other strains of rats. Rats free of the virus were introduced so that the progress of the disease could be studied under natural conditions of spontaneous spread from the enzootically affected breeding colony. The pathogenesis of the infection in these Sprague Dawley rats has been recorded over a period of 10 days after their introduction to the colony, and the results of extensive serological screening have shown that the antibody response of the spontaneous-hypertensive rats to the virus is lower than in other strains of rat. PMID- 6281553 TI - Use of mouse hepatitis virus antigen in an enzyme-linked immunosorbent assay for rat coronaviruses. PMID- 6281554 TI - Acute myelomonocytic leukemia in a Capuchin monkey (Cebus apella). PMID- 6281555 TI - Acute gastric dilatation in common marmosets (Callithrix jacchus). PMID- 6281556 TI - Mousepox (infectious ectromelia): past, present, and future. AB - Mousepox is an orthopoxvirus infection of mice that was discovered in laboratory mice in England in 1930. Depending upon mouse genotype, it may produce a severe disease with acute hepatitis and high mortality, a generalized rash in animals that survive longer, or a trivial inapparent infection. It has long been enzootic in breeding stocks of mice in Europe, Japan, and China but not in North America and Australia. However, it has been imported into the USA on several occasions, sometimes causing severe epizootics. It may contaminate or replace various viruses that are passaged in mice and may be transferred between mouse stocks in intact mice or in mouse tumors or tissues. Vaccination with vaccinia virus provides protection and has been used to eradicate virus from mouse colonies. Depopulation and sterilization of infected animal quarters my be required. PMID- 6281557 TI - Mousepox epizootic in an experimental and a barrier mouse colony at Yale University. PMID- 6281558 TI - Observations of an outbreak of mousepox in laboratory mice in 1979 at the University of Utah Medical Center, USA. PMID- 6281559 TI - Mousepox-National Institutes of Health experiences. PMID- 6281560 TI - Clinical, pathologic, and serologic features of an epizootic of mousepox in Minnesota. AB - An epizootic of mousepox in two colonies of mice was described. Clinical signs, morbidity and mortality rates, and necropsy lesions were substantially influenced by husbandry practices, intercurrent diseases (Sendai pneumonia, mouse hepatitis virus), and total body x-irradiation. In one colony, 54 mice were necropsied; 19 mice had cutaneous or visceral lesions of mousepox although none had combined visceral and cutaneous lesions. In the other colony, 20 mice were necropsied, and 10 mice had cutaneous lesions; none had visceral lesions. Sera from 24 mice with the cutaneous form of the disease had no demonstrable HI antibody titer. Experimental mice injected with spleen/liver homogenates from infected colony mice developed typical visceral lesions with numerous poxvirus profiles on electron micrographs and positive HI titers. Mice immunized with vaccinia virus were not susceptible to the disease. PMID- 6281562 TI - Mousepox at St Louis University--preliminary report. PMID- 6281561 TI - Preliminary report of an outbreak of ectromelia (mousepox) at Washington University in St Louis. PMID- 6281563 TI - Control of mousepox epizootics in St Louis and Chicago. AB - In September 1980, mousepox was diagnosed in mice from the Jewish Hospital in St Louis, Missouri. The disease was eliminated by vaccination with vaccinia virus. In October 1980, mousepox was suspected in mice located in an off-campus research facility of the University of Chicago. Based on history, clinical signs, serology, and characteristic pathological lesions, mousepox was diagnosed. The disease was eliminated by killing or moving all mice from the facility. During serological surveillance of mice from other University of Chicago campus facilities, mice from an on-campus facility were found to have positive serological titers and pathological lesions suggestive of ectromelia infection. Some mice in this facility originated from a non-commercial source in which mousepox recently was discovered. PMID- 6281564 TI - Clinical manifestations of mousepox in an experimental animal holding room. AB - A study of the clinical aspects of mousepox was conducted during the 1979-80 outbreak at the National Institutes of Health. The disease was detected serologically in a room located adjacent to the index room. The index room received animals prior to this outbreak from a noncommercial colony which later was found to be infected with mousepox. The infection was present in the room for at least 6 weeks prior to the completion of the study. The paucity of clinical signs and low mortality were striking when compared to previous descriptions of mousepox in the United States. Only 27 of the 939 mice in the room were infected, and only one of these had typical skin lesions. A few of the mice had non specific signs such as ruffled hair coat and hunched appearance. Minimal spread of the disease was evidenced by clustering of infected cages on one of five animal racks in the room. PMID- 6281565 TI - Pathology and diagnosis of mousepox. AB - The pathologic changes of mousepox were studied during an outbreak at the National Institutes of Health in 1979. The most consistent lesions were necrosis of lymphatic tissues, especially the spleen, lymph nodes, and Peyer's patches. Hepatic necrosis and jejunal hemorrhage also were found. In two transmission studies, the disease was experimentally induced in BALB/cAnN and C3H/HeN-nu mice. Athymic mice were found to be highly susceptible, and they developed fulminant disease. The diagnosis was confirmed by demonstration of pox virions in infected tissues by electron microscopy, staining of viral antigen by immunoperoxidase methods, and by isolation of the virus in chorioallantoic membranes of hen's eggs and in cultures of chick embryonic cells. PMID- 6281566 TI - Epizootiology of an outbreak of mousepox at the National Institutes of Health. PMID- 6281567 TI - Prevention and control of mousepox. AB - The prevention and control of mousepox begins with investigators being aware of the disease's existence and its serious nature for users of mice and mouse tissues. Every institution must include as part of its management policy, provisions to ascertain that mice and mouse tissues entering its facilities are free of ectromelia virus. Policy must encompass the entire institution, be clearly stated, and must be workable. The provisions of this policy include the establishment of a disease surveillance program which actively searches for disease or virus in incoming mice and mouse tissues from untested sources. Complete records of receipts of animals and animal tissues must be maintained. There must be open and honest communication among investigators concerning the presence of mousepox (and other diseases) in their mice and tumors. Decisions must be based on accurate information. The elimination of mousepox within an institution may be difficult, and a single plan applicable to all situations cannot be given. However, the basics of all control measures include the following: recognition that mousepox represents an institutional problem, not the problem of only a single or a few investigators; rapid notification of all investigators involved, including those who may have received infected mice; confirmation of the diagnosis and notification of the scientific community of the disease's existence. Strategies for dealing with mousepox depend on the nature of the mice infected. Euthanasia of mice which can be replaced and vaccination of extremely valuable breeding stock are suggested. PMID- 6281568 TI - Summary. Ectromelia (mousepox) in the United States. PMID- 6281569 TI - Mouse hepatitis virus immunofluorescence in formalin- or Bouin's-fixed tissues using trypsin digestion. AB - Mouse hepatitis viral antigens were demonstrated by immunofluorescence in formalin- and Bouin's-fixed tissues processed routinely for histopathology followed by partial digestion with trypsin. Staining was superior in tissues fixed in formalin and was not diminished in tissue sections from paraffin blocks stored at room temperature as long as 2 years. The relative ease of this procedure and the commercial availability of reagents makes this a useful technique for the definitive diagnosis of mouse hepatitis virus infection. PMID- 6281570 TI - Histiocytic sarcoma of the scrotum in a rat. AB - A 225-day-old male fourth generation rat from a developing recombinant inbred line (Lewis x Brown Norway) had a bilaterally symmetrical enlargement of the scrotum. Palpation indicated the presence of a firm lobulated mass extending from the tip of the scrotum to the abdominal wall. Bilateral nodular masses totally occupied the scrotal sacs, surrounded the testicles, and extended along the spermatic cords into the abdominal cavity. Tumor nodules also were present in the intestinal mesentery, omentum, mesenteric lymph nodes, pancreas, and lung. Histologically, the neoplasm presented a spectrum of characteristics varying from that of a granuloma with giant cells to a diffuse proliferation of spindle-shaped mononuclear cells. PMID- 6281571 TI - Poxvirus infection in a colony of common marmosets (Callithrix jacchus). AB - An epizootic poxvirus infection occurred in a colony of 80 common marmosets (Callithrix jacchus) recently introduced to a laboratory facility. Over an 18 week period, 29 of the monkeys exhibited skin lesions that persisted for 4-6 weeks. Although eight marmosets died during the outbreak, their deaths were not attributed directly to the poxvirus infection. The skin lesions developed over the entire body surface including the soles and palms. Initially characterized as erythematous papules, they quickly changed to elevated coalescing lesions with extensive scab formation. Histopathologically, the lesions revealed moderate to marked acanthosis, and they progressed to full-thickness epidermal necrosis and ulceration. Intracytoplasmic inclusion bodies were observed occasionally within degenerate keratinocytes. These inclusions most probably constituted the intracytoplasmic aggregates of viral particles observed ultrastructurally and confirmed as members of the poxvirus group by negative staining of direct skin scrapings. PMID- 6281572 TI - Protective effect of inactivated virus vaccine on Sendai virus infection in rats. AB - Rats which had received two doses of inactivated Sendai virus vaccine were resistant to intranasal virus challenge. Vaccination was equally effective when administered by intravenous, intraperitoneal, intramuscular, or subcutaneous routes but not the intranasal route. Rats vaccinated intraperitoneally with inactivated Sendai virus vaccine were protected from contact infection. Suckling rats born to vaccinated dams were resistant to challenge infection at 3 weeks of age, but the resistance was not demonstrated after weaning at 4 weeks of age. PMID- 6281574 TI - Spontaneous hepatocellular neoplasms and hepatic hemangiosarcomas in several strains of mice. AB - The incidence of spontaneous liver tumors, ratio of hepatocellular adenomas to carcinomas, degree of differentiation of hepatocellular carcinomas, and incidence of pulmonary metastases in several strains of mice from different experiments were examined. Hepatocellular tumors were rare in all strains before 12 months of age and then increased with age. They were more common in male than in female mice. Pulmonary metastases were rare before 24 months of age and occurred in 10 of 221 mice with hepatocellular carcinomas (4.5%); their incidence in different strains was not strikingly different. Hepatic vascular tumors were less common than hepatocellular tumors. No tumors of bile duct origin were observed in these mice. PMID- 6281573 TI - A comparison of neutralization tests for the detection of antibodies to Herpesvirus simiae (monkey B virus). AB - Neutralization tests used in one laboratory in the USA and one laboratory in England to detect antibodies to Herpesvirus simiae have been compared. Complete concordance in results was obtained with 53 (90%) of 59 monkey sera. The remaining six sera all had titers no greater than 1:3. Four were positive only in the American test, and two were positive only in the British test. The importance of using complement if maximum sensitivity is to be achieved in detecting antibodies to this virus has been confirmed. PMID- 6281575 TI - Nephroblastoma with associated aortic rupture in a rat. AB - A nephroblastoma was diagnosed in a 6-month-old, male rat which died unexpectedly. The right kidney was replaced by a large, white, irregularly shaped mass. Microscopically, the mass consisted of an embryonic blastema showing epithelial differentiation into immature tubules and glomeruloid structures. An aortic aneurysm and aortic rupture were present within the thoracic cavity. It was suspected that the aortic rupture was secondary to hypertension mediated through the renin-angiotensin system. PMID- 6281576 TI - Confirmation of a tetrahydrocannabinol metabolite in urine by gas chromatography. AB - A new method for confirming urinary 11-nor-delta 9-tetrahydrocannabinol-9 carboxylic acid, the major metabolite of tetrahydrocannabinol, has been developed. The metabolite is extracted, derivatized to the methyl ester, methyl ether, and analyzed on a gas chromatograph equipped with a flame ionization detector. In spiked urine specimens, metabolite concentrations as low as 20 ng/mL have been detected by this procedure. In a random sampling of urines, greater than 95% correlation was obtained between confirmation by this method and confirmation by gas chromatography/mass spectrometry in those specimens producing an immunological response greater than 75 ng/mL. PMID- 6281577 TI - Interaction of fluid movement and particle diffusion across capillary walls. AB - The rate of transcapillary exchange of substances delivered to or removed from the tissue by blood depends on the concentration difference across the capillary wall. This concentration difference results in an osmotic effect that can have considerable influence on capillary-tissue fluid exchange. However, fluid exchange also affects the plasma and tissue concentrations of the various substances. Therefore, the determination of substrate concentration profiles involves an interaction of many microcirculatory phenomena, including capillary tissue fluid exchange. In this paper a general mathematical model is presented that described the interaction of multiple exchangeable solutes. The resulting equations are solved to give the concentration profiles of different substances and their effect on fluid exchange for various normal and pathological situations of physiological interest. PMID- 6281578 TI - Studies on high affinity binding of mineralo- and glucocorticoids in rabbit aorta cytosol. AB - High affinity, specific binding-sites to mineralocorticoids and glucocorticoids, with characteristics of steroid receptors, have been found in rabbit aorta cytosol. Binding parameters (dissociation constants and number of binding sites per mg of cytosol protein) were determined from Scatchard plots, after statistical treatment of the data with the aid of a computer program, for the following tritiated steroids: 11-desoxycorticosterone (DOC), aldosterone (Aldo), progesterone (Prog), corticosterone (BK), cortisol (FK) and dexamethasone (Dex). The specificity of binding was then examined by means of steroid competition studies. The results of these experiments indicate that three different types of high-affinity binding sites to adrenal steroids are present in aorta cytosol: Type A, with the highest affinity for DOC; Type B, with the highest affinity for FK; Type C, with the highest affinity for Dex. In accordance with the relative competitive potencies of various steroids for these binding sites, Type A is designated as the "arterial mineralocorticoid binder", clearly differing in its binding characteristics from the cytoplasmic mineralocorticoid binders in known target tissues to these steroids (e.g. the renal receptor), while Type C is designated as the "arterial glucocorticoid binder", closely resembling the classical glucocorticoid receptor in known target tissues to glucocorticoids. Type B exhibited some of the binding characteristics of transcortin and may represent a modified, intracellular transcortin. While Types B and C are present also in the cytosol of inferior vena cava. Type A was only in the aorta. The role of these binders is not known at present. Arguments are presented in favor of a hypothesis that the Type A (mineralocorticoid) binder represents an arterial wall; and that, under certain conditions, this action leads to an increased contractility of arterial and arteriolar smooth muscles, increased peripheral resistance and hypertension. PMID- 6281579 TI - Biosynthesis, purification and receptor binding properties of high specific radioactivity 1 alpha, 24(R),25-trihydroxy-[26,27-methyl-3H]-vitamin D3. AB - A procedure for the biosynthesis and purification of 1 alpha, 24(R),25 trihydroxy[26,27-methyl-3H]-vitamin D3 (1,24,25-(OH)3[3H]D3) is reported. A kidney homogenate from chicks receiving a high calcium diet (3%) and oral supplements of 1 alpha, 25-dihydroxyvitamin D3 (1,25-(OH)2D3) was used for C-24 hydroxylation of 1 alpha, 25-dihydroxy[26,27-methyl-3H]-vitamin D3 (1,25 (OH)2[3H]D3), in vitro. Extraction and purification of the homogenate lipid fraction by Sephadex LH-20 and high performance liquid chromatography yielded radiochemically pure 1,24-25-(OH)3[3H]D3 with a specific radioactivity equivalent to the initial substrate (166 Ci/mmol). The authenticity of the generated metabolite was assessed by co-migration with synthetic 1,24,25-(OH)3D3 on high performance liquid chromatography and by equimolar competition with authentic radioinert 1,24,25-(OH)3D3 for binding to a purified receptor protein from rat kidney. Binding studies indicate the trihydroxylated metabolite competes 40-50% as effectively as 1,25-(OH)2D3 for hormone binding sites. Further analysis of 1,24,25-(OH)3D3-receptor interaction reveals a high-affinity, saturable binding with an apparent K4 of 2.2 x 10(-9) M. These studies demonstrate that although slightly less active than 1,25-(OH)2D3, 1,24,25-(OH)3D3 is capable of hormone like interactions, in vitro. The availability of this high specific radioactivity sterol should allow for clarification of its potential physiologic significance. PMID- 6281580 TI - Inhibition of dibutyryl cyclic AMP induced steroidogenesis in rat adrenocortical cells by the putative calcium antagonist TMB-8. AB - A significant proportion of the steroidogenic response of isolated rat adrenocortical cells to dibutyryl cyclic AMP does not require extracellular calcium, and this component is profoundly depressed by low concentrations of the putative calcium antagonist, TMB-8. The inhibition is reversed by either the readdition of calcium or the calcium ionophore A23187. The steroidogenic response to pregnenolone, whose mode of action does not require calcium, was not depressed by TMB-8. Corticotropin (ACTH)-induced steroidogenesis, which requires extracellular calcium, was markedly depressed by TMB-8, although enhanced cyclic AMP formation is only slightly depressed by this drug. Adrenal cortical microsomes possess an ATP-dependent 45calcium (45Ca2+) uptake system which responded to EGTA with a rapid efflux of 45Ca2+; EGTA-induced calcium efflux from this microsomal fraction was markedly reduced by a concentration of TMB-8 that blocked dibutyryl cyclic AMP-evoked steroidogenesis. TMB-8 produced a smaller but significant reduction of EGTA-facilitated 45Ca2+ efflux from a mitochondrial enriched fraction. We interpret these results to mean that TMB-8 blocks the steroidogenic effect of dibutyryl cyclic AMP by interfering with the mobilization of a cellular pool of calcium that is probably localized to the endoplasmic reticulum. The physiological implications of these findings in relation to the complex interactions between calcium and cyclic AMP in adrenal steroidogenesis are discussed. PMID- 6281581 TI - Esophageal granular cell tumors. Report of two multicentric cases with observations on their natural histories. PMID- 6281582 TI - Simulation of intracellular Ca2+ oscillation in a sympathetic neurone. PMID- 6281583 TI - Markov process characterization of a single membrane gating site. PMID- 6281584 TI - Macroscopic law for time and activity in a neural system. PMID- 6281585 TI - A stereochemical concept for the catalytic mechanism of prolylhydroxylase: applicability to classification and design of inhibitors. PMID- 6281586 TI - Angiofollicular lymph node hyperplasia and peripheral neuropathy: association with monoclonal gammopathy. AB - An elderly man was found to have the plasma cell variant of angiofollicular lymph node hyperplasia. His course was complicated by peripheral neuropathy and generalized lymphadenopathy, which improved after a trial of corticosteroid therapy. Although the plasma cell variant has been associated with multiple systemic effects, including the nephrotic syndrome, growth failure, fever, hyperglobulinemia, and anti-erythropoietin-mediated anemia, concurrent peripheral neuropathy has only occasionally been reported. Angiofollicular lymph node hyperplasia should be included in the differential diagnosis of peripheral neuropathy associated with lymphadenopathy or a mediastinal mass. A discussion of the clinical, histologic, and immunopathologic characteristics of angiofollicular lymph node hyperplasia is presented. PMID- 6281587 TI - Diseases of the placenta and their effect on transport. PMID- 6281590 TI - [Rheumatoid factor in liver cirrhosis and hepatoma (author's transl)]. PMID- 6281588 TI - [Arterial obstruction and pulmonary infarction as the first manifestation of pulmonary oat cell carcinoma (author's transl)]. PMID- 6281589 TI - [Hepatic coma secondary to hepatic metastases of a lung cancer]. PMID- 6281591 TI - Viral gastroenteritis. PMID- 6281592 TI - Current management of fungal enteritis. AB - Fungal infections of the gastrointestinal tract have risen to higher levels of prevalence in the past decade. Major factors accounting for this increase are social changes, such as the increased ease and frequency of travel, which exposes the individual to environmental conditions that may result in fungal infection; increasing use of antibiotic and hormonal medications by otherwise healthy persons; and improved therapy for other diseases, such as polychemotherapy of cancer with its immunosuppressive effects. Both noninvasive and invasive fungal disease of the intestinal tract in otherwise healthy individuals can be successfully treated. The invasive fungal infections in patients with severe prior underlying disease are often first diagnosed postmortem, but improvement in serologic techniques now offers a possibility of earlier diagnosis and therapeutic intervention. PMID- 6281593 TI - The presentation of amoebiasis. PMID- 6281594 TI - Gastrointestinal infections in the compromised host. PMID- 6281595 TI - Adenoid cystic carcinoma of the submaxillary gland. AB - A review of submaxillary gland neoplasms over a 21 year period revealed 15 malignancies and 7 benign tumors. All of the malignancies occurred in females, and 11 of these were adenoid cystic carcinoma. Two of the 11 were found to have infiltrating ductal carcinoma of the breast (1 pre and 1 postdiagnosis of the submaxillary carcinoma) and 3 had benign breast disease. While previous reports have suggested an association of parotid gland neoplasia and breast cancer, this is the first known report of an association between adenoid cystic carcinoma of the submaxillary gland and cancer of the breast. The discussion of adenoid cystic carcinoma of the submaxillary gland emphasizes the increased frequency of this disease in females, its association with breast disease, and also experimental submaxillary gland neoplasia. PMID- 6281596 TI - Arguments against a mediating role of the adenylate cyclase--cyclic AMP system in the ototoxic action of loop diuretics. AB - Previous studies in our laboratory have indicated that adenylate cyclase of the stria vascularis is strongly inhibited in vitro by ethacrynic acid and furosemide. In order to test whether the in vitro effects upon the enzyme are also present under in vivo conditions, ethacrynic acid was perfused perilymphatically for 15 min and 20 min at a concentration of 10(-3) M. Cyclic AMP of the stria vascularis was reduced by 27% and 34%, respectively, but ATP also declined significantly, suggesting unspecific effects. When ethacrynic acid was applied intravenously at a dosage of 50 mg/kg, and the endolymphatic potential allowed to decline to -10mV, no significant changes in cyclic AMP and ATP were seen. The absence of effects upon cyclic AMP in the early stage of systemic intoxication with ethacrynic acid is strong evidence against a mediating role of adenylate cyclase in the ototoxic action of ethacrynic acid. When a bolus of 3 x 10(-2) M furosemide was applied intra-arterially the endolymphatic potential declined at the exceedingly rapid rate of about 10 mV/sec, strongly suggesting that the action of the drug takes place in the vicinity of the capillaries of the stria vascularis. In view of the proposition that adenylate cyclase appears to be located primarily at eh luminal aspect of the stria vascularis, this constitutes further evidence against a role of the enzyme in the mediation of the specific ototoxic effects of loop diuretics. Other recent evidence against a mediatory role of the adenylate cyclase--cyclic AMP system is discussed. PMID- 6281597 TI - The use of somatic cell hybrids for the production of monospecific viral antibodies. PMID- 6281598 TI - Techniques for the cell-free translation of viral RNA. PMID- 6281599 TI - ELISA techniques in virology. PMID- 6281600 TI - Radioimmunoassay in diagnostic virology. PMID- 6281601 TI - Effect of maleate on membrane physical state of brush border and basolateral membranes of the dog kidney. AB - The effects of maleate on the physical state of isolated brush border and basolateral membranes from dog kidney cortex have been studied by fluorescence polarization and ESR methods. Anisotropy of 1,6-diphenyl-1.3.5.-hexatriene and hyperfine splitting (2T parallel) of 5-doxylstearic acid in brush border and basolateral membranes were not significantly modified by the addition of 10(-2) M maleate after 60-90 min treatment. These findings further support the view that maleic acid nephropathy is not due to a direct effect of maleate on tubular membranes per se. PMID- 6281602 TI - Modulation of the hippocampal alpha-adrenoceptor population by lesion of the serotonergic raphe-hippocampal pathway in rats. AB - Electrolytic lesion of the ascending serotonergic fibers in the median raphe nucleus or in both the median raphe nucleus and dorsal raphe nucleus caused after 18 days more than 80% depletion of serotonin in the hippocampus and frontal cortex, respectively, without affecting norepinephrine and acetylcholine contents. alpha 1-Adrenoceptor binding of (3H) WB-4104 was increased in the hippocampus but not in the frontal cortex. Scatchard analysis revealed that the increase in (3H) WB-4101 binding in the lesioned hippocampus was the result of an elevated density of alpha 1-adrenergic receptors of about 65%. This phenomenon began 8 days postlesion and persisted for at least 90 days postlesion. Similar qualitative and quantitative results were obtained following chemical lesion of the serotonergic cells of origin in the median raphe nucleus with 5,7 dihydroxytryptamine. Selectivity of the phenomenon was further demonstrated as or beta-adrenoceptor binding with (3H) dihydroalprenolol and cholinergic muscarinic receptor binding with (3H) dexetimide were not significantly affected in the hippocampus. By comparison, when norepinephrine in the hippocampus was depleted by more than 90% by bilateral lesion of the ascending noradrenergic fibers with 6 hydroxydopamine (18 days), the alpha 1-adrenoceptor number was significantly increased by only about 20% while the beta-adrenoceptor number was enhance by 40%. The area-selective increase in alpha 1-adrenoceptor number in the hippocampus in the presence of unchanged norepinephrine content and in the absence of serotonin probably signifies that serotonin actively participates in the modulation of the noradrenergic receptor population. PMID- 6281603 TI - Hypercalcemia in association with a Leydig cell tumor in the rat: a model for tumor-induced hypercalcemia in man. AB - The etiology of tumor-induced hypercalcemia was investigated in a transplantable Leydig cell tumor of the Fischer rat. In this model, serum calcium rose from a baseline of 10.4 +/0 0.3 mg/dl to 12.5 + 0.4 mg/dl at day 10 and 16.4 +/- 1.3 mg/dl (p less than 0.001) at day 13 post transplant. Urinary calcium also increased from 1.52 +/- 0.17 mg/d to 3.52 + 0.72 mg/d (Day 12, p less than 0.01). Serum phosphate decreased from a baseline of 7.5 +/- 0.3 mg/dl to 5.5 +/- 0.6 mg/dl at day 13 (p less than 0.05). At day 13 serum immunoreactive parathyroid hormone levels fell 76% from baseline (p less than 0.01). Calcitonin increased from 59 +/- 2 pg/ml to 88 +/- 9 pg/ml (p less than 0.02). The plasma prostaglandin E metabolite, 13,14-dihydro-15-keto-PGE2 increased from 407 +/- 103 pg/ml to 647 +/-62 pg/ml (p less than 0.05) and the active Vit D compound 1,25(OH)2D increased from 94.8 +/- 5.2 pg/ml to 162.3 +/- 11.8 pg/ml (p less than 0.01). Urinary cyclic AMP did not decrease in parallel with the parathyroid hormone level and, in fact, increased from 146 +/- 3 nmol/d to 172 +/- 27 nmol/d (NS). Administration of the cyclooxygenase inhibitor indomethacin (20 mg/Kg/d) or hydrocortisone (50 mg/Kg/d) did not prevent the development of hypercalcemia. This model is similar to many patients with humoral hypercalcemia of malignancy who demonstrate suppression of parathyroid hormone with elevated urinary cyclic AMP excretion and may prove useful in the understanding of the responsible mechanisms. PMID- 6281604 TI - Analgesic activity of intracerebroventricular administration of morphiceptin and beta-casomorphins: correlation with the morphine (micro) receptor binding affinity. AB - Analgesic activities of morphiceptin, beta-casomorphins, [D-Ala2, D Leu5]enkephalin and Sandoz peptide, FK 33-824, were examined by intracerebroventricular administration in rats. Their relative potencies in vivo were compared with their receptors binding activities. The receptors binding affinities were determined from the competition curves against [3H]naloxone binding in the absence and presence of sodium ions for morphine (micro) receptors and against 125I-[D-A1A2, D-Leu]enkephalin binding for enkephalin (delta) receptors. A good correlation between analgesic activity and morphine (micro) receptor but not enkephalin (delta) receptor binding affinity was obtained. These data extend the hypothesis that morphine (micro) receptors mediate the major portion of the analgesic activity of opioids. PMID- 6281605 TI - Hyperactivity versus explosive motor behavior induced by opioids in the rat. Mechanisms elucidated with enkephalin-tyrosine-o-sulfate and morphine congeners. AB - A relatively mild hyperactive state (HAS), characterized by agitation and hypermotility, is induced by opiate drugs and opioid peptides in general and is blocked by naloxone. HAS can be distinguished from the profound hyperresponsiveness of an explosive motor behavior (EMB). Sulfation of the phenolic moiety in morphine or in methionine enkephalin essentially abolishes opiate receptor binding activity. The sulfated peptide lacks detectable pharmacological activity in the rat, whereas sulfated morphine is several hundred fold more potent than morphine in eliciting (EMB). Thus, EMB is elicited only by congeners of morphine having appropriate hydrophilic substitution at C-6 and which is mediated through a receptor that is insensitive to naloxone. PMID- 6281606 TI - [Quantitative evaluation of the effectiveness of radiomodifiers in local irradiation of the skin]. PMID- 6281607 TI - [One hundred and one cases of pulmonary aspergillosis in Ivory Coast (author's transl)]. AB - Report on 101 cases of intracavitary pulmonary aspergillosis observed in Abidjan with special regard to the circumstances of the diagnosis, the radiologic aspects, and the serologic findings. This series is the longest published in West Africa. Tuberculous sequellae, often of large extension, are detected in 87 p. 100 of the cases. The symptomes of Deve's triade are reminded, and some therapeutic prospects are briefly indicated. PMID- 6281608 TI - Lipoatrophic diabetes: endocrine dysfunction and the response to control hypertriglyceridemia. AB - Endocrine function was studied in a 24 year old female with lipoatrophic diabetes (LD). Baseline endocrine studies (serum triglycerides: 2600 mg/dl) demonstrated hyperprolactinemia (serum prolactin 51 ng/ml), increased ACTH levels, absence of suppression of ACTH to a high dose of dexamethasone which suppressed serum cortisol normally and, hyperresponsiveness of TSH to stimulation with TRH. Thyroid hormone levels (total and free fraction) were essentially normal. Major metabolites of thyroid hormone (T3, rT3, 3, 3'-T2, and 3', 5'-T2) were also normal and exhibited a normal response to the administration of L-thyroxine and propylthiouracil. Exchange of 84% of the patient's plasma resulted in a decrease in serum triglycerides (700 mg/dl) which was followed by a rebound to the original level in seven days. After the sixth plasmapheresis serum triglycerides stabilized at less than 1000 mg/dl. Plasmapheresis was associated with the appearance of amenorrhea and galactorrhea; also hypertension and proliferative retinopathy developed during this therapy. Repeat endocrine function studies (serum triglycerides: 700 mg/dl) showed a further rise in serum prolactin (greater than 160 ng/ml), persistence of abnormal ACTH secretion and normalization of TSH responsiveness. Lipoatrophic diabetes is associated with abnormal central endocrine function but appropriate peripheral target gland secretion. A course of plasmapheresis improves the hypertriglyceridemia but not the endocrine dysfunction. In this patient with LD the most important side effect of plasmapheresis was the development of cardiovascular complications. PMID- 6281609 TI - Interrelationships between vitamin D3 and 25-hydroxyvitamin D3 during chronic ethanol administration in the rat. AB - Vitamin D [D] depleted female Sprague-Dawley rats were fed for a period of 4 wk a D deficient diet containing 36% of total calories as ethanol while control animals received an isocaloric regimen where ethanol was substituted for by dextrins. In conjunction with the ethanol feeding 92 I.U. of [14C]-vitamin D3 [(14C)-D3] were administered by intragastric gavage 3 times 1 wk for 3 2/3 wk. At the end of the experiment, [14C]-D3 and [14C]-25-hydroxyvitamin D3 [(14C) 25(OH)D3] concentrations were analyzed in plasma, liver, striated muscle and adipose tissue. Body reserves in unchanged [14C]-D3 were significantly reduced by ethanol treatment as seen by 24%, 26%, and 59% lower plasma (p less than 0.02), muscle (p less than 0.001) and adipose tissue (p less than 0.001) [14C]-D3 concentrations in ethanol-treated compared to control rats. In contrast total plasma and liver [14C]-25(OH)D3 content were increased by 30% (p less than 0.05) and 55% (p less than 0.001) respectively. This increased liver and plasma [14C] 25(OH)D3 following ethanol treatment was not accompanied by a proportional [14C] 25(OH)D3 incorporation into muscle and adipose tissue. These results suggest that during steady state conditions 25(OH)D3 production is increased during chronic ethanol administration while the body pool in unchanged D3 is significantly lowered. These results also point out that in the rat plasma 25(OH)D concentrations are not a reliable guide for the determination of vitamin D status during chronic ethanol administration. PMID- 6281610 TI - Dissociation of the in vitro transfers of esterified cholesterol and triglyceride between human lipoproteins. AB - This report describes the detailed time-course of in vitro net mass transfers of esterified cholesterol (EC) and triglyceride (TG) between human plasma lipoproteins incubated in the presence of EC and TG transfer proteins. In incubations containing high density lipoproteins (HDL), very low density lipoproteins (VLDL) and the thiol-group blocker parachloromercuriphenyl sulfonate (PCMPS) (which was added to inhibit EC production), the rate of EC transfer to VLDL was considerably more rapid than that of TG to HDL and equilibrium between the pools of EC was achieved much sooner than was the case for the pools of TG. When VLDL and low density lipoproteins (LDL) were incubated, the presence of PCMPS in the incubation mixture resulted in a marked reduction in the TG transfer to LDL, even though the EC transfer to VLDL was apparently unaffected. The molar ratio of the transfers of EC and TG was also examined in incubations of lipoproteins from a variety of human subjects. In incubations of HDL and VLDL (performed in the absence of PCMPS) there was a large individual variation (sixfold difference) in the molar ratio of the transfers of TG:EC. In incubations of LDL and VLDL from different subjects there was a twofold individual variation in the molar ratio of the transfers of TG:EC. It has been concluded that the net mass transfers of EC and TG between human lipoproteins do not involve a simple molecular exchange of one moiety for the other, as has been suggested previously, but are achieved by processes which are at least partially independent. PMID- 6281611 TI - Relationship between calcium stimulation of cyclic GMP and lipid peroxidation in the rat kidney: evidence for involvement of calmodulin and separate pathways of peroxidation in cortex versus inner medulla. PMID- 6281612 TI - Adverse effects of fructose in perfused livers of diabetic rats. AB - Livers isolated from both fed normal and alloxan diabetic rats were perfused for 30 min using Krebs-Henseleit bicarbonate blood buffer medium followed by 10 min flow-through infusions with either 5 mM or 28 mM fructose concentrations. In livers of normal and diabetic rats, both 5 mM and 28 mM fructose concentrations produced an elevation in tissue cyclic AMP levels, activation of glycogen phosphorylase, increased protein kinase activity, decreased tissue ATP levels, large increases in tissue fructose-1-phosphate, and variable effects upon glycogen synthase. These results are consistent with previously reported cyclic AMP mediated activation of glycogen phosphorylase by fructose via protein kinase in normal rat liver. In addition, both 5 mM and 28 mM fructose infusion resulted in large decreases in normal and diabetic synthase phosphatase activity. Therefore, these results in both normal and diabetic livers are inconsistent with a direct beneficial effect of fructose in the isolated perfused rat liver. PMID- 6281613 TI - Actin in Triton-insoluble cytoskeleton of thyroid. AB - Treatment of bovine thyroid with the non-ionic detergent Triton X-100 extracts most of the cell protein and leaves insoluble residue. This Triton-insoluble cytoskeleton consists of five major polypeptides on sodium dodecyl sulfate polyacrylamide gels. One of these polypeptides is actin. Based on DNase inhibition assay, 30% of the total actin is associated with the cytoskeleton as the filamentous form. Thyroid actin from the cytoskeleton has been solubilized by dialysis against a low ionic strength buffer at pH 8.0 and purified to homogeneity by a polymerizing-depolymerizing cycle. The overall purification was about 144-fold with a yield of 10%. Bovine thyroid actin is very similar to actins from other tissues on the basis of: (1) comigration with rabbit skeletal muscle actin during gel electrophoresis in sodium dodecyl sulfate, (2) its amino acid composition, which includes about 1 mole of 3-methylhistidine per 42,000 g, (3) its ability to bind and inhibit pancreatic deoxyribonuclease I, and (4) its ability to form 7-8 nm microfilaments which is similar to that of skeletal filamentous actin. Thyroid actin contains beta- and gamma-isoactins, with isoelectric points more alkaline than the alpha-actin of rabbit skeletal muscle. PMID- 6281614 TI - Sodium potassium dependent ATPase in hypophysectomized rats: response to growth hormone, triiodothyronine, and cortisone. AB - Groups of hypophysectomized rats were treated with pharmacologic doses of growth hormone, triiodothyronine or cortisone acetate alone or with a combination of growth hormone plus triiodothyronine or growth hormone plus cortisone. After a 7 day period of treatment the hydrolysis of ATP in the presence of ouabain (mg ATPase) and in the absence of ouabain (total ATPase) was determined. Ouabain suppressible sodium, potassium-dependent ATPase or (Na+ + K+) ATPase was calculated as the difference in the rate of hydrolysis in the presence and absence of ouabain. The activity of the Mg ATPase was significantly reduced in brain after treatment with growth hormone regardless of whether other hormone. In liver there was a significant increase in (Na+ + K+) ATPase in growth hormone, triiodothyronine, or (Na+ + K+) ATPase but there was no effect of triiodothyronine or cortisone and no interaction with the effect of growth hormone. In liver there was a significant increase in (Na+ + K+) ATPase in growth hormone, triiodothyronine, or cortisone-treated animals but Mg ATPase was unaffected by hormone treatment except for the group receiving both growth hormone and cortisone. In kidney homogenates both growth hormone and triiodothyronine significantly increased the activity of the (Na+ + K+) ATPase. There was no effect of cortisone. These data suggest that growth hormone and triiodothyronine may both be calorigenic through their effect on the sodium pumping mechanism in the call membrane. PMID- 6281615 TI - Preservation of normal adrenal androgen secretion in end stage renal disease. AB - A common endocrine defect in uremia is gonadal dysfunction with decreased testosterone production. Since gonadal and adrenal tissues share androgen biosynthetic pathways, we studied the stimulated adrenal androgen response in uremic patients. In contrast to the delayed or subnormal gonadal response to hCG reported by others, the adrenal response of androgens, as well as cortisol and aldosterone, to cosyntropin stimulation was unimpaired. In summary, the secretory reserve capacity of the adrenal gland for androgen, glucocorticoids and mineralocorticoids in uremia was studied with cosyntropin stimulation and found to be wall preserved. PMID- 6281616 TI - Studies on desensitization of adrenergic receptors of human adipocytes. AB - The studies described here were undertaken to determine whether or not desensitization of human adipocyte beta and alpha-2 adrenergic receptors could be demonstrated. Cells, isolated from peritoneal adipose tissue obtained from patients undergoing elective abdominal surgery, were preincubated for 3 hr in buffer alone or in the presence of isoproterenol, 10-5M. Cells in both sets of flasks were then washed and exposed to isoproterenol for 1/2 hr; cyclic AMP was then measured as an end point of beta receptor activation. Cells which had had no prior exposure to isoproterenol responded significantly greater to isoproterenol than did cells that had had prior exposure to the catecholamine, The beta receptor characteristics of cells undergoing beta desensitization were assessed using [3H] dihydroalprenolol. Compared to control cells, adipocytes exposed to isoproterenol had a reduction in Bmax while KD values were the same. Thus desensitization of beta adrenergic receptors of human adipocytes occurs and is associated with down regulation in the number of beta receptors. In comparable studies, preincubation with epinephrine 10-5M did not affect the response of cells to a subsequent exposure to this catecholamine. In alpha-2 receptor binding assays, there was a decreased number of [3H]p-aminoclonidine binding sites, but the level of [3H]yohimbine binding was not altered following the incubation with epinephrine. Thus, desensitization of alpha-2 receptors was not demonstrated. PMID- 6281617 TI - Investigations into the etiology of elevated serum T3 levels in protein malnourished rats. AB - Thyroid function studies and the peripheral metabolism of thyroid hormone were examined in rats fed a low protein diet (9% casein) for 4-8 wk. Compared to animals fed a normal protein diet ad libitum, both the low protein rats and a pair-fed control group weighed less at the end of the study. However, serum total T3 levels were significantly higher only in the protein deficient rats. The elevated serum T3 was not explainable by enhanced peripheral T4 to T3 conversion, as there was no evidence of any change in hepatic or renal 5'-deiodinase activity when homogenates were examined for conversion of T4 to T3, reverse T3 to 3,3' diiodothyronine, or 3',5'-diiodothyronine to 3'-monoiodothyronine. Neither was there an effect on hepatic T3 receptor maximal binding capacity (204 +/- 24 versus 168 +/- 15 fmol/mg DNA control) or binding affinity (2.07 +/- 0.38 versus 2.49 +/- 0.24 x 10(-10) M control). In two separate experiments the dialyzable fraction of T3 was significantly lower in the low protein group while free T3 concentrations were unchanged or reduced. In contrast, serum total and free T4 were either normal or reduced and dialyzable T4 was unaffected by protein deficiency. We conclude that while serum total T3 is elevated in rats chronically fed a low protein diet, this elevation is not due to enhanced T4 to T3 conversion. Rather, the increased T3 levels can be accounted for by a striking alteration in protein binding to T3. Moreover, the failure to demonstrate similar changes in serum total and dialyzable T4 suggests that in the rat, protein deficiency has different effects on binding to the two major thyroid hormones. Dietary induced changes in serum thyroid hormone binding must be kept in mind in nutrition studies in the rat. PMID- 6281618 TI - Tamoxifen inhibits Leydig cell steroidogenesis: in vivo and in vitro studies. AB - Using isolated interstitial cells from testes of Sprague-Dawley rats, we have shown previously that tamoxifen inhibits LH and 8-bromo-cyclic AMP stimulated testosterone synthesis in a dose-dependent manner. The inhibitory effect of tamoxifen could not be reversed with 17 beta-estradiol. The present studies indicate that tamoxifen directly inhibits testosterone response to gonadotropin stimulation both in immature and mature hypophysectomized rats. When interstitial cells were incubated with pregnenolone (5 x 10(-7) M), testosterone levels in the incubation medium were 27.0 +/- 1.9 ng/10(6) cells. Tamoxifen (10(-5) M) significantly inhibited pregnenolone-induced testosterone formation. Tamoxifen also significantly diminished adenylate cyclase activity whereas the binding of hCG to receptor was not affected. These results indicated that several steps of steroidogenesis are inhibited by tamoxifen. PMID- 6281619 TI - The effects of metabolic acidosis in vivo on insulin binding to isolated rat adipocytes. AB - Insulin resistance in vivo and impaired insulin binding to isolated adipocytes are characteristic of diabetic ketoacidosis in the rat. To determine the respective roles of diabetes and acidaemia in the genesis of the binding defect, insulin binding to adipocytes from alkali-treated ketoacidotic diabetic and ammonium chloride acidotic rats was studied. Reversal of the acidaemia of ketoacidotic rats by sodium bicarbonate infusion (pH 6.73 +/- 0.027 to 7.35 +/- 0.027, p less than 0.001, n = 12) increased adipocyte insulin binding (0.51 +/- 0.21% to 2 x 10(5) cells/ml, n = 6 untreated versus 1.10 +/- 0.27% to 2 x 10(5) cell, n = 6 treated, p less than 0.05). Scatchard analysis showed this to be due to an increase in insulin receptor concentration. Ammonium chloride infusion caused marked metabolic acidaemia (pH 6.72 +/- 0.04, n = 12) and insulin binding to adipocytes was markedly decreased (0.81 +/- 0.12% to 2 x 10(5) cells/ml n = 6 versus 2.40 +/- 0.22% to 2 x 10(5) cells/ml, n = 6 in controls p less than 0.02), due to a change in receptor concentration. The apparent affinity of the receptor was markedly decreased in diabetic animals compared with normal controls but was unchanged in ammonium chloride acidotic animals. Thus in diabetic ketoacidosis there is both decreased affinity and number of insulin receptors partially reversible by prolonged alkali infusion. Only changes in affinity appeared to be specific for the diabetic state. PMID- 6281620 TI - The human complement system serine proteases C1r and C1s and their proenzymes. PMID- 6281621 TI - Preparation and properties of human C1 inhibitor. PMID- 6281622 TI - Human peptidyl dipeptidase (converting enzyme, kininase II). PMID- 6281623 TI - Calcium-activated proteases in mammalian tissues. PMID- 6281624 TI - Mammalian collagenases. PMID- 6281625 TI - Vertebrate collagenases. PMID- 6281626 TI - Hepatocyte--collagen adhesion. PMID- 6281627 TI - Biosynthetic matrices from cells in culture. PMID- 6281628 TI - Phase-shift markers in the genus Bordetella: loss of cytochrome d-629 in phase IV variants. AB - The respiratory electron transport chain of Bordetella pertussis was examined in whole cell suspensions using difference spectra obtained at room temperature. Phase I (virulent) strains were found to possess cytochromes a-603, b-560, c-550, d-629 and cytochrome o. Cytochrome c-553, previously reported (Sutherland, 1963) was not detected and was assumed to be masked by the alpha-peaks for c-550 and b 560. Phase IV (avirulent) strains and C-mode cells (phase I strains grown in the presence of 20 mM MgSO4) were deficient in cytochrome d-629 and appeared to have higher levels of cytochromes a-603, b-560, c-550 and cytochrome o. Preliminary data indicate that B. parapertussis and B. bronchiseptica have electron transport chains similar to that of B. pertussis. PMID- 6281629 TI - [Wandering genes: fatal antibiotic resistance]. PMID- 6281630 TI - [ACTH response in the dexamethasone suppression test]. AB - Despite the great clinical importance of the low-dose dexamethasone suppression test (determination of cortisol after dexamethasone 2 mg p.o. at 11.00 p.m. in the evening before) and of the determination of ACTH in plasma, so far only little is known about the ACTH response to dexamethasone in normal subjects. In the present study cortisol and ACTH were determined by radioimmunoassay in normal subjects at 8.00 a.m., 10.00 a.m. and noon before and after dexamethasone. Additionally the cortisol response to ACTH was tested under both conditions. The ACTH concentrations in the morning show a periodicity quite similar to the well known circadian rhythm of cortisol secretion. Dexamethasone blocks this periodicity without decreasing ACTH significantly. The cortisol response to ACTH after dexamethasone is normal. In the morning only a basal and presumably biologically inactive ACTH is secreted which can not further be suppressed by dexamethasone. Therefore it is advisable for the diagnosis of hypercortisolism to measure ACTH only in stimulation tests, e.g. insulin-induced hypoglycemia. PMID- 6281631 TI - [Indications for surgery in liver tumors]. PMID- 6281632 TI - [Captopril in refractory advanced cardiac insufficiency]. PMID- 6281633 TI - [Negative effects of dietary bulk]. PMID- 6281634 TI - [Various aspects of the regulation of aldosterone secretion]. AB - Angiotensin was regarded as the major stimulus of aldosterone secretion until the late sixties, when the discussion about the regulation of aldosterone release has been renewed. The present study demonstrates the response of plasma renin activity, aldosterone in serum and electrolytes in serum and urine to ACTH (0.25 mg Synacthen i.v.), pretreatment with dexamethasone (2 mg Millicorten p.o.) and to combined administrations of furosemide (40 mg Lasix i.v.) + potassium (80 mmol Kalinor p.o.) and furosemide + propranolol (5 mg Dociton i.v.) in eight healthy volunteers. Plasma renin activity and aldosterone were measured by radioimmunoassay, electrolytes in serum and urine according to routine methods. ACTH increases aldosterone without influencing plasma renin activity. Pretreatment with dexamethasone decreases aldosterone and stimulates renin response to upright posture. Potassium delays the increase of plasma renin activity after furosemide and propranolol inhibits the furosemide-induced renin release, both without impairing aldosterone secretion. We conclude that the regulatory system of aldosterone production is far more complex than suggested by the concept of the renin-angiotensin-aldosterone-system. PMID- 6281635 TI - Drugs for treatments of systemic fungal infections. PMID- 6281636 TI - [Hepatitis B antigen in the serum in chronic liver diseases and malignant hepatoma]. PMID- 6281637 TI - Higher order chromatin structure determines double-nucleosome periodicity of DNA fragmentation. AB - Double-nucleosome periodicity of DNA fragmentation with DNAse I in the nuclei of cells differing in size of the linker DNA length and lysine-rich histone composition was analyzed by means of nondenaturing agarose gel electrophoresis. DNAse I revealed this type of periodicity in rat thymus and CHO cell nuclei as well as in erythrocyte nuclei. It has been deduced that the so-called nucleodisome structure is also typical of cells possessing a usual DNA repeat length (200 bp or less) and lysine-rich histone H1. Two probably related events are important for establishing a clear double-nucleosome periodicity of DNA fragmentation: the replacement of H1 histone by a specific arginine-rich histone fraction (H5 histone in the case of erythrocyte) and the increase of the linker DNA length. The results are interpreted in terms of supranucleosomal organization of chromatin which may determine the dinucleosome periodicity of DNA fragmentation due to a specific packing of nucleosomes. PMID- 6281638 TI - The role of histone H2B from sea urchin sperm in the association of reconstituted minichromosomes. AB - A comparative study of the condensation of reconstituted complexes of circular SV40 DNA with core histones from calf thymus and sea urchin sperm was performed using sedimentation and electron microscopic techniques. It is shown that in low ionic strength solutions both types of complexes are similar to native 'minichromosomes'. In the region from 0.08 to 0.16 M NaCl the complexes of SV40 DNA with thymus histones form small compact particles. By contrast, the compaction of the SV40 DNA complexes with sperm histones results in the formation of giant intermolecular associates. The results obtained may mean that histone H2B of sea urchin sperm participates in the formation of a higher order structure in sperm chromatin. PMID- 6281639 TI - A longitudinal study of respiratory viruses and bacteria in the etiology of acute otitis media with effusion. AB - We analyzed data from a 14-year longitudinal study of respiratory infections in young children to determine the relative importance of viral respiratory infection and nasopharyngeal colonization with Streptococcus pneumoniae and Haemophilus influenzae as factors influencing the occurrence of acute otitis media with effusion. The incidence of this disorder was increased in children with viral respiratory infections (average relative risk, 3.2; P less than 0.0001). Infection with respiratory syncytial virus, influenza virus (type A or B), and adenovirus conferred a greater risk of otitis media than did infection with parainfluenza virus, enterovirus, or rhinovirus. Colonization of the nasopharynx with Str. pneumoniae or H. influenzae had a lesser effect on the incidence of the disease (average relative risk; 1.5; P less than 0.01). Infections with the viruses more closely associated with acute otitis media (respiratory syncytial virus, adenovirus, and influenza A or B) were correlated with an increased risk of recurrent disease. Prevention of selected otitis associated viral infections should reduce the incidence of this disease. PMID- 6281641 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 23-1982. A 37-year-old woman with headaches, memory loss, and an abnormal CT scan. PMID- 6281640 TI - Evidence that hepatitis B virus has a role in liver-cell carcinoma in alcoholic liver disease. AB - We compared the presence of serologic markers of hepatitis B virus (HBV) infection with the presence of the viral DNA in the livers of patients with alcoholic liver disease with or without hepatocellular carcinoma. Among 51 patients with various kinds of alcoholic liver disease but without hepatocellular cancer, 19 had one or more serologic markers of HBV, but only three had viral surface antigen in their serum. These three patients, as well as three others who had HBV antibodies but no viral antigen in their serum and two others who had no serologic markers of any kind, had HBV DNA in their liver cells. In at least five of the eight patients with viral DNA in the liver, the DNA was integrated into the genome. Among 20 patients with alcoholic cirrhosis and hepatocellular carcinoma, nine of the 16 tested had serologic markers of HBV infection, but all 20 had HBV DNA integrated into the genome of the neoplastic liver cells. These data suggest that HBV plays a part in the pathogenesis of primary liver-cell cancer in alcoholics. PMID- 6281642 TI - Discordant cortisol response to exogenous ACTH and insulin-induced hypoglycemia in patients with pituitary disease. PMID- 6281643 TI - Sounding boards. After laetrile, what? PMID- 6281644 TI - Extrapancreatic, extraintestinal gastrinoma: effective treatment by surgery. PMID- 6281645 TI - Molecular epidemiology of antibiotic resistance in salmonella from animals and human beings in the United States. AB - We collected serotyped isolates of salmonella from reference laboratories in the United States, tested their susceptibility to antibiotics, and extracted plasmids from isolates that were resistant to a different combination of antibiotics from each of three serotypes. Restriction-endonuclease digestion showed that within each of the three groups, plasmid molecules from animal and human isolates were often identical or nearly identical. One serotype-plasmid combination appeared to be endemic in cattle in 20 states and infected 26 persons in two states. The human cases, which were not recognizably related except for their common plasmids, appeared to be clustered in time but geographically dispersed, like cases in previous outbreaks spread by food products. These findings suggest that resistance plasmids may be extensively shared between animal and human bacteria, and that spread of multiresistant strains of salmonella among animals and human beings, as observed in Britain, may have been undetected in the United States for lack of comparable surveillance. PMID- 6281646 TI - Looking at genes. PMID- 6281647 TI - Cytotoxic t cells in cytomegalovirus infection: HLA-restricted T-lymphocyte and non-T-lymphocyte cytotoxic responses correlate with recovery from cytomegalovirus infection in bone-marrow-transplant recipients. AB - We studied 58 recipients of bone-marrow transplants to evaluate immune responses to cytomegalovirus infection. Such infection developed in 43 patients; it was fatal in 12, nonfatal in 23, and present at death from other causes in eight. All patients had low or absent cytomegalovirus-specific cytotoxic lymphocyte activity before the onset of infection. Cytomegalovirus-specific cytotoxic responses developed in all survivors, whereas only two patients with fatal infection had even low-level cytomegalovirus-specific cytotoxic responses. Natural and antibody dependent killer-cell activities were depressed both before and during infection in patients with fatal infections, but not in those who survived. The outcome of the infection did not correlate with the nature of the underlying disease, the type of transplant received, the pretransplantation cytomegalovirus-antibody status, or lymphocyte-proliferation responses to cytomegalovirus antigens or concanavalin A. The correlation between effective virus-specific cytotoxic response and recovery from infection indicates that these effector cells probably mediate recovery from cytomegalovirus infection. PMID- 6281648 TI - Free radicals in arylamine carcinogenesis. AB - Free radical processes and their involvement in carcinogenesis is an unresolved question but one subjected to intense investigation recently. Using in vitro systems, we demonstrated that certain heme compounds combined with hydroperoxides catalyzed the oxidation of N-hydroxy-2-fluorenylacetamide (N-OH-2-FAA) to nitroxyl free radical intermediate which dismutated to form 2-nitrosofluorene (2 NF) and N-acetoxy-2-fluorenylacetamide (N-AcO-2-FAA). Ascorbate and certain antioxidants inhibited this reaction. Lipid hydroperoxides were effective substrates for this reaction, especially in target tissue, rat mammary gland parenchymal cells. Cytochrome P420 in the high-spin state catalyzed the reaction effectively but low-spin cytochrome P420 or P450 were ineffective. Recently, we found that 2-NF added covalently to unsaturated carbon-carbon double bonds of membrane lipids to form an adduct termed 2-nitroso-fluorene lipid adduct (N-O LAF), which, in it oxidized state, exists in the membrane as a nitroxyl free radical. This N-O-LAF undergoes reduction-oxidation changes in the natural membrane. Formation of N-O-LAF occurred in rat liver microsomal membranes by deacylation of N-OH-2-FAA, but the esterase inhibitor paraoxon, prevented its formation from N-OH-2-FAA. The mutagenicity of 2-NF was enhanced in Salmonella typhimurium TA98 if the bacteria were cultured to contain more unsaturated membrane lipids. However, synthesized adducts were only slightly mutagenic. PMID- 6281649 TI - Enzymatic mechanisms of DNA repair. AB - DNA repair proficiency in cells is expressed by various enzymes which can recognize damaged sites arising from exogenous agents or endogenous conditions. Either a damaged base is recognized by DNA glycosylases, partially removed by hemi-DNA glycosylases acting on diadduct damage, or direct incision of the phosphodiester bond near the damaged site. Incision at those apurinic or apyrimidinic sites arising from depurination-depyrimidination or glycosylase reactions is effected by apurinic or apyrimidinic endonucleases. Excision of damaged sites is catalyzed by unique exonucleases followed by DNA polymerase catalyzed reinsertion of nucleotides. The integrity of the strands is restored by polynucleotide ligase when a juxtaposed nucleotide is properly reinserted. PMID- 6281650 TI - Rival transmitter in visual transduction. PMID- 6281651 TI - Expression of two proteins from overlapping and oppositely oriented genes on transposable DNA insertion element IS5. PMID- 6281652 TI - Effect of chloroform on charge movement in the nerve membrane. PMID- 6281653 TI - Adenosine diphosphate induces binding of von Willebrand factor to human platelets. PMID- 6281654 TI - Structural analysis of insertion sequence IS5. PMID- 6281655 TI - The "fold-back"elements of Drosophila. PMID- 6281656 TI - Avian sarcoma virus Y73 genome sequence and structural similarity of its transforming gene product to that of Rous sarcoma virus. AB - From the complete nucleotide sequence of the genome of the avian sarcoma virus Y73, we have predicted amino acid sequence of p90 gag-yes, the product of the transforming gene. Contrary to previous evidence from molecular hybridization studies p90 gag-yes was found to have much homology with the transforming gene product p60 src of Rous sarcoma virus, suggesting that the cellular counterparts of the two (c-yes and c-src) originated from a common prototype sequence. PMID- 6281657 TI - Acumentin, a protein in macrophages which caps the "pointed" end of action filaments. AB - Macrophages have a calcium-insensitive protein which binds to and inhibits monomer addition at the "pointed" end of actin filaments labelled with heavy meromyosin. This protein, in concert with calcium-sensitive proteins already shown to bind the "barbed" end of filaments, can lead to complete control of actin filament length in the cytoplasm. The blocking of actin filament ends in low as well as high calcium concentrations would prevent treadmilling in living cells. PMID- 6281658 TI - Dimeric tetrapeptide enkephalins display extraordinary selectivity for the delta opiate receptor. PMID- 6281659 TI - N-methyl-C-aspartate-type receptors mediate striatal 3H-acetylcholine release evoked by excitatory amino acids. PMID- 6281660 TI - Isolation and structural organization of the human preproenkephalin gene. PMID- 6281661 TI - Facilitation of microdissection by use of a new microscopic and micromanipulatory unit. PMID- 6281662 TI - Assay of avian leukosis viruses by indirect immunoperoxidase method. AB - Chick embryo fibroblast cells (CEF) infected with avian leukosis viruses were stained selectively by the indirect immunoperoxidase method. Good results were obtained by the use of a successive combination of periodate-lysine paraformaldehyde fixation and diaminobenzidine reaction mixture. Viral antigens were detected type-specifically on infected cells. Type-specific antisera determined by the neutralization test were absorbed by the homologous type of virus-infected CEF, but not by the heterologous type of these cells. This test was more effective for detecting virus infectivity than the resistance-inducing factor test. Viral antigen was observed 2 days after inoculation with a large amount of the virus. The minimum infective dose of the virus for the antigen detection was 100 resistance-inducing units (RIU) per plate 4 days after infection, or 1 RIU per plate in CEF after two passages. PMID- 6281663 TI - Pathological changes in laying hens inoculated with the JPA-1 strain of egg drop syndrome-1976 virus. AB - Rhode Island Red laying hens were inoculated with the JPA-1 strain of egg drop syndrome-1976 virus and examined pathologically 1, 3, 5, 7, 10, 14, 21, 28 and 80 days postinoculation (p.i.). Eggs with a discolored or soft shell or no shell were laid over a period from 10 to 24 days p.i. Egg production failed transiently in some affected birds after 13 days p.i. At autopsy, the uterus revealed remarkable edema and swelling of the mucosal folds together with the deposition of whitish exudate in the cavity 14 days p.i. Microscopically, intranuclear inclusion bodies were found in epithelial cells of the uterus, isthmus and vaginal gland region 10 and 14 days p.i. There were also severe degeneration and desquamation of the epithelial cells, atrophy of the uterine glands and remarkable infiltration of heterophils, lymphocytes and plasmacytes accompanied with extensive edema. Twenty-one days after inoculation, lymphoid follicles were formed in the mucosal folds of some part of the oviduct. Electron microscopic examination revealed various amounts of virus particles 68 approximately 80 nm in diameter mainly in the nuclei of the epithelial cells on the mucosa and in the exudate, and also in the cytoplasm of macrophages in the uterus. PMID- 6281664 TI - Pathogenicity of avian nephritis virus for embryonating hen's eggs. AB - The pathogenicity of avian nephritis virus (ANV) for embryonating hen's eggs was studied by various routes of inoculation. When inoculated with ANV by the yolk sac route, 6-day-old embryos showed the highest susceptibility and all of them died 3 to 14 days postinoculation (PI). They manifested hemorrhage and edema of the whole body (3 to 6 days PI) and stunting (7 to 14 days PI). The 50% egg infective dose of the virus by yolk sac inoculation coincided well with the virus titer expressed in plaque-forming units determined on the monolayer of chicken kidney cell cultures. The virus could be passed serially through the chorioallantoic membrane (CAM) of embryonating hen's eggs. In these eggs the CAM presented edematous thickening at the inoculation site, and the embryo stunting. when inoculated by the CAM route, high virus doses killed all embryos, but low virus doses allowed some of the infected embryos to hatch normally. When inoculated by the allantoic cavity route, the virus did not multiply in the allantoic cavity of embryonating eggs, but some of these eggs became infected. Fluorescent antigens were present only in the kidneys and CAM of embryos infected with the virus. The virus was recovered at a low rate from cloacal swabs of chicks from normally hatched eggs inoculated with the virus by the CAM route. These chicks were variable in growth, but had antibodies against the virus and developed nephritis at 36 days of age. PMID- 6281665 TI - GABA induced changes in acetylcholine release from slices of guinea-pig brain. AB - The effect of GABA on acetylcholine (ACh) release was investigated on superfused slices of guinea-pig cerebral cortex (CC), caudate nucleus (CN), tuberculum olfactorium and brain stem. GABA (1--6 x 10(-3) mol/l) increased the spontaneous and KCl-evoked ACh overflow in CC and CN, reduced the electrically-evoked release in all areas tested (most evidently in CC and CN) and lowered the threshold of electric stimulation-induced ACh release in CC. These effects were also caused by 3-amino-1-propane sulphonic acid (1 x 10(-3) mol/l) and ethanolamine-O-sulphate (2 x 10(-3) mol/l), were reduced by bicuculline (1 x 10(-4) mol/l) and fully antagonized by picrotoxin (8 x 10(-5) mol/l), but they were not influenced by phentolamine, methysergide, spiroperidol or strychnine. Tetrodotoxin (TTX) (5 x 10(-7) mol/l) blocked the facilitation of spontaneous ACh release by GABA only when the slices were perfused with normal Krebs solution, but not when perfused with a KCl-enriched medium. These results suggest that GABA affects the cholinergic transmitter release through bicuculline- and picrotoxin-sensitive receptors, showing low affinity toward the agonist. Moreover GABA modulation of resting ACh release requires action potentials only in normal [K+]0, but not in high [K+]0, suggesting that GABA-receptive sites are located at cholinergic terminals. PMID- 6281666 TI - Hippocampal noradrenergic responses in vivo and in vitro. Characterization of alpha and beta components. AB - Pressure ejection of l-norepinephrine (NE) in the in vivo rat hippocampus generally produced depression of pyramidal cell spontaneous activity. In addition, both excitation and biphasic responses were observed. NE-induced inhibition of firing rate was effectively antagonized by concurrent administration of the alpha antagonist phentolamine, but was largely unaltered by the beta antagonist timolol. On the other hand, NE-induced elevation in spontaneous firing rate was effectively blocked by timolol, and largely unaffected by phentolamine. Another beta antagonist, sotalol, did not selectively antagonize either NE-induced inhibition or NE-induced excitation. The beta agonist 2-fluoro-NE produced increases in pyramidal cell firing rates in most cells studied, while the alpha agonist 6-fluoro-NE inhibited the majority of cells examined. The effects of sotalol were also examined on alpha and beta receptor-mediated field responses in the in vitro hippocampal slice. Sotalol was shown to be a selective beta antagonist in this system, blocking excitation evoked by the beta agonist isoproterenol while having no effect on inhibition elicited by the alpha agonist clonidine; however, the potency of sotalol (Ki = 3.5 microM) was considerably less than that of timolol (Ki = 50 nM). Taken together, these results suggest that NE-induced depression and elevation in hippocampal pyramidal cell spontaneous discharge in vivo are mediated via alpha and beta adrenoceptors, respectively. PMID- 6281667 TI - Interaction between mianserin and clonidine at alpha 2-adrenoceptors. AB - The purpose of the present study was to characterize the effects of mianserin at alpha 2-adrenoceptors. Firstly, the action of mianserin on postganglionic sympathetic fibres has been studied using the tachycardia induced by stimulation of the cardiac nerve in dogs. Mianserin increased this tachycardia, but could not prevent the inhibitory effect of clonidine in this model. However, an antagonistic effect of mianserin against clonidine was observed when animals were pretreated with desipramine. Secondly, mianserin antagonized the inhibitory effect of clonidine on the electrically stimulated guinea-pig ileum. In high concentrations, mianserin reduced both electrically and acetylcholine induced contractions. Thirdly, mianserin antagonised the sleep induced by clonidine in chickens. These results are consistent with alpha 2-adrenoceptor blocking properties of mianserin in peripheral noradrenergic fibres in dogs, in cholinergic fibres in guinea-pig ileum and in the central nervous system in chickens. PMID- 6281668 TI - The inhibition by dopamine of cholinergic transmission in the isolated guinea-pig ileum. Mediation through alpha-adrenoceptors. AB - 1. Segments of the guinea-pig ileum were incubated in Tyrode's solution containing 3 microM propranolol. Dopamine, like noradrenaline and clonidine, inhibited the twitch response to field stimulation. The inhibitory action of dopamine remained unchanged in the presence of the dopamine uptake inhibitor nomifensine (1 microM). Tissue from reserpine-pretreated animals was insensitive to tyramine but the response to dopamine was not affected. It is, therefore, assumed that the effect of dopamine is due to a direct receptor stimulation and not to the release of noradrenaline. 2. The inhibitory action of dopamine was not antagonized by the dopamine receptor antagonists cis-flupenthixol, pimozide or domperidone. 3. Metoclopramide, sulpiride and tolazoline were competitive antagonists of the inhibitory effects of dopamine. The pA2-values for metoclopramide against dopamine (5.64), noradrenaline (5.52), and clonidine (5.57) did not differ significantly. Likewise, there was no significant difference between the pA2-values for sulpiride (5.30, 5.29, 5.50) and those for tolazoline (6.52, 6.69, 7.02) determined against dopamine, noradrenaline and clonidine. 4. Apomorphine inhibited the twitch response, and this inhibition was not affected by tolazoline, sulpiride or pimozide. 5. It is concluded that dopamine inhibits the twitch response in the guinea-pig ileum through stimulation of neuronal alpha-adrenoceptors. Metoclopramide and sulpiride are weak antagonists at these receptors. The results provide no evidence for the existence of specific inhibitory dopamine receptors in the guinea-pig ileum. PMID- 6281669 TI - Characterization of autonomic receptors in the rat sublingual gland by biochemical and radioligand assays. PMID- 6281670 TI - Binding characteristics of (+)-, (+/-)- and (-)-[125iodo] cyanopindolol to guinea pig left ventricle membranes. PMID- 6281671 TI - Ouabain sensitivity of Rb+ uptake in canine Purkinje fibre and ventricular muscle. AB - Ouabain inhibitable active Rb+ uptake was compared in canine Purkinje fibre and the left ventricular muscle to define the possible difference in electrophysiological sensitivity to ouabain in the two tissues. Ouabain inhibitable, active uptake of Rb+ was calculated from the difference between total Rb+ uptake assayed in the absence of ouabain and nonspecific Rb+ accumulation assayed in the presence of 10(-4) mol/l ouabain. A similar degree of active Rb+ uptake was inhibited by ouabain in both Purkinje fibre and the left ventricular muscle from the same animal, even though the Purkinje fibre took up more 86Rb+ than the ventricle. The specific tissue binding of ouabain in vitro was also not significantly different in the functionally different two tissues. It is concluded that there is no differential ouabain sensitivity of Rb+ uptake in canine Purkinje fibre and ventricular muscle. PMID- 6281672 TI - Asymmetric release of cyclic AMP from guinea-pig and rabbit gallbladder. AB - The release of cyclic adenosine 3':5'-monophosphate (cAMP) from guinea-pig and rabbit gallbladder was investigated in vitro. Serosal addition of prostaglandin E1 (PGE1) to luminally perfused guinea-pig gallbladders caused a concentration dependent efflux of cAMP to the mucosal side, the threshold concentration of PGE1 being 10(-7) M. The efflux of cAMP to the serosal side was 7-fold lower. A mucosal sidedness of cAMP release was also observed in stripped preparations of rabbit gallbladder mucosa mounted between two half chambers. No cAMP was found in the solutions bathing the serosal layers isolated from rabbit gallbladders. Fluid secretion was observed at 10(-7) M PGE1, an effect mimicked by serosal, but not mucosal application of cAMP (3.3 x 10(-3) M). This is taken to indicate that the basolateral membrane is more easily permeated by cAMP than the apical membrane, since cAMP is believed to exerts its physiological effects from inside the cell. It is concluded that preferential release of cAMP to the mucosal side is not due to a higher permeability of the brush border membrane but rather represents an as yet undefined transport process which may be of importance for the regulation of excessive intracellular cAMP levels. PMID- 6281673 TI - alpha-Adrenoceptors mediating the positive inotropic effect of phenylephrine in the right ventricular muscle of the monkey (Macaca fuscata). AB - The property of adrenoceptors mediating the positive inotropic effect (PIE) in the ventricular muscle of the Japanese monkey (Macaca fuscata) was investigated by the use of phenylephrine (PE) and adrenoceptor antagonists. The intrinsic activity (0.6) and the pD2-value (5.41) for PE were comparable to those in other mammalian species. The beta-adrenoceptor antagonist, pindolol (3 x 10(-8) mol/l) shifted only the upper part of the concentration-response curve (CRC) for PE to the right; the pD2-value for PE was not significantly affected by pindolol. On the other hand, phentolamine (10(-6) mol/l) shifted the lower part of the CRC for PE more than the upper part. In the presence of both pindolol and phentolamine the curve was shifted to the right in a parallel manner. The time required for twitch relaxation was negatively correlated to the degree of PIE of PE in the presence of phentolamine but not pindolol. These results indicate that both beta- and alpha-adrenoceptors mediate the positive inotropic action in the ventricular muscle of the Japanese monkey and that in contrast to the action via beta adrenoceptors the action via alpha-adrenoceptors is not accompanied by the "relaxant effect". PMID- 6281674 TI - [Clostridium perfringens sepsis following criminal abortion]. PMID- 6281676 TI - [The impact of surgical treatment on endocrinological status of pituitary adenomas associated with hyperprolactinemia (author's transl)]. PMID- 6281675 TI - [Antibodies for DNA and RNA viruses in the serum of myasthenia patients. Preliminary report]. AB - The reported material included 14 patients with myasthenia and 14 healthy controls. Serum samples were taken for investigations twice at intervals of several weeks. The titres of antibodies against adenoviruses, cytomegalovirus and herpes virus were determined in the complement fixation reaction using antigens from tissue cultures of viruses. The level of antibodies against influenza viruses A1, A2, and B, rubella nad measles viruses was determined using the test of haemagglutination inhibition. In the myasthenic patients serological investigations demonstrated a higher level of antibodies against the studied viruses as compared with controls. The greatest differences were observed in the levels of antibodies against cytomegaloviruses. In 12 patients cytomegalovirus antibodies were found in serum dilutions from 1:4 to 1:64.. On the other hand in the control group in 5 cases positive results were obtained and the titres of antibodies ranged from 1:4 to 1:16. These data are in agreement with the results of investigations reported by American authors who put forward the hypothesis that persistent cytomegalovirus could be a factor releasing autoimmune reactions of the thymus. PMID- 6281677 TI - Evidence for inhibition by beta-endorphin of vasopressin release during foot shock-induced stress in the rat. AB - This study was to ascertain the effect of naloxone and dexamethasone on vasopressin and beta-endorphin release in the rat during inescapable electric foot shock stress. Plasma vasopressin concentrations were not affected by electric foot shock in vehicle-treated rats, but were raised significantly by the stress in animals pretreated with naloxone. The stress-induced increase in plasma beta-endorphin-like immunoreactivity (beta-EI) was similar whether the rats had received naloxone or not. Plasma beta-EI consisted of equal amounts of beta endorphin-like and beta-lipotropin-like material as revealed by gel filtration. Dexamethasone almost abolished the foot shock-induced increase in plasma beta-EI and, in the presence of dexamethasone, stress was now effective in elevating plasma vasopressin concentrations. These results are consistent with the hypothesis that beta-endorphin, released from the anterior pituitary, inhibits the release of vasopressin from the posterior lobe of the pituitary gland during foot shock-induced stress. PMID- 6281678 TI - Significance of ACTH4-10 in the control of hippocampal corticosterone receptor capacity of hypophysectomized rats. AB - The effect of hypophysectomy on the number of corticosterone receptor sites was investigated in three rat brain regions and was compared with the effect of long term adrenalectomy. Subsequently, the effect on receptor capacity was measured after the hypophysectomized rats had received as substitution therapy ACTH1-24 and smaller peptide fragments lacking corticotropic activity. All rats (sham and hypophysectomy) were adrenalectomized 24 h prior to sacrifice for depletion of endogenous adrenal hormones and replacement therapy was discontinued at that time. Extensive perfusion with saline was carried out at the time of sacrifice. 3H-corticosterone binding was measured in cytosol by means of an in vitro assay. 2 weeks after hypophysectomy, the apparent maximal binding capacity (Bmax) of the corticosterone receptor system was increased by 60, 36 and 72% in hippocampus, hypothalamus and septum, respectively. The increase in Bmax in the hippocampus of hypophysectomized rats was 30% higher than the increase in animals adrenalectomized 2 weeks previously. Replacement with ACTH1-24 markedly decreased the binding capacity in all brain regions investigated. Replacement with the behaviorally active ACTH4-10 sequence reduced the number of corticosterone receptor sites in the hippocampus by 21%, while the behaviorally inactive ACTH11 24 sequence was ineffective. Des-glycinamide-arginine-vasopressin was also ineffective. There were no alterations in binding affinity for corticosterone in hippocampal cytosol after the surgical procedures or after the different replacement therapies. It is concluded that the neurotropic ACTH4-10 sequence reduces the number of corticosterone receptor sites in the hippocampus of the hypophysectomized rat. The action of ACTH4-10 was specific for the hippocampus and was not observed in other brain regions or plasma transcortin. PMID- 6281679 TI - A long-term follow-up study of 214 children with the syndrome of infantile spasms. AB - The long-term prognosis of 192 surviving children with the syndrome of infantile spasms was evaluated. The children had been admitted to three paediatric hospitals in Helsinki at the time of initial diagnosis. The aetiological factors of the syndrome were carefully studied in each case. ACTH therapy was employed in 162, usually for about six weeks. The follow-up study 3-19 (mean 10.4) years later was made at the Children's Hospital, University of Helsinki. The rate of mortality was 19.6 per cent. Normal development was seen in 12 per cent and slightly subnormal in 10.4 per cent of the surviving children. Psychiatric disorders were seen in 27.6 per cent of the survivors. Sensory defects were also common. Severe cerebral palsy was seen in 4 per cent. Other seizures after cessation of the infantile spasms were seen in 60 per cent. Serial EEG studies showed that the temporal lobe was the most common site of abnormality. Abnormalities in the temporal lobes were seen frequently in children with symptomatic neonatal hypoglycaemia as a probable cause of the spasms. Prognostically favourable factors were "idiopathic" aetiology, normal development and not other fits prior to the spasms, short treatment lag, good response to ACTH and short duration of the spasms. In this study early treatment seemed to be of great importance even with regard to mental development. The factors connected with a bad outcome were: symptomatic aetiology (especially brain malformations, early infections and tuberous sclerosis), slow development before spasms, other seizures before infantile spasms, early onset of the spasms, long treatment lag, long duration of the spasms and other later occurrence of myoclonic-astatic seizures (Lennox-Gastaut). Large doses of ACTH (120-160 units) were not associated with a better prognosis than the smaller doses (20-40 units). The benefit of long versus short treatment schedules could not be evaluated in this study. The relapse rate here was 32 per cent. PMID- 6281680 TI - Response of kidney and bone to parathyroid hormone in children receiving anticonvulsant drugs. AB - The response of kidney and bone to parathyroid extract (PTE) was investigated in 8 epileptic children on long-term treatment with primidone in combination with phenytoin or other anticonvulsant drugs. The results indicate a dissociation between normal and cyclic AMP excretion and disturbed renal handling of phosphate which resembles type II pseudohypoparathyroidism suggesting an anticonvulsant drug related inhibition of cyclic AMP-induced phosphaturia. It is speculated that antiepileptic drugs may provoke renal conservation of phosphate which may explain the relative low incidence of manifest rickets or osteomalacia in site of low 25 hydroxy-vitamin D levels in epileptic patients. A normal bone response of PTE indicates that antiepileptic treatment with phenobarbital and phenytoin does not affect PTH-stimulated bone resorption in the investigated patients. PMID- 6281681 TI - Effect of serotonin depletion induced by p-chloroamphetamine on changes in rats' activity levels-produced by lithium. AB - The role of serotonin (5-HT) transmission in the production of changes in rats' activity levels induced by 0.15 and 1.50 mEq/kg LiCl was assessed by selectively depleting 5-HT brain levels with p-chloroamphetamine prior to chronic administration of LiCl for 21 days. While p-chloroamphetamine pretreatment decreased 5-HT levels by 56%, irrespective of the fact that the animals had been administered either saline, 0.15, or 1.50 mEq/kg LiCl for 21 days, it did not alter the effects that each LiCl dose alone that on activity levels. Both LiCl administrations produced an 18% increase in 5-HT levels in rats pretreated with saline, yet diametrically opposed effects on activity levels. A parallel is drawn between these biochemical findings and previously reported data indicating that both LiCl doses produce an improvement in selective attention, yet opposed effects on rats' activity levels. The hypothesis that 5-HT transmission might be crucially involved in mediating the dose-independent effect of lithium on selective attention is proposed for future research. PMID- 6281682 TI - Independent release of supranormal acetylcholine quanta at the rat neuromuscular junction. AB - This electrophysiological study deals with the occurrence and with the mode of release of unusually large miniature end-plate potentials at the rat neuromuscular junction during physiological conditions. A specific limit for the normal miniature end-plate potential amplitude at each cell studied was determined after fitting the observed frequency-amplitude histogram to a Gaussian distribution. The relative abundance of giant miniature end-plate potentials was 4.15% at room temperature. The occurrence of giant miniature end-plate potentials was temperature dependent. The percentage of giant miniature end-plate potentials was 5.8% and 0.61% at 35 degrees C and at 16 degrees C, respectively. The amplitude-independence of the intervals between miniature end-plate potentials was demonstrated at room temperature as well as at 35 degrees C and at 16 degrees C. The results of this study show that giant miniature end-plate potentials are produced by acetylcholine packets which are released independently and that they are not a consequence of the synchronous release of several normal-sized quanta. Moreover, the results indicate that during physiological conditions a minor but regular proportion of the spontaneous release of acetylcholine is made up of larger packets, which produce miniature end-plate potentials of supranormal amplitude. PMID- 6281683 TI - Correlation between acetylcholine release and neuronal activity in the guinea-pig ileum myenteric plexus; effect of morphine. PMID- 6281684 TI - Development of sympathetic ganglionic neurotransmission in the neonatal rat. Pre- and postganglionic nerve response to asphyxia and 2-deoxyglucose. AB - To determine the time course of development of neurotransmission in the sympathetic ganglion of the rat, pre- and postganglionic activity was recorded from the cervical sympathetic trunk in anesthetized neonatal and mature preparations. Tonic activity and responses to two stimuli, cellular hypoglycemia induced by 2-deoxyglucose and asphyxia, which are known to evoke CNS-mediated sympathetic activation in mature rats were measured. In 2-11-day-old neonates, tonic preganglionic activity recorded from the cervical sympathetic nerve and responses to hypoglycemia and asphyxia were comparable to or greater than that in mature rats. In 17-19-day-old neonates these variables were elevated to twice the adult value. In contrast, tonic postganglionic activity recorded from the internal carotid nerve was barely detectable through 5 days of age and there was no response to hypoglycemia. During asphyxia, maximum postganglionic impulse frequency and total number of impulses discharged were 10-20% of the mature value through the 5th postnatal day and the duration of the postganglionic response was only 25% of the preganglionic response. Tonic postganglionic activity and response to stimuli were equivalent to those in mature rats by the 10th postnatal day. The compound action potential evoked in the postganglionic axons by direct electrical stimulation was comparable in 4-5 and 10-13 day-old rats. In the concluded that functional ganglionic neurotransmission is established in the neonatal rat between the 5th and 10th postnatal day. The relation between biochemical changes associated with maturation of the postganglionic neuron, ganglionic synaptogenisis and neurotransmission is discussed. It is concluded that synaptogenisis and onset of neurotransmission are causally associated with development of CNS regulation of postganglionic activity and end organ response rather than with maturation of the postganglionic neuron and that cholinergic excitation of the postganglionic neuron adequate to evoke action potentials is not essential to initiate maturation of the neuron. PMID- 6281685 TI - [Typification of the immunoproliferative syndromes]. PMID- 6281686 TI - [Lymphadenitis caused by Epstein-Barr virus and cytomegalovirus]. PMID- 6281687 TI - [Hormones and breast cancer. Current views]. PMID- 6281688 TI - [Meaning and development of markers of acute hepatitis]. AB - The Authors analyse the results of their tryls about the markers of HBV and HAV acute hepatitis (HBsAg, anti-HAVAb) by R.I.Z. method. HAVAb was in 75% of the cases, its meaning was of post-contact with HAV. The title of HAVAb was effected in 18 patients with viral acute hepatitis; the results were 3 cases of HAV acute hepatitis and in other 7 cases no Ano B viral acute hepatitis. The 58.3% of acute viral hepatitis was HBsAg positive, the study of other markers of HBV and the title of HAVAb showed a viral acute hepatitis caused by HBV. We were not able to demonstrate the viruses which caused 7 HBsAg negative acute virale hepatitis, anti-HBcAg was in 97.8% of HBV acute hepatitis; its the most sensitive of HAV past-infection. The system c-anti-e was in 78.2% of HBsAg viral acute hepatitis. The persistence after 7th week of illness of HBeAg coincided with the hepatitis cronicity. On the contrary anti-HBeAg has not always a protective meaning. PMID- 6281689 TI - [Hepatitis B and hepatitis A markers in 33 cases of acute hepatitis B surface antigen-negative, hepatitis A virus antibody-positive viral hepatitis. Importance of immune complexes]. AB - The authors have looked for the markers of HBV by R.I.A. method (HBsAg, anti HBsAg, HBeAg, anti HBeAg, anti HBcAg), of HAV (by measurement on two samples of HAVab or by measurement of HAVab IgM), the immune-complexes (I.C.C.) by C1q solid phase binding assay method with E.L.I.S.A. with determination after division I.C.C. of HBsAg by R.I.A. method in 33 cases of HBsAg negative acute viral hepatitis. The 9% (3 cases) were HAV acute hepatitis, the 42,4% (14 cases) no A no B acute hepatitis, the 36,3% (12 case) were HBV acute hepatitis, in 9 anti HBcAg positive cases the I.C.C. with HBsAg positive after division resulted positive, the 12,3% (4 cases) had a positivity for HAVab by stereoconversion (2 cases) or HAVab IgM (2 cases) with HBsAg positivity after division I.C.C. This result puts a nosologic problem about the 4 cases of acute viral hepatitis, which, from epidemiological and clinical point of the view are HAV acute hepatitis. PMID- 6281690 TI - [Hormone receptors in colonic cancer]. PMID- 6281691 TI - [Behavior of urinary excretion of cyclic AMP in post-menopausal osteoporosis during therapy with salmon calcitonin]. AB - Salmon calcitonin has been administered at a dose of 100 U.M.R.C. for 6 months in 10 patients suffering from post-menopausal osteoporosis. During treatment, calcium plasma levels fluctuated but tended to fall, while urinary excretion of cyclic AMP rose, this pointing to an enhancement of parathyroid function. After 6 months an increase in intestinal calcium absorption and a decrease in bone turnover were also observed, the bone mineral content evaluated by bone densitometry showed a slight quantitative increase but proved to be substantially unchanged. Salmon calcitonin thus proved capable of interfering with bone turnover by reducing the bone resorption. By fostering an increase in parathyroid hormone production, it also made it possible to exploit that anabolic effect which, in low doses, the parathormone exerts on the bone tissue. PMID- 6281692 TI - [2 cases of trophoblastic degeneration in twin pregnancy]. PMID- 6281693 TI - An A1-adenosine receptor, characterized by [3H] cyclohexyladenosine binding, mediates the depression of evoked potentials in a rat hippocampal slice preparation. AB - In slices of rat hippocampus, adenosine and several adenosine derivatives depressed evoked neuronal responses to afferent stimulation. The nanomolar potency of adenosine derivatives and their relative effectiveness indicate that the depression of evoked potentials is mediated via an A1-adenosine receptor. A remarkable similarity was found between the relative potencies of nucleoside derivatives with respect to their electrophysiological effects and to their inhibition of high affinity [3H] cyclohexyladenosine ([3H]CHA) binding to rat brain membranes. We conclude that the [3H] CHA binding site in rat brain membranes represents a physiological receptor of the A1-type. PMID- 6281695 TI - Evidence for functional interactions of morphine in substantia nigra and striatum in relation to muscular rigidity in rats. PMID- 6281694 TI - Caerulein and cholecystokinin octapeptide (CCK-8): sedative and anticonvulsive effects in mice unaffected by the benzodiazepine antagonist Ro 15-1788. AB - Cholecystokinin octapeptide (CCK-8), caerulein and diazepam inhibited exploratory rearing activity and harman-induced convulsions in mice. Pretreatment with the selective benzodiazepine receptor antagonist Ro 15-1788, reduced or abolished the sedative and anticonvulsive effects of diazepam, but left the same effects of both peptides unaffected. The peptide-induced ptosis was even increased by Ro 15 1788. The results suggest that the CCK-like peptides do not directly interact with the benzodiazepine receptor. PMID- 6281696 TI - Effects of edrophonium bromide on neuromuscular transmission in a healthy human subject. AB - Regional intravenous injections of different amounts (1-3 mg) of edrophonium were given to the hand of a healthy subject. The earliest change was a small negative deflection occurring at the end of the muscle response evoked by nerve stimulation, due to repetitive activity. It did not occur after a second stimulus at 30 ms or immediately after 10 s maximum voluntary contraction. Repetitive stimulation at 0.5 Hz reduced the repetitive activity. With a higher dose of edrophonium the response to a second shock (M2) at 30 ms was reduced in amplitude, but M2 at 80 ms was unaffected. An even larger dose caused depression of M2 at 80 ms also and a decremental response to 50 Hz stimulation. The amplitude of the response to a single shock was unchanged throughout. PMID- 6281697 TI - Effect on neuromuscular transmission of repeated administration of an organophosphorus compound, metrifonate, during treatment of children with urinary schistosomiasis. AB - Muscle action potential amplitude recorded from abductor pollicis brevis in response to nerve stimulation was measured in 55 children during treatment of urinary schistosomiasis with metrifonate (3 doses at 2 weekly intervals). Mean erythrocyte cholinesterase activity was 52-75% of pretreatment value in different groups when examined electrophysiologically. Twenty-six children acted as controls. There was no difference in amplitude between control and exposed subjects 2 weeks after the 2nd dose. Six hours after the 3rd dose, amplitude was larger in some subjects. This effect was not related to dose or degree of cholinesterase inhibition and was thought unlikely to be the result of treatment. Three children who received the highest dose of metrifonate had developed repetitive activity 6 hr later. The criteria for its identification are described. PMID- 6281698 TI - Elevated serum angiotensin-converting enzyme in pulmonary granulomatosis. PMID- 6281699 TI - Rotavirus infection in New Zealand. AB - Rotavirus infection is commonly found in young infants admitted to hospital with gastroenteritis. An enzyme-linked immunosorbent assay (ELISA) for virus diagnosis is described and the results of testing stool specimens from 497 children with gastroenteritis, 192 neonates and 247 asymptomatic six month old infants are presented. Rotavirus infection was found in 45 percent of all children with gastroenteritis but only in 4.7 percent of neonates and 2 percent of asymptomatic infants. These results do not support the proposal that children in our community have a high incidence of subclinical infections. PMID- 6281700 TI - The prevalence of antibodies to hepatitis A and B viruses in Port Chalmers residents. PMID- 6281701 TI - Rotavirus infection in Otago: a serological study. AB - A method for measuring rotavirus antibody in human sera has been established using enzyme-linked immunosorbent assay (ELISA). A Simian strain of rotavirus (SA11) was used as the antigen. Serum eluted from dried blood spots on good quality chromatography paper was found suitable for analysis. Paired serum samples from children with gastroenteritis have shown a brisk antibody response in association with the presence of rotavirus in the faeces. Community studies indicate that although all older children and adults tested have detectable antibodies to rotavirus, there is a significant rise in the number of individuals with high titre antibody in the child bearing age group, after which the levels diminish. This finding suggests that repeated infections occur throughout childhood and early adult life. PMID- 6281702 TI - Hydatidiform mole and gestational trophoblastic disease in Southern Connecticut. AB - Of 127 patients with hydatidiform mole in southern Connecticut, 34 (28%) received chemotherapy for persistently elevated human chorionic gonadotropin (hCG) titers. An hCG regression curve was found to be useful if not mandatory for following patients. Excess uterine size, theca lutein cysts, uterine bleeding, and histologic trophoblastic hyperplasia were relative discriminators of the need for chemotherapy. In the absence of metastases, an hCG titer was the only valid discriminator for initiating chemotherapy, provided the patient could be followed consistently and reliably. The indications for initiating chemotherapy are discussed. Early diagnosis and close follow-up were associated with low morbidity. Five of 6 patients with metastatic disease were referred from outside the center. PMID- 6281703 TI - Suture materials currently used in obstetric-gynecologic surgery in the United States: a questionnaire survey. AB - Questionnaires on the material and the size of sutures used in obstetric gynecologic surgery were sent to the directors of 110 residency programs in the specialty in medical schools in the United States. Replies were received from 80 (73%) programs. The findings indicate that the use of chromic catgut still predominates, but that the newer synthetic absorbable materials such as polyglycolic acid and polyglactin 910 are becoming popular for some procedures, including certain steps in abdominal closure and in reconstructive tubal surgery. The increasing use of these materials for hysterectomy and cesarean section, heretofore exclusively the domain of chromic catgut, suggests that there is possibly under way a reorientation of attitudes about the materials used for sutures in obstetric-gynecologic surgery. The characteristics of catgut and of synthetic absorbable materials are herein described and compared in relation to their current use. PMID- 6281704 TI - Use of intravenous Premarin in the treatment of dysfunctional uterine bleeding--a double-blind randomized control study. AB - Dysfunctional uterine bleeding results from disruption of the normal sequence of ovarian cyclic hormonal stimulation of the uterine endometrium. Treatment of dysfunctional uterine bleeding previously consisted of surgical or hormonal manipulation, or both. The present study was conducted on 34 patients in a randomized double-blind fashion to evaluate the efficacy of treating dysfunctional uterine bleeding with intravenous Premarin. Bleeding stopped in 72% of patients who received intravenous Premarin and in 38% who received placebo (P = .021). Premarin was effective in terminating endometrial bleeding in patients with biopsy-proved pathology consisting of secretory, proliferative, menstrual, polypoid, or cystic hyperplasia, as well as endometritis. PMID- 6281705 TI - Antenatal diagnosis of sickle cell disease: amniotic fluid cell DNA analysis. AB - Using recombinant DNA methods and amniocentesis, the antenatal diagnosis of sickle cell disorders can be accomplished. This article presents the authors' experiences with 21 pregnancies at risk using HpaI and Hind III restriction enzyme digestions of adult leukocytic and fetal amniotic fluid cell DNA. The authors were able to establish direct beta A- and beta S-globin gene linkages to the restricted fragments in 9 families. In 8 families no direct linkages could be determined and estimates of exclusion of fetal beta S homozygosity were used. These estimates were made on the basis of the frequencies of association of the HpaI fragments with the beta S and beta A genes in the Afro-American population. In 4 families, no estimates or linkages could be established, and the patients were counseled in reference to fetoscopy. The data also indicated the frequencies of association of the HpaI fragments with the beta A and beta S gene in the New Jersey-New York population. These studies did not seem to indicate any exclusive preferential segregation of the HpaI and Hind III polymorphisms. PMID- 6281706 TI - Infection as a predominant cause of perinatal mortality. AB - During a 15-month period, all 34 infants delivered at the department of obstetrics and gynecology at University Hospital in Lund, Sweden, who died perinatally or neonatally were included in a prospective study of causes of death. Autopsies--including extensive culturing of specimens for bacteria, chlamydia, fungi, mycoplasmas, and viruses--were performed for all infants. Maternal sera obtained during pregnancy and after delivery were examined regarding titers against a number of microorganisms. During the study period, the perinatal mortality rate was 0.60% and the neonatal mortality rate 0.56%. It was found that 37% of the deaths were caused by lethal malformations, 17% by idiopathic respiratory distress syndrome, and 9% by ablatio placentae. However, no less than 21% occurred as a direct consequence of infections, including 2 deaths caused by group B streptococci, 2 by Coxsackie B virus, and 3 deaths each by Hemophilus influenzae, Pseudomonas pyocyanea, and Candida albicans. A 6-month study of late abortions revealed another case of intrauterine group B streptococcal infection. The study has demonstrated that autopsy, including microbial examination, is recommended in all cases of perinatal and neonatal deaths. PMID- 6281707 TI - Biologic effects of various doses of ethinyl estradiol in postmenopausal women. AB - To determine which dosage of estrogen might provide physiologic replacement while minimizing adverse effects, 20 postmenopausal women were studied before and after oral administration of ethinyl estradiol. Twenty premenopausal women studied in the early and late follicular phases of the menstrual cycle were presumed to reflect normal physiologic function. Variable responses of the different biochemical and biologic markers to the actions of ethinyl estradiol were observed. Liver protein synthesis was the most sensitive measure of the action of ethinyl estradiol. In comparing the relative potencies of ethinyl estradiol with previously reported results observed with the usage of conjugated equine estrogens, the actions of 10 micrograms ethinyl estradiol were approximately equivalent to the biologic effects of 1.25 mg conjugated estrogens. The results suggest that ethinyl estradiol is far more potent than previously believed and that the daily administration of 10 micrograms, a dose lower than currently available commercial preparations, may be adequate for relief of symptoms of vaginal atrophy and may provide protection from the occurrence of osteoporotic fractures. PMID- 6281708 TI - [T-immunity system study in adenovirus infection of the eye]. PMID- 6281709 TI - Children's brain tumour cells produce RNA particles with incomplete retrovirus characteristics. AB - Short-term cultures of cells from human rain tumours have been reported to synthesise RNA particles of density in the range characteristic of C type RNA retroviruses, with associated DNA polymerase activity. Fresh tumour cells obtained from 6 children with astrocytoma and 7 children with medulloblastoma, together with one sample of normal brain tissue and normal leukocytes from brain tumour patients were assayed by several characteristics for the primate retrovirus. 1 or 6 (17%) astrocytomas and 4 of 7 (57%) medulloblastomas released RNA particles which banded in sucrose gradients at a density of 1.16-1.18 g/cm3 together with a short segment of DNA, which was eliminated by prior ribonuclease treatment and two proteins of 28k and 16k daltons. These findings were compatible with the presence of a primate retrovirus. Immune coprecipitation of 125I labelled proteins from the 1.16-1.18 g/cm3 gradient region failed to show any reactivity with antisera to p28 core antigens or the p70 reverse transcriptase antigens of simian sarcoma virus, baboon endogenous virus or Mason Pfizer virus. Assays for DNA polymerase activity in culture supernatant fluid showed only a low amount of activity with template preferences not characteristic of the retroviral reverse transcriptase enzyme. PMID- 6281710 TI - CAMP differences between clones of high and low malignant fibrosarcoma cells. AB - The levels of 3',5'-cyclic adenosine monophosphate (cAMP) were measured in a high malignant and a low malignant clone of murine fibrosarcoma cells isolated from a common parent in a normal mouse fibroblasts at various stages of growth. There was an inverse correlation between the degree of malignant potential and the cAMP levels. The normal fibroblasts had the highest cAMP at all time periods measured. The high malignant cells had the lowest levels and the low malignant cells were intermediary. The degree of susceptibility of each population to the growth inhibiting effects of dibutyryl cAMP, a cAMP analogue, was also measured. There was a direct correlation between degree of malignant potential and susceptibility to growth inhibition. These findings strongly support our belief that the low malignant cells represent a state of malignancy that is intermediary between the high malignant cells and the normal fibroblasts. cAMP differences may contribute to this. PMID- 6281711 TI - Monomorphic adenomas of the major and minor salivary glands. Report of twenty-one cases and review of the literature. AB - Monomorphic adenomas are benign salivary gland tumors that have a predilection for development in the upper lip and parotid gland. Typically, patients are older persons (mean age, 61 years), but a broad age range (32 to 87 years) has been reported in the literature. Adequate treatment consists of superficial or total parotidectomy (depending on extent and location of the tumor) for parotid lesions and excision with a limited border of normal tissue for minor gland tumors. Uniform cellularity, lack of myxoid or chondroid features, and a tendency for multicentric origin are features which separate these tumors from pleomorphic adenomas. Monomorphic adenomas have been mistakenly diagnosed and treated as adenoid cystic carcinomas. Close attention to cytologic detail, histomorphology, and growth pattern at the periphery are important in separating these tumors. PMID- 6281712 TI - [Does receptor memory exist and is it inherited?]. PMID- 6281713 TI - [A case of benign hepatocellular adenoma]. PMID- 6281714 TI - [Gas phlegmon of the neck and chest (author's transl)]. PMID- 6281715 TI - The relevance of ultrastructural examination in the classification of primary lung tumours. AB - Biopsies from 50 primary lung tumours were classified according to the World Health Organisation's Histological Typing of Lung Tumours. They were also subjected to electron microscopic examination. Comparison of the diagnoses made by these separate methods showed that many poorly differentiated squamous cell carcinomas had been incorrectly classified. Agreement was good in the diagnosis of adenocarcinoma whilst ultrastructural examination of small anaplastic carcinomas disclosed a neuroendocrine tumour with a combination of squamous and glandular elements. Large cell anaplastic carcinoma proved to be a 'waste-basket' containing tumours which displayed ultrastructural characteristics of poorly differentiated squamous cell carcinoma, poorly differentiated adenocarcinoma or neuroendocrine carcinoma. Electron microscopy was also valuable in characterization of other pulmonary tumours whose identity could not be resolved at the light microscopic level. Ultrastructural examination may provide a better understanding of the histogenesis and derivation of lung tumours, as well as their behaviour and therapeutic response. PMID- 6281716 TI - Haemorrhagic manifestations with Sindbis infection. Case report. AB - Sindbis infection in man occurs rarely in Australia. Most recorded cases are either asymptomatic or result in a fever sometimes accompanied by a macular or vesicular rash. This case is of particular interest because of the severe haemorrhagic vesicular rash and the repeated recurrence of symptoms over a 5 month period together with the persistence of IgM antibodies to Sindbis virus. PMID- 6281717 TI - Rotavirus viral RNA electrophoresis in hospitalized infants with diarrhea in Santiago, Chile. AB - Viral RNA electrophoresis technique was used to detect rotavirus in 226 children under 2 years of age with acute diarrhea, admitted to the Roberto del Rio Hospital in Santiago, Chile, during the period of June 1979 through May 1980. A group of 50 children included in the aforementioned sample, admitted in winter, was compared with a control group of 25 infants without digestive pathology. In these groups, rotavirus was detected in 20 out of 50 children with diarrhea (40%) but not in the controls (0%). A positive diagnosis of rotavirus was found in 66 out of the total of 226 patients (29.2%); its monthly distribution ranged between a maximum of 83.3% (June) and a minimum of 11.1% (October). PMID- 6281718 TI - Congenital cytomegalovirus infection: diagnostic and prognostic significance of the detection of specific immunoglobulin M antibodies in cord serum. AB - Specific immunoglobulin M antibodies were detected by radioimmunoassay (RIA-IgM) in cord sera from 83/93 (89%) babies congenitally infected with cytomegalovirus (CMV) but in 0/104 cord sera from uninfected control subjects. The type of maternal infection did not affect the ability of the assay to identify congenital infections, but increased RIA-IgM titers were found more frequently in cord sera from babies infected following primary CMV infections (9/18; 50%) than following recurrent CMV infections (1/12; 8%) (P less than .05). The magnitude of the fetal immune response was related to disease inasmuch as 14/40 (35%) babies with increased RIA-IgM titers were symptomatic at birth compared with 1/43 (2%) with lower titers (P less than .001). When combined with the results of testing for rheumatoid factor and total IgM, the RIA-IgM assay defined subgroups of babies with generally poor (7/15; 47% symptomatic at any stage) or generally good (0/21 symptomatic) prognoses. Prospective studies currently identifying cases of congenital CMV infection may wish to use these three serologic techniques as the results obtained appear to have prognostic significance for those babies who are initially asymptomatic. PMID- 6281719 TI - Child health and social status. AB - Available evidence regarding the relationship between socioeconomic status and health in childhood has been summarized. Only studies that used income, education, or occupation as measures of socioeconomic status and provided data obtained subsequent to legislation facilitating access to care have been cited. Data are presented on the prevalence and severity of illness (mortality, acute conditions as a group, chronic conditions as a group, and hospitalization), sequelae of prematurity, and specific common health problems and their sequelae. These specific health problems are lead poisoning, vision problems, otitis media and hearing loss, cytomegalic inclusion disease, asthma, psychosocial and psychosomatic problems, and iron deficiency anemia. All of the above (with the possible exception of asthma) are more prevalent among poor children than among nonpoor children. Even more striking is the evidence for consistently greater severity of problems or likelihood of sequelae among poor children. Although causality cannot be inferred from these data, the findings suggest a need for more basic research on the social correlates of disease, on the effect of social progress on disease prevalence and severity, and on the effect of medical care in overcoming the disadvantage associated with low socioeconomic status. PMID- 6281721 TI - Posttransfusion cytomegalovirus infections. PMID- 6281720 TI - Defects of tooth structure in congenital cytomegalovirus infection. PMID- 6281722 TI - [Cis-platinum-induced peripheral neuropathy. One case (author's transl)]. AB - A case of peripheral neuropathy secondary to cis-platinum therapy is reported. On the basis of electrophysiological studies and of biopsies, the predominantly sensory disorders observed (paraesthesias, pain, sensory ataxia) could be ascribed to axonal degeneration. This exceptional complication of cis-platinum therapy deserves to be borne in mind, since the drug is increasingly used, notably in genital cancers. PMID- 6281723 TI - [Evaluation of methods of diagnosing breast cancer]. PMID- 6281724 TI - [Pseudohypertriglyceridemia due to hyperglycerolemia caused by glycerol kinase deficiency]. PMID- 6281725 TI - The DNA sequence of the gene rpsA of Escherichia coli coding for ribosomal protein S1. AB - The DNA sequence of the gene rpsA as well as of its neighboring regions has been determined using the dideoxyribonucleotide method. It was found that there is an "open-reading-frame" of 350 bp which precedes the gene rpsA. Furthermore, an extensive internal repeats of nucleotide sequence have been found in this gene. PMID- 6281726 TI - The nucleotide sequence of the bacteriocin promoters of plasmids Clo DF13 and Co1 E1: role of lexA repressor and cAMP in the regulation of promoter activity. AB - Treatment of cells, harbouring the bacteriocinogenic plasmic Clo DF13 with mitomycin-C, which induces the cellular SOS response, results in a significantly increased transcription of the operon encoding the bacteriocin cloacin DF13, the immunity protein and the lysis protein H. The nucleotide sequences of the promoter regions and N-terminal parts of the bacteriocin genes of Clo DF13, Col E1 and the pMB1 derivative pBR324 have been determined. A comparison of these sequences with those of corresponding regions of the lexA, recA and uvrB genes revealed that the promoter regions of the bacteriocin genes studied contain binding sites for the lexA protein, which is the repressor of the E. coli DNA repair system. Using both, a thermosensitive lexA host strain and a host with pACYC184 into which the lexA gene had been cloned, we were able to demonstrate, that in vivo the lexA protein is involved in the regulation of bacteriocin synthesis. From the data presented, we conclude that bacteriocin synthesis is controlled at least by the lexA repressor. It has been reported that also catabolite repression might play an essential role in the control of bacteriocin synthesis. Computer analysis of the DNA sequence data indicated that the promoter regions of both, the cloacin DF13 and colicin E1 genes contain potential binding sites for the cyclic AMP-cyclic AMP Receptor Protein complex. PMID- 6281727 TI - Cloning and determination of the transcription termination site of ribosomal RNA gene of the mouse. AB - A Eco RI 6.6 kb DNA fragment containing the 3'-end of 28S ribosomal RNA gene of the mouse was detected by Southern blot hybridization, and cloned in a lambda phage vector. The site of transcription termination and the processed 3'-end of 28S RNA were determined on the cloned fragment and the surrounding nucleotide sequence determined. The 3'-terminal nucleotides of mouse 28S RNA are similar to those of yeast, Drosophila and Xenopus although the homology was lost drastically beyond the 3'-end of 28S RNA. 45S precursor RNA terminated at 30 nucleotides downstream from the 3'-end of 28S RNA gene. A structure of a dyad symmetry with a loop was found immediately prior to the termination site of 45S RNA. The rDNA termination site thus shares some common features with termination sites recognized by other RNA polymerases. PMID- 6281728 TI - Isolation and characterization of a human collagen alpha 1(I)-like gene from a cosmid library. AB - We have isolated a human collagen alpha 1(I)-like gene from a cosmid library. The clone which contains 37kb of human DNA has been shown to contain this gene by DNA sequencing, hybrid arrest and hybrid selection assays and Northern blot hybridizations. The collagen gene sequence extends through most of the cloned DNA and must, therefore, be at least 35kb in length. PMID- 6281729 TI - New procedure using a psoralen derivative for analysis of nucleosome associated DNA sequences in chromatin of living cells. AB - Sites for restriction endonuclease cleavage in double helical DNA are blocked from cleavage when the photoaffinity drug trimethylpsoralen is photobound at or near the site. In general, Hind III sites are about 15 fold more sensitive to inactivation than the other restriction sites which were tested, although sensitivity of different Hind III sites seems to vary somewhat depending on base sequences adjacent to the site. Hind III sites can be inactivated in two ways; one which completely blocks action of the specific restriction endonuclease and one permitting the introduction of a swivel which relaxes DNA supercoiling without producing a double strand break. Nucleosomes and perhaps other protein DNA complexes can protect the underlying DNA sequence from trimethylpsoralen photobinding and thus protect restriction sites from inactivation. This property can be exploited to determine if specific sites are accessible to the psoralon probe in vivo and thus to establish if specific nucleotide sequences are nucleosome associated. Using this procedure evidence is obtained that nucleosomes on SV40 DNA in living infected cells are either distributed randomly or at many discrete alternate sites that approach a random distribution. PMID- 6281730 TI - New rapid methods for DNA sequencing based in exonuclease III digestion followed by repair synthesis. AB - We describe improve enzymatic methods for sequencing method for sequencing DNA. They are based on partial digestion of duplex DNA with exonuclease III to produce DNA molecules with 3' ends shortened to varying lengths, followed by repair synthesis to extend and label the 3' ends. After asymmetrical cleavage of the DNA with a restriction enzyme, the labeled products are separated by gel electrophoresis and the sequence read from the autoradiogram. The entire procedures, beginning with unrestricted DNA and followed through gel electrophoresis, takes only one day for sequencing both strands of the DNA molecule. These methods are especially suitable for sequencing DNA cloned in plasmid vectors, and they greatly extend the usefulness of the dideoxynucleotide chain termination method of Sanger et al. (Proc. Natl. Acad. Sci. USA 74, 5463, 1977). Using these methods we have determined the sequence of a 410 base pair fragment which includes the yeast SUP3 tyrosine tRNA gene. PMID- 6281731 TI - Nucleotide sequence of human influenza A/PR/8/34 segment 2. AB - The nucleotide sequence of RNA segment 2 of human influenza strain A/PR/8/34 has been determined. Segment 2 in 2341 nucleotides long and encodes a protein of 757 amino acids (86,500 daltons molecular weight) which is involved in RNA synthesis. Although segment 2 is identical in size to segment 1, which encodes a protein of related function, neither the nucleotide sequences of these two RNA segments nor the amino acid sequences of the encoded proteins appear to be homologous. The sequence of segment 2 completes the sequence of the virus (total 13,588 nucleotides). PMID- 6281732 TI - The construction of cosmid libraries of eukaryotic DNA using the Homer series of vectors. AB - We have constructed and characterised a series of approved, disabled cosmid vectors which we call Homer cosmids and have examined the optimal conditions for the construction of libraries of eukaryotic DNA segments using these vectors. Analysis of these libraries shows that most of the sequences we have tested for are present at the expected frequency and that the libraries can be stably propagated. We have also directly tested the stability of cosmid clones carrying tandemly repeated inserts. This work shows that it should be possible to clone most eukaryotic genes using cosmid vectors and that such cloning systems have considerable advantages over those more commonly used. PMID- 6281733 TI - Infectivity and structure of molecular clones obtained from two genetically transmitted Moloney leukemia proviral genomes. AB - The Mov-2 and Mov-10 substrains of mice, each carrying Moloney leukemia virus (= M-MuLV) in their germ line at the Mov-2 and Mov-10 locus, respectively, do occasionally at a later age (Mov-2) or not at all (Mov-10) activate infectious virus. The M-MuLV proviruses with flanking mouse sequences corresponding to the Mov-2 and Mov-10 locus, respectively, were molecularly cloned. Restriction enzyme analysis revealed no major deletions or insertions in the proviral genomes of the Mov-2 and Mov-10 locus. Both cloned DNAs induced XC plaques in a transfection assay. The specific infectivity, however, was very low and 3T3 cells transfected with the Mov-2 or Mov-10 clone did not produce infectious virus. Removing part of the 5' cellular sequences from the Mov-10 clone did not increase the infectivity. The results suggest that the M-MuLV integrated at the Mov-2 and Mov-10 locus carry a mutation which prevents synthesis of infectious virus but permits XC plaque induction by partial genome expression or synthesis of non-infectious particles. PMID- 6281734 TI - An adenovirus agnogene. AB - The nucleotide sequence of a 550 base pairs long segment, located between map positions 21 and 22.5 in the adenovirus type 2 genome has been determined. S1 nuclease mapping and sequence analysis of cDNA copies of adenovirus mRNA demonstrated that the established sequence includes the i-leader which is spliced to the 5'-end of certain adenovirus mRNAs (1). The i-leader which is 440 nucleotides long contains an open translational reading frame which is preceded by an AUG triplet and which terminates in the third segment of the tripartite leader. A polypeptide with the same molecular weight as predicted from the DNA sequence was identified by in vitro translation of mRNA which had been selected by hybridization to DNA fragments, containing sequences from the i-leader. The results thus suggest that the i-leader, unlike other adenovirus leader segments, is used for translation. PMID- 6281735 TI - A generalized method of subcloning DNA fragments by restriction site reconstruction: application to sequencing the amino-terminal coding region of the transforming gene of Gazdar murine sarcoma virus. AB - The technique of restriction site reconstruction was generalized so as to allow the subcloning of any DNA fragment and its subsequent reexcision with EcoRI, XbaI, XhoI or HindIII. After excision, the 3' terminus of each strand will be derived from the starting nucleic acid, permitting the use of such fragments as primers for nucleotide sequencing by primer extension methods. The technique was used to subclone a 56 base pair BstNI-DdeI fragment of Moloney murine sarcoma virus (Mo-MSV) as a unique HindIII-HindIII fragment. This fragment then served as a primer to sequence a portion of the RNA genome of Gazdar murine sarcoma virus (Gz-MSV). The nucleotide sequence which was obtained indicated that the transforming gene of Gz-MSV arose by at least two recombination events involving murine leukemia virus (MLV) and the cellular homologue c-mos. This analysis suggests that a virus indistinguishable from Mo-MSV was an intermediate in the formation of Gz-MSV. PMID- 6281736 TI - Recognition sites of eukaryotic DNA topoisomerase I: DNA nucleotide sequencing analysis of topo I cleavage sites on SV40 DNA. AB - Eukaryotic DNA topoisomerase I introduces transient single-stranded breaks on double-stranded DNA and spontaneously breaks down single-stranded DNA. The cleavage sites on both single and double-stranded SV40 DNA have been determined by DNA sequencing. Consistent with other reports, the eukaryotic enzymes, in contrast to prokaryotic type I topoisomerases, links to the 3'-end of the cleaved DNA and generates a free 5'-hydroxyl end on the other half of the broken DNA strand. Both human and calf enzymes cleave SV40 DNA at the identical and specific sites. From 827 nucleotides sequenced, 68 cleavage sites were mapped. The majority of the cleavage sites were present on both double and single-stranded DNA at exactly the same nucleotide positions, suggesting that the DNA sequence is essential for enzyme recognition. By analyzing all the cleavage sequences, certain nucleotides are found to be less favored at the cleavage sites. There is a high probability to exclude G from positions -4, -2, -1 and +1, T from position -3, and A from position -1. These five positions (-4 to +1 oriented in the 5' to 3' direction) around the cleavage sites must interact intimately with topo I and thus are essential for enzyme recognition. One topo I cleavage site which shows atypical cleavage sequence maps in the middle of a palindromic sequence near the origin of SV40 DNA replication. It occurs only on single-stranded SV40 DNA, suggesting that the DNA hairpin can alter the cleavage specificity. The strongest cleavage site maps near the origin of SV40 DNA replication at nucleotide 31-32 and has a pentanucleotide sequence of 5'-TGACT-3'. PMID- 6281737 TI - Conservation of high efficiency promoter sequences in Saccharomyces cerevisiae. AB - The position of the yeast phosphoglycerate kinase (PGK) gene has been mapped on a 2.95kb Hind III fragment. We have determined the nucleotide sequence of the 5' flanking region and compared this sequence with those from 16 other yeast genes. PGK, like all other yeast genes has an adenine residue at position -3. It has two possible TATA boxes at positions -114 and -152 and a CAAT box at -129. In addition we have defined a structure at position -63 to -39 that is common to all yeast genes that encode an abundant RNA. This structure is a CT-rich block followed, about 10 nucleotides later, by the sequence CAAG. PMID- 6281739 TI - Nucleotide sequence of the 16S - 23S spacer region in an rRNA gene cluster from tobacco chloroplast DNA. AB - The nucleotide sequence of a spacer region between 16S and 23S rRNA genes from tobacco chloroplasts has been determined. The spacer region is 2080 bp long and encodes tRNAIle and tRNAAla genes which contain intervening sequences of 707 bp and 710 bp, respectively. Strong homology between the two intervening sequences is observed. These spacer tRNAs are synthesized as part of an 8.2 kb precursor molecule containing 16S and 23S rRNA sequences. PMID- 6281738 TI - The chromosomal origin of replication (oriC) of Erwinia carotovora. AB - The chromosomal DNA replication origin (oriC) of the plant pathogen Erwinia carotovora has been isolated and sequenced. The minimal E. carotovora oriC regional functional in Escherichia coli is a 374 base pair region located on a 7.9 kilobase pair SalI fragment which also contains a functional asnA gene. Differences between the nucleotide sequence of the minimal origin regions of E. carotovora and those of E. coli and Salmonella typhimurium are clustered nucleotide substitutions, with regions of complete homology, up to 19 base pairs long, between the three origins. Nine GATC sites are found in the minimal origin, and all are conserved. In contrast, the region toward asnA from the minimal origin shows little clustering and the differences occur mainly every third nucleotide, suggesting that this region is a protein coding region. PMID- 6281740 TI - Correlation of thermodynamic and genetic properties in the Tn10 encoded TET gene control region. AB - The thermal stability of the Tn10 encoded tetracycline resistance (TET) gene control region is investigated by melting studies using purified DNA restriction fragments containing various amounts of flanking sequences. In order to study the thermodynamic properties of this control region under conditions, where enough flanking DNA is present to mimic the situation in the chromosome, the five step melting process of a 1450-bp DNA fragment is analyzed. Because most of the sequence of this DNA is not known, the assignment of the melting transitions to segments of the DNA is done by an experimental method. This employs the preparation of subfragments from the 1450-bp DNA and comparison of their denaturation profiles with the one of the intact sequence. This approach results in the complete assignment of the five denaturation steps. Rather than from the ends, the unwinding starts from the TET gene control region in the middle of the 1450-bp sequence. A clear correlation between the thermodynamic and genetic properties of this DNA is observed. The regulatory sequence forms a small cooperative unit with the lowest stability in the entire fragment. The thermal denaturation of the TET repressor. TET operator complex reveals, that the TET repressor specifically recognizes the double stranded TET operator DNA and stabilizes this structure by 2.4 degrees C. This results is also discussed as an example of the possible action of denaturing or stabilizing proteins on this genetic control region. PMID- 6281741 TI - Metabolic and distribution studies with radiolabeled 5-fluorouracil. AB - 5-Fluorouracil (5-FU) is an effective anti-tumor drug, which has been used both as a single agent and in combination with other chemotherapeutic agents for the treatment of tumors such as breast and colorectal carcinoma. We synthesized 5-FU with trace amounts of 18F-5-FU and administered the compounds intravenously to 6 cancer patients. The patients were scanned at 2 hr intervals for 12 hrs and their urine collected whenever possible. We also injected 5-FU with the tracer 18F-5 FU, at pharmacological doses, into non-tumored rats, and sampled their bile and blood for 95 mins post-injection. For comparison, 2-14C-5-FU was injected into non-tumored rats and their bile and blood sampled at the same intervals. Minute quantities of rat bile and serum were analyzed chromatographically by high performance TLC. 5-FU and two of its metabolites (FBAL and FUPA) were identified and quantified by this technique. Both percentage and absolute amounts of 5-FU in the bile follow comparative kinetic patterns. While the liver and the urinary bladder were clearly observable in all 16 patients, the detectability of the gall bladder was correlated to the inverse of the alkaline phosphatase level in the blood. This work suggests that the diversity of the 5-FU metabolism in cancer patients may allow the use of 18F-5-FU as a probe for understanding those individual variabilities in clinical situations. PMID- 6281742 TI - Computerized study with 201T1 of the cold thyroid node. AB - Because of its physical and potassium-metabolic characteristics 201T1 is more suitable than 131Cs for radioisotopic studies of the cold thyroid nodule, with the further diagnostic possibility of quantitatively assessing intranodular behavior for a specific differentiation among different kinds of neoformations. Using a gamma-camera on line with a computer data processing device, sequential scintiscans were recorded for the first 20-30 min after i.v. administration of 15 20 microCi/kg of radiothallium; delayed sequences were taken at 40-60 min if intranodular uptake appeared. A quantitative appraisal was made of the differential 201T1 uptake-ratio between nodule and healthy thyroid tissue (density-index) and the multiparameter analysis of thyroid time/activity curves generated on the relative regions of interest (ROIs). This computerized study, in 120 out of 293 patients submitted to this radiothallium test, has shown a) diagnostic agreement between clinical-histological and radioisotopic findings in 76 out of 79 colloid-cystic or degenerative neoformations, in all 16 malignant and in 23 out of 25 hyperplastic benign nodules; b) significant statistical difference of the density-index in solid versus cystic but not between benign and malignant nodules; c) different 201T1 kinetics behaviour in different kinds of solid thyroid lesions with a satisfactory statistical difference of the radiothallium nodular disappearance-index. PMID- 6281743 TI - [Causative investigations of heparin-induced rise on thyroid hormone serum levels (author's transl)]. AB - Intravascular application of heparin causes a significant rise of free and total thyroid hormone concentrations in serum together with a concomitant drop of the TSH-levels. A causal relationship between these two findings has been firmly established by a number of investigators. The decline of TSH concentrations in serum after heparin injection might just be a physiological reaction of the pituitary gland to the heparin-triggered rise of serum thyroid hormone levels. The present investigations demonstrate a diminution of affinity of thyroid hormone receptors of liver cells. This result confirms the hypothesis that the rise of total thyroid hormone in blood is mainly due to a depletion of liver stores of thyroid hormones. In concordance with clinical findings the binding affinity of cellular pituitary thyroid hormone receptors was found to increase. PMID- 6281744 TI - [Comparison of the clinical effects to the kinetics of prostaglandin (PGF2 alpha), cAMP, cortisol and ACTH behavior in the plasma of bronchial asthma patients treated with triamcinolone acetonide]. PMID- 6281745 TI - Dynorphin (1-13): analgesia, hypothermia, cross-tolerance with morphine and beta endorphin. AB - Intracerebroventricular administration of 20, 40 and 60 nmol of dynorphin (1-13) produced analgesia, as assessed by flinch/jump response to footshock, and hypothermia in the rat. Rats developed tolerance to both the analgesic and thermic effects of the 20 nmol dose of dynorphin. Dynorphin and beta-endorphin showed cross-tolerance with respect to their analgesic but not their thermic effects. Dynorphin and morphine also produced cross-tolerant analgesic effects. Naloxone (10 mg/kg, IP) completely blocked the barrel rolling produced by 20 nmol dynorphin but did not alter its analgesic or thermic effects. PMID- 6281746 TI - Characterization and subcellular localization of GnRH analog binding in rat brain. AB - The binding of a degradation-resistant analog of gonadotropin-releasing hormone, [D-Phe6]GnRH, to rat brain crude particulate preparation was studied. The binding of this analog at 0 degrees C was saturable and Scatchard analysis revealed the presence of 2 binding sites: one with KD = 1.39 x 10(-7) M and Bmax = 265 pmole/mg protein, and another of lower affinity but higher capacity with KD = 5.58 X 10(-6) M and Bmax = 1734 pmoles/mg protein. The binding at 0 degrees C was substantially higher than that obtained at 37 degrees C, due to binding site inactivation processes occurring at 37 degrees C. The binding sites exhibited a considerable degree of specificity for GnRH as unrelated peptides (with the exception of ACTH) display a much weaker affinity than GnRH and GnRH analogs. Subcellular fractionation demonstrated that most of the binding was associated with the mitochondrial fraction. PMID- 6281747 TI - Bombesin reduces food intake of normal and hypothalamically obese rats and lowers body weight when given chronically. AB - Bombesin is a peptide hormone reported to reduce meal size when administered in rats. In the first experiment, synthetic bombesin was injected subcutaneously into normal rats and obese rats with lesions of the ventromedial hypothalamus just prior to the presentation of food. A dose-dependent suppression of meal size occurred for both groups, showing that the peptide has this action in obese as well as normal animals. In a second experiment, a conditioned taste aversion was not formed with a dose of bombesin which suppressed meal size by approximately 50% while the animals did develop an aversion with a dose of LiCl reported to reduce meal size equivalently. In a third experiment, rats were placed on a feeding schedule where they received three 30-min meals each day. After weights had stabilized under this paradigm, bombesin was administered just prior to each meal for six days. The bombesin caused a consistent suppression of meal size when the animals were allowed 30-min meals such that the rats lost weight over the six day period. When this experiment was repeated with 60-min meals apparent tolerance developed to these actions of bombesin. PMID- 6281748 TI - MSH/ACTH4-10: a tool to differentiate between the role of vasopressin in memory consolidation or retrieval processes. AB - MSH/ACTH4-10 induces a dose dependent increase of latency scores during retention of a passive avoidance response, when injected SC prior to retention but not when administered immediately after the learning trial. Intracerebroventricular administration of anti-vasopressin serum immediately after the learning trial or 1 hr prior to retention induces marked deficits in passive avoidance behavior as indicated by low latencies during retention. SC injection of MSH/ACTH4-10 increased latency scores in animals which received anti-vasopressin serum prior to retention, but did not alter latencies in animals, which received anti vasopressin serum after the learning trial. These results suggest that MSH/ACTH4 10 is involved in retrieval processes and is able to differentiate between the effects of vasopressin on memory consolidation and on retrieval. PMID- 6281749 TI - Modulation of spontaneous seizures in the mongolian gerbil: effects of beta endorphin. AB - Intraventricular injection of beta-endorphin (0.1-3 micrograms) into gerbils from the UCLA seizure sensitive strain reduced the incidence and severity of spontaneous epileptiform seizures, both the motor manifestations and the preceding high voltage focal spiking and accompanying seizure activity in the cortical EEG. This "'anticonvulsant" effect of beta-endorphin was prevented by prior administration of naloxone (1 mg . kg-1 IP). These findings suggest that the endogenous opioid peptide may be involved in the normal suppression of the epileptic diathesis in these animals during the interictal periods. PMID- 6281750 TI - The therapeutic effect of concurrent administration of 5-fluorouracil and allylestrenol or hexestrol in small doses on prostatic carcinoma. AB - A trial to reduce the dosage of estrogen or to substitute it by other agents was done as a treatment of prostatic carcinoma. Either the combination of 5-FU (200 mg/day) and allylestrenol (50 mg/day), or 5-FU and hexestrol (20 mg/day) was given to 24 cases of prostatic carcinoma, 4 of whom were relapsed hormonal resistant cases, 14 cases were controlled by large doses of estrogen and 6 were newly treated cases. No progressions were observed following 3 months of therapy. In 5 cases, progression was observed after 6 months. Deteriorated cases increased to 6 cases after 12 months. The effective rate, according to the "vital score," which is proposed as a parameter of the physical condition of patients with prostatic carcinoma, was calculated to be 100% at 3 months, 77% at 6 months, and 71% at 12 months. PMID- 6281751 TI - Human amniochorion: tissue-specific markers, transferrin receptors and histocompatibility antigens. AB - Human amniochorion was studied by immunofluorescence with the use of antisera to human trophoblast, amniotic epithelium and transferrin as well as with monoclonal antibodies to beta 2M and HLA. The results showed that cytotrophoblast of amniochorion, like its counterpart in placenta, manifests a trophoblast-specific membrane marker and lacks beta 2M and HLA. This tissue also did not react with antisera to amniotic epithelium, and unlike its placental counterpart no evidence for transferrin receptors could be obtained. Amniotic epithelium was found to lack trophoblast antigens, beta 2M, HLA and transferrin receptors, but contained a plasma membrane marker not found on any other extra-embryonic tissues. These results show that extra-embryonic cells with characteristic membrane markers fail to manifest histocompatibility antigens, almost as though they were mutually exclusive. This is supported by the observation that transformed amnion epithelial cells lose their unique surface antigens, react with monoclonal anti beta 2M and acquire transferrin receptors, raising the possibility that the histocompatibility neutrality of extra-embryonic cells is maintained by the normal insertion of specific fetal markers into their plasma membranes. PMID- 6281752 TI - Transferrin receptor affinity and iron transport in the human placenta. AB - Transferrin receptor activity has been investigated on isolated human placental syncytiotrophoblast microvillous plasma membrane preparations following 3 M KCl washing of membranes to remove endogenous receptor-bound maternal transferrin. The calculated binding parameters for essentially apo- and diferric human transferrin, and also for rabbit transferrin, were closely comparable (ranges: Ka, 3.5 to 5.0 X 10(7) M-1; n, 2.6 to 4.0 X 10(14)/mg membrane protein). Iron transport in pregnancy is thus unlikely to involve a simple process of transferrin displacement following iron release at the placental trophoblast plasma membrane receptor site. PMID- 6281753 TI - Immunoelectrophoretic evidence that the human syncytiotrophoblast transferrin receptor is identical to a major plasma membrane antigen present throughout pregnancy. AB - Crossed immunoelectrophoresis of solubilized syncytiotrophoblast microvillar proteins has been performed through intermediate gels, containing a monospecific antiserum to transferrin, into gels containing an antiserum raised to purified microvilli from term placentae. This technique resolved the microvillar transferrin-receptor complex into its component proteins. Crossed hydrophobic interaction immunoelectrophoresis confirmed that the transferrin receptor is an amphiphilic integral membrane protein. It is also a major antigen of the microvillar membrane and is present throughout pregnancy. PMID- 6281754 TI - Immunohistochemical demonstration of placental protein 5 in trophoblastic tumours. AB - An immunoperoxidase technique was used for the detection of placental protein 5 (PP5) in tissue specimens from trophoblastic tumours. PP5 was detected in all of 15 cases of hydatidiform mole, in two out of six destructive moles and in one of seven choriocarcinomas. Though the biological importance of PP5 is not clear, its association with inactivation of plasmin suggests a role in the control of placental invasiveness. PMID- 6281755 TI - [Functional and ultrastructural changes in the adenohypophysis in obesity]. AB - The indices of some tropic adenohypophyseal functions of obese patients were compared with ultrastructural reaction of the adequate adenocytes of rats with experimental hypothalamic obesity. It was shown that in obesity significant functional and structural changes occur in adenohypophyseal cells, their extent being considerably associated with the excessive body mass. In pronounced obesity, a decrease in somato- and thyrotropic- and an increase in gonado- and corticotropic adenohypophyseal functions are seen, manifesting in ultrastructural changes of appropriate adenocytes. The results obtained allow a conclusion, that one of the mechanisms of obesity development is the changed secretory potency of adenohypophyseal cells, producing hormones, which exert a substantial influence upon lipogenesis and lipolysis processes. PMID- 6281756 TI - Kinetics of interaction between beta-receptors, GTP protein, and the catalytic unit of turkey erythrocyte adenylate cyclase. AB - The kinetics of turkey erythrocyte membrane adenylate cyclase activation by beta agonists and guanyl-5'-yl imidodiphosphate is explored as a function of the concentration of the GTP regulatory protein and of the catalytic unit. It was found that the overall kinetics of activation is first order and is independent of the concentration of the GTP regulatory unit N, the catalytic unit C, and of hormone over a very wide concentration range. It was established that the rate limiting step does not involve GDP dissociation from the inactive N unit or the association between activated N' and C. Also, it was found that guanyl-5'-yl imidodiphosphate binding occurs in a random fashion and is not hormone dependent. These results enable us to exclude models of the sequential type in which N in its inactive form is bound to receptor R, is released in an active form N' upon hormone activation, and then binds to C, activating the latter. An acceptable model that accounts for all of the data conforms to the original formulation of "collision coupling" in which N is tightly associated to C at all times. PMID- 6281757 TI - Cytosolic malic dehydrogenase activity is associated with a putative substrate for the transforming gene product of Rous sarcoma virus. AB - A cellular protein of apparent Mr 34,000--36,000 was suggested as a possible physiological substrate for the protein kinase (EC 2.7.1.37) activity associated with the transforming gene product of Rous sarcoma virus. We find this protein to migrate with an apparent Mr of 38,000 in NaDodSO4/polyacrylamide gels. It was not separable from cytosolic malic dehydrogenase activity when purified by chromatography on DEAE-Sephacel, hydroxylapatite, poly(A)-Sepharose, and blue Sepharose, by gel filtration, and by isoelectric focusing. The Mr 38,000 protein as well as cytosolic malic dehydrogenase activity focused with a pI of 7.5. In gel filtration experiments, both displayed an apparent native Mr of 68,000. The male dehydrogenase activity contained in homogeneous preparations of the Mr 38,000 protein had a specific activity of up to 130 units/mg of protein. The recovery of the enzyme was 5--10% of the activity in the extract. Antiserum against the Mr 38,000 protein inactivated the malic dehydrogenase activity associated with the Mr 38,000 protein. PMID- 6281758 TI - Human influenza virus hemagglutinin is expressed in monkey cells using simian virus 40 vectors. AB - We have cloned and expressed the hemagglutinin (HA) gene of a human influenza virus (A/WSN/33) in monkey kidney cells by linking it to deleted simian virus 40 (SV40) genomes that contain the entire early gene region, the origin of replication, and late leader sequences. The HA gene (1775 base pairs long) was originally inserted by the dG . dC tailing technique into the multicopy plasmid of Escherichia coli, pBR322, using cDNA made from viral RNA. The cloned gene was further modified by treatment with nuclease Bal 31 to remove the dG . dC tails and some of the untranslated sequences and recloned in E. coli after addition of BamHI restriction endonuclease linkers. A number of SV40 and HA recombinants (SV- HA) were constructed by inserting recloned HA DNA into the late gene region of SV40. The SV--HA recombinants, when complemented in a lytic infection of monkey cells by the helper function of SV40 early deletion mutants expressed influenza HA as detected by immunofluorescence and immunoprecipitation of in vivo-labeled proteins using either heterogeneous anti-influenza rabbit antibodies or monoclonal antibodies against HA. Furthermore, the WSN HA expressed by the SV--HA recombinants was also glycosylated and possessed the same molecular weight (approximately 70,000) as the uncleaved HA of WSN virus in monkey cells. PMID- 6281759 TI - Posttranscriptional control of bacteriophage lambda gene expression from a site distal to the gene. AB - The bacteriophage lambda int gene product, integrase, recombines the phage DNA with the host DNA at specific sites on each to accomplish lysogeny. The int gene is transcribed from two promoters, PL and PI, each regulated positively by lambda proteins. The expression of integrase is also controlled from a site, sib, in the b region of the phage genome. This is a unique regulatory site because it is located distal to the structural gene in relation to the promoters. The expression of int from the PL promoter is inhibited when sib is present. This effect appears to be specific for PL because sib does not cause inhibition of PI dependent int synthesis. lambda mutants that contain alterations in the site have been isolated. Sequence analyses of the mutations reveal single base changes, spanning 37 base pairs (bp) in the b region, some 240 bp beyond the int gene. Another mutant, hef13, which has a phenotype similar to that of sib, introduces a nucleotide change within the same 37-bp region. The sib and hef mutations cluster within a region of dyad symmetry. Regulation of int synthesis by sib occurs after transcription of the int gene. There is no difference in the rate of PL-promoted int mRNA synthesis in either sib+ or sib- phage infections, yet int mRNA is less stable in the sib+ infection. Because RNase III host mutants are defective in sib regulation, processing of the PL mRNA at sib by this endoribonuclease may cause int mRNA decay and decrease int synthesis. PMID- 6281760 TI - Efficient expression of Escherichia coli galactokinase gene in mammalian cells. AB - The Escherichia coli galactokinase gene (galK) was inserted into a modified early region transcription unit of simian virus 40 (SV40) contained on a bacterial plasmid. Introduction of this pSVK vector into monkey, mouse, and hamster cell lines by transfection resulted in efficient expression of the bacterial galK gene. This expression was shown to be dependent upon fusion of the galK gene to the early promoter of SV40 and did not appear to require SV40 splice signals. Moreover, expression in these cells could be obtained either transiently, 24--72 hr after transfection, or continuously, after stable transformation. In particular, pSVK-dependent galK expression was obtained in a hamster cell line genetically deficient in galactokinase activity. Expression of the bacterial enzyme was shown to complement the galactosemic defect of these cells, thereby allowing their selective survival and growth on galactose as the only carbon source. The ability to readily assay, select for, and potentially select against galK expression from pSVK and its derivatives should prove extremely useful in studying eukaryotic gene regulatory signals. PMID- 6281761 TI - Factors determining frequency of plasmid cointegration mediated by insertion sequence IS1. AB - We demonstrate that mutants with deletions at either end of the insertion sequence IS1 lose the ability to mediate cointegration of two plasmids, whereas mutants with deletions or an insertion within IS1 can mediate cointegration at a reduced frequency. These results, together with the nucleotide sequence analysis of the IS1 mutants, indicate that the two ends of IS1 (insL and insR) and two genes (insA and insB) that are encoded by IS1 are required for cointegration. Using a plasmid carrying two copies of IS1, we found that the individual IS1s mediate cointegration at different characteristic frequencies, and that each of two parts of plasmid DNA segments flanked by the two IS1s is a transposon, mediating plasmid cointegration at a unique frequency. When one IS1 was replaced with a mutant IS1, the remaining wild-type IS1 complemented the cointegration ability of the mutant IS1 as well as a resulting mutant transposon that was then flanked by a wild-type IS1 and a mutant IS1. The efficiency of this complementation reflected the characteristic ability of an individual IS1 present on the plasmid to promote cointegration. The results suggest that the IS1-encoded proteins are produced in different amounts, depending on the location of IS1 in the plasmid, and that these amounts determine the efficiency of complementation of the cointegration ability of a mutant IS1 as well as a mutant transposon. However, the location of an individual IS1 itself can also determine the frequency of cointegration in the presence of a given amount of the IS1 proteins. On the basis of the observation that the cointegration ability of a mutant IS1 is less efficiently complemented than is the ability of a mutant transposon, we also suggest that the IS1-encoded proteins can function in trans, but act preferentially on the IS1 or transposon sequence from which they are produced in promoting cointegration. PMID- 6281762 TI - Regulation of 6-phosphofructo-2-kinase activity by cyclic AMP-dependent phosphorylation. AB - Addition of glucagon to isolated rat hepatocytes resulted in inhibition of 6 phosphofructo-2-kinase (ATP:D-fructose-6-phosphate-2-phosphotransferase) activity in extracts of the cells and in a decrease in the intracellular level of fructose 2,6-bisphosphate. The effect on 6-phosphofructo-2-kinase was characterized by a decrease in the affinity of the enzyme for fructose 6-phosphate. To investigate the mechanism of action of glucagon, 6-phosphofructo-2-kinase from rat liver was partially purified by polyethylene glycol precipitation, DEAE-cellulose chromatography, (NH4)2SO4 fractionation, Sephacryl S-200 gel filtration, DEAE Sephadex chromatography, and Sephadex G-100 gel filtration. Incubation of the purified enzyme with the catalytic subunit of the cyclic AMP-dependent protein kinase from rat liver and [gamma-32P]ATP resulted in 32P incorporation into a protein with a subunit Mr of 49,000 as determined by NaDodSO4 disc gel electrophoresis. Associated with this phosphorylation was an inhibition of 6 phosphofructo-2-kinase activity that was also characterized by a decrease in the affinity of the enzyme for fructose-6-phosphate. Both the phosphorylation and the inhibition of the purified 6-phosphofructo-2-kinase were blocked by addition of the heat-stable protein kinase inhibitor. It is concluded that the glucagon induced decrease in fructose 2,6-bisphosphate levels observed in isolated hepatocytes is due, at least in part, to cyclic AMP-dependent phosphorylation and inhibition of 6-phosphofructo-2-kinase. PMID- 6281763 TI - Gene order and gene-polypeptide relationships of the proton-translocating ATPase operon (unc) of Escherichia coli. AB - We have constructed an extensive set of plasmids that carry the genes specifying the eight polypeptides of the proton-translocating ATPase of Escherichia coli. Using detailed restriction analysis and in vitro protein synthesis directed by these plasmids, we have established the order of the eight unc genes to be BEFHAGDC and the corresponding polypeptides to be a, c, b, delta, alpha, gamma, beta, and epsilon. These analyses include determining the location of the gene coding for the delta subunit of the F1 portion of the complex. We call this gene uncH. We have now established the gene order and gene-polypeptide relationships of the unc operon. This approach should be of use for study of other multigene bacterial operons, especially those with genes coding for polypeptides with unknown or unmeasurable catalytic activity. PMID- 6281764 TI - Regulation of fructose-6-phosphate 2-kinase by phosphorylation and dephosphorylation: possible mechanism for coordinated control of glycolysis and glycogenolysis. AB - The kinetic properties and the control mechanism of fructose-6-phosphate 2-kinase (ATP: D-fructose-6-phosphate 2-phosphotransferase) were investigated. The molecular weight of the enzyme is approximately 100,000 as determined by gel filtration. The plot of initial velocity versus ATP concentration is hyperbolic with a Km of 1.2 mM. However, the plot of enzyme activity as a function of fructose-6-phosphate is sigmoidal. The apparent K0.5 for fructose-6-phosphate is 20 microM. Fructose-6-phosphate 2-kinase is inactivated by the catalytic subunit of cyclic AMP-dependent protein kinase, and the inactivation is closely correlated with phosphorylation. The enzyme is also inactivated by phosphorylase kinase in the presence of Ca2+ and calmodulin. The phosphorylated fructose-6 phosphate 2-kinase, which is inactive, is activated by phosphorylase phosphatase and alkaline phosphatase. The possible physiological significance of these observations in the coordinated control of glycogen metabolism and glycolysis is discussed. PMID- 6281765 TI - Isolation of an altered form of DNA polymerase I from Escherichia coli cells induced for recA/lexA functions. AB - A novel form of DNA polymerase I (deoxynucleosidetriphosphate:DNA deoxynucleotidyltransferase, DNA nucleotidyltransferase, EC 2.7.7.7) activity has been isolated from Escherichia coli cells that had been activated for expression of the DNA damage-inducible genes. Induction was by treatment of normal cells or cells carrying the spr-51 and tif-1 mutations with nalidixic acid. This activity, DNA polymerase I, seems to be a form of DNA polymerase I because it is insensitive to N-ethylmaleimide, is inhibited by antibody to DNA polymerase I, and does not appear in a polA1 strain. DNA polymerase I activity sediments through sucrose gradients as a broad peak with s20.w = 6.6--10.5, compared with an s20,w = 4.8--5.5 for DNA polymerase I. The fidelity during polymerization reactions of DNA polymerase I is relatively low with a variety of synthetic templates and deoxynucleoside triphosphates, although the enzyme appears to have a normal level of 3' greater than 5' exonuclease. Polymerase I has properties that might implicate it in some form of mutagenic DNA repair. PMID- 6281766 TI - Eukaryotic mRNA capping enzyme-guanylate covalent intermediate. AB - Incubation of HeLa cell mRNA guanylyltransferase (GTP:mRNA guanylyltransferase, EC 2.7.7.50) with [alpha-32P]GTP and a divalent cation in the absence of an RNA acceptor results in the formation of a covalent enzyme-guanylate complex. The complex, after purification by phosphocellulose chromatography, can transfer its bound GMP moiety to pyrophosphate, regenerating GTP, or to the 5'-diphosphate end of poly(A), forming a cap structure G(5')pppA(pA)n. The GMP-polypeptide has a molecular weight of 65,000 and is stable to heating in the presence of sodium dodecyl sulfate. On the basis of the alkali-stable and acid-labile nature of the bond and its susceptibility to nucleophilic attack by hydroxylamine at low pH, the GMP-polypeptide linkage appears to be a phosphoamine bond. After digestion with trypsin, a single GMP-peptide was resolved by two dimensional electrophoresis and chromatography. PMID- 6281767 TI - Fate of receptor and ligand during endocytosis of asialoglycoproteins by isolated hepatocytes. AB - The endocytosis leading to degradation of 125I-labeled asialo-orosomucoid specifically bound to the surface of freshly isolated hepatocytes was monitored as a function of time at 37 degrees C. Experimental values were determined for the rates of internalization, dissociation of the receptor-ligand complex, and degradation of the labeled ligand. Compartmental analysis and computer modeling revealed that the data were compatible with dissociation of ligand from receptor preceding ligand degradation. The rate coefficient for internalization was calculated to be an order of magnitude greater than that for receptor--ligand dissociation. Ligand internalization did not result in concomitant depletion in the total number of cell surface receptors. Our data are taken to indicate that ligand remains associated with the receptor after internalization, that the complex is dissociated prior to degradation, and that new, unoccupied receptors are promptly returned to the cell surface from an internal pool. PMID- 6281768 TI - Sequence organization and genomic distribution of the major family of interspersed repeats of mouse DNA. AB - We have investigated that organization and the distribution of a family of interspersed DNA repeats in the mouse genome. The repeats are at least 5600 base pairs (bp) in size and contain two contiguous BamHI endonuclease fragments, 4000 and 540 bp in size, the larger of which includes a 1350-bp EcoRI fragment studied by previous authors. The repeats are polymorphic in their restriction maps, and represent the major family of interspersed repeats in the mouse genome. The repeats are present almost exclusively in the two light major components of mouse DNA, and the base composition of their large BamHI fragments matches that of those components. The genomic distribution of the repeats is different from that of structural genes, which are present not only in the two light components but also in the two heavy components of mouse DNA. This distribution indicates that the repeats are not involved, at least in any simple way, in the regulation of gene expression. PMID- 6281769 TI - Territorial limits and functional anatomy of the simian virus 40 replication origin. AB - The region at and near the simian virus 40 (SV40) DNA replication origin contains a series of palindromes, a 17-base pair (bp) A + T-rich sequence, three copies of a 21-bp repeat, and two copies of a 72-bp repeat. We have constructed a series of recombinant plasmids containing sequential deletions at the region of SV40 DNA replication origin starting from the end near the repeats. These deletions were introduced by using in vitro and in vivo techniques. The relative replication efficiency of these recombinant plasmids were directly assayed in COS-1 monkey kidney cells capable of providing the tumor antigen necessary for the replication of these molecules. Recombinants lacking both copies of the 72-bp repeat did not exhibit any reduction in replication efficiency. Recombinants lacking the 21-bp repeats showed decreased replication efficiency; the reduction in replication efficiency was proportional to the number of copies of the 21-bp repeat deleted in these recombinants. A recombinant retaining the palindromes at the region of SV40 DNA replication but lacking the A + T-rich sequence and the repeats failed to replicate. Based on these results, the SV40 DNA replication origin is subdivided into two regions, and their boundaries are defined. One of these two regions is a core region containing the 17-bp, 15-bp, and 27-bp palindromes and, quite likely, the 17-bp A + T-rich sequence which are necessary for replication. The other is an auxiliary region that consists of the 21-bp repeats and has a dose-dependent enhancement effect on replication efficiency. PMID- 6281770 TI - Resonance Raman spectra of bacteriorhodopsin's primary photoproduct: evidence for a distorted 13-cis retinal chromophore. AB - We have obtained the resonance Raman spectrum of bacteriorhodopsin's primary photoproduct K with a novel low-temperature spinning sample technique. Purple membrane at 77 K is illuminated with spatially separated actinic (pump) and probe laser beams. The 514-nm pump beam produces a photostationary steady-state mixture of bacteriorhodopsin and K. This mixture is then rotated through the red (676 nm) probe beam, which selectively enhances the Raman scattering from K. The essential advantage of our successive pump-and-probe technique is that it prevents the fluorescence excited by the pump beam from masking the red probe Raman scattering. K exhibits strong Raman lines at 1516, 1294, 1194, 1012, 957, and 811 cm-1. The effects of C15 deuteration on K's fingerprint lines correlate well with those seen in 13-cis model compounds, indicating that K has a 13-cis chromophore. However, the presence of unusually strong "low-wavenumber" lines at 811 and 957 cm-1, attributable to hydrogen out-of-plane wags, indicates that the protein holds the chromophore in a distorted conformation after trans leads to cis isomerization. PMID- 6281771 TI - Gap junctional conductance: comparison of sensitivities to H and Ca ions. AB - One cytoplasmic aspect of the junctional membrane between coupled pairs of Fundulus blastomeres was perfused with solutions of known H and Ca ion concentrations. Conductance of junctional membrane was decreased by either ion. The sensitivity to H ions was about 10,000 times greater than that to Ca ions. The results suggest that junctional conductance can be modulated by changes in H ion concentration near physiological pH, but that unphysiologically high concentrations of Ca ion, such as would be reached only on cell death, are required for comparable changes in junctional conductance. PMID- 6281772 TI - Role of Ca2+ and cyclic AMP in the regulation of the production of prostacyclin by the vascular endothelium. AB - Incubation of primary monolayer cultures of human umbilical vein endothelial cells with buffer, thrombin (0.5 unit/ml), ionophore A23187 (10 microM), arachidonic acid (20 microM), prostaglandin H2 (PGH2) (4 microM) resulted in prostacyclin (PGI2) production in nanomolar quantities to the extent of 36 +/- 2, 276 +/- 13, 485 +/- 32, 533 +/- 22, and 532 +/- 22, respectively, as measured by radioimmunoassay of 6-keto-PGF alpha. Preincubation of the endothelium with 1 mM 8-(N,N-diethylamino)-octyl-3,4,5-trimethoxybenzoate, an antagonist of cytoplasmic Ca2+, or with 4 mM 1-methyl-3-isobutylxanthine (MIX), an inhibitor of cyclic nucleotide phosphodiesterase activity, blocked PGI2 release induced by thrombin or A23187, decreased arachidonic acid-induced release by approximately 50%, but had no effect on PGH2-induced release. Radioimmunoassay of cAMP in the endothelium showed that the basal level (1.85 +/- 0.14 pmol of cAMP per 4.5 x 10(5) cells) was increased by an average of 3.9-fold with 4 mM MIX. PGI2 (0.4 microM) had no significant effect on cAMP levels in the absence of MIX, but caused a 2-fold increase with 4 mM MIX. The findings suggest that: (i) the stimulation of PGI2 biosynthesis is mediated by Ca2+, (ii) increased cAMP inhibits PGI2 production, and (iii) cAMP phosphodiesterase activity modulates PGI2-induced increases in the intracellular concentration of cAMP. PMID- 6281773 TI - Thermosensitive development and tip regulation in a mutant of Dictyostelium discoideum. AB - A thermosensitive developmental mutant of Dictyostelium discoideum identifies a gene product that is nonessential for cell multiplication but is continuously required during aggregation and the period when multicellular mounds are formed. After mounds form a tip, which has the properties of an embryonic organizer, this gene product may be nonessential. Surgical removal of the tip from a polarized developing multicellular structure (the slug) leads to emergence of a new tip at the permissive temperature but not at the restrictive temperature. The mutant continues to develop abnormally when mixed with wild-type cells; therefore, a cell-limited rather than an exchangeable factor is altered. Assays show that the mutant has a thermosensitive defect in chemotaxis toward cAMP. The mutation reduces the number of cell surface cAMP receptors expressed at the restrictive temperature without affecting their dissociation constants or their apparent thermostability. The expression of two developmentally regulated enzymes, N acetylglucosaminidase and cAMP phosphodiesterase, is unaffected by the mutation. PMID- 6281775 TI - Physical map of the white locus of Drosophila melanogaster. AB - The white locus of Drosophila melanogaster is a genetically well-characterized locus, mutations in which alter the degree of pattern of pigmentation of the eyes. Using a previously cloned DNA segment containing a portion of the white locus of a mutant allele, we have cloned and characterized the DNA of a 48 kilobase chromosomal region of the Canton S wild-type strain. We have mapped the positions, relative to restriction endonuclease cleavage sites, of several previously characterized chromosomal rearrangement breakpoints that bracket the while locus. These results define a segment of 14 kilobase that contains all of the white locus sequences necessary for the production of a wild-type eye color phenotype. By conventional criteria, no repetitive sequences are present within this 14-kilobase segment; however, we have identified an extremely weak DNA sequence homology between a portion of this segment and a chromosomal region in the vicinity of the zeste locus. PMID- 6281774 TI - Inverted repeat nucleotide sequences in the genomes of Marek disease virus and the herpesvirus of the turkey. AB - The DNAs of two herpesvirus, the oncogenic Marek disease virus and the serologically related herpesvirus of the turkey, were studied by electron microscopy. On the basis of fold-back molecules observed in single-stranded DNA from both viruses, structures have been derived from the overall nucleotide sequence arrangement in their genomes. Although differing in molecular weight, the genomes of Marek disease virus and turkey herpesvirus are both constructed according to the same plan--two regions of unique nucleotide sequence, each enclosed by inverted repeat sequence. The genome structure of these viruses therefore closely resembles that of herpes simplex virus rather than the biologically more similar herpesvirus Epstein--Barr virus, H. saimiri, and H. ateles. PMID- 6281776 TI - Cell surface properties of HLA antigens on Epstein-Barr virus-transformed cell lines. AB - A number of monoclonal antibodies have been used to investigate the distributions and rates of lateral motion of the HLA-A,B, and-DR antigens on several Epstein- Barr virus-transformed B-cell lines. The lateral diffusion coefficients (D) of fluorescein conjugates of the monoclonal antibodies bound to the cell surface were determined by fluorescence recovery after pattern photobleaching. Ds of HLA A and-B were found to be comparable and of the order of 10(-9) to 10(-10) cm2/sec for each of the seven monoclonal antibodies and four cell lines examined. The HLA antigens appear to be monomeric on the cell surface based on experiments using mixtures of arsanilic acid-conjugated and fluorescein-conjugated antibodies. Four monoclonal antibodies against DR antigens were examined. Two of these, Genox 3.53 and L243, labeled the cell surface uniformly and gave Ds comparable to those obtained for the HLA-A and -B antigens. The other two, DA2 and 2.06, rapidly patched on the cell surface and were immobile. The DA2, L243, and Genox 3.53 antibodies bound outside of the caps formed with the arsanilic acid-conjugated 2.06 antibody and a second-step rhodamine-conjugated rabbit anti-arsanilate antibody. This is consistent with recent biochemical evidence that there are multiple distinct antigens coded for by the HLA-DR region. PMID- 6281778 TI - Physical separation and characterization of two types of benzodiazepine receptors. AB - Two distinct benzodiazepine receptors are solubilized differentially by various detergents. The receptor sites that resist solubilization, designated type I, are most highly concentrated in the cerebellum and corpus striatum whereas the more readily solubilized receptors, type II, are most enriched in the hippocampus. The type I receptors display higher affinity for beta-carboline esters and a triazolopyridazine whereas several benzodiazepines do not differentiate the two receptors. The type I receptors can be solubilized with 2% Triton X-100/1 M NaCl; they retain the same drug specificity as in the particulate state. PMID- 6281777 TI - Mechanism for shifting the phase of a circadian rhythm by serotonin: involvement of cAMP. AB - Serotonin (5-hydroxytryptamine, 5-HT) shifts the phase of the circadian rhythm in the eye of Aplysia. We have examined the role of cAMP in mediating the effects of 5-HT on the rhythm. The phase shifts produced by 5-HT are mimicked by treatments that should increase intracellular levels of cAMP. An analogue of cAMP-8 benzylthio-cAMP, advanced and delayed the rhythm at phases in which 5-HT had similar effects on the rhythm. In addition, two phosphodiesterase inhibitors, Ro 20-1724 and papaverine, caused advance phase shifts where 5-HT advances the rhythm. The phosphodiesterase inhibitors Ro-20-1724 and 3-isobutyl-1 methylxanthine each potentiated the effect of subthreshold doses of 5-HT on the rhythm. The effects of 5-HT and 8-benzylthio-cAMP on the rhythm were nonadditive, indicating that 5-HT and 8-benzylthio-cAMP affect the rhythm through a common pathway. Finally, 5-HT produced large changes (13-fold) in the levels of cAMP in the eye. These results indicate that cAMP mediates the effect of 5-HT on the rhythm. There are two possible roles for cAMP in the circadian system. Either the cAMP system is an intracellular step in an entrainment pathway or it is part of the biological clock. Because 5-HT, 8-benzylthio-cAMP, and three phosphodiesterase inhibitors inhibit impulses from the eye, cAMP may also mediate the inhibition produced by 5-HT, or it might be involved in regulating the frequency of spontaneous impulses throughout the day. PMID- 6281779 TI - Reaction mechanism of mRNA guanylyltransferase from rat liver: isolation and characterization of a guanylyl-enzyme intermediate. AB - Rat liver RNA guanylyltransferase catalyzes a GTP-PPi exchange reaction in the absence of acceptor RNA [Mizumoto, K. & Lipmann, F. (1979) Proc. Natl. Acad. Sci. USA 76, 4961-4965] suggesting that the reaction proceeds through the formation of a covalent guanylylated intermediate. We now present more direct evidence for the existence of the enzyme-GMP intermediate: (i) the enzyme-[32P]GMP intermediate was formed on incubation of rat liver guanylyltransferase with [alpha-32P]GTP and migrated as a single radioactive band with Mr 69,000 on NaDodSO4/polyacrylamide gel electrophoresis, and (ii) the intermediate isolated on gel filtration can transfer its GMP moiety to ppGpCpC-poly(A2,U2,G) to form the capped RNA molecule or it can react with PPi to regenerate GTP. The formation of the intermediate was dependent on Mg2+ and was strongly inhibited by PPi. The addition of pyrophosphatase markedly increased the amount of the intermediate complex. On blue dextran-Sepharose affinity column chromatography, the activity of guanylyltransferase to form an enzyme-[32P]GMP intermediate comigrated with activities of cap formation and GTP-PPi exchange. A phosphoamide type linkage between GMP and enzyme is suggested by its acidlabile and alkali-stable nature and also by the susceptibility to acidic hydroxylamine. These results indicate that the reaction catalyzed by rat liver guanylyltransferase occurs through the following two partial steps: (i) E + GTP in equilibrium E-pG + PPi; and (ii) E-pG + ppN .....leads to GpppN .....+ E. PMID- 6281780 TI - Animal cells dependent on exogenous phosphatidylcholine for membrane biogenesis. AB - A Chinese hamster ovary cell (CHO) mutant (strain 58), defective in CDP-choline synthetase (cholinephosphate cytidylyltransferase; CTP:cholinephosphate cytidylyltransferase, EC 2.7.7.15), is temperature sensitive for growth and contains less than half of the normal amount of phosphatidylcholine under nonpermissive conditions [Esko, J. D. & Raetz, C. R. H. (1980) Proc. Natl. Acad. Sci. USA 77, 5192-5196]. We now report that the addition of 40 microM egg phosphatidylcholine or lysophosphatidylcholine to the medium suppresses the temperature sensitivity of mutant 58 and permits the growth of colonies at the restrictive temperature. Phospholipids with different polar headgroups, lipoprotein-bound phospholipids, sphingomyelin, and glycerophosphocholine do not support prolonged growth at 40 degrees C, whereas phosphatidylcholine analogs such as phosphatidyldimethylethanolamine, D-phosphatidylcholine, and beta phosphatidylcholine are quite effective. A broad range of saturated phosphatidylcholines, especially those with fatty acids 12-18 carbons in length, suppresses the phenotype. Phospholipids containing ether-linked hydrocarbons are ineffective, whereas polyunsaturated phosphatidylcholines are toxic. Residual endogenous synthesis of phosphatidylcholine by the mutant is not stimulated under conditions of phenotypic bypass, but the uptake of exogenous lipid is enhanced considerably compared to the wild type. Our findings demonstrate that exogenous phospholipid can provide at least 50% of the phosphatidylcholine required for membrane biogenesis in animal cells and that uptake of exogenous phospholipids may be regulated. PMID- 6281781 TI - Specificity of receptor-mediated recognition of malondialdehyde-modified low density lipoproteins. AB - Blood-borne human monocytes and macrophages derived from human monocytes in vitro express an active low density lipoprotein (LDL) receptor and an active receptor for negatively charged proteins, the scavenger receptor. When less than 15% of the lysine residues of human LDL were modified by malondialdehyde while the lipoprotein was in solution, recognition and uptake of the modified lipoprotein occurred via the LDL receptor. Further modification resulted in threshold recognition and uptake by the scavenger receptor with concomitant loss of recognition by the LDL receptor. The rate of degradation via the LDL receptor pathway was inversely related to the degree of modification whereas that mediated by the scavenger receptor was independent of the extent of incorporation of malondialdehyde once threshold recognition was achieved. In contrast to the interaction of LDL with malondialdehyde in solution, modification of less than 15% of the lysine residues of LDL adsorbed to heparin-Sepharose resulted in recognition and uptake by the scavenger receptor. The scavenger receptor-mediated uptake of malondialdehyde-modified LDL may be dependent on formation of recognition sites involving specific modified lysine residues or changes in the conformation of LDL induced by neutralization of specific lysine residues of the apoB polypeptides or both. PMID- 6281782 TI - The fate of 8-methoxypsoralen photoinduced crosslinks in nuclear and mitochondrial yeast DNA: comparison of wild-type and repair-deficient strains. AB - In Saccharomyces cerevisiae, after 8-methoxypsoralen [8-(OMe)Ps] photoaddition, more crosslinks are induced per unit dose in mitochondrial DNA than in nuclear DNA. In wild-type cells treated in the exponential phase of growth, single- and double-strand breaks are produced during crosslink removal and then are rejoined upon postexposure incubation. The incision step is almost blocked in the rad 3-2 mutant, which is also defective in excision-repair of UV-induced (254 nm) pyrimidine dimers. The cutting of crosslinks from nuclear DNA is depressed in wild-type stationary-phase cells. This is correlated with a higher sensitivity of such cells to 8-(OMe)Ps photoinduced cell killing. The incision of crosslinks is dramatically reduced in mitochondrial DNA. The rejoining of single- and double strand breaks is not only dependent on the product of the RAD51 gene (as shown by others) but also of the PSO2 gene. A correlation was found between the ability to recombine and strand rejoining. Therefore, as in bacteria, both the excision and the recombinational repair systems are involved in crosslink repair in yeast. However, double-strand breaks in yeast constitute repair intermediates which are not detected in Escherichia coli. The LD37 (dose necessary to induce a mean of one lethal hit per cell) corresponds to about 120 crosslinks per genome in exponential-phase cells of the wild type and to 1-2 crosslinks in the pso2-1 mutant. PMID- 6281783 TI - Compartmentalization of corticotropin-dependent steroidogenic factors in adrenal cortex: evidence for a post-translational cascade in stimulation of the cholesterol side-chain split. AB - A sensitive cell-free assay was developed for the analysis of corticotropin dependent factors that stimulate the rate-limiting step of adrenal steroidogenesis. In this assay adrenal post-mitochondrial supernates from corticotropin-stimulated rats caused a 10- to 100-fold increase in the de novo synthesis of pregnenolone and progesterone. A similar stimulation was observed by corresponding fractions from Leydig cells and mouse Y-1 adrenal tumor cells, but not from rat liver. Subcellular fractionation of rat adrenal tissue showed several steroidogenic factors to be present in various compartments. Recombination of them produced highly synergistic effects. The activation of some components could also be demonstrated in vitro, suggesting a cascade of events possibly linking the cAMP-dependent phosphorylation pathway with the rate limiting step. Cycloheximide prevented the production of these steroidogenic factors in vivo upon stimulation but had no effect in vitro, suggesting a post translational cascade involved in the activation of the cholesterol side-chain split. PMID- 6281784 TI - Photoaffinity-labeling and fluorescence-distribution studies of gonadotropin releasing hormone receptors in ovarian granulosa cells. AB - Photoaffinity labeling of rat ovarian granulosa cells and membrane preparations with a bioactive photoaffinity derivative of gonadotropin-releasing hormone resulted in identification of two specific components with apparent molecular weights of 60,000 and 54,000. Fluorescent visualization of gonadotropin-releasing hormone receptors in these cells, by using a bioactive rhodamine derivative of the hormone, indicated that the fluorescently labeled receptors were initially distributed uniformly on the cell surface and then formed patches that subsequently internalized (at 37 degrees C) into endocytic vesicles. These processes were dependent on specific binding sites for the rhodamine-labeled peptide on the granulosa cells. These studies may provide an experimental basis for understanding the molecular events involved in the action of the hormone in the ovary. PMID- 6281785 TI - [half-Cys4,half-Cys10]-alpha-Melanocyte-stimulating hormone: a cyclic alpha melanotropin exhibiting superagonist biological activity. AB - alpha-Melanocyte-stimulating hormone (alpha-melanotropin; alpha-MSH) is a linear tridecapeptide (Ac-Ser-Tyr-Ser-Met-Glu-His-Phe-Arg-Trp-Gly-Lys-Pro-Val-NH2) that reversibly darkens amphibian skins by stimulating melanomsome (pigment granule) dispersion within melanophores. By using a number of in vitro melanocyte assays, we have examined the conformational requirements for alpha-MSH activity. Synthesis of [half-Cys4,half-Cys10]-alpha-MSH, a cyclic, conformationally restricted, "isosteric" analogue of alpha-MSH, provided a melanotropin with a potency greater than 10,000 times that of the native hormone in stimulating frog (Rana pipiens) skin darkening. The cyclic analogue also showed substantially prolonged activity relative to the native hormone. [half-Cys4,half-Cys10]-alpha MSH was approximately 30 times more potent than alpha-MSH in stimulating lizard (Anolis carolinensis) skin melanophores in vitro. By using a cell-free Cloudman S 91 mouse melanoma plasma membrane preparation, we found the cyclic analogue to be approximately 3 times as potent as the native hormone in stimulating adenylate cyclase activity. These results provide insight into the conformational requirements for biological activity of alpha-MSH, and the comparative conformational requirements of alpha-MSH at a number of pigment cell receptors. PMID- 6281786 TI - Organization of human histone genes. AB - We describe the isolation and initial characterization of seven independent lambda Charon 4A recombinant phages which contain human histone genomic sequences (designated lambda HHG). Restriction maps of these clones and localization of the genes coding for histones H2A, H2B, H3, and H4 are presented. The presence of histone encoding regions in the lambda HHG clones was demonstrated by several independent criteria including hybridization with specific DNA probes, hybrid selection/in vitro translation, and hybridization of lambda HHG DNAs to reserve Southern blots containing cytoplasmic RNAs from G1-, S-, and arabinofuranosylcytosine (cytosine arabinoside)-treated S-phase cells. In addition, the lambda HHG DNAs were shown to protect in vivo labeled H4 mRNAs from S1 nuclease digestion. Based on the analysis of the lambda HHG clones, human histone genes appear to be clustered in the genome. However, gene clusters do not seem to be present in identical tandem repeats. The lambda HHG clones described in this report fall into at least three distinct types of arrangement. One of these arrangements contains two coding regions for each of the histones H3 and H4. The arrangement of histone genes in the human genome, therefore, appears to be different from that in the sea urchin and Drosophila genomes in which each of the five histone-encoding regions (H1, H2A, H2B, H3, and H4) is present only once in each tandemly repeated cluster. At least one clone, lambda HHG 41, contains, in addition to the histone genes, a region that hybridizes with a cytoplasmic RNA approximately 330 nucleotides in length. This RNA is not similar in size to known histone-encoding RNAs and is present in the cytoplasm of HeLa cells predominantly in the G1 phase of the cell cycle. PMID- 6281787 TI - Nucleosome segregation at a defined mammalian chromosomal site. AB - When animal cells replicate chromatin under conditions precluding new histone biosynthesis, half of the daughter DNAs are devoid of nucleosomes and are sensitive to staphylococcal nuclease. DNA sequences resistant to nuclease are associated with preexisting nucleosomes, which redistribute to progeny DNA duplexes during replication. We labeled newly replicated DNA sequences in a simian virus 40 (SV40)-transformed Chinese hamster cell clone with 5 bromodeoxyuridine (BrdUrd) in the presence and absence of a protein biosynthesis inhibitor, emetine. We resolved single-stranded BrdUrd- and dT-DNA sequences protected from nuclease digestion by nucleosomes and determined from which strands of the integrated viral DNA parental template (dT) and newly replicated progeny (BrdUrd) sequences were derived. Because we knew that the cell clone studied contained all of its integrated SV40 DNA at a single chromosomal site, we were able to determine that preexisting nucleosomes segregated to only one of the two daughter duplexes containing the integrated viral sequence. Additionally, in the presence of emetine, the integrated viral origin of replication, ORIsv, appeared not to function as a chromosomal replication origin, perhaps reflecting the drug's effect on synthesis of SV40 large tumor antigen. PMID- 6281788 TI - Conformational dynamics of a biologically active three-fragment complex of horse cytochrome c. AB - The conformational dynamics of a biologically active noncovalent complex containing three fragments, ferroheme fragment (1-25)H and apofragments (28-38) and [3H](56-104) [or [3H](39-104)], of horse cytochrome c has been studied with respect to kinetics and thermodynamics of dissociation. The rate of unfolding of the two-fragment complex ferro(1-25)H . (56-104) was also estimated. The results indicate that the ferrous three-fragment complex exhibits a higher frequency of dissociation-association with fragment (28-38) and a lower frequency of overall unfolding-folding at pH 7.0. In the presence of an excess of free (28-38) and below 30 degrees C, unfolding of the ferrous three-fragment complex appears to occur by activation to the transitional state without a large change in conformation, followed by virtually simultaneous dissociation of all three of the fragments [without going through the complex (1-25)H . (56-104), which is a major intermediate for folding]. Above 30 degrees C unfolding via the complex (1-25)H . (56-104) becomes detectable because the equilibrium between the two- and the three-fragment complex is highly temperature dependent. Thus, the relative probabilities of these two different ways of transition for unfolding are modulated by temperature. The observations suggest that the mode of activation of protein and hence the pathway for unfolding may vary depending on temperature. It is also suggested that the interatomic interactions binding the three fragments together in the ordered complex are linked to strengthen each other in the ground state. PMID- 6281789 TI - Structure of the insulin-like growth factor receptor in chicken embryo fibroblasts. AB - The insulin-like growth factors (IGFs) and insulin stimulate DNA synthesis and cell multiplication in chicken embryo fibroblasts in culture. This response appears to be mediated by interaction with a single type of IGF receptor. The present study examines the subunit structure of this receptor by covalently crosslinking two 125I-labeled IGFs, IGF-I and multiplication-stimulating activity (MSA), to chicken embryo fibroblasts by using disuccinimidyl suberate. After solubilization, NaDodSO4/polyacrylamide gel electrophoresis, and autoradiography, IGF receptor complexes of appropriate specificity were identified; they had Mr approximately 130,000 (major band) and approximately 260,000 (minor band) under reducing conditions and Mr greater than 300,000 without disulfide reduction. The proportion of the Mr 260,000 component increased with increasing concentration of crosslinking agent, suggesting that it was formed from smaller proteins during the crosslinking procedure. The IGF receptor in chicken embryo fibroblasts resembles the insulin receptor in size and structure but can be distinguished by a higher affinity for IGF-I and MSA than for insulin. Although IGF receptors with different structure and specificity have been recognized in other tissues, the function of these binding sites is unknown. The present study demonstrates that the IGF receptor of chicken embryo fibroblasts that appears to mediate the growth promoting effects of the IGFs contains a Mr approximately 130,000 binding subunit and exists as a native receptor complex of Mr greater than 300,000. PMID- 6281790 TI - Cell surface insertion of exogenous epidermal growth factor receptors into receptor- mutant cells: demonstration of insertion in the absence of added fusogenic agents. AB - We show that epidermal growth factor (EGF) receptor can be transferred in a biologically active orientation from donor hepatic membranes to recipient receptorless fibroblast cells. The recipient cells (NR-6) normally lack EGF receptors and are biologically unresponsive to EGF. The transfer of receptors from donor plasma membranes to recipient NR-6 surface membranes occurs in the absence of any added fusogenic agent. Studies on time and temperature dependence of this transfer indicate that it is due to preferential insertion of the EGF receptor over the other hepatic proteins. The inserted receptor is exceptionally stable to dissociation or damage, and this facilitated studies on its biological properties. The inserted receptor confers upon the hitherto unresponsive variant NR-6 cells a specific biological responsiveness to EGF as measured by EGF-induced stimulation of DNA replication and cell division. These findings suggest the existence of an affinity-mediated mechanism for the biologically active insertion of exogenous EGF receptors into receptorless variant cells. This insertion approach may be of use in the identification of receptor-associated membrane proteins that play a role in the transmission of EGF biological message. PMID- 6281791 TI - Inhibition of amiloride-sensitive sodium conductance by indoleamines. AB - To examine a possible role of indoleamines in the regulation of epithelial sodium absorption, the effect of serotonin (5-hydroxytryptamine) and several derivatives on electrolyte transport was measured in vitro in the baboon bronchus and in the trachea and colon of sodium-deficient rats. Serotonin, melatonin (N-acetyl-5 hydroxytryptamine), and harmaline (1-methyl-7-methoxy-3,4-dihydro-beta-carboline) inhibited sodium transport in all three preparations in a similar manner to the natriuretic agent amiloride. In all three epithelia, sodium absorption via the amiloride-sensitive pathway constitutes a substantial portion of total electrolyte transport, measured as the amiloride-sensitive short-circuit current. Thus 25 microM amiloride inhibited the short-circuit current 21% in the rat trachea, 63% in the baboon bronchus, and 90% in the rat colon. Serotonin, melatonin, and harmaline inhibited the amiloride-sensitive portion of the short circuit current from the luminal side of the epithelium. The inhibition was rapid, requiring only seconds, and maximal inhibition by serotonin was identical to that by amiloride. When sodium was omitted from the luminal solution, the short-circuit current was reduced a similar amount, suggesting that sodium absorption was being inhibited by both amiloride and the indoles. The IC50 value for amiloride was 50 nM in the baboon bronchus and 500 nM in the rat colon. In contrast, the IC50 value for serotonin was 0.4 mM in the baboon bronchus and 8 mM in the rat colon. These results, together with the wide distribution of amine precursor-uptake-and-decarboxylation (APUD) cells in the respiratory and intestinal tract, suggest that certain indoleamines could play a role as local regulators of fluid and electrolyte transport. For example, in the airways, indoleamines may be one of the factors involved in regulation of the depth of the periciliary fluid layer. PMID- 6281792 TI - Parathyroid hormone receptors in avian bone cells. AB - We have demonstrated binding of synthetic bovine parathyroid hormone (1-34) [bPTH (1-34)] to embryonic avian bone cells in monolayer culture. The binding sites have qualitative and quantitative characteristics of a physiologically important parathyroid hormone (PTH) receptor. At apparent steady state (60 min at 24 degrees C), 5-10% of electrolytically labeled, receptor-purified 125I-labeled bPTH-(1-34) bound specifically to the cells whereas nonspecific binding was less than 1% of the added labeled hormone. Scatchard analysis showed a single order of PTH binding sites (Kd = 0.6 nM) with approximately 10,000 sites per cell. In this bone cell system, PTH bound to its binding site and stimulated cAMP accumulation over the same concentration range. Bovine PTH-(1-84) bound to the cells with the same apparent affinity as bPTH-(1-34). PMID- 6281793 TI - Nonselective expression of simian virus 40 large tumor antigen fragments in mouse cells. AB - To understand the role of various functional domains of simian virus 40 early tumor antigens, we have cloned and introduced into mouse cells portions of early simian virus 40 DNA. Two types of truncated large tumor antigen (33 and 12.3 kilodaltons), as well as small tumor antigen, were identified by immunoprecipitation. Both truncated large tumor antigens have been found to be overproduced with respect to the small tumor antigen, although the 12.3 kilodalton truncated large tumor antigen was more stable than the 33-kilodalton one. Nonviral 53-kilodalton protein was not found associated with either truncated large tumor antigen in immunoprecipitations. PMID- 6281794 TI - gamma and alpha chains of human fibrinogen possess sites reactive with human platelet receptors. AB - Fibrinogen, a clottable plasma protein, agglutinates both prokaryotic cells (e.g., staphylococci) and eukaryotic cell fragments (e.g., platelets) through interaction with specific receptors. To identify the region of the fibrinogen molecule responsible for its interaction with human platelets, we prepared polypeptide chain subunits (alpha, beta, and gamma) of human fibrinogen by reduction and carboxymethylation. A mixture of the chains induced aggregation (clumping) of human platelets separated from plasma proteins and treated with ADP. When individual chains of fibrinogen were tested, gamma-chain multimers caused platelet aggregation at a molar concentration comparable with that of intact human fibrinogen. The beta chain remained inactive, and the alpha chain was 1/4th to 1/5th as reactive as the gamma chain. Monospecific antibody fragments against the gamma chain inhibited binding of 125I-labeled fibrinogen to the human platelet receptor and blocked aggregation of platelets induced by ADP in the presence of fibrinogen or gamma-chain multimers. These results indicate that the gamma chain of human fibrinogen bears the main site for interaction with the platelet receptor. PMID- 6281795 TI - Cloning of adeno-associated virus into pBR322: rescue of intact virus from the recombinant plasmid in human cells. AB - We have cloned intact duplex adeno-associated virus (AAV) DNA into the bacterial plasmid pBR322. The AAV genome could be rescued from the recombinant plasmid by transfection of the plasmid DNA into human cells with adenovirus 5 as helper. The efficiency of rescue from the plasmid was sufficiently high to produce yields of AAV DNA comparable to those observed after transfection with equal amounts of purified virion DNA. Thus, the recombinant plasmid itself may be a model for studying the rescue of a latent AAV viral infection. In addition, the efficient rescue of viable AAV from the recombinant plasmid should facilitate the genetic analysis of AAV. Finally, the results of an analysis of the DNA from rescued virions indicate that an inversion of the AAV terminal sequences occurred during replication. PMID- 6281796 TI - Batrachotoxin modifies the gating kinetics of sodium channels in internally perfused neuroblastoma cells. AB - We have studied the effects of batrachotoxin (BTX) on sodium channels in hybrid mouse neuroblastoma cells NG108-15 by using the suction pipet voltage clamp method. BTX-modified sodium channels activate with first-order kinetics and, over most of the potential range, activate more slowly than normal sodium channels. The peak conductance-voltage curve and the time constant of activation-versus voltage curve for BTX-modified sodium channels are shifted about 50 mV in the hyperpolarizing direction compared to the corresponding curves for normal sodium channels. There is no change in the slope of the conductance-voltage curve. These results suggest that BTX slows down one of the steps leading to channel opening, which consequently becomes rate-limiting. In addition, BTX eliminates both fast and slow inactivation. PMID- 6281797 TI - Progress toward a live, attenuated human hepatitis A vaccine. PMID- 6281798 TI - Membrane potentials, resistances, and conductances of toad bladder during Na+ - H+ transport and H+ transport. PMID- 6281799 TI - Essential fatty acids and adrenal steroidogenesis. PMID- 6281800 TI - The influence of ambient peroxides on the conversion of 5,8,11,14,17 eicosapentaenoic acid to prostaglandins. PMID- 6281801 TI - Early effects of EFA deficiency on the structure and enzymatic activity of rat liver microsomes. PMID- 6281802 TI - Metabolism of radioactive 5,8,11,14,17-eicosapentaenoic acid by human platelets. PMID- 6281803 TI - Effects of dietary alpha-linolenic acid on the production of a PGI-like substance of the isolated pulsatingly perfused rat aorta. PMID- 6281804 TI - Effects of high pressure on the interrelation between adrenergic neurotransmission and prostaglandins (PG) in the myocardium. PMID- 6281805 TI - Prostaglandins, essential fatty acids and cell-tissue interactions in immune inflammation. PMID- 6281806 TI - alpha-Linolenic and linoleic acids and the immune response. PMID- 6281807 TI - Cyclic AMP and prostaglandin levels in circulating mononuclear cells of cancer patients. PMID- 6281809 TI - Prostaglandins and cyclic-AMP in human semen. AB - Normospermic, oligospermic and azoospermic semen samples were analysed for prostaglandins of the E groups and for cyclic AMP. The whole procedure was rapidly performed in a nitrogen atmosphere. Higher levels of total PGEs (PGEs+19 hydroxy PGEs) were found in normospermic (135.6 micrograms/ml) than in either oligospermic or azoospermic semen. The average level of cyclic AMP in normospermic semen (32.0 nmole/ml) did not differ significantly from the test groups, but very low levels were found only in oligo and azoospermic semen. The correlation coefficient between cyclic AMP and total-PGE concentrations was +0.36. The correlation coefficient between cyclic AMP level and percentage of motile sperm cells was +0.56 (p less than 0.0005). Patients who were treated for two months with a daily dose of 50mg clomiphene citrate exhibited significant elevation of cyclic AMP in their seminal fluid. This finding suggests that cyclic AMP is important for sperm motility and that the clomiphene effect on human semen may be mediated by cyclic AMP. PMID- 6281808 TI - Deacylation of cellular lipids and arachidonic acid metabolism. PMID- 6281810 TI - Differential effect of prostaglandins and other products of arachidonic acid metabolism on measles virus replication in Vero cells. AB - The influence of cyclic adenosine 3', 5'-monophosphate (cAMP) and prostaglandin (PG), E1, E2, F2 alpha, A2 and thromboxane B2 (TxB2) and measles virus infection was investigated. Addition of PGE1, E2 and cAMP (10(-3)-10(-)8M) inhibited measles virus replication in Vero cells. TxB2 and PGA2 enhanced replication. Cytotoxic effects were not observed. Inhibition of infectious titers (98%) was most pronounced when agents were present throughout replication. Treated cells exhibited hemadsorbing antigen, but the size and number of cytoplasmic inclusions was decreased by cAMP. Pre-treatment of cells with PGE1 or presence of PGE1 during virus adsorption enhanced infectious titers (30%). Results suggest that cAMP and PGE1 inhibit measles virus production infection. That both enhancing and inhibiting effects of PGs were seen suggest that measles virus may require PG at specific times during the replicative cycle. PMID- 6281811 TI - Evidence for a mediator role of thromboxane A2 in the myotropic action of leukotriene B4 (LTB4) on the guinea-pig lung. AB - The mechanism of action of LTB4 has been investigated on the guinea-pig lung parenchymal strip. Mepacrine (20 microgram/ml), an inhibitor of phospholipase A2, abolished the action of LTB4 on parenchymal strips. Eicosatetraynoic acid (10 microgram/ml) and BW755C (40 microgram/ml) which are inhibitors of cyclooxygenase and lipoxygenase pathways, produced a marked inhibition of the lung strip contraction to LTB4. Similarly, aspirin (30 micrograms/ml) and flufenamate (1 microgram/ml) showed a strong inhibition of the contraction of parenchymal strips to LTB4; these results suggested that cyclooxygenase products mediate the action of LTB4. The response to LTB4 was unaffected by 15-hydroperoxyeicosatatraenoic acid (15-HPETE; 1 microgram/ml) while L8027 (25 ng/ml) reduced the contraction by 50%, suggesting that thromboxane A2 rather than prostacyclin was involved. Since parenchymal strips do not appear to be very sensitive to PGF2 alpha, PGE2 and the endoperoxides, and since effluents from LTB4-treated lungs produced contractions of lung strip and rabbit aorta which were reduced after 5 min. at 25 degrees, thromboxane A2 was postulated to mediate the lung effect of LTB4. The release of thromboxane B2 (TxB2) from lungs stimulated with LTB4 was confirmed by gas chromatography-mass spectrometric (GC-MS) analyses. PMID- 6281812 TI - pH dependent inhibition of serum oxytocinase activity by prostaglandins and cyclic GMP. AB - The effect of pH in the range 6.2 to 7.7 on the inhibition of serum oxytocinase (EC 3.4.11.3) activity by various compounds was studied using S-benzyl-L-cysteine p-nitroanilide (BCN) as substrate. Prostaglandins E1, E2, F2 alpha, 8-bromo-cGMP, cGMP, indomethacin and polyphloretin phosphate (PPP) produced a dose related pH dependent inhibition of serum oxytocinase. Their effect was maximum at pH 6.2 and minimum at pH 7.7. Hypertonic urea and saline also caused pH dependent inhibition; saline being most active at pH 6.2, and urea at pH 7.7. A similar pH dependent inhibition was found when these compounds were examined for their effect on the hydrolysis of L-leucine-p-nitroanilide (LN) by serum aminopeptidases at pH between 6.2 and 7.7. Although the inhibitors were more effective against the hydrolysis of LN than BCN substrate, cGMP and its 8-bromo derivative were more active against the BCN hydrolysis. cAMP, 8-bromo-cAMP, dibutyryl (db)-cAMP, db-cGMP, AMP, ADP, ATP, GDP, GTP, aspirin, sodium salicylate, paracetamol, theophylline and isobutylmethylxanthine (IBMX) at comparable concentrations and within the same pH range had no effect on the hydrolysis of either substrate. It is concluded that in serum obtained during pregnancy, the hydrolysis of LN may largely be attributed to oxytocinase activity. Thus, inhibition of LN hydrolysis by prostaglandins and other substances may be regarded as inhibition of oxytocinase activity. PMID- 6281813 TI - Characteristics of type 1 and type 2 benzodiazepine receptors in the ovine brain. AB - Analysis of the displacement of 3H-diazepam binding to membranes prepared from the ovine frontal cortex by the triazolopyradiazine CL218,872 yielded a Hill coefficient significantly below unity. By analogy with similar studies of this drug in rat brain this suggested the existence of Types 1 and 2 benzodiazepine receptors. The degree of displacement of 3H-diazepam by CL218,872 (200 nM, Type 1; 800 nM, Type 2) in homogenates of brain regions differed, the rank order being cerebellum greater than parietal cortex greater than frontal cortex congruent to temporal cortex congruent to hippocampus greater than striatum. Displacement of 3H-diazepam by CL218,872 was enhanced by 10(-5) M GABA in the striatum (at 200 nM and 800 nM CL218,872) and cerebellum (at 200 nM CL218,872). Benzodiazepine receptors in the ovine frontal cortex were least sensitive to CL218,872 (200 nM) in young fetuses (54-68 days gestation) and achieved adult levels of sensitivity by late gestation. Finally, the potency of CL218,872 to displace 3H-diazepam was not effected by the 3H-ligand concentration (0.5 nM or 5.0 nM), suggesting that Types 1 and 2 benzodiazepine receptors are not identical to the high and low affinity 3H-diazepam binding sites we have previously identified in the ovine brain. PMID- 6281814 TI - Potency and duration of action of the ACTH 4-9 analog (ORG 2766) as compared to ACTH 4-10 and [D-Phe7] ACTH 4-10 on active and passive avoidance behavior of rats. AB - Experiments were performed to examine the potency and duration of action of various ACTH analogs on active and passive avoidance behavior of rats. ACTH 4-10 and the ACTH 4-9 analog (ORG 2766) delayed extinction of pole-jumping avoidance behavior and facilitated passive avoidance responding. [D-Phe7] ACTH 4-10 facilitated extinction of pole-jumping avoidance behavior and facilitated passive avoidance responding. [D-Phe7] ACTH 4-10 facilitated extinction of pole-jumping avoidance behavior and facilitated passive avoidance responding. ORG 2766 was a thousand times more active than ACTH 4-10. The effect of ORG 2766 on extinction of pole-jumping avoidance behavior and on passive avoidance behavior was of longer duration than that of ACTH 4-10. As determined more precisely in the passive avoidance test it appeared that the action of ACTH 4-10 lasted 3 to 6 hours, while that of ORG 2766 amounted to at least 24 hours. Although [D-Phe7] ACTH 4-10 was a thousand times less active than ORG 2766 in the passive avoidance paradigm, its duration of action was of the same magnitude. In view of this, the marked increase in potency of the ACTH 4-9 analog cannot be explained only on the basis of its metabolic stability but also by an increased intrinsic activity. PMID- 6281815 TI - Induction of wet dog shakes by intracerebroventricular bethanechol in rats. Antagonism by neurotransmitter receptor blockers. AB - Bethanechol chloride, administered intracerebroventricularly, induces a characteristic wet dog shake (WDS) response in rats in a dose-related manner. WDS induced by bethanechol at the dose of 100 micrograms was antagonized by atropine, scopolamine and spiperone (muscarinic cholinergic and dopaminergic antagonists, respectively), whilst metergoline, methysergide, phentolamine, propranolol and bicuculline (serotonergic, alpha-adrenergic, beta-adrenergic and GABA-ergic antagonists) fail to inhibit this effect. The present experiments show that the shaking response may be produced by bethanechol, a potent muscarinic agent administered by the intracerebral route, and suggest that bethanechol-induced shaking behavior in rats may be a useful animal model for delineating agents with antimuscarinic activity. PMID- 6281816 TI - Effect of odor quality and intensity on conditioned odor aversion learning in the rat. AB - Odor quality and intensity were varied to test the ability of rats to associate odor with an induced illness. Rats were allowed 10 minutes access to water on each of nine days; deodorized air was directed towards each rat's nose while drinking at familiarization and recovery sessions (days 1-5 and 7-8, respectively) and odorized air at treatment and test sessions (days 6 and 9, respectively). Each rat was injected with LiCl following its drinking period on day 6. The difference between the amount of water consumed on day 6 and day 9 gave a measure of the conditioned aversion. Only mild or no aversion occurred with odors of n-butyric acid, benzylamine, cyclohexanone, and n-butanol. Strong conditioned aversions were obtained to odors of triethylamine, 1,4-cineole, and isoamyl acetate, and the degree of aversion increased linearly with the log of odor concentration. The effect of odor quality, intensity and presentation method, and the role of the different chemoreceptor systems in the acquisition of odor aversions are discussed. PMID- 6281817 TI - beta-Endorphin-induced psychomotor excitation in the cat. AB - The effect on behavior of synthetic human beta-Endorphin injected into the cerebral ventricles of the cat was investigated in these experiments. beta endorphin produced psychomotor excitation (i.e., restlessness, apprehension, flight and locomotion), accompanied by pupillary dilatation and tremor. Between periods of locomotion, the cat sat moving its head from side to side with eyes wide open and mydriasis, or stood stiffly with a vacant stare, pupils dilated and eyes wide open. During this time the cat did not react to objects moving in front of it. Behavioral changes produced by a single dose of beta-endorphin were dose dependent and long-lasting. Pretreatment with intracerebroventricular nalorphine depressed or abolished the behavioral changes, mydriasis and tremor caused by beta-endorphin. It is concluded that beta-endorphin acts on central opiate receptors promoting psychomotor excitation. PMID- 6281818 TI - Dental pain and injection of agents into the trigeminal nucleus of guinea pigs. AB - The effects of microinjections of carbachol (Carb), serotonin (5-HT) and noradrenaline (NOR) into the spinal trigeminal nucleus (STN) of guinea pigs were studied in superficially anesthetized animals (nembutal, 40 mg/kg) submitted to noxious dental stimulation (electric shock). The defense motor response (DMR) was studied during noxious stimulation through the control period and up to 60 minutes after chemical stimulation. The agents were microinjected into four sites in the bulbar-spinal projection of the STN. Carb (1 microgram/microliter) caused significantly decreased DMR when injected into all regions studied, with total DMR abolition into the site located 1.0 mm caudal to the obex. 5-HT (30 micrograms/microliters) and NOR (30 micrograms/microliters) elicited a less intense effect than Carb, causing significant reduction in DMR only when injected into two sites. These results suggest cholinergic, serotonergic and adrenergic involvement of the mechanism of aversive response to dental pain. PMID- 6281819 TI - In utero taste/odor aversion conditioning in the rat. AB - Rat fetuses exposed to a taste-odor stimulus and an aversive stimulus in utero showed an aversion to the taste/odor stimulus when tested 16 days postnatally. Fetuses which received various control treatments in utero did not show an aversion. These data show that rat fetuses at 20 days of gestation are capable of associative learning which can be demonstrated more than two weeks postnatally. PMID- 6281820 TI - Vagal unitary responses to intestinal amino acid infusions in the anesthetized cat: a putative signal for protein induced satiety. AB - Single unitary discharges in the nodose ganglia were recorded extracellularly in chloralose anesthetized cats while amino acid solutions were being perfused through the small intestine via implanted cannulae. Test infusions consisted of either amino acid mixtures (12 amino acids; 120 mM in all) or individual amino acids (50 mM each) dissolved in Krebs Henseleit buffer. Units which were activated by amino acid infusions were also tested with 10% glucose infusions performed in the same way. Control infusions consisted of either buffer alone or a physiological saline solution isotonic to the test solution. All perfusions were performed at 38 degrees C, pH 7.4 by means of a syringe over a 10 second period. Out of 1250 vagal units activated by electrical vagal stimulation, 92 units showed an increased firing rate in response to amino acid intestinal perfusions. Of these, only 1/7 were also responsive to glucose perfusions. Osmotic, thermal or mechanical stimuli associated with infusions did not modify vagal responses to the amino acids. Among vagal units responding only to amino acid but not to glucose infusion, some were activated in a specific manner, depending on the specific amino acid infused intraduodenally. These neurons illustrated a very strict specificity regarding the nature of chemical stimuli. The very short latency, mean of 9 sec +/- 0.7 (SE) of these vagal neurons to amino acid infusions unequivocally indicates that chemoreceptors are located at the preabsorptive level. The corresponding fibers were non-myelinated (conduction velocities: 0.8/1.4 m/sec.) and were of the C type. The functional characteristics of these vagal amino acid receptors are discussed in terms of the role of intestinal signals in short term protein satiety. PMID- 6281821 TI - Involvement of endogenous opioids with forced swimming-induced immobility in mice. AB - The present study investigated the involvement of endogenous opioid mechanisms with the immobility response induced in mice by forced swimming. Pretreatment with the narcotic antagonist naloxone (0.625--40.0 mg/kg) caused a dose-dependent decrease in the duration of immobility in mice subjected to a 10 min swim test. This effect was more pronounced in C57BL/6J mice than in BALB/C mice. A low dose of morphine (0.15 mg/kg) potentiated immobility whereas higher doses (0.625/10.0 mg/kg) had no demonstrable effect on immobility in these strains. The results suggest that release of endogenous opioids may be a physiological event promoting natural cataleptic-like behaviors in mice. PMID- 6281822 TI - Differential hormonal and physiological responses to stress in Roman high- and low-avoidance rats. PMID- 6281823 TI - Differential effects of medial, central and basolateral amygdaloid lesions on four models of experimentally-induced aggression in rats. AB - To clarify whether various nuclei of the amygdaloid complex play different roles in aggressive behavior including muricide, 4 types of aggression were experimentally induced in rats. These include olfactory bulbectomy (OB rats), midbrain raphe lesions (Raphe rats), administration of delta 9 tetrahydrocannabinol (THC rats) and long-term isolation (Iso rats). Rats which exhibited muricide following these treatments were subjected to bilateral lesions of either the medial (AME), central (ACE) or basolateral (ABL) amygdaloid nuclei. Both muricide and hyperemotionality in the OB rat were markedly inhibited by AME lesions. Those of the Iso and THC rats were moderately inhibited. However, in the Raphe rat, aggressive behavior was not inhibited by AME lesions. Furthermore, ACE or ABL lesions caused no significant changes in all 4 models of aggression. These results suggest that the AME plays a facilitatory role in aggression of OB, Iso and THC rats, but aggression in Raphe rat is independent of amygdaloid activity. PMID- 6281825 TI - The influence of ACTH fragments on habituation of the prey catching behaviour in the European toad (Bufo bufo L.). AB - Earlier observations on the influence of the ACTH molecule on habituation in toads revealed a capacity, which resembles the improvement of learning and memory processes in rats [6]. In the present investigation ACTH, ACTH1-10 and ACTH11-24 were tested on habituation of turning reaction, a stereotypic component of the prey catching behaviour in the European Toad, Bufo bufo L.. Furthermore the time relationship of the ACTH-effect was determined by injections at different times prior to the beginning of the habituation. ACTH as well as ACTH1-10 caused a dramatic decline in turning reactions per minute towards a prey dummy, when administered 30 min prior to habituation. Therefore, the capacity to facilitate behavioral adaptations must be located mainly in the first ten amino acids of ACTH. PMID- 6281824 TI - Opiate receptors, food intake and obesity. AB - The present studies tested the effect of acute and chronic administration of naloxone on food intake of lean and genetically obese (ob/ob) mice. Acute administration of naloxone, a drug which blocks opiate receptors, produced a greater reduction of food intake in obese (ob/ob) mice than in the lean littermates. For chronic experiments with naloxone, the daily feeding period was shortened to eight hours and two injections of naloxone were given four hours apart. With this procedure of scheduled-feeding the food intake of both lean and obese mice was depressed during the first hour after injecting naloxone. However, beginning on the second day of treatment, the lean mice began to eat more food than the untreated controls during the eight hour feeding period. Food consumption by lean mice reached values 140 to 200% above the control levels between the fourth and sixth day. In the obese mice the rise in food intake was more gradual and did not reach 200% of the control value until the sixth day. Body weight changes reflected the changes in food intake. In contrast to naloxone, chronic treatment with morphine lowered food intake and blocked the stimulatory effect of naloxone. Our findings suggest that endogenous opioids may play a role in signalling satiety and in regulating long-term energy balance. PMID- 6281826 TI - Endogenous opioids interact in stress-induced hyperglycemia in mice. AB - Intermittent inescapable foot shock stress for 1 hour elicited significant hyperglycemia in mice. Pretreatment with the long acting narcotic antagonist naltrexone (1.0 mg/kg, 1 hr prior to stress) prevented stress hyperglycemia. Naltrexone did not affect blood glucose in unstressed control mice. These findings suggest the involvement of endogenous opioids in the hyperglycemic response to stress in mice. The possible mode of interaction of endorphin in stress hyperglycemia is discussed. PMID- 6281827 TI - The suppression of sucrose intake by cholecystokinin is scaled according to the magnitude of the orosensory control over feeding. AB - The intestinal hormone cholecystokinin (CCK) has been shown to play a role in the termination of food intake, however its behavioral mechanism of action remains to be determined. Recent work from this laboratory suggested that the suppression of intake with CCK is dependent upon the specific orosensory characteristics of the substance being consumed and that the hormone may suppress intake by altering the behavior maintaining characteristics of orosensory stimuli. The present study further investigated this suggestion by determining whether changes in the orosensory characteristics of food after the magnitude of the suppressive effect of CCK. Specifically, the magnitude of the CCK effect on the intake of sucrose solutions of various concentrations was determined on non-deprived rats. The results of this work indicate that the suppressive effect of synthetic CCK (CCK 8) cannot be overridden by increases in sucrose concentration. In contrast, it was found that over a range of sucrose concentrations, the magnitude of the CCK effect increased with solution concentration. Because sucrose concentration predicts both caloric density and the magnitude of orosensory control (as measured by grams consumed), it appears that CCK may act on this control to regulate meal size in proportion to the caloric density of the food. PMID- 6281828 TI - Dual effect of response preparation on conditioned H reflex. AB - The late or intercurrent facilitation of the soleus H reflex recovery cycle was studied during the preparatory period of a visual reaction time task. Response preparation produced three kinds of effect on the conditioned reflex elicited 200 msec before the end of the preparatory period: depression of the conditioned reflex which was maximum at the 150 msec conditioning interval; enhancement of the conditioned reflex which was maximum at the 300 msec conditioning interval; and, when the soleus muscle was involved in the execution of the voluntary response, an additional depression of the conditioned reflex at each conditioning interval. These different effects of response preparation are discussed with reference to the neural mechanisms which are thought to mediate the intercurrent facilitation. PMID- 6281829 TI - Effects of stimulus and task factors on Achilles tendon reflexes evoked early during a preparatory period. AB - The effects of intensity, duration and modality of a warning signal on tendon (T) reflexes evoked during the initial phase of a preparatory period of 4 sec were investigated. Reflexes were evoked simultaneously in both legs, from 0 to 350 msec after warning signal onset in steps of 50 msec. The required response was a planter flexion of the right foot. A facilitation of reflexes was seen within 150 msec after warning signal onset, showing a somewhat longer latency for visual as compared to auditory signals. An effect of intensity was found in the auditory modality only, where the louder of two warning signals yielded a clear peak at 100 msec while the softer stimulus caused no significant departure of Achilles tendon reflexes from baseline. The time course of facilitation in the auditory modality was influenced by warning signal duration as well, although this effect was only marginally significant. There were no effects of physical warning signal parameters on reaction time. A comparison with an experiment in which non-signal stimuli were presented alone, pointed to aspects of the preparatory process which were manifest at the spinal level as early as 200 msec after warning signal onset. PMID- 6281830 TI - Conditioned aversion to a taste perceived while grooming. AB - Four experiments were conducted to determine how the characteristics of conditioned taste aversion (CTA), as described from studies conducted in the drinking and feeding contexts, applied in the grooming context. In Experiment 1, sodium saccharin was mixed with a "neutral-tasting" jelly and applied to the fur of male Sprague-Dawley rats. Rats injected with LiCl after the applications strongly avoided saccharin solutions in subsequent 1-hr, 2-choice (Saccharin solution vs water) drinking tests, whereas rats injected with NaCl or given plain jelly on the fur showed only an initial neophobic response to the saccharin solution. Thus, the taste of saccharin was perceived while grooming and the CTA formed in the grooming context generalized to drinking. In experiments 2-4, we obtained evidence that: (a) rats discriminated between one intensity of saccharin applied to the fur and another used in the test solution; and (b) rats differentiated between qualities of the two tastants applied to the fur in that saccharin overshadowed NaCl; and (c) taste qualities were more important than toxic properties when two stimuli (Saccharin, LiCl) were used (saccharin overshadowed NaCl in subsequent drinking tests). We speculate that taste, while grooming might play a role in social communication in some vertebrates. Further, CTA and grooming might have uses in rodent control (e.g., in agricultural situations) not previously considered such as in delivering a non-attractive, low salience toxin so that the taste of the crop overshadows that of the bait, and induces crop aversion. PMID- 6281831 TI - Construction and application of R prime plasmids, carrying different segments of an octopine Ti plasmid from Agrobacterium tumefaciens, for complementation of vir genes. PMID- 6281832 TI - A functional map of the replicator region of the octopine Ti plasmid. PMID- 6281834 TI - Incompatibility and host range of pGD10 from Capnocytophaga ochraceus, formerly Bacteroides ochraceus. PMID- 6281833 TI - Genetic studies of F plasmid maintenance genes involved in copy number control, incompatability, and partitioning. PMID- 6281835 TI - GABA and acute psychoses. PMID- 6281836 TI - Jumping behavior induced by thyrotropin releasing hormone in combination with apomorphine in mice. AB - Neuronal mechanisms involved in the jumping produced by thyrotropin releasing hormone (TRH) plus apomorphine were studied in mice. TRH (5--20 mg/kg IP) administered after apomorphine (0.25--2.0 mg/kg IP) elicited jumping behavior, the frequency of jumping being proportional to the dose of TRH but inversely related to the dose of apomorphine. Jumping behavior was also induced by TRH (20 mg/kg IP) administered after atropine (5 mg/kg IP) or clonidine (0.5 mg/kg IP). The jumping elicited by TRH (20 mg/kg IP) in combination with apomorphine (0.25 mg/kg IP) was decreased by pretreatment with haloperidol (1 mg/kg IP), physostigmine (0.2 mg/kg IP) or phentolamine (10 mg/kg IP), unaffected by propranolol (10 mg/kg IP), and markedly increased by atropine (5 mg/kg IP) or clonidine (0.5 mg/kg IP). The results suggest that TRH in combination with apomorphine, atropine or clonidine elicits jumping in which dopaminergic, cholinergic and noradrenergic functions are concomitantly involved. PMID- 6281837 TI - Anticholinergic activity of imipramine and some analogs at muscarinic receptors of cultured mouse neuroblastoma cells. AB - Imipramine and some of its analogs (trimipramine, 3-chlorimipramine, desipramine, 3-chloro-2-hydroxyimipramine, 2-hydroxyimipramine, and didesmethylimipramine), were assayed for their potencies as antimuscarinic agents by their abilities to antagonize muscarinic receptor-mediated cyclic guanosine monophosphate (GMP) formation by cultured mouse neuroblastoma cells. Equilibrium dissociation constants for these compounds yielded the following rank order of potency at the muscarinic receptor: imipramine greater than trimipramine greater than 3 chlorimipramine greater than desipramine greater than 3-chloro-2 hydroxyimipramine greater than 2-hydroxyimipramine greater than didesmethylimipramine. These results indicate that didesmethylation of the side chain nitrogen or hydroxylation of the ring at the 2-position lead to marked reductions (30-fold and 12-fold, respectively) in antimuscarinic activity of imipramine. PMID- 6281838 TI - THIP inhibits feeding behavior in fasted rats. AB - The effects of 4,5,6,7-tetrahydroisoxazolo-[5,4-c]pyridin-3-ol (THIP) were compared with those of d-amphetamine and GABA in fasted rats. Intravenously administered THIP produced a dose-dependent decrease in food consumption (ED50 congruent to 1.5 mg/kg) by an action that was not reversed by prior subcutaneous or simultaneous intravenous (IV) injection of bicuculline. d-Amphetamine-SO4 also produced a decrease in food consumption in this model (ED50 congruent to 0.2 mg/kg, IV). Unlike THIP, GABA (in doses up to 100 mg/kg, IV) did not produce a marked anorexigenic effect. These results provide further evidence that THIP can penetrate the "blood-brain barrier", and that central GABA-ergic systems are involved in controlling food intake. PMID- 6281839 TI - Repeated administration of subconvulsant doses of GABA antagonist drugs. I. Effect on seizure threshold (kindling). AB - Repeated subconvulsant doses of the GABA antagonist drugs picrotoxin (5 mg/kg), pentylenetetrazol (PTZ) (30 mg/kg), and bicuculline (3.5 mg/kg), were given IP once daily to rats. Picrotoxin produced rapid kindling to full seizures in about 5 days, PTZ produced sporadic myoclonic seizures after 17 days, PTZ produced sporadic myoclonic seizures after 17 days whereas bicuculline only produced occasional mild jerking. Following these treatments, seizure thresholds to these drugs were measured by an IV infusion method to minimise problems of systemic uptake and metabolism of the drugs. Picrotoxin- and PTZ-treated animals showed no alteration in seizure threshold. However, following bicuculline pretreatment, seizure threshold was raised. Methodological problems in the interpretation of pharmacological kindling are discussed. PMID- 6281840 TI - Interactions between narcotic agonists, partial agonists andd antagonists evaluated by punished an unpunished behavior in the rat. AB - The effects of morphine, ketocyclazocine, cyclazocine, and SKF-10,047 were tested alone and in conjunction with naltrexone or naloxone, in rats responding under a multiple fixed-interval 3-min schedule of food presentation. Under this paradigm, electric shock was delivered on a fixed-ratio schedule for responses occurring during alternate schedule components. All of the drugs (except naltrexone and naloxone) decreased average rates of responding maintained by the unpunished component in a dose-dependent manner. The rate-decreasing effects of morphine and ketocyclazocine were antagonized by naltrexone. The rate-decreasing effects of cyclazocine were only slightly reversed by the antagonists, while those effects of SKF-10,047 were not affected by naltrexone. In some animals, certain doses of SKF-10,047 increased unpunished responding. This rate-increasing effect was antagonized by naltrexone. Morphine, ketocyclazocine, cyclazocine, and SKF-10,047 increased responding that was suppressed by electric shock, and these increases were antagonized by naltrexone and naloxone. Thus, the antagonism of opiate effects by narcotic antagonists depends in part on the behavior being evaluated. PMID- 6281842 TI - Internalization of ouabain and replacement of sodium pumps in the plasma membranes of HeLa cells following block with cardiac glycosides. PMID- 6281843 TI - Sodium movements across the vascularly perfused anuran small intestine and colon. AB - Values of unidirectional Na fluxes measured across the vascularly perfused small intestine of three anuran species are higher than those found in other preparations in vitro. In Rana ridibunda and R. pipiens no net movement of Na across the small intestine can be detected. In contrast, the unidirectional fluxes of Na across the colon of R. ridibunda and R. pipiens are lower than across the small intestine and a significant net absorption is found. Apparent loading and unloading pools for Na within the small intestine, as measured with tracer Na under standard experimental conditions, consist largely of extracellular Na presumably within the bulk phase of the lumen. The size of these pools can be greatly reduced by the rapid addition to or removal from the lumen of tracer. The loading pool appears to occupy not more than about 9% of the total tissue water, equivalent to about 20% of the extracellular water of the tissue. The washout of tracer Na from preloaded small intestine into the vascular bed is bi-exponential and appears from a pool, or pools, of apparent greater size than that of the loading pool. The results show that Na can move very rapidly across the small intestine and suggest that a high proportion of this movement occurs possibly via paracellular shunt pathways. PMID- 6281841 TI - Dyphenylhydantoin enhancement of diazepam effects on locomotor activity in mice. AB - The interaction of diazepam and diphenylhydantoin on locomotor activity and rearing behavior was studied in mice. Pretreatment of mice with diphenylhydantoin (4.0 and 8.0 mg/kg) significantly reversed the stimulatory effects of low doses of diazepam and considerably increased the depressant effects of the benzodiazepine on locomotor activity and rearing. Neither diazepam (up to 4.0 mg/kg), diphenylhydantoin (8.0 mg/kg) alone, nor combined treatment with both drugs affected brain GABA level and glutamic acid decarboxylase (GAD) activity at any dosage used. The present behavioral and biochemical data suggests that some of the pharmacological effects of diazepam need not be related to GABAergic mechanisms. PMID- 6281844 TI - The nature of the atrial receptors responsible for a reflex increase in activity in efferent cardiac sympathetic nerves. AB - In dogs anaesthetized with chloralose, distension of small balloons in the right upper and middle pulmonary vein-atrial junctions, to stimulate left atrial receptors, caused an increase in heart rate and an increase in activity in efferent sympathetic nerve fibers in cardiac branches of the right stellate ganglion. Cooling of the cervical vagi in steps reduced the magnitude of the responses in these sympathetic nerve fibres. In four dogs, the response in six preparations of sympathetic nerves was slightly reduced with the vagi at 18 degrees C and markedly reduced or abolished at 12 degrees C. In these nerves there was no significant response to distension of the balloons when the cervical vagi were cooled to 9 degrees C. The effect of cooling the vagi was the same as the previously shown effect of cooling on the increase in activity in myelinated afferent vagal fibres and the increase in heart rate during stimulation of atrial receptors. It is concluded that the increase in activity in efferent sympathetic cardiac nerve fibres during distension of small balloons in the pulmonary vein atrial junctions involves receptors discharging into myelinated vagal nerve fibres; receptors which discharge into non-myelinated vagal nerves of afferent sympathetic nerve fibres are not involved in this response. Thus, the efferent sympathetic nerve fibres studied are likely to represent the efferent pathway of the response of an increase in heart rate to distension of the small balloons. PMID- 6281845 TI - The response in efferent cardiac sympathetic nerves to stimulation of atrial receptors, carotid sinus baroreceptors and carotid chemoreceptors. AB - The reflex increase in activity in efferent cardiac sympathetic nerve fibres in response to distension of small balloons in the pulmonary vein-atrial junctions was studied in dogs anaesthetized with chloralose; the carotid sinuses were isolated and perfused with blood. In the efferent cardiac nerves which responded to distension of the balloons there was no significant change in the control activity, or in the magnitude of the response during changes in carotid sinus pressure from 8 to 30 kPa, during changes in the blood perfusing the sinuses from arterial to venous or during stimulation of the cutaneous branch of the left radial nerve. In contrast, in efferent cardiac nerves which did not respond to the distension of the balloons, consistent and significant changes in activity resulted from stimulating arterial baroreceptors, chemoreceptors or a cutaneous branch of the radial nerve. It is concluded that efferent cardiac sympathetic nerve fibres which respond by an increase in activity to stimulation of the atrial receptors, constitute a group of nerve fibres which are separate from those responding to stimulation of receptors in the carotid region or to stimulation of peripheral somatic nerves. PMID- 6281846 TI - Temperature dependence of the sodium channel gating kinetics in the node of Ranvier. AB - Temperature dependence of the Na+ channel gating kinetics was measured from the ionic and charge displacement currents in the node of Ranvier of Xenopus laevis, m3h kinetics was applied, assuming a delay, delta t, in the activation process. The rate constants for the m- and h-process showed Arrhenius temperature dependence with Q10 of 2.34 and 2.9 respectively, while delta t exhibited non Arrhenius temperature-dependence. Q10 for PNa, measured as 1.6, was smaller than for the rate constants and similar to that for a diffusion process. A negative shift and decrease in voltage sensitivity of the steady-state curves, h infinity and m infinity, occurred with decreasing temperature. The maximum time constant obtained from a single exponential fit to the displacement currents during the pulse for times greater than 90 microseconds exhibited Q10 of 2.01, which lies between that for PNa and that for tau m. PMID- 6281847 TI - beta-Adrenergic effects on composition of parotid salivary secretion of sheep on feeding. AB - Observations were made in sheep, before and after fresh food was given during teasing with food and after rumination, on the flow of parotid saliva and its protein Mg2+, K+, Na+ and Cl- concentrations. The animals studied had either a cannulated or fistulated parotid salivary duct. Parotid salivary flow, protein, Mg2+, K+ and Cl- increased markedly following feeding. The increases in protein and Mg2+, but not in flow, were largely blocked by the i.v. administration of propranolol (1 mg . kg-1). Whereas the actual ingestion of food was associated with large increases in protein (up to 42.5 times, to as high as 1760 micrograms . ml-1 of saliva), teasing with food caused relatively minor increases in parotid saliva. There were slight, if any, changes in protein concentration during the increased parotid salivary flows of rumination, whether chewing was on the same side or contralateral to the cannulated parotid salivary duct. It is concluded that a beta-adrenergic mechanism previously demonstrated in acute experiments contributes to increases in the secretion of protein of the parotid saliva when sheep eat. There was a close correlation between the concentrations of protein and of Mg2+ but not of the other electrolytes studied. PMID- 6281848 TI - The dorsal horn of the spinal cord. AB - Recent advances in techniques, especially the intraneuronal injection of the enzyme horseradish peroxidase, have led to a new ear in our understanding of spinal cord structure and function. Input to the cord is precisely organized: the primary afferent fibres from different types of receptors distribute their anatomically specific collaterals to particular parts of the dorsal horn, afferent fibres from the skin lay down a precise somatotopic map, input to the dorsal horn from descending systems is also distributed in a localized way. The neurones of the dorsal horn are varied in both structure and function, even so some quite specific cell types can be identified and the dendritic trees may respect laminar boundaries as determined cytoarchitectonically (although the majority of neurones have dendrites that cut across these boundaries). The output pathways from the dorsal horn are many and various, but again they arise from cells in definite parts of the dorsal horn. The dorsal horn must be considered as a well-organized, and complex, part of the central nervous system. It cannot be considered as a structural or functional unit but is made up of many interacting parts that process input from the primary afferent fibres, from other levels of the spinal cord and from many descending pathways from the brain. PMID- 6281849 TI - [A rare case of particularly extensive odontodysplasia (I)]. PMID- 6281850 TI - Multifocal glioblastoma: diagnostic implications. PMID- 6281851 TI - 99mTc red blood cell venography in deep vein thrombosis of the leg: a correlation with contrast venography. PMID- 6281852 TI - [Role of higher order structure of DNA in initiation process of DNA replication (author's transl)]. PMID- 6281853 TI - [Operator hypothesis in presynaptic membrane and vesicular hypothesis in synaptic vesicle (author's transl)]. PMID- 6281854 TI - Effects of solution of low pH and taurocholate on release of beta-endorphin-like immunoreactivity from human duodenal mucosa in vitro. AB - The presence of beta-endorphin-like immunoreactivity (beta-EpLI) in human duodenum and its release were studied. beta-EpLI was detected in the duodenum (mucosa, 26.7 +/- 6.3 pmol/g wet weight, mean +/- SEM; remaining tissue 23.1 +/- 5.3 pmol/g wet weight) and the stomach (7.1 pmol/g wet weight). The two activities gave similar curves for inhibition of beta-Ep radioimmunoassay of synthetic beta-Ep. On gel-filtration chromatography of a duodenal extract, two components of beta-EpLI were separated. When human duodenal mucosa was perfused with a solution of pH2 or 1mM or 5mM taurocholate, the release of beta-EpLI from mucosa into the perfusate increased 2-4 fold. These results indicate that beta EpLI present in human duodenal is released by the direct action of low pH or taurocholate on the duodenal mucosa and suggest that it may have a physiological role. PMID- 6281855 TI - [Calcium and arterial hypertension]. PMID- 6281856 TI - [Splenohepatoplasty: is it an inhibitory surgical technic for hepatic cancer?]. PMID- 6281857 TI - [Immunofluorescence with anti-alpha-1-antitrypsin antibody in hepatocarcinoma]. PMID- 6281858 TI - [Isolated idiopathic hypoparathyroidism with late presentation. Comments on a case with unusual presentation]. PMID- 6281859 TI - [Epidemiology and clinical incidence of bovine trypanosomiasis in the North of the Ivory Coast]. PMID- 6281860 TI - [Economic evaluation of losses incurred by trypanosomiasis in 4 genetic types of cattle in the North of the Ivory Coast]. PMID- 6281861 TI - [Effects of alpha-ketoglutarate of imidazole on fentanyl-induced bradycardia bradydypnoea syndrome. Experimental study (author's transl)]. PMID- 6281862 TI - [Current findings on the physiopathogenesis and diagnosis of diarrhea]. PMID- 6281863 TI - Interactions of starvation and selective phosphorus depletion on renal phosphate reabsorption. AB - Renal phosphate (Pi) wastage following 7 days of starvation was investigated in normal rats (HI-P) and others previously stabilized on a low phosphorus (LO P)diet. In LO-P animals, Pi excretion increased after starvation, but was significantly less than in starved HI-P rats. After thyroparathyroidectomy, the increase in Pi excretion after parathyroid hormone (PTH) was significantly greater in nonacidotic starved HI-P rats than in LO-P animals. However, PTH elicited a 31-fold increase in Pi excretion in both of these groups. Starved LO-P and HI-P rats responded equivalently to dibutyryl cyclic AMP. The renal response to phosphate depletion normally promotes Pi conservation, but is attenuated markedly by 7 days of subsequent starvation. This results from at least partial restoration of phosphaturic responsiveness to PTH during starvation. PMID- 6281864 TI - Temperature dependence if beta-receptor mediated relaxation in rabbit intestine. AB - The relaxation induced by isoproterenol, A beta-stimulant, appears more markedly at 25 degrees C (pD2 7.98) than at 37 degree C (pD2 6.88) in rabbit intestine. The isoproterenol-induced relaxation was antagonized by treatment with propranolol but not phentolamine at both 25 degree and 37 degrees C, indicating that the relaxation was mediated via adrenergic beta-receptor. The relaxations induced by papaverine, theophylline and MgCl2 were not influenced by temperature change. Basal tissue cyclic AMP content was higher at 25 degrees C than at 37 degrees C, and isoproterenol increased the tissue cyclic AMP content to a similar extent at 25 degrees C and 37 degrees C in the presence of theophylline. The binding of 3H-dihydroalprenolol (3H-DHA) to beta-receptor did not solve the reason of higher sensitivity to isoproterenol at 25 degrees C than at 27 degrees C. At 37 degrees C isoproterenol was more effective in producing a relaxation in the presence than the absence of ascorbic acid (0.057 mM). It is thought that the temperature dependency of the isoproterenol-induced relaxation may be mainly due to the breakdown of isoproterenol in a high temperature. PMID- 6281865 TI - Effects of dithiocarb and (+)-catechin on microsomal enzyme activities of rat liver. AB - The effects of dithiocarb and (+)-catechin on the microsomal mixed-function oxidase system of rat liver were investigated after a single dose as well as after repeated treatment for 7 and for 28 days. Dithiocarb (200 mg/kg p.o.) significantly reduced the microsomal cytochrome P-450 content, aniline hydroxylase and aminopyrine demethylase activities; maximum decrease was observed at 4 hrs, return to normal values after 24 hrs. (+)-catechin (200 mg/kg p.o.) had no effects in this respect. Both agents did not affect microsomal enzyme activities when applied orally for 7 days. After 28 days treatment only dithiocarb (50 - 100 - 200 mg/kg p.o.) exerted a dose-dependent depression of the aniline hydroxylase activity. In vitro experiments confirmed the in vivo observations: a concentration-dependent inhibition of the aniline hydroxylation and aminopyrine demethylation were seen from the addition of dithiocarb, the I50 values were 5.4 X 10(-6) M and 9.6 X 10(-5) M, respectively. (+)-catechin showed no inhibitory activity on both enzyme activities in vitro. Both dithiocarb and (+)-catechin depressed the activity of the NADPH-cytochrome C-reductase only in the 10(-4) M concentration range, these effects should be therefore evaluated as non-physiological without relevance in vivo. PMID- 6281866 TI - Addition of fiber (wheat bran, bassorin) to the standard food does not influence biliary lipid composition in piglets. AB - The influence of an additional fiber diet on biliary lipid composition of gallbladder bile was studied in 12 piglets. In addition to the standard feed wheat bran was given for 6 weeks and bassorin for 2 weeks to each of four piglets. A group of four piglets served as controls. Gallbladder bile was obtained by ultrasonically guided puncture of the gallbladder. The results showed no significant alteration of bile acids, lipid composition, and lithogenity of bile for either of the diets. These findings are interpreted in that way that the normally high basic fiber content of the food of piglets does not allow a further decrease of the lithogenic index by addition of wheat bran or bassorin. This conclusion would mean that the basic fiber diet should be known before therapeutic trials with fiber diets are planned or interpreted. PMID- 6281868 TI - Infection of sheep and goats with bovid herpesvirus 2. AB - Sheep and goats were shown to be susceptible to experimental infection with bovid herpesvirus 2 (BHV2), administered by either the intradermal or intravenous route. Lesions developing in sheep following intradermal inoculation of virus were similar to those in cattle inoculated intradermally, whereas the lesions in goats resolved without ulceration or scabbing. Disseminated circumscribed skin lesions developed in sheep and goats given BHV2 by the intravenous route. These lesions resolved in four to eight days without significant effect on the skin. BHV2 was isolated from skin lesions of sheep, goats and cattle that were infected intravenously, from sheep and cattle infected intradermally and from the leucocytes of the three species following intravenous inoculation of virus. Latent infection of sheep and goats was demonstrated following corticosteroid treatment 60 days after infection. PMID- 6281867 TI - An epidemic of swine influenza in Japan. AB - In a serological survey of pigs in Japan haemagglutination inhibition (HI) antibody to swine influenza virus (Hsw1N1) was first detected in 721 of 3313 (21.1 per cent) sera in 1977 and 1578 of 4946 (32.1 per cent) in 1978. The virus was prevalent in all districts of Japan but to a different extent. In contrast to the high incidence of antibody, there were few cases of infection with clinical signs and only 13 outbreaks of swine influenza were recognised in Japan from 1978 to 1979. Thirty-seven influenza viruses were isolated from nasal swabs of diseased pigs in 13 outbreaks. The subtype of most isolates was Hsw1N1 but two strains were Hsw1N2. PMID- 6281869 TI - Effects of dexamethasone infusion on the plasma cortisol response to cosyntropin (synthetic ACTH) injection in normal dogs. AB - Graded dosages of cosyntropin (synthetic corticotropin) were injected into groups of normal dogs on consecutive days. On the first day, cosyntropin was administered alone and, on the second, dogs were infused with dexamethasone three hours before cosyntropin injection. Adrenocortical function was assessed by sequential measurement of plasma cortisol (hydrocortisone) concentration. While no response differences were noted to the various amounts of cosyntropin injected with or without dexamethasone pretreatment, the magnitude of adrenocortical response was significantly greater in dogs infused with dexamethasone. It is concluded that dexamethasone pretreatment renders the canine adrenal cortex more responsive to a subsequent injection of cosyntropin. The combined dexamethasone infusion-cosyntropin injection test produces consistent adrenocortical responses in normal dogs, and has potential value in evaluation of adrenopathic dogs. PMID- 6281870 TI - Evidence for papillomaviruses in ocular lesions in cattle. AB - Negatively stained preparations of various ocular lesions, generally considered to be precursors to bovine ocular squamous cell carcinoma, were subjected to electron microscopic examination for viruses. Lesions examined included five conjunctival plaques, an acanthotic lesion from an eyelid, 15 conjunctival papillomas, two papillomas of the eyelid and two keratinised elongated proliferative lesions of the eyelid (cutaneous horns). Eight lesions contained particles that resembled papillomaviruses. These consisted of a conjunctival plaque, five conjunctival papillomas, a papilloma of the eyelid and one of the samples of cutaneous horn. The particles were present in small numbers and were found only after prolonged search. The finding of these particles suggests that papillomavirus may be involved in the aetiology of bovine ocular squamous cell carcinoma. PMID- 6281871 TI - [Clostridium perfringens in meat products in Chile: contamination level and enteric toxicity (author's transl)]. PMID- 6281872 TI - [Anti hepatitis A antibodies in healthy children and in patients with hepatitis (author's transl)]. PMID- 6281873 TI - [Chemotherapy of bronchial cancer: promise or reality?]. PMID- 6281875 TI - [Effect of physicochemical factors on ochratoxin A, and determination of its concentration in grain by spectrofluorometry. II. Selection of optimal conditions for thin-layer chromatography, method of ochratoxin A extraction and its concentration in grain]. PMID- 6281874 TI - Cytomegalovirus infections following renal transplantation. AB - This collective review of the literature concerns posttransplantation cytomegalovirus (CMV) infections. In the author's view, patients without previous CMV infection are most often infected by viruses transmitted with the transplanted kidney, whereas patients with prior infections can be infected either from this source or by reactivation of latent CMV. Many posttransplantation CMV infections are asymptomatic or yield only mild systemic effects. A few patients suffer life-threatening disease. However, CMV infections might adversely affect survival of both graft and patient by contributing to graft rejection, weakening the graft recipient's immunity, or by other, more direct, means. Currently, there is no effective, specific treatment of these infections. It is predicted that CMV infections will cease to be a problem only when the means are found to specifically block transplantation immunity, thereby eliminating the need for systemic immunosuppression and eliminating low-level host-vs.-graft responses. PMID- 6281876 TI - [Neuropharmacologic control of the secretion of hypophyseal hormones in man: dopamine, noradrenaline, serotonin. Clinical implications]. AB - The secretion of the pituitary hormones is controlled by hypothalamic hormones which are synthetised by neurosecreting cells whose activity is modulated by different neurotransmitters as dopamine, norepinephrine and serotonin. Centrally acting drugs (antiparkinson, antipsychotic, antidepressive and hypotensive agents) interfere with the activity of the neurotransmitters. Thus they may influence the secretion of hypothalamic and pituitary hormones. The action of different psychotropic drugs on the secretion of hormones of the anterior pituitary gland and of ADH is reviewed, taking into account differences existing between animal and man, with a special emphasis on: 1. the role of the neurotransmitters on the secretion of the hormones under physiological and pathological (endocrinopathies, neurological and psychiatric diseases) conditions; 2. the clinical importance of the secondary endocrinological effects that these drugs may elicit; 3. the efficacy of these drugs as therapeutic and diagnostic agents in endocrinopathies. PMID- 6281877 TI - [Viral diarrhea]. AB - Viruses are one of the most frequent causes of acute infectious gastroenteritis. Thus, rotaviruses are responsible for 40-60%--and in winter sometimes for over 90% - of diarrhoea in infants. These viruses may also cause disease in adults, but small viruses of a size of about 27 nm, such as the Norwalk agent, play a much more important role. Several other non-cultivable viruses such as adeno-, calici- and astroviruses have been implicated as a cause of infectious diarrhoea. Knowledge of rotaviruses is so far advanced that routine laboratory diagnosis can easily be performed and active immunization against rotavirus infection envisaged. PMID- 6281878 TI - [Glucagonoma syndrome in a multihormonal pancreatic tumor]. AB - A 60-year-old patient developed signs and symptoms of glucagonoma syndrome (dermatitis, weight loss, anemia and hypoaminoacidemia). However, diabetes mellitus was absent. Glucagonoma was suspected because of markedly elevated plasma glucagon levels and the tumor was subsequently removed by surgery. Acidethanol extraction of the tumor and immunohistochemistry provided evidence of the presence of all four islet hormones, particularly that of glucagon and pancreatic polypeptide and to a lesser extent of somatostatin and insulin. Immunohistochemistry of the tumor (but not plasma) also showed the presence of alpha-HCG. Plasma glucagon immunoreactivity consisted to a large extent (approx. 90%) of a high molecular form of glucagon, probably proglucagon. In spite of the presence of alpha-HCG - which is assumed to be a marker of malignancy - the patient has been free of recurrence for the 2 1/2 years since surgery. The increasing number of cases reported during the past few years demonstrates that the syndrome is more common than previously suspected. Glucagon secretion and its typical clinical picture may be a valuable marker of a multihormonal pancreatic tumor. In a case of suspected glucagonoma, diagnosis can be established simply by obtaining a plasma glucagon level measurement. PMID- 6281879 TI - Oncogenes. PMID- 6281880 TI - Residual calcium ions depress activation of calcium-dependent current. AB - Calcium ions enter and accumulate during depolarization of some cells, activating a potassium current, IK(Ca), that depends on the cytoplasmic concentration of calcium ions, [Ca]i. However, elevation of [Ca]i can depress IK(Ca) elicited by a subsequent membrane depolarization. The depression of IK(Ca) is ascribed here to a [Ca]i-mediated inactivation of the voltage-gated calcium conductance, which causes a net reduction in calcium ions available for the activation of IK(Ca). This suggests that other processes dependent on gated calcium entry may also be depressed by small background elevations in cytosolic free calcium ions. PMID- 6281881 TI - Autoradiographic evidence for a calcitonin receptor on testicular Leydig cells. AB - Previous studies have indicated that there is a relation between testicular function and adequate concentrations of zinc in testicular cells, and that calcitonin alters cellular zinc transfer in the testis. The present studies provide autoradiographic evidence that calcitonin binds in vivo to the cell membrane of testicular Leydig cells. The data thus confirm the presence of the testicular cell membrane calcitonin receptors that were previously demonstrated indirectly by Scatchard analysis of data collected from binding studies. PMID- 6281882 TI - Adenosine triphosphate synthesis coupled to K+ influx in mitochondria. AB - The influx of K+ into swollen mitochondria in the presence of valinomycin results in the synthesis of adenosine triphosphate in which approximately one H+ disappears per adenosine triphosphate synthesized. The synthesis is blocked by atractyloside but is insensitive to oligomycin and relatively insensitive to uncouplers. PMID- 6281883 TI - Cytomegalovirus antibody in cerebrospinal fluid of schizophrenic patients detected by enzyme immunoassay. AB - By means of enzyme immunoassay techniques to detect the presence of antibody to cytomegalovirus, the cerebrospinal fluid of 178 patients with schizophrenia, 17 patients with bipolar disorders, and 11 other psychiatric patients was compared with that of 79 neurological patients and 41 normal control subjects. The cerebrospinal fluid of 20 of the schizophrenic patients and 3 of the patients with bipolar disorders showed significant increases in immunoglobulin M antibody to cytomegalovirus; no difference was found in patients on or off psychotropic medications. PMID- 6281884 TI - Spectral character of sunlight modulates photosynthesis of previtamin D3 and its photoisomers in human skin. AB - The photosynthesis of previtamin D3 from 7-dehydrocholesterol in human skin was determined after exposure to narrow-band radiation or simulated solar radiation. The optimum wavelengths for the production of previtamin D3 were determined to be between 295 and 300 nanometers. When human skin was exposed to 295-nanometer radiation, up to 65 percent of the original 7-dehydrocholesterol content was converted to previtamin D3. In comparison, when adjacent skin was exposed to simulated solar radiation, the maximum formation of previtamin D3 was about 20 percent. Major differences in the formation of lumisterol3, and tachysterol3 from previtamin D3 were also observed. It is concluded that the spectral character of natural sunlight has a profound effect on the photochemistry of 7 dehydrocholesterol in human skin. PMID- 6281885 TI - Insulin stimulation of nucleoside triphosphatase activity in isolated nuclear envelopes. AB - The activity of nucleoside triphosphatase, an enzyme that regulates nuclear messenger RNA transport, was measured in highly purified nuclear envelopes isolated from rat liver. Addition of picomolar concentrations of insulin to freshly prepared nuclear envelopes directly increased the enzyme activity. The major effect of insulin on this enzyme was to increase the maximum velocity of its activity; no significant effects were seen on the affinity constant. These studies raise the possibility, therefore, that the nuclear envelope is a site where insulin regulates nuclear functions. PMID- 6281886 TI - Corticotropin-releasing factor stimulates accumulation of adenosine 3', 5' monophosphate in rat pituitary corticotrophs. AB - The presence of synthetic ovine corticotropin-releasing factor leads to a rapid and marked stimulation of adenosine 3', 5'-monophosphate accumulation in an enriched population of rat pituitary corticotrophs in primary culture. The increase, observed as early as 60 seconds after the addition of corticotropin releasing factor, suggests that changes in the intracellular concentration of the cyclic nucleotide coincide with or precede the secretion of adrenocorticotropic hormone in response to corticotropin-releasing factor. PMID- 6281887 TI - Intracellular pH of mitogen-stimulated lymphocytes. AB - Mitogenic stimulation of mouse lymphocytes results in two sequential intracellular alkalinizations. The first shift of intracellular pH from 7.18 to 7.35 coincides with early biochemical events following mitogenic stimulation. The second alkalinization begins 12 hours after stimulation and rises in parallel with the rate of thymidine incorporation. The results suggest that intracellular alkalinization following stimulation may play a key role in the enhancement of cellular activation and mitogenesis. PMID- 6281888 TI - Suppression of transcription termination by phage lambda. AB - The bacteriophage lambda N gene product positively controls development by preventing termination of transcription at terminator sites critical to the sequential expression of phage genes. Many host transcription factors, including RNA polymerase, are involved in N gene action. Recent findings have shown that ribosomal proteins are also involved. The current understanding of how the N protein affects transcription termination is reviewed, and a possible model and current problems are discussed. PMID- 6281889 TI - The Alu family of dispersed repetitive sequences. AB - A family of related sequences that includes approximately 500,000 members is the most prominent short dispersed repeat family in primate and rodent DNA's. The primate sequence is approximately 300 base pairs in length and is composed of two imperfectly repeated monomer units, whereas the rodent repeat consists of only a single monomer. Properties of this repeat sequence, its flanking sequences in chromosomal DNA, and RNA's transcribed from it suggest that it may be a mobile DNA element inserted at hundreds of thousands of different chromosomal locations. PMID- 6281890 TI - Isolation of human oncogene sequences (v-fes homolog) from a cosmid library. AB - To define the human homolog (or homologs) of transforming sequences (v-fes gene) common to Gardner (GA) and Snyder Theilen (ST) isolates of feline sarcoma virus (FeSV), a representative library of human lung carcinoma DNA in a cosmid vector system was constructed. Three cosmid clones were isolated containing GA/ST FeSV v fes homologous cellular sequences, within 32- to 42-kilobase cellular inserts representing 56 kilobases of contiguous human cellular DNA. Sequences both homologous to, and colinear with, GA or ST FeSV v-fes are distributed discontinuously over a region of up to 9.5 kilobases and contain a minimum of three regions of nonhomology representing probable introns. A thymidine kinase selection system was used to show that, upon transfection to RAT-2 cells, the human c-fes sequence lacked detectable transforming activity. PMID- 6281891 TI - Organization of endogenous opiate and nonopiate pain control systems. AB - Research during the past decade has revealed the existence of neural systems that modulate pain transmission. Much of this work has focused on the role of endogenous opiate systems, but recent research indicates the involvement of nonopiate mechanisms as well. In this article, we present data demonstrating that opiate and nonopiate analgesia systems can be selectively activated by different environmental manipulations and describe the neural circuitry involved. Both neural and hormonal pathways and both opiate and nonopiate substances play roles in the complex modulation of pain transmission. The existence and description of these modulatory mechanisms have important clinical implications for the treatment of pain. PMID- 6281892 TI - Interaction of convulsive ligands with benzodiazepine receptors. AB - The gamma-aminobutyric acid (GABA)-benzodiazepine receptor complex, which is composed of distinct proteins embedded in the neuronal plasma membrane, is important for several effects of benzodiazepines, including protection afforded against convulsions. During structural modification of ethyl beta-carboline-3 carboxylate an agent was discovered which has high affinity for brain benzodiazepine receptors but which is a potent convulsant. Also in contrast to benzodiazepines, this type of benzodiazepine receptor ligand favors benzodiazepine receptors in the non-GABA-stimulated conformation, which may explain the convulsive properties. PMID- 6281893 TI - Micromolar affinity benzodiazepine receptors: identification and characterization in central nervous system. AB - Receptors that selectively bind micromolar concentrations of benzodiazepines are present in rat brain membrane. These micromolar receptors exhibit saturable, stereospecific binding, and the potency of benzodiazepine binding to these receptors is correlated with the ability of the benzodiazepines to inhibit maximum electric shock-induced convulsions. Benzodiazepine receptors with nanomolar affinity differ from the micromolar receptors in their binding, kinetic, and pharmacologic characteristics. The micromolar receptors also bind phenytoin, a non-benzodiazepine anticonvulsant. These results provide evidence for a distinct class of clinically relevant benzodiazepine receptors that may regulate neuronal excitability and anticonvulsant activity. PMID- 6281894 TI - Use of "Super-plus" tampons discouraged. PMID- 6281895 TI - [New methods for computerized classification of psychopharmaceutical agents (author's transl)]. AB - We describe a statistical technique for achieving unbiased classification of 111 psychopharmaceutical agents from computerized clinical data. Three different methods were used: the reciprocal averaging method describes and pictures the clusterings of the numerous clinical datum available; the cluster analysis method corroborates the arrangement of the drugs and their effects into groups; the discriminating analysis method assigns the remaining drugs to their groups. The drugs were ultimately classified into six different groups. One of these groups, which included many of the antidepressant drugs, was reanalyzed with the reciprocal averaging method. This led to separating the monoamine oxidase inhibitors from the other antidepressants. Our technique will be immediately useful for classifying new psychopharmaceutical agents. If biologic data are included, this technique will be helpful for studying correlations between the structure and the effects of drugs. PMID- 6281896 TI - [Congenital paramyotonia, adynamia episodica hereditaria, or familial periodic paralysis with paramyotonia and hyperkalemia (author's transl)]. AB - Case-reports of eleven patients diagnosed either as Eulenburg paramyotonia or Gamstorp adynamia episodica hereditaria are reviewed. The connections between both conditions are discussed. The clinical pictures are similar, with predominance of either paramyotonia or adynamia. In both conditions, hyperkaliemia is present and potassium loading may be followed by clinical exacerbation. Electromyographic and histological findings are comparable. Paramyotonia and adynamia probably result from the same physiopathological mechanism : hyperkaliemia modifies the cell membrane permeability to sodium and potassium, leading to depolarization with initial hyperexcitability and subsequent inexcitability. We believe that Eulenburg paramyotonia and Gamstorp adynamia episodica hereditaria are two manifestations of the same condition which could be termed familial periodic paralysis with paramyotonia and hyperkaliemia. PMID- 6281897 TI - [Advances in diagnosis and treatment of oligodendrogliomas of the brain. Eighty four surgical patients (author's transl)]. AB - The authors report their experience of 84 patients with oligodendroglioma seen over a 34 year period. The low incidence of these brain tumors is underlined. Oligodendroglioma is the most epileptogenic of brain tumors. The clinical picture may resemble "essential" epilepsy. Normal EEG should be considered suspect in confirmed epileptic children and adolescents since oligodendroglioma is the most common of all tumors which induce long-standing epilepsy. Emphasis is put on the importance of computerized tomography and radionuclide scanning for early detection of these tumors. Both procedures are very helpful for deciding on the best operative approach and for determining whether the lesion is malignant or not. The authors advise tumorectomy, which should be as complete as possible, followed by systematic radiotherapy, even if histopathological criteria indicate a benign tumor. With this therapeutic regimen mean survival rates are significantly higher than those published previously. PMID- 6281898 TI - [Parkinson disease and depression (author's transl)]. AB - Depression and Parkinson disease, two very different conditions at first sight, have much more intricate connections than is usually believed. Depression may be the patient's reaction to Parkinson disease, a condition that is anticipated with anxiety, with good reason as it is often very disabling and has not been significantly prolonged by dopamine therapy. Depression may precede the first signs of Parkinson disease. Pseudoparkinsonian melancholia may be difficult to distinguish from the akinetic form of Parkinson disease. Most of the symptoms of the latter have been encountered in the former. The following features do not occur in depression: astasia with trepidation, festination, monotonous tachyphemia and palilalia, sebaceous hypersecretion, and of course unilateral or frankly asymmetric signs. Parkinson syndrome secondary to depression can be classified with those parkinsonian syndromes that are different from parkinson disease and secondary to a clearcut etiology. In some instances, diagnosis is established by the response to therapy. In the present state of our knowledge, the treatment of depression relies on chemotherapy and sismotherapy and not on dopamine therapy. The management of Parkinson disease rests on dopamine which may be associated with tricylic antidepressants. PMID- 6281899 TI - [Practising psychiatry in a non-psychiatric hospital department: description of our experience (author's transl)]. AB - The author describes a practical set-up for practising psychiatry in a non psychiatric department of a general hospital. Initially, patients were seen on an unorganized basis, each time psychiatric advice was asked for. Subsequently, a meeting was held each week, with all the staff members (physicians, nurses, social workers...) and the same consulting psychiatrist. This meeting serves several purposes: clinical evaluation of patients, training of staff members, exchange of information between hospital departments, and choice of treatment. The different situations leading to admission and the diversity of clinical pictures are described. Appropriate answers are more likely to be provided if each staff member is assigned clear-cut responsibilities. PMID- 6281900 TI - [A clinical trial of tiapride in neurology (author's transl)]. AB - Results in twenty-three patients given tiapride are reported on. As an analgesic, tiapride's efficacy was questionable. In patients with delirium, sedation was not completely satisfactory; however, this may be due to the small number of patients and to the low doses given. The excellent results recorded in patients given tiapride for extrapyramidal hyperkinesia due to various etiologies are consistent with previous reports in the medical literature. PMID- 6281901 TI - [Spinal subdural hematoma. An unusual complication of lumbar puncture (author's transl)]. AB - Spinal subdural hematoma following lumbar puncture occurred in a patient with embolic hemiplegia. The authors discuss the possible responsibility of heparin administered immediately after the tap. Physiological mechanisms of spinal subdural hematoma are reviewed. The authors emphasize the poor prognosis associated with this rare complication. They suggest that anticoagulant therapy be discontinued during and after a spinal tap. The first signs of medullary and radicular compression should be promptly recognized and an emergency neurosurgical procedure undertaken in order to forestall definitive paraplegia. PMID- 6281902 TI - [Approach of the family in general hospital psychiatry (author's transl)]. AB - The new theoretical models propounded for understanding family relationship dynamics are very useful in psychiatric hospital care. In our paper two clinical observations exemplify how a systematic understanding of psychiatric disorders, which replaces the symptom within the family system, and an individual psychodynamic understanding can be combined, and how interviews with the whole family group can help in setting up a therapeutic program which the patient perceives clearly. We believe that, with this method, the patient's and his family's abilities for change and care are best availed of. PMID- 6281903 TI - [Alcoholism in young people (author's transl)]. AB - Epidemiology should be used for studying teenagers' attitudes and behaviours concerning alcoholic beverages in order to develop health education programs designed to induce temperance in adolescence and, subsequently, in adulthood. We report on five studies carried out in France, including 5,481 subjects, both male and female, aged 13 to 26, belonging to various communities, and living in different conditions. Answers to questionnaires showed that both the first ingestion of alcohol (usually with the family) and the first inebriation (usually with friends) occur very early (at ages 10 to 12 and 15 to 16 respectively). Alcohol intake increases with age, parents' permissiveness, money allowance, and pub attendance. Health education should be undertaken very early, in kindergarten, and subsequently included in school programs each year. Cooperation among school teachers and with the school's medical staff and the childrens' parents is essential. PMID- 6281904 TI - [Anaplastic carcinoma of the thyroid: early clinical and histopathological stages. Therapeutic implications (author's transl)]. AB - Anaplastic carcinoma of the thyroid is generally considered a primary condition. Our study, based on ten observations, shows that anaplastic carcinoma of the thyroid with a rapidly fatal course may arise in long-standing goiters. Borderline histologic changes may be encountered; in some differentiated nodules, undifferentiated areas were found. Factors inducing transformation is equally obscure. Anaplastic changes may result from the growth of previously quiescent clones. The time-interval between benignant and malignant lesions varies widely and is difficult to establish unequivocally. In some instances, anaplastic carcinoma occurs after surgical removal of differentiated carcinoma. PMID- 6281905 TI - [Surgical treatment of acute complicated colitis. Report on twenty-eight cases (author's transl)]. AB - Experience with 28 patients with acute diffuse complicated colitis operated on in emergency or semi emergency by the same surgical team is reviewed. The forms with colonic dilatation are the most numerous but do not resume the serious complications which may occur in the course of non specific inflammatory diseases of the colon. The operative mortality in this series was 10,7% (3/28). Peritoneal sepsis was the most significant factor contributing to mortality. It appears that the keystone to successful management is to prevent colonic perforation. Protracted medical management may be at last partly responsible for this complication. Failure of intensive medical therapy to induce rapid improvement constitutes an indication for definitive surgical treatment. PMID- 6281906 TI - [Conservative treatment of breast carcinoma with or without prophylactic chemotherapy. Preliminary results in 45 cases (author's transl)]. AB - Between February 1969 and December 1979, 45 breast carcinomas T1 NO or T2 NO (greater than 3cm) were treated by tumorectomy and radiotherapy. Axillary dissection was performed in 22 cases. Twenty-one pre-menopausal patients were castrated either by oophrectomy (13 cas) or by pelvic irradiation (8 cases). From 1977 on, (10 cases) prophylactic chemotherapy using cyclophosphamide, 5 FU, methotrexate, vincristine, was administered for 4 to 6 months according to the number of poor prognostic factors. The five-year actuarial remission rate was 93% (84-100), the metastatic remission rate was 88% (72-100). Indications of prophylactic chemotherapy in forms with good prognostic factors are discussed. PMID- 6281907 TI - [Non typhoidal Salmonella infections: a clinical study of 491 cases (author's transl)]. AB - 491 cases of non typhoidal salmonella infections were studied in the Rouen Hospital University Centre. Affected patients are mainly children who usually present mild febrile gastroenteritis. Severe forms with septicemia, enteric fever, or focal manifestations, usually occur in adults. In nearly half the cases an underlying disease or another predisposing condition was found. In these cases, the mortality rate was increased. Antibiotic therapy prolonged post convalescent excretion of salmonellae. Adequate diet and hygiene are sufficient in mild forms. Antibiotics should be used only in acute salmonella infections. PMID- 6281908 TI - [Metacarpophalangeal calcifications in diffuse chondrocalcinosis. Radiologic study of ten cases (author's transl)]. AB - The authors report ten cases of metacarpophalangeal calcifications in chondrocalcinosis. These calcifications are associated with a calcification of the triangular ligament of the wrist in the all cases. Cartilaginous calcifications of the wrist are found in 9 cases. Metacarpophalangeal calcifications concern mostly the third (9 cases) and the second (6 cases) metacarpophalangeal joint. They are present frequently on radial side (29 joints) rarely on cubital side (5 joints) with a calcification on radial and cubital side in 4 joints and a calcification on cubital side only in one joint. Calcic deposit concern almost the synovial and capsule, it is rarely cartilaginous one. The authors discuss the possibility of calcic deposit of the glenoidal fibrocartilage of the metacarpophalangeal joint. PMID- 6281909 TI - [Pseudohypoaldosteronism: familial forms. Eight case-reports, with a review of the literature (author's transl)]. AB - Pseudohypoaldosteronism (PHA) is an uncommon cause of the renal salt loosing syndrome in infancy. The authors describe eight cases in two different families. Hyperaldosteronism persists long after clinical recovery has occurred. Plasma hormone assay allows retrospective recognition of cases overlooked during infancy. This underlines the variability of disease expression among different members of the same family. The high family occurrence rate (over 50%), which is often underestimated, is demonstrated by the study of both families and by a review of published cases. Clinical and biochemical features of familial PHA are discussed. Inheritance is usually on an autosomal dominant basis. However, the small number of reported cases cannot allow any attempt to individualize subgroups of the disorder upon genetic grounds. PMID- 6281910 TI - [Cancer of the kidney in adult. Report of an homogeneous series of eighty cases (author's transl)]. AB - The authors report their experience of cancer of the kidney in adults involving a homogeneous series of eighty operated cases. They draw from this experience several conclusions concerning the clinical, diagnostic, therapeutical and prognostic aspects and underline three points; the interest of arteriography combined with embolization, the significance of an enlarged nephrectomy without lymphatic evidement, carried out with the least damage possible, and the abandoning of pre- and postoperative radiotherapy. Results seem encouraging since the survival rate reaches respectively 87,5 p. cent after five years and 66,9 p. cent after ten years for stage I, 55,5 p. cent after five years and 40 p. cent after ten years for stage II. PMID- 6281911 TI - [Spontaneous peritonitis in a cirrhotic patient (author's transl)]. AB - A case of spontaneous peritonitis in a cirrhotic patient is reported. Because of marked peritoneal and hemodynamic signs, exploratory laparotomy was decided on. Such cases of spontaneous peritonitis, which mimick surgical affections, are very uncommon. Onset is usually insidious with increasing hepatic fluid as the only sign. Bacteriologic examination of the peritoneal fluid is often negative. Diagnosis should be considered if cytology shows more than 75 polynuclear leukocytes per mm. When bacteriology is negative, association of an aminoside with penicillin G and metronidazole seems to be the most satisfactory combination. PMID- 6281912 TI - [Subdural empyema: a rare complication of ENT infections in childhood. Two case reports (author's transl)]. AB - Two cases of subdural empyema are reported. One occurred after sinusitis, the other after otitis media. In both cases, the diagnosis was suggested by a combination of symptoms indicating infection and neurological disease, which occurred soon after treatment of an ENT infection. Diagnosis was established, either by CAT scan, or by arteriography. Treatment consisted of surgical drainage and optimal antibiotherapy. The clinical course was favorable: complete recovery took place in both cases and clinical status remains unremarkable after two years. The current literature is reviewed and the main clinical features and diagnostic tools (especially CAT scan) are recalled. PMID- 6281913 TI - [Short and long term results in the treatment of obesity. A study in 173 patients (author's transl)]. AB - Short and long term weight loss was studied in 173 obese patients treated by a low-calorie diet. Significant weight loss was observed in 78% of the patients after 6 months and in 58% after 4 years. These results are similar to those published in other studies but the comparison is difficult because the methods used are not strictly comparable. A detailed study of 116 patients showed that weight loss is not significantly influenced by such variables as age, sex, previous calorie intake, complications, and severity or duration of the obesity. PMID- 6281914 TI - Radiopharmaceuticals for hepatobiliary imaging. AB - Tests for liver function have by and large centered around clinical laboratory diagnostic procedures for a number of years. Besides these, radiographic imaging procedures, including oral cholecystography and intravenous cholangiography, serve a very useful purpose, but several of them are invasive and involve a certain degree of risk from the administered contrast media as well as discomfort to the patient. The cholescintigraphic procedures, though noninvasive, have not played a significant role in the evaluation of hepatobiliary disorders prior to the introduction of the currently available. 99mTc-labeled IDAs. These new hepatobiliary agents offer many advantages over the previously utilized radiopharmaceuticals (131I-rose bengal in particular) in terms of the high degree of specificity for localization in the gallbladder with rapid extraction rates by the polygonal cells of the liver and very low excretion via the GU tract. A detailed understanding of the structure distribution relationship of the various groups in the complex enable the design of agents with an improvement in hepatobiliary specificity and other desirable characteristics. In many clinical situations, even in patients with high bilirubin levels, the 99m-Fc-labeled IDAs offer far superior clinical information over the alternative diagnostic imaging modalities. Further, the absorbed radiation dose imparted to the critical organs is far lower than with the older agents. Thus, the introduction of the cholescintigraphic procedures with the 99mTc-labeled IDAs have ushered in a new phase in the diagnostic workup of patients with impaired hepatocellular function and other biliary disorders. PMID- 6281915 TI - Current status of radiocolloid hepatic scintiphotography for space-occupying disease. PMID- 6281916 TI - Liver uptake of 99mTc-pyrophosphate. PMID- 6281917 TI - Meckel's diverticulum. PMID- 6281918 TI - Social factors in the etiology of chronic disease: an overview. AB - Our intention in this introductory article is to emphasize what we consider to be certain critical points in the current state of research into the social epidemiology of chronic disease. As will be outlined, these critical points need to be considered in future research. To begin with we provide a mild critique of research in this area which has had its tradition specified by social epidemiology, a term which has recently come into favor to describe research concerned with social factors in the etiology of chronic disease. Next we briefly summarize critical available evidence on an etiological relationship between social factors and cardiovascular disease, cancer and multiple disease outcomes. Following this a major emphasis will be placed on issues which directly relate to problems of methodology in social factors assessment. Finally, an emphasis will be given to the critical question of mechanisms which need to be clarified in this type of multivariate research. PMID- 6281920 TI - pneumocystis carinii Pneumonia in a male homosexual. PMID- 6281919 TI - The relation of social to biological processes in disease. AB - A common language is developing which describes how social interaction can lead to disease. There is a consensus based on forty years of psychophysiological research that emotion is the crucial driving force in a chain of events leading from psychosocial interaction to neuroendocrine changes. These changes, in turn, can induce physiological abnormalities eventually leading to pathologic changes. The emotions involved have been presented in a model in which a sympathetic adrenal-medullary effort-relaxation axis is contrasted with a pituitary adrenal cortical elation-dejection axis. A further pituitary-gonadotropic axis for social success as opposed to low social assets is described. Although overlapping, each system can be shown to be related to a separate neuroendocrine mechanism whose activity pattern may be either exaggerated or suppressed. Social scientists like Kemper and gender psychologists like Bakan speak of separate power (agentic) and status (communion) axes. They appear to correspond broadly to the neuroendocrine mechanisms mentioned earlier. The changes induced by emotions are sufficiently powerful to regularly override the neuroendocrine feedback controls that should restore homeostasis. These overrides will either gradually or sometimes abruptly- as in cardiac arrhythmia--lead to pathophysiological changes and so to disease states, which are eventually fatal. Although social supports and social assets are successful in keeping neuroendocrine disturbances to a minimum in a healthy society, if the milieu is disordered, one's vulnerability is increased. PMID- 6281921 TI - Surveillance of Japanese encephalitis cases in Thailand. AB - This surveillance study reveal the incidence of Japanese encephalitis cases throughout the year with an increase during the rainy-seasons. JEV attacks all ages, but with a high incidence found in age group between 8 to 14 years determined from the studies in 1974, 1975 and 1976 and records of distribution in 40 provinces of Thailand. The severity of infection is high to warrant virus surveillance by the Ministry of public Health. PMID- 6281922 TI - Passively acquired antibody to flavivirus in Thai infants. PMID- 6281924 TI - Serum angiotensin-converting enzyme estimation and the Kveim-Siltzbach test in sarcoidosis. AB - Serum angiotensin-converting enzyme (SACE) and Kveim-Siltzbach test results were obtained in 25 patients with clinically active sarcoidosis. Twenty of these patients had elevated SACE levels and 17 had positive Kveim-Siltzbach tests. One patient had normal SACE activity and a negative Kveim-Siltzbach test. Four of the 6 patients with a normal SACE level had a positive Kveim-Siltzbach test and 7 of the 8 patients with a negative Kveim-Siltzbach test had elevated SACE levels. These results indicate that the two tests are complementary and increase the diagnostic specificity in sarcoidosis. We conclude that the concurrent use of SACE measurement and Kveim-Siltzbach tests is of value in the diagnosis of sarcoidosis. PMID- 6281923 TI - Cellulose acetate membrane precipitin (CAP)test in amoebiasis using antigens from four different strains of Entamoeba histolytica. AB - A simple cellulose acetate membrane precipitin (CAP) test was evaluated against immunoelectrophoresis (IEP) test using saline extract from 4 different strains (HK-9, HT-10, HT-12 and HT-31) of axenically grown Entamoeba histolytica as the antigens. All 81 sera from patients with amoebic liver abscess were positive in the IEP against the antigens from all 4 strains. With the CAP test the number positive against antigens from HK-9, HT-10, HT-12 and HT-31 were 79,77,79 and 71 respectively Sera from 100 blood donors were negative by both IEP and CAP tests against antigens from all 4 strains. Comparison between the number of precipitating bands demonstrated by either IEP or CAP test showed that strain HT 12 was the best source of antigen in exhibiting significantly more number of precipitating bands, strain HT-31 was the poorest. The strain HT-10 was comparatively superior to strain HT-31 in the CAP test whereas in the IEP test strain HK-9 and HT-31 were both inferior to strain HT-10. PMID- 6281925 TI - Intra-osseous synovial sarcoma. AB - Intra-osseous synovial sarcoma is very rare and its mode of origin is conjectural. Our patient's tumour was situated within the lower femur. The sited of origin of the few intra-osseous synovial sarcomas so far documented may support the concept of intra-osseous synovial rests. PMID- 6281926 TI - Vaccines for the decade of the 1980s and beyond. AB - An increase in the average human lifespan from ancient times to the present is due mainly to control of infectious diseases, and vaccines are a keystone to their prevention. Several important new vaccines should emerge during the present decade, the most important being those against human hepatitis A and B. Disease eradication and elimination by vaccines are worthwhile targets for at least some of the more important diseases and will probably be accomplished during this century. Further increase in lifespan should be achieved by vaccination in this century but major future advances will need to come from slowing the process of ageing. PMID- 6281927 TI - Human polyomavirus in herpes keratitis lesions following renal transplantation. PMID- 6281928 TI - Pathological correlation of CT ring in recurrent, previously treated gliomas. AB - The meaning of the enhancing ring commonly seen on computerized tomographic (CT) scans in patients with recurrent, treated glioblastoma has long been of concern. Does the ring represent radiation necrosis, pseudocapsule formation, compressed adjacent brain, or, in fact, a peripheral zone of viable tumor? We employed a multiple intratumoral biopsy technique at the time of reoperation to investigate this ring. Pathological evidence is presented documenting the content of the ring to the viable tumor. It is suggested that perhaps treatment failure may be determined on sequential CT examinations by comparing the volume and degree of contrast enhancement. PMID- 6281929 TI - [Combined craniomaxillo-facial resection for advanced malignant tumors of maxillofacial regions]. PMID- 6281930 TI - Intrahepatic abscess as a complication of long-term percutaneous internal biliary drainage. AB - One hundred five patients with obstructive jaundice have undergone percutaneous transhepatic internal biliary drainage at the Johns Hopkins Hospital. Many of these patients subsequently underwent corrective or palliative surgery, whereas other died of malignant disease after relatively short periods of catheter decompression, Seven of these patients with percutaneous internal biliary drainage, however, have been followed for over 8 months. Three of these seven patients developed intrahepatic abscesses at a mean of 16 months after catheter placement. Two of the three patients died of sepsis. In two of the patients the abscesses communicated with the biliary tree, in the third it did not. Intrahepatic abscess formation may be a common complication of long-term percutaneous transhepatic internal biliary drainage, and it should be suspected in any patient with fever or signs of sepsis who has been followed with catheter drainage for over 6 months. PMID- 6281931 TI - Adductor muscle group excision. AB - A surgical technique for removal of the adductor muscle group invaded by primary soft tissue sarcoma is described. A review of the clinical histories of 14 patients showed that the major complication of this procedure was prolonged suction catheter drainage of lymph, followed by sepsis along the drain tracts. No rationale for removal of the inguinal lymphatics in the absence of direct involvement by tumor or palpable lymphatic metastasis was found in the literature. We conclude that in primary soft tissue sarcoma of the adductor muscle group, clinically uninvolved superficial inguinal lymphatics should be carefully preserved. The limited benefits from removal of these lymph nodes does not merit the marked increase in operative morbidity. A surgical technique for adductor muscle group excision is presented that allows the best principles of sarcoma surgery to be practiced and that carries on acceptable operative morbidity rate. PMID- 6281932 TI - A case of insulinoma and approach to diagnosis. PMID- 6281933 TI - Angiotensin converting enzyme activity and evolution of pulmonary vascular disease in rats with monocrotaline pulmonary hypertension. AB - We have investigated the role of angiotensin converting enzyme (ACE) in the development of pulmonary hypertension, right ventricular hypertrophy, and pulmonary vascular disease in rats given a single subcutaneous injection of the pyrrolizidine alkaloid monocrotaline. Thirty-six young female Wistar rats were divided into a test group of 27 animals and a control group of nine animals. Each test rat was given a single subcutaneous injection of monocrotaline (60 mg/kg body weight). On the first, third, fifth, seventh, tenth, twelfth, fourteenth, seventeenth, and twenty-second days after the injection of monocrotaline the mean right ventricular systolic blood pressure was measured in one control and three test rats. The animals were then killed and we measured the specific activity of ACE in serum and lung homogenate. We also evaluated muscularisation of pulmonary arterioles, medial hypertrophy of muscular pulmonary arteries, and right ventricular hypertrophy. The sequence of changes was as follows: muscularisation of pulmonary arterioles and medial hypertrophy of muscular pulmonary arteries were apparent seven days after administration of monocrotaline; pulmonary hypertension and reduced lung ACE activity occurred after 10 days; right ventricular hypertrophy was detected after 12 days. Serum ACE activity was unchanged. It is concluded that the reduction in lung ACE activity is a result rather than a cause of the pulmonary hypertension. This reduction in lung ACE activity may be a protective mechanism designed to limit the elevation of the pulmonary arterial pressure. PMID- 6281934 TI - One very rare and one new tracheal tumour found by electron microscopy: glomus tumour and acinic cell tumour resembling carcinoid tumours by light microscopy. AB - Tracheal tumours were removed surgically from two patients and diagnosed as carcinoid tumours by routine light microscopy. At a later date, electron microscopy was performed on stored tumour tissue and no neurosecretory granules were found in either case. One showed features of a glomus tumour and the other of an acinic cell tumour. Only two glomus tumours appear to have been reported previously in the trachea, and no acinic cell tumours. Electron microscopy is thus sometimes of great assistance in diagnosing accurately unusual tumours of the lower respiratory tract. PMID- 6281936 TI - [Vaccination of cattle and sheep with M. K. 25]. PMID- 6281935 TI - [Perhexiline maleate induces sphingomyelinase deficiency in man]. PMID- 6281938 TI - Neurotoxins as probes in the study of neuronal development. AB - We have investigated the expression of surface membrane binding sites for tetanus toxin and alpha-scorpion toxin (AaHII) on cells of the in vivo developing mouse nervous system. There is a close temporal correlation in the pattern of emergence and accumulation of tetanus toxin binding cells (TBC) and that of post-mitotic neurons. In different nervous system areas, the fluctuations in relative TBC abundance reflect regional changes in the dynamics of neuronal subpopulations. The results indicate that the acquisition of membrane tetanus toxin binding sites may represent one of the earliest detectable characteristics of nascent neurons. The Na+ channel-associated scorpion toxin become detectable in fetal mouse brain two days after the appearance of TBC. Their density increases with fetal age without change in receptor properties. At all stages, scorpion toxin binds to a single class of noninteracting sites with a KD = 0.1 - 0.5 nM. The affinity of binding is voltage-dependent. Studies on brain cells and various cell lines grown in vitro suggest a selective association of the high affinity scorpion toxin receptors with neuronal phenotype. In culture, as in vivo, there is a time dependent increase in receptor density. These results indicate that both tetanus toxin and scorpion toxin can be used as qualitative markers of neuronal differentiation; moreover, estimates of the density of scorpion toxin binding sites provide a quantitative index of neuronal maturation. PMID- 6281937 TI - Electrophysiological analysis of the presynaptic action of beta-bungarotoxin in the central nervous system. AB - In view of previously reported actions of beta-bungarotoxin (beta-BuTX) on central brain synaptosomes, the effects on this toxin on the electrical activity of two brain slice preparations have been examined in vitro. beta-BuTX initially suppresses the synaptic component of the field responses to electrical stimulation in olfactory cortex and hippocampal slices. Intracellular recordings demonstrate that this synaptic depression occurs without detectable reduction in the sensitivity of the postsynaptic neuron to putative neurotransmitters. Following longer exposure to the toxin, reduced neuronal excitability is observed both pre- and post-synaptically. Elimination of the phospholipase A2 activity of beta-BuTX, by chemical modification or removing the Ca2+ necessary for enzymic activity, greatly reduces but does not totally eradicate the toxin's ability to block neurotransmission in the olfactory cortex. In the absence of enzymic activity beta-BuTX has no obvious effect on axonal conduction. Pure phospholipases A2, such as that from Naja melanoleuca mimic the transmission blocking action of beta-BuTX, but with lower potency and without the effects on fibre excitability. Collectively, these results are taken as evidence that beta BuTX initially suppresses transmitter release, a notion supported by the observed loss of spontaneous synaptic activity in hippocampal cells. Prolonged exposure to the toxin induces apparently less specific effects on neuronal excitability which are dependent on phospholipase A2 activity and are discussed with reference to the selective action of beta-BuTX on hippocampal fibre systems which possess release sites. PMID- 6281939 TI - Genetics and pathogenic role of Escherichia coli haemolysin. AB - While clear evidence exists for the direct involvement of cytolysins in the pathogenesis of Gram-positive bacteria, the significance of Gram-negative haemolysins remains unclear. This paper presents briefly data indicating a role for haemolysin production in infections caused by Escherichia coli and also experiments which have allowed an analysis of the molecular basis of the haemolysis among pathogenic and non-pathogenic strains of this species. PMID- 6281940 TI - Molecular Properties of neurotoxin receptors sites associated with sodium channels from mammalian brain. AB - Neurotoxins that act at specific receptor sites on voltage-sensitive sodium channels have been used as molecular probes to identify and purify protein components of sodium channels from mammalian brain. Photoreactive derivatives of scorpion toxin have been prepared and used to covalently label sodium channels in intact synaptosomes. Two polypeptides, alpha with Mr approximately 270,000 and beta with Mr approximately 38,000, are specifically labeled indicating that they are components of the scorpion toxin receptor site on the sodium channel. The sodium channel can be solubilized with retention of specific binding of [3H] saxitoxin using nonionic detergents such as Triton X-100. The solubilized saxitoxin receptor has molecular weight of 316,000 +/- 63,000 and binds 0.9 g of Triton X-100 and phospholipid per g of protein. The solubilized receptor can be purified 750-fold by ion exchange chromatography, wheat germ lectin/Sepharose chromatography and sucrose gradient sedimentation to a final specific activity of 1488 pmol/mg. Analysis of the polypeptide chain composition of the most highly purified fractions indicates that alpha and beta comprise 65% of the protein of these fractions and are only the polypeptides whose presence correlates with saxitoxin binding activity. These studies lead to a working hypothesis of sodium channel structure in which the intact channel is comprised of a complex with Mr of approximately 316,000 containing one mole of alpha (Mr approximately 270,000) and one to three moles of beta (Mr approximately 38,000). PMID- 6281942 TI - Effect of the venom of the snake Cerastes cerastes (African desert horned viper) on the response of the rat vas deferens to field stimulation. AB - Cerastes cerastes venom in low concentrations (0.8-2.5 micrograms/ml) inhibited the response of the rat vas deferens to field stimulation, while higher concentrations (4-8 micrograms/ml) causes a contracture that was reversed by washing and transiently blocked by phentolamine. The effect of the venom and guanethidine (10(-7) M) was reversed by tyramine (4.6 x 10(-8) M) but unaffected by hexamethonium (1.4 x 10(-7) M). When the response to field stimulation was inhibited by the venom the response to norepinephrine was either potentiated or unchanged. PMID- 6281941 TI - Two types of scorpion receptor sites, one related to the activation, the other to the inactivation of the action potential sodium channel. AB - The action of the neurotoxin in Buthinae scorpion venoms (Androctonus, Buthus or Leiurus genera) has been extensively studied. These proteins induce a prolongation of the action potential of nerves and muscles by slowing down inactivation of the sodium channel. Their affinity for their receptor site depends on membrane potential. In the present report we describe a toxin from a Centrurinae scorpion, Centruroides suffusus, which binds rat brain synaptosomes at a receptor site distinct from the Buthinae scorpion site independently of voltage. We name Androctonus-like toxins, alpha-scorpion toxins (alpha-ScTX), and Centruroides-like toxins, beta-scorpion toxins (beta-ScTX). We further report that beta-ScTX induces repetitive firing in frog myelinated nerve fibres by producing an abnormal sodium permeability. The beta-toxin binds specifically to rat brain synaptosomes (Kd = 3 nM) and induces an inhibition of the uptake and a stimulation of the release of GABA at concentrations which are in good agreement with the Kd value. These effects are blocked by tetrodotoxin. The binding site of beta -ScTX is distinct from those of other neurotoxins acting on the sodium channel like tetrodotoxin, alpha-ScTX and veratridine. The alpha-ScTX/beta-ScTX binding site capacities decreases as development of rat brain synaptosomes progresses ; at day 7 after birth, it is 1.1. and at day 39, 0.3. PMID- 6281943 TI - Sodium channels blocked by aphantoxin obtained from the blue-green alga, Aphanizomenon flos-aquae. PMID- 6281944 TI - In vitro effect of mercury on tissue phosphatases of Notopterus notopterus and the role of EDTA and AA in their restoration. AB - In vitro effects of four sublethal concentrations (0.10, 0.40, 0.80 and 1.20 micrograms) of mercury (hg) on acid, alkaline and glucose-6-phosphatases of liver, kidney and gills of Notopterus notopterus were studied. The maximum (80.76%) significant (P less than 0.001) inhibition in acid phosphatase of gills and minimum (36.66%) significant (P less than 0.05%) inhibition in alkaline phosphatase of kidney was observed at 1.20 microgram and 0.10 microgram Hg concentrations, respectively. EDTA and ascorbic acid (AA) restored the phosphatase activity to the greatest extent when administered with the Hg. PMID- 6281946 TI - Adenyl cyclase activity following fluoride ingestion. AB - Adenyl cyclase activity and fluoride content were investigated in various tissues of adult rabbits given NaF (10 mg/kg body wt./day) orally for 6 months. The activity of adenyl cyclase increased significantly in bone, liver, and kidney, whereas it was unchanged in skeletal muscle following fluoride ingestion. The levels of fluoride were elevated in serum and all the tissues investigated following fluoride ingestion; bone revealed the highest percentage increase and skeletal muscle the lowest. PMID- 6281945 TI - Enhancement of rat and human phagocyte superoxide anion radical production by cadmium in vitro. AB - The mechanism by which cadmium produced oxidizing effects in vivo is unknown. We show that cadmium enhances the production of superoxide anion radical (O-(2) .), a reactive oxygen species, in digitonin-stimulated phagocytes from man and rat. Cadmium concentrations ranging from 3.6 X 10(-2)M to 3.6 X 10(-4)M inhibited O (2) . production in rat alveolar macrophages or human granulocytes. However, when activated in the presence of 3.6 X 10(-5)M cadmium, the production of O-(2) . was increased by a factor of 2.11 +/- 0.25 above control levels in human granulocytes and 3.6 +/- 0.62 above control levels in rat alveolar macrophages. This effect by levels of cadmium within the range of those occurring during in vivo toxicity might provide an explanation for the oxidizing effects of this metal ion. PMID- 6281947 TI - Effects of a few xenobiots on three phosphatases of Saccobranchus fossilis and the role of ascorbic acid in their toxicity. AB - The effects of 0.0021, 0.00126 and 0.00063 mg thiotox per liter and 5.18, 3.11 and 1.56 malathion per liter (1/3, 1/5, and 1/10 fractions of 96-h LC50) on acid, alkaline and glucose-6-phosphatases in liver, brain and gills of Saccobranchus fossilis were studied after 30 days exposure with or without ascorbic acid. These enzyme activities were altered by these pesticides, but lesser alterations were observed when ascorbic acid was given. PMID- 6281948 TI - Aflatoxins, B1, B2, G1, G2 in primary liver cell carcinoma. AB - Aflatoxin B1, B2, G1, G2 levels were determined in sera of 20 patients with primary liver cell carcinoma (CC). Values of B1 above 0.15 microgram/ml were obtained in sera of three patients. Hepatitis B surface antigen (HBsAg) was detected in only 35% of patients. 50-90% of serum levels were cleared in 24 h in three patients studied. Although it is inconclusive as to the carcinogenic potential of aflatoxins in adult man, foodstuffs should be protected from excessive aflatoxin contamination. PMID- 6281949 TI - Clinical aspects of bile acid metabolism in liver diseases. AB - For a long time, it has been assumed that the liver plays an important role in the formation and excretion of bile acids and that the metabolism of bile acids, therefore, can be disturbed to some extent in liver diseases. Pathophysiological aspects of the disturbed metabolism of bile acids remained obscure at the time when procedures of qualitative and quantitative analyses, identification and estimation of bile acids began to be utilized in practice 20 years ago. Since then, the metabolism of bile acids has been clarified in individuals under normal as well as pathological circumstances. Because of a functionally and morphological close relation between hepatic epithelial cells and the metabolism of bile acids, the pathophysiological aspects of the disturbed metabolism of bile acids in parenchymatous inflammatory diseases of the liver, fatty liver, cholestasis and primary hepatocellular carcinoma are attracting considerable attention. First applied to the measurement of human bile acids in blood plasma in 1965, gas-liquid chromatography has been utilized as a standard method in medical practice. Recently, radioimmunoassay and enzyme linked radioimmunoassay have been utilized. This progress in the estimation of bile acids has awoken our interest in the pathophysiological significance of bile acids in liver diseases. The author reviewed the information on bile acid metabolism in liver diseases which has been reported up to the present. PMID- 6281950 TI - Studies on the mechanism of action of 1 alpha,24-dihydroxyvitamin D3. III. The specific binding of 1 alpha,24-dihydroxyvitamin D3 to the receptor of chick parathyroid gland. AB - Three protein fractions of the cytosol of the chick parathyroid glands, which had the sedimentation constants of 2.5 S, 3.7 S and 5.5 S, were found to bind with 1 alpha,25-dihydroxyvitamin D3. Among these proteins, the 3.7 S protein was assumed to be the specific receptor protein. The 3.7 S receptor protein was also capable of binding to 1 alpha,24-dihydroxyvitamin D3 but not 25-hydroxyvitamin D3. The binding affinity of 1 alpha,24(R)-dihydroxyvitamin D3 to the 3.7 S receptor protein was estimated to be 1.2 times greater than that of 1 alpha,25 dihydroxyvitamin D3, while 1 alpha,25-dihydroxyvitamin D3 bound to the receptor protein about 10 times stronger than 1 alpha,24(S)-dihydroxyvitamin D3. The dissociation constant for the receptor-1 alpha,25-dihydroxyvitamin D3 complex at 0 degrees C was 2.7 x 10(-11) M, the dissociation constants were calculated to be 2.2 x 10(-11) M and 2.6 x 10(-10) M for the complexes with 1 alpha,24(R) dihydroxyvitamin D3 and 1 alpha,24(S)-dihydroxyvitamin D3. PMID- 6281951 TI - Pigeon herpes encephalomyelitis virus in Egypt. AB - A virus was isolated in Egypt from brain and liver collected from domestic pigeons suffering from nervous disorders. The morphological and biophysical properties of the virus were consistent with it being a member of the family Herpetoviridae. Antigenically the virus was closely related to if not identical with pigeon herpes encephalomyelitis virus. PMID- 6281952 TI - [Properties of urease immobilized on the functional organic silica surface]. AB - The paper deals with kinetics of the urea hydrolysis by microbial-origin urease dissolved and immobilized on the organic silica surface. It is shown that hydrolysis kinetics for soluble urease is described by the Michaelis-Menten equation until the concentration of urea reaches 1 M. Two fractions differing in the Michaelis constant are revealed for silochrome immobilized urease. The rate of urea hydrolysis by native and immobilized urease was studied depending on the pH value in presence of the substrate in the 1 M and 5 mM concentration. The hydrolysis rate of 1 M urea in the buffer-free solution by silochrome-immobilized urease is practically independent of pH within 4.5-6.5. Application of a 2.5 mM phosphate-citrate buffer as a solvent causes an increase in the hydrolysis rate within this pH range. For a soluble urease the 1 M urea hydrolysis rate dependence on pH is ordinary at pH 5.8-6.0. If the substrate concentration is 5 mM, the pH-dependences for the rate of the urea hydrolysis by silochrome- and aerosil-immobilized urease are close and at pH above 6.0 coincide with those for a soluble enzyme. The found differences in the properties of soluble and immobilized ureases are explained by the substrate and reaction products diffusion. PMID- 6281954 TI - [New aspects of the metabolism of oxygen. I. Structure of free radicals]. PMID- 6281953 TI - [Influence of thymostimulin on the activity of certain enzymes of adenosine and AMP metabolism in lymphocytes of rats with mammary cancer]. AB - The adenosine deaminase, 5'-nucleotidase, AMP-aminohydrolase and adenylate kinase activities and thymostimulin influence on these indices in vivo were studied in rat thymus and spleen lymphocytes in the latent period of DMBA-induced mammary carcinogenesis. The adenosine deaminase activity was established to increase while 5'-nucleotidase and AMF-aminohydrolase activity to decrease in the thymocytes of intact animals treated with thymostimulin; the adenylae kinase activity of spleen lymphocytes decreased as compared with that in the rats not treated with the preparation. The dynamics of changes in the investigated enzymes in activities in lymphocytes was of wave-like pattern in the latent period. The treatment of animals with thymostimulin in this period normalized adenosine deaminase activity and decreased the activity of the other enzymes in these cells. PMID- 6281955 TI - [Current aspects of oxygen metabolism, II. Function of free radicals]. PMID- 6281956 TI - [Cytomegaloviruses: their role in human pathology]. PMID- 6281957 TI - An investigation into the susceptibility of cattle to bovine leukosis virus following inoculation by various routes. AB - The effect of different routes of inoculation on the incubation period, as indicated by the detection of antibody and by the detection of bovine leukosis virus (BLV) in lymphocytes, were compared. None of the 12-month-old steers exposed to BLV by the oral route developed BLV infection. Intratracheal, subcutaneous and intradermal inoculations were found to be particularly effective in establishing BLV infection, which was detected three to four weeks after inoculation. In the majority of animals, serum antibody and virus were detected at the same time. One out of four in-oestrus heifers inoculated via the uterus with mixtures of BLV infected lymphocytes and semen became infected. It appears that there is an inhibitory factor in fresh semen that prevents BLV infection from becoming established. Viral antigen was detected earlier in BLV infected lymphocytes using the cocultivation method than by electron microscopy to demonstrate BLV particles in mitogen stimulated lymphocytes. PMID- 6281958 TI - Detection of transient and persistent feline leukaemia virus infections. AB - A study was made of cats persistently or transiently viraemic with feline leukaemia virus (FeLV) following experimental oronasal infection. Cats of two ages were exposed to the virus. One group was infected when eight weeks old in the expectation that most of the cats would become persistently viraemic, and the second group when 16 weeks old, so that some would show signs of a transient infection and then recover. The periods following infection when virus was detectable in the blood and in the oropharynx were determined for each group. Three methods for detecting viraemia were compared: virus isolation, immunofluorescence on blood smears and an enzyme-linked immunosorbent assay (ELISA). There was good overall agreement among the three tests in detecting virus-positive cats. Virus was found sooner after infection by virus isolation than by the other methods, and virus appeared in the blood slightly sooner in cats which developed persistent viraemia than in transiently viraemic cats. Infectious FeLV was isolated from the oropharynx of all of the persistently viraemic cats, in most cases simultaneously with virus in the plasma. Virus was also isolated from the mouth of most transiently viraemic cats. Under field conditions such transient excretion of virus lasting only a few days would rarely be detected in a single sampling. This might explain how FeLV is maintained in free range urban cats in the absence of a large number of cats with persistent active FeLV infection. For routine diagnosis, immunofluorescence would appear to offer the best chance of differentiating transient and persistent infections by FeLV. PMID- 6281959 TI - Isolation of a haemagglutinating adenovirus from commercial ducks. PMID- 6281960 TI - A comparison of three methods of feline leukaemia virus diagnosis. AB - Samples of blood from pet cats were examined for evidence of feline leukaemia virus (FeLV) by three techniques: virus isolation, immunofluorescence and an enzyme-linked immunosorbent assay (ELISA) Leukassay F. There was good agreement between the results from virus isolation and immunofluorescence. However, about 30 per cent of cats which were positive for FeLV antigen by ELISA were negative by either of the other tests. The status of most of these cats was unchanged four or 12 weeks later. PMID- 6281962 TI - Runting syndrome in broiler chickens in Australia. PMID- 6281961 TI - Winter dysentery: a coronavirus-like agent in the faeces of beef and dairy cattle with diarrhoea. PMID- 6281963 TI - Alpha-1-antitrypsin in malakoplakia. AB - Macrophages in malakoplakia contain large amounts of immunoreactive alpha-1 antitrypsin (AAT). The amount of AAT remains unchanged during the morphogenetic stages of the pathological process (early, granulomatous, fibrosing phases), and does not correlate with the number or the presence of Michaelis-Gutmann (M.G.) bodies. Macrophages from other pathological processes, closely resembling malakoplakia cells but without M.G. bodies, did not contain AAT, except for a few macrophages in tuberculosis and xanthogranulomatous pyelonephritis. Whatever the source and the pathogenic role of AAT in malakoplakia, its presence in all macrophages seems to be specific for this disease. Immunohistochemical staining for AAT is therefore proposed as a useful test for an early and accurate differential diagnosis of malakoplakia. PMID- 6281964 TI - Angiomatoid malignant fibrous histiocytoma. Case report and electron microscopic findings. PMID- 6281965 TI - Mouse mammary tumor virus DNA sequences in tumorigenic and nontumorigenic cells from a mammary adenocarcinoma. PMID- 6281967 TI - Shifts in the extent and patterns of DNA methylation upon explanation and subcultivation of adenovirus type 12-induced hamster tumor cells. PMID- 6281966 TI - A solid-phase assay of solubilized HeLa cell membrane receptors for binding group B coxsackieviruses and polioviruses. PMID- 6281969 TI - Monoclonal antibodies to xenotropic and MCF murine leukemia viruses derived during the graft-versus-host reaction. PMID- 6281968 TI - Sendai virus DI RNA species with conserved virus genome termini and extensive internal deletions. PMID- 6281970 TI - Evidence against involvement of host transcription in the replication of vaccinia and herpes simplex viruses. PMID- 6281971 TI - Persistent infection by equine infectious anemia virus: asymmetry of nucleotide sequence reiteration in the integrated provirus of persistently infected cells. PMID- 6281972 TI - Molecular cloning and partial nucleotide sequence of human papillomavirus type 1a DNA. PMID- 6281973 TI - Sendai virus gene sequences identified by oligonucleotide mapping. PMID- 6281975 TI - Isolation of transformation-defective host-range mutants of polyoma virus on normal mouse cells. PMID- 6281974 TI - Restriction mapping and physical characterization of the cottontail rabbit papillomavirus genome in transplantable VX2 and VX7 domestic rabbit carcinomas. PMID- 6281976 TI - Cell tropism and expression of mouse hepatitis viruses (MHV) in mouse spinal cord cultures. PMID- 6281977 TI - The white pock mutants of rabbit poxvirus. IV. The late white pock (mu) host range (hr) mutants of rabbit poxvirus are blocked in morphogenesis. PMID- 6281978 TI - Loss of different pericellular matrix components of rat cells transformed with a T-class ts mutant of Rous sarcoma virus. PMID- 6281979 TI - Monoclonal antibodies to murine hepatitis virus-4 (strain JHM) define the viral glycoprotein responsible for attachment and cell--cell fusion. PMID- 6281980 TI - Two retroviruses with similar transforming genes exhibit differences in transforming potential. PMID- 6281981 TI - Analysis of epstein-Barr virus (EBV) of P3HR-1: isolation of EBV with EBNA induction ability in human cord lymphocytes and without EA induction ability in Raji cells. PMID- 6281982 TI - Visna DNA synthesis and the tempo of infection in vitro. PMID- 6281983 TI - Avian lymphoid leukosis virus infection and DNA integration in the preleukotic bursal tissues: a comparative study of susceptible and resistant lines. PMID- 6281984 TI - Plasmid-bacteriophage recombination. II. Density transfer analysis. PMID- 6281985 TI - Effect of tunicamycin on the replication of Sendai virus. PMID- 6281986 TI - Identification of plaque isolates of an avian retrovirus causing rapid and slow onset osteopetrosis. PMID- 6281988 TI - [Features of the regulation of key enzymes involved in glycolysis and the pentosephosphate shunt in tissues with different functional specializations]. AB - Under conditions of acute and chronic stress carbohydrate metabolism was decreased in muscles and liver tissue, increased in adrenal glands and unaltered in brain. cAMP at concentration 10(-5)M decreased the rate of glycolysis in liver tissue and increased it slightly in adrenal glands. The inhibitory effect of cAMP was not decreased by 2',3'-AMP and was reversed completely by the protein inhibitor of proteinkinase. cAMP inhibited hexokinase (HK) isoenzymes 2 and 3 as well as glucose-6-phosphate dehydrogenase (G6PD) isoenzymes 1 and 2, activities of HK 1 and G6PD 3 and 4 were unaltered. The decrease in activities of HK and G6PD appears to be related to phosphorylation of proteinkinase; lack of the effect of cAMP in adrenal glands was apparently due to absence of corresponding cAMP-dependent proteinkinase in this tissue. PMID- 6281987 TI - [Prevention of structural damage to myocardial DNA caused by emotionally painful stress by means of beta-adrenoreceptor and lipid peroxidation blockade]. AB - Emotional-painful stress was accompanied by an increase in the rates of depolymerization and of repairatory replication of heart muscle DNA. When the repairatory replication of DNA was repressed by actinomycin D, the rate of its depolymerization was simultaneously increased. A beta-adrenoreceptor blocking agent inderal and an inhibitor of lipoperoxidation ionol prevented the DNA depolymerization and activation of the polymer synthesis under the stressory conditions. These data corroborate the hypothesis that stress favors an increase in content of catecholamines, increasing the rate of lipoperoxidation and producing free radicals responsible for impairment of heart muscle DNA. In ontogenesis the multiple damage-repair cycles in DNA apparently lead to accumulation of genetic errors, which may be among the reasons of senescense. PMID- 6281989 TI - [Cerebral gangliosides in dogs in experimental allergic encephalomyelitis]. AB - Content of gangliosides was decreased in white substance of brain hemispheres and in spinal cord of dogs with typical symptoms of allergic encephalomyelitis. The decrease in content of less polar fractions was compensated by an increase in concentration of more polar gangliosides. After an intracisternal administration of the antibrain antibodies the content of gangliosides was also decreased in brain cortex and white substance of hemispheres. The data obtained suggest that the neurone membranes are impaired under the pathological conditions as well as that the antibrain antibodies are important for development of the structural impairments observed. PMID- 6281990 TI - [Klippel-Trenaunay syndrome]. PMID- 6281991 TI - [Herpes simplex]. AB - Publications concerning herpes virus hominis and other viruses of the herpes group have appeared with increasing frequency in the last few years. The mechanism of recurrence is yet not fully elucidated and therapy still remains a problem. The fulminant course of herpes sepsis in the newborn and the generalized herpes infection in the immunodeficient patient can be treated only with difficulty. Neurologic and ophthalmic disease in children can often be linked to neonatal herpes infections. In the category of sexually transmitted diseases, herpes genitalis is becoming more significant. Numerous observations indicate that herpes virus hominis and other members of the herpes group are potential oncogenic agents. This paper presents an overview of recent findings. PMID- 6281992 TI - [Herpes zoster]. AB - This paper summarizes present knowledge on pathogenesis and therapy of herpes zoster. Formerly, zoster was considered a reinfection of a partly immune individual by the varicella-zoster virus localized into a body segment of lowered resistance. Recent epidemiological and virological evidence, however, suggests a different mode of pathogenesis: in the course of varicella, VZV migrates via the sensory nerves to the dorsal root ganglia where it remains in an inactive form. It may be reactivated in response to a local or systemic lowering of the level of host resistance, resulting in painful ganglionitis and, by descension, in the characteristic segmental skin lesions of herpes zoster. Treatment with systemic corticosteroids in the early stages of zoster helps to decrease the frequency and severity of postzosteric neuralgias. Corticosteroids ought to be given at a relatively high dosage and be tapered only slowly in the course of a few weeks. They are particularly useful in older individuals, who constitute a population prone to postzosteric neuralgias. In severe cases of generalized zoster, corticosteroids should not be given to avoid further lowering of host resistance. Recently, antiviral drugs have been developed which combine high effectiveness with very tolerable side effects, namely vidarabine and acyclovir. PMID- 6281993 TI - [Condylomata acuminata]. AB - Condylomata acuminata are benign epitheliomas which are caused by the human papilloma virus. 8 subgroups (variants) of the virus are known up to now. Condylomata acuminata are infectious, the virus being transferred by direct contact with a lesion. Condylomata acuminata are found more often in males than in females and never occur in children prior to the age of two years. The peak incidence is between 19 and 22 years of age. Greatly proliferating condylomata acuminata (type Buschke-Lowenstein) may lead to malignant degeneration. There exists a broad spectrum of topical therapeutic modalities: cytotoxic substances such as podophyllin (10 to 25%), colchicine (8%), 5-fluorouracil (1 to 5%), cryotherapy (solid carbon dioxyde or liquid nitrogen), surgical treatment (excochleation, electrocautery or laser beam. Vaccination therapy may prove successful in certain cases in the future. PMID- 6281994 TI - [Laboratory diagnosis of viral diseases of the skin (author's transl)]. AB - In order to establish the diagnosis of viral diseases with clinical manifestations in the skin, frequently the help of a virus laboratory is needed. Not even classical childhood diseases such as measles and rubella can safely be diagnosed by the clinician. Today, in many cases diagnosis is possible within hours after the arrival of clinical specimens in the laboratory. Only in cases of enterovirus infection and tropical diseases has the virologist to resort to the classical time-consuming methods of direct (virus isolation) or indirect (serologic) diagnostic virology. PMID- 6281995 TI - [Immunohistochemical studies on the association between hepatitis B surface antigen and primary hepatocellular carcinoma (author's transl)]. AB - In many geographical areas especially in African and South-east-Asian countries hepatitis B virus infection is considered to be a major etiological factor in the development of primary hepatocellular carcinoma. 107 autopsy- and 15 biopsy specimens were studied by means of immunohistochemistry [peroxidase antiperoxidase-(PAP-)method] to demonstrate the association between primary hepatocellular carcinoma and hepatitis B surface antigen (HBsAg). HBsAg was found in 8 of 107 tumour specimens (7.4%) and liver cirrhosis in 102 of the 107 autopsy specimens with hepatocellular carcinoma (95%). 10 of the 15 biopsy-specimens showed neoplastic and non-neoplastic liver tissue, and in 2 of these 10 cases HBsAg was found. Liver cirrhosis could be seen in 9 of those 10 specimens. HBsAg was also studied in 90 cases with liver cirrhosis and was found to be positive in 2 of them (2,2%). HBsAg associated with primary hepatocellular carcinoma was only found in non-neoplastic liver cells of cirrhotic livers. Our studies indicate that in our geographical area the association of HBsAg with primary hepatocellular carcinoma is much less conspicuous than in Asian, African and even Southern European communities. PMID- 6281996 TI - The use of on-line high-performance liquid chromatography-mass spectrometry for the identification of ranitidine and its metabolites in urine. AB - 1. Reverse and normal phase h.p.l.c. systems have been developed for separation ranitidine and three metabolites, desmethylranitidine, ranitidine-S-oxide and ranitidine-N-oxide. 2. These h.p.l.c. systems have been evaluated for characterization of ranitidine and metabolites using an h.p.l.c. coupled to a mass spectrometer with a moving belt interface. 3. Ranitidine and its metabolites were thermally degraded under the conditions required to evaporate the reverse phase eluent that contained 40% aq. 0.05 M ammonium acetate. 4. The normal phase eluent was evaporated in the interface at a lower temp. and satisfactory mass spectra were obtained from 1 microgram of ranitidine and each metabolite injected on to the h.p.l.c. column. 5. Normal phase h.p.l.c.-mass spectrometry has been used to identify ranitidine and three of its metabolites in rabbit and human urine obtained after oral administration of ranitidine. PMID- 6281997 TI - [Surgery of benign pancreatic diseases]. PMID- 6281998 TI - [The problem of sequential cancers after antineoplastic chemotherapy or radiotherapy]. AB - It is reported on 4 patients who in a period of 2 to 14 years after successful therapy of a malignant tumour using the therapy with cytostatics and (or) radiotherapy fell ill with a genuine secondary neoplasia. A connection of the therapeutic procedures mentioned with the repeated tumour induction is obvious, a genetic disposition (cancer diseases in parents and brothers and sisters) might favourably cooperate in three of the cases mentioned. It is referred to the necessity of strongest indication to the therapy with cytostatics and ionising rays. PMID- 6281999 TI - [Disorders of electrolyte balance and metabolism in hormone-secreting bronchial cancer]. AB - Four cases of bronchial carcinoma were observed which showed the picture of an ectopic ACTH-hormone syndrome. References to an endocrinopathy resulted from the hypopotassaemia and the metabolic alkalosis. Signs of Cushing's syndrome appeared only in one case. By disturbances of the electrolyte and acid-base-balance the functions of the skeletal and smooth musculature as well as of the heart muscle were much impaired. In two cases the clinical symptoms and the laboratory findings still before the manifestation of the bronchial carcinoma, in the third case simultaneously referred to the correct diagnosis. Due to the misleading cardiac findings and the fulminant course of the disease in the fourth case the diagnosis was clarified only by autopsy. the disturbances of the ion balance and of the metabolism had contributed to the respiratory insufficiency, had evoked disturbances of the cardiac rhythm and had unfavourably effected on the course of of the disease. PMID- 6282000 TI - Plasma digoxin concentrations during administration of dietary fibre (guar gum) in man. AB - The effect of guar gum on digoxin absorption was investigated in eleven healthy volunteers. The drug was administered in the morning during two five-day-periods, together with a liquid mixed test meal (+/- 18 g guar). By measuring plasma digoxin concentrations no differences were registered between the control and guar gum period. It is concluded that long-term administration of guar gum will not interfere with adequate digitalization (digoxin bioavailability). PMID- 6282001 TI - Activities of cytoplasmic, mitochondrial and brush border enzymes in jejunal mucosa of chronic alcoholics. AB - The activity of certain enzymes of the energy producing metabolism of the cytoplasmic and mitochondrial compartment and of disaccharidases was determined in jejunal biopsies of 24 chronic alcoholics (CA) and 10 non-alcoholic control subjects (C). The activity of glucokinase, an enzyme of glycolysis, was markedly (44%, p less than 0.05) increased in the biopsies obtained from CA, while the activity of fructose bisphosphatase, an enzyme of gluconeogenesis, was significantly (p less than 0.05) depressed in CA when compared to C. The activity of other glycolytic enzymes was not affected in CA. The activity of L-alanine amino-transferase was lower in CA (p less than 0.05). A reduction was also seen for mean succinate dehydrogenase activity in CA (-30%), however, this difference was not statistically significant. The mean activity of lactase, maltase and sucrase was comparable in both groups. PMID- 6282002 TI - [Subungual tumours (author's transl)]. AB - The differential diagnosis of subungual tumours is often very difficult, since the same tumor located in another area looks entirely different. Former infections or traumata may further mask the tumour. Pain is usually not characteristic. Infection and the history of a trauma may further mask this tumour. Pain is usually not characteristic. both malignant and benign tumours may develop from all tissues present in the distal phalanx. Subungual melanomas are the most common malignant subungual tumours, whereas a variety of benign tumours occurs in the subungual region, the most characteristic of them is the glomus tumour. PMID- 6282003 TI - Collagen peptidase in chronic liver disorders. PMID- 6282004 TI - [Therapy of drug addicts]. PMID- 6282005 TI - [Systems integration of pain excitation]. PMID- 6282006 TI - Gene frequencies of red-cell uridine-5-monophosphate kinase (UMPK) in Western Germany (Dusseldorf region). PMID- 6282007 TI - [Histological, morphometric and topochemical analysis of placentas from pregnancies with EPH Gestosis (author's transl)]. AB - The authors' studies have shown three pathological phenomena to be predominant in placentas of women with EPH gestosis: disappearance of acid mucosubstance from wall of spiral arteries, reduction of ATPase, 5-Nase, and aP activities on apical and basal surfaces of terminal villi, and numerical increase as well as de differentiation of trophoblastic nodes. Vulnerability of vascular walls to the action of vasodepressive factors is aggravated, according to the authors, in cases of complicated gestosis in which aMPS levels in the muscular membranes of spiral arteries are inadequate. PMID- 6282008 TI - [Foetal complications on extremities by amniotic bands--case report (author's transl)]. AB - Reported in this paper are malformations of foetal extremities due to ligatures caused by amniotic bands. These were accompanied by syndactylism. Causes and therapeutic approaches are discussed. PMID- 6282009 TI - Effects of vitamins A, D3, E, and C on the characteristics of bull semen. PMID- 6282010 TI - [Standardization of an indirect enzyme-linked immunosorbent assay (ELISA) for demonstration of antibodies to chicken bronchitis virus]. PMID- 6282011 TI - A bovine thymic lymphosarcoma case showing a negative serological response to bovine leukemia virus antigens, in a herd with high incidence of enzootic bovine leukosis. PMID- 6282012 TI - [Epidemiology of infectious mononucleosis]. PMID- 6282013 TI - [Interrelation of the toxicity and capacity of pertussis vaccines to alter the body immune response to heterogenetic antigens]. AB - The influence of immunization with pertussis vaccines differing in toxicity on the intensity of the formation of antibodies to heterologous antigens (S. typhi Vi-antigen) and on the resistance of the body to natural infection (S. typhimurium) was studied in mice. The toxicity of pertussis vaccines was found to be related to their capacity for changing immune response to heterologous antigen. In mice showing pronounced toxicosis the injection of pertussis vaccine resulted in a decrease in their capacity for Vi-hemagglutinin formation. The appearance of a definite degree of resistance ot S. typhimurium was observed in mice previously immunized with pertussis vaccine possessing pronounced toxic properties. Nevertheless, the appearance of enhanced resistance to infection was observed only in the animals previously immunized with a nontoxic preparation. PMID- 6282014 TI - [Carbon assimilation from a NaHCO3 nutrient medium by an STI-1 strain culture]. PMID- 6282015 TI - [Analysis of the results of serum studies of children and adults for antibodies to the poliomyelitis virus in a multiyear effort at vaccinal prevention of this infection]. AB - The results of testing the blood sera of children and adults for the presence of antibodies to poliomyelitis have shown the low level of immunity to this infection. The authors believe that the main reasons of the tendency towards the decrease in immunity to poliomyelitis, observed in recent years, are the drawbacks of the vaccinal prevention of this infection and the absence of a differentiated approach to the choice of immunization methods under concrete conditions. Mass immunization against poliomyelitis is recommended, especially in the southern and south-eastern regions of the U.S.S.R. PMID- 6282016 TI - [Dry medium for isolating Clostridium perfringens]. AB - The possibility of using blood substitutes (hydrolysin, casein hydrolysate "tsolipk" and amino peptide) with expired shelf life as a culture medium for the isolation of Cl. perfringens has been studied. Dry culture medium based on these inedible products has been developed. To stimulate bacterial growth, fodder yeast extract has been used. The suppression of the growth of extraneous microflora is ensured by the use of antibiotics: polymyxin sulfate and mycerin sulfate. The recommended medium is not inferior in its quality (sensitivity, rapidity of growth, differentiating and inhibiting properties, etc.) to media based on meat and casein. The use of the newly developed medium is economically grounded and allows one to obtain standard results. PMID- 6282017 TI - [Standardization of preparations intended for the serodiagnosis of arboviruses]. AB - As the result of our research work, 3 reference preparations have been first obtained and studied in accordance with all requirements of biological standardization. These preparations are the national standard of yellow fever antiserum and immune ascitic fluids (IAF) used as reference reagents: IAF to tick borne encephalitis virus and IAF to Japanese encephalitis virus. The new preparations are stable, possess sufficient specific activity and can be used as standard preparations for the identification of the above-mentioned viruses. PMID- 6282018 TI - [Basic principles of health resort treatment of children with sequelae in nervous system diseases]. AB - As a result of many-year studies of the effectiveness of using health resort factors for treating children with various nervous system diseases basic principles of that treatment have been determined. A particular importance is attached to combined application of different health resort factors with consideration for the specificity of their therapeutic action and the peculiarities of the disease. The scope of the therapeutic measures should be dynamic in character and dose, so as to preserve the effect of each principal therapeutic factor on the patient's body. Attention is paid to the necessity of careful selection of patients for the health resort treatment, as well as to he therapeutic strategy in subsequent periods. Data on the effectiveness of treating various groups of patients at health resorts are presented. PMID- 6282019 TI - The turnover of liver cytosol proteins in very old rats. AB - The protein degradation rate of total liver homogenate as well as of the liver cytosol was measured according to the method introduced by GARLICK and coworkers (Biochem. J. 156 (1976) 657-663). The half-lives were 2.85 days (cytosol) and 2.45 days (homogenate) in the group consisting of 29 months old rats and 2.35 days and 2.15 days in the control group consisting of 12 months old rats. The amount of cathepsin D determined in some organs by two independent methods (immunological and enzymic) revealed differences in the group of the old animals. We suggest the accumulation of cross reacting material of cathepsin D in rats of the old group. It is perhaps one of the reasons why a slower protein degradation is observed in old animals. PMID- 6282020 TI - Regulation of intracellular protein degradation by insulin and growth factors. AB - Protein breakdown in many cell lines is inhibited by growth factors. The response is specific, rapid and additive only at suboptimal concentrations. Transformed cells are typically more sensitive to growth factors than nontransformed cells while the effectiveness of growth factors in human fibroblasts is diminished with senescence. Bovine colostrum has been shown to be an extremely rich source of growth factors as compared to serum. PMID- 6282021 TI - Globular protein stability: aspects of interest in protein turnover. AB - The conformational stability of globular proteins is remarkably low. Under physiological conditions, the native globular conformation is only from 5 to 15 kcal/mole more stable than unfolded conformations. In addition, small changes in the structure of a protein such as removing one terminal residue or cleaving a single peptide bond frequently lead to a substantial decrease in the stability. Likewise, single substitutions in the amino acid sequence can increase or decrease the stability by several kilocalories per mole. The low conformational stability of globular proteins and the sensitivity to small changes in structure suggest a possible role for conformational stability in the intracellular degradation of proteins. Several lines of evidence from in vivo studies of protein degradation are consistent with this idea. PMID- 6282023 TI - Stabilizing effects by Mg2+ on Na,K-ATPase. PMID- 6282022 TI - On the substrate specificity of cathepsin L. AB - A view is given on the maximal hydrolysis of proteins by cathepsin L (EC 3.4.22.15) in dependence on the pH. The overall degradation of several proteins at pH values lower than pH 6.0 implies a very broad specificity, whereas at pH 7.0 and 7.5 cathepsin L seems to act on proteins cleaving only restricted specific peptide bonds. Some kinetic constants are given for the three synthetic substrates of cathepsin L which are known so far: Bz-Arg-NH2, Z-Lys-OPhNO2 and Z Phe-Arg-NMec. They cannot be used as completely specific substrates of cathepsin L, because all of them are hydrolysed by cathepsin B and also other proteinases. PMID- 6282024 TI - Fluctuations in the plasma angiotensin I converting enzyme activity during long term treatment with captopril. AB - Eight hypertensive patients on chronic captopril treatment were studied: blood pressure, plasma converting enzyme activity (pCEA) and various components of plasma renin-angiotensin-aldosterone system were measured repeatedly, immediately before and up to 7 hours after the usual morning dose of captopril in 5 patients, or a matching placebo in 3 patients. In the patients, receiving a placebo no significant changes were observed over a 7 hour period in pCEA, plasma renin activity (PRA), plasma angiotensin II (ANG II) plasma aldosterone (PAC) and blood pressure. In patients receive captopril (200 mg) pCEA rapidly decreased, reaching after 2 hours a minimum, corresponding to nearly 20 per cent of its reference value. Thereafter pCEA increased and after 7 hours remained only slightly depressed. Within the first hour after captopril intake a small but significant decrease of ANG II and PAC was observed, while PRA and blood pressure remained unchanged throughout the study period. A continuous low pCEA level is therefore not necessary to achieve a sustained blood pressure lowering effect during chronic captopril treatment. PMID- 6282025 TI - Cytologic features of nonfatal herpesvirus tracheobronchitis. AB - Herpesvirus infections of the lower respiratory tract have most commonly been reported in patients with severe burns, immunosuppression or malignancies. Two patients without any of these underlying conditions developed severe herpetic tracheobronchitis, diagnosed by cytologic examination and confirmed by serologic studies. Serial examination of sputum, bronchial brushings and bronchial washings permitted observation of the evolution and progression of cellular changes found in herpesvirus infection of the lower respiratory tract. both patients recovered without specific antiviral therapy, but both developed superinfection with gram negative organisms, requiring intensive antibiotic therapy. The distinctive features of herpesvirus infection in the tracheobronchial tree are similar to those recognized elsewhere in the body. Early findings include a variety of nonspecific changes in nuclear chromatin configurations; multinucleated cells may be common but do not often contain the central intranuclear inclusion bodies seen in later stages. These distinctive central intranuclear inclusions disappear in a few days, leaving only reparative changes in the surface epithelium. Herpesviruses are increasingly being reported in the literature as an etiologic agent of acute tracheobronchitis in otherwise healthy individuals. PMID- 6282026 TI - Hormones in the seminal fluid. The transport proteins of the thyroid hormones. AB - The study of the retention of titrated T3 on columns of Sephadex G-25, (Resin T3 Uptake), in the presence of untreated plasma, or plasma treated with charcoal, indicates that there are very small quantities of protein-hormone compounds present in the seminal fluid. In cases of phlogosis of the genital tract, the increase of albumin in the seminal fluid might determine an increase in the binding capacity with titrated T3. This would therefore give the possibility of a further test in the diagnostics of male infertility. PMID- 6282027 TI - Serum beta-ELI circulating levels and primary amenorrhea. PMID- 6282028 TI - Partial myeloperoxidase deficiency. AB - Neutrophil myeloperoxidase (MPO) activity was analyzed by a semi-quantitative cytochemical method in 268 subjects divided into several groups. 17 subjects with significantly reduced MPO activity were found: 11 of 23 in the preleukemia group, 2/14 AMLs, 1/20 myeloproliferative syndrome, 1/7 carcinoma with bone marrow metastases, 1/33 diabetes mellitus and 1/50 normals. Only in the preleukemia group, was MPO significantly reduced in comparison to the normal group (p less than 0.005). The high frequency of acquired MPO deficiency in preleukemia represents a useful criterium for this diagnosis. Furthermore, in these patients, as well as in the other subjects studied, no apparent correlation between MPO level and infection could be demonstrated. PMID- 6282029 TI - [Morphological studies of bone marrow neutrophilic and acidophilic granulocytes of white rate during stimulated leukopoiesis]. AB - The neutrophils and eosinophils of rat bone marrow were studied after stimulating bone marrow with leucogen administered in therapeutic doses. The morphological examinations of these cells were carried out under electron microscope analysing the activity of myeloperoxidase. Disturbances were observed in the formation of primary and secondary granules in the neutrophils. This suggest a decrease of their functional efficiency. The formation of granules in the neutrophils was similar to normal. After 7 days of leucogen administration the stability between the number of granules with core and without it was disturbed in favour of the latter ones. After 14 days the proportions in the number of both types of granules reverted to the normal values. PMID- 6282031 TI - Cyclic nucleotides in cerebrospinal fluid of patients with intracranial and spinal tumors. AB - Since the cyclic nucleotides (CN) adenosine 3',5'-monophosphate (cAMP) and guanoside 3',5'-monophosphate (cGMP) are involved in the regulation of cell proliferation and tumor growth in vitro, a study was made of the levels of these compounds in the lumbar cerebrospinal fluid (CSF) of 20 patients with intracranial and spinal tumors. In the presence of benign or malignant intracranial tumors there was a slight and not significant decrease of cAMP as well as cGMP levels in the CSF, as compared to control patients. While there was no significant correlation between the levels of the two CN in controls, there was a positive correlation in tumor patients. Total protein content and cAMP were negatively correlated in malignant intracranial tumors. Possible influences of tumor growth and intracranial pressure increases on CN levels are discussed. In spinal tumor patients normal CN levels were observed. However, in a patient with meningeal sarcoma an extremely marked elevation of cAMP occurred in parallel with the extension of the tumor to the spinal meningeal space, suggesting massive secretion of cAMP from the tumor cells. PMID- 6282030 TI - Towards a specific histochemical localization of adenylate cyclase in the rat hippocampus. I. Methodical aspects. AB - The adequate histochemical demonstration of adenylate cyclase still involves serious methodical problems (low basal activity, fixation procedures, heavy metal ions as capture agents etc.). We have tried to revise fixation approach and composition of incubation medium in order to specify the mode of demonstration of the enzyme. The influence of strontium and barium (instead of lead) as well as dithiothreitole and sodium fluoride on the survive of adenylate cyclase activity was studied using a radiochemical model system. PMID- 6282032 TI - Adrenergic and peptidergic neuromuscular mechanisms in the human Fallopian tube, with special regard to cyclic influences. PMID- 6282034 TI - Studies on humoral immunity and HLA antigens in multiple sclerosis, optic neuritis and hereditary optic atrophy. PMID- 6282033 TI - Estrogen replacement therapy after the menopause. Estrogenicity and metabolic effects. AB - Certain metabolic effects were investigated in post-menopausal women undergoing oral estrogen replacement therapy for 6 months using various substances. The increases in serum concentration of the estrogen-sensitive proteins, pregnancy zone protein (PZP), and sex hormone binding globulin (SHBG) had very similar and dose-dependent patterns. Ethinyl-estradiol was found to be much more potent than the "natural" estrogens. Estriol in various doses did not increase the protein level. Gonadotropin inhibition occurred in a dose-dependent manner. In terms of FSH suppression ethinyl-estradiol was approximately 120 times as potent as the "natural" estrogens. There was a striking resemblance between the "estrogenicity" of four different estrogens when expressed both in inhibition of gonadotropins and in induction of the two serum proteins SHBG and ceruloplasmin. Estriol caused a significant depression of FSH when given orally in a dose of 2 mg three times daily. Prolactin was found to decrease during treatment with low doses of estrogens. Estrogen therapy was found to have only moderate effects on adrenal androgens. Tamoxifen, and anti-estrogen, was found to exert distinctly estrogenic effects during treatment of post-menopausal women. In post-menopausal women with low amounts of circulating estrogens the tamoxifen-receptor complex itself may produce a net estrogenic response. Serum samples from post-menopausal women treated with ethinyl estradiol 0.05 mg and estrone sulphate 2.5 mg daily were found to reduce the lymphocyte reactivity in mixed lymphocyte cultures. PMID- 6282035 TI - The Kerala Decentration Meter. A new method and devise for fitting the optical of spectacle lenses in the visual axis. AB - Centring of spectacle lenses is much neglected field of ophthalmology. The prismatic effect caused by wrong centring results in a phoria on the eye muscles which in turn causes persistent eyestrain. The theory of visual axis, optical axis and angle alpha is discussed. Using new methods the visual axis and optical axis of 35 subjects were measured. The results were computed for facial asymmetry, parallax error, angle alpha and also decentration for near vision. The results show that decentration is required on account of each of these factors. Considerable correction is needed in the vertical direction, a fact much neglected nowadays; and vertical decentration results in vertical phoria which is more symptomatic than horizontal phorias. Angle Alpha was computed for each of these patients. A new devise called 'The Kerala Decentration Meter' using the pinhole method for measuring the degree of decentration from the datum centre of the frame, and capable of correcting all the factors described above, is shown with diagrams. PMID- 6282036 TI - DNA measurements of human pituitary tumours. PMID- 6282037 TI - [Pathogenesis and therapy of cervical tracheal stenoses]. PMID- 6282038 TI - Physiological and pathogenic effects of sound. PMID- 6282039 TI - The inner ear and hyperbaric conditions. An electrophysiological and morphological study. PMID- 6282041 TI - Effect of glucagon infusion on blood glucose, plasma immunoreactive insulin, growth hormone and adenosine 3'5'-monophosphate in obese children. AB - The behaviour of plasma glucose, immunoreactive insulin, growth hormone and adenosine 3',5'-monophosphate (cAMP) in response to glucagon infusion was investigated in obese and control children. Hyperinsulinaemia and low growth hormone levels were found in the obese group. Hyperinsulinaemia did not prevent the glucagon-induced cAMP release in obese children. The plasma cAMP concentration decreased after the first hour of glucagon infusion in both groups. The decline of cAMP concentration was more rapid in the obese group than in the controls, resulting in a significantly lower plasma cAMP level at the end of the glucagon load. PMID- 6282040 TI - Aminoglycoside-induced cochlear pathology in man. AB - Temporal bones from five patients with hearing loss as a result of aminoglycoside treatment were examined by the method of microdissection and surface preparations, followed by celloidin embedding and serial sectioning of the modiolus. Three patients had received the newer antibiotics, gentamicin, tobramycin, and amikacin; the other two neomycin. In the cochleas from two patients of the first group there was only a small loss of hair cells, restricted to the lower end of the basal turn. The third, who had been treated with several antibiotics over a longer period of time, showed more extensive but strikingly asymmetrical patterns of degeneration in the two ears. This patient, as well as the fourth, who had received neomycin during peritoneal lavage, had numerous patchy areas of complete disappearance of Corti's organ in the basal turn, with incipient degeneration of the distal ends of the nerve fibers in adjacent portions of the osseous spiral lamina. The fifth patient, who had become deaf after prolonged treatment with neomycin by mouth, showed a complete loss of cochlear hair cells. Nerve fibers were present only in the middle and upper turns, where supporting cells remained. Midmodiolar sections showed a proportionately much greater loss of the distal than of the proximal processes of the cells of the spiral ganglion. These findings underscore once again the special hazard for the inner ear that is associated with the clinical use of neomycin, regardless of the route of administration. PMID- 6282042 TI - Lamellar bodies in benign and malignant schwannomas. AB - A light and electron microscopic study was done on seven cases of schwannomas, four of which were benign and three were malignant. Spherical or oval cytoplasmic inclusion bodies, one to three microns in diameter, were detected in all cases. These inclusions were demonstrable by Luxol fast blue stain. Ultrastructurally, the inclusions appeared as whorls of cell membranes reminiscent of myelin sheath. These lamellar bodies probably reflect the peculiar membrane characteristics of Schwann cells, and they provide a useful marker for recognition of benign or malignant schwannomas. PMID- 6282043 TI - [The anti-stress effect of saponins extracted from panax ginseng fruit and the hypophyseal-adrenal system (author's transl)]. PMID- 6282044 TI - Mechanisms of beta-adrenergic desensitization in rat myometrium. AB - This study was performed in order to elucidate the mechanism behind the decreased responsiveness to beta-adrenergic stimulation occurring in uterine muscle after prolonged treatment with isoprenaline. Pretreatment of rats with isoprenaline, 20 nmol/kg, three times daily during four days, significantly decreased the myometrial relaxing effect of the beta-agonist. There was also a significant decrease of the beta-receptor binding capacity of the myometrial membranes measured by the (--)-(3H) DHA binding technique. In the animals pretreated with isoprenaline no significant increase of the adenylate cyclase activity could be observed after isoprenaline stimulation in vitro. The uterine cAMP level was diminished in the desensitized rats. The phosphodiesterase activity was increased. Thus both decreased production and increased degradation contribute to the lower level of uterine cAMP content. The activity of cAMP dependent protein kinase was also depressed. In this work, where low concentrations of isoprenaline have been administered in vivo, several biochemical parameters have been shown to contribute to the beta-adrenergic desensitization in myometrial tissue. PMID- 6282045 TI - Correlation between vascular smooth muscle relaxation and increase in cyclic GMP induced by some organic nitro esters. AB - Three different nitro compounds glyceryl mononitrate (GMN), glyceryl dinitrate (GDN) and ethylene glycol dinitrate (EGDN) were tested on histamine contracted bovine mesenteric artery. GDN and EGDN caused a dose-dependent relaxation which was accompanied by an increase in the endogenous cGMP level. The ED50-value for GDN and EGDN with regard to the relaxant action was 1.06 X 10(-6) M and 4.20 X 10(-8), respectively. GMN was almost completely ineffective as a relaxing agent and did not cause any significant change in cGMP. Regression analysis revealed a significant correlation between relaxation and increase in cGMP for both GDN and EGDN. The regression model for EGDN could be significantly improved by including the squared cGMP change, indicating a non-linear relationship. With regard to GDN a hyperbolic relationship between relaxation and cGMP increase was found. A slight improvement of the regression model was found for EGDN when the change in cAMP was included. For GMN and GDN no improvement of the regression model could be revealed by including the change of cAMP. It is suggested that the present data give further evidence for cGMP as a mediator of vascular smooth muscle relaxation induced by nitro compounds. PMID- 6282046 TI - Effect of furosemide on parathyroid hormone stimulated guinea pig renal adenylate cyclase and thyrotrophin and fluoride stimulated human thyroid adenylate cyclase. AB - The effect of furosemide 8 X 10(-4) mol/l an 8 X 10(-5) mol/l on parathyroid hormone stimulated adenylate cyclase was studied in renal tissue slices from guinea pigs. Furosemide caused a dose-dependent inhibition of the effect of parathyroid hormone on production of cyclic AMP, without having any significant effect on the basal cyclic AMP production. Furosemide in similar concentrations did not inhibit the stimulatory effect of thyrotrophin and fluoride in human thyroid homogenates suggesting that furosemide is not an universal inhibitor of adenylate cyclase and that the inhibition is not caused by a direct action of furosemide on the adenylate cyclase enzyme. Furosemide did not interfere with binding of cyclic AMP to cyclic AMP binding protein kinase from rabbit muscle. The results indicate that furosemide exerts an inhibitory influence either upon binding of parathyroid hormone to renal receptors or upon transmission of impulse from receptor to adenylate cyclase. The inhibitory influence of furosemide on parathyroid hormone action in kidney could explain the value of furosemide in the acute treatment of hypercalcaemia, but also suggest that chronic treatment with furosemide might interfere with normal calcium metabolism. PMID- 6282047 TI - Receptor development and hormone actions. Effect of insulin and epinephrine on the glycogen content of newborn rat liver cultures. AB - In the 9-day cultures of liver cells isolated from newborn rats epinephrine was found to increase the number of glycogen containing cells, and caused an elevation of the glycogen content of individual liver cells. Insulin treatment had a weak effect on the number of glycogen containing cells and its effect on intracellular glycogen content was negligible. The experiments indicate that the epinephrine receptor develops in the fetal period and is reactive in the newborn animal, while the insulin receptor is not. It was remarkable that in this period of life and under in vitro conditions the effect on cellular glycogen deposition of epinephrine and insulin is similar. PMID- 6282048 TI - Effects of intraventricular administration of cholecystokinin octapeptide sulfate ester and unsulfated cholecystokinin octapeptide on active avoidance and conditioned feeding behaviour of rats. AB - The effects of cholecystokinin octapeptide sulfate ester (CCK-8-SE) and unsulfated cholecystokinin (CCK-8-NS) were studied following intraventricular administration on active avoidance and conditioned feeding behaviour of rats. In the CCK-8-NS and CCK-8-SE treated animals the acquisition of active avoidance and conditioned feeding behaviour were considerably impaired compared to the control; furthermore, these peptides caused a facilitated extinction of active avoidance and conditioned feeding behaviour. The data suggest that cholecystokinin octapeptide is capable of modifying the fear and hunger motivated behaviours of rats. PMID- 6282049 TI - Comparison of computed tomography and angiography in the evaluation of soft tissue tumors of the extremities. AB - CT and angiography were performed in 21 patients with soft tissue tumors of the extremities (18 malignant and 3 benign) and the results were compared with the surgical and histopathologic findings. The tumor size was correctly evaluated at CT in 15 of 21 cases and at angiography in 17 of 21 cases. The main reason for overestimation of longitudinal tumor extent seems to be edema surrounding the poles of high grade malignant neoplasms. CT was superior to angiography in demonstrating the intra- or extramuscular tumor location. If only factors important for the surgical planning are considered, angiography yielded such information in 8 cases while CT was of critical value in the preoperative evaluation of 14 patients. It is concluded that CT should precede angiography in the examination of soft tissue tumors of the extremities. Angiography may be superior to CT in demonstrating vascular anatomy in relation to tumors of the medial thigh. With other tumor locations, angiography should be reserved for those lesions where vascular relationships are not adequately demonstrated at CT. PMID- 6282050 TI - Idiopathic chronic active hepatitis and hepatic cell carcinoma. PMID- 6282051 TI - [The value of FSH, LH, estradiol and testosterone after administration of Seda Presomen in women after hysterectomy with bilateral adnexectomy]. PMID- 6282052 TI - Giovanni Lorenzini Foundation Lecture. Antidepressant drug research: its impact on neurobiology and psychobiology. PMID- 6282053 TI - The effects of subchronic antidepressant drug treatment on the secretion of adenohypophyseal hormones and of corticosterone in the male rat. PMID- 6282054 TI - In vivo effects of typical and atypical antidepressants on brain noradrenergic system: behavioural correlations. PMID- 6282055 TI - Effects of some atypical antidepressants on catecholamine synthesis and release. PMID- 6282056 TI - Stereospecificity of behavioural and biochemical responses to oxaprotiline--a new antidepressant. PMID- 6282057 TI - Adaptive changes as the mode of action of antidepressant treatments. PMID- 6282058 TI - Neurochemical and neuropharmacological investigations into the mechanisms of action of bupropion . HCl--a new atypical antidepressant agent. AB - In the present study, bupropion has been shown to be effective in several behavioral models predictive of antidepressant activity suggesting that it should be an effective antidepressant in man. Furthermore, the data also show that the antidepressant activity of the drug cannot be due to its ability to inhibit MAO present in brain or to increase the release of biogenic amines from nerve endings. It also appears unlikely that the weak properties of the drug as an inhibitor of catecholaminergic pumps in brain csn explain its antidepressant activity. However, the weak but selective block of dopaminergic pumps observed in vivo can be correlated with the mild CNS stimulant properties observed in rodents. Bupropion, failed to desensitize beta-adrenergic receptors in rat cerebral cortex in chronic studies and exhibited equivocal results in acute studies. These neurochemical properties of bupropion serve to distinguish it from typical antidepressants of the MAOI and tricyclic classes and suggest that it should be classified as an atypical antidepressant, whose mechanism of action must still be elucidated. PMID- 6282059 TI - Viloxazine and neurotransmitter function. PMID- 6282060 TI - On the mode of action of mianserin. PMID- 6282061 TI - Effects of zimelidine on various transmitter systems in the brain. PMID- 6282062 TI - Electrophysiological and biochemical comparison of the acute and chronic effects of lithium and tricyclic antidepressants. PMID- 6282064 TI - The importance of the amygdala in the effect of antidepressants on olfactory bulbectomised rats. PMID- 6282063 TI - Modulation of the beta-adrenergic receptor-adenylate cyclase system following acute and repeated treatment with antidepressants. PMID- 6282065 TI - Effect of chronic antidepressants on rat brain alpha 2-adrenoceptor sensitivity. PMID- 6282066 TI - Rapid desensitization of central beta-receptors and up-regulation of alpha 2 receptors following antidepressant treatment. PMID- 6282068 TI - Prolactin and prolactin receptor interactions in normal and neoplastic tissue. PMID- 6282067 TI - Antidepressant drugs: effects on beta-adrenergic and serotonineregic receptors. PMID- 6282069 TI - AXC rat prostatic adenocarcinoma: characterization of cells in culture. AB - We have described the establishment of AXC rat prostatic cancer cells in continuous culture. When injected into isogeneic male rats, these cells produce prostatic adenocarcinomas. The response of androgen and prolactin receptors and ODC in LSC-AXC prostatic cancer cells and tumors to androgen ablation is indistinguishable from that of ventral prostate. In addition, LSC-AXC prostatic tumors retain levels of secretory acid phosphatase comparable to those of ventral prostate of aged AXC rats. These data demonstrate that LSC-AXC prostatic cancer cells and tumors retain a high degree of differentiation, androgen regulated function. The LSC-AXC prostatic cancer cells and tumors appear to represent a unique model system for combined in vivo and in vitro studies of androgen regulation of prostate cancer cell function. PMID- 6282070 TI - The effects of atropine on anaphylactic shock in the guinea-pig. AB - The effects of atropine, 2 mg/kg i.v., on anaphylactic shock were studied in guinea-pigs sensitized to ovalbumin. Atropine only moderately reduced (--31%) the increase in pulmonary resistance observed and slightly prolonged (+26%) the survival time in pretreated animals compared with controls. These effects, however, were no statistically significant. The drug temporarily improved ventilation but had no influence on haematosis. On the other hand, atropine significantly reduced the amount of histamine released (--60%) and of GMPc synthetized in the lung (--21%). The levels of AMPc and prostaglandins E1, E2 and F2 alpha remained comparable to those of control animals. These results suggest that the reflex-induced action of the cholinergic system during anaphylaxis primarily affects large-calibre airways and that the role of acetylcholine in severe reactions is moderate when compared with the direct action of other mediators. PMID- 6282071 TI - Pharmacological interactions between ranitidine, cimetidine, metiamide and tiotidine at histamine H2-receptor sites in guinea-pig atria. AB - Cumulative concentration-response curves to histamine or dimaprit were constructed on guinea-pig isolated right atria and agonist dose-ratios were determined following addition of ranitidine, cimetidine, metiamide or tiotidine alone or a combination of any two of these H2-receptor blocking drugs. The observed histamine or dimaprit dose-ratios for combinations of any two of the H2 antagonists tested were consistent with results predicted from the equation, DR1+2 = DR1 + DR2 - 1, for two antagonists competing for the same receptor sites. Therefore we conclude that all four H2-antagonists compete for the same histamine H2-receptor. PMID- 6282072 TI - Impromidine-induced hypothermia in rats: effect of cimetidine and mianserine. AB - Impromidine, a highly potent histamine H2-receptor agonist, given i.v. at doses of 3.1-62 nmole, induced a dose-dependent hypothermia in the rat with a maximal effect after 15 min. Cimetidine, an H2-receptor antagonist, having no effect when administered alone, antagonized the hypothermic action of impromidine. Two antiserotoninergic agents, p-chlorophenylalanine and metergoline, and chronic treatment with an antidepressant mianserine reduced the impromidine-induced hypothermia. It is suggested that the impromidine-induced hypothermia is an H2 receptor-mediated phenomenon, and the antagonizing effect of mianserine is related to serotonin receptor blocking activity of the drug rather than to its direct H2-receptor antagonism. PMID- 6282073 TI - Effect of purine nucleotides and other compounds on the uptake of histamine and histidine. AB - The uptake of 14C-ring labelled histamine and histidine was studied in human and guinea-pig leucocytes, and in rat peritoneal mast cells. Histamine uptake by sensitized human leucocytes was partly released by antigen or anti-IgE challenge, suggesting that histamine is taken up by the same cells that synthesize and secrete that amine, i.e. basophils. Histamine antagonists, particularly of the H2 subclass, had an inhibitory effect, but histamine agonists had a relatively small and inconsistent effect. Adrenoceptor stimulants and phosphodiesterase inhibitors produced small effects, but dibutyryl cAMP at a concentration of 4-10 mM consistently increased histamine uptake by more than 100% during a 30 min incubation. By contrast, ATP exerted an inhibitory effect, starting at a concentration of 0.2 mM and reaching a maximum (90% inhibition) at 10 mM. Histidine uptake was inhibited by ATP and slightly stimulated by cAMP. Propranolol caused stimulation of histamine uptake and inhibition of histidine uptake at micromolar concentrations. These results suggest that the uptake of histamine is not due to simple diffusion. Although it does not contribute significantly to total cell histamine content or to the removal mechanism of extracellular histamine, it may contribute to the auto-regulatory processes modulating histamine release, synthesis and metabolism. It may also have a significant effect on the extracellular level of histamine, under the influence of drugs or in pathological states. PMID- 6282076 TI - [Wilms' tumor: our experience in 10 years of clinical practice]. PMID- 6282075 TI - [Increased intraocular pressure and the role of phosphodiesterase (author's transl)]. PMID- 6282077 TI - Isolation of Aujeszky's disease virus from boar semen. PMID- 6282074 TI - Comparison of the effects of antioxidant non-steroidal anti-inflammatory drugs against myeloperoxidase and hypochlorous acid luminol-enhanced chemiluminescence. AB - The interaction of myeloperoxidase (MPO) with H2O2 and Cl- provides a potent antimicrobial/cytotoxic system for polymorphonuclear leukocytes (PMNs). MPO related cytotoxicity may be associated with the formation of toxic oxidant MPO intermediates, HOCl, or both. MPO itself is able to oxidize drugs and cellular components. Non-steroidal anti-inflammatory drugs (NSAIDs) able to act as antioxidant free radical scavengers have recently been shown to inhibit luminol enhanced chemiluminescence (CL) which results from the MPO-H2O2-Cl- reaction. CL is a measure of the activity of this reaction. At that time it was not clear whether the source of CL which these NSAIDs affected was HOCl or components of the initial MPO-H2O2-Cl- reaction. A NSAID antioxidant mechanism could affect MPO oxidant intermediates and HOCl. This study compares the effects of antioxidant NSAIDs, methylprednisone and free radical scavengers against MPO-based and NaOCl based luminol-enhanced CL. Most NSAIDs which affected both MPO and NaOCl-CL appeared to share similar mechanisms, suggesting that MPO oxidant intermediates and HOCl are susceptible to NSAID effects. However, most NSAIDs were more effective against MPO-CL. The effect of these NSAIDs against MPO-CL followed the profile of NSAIDs effective in previous studies against PMN-CL. One exception to this was methylprednisone, which has no effect on PMN or MPO-CL, yet inhibited NaOCl-CL. This and other data suggest that MPO and not HOCl-related reactions are a major source of PMN-CL. Less effective NSAIDs affected NaOCl-CL better than MPO CL. While both HOCl and MPO oxidant intermediates may be affected by NSAIDs, it appears that MPO oxidant intermediates or MPO itself are the primary target for NSAID antioxidant free radical scavenging mechanisms. These antioxidant effects impair the major killing system of the PMN and may be NSAIDs' primary anti inflammatory mechanism. Although our data suggests the production of superoxide anion and hydroxyl radical from the MPO-H2O2-Cl- reaction, the actual presence or involvement of these free radical species is not confirmed herein. PMID- 6282078 TI - Diagnostic identification of Aujeszky's disease by an immunodiffusion technique. PMID- 6282079 TI - Effect of adrenocortical function and microclimate on immunoglobulin absorption in calves. PMID- 6282080 TI - Neonatal septicaemia in the tropics. AB - This review has shown a high incidence of neonatal septicaemia in a tropical environment. The predisposing factors leading to increased incidence of primary septicaemia include low maternal socioeconomic status, lack of adequate prenatal care, unsterile delivery practices in those born at home and increased incidence of susceptible hosts. Apart from gram-negative organisms, Staphylococcus aureus was the next common agent responsible for neonatal septicaemia. A combination of penicillinase-resistant penicillin and an eminoglycoside in particular gentamycin, should be the initial antibiotic of choice in suspected cases of neonatal septicaemia. Suggestions for reduction in the incidence of neonatal septicaemia were discussed. PMID- 6282081 TI - The relationship between age and sensitization to PPD-S and atypical mycobacterial antigens among school children in Lagos, Nigeria. AB - A random sample of school children was tested with PPD-S and one of the following antigens simultaneously namely PPD-F, PPD-G, PPD-PL, and PPD-A. The antigens were prepared from the human tubercle bacillus, Mycobacterium Fortuitum, M. Kansasii the Gause Organism, M. Marinum and M. avium respectively. It was observed that for those who had had no BCG, age had a significant effect on the induration size to PPD-S. The age effect was a linear regression effect, the F for regression being significant at the 1% level. The regression coefficient was found to be 0.4 and the equation to the regression line was Y = -0.6 + 0.4X where X is age and Y is induration size. For all the other antigens, age had no significant effect on the size of induration. PMID- 6282082 TI - Differential tuberculin testing in Lagos. AB - School children in lagos were tested with PPD-S and one of the following tuberculins simultaneously namely PPD-G, PPD-A, PPD-Y, PPD-F, and PPD-PL. The tuberculins were prepared from Mycobacterium tuberculosis, the Gause organism, M. avium, M. Kansasii, M. fortuitum and M. marinum respectively. It was found that for the different antigens, among those children who had indurations from 0 to 4 mm those small sized reactions wer generally due to sensitization by opportunist mycobacteria. In contrast to this, among those who had indurations over 10 mm, the reactions were due to sensitization by the human tubercle bacillus. Testing of patients with bacteriologically and radiologically proved tuberculosis showed that practically all of them reacted in an expected manner, i.e. PPD-S elicited larger reactions than PPD-Y, PPD-A, PPD-G, PPD-LL and PPD-F. PMID- 6282084 TI - Congenital aneurysm of the pulmonary artery: a case report. AB - A case of congenital aneurysm of the left pulmonary artery in a 52-year-old Nigerian woman is presented. She presented with a transient history of cough and epigastric pains. A chest radiograph revealed a rounded left hilar mass just superior to the left main bronchus, and pulmonary angiography confirmed the presence of fusiform aneurysm of the left pulmonary artery. At cardiac catheterization, there was pulmonary hypertension (80 mmHg). There was no associated congenital cardiac malformation. The electrocardiograph showed right ventricular hypertrophy. The patient remains asymptomatic to date. This is the fourth known report of pulmonary aneurysm in an African. The rarity of pulmonary artery aneurysms is stressed. PMID- 6282083 TI - Gaucher's disease: a clinical, morphological and biochemical study of a Nigerian family. AB - Two Nigerian sibling presented with progressive hepato-splenomegaly in infancy from which they subsequently died. Morphological investigations carried out showed that the children had Gaucher's disease. Leucocyte B-glucocerebrosidase activities from the parents of the two siblings and also from one surviving sibling were found to be only about 50% of the enzyme activities in control subjects while hexosaminidase and B-glucuronidase activities were within normal limits. There results showed that the three surviving relatives are heterozygotes and also provided a strong supporting evidence that the deceased children had Gaucher's disease. The occurence of this disease in African children living in Kenya, Uganda, the Congo and Nigeria, and in Black Americans suggest that this biochemical abnormality is not as rare in the Negroid race as has been believed hitherto, and may be widespread on the African continent. PMID- 6282085 TI - Studies on the immunopathology of the nephrotic syndrome associated with Plasmodium malariae. 1. Serum levels of an immune adherence inhibitor. AB - A comparative study of the serum levels of an immune-adherence inhibiting factor was carried out on serum samples from eighty-seven nephrotic syndrome children, twenty-eight nephrotic adults, 202 normal school children, 116 blood donors, twenty-five falciparum malaria children and 172 patients with miscellaneous diseases. Low titres (1/32 and below) of the factor were present in sera from 63.2% of the nephrotic children 60.7% of nephrotic adults and 60.0% of children malaria, as compared with 30.7% of the normal children, 25.5% of the patients with miscellaneous diseases and 41.4% of the blood donors. There is a significant difference between nephrotic children and normal children with low titres (P less than 0.05). Furthermore, 36.8% of the nephrotic children had serum titres of 1/4 or less, as compared with 6.4% of normal children. The serum factor is tentatively referred to as 'C3b-inase'. Its similarity to conglutinogen activating factor (KAF) and its possible role in the pathogenesis of the immune complex nephropathy of childhood nephrotic syndrome associated with malaria are discussed. PMID- 6282086 TI - Transferable drug resistance in Pseudomonas patients with premature rupture of membranes in Ile-Ife, Nigeria. AB - Pseudomonas aeruginosa was cultured in eight patients with premature rupture of membranes in a Maternity Unit of the University of Ife Teaching Hospital Complex within a period of 48 h. Three of the strains were found to harbour R-factor DNA conferring high-level resistance to benzylpenicillin, streptomycin, chloramphenicol and tetracycline, while two strains are able to transfer this drug resistance en bloc to drug-sensitive Escherichia coli. The significance of this in the light of a continuing study of drug resistant bacteria in Ile-Ife is discussed. PMID- 6282087 TI - Chemotaxis Methodology: a review. AB - Chemotaxis has gained some prominence in recent years and available today are various methods that are applicable to both in vitro and in vivo measurements of the phenomenon. Some of these methods are discussed. Many are adaptable for use in the clinical laboratory as an aid in the diagnosis of disease syndromes which are characterized by or due to disorders of leucocyte chemotaxis. PMID- 6282088 TI - Cryohaemorrhoidectomy-preliminary experience. AB - Twenty patients had their haemorrhoids treated by cryosurgery and were followed up for periods ranging from 6 to 18 months. The main advantages of this technique were that the periods of hospitalization and convalescence were short, reliance on an anesthetists was less, the major symptom of bleeding was eliminated and the period of postoperative discharge not as long as in other reported series. However, patients with third degree haemorrhoids tended to have a persistent skin tag and depigmentation of the perianal skin was not uncommon. PMID- 6282089 TI - Echocardiographic features of congestive cardiomyopathy in Nigerians. AB - In this study, we compared the echocardiographic features of thirteen patients with chronic congestive cardiomyopathy (COCM) with those of twelve normal subjects. Characteristically, the patients with COCM had larger than normal left ventricular internal dimensions and estimated volumes in enddiastole and endsystole. Qualitatively the septum had a diminished motion, or was flat, similarly the motion of left ventricular posterior wall was less than normal. The diastolic closure rate of the anterior mitral valve (AMV) was less discriminating between normal and COCM patients. However, the distance between the C - point on the AMV and the left side of the septum was significantly greater than normal in COCM. The excursion of AMV was less than normal. Both leaflets of the mitral valves were recorded easily. Although these features are known, they have not been previously reported for Nigerian patients. Echocardiography is useful in the diagnosis of congestive cardiomyopathy, particularly when there are no facilities for Cardiac catheterization. PMID- 6282090 TI - The measurement of blood meal size in Aedes aegypti (L.). AB - Three techniques of estimating blood meal volumes (gravimetric, radioisotope counting and haemoglobin estimation) were compared in adult female Aedes aegypti fed on a cat. Aedes aegypti ingested a mean volume of 2.37 ul, 2.44 ul and 2.57 ul as measured by gravimetric, radioisotope and haemoglobin estimation techniques respectively. This difference was not statistically significant (P greater than 0.05, n = 58). Neither calendar age (post-eclosion) nor strain differences appear to influence blood meal intake but a smaller amount of blood was ingested after a single gonotrophic cycle. The relative merits of the techniques are discussed. PMID- 6282091 TI - Neonatal ascites in Benin, Nigeria. AB - Seven cases of neonatal ascites are presented. The commonest cause is obstructive uropathy from posterior urethral valves which was present in four patients. Two infants had septicaemia and meconium peritonitis was found in the seventh case. Radiological evaluation often revealed the anatomic lesion. The mortality rate was 100%. PMID- 6282092 TI - Is poliomyelitis in the tropics provoked by injections? AB - In the tropics and sub-tropics, poliomyelitis is characterized by a high non seasonal case-rate, very young victims and a predominance of leg paralysis following injections. It is proposes that this could be caused by infection of immunosuppressed infants with poliovirus of low virulence following mal nutrition, infections with malaria and measles and treatment by injections. Vaccination policies are briefly reviewed and the complete safety of inactivated and oral poliovaccines in the tropics is queried on theoretical grounds. Even if injections are coincident with and not causal of paralysis, it may be difficult to persuade parents of this . PMID- 6282093 TI - Dynamic CT of hepatic masses with intravenous and intraarterial contrast material. AB - Seven patients with primary and metastatic hepatic tumors had dynamic computed tomographic scans obtained after an intravenous and an intrahepatic arterial bolus of contrast media. Four patients had hepatoma and three had hepatic metastasis from either a colonic, pancreatic, or leiomyosarcoma primary malignancy. Computed tomography was also performed after an intravenous drip infusion of contrast material. Time-density curves of the hepatic lesions after contrast administration were analyzed and compared. The results demonstrated that: (1) intrahepatic arterial delivery of contrast fluid resulted in the greatest contrast enhancement of lesions and detected more lesions than the intravenous bolus technique, which was superior to the drip infusion technique; (2) no consistent difference in the pattern of contrast enhancement was found between various hepatic lesions; (3) within multiple lesions of similar pathology in any one liver, a spectrum of contrast enhancement pattern was found; and (4) changes in contrast enhancement occurred rapidly and lesions changed from hypodense to isodense to hyperdense to isodense within 30--45 sec. PMID- 6282094 TI - Comparison of pre- and postcontrast CT in hepatic masses. AB - A comparison of hepatic computed tomography (CT) before and after administering intravenous contrast material was performed 85 times on 81 patients suspected of having hepatic mass lesions. Both pre- and postcontrast computed tomography were sensitive and specific in over 88% of cases, with no statistical difference in accuracy between the techniques. However, postcontrast scans were generally preferred because of increased diagnostic confidence and improved characterization of associated abnormalities. The recent advent of rapid sequential scanning after intravenous administration of a large volume of contrast material seems to further improve diagnostic confidence. When high resolution CT equipment is used, postcontrast CT alone is the preferred method for CT evaluation of patients with suspected liver lesions. PMID- 6282095 TI - Sonographic features of placental complications in pregnancy. PMID- 6282096 TI - Nephroblastomatosis: clinicopathologic significance and imaging characteristics. PMID- 6282097 TI - Calcium pyrophosphate dihydrate deposition disease: tumorous form. PMID- 6282098 TI - The ideal radiotracer in gastrointestinal bleeding detection. PMID- 6282099 TI - Atypical Pneumocystis carinii pneumonia in homosexual men with unusual immunodeficiency. AB - Pneumocystis carinii pneumonia is a well known opportunistic infection whose radiographic presentation and association with cytomegalovirus (CMV) have been well described. Recently nine young homosexual men with chronic flulike illnesses were seen. Bronchoscopy in six cases demonstrated evidence of CMW infection with Pneumocystis pneumonia. The radiographic presentation was atypical. Immunologic evaluation revealed T-cell abnormalities. CMV infections altering immunologic mechanisms has been postulated as the underlying cause of this and other uncommon infections in homosexual men. PMID- 6282100 TI - Computed tomography of the liver and spleen with intravenous lipoid contrast material: review of 60 examinations. AB - In 60 computed tomographic examinations of the liver and spleen in 55 patients, a new intravenous lipoid contrast material, Ethiodized Oil Emulsion 13, was used. This organ-specific contrast material, retained by the reticuloendothelial cells of the liver and spleen, caused marked increase in the attenuation of the liver and spleen, but did not appreciably change the attenuation of the tumor involving these organs, thereby significantly increasing the density differential between normal and abnormal tissue. Toxicity was considered to be well within tolerable limits with the most frequently noted untoward side effects of chills, fever, headaches, and foul metallic taste. Ethiodized Oil Emulsion 13 holds the promise to become a valuable clinical tool by which hepatic and splenic imaging may be significantly improved and lesions less than 1 cm in diameter demonstrated in the liver and spleen. PMID- 6282101 TI - Gallium-67 uptake in cystosarcoma phyllodes. PMID- 6282102 TI - Estrogen replacement therapy. PMID- 6282103 TI - Prevention of calcium paradox-related myocardial cell injury with diltiazem, a calcium channel blocking agent. AB - The effect of diltiazem on creatine kinase release and tissue adenosine triphosphate content was investigated during calcium paradox in the isolated perfused rat heart. Creatine kinase loss was minimal during the calcium-free phase, but there was a 100-fold increase in creatine kinase release after reperfusion with normal calcium-containing medium. Diltiazem reduced creatine kinase loss by 35 percent when added to calcium-free medium and by approximately 80 percent when added to both calcium-free and reperfusion media. Adenosine triphosphate content was significantly increased from 2.98 mumol in untreated calcium paradox hearts to 5 mumol/g dry weight in diltiazem-treated hearts. With hypothermia the calcium paradox injury was completely inhibited if the temperature of calcium-free perfusion was maintained at 15 degrees C. Diltiazem appears to exert its protective effect through its ability to prevent the cellular separation and alterations in the gap junctions during calcium deprivation of cells and to limit calcium entry into the cells after reperfusion with calcium-containing medium. PMID- 6282104 TI - Effect of increasing heart rate in patients with aortic regurgitation. Effect of incremental atrial pacing on scintigraphic, hemodynamic and thermodilution measurements. AB - This study was performed to assess the effect of pacing-induced tachycardia in patients with aortic regurgitation. In 12 patients (5 men and 7 women with a mean age of 53 years) with aortic regurgitation, left ventricular end-diastolic and end-systolic volume indexes were measured with multigated equilibrium blood pool imaging, and forward cardiac index was determined with thermodilution, both at rest (mean heart rate +/- standard deviation 72 +/- 8 beats/min) and during atrial pacing at 100 and 120 beats/min. Pacing caused a decremental reduction in left ventricular end-diastolic and end-systolic volume indexes and radionuclide determined stroke volume index but no change in radionuclide-determined cardiac index or left ventricular ejection fraction. Forward cardiac index increased incrementally from the baseline value at rest to that at 120 beats/min despite a decremental reduction in stroke volume index. There was a stepwise decrease in regurgitant volume/stroke (46 +/- 20 ml/m2 at baseline, 27 +/- 15 at 120 beats/min; p less than 0.05) but no change in regurgitant volume/min (3.38 +/- 1.80 liters/min per m2 at baseline, 3.22 +/- 1.78 at 120 beats/min; difference not significant [NS]) or regurgitant fraction (0.54 +/- 0.13 at baseline, 0.49 +/ 0.13 at 120 beats/min; NS). Mean femoral arterial, pulmonary arterial and pulmonary capillary wedge pressures did not change with pacing. PMID- 6282105 TI - Slow release dietary carbohydrate improves second meal tolerance. AB - Breakfasts of lentils or wholemeal bread of identical carbohydrate content were taken by seven healthy volunteers. The lentils produced a significant 71% (p less than 0.001) reduction in the blood glucose area and flattened the plasma insulin and gastric inhibitory polypeptide responses by comparison with the bread. In addition, the lentil breakfast was followed by a significantly flatter blood glucose response to the standard bread lunch which followed 4 h later (by 38%, p less than 0.01). The blood glucose pattern was mimicked by feeding the bread breakfast slowly over the 4 h before lunch. Giving a bread breakfast containing a quarter of the carbohydrate reduced the breakfast glucose profile but resulted in a significantly impaired blood glucose response to lunch (168% of control, p less than 0.01). These results, together with breath hydrogen studies, performed on a separate group of four volunteers, indicate that the flattened response to lentils is not due to carbohydrate malabsorption. Slow release or "lente" carbohydrate foods such as lentils may form a useful part of the diets of those with impaired carbohydrate tolerance. PMID- 6282106 TI - Effect of fiber on breath hydrogen response and symptoms after oral lactose in lactose malabsorbers. AB - The effect of ingesting cellulose, pectin, and psyllium with orally administered lactose in water or milk was tested in six lactose malabsorbers. Breath hydrogen tests were used to evaluate lactose malabsorption and mouth-to-cecum transit times. Addition of psyllium significantly reduced the breath hydrogen response, and symptoms in each subject; whereas, less diminution of expired hydrogen was seen after cellulose or pectin was added. The effect of each fiber on gastric emptying rates of an equal volume liquid meal was evaluated in three volunteers. Pectin had no effect, while the cellulose and psyllium modestly delayed emptying at approximately 30 min. PMID- 6282107 TI - Vindesine in bronchogenic carcinoma: a phase II trial. AB - Twenty-seven patients with advanced bronchogenic carcinoma were treated with vindesine, 3 mg/m2/week. Twenty-three patients were evaluable for response. Two of six patients with small-cell carcinoma and one of 17 patients with non-small cell carcinoma had partial responses. Two other patients with non-small-cell carcinoma had minor responses. The duration of the responses was 2-4 months. Neurologic toxicity occurred in 14 patients and was mild except in two patients. There was a median hemoglobin fall of 2.2 g/dl and a median leukocyte nadir of 2800/microliter during vindesine therapy. Thrombocytopenia occurred in 2 patients and mild thrombocytosis occurred in 10 patients. Seven patients experienced phlebitis or cellulitis at the site of drug administration which could be prevented with small doses of intravenous methylprednisolone. These results suggest that vindesine is well tolerated and possesses some activity in patients with previously treated bronchogenic carcinoma. PMID- 6282108 TI - A phase II trail of vindesine in patients with refractory small-cell carcinoma of the lung. AB - A phase II study of vindesine at a dose of 3 mg/m2 I.V. for 6 weeks and every other week thereafter was carried out in 18 patients with small-cell carcinoma of the lung. All patients were refractory to conventional therapy and all had been treated previously with spindle inhibitors, vincristine, or VP 16-213. All patients were evaluable for response and toxicity. No objective responses were observed. Leukopenia, the only hematologic toxicity, occurred in 13 patients (72%). Neurotoxicity occurred in five patients (28%). Vindesine appears to have limited activity in patients with small-cell carcinoma of the lung previously treated with spindle inhibitors. PMID- 6282109 TI - Hexamethylmelamine as a single agent in the treatment of small-cell carcinoma of the lung. PMID- 6282110 TI - Phase II trial of methyl-CCNU, vincristine, 5-fluorouracil, and streptozotocin (MOF-Strep) in patients with disseminated pancreatic carcinoma. AB - A phase II trial of methyl-CCNU, 5-fluorouracil (5-Fu), vincristine, and streptozotocin (MOF-Strep) was conducted on 20 patients with pancreatic carcinoma. There were two partial remissions (10%) lasting 3 and 10 months. In addition, there were two minor responses. The predominant toxicity was gastrointestinal, although hematologic and renal toxicity were also seen. Since the response rate to MOF-Strep does not appear to exceed that to 5-Fu alone, the usefulness of this combination in patients with pancreatic carcinoma is limited. PMID- 6282111 TI - The modification of melphalan toxicity in tumor bearing mice by s-2-(3 aminopropylamino)- ethylphosphorothioic acid (WR 2721). AB - The toxicity of melphalan in mice was reduced by the injection of S-2-(3 aminopropylamino)-ethylphosphorothioic acid (WR2721). This was seen in terms of reduced toxicity to the stem cells of the bone marrow and intestinal epithelium as well as improved animal survival. Using human melanoma xenografts and growth delay as an end-point, it was demonstrated that WR2721 did not protect this tumor from melphalan. With radio-labelled WR2721, it was shown that WR2721 was rapidly cleared from the blood and actively accumulated by all normal tissues except the CNS. Intact human tumor xenografts and Lewis lung tumors were less able to accumulate WR2721 than normal tissues, but in vitro studies showed that tissue fragments or single cell suspensions of tumors were as efficient as liver fragments or bone marrow cells in accumulating the drug. The rapid clearance of WR2721 and poor vascularity of the intact tumors were thought to be responsible for the differential uptake and protection of normal tissues by WR2721. PMID- 6282112 TI - EBV-IgA and new heterophile antibody tests in diagnosis of infectious mononucleosis. AB - An evaluation has been made of the EBV-IgA tests and the immune adherence hemagglutination assay (IAHA) as compared with standard diagnostic procedures in 119 serial sera from 22 clinical and 113 sera from 42 subclinical cases of EBV infectious mononucleosis. EBV-IgA antibody was demonstrable in 86.4 per cent of patients using EB3 cells as antigen and in 68.2 per cent with P3/HRIK cells. For heterophile antibody the IAHA test was more sensitive, gave higher titers, and was positive longer than the standard absorbed horse or sheep RBC tests in both clinical and subclinical EBV infections. PMID- 6282113 TI - The immunoperoxidase method for rapid diagnosis of Herpes simplex encephalitis (HSE) using touch preparations. AB - Ten rats were inoculated intracerebrally with Herpes simplex virus type I to study the possibility of establishing a rapid diagnosis of Herpes simplex encephalitis through examination of touch preparations of brain tissue stained by the immunoperoxidase method. All except one of the animals died within four days of the developing encephalitis. The surviving animal was killed on the fourth day. Touch preparations were shortly fixed in 4% buffered formalin, cold acetone, or alcohol. The indirect immunoperoxidase method was used to identify viral antigen. The authors found that formalin serves as a good fixative and sensitivity as well as specificity is high. The diagnosis could be made within 4 hours. The cellular morphology is retained. The reaction product is stable and background staining no problem. The results of this study imply to use this method also for rapid diagnosis of brain biopsy tissue in human infections. PMID- 6282114 TI - Concurrence of malignant and benign heterologous mixed tumors of the uterus. AB - A large polypoid uterine mas was composed to two distinct and separate parts: a malignant mixed mullerian tumor (MMMT) and a benign mixed mesenchymal tumor (BMMT). It was considered a collision of two neoplasms rather than a malignant degeneration of the BMMT. Malignant transformation of benign mesenchymal uterine tumors is a controversial concept which is difficult to prove or disprove. Concerning the histogenesis of MMMT it appears, at least in some cases, that they originate not from a single multipotential mullerian cell, but rather from two, a stromal cell and an epithelial cell. It is suggested furthermore, that in certain instances presence of one neoplastic component, e.g., adenocarcinoma, can incite proliferation of the second, sarcomatous element of the MMMT. PMID- 6282115 TI - Foodborne hepatitis A at a family reunion use of IgM-specific hepatitis a serologic testing. AB - A cluster of cases of hepatitis A that occurred among 46 participants in a family reunion in Utah in August 1980 was investigated with epidemiologic and serologic techniques. No index patient with clinical illness had been present at the reunion. Twenty-three cases of hepatitis A were ultimately identified; 14 developed clinical hepatitis. Immunoglobulin M-specific hepatitis A virus antibody testing identified nine recently infected asymptomatic subjects. Eight additional individuals had serologic evidence of prior infection. Detection of these asymptomatically infected and nonsusceptible subjects among presumably well individuals led to the alteration of the food questionnaire analysis, ultimately implicating the green salad. The green-salad maker was identified as an asymptomatically infected individual and was thought to be the probable index case. Determination of hepatitis A virus antibody status of study subjects revealed age-related differences in clinical manifestations and disease susceptibility between individuals exposed to foodborne hepatitis A. PMID- 6282116 TI - Sex partners and herpes simplex virus type 2 in the epidemiology of cancer of the cervix. AB - The authors examined the interaction of exposure to various numbers of sex partners and evidence of antibodies to herpes simplex virus type 2 (HSV-2) in cervical neoplasia in 181 cases of cervical dysplasia, carcinoma in situ and cancer of the cervix and 130 control patients in Los Angeles County, California hospitals, in 1974-1979. Studies by the authors and other investigators have found that risk of cancer of the cervix was enhanced with numbers of sex partners, frequency and duration of using the vaginal douche, early age at first pregnancy, and antibodies to HSV-2 as measured by radioimmunoassay. In this study, it was found that for women with only one or no sex partner in their history, risk was elevated if evidence of antibodies to HSV-2 were present. The same was true for women with two or more sex partners. The risk associated with two or more sex partners was not higher than that for women with one sex partner among those positive for HSV-2, and among those negative for HSV-2. Thus, although this inquiry needs replication on larger numbers of women, whatever the other microorganisms or carcinogens patients were exposed to with multiple sex partners, there was no apparent effect beyond the fact that HSV-2 raises the risk of cancer of the cervix. This may strengthen credence in the hypothesis that HSV 2 is an etiologic factor in cervical cancer. PMID- 6282117 TI - Structure and variation of human ribosomal DNA: the external transcribed spacer and adjacent regions. AB - A group of human ribosomal DNA (rDNA) recombinants that include the probable site for initiation of transcription have been examined for sequence polymorphism. A detailed restriction map of one rDNA insert was constructed using plasmid subclones and end-labeled segments. Comparison of 16 similar rDNA inserts by restriction and heteroduplex analysis demonstrated striking conservation of the external transcribed spacer and 18S gene regions, but defined a region where restriction sites for the enzymes Sma I, Hpa II, and Hha I become frequent or variable. This region extends for about 400--800 base pairs (bp) at the left end of the rDNA insert and is postulated to contain nontranscribed spacer sequences. The use of cloned rDNA segments as probes for the restriction analysis of genomic rDNA has demonstrated certain fixed sites in the nontranscribed spacer that do not vary significantly among different individuals or tumor cell lines. In contrast, restriction with the enzyme Sal I reveals several variable fragments, one of which has been found only in a retinoblastoma cell line. PMID- 6282118 TI - Use of restriction fragment length polymorphisms for genetic counseling: population genetic considerations. AB - Two-locus population genetic models are analyzed to evaluate the utility of restriction fragment length polymorphisms for purposes of genetic counseling. It is shown that the linkage disequilibrium between a neutral marker and a tightly linked overdominant mutant will increase rapidly as the mutant moves to its polymorphic equilibrium. The linkage disequilibrium decays for deleterious recessive mutants. Two measures involving the linkage disequilibrium are investigated to determine how much information the transmission of the neutral marker provides about the transmission of the selected gene. In certain kinds of matings, where the parental two-locus genotypes and linkage phases are known, it is possible to determine whether or not a progeny is homozygous for the selected gene on the basis of the fetal genotype at the marker locus. A quantity of primary interest is the fraction of matings between individuals heterozygous for the selected gene in which exact diagnosis can be made in this way. The expected proportion of such matings, taken over all two-locus matings involving heterozygotes at the selected locus, is calculated as a function of the gene frequencies at the two loci and the linkage disequilibrium between them. This expected value is maximized when the linkage disequilibrium is at its maximum in absolute value. Fewer than half of all matings are informative if the linkage disequilibrium is small in magnitude or if the gene frequencies at the two loci are quite different. Consideration is also given to various conditional measures of association that may be useful when the parental two-locus genotypes are unknown. The results suggest that the utility of tightly linked neutral marker genes in predicting the transmission of a selected gene is generally less when selection acts against a recessive gene than for overdominant selection. PMID- 6282119 TI - Assignment of the human pro alpha 2(I) collagen structural gene (COLIA2) to chromosome 7 by molecular hybridization. AB - A cDNA for the pro alpha 2 chain of human type I collagen has been recently cloned and amplified. We have used this specific probe to identify the human chromosome carrying the pro alpha 2(I) collagen gene. The DNA from 17 independent human/hamster and human/mouse somatic cell hybrids was digested by Eco RI and the restriction pattern analyzed in Southern blot experiments, using the 32P-labeled cDNA as a hybridization probe. The gene coding for the pro alpha 2 collagen subunit could be unambiguously assigned to human chromosome 7. All the other chromosomes, including chromosome 17, were excluded. PMID- 6282120 TI - Spin label electron paramagnetic resonance (EPR) studies of Huntington disease erythrocyte membranes. AB - Several spin-label electron paramagnetic resonance (EPR) studies of red cell membranes appear to show abnormalities in some Huntington disease (HD) patients, but not in others. Both studies measure the W/S ratios, presumably under similar conditions, but have different results. We have studied the W/S ratio in some detail to gain a better understanding of this experimental parameter and to determine its potential application in detecting HD abnormalities. Our results offer little encouragement for continued use of W/S as an indication of membrane defect in HD. PMID- 6282121 TI - Toxicity of metal ions to alveolar macrophages. AB - Significant concentrations of metals are found in the respirable particulate effluents associated with metallurgical smelters. In this investigation the effects of the metallic ions lead, cadmium, iron, mercury, aluminum, chromium, and nickel on various aspects of alveolar macrophage function were studied. The production of antibacterial substances (ie, reactive forms of oxygen) by these cells and oxygen consumption are very sensitive to the metals. Particle uptake displays moderate sensitivity, while lysosomal enzyme activity and membrane integrity are fairly resistant to metals. In addition, the effects of the organic solvents carbon tetrachloride, toluene, and xylene on alveolar macrophage function were tested. These solvents were found to inhibit oxygen consumption and the release of antibacterial substances while not greatly affecting membrane integrity. The results of these experiments indicate that some metals and some organic substances are toxic to alveolar macrophage function. PMID- 6282122 TI - Subclinical lead neuropathy. AB - It had been shown that subclinical impairment of the peripheral nerves may occur in neurologically symptom-free lead workers. In a cross-sectional study, 78 workers from two storage battery plants and one engineering shop were studied; their lead exposure had been monitored with regular blood lead measurements (PbB). The reference group comprised 34 unexposed manual workers. An exposure effect relationship between occupational lead exposure and nerve functions emerged, as nerve conduction velocities decreased with increasing PbBs. The strongest correlations were found between PbB measures and sensory and motor conduction velocities of the median nerve. An exposure-response relationship also emerged as the proportion of subjects with abnormal nerves increased with increasing exposure levels. In a prospective study 24 workers were followed-up from the commencement of their lead work for one year and 16 for two years. The lead exposed showed a tendency of decreasing conduction velocities of arm nerves, but as a group they did not differ statistically significantly from the control group. When the lead exposed were divided into two groups using the median PbBs as the splitting point, the ones with PbBs over 30 micrograms/100 ml showed slowing of serveral nerve conduction velocities, while in the lower exposure the conduction velocities remained at the initial level. Again the clearest difference was noted in conduction velocities of the median nerve. Nerve conduction velocities, especially in the arm nerves slow down in lead exposure and this impairment is caused by really low lead exposure as noted in the prospective follow-up study, which can be regarded as intervention study. PMID- 6282123 TI - Vasopressin secretion induced by hypoxia in sheep: developmental changes and relationship to beta-endorphin release. AB - To investigate the developmental changes in the secretion of vasopressin and the potential role of beta-endorphin as a stimulus to the release of vasopressin, the concentrations of these peptides were measured in fetal, newborn, and adult sheep after episodes of induced hypoxia. The studies confirm that hypoxia is a potent stimulus to the release of both vasopressin and beta-endorphin in the fetal animal. In both the newborn lamb and the ewe, more profound hypoxia is necessary for a similar release. In the fetus, the release of both vasopressin and beta endorphin after hypoxia increased with gestational maturation. A comparison of control concentrations of both peptides, the discordance of release in the newborn lamb, and the absence of a change in concentrations of vasopressin with infusion of beta-endorphin implies that these hormones are released in parallel but independently during hypoxic stress. PMID- 6282124 TI - Prolactin: effects of age, menopausal status, and exogenous hormones. PMID- 6282125 TI - Estrogen-progestin pharmacodynamics of the postmenopausal endometrium studied by thymidine labeling. AB - Five postmenopausal women were treated with conjugated equine estrogens, 1.25 mg tablets for 25 days, and medroxyprogesterone acetate, 10 mg tablets, in combination with the last 10 estrogen doses. Twenty-five endometrial biopsy specimens were incubated in vitro with tritiated thymidine and radioautographic slides were prepared. Within five days of estrogen treatment the thymidine labeling index (TLI) in both glands and stromal cells increased from a very low resting state to relatively high levels of DNA synthesis and cell proliferation. Within five days after addition of progestin, epithelial TLIs decreased to low levels and returned to minimal baseline levels four days after the last steroid dose. Analysis of variances indicated significant changes in epithelial cells (P less than 0.0001) confirming that the proliferative effect of estrogens was suppressed during the progestin phase. Stromal TLI changes were not significant (P = 0.46). PMID- 6282126 TI - Lysosomal enzymes in glomerular cells of the rat. AB - Whole isolated rat glomeruli (WG) were incubated with bacterial collagenase to separate epithelial cells (EC) from the cores of glomerular tufts (GC), which consisted of mesangial and endothelial cells, as demonstrated by electron microscopy. Lysates of WG, EC, and GC and of renal tubules were prepared by hypo osmotic shock and freeze-thawing. Activities of the following acidic lysosomal hydrolases were measured: acid phosphatase, beta-glucuronidase, cathepsin-D, non specific esterase, and aryl sulfatases A and B. The glomerular cell preparations showed activities of all studied enzymes. GC had higher activities than EC, save for nonspecific esterase. Studies of the recovery of acid phosphatase and beta glucuronidase revealed that approximately 2/3 of the hydrolase activities present in WG was still measureable after collagenase treatment and that the bulk of this was found in the GC lysates. These findings demonstrate that the rat glomerulus and its cell components have considerable biochemical activities of acidic hydrolytic enzymes. These appear to be most prominent in the combined mesangial and endothelial cells of the GC components. PMID- 6282127 TI - Human neutrophil peptide receptors: mobilization mediated by phospholipase C. AB - Incubation of human neutrophils with phospholipase C from Clostridium perfringens caused an increase in the ability of the treated cells to bind the chemotactic peptide, F-Met-Leu-Phe. The increase in binding was related to an increase in specific binding of the ligand. The increase in specific binding was, in turn, related to an increased number of peptide receptors. The dissociation constant (KD) for the tripeptide was not altered, on the average, by enzyme treatment. The increase in peptide receptor number was related temporally, and possibly mechanistically, to enzyme-stimulated secretory function involving the secondary granules. Phospholipase C treatment did not similarly augment binding of the complement-derived attractant, C5a. Receptor numbers for different chemotactic ligands may therefore be controlled by different mechanisms. Supplementary experiments provided evidence that this phenomenon was attributable to phospholipase C activity and not to contaminating protease(s). PMID- 6282128 TI - Frequency and distribution of papillomavirus structural antigens in verrucae, multiple papillomas, and condylomata of the oral cavity. AB - Sixty-seven proliferous squamous epithelial lesions of the oral cavity were examined for the presence of human papillomavirus whole (structural) virion antigens by a peroxidase-antiperoxidase technique having immunospecificity against genus-specific (common) antigens of the papillomaviruses. A positive reaction for papillomavirus genus specific antigens was found in 18 of 29 verrucae, 2 of 5 multiple papillomas, and 3 of 5 condylomata; common antigens were not detected in 28 keratoacanthomas. The positive reaction was invariably intranuclear in cells having a focal or diffuse distribution in the superficial epithelium. This study shows that a variety of squamous epithelial lesions of the mucosa are associated with human papillomaviruses and suggests that these viruses may play an important role in the etiology of some cases of squamous hyperplasia of the oral cavity. PMID- 6282130 TI - An ultrastructural study of subacute necrotizing lymphadenitis. AB - Fifteen cases of a unique lymphadenitis called subacute necrotizing lymphadenitis were studied electron-microscopically. The large lymphoreticular cells proliferating at cortical or paracortical areas of the lymph nodes mainly consisted of immunoblasts and histiocytoid cells, which were characterized by numerous intracytoplasmic myelinlike inclusions. Such histiocytoid cells seemed to be derived from the immunoblasts. Tubuloreticular structures, which had been often noticed within endothelial cells or lymphocytes of the patients with systemic lupus erythematosus (SLE) or SLE-related diseases, were also observed with high frequency in most cases examined. They were present within the cytoplasm of immunoblasts, endothelial cells, and histiocytoid cells. Immunoblasts in mitosis occasionally contained these structures. We offer the hypothesis that subacute necrotizing lymphadenitis with still unknown etiology may reflect a self-limited SLE-like autoimmune condition induced by virus infected transformed lymphocytes. PMID- 6282131 TI - Immunopathology of B-cell lymphomas induced in C57BL/6 mice by dualtropic murine leukemia virus (MuLV). AB - Combined clinicopathologic and immunomorphologic evidence is presented that would indicate that a murine leukemia virus (MuLV) with the dualtropic host range is capable of producing a clinically malignant lesion composed of immunoblasts and associated plasma cells in C57BL/6 mice. This process, morphologically diagnosed as an immunoblastic lymphoma of B cells using standard histopathologic criteria, was found to be distinctly polyclonal with regard to immunoglobulin (Ig) isotype when analyzed for both surface and cytoplasmic Ig. Further studies demonstrated that this clinicopathologically malignant, dualtropic MuLV-induced, polyclonal immunoblastic lymphoma of B cells in C57BL/6 mice was normal diploid and unable to be successfully transplanted to nonimmunosuppressed syngeneic recipients. Although all serum heavy and light chain components were found to be progressively elevated as the tumor load increased, the polyclonal increase in serum immunoglobulins was most pronounced for mu heavy and kappa light chains (ie, mu greater than gamma 2A greater than alpha greater than gamma 2B greater than gamma 1; kappa greater than lamba). The dissociation of clinicopathologic and biologic criteria for malignancy in the presently described dualtropic (RadLV) MuLV-induced B-cell lesion is sharply contrasted with the thymotropic (RadLV), MuLV-induced T-cell lymphoblastic lymphoma in C57BL/6 mice. This process is also a clinicopathologically malignant lesion but, when one uses biologic criteria, is found to be distinctly monoclonal, aneuploid, and easily transplanted to nonimmunosuppressed syngeneic recipients. The close clinicopathologic and biologic similarities of the dualtropic MuLV-induced animal model to corresponding human B-cell lymphoproliferative diseases are stressed. PMID- 6282129 TI - Teaching monograph: pathology of skeletal muscle diseases. PMID- 6282133 TI - Chronic hepatitis and hepatocellular carcinoma associated with woodchuck hepatitis virus. PMID- 6282134 TI - Sodium-calcium exchange and sarcolemmal enzymes in ischemic rabbit hearts. AB - We have investigated alterations in sarcolemmal function that occur during myocardial ischemia. Rabbit ventricles were incubated at 37 degrees C for time periods ranging from 5 min to 2 h. The ischemic tissue was homogenized, and activities of the sarcolemmal enzymes Na+-K+-ATPase, K+-p-nitrophenylphosphatase (K+-pNPPase), and adenylate cyclase were measured in the crude homogenate. Na+-K+ ATPase and K+-pNPPase were substantially inhibited after only 10 min of ischemia, and activities for all three enzymes declined progressively up to 1 h of ischemia, when activities were 37-59% of control. Highly purified sarcolemmal membranes prepared from control tissue and myocardium that had been made ischemic for 1 h showed similar purification of sarcolemmal enzymes, passive Ca2+ binding, and passive permeability to Ca2+. However, the velocity of Na+-Ca2+ exchange in ischemic sarcolemmal vesicles was reduced approximately 50% due to a reduction in Vmax. Although the parallel decline in activities of several sarcolemmal functions might suggest a change in membrane structure, phospholipid and cholesterol contents in ischemic sarcolemma were the same as control. PMID- 6282135 TI - Effect of temperature, nutrients, calcium, and cAMP on motility of human spermatozoa. AB - The motility of human spermatozoa and its regulation were examined on cells isolated from other seminal components and purified into fractions of uniform progressive motility. The percent motile cells and estimates of their translational speed were determined by visual inspection, by stroboscopy, and by photon correlation spectroscopy; microcinematography and gradient centrifugation were occasionally used to clarify discrepancies. The motility of isolated spermatozoa could be maintained for periods up to 24 h at 4 or 37 degrees C; the presence of seminal fluid was not required and even provoked a reversible inhibition at 4 degrees C. Albumin facilitated cell movement between microscopic glass plates but had no effect on progressive motility per se, as evidenced by other techniques. During incubations of up to 2 h, progressive cell motility occurred independently of extracellular glucose and calcium but responded to variations in adenosine 3',5'-cyclic monophosphate and calcium. Dibutyryl cAMP increased forward motility, whereas ethyleneglycol-bis(beta-aminoethylether)-N,N' tetraacetic acid reversibly immobilized the spermatozoa in a calcium-dependent manner; phosphodiesterase inhibition resulted in increased vibration of sperm heads without any effect on progressive motility. Longer incubation periods required the presence of extracellular nutrients. These experiments further demonstrate that several motility measuring techniques should be used in parallel to distinguish the various components of cell movement, to exclude aspecific effects, and to supplement the shortcomings of each individual technique. Such procedure could clarify the various discrepancies that have been reported so far and should lead to a better understanding of the regulation of human sperm motility. PMID- 6282136 TI - Influence of presynaptic receptors on neuromuscular transmission in rat. AB - The presence and physiological significance of acetylcholine (ACh) receptors on motor nerve terminals was examined at the rat diaphragm neuromuscular junction. Intracellular recording techniques were used to monitor end-plate potentials (EPP), miniature end-plate potentials (MEPP), and resting potentials of the muscle fibers. Muscle action potentials were blocked by the cut-muscle technique. Quantal release was determined by the ratio EPP/MEPP, after correcting for nonlinear summation. Blockade of acetylcholinesterase with eserine and neostigmine was tested to determine the influence of residual ACh on transmitter release. Partial blockade of ACh receptors with curare was examined to further clarify the role of these presynaptic receptors. The experiments demonstrate that residual ACh inhibits transmitter release and that blockade of ACh receptors enhances transmitter release. It is concluded that presynaptic ACh receptors exist and that they serve an important physiological function. It is suggested that the presynaptic ACh receptors normally serve to limit transmitter release in a negative feedback pathway. PMID- 6282132 TI - Role of oxygen-derived free radicals and metabolites in leukocyte-dependent inflammatory reactions. PMID- 6282138 TI - Nicotinamide restores phosphaturic effect of PTH and calcitonin in phosphate deprivation. AB - Results of previous studies suggest a potentially important role for nicotinamide adenine dinucleotide (NAD) in the cellular regulation of phosphate transport by the renal proximal tubules. The present clearance studies were performed to evaluate whether intraperitoneal administration of nicotinamide, a precursor of NAD and inhibitor of NAD catabolism, would not only increase phosphate excretion but also restore the phosphaturic response to parathyroid hormone (PTH) in rats fed a low phosphate diet. Rats fed a low phosphate diet were resistant to the phosphaturic effect of PTH, calcitonin, and dibutyryl cAMP (DBcAMP) in spite of the fact that all three agents elicited an increase in the urinary excretion of cAMP. Administration of nicotinamide to rats fed a low phosphate diet increased renal cortical NAD levels, increased phosphate excretion, partially restored the phosphaturic effect of PTH and DBcAMP, and completely restored the phosphaturic response to calcitonin. We conclude that nicotinamide restores the phosphaturic effect of PTH and calcitonin in rats fed a low phosphate diet by acting at a cellular step subsequent to cAMP generation to inhibit tubular reabsorption of phosphate. PMID- 6282137 TI - Glucose-induced proton uptake in secretory granules of beta-cells in monolayer culture. AB - The weak base acridine orange (AO) has been shown to be accumulated by the insulin-containing secretory granules of cultured beta-cells in response to high glucose. Various lines of evidence indicate that this accumulation is due to a pH gradient. Thus ionophores such as monensin and nigericin abolish the glucose induced accumulation, and a high concentration of the weak base, benzylamine, results in swelling of the granules. In the absence of glucose, ATP addition to digitonin-permeabilized cells also results in dye uptake. These data also suggest that a primary or secondary active accumulation mechanism for hydrogen ions exists across the granule membrane. PMID- 6282139 TI - Regulation of PTH receptor-adenylate cyclase system of canine kidney: influence of Mn2+ on effects of Ca2+, PTH, and GTP. AB - Metal ions play important roles in the regulation of the activation of adenylate cyclase. Previous studies have suggested that an important site of action of metal ions is at or closely related to the nucleotide regulatory protein. The present studies examine the nature of the regulation of enzyme activity by divalent cations and the influence of Mn2+ on hormone binding and stimulation of adenylate cyclase. Studies were performed in canine renal cortical membranes. Substitution of Mg2+ by Mn2+ was associated with a progressive decline in the ability of GTP or PTH to stimulate adenylate cyclase activity. Mn2+ did not alter specific binding of an iodinated PTH analogue. However, in spite of the loss of guanine nucleotide stimulation of enzyme activity, the effects of guanine nucleotide on PTH binding were not altered in the presence of Mn2+. Substitution of Mg2+ by Mn2+ abolished the inhibitory effect of Ca2+ on basal adenylate cyclase activity. Similarly, the effects of GTP or PTH to enhance the inhibitory effects of Ca2+ on enzyme activity were abolished in the presence of Mn2+. Since Mg2+ and Ca2+ compete for a common allosteric site and Mn2+ abolished the effects of these cations, it would appear that Mn2+ also competes for the binding site of Mg2+ and Ca2+. The present studies demonstrating that Mn2+ does not affect hormone binding or the actions of guanine nucleotides on hormone binding yet totally eliminates the effect of GTP on enzyme activity indicate that the effect of Mn2+ occurs at the level of the interactions of the nucleotide regulatory component with the catalytic unit. In addition, these data suggest that there are two functionally distinct sites of guanine nucleotides with different ionic requirements. PMID- 6282140 TI - Renal cortex ion composition and Na-K-ATPase activity in gentamicin nephrotoxicity. AB - Abnormalities of potassium and magnesium homeostasis have been reported following the use of gentamicin, and potassium depletion enhances gentamicin nephrotoxicity. The present study investigates these relationships in the dog by assessing changes in renal cortex ion composition and renal cortex Na-K-ATPase activity occurring during gentamicin nephrotoxicity. Gentamicin (15 mg/kg i.m. twice daily) was administered for 4 to 7 days to potassium-depleted or potassium supplemented animals. The results show that gentamicin nephrotoxicity was characterized by a significant reduction in renal cortex content of potassium (17%), magnesium (19%), and phosphorus (12%) in all groups of animals given gentamicin. However, only potassium-depleted animals exposed to 7 days of gentamicin experienced a significant rise in plasma creatinine (from 1.3 +/- 0.1 to 4.3 +/- 1.0 mg/dl). Accompanying this increase in plasma creatinine was a significant rise in the renal cortex content of sodium (from 25 +/- 0.5 to 27.9 +/- 1.7 meq/100 g fat-free dry solid wt) and calcium (from 1.2 +/- 0.1 to 2.6 +/- 0.3 mM/100 g fat-free dry solid wt). Na-K-ATPase activity in the renal cortex fell only in potassium-depleted animals after 4 days (from 11.5 +/- 0.9 to 7.8 +/ 0.1 microM Pi.mg protein-1.h-1) and 7 days (5.9 +/- 0.8 microM Pi.mg protein-1.h 1) of gentamicin treatment. Thus, gentamicin nephrotoxicity is characterized by sequential changes in renal cortex ionic composition, sodium pump activity, and renal function. PMID- 6282141 TI - Control mechanisms of bicarbonate transport across the rat proximal convoluted tubule. AB - Bicarbonate transport (JHCO3) was studied in rat proximal convoluted tubules by luminal and peritubular microperfusion, and the effects on tubular bicarbonate transport of selective changes in luminal and peritubular bicarbonate concentrations and of changes in luminal flow rate were evaluated. A pH glass electrode was used to measure [HCO3(-)] and gave results similar to those of a microcalorimetric method. Increasing the tubular and peritubular [HCO3(-)] at constant luminal perfusion rate (10 nl.min-1) augmented JHCO3, but JHCO3 increased more when pH changes were prevented by PCO2 adjustments (constant peritubular pH) than when pH was allowed to rise with the increase in [HCO3(-)] (constant PCO2). Elevation of the tubular HCO3(-) load by raising [HCO3(-)] stimulated JHCO3 more than when the HCO3(-) load was raised by enhancing luminal perfusion rate at constant [HCO3(-)] An increase in PCO2 at constant peritubular pH increased JHCO3. Diamox and benzolamide inhibited JHCO3 at luminal concentrations of 2-4 X 10(-4) M, yet a small but significant fraction of JHCO3 remained intact. Capillary perfusion with 4-acetamido-4'-isothiocyanostilbene 2,2'-disulfonic acid (5 X 10(-4) M) depressed JHCO3 by 70%. Acute changes in luminal and peritubular potassium concentrations (range, 2-6 meq/liter) had no effect on JHCO3, but JHCO3 increased moderately but significantly in severe dietary hypokalemia. PMID- 6282142 TI - Mechanism for the phosphaturia of NH4Cl: dependence on acidemia but not on diet PO4 or PTH. AB - The effects of metabolic acidosis on renal PO4 handling are controversial. Clearance experiments, therefore, were performed in fasted parathyroidectomized rats 1) to test the thesis that NH4Cl per se alters PO4 reabsorption, 2) to characterize the mechanisms responsible for these changes, and 3) to define the interaction of NH4Cl with parathyroid hormone (PTH) in PO4 deprivation. NH4Cl increased the clearance and fractional excretion of PO4 (FEPO4) without altering plasma PO4. Lactic acid and HCl, but not saline loading. NH4HCO3, or glutamine, produced similar effects. At steady-state phosphaturic effects of NH4Cl, neutralization of the acidemia by NaHCO3 abolished the increment. PTH (3.3 U.kg 1.h-1), superimposed on the maximal effective dose of NH4Cl (5.7 mmol.kg-1.h-1), further augmented FEPO4 (from 24.3 to 46.9%). The effects of NH4Cl (delta FEPO4 = 23 vs. 21%) and the synergism with PTH were not affected by PO4 deprivation. In both PO4 repletion and deprivation, NH4Cl increased basal and PTH-stimulated cAMP excretion, but the changes were poorly correlated with FEPO4. We conclude that NH4Cl inhibits PO4 reabsorption independent of PTH, extracellular fluid volume, natriuresis, NH4+ ion, plasma PO4, or the status of PO4 balance. The effects are mediated by mechanisms dependent on acidemia but are quite distinct from those of PTH. Our findings on cAMP are most compatible with the hypothesis that biochemical events beyond cAMP generation mediate both the phosphaturia of NH4Cl and its ability to restore PTH sensitivity in PO4 deprivation. PMID- 6282143 TI - Gastrin stimulation of isolated gastric glands. AB - The ability of gastrin to stimulate acid formation was studied in gastric glands and isolated parietal cells obtained from rabbit gastric mucosa. Accumulation of the weak base aminopyrine and increases in oxygen consumption were used as measures of acid secretory activity. The responses to gastrin were found to be very small (10-15% increase). However, inclusion of dithiothreitol (0.5 mM) in the incubation medium enhanced the responses in both glands and isolated cells to easily detectable levels. For the gastric gland preparation, gastrin stimulation was maximal at 1 X 10(-7) M, with an apparent ED50 of 5 nM. The response reached a maximum at about 30 min and was stable for at least an hour. The gastrin response was enhanced by the phosphodiesterase inhibitor isobutylmethylxanthine and partially inhibited by cimetidine, a histamine H2-receptor antagonist. Combinations of gastrin and histamine showed an additive response over a wide range of histamine concentrations. However, time-course studies revealed a transient potentiation of gastrin response by histamine, which reached a peak at 15 min and was reduced to an additive response by 30 min. Studies using isolated cell populations enriched in parietal cells (approximately 70%) revealed a gastrin stimulation that was not inhibited by cimetidine. The transient potentiation of the gastrin response by histamine was also found in the isolated cell preparation. Gastrin had no effect on cellular cAMP levels or adenylyl cyclase activity. The results are interpreted to indicate that gastrin stimulates acid secretion through three separate actions: 1) a direct stimulation of parietal cell activity, 2) a potentiating interaction with histamine, and 3) for more intact preparations, a release of histamine, which in turn acts as a paracrine stimulus. Quantitatively, the most important action appears to be the release of histamine. None of the actions of gastrin appear to involve a change in cAMP metabolism. PMID- 6282144 TI - Transmural variation in the relationship between myocardial infarct size and risk area. AB - To test the hypothesis that the relationship between infarct area (IA) and area at risk (AR) varies in different layers of the left ventricle (LV), we occluded the circumflex coronary artery for 48 h in 20 conscious dogs. AR was determined by postmortem coronary stereoarteriography, and infarct area by pathological examination. Both AR and IA were divided into four layers: posterior papillary muscle (PPM), subendocardium (Endo), midwall, and subepicardium (Epi) and quantified with planimetry. Hemodynamics and regional myocardial flow with tracer microspheres (7-10 micrometers diam) were measured before and after coronary occlusion. IA was closely correlated with AR for PPM (r = 0.96), Endo (r = 0.97), and Epi (r = 0.92). However, the slope of IA/AR for Endo (1.30 +/- 0.08) was significantly steeper (P less than 0.05) than that for Epi (0.89 +/- 0.11); furthermore, the intercepts at zero infarction for PPM (0.5 +/- 0.1% of LV), Endo (4.2 +/- 0.4%), and Epi (0.1 +/- 0.7%) were significantly different from each other. Regional blood flow measurements indicate that the differences in IA/AR in various layers reflected earlier and greater total collateral flow to the noninfarcted AR in the epicardium. Thus IA/AR for the entire LV is a composite representing separate IA/AR specific to various transmural layers of the LV. In addition, this study demonstrates that the lateral border zone between the IA and the AR is minimal (less than 3-5 mm) in the subendocardium and midwall layers of the left ventricle. PMID- 6282146 TI - Intravascular bronchiolo-alveolar tumor (IVBAT): A low-grade sclerosing epithelioid angiosarcoma of lung. AB - Intravascular bronchiolo-alveolar tumor (IVBAT) is a rare and highly distinctive pulmonary tumor of disputed cellular nature. Both epithelial and endothelial differentiation of this neoplasm have been suggested. We have studied multiple nodules of IVBATs from three patients by light and electron microscopy and by immunohistochemical methods for Factor VIII-related antigen (FVIII RAG). Our light and ultrastructural studies are in essential agreement with the previous suggestion of the endothelial nature of the neoplasm and our demonstration of the presence of FVIII RAG in many of the tumor cells offers new evidence strongly supportive of their endothelial differentiation. We believe that IVBAT and a group of extrapulmonary tumors described as epithelioid hemangioendothelioma and endovascular papillary angioendothelioma are similar biologically indolent neoplasms of epithelioid and dendritic endothelial cells characterized by stromal sclerosis, intravascular spread, a low incidence of metastases and slow clinical evolution. Thus, we regard IVBAT as a low-grade sclerosing angiosarcoma of the lung. PMID- 6282145 TI - Correlation of muscle activity with glycogen metabolism in muscle of Ascaris suum. AB - Isolated muscle segments from the parasitic roundworm Ascaris suum were shown to contract when perfused with acetylcholine (ACh). The muscle responded to ACh concentrations of 1 microM and was maximally contracted at 50 microM ACh. In fed muscle segments perfused with saturating levels of ACh the glycogen synthase Ka values for glucose 6-phosphate increased from 0.5 to 0.95 mM. In starved segments stimulated by ACh, the muscle utilized glycogen at a rate that was 1.41 micrograms.min-1.g tissue-1 greater than the saline-perfused controls. The cyclic AMP (cAMP) levels remained relatively constant at 0.34 +/- 0.08 nmol/g muscle during perfusion with ACh. Contraction in the muscle could be inhibited in a dose dependent manner by gamma-aminobutyric acid (GABA). The presence of GABA in starved muscle prevented the decrease in Ka values and phosphorylase activity ratios brought about by glucose. Perfusion of GABA did not change cAMP levels in the muscle. Starved muscle perfused with GABA utilized glycogen at a rate that was 0.41 microgram.min-1.g-1 greater than saline-perfused controls. The results indicated that muscle contraction could be elicited by ACh, and that the energy for this process was derived from endogenous glycogen stores, which were depleted during contraction. Muscle contraction was also correlated with inactivation of glycogen synthase and activation of phosphorylase. These processes appeared to function via a cAMP-independent mechanisms. PMID- 6282147 TI - Acute hemorrhagic conjunctivitis in Southeast Asian refugees arriving in the United States--isolation of enterovirus 70. AB - During July-September 1980, an epidemic of acute hemorrhagic conjunctivitis (AHC) occurred in several refugee camps and transit centers in Southeast Asia. Of 2,356 refugees examined in Bangkok, 200 (8.5%) had conjunctivitis, including 116 (58%) with hemorrhagic signs. Because increasing numbers of refugees were arriving in the United States with conjunctivitis, a program of surveillance and control was implemented. Enterovirus 70, not previously reported from patients in the Western Hemisphere, was cultured from four arriving refugees. A fourfold rise in titer to enterovirus 70 was found in 10 others, either in the United States or Thailand. After control measures were instituted, the prevalence of the United States or Thailand. After control measures were instituted, the prevalence of conjunctivitis in arriving refugees declined from 49.8 per 1,000 to 3.8 per 1,000. Follow-up of cases after arrival in the United States revealed only one possible secondary case. Extensive epidemics of AHC in the Western Hemisphere are most likely to occur following importation into the humid, coastal areas of Central and South America. PMID- 6282148 TI - Experimental studies of Rio Grande virus in rodent hosts. AB - Rio Grande (RG) virus, a new member of the Phlebotomus fever serogroup, was inoculated into wild wood rats (Neotoma micropus) and laboratory-reared cotton rats (Sigmodon hispidus) to determine if these potential hosts could be experimentally infected. Nine of 14 (64%) wood rats became viremic, with titers of circulating virus ranging from 10(2.3) to 10(5.3) plaque-forming units (PFU)/ml and a geometric mean titer of 10(3.7) PFU/ml. Virus was not detected in urine specimens from inoculated wood rats but was found in a single saliva specimen. RG virus was detected in the blood of 1 of 12 (8%) cotton rats. Neutralizing (N) antibody developed in 8 of 9 inoculated wood rats which survived for 30 days postinoculation and in 11 of 12 cotton rats. N antibody was still detectable in 4 of 7 wood rats which survived for 1 year, and all 7 were resistant to rechallenge with the virus, as were 3 wood rats with naturally acquired antibody. High mortality (36%) occurred in inoculated wood rats; whereas low mortality (8%) occurred in cotton rats. The specific cause of death of the rats was not determined. Modes of transmission of the virus in nature are discussed. PMID- 6282149 TI - Warts and cancer: the oncogenic potential of human papilloma virus. PMID- 6282150 TI - Infections by human papillomavirus (warts). PMID- 6282151 TI - Geographic variation of muscle adenylate kinase in the loach Misgurnus anguillicaudatus. PMID- 6282152 TI - Restriction and modification of DNA by a complex protein. PMID- 6282153 TI - Murine cytomegalovirus pneumonia. Description of a model and investigation of pathogenesis. AB - Pneumonitis was produced in CD-1 mice by intratracheal instillation of murine cytomegalovirus. Lethal pneumonia occurred after instillation of virus derived from partially purified salivary gland homogenates, whereas virus attenuated by serial passage in cell culture caused mild disease and no mortality, even though both virulent and attenuated strains achieved comparable peak titers in lung homogenates. Virulence was characterized by rapid association of virus with pulmonary macrophages and early spread of virus from macrophages to susceptible parenchymal cells. This resulted in enhanced viral growth and cellular destruction during the first 5 days of infection. Delay in the early growth of murine cytomegalovirus in pulmonary parenchyma may diminish the destructive effects of virus on the lung and prevent mortality. PMID- 6282155 TI - Cystosarcoma phylloides of the breast: a new therapeutic proposal. AB - An analysis of forty cases of cystosarcoma phylloides of the breast was undertaken to clarify the relationship between histology, surgical treatment and prognosis of this fibroepithelial tumor. Thirty-eight female patients 12 to 85 years of age, treated for this diagnosis over a 17-year period, were studied retrospectively. Histologically malignant lesions were diagnosed in 17 cases: six were treated by local excision, one by subcutaneous mastectomy with prosthetic implant, four by simple mastectomy, three by modified, two by radical mastectomy, and one by biopsy only. Of the 23 histologically benign tumors, 18 were treated by local excision, three had simple mastectomy, and one had subcutaneous mastectomy with prosthetic implant. Three recurrences, observed (7.5%) among patients with benign tumors, were locally excised. Wide local reexcision has controlled the disease to date after average follow-up of 32 months. In the malignant group, metastases developed in four patients (10%). Three of these patients are dead of disease and one is being treated by chemotherapy. Malignant histology seemed to correlate directly to the presence of pain (46.6%), size of the tumor (average 7 cm in diameter), and older age (average 52 years), but there was no correlation with prognosis. Recent reports on the subject advocate wide local excision for small benign lesions and simple mastectomy for larger or malignant ones. Our data indicates that the histologic appearance does not correlate with the clinical behavior of this neoplasm, and the choice of the procedure does not alter the long-term result, provided that the tumor is completely excised. More radical procedures than wide local excision are not justified for cystosarcoma phylloides which behaves more like a soft tissue sarcoma than a breast gland tumor. PMID- 6282154 TI - Pneumocystis carinii and cytomegalovirus pneumonia in a previously healthy adult. AB - A case of Pneumocystis carinii pneumonia and cytomegalovirus infection occurring in a previously healthy adult male homosexual who was not receiving known immunosuppressive therapy is described. This 35-year-old man had been treated with metronidazole and tetracycline for an intermittent diarrheal illness of 5 months' duration. He was then admitted to the hospital where he was found to have a small infiltrate in the left lower lobe, and this process soon involved both lungs. Initially, Pneumocystis carinii infection and later cytomegalovirus infection were diagnosed by lung biopsy and culture. Peripheral blood lymphopenia was present and he had in vivo and in vitro abnormalities of his cellular immune responses. He was unresponsive to Bactrim as well as pentamidine and pyrimethamine therapy and died 7 wk after hospitalization. PMID- 6282156 TI - Bronchogenic carcinoma masquerading as primary esophageal disease. AB - This report describes our experience with six patients with dysphagia as the sole manifestation of radiographic, inconspicuous primary lung cancer and well-defined esophageal lesion by barium swallow. Esophagograms suggested leiomyoma, benign esophageal stricture, duplication cyst, achalasia, and primary carcinoma of the esophagus. Careful evaluation of the chest radiographs in all patients presenting with dysphagia is emphasized. The majority of esophageal findings are subcarinal and bronchoscopy should be considered essential in the workup of these patients. PMID- 6282157 TI - Autosomal dominant mucoid colon carcinoma: a study of a case and a kindred. PMID- 6282158 TI - Infectious diseases mimicking inflammatory bowel disease. PMID- 6282159 TI - [Thrombocytopenia-absent radius syndrome (author's transl)]. AB - A patient affected with thrombocytopenia and bilateral absence of radius is described. Authors present data suggesting an acquired etiology: intrauterine cytomegalovirus infection and X-ray exposure during the ovulatory period. Patient showed some radiological features not previously described: methacarpian synostosis and absence of sternum ossification centers. Anemia was etiologically related to the iron deficiency secondary to bleeding. Chronic diarrhea, a feature common in this syndrome, was caused by cow's milk protein intolerance, other causes of chronic diarrhea, like pancreatic malfunction or disaccharidal intolerance were discarted. Finally comments on the differential diagnosis of the illness, its' evolution and treatment, pointing out the possibility of an intrauterine diagnosis, crucial for a correct genetic counselling are made. PMID- 6282160 TI - [Wilms tumor. Results of treatment in 14 cases (author's transl)]. AB - The results of 14 cases of Wilms tumor, treated according to the SIOP, nephroblastoma trial and study protocols, are presented. Later, an up to date review of the state of knowledge on the problem is done, mainly in relation to histology, treatment and prognosis. PMID- 6282161 TI - Comparison between plasmids of Salmonella and other enterobacteria isolated from the same patients. AB - The ecology of R plasmids was studied in the intestinal flora of 19 patients with salmonellosis without antibiotic treatment. The plasmids found in the Salmonella strains and the accompanying non-pathogenic Enterobacteriaceae were characterized in each patient. We determined the transferability by conjugation, the fi character and the incompatibility group and did enzyme restriction analysis of these plasmids. The results obtained showed that S. typhimurium is the species of this genus with the highest incidence of R plasmids, and Escherichia coli among the non-pathogenic Enterobacteriaceae. The plasmids found in Salmonella are different from the plasmids found in the other Enterobacteriaceae in fi character (50% fi+ in Salmonella and 5% in the other Enterobacteriaceae) and incompatibility group (33% belong to the FII group in Salmonella plasmids and none on the other Enterobacteriaceae), thereby expressing a different origin. PMID- 6282162 TI - Potential roles of the aminopyrimidine ring in thiamin catalyzed reactions. PMID- 6282163 TI - Thiamin pyrophosphokinase: structure, properties, and role in thiamin metabolism. PMID- 6282164 TI - The enzymatic synthesis of thiamin triphosphate. PMID- 6282165 TI - Characterization of normal intestinal thiamin transport in animals and man. PMID- 6282166 TI - Relaxin secretion and relaxin receptors: the linkages. PMID- 6282167 TI - Unsolved problems of relaxin's physiological role. PMID- 6282168 TI - Preparation of fluoresceinylthiocarbamyl relaxin for the demonstration of relaxin receptors. PMID- 6282169 TI - Octadecylsilica and carboxymethyl cellulose isolation of bovine and porcine relaxin. PMID- 6282170 TI - In vitro induction of relaxin secretion in corpora lutea from nonpregnant rats. PMID- 6282171 TI - The role of platelets in the genesis of ischemia. PMID- 6282173 TI - Collagenase activity in squamous cell carcinoma of the human parotid gland. AB - A collagenase obtained from a single squamous cell carcinoma of human parotid gland was purified to homogeneity by procedures including precipitation with ammonium sulfate, gel filtration, ion-exchange chromatography with DEAE cellulose, and extraction from polyacrylamide gel after electrophoresis. The parotid tumor collagenase had a molecular weight of approximately 68,000 and appeared similar to other mammalian collagenases in many respects such as action on collagen and response to common collagenase inhibitors. Analysis of specimens of various human parotid gland tumors revealed that the levels of collagenase activity were higher in squamous cell carcinoma than in nonsquamous tumors. A high level of collagenase activity from squamous cell carcinoma suggests that collagenase may promote local connective tissue destruction associated with tumor invasion and lymph node metastasis. PMID- 6282174 TI - Evidence for presynaptic parasympathetic receptors on nasal blood vessels. AB - Noradrenergic synapses in the heart and several blood vessels have been shown to possess prejunctional receptors that modulate the release of norepinephrine from the synapse. The present experiment attempted to find evidence for presynaptic receptors for acetylcholine that modulated norepinephrine release. From the evidence obtained, it appears that the acetylcholine released from nasal parasympathetic fibers does not directly affect the smooth muscle of nasal blood vessels. Acetylcholine does, however, appear to inhibit the release of norepinephrine from nasal sympathetic nerve terminals. It appears that any nasal vasodilation produced by nasal parasympathetic fibers is caused by acetylcholine acting on an inhibitory, presynaptic, muscarinic receptor on the sympathetic nerve terminals. Acetylcholine would exert its control over nasal vessels by regulating the degree of sympathetic neurotransmitter release. PMID- 6282172 TI - Calcium antagonists and their potential role in the prevention of sudden coronary death. PMID- 6282175 TI - Breast reconstruction with a cryopreserved nipple. PMID- 6282176 TI - [An experimental study of "slowly resorbed suture material" in the tracheal sutures (author's transl)]. PMID- 6282177 TI - Assignment of phosphoglycerate mutase (PGAMA) to human chromosome 10. Regional mapping of GOT1 and PGAMA to subbands 10q26.1 (or q25.3). AB - Human phosphoglycerate mutase (PGAM, EC 2.7.5.3) is under the control of two structural loci that code for subunits A and B. By means of gene-dosage studies, Bucher et al. (1980) have assigned the loci for GOT1 and PGAMA to chromosome 19 of Mus musculus. Because of the known homologies between human and murine chromosomes, gene dosage studies were carried out in erythrocytes from one patient trisomic for the entire band 10q26 and from another patient monosomic for 10q26.2 and q26.3. Results were compatible with the assignment of PGAMA and GOT1 to 10q26.1 (or 10q25.3). PMID- 6282178 TI - Genetics of paraoxonase. AB - Danish family material comprising 1664 unrelated individuals (parents) and 3169 children, as well as 699 grandparents of the same families, were examined for paraoxonase activity. A micro-autoanalyser method, comprising a primary testing in tris buffer at pH 7.5 and, in the case of primarily intermediate individuals, a secondary testing at pH 10, was applied. This gave a better discrimination than testing only at pH 7.5, because individuals around the low mode of the primary activity distribution had their pH optimum at pH 10, while the optimum of individuals around the high was at pH 8.5. By this combined testing all individuals could be unequivocally classified as 'low' or 'high', and the family material was compatible with the low phenotype representing homozygosity for an autosomal recessive gene with a frequency plow = 0.726. Out of 5532 individuals, 5 showed an almost complete lack of paraoxonase activity. PMID- 6282179 TI - Collecting duct function and sodium balance. PMID- 6282181 TI - Rhabdomyolysis and myoglobinuria. PMID- 6282180 TI - Behavioral pharmacology of the endorphins. PMID- 6282182 TI - Growth factors and regulation of cell growth. AB - A new class of polypeptide hormones known collectively as growth factors has been identified. These polypeptides are able to stimulate DNA synthesis and mitosis of cells cultured in vitro. Growth factors have been isolated from several sources, including platelets, submaxillary glands, pituitary, brain, and medium conditioned by cells grown in vitro. Growth factors appear to behave like classic polypeptide hormones. In the cases of epidermal growth factor and nerve growth factor, specific cell membrane receptors have been studied in detail and partially purified. Platelet-derived growth factor and somatomedin-C interact synergistically and apparently, sequentially to promote cell proliferation in vitro. These studies suggest that cell proliferation is controlled by a synergistic interaction among several growth factors and, perhaps, other hormones. Several in vivo roles for growth factors and a role for tumor-cell derived growth factors in neoplasia have been suggested. PMID- 6282184 TI - Clinical removal of iron. PMID- 6282183 TI - Calcium blocking drugs for angina pectoris. PMID- 6282186 TI - Food and drug interactions. AB - A significant contribution could be made to patient care if nutritional biochemists, basic and clinical toxicologists, and pharmacologists in the various fields were to mount the studies needed to understand the nature of food-drug interactions. If only a small fraction of the 120 billion dollars per year spent for food or the 10 billion dollars expended for drugs were allocated for research in this area, advances might be made for the health of the nation. Changes in man's diet produce marked effects on drug metabolism. We know that changing a customary diet to one high in protein and low in carbohydrate increases the rates of metabolism of antipyrine and theophylline, and shifting to an isocaloric diet of low protein-high carbohydrate slows the rates of metabolism of these drugs. Presumably, high-protein-low carbohydrate diets in man resemble the animal studies with high protein diets that show enhanced hepatic drug metabolism. However, numerous studies emphasize the considerable individual variability to changes in human diets; some people have dramatic changes, whereas others exhibit little or no response. Similar individuality has been found in the response to enzyme induction by smoking. Numerous foods and food ingredients affect drug metabolism in human beings and apparently follow the same patterns as found in experimental animal studies with changes in the levels of cytochrome P-450 dependent monooxygenases in the liver and intestine. These changes presumably exert some protective action against environmental carcinogens, cocarcinogens, or promoters. Dietary modifications are brought about by use of weight-reducing diets, vegetarian diets, hospitalization, or post-operative regimens. These diets are often continued for long periods of time and it is likely they result in changes in the metabolism by the body of subsequently administered drugs or exposure to environmental chemicals. Methods are needed to measure inter individual and inter-group differences in metabolism of foreign compounds in order to accurately assess dietary influences on drug metabolism and vice versa. Epidemiologic studies of rigorously selected human populations, coupled with the newer sensitive chemical analytical methods, will provide the necessary data base for these investigations. PMID- 6282185 TI - Sympathetic control of renin release. AB - Sympathetic outflow influences the renal release of renin through modifications of the tonic activity of the renal nerves and the plasma concentration of catecholamines. These influences may initiate changes in the rate of renin secretion or modulate the response initiated by another of the mechanisms that control renin release. Beta-adrenoceptor mediated stimulation of renin release has been demonstrated in vivo, in the isolated perfused kidney and in preparation in vitro. Likewise an array of evidence has accumulated pointing to the existence of alpha-adrenoceptor mediated inhibition of renin release. However, the cellular location, the physiological significance, and even the existence of these alpha adrenoceptors is still disputed. Receptors sensitive to alterations in the vascular volume have been identified in areas of low and high pressure of the circulation. There is evidence that input from both types of receptors may cancel each other, and that to demonstrate experimentally the effects on renin release of the low pressure cardiopulmonary receptors it is necessary to avoid changes in the input from the high pressure arterial receptors, and vice versa. Again there are dissenting voices that disclaim a tonic inhibitory effect of cardiopulmonary receptor initiated impulses on renin release. The majority of the pharmacological evidence identifies the beta-adrenoceptors in JG cells as of the beta 1-subtype. However, some species may make exception to this generalization. As in other tissues, beta-adrenoceptor mediated influences appear to relate to activation of adenylcyclase in the cell membrane. Considerable interest in the role of calcium in the process of activation of renin release has met with some unexpected, though consistent, experimental findings. PMID- 6282187 TI - Drug interactions at the GABA receptor-ionophore complex. PMID- 6282188 TI - 2,3,7,8-tetrachlorodibenzo-p-dioxin and related halogenated aromatic hydrocarbons: examination of the mechanism of toxicity. AB - In this review, we have examined the biochemical and toxic responses produced by halogenated aromatic hydrocarbons and have tried to develop a model for their mechanism of action. These compounds bind to a cellular receptor and evoke a sustained pleiotropic response. In many tissues this response consists of the expression of a battery of enzymes which are, for the most part, involved in drug metabolism, but in other tissues, those which develop toxicity, an additional set of genes is expressed which effects cellular involution, division, and/or differentiation. The toxicity of these compounds appears to be due to the sustained expression of a normal cellular regulatory system, of which we were previously unaware. In future investigations it is hoped that we will learn the nature and physiologic role of this regulatory system. Only then can we hope to understand the mechanism of toxicity of these compounds. PMID- 6282189 TI - Angiotensin converting enzyme (ACE) inhibitors. AB - Angiotensin converting enzyme inhibitors represent a new class of agents which were designed to retain only that unique property. Because of this inhibition, administration of these agents results in the significant reduction of elevated blood pressure of various etiologies and in the amelioration of symptoms associated with congestive heart failure resistant to digitalis glycosides and diuretics. The mechanism(s) of action is not entirely certain but almost certainly resides in the inhibition of ACE. Certainly, the future will see still further advances made in this therapeutic area as well as with other inhibitors of the renin-angiotensin system. PMID- 6282190 TI - The in vitro activity, human pharmacology, and clinical effectiveness of new beta lactam antibiotics. PMID- 6282191 TI - In vitro activity of MK0787 (N-formimidoyl thienamycin) and other beta-lactam compounds against Bacteroides spp. AB - The susceptibilities of 82 strains of the Bacteroides fragilis group to eight beta-lactam compounds, lincomycin, and metronidazole were determined by using an agar dilution technique. MK0787 (N-formimidoyl thienamycin) was the most active compound, inhibiting all strains at a concentration of 1 microgram/ml. Metronidazole was the only other drug of similar activity. Of the beta-lactam compounds, cefoxitin and MK0787 showed uniform activity against all species, whereas most other compounds were relatively less active against Bacteroides distasonis and Bacteroides thetaiotaomicron than against B. fragilis and Bacteroides vulgatus. Using a well diffusion technique, we determined the relative stability of each beta-lactam compound to sonicated cultures of selected resistant strains. Whereas MK0787 was completely stable to inactivation--and with one exception, cefoxitin was also--ceftriaxone, cefotaxime, cephaloridine, and cefoperazone always showed some inactivation, often quite substantial. Moxalactam and ceftazidime were completely stable to some of the enzyme preparations. PMID- 6282192 TI - In vitro and in vivo studies of three antibiotic combinations against gram negative bacteria and Staphylococcus aureus. AB - The activities of azlocillin, cefotaxime, and amikacin alone and in combination were evaluated in in vitro checkerboard studies, in infected neutropenic mice, and in human volunteers. The combination of cefotaxime plus amikacin was more synergistic in vitro than the others against the Enterobacteriaceae tested, and the combination of azlocillin plus amikacin was more synergistic against Pseudomonas aeruginosa and Staphylococcus aureus. Survival of neutropenic mice infected with Escherichia coli and Klebsiella pneumoniae, respectively, was greater with azlocillin plus amikacin (24 of 40 and 11 of 40) and with cefotaxime plus amikacin (21 of 40 and 17 of 40) than with azlocillin plus cefotaxime (22 of 40 and 3 of 40; P less than 0.05). Median serum bactericidal activity in volunteers receiving these antibiotics alone and in combination was greater than or equal to 1:8 with most agents and with all combinations tested against 10 strains each of E. coli, K. pneumoniae, P. aeruginosa, and S. aureus. These data suggest that clinical trials with combinations of azlocillin or cefotaxime plus amikacin deserve further study in febrile neutropenic patients. PMID- 6282193 TI - Lack of interference of guanosine with ribavirin aerosol treatment of influenza A infection in mice. AB - Guanosine or guanosine 5'-monophosphate did not reduce the protective effect of ribavirin aerosol in influenza virus A/Aichi/68 H3N2 infection of mice. Both compounds partially interfered with the action of a wide range of ribavirin concentrations against influenza virus A/WSN replication in cell culture. PMID- 6282194 TI - Clinical pharmacology of cefotaxime in pediatric patients. AB - Cefotaxime is a new cephalosporin with a spectrum of activity which may make it appropriate for use in pediatric patients. In 33 infants and children, administration of cefotaxime resulted in cure or improvement in 97% of patients, with eradication of 94% of isolated pathogens. Toxicity was minimal. The disposition of cefotaxime in this age group was similar to that reported for adults, with an elimination half-life of approximately 1.5 h, a volume of distribution of 1 liter/kg, a total serum clearance of 10 ml/min per kg, and a renal clearance of 6 ml/min per kg. PMID- 6282195 TI - Acyclovir therapy of neonatal herpes simplex virus type 2 infections in rabbits. AB - New Zealand white rabbits less than 30 h old were inoculated subcutaneously with 10(3) 50% tissue culture infectious doses of type 2 herpes simplex virus. The animals were randomly assigned to a treatment schedule of daily intraperitoneal injections of acyclovir, beginning on the day of virus inoculation for 6 or 12 days, on post-inoculation day 1 for 6 days, or on post-inoculation day 2 for 6 days. The acyclovir was given in doses of 50 mg/kg of body weight per day. Similarly infected animals receiving daily intraperitoneal injection of Eagle minimum essential medium served as controls. All of the control animals died on day 4 or 5 after inoculation. At death they exhibited severe skin lesions, viremia, and dissemination of virus in various visceral organs and spinal as well as trigeminal ganglia. In contrast, animals treated with acyclovir failed to develop significant skin lesions, and death did not occur while treatment continued. Termination of treatment after 6 days resulted in late-onset fatal disease and virus isolation from the brain in many rabbits regardless of the treatment schedule. No such late fatality was observed and no virus could be detected from the brain when treatment was initiated on the day of virus inoculation and continued for 12 consecutive days. With respect to all of the variables studied, treatment for 12 days beginning on the day of virus inoculation was most effective. PMID- 6282196 TI - Metabolism of acyclovir in virus-infected and uninfected cells. AB - The metabolism of acyclovir to its mono-, di-, and triphosphate derivatives was examined in uninfected and virus-infected cells. The level of phosphorylation of acyclovir was dependent upon virus type, cell line, exogenous drug concentration, and exposure time. Acyclovir phosphorylation was inhibited by exogenously added nucleosides. The order of inhibition was deoxythymidine greater than deoxycytidine greater than guanosine greater than or equal to deoxyguanosine. Acyclovir triphosphate persisted in infected cells after removal of the drug from the medium. The initial half-life of the triphosphate was 1.2 h in the absence of the drug in the medium, but triphosphate levels reached a plateau after 6 h. The presence of low concentrations of the drug in the medium resulted in a longer persistence of the intracellular triphosphate and a higher plateau level. PMID- 6282197 TI - Inhibition of bluetongue and Colorado tick fever orbiviruses by selected antiviral substances. AB - The effects of four ribonucleic acid virus inhibitors were evaluated in cell cultures and in mice to determine inhibitory effects against bluetongue virus and Colorado tick fever virus (CTFV). Test compounds included 1-beta-D-ribofuranosyl 1,2,4-triazole-3-carboxamide (ribavirin), 3-deazaguanine, 3-deazauridine, and 9 (S)-(2,3-dihydroxypropyl)adenine. Ribavirin-2',3',5'-triacetate (ribavirin triacetate) was evaluated in vivo against CTFV. Inhibition of cytopathic effect and plaque reduction were used to evaluate antiviral activity. In cytopathic effect inhibition studies, bluetongue virus was markedly inhibited by 3 deazaguanine and 3-deazauridine in Vero cells with moderate inhibition by the other agents. Ribavirin and 3-deazaguanine markedly inhibited CTFV in MA-104 cells, 3-deazauridine was slightly less active, and 9-(S)-(2,3 dihydroxypropyl)adenine was negative. Ribavirin was less effective in Vero cells against CTFV. When mice were inoculated intracerebrally with CTFV and treated by a single intracerebral injection with drug, ribavirin triacetate increased the number of survivors, 3-deazaguanine increased mean survival time, and ribavirin was negative. Intraperitoneal treatment of infected mice with ribavirin triacetate for 1 week significantly increased the number of survivors and mean survival time, providing strong evidence that the agent is active across the blood-brain barrier. PMID- 6282198 TI - Serum bactericidal activity of moxalactam and cefotaxime with and without tobramycin against Pseudomonas aeruginosa and Staphylococcus aureus. AB - Serum bactericidal activity was determined against 10 strains each of Pseudomonas aeruginosa and Staphylococcus aureus in serum from volunteers 1 and 6 h after intravenous infusion of cefotaxime, tobramycin, and the combination; or of moxalactam, tobramycin, and the combination. High serum bactericidal activity against P. aeruginosa was found significantly more frequently with moxalactam plus tobramycin than with cefotaxime, moxalactam, and tobramycin alone or with cefotaxime plus tobramycin. PMID- 6282200 TI - Susceptibility of gram-positive cocci to various antibiotics, including cefotaxime, moxalactam, and N-formimidoyl thienamycin. AB - The activities of cefotaxime, moxalactam, MK 0787 (N-formimidoyl thienamycin), ampicillin, oxacillin, vancomycin, and clindamycin were compared against gram positive cocci. MK 0787 was the most active and moxalactam was the least active of these drugs, except against methicillin-resistant Staphylococcus aureus, where vancomycin was most active, and penicillin-resistant pneumococci, where cefotaxime was more active. PMID- 6282199 TI - Activities of new beta-lactam antibiotics against isolates of Pseudomonas aeruginosa from patients with cystic fibrosis. AB - The in vitro activities of gentamicin, tobramycin, amikacin, azlocillin, carbenicillin, mezlocillin, piperacillin, ticarcillin, cefotaxime, ceftizoxime, cefoperazone, cefsulodin, moxalactam, ceftazidime, ceftriaxone, and N-formimidoyl thienamycin were measured against 62 isolates of Pseudomonas aeruginosa obtained from patients with cystic fibrosis. Ceftazidime and N-formimidoyl thienamycin were the most active of these agents. PMID- 6282201 TI - Derivatives of 4-dihydro-4-deoxy-4(R)-amino spectinomycin and their activity against susceptible and resistant Escherichia coli strains. AB - A number of alkyl and acyl derivatives of 4-dihydro-4-deoxy-4(R)-amino spectinomycin were tested against various Escherichia coli strains, possessing different susceptibilities to spectinomycin. The influence of the lipophilicity and the length of the side chain substituents of the derivatives was compared to both minimal inhibitory concentration values and stability to adenyltransferase. Derivatives with a chain length of more than 10 carbon atoms demonstrated a significantly higher activity against all investigated strains, whether susceptible or resistant. The same inhibitory effect was achieved with short chain aminoacyl derivatives only against susceptible strains. Other short-chain derivatives possessed no advantage to spectinomycin. A 10-fold decrease in the affinity for adenyltransferase was achieved in compounds with a high lipophilicity (log P), i.e., in aliphatic substituted derivatives with a log P greater than 4 and in benzoyl-substituted derivatives with a log P greater than 2. Derivatives with branched alkyl chains and long side chains displayed a different mode of action than spectinomycin. They possessed strong activity against strains with an altered ribosomal binding site and a decreased influence of pH on antimicrobial activity. PMID- 6282202 TI - Clinical efficacy of cefotaxime in serious infections. AB - Thirty-five patients underwent 38 treatment courses with cefotaxime. Documented infections included 11 bacteremias, 7 cases of nosocomial pneumonia, 6 surgical wound infections, 3 bone infections, 1 biliary infection, and 1 urinary tract infection. Granulocytopenic patients with fever received 15 courses of empiric cefotaxime therapy alone; in 8 courses, no definite site of infection or pathogen was isolated. Broad-spectrum antibiotics had been administered to 23 patients before cefotaxime. Thirty-seven bacterial pathogens were isolated from 25 patients. Three such pathogens were resistant to cefotaxime and required alternative therapies. Pathogenic isolates included 13 Serratia marcescens, 12 Pseudomonas aeruginosa, 4 Escherichia coli, 2 Klebsiella pneumoniae, 2 Providencia stuartii, 1 Enterobacter cloacae, 1 Haemophilus influenzae, 1 Enterococcus, and 1 Staphylococcus aureus. Of the treatment courses, 25 of 38 resulted in a favorable response to cefotaxime, including 9 of 15 in granulocytopenic patients. Superinfection was seen in one patient. The emergence of resistance was documented in another patient. Of 15 patients with multiply resistant pathogens, 12 improved with cefotaxime. Of 12 patients with Pseudomonas aeruginosa, 6 favorably responded. Possible complications of cefotaxime were observed in 14 of 42 treatment courses. Cefotaxime is most useful in treatment of infections due to multiply resistant, gram-negative pathogens other than Pseudomonas aeruginosa. PMID- 6282203 TI - Intramuscular and intravenous pharmacokinetics of cefmenoxime, a new broad spectrum cephalosporin, in healthy subjects. AB - This study was concerned with the single-dose, pharmacokinetics of cefmenoxime after intramuscular (i.m.) injections of 250, 500 and 1,000 mg; 1-h intravenous (i.v.) infusions of 500, 1,000, and 2,000 mg; and 5-min i.v. injections of 500, 1,000, and 2,000 mg of cefmenoxime. A total of 15 subjects were used, each receiving all three doses for one route of administration. Mean calculated peak plasma levels after the 250-, 500-, and 1,000-mg i.m. doses were 9.07, 14.68, and 26.73 micrograms/ml, respectively, occurring about 40 min after dosing. The biphasic decline in plasma levels after i.v administration was usually not apparent after i.m. dosing, because absorption of the drug from the injection depot was slower than distribution of the drug. Mean calculated peak levels from the 500-, 1,000-, and 2,000-mg i.v. doses were 22.8, 41.6, and 94.5 micrograms/ml, respectively, after the 1-h infusions and 64.1, 100.9, and 198.2 micrograms/ml, respectively after the 5-min injections. Small but statistically significant trends of decreasing alpha and increasing volume of distribution (central compartment) with increasing dose size were noted; however, this distribution phenomenon was self-compensating, resulting in no overall effect on plasma clearance. For practical purposes, the pharmacokinetics were linear. The mean 0- to 24-h urinary recoveries of cefmenoxime after the i.m. injections, i.v. infusions, and i.v. injections were 72.1, 67.5, and 74.5% respectively. Overall, the pharmacokinetics of cefmenoxime were best described by a two-compartment open model with a beta-phase half life of 0.91 h. Plasma clearance of the drug was dosage level and route independent, averaging 254 ml/min; thus, there was an excellent linear relationship between the area under the plasma level curve and the dose. The results of this study indicated that most of the drug is removed by renal mechanisms, with tubular secretion predominating. PMID- 6282204 TI - Disposition of ketoconazole, an oral antifungal, in humans. AB - The pharmacology of ketoconazole was studied in patients with fungal infections. After administration of 50-, 100-, and 200-mg doses of ketoconazole, there was a linear increase in the area under the curve of serum concentrations; this was not apparent when higher doses of ketoconazole were given. An increase in the area under the curve occurred in patients receiving 200 mg daily who were restudied after 1 to 12 months of therapy. However, normalized area under the curve appeared to decrease after higher doses were administered chronically. The half life ranged from 2.0 to 3.3 h. Peak serum concentrations up to 50 micrograms/ml were detected in this study, and potentially therapeutic concentrations were detectable up to 26 h after high doses. Ketoconazole penetrated the saliva and inflamed joint fluid and meninges, although variably, and could be demonstrated in some other tissue compartments. In the presence of renal failure, ketoconazole disposition was not altered, whereas in the presence of hepatic insufficiency, an alteration in disposition was suggested. The interactions of ketoconazole and other drugs were studied. Of note, antacids did not significantly affect ketoconazole pharmacokinetics (nor did meals), and ketoconazole and warfarin did not appear to affect the pharmacokinetics of the other. PMID- 6282205 TI - Plasmid-mediated sulfonamide resistance in Haemophilus ducreyi. AB - Clinical isolates of Haemophilus ducreyi from patients with chancroid were shown to have one or more 4.9- to 7.0-megadalton non-self-transferable plasmids and to have in vitro resistance to sulfonamides. Transformation of Escherichia coli to sulfonamide resistance was associated with the acquisition of a 4.9-megadalton plasmid, which did not confer linked resistance to streptomycin. The guanine-plus cytosine content of this plasmid was found to be 57%. Filter-blot hybridization and restriction endonuclease digestion studies suggested a relationship of this plasmid to RSF1010. Electron microscope heteroduplex analysis confirmed this relationship. The identification in H. ducreyi of a plasmid closely related to plasmids found in enteric species, rather than transposition of a resistance determinant to an indigenous plasmid, suggests that further dissemination of the enteric plasmid pool to this genus is possible since plasmid transfer between certain Haemophilus species is readily demonstrated. PMID- 6282206 TI - In vitro activity of moxalactam and mecillinam, singly and in combination, against multi-drug-resistant Enterobacteriaceae and Pseudomonas species. AB - The in vitro interaction of moxalactam and mecillinam against multi-drug resistant gram-negative enteric bacilli was studied by checkerboard microdilution susceptibility tests and by killing curve kinetics. Against Enterobacteriaceae, the combination was unpredictable; the frequencies of synergy, indifference, and antagonism were 11, 76, and 13%, respectively. Against Pseudomonas sp., the two drugs were consistently indifferent. Overall, the combination of moxalactam and mecillinam was no more active than moxalactam alone. PMID- 6282207 TI - In vitro susceptibility of varicella-zoster virus to E-5-(2-bromovinyl)-2' deoxyuridine and related compounds. AB - The in vitro susceptibility of eight strains of varicella-zoster virus (VZV) to E 5-(2-bromovinyl)-2'-deoxyuridine (BVDU) was examined in human embryonic fibroblasts by the following techniques: inhibition of focus formation by either cell-free VZV (4-day assay) or cell-associated VZV (2-day assay), inhibition of viral antigen formation (2-day assay), and inhibition of viral cytopathogenicity (15-day assay). The 50% inhibitory dose (ID50) of BVDU ranged from 0.001 microgram/ml (2-day assay) to 0.01 microgram/ml (15-day assay). BVDU appeared highly selective in its anti-VZV activity since even at concentrations as high as 100 micrograms/ml, BVDU did not markedly affect the viability of the host cells. The ID50 of BVDU for VZV was comparable to that of IVDU (E-5-(2-iodovinyl)-2' deoxyuridine). Both drugs inhibited the replication of VZV at a much lower concentration than did other antiviral compounds such as iododeoxyuridine, ethyldeoxyuridine, arabinosylcytosine, arabinosyladenine, phosphonoacetic acid, iododeoxycytidine, and acycloguanosine. BVDU and IVDU were virtually inactive against a thymidine kinase-deficient VZV mutant, suggesting that phosphorylation by the viral enzyme is responsible, at least in part, for the selective anti-VZV activity of the compounds. PMID- 6282208 TI - Multiple-dose pharmacokinetics of amdinocillin in healthy volunteers. AB - The pharmacokinetics of amdinocillin (mecillinam) after multiple intravenous doses to healthy subjects are described. Assay of plasma and urine samples was carried out with a sensitive and specific high-pressure liquid chromatographic technique. A dose of 10 mg/kg of body weight was administered every 4 h for six doses. No accumulation was noted. Mean peak plasma concentrations were approximately 50 micrograms/ml, and the plasma half-life was approximately 53 min. Total plasma clearance was 4.6 ml/min per kg after the first dose, which declined slightly to 4.1 ml/min per kg after the last dose. Renal clearance remained fairly constant at approximately 2.9 ml/min per kg, or twice the creatinine clearance. The fraction excreted unchanged totaled 63% during the 4-h interval after the first dose and was nearly 70% overall. The steady-state volume of distribution was calculated to be 0.26 liter/kg. Urinary concentrations of amdinocillin were far in excess of the usual inhibitory concentrations for susceptible pathogens and were as high as 3,000 micrograms/ml. Dose of amdinocillin every 4 h provide plasma and urine concentrations which should be effective for the treatment of infections. PMID- 6282209 TI - Inhibition of rotaviruses by selected antiviral substances: mechanisms of viral inhibition and in vivo activity. AB - Several RNA virus inhibitors were evaluated against simian (SA11) rotavirus infections in vitro and murine rotavirus gastroenteritis in vivo. Test compounds included 1-beta-D-ribofuranosyl-1,2,4-triazole-3-carboxamide (ribavirin), 3 deazaguanine (3-DG), 3-deazauridine, and 9-(S)-(2,3-dihydroxypropyl)adenine [(S) DHPA]. All drugs inhibited total infectious SA11 virus yields in MA-104 cells. Ribavirin, 3-DG, and (S)-DHPA affected [3H]uridine uptake into uninfected MA-104 cells in both the acid-soluble and -insoluble fractions. All drugs reduced the levels of dense (precursor) and light (complete) SA11 particle yields compared with control but did not alter the relative amounts of dense compared with light particles, suggesting that the agents did not interfere with virus assembly. Ribavirin and 3-DG inhibited SA11 polypeptide synthesis, as determined by polyacrylamide gel electrophoresis studies. None of the agents or mono- and triphosphate derivatives of ribavirin inhibited SA11 RNA polymerase activity. In murine rotavirus studies, oral therapy with ribavirin-2',3',5'-triacetate and (S) DHPA increased mean survival time, but no increase in survivor rate was observed. 3-DG- and (S)-DHPA-treated mice had a more rapid weight gain than controls, suggesting a probable lessening of the severity of the disease. PMID- 6282211 TI - Comparative pharmacokinetics and tissue penetration of sulbactam and ampicillin after concurrent intravenous administration. AB - A combination of 500 mg each of sulbactam and ampicillin was administered intravenously into six healthy male volunteers, and subsequent serum and blister fluid levels were measured. Both dogs had similar serum half-lives of about 1 h. The concentrations of sulbactam in serum and blister fluid were about 1.5 times those of ampicillin, and appeared to be pharmacokinetically well matched when administered in equal doses. PMID- 6282210 TI - Regulation of cephamycin C synthesis, aspartokinase, dihydrodipicolinic acid synthetase, and homoserine dehydrogenase by aspartic acid family amino acids in Streptomyces clavuligerus. AB - The effect of the cephalosporin precursors and amino acids of the aspartic acid family on antibiotic production by Streptomyces clavuligerus was investigated DL meso-Diaminopimelate and L-lysine each stimulated specific antibiotic production by 75%. A fourfold increase in specific production was obtained by simultaneous addition of the two compounds. The stimulation could be further increased by adding valine to the two effectors. In the streptomycetes the alpha-aminoadipyl side chain of the cephalosporin antibiotics is derived from lysine. Streptomycetes, like other bacteria, are expected to produce lysine from aspartic acid; therefore, the feedback control mechanisms operating in the aspartic acid family pathway of S. clavuligerus, which may affect the flow of carbon to alpha aminoadipic acid, were investigated. Threonine inhibited antibiotic production by 41% when added to minimal medium at a concentration of 10 mM. Simultaneous addition of 10 mM lysine completely reversed this inhibition. The aspartokinase of S. clavuligerus was found to be subject to concerted feedback inhibition by threonine and lysine. Threonine may act to limit the supply of lysine available for cephamycin C biosynthesis via this concerted mechanism. Single or simultaneous addition of any other amino acid of the aspartate family in the in vitro assay did not inhibit aspartokinase activity. Activity was stimulated by lysine. Aspartokinase biosynthesis was partially repressed by methionine or isoleucine at concentrations higher than 10 mM. Methionine, but not isoleucine, inhibited cephamycin C synthesis by 27% when added to minimal medium at a concentration of 10 mM. Dihydrodipicolinate synthetase, the first specific enzyme of the lysine branch, was not inhibited by lysine but was partially inhibited by high concentrations of 2,6-diaminopimelate and alpha-aminoadipate; it was slightly repressed by diaminopimelic acid. Homoserine dehydrogenase activity was inhibited by threonine and partially repressed by isoleucine. It appears that S. clavuligerus aspartokinase is a key step in the control of carbon flow toward alpha-aminoadipic acid. PMID- 6282212 TI - Characterization of ampicillin resistance plasmids from Haemophilus ducreyi. AB - Seven strains of Haemophilus ducreyi from diverse geographic origins were analyzed for their plasmid content. All strains were multiply resistant, but only resistance to ampicillin was transferred to Escherichia coli by transformation. The H. ducreyi plasmids encoding for ampicillin resistance were 7.4, 5.7, and 3.6 megadaltons and encoded for part or all of TnA, and ampicillin transposon. The relatedness of these plasmids was examined by restriction endonuclease digestion and DNA-DNA homology with isolated DNA fragments from TnA. PMID- 6282214 TI - Comparative efficacy of antiherpes drugs in different cell lines. AB - A selection of antiherpes compounds including bromovinyldeoxyuridine (BVDU), acyclovir (ACV), and fluoroiodoaracytosine (FIAC) were compared for their inhibitory effects on herpes simplex virus type 1 (strain KOS) replication in a wide variety of human, simian, feline, and murine cell lines. The 50% inhibitory doses of the compounds varied considerably from one cell line to another, i.e., from 0.01 to 2 micrograms/ml for ACV and from 0.004 to 0.2 micrograms/ml for BVDU and FIAC. However, the relative order of antiviral potency remained constant, thus (in order of decreasing potency) BVDU greater than FIAC greater than ACV, for all cell lines studied except for the murine embryo cells, in which the order of decreasing potency was BVDU greater than ACV greater than FIAC, and for the feline lung cells, in which the order of decreasing potency was FIAC greater than ACV greater than BVDU. PMID- 6282213 TI - Anion-exchange extraction of cephapirin, cefotaxime, and cefoxitin from serum for liquid chromatography. AB - An anion-exchange column technique for extraction of antibiotics from serum proteins has been developed for use in the assay of cephapirin, cefotaxime, and cefoxitin by high-pressure liquid chromatography. Anion-exchange extraction of cephapirin from serum samples by this technique was compared with protein precipitation methods, using 6% trichloroacetic acid or absolute ethanol. Column extraction gave improved quantitative drug recovery and reduced background serum interferences in the resultant chromatograms when evaluated against protein precipitation. Comparisons of this method with microbiological assay gave statistically equivalent results. Twelve patient samples were assayed for cephapirin, and no interferences were encountered from the 22 systemic agents these subjects were receiving. The anion-exchange technique for antibiotic extraction provides a rapid, precise, and quantitative antibiotic assay when used with liquid chromatography. PMID- 6282215 TI - Antimicrobial susceptibility of Bordetella pertussis strains isolated from 1960 to 1981. AB - The susceptibilities to erythromycin, rifampin, polymyxin B, ampicillin, tetracycline, gentamicin, fusidic acid, trimethoprim, and spectinomycin of 100 strains of Bordetella pertussis isolated between 1960 and 1981 were compared. No change in susceptibility to any of these drugs was noted. PMID- 6282216 TI - Pharmacokinetics of cefotaxime in newborn infants. AB - The pharmacokinetics of cefotaxime were determined in newborn infants who were 1 to 7 days of age. Mean peak serum concentrations of 116 and 132 micrograms/ml were observed at completion of a 10-min intravenous infusion of 50 mg of cefotaxime per kg in low and average birth weight infants, respectively. The mean elimination half-lives were 4.6 and 3.4 h and rates of clearance from serum were 23 and 44 ml/min per 1.73 m2, respectively. A dosage schedule for cefotaxime in newborn infants is presented. PMID- 6282217 TI - Induction of cytopathogenicity in mammalian cell lines challenged with culturable enteric viruses and its enhancement by 5-iododeoxyuridine. AB - Cultures of 17 established cell lines were tested against 105 enteric virus types for capacity to support viral replication as indicated by cytopathogenic effect production. Enhancement of susceptibility by treatment of the cells with 5 iododeoxyuridine was evaluated in parallel with untreated cells. Cytopathogenic effect was produced in two or more cell lines by every virus tested except six strains of group A coxsackie virus. No cell line was found to be susceptible to these six virus types. In general, treatment with 5-iododeoxyuridine provided a more rapid onset of cytopathogenic effect in susceptible cells and in some instances resulted in refractory cells becoming permissive to viral replication. The use of 5-iododeoxyuridine allowed two human embryonic lines (HEL-299 and L 132), in combination, to be susceptible to all but the six group A coxsackie virus strains. PMID- 6282218 TI - Isolation of enteroviruses from oxidation pond waters. AB - Enteroviruses were isolated from influent and effluent wastewaters of two types of oxidation lagoons during an 18-month study. Isolations, performed by sewage concentration and direct assay, were low in number and did not follow a seasonal trend. The younger treatment system, using aeration and effluent chlorination, was more efficient at removing viruses than the aging, static complex. PMID- 6282219 TI - A reaction between the superoxide free radical and lipid hydroperoxide in sodium linoleate micelles. PMID- 6282220 TI - Some properties of purified 3-hydroxy-3-methylglutaryl coenzyme A reductase phosphatases from rat liver. PMID- 6282221 TI - Intact form of myeloperoxidase from normal human neutrophils. PMID- 6282222 TI - Abnormal neutrophil myeloperoxidase from a patient with chronic myelocytic leukemia. PMID- 6282224 TI - Cathepsin M: a lysosomal proteinase with aldolase-inactivating activity. PMID- 6282223 TI - Regulation of cardiac glycogen synthase by phosphorylation: catalysis of inactivation by cAMP-dependent and cAMP-independent protein kinases and comparison with the phosphorylation of the skeletal muscle enzyme. PMID- 6282225 TI - Aging-related decrease in hepatic cytochrome oxidase of the Fischer 344 rat. PMID- 6282226 TI - [Costal micrometastasis from a cerebral astrocytoma. Immunohistochemical detection (author's transl)]. PMID- 6282227 TI - [Cancer in Madagascar. Apropos of 11,151 malignant tumors diagnosed from 1954 to 1978 in the Laboratory of Pathologic Anatomy of the Pasteur Institute]. PMID- 6282228 TI - [Influenza and lemurs]. PMID- 6282229 TI - Elevation of serum angiotensin-converting enzyme level. Occurrence in alcoholic liver disease. AB - Serum angiotensin-converting enzyme (ACE) levels were measured in 151 patients with chronic alcoholism and alcoholic liver disease. The mean serum ACE level was elevated to 30.8 +/- 13 units/mL compared with 22.8 +/- 6 units/mL in control subjects. Approximately 30.0% of the patients had elevated ACE levels. Abstinence from alcohol for six to 27 months by 11 patients was associated with persistently normal serum ACE levels. Angiotensin-converting enzyme level elevations did not correlate with abnormalities of other liver function test results or with any acute clinical condition associated with alcoholic cirrhosis. Hypoxemia was not present in the patients with elevated serum ACE levels. Elevations of serum ACE levels in patients with alcoholic liver disease may relate to an effect of alcohol on the hepatic-sinusoidal lining cells. This elevation could interfere with the use of this test for supporting the diagnosis of sarcoidosis. PMID- 6282230 TI - Captopril. PMID- 6282231 TI - [Serum concentration of C3 in primary lung cancer patients]. PMID- 6282232 TI - [Transmissible gastroenteritis in a swine-raising unit of 1,200 productive sows. 1. Clinical course, disease control and piglet loss]. PMID- 6282233 TI - [Transmissible gastroenteritis in a swine raising unit of 1,200 productive sows. 2. Results of serological follow-up studies]. PMID- 6282234 TI - [Transmissible gastroenteritis in a swine raising unit of 1,200 productive sows. 3. Course of selection-based eradication]. PMID- 6282235 TI - [Differential electron microscopic diagnosis of bovine leukosis virus and bovine syncytial virus in cell cultures]. PMID- 6282237 TI - Ruptured aortic aneurysm seen initially as acute femoral neuropathy. PMID- 6282238 TI - Complications of drainage system for modified radical mastectomy. PMID- 6282236 TI - Release of lysosomal enzymes after partial hepatectomy: study of patients with and without cirrhosis of the liver. AB - Serum activities of lysosomal enzymes beta-glucuronidase and acid phosphatase were serially estimated in 14 patients with and without cirrhosis of the liver who underwent 40% to 80% hepatic resection. Substantial increases in enzyme activities were observed two to eight weeks after operation in ten of 11 patients who did not suffer from postoperative liver failure. Regeneration of the residual livers was almost satisfactory in all 11, as evidenced by clinical, roentgenologic, and histologic findings. In three patients with advanced cirrhosis who died of hepatic failure 21 to 39 days after extensive hepatic resection, there was neither the enzymatic reaction nor evidence of regeneration of the liver remnants. In the light of this study and our previous experimental studies, serial determination of the lysosomal enzyme activities in blood is probably a beneficial biochemical index for detection of progressive hepatic regeneration. PMID- 6282239 TI - [Morphological and pathogenetic problems of cytomegaly]. AB - Cytomegalia was detected in 145 (17.6%) out of 823 fatal cases in children at autopsy. The analysis of the material suggests that the pattern of tissue reactions in this infection depends upon the immunologic responsiveness of the host entirely. In the process of maturation of the organs of immunity system, lymphohistiocytic infiltration of stroma becomes nodular, and cytomegalic metamorphosis of the cells is inhibited down to complete cessation. Histological, histochemical, and histoenzymatic methods were used to study in detail the structure of the nodular infiltrate which was found to belong to reactions of delayed type hypersensitivity. Original views concerning the mechanism of the development of cytomegalic sialoadenitis are presented. PMID- 6282240 TI - [Histogenetic and morphogenetic problems of pleomorphic adenoma of the salivary glands]. AB - Electron microscopic examinations of pleomorphic adenoma tissue showed epithelial and myoepithelial cell elements to take part in morphogenesis of both epithelial and "mesenchymal" zones. The epithelial component of pleomorphic adenoma is represented by cells showing signs of structural-functional differentiation of the squamous epithelium,, epithelium of terminal secretory parts of the salivary gland, and myoepithelial cells. The "mesenchymal" component is also represented by myoepithelial and epithelial cells. Differences in the structure of the epithelial and "mesenchymal" components are due to the pattern of arrangement and the functional status of the cells rather than to the set of the cell forms. A slow growth of the tumor and the marked functional activity of myoepithelial cells result in the accumulation of the substance of basal membranes released into the intercellular space, separation of the cells and degenerative changes in them. The results of ultrastructural examinations confirm the theory of the epithelial genesis of salivary gland pleomorphic adenoma. In this connection it sould be acknowledged that the term "mixed tumor" does not relate the essence of this neoplasia because the point at issue is a tumor of epithelial nature, whereas the term "pleomorphic adenoma" recommended by the WHO is more acceptable and emphasized the epithelial origin and heterogeneity of the cell composition and structures of the tumor. PMID- 6282241 TI - The effects of fluoridated water on rat urine and tissue cAMP levels. AB - Male Wistar rats were fed a fluoride deficient diet (less than 0.5 parts/10(6) F), and either distilled water or fluoridated water (1.0 parts/10(6)). By week 3, the control group had urinary excretions of 106 +/- 5 nmol cAMP/day (mean +/- SEM) whereas the experimental group excreted 129 +/- 6 nmol cAMP/day. After 111 days, the control group excreted 270 +/- 26 nmol cAMP/day compared to 600 +/- 78 nmol cAMP/day for the experimental group. Body weight, food and water consumption, urine volume, and urinary creatinine and phosphate levels were not significantly different between the two groups. Tissue cAMP levels were determined after 4, 6 and 16 weeks. By week 4, the rats receiving the fluoridated water had significantly higher levels of cAMP in the liver (113 per cent) tibia (130 per cent), femur (89 per cent) and heart (35 per cent). At week 6, the liver (119 per cent), tibia (296 per cent), heart (168 per cent), kidney (73 per cent) and submandibular gland (27 per cent) had significantly higher levels of cAMP. By week 16, the liver, femur, kidney and submandibular gland continued to have elevated levels of cAMP. Liver glycolytic metabolites were determined after 6 weeks, and the results suggested a decrease in pyruvate kinase activity. PMID- 6282242 TI - Cystosarcoma phyllodes: a review of 22 patients plus a case report of the youngest patient recorded in the literature. PMID- 6282243 TI - Persistence of infectious bursal disease virus in experimentally infected chickens. PMID- 6282244 TI - Jaundice in a Siamese cat with in utero feline calicivirus infection. PMID- 6282245 TI - Avian haemagglutinating adenovirus antibodies in Australian poultry. PMID- 6282246 TI - Antibody to more than one bluetongue virus serotype in Western Australia. PMID- 6282247 TI - The clinico-pathological effect of bluetongue virus serotype 20 in sheep. AB - Fifty-four Merino crossbred sheep were inoculated with bluetongue virus serotype 20 (BTV-20) by the intravenous, subcutaneous and intradermal routes. BTV-20 was successfully transmitted by Culicoides (Avaritia) spp. No. 5 to two additional sheep. Clinical and pathological effects were studied. In the artificially infected sheep, clinical signs were observed after an incubation period of 6 to 10 days and consisted of pyrexia, oral and subcutaneous hyperaemia mild oedema of the ears, face and lips, and coronitis. The major internal pathological changes were petechial and ecchymotic haemorrhages in the tunica media of the pulmonary artery near its junction with the heart and mild haemorrhage and mild oedema in the intestines, coronet, lips, cheeks and ears. Viraemia was detected between day 2 and day 14 post inoculation. The two sheep infected by insect transmission were mildly affected and became viraemic between 16 and 19 days after transmission. No deaths occurred and under experimental conditions BTV-20 caused only mild disease in housed sheep. To date there has been no reported outbreak of natural bluetongue infection in Australia. Compared to other serotypes BTV-20 appears to be of low pathogenicity in sheep. PMID- 6282248 TI - Enteritis in foals induced by rotavirus and enterotoxigenic Escherichia coli. AB - Colostrum-deprived, colostrum-fed or suckling foals were orally inoculated with foal rotavirus and enterotoxigenic Escherichia coli derived from a calf. Neither agent given alone caused diarrhoea in foals aged 1 or 2 days, although with rotavirus, 2 of the 3 inoculated foals became depressed 3 days after inoculation and all 3 were excreting rotavirus in the faeces. Inoculation of both agents induced diarrhoea in colostrum-deprived, colostrum-fed or suckling foals aged up to 16 days. There was an apparent age-related resistance to diarrhoea which developed between 2 and 3 weeks of age. It was related to failure of rotavirus to establish apparent infection in older foals and was independent of preinoculation maternal antibody. PMID- 6282249 TI - Evidence for the involvement of central alpha 2 mechanisms in intrathoracic volume expansion--induced diuresis: a study with clonidine and propranolol. AB - Intravenous (10 micrograms/kg) or intracisternal (1 microgram/kg) clonidine inhibited the diuretic response to negative pressure breathing (NPB) and left atrial distension (LAD) in chloralose anesthetized dogs. The drug reduced the induced tachycardia, but not the increase in respiratory rate caused by NPB, and did not change the blood pressure. Propranolol (1 mg/kg i.v.) did not change the NPB-induced diuresis. Intravenous yohimbine (1 mg/kg i.v.) opposed the effects of intravenous or intracisternal clonidine, whereas prazosin (0.05 mg/kg i.v.) had no effect. These results show that the adrenergic receptor implicated in the volumetric control of vasopressin secretion could be of the alpha 2 subtype. This alpha 2 adrenoceptor could be centrally located. Clonidine might therefore be proposed to combat the dehydration observed after long-term weightlessness. PMID- 6282250 TI - The influence of rendering rats anosmic on the poisoned-partner effect. PMID- 6282251 TI - Steady-state H+/O stoichiometry of liver mitochondria. AB - We have measured the H+/O stoichiometry of rat liver mitochondria respiring in a steady-state, using a novel method. This involves measuring the initial rate of H+ back-flow into mitochondria after respiratory inhibition, with the assumption that this is equal to the steady-state H+-ejection rate. Division by the steady state O2-consumption rate yields the H+/O ratio. The H+/O values obtained were: 8.3 +/- 1.0 (mean +/- S.E.M.) for 3-hydroxybutyrate: 8.2 +/- 0.7 for glutamate plus malate; 6.0 +/- 0.2 for succinate; 4.1 +/- 0.3 for ascorbate/tetramethylphenylenediamine and 3.0 +/- 0.1 for ascorbate/ferrocyanide. These values correspond to H+/O stoichiometries for electron flow to oxygen from NAD+-linked substrates, succinate and cytochrome c of 8, 6 and 2 (charge/O ratio = 4) respectively. PMID- 6282252 TI - Evidence for the extracellular reduction of ferricyanide by rat liver. A trans plasma membrane redox system. AB - 1. Reduction of ferricyanide by the isolated perfused rat liver and by isolated rat hepatocytes was studied. 2. Ferricyanide was reduced to ferrocyanide by the perfused liver at a linear rate of 0.22mumol/min per g of liver. Ferricyanide was not taken up by the liver and the perfusate concentration of ferricyanide+ferrocyanide remained constant throughout the perfusion. Perfusate samples from livers perfused without ferricyanide did not reduce ferricyanide. 3. Isolated hepatocytes reduced ferricyanide in a biphasic manner. The initial rate of 2.3mumol/min per g of cells proceeded for approx. 3min and derived from low affinity sites (apparent K(m)>1.3mm). The secondary rate of 0.29mumol/min per g of cells was maintained for the remainder of the incubation and derived from higher affinity sites (apparent K(m)0.13mm). Disruption of the cells resulted in an increase in the low-affinity rate and a decrease in the high-affinity rate. 4. Ferrocyanide was oxidized by isolated hepatocytes but not by perfused liver. The apparent K(m) for ferrocyanide oxidation by hepatocytes was 1.3mm. 5. Oxidized cytochrome c was reduced by isolated hepatocytes in the presence of 1mm-KCN but at a rate less than that of the reduction of ferricyanide. 6. Properties of the ferricyanide-reducing activities of intact hepatocytes and the perfused liver were examined. The low-affinity rate, present only in cell and broken cell preparations, was inhibited by 1mum-rotenone and 0.5mm-ferrocyanide, and stimulated by 0.1mm-KCN. The mitochondrial substrate, succinate, also stimulated this rate. The perfused liver showed only a high-affinity activity for ferricyanide reduction. This activity was also present in liver cells and was unaffected by rotenone, antimycin A, KCN, NaN(3), or p-hydroxymercuribenzoate but was inhibited by 2.6mm-CaCl(2), 2-heptyl-4-hydroxyquinoline-N-oxide and ferrocyanide. Overall, these results are consistent with the occurrence of a trans-plasma membrane redox system of liver that reduces extracellular ferricyanide to ferrocyanide. The reduction process shows properties which are similar to that of the NADH:ferricyanide oxidoreductase found in isolated liver plasma membranes but different from that of mitochondria. PMID- 6282253 TI - Quantitative measurements of the proton-motive force and its relation to steady state lactose accumulation in Escherichia coli. AB - The magnitude of delta psi (membrane potential), delta pH (pH gradient), lactose accumulation and cytoplasmic volume have been determined over a range of experimental conditions. A study of two probes of delta pH, benzoate and dimethyloxazolidene-2,4-dione (DMO), and four probes of delta psi, Rb+, K+, tetraphenylphosphonium (TPP+) and 3,3'-dipropylthiodicarbocyanine iodide, has been carried out. Benzoate and DMO are shown to be equivalent at pH values above the pK of DMO, but the latter may be less accurate below this pH. The cations TPP+ and Rb+ were found, by a number of criteria, to be equivalent, and TPP+ may be used in cells not pretreated with EDTA. These studies are an essential prerequisite to the use of TPP+ as a quantitative probe in untreated cells. PMID- 6282254 TI - The effects of partial and selective reduction in the components of the proton motive force on lactose uptake in Escherichia coli. AB - The relationship between the steady state lactose accumulation (delta plac) and the magnitude of the membrane potential (delta psi) and pH gradient (delta pH) has been studied at pHo5.5 and pHo7.5. An attempt has been made to differentiate between two possible means by which lactose accumulation may be reduced below the proton-motive force (delta p). Firstly, that delta psi and delta pH are not equivalent in driving lactose transport and secondly, that 'slip' reactions (beta galactoside exit via the carrier without a proton) may reduce accumulation. The data support the latter; however, our conclusions are tempered by the observation that the apparent stoichiometry (delta plac/delta p) increases to a value of at least 2 at values of delta p below 130 mV. PMID- 6282256 TI - Energization of the transport systems for arabinose and comparison with galactose transport in Escherichia coli. AB - 1. Strains of Escherichia coli were obtained containing either the AraE or the AraF transport system for arabinose. AraE+,AraF- strains effected energized accumulation and displayed an arabinose-evoked alkaline pH change indicative of arabinose-H+ symport. In contrast, AraE-,AraF+ strains accumulated arabinose but did not display H+ symport. 2. The ability of different sugars and their derivatives to elicit sugar-H+ symport in AraE+ strains was examined. Only L arabinose and D-fucose were good substrates, and arabinose was the only inducer. 3. Membrane vesicles prepared from an AraE+,AraF+ strain accumulated the sugar, energized most efficiently by the respiratory substrates ascorbate + phenazine methosulphate. Addition of arabinose or fucose to an anaerobic suspension of membrane vesicles caused an alkaline pH change indicative or sugar-H+ symport on the membrane-bound transport system. 4. Kinetic studies and the effects of arsenate and uncoupling agents in intact cells and membrane vesicles gave further evidence that AraE is a low-affinity membrane-bound sugar-H+ symport system and that AraF is a binding-protein-dependent high-affinity system that does not require a transmembrane protonmotive force for energization. 5. The interpretation of these results is that arabinose transport into E. coli is energized by an electrochemical gradient of protons (AraE system) or by phosphate bond energy (AraF system). 6. In batch cultures the rates of growth and carbon cell yields on arabinose were lower in AraE-,AraF+ strains than in AraE+,AraF- or AraE+,AraF+ strains. The AraF system was more susceptible to catabolite repression than was the AraE system. 7. The properties of the two transport systems for arabinose are compared with those of the genetically and biochemically distinct transport systems for galactose, GalP and MglP. It appears that AraE is analogous to GalP, and AraF to MglP. PMID- 6282255 TI - Hormonal regulation of retinoic acid-binding proteins in the mammary gland. AB - A cytosolic retinoic acid-binding (RAB) protein that sediments specifically as a 2S component on sucrose density gradients was detected in the mammary glands of virgin, pregnant and lactating rats. Mammary cytosol from pregnant rats contained significantly higher concentrations of cytosolic RAB protein than did cytosol from either virgin or lactating rats. The glands of pregnant animals exhibited increased concentration of cytosolic RAB protein during the first 5 days of pregnancy, and a steady decline was observed thereafter. The concentration of cytosolic RAB protein dropped to the value observed during lactation on the day 20 of pregnancy. Moreover, throughout lactation, low concentrations of cytosolic RAB protein were maintained. Daily treatment of virgin and lactating animals with 5 micrograms of oestradiol-17 beta for 1 week increased cytosolic RAB protein to concentrations comparable with those seen in pregnant rats. Progesterone, however, did not affect the mammary cytosolic RAB protein content of virgin rats. These results suggest hormonal involvement in the regulation of cytosolic RAB protein concentration of mammary gland during differentiation. PMID- 6282257 TI - Fast-atom-bombardment mass spectrometry. A new technique for the determination of molecular weights and amino acid sequences of peptides. AB - A detailed study of the mass spectra of peptides produced by the new technique of fast-atom bombardment is reported. Molecular weights of unmodified peptides containing up to 21 amino acids have been determined. In favourable cases, the molecular-weight determination may be made on as little as 0.1 nmol of sample. Positive-ion and negative-ion spectra are obtained with equal facility. With sample sizes in the range 2-50nmol, sequence information can be obtained in many cases. The technique represents an important contribution to structural studies on peptides, since (i) blocked peptides may be studied, (ii) no prior formation of chemical derivatives is necessary (except for distinction between lysine and glutamine), (iii) sensitivity is good, (iv) large peptides, and, in some cases, mixtures of peptides, can be studied, and (v) the technique is easy to operate, with ions being produced over a long period (5-30 min). PMID- 6282258 TI - The reaction of ornithine aminotransferase with ornithine. AB - Rat liver ornithine aminotransferase is found to exchange the pro-S hydrogen on the delta-carbon atom of ornithine exclusively, thus showing that the mammalian enzyme transfers the delta-amino group and not the alpha-amino group as has been demonstrated with the plant enzyme [Mestichelli, Gupta & Spenser (1979) J. Biol. Chem. 254, 640-647]. The enzyme also transfers the alpha-amino group of glutamate and the kinetics of the half reactions between the enzyme and both amino acids are compared. Rate and dissociation constants for both reactions are determined. PMID- 6282259 TI - Co-polymeric glycosaminoglycans in transformed cells. Transformation-dependent changes in the co-polymeric structure of heparan sulphate. AB - 1. Heparan sulphates from normal 3T3 fibroblasts are association-prone as indicated by their affinity for agarose gels substituted with cognate heparan sulphate species. Heparan sulphates from SV40-transformed or polyoma-virus transformed cells have no affinity for the same gels. 2. Heparan sulphates from the medium, the pericellular and intracellular pools of normal, SV40-transformed and polyoma-transformed 3T3 cells were separated into four subfractions (HS1-HS4) by ion-exchange chromatography. In general, HS1-HS3 were found in cell-derived heparan sulphates, whereas HS3-HS4 were present in the medium. The heparan sulphates from transformed cells were more heterogeneous and of lower charge density than those from the normal counterpart. 3. Degradations via periodate oxidation/alkaline elimination yielded the oligomers glucosamine-(hexuronate glucosamine)(n)-R with n=1-5 and a large proportion of N-sulphate groups. There was a large contribution of fragments n=4-5 from heparan sulphates of normal cells. These fragments were less common in low-sulphated heparan sulphates of transformed cells. In the case of medium-drived heparan sulphates all species had a low content of fragments n=4-5. 4. The size distribution of (glucuronate-N acetylglucosamine)(n) regions was assessed after deaminative cleavage. It was broad and ranged from n=1-10 for all heparan sulphate species. In the case of medium-derived heparan sulphates there were distinct differences between normal and transformed cells. In the latter chains the N-acetyl-rich segments were both shorter and longer than in the normal case. The shape of the disaccharide peak was consistent with a lower content of O-sulphate in the heparan sulphates from transformed cells. 5. It was concluded that heparan sulphates from medium or transformed cells exhibit the greatest structural deviation from the normal case. The finding of lower proportions of extended, iduronate/glucuronate-bearing, N sulphate-rich segments in heparan sulphates of transformed cells was particularly interesting in view of the fact that these elements have been associated with ability to self-interact. PMID- 6282260 TI - Evidence from electron-paramagnetic-resonance spectroscopy for a complex of sulphite ions with the molybdenum centre of sulphite oxidase. AB - Reduction of sulphite oxidase by sulphite at low pH values in Mes (4-morpholine ethanesulphonic acid) buffer gives rise to a new molybdenum(V) electron paramagnetic-resonance spectrum different from that obtained by photoreduction of the enzyme in the same medium. The spectrum is attributed to a sulphite complex of the enzyme, showing g-values of about 2.000, 1.972 and 1.963. The complex is analogous to that with the inhibitor phosphate in that it gives rise to no observable hyperfine coupling of Mo(V) to exchangeable protons. PMID- 6282261 TI - The catalytic chain of human complement subcomponent C1r. Purification and N terminal amino acid sequences of the major cyanogen bromide-cleavage fragments. AB - 1. The a- and b-chains of reduced and alkylated human complement subcomponent C1r were separated by high-pressure gel-permeation chromatography and isolated in good yield and in pure form. 2. CNBr cleavage of C1r b-chain yielded eight major peptides, which were purified by gel filtration and high-pressure reversed-phase chromatography. As determined from the sum of their amino acid compositions, these peptides accounted for a minimum molecular weight of 28 000, close to the value 29 100 calculated from the whole b-chain. 3. N-Terminal sequence determinations of C1r b-chain and its CNBr-cleavage peptides allowed the identification of about two-thirds of the amino acids of C1r b-chain. From our results, and on the basis of homology with other serine proteinases, an alignment of the eight CNBr-cleavage peptides from C1r b-chain is proposed. 4. The residues forming the 'charge-relay' system of the active site of serine proteinases (His 57, Asp-102 and Ser-195 in the chymotrypsinogen numbering) are found in the corresponding regions of C1r b-chain, and the amino acid sequence around these residues has been determined. 5. The N-terminal sequence of C1r b-chain has been extended to residue 60 and reveals that C1r b-chain lacks the 'histidine loop', a disulphide bond that is present in all other known serine proteinases. PMID- 6282263 TI - Purification and properties of a cross-linked complex between cytochrome c and cytochrome c peroxidase. AB - Cytochrome c (horse heart) was covalently linked to yeast cytochrome c peroxidase by using the cleavable bifunctional reagent dithiobis-succinimidyl propionate in 5 mM-sodium phosphate buffer, pH 7.0. A cross-linked complex of molecular weight 48 000 was purified in approx. 10% yield from the reaction mixture, which contained 1 mol of cytochrome c and 1 mol of cytochrome c peroxidase/mol. Of the total 40 lysine residues, four to six were blocked by the cross-linking agent. Dithiobis-succinimidylpropionate can also cross-link cytochrome c to ovalbumin, but cytochrome c peroxidase is the preferred partner for cytochrome c in a mixture of the three proteins. The cytochrome c cross-linked to the peroxidase can be rapidly reduced by free cytochrome c-557 from Crithidia oncopelti, and the equilibrium obtained can be used to calculate a mid-point oxidation-reduction potential for the cross-linked cytochrome of 243 mV. Mitochondrial NADH cytochrome c reductase will reduce the bound cytochrome only very slowly, but the rate of reduction by ascorbate at high ionic strength approaches that for free cytochrome c. Bound cytochrome c reduced by ascorbate can be re-oxidized within 10s by the associated peroxidase in the presence of equimolar H2O2. In the standard peroxidase assay the cross-linked complex shows 40% of the activity of the free peroxidase. Thus the intrinsic ability of each partner in the complex to take part in electron transfer is retained, but the stable association of the two proteins affects access of reductants. PMID- 6282262 TI - Fluid-phase interaction of C1 inhibitor (C1 Inh) and the subcomponents C1r and C1s of the first component of complement, C1. AB - Interactions between proenzymic or activated complement subcomponents of C1 and C1 Inh (C1 inhibitor) were analysed by sucrose-density-gradient ultracentrifugation and sodium dodecyl sulphate/polyacrylamide-gel electrophoresis. The interaction of C1 Inh with dimeric C1r in the presence of EDTA resulted into two bimolecular complexes accounting for a disruption of C1r. The interaction of C1 Inh with the Ca2+-dependent C1r2-C1s2 complex (8.8 S) led to an 8.5 S inhibited C1r-C1s-C1 Inh complex (1:1:2), indicating a disruption of C1r2 and of C1s2 on C1 Inh binding. The 8.5 S inhibited complex was stable in the presence of EDTA; it was also formed from a mixture of C1r, C1s and C1 Inh in the presence of EDTA or from bimolecular complexes of C1r-C1 Inh and C1s-C1 Inh. C1r II, a modified C1r molecule, deprived of a Ca2+-binding site after autoproteolysis, did not lead to an inhibited tetrameric complex on incubation with C1s and C1 Inh. These findings suggest that, when C1 Inh binds to C1r2-C1s2 complex, the intermonomer links inside C1r2 or C1s2 are weakened, whereas the non covalent Ca2+-independent interaction between C1r2 and C1s2 is strengthened. The nature of the proteinase-C1 Inh link was investigated. Hydroxylamine (1M) was able to dissociate the complexes partially (pH 7.5) or totally (pH 9.0) when the incubation was performed in denaturing conditions. An ester link between a serine residue at the active site of C1r or C1s and C1 Inh is postulated. PMID- 6282264 TI - Spectral properties of the oestrogen-induced rat uterus peroxidase II and some of its derivatives. AB - 1. The visible absorption spectrum of peroxidase II, isolated from the uterine tissue of oestradiol-treated rats, and some of its derivatives were recorded. The spectral properties of this enzyme are very similar to eosinophile peroxidase and lactoperoxidase, suggesting that these enzymes may have a similar form of haem as prosthetic group. 2. The uterine peroxidase is modified upon interaction with H2O2 and the difference spectrum of this modified enzyme is similar to that of complex II of lactoperoxidase. The modified enzyme was found to revert spontaneously to the native enzyme at rates which depended on the concentration of free enzyme and H2O2. PMID- 6282265 TI - A cytochrome c methyltransferase from Crithidia oncopelti. AB - The mitochondrial cytochrome c-557 of Crithidia oncopelti contains two lysine residues and an N-terminal proline residue that are methylated in vivo by the methyl group of methionine. The purified cytochrome can act as a methyl acceptor for a methyltransferase activity in the cell extract that uses S adenosylmethionine as methyl donor. Crithidia cytochrome c-557 is by far the best substrate for this methyltransferase of those tested, in spite of the fact that methylation sites are already almost fully occupied. The radioactive uptake of [14C]methyl groups from S-adenosylmethionine occurred only at a lysine residue ( 8) and the N-terminal proline residue. This methyltransferase appears to differ from that of Neurospora and yeast [Durban, Nochumson, Kim, Paik & Chan (1978) J. Biol. Chem. 253, 1427-1435; DiMaria, Polastro, DeLange, Kim & Paik (1979) J. Biol. Chem. 254, 4645-4652] in that lysine-72 of horse cytochrome c is a poor acceptor. Also, the Crithidia methyltransferase appears to be stable to carry lysine methylation much further to completion than do the enzymes from yeast and Neurospora, which produce very low degrees of methylation in native cytochromes c. PMID- 6282266 TI - Cytoplasmic RNA sequences complementary to cloned chick delta-crystallin cDNA show size heterogeneity. AB - Double-stranded complementary DNA (cDNA) sequences were prepared from day-old chick lens total polysomal RNA and inserted into the unique PstI restriction site of the plasmid pBR322. Colonies containing sequences complementary to abundant lens poly(A)-containing RNA sequences were identified by using lens 32P-labelled cDNA. Some of these clones have been characterized as containing delta-crystallin mRNA coding sequences by genomic DNA blot hybridization and RNA blot hybridizations. Hybridization of labelled DNA from such clones to RNA blots detected four size classes of delta-crystallin RNA sequences, although Southern blots indicated that there are probably only two delta-crystallin genes. PMID- 6282267 TI - Bis-(5'-guanosyl) tetraphosphatase in rat tissues. AB - The occurrence and distribution of bis-(5'-guanosyl) tetraphosphatase activity towards dinucleoside tetraphosphates between the 27 000 g supernatant and sedimented fraction were studied in liver, kidney, brain, muscle and intestinal mucosa from rat. The p1p4-bis-(5'-guanosyl) tetraphosphate-hydrolysing activities found in total homogenates were 0.77, 1.44, 0.39, 0.36 and 2.14 units (mumol/min)/g respectively. The activities found in the 27000 g-sedimented fractions were 74, 49, 11, 4 and 96% of those present in the homogenates respectively. The properties of the soluble enzymes were investigated. All of them have low Km values for p1p4-bis-(5'-guanosyl) tetraphosphate (from 2 to 50 microM), are competitively inhibited by guanosine 5'-tetraphosphate with K1 values from 10 to 160 nM, have molecular weights of about 21 000, require Mg2+ or Mn2+ and are inhibited by Ca2+. These properties show that bis-(5'-guanosyl) tetraphosphatase (EC 3.6.1.17), an enzyme previously characterized in Artemia salina and rat liver [Warner & Finamore (1965) Biochemistry 4, 1568-1575; Vallejo, Sillero & Sillero (1974) Biochim, Biophys. Acta 358, 117-125; Lobaton, Vallejo, Sillero & Sillero (1975) Eur. J. Biochem. 50, 495-501], is present in all the rat tissues examined. The inhibition of the enzyme by Ca2+ could be related to the effect of p1p4-bis-(5'-adenosyl) tetraphosphate as a trigger of DNA synthesis [Grummt, Waltl, Jantzen, Hamprecht, Huebscher & Kuenzle (1979) Proc. Natl. Acad. Sci. U.S.A. 76, 6081-6085]. PMID- 6282268 TI - Microvillar iron-binding glycoproteins isolated from the rabbit small intestine. AB - Rabbit intestinal microvillus membranes possess high-affinity receptors for iron whose activity reflects homeostatic changes in mucosal iron transport. To isolate and characterize these membrane components, purified microvilli were radiolabelled with 59Fe(II) and solubilized in Triton X-100. 59Fe in 105000g supernatants co-eluted with a major broad protein peak (Mr approx. 100000) on gel permeation chromatography and was rendered diffusible by Pronase digestion but not mild periodate degradation. Fluorescence studies with castor-bean lectin conjugates showed specific binding of this affinity probe exclusively to brush border membranes in the intestinal epithelium. Affinity chromatography of solubilized membrane proteins showed binding to columns of immobilized lectin. Elution with D(+)-galactose released glycoprotein-bound 59Fe purified up to sevenfold over initial membrane extracts. The lectin bound up to 82% of protein bound 59Fe. In contrast polyspecific antisera raised against rabbit microvilli in guinea-pigs precipitated less than 10% of solubilized radioactivity. Significantly more protein-bound 59Fe in detergent extracts of microvilli purified from bled animals interacted specifically with the lectin, suggesting that membrane glycoprotein receptors are involved in the homeostatic control of intestinal iron transport. PMID- 6282269 TI - Simulations of the roles of multiple cyclic nucleotide phosphodiesterases. AB - 1. Simulations were performed using a model for cellular cyclic AMP metabolism involving a hormone-activated adenylate cyclase and two cyclic nucleotide phosphodiesterases with different Michaelis constants. 2. The response curves of cyclic AMP concentration as a function of hormone concentration were affected by regulating the phosphodiesterases. The maximum velocity of the high-affinity phosphodiesterase (V1) was important in determining the position of the response curve; when v1 was less than the maximal activity of adenylate cyclase (Vc), sigmoid response curves were readily produced. The maximum attainable concentration of cyclic AMP was determined primarily by V1 when Vc less than V1, and primarily by the activity of the low-affinity enzyme when Vc greater than V1 (V2 much greater than Vc in all cases). 3. The glucagon-stimulated adenylate cyclase and insulin-stimulated phosphodiesterase of the rat liver plasma membrane were simulated using experimentally determined values for the enzyme-kinetic parameters, and a considerable potential for regulation of the system by insulin was demonstrated. 4. Other possible functions for the regulation of phosphodiesterases are considered, in particular the value of increasing the speed of response to decreases in hormone concentration. PMID- 6282270 TI - Degradation by cultured fibroblasts and macrophages of unmodified and 1,2 cyclohexanedione-modified low-density lipoprotein from normal and homozygous familial hypercholesterolaemic subjects. AB - Monolayer cultures of human skin fibroblasts and monocyte-derived macrophages were used to examine the effect of cyclohexane-1,2-dione modification on the proteolytic degradation of 125I-labelled low-density lipoprotein (LDL) from normal subjects (NLDL) and homozygous familial hypercholesterolaemic subjects (FHLDL). Normal fibroblasts, pre-incubated in lipoprotein-deficient serum, and macrophages, pre-incubated in whole serum, exhibited both saturable and non saturable degradation of LDL. In fibroblasts, the saturable receptor-mediated degradation of FHLDL was similar to that of NLDL and was abolished if the lipoproteins were modified with cyclohexanedione. The rate of non-saturable degradation of FHLDL was at least 3-fold higher than that of NLDL and each was decreased by approx. 60% after modification. In macrophages, saturable degradation was decreased but not abolished by modification. The apparent affinity for unmodified LDL was lower than that of the fibroblast receptor and was greater for NLDL than for FHLDL. Non-saturable degradation of FHLDL by macrophages was only slightly higher than that of NLDL. Modification with cyclohexanedione decreased the rate of non-saturable degradation of NLDL by 30%, but increased that of FHLDL by 75%. These experiments show differences between the degradation of 125I-labelled NLDL and FHLDL. They suggest that macrophages can degrade LDL by a saturable process with different properties from that mediated by the fibroblast receptor and that, in vitro, the rate of degradation of the modified LDL is not the same as the rate of non-receptor-mediated degradation of unmodified LDL. PMID- 6282271 TI - Lipolysis in rat adipocytes during pregnancy and lactation. The response to noradrenaline. AB - 1. Lipolysis has been measured in parametrial adipocytes from virgin, pregnant and lactating rats. 2. The basal rate and the maximal rate of lipolysis, the latter measured in the presence of noradrenaline and theophylline, remained constant between the three experimental categories, with the exception of a significant transient increase in the basal rate at parturition. 3. The noradrenaline-stimulated lipolysis rate rose above the virgin rate during pregnancy and fell below it during lactation; inclusion of adenosine deaminase in incubations abolished these differences in response to noradrenaline. 4. Cyclic AMP phosphodiesterase activity was lower in adipocytes during pregnancy and lactation than in virgin animals. PMID- 6282272 TI - The control by Ca2+ of the polyphosphoinositide phosphodiesterase and the Ca2+ pump ATPase in human erythrocytes. AB - 1. Both the Ca(2+)-pump ATPase and the polyphosphoinositide phosphodiesterase of the erythrocyte membrane can, when assayed under appropriate conditions, be activated by Ca(2+) in the micromolar range. We have therefore compared the mechanisms and affinities for Ca(2+) activation of the two enzymes in human erythrocyte membranes, to see whether the polyphosphoinositide phosphodiesterase would be active in normal healthy erythrocytes. 2. At physiological ionic strength and in the presence of calmodulin, the Ca(2+)-pump ATPase was activated by Ca(2+) in a highly co-operative manner, with half-maximal activation occurring at about 0.3mum-Ca(2+). At an optimal Ca(2+) concentration, calmodulin stimulated the Ca(2+)-sensitive ATPase activity about 10-fold. 3. Ca(2+) activated the polyphosphoinositide phosphodiesterase in a non-co-operative manner. The Ca(2+) requirements for breakdown of phosphatidylinositol 4-phosphate and phosphatidylinositol 4,5-bisphosphate were identical, which supports our previous conclusion that Ca(2+) activates a single polyphosphoinositide phosphodiesterase that degrades both lipids with equal facility. Added calmodulin did not affect the activity of the polyphosphoinositide phosphodiesterase. 4. At low ionic strength in the absence of Mg(2+), half-maximal activation of the phosphodiesterase was at about 3mum-Ca(2+). The presence of 1mm-Mg(2+) shifted the Ca(2+) activation curve to the right, as did elevation of the ionic strength. When the Ca(2+)-pump ATPase and the polyphosphoinositide phosphodiesterase were assayed in the same incubations and under conditions of intracellular ionic strength and Mg(2+) concentration, the ATPase was fully activated at 3mum-Ca(2+), whereas no polyphosphoinositide phosphodiesterase activity was detected below 100mum-Ca(2+). 5. The Ca(2+)-pump ATPase of the erythrocyte membrane normally maintains the Ca(2+) concentration of healthy erythrocytes below approx. 0.1mum. It therefore seems unlikely that the polyphosphoinositide phosphodiesterase of the erythrocyte membrane ever expresses its activity in a healthy erythrocyte. PMID- 6282273 TI - Inhibition of microsomal drug metabolism by mitochondria and cytochrome c oxidation of extramitochondrial NADPH. AB - Microsomal aniline p-hydroxylase and aminopyrine N-demethylase activities were inhibited by mitochondria. The magnitude of the inhibition increased in parallel with the amount of added mitochondria. The inhibition was reverted by 0.2 mM KCN. Marked inhibition of these microsomal enzyme activities was observed also in the presence of cytochrome c and low amounts of mitochondria causing negligible inhibition in themselves. The inhibition increased with the concentration of cytochrome c and it was reverted by KCN. Microsome-free mitochondria did not oxidize NADPH even in the presence of cytochrome c, although NADH oxidation has been demonstrated under these circumstances [Sottocasa et al., J. cell Biol. 32, 415, (1967)]. However, completion of the system by addition of microsomes resulted in the oxidation of NADPH, which was inhibited by KCN. These findings may indicate the cooperation of the microsomal and mitochondrial compartments in the regulation of drug metabolism. PMID- 6282274 TI - Gold suppression of human neutrophil function in vitro. AB - The effect of myochrysine and Auranofin on leukocyte function were measured using quantitative leukocyte iodination. Both suppressed iodination at concentrations achieved in patients. Under conditions of leukocyte submaximum stimulation, enhanced gold suppression was observed. The active portion of Myochrysine appeared to be protein bound while the active portion of Auranofin appeared to be free. Preincubation experiments indicated suppression of the myeloperoxidase halide system. Inhibition of this probable mediator of inflammation may be one of the modes of action of gold. PMID- 6282275 TI - Propranolol-induced structural changes in human erythrocyte ghost membranes. A spin label study. AB - The effect of propranolol on the structure of human erythrocyte membranes was studied using a spin labeling technique. Changes in electron spin resonance spectra of spin labeled membrane proteins were detected at concentration of the drug corresponding to its antihemolytic effect on intact erythrocytes. The character of spectral changes suggests that propranolol-induced alterations in organization of membrane proteins are connected mainly with perturbation of protein sites located on membrane surface. Propranolol also produces a decrease in order parameter of membrane lipids. The disordering effect is, however, small and detectable only at relatively high concentrations of the drug. PMID- 6282277 TI - Vanadate stimulates adenylate cyclase via the guanine nucleotide regulatory protein by a mechanism differing from that of fluoride. AB - Vanadate stimulates adenylate cyclase activity in turkey erythrocyte membranes. The maximal stimulation is 7-fold over basal at 3 mM vanadate; higher concentrations are inhibitory. A suboptimal concentration of fluoride (1 mM) together with vanadate (3 mM) activates adenylate cyclase in a non-additive manner; cyclase activation by optimal fluoride (10 mM) is inhibited by vanadate (3 mM). There is no stimulation by vanadate of adenylate cyclase activity (measured either with Mg2+ or Mn2+) in CYC- S49 lymphoma cell membranes. Vanadate (3 mM) shows no effect on binding of Beta-adrenergic agonists or antagonists to the [3H] (-)-dihydroalprenolol binding site in turkey erythrocyte membranes. These results suggest that the effect of vanadate on Adenylate cyclase is mediated through the nucleotide regulatory protein and may act by a mechanism similar to fluoride. However, in cholera toxic-treated membranes as well as in GDP-beta-S plus isoproterenol-treated membranes, fluoride-stimulated adenylate cyclase activity is significantly reduced, but vanadate stimulation is not. Our results suggest that although the actions of vanadate and fluoride in adenylate cyclase may each involve the nucleotide regulatory unit, the exact mechanisms of activation by the two anions differ. PMID- 6282276 TI - Effect of cyclooxygenase inhibitors and protease inhibitors on phorbol-induced stimulation of oxygen consumption and superoxide production by rat pulmonary macrophages. AB - Oxygen consumption and superoxide anion production by pulmonary macrophages are both increased by phorbol myristate acetate (PMA) but the two processes have been separated using protease inhibitors and cyclooxygenase inhibitors. Pretreatment with the protease inhibitors (L-1-tosylamido-2-phenylethylchloromethyl ketone (TPCK) and N-alpha-p-tosyl-L-lysine chloromethyl ketone (TLCK), as well as with the cyclooxygenase inhibitors acetylsalicylic acid (ASA) and ibuprofen (IBU), inhibited the stimulation of superoxide production and oxygen consumption by phorbol myristate acetate. However, whereas the order of potency for inhibition of stimulation of superoxide production was TPCK greater than TLCK greater than IBU greater than ASA, the order of potency for inhibition of stimulation of oxygen consumption was ASA greater than IBU greater than TPCK = TLCK. Although all four agents were effective inhibitors of PMA-stimulated superoxide production and oxygen consumption when added before PMA, in contrast to the cyclooxygenase inhibitors. TPCK was unable to inhibit oxygen consumption by more than 70-80% regardless of the concentration used, although superoxide generation could be inhibited completely. When added after PMA, ASA did not suppress either oxygen consumption or superoxide production and ibuprofen was only one-half as effective as an inhibitor. TPCK and TLCK, when added after PMA, accelerated the return to basal rates of both oxygen consumption and superoxide production. None of the four agents had any effect on basal superoxide production or oxygen consumption at the concentrations used. The data support the interpretation that both prostaglandin biosynthesis and protease activity may be associated with the activation of the superoxide-generating system of pulmonary macrophages. The consumption of molecular oxygen following stimulation of the cells with phorbol myristate acetate is not due solely to the generation of superoxide, however, since each process is inhibited with different potency by the same group of inhibitors. There appears to be a component of oxygen consumption which results from the activation of cyclooxygenase and, unlike superoxide production, cannot be completely inhibited by treatment with protease inhibitors. PMID- 6282278 TI - On the possible role played by hydrogen bonding in benzodiazepine-receptor interactions. PMID- 6282279 TI - Uptake and displacement of [3H]dihydroalprenolol, [3H]epinephrine and [3H]clonidine in isolated perfused rabbit lung. AB - Uptake and displacement of three adrenergic receptor ligands, [3H]dihydroalprenolol ([3H]DHA), [3H]epinephrine ([3H]EPI) and [3H]clonidine ([3H]CLON), were examined in isolated rabbit lungs by recirculating perfusion. Removal of [3H]DHA was the most extensive (85% uptake; 6.6 ml/min clearance), [3H]CLON removal was intermediate (50%; 3.8 ml/min), and [3H]EPI removal was the lowest (33%; 1.2 ml/min). Specific displacement of each radioligand from lung was attempted using several competing agents. Both (--)- and (+)-propranolol equally displaced [3H]DHA from lung. Phentolamine, (--)-phenylephrine and (--) epinephrine were unable to displace 10 nM [3H]EPI from lung, although the latter two agents did produce concentration-dependent increases in perfusion pressure. High concentrations of (--)-epinephrine, which produced near maximal physiological responses, inconsistently displaced 30-4- nM [3H]EPI from lung. [3H]Clonidine was displaced by unlabeled clonidine at concentrations that caused increases in perfusion pressure. Pretreatment of lungs with either 10 microM phentolamine or phenoxybenzamine did not alter the total amount of [3H]CLON displaced by clonidine, suggesting that [3H]CLON was displaced predominantly from non-specific sites, perhaps preventing detection of [3H]CLON displacement from specific (receptor) sites. Alternatively, these results may be interpreted as inhibition of uptake of each radioligand. Thus, both (--)- and (+)-propranolol interfered with [3H]DHA removal, suggesting a common mechanism for uptake and/or retention for these two beta-adrenergic receptor antagonists. Inhibition of [3H]EPI removal was observed only at high concentrations of (--)-epinephrine which indicates that pulmonary removal of epinephrine occurs through a low affinity uptake system. [3H]Clonidine removal was effectively inhibited by the same (microM) concentrations of unlabeled clonidine that produced physiological responses. Neither phentolamine nor phenoxybenzamine was able to interfere with pulmonary removal of [3H]CLON. Therefore, uptake and displacement of these adrenergic receptor radioligands showed no correlation with pharmacological effects produced by these agents in isolated perfused rabbit lung. The results are more closely associated with inhibition of removal and/or non-specific retention of the radioligands examined. PMID- 6282281 TI - Hypertension and (Na-K) ATPase activity in brain. PMID- 6282280 TI - Influence of 3-methylcholanthrene on liver nucleolar and nucleoplasmic activities of protein kinases and RNA polymerases. AB - The experiments were designed to investigate some details of the action of 3 methylcholanthrene (3-MC) on the regulation of transcription. After a single intraperitoneal dose of 3-MC a significant increase in the activities of both nucleolar and nucleoplasmic protein kinases in hepatic cells of young rats was found. The maximal stimulation took place 24 hr after the administration of 3-MC and the extent of activation was much greater in the nucleolar fraction. There is a significant elevation of the activities of both functional forms, free and template-engaged, of RNA polymerase A 24 hr after a single injection of 3-MC. Free and engaged forms of extranucleolar RNA polymerase B show a different behaviour: after 24 hr of 3-MC administration the engaged form is markedly enhanced while the activity of the free enzyme shows a significant decrease. The more moderate increase in total RNA polymerase B activity is obviously preceded by a transfer of the enzyme from 'free' to 'engaged' form. Since the enhancement of protein kinase activities was accompanied by the stimulation of nuclear RNA polymerases we suggest that both kinds of enzymes are involved in an epigenetic mechanism of the inducing action of 3-MC on cytochrome P1-450. PMID- 6282282 TI - Specific receptors for sulfonylureas in brain and in a beta-cell tumor of the rat. PMID- 6282283 TI - The role of steroids in human essential hypertension. PMID- 6282284 TI - Binding characteristics of [3H]guanfacine to rat brain alpha-adrenoceptors. Comparison with [3H]clonidine. AB - The tritium-labeled alpha-adrenoceptor agonist and antihypertensive drug guanfacine, N-amidino-2-(2,6-dichlorophenyl)-acetamide (sp. act. 24.2 Ci/mmole) was employed for a direct identification and characterization of alpha adrenoceptors in rat brain membranes. Its usefulness as a radioligand was studied in comparison with [3H]clonidine (sp. act. 26.7 Ci/mmole). The nonspecific binding of [3H]guanfacine to rat cerebral membranes was considerably more pronounced than that observed for [3H]clonidine. The specific binding of [3H]guanfacine (0.1 - 20 nM) and [3H]clonidine (0.1 - 20 nM) as defined as the excess over blanks containing (-)-norepinephrine (10 microM) was saturable. Scatchard analyses of these binding data indicated single populations of binding sites for both ligands. KC values of 3.9 ([3H]guanfacine) and 3.7 nM ([3H]clonidine) were calculated. Maximal number of specific binding sites amounted to 220 and 195 fmole/mg protein for [3H]guanfacine and [3H]guanfacine and [3H]clonidine, respectively. In case unlabeled guanfacine (1 microM) was used to characterize the specific bonding of [3H] guanfacine, KD value and maximal number of binding sites were about twice as high as determined in the presence of excess (-)-norepinephrine. The rate of association of both radioligands was rapid. Binding reached equilibrium by about 10-15 min of incubation. Half-maximal binding was attained at approximately 1-2 min. The rates of dissociation were biphasic. A rapid and a slow component were identified. The specific binding sites of [3H] guanfacine in rat brain possess the general characteristics of alpha 2-adrenoceptors. Selective antagonists of alpha 2-adrenoceptors, like yohimbine and rauwolscine strongly interfered with this binding. However, preferential blocking agents of alpha 1-adrenoceptors, such as prazosin and corynanthine, were weak competitors. The relative potency of agonists and antagonists in displacing [3H]guanfacine was identical to their effectiveness in competing for [3H]clonidine specific binding sites. It is concluded that [3H]guanfacine labels the same alpha 2-adrenoceptor population in rat brain as [3H]clonidine. However, [3H]guanfacine seems not as suitable as [3H]clonidine for routine use in the direct identification of alpha 2-adrenoceptors in view of its relatively high nonspecific binding. PMID- 6282285 TI - Inhibition of prolyl hydroxylase activity and collagen biosynthesis by the anthraquinone glycoside, P-1894B, an inhibitor produced by Streptomyces albogriseolus. AB - P-1894B, a potent prolyl hydroxylase inhibitor produced by Streptomyces albogriseolus subsp. No. 1894, inhibited about 50% of the activity of purified chick embryo prolyl hydroxylase at a concentration of 2.2 x 10(-6) M. The inhibition was noncompetitive with respect to (Pro-Pro-Gly)5 with a Ki of 1.8 x 10(-6)M. When excess amounts of ferrous ions or ascorbate were added to the reaction mixture, the inhibition was slightly reversed. P-1894B at a dose of 0.15 mg/kg reduced the hydroxylation of peptidyl proline and caused a significant inhibition of collagen biosynthesis in the uterus of the immature rat stimulated by the administration of estradiol-17 beta. PMID- 6282286 TI - Potentiation of the growth inhibitory effects of adenosine 3',5'-monophosphate analogues by homocysteine. AB - The growth inhibitory effects of N6-monobutyryl adenosine 3',5'-monophosphate (mbcAMP) and N6,O2'-dibutyryl adenosine 3',5' monophosphate (dbcAMP) towards Walker carcinoma in vitro are significantly potentiated by the addition of L homocysteine to the culture medium. This effect is not seen with L-cysteine or when exogenous cAMP or prostaglandin E2(PGE2) replace the butyrylated cyclic nucleotide. Combinations of mbcAMP or dbcAMP and L-homocysteine significantly inhibit nucleic acid methylations. Both the butyrylated cyclic nucleotides cause an elevation of the intracellular level of S-adenosyl-L-homocysteine (SAH), a potent inhibitor of S-adenosyl-L-methionine (SAM) dependent methyltransferases, and this is significantly enhanced in combination with L-homocysteine. The increase in SAH level produced by such combinations is proportional to the inhibition of methyl group incorporation into 5-methyl cytosine and 7-methyl guanine. These results suggest that L-homocysteine potentiates accumulation of SAH in the presence of mbcAMP and dbcAMP and that the resultant inhibition of methylation accounts for the enhanced growth inhibition. PMID- 6282288 TI - Fatty acid and glucose incorporation into human adipose tissue in non-insulin dependent diabetes and in insulinoma. Inverse relations with plasma triglyceride and glucose concentrations. AB - Decreased fatty acid and glucose incorporation into human adipose tissue (FIAT and GLIAT) are frequently found in primary hypertriglyceridemia (HTG) and might also contribute to the defective removal of lipoprotein triglyceride (TG) in non insulin-dependent diabetes mellitus (NIDDM). To study this possible mechanism, FIAT and GLIAT were determined in needle biopsy specimens from 14 patients with newly diagnosed NIDDM and in 14 age- and weight-matched controls. A patient with insulinoma and hyperinsulinism was also studied. FIAT and GLIAT processes were markedly reduced in patients with NIDDM that developed at the onset of maturity. Insulinoma patients, with normal plasma TG, showed FIAT-GLIAT values in the high to normal range before operation. A direct, highly significant correlation (P less than 0.001) was demonstrated between FIAT and GLIAT in diabetics, insulinoma and controls when considered together. Plasma TG and glucose concentrations were inversely related to FIAT and GLIAT. These relationships were independent of the degree of obesity. It is suggested that impaired FIAT and GLIAT might contribute to defective TG removal and HTG which are often demonstrated in NIDDM. PMID- 6282287 TI - Estrogen and glucocorticoid receptors in adult canine articular cartilage. AB - The cytoplasmic and crude nuclear fractions of adult mongrel dog articular cartilage contained estradiol- and dexamethasone-binding components which had properties of physiologic steroid receptors. The equilibrium dissociation constants averaged 0.37 nM for estradiol and 2.27 nM for dexamethasone. The concentrations estrogen receptors ranged from below 6 to 101 fmol/mg protein in the cytosols and from below 2.8 to 17.5 fmol/micrograms DNA in the nuclear fractions. Glucocorticoid receptors were detected in only 4 of 13 cytosols (range: 61.2-132 fmol/mg protein), whereas 10 of 13 nuclear fractions contained 0.8 to 46.8 femtomoles of the receptors for each microgram of DNA. There appeared to be no marked difference between the contents of either steroid receptor in female or male dog cartilage. No receptors were detected for androgen and progesterone. PMID- 6282289 TI - [The presence of 3,4-benzopyrene in plants I. Methods]. AB - Purification methods of vegetable extracts have been experienced during our researches on the possible presence of the aromatic polycyclic hydrocarbons (particularly 3,4 benzopyrene) in vegetables. The validity of different methods has been determined by IR spectrophotometric analysis using special apparatus (IR DATA-STATION). Vegetables extracts have been purified efficaciously by column chromatography, saponification and special methods (Grimmer's method). The described method eliminates, nearly entirely, the polluting substances that could alternate the chemical analysis of the vegetable extracts. PMID- 6282290 TI - [Drug-induced changes in chemotaxis under normal conditions and in rheumatoid arthritis]. AB - The effects of agents used in RA treatment, various drugs, RF, rheumatoid nodule and synovial fluid was studied on chemotaxis of PMNs. NSAIDS, corticosteroids, theophylline, colchicine, SOD, RF, rheumatoid nodule and synovial fluid were found to inhibit the chemotactic responsiveness while AMPc, GMPc, PGEI and immunodulator drugs enhanced chemotaxis. The results support the hypothesis that drugs tested may modulate chemotactic function by affecting cellular microtubules assembly and/or GMPc accumulation. PMID- 6282291 TI - [The production of superoxide anion (O2-) by polymorphonuclear cells in patients with rheumatoid arthritis]. AB - Superoxide anion production has been determined in controls and RA patients PMNs stimulated with zymosan, rheumatoid synovial membrane, nodule and synovial fluid, rheumatoid factor, aggregated gamma-globulins, Mycoplasma and Epstein-Barr virus. Patients with RA showed an increased production of O2- after incubation with zymosan and rheumatoid tissue extracts or synovial fluid in comparison to normal controls. The findings indicate that rheumatoid PMNs become activated by different stimuli to produce an excess of O2- which can contribute to chronic inflammatory process. PMID- 6282292 TI - [Effect of inflammation mediators and antirheumatic drugs on the production of superoxide anion (O2-) by polymorphonuclear cells in normals and rheumatoid arthritis]. PMID- 6282293 TI - [Functional aspects of neutrophils in full-term infants]. AB - The measure of superoxide anion (.O2-) generation was carried out in the neutrophils (PMNs) of newborns. Comparison between .O2- generation in the resting state and after stimulation with zymosan was considered. PMNs were incubated in their own plasma and in plasma from healthy adult subjects. The results demonstrate a decrease of .O2- generation (in comparison with adult PMNs) in the PMNs from newborn when they were incubated in their own plasma while the .O2- generation reached the values equal to adult's PMNs when the cells of newborns were incubated in the adult's plasma. However PMNs from some newborns demonstrated a low .O2- generation even though they were incubated in adult's plasma. The data are suggestive of a predominant, but not always exclusive, responsibility of plasma factors in the decrease .O2- generation by PMNs of full term babies. PMID- 6282294 TI - [Stimulation of cholinergic receptors and changes in cyclic nucleotide levels in the cerebral cortex of the guinea pig]. AB - The effect of Atropine and Physostigmine on 3'5' AMP and 3'5' GMP content was investigated in slices of guinea-pig cerebral cortex maintained at rest or electrically-stimulated. Atropine and Physostigmine did not modify either the basal content or the electrically-evoked increase of 3'5' AMP and 3'5' GMP. On the contrary, Betanechol 25 micro M significantly increased 3'5' GMP and 3'5' AMP content in slices kept at rest. The effect was abolished by Atropine 1,5 x 10(-7) M and d-tubocurarine 7 x 10(-6) M, respectively. PMID- 6282295 TI - [Circulating opioid peptides in chronic renal failure: acute effect of dialysis treatment]. AB - In 14 patients affected by chronic renal failure (7 males and 7 females) it has been evaluated the secretion of beta LPH, beta EP, ACTH and Cortisol in basal conditions and immediately after a dialytic treatment. For beta LPH and beta EP measurements on each thawed plasma a silic acid extraction and a successive peptides separation by a Sephadex G-75 column chromatography preceeded the two specific RIAs. Basal beta LPH plasma levels resulted significantly higher than in normal controls, while that of beta EP, ACTH and Cortisol were in the normal range. The dialytic treatment was able to increase ACTH and Cortisol plasma levels, without to modified beta LPH and beta EP plasma levels. PMID- 6282296 TI - Elevated plasma noradrenaline in response to beta-adrenoceptor stimulation in man. PMID- 6282297 TI - Prolonged converting enzyme inhibition following captopril in patients with renal insuffficiency. PMID- 6282298 TI - Clinical trial of ranitidine in the treatment of peptic ulcer. PMID- 6282300 TI - Fermentation of dietary fibre components in the rat intestinal tract. AB - 1. The fermentative breakdown of dietary fibre from various sources in the intestinal tract was studied using rat balance experiments and gas-liquid chromatograhic analysis of dietary fibre monomers in feed and faces. 2. On a basal diet with 690 g maize starch/kg but no added fibre, small but detectable amounts of polymeric glucose, rhamnose, arabinose, xylose, galactose, mannose and uronic acids, i.e. sugars occurring in dietary fibre, were excreted in faeces. 3. Dietary fibre in wheat bran was rather resistant to fermentation; 63% was recovered in the faeces. Guar gum, on the other hand, was almost completely fermented, whereas 19 and 25% of the uronic acids in low and high methoxylated pectin respectively, were excreted in faeces. The various constituents of sugar beet dietary fibre (approximately equal amounts of arabinose-based hemicellulose, pectin and non-starch glucan (cellulose)) showed quite variable availability for micro-organisms in that 6-12% of the arabinose, 17-25% of the uronic acids, and 52-58% of the cellulose were recovered in the faeces. 4. Faecal nitrogen excretion increased on addition of any one of the dietary fibre preparations studied, resulting in decreased true and apparent protein digestibility values. 5. The faecal dry weight increment was most pronounced when feeding bran and could then almost be accounted for by the remaining fibre and by protein. The less-prominent bulking effect of guar gum and pectins, that were much more extensively fermented, could be only partly explained by dietary fibre and protein. PMID- 6282301 TI - Diet and health of people with an ileostomy. 1. Dietary assessment. AB - 1. People with an ileostomy experience digestive problems with some foods. Why those foods are avoided is not known nor is it certain whether this interferes with the nutritional adequacy of their diet. 2. A detailed dietary assessment has therefore been made of thirty-seven subjects with ileostomies and a similar number of age- and sex-matched healthy controls. All food and drink eaten over 1 week was weighed and recorded. In addition. A larger group of seventy-nine ileostomy subjects and seventy matched controls answered a questionnaire designed to identify foods which upset them and which they avoided. 3. Total nutrient and energy intakes were similar in the two groups but the subjects with an ileostomy ate less dietary fibre (g/d; mean + SD: ileostomy subjects 18.0 +/- 5.9, controls 20.9 +/- 5.5; P less than 0.05) mainly due to lower fruit and vegetable intakes. Iron and vitamins A and C intakes were also less. 4. A majority of ileostomy subjects had a pattern of food intake different from the controls, taking more of their energy in the morning and less at night. A variety of food items upset more than half of them including nuts, pips, seeds, skins, onions, beetroot, lettuce, raw cabbage and carrot, peas, sweetcorn, mushrooms and dried fruit. 5. On the basis of the results it is possible to formulate general dietary advice for people with an ileostomy. PMID- 6282299 TI - Interaction of radiosensitizers and WR-2721. I. Modification of skin radioprotection. AB - We have studied the radiomodifying action in mouse skin of WR-2721 and misonidazole (MISO) when used alone or in combination. The radioprotection with WR-2721 was drug-dose dependent and highly influenced by the O2 concentration at the time of irradiation. Significant sensitizaton was observed with MISO, especially in air-breathing mice. The combination of WR-2721 and MISO produced a radiation response intermediate between the resistant and sensitive responses to either drug alone. The precise degree of sensitivity was dependent on the relative doses of protector and sensitizer. We have also studied the interaction of both drugs in terms of drug-induced lethality, which showed a clear toxic interaction. The WR-2721 LD50 was reduced by a factor of 1.4 with only 200 mg/kg of MISO. We conclude that the combination of WR-2721 and MISO shows an interaction in terms of drug toxicity and radiation response, such that the radioprotection of skin is reduced or even abolished with low doses of MISO. PMID- 6282302 TI - Physiological effects of fibre-rich types of bread. 1. The effect of dietary fibre from bread on the mineral balance of young men. AB - 1. Twelve young adult male volunteers were given a low-fibre white bread diet (9 g neutral-detergent fibre (NDF)/d) and a medium-fibre coarse-bran bread diet (22 g NDF/d), each lasting 20 d. In ad third period of 20 d the volunteers were subdivided in groups of four, consuming a high-fibre coarse-bran bread diet (35 g NDF/d), a medium-fibre fine-bran bread diet (22 g NDF/d, bran particle size less than 0.35 mm) or a wholemeal bread diet (22 g NDF/d). Retention of calcium, magnesium, iron, zinc and copper were determined during each 20 d period. 2. An increase of the amount of dietary fibre (through bran) from 9 g to 22 g NDF/d resulted in a significantly increased mineral intake, but also faecal excretion increased significantly; mineral retention remained almost constant. 3. Both intake and faecal excretion of all minerals studied, except faecal Ca, increased further (P less than 0.05) on the diet providing 35 g NDF/d; only Fe balance decreased significantly. No significant differences with respect to intake, excretion (except urinary Ca) and balance of the minerals could be detected between the coarse-bran bread and fine-bran bread diets providing 22 g NDF/d. Faecal Fe, Cu balance and Mg balance increased significantly during the wholemeal bread period compared to the coarse-bran bread diet providing 22 g NDF. 4. Serum cholesterol increased significantly, i.e. by 0.3 mmol/l, during the coarse-bran bread diet providing 22 g NDF, compared to the white-bread diet. 5. It is concluded that increasing the amount of bran in bread does not appear to affect mineral balance considerably but there seems to be an influence on mineral availability. The increased intake was accompanied by increased faecal excretion. PMID- 6282303 TI - Effects of oestrogen administration on vitamin B6 and tryptophan metabolism in the rat. AB - 1. In order to assess the effects of oestrogens on the metabolism of tryptophan and vitamin B6, ovariectomized rats have been maintained on diets providing known amounts of tryptophan, nicotinamide and vitamin B6. They received oestrone sulphate, 210 micrograms/kg body-wt per d, either incorporated in the diet for 8 weeks, or by daily intraperitoneal injection for periods of 1-3 d. 2. Oestrone sulphate administration caused a slight reduction in the concentration of pyridoxal phosphate in plasma. It had no effect on the concentration of pyridoxal phosphate in liver or kidney, the urinary excretion of 4-pyridoxic acid, the activation of erythrocyte aspartate aminotransferase (L-aspartate:2-oxo-glutarate aminotransferase, EC 2. 6. 1. 1) by incubation with added pyridoxal phosphate, or the activity of pyridoxal oxidase (aldehyde:oxygen oxido-reductase, EC 1.2.3.1) in the liver. 3. Oestrone sulphate administration caused an increase in the urinary excretion of kynurenine and a reduction in the activity of liver kynureninase (L-kynurenine hydrolase, EC 3.7.1.3). It had no effect on the urinary excretion of N1-methyl nicotinamide or the concentrations of nicotinamide nucleotides in blood, liver or kidney. 4. There was a considerable excess of the apoenzyme of kynureninase in the liver. Incubation of liver homogenates with added pyridoxal phosphate led to a 4- to 5-fold increase in activity. 5. We conclude that there is no evidence of any significant effect of oestrogens on vitamin B6. It is suggested that abnormalities of tryptophan metabolism in women receiving oestrogens, which have been widely attributed to drug-induced vitamin B6 depletion, can be accounted for by inhibition of kynureninase by oestrogen metabolites. PMID- 6282304 TI - The effect of interscapular brown adipose tissue removal on body-weight and cold response in the mouse. AB - 1. The removal of the interscapular brown adipose tissue (IBAT) led to an increase in body-weight of normal, lean mice as compared to anaesthetized controls. 2. No significant difference in food consumption could be detected between the two groups of mice over the period of the experiment. 3. Fat extraction of the whole carcasses with chloroform:methanol showed a statistically significant increase in fat content in the animals without IBAT. 4. There was no apparent failure in the operated animals to sustain core temperature when exposed to a cold stress situation (4 degrees for 24 h). 5. There was no difference in the wet weight, protein content or cytochrome oxidase content of the dorso cervical brown adipose tissue (DCBAT) between operated and control mice. This is indicative of a lack of proliferation of other brown adipose tissue sites in the operated mice in response to the removal of the IBAT. 6. It is suggested that brown adipose tissue is implicated in dietary thermogenesis in the mouse. PMID- 6282305 TI - Conformational transitions of frog heart ferricytochrome c. AB - A monomeric cytochrome c containing an intramolecular disulfide bond linking residues 20 and 102 was obtained from bullfrog hearts, as previously reported by Chan et al. [Chan, S. K., Walasek, O. F., Barlow, G. H., & Margoliask, E. (1967) Fed. Proc., Fed. Am. Soc. Exp. Biol. 26, 723]. The stability of the native globular conformation of this protein is enhanced relative to that of other cytochromes c; e.g., it is stable to 0.01 N HCl in the absence of neutral salts at 25 degrees C, and its denaturation transition in guanidine at neutral pH and 25 degrees C is centered at 3.8 M. Guanidine-denatured from cytochrome c renatures in two kinetic phases whose time constants differ by about 2 orders of magnitude as observed when using other cytochromes c. However, the absolute values for the time constants of the frog protein are notably smaller. We note that the time constants for cytochromes c are inversely related to the midpoints of their guanidine transitions, suggesting that within a homologous series of proteins the more stable the conformation the faster it folds. PMID- 6282306 TI - Fast- and slow-refolding forms of unfolded ribonuclease A differ in tyrosine fluorescence. PMID- 6282307 TI - Effects of S-adenosylhomocysteine analogues on vaccinia viral messenger ribonucleic acid synthesis and methylation. PMID- 6282308 TI - Modulation of adenylate cyclase activity by the physical state of pigeon erythrocyte membrane. 1. Parallel drug-induced changes in the bilayer fluidity and adenylate cyclase activity. AB - The fluorescence anisotropy probe perylene and the spin-labels 5-doxylsterate and 16-doxylstearate were used to estimate the order and internal microviscosity of the pigeon erythrocyte membrane upon perturbation by cationic or neutral amphipathic drugs (chlorpromazine, methochlorpromazine, tetracaine, and octanol) and an anionic drug, octanoic acid. Both methods gave identical results. The fluidity changes were found to strictly correlate with those of adenylate cyclase activity in the presence of GTP when perturbed by the drugs [Salesse, R., & Garnier, J. (1979) Biochim. Biophys. Acta 554, 102-113]. The cationic or neutral drugs, in an intermediate range of concentration, decreased the degree of organization and the internal microviscosity of the lipids together with the activity of the adenylate cyclase. At a higher concentration they reincreased them up to or higher than their initial level before the final destruction of the membrane structure and functions. This concentration effect was time dependent with tetracaine. The quaternary amine methochlorpromazine acted as chlorpromazine only on open ghosts. On intact cells, it inhibited catecholamine receptors at higher concentration and monotonously decreased the order and microviscosity, as the anionic amphipath octanoic acid did. This is taken as evidence that the inner leaflet of the bilayer is the seat for the observed multiphasic changes of viscosity and the control of adenylate cyclase and catecholamine receptors. This could stem from either a preferential intercalation or a surface effect of the amphipaths in the inner leaflet of the membrane. Since the basal activity of adenylate cyclase was not affected in the presence of drugs, it may be inferred that the enzyme holds its activity but that its stimulation is modulated by the membrane physical state. PMID- 6282309 TI - Modulation of adenylate cyclase activity by the physical state of pigeon erythrocyte membrane. 2. Fluidity-controlled coupling between the subunits of the adenylate cyclase system. AB - In the preceding paper, we described the parallel effect of amphipathic drugs on the fluidity and adenylate cyclase activity of pigeon erythrocyte membrane. This parallelism was found when the cyclase activity was assayed in the presence of a guanyl nucleotide and with Mg-ATP as the substrate after a preincubation of the membrane with the drugs [Salesse, R., Garnier, J., Leterrier, F., Daveloose, D., & Viret, J. (1982) Biochemistry (preceding paper in this tissue)]. However, when the regulatory protein (N) and the catalytic unit (E) were precoupled by GppNHp or fluoride before the action of the drugs, the cyclic AMP production was never inhibited. Thus, the drug-induced fluidization appeared to interfere with the efficiency of the activating coupling between N and E. Chlorpromazine even enhanced the cyclase activity: if the catecholamine receptor (R) repressed the cyclase activity in the absence of hormone [Rodbell, M. (1980) Nature (London) 284, 17-22], the loss of R molecules with chlorpromazine would prevent this inhibition and lead to hyperactivity of the enzyme. On the other hand, the comparison between two states of the adenylate cyclase system, (1) N and E reversibly precoupled in the presence of GTP and (2) R, N, and E precoupled in the presence of GTP plus isoproterenol, showed no difference between the activity curves at various drug concentrations: this may be interpreted as a permanent coupling of R and N. The main control exerted by fluidity on the activity of the adenylate cyclase system would thus be at the level of the activating coupling between the N subunit and the catalytic unit in pigeon erythrocyte membrane. PMID- 6282310 TI - Specificity of the catalytic interaction of human DNA polymerase beta with nucleic acid substrates. PMID- 6282311 TI - Salt-induced conformational changes in the catalytic subunit of adenosine cyclic 3',5'-phosphate dependent protein kinase. Use for establishing a connection between one sulfhydryl group and the gamma-P subsite in the ATP site of this subunit. PMID- 6282312 TI - On the protein accommodating site of the catalytic subunit of adenosine cyclic 3',5'-phosphate dependent protein kinase. PMID- 6282313 TI - Magnetic interactions in milk xanthine oxidase. AB - The relaxation behavior of the EPR signals of MoV, FAD semiquinone, and the reduced Fe/S I center was measured in the presence and absence of other paramagnetic centers in milk xanthine oxidase. Specific pairs of prosthetic groups were rendered paramagnetic by poising the native enzyme or its desulfo glycol inhibited derivative at appropriate potentials and pH values. Magnetic interactions were found between the following species: Mo--Fe/S I (100-fold increase in microwave power required to saturate the MoV EPR signal at 103 K when Fe/S I is reduced as opposed to oxidized), FAD--Fe/S I and FAD--Fe/S II (70-fold increase in power required to saturate the FADH.EPR signal at 173 K when either Fe/S center is reduced), and Fe/S I--Fe/S II (2.5-fold increase in power to saturate the reduced Fe/S I EPR signal at 20 K when Fe/S II is reduced). The Mo- Fe/S I interaction was also detected as a reduced Fe/S I induced splitting of the MoV EPR spectrum at 30 K. No splittings of the FADH. or Fe/S center spectra were detected. No magnetic interactions were found between FAD and Mo or between Mo and Fe/S II. These results, together with those of Coffman & Buettner [Coffman, R. E., & Buettner, G. R. (1979) J. Phys. Chem. 83, 2392-2400], were used to estimate the following approximate distances between the electron carrying prosthetic groups of milk xamthine oxidase: Mo--Fe/S I, 11 +/- 3 A; Fe/S I-Fe/S II, 15 +/- 4 A; FAD-Fe/S I, 16 +/- 4 A; FAD-Fe/S II, 16 +/- 4 A. A model for the arrangement of these groups within the xanthine oxidase molecule is suggested. PMID- 6282314 TI - Determination of the stoichiometry of electron uptake and the midpoint reduction potentials of milk xanthine oxidase at 25 degrees C by microcoulometry. PMID- 6282315 TI - Lateral diffusion of epidermal growth factor complexed to its surface receptors does not account for the thermal sensitivity of patch formation and endocytosis. AB - The patching and endocytosis of EGF (epidermal growth factor) bound to A-431 cells (a human epidermoid carcinoma line) are temperature-sensitive processes which are completely inhibited at 4 degrees C. Receptor-mediated endocytosis generally occurs through coated regions, and EGF bound to its membrane receptor must diffuse laterally to these points of internalization. In this work we investigated the thermal sensitivity of the lateral diffusion of EGF receptor complexes and the thermal sensitivity of the patching and endocytosis of the hormone receptor complexes. Using the fluorescence photobleach recovery technique, we measured the lateral diffusion coefficients of a fluorescent derivative of EGF as a function of temperature. The lateral diffusion coefficient (D) increased gradually from 2.8 X 10(-10) cm2/s at 5 degrees C to 8.5 X 10(-10) cm2/s at 37 degrees C, and no phase transition was detected. Neither was a phase transition detected when we measured the diffusion coefficient of fluorescent lipid probes over this temperature range. From a calculation of the collision frequency of the occupied EGF receptors with coated regions using our measured values of D at 5 and 37 degrees C, we conclude that diffusion is not the rate limiting step for either endocytosis or patching. PMID- 6282316 TI - Purification and properties of a presynaptically acting neurotoxin, mandaratoxin, from hornet (Vespa mandarinia). AB - A hornet (Vespa mandarinia) neurotoxin, mandaratoxin (MDTX), was purified by simple procedures with column chromatography made on Sephadex G-50 and CM Sephadex by using an acetate buffer. The molecular weight of homogeneous MDTX was calculated to be approximately 20000 by gel filtration, NaDodSO4 disc gel electrophoresis, and amino acid analysis. MDTX is a single-chain polypeptide. MDTX did not migrate electrophoretically in a basic buffer at pH 8.3 but did so when the buffer was acidic, at pH 4.3. The isoelectric point of the toxin was determined at 9.1 by isoelectric focusing. A relatively high amount of lysine was found in the amino acid analysis. A280nm1% was 15.1. Glucosamine and galactosamine were not detectable by amino acid analysis. MDTX had neither hemolytic nor enzymatic activity. The toxin was heat labile. By use of neuromuscular junctions of a lobster walking leg, it was found that the nanomole range of MDTX irreversibly blocked the excitatory postsynaptic potential without appreciable change in the resting conductance of the postsynaptic membrane. Intracellular recording from the presynaptic nerve fiber showed that MDTX blocked the action potential mainly by reducing the sodium current. PMID- 6282317 TI - Multiple activities on phosphorylase kinase. 1. Characterization of three partial activities by their response to calcium ion, magnesium ion, pH, and ammonium chloride and effect of activation by phosphorylation and proteolysis. PMID- 6282318 TI - Histone acetylation and the deoxyribonuclease I sensitivity of the Tetrahymena ribosomal gene. AB - Under appropriate conditions, up to 8.5% of the total acetate can be removed from the histones of isolated Tetrahymena macronuclei by an endogenous histone deacetylase activity. After in vitro deacetylation, the ribosomal genes are still preferentially digested by DNase I. These observations suggested that either the majority of histone-bound acetate is unnecessary to maintain the DNase I sensitive state or ribosomal chromatin (rChromatin) histones remain acetylated under these conditions. The characteristics of histones acetylation were studied in Tetrahymena rChromatin, which can be isolated in a relatively pure form. Histones associated with the presumably active, DNase I sensitive ribosomal genes have a high steady-state level of histone acetylation which, surprisingly, is maintained by very low acetate turnover rates. PMID- 6282319 TI - Formation of a polymethylene bis(disulfide) intersubunit cross-link between cysteine-281 residues in rabbit muscle glyceraldehyde-3-phosphate dehydrogenase using octamethylene bis(methane[35S]thiosulfonate). AB - The synthesis of a radioactive cross-linking agent, S,S'-octamethylene bis(methane[35S]thiosulfonate) (OBMTS), is described. The route of synthesis can be generally used in the synthesis of 35S-labeled thiosulfonates for the selective modification of thiols in proteins. Glyceraldehyde-3-phosphate dehydrogenase (G3PD) reacts asymmetrically with the bifunctional inhibitor. Initially two molecules of OBMTS react rapidly with the active-site thiol, Cys 149, on two of the four subunits to inhibit the enzyme completely without cross linking. This is followed by the modification of four Cys-281 residues to incorporate two cross-links into the tetramer. Reduction of modified G3PD with 5 mM dithioerythritol under nondenaturing conditions released the inhibitor blocking the active-site thiol and completely restored enzyme activity while leaving the cross-link intact. Sodium dodecyl sulfate (Na-DodSO4) gel electrophoresis of the cross-linked enzyme under nonreducing conditions showed a dimer (Mr 72000) as the major species which was only cleaved by reduction in Na DodSO4 containing beta-mercaptoethanol. The monomer formed was still radioactive, showing that the first disulfide in the cross-link was reduced at a much faster rate than the second disulfide. The latter was only reduced by using vigorous conditions. The location of the intersubunit cross-linked residues was established by isolation of the cyanogen bromide and tryptic subdigest peptides containing modified Cys-281. There were identified by molecular weight, amino terminal sequence, and amino acid composition. PMID- 6282320 TI - Poly(adenosinediphosphoribose) polymerase inhibitors stimulate unscheduled deoxyribonucleic acid synthesis in normal human lymphocytes. PMID- 6282321 TI - Spin-labeled pepstatin binding to pepsin. A study by electron spin resonance and nuclear magnetic resonance. PMID- 6282322 TI - Direct assignment of the dihydrouridine-helix imino proton resonances in transfer ribonucleic acid nuclear magnetic resonance spectra by means of the nuclear Overhauser effect. AB - The NMR resonances from the hydrogen-bonded ring NH protons in the dihydrouridine stem of Escherichia colt tRNA1Val have been assigned by experiments involving the nuclear Overhauser effect (NOE) between adjacent base pairs. Irradiation of the 8 14 tertiary resonance produced a NOE to base pair 13. Irradiation of the CG13 ring NH produced NOEs to base pairs 12 and 14. Similarly, base pair 12 was shown to be dipolar coupled to 11 and 13, and base pair 11 was found to be coupled to 10 and 12. These sequential connectivities led to the assignment of CG13 at 13.05 ppm, UA12 at -13.84 ppm, CG11 at -12.23 ppm, and GC10 at -12.60 ppm. The results are compared with previous, less direct assignments for these four base pairs and with the expected proton positions from the crystal structure coordinates for this helix. PMID- 6282324 TI - Kinetics of sodium ion induced calcium ion release in calcium ion loaded cardiac sarcolemmal vesicles: determination of initial velocities by stopped-flow spectrophotometry. PMID- 6282323 TI - Spectrophotometric analysis of the interaction between cytochrome b5 and cytochrome c. AB - The interaction between cytochrome c and the tryptic fragment of cytochrome b5 has been found to produce a difference spectrum in the Soret region with a maximum absorbance at 416 nm. The intensity of this difference has been used to determine the stoichiometry of complex formation and the stability of the complex formed. At pH 7.0 [25 degrees C (phosphate), mu = 0.01 M], the two proteins were found to form a 1:1 complex with an association constant, KA, of 8(3) x 10(4) M 1. The stability of the complex was found to be strongly dependent on ionic strength with KA increasing to 4(3) x 10(6) M-1 at mu = 0.001 M [25 degrees C, pH 7.0 (phosphate)]. Analysis of the dependence of KA on pH from pH 6.5 to 8 demonstrated that this complex is maximally stable between pH 7 and 8 or about midway between the isoelectric points of the two proteins. Analysis of the temperature dependence of KA revealed that formation of the complex between the two proteins is largely entropic in origin with delta Ho = 1 +/- 3 kcal/mol and delta So = 33 +/- 11 eu [pH 7.0 (phosphate), mu = 0.001 M]. This result may be explained either by the model of Clothia and Janin [Clothia, C., & Janin, J. (1975) Nature (London) 256, 705] in terms of extensive solvent reorganization upon complexation or by the model of Ross and Subramanian [Ross, P. D., & Subramanian, S. (1981) Biochemistry 20, 3096] in which the negative enthalpic and entropic contributions of short-range protein-protein interactions are offset by proton release. PMID- 6282325 TI - Involvement of arginine residues in the allosteric activation of Escherichia coli ADP-glucose synthetase. AB - Inactivation of Escherichia coli ADP-glucose synthetase (EC 2.7.2.27) by the arginine-specific reagents cyclohexanedione and phenylglyoxal resulted primarily from interference with normal allosteric activation. Partial modification by phenylglyoxal resulted in a lessened ability of fructose 1,6-bisphosphate (fructose-P2) to stimulate and of 5'-AMP (5'-adenylate) to inhibit enzymic activity. The apparent affinity for fructose-P2 and the Vmax at saturating fructose-P2 concentrations were decreased by the arginine modification. Fructose P2, 5'-adenylate, and several other allosteric effectors were able to partially protect the enzyme from inactivation. However, catalytic activity was not decreased by arginine modification under conditions where the enzyme was assayed in the absence of fructose-P2. The two arginine-modifying reagents differed markedly in their reactivity with the enzyme. Cyclohexanedione inactivated the enzyme quite slowly and eventually reacted with at least 14 of the 32 arginines present per subunit. Phenylglyoxal was some 50-fold more effective in inactivation, but it modified only one arginine residue per subunit. PMID- 6282326 TI - Effects of cholesterol evulsion on susceptibility to perfringolysin O of human erythrocytes. AB - Human erythrocytes preincubated with a phosphatidylcholine suspension (preincubated cells) showed decreased susceptibility to perfringolysin O, the decrease being strongly affected by preincubation time and temperature, and the phosphatidyl choline concentration. The binding of the toxin to the preincubated cells also decreased with the preincubation time and reached minimum at 37 degrees C for 6 h. Through this preincubation, about 30% of cholesterol was removed from cells without lysis. The susceptibility of preincubated cells to the toxin seemed to be affected by the amount of cholesterol removed from cells, but not by the cholesterol content of cell membranes. This indicates that most of the cholesterol interactive with the toxin is removable from cell membranes by preincubation with phosphatidylcholine suspension, and that the residual cholesterol is firmly constituted in the membrane structure and cannot interact with the toxin. After cholesterol evulsion by the preincubated plasma method (Murphy, J.R. (1962) J. Lab. Clin. Med. 60, 86-109 and 60, 571-578), cells also exhibited lower susceptibility to the toxin and to saponins, but higher susceptibility to lysophosphatidylcholine. PMID- 6282327 TI - Demonstration of human platelet beta-adrenergic receptors using 125I-labeled cyanopindolol and 125I-labeled hydroxybenzylpindolol. AB - The radioiodinated pindolol analogs 125I-labeled cyanopindolol ([125I]CYP) and 125I-labeled hydroxybenzylpindolol ([125I]HBP) have been used to study binding to human platelet beta-adrenergic receptors. [125I]CYP binds to a saturable class of binding sites on platelet membranes with a dissociation constant (Kd) of 14 +/- 3 pM and maximal binding capacity (Bmax) of 18 +/- 4 fmol/mg protein. Binding of [125I]CYP is reversible and is characterized by forward and reverse rate constants of 1.8 . 10(7) s-1 . M-1 and 3.8 . 10(-4) s-1, respectively. [125I]HBP binds to a saturable class of platelet membrane sites with a Kd of 50 +/- 10 pM and Bmax of 32 +/- 6 fmol/mg protein. [125I]HBP also binds to a saturable class of sites on intact platelets with a Kd of 58 +/- 14 pM and Bmax of 24 +/- 4 molecules per platelet. Binding of [125I]CYP and [125I]HBP is stereospecifically inhibited by propranolol and epinephrine; the (-) stereoisomers are at least 50 times more potent than the (+) stereoisomers. Binding of both radioligands is inhibited by adrenergic ligands with a potency order of propranolol much greater than isoproterenol greater than epinephrine greater than practolol greater than norepinephrine greater than phenylephrine. These observations indicate that [125I]CYP and [125I]HBP bind to platelet sites which have the pharmacological characteristics of beta-adrenergic receptors but which are not typical of either the beta 1 or beta 2 sub-type. PMID- 6282328 TI - Age-dependent changes in the number of [3H]ouabain-binding sites in rat soleus muscle. AB - The influence of age on the number of (Na+K+)-ATPase units in skeletal muscle has been assessed by measurements of [3H]ouabain binding in vitro and in vivo to rat soleus muscle. In vitro measurements showed that from the 2nd to the 28th day of life, the number of [3H]ouabain-binding sites increases from 120 to 580 pmol/g wet wt. This is followed by a decrease, until a plateau between 150 and 200 pmol/g is reached around 150 days after birth. 60 min after intraperitoneal injection of [3H]ouabain (12.5 mumol/kg body weight), the soleus muscles of 28 day-old rats had accumulated 2.4-times more 3H-activity per g wet wt. than muscles of 85-day-old rats and the 3H-activity in plasma was 54% lower. The results may explain the low sensitivity to digitalis glycosides found in infants as compared to premature or adult individuals. PMID- 6282329 TI - A comparison of the phage T4 gene 32 protein and Escherichia coli RNA polymerase binding sites on hamster papovavirus DNA. AB - Phage T4 gene 32 protein and Escherichia coli RNA polymerase were bound to hamster papovavirus DNA. The binding regions were identified by electron microscopy employing a protein-free spreading technique. After gene 32 protein treatment four denaturation regions could be mapped, at 0.04-0.12, 0.30-0.36, 0.50-0.60 and 0.75-0.90 DNA map units, respectively, using the unique BamHI cleavage site as zero point. Eight RNA polymerase binding sites can be found which are localized at positions 0.05; 0.11; 0.18; 0.31; 0.57; 0.66; 0.76 and 0.82. A comparison of the RNA polymerase binding sites with the gene 32 protein denaturation pattern reveals a correspondence of six of eight polymerase binding sites with (A+T)-rich regions within the hamster papovavirus genome. PMID- 6282330 TI - Phosphorylation of ribosomal proteins S3, L1 and L24 during spherulation in Physarum polycephalum. AB - The phosphate content of ribosomal proteins S3, L1 and L24 has been determined in the course of spherulation of Physarum polycephalum. The major phosphoprotein, S3, was completely dephosphorylated after 4 h of differentiation. The phosphate content of L1 and L24 was not altered during the differentiation. The cellular level of ATP remained constant for at least 5 h. A 3-fold reduction of cyclic AMP concentration occurred in the first hour, followed by a slow increase to a final value of twice the level observed in growing cells. The results showed that the phosphorylation of ribosomal proteins is regulated by at least two different mechanisms and that the dephosphorylation of S3 is not induced by a lack of cellular ATP. Although cyclic AMP might trigger the dephosphorylation of S3, the phosphate content of this protein remained at a very low value even when the cellular concentration of cyclic AMP rose significantly. Since the polysome level remains constant during the first 24 h of spherulation, the phosphorylation of S3 is not necessary for active protein synthesis and the phosphorylation of L1 and L24 is not involved in ribosome inactivation, which occurs after 24 h. PMID- 6282331 TI - Construction of gapped circular DNA from phage M13 by in vitro hybridization. PMID- 6282332 TI - DNA methylation in mouse cells in culture as measured by restriction enzymes. AB - Methylation of DNA in normal mouse cultured 3T3 cells and in their virally or chemically transformed derivatives was studied. DNA methylation was studied by restriction with HpaII, MspI, or HpaII plus MspI. DNA from the chemically transformed cells was cleaved about twice as often with HpaII than was the DNA of normal and virally transformed cells. Digests with MspI and HpaII plus MspI were identical in all cell lines studied. Densitometry of the restriction patterns allowed an estimate of total DNA methylation from the weight average lengths. The chemically transformed cell line showed 25% reduction in methylation compared to the other cell lines. Southern blot hybridization using satellite DNA showed that these sequences followed a pattern of modification similar to that of total DNA. PMID- 6282333 TI - Eukaryotic DNA repair is blocked at different steps by inhibitors of DNA topoisomerases and of DNA polymerases alpha and beta. PMID- 6282334 TI - Mechanism of autocatalytic oxidation of oxyhemoglobin by nitrite. An intermediate detected by electron spin resonance. AB - Oxidation of oxyhemoglobin by nitrite is characterized by the presence of a lag phase followed by the autocatalysis. Just before the autocatalysis begins, an asymmetric ESR signal is detected which is similar to that of the methemoglobin radical generated from methemoglobin and H2O2 in shape, g value (2.005), peak-to peak width (18 G) and other properties, except the difference in the dependence on temperature. Generation of H2O2 is indicated by the prolongation of the lag phase by the addition of catalase. On the other hand, the oxidation is modified by neither superoxide dismutase nor Nitroblue tetrazolium. The oxidation is prolonged in the presence of KCN. The present results indicate a free-radical mechanism for the oxidation in which the asymmetric radical catalyzes the formation of NO2 from NO2- by a peroxidase action and NO2 oxidizes oxyhemoglobin in the autocatalytic phase. PMID- 6282335 TI - Enhancement of hormone action by a phorbol ester and anti-tubulin alkaloids involves different mechanisms. AB - The tumor-promoting phorbol ester 12-O-tetradecanoyl phorbol-13-acetate (TPA) enhanced 1-isoproterenol and prostaglandin E1 stimulated cyclic AMP formation in clones of mouse myeloid leukemic cells. The enhancement was found up to 3h after TPA treatment and had disappeared after 24h, indicating its reversibility. The effect of TPA was not inhibited by removal of extracellular Ca2+ or pre-treatment with the calcium ionophore A23187. This enhancement by TPA seems to involve a different pathway than enhancement of response to the same hormones after treatment with the anti-tubulin alkaloids colchicine or vinblastine, since a myeloid leukemic cell mutant clone that was non-responsive to the anti-tubulin alkaloids responded to TPA. Furthermore, combined treatment of colchicine sensitive cells with TPA and colchicine showed an additive stimulating effect. The enhancement of cell response to hormones by TPA was found in myeloid leukemic cell clones that either were or were not induced to differentiate after treatment with TPA. This suggests that enhancement of the effect of these and possibly other hormones by TPA may be an initial step of TPA action, but that this enhancement is not sufficient to induce the wide repertoire of TPA effects including induction of differentiation. PMID- 6282336 TI - On the mechanism of isoproterenol-induced desensitization of adenylate cyclase in cultured differentiated hepatocytes. AB - The adenylate cyclase of cultured differentiated RL-PR-C hepatocytes is desensitized to 1-isoproterenol by exposure to this beta-agonist. Virtually complete desensitization occurred by 60 min (intact cells) or 30 min (isolated plasma membranes). Isoproterenol was maximally effective at 10 micrometers, although substantial desensitization occurred at isoproterenol concentrations as low as 10 nM. Protein synthesis was not required for desensitization. Recovery from desensitization under tissue culture conditions was only 25% complete by 24 h. Maximum desensitization was accompanied by only a modest 35% decrease in binding sites (as determined by binding assays with [3H]dihydroalprenolol), with no change in binding affinity. Adenylate cyclase desensitized to 1-isoproterenol responded normally to guanine nucleotides and to fluoride, suggesting that the regulatory and catalytic proteins were not the sites of the desensitization "defect'. Using N-ethylmaleiimide to inactive the regulatory and catalytic proteins, and dicyclohexylcarbodiimide to inactivate the beta-adrenergic receptor, of intact hepatocytes, various heterologous cell fusion hybrids were produced, and their adenylate cyclases tested for responsiveness to 1 isoproterenol; only hybrids containing "desensitized' receptor failed to respond to isoproterenol. These results suggest that the mechanism of desensitization to isoproterenol involves only the receptor component of the receptor-regulatory protein(s)-adenylate cyclase complex, and that the receptors are reduced in number and/or ability to interact with the regulatory protein as a result of the desensitization process. PMID- 6282337 TI - Study of storage iron in cultured chick embryo fibroblasts and rat glioma cells, using Mossbauer spectroscopy. AB - 57Fe Mossbauer spectra of normal and Rous sarcoma virus-infected cultured chick embryo fibroblasts and rat glioma cells have been measured between 0.08 and 318 K. Ferritin-like iron and bacterio-ferritin-like iron have been found in these cells, in various relative amounts, indicating a close relationship between the two storage materials. The bacterio-ferritin-like iron was found to be predominantly membrane-bound. Above 260 K very wide lines were observed in the Mossbauer spectra, yielding an effective viscosity of about 1 poise in the normal chick embryo fibroblasts and about 0.5 poise in the virus-infected chick embryo fibroblasts. PMID- 6282338 TI - Lack of tropomyosin correlates with the absence of stress fibers in transformed rat kidney cells. AB - We have utilized epithelial rat kidney cells and their Kirsten viral transformant (442) to examine the role of actin-binding proteins in cellular morphogenesis. Normal rat kidney cells are well spread while the transformed cells are more spherical, poorly adherent, and lack actin stress fibers (Rubin, R.W., Warren, R.H., Lukeman, D.S. and Clements, E. (1978) J. Cell Biol. 78, 28-35). By immunofluorescence, antitropomyosin prominently stains normal rat kidney cell stress fibers while only a weak, nonspecific fluorescence is observed in 442 cells. Using two-dimensional gel electrophoresis, tropomyosin can be detected in normal rat kidney cells homogenates. The tropomyosin subunits are enriched in Triton-extracted filamentous normal rat kidney cell models, and in extracts of normal rat kidney cell homogenate produced by using a rapid myosin affinity technique to isolate actin and actin-associated proteins. The identity of the tropomyosin subunits has been confirmed by electrophoretic mobility, lack of proline, and the peptide map generated by limited proteolysis. None of these techniques have detected tropomyosin in the corresponding 442 preparations. Our results suggest that the transformation of normal rat kidney cells has led to an overall reduction in tropomyosin content. This may be related to the inability of 442 cells to organize filamentous actin stress fibers. PMID- 6282340 TI - [ATP-synthetase activity, respiration and cytochromes of rat heart mitochondria in aging and hyperthyroidism]. AB - The ATP-synthetase activity, the rate of oxygen uptake under different metabolic conditions, the tightness of coupling of respiration to oxidative phosphorylation and the cytochrome contents in heart mitochondria of rats from different age groups were studied under normal conditions and in hyperthyroidism. It was found that heart mitochondria of aged animals did not practically differ in terms of their functional activity from those of the young animals. Administration of thyroxin to the animals from all age groups produced no significant effects on the state of mitochondria, increasing the rate of ATP synthesis on alpha glycerophosphate, which was especially well-pronounced in aged animals, and the cytochrome content in 1-month-old rats. PMID- 6282341 TI - [Non-specific acid nucleoside triphosphatase from cytosol and chromatin of rat liver: partial purification and general properties]. AB - The nucleoside triphosphatase (EC 3.6.1.15) was isolated from rat liver cytosol and purified 600-fold. The enzyme hydrolyzes all NTPs and dNTPs, splits NDPs and dNDPs at a low rate and does not destroy NMPs and dNMPs. The phosphatase consists of a single subunit with molecular weight of 65 000. The chromatin fraction of the enzyme is fully bound to the membrane, whereas the cytosol fraction contains 15-30% of the membrane-bound enzyme. Both free and membrane-bound phosphatases possess identical functional properties. The enzymatic activity has a pH-optimum of 4.0--4.5, is increased in the presence of Me2+ and is unaffected by ouabain, Triton X-100, N-ethylmaleimide, NaF or DNA, but is inhibited by NaCl, Pi and PPi. The value of Km is equal to 20 microM for TTP splitting. Since the NTP pool is essentially changed throughout the cell cycle, it is suggested that the nucleoside triphosphatase can participate in the nucleotide pool regulation. PMID- 6282342 TI - [Isolation, purification and properties of restriction endonuclease EcoRII]. AB - The restriction endonuclease Eco RII was isolated and purified to homogeneity. The isolation procedure involved the use of the E. coli strain B834/pSK323, containing the recombinant plasmide pSK323 which provides for the oversynthesis of Eco RII enzymes. Data from gel filtration and Na-DS electrophoresis suggest that the restriction endonuclease Eco RII is a protein made up of two subunits, each with molecular weight of 44 000. PMID- 6282339 TI - The effect of various stimuli and calcium antagonists on the fluorescence response of chlorotetracycline-loaded human neutrophils. AB - Chlorotetracycline has been used in neutrophils and other cells as probe of the state of membrane-bound calcium. We report here that human neutrophils treated with chlorotetracycline response to soluble secretagogues by a prompt decrease in chlorotetracycline fluorescence. This response was observed within 2-5 s, making it one of the most immediate reactions in neutrophils to stimulation, and was obtained with three secretagogues studied: a chemotactic peptide N-formyl methionyl-leucyl-phenylalanine, a tumor promotor (phorbol myristate acetate) and a lectin (concanavalin A). The responses of neutrophils to the three stimuli differed both quantitatively and qualitatively. The calcium EGTA, did not effect the onset of the decrease in chlorotetracycline fluorescence, suggesting that the probe was measuring changes in intracellular calcium pools. The intracellular calcium antagonists, TMb-8, W-7 and trifluoperazine, did not block, but actually augmented, the fluorescence response. All four of these calcium antagonists blocked the recovery of chlorotetracycline fluorescence which was usually observed several minutes after stimulation with N-formyl-methionyl-leucyl phenylalanine. This suggests that recovery was dependent upon both extracellular calcium and active calmodulin. The results are consistent with the hypothesis that changes in chlorotetracycline fluorescence reflect changes in a pool of membrane-bound 'trigger calcium', the release of which is an essential first step in stimulus-response coupling in human neutrophils. PMID- 6282343 TI - [Effect of freezing-thawing rates on the functional state and ionic permeability of rat liver mitochondria]. AB - The effects of various rats of freezing-thawing reactions on the functional state and ionic permeability of rat liver mitochondria were studied. The degree of mitochondrial damage during the freezing -- thawing process depended on the rate of thawing rather than on that of freezing. The mitochondria which were slowly or rapidly frozen down to --196 degrees and subsequently slowly thawed revealed a higher membrane permeability for K+ Na+ and H+ and a more than 2-fold increase of the ATPase activity and the maximal rate of NADH oxidation via the antimycin insensitive pathway in the presence of cytochrome c. This was concomitant with a complete inhibition of the ATP-synthetase activity and a marked inhibition of the respiratory chain function due to the efflux of cytochrome c from the inner mitochondrial membrane. After freezing and rapid thawing the functional activity of mitochondria changed insignificantly. A comparison of different cryoeffects demonstrated that the minimal damaging effects were exerted by rapid freezing -- rapid thawing, when the mitochondria partly restored their ability for oxidative phosphorylation. PMID- 6282344 TI - [Use of affinity chromatography for the purification of specific endonuclease Eco RI and Bg1 II]. AB - The highly active preparations of specific endonucleases Eco RI and Bgl II were purified by affinity chromatography from E. coli and Bacillus globiggii cells, respectively. The isolation and purification procedures included cell disruption by ultrasonication, ultracentrifugation and chromatography. Blue dextrane Sepharose, folate-Sepharose and phenyl-Sepharose were used as affinity adsorbents. The optimal conditions for the adsorption and elution of the endonucleases excluding intermediate steps of dialysis and concentration were selected. A high degree of purification was achieved by a consecutive use of adsorbents with different ligands. The purified enzyme does not contain non specific nucleases or phosphatases, is sufficiently concentrated and can be used for specific hydrolysis of DNA. PMID- 6282345 TI - [Characteristics of the biochemical mechanisms of electromechanical coupling in the myocardium]. PMID- 6282346 TI - Lithium does not prevent agonist-induced subsensitivity of human adenylate cyclase. AB - In a previous study 11 depressed patients were treated with salbutamol, a beta-2 adrenergic agonist, and beta-2 adrenergic receptor sensitivity was evaluated by measuring the plasma cyclic AMP rise after an iv dose of salbutamol. Salbutamol treatment induced subsensitivity of the beta-adrenergic adenylate cyclase with a time course paralleling the antidepressant effects. In the present study nine patients who were depressed despite treatment with lithium were treated with salbutamol plus lithium. Subsensitivity of the beta-adrenergic adenylate cyclase developed in the presence of lithium to the same degree as in patients treated with salbutamol alone. These results represent the first human study of the theory that lithium stabilizes receptor sensitivity changes. Lithium's failure to prevent subsensitivity agrees with reports that lithium fails to prevent impramine-induced subsensitivity of beta-adrenergic receptors in rat cortex. Lithium stabilization of receptor sensitivity of beta-adrenergic receptors in rat cortex. Lithium stabilization of receptor sensitivity would therefore appear to be unidirectional, preventing supersensitivity but not subsensitivity. PMID- 6282347 TI - Receptors and the mechanism of action of ECT. PMID- 6282348 TI - On the correlated nature of evoked brain activity: biophysical and statistical considerations. AB - The contribution of EP research to the mind-brain problem is considered. The view is promoted that the scalp EP represents a macroscopic rescaling of patterns of activity in cortical columns, which are held by many workers to be the functional units of the neo-cortex. Implicit in re-scaling operations though is a loss of degrees of freedom. However, in reducing the dimensionality of the EP to a comparable order to that of the molar phenomena of behavior, it is argued that this loss may be regarded constructively as potentially facilitating the successful forging of mind-brain correlations. Various techniques are reviewed for extracting the dimensions of scalp activity, including those based on electric field theory which are argued to have high face validity. The multivariate approach is also surveyed. Principal components analysis, in particular, is evaluated using simulated data. The limitations of the method are pointed up. Caution is advised in the design of a PCA, and in the interpretation of factors. The use of discriminant analysis in combination with MANOVA is suggested as a more appropriate multivariate strategy. PMID- 6282349 TI - Prolactin in the marsupial Macropus eugenii, during the estrous cycle, pregnancy and lactation. AB - An heterologous double antibody radioimmunoassay (RIA) using a guinea-pig antiserum (33-9) raised against human prolactin and 125I-ovine prolactin has been developed to measure prolactin (Prl) in plasma and pituitary preparations of marsupials. In this system, purified tammar and kangaroo Prl preparations showed parallel dose-response curves as did serial dilutions of crude pituitary homogenates of tammar, possum and eastern grey kangaroo. Serial dilutions of plasma from ovariectomized and lactating female and castrate male tammars showed immunoreactivity, and plasma Prl levels increased after injection of TRH. The assay has been used to monitor changes in plasma Prl in female tammars in various reproductive states. Plasma Prl remained at basal concentrations of 20 to 30 ng/ml throughout the estrous cycle, at estrus and during pregnancy. However, just prior to parturition, there was a 2- to 3-fold increase in Prl concentrations which declined to basal levels after birth. During early lactation, Prl levels were low but increased to maximum concentration in the second half of lactation. PMID- 6282350 TI - Chromosomal analyses after in vitro fertilization of squirrel monkey (Saimiri sciureus) oocytes. AB - The effect of 1 microM dibutyril cAMP (cAMP media) on the chromosomal normality of in vitro fertilized squirrel monkey oocytes was examined. A total of 877 oocytes were collected laparoscopically 15-16 h after hCG injection from hormonally-induced follicles of 65 squirrel monkeys. Of these, 330 (37.6%) were prepared between 6 and 30 h after insemination and analyzed. Sperm penetration and the presence of a swollen sperm head in the ooplasm were observed by 6 h, and the nucleate stage by 10 h. Addition of 1 microM dbcAMP did not affect the oocyte maturation rate, however the in vitro fertilization rate was increased 26.5% over controls (46.3 vs. 36.6%). The first cleavage metaphase was found at 16 h after insemination. Between 24 and 30 h post-insemination, 76.6% of fertilized oocytes had reached the first cleavage metaphase. An incidence of triploidy (including dispermy) of 11.7% was observed. After chromosomal analysis of metaphase I, univalents were observed in 16.7% of the cases. Aneuploidy (8.9%) was also found in metaphase II. Of the first cleavage metaphases, 88.0% had the diploid number of chromosomes. Aneuploidy occurred 12.0% of the time and 5 cases of triploidies were observed. PMID- 6282351 TI - Inhibitory action of cyclic guanosine 5'-phosphoric acid (GMP) on oocyte maturation: dependence on an intact cumulus. AB - Oocyte cumulus complexes (OCC) were isolated from antral follicles of proestrous hamsters prior to the luteinizing hormone (LH)/follicle-stimulating hormone (FSH) surge and then incubated from 4 to 24 h at 37 degrees C. Under these conditions approximately 89% of the oocytes exhibited metaphase plates (11% were in the dictyate stage) and addition of LH (200 ng/ml ovine NIH-LH-S20) did not significantly alter the percentage of oocyte maturation. Approximately 50% of the OCC incubated for 24 h with 6 mM 8-bromo-cyclic quanine 3':5' monophosphate (8-Br cGMP) were prevented from maturing beyond the dictyate stage. OCC incubated with 6 mM 8-bromo-cyclic adenosine 3':5' monophosphate (8-Br-cAMP) were also prevented from maturing (59.5% 8-Br-cAMP) in vitro. LH (200 ng/ml) was able to overcome the 8-Br-cAMP-induced inhibition of oocyte maturation in OCC. It also produced a decrease in 8-Br-cGMP mediated inhibition which was more pronounced when the dosage of LH was increased to 10 microgram/ml. 8-Br-cGMP prevented oocyte maturation in a dose and time dependent manner. Only 8-Br-cAMP prevented maturation (approximately 60% inhibition) of denuded oocytes in vitro. Denuded oocytes incubated with and without 8-Br-cGMP exhibited no inhibited of maturation. These results indicate that 8-Br-cGMP may exert its inhibitory effects through the cumulus cells. On the other hand, cAMP appears to directly inhibit oocyte maturation. PMID- 6282352 TI - Pituitary and ovarian function in postpartum beef cows. I. Effect of suckling on serum and follicular fluid hormones and follicular gonadotropin receptors. AB - The effect of suckling on serum and follicular fluid hormones and on follicular gonadotropin receptors was studied. Sixteen anestrous postpartum cows were assigned to 1 of 2 groups: suckled (S) or weaned (W). All calves were allowed to suckle ad libitum from parturition to 21 days postpartum when calves from W cows were weaned. All cows were ovariectomized on Day 25 postpartum. W cows had more (P less than 0.01) pulses of LH during the 96-h period from weaning until ovariectomy than S cows (6.3 vs. 1.3 pulses). Serum concentrations of prolactin (Prl), estrone (E1), estradiol-17 beta (E2) and progesterone (P) were not different (P greater than 0.10) between groups. Furthermore, there were n differences (P greater than 0.10) in follicular in contents of luteinizing hormone (LH), E1, E2 and P between the treatment groups. However, follicular fluid content of Prl was greater (P less than 0.05) in the W cows than in the S cows (123 vs. 65.1 ng/cow). The number of follicular LH receptors was greater (P less than 0.05) in the W cows than in the S cows (71.1 vs. 48.3 fmoles/mg protein) although the number of follicular follicle-stimulating hormone (FSH) receptors was not different (P greater than 0.10) between W cows and S cows (1531 vs. 1862 fmoles/mg protein). There were no correlation between serum hormone concentrations and follicular fluid hormone content; however, the numbers of follicular LH receptors and follicular fluid Prl content were highly correlated in the W cows (r = 0.85; P less than 0.05). It is concluded that removal of the suckling stimulus increases pulsatile LH release and the accumulation of Prl in the follicular fluid. These factors, either together or separately, may at least in part be responsible for the increase in follicular LH receptor concentrations that were observed in the W cows. PMID- 6282353 TI - Pituitary and ovarian function in postpartum beef cows. II. Endocrine changes prior to ovulation in suckled and nonsuckled postpartum cows compared to cycling cows. PMID- 6282354 TI - Regulation of protein phosphorylation and motility of sperm by cyclic adenosine monophosphate and calcium. AB - Motility and protein phosphorylation have been measured under identical experimental conditions in ejaculated dog sperm lysed with low concentrations of Triton X-100 and reactivated with [gamma-32P]ATP. Cyclic AMP stimulates motility and protein phosphorylation while calcium inhibits motility and the overall incorporation of phosphate into endogenous proteins. Analysis of 32P-labeled sperm proteins on 1- and 2-dimensional polyacrylamide gels demonstrates that an enhanced phosphorylation of a defined number of specific proteins is associated with cAMP-stimulated motility. A major axonemal proteins, namely tubulin, has been tentatively identified as a phosphoprotein subject to regulation by cAMP. The phosphorylation of tubulin is almost completely dependent upon cAMP and is not affected by microM calcium. On the other hand, the cAMP-dependent stimulated phosphorylation of the other sperm proteins still occurs, but in most instances at a reduced rate in the presence of calcium. Two high molecular weight (Mr) phosphoproteins (350,000 and 260,000 daltons) whose phosphorylation states are modified by cAMP and calcium also were identified. It is suggested that 1 or both these proteins may be high Mr subunits of dynein. The phosphorylation of 1 of these proteins is stimulated by cAMP, but not affected by calcium; the other is stimulated by cAMP and inhibited by calcium. Three major cAMP-independent phosphoproteins of Mr 98,000, 43,000 and 26,000 have been identified. The phosphorylation of the 98,000 Mr protein is markedly reduced by micromolar calcium and not restored by cAMP. Using anticalmodulin drugs to inhibit motility, we suggest that the inhibitory effects of calcium on flagellar motility may be mediated in part by calmodulin. We conclude that the regulation of flagellar motility in cAMP and calcium includes mechanisms involving the control of the phosphorylation state of sperm proteins, some of which may be axonemal components. PMID- 6282355 TI - Alcohol-induced Cushingoid syndrome. AB - Although it has been known for some time that alcohol affects the hypothalamic pituitary-adrenal axis, the clinical significance of this disturbance has only recently become recognised. Over the last 5 years, 27 cases of an apparent Cushing's syndrome have been documented, usually resolving within weeks of alcohol withdrawal. Differentiation from the true Cushing's syndrome can be difficult before resolution, which may be prolonged for several months, but an insulin stress test can be useful in their distinction. The syndrome is invariably pituitary dependent but may not be a specific drug effect of ethanol but rather a consequence of the chronic stress of alcoholism. PMID- 6282356 TI - Interactions of DNA with divalent metal ions. IV. Competitive studies of Mn2+ binding to AT- and GC-rich DNAs. PMID- 6282358 TI - Involvement of selection and amplification mechanisms in hormone receptor development in a unicellular model system. AB - It was demonstrated earlier, that long lasting exposure of Tetrahymena to a hormone (histamine) resulted in an increased responsiveness to a later re exposure. However, it was difficult to establish whether selection or amplification plays a role in receptor differentiation. As diiodotyrosine (T2) enhances the growth of Tetrahymena, in the present experiment the effect of T2 treatment on a long-term culture of Tetrahymena pyriformis was analysed by mathematical-statistical methods to differentiate the effects of selection and amplification mechanisms on hormone receptor development. Although continuous and periodic treatment with T2 enhanced cell division equally, the resulting populations differed in structure. On continuous treatment the population tended to become inhomogenous. The variance tended to increase for 9 days and decreased afterwards without, however, returning to the control level. On periodic treatment the variance was the same as in the control group, but the second and third exposure were significantly more effective than the first treatment, suggesting that the primary encounter with the hormone had given rise to lasting alterations (hormonal imprinting). It follows that continuous exposure involves a selection process which does not, however, account for a steady increase of the growth rate; for initial amplification, taking place also in this condition, and selection which takes effect later, compensate one another's effects. Regarding the unicellular experimental system as a phylo- and ontogenetic model, the conclusion lies close at hand that the selection and amplication mechanisms promote hormone receptor development by joint rather than alternate action. PMID- 6282357 TI - Solute transport at the plasmalemmma and the early evolution of cells. PMID- 6282359 TI - [Role of cyclic adenosine 3,5-monophosphate and prostaglandin E2 in the mechanism of the inhibitory action of nicotinic acid on thrombocyte aggregation]. PMID- 6282360 TI - [Determination of myelin marker enzymes in the blood serum in peripheral nervous system diseases]. AB - The activity of 2',3'-cyclonucleotide-3-phosphohydrolase (CNP) and leucine aminopeptidase (LAP) was studied for the first time in the blood of patients with diseases of the peripheral nervous system. A spectrophotometric method for CNP activity detection was described. The activity absent from the blood of normal subjects was detectable with statistical significance in the patients with the diseases in question. Analogous results were obtained in the animals with an experimental injury to the myelin coat of the peripheral nerves. Study into the activity of LAP in the pathological conditions under consideration revealed no significant deviations. It is emphasized that CNP may transfer to the blood serum from myelin decay foci and that the method described may be used for diagnostic purposes. PMID- 6282361 TI - [Effect of thymalin on the cyclic nucleotide system in the mouse spleen]. AB - The effect of the thymus preparation thymalin (1000 micrograms/mouse, i.p.) on the system of cyclic nucleotides in the spleen has been studied. The preparation produced two-phasic changes in the content of cAMP and cGMP, as well as in the activity of cAMP phosphodiesterase. The changes in the spleen cAMP content are consequent on those in the activity of phosphodiesterase. PMID- 6282363 TI - Phagocytosis-induced chemotaxis receptor cycling in neutrophils is mediated by thiol oxidation. AB - We previously demonstrated that neutrophil (PMN) phagocytosis of opsonized zymosan (OPZ) caused oxygen-dependent inhibition of chemotactic peptide receptor (CPR) binding using the ligand 3H-formyl-methionyl-leucyl-phenylalanine (3H FMLP). In the current studies we sought to determine the mechanism of CPR inhibition by OPZ. We found that 3 mM cysteine and 5 mM dithiothreitol (DTT) did not decrease PMN phagocytosis, but abolished CPR inhibition by OPZ. Furthermore, incubation of PMN with DTT after OPZ partially restored the suppressed CPR. PMN CPR saturation studies with 3H-FMLP on PMN incubated with or without DTT after phagocytosis indicated that DTT restored receptor numbers to 92% +/- 6% of prephagocytosis values and also improved CPR affinity for 3H-FMLP. In additional studies we found that the cell penetrating thiol reagents mercuric chloride (HgCl2), N-ethylmaleimide (NEM), and diamide, but not the nonpenetrating agents p chloromercuriphenylsulfonic acid and p-chloromercuribenzoate, caused rapid, dose dependent, DTT-inhibitable suppression of up to 85% CPR binding. CPR inhibition by HgCl2 and NEM was irreversible. PMN CPR saturation studies showed that NEM decreased total receptor numbers, not CPR affinity. The effect of NEM was not inhibited by receptor occupancy at 4 degree C or at 37 degree C. These studies indicate that: (1) intact thiol groups are a requirement for CPR ligand-binding; (2) the thiol groups are presumptively located on the interior side of, or within hydrophobic portions of, the plasma membrane and are not part of the 3H-FMLP binding site; (3) thiol oxidation-reduction induces reversible alteration in CPR binding; (4) phagocytosis-induced modulation of the CPR is due, in part, to thiol oxidation. These studies suggest that postphagocytic, thiol-mediated reduction in CPR binding may play a physiologic role in the inhibition of PMN chemotaxis at inflammatory sites. The state of CPR-associated thiols may be an important determinant of CPR binding under physiologic conditions other than phagocytosis. PMID- 6282362 TI - Metabolic, membrane, and functional responses of human polymorphonuclear leukocytes to platelet-activating factor. AB - The phospholipid mediator of anaphylaxis, platelet-activating factor (PAF) is chemotactic for polymorphonuclear leukocytes (PMN). We have examined this agent's effects on several other PMN functions. Human PMN were prepared from heparinized venous blood by Ficoll gradient. Metabolic burst was examined by measurement of O2 use and O2.- production in the presence or absence of PAF (10(-6)--10(-9) M). Unless cells were treated with cytochalasin-B (5 micrograms/ml), no significant respiratory burst was demonstrated. However, pretreatment with PAF (10(-7) M) enhanced approximately threefold the O2 utilization found when cells were subsequently stimulated with 10(-7) M FMLP. PAF also stimulated arachidonic acid metabolism in 14C-arachidonic acid-labeled PMN. Thin-layer chromatography analysis of chloroform-methanol extracts showed substances that comigrated with authentic 5-hydroxyeicosatetraenoic acid had a marked increase in radioactivity following PAF stimulation at 10(-7) M. PAF failed to stimulate release of granule enzymes, B-glucuronidase, lysozyme, or myeloperoxidase unless cytochalasin-B were added. PAF from 10(-6) M to 10(-10) M affected PMN surface responses. PMN labeled with the fluorescent dye, chlorotetracycline, showed decreased fluorescence upon addition of PAF, suggesting translocation of membrane-bound cations. Further, the rate of migration of PMN in an electric field was decreased following PAF exposure, a change consistent with reduced cell surface charge. PMN self aggregation and adherence to endothelial cells were both influenced by PAF (10( 6) M--10(-9) M). Aggregation was markedly stimulated by the compound, and the percent PMN adhering to endothelial cell monolayers increased almost twofold in the presence of 10(-8) M PAF. Thus, PAF promotes a variety of PMN responses: enhances respiratory burst, stimulates arachidonic acid turnover, alters cell membrane cation content and surface charge, and promotes PMN self-aggregation as well as adherence to endothelial cells. PMID- 6282364 TI - A monoclonal immunoblastic sarcoma in donor cells bearing Epstein-Barr virus genomes following allogeneic marrow grafting for acute lymphoblastic leukemia. AB - A patient undergoing marrow grafting for acute lymphoblastic leukemia from his partially HLA-mismatched sister displayed a widely disseminated immunoblastic sarcoma at autopsy. The tumor was monoclonal by immunoglobulin light-chain staining. Blot hybridization analysis, using a cloned highly polymorphic locus in human DNA as a probe, showed the tumor to be of donor-cell origin. Cytogenetic analysis also demonstrated donor-cell origin. Blot hybridization analysis demonstrated Epstein-Barr virus (EBV) genomes in the tumor. By contrast, reexamination of material from a previously reported case of a donor-type relapse showed no evidence of EBV DNA. In neither case was there evidence of cytomegalovirus DNA. This study documents the association of EBV with a malignant, monoclonal B-cell lymphoma arising in a marrow graft recipient. We conclude that DNA restriction fragment length polymorphisms can be used to prove the origin (donor or host) of neoplastic relapse following allogeneic marrow grafting. Further, cell types different from those of the original leukemia may be involved. PMID- 6282365 TI - Induction of human platelet fibrinogen receptors by epinephrine in the absence of released ADP. AB - The ability of epinephrine to expose platelet fibrinogen receptors independent of released ADP was assessed using aspirin-treated, gel-filtered platelets. Similar to ADP-induced aggregation, platelet aggregation in response to epinephrine was accompanied by fibrinogen binding. Ten micromolar epinephrine induced a maximum number of platelet fibrinogen receptors in the absence of significant 14C serotonin release. As indicated by Scatchard analysis, receptors exposed by both epinephrine and ADP had similar affinities for fibrinogen, but epinephrine induced approximately 30% fewer receptors than did ADP. This appears to correlate with the lesser degree of primary aggregation observed with this agent. Studies using phentolamine, a specific alpha-adrenergic antagonist, apyrase, or creatine phosphate/creatine kinase indicate that the exposure of platelet fibrinogen receptors by epinephrine was specific for platelet alpha-adrenergic receptor stimulation and was not the result of released ADP. PMID- 6282367 TI - Why inclusion bodies do assume different locations in thalassaemic erythrocytes. AB - The reported scanning (SEM), transmission (TS), and freeze-etching (FE) electron microscopic studies have agreed in confirming that in thalassaemic erythrocytes, previously incubated with brilliant cresyl blue (BCB), the unpaired alpha chains precipitate in the central portions of the cell whereas excess beta chains locate in the submembranous regions. This is due to the fact that beta chains, possessing two thiols instead of only one (as in alpha chains), are more liable to bind to similar groups contained in the inner red cell leaflet. Less soluble alpha chains tend to form inter-chain bridges and thus precipitate centrally. SEM observations have given evidence that on the surface of the affected red cells denaturated alpha chains give rise to large and shallow invaginations whereas denatured beta chains lead to diffuse wrinkled appearance. The causes of the different SEM aspects have been suggested. PMID- 6282368 TI - An outbreak of epidemic keratoconjunctivitis in Antwerp, caused by an orbiviruslike particle. PMID- 6282366 TI - Angiotensin II-induced relaxation of vascular smooth muscle. AB - The effects of angiotensin II (AII) on contractile tension were studied in vascular smooth muscle from dogs, pigs and rabbits. Helically cut strips of renal veins were mounted in organ chambers and isometric contractions were recorded. Contraction of the venous strips was induced by application of 10(-8) g/ml norepinephrine (NE). Subsequent addition of 5 X 10(-8) g/ml AII caused a triphasic response: (1) there was an initial contraction (subsequent contractions were tachyphylactic in all species); (2) the contraction was followed by a relaxation below the contraction induced by NE (subsequent relaxation responses were tachyphylactic in dog and pig veins), and (3) there was a return from the relaxation to the level of the NE-induced contraction. The duration of the entire response was approximately 5 min. The magnitude of the relaxation varied inversely with the level of the NE contraction when the contractile state was altered by changing the NE concentration. Conditions which inhibit the sodium pump (potassium-free solution and ouabain) and beta-adrenergic blockade with propranolol had no effect on the AII-induced relaxation. The relaxation was temperature sensitive. Inhibitors of prostaglandin synthesis (indomethacin and aspirin) and saralasin attenuated the relaxation in response to AII. Prostaglandins E1 and E2 and arachidonic acid caused relaxation of renal vein strips contracted with NE; the relaxant effect of arachidonic acid was blocked by indomethacin. These results suggest that: (1) All stimulates the synthesis of prostaglandins in isolated venous smooth muscle, and (2) endogenous prostaglandins modulate the response of venous smooth muscle to AII. PMID- 6282369 TI - [Aniridia and Wilm's tumor]. PMID- 6282370 TI - Familial dermal cylindroma with involvement of the parotid gland. AB - The association of turban tumour with other dermal cylindromas in the parotid gland is very rare. Another case is reported in which the hereditary nature of the disease is discussed and certain aspects of the treatment and management of these cases are considered. PMID- 6282371 TI - Alteration in the level of endogenous hypothalamic prostaglandins induced by delta 9-tetrahydrocannabinol in the rat. AB - 1 Whole brain and regional brain levels of prostaglandin E2 (PGE2)-like material have been determined following administration of delta 9-tetrahydrocannabinol (delta 9 -THC) in rats. 2 Intravenous administration of delta 9-THC 2 mg/kg, resulted in marked behavioural changes and hypothermia. The behavioural changes consisted mainly of catatonia (most apparent at 30 min after administration of delta 9-THC), followed by sedation (most evident at 60 min). Hypothermia was marked from 30 min after administration of delta 9-THC. 3 delta 9-THC did not after the whole brain levels of PGE2-like material 30, 60 or 120 min after administration. 4 delta 9-THC did not alter the levels of PGE2-like material in the medulla oblongata/pons, midbrain, cortex and cerebellum, 30 min after administration. However, there was a significant reduction of PGE2-like material in the hypothalamus, 30 min after delta 9-THC. 5 It is suggested that the delta 9 THC-induced decrease in hypothalamic PGE2-like material may contribute to the hypothermia observed following delta 9-THC administration. PMID- 6282372 TI - Relation of plasma renin activity to the antihypertensive effect of MK 421 in the rat. AB - 1 The effect of the angiotensin converting enzyme inhibitor, MK 421 (N-((S)-1 (ethoxycarbonyl)-3-phenylpropyl)L-Ala-L-Pro), on the blood pressure of two-kidney Goldblatt hypertensive rats has been investigated in relation to he initial plasma renin activity (PRA) and the initial blood pressure of the individual animals. 2 Blood pressure was monitored by an indirect tail-cuff method at 1, 3, 6 and 24 h after dosing. MK 421 produced a fall in blood pressure in the majority of animals, but the extent of this reduction varied considerably between individuals. 3 The change in blood pressure showed a significant correlation with both the initial PRA and the initial blood pressures of the animals. However, only a modest correlation was found between the initial PRA and the degree of hypertension. 4 MK 421 (10 mg/kg, orally) produced a mean blood pressure change which was statistically significant (P less than 0.001) at all times tested. 5 It is concluded that the degree of antihypertensive activity of MK 421 is related to the degree of activity of the renin-angiotensin system which, in this model at least, is reflected by the PRA. PMID- 6282373 TI - Glucocorticoids induce the formation and release of anti-inflammatory and anti phospholipase proteins into the peritoneal cavity of the rat. AB - 1 Dexamethasone and hydrocortisone induced the release of anti-phospholipase proteins into the peritoneal cavities of rats. 2 Adrenocorticotrophic hormone (ACTH) also releases these proteins in normal but not in adrenalectomized rats. 3 Peritoneal lavage proteins were separated by ion-exchange and size exclusion chromatography. The anti-phospholipase activity occurred in four separate fractions with the major component having an apparent mol. wt. of 40 k. 4 Column fractions containing these anti-phospholipase proteins had anti-inflammatory effects in the rat carrageenin pleurisy model whereas other fractions were inactive. 5 The proteins appear to be identical to macrocortin and lipomodulin, the 'second messengers' of glucocorticoid hormone action on the arachidonate system. PMID- 6282374 TI - Generation of leukotriene B4, its all trans isomers and 5-hydroxyeicosatetraenoic acid by rat basophilic leukaemia cells. AB - 1 When rat basophilic leukaemia (RBL-1) cells were incubated for 4 min, in the presence of arachidonic acid (AA) and calcium ionophore A23187, three isomers of leukotriene (LT) B4 and 5-hydroxyeicosatetraenoic acid (5-HETE) were generated. 2 The isomers of LTB4 separated by reverse phase high pressure liquid chromatography (h.p.l.c.) were found to be 5 (s)-dihydroxy-6, 14-trans-8,10-trans eicosatetraenoic acid: 5(S), 12(R)-dihydroxy-6,14-trans-8,10-trans eicosatetraenoic acid and 5(S),12(R)-dihydroxy-6,14-cis-8,10-trans eicosatetraenoic acid or LTB4. 3 5(S), 12(R)-cis, trans, trans-LTB4 produced by RBL-1 cells caused aggregation of rat polymorphonuclear leucocytes (PMNs). Its other two isomers (5(S), 12(S)-all trans-LTB4; 5(S), 12-(R)-all trans-LTB4) were much less active and 5-HETE was found to be inactive. 4 5(S), 12(R)-cis, trans, trans-LTB4 formed from RBL-1 cells contracted guinea-pig parenchymal strips (GPPs) and was indistinguishable from synthetic 5(S), 12(R)-cis, trans, trans LTB4. The other two isomers of LTB4. i.e. 5(S), 12(S)-all trans-LTB4 and 5(S), 12(R)-all trans-LTB4 as well as 5-HETE were inactive on GPP. 5 RBL-1 LTB4 was equipotent with LTD4. in most experiments, in contracting GPP but had no activity on guinea-pig ileum smooth muscle (GPISM). 6 RBL-1 cells contain the enzyme systems necessary for generating LTB4 as well as the leukotriene(s) with amino acid residues at C-6. PMID- 6282376 TI - Can drugs help patients with lower limb ischaemia? AB - The prevalence of symptomatic arterial disease of the lower limbs is 2 per cent of the population aged 45-60, but it has a relatively benign course, with 70 per cent of patients requiring no therapy. Of the numerous drugs used in the treatment of the disease, there is no evidence to suggest that antilipaemic drugs, anticoagulants, vasodilators or rheological agents confer any benefit to the patient. The initial use of antiplatelet drugs and prostaglandins has been disappointing despite the undoubted importance of the platelet/endothelial interaction in the aetiology of atherosclerosis. As it is unlikely that we can reverse advanced disease, this is hardly surprising. Long term use of these drugs may prevent deterioration in those patients with progressive disease, and controlled trials on this aspect of treatment are now required. Symptomatic relief in the claudicant may perhaps be obtained with naftidrofuryl and suloctodil and with the former in more severe ischaemia, but their use should not replace the beneficial effects of exercise and cessation of smoking. PMID- 6282377 TI - Adverse reactions to nitrofurantoin in the United Kingdom, Sweden, and Holland. AB - A study was carried out comparing the reporting of adverse reactions to nitrofurantoin in Sweden, Holland, and the United Kingdom. The Swedish drug regulatory authorities receive more reports of adverse reactions to nitrofurantoin than to any other drug, and the annual reporting rate is increasing despite a steady fall in the prescription of nitrofurantoin. In the United Kingdom the total number of adverse reaction to nitrofurantoin reported to the Committee on Safety of Medicines has been steadily falling for many years, as has the number of prescription. Lung reactions and liver damage are more commonly reported in Sweden than in the UK, where gastrointestinal symptoms and peripheral neuropathy are more common. Such figures as were available in Holland showed an intermediate picture. The incidences of blood dyscrasias and allergic reactions are similar in the three countries. The causes for such differences are unresolved, but it is disturbing that the incidence and range of adverse reactions to such a common drug as nitrofurantoin reported to the national authorities can differ so much in countries that have close ethnic, dietary, and economic similarities. Thus great caution must be observed when data on adverse drug reactions from the country are extrapolated to another. PMID- 6282378 TI - Hypophosphataemic vitamin-D resistant rickets may need phosphate supplements. PMID- 6282375 TI - Adenosine 5-diphosphate antagonists and human platelets: no evidence that aggregation and inhibition of stimulated adenylate cyclase are mediated by different receptors. AB - 1 Adenosine 5'-diphosphate (ADP) induces human platelet aggregation and noncompetitively inhibits stimulated human platelet adenylate cyclase; it has been suggested that these two effects are mediated by separate ADP receptors on the platelet surface. 2 Adenosine 5'-triphosphate and seven adenine nucleotide analogues were tested as inhibitors of both effects of ADP on human platelets, and were found to be competitive. 3 pA2 values were calculated for each antagonist for inhibition of both effects of ADP, and a good correlation (correlation coefficient 0.87; p less than 0.01) was found between the pA2 values for inhibition of ADP-induced aggregation and the pA2 values for inhibition of the effect of ADP on stimulated adenylate cyclase. 4 Such a correlation does not support the suggestion that ADP-induced aggregation and the inhibition by ADP of stimulated adenylate cyclase are mediated by two separate receptors. PMID- 6282379 TI - Prophylactic cephalosporin in prostatic surgery. PMID- 6282380 TI - Of woodchucks and men: the continuing story of hepatitis B and hepatocellular carcinoma. PMID- 6282381 TI - Effect of high-fibre diet on haemostatic variables in diabetes. PMID- 6282382 TI - "Scalded mouth" caused by angiotensin-converting-enzyme inhibitors. PMID- 6282383 TI - Prenatal serological diagnosis of intrauterine cytomegalovirus infection. PMID- 6282385 TI - Specificity of the immunosuppressive action of carbimazole in Graves's disease. PMID- 6282384 TI - Selective infection of lower respiratory tract by respiratory viruses in children with recurrent respiratory tract infections. AB - Thirty preschool children presenting with recurrent respiratory infections and their unaffected siblings were observed prospectively for a year. The index children experienced more episodes of acute respiratory infection than their siblings. Respiratory viruses were the major cause of respiratory infections. The index children had lower respiratory tract disease, predominantly wheeze, during 34% of proved respiratory virus infections compared with 11% of such infections experienced by the control children (p less than 0.02). Atopic children had an increased tendency to wheeze that did not reach significance, but atopy was not associated with increased susceptibility to respiratory infections. PMID- 6282386 TI - Primary biliary cirrhosis. PMID- 6282387 TI - Clinical observations on bovine papillomatosis (warts). PMID- 6282388 TI - Changes produced by a hypomyelinating neuropathy in muscle and its innervation. Morphological and physiological studies in the Trembler mouse. AB - The peripheral nerves of the Trembler mouse are hypomyelinated. The effects of the neuropathy on muscle and motor end-plate morphology, and on neuromuscular transmission in slow-twitch (soleus) and fast-twitch (extensor digitorum longus) muscle are reported. After forming normally, motor end-plates developed ultraterminal sprouts by about 18 days of age in deep, slow-twitch muscles. The only ultrastructural abnormality in muscle at this age was disruption of Z-lines. Ultraterminal sprouting progressed, and by about 3 months the innervation zone consisted of a mass of branching axons associated in places with cholinesterase activity. The ultrastructure of end-plates became abnormal. Fascicular atrophy was present in older Tremblers and the ultrastructural morphology of muscle fibres became disorganized with accumulation of subsarcolemmal granular material. Abnormal muscle fibres were innervated by axonal sprouts. The superficial, predominantly fast-twitch, muscles showed milder changes at all ages even though the nerves supplying them were hypomyelinated. Studies of neuromuscular transmission showed that soleus retained its slow-twitch and extensor digitorum longus its fast-twitch characteristics, and miniature end-plate potentials were of normal frequency and amplitude. The latency of end-plate potentials and the refractory period of transmission were prolonged in both soleus and extensor digitorum longus. With repetitive stimulation at 50 Hz a cyclical pattern of responses and failures occurred which was probably caused by intermittent conduction block along the peripheral nerve. Although the pathological changes in Trembler muscle were like those of partial denervation, atrophic muscles contained an abundance of axons and there was no evidence of axonal degeneration. The changes in muscle are therefore a consequence of hypomyelination without axonal loss. Since slow-twitch muscles are normally subjected to prolonged stimulation, failure of Trembler axons to conduct sustained trains of stimuli in vivo may contribute to the development of pathological changes in slow-twitch muscle. Hypomyelimated axons may be capable of conducting short bursts of impulses, however, which is the pattern of stimulation in fast-twitch muscle in vivo, so that fast-twitch muscle fibres and end-plates remain relatively spared. PMID- 6282389 TI - Brain adrenergic receptors and resistance to stress. AB - The relationship between brain beta-adrenergic receptors and adaptation to stress was studied in rats subjected to repeated restraint stress. The stress was found to produce a reduction in the density of these receptors in the hypothalamus, cerebral cortex and brain stem. This change first appeared after 4-7 days and persisted for the duration of the two-week stress. Adaptation, as judged by resistance to the anorexic and gastric lesion-inducing effects of the stress, occurred progressively over the full two-week period. The loss of beta-receptors was found to correlate positively with the degree of adaptation. The relationship was strongest for the hypothalamus but was also apparent in the cortex and brain stem. These findings support the hypothesis that a reduction in the number of brain adrenergic receptors is one of the biochemical factors underlying adaptation to stress. PMID- 6282391 TI - Intrahypothalamic colchicine infusions disrupt lordotic responsiveness in estrogen-treated female rats. AB - Since some estrogenic effects on lordotic responsiveness are mediated through hypothalamic protein synthesis, we conducted experiments to determine if axoplasmic transport in the hypothalamus is necessary for the induction and maintenance of this reflex by estrogen. Colchicine infusion into the hypothalamus, but not into the dorsal thalamus, of ovariectomized rats 24 h prior to administration of subcutaneous estrogen implants delayed the induction of lordotic responsiveness, as measured by the manual (cutaneous-pressure) method, by 2 days, as compared with vehicle-infused rats. In other experiments, colchicine infusion into the hypothalamus, but not into the dorsal thalamus, of conscious, ovariectomized, estrogen-implanted rats displaying maximal lordotic responsiveness resulted in a bimodal decline in lordotic responsiveness. An initial decline occurred 20-40 min after infusion, and was associated with general behavioral agitation and hyperactivity. A subsequent decline began 4 h after infusion and lasted for several days. Vehicle infusion did not decrease lordotic responsiveness. Colchicine infusion did not alter multiunit electrical activity recorded near hypothalamically directed cannulae tips over a period of several hours. Results suggest that axoplasmic transport within and/or from the hypothalamus is necessary for the estrogenic induction and maintenance of the lordosis reflex in rats. PMID- 6282390 TI - An early sodium and a late calcium phase in the afterdischarge of peptide secreting neurons of Aplysia. AB - Following brief electrical stimulation of a pleuroabdominal connective nerve, the clusters of peptidergic bag cell neurons of Aplysia generate a long-lasting (or approximately 30 min) synchronous afterdischarge. In a normal seawater medium, the afterdischarge comprises an early phase of rapid firing (2-6 Hz) lasting for less than one minute, followed by a second, prolonged phase of lowering firing rate (less than 0.5 Hz). We have found that the transition from the first, rapid firing phase to the prolonged second phase of afterdischarge is associated with an increase both in the width and height of intracellularly recorded action potentials. In addition, we find that in the presence of the potassium channel blocker, TEA, such biphasic afterdischarges may be triggered by depolarizing current that evokes action potentials in a single bag cell neuron in a cluster. Afterdischarges resembling either the first or the second phase of a normal afterdischarge may be produced by using media deficient in calcium or sodium, respectively, in combination with potassium channel-blockers. Brief stimulation in media deficient in calcium give rise to afterdischarges of high firing frequency (2-6 Hz) lasting for less than one minute (mean duration = 41 s) while, in a sodium-free medium, longer afterdischarges (mean duration = 18 min) of lower firing rate (less than 0.3 Hz) were generated. At the end of afterdischarge in a normal seawater medium, containing both sodium and calcium ions, the bag cell neurons become refractory to stimulation. Thus, further stimulation either fails to induce afterdischarge or results in afterdischarges that are attenuated both in frequency of firing and in duration compared with the first afterdischarge. In these experiments we found that the brief afterdischarges, observed in low calcium media, which resemble the first phase of a normal afterdischarge, did not induce subsequent refractoriness while afterdischarges evoked in the normal calcium, sodium-free medium did result in refractoriness to further afterdischarge. Our data indicate that both phases of a bag cell afterdischarge are due to intrinsic bag cell mechanisms and suggest that the firing pattern in the first phase is largely sodium dependent and that of the second, slow phase is calcium dependent. Moreover, our data indicate that it is the second, calcium dependent phase that induces the refractoriness that follows bag cell afterdischarge. PMID- 6282392 TI - Intra-axonal recordings in rat dorsal column axons: membrane hyperpolarization and decreased excitability precede the primary afferent depolarization. AB - Intra-axonal recordings were obtained in the dorsal columns of the rat lumbosacral spinal cord. Dorsal root or dorsal column stimulation at levels subthreshold for the impaled axon elicited a prolonged depolarization corresponding to the primary afferent depolarization (PAD). The depolarization was preceded by a brief hyperpolarizing potential during which excitability was decreased. The hyperpolarization corresponds temporally to the extracellularly recorded DRP IV component of the dorsal root potential described by Lloyd and McIntyre, and may represent the intracellular correlate of this potential. Possible mechanisms for this hyperpolarization include electrical interactions between neuronal elements, a biphasic GABA response, or attenuation of background afferent axonal depolarization. PMID- 6282393 TI - Dye-coupling between pyramidal cells of rat hippocampus in vivo. AB - Lucifer Yellow was intracellularly injected into pyramidal cells of rat hippocampus in vivo. In 10 successful staining experiments there were 5 cases of dye-coupling, in which 2-4 cells were stained though only one was injected. These results indicate that dye-coupling and, therefore, presumably electrotonic coupling, occurs between neurons of the hippocampus in the whole animal and confirms previous findings in the hippocampal slice in vitro. PMID- 6282395 TI - Physiological correlates of multiple subtypes of enteric opiate receptor; functional analysis of myenteric delta receptors. PMID- 6282394 TI - On the use of lesions of afferents to localize neurotransmitter receptor sites in the striatum. AB - Rats received lesions of either the cortico-striatal or the nigrostriatal projections. Two to three days after either type of lesion, evidence for degenerating dendritic spines that were postsynaptic to degenerating striatal boutons was often encountered. This type of transsynaptic degeneration of spines indicates that it is inappropriate to conclude that decreases in the number of neurotransmitter receptors after these types of lesions necessarily indicates that these receptors are located presynaptically on striatal afferents. PMID- 6282396 TI - Aspartate-induced changes in extracellular free calcium in 'in vitro' hippocampal slices of rats. AB - Extracellular free calcium ([Ca2+]0) and neuronal activity were measured with double-barreled Ca2+-selective/reference microelectrodes in the CA1 field of rat hippocampal slices during stimulation of Schaffer collaterals and during iontophoretic application of the excitatory amino acid aspartate (Asp). Baseline [Ca2+]0 was 1.5 mM. Repetitive stimulation and aspartate application caused decreases in [Ca2+]0 (delta Ca), which were largest in the pyramidal cell layer. During iontophoretic application of GABA, aspartate often failed to induce significant alterations in [Ca2+]0. Mg2+ (9 mM) and Ni2+ (3mM) when added to the perfusion medium reduced aspartate-dependent delta Ca by 15 and 70% respectively. In tetrodotoxin (10(-6) M) containing Ringer the aspartate-induced delta Ca were about 10% smaller. These findings suggest that the decreases in [Ca2+]0 are mainly due to a postsynaptic Ca2+ entry through selective Ca2+ conductances. PMID- 6282397 TI - beta-Carboline inhibition of benzodiazepine receptor binding in vivo. AB - The in vivo potencies of 4 beta-carboline derivatives as inhibitors of benzodiazepine receptor binding were investigated. For all 4 derivatives maximal inhibition was seen within 2 min after i.v. application but decreased continuously for the next 20 min and after this time accounted for less than one third of maximal inhibition. The in vivo potencies of the beta-carbolines as inhibitors of benzodiazepine receptor binding are much smaller than one would expect from their affinities measured in vitro. Thus, it is assumed than only a small portion of the i.v. dose reaches the brain. PMID- 6282398 TI - Primary afferent units from the hairy skin of the rat hind limb. AB - The properties of cutaneous units in the saphenous nerve of the rat have been surveyed. We studied 137 units with myelinated (A-fibre) axons conducting at 4-44 m/s. Of the A-fibre units 66% gave a rapidly adapting discharge following hair movement and could be classified into categories similar to those described previously in cat and rabbit. Two categories predominated: (1) D-hair units with slowly conducting axons and relatively large receptive fields that responded to slow movement of hairs, including down hairs when these were present and (2) G hair units with larger axons and relatively small fields that were only excited by fast movement of guard hairs. Of the A-fibre units 20% were high threshold mechanoreceptors. As in other species, these had a wide range of conduction velocities and multi-point receptive fields. Other types of A-fibre units found were (a) sensitive, RA units not excited by hair movement, (b) relatively insensitive RA units with diffuse receptive fields and (c) slowly adapting mechanoreceptor units. We studied 149 units with unmyelinated (C-fibre) axons conducting at 0.49-0.89 m/s. Of the C-fibre units 73% were of the polymodal nociceptor type. They had small receptive fields and responded to pressure and heating. The average heat threshold was 47 degrees C (+/- 6 degrees C, S.D.). Units were often not sensitized by suprathreshold heating unlike similar units in cat and rabbit. Other C-fibre units found were sensitive mechanoreceptors (12%), cold thermoreceptors (4%) or were very insensitive or inexcitable (11%). The pattern of innervation of rat limb hairy skin resembles previously studied mammalian species. A notable feature is the large proportion of C-polymodal nociceptor units. In this respect the rat resembles the primate and differs from the cat. PMID- 6282399 TI - [Structural polypeptide VP1 in poliovirus type 1 induced by neutralizing antibodies]. AB - Anti-type 1 (Mahoney) poliovirus neutralizing antibodies were obtained by immunizing rabbits with structural polypeptide VP1. This polypeptide was isolated from the virion by PAGE-SDS electrophoresis and extracted from the gel by electroelution. Anti VP2 and anti VP3 sera prepared in the same was did not display any significant antipolio neutralizing activity. PMID- 6282400 TI - [Immunologic assessment and acoustic "stressing situation" in an experimental system BALB/c mice, the Tsc-3T3 tumor]. AB - Previous results in Hamsters showed enhancement of tumor takes in animals exposed to acoustic aggression. Since this phenomenon could be due to immunodepression, we investigated whether this was the case in a better defined system of inbred BALB/c Mice. The results show that in these conditions, the tumor graft could not transgress a minor histocompatibility antigen nor decrease the protective effects of polyoma virus against tumor challenge and that all the immune tests investigated were unaltered. PMID- 6282401 TI - [Determination of the terminal sequences of the transposon Tn7]. PMID- 6282403 TI - [Production of monoclonal antibodies against bovine rotavirus]. PMID- 6282402 TI - [Comparison of intracellular actin of thymosin alpha-1 and thymic serum factor]. PMID- 6282404 TI - [Further studies on the etiology of autumnal infantile gastroenteritis (author's transl)]. PMID- 6282406 TI - [Comprehensive therapy for small cell carcinoma on the lung---report of 18 cases (author's transl)]. PMID- 6282405 TI - [A pedigree of syndactyly and triphalangeal thumb in three consecutive generations (author's transl)]. PMID- 6282407 TI - Blood:bone equilibrium. PMID- 6282408 TI - Serum 1,25-dihydroxyvitamin D levels in subjects with X-linked hypophosphatemic rickets and osteomalacia. AB - In order to determine whether a defect in vitamin D metabolism might play a role in the pathogenesis of X-linked hypophosphatemic rickets and osteomalacia (XLH), we compared the serum 1,25-dihydroxyvitamin D [1,25(OH)2D] level in 52 normal subjects and 37 patients with XLH. In untreated patients, adults were found to have values similar to age-matched controls, while youths had values similar to growth-rate-matched controls but significantly lower than the levels of age matched controls who were growing at a normal rate. In contrast, treated XLH patients of all ages had serum levels significantly lower than both controls and untreated XLH patients. Further, the serum levels of 1,25(OH)2D in these treated patients had a significant inverse linear correlation with serum 25-(OH)D concentrations. We propose that subjects with XLH have serum 1,25(OH)2D levels within appropriate age- and growth-rate-matched normal ranges. However, in the presence of hypophosphatemia, we would have anticipated elevated levels of 1,25(OH)2D; viewed in this light the serum 1,25(OH)2D levels are inadequate, suggesting the presence of a relative deficiency of this active vitamin D metabolite. PMID- 6282409 TI - Blood: bone disequilibrium. V. Effects of a diphosphonate (EHDP) on phosphate fluxes in rat calvaria. AB - Calvaria taken from rats (normal or thyroparathyroidectomized) given a diphosphonate (EHDP 10 mg/kg daily for 7 days) were placed in special Ussing chambers for the measurement of phosphate fluxes to and from bone. When compared with appropriate controls, the experimental calvaria showed a marked decrease in influx, K, a significant increase in the projected equilibrium concentration, E/K, where E is the efflux and a significant increase in the calcium concentration in the medium. It is concluded that this large increase in passive "solubility" is caused by the diphosphonate's ability to stabilize the small amount of regulator phase (brushite or brushite-apatite mixture) present in bone. PMID- 6282410 TI - Bone response to phosphate and vitamin D metabolites in the hypophosphatemic male mouse. AB - The hypophosphatemic male mouse (Hyp/y), the proposed model for human vitamin D resistant rickets (VDRR), is characterized by chronic hypophosphatemia, dwarfism, and rachitic and osteomalacic bone lesions. We have reported that treatment of Hyp/y mice with phosphate salts (Pi) heals rickets but does not correct the defective endosteal bone mineralization. In an attempt to cure osteomalacia, mutant male animals were treated with Pi combined with 25-hydroxyvitamin D3 (25OHD3, 1 microgram/kg/day), 24,25-dihydroxyvitamin D3 [24,25(OH)2D3, 0.5 microgram/kg/day], or 1,25-dihydroxyvitamin D3 [1,25(OH)2D3, 0.05--0.25 microgram/kg/day] infused constantly for 3 weeks. The biochemical and skeletal effects of treatment were assessed by analytical methods and bone histomorphometry. The results show that only 1,25(OH)2D3 produced a dose dependent elevation of serum calcium and phosphorus, and greatly improved bone mineralization at doses high enough to increase serum calcium and phosphorus concentrations within or above the normal range. Better improvement of bone mineralization was obtained when Pi was combined to 1,25(OH)2D3. In conjunction with the correction of hypocalcemia, Pi + 1,25(OH)2D3 suppressed the stimulation of bone turnover induced by Pi supplementation. The results show that, as in VDRR children, 1,25(OH)2D3 produces beneficial effects on bone lesions in Hyp/y mice, mainly through enhancement of mineral availability. However, the persistence of osteomalacia despite correction of serum mineral concentrations suggests that there is a specific bone cell resistance to mineral and/or hormonal influences in Hyp/y mice. PMID- 6282412 TI - ESR and IR studies of carbonate-containing hydroxyapatites. AB - Synthetic carbonate-containing and carbonate-free hydroxyapatites(CO3-HA and CO3 free HA) have been investigated using ESR and IR absorption, together with X-ray diffraction and chemical analyses. Comparison of the IR spectra between 13C enriched 13CO3-HA and 12CO3-HA prepared at 100 degrees C and at high pH suggested that the carbonate substitutes for PO4 but not for HPO4. The substitution of CO3 for PO4 in this case resulted in elimination of hydroxyl ions in a hexad axis. The elimination of hydroxyl ions was found to be related to the decrease in crystallinity for the c-axis direction, although so far there has been no direct evidence for the existence of hydroxyl deficiencies. In the case of CO3-free HAs prepared at 80 degrees, 60 degrees, 40 degrees, and 20 degrees C, the decrease in crystallinity accompanied by the decrease in the temperature for preparation was associated with the decrease in crystal size, and no significant elimination of hydroxyl ions would be expected in the absence of the carbonate. PMID- 6282411 TI - Structural factors influencing the ability of compounds to inhibit hydroxyapatite formation. AB - For a compound to inhibit potently the transformation of amorphous calcium phosphate into hydroxyapatite, it is suggested that the minimum structural requirement is a phosphate group and, at some other position, either another phosphate group preferably or a carboxylic moiety. Primary amino groups abolish inhibitor potential. Inhibitor potency is modified by various secondary factors, including the number and proximity of active groups, their stereochemistry, steric factors, the lability of the molecule, and in special instances its lipophilicity. Parameters used to monitor the transformation indicate that inhibitors can be grouped into two classes, and it is suggested that this is because one class acts as a hydroxyapatite crystal growth inhibitor. The close proximity of two phosphate groups or of a phosphate and multiple carboxylic groups is proposed to determine in part whether or not a compound acts as a crystal growth inhibitor. Further, bulky side groups render a molecule inactive as a crystal growth poison, although it will still inhibit by other mechanisms. PMID- 6282413 TI - Calcitonin effects on isolated bone cells. AB - Calcitonin decreased calcium uptake in specific rat bone cell populations obtained by sequential collagenase digestions of calvaria. The calcitonin effect on calcium uptake was observed in the same populations that manifested calcitonin induced increases in cyclic AMP as well as high levels of acid phosphatase and the ability to release 45Ca from prelabeled devitalized bone. No effect of calcitonin was observed in cell populations that had high levels of alkaline phosphatase and lacked the potential to resorb devitalized bone. These results suggest that changes in cell calcium as well as cyclic AMP may be involved in the mode of action of calcitonin on osteoclast-like cells. PMID- 6282415 TI - Planar faults in dental hydroxy-apatite. PMID- 6282414 TI - Parathyroid hormone and calcitonin interactions in bone: irradiation-induced inhibition of escape in vitro. PMID- 6282416 TI - A Col E1 hybrid plasmid containing Escherichia coli genes complementing d-xylose negative mutants of Escherichia coli and Salmonella typhimurium. AB - The Clarke and Carbon bank of Col El - Escherichia coli DNa hybrid plasmids was screened for complementation of d-xylose negative mutants of E. coli. Of several obtained, the smallest, pRM10, was chosen for detailed study. Its size was 16 kilobases (kb) and that of the insert was 9.7 kg. By transformation or F' mediated conjugation this plasmid complemented mutants of E. coli defective in either D-xylose isomerase or D-xylulose kinase activity, or both. The activity of D-xylulose kinase in E. coli transformants which bear an intact chromosomal gene for this enzyme was greater than that for the host, due to a gene dosage effect. The plasmid also complemented D-xylose negative mutants of Salmonella typhimurium by F'-mediated conjugation between E. coli and S. typhimurium. Salmonella typhimurium mutants complemented were those for D-xylose isomerase and for D xylulose kinase in addition to pleiotropic D-xylose mutants which were defective in a regulatory gene of the D-xylose operon. In addition, the plasmid complemented the glyS mutation in E. coli and S. typhimurium. The glyS mutant of E. coli was temperature sensitive, indicating that the plasmid carried the structural gene for glycine synthetase. The glyS mutation in E. coli maps at 79 min, as do the xyl genes. The behaviour of the plasmid is consistent with the existence of a d-xylose operon in E. coli. The data also suggest that the plasmid carries three of the genes of this operon, specifically those for D-xylose isomerase, D-xylulose kinase, and a regulatory gene. PMID- 6282417 TI - The anaerobic sn-glycerol-3-phosphate dehydrogenase: cloning and expression of the glpA gene of Escherichia coli and identification of the glpA products. AB - The expression of recombinant plasmids carrying the glpA gene (anaerobic glycerol 3-phosphate dehydrogenase) and the closely linked glpT gene (glycerol 3-phosphate transport) were studied under aerobic and anaerobic conditions. When the pattern of expression of enzymatic activity in different strains was compared with sodium dodecyl sulphate - polyacrylamide gel electrophoresis (SDS-PAGE) analysis from the same cells the glpA products were identified. Two polypeptides of 62 000 and 43 000 relative mass correlated with enzymatic activity. Five recombinant plasmids that contained one or both of the glpT of glpA genes were isolated and subjected to restriction endonuclease cleavage analysis. The regions of overlap from the inserts in these plasmids allowed definition of the regions of DNA containing the glpT and glpA genes. SDS-PAGE analysis revealed a partial product of the glpA locus from one plasmid, pLC42-17, which allowed more precise definition of the glpA locus on the physical DNA map and prediction of the direction of transcription. PMID- 6282418 TI - Organization and expression of leghaemoglobin genes. AB - Leghaemoglobin genes in soybean (Glycine max) are present as a moderately reiterated family of sequences. Since there are identical restriction site patterns of these sequences in DNA isolated from leaf, root, or nodule tissue, the data suggest that no major changes in the organization or methylation of leghaemoglobin genes occur during their induction. Cloned soybean leghaemoglobin cDNA cross hybridized with RNA from root nodules of kidney bean (Phaseolus vulgaris), and to a lesser extent, of pea (Pisum sativum) indicating sequence homology in the leghaemoglobin genes of these species. Hybridization to the genomic DNA restriction fragments of two other species, Glycine soja and Vicia faba, also indicated interspecies sequence homologies. Several restriction fragments appear to be common to all species examined. The induction of these genes occurs following infection of the plant by Rhizobium and is independent of the appearance of nitrogenase activity in the nodules. The level of expression is, however, influenced by various mutations in Rhizobium that result in the development of ineffective (nonnitrogen fixing) nodules. PMID- 6282420 TI - On the contribution of the mitochondrial genome to the growth of Chinese hamster embryo cells in culture. AB - The present results demonstrate that Chinese hamster embryo cell populations in culture can be adapted to grow in the presence of chloramphenicol. It is shown that tryptose phosphate broth and uridine, one of its components, prevent the growth inhibitory effect of the drug. Study of some respiratory parameters (cytochrome c oxidase, cytochrome spectra, oxygen consumption) indicated that neither the broth nor uridine prevented the inhibitory effect of chloramphenicol on mitoribosomal protein synthesis. The cells grew with mitochondria devoid of a functional respiratory chain. Auxotrophy for pyrimidines appeared to result from the absence of dihydroorotate dehydrogenase, a respiratory chain-linked enzyme that catalyzes the fourth step of de novo pyrimidine biosynthesis. These and other results suggest that the synthesis of orotic acid may be considered as one of the main contributions of mitochondria to the growth of animal cells in culture. PMID- 6282419 TI - Potential of intense gamma-irradiation of host cells for analysis of virus specified transcription and replication. AB - Intense gamma-irradiation from a cobalt source differentially affects macromolecular synthesis of cultured mammalian cells. Exposure of monkey BSC-1 or murine fibroblastic L2 cells to 40 or 70 krad (1 rad = 1 x 10(-2) J/kg) abolishes DNA and RNA synthesis almost entirely but reduces the formation of protein much less. A dose-response analysis of irradiation shows that synthesis of total RNA and the messenger component thereof, measured as the poly(A)-containing fraction, are equally diminished. Host cells in which formation of DNA and RNA are minimal can support normal or nearly normal replication and transcription rates of vesicular stomatitis and vaccinia viruses. Therefore, use of pretreatment with gamma-irradiation, as employed here, should prove to be generally useful in examining virus-related transcription under circumstances in which application of drugs affecting gene expression, such as actinomycin D, is deemed undesirable. PMID- 6282421 TI - Chromosomal protein poly(ADP-ribosyl)ation in pancreatic nucleosomes. AB - When pancreatic chromatin fragments were prepared and resolved in the presence of 80 mM NaCl, endogenous poly(ADP-ribose) polymerase activity was found to be maximal in nucleosome periodicities of four to five units and did not respond to any further increases in nucleosomal architecture. Furthermore, in nucleosome complexities spanning 1 through 14 and over unit lengths, polyacrylamide gel electrophoresis on acid-urea and acid-urea-Triton gels has shown pancreatic histone H1 to be the only actively ADP-ribosylated histone species. The extent of ADP-ribosylation of histone H1 was also demonstrated to retard the protein's mobility in acid-urea, acid-urea-Triton, and lithium dodecyl sulfate polyacrylamide gels and to consist of at least 12 distinct ADP-ribosylated species extractable in all nucleosome complexities studied. Finally, extraction and subsequent electrophoresis of total chromosomal proteins in the presence of lithium dodecyl sulfate also evidenced heavy ADP-ribosylation at the level of nonhistone chromosomal proteins of the high mobility group comigrating in the core histone region, as well as in the topmost region of the gels where poly(ADP ribose) polymerase was found to form a poly(ADP-ribosyl)ated aggregate. PMID- 6282422 TI - Identification of gene products from cloned fragments of the left arm of lambda dapB2. AB - A set of recombinant plasmids encompassing the bacterial substitution in lambda dapB2 (Mackie, G. A. (1980) J. Biol. Chem. 255, 8928--8935) has been characterized genetically by complementation and biochemically by an analysis of the proteins encoded by individual plasmids in vitro. In addition to a cluster of four genes which includes the dapB locus, a total of seven gene products has been identified. One of these is ribosomal protein S20, whose gene (rpsT) has been localized to a 0.56-kilobase segment of DNA bounded by HindIII and HindII sites. Positive identification of this gene on plasmids pGM9 and pGP2 has been achieved by analysis of the products encoded by these plasmids in vitro and by the ability of pGM9 to complement a strain lacking S20 among its 30S subunit proteins. A second gene is that for isoleucyl tRNA synthetase (ileS). Its presence on pGM21 has been ascertained by the latter's ability to direct the synthesis of a protein in vitro with the size anticipated for this gene product. Extracts of cells harbouring this plasmid also exhibit greater isoleucyl tRNA synthetase activity than parental extracts. A third gene is at least 1.3 kilobases distant from rpsT and encodes a protein of 24 000 molecular weight, of unknown function. This locus is in turn about 6.5 kilobases from lambda gene E. The latter gene, and several others, all of which likely derive from lambda DNA are carried on a 5.2-kilobase fragment inserted in the plasmid pGM11 which includes the junction between phage and bacterial sequences on the left side of lambda dapB2. This fragment of DNA is inserted into the vector in a manner which permits the coupled transcription and translation of lambda genes D and E in vitro, despite the absence of their natural promoter. The most striking feature of the organization of the bacterial genes on lambda dapB2 is the clustering of genes for polypeptides in the right half of the bacterial substitution in contrast to the apparent minimal use of the coding potential in the left half of the substitution. PMID- 6282423 TI - DNA organization in nucleosomes. AB - A new technique known as electron spectroscopic imaging has allowed the direct visualization of DNA within the nucleosomes of chromatin. The results presented here confirm the model which suggests that approximately two supercoil turns of DNA are wound about the nucleosome core. The structure of nucleosomes from putative transcriptionally active genes, fractionated by preferential sensitivity to DNAase II and solubility in 2 mM MgCl2, has been examined using both dark field electron microscopy and electron spectroscopic imaging. Oligomeric strands of nucleosomes in this fraction have a less distinct beaded appearance than those of bulk chromatin. The phosphorus distribution in this chromatin suggests that the DNA has a less recognizable organization, lacking a two-turn supercoil per subunit. The unique appearance of this fraction in 30 mM NaCl is reversibly changed to the classical beaded appearance when dialyzed into 0.4 M NaCl. PMID- 6282425 TI - Factors affecting the survival of patients with lacrimal gland tumours. AB - This paper presents the clinical and pathological findings in 54 consecutive patients with primary tumours of the lacrimal gland. In the 30 cases of benign mixed-cell tumour the results of total surgical removal of the affected gland were good: no recurrences were noted during a follow-up period of up to 12 years. For patients with carcinoma of the lacrimal gland the outlook is generally poor, but 7 of the 24 patients in this series, all 7 of whom were treated surgically, have survived without recurrence for 5 years or more. The methods of treating these patients are described and the criteria used for choosing therapy discussed. PMID- 6282424 TI - DNA binding proteins from Tetrahymena thermophila. AB - We have used DNA-cellulose chromatography to isolate single-strand binding proteins from Tetrahymena thermophila. Three major proteins which bind to denatured DNA-cellulose were obtained. The predominant protein has a molecular weight of 20 000 in sodium dodecyl sulfate - polyacrylamide gel electrophoresis and possesses many of the properties of the helix destabilizing proteins isolated from prokaryotic and eukaryotic sources. The protein facilitates denaturation of the synthetic copolymer poly[d(A-T).d(A-T)], depressing the melting temperature by nearly 40 degrees C. It also permits the renaturation of poly[d(A-T)].d(A-T)] in high salt concentration. Two other binding proteins have molecular weight of 25 000 and 23 000 in sodium dodecyl sulfate - polyacrylamide gel electrophoresis. The protein with a molecular weight of 25 000 is probably the "M protein" previously isolated from Tetrahymena thermophila which has been shown to stimulate Tetrahymena DNA polymerase. These two proteins failed to show helix destabilizing, DNA dependent ATPase, or deoxyribonuclease activities. These three proteins are abundant in the cell with approximately 1.0 x 10(6) to 10.0 x 10(6) molecules of each protein monomer per cell. One molecule of each protein monomer binds to 7 to 10 nucleotides as detected by a nitrocellulose filter binding assay. Peptide mapping of the three proteins suggests that they are all distinct. We have also found that the binding proteins can interact with Tetrahymena DNA polymerase and some other proteins to form an enzyme complex, a putative replication complex. PMID- 6282426 TI - Blood pressure and cardiac tissue responses to prostacyclin (PGI2) in various species. AB - The effect of prostacyclin (PGI2) on blood pressure and heart rate (in vivo) and on isolated heart tissue has been investigated in different species. Isolated cardiac tissue had limited responses to PGI2 tested at 10(-13) to 10(-5) M. Cultured neonatal rat heart cells did not respond to PGI2, neither did intact rat hearts or rabbit cardiac tissue. Guinea pig and rat atria showed limited dose dependent responses to PGI2 at concentrations greater than 10(7) M. In rat atria, 10(-5) M PGI2 produced a limited elevation of tissue cAMP content. When given by intravenous injection or infusion, PGI2 produced hypotension in anaesthetized primates (three species), rat, rabbit, pig, and dog. As a vasodepressor in all species, PGI2 (on a weight basis) was more active than prostaglandins of the B or E type and, in most species tested, it was approximately five times more active than PGE2. Heart responses in intact animals were often paradoxical in that decreases in heart rate often accompanied blood pressure falls. PMID- 6282428 TI - Catabolite repression in Streptomyces venezuelae. Induction of beta galactosidase, chloramphenicol production, and intracellular cyclic adenosine 3',5'-monophosphate concentrations. AB - Chloramphenicol production was studied in cultures of Streptomyces venezuelae growing in a simple buffered medium with ammonia as the nitrogen source and glucose, lactose, or a glucose-lactose mixture as the sole source of carbon. With each carbon source the antibiotic was formed during growth. In the glucose lactose medium, the production pattern was biphasic; a marked decrease in the rate of synthesis was associated with depletion of glucose from the medium and a corresponding diauxie pause in growth. Cells of S. venezuelae contained an inducible beta-galactosidase. Induction by lactose was suppressed by glucose. Measurement of the concentration of intracellular adenosine 3',5'-cyclic monophosphate during growth of cultures with glucose or a glucose-lactose mixture as the source of carbon showed no appreciable changes coinciding with depletion of glucose or the onset of chloramphenicol biosynthesis. It is concluded that the cyclic nucleotide does not mediate selective nutrient utilization or control antibiotic biosynthesis in this organism. PMID- 6282427 TI - Baclofen-induced decrease of excitability of primary afferents and depression of monosynaptic transmission in cat spinal cord. AB - The relationship of the depressant effect of baclofen on spinal monosynaptic transmission and its effect on the excitability of primary afferents was examined in spinal unanesthetized cats. Baclofen (1.0 mg/kg, i.v.) produced a deep and long-lasting depression of spinal reflex responses with a concomitant decrease of terminal excitability. Primary afferent depolarization, as indicated by an increase of terminal excitability, evoked by conditioning of an antagonistic muscle nerve, was greatly reduced by this drug. Depression of monosynaptic transmission induced by baclofen was temporarily reversed by posttetanic potentiation. However, the same high frequency orthodromic stimulation further reduced excitability of terminals. It is therefore unlikely that block of terminal invasion is responsible for baclofen-induced depression of spinal monosynaptic transmission. These results are compatible with the suggestion that baclofen causes a reduction of transmitter release. In the spinal cord, this action is probably limited to the excitatory transmitter of primary afferents. PMID- 6282429 TI - Oncocytoma of the lung. AB - This report describes the clinical, anatomical, histologic and ultrastructural features of a bronchial oncocytoma. Histologically the oncocytes have granular eosinophilic cytoplasm. Ultrastructurally this corresponds to mitochondrial hyperplasia. Because cytoplasmic eosinophilia may be due to an abundance of cytoplasmic organelles other than mitochondria, it is emphasized that electron microscopic study is an essential component in the diagnosis of oncocytomas. PMID- 6282430 TI - Detection of adenoviruses in stools of children with nonbacterial gastroenteritis. PMID- 6282431 TI - Vesicular diseases: recent advances and concepts of control. AB - Most of this review is devoted to foot-and-mouth disease, considering only briefly vesicular stomatitis and vesicular exanthema of swine. PMID- 6282432 TI - A schirrhous anaplastic carcinoma in two dogs resembling mesothelioma. AB - The clinical, laboratory, radiographic and pathological findings of two dogs with similar neoplasms are described. Both dogs were mature males of a large breed and had pleural and abdominal effusions containing neoplastic cells. Diffuse scirrhous thickening of the pleura and serosal surfaces was a prominent feature. A fibrous layer containing nests of neoplastic, epithelial-like cells was present on serosal surfaces. The morphological appearance of the cells was suggestive of either an atypical sclerosing mesothelioma or a scirrhous carcinoma. On the basis of ultrastructural and differential staining characteristics, a diagnosis of a scirrhous carcinoma of undetermined primary origin was made in one case and of prostatic urethral origin in the other case. PMID- 6282433 TI - Epidemiology of breast carcinoma III: relationship of family history to tumor type. AB - Previous breast carcinoma in at least one female relative was reported by 31% of 1024 women treated consecutively for breast carcinoma at Memorial Hospital. Eighty (7.9%) of their mothers had had breast cancer. Maternal breast cancer was significantly (P less than 0.006) more frequent among women with medullary carcinoma than those with other tumor types and among those who were pre- or perimenopausal at diagnosis (P less than 0.001). Among the 727 patients who had one or more sisters, 12% had a sister who had been treated for breast cancer. The highest frequency of carcinoma in at least one sister occurred in patients with lobular carcinoma while the medullary carcinoma group had the least number of patients with an affected sister (P less than 0.03). Occurrence of breast cancer in a sister was almost twice as common in patients who were postmenopausal at diagnosis (P less than 0.005) than in premenopausal patients. When stratified by histologic type, the mean age at diagnosis of the patients did not differ appreciably from the age at diagnosis of their sisters. Detailed analyses of histologic type and other more distant familial relationships were also obtained but were considered to be less reliable because of problems in ascertainment and there were fewer affected relatives. No single histologic type of carcinoma was consistently linked to a disproportionately high or low frequency of carcinoma in all classes of relatives. It is possible that studies of family history limited to information available when the patient is first treated present an incomplete picture of familial aggregation. Further follow-up after diagnosis is needed to obtain a more reliable measure of the extent to which relatives are affected by breast cancer and patterns of family distribution associated with specific tumor types. PMID- 6282434 TI - Combination chemotherapy-radiotherapy with and without the methanol-extraction residue of Bacillus Calmette-Guerin (MER) in small cell carcinoma of the lung: a prospective randomized trial of the Piedmont Oncology Association. AB - The effect of addition of the nonspecific immunostimulant, MER, to combined treatment with chemotherapy and radiotherapy in small cell carcinoma of the lung was evaluated in a prospective randomized trial involving 102 evaluable patients. Chemotherapy consisted of cyclophosphamide, Adriamycin, vincristine, methotrexate, and CCNU; and radiotherapy was administered to the primary lesion, mediastinum, supraclavicular areas, and whole brain. Of 47 patients administered MER 400 mcg intradermally every six weeks, 12 (26%) attained complete remission with a median survival of 22.9 months. Complete remission was observed in 17 (31%) of 55 patients who received no MER with a median survival of 20.0 months (P greater than 0.05). Survival greater than or equal to 2 years has been observed in five patients who received MER and two patients who did not receive MER. The response rate and duration, survival, and toxicity of the two treatment arms were similar with the exception of cutaneous and occasional systemic reaction to MER. MER as used in this study has not influenced the overall results of a combined modality treatment program for patients with small cell carcinoma of the lung. PMID- 6282435 TI - Evaluation of 5'-nucleotide phosphodiesterase isozyme-V as a predictor for liver metastasis in breast cancer patients. AB - 5'-nucleotide phosphodiesterase isozyme-V (5'-NPD-V) was evaluated in 85 biopsy proven breast cancer patients as a potential marker for early liver metastasis. It correctly predicts liver metastasis in 6/7 (85.7%) patients with abnormal radiologic liver scan and 2/2 other patients with palpable liver. Serum glutamic oxaloacetic transaminase (SGOT), lactic dehydrogenase (LDH), alkaline phosphatase (AP) and total bilirubin (B) were also determined in 79 of these patients as routine liver function tests (LFT). Forty-one out of 79 from this group had all four markers all within normal limits. Yet of the 41 patients, 12 patients were found positive for 5'-NPD-V. Of these 12, one was found to have liver metastasis at surgery and one had abnormal liver scan. Five other patients had liver dysfunction and one had been diagnosed as an alcoholic. Four others had no evidence of either liver problems or liver metastasis, but follow-up data were lacking. This retrospective study, therefore suggests that there is a definite advantage to include the 5'-NPD-V in the liver profile studies for breast cancer patients, although a positive 5'-NPD-V may only indicate liver repair or liver regeneration. Long-term prospective studies of these tests with breast cancer patients should be worthwhile. No relation was found between 5'-NPV-V and axillary lymph node involvement or the estrogen receptor status of the excised tumor. Thus there is no evidence currently that the appearance of the 5'-NPD-V in serum is related to lymph node metastases or hormonal control. PMID- 6282436 TI - Computed tomography of malignant tumors of the nasal cavity and paranasal sinuses. AB - Staging of malignant tumors of the nasal cavity and paranasal sinuses by computed tomography (CT) was studied in a total of 49 patients, 33 with squamous cell carcinoma and 16 with tumors of other histologic types. Involved sites by the tumor were studied, and clinical staging was made using CT findings alone according to AJC classification for maxillary sinus tumors. Surgical findings for comparison were available for most cases. Of 33 squamous cell carcinomas and of 16 tumors with other histologic types, the maxillary sinus was the site of origin in 29 and eight, respectively. Of these 37 maxillary sinus tumors, 11 were staged T3, 26, T4, and none was staged T1 or T2. None of these tumors were down staged, and one T3 was upstaged after surgical procedures, although all sinuses were not explored in some cases. Sinusitis due to obstruction was indistinguishable from the tumor without bone destruction. And the determination of the site of origin was difficult in some cases. Despite these, CT should be used for pretreatment evaluation of the tumors of these sites. PMID- 6282437 TI - The immunohistochemical reactivity of a human monoclonal antibody with tissue sections of human mammary tumors. AB - Human monoclonal antibodies have been generated following the fusion of human lymphocytes (obtained from axillary lymph nodes of mastectomy patients) with a murine nonimmunoglobulin secreting myeloma cell line. We report a detailed analysis of the reactivity of a human IgM monoclonal antibody secreted by one of these double cloned hybridoma cell lines. Tissue sections of both malignant and benign human mammary tumors, as well as apparently normal tissues, were tested using the immunoperoxidase technique and the human monoclonal antibody. Eighty one per cent (54/67) of primary malignant mammary tumors, 100% (20/20) of metastatic breast lesions, and 14% (3/22) of benign breast lesions reacted positively with a moderate for strong intensity. The per cent of mammary carcinoma cells that stained and the pattern of staining varied among different tumor samples. While reactivity was observed with selected carcinomas of nonbreast origin, little or no reactivity was observed with apparently normal human tissues including normal mammary epithelium. The antibody reactivity observed was shown to be clearly distinct from those of both anti-T and anticarcinoembryonic antigen sera. PMID- 6282438 TI - Orofacial synovial sarcoma: a clinicopathologic study of 11 new cases and review of the literature. AB - Synovial sarcoma arises most commonly in the lower extremity, particularly in the region of the lower thigh and knee. Yet the occurrence of thus mesenchymal neoplasm in the head and neck area has been convincingly documented, albeit confined almost exclusively to cervical and parapharyngeal sites. Therefore, in view of its rarity in the head and neck, we analyzed a group of 11 synovial sarcomas arising in the orofacial region. The series comprised nine men (82%) and two women (18%). In common with synovial sarcoma at more conventional sites, this is a disease of young adults: ages ranged from 16-49 years (median, 34 years). Topographically, two subsets were delineated, a more common facial group with eight cases (four cheek, two parotid region, one infraorbital, one submental), all arising as gradually enlarging, usually nontender, solitary tumors; and three intraoral ones (two tonsillar, one lingual), two of which were polypoid and one was an exophytic tonsillar mass which presented with hemoptysis and stridor. Follow-up data, obtained for nine patients (range, 1.3-15.0 years), disclosed three (33%) tumor-related deaths, all belonging to the facial group. Treatment, difficult to significantly correlate with survival in this small series, varied from surgical excision alone to a multimodality approach including both irradiation and chemotherapy. Histologically, all the neoplasms revealed characteristic biphasic features, predominantly fibrosarcomatous in one, but, more typically, showing epithelial clefts and/or pseudoacinar spaces in the others. Differential diagnosis, depending on the proportion of the biphasic components, ranged from spindle cell mesenchymal neoplasms to various adenocarcinomas, including those arising in mixed tumors of salivary gland. PMID- 6282439 TI - Cerebellar glioblastoma. PMID- 6282440 TI - Spontaneous regression of hepatocellular carcinoma: a case study. AB - A Chinese patient with documented hepatocellular carcinoma (HCC) satisfied the criteria of Everson and Cole for spontaneous regression of malignant tumors. Subsequently he survived a tumor-free period of at least 13 years. During the period of regression, shrinkage of liver coincided with a rise of SGOT to a level comparable to that reported for patients with liver cancer during hepatic arterial ligation and cytotoxic therapy. Postregression liver biopsy from the site of the previous tumor revealed relatively uninflamed HBsAg-positive tissue without dysplasia. The case provided the positive end of the survival spectrum in HCC, evidence that regression of HCC might occur by involution rather than maturation, and histologic data suggesting that regressed HCC might be replaced by surrounding tissue instead of leaving behind dysplasia. PMID- 6282441 TI - Calcifying synovial sarcoma. AB - Analysis of the clinicopathological features and the available follow-up data in 32 patients with extensively calcified synovial sarcoma revealed an age incidence and anatomic distribution similar to that of ordinary synovial sarcomas without or with minimal calcification, but indicated a more favorable survival. The median age of 32 patient in this series was 26 years; the predominant location of the tumor was the soft tissues of the lower extremities; its average size was 4.6 cm (range, 1.5-20 cm). All of the tumors showed a focal biphasic cellular pattern and exhibited roentgenographic or microscopic evidence of extensive calcification and/or osseous metaplasia. Of the 26 patients with follow-up information (average 8.9 years) 17 were alive and well and six had died of their diseases, two after 7 and 19 years. Three additional patients were living with no signs of disease 1 1/2, 2 1/2, and three years after diagnosis, respectively. The five-year survival rate of 82.6 percent of this series is considerably better than the reported five year survival rates of synovial sarcoma which range from 25-51%. PMID- 6282442 TI - Genital warts and cervical cancer. I. Evidence of an association between subclinical papillomavirus infection and cervical malignancy. AB - A blind comparative survey was undertaken to study the prevalence of subclinical papillomavirus infection (SPI) in a representative sample of women treated surgically for invasive or preinvasive cervical neoplasia. According to a semiobjective rating system, 73 of 80 women (91%) with cervical neoplasia and ten of 80 matched controls (12.5%) showed histologic evidence of human papillomavirus (HPV) infection. Sixty of the controls (75%), but none of the study group, had normal cervicovaginal epithelium. A highly significant statistical relationship exists between subclinical papillomavirus infection of the lower genital tract and the occurrence of cervical neoplasia (F = 378; P less than 0.001; X2 = 109, P less than 0.001). The prevalence of SPI was seven times greater in the study group than in comparable controls of equivalent disease status. Because both are covariables of promiscuity, statistical association exist between cervical neoplasia and all sexually transmitted diseases. However, the strength, specificity and consistency of this relationship suggest that SPI may be a precursor or cervical malignancy. This contention is given biologic plausibility by a broad fabric of supporting epidemiology, virologic and clinicopathologic evidence. PMID- 6282443 TI - Growth of murine sarcoma and murine xenotropic leukemia viruses in Japanese quail: induction of tumors and development of continuous tumor cell lines. AB - The BALB/c murine endogenous xenotropic leukemia virus pseudotype of m1 Moloney sarcoma virus [m1MSV(B-MuX)] was used to productively transform diploid Japanese quail embryo cells. The majority of newly hatched quail inoculated with m1MSV(B MuX)-transformed quail embryo fibroblast cells rapidly developed tumors which were predominantly locally invasive fibrosarcomas, but metastases were observed in two birds. In two tumor-bearing quail, lymphosarcomas were observed in conjunction with fibrosarcomas. Quail inoculated with m1MSV(B-MuX) virus or quail embryo fibroblast cells infected with helper leukemia virus did not develop tumors. A cell culture derived from one quail tumor was shown to be oncogenic in newly hatched quail. Viruses produced by m1MSV(B-MuX)-infected quail cells or quail tumors had envelope properties of BALB-derived murine xenotropic leukemia virus as measured by host range, interference, and neutralization. Virus structural antigens, proteins with molecular weights of 30,000 and 70,000, were detected in tumors and tumor-derived cell lines by immunofluorescence and gel diffusion. Sera from tumored quail had high titers of type-specific BALB-derived murine xenotropic leukemia virus antibodies as determined by neutralization and immunoprecipitation. Antibodies to the putative m1 Moloney sarcoma virus mos gene product were not detected in sera from tumored or regressor quail. PMID- 6282445 TI - Similarities in the formation and removal of covalent DNA adducts in benzo(a)pyrene-treated BALB/3T3 variant cells with different induced transformation frequencies. AB - Classes of cell variants isolated from BALB/3T3-A31 clone 1 showing high, intermediate, and low susceptibility to ultraviolet light-induced transformation also demonstrated a differential response to benzo(a)pyrene (BP)-induced transformation. On the other hand, these variants showed an identical susceptibility to the killing effect of these carcinogens. The extent of BP binding to DNA was found to be very similar in all three classes of variants under transforming conditions. BP:DNA adducts from the variant cells were analyzed by high-pressure liquid chromatography. The same adducts (N2-(10S [7R,8S,9R-trihydroxy-7,8,9,10-tetrahydrobenzo(a)pyrene]yl)deoxyguanosine and a minute amount of the 7S enantiomer) were detected in the highly susceptible, the intermediately susceptible, and the resistant variant clones. Removal of the BP:DNA adduct occurred at an equally slow rate in all three classes of variant cells. These results indicate that the formation of stable covalent DNA adducts is not a sufficient condition for induction of transformation. Processes other than formation and removal of the stably bound BP:DNA covalent adducts are thus major factors controlling the susceptibility of BALB/3T3-A31 clone 1 variant cells to BP-induced transformation. PMID- 6282446 TI - Metabolic activation of benzo(a)pyrene by transformable and nontransformable C3H mouse fibroblasts in culture. AB - Incubation of 2.5 microM benzo(a)pyrene (BaP) with C3H/10T 1/2 or CVP3SC6 (CVP) mouse fibroblasts for 48 hr resulted in the metabolism of 36 to 42% of the BaP to organic soluble derivatives, which cochromatographed with 7,8-trans-dihydroxy-7,8 dihydrobenzo(a)pyrene, 9,10-trans-dihydroxy-9,10-dihydrobenzo(a)pyrene, 3 hydroxybenzo(a)pyrene, and 9-hydroxybenzo(a)pyrene, or to water-soluble derivatives. The formation of both organic and water-soluble metabolites during the 48-hr period increased proportionally with time, except in the case of BaP phenols, which increased initially but then remained the same or decreased. The distribution of organic soluble metabolites in the extracellular culture medium consisted primarily of BaP diols and was significantly different from that found inside the cells. The intracellular profile of organic soluble metabolites produced by both cell lines consisted predominantly of BaP phenolic derivatives and was qualitatively similar to the spectrum of metabolites produced by the incubation of BaP with C3H/10T 1/2 or CVP cell microsomes. The nature of the BaP water-soluble derivatives produced by the C3H/10T 1/2 and CVP cell lines was investigated by hydrolysis of culture medium with beta-glucuronidase and arylsulfatase. Although sulfation was not a major conjugation pathway for BaP in these cells, glucuronidation of BaP phenols was found to account for 30% of the total water-soluble derivatives. The similarity in the kinetics and qualitative nature of the metabolism of BaP by C3H/10T 1/2 and CVP cells indicates that both cell lines are equally capable of biosynthesizing the proximal carcinogen, 7,8 trans-dihydroxy-7,8-dihydrobenzo(a)pyrene. Analysis of the water-soluble metabolites produced by these cells suggests further that the nonresponsiveness of the CVP cells to BaP-induced transformation cannot be accounted for on the basis of an increased detoxication of 7,8-trans-dihydroxy-7,8 dihydrobenzo(a)pyrene. PMID- 6282444 TI - Benzo(a)pyrene metabolism in primary cultures of mouse epidermal cells and untransformed and transformed epidermal cell lines. AB - The metabolism of [3H]benzo(a)pyrene [B(a)P] by cultures of primary mouse epidermal cells and untransformed and transformed epidermal cell lines was investigated. All three cell types effectively metabolized [3H]B(a)P. The major organic solvent-extractable metabolites found intracellularly in primary cultures were trans-7,8-dihydro-7,8-dihydroxybenzo(a)pyrene and 3-hydroxybenzo(a)pyrene, although quantities of 9-hydroxybenzo(a)pyrene, trans-9,10-dihydro-9,10 dihydroxybenzo(a)pyrene, and quinones also were present. The major organic solvent-soluble metabolites found in the extracellular medium were trans-9,10 dihydro-9,10-dihydroxybenzo(a)pyrene and trans-7,8-dihydro-7,8 dihydroxybenzo(a)pyrene, with smaller quantities of unconjugated phenols and quinones. The major water-soluble metabolites found in the extracellular medium were conjugated with glucuronic acid [primarily 3-hydroxybenzo(a)pyrene and several quinones]. No sulfate conjugates of [3H]B(a)P metabolites were detected. [3H]B(a)P metabolism was similar in cultures of untransformed and transformed epidermal cell lines but differed from the primary cultures. The major intracellular and extracellular organic solvent-soluble metabolites were diols. Little or no unconjugated phenols were detected. Both the untransformed and transformed epidermal cell lines converted [3H]B(a)P to water-soluble metabolites, primarily glucuronide conjugates. In contrast to the primary cells, a major pathway of trans-7,8-dihydro-7,8-dihydroxybenzo(a)pyrene metabolism in the untransformed and transformed cell lines was a glucuronide conjugate. Primary mouse epidermal cells provide an important model system for studying factors affecting the activation and detoxification of hydrocarbon carcinogens. PMID- 6282447 TI - Electron spin resonance studies on intact cells and isolated lipid droplets from fatty acid-modified L1210 murine leukemia. AB - It has been suggested that the formation of cytoplasmic lipid droplets may produce an artifact and be responsible for the differences in membrane physical properties detected in lipid-modified cells using fluorescence polarization or spin label probes. To investigate this, the electron spin resonance spectra of lipid droplets isolated from the cytoplasm of L1210 leukemia cells were compared with spectra obtained from the intact cell. Mice bearing the L1210 leukemia were fed diets containing either 16% sunflower oil or 16% coconut oil in order to modify the fatty acid composition of the tumor. A microsome-rich fraction prepared from L1210 cells grown in animals fed the sunflower oil-rich diet contained more polyenoic fatty acids (52 versus 29%), while microsomes from L1210 cells grown in animals fed the coconut oil-rich diets contained more monoenoic fatty acids (37 versus 12%). The order parameter calculated for lipid droplets labeled with the 5-nitroxystearic acid spin probe was only about one-half that of intact cells, whereas it was similar to that obtained for pure triolein droplets suspended in buffer. Order parameters of the inner hyperfine splittings calculated from the spectra of cells grown in the sunflower oil-fed animals [0.543 +/- 0.001 (S.E.)] were lower than those from the cells grown in animals fed the coconut oil diets (0.555 +/- 0.002) (p less than 0.005). In contrast, the order parameters of the lipid droplets isolated from the cells grown in animals fed sunflower oil (0.303 +/- 0.029) or coconut oil (0.295 +/- 0.021) were not significantly different, indicating that motion of a spin label probe in the highly fluid cytoplasmic lipid droplets is not affected by these types of modifications in cellular fatty acid composition. Therefore, the electron spin resonance changes that are observed in the intact cells cannot be due to localization of the probe in cytoplasmic lipid droplets. These results support the conclusion that the electron spin resonance changes observed with the 5 nitroxystearic acid spin probe are due to changes in membrane fluidity produced by the modification in cellular lipid composition. PMID- 6282448 TI - Morphological, immunological, and biochemical analyses of chicken spleen cells transformed in vitro by reticuloendotheliosis virus strain T. AB - Reticuloendotheliosis virus strain T (REV-T) is a highly oncogenic avian retrovirus which causes a rapid neoplastic disease of the lymphoreticular system. We derived six cell lines (1-3, 1-5, 2-10, 2-14, 2-16, and 2-20) from chicken spleen cells infected with REV-T. These cells can produce both the REV-T and its associated reticuloendotheliosis helper virus, REV-A. Histochemical analyses of these cells indicate that, while they are not stained by benzidine, peroxidase, beta-glucuronidase or acid alpha-naphthyl acetate esterase, they contain a high proportion (95%) of cells positive for acid phosphatase. Light and electron microscopic studies of these cells also revealed morphologies of lymphoblasts or activated lymphocytes with irregular nuclei and dispersed chromatin. Immunochemical analyses indicate that essentially all (90 to 100%) of the cells contain the surface marker Ia, but no cytoplasmic immunoglobulin M and immunoglobulin G could be detected by immunofluorescence staining. Results also show that some of these cell lines contain a low level of terminal transferase (0.02 to 0.17 unit/10(9) cells), and a proportion (3 to 35%) of these cells can be stained by an antiserum directed against chicken bursa cells. These results are consistent with the conclusion that the cells transformed by the highly oncogenic REV-T are lymphoid in nature. In addition, at least some of these cell clones may contain features characteristic of activated B-lymphocytes. Analysis of these cell clones indicates that some cell lines contain an adherent and nonadherent population with some differences in morphologies. In addition, electron microscopic examination revealed that, while the non-adherent cells are actively producing type C viruses, type C viruses are either absent or very rare in the adherent cell populations. These results support the conclusion that some of these cell lines are heterogeneous and contain subpopulations of cells with differences in their ability to produce viruses. PMID- 6282450 TI - Interaction of benzo(a)pyrene and its dihydrodiol-epoxide derivative with nuclear and mitochondrial DNA in C3H10T 1/2 cell cultures. AB - To analyze the distribution of radioactive carcinogens and [3H]thymidine between nuclear DNA (nDNA) and mitochondrial DNA (mtDNA), we have developed a simple and rapid method for the separation of nDNA and mtDNA using gel electrophoresis of cell lysates. Using this method, we found that, when C3H10T 1/2 cells are exposed to either 0.5 microM [3H]-(+/-)-7 beta,8 alpha-dihydroxy-9 alpha,10 alpha-epoxy 7,8,9,10-tetrahydrobenzo(a)pyrene ([3H]BPDE) or 1 microM [3H]benzo(a)pyrene, the mtDNA contains a major fraction of the total adducts formed with cellular DNAs. Deoxynucleoside adducts formed between benzo(a)pyrene and mtDNA in intact C3H10T 1/2 cells or between BPDE and isolated rat liver mtDNA were analyzed by high performance liquid chromatography, and were found to be much more heterogeneous than those present in nDNA of C3H10T 1/2 cells. The extensive modification of mtDNA in BPDE in C3H10T 1/2 cells is associated with preferential inhibition of the incorporation of [3H]thymidine into mtDNA, when compared to incorporation of [3H]thymidine into nDNA. To analyze the factors responsible for the extensive modification of mtDNA by BPDE, we investigated the role of a lipid phase utilizing liposome:DNA complexes as a model system. We found that the liposomes protect BPDE from spontaneous hydrolysis and enhance the extent of DNA modification at low DNA concentrations. These findings extent previous evidence suggesting that the mitochondria may be important cellular targets in the process of chemical carcinogenesis. PMID- 6282449 TI - Cyclic adenosine 3':5'-monophosphate-dependent protein phosphorylation and the control of leukemia L1210 cell growth. AB - Synergistic increases in the survival of mice bearing an L1210 leukemia tumor have been demonstrated previously after treatment with 1,3-bis(2-chloroethyl)-1 nitrosourea together with theophylline over those treated with either agent alone. These results imply that manipulation of cyclic adenosine 3':5' monophosphate (cyclic AMP) levels in L1210 cells may result in alteration of sensitivity to chemotherapy and alterations in tumor growth. In the present study, we have shown that in vivo treatment of L1210 cells with theophylline results in changes in intracellular cyclic AMP-dependent protein kinase activity levels as well as in an apparent redistribution of both the nuclear and cytoplasmic isozymes. Biochemical events in the tumor cells immediately after administration of theophylline in vivo or a cyclic AMP analog (8 parachlorophenylthio cyclic adenosine 3':5'-monophosphate in vitro were independent of the presence of 1,3-bis(2-chloroethyl)-1-nitrosourea. The changes apparently involve signal transduction via the adenylate cyclase system and manifest as: (a) increased sensitivity of cyclic AMP-dependent protein kinase to activation by cyclic AMP after treatment of L1210 cells with theophylline; (b) decrease in endogenous nuclear protein phosphorylation sites; and (c) protein kinase isozyme redistribution between nuclear and extranuclear compartments, i.e., a relative increase of the type I isozyme activity in the nuclear and of the type II isozyme activity in the 900 x g supernatant fractions after treatment of the mice with theophylline. The relative activity increases are accompanied by a relative decrease of type II activity from the nucleus and type I isozyme activity from the 900 x g extranuclear supernatant fraction. These events appear temporally related to changes in nuclear RNA metabolism as evidenced by altered kinetics of RNA precursor uptake and incorporation into tumor cell RNA after treatment. These results imply that the cyclic AMP-dependent phosphorylative modification of intracellular proteins may play a regulatory role in tumor cell growth and in theophylline-mediated tumor regression. PMID- 6282452 TI - Urinary cyclic nucleotides and the cytopathology of human uterine cervical dysplasias. AB - Blinded urinary assays for cyclic guanosine 3':5'-monophosphate (cGMP) and cyclic adenosine 3':5'-monophosphate (cAMP) were performed on 49 subjects with documented abnormal cervical cytology and 21 control subjects with normal cytology. A significant difference in the mean cGMP:cAMP ratios between the case and control groups was found. A significantly greater proportion of women with cytological abnormalities had a cGMP:cAMP ratio above the 0.2 level (p less than 0.001). Cases treated surgically for severe dysplasia or carcinoma in situ of the cervix revealed a significant postsurgical fall in the cGMP:cAMP ratios (p less than 0.025). The possibility of utilizing urinary ratios of cyclic nucleotides as an objective index in the detection, monitoring of progression, and therapy of preneoplastic cervical lesions is discussed. PMID- 6282451 TI - Glycosaminoglycans in 3'-methyl-4-dimethylaminoazobenzene-induced rat hepatic cancer. AB - The changes in glycosaminoglycans in livers of rats with 3'-methyl-4 dimethylaminoazobenzene-induced hepatic cancer were examined and compared with those in fetal liver. The incorporation of 35S into sulfated glycosaminoglycans in hepatic cancer tissue was also studied after i.p. injection of Na(2)35SO4. The major component of glycosaminoglycans in healthy adult rat liver was heparan sulfate (61.7%), with hyaluronic acid (21.1%), dermatan sulfate (13.1%), and chondroitin sulfate (4.0%) as minor components. The quantities of all of the examined glycosaminoglycans were higher in tumors than in normal liver, but chondroitin sulfate and hyaluronic acid were more prevalent in the tumors (about 51 and 7 times higher, respectively). As a result, the share of heparan sulfate was decreased (37.1%) in the tumors. The high content of chondroitin sulfate (about 30 times) and the decreased share of heparan sulfate (29.0%) were also observed in fetal liver. The incorporation of 35S into individual glycosaminoglycans varied markedly. Approximately 90% of the label in glycosaminoglycans of healthy liver was found in a heparan sulfate fraction 4 hr after injection. In hepatic cancer tissue, however, 35S incorporation into both chondroitin sulfate and dermatan sulfate fractions was increased about 6.5- and 5.6-fold, respectively, of those in healthy liver. PMID- 6282453 TI - Lack of efficacy of high-dose methotrexate by 30-hour infusion in patients with progressive small cell carcinoma of the lung. AB - Seventeen patients with small cell carcinoma of the lung (SCCL) refractory to at least one combination chemotherapy regimen were treated with a high-dose methotrexate (HDMTX) program consisting of 1500 mg/m2 of MTX by iv infusion at a constant rate over 30 hrs following a priming dose of 50 mg/m2 of MTX iv. Administration of calcium leucovorin was begun at the end of the infusion. Treatment was repeated every 2 weeks. No complete or partial responses were observed, and the median time to tumor progression was 4 weeks. Hematologic toxicity was much more severe than anticipated, with five patients having thrombocytopenic bleeding requiring platelet transfusions. This HDMTX regimen was both ineffective and unexpectedly toxic in our heavily pretreated patients with SCCL. Literature review discloses little evidence that HDMTX programs have superior activity to standard doses of MTX in this disease. PMID- 6282454 TI - Hexamethylmelamine and cisplatin in advanced ovarian cancer after failure of alkylating-agent therapy. AB - Thirty-eight women with advanced ovarian cancer resistant to alkylating-agent chemotherapy were treated with a combination of hexamethylmelamine and cisplatin. Hematologic toxicity was severe or life-threatening in seven of 13 patients treated with cisplatin at a dose of 100 mg/m2 but was acceptable in 25 patients treated at 75 mg/m2. Thirty-four percent of the combined group developed peripheral neuropathy and only 8% of those treated at 75 mg/m2 developed transient azotemia. Nine complete responses and 12 partial remissions were noted among 38 patients treated, with no obvious advantage for the higher dose of cisplatin. The median duration of remission was 8 months and the overall median survival was 9 months. The response rate of 55% is approximately twice that reported with either agent given alone in similar patient populations and is equal to or better than that reported for more complicated regimens containing these drugs together with other, less active drugs. PMID- 6282456 TI - Prevention and reversal by a retinoid of 3,4-benzpyrene- and cigarette smoke condensate-induced hyperplasia and metaplasia of rodent respiratory epithelia in organ culture. AB - The influence of an aromatic analog of vitamin A, etretinate, on the effects of 3,4-benzpyrene and cigarette smoke condensate has been investigated in fetal mouse lung and neonatal rat tracheas grown in organ culture. In both tissues, 3,4 benzpyrene as well as cigarette smoke condensate induces a striking increase of epithelial mitosis within 12--14 days of treatment. The increase is associated with a loss of secretory activity and of ciliary function. These changes persist in the absence of benzpyrene or smoke condensate in explants transferred to control medium. Treatment with etretinate alone does not affect the normal epithelial growth rate or normal differentiation. If combined with either benzpyrene or smoke condensate, the aromatic compound inhibits the increase in cell division and prevents the loss of secretory activity or ciliary function. In explants pretreated with 3,4-benzpyrene or cigarette smoke condensate, etretinate reduces the carcinogen- or smoke condensate-induced increase in mitotic activity to normal levels and restores secretory differentiation and ciliary function. The mechanism of action involved in the anticarcinogenic activity of the retinoid is discussed. PMID- 6282455 TI - Chemotherapy versus chemoimmunotherapy with levamisole or Corynebacterium parvum in advanced lung cancer. AB - A total of 109 patients with advanced lung cancer, all cell types, were randomized between MACC chemotherapy only, consisting of methotrexate, doxorubicin (Adriamycin), cyclophosphamide, and lomustine (CCNU); MACC plus levamisole (LMS) orally; and MACC plus Corynebacterium parvum (CP) sc. Of these patients, 101 were evaluable, with no differences among the three treatment groups for overall response rate and survival time. Objective response rates and median survival times were 41% and 230 days for patients given MACC only, 39% and 257 days for those given MACC plus LMS, and 44% and 223 days for those given MACC plus CP, respectively. There was a significant increase in survival for patients with large cell anaplastic carcinoma receiving CP or LMS, particularly in the good-performance-status category. Pretreatment delayed cutaneous hypersensitivity to recall antigens in 50 patients had prognostic significance, but repeat tests after 2 months of treatment in 30 patients did not show different patterns of conversion among the three groups. There was no difference in hematologic toxicity among the three groups. With the possible exception of large cell anaplastic carcinoma, immunotherapy with LMS or CP as given in this trial does not appear to be therapeutically advantageous in advanced lung cancer. PMID- 6282457 TI - Workshop on Kaposi's sarcoma: meeting report. PMID- 6282458 TI - The epidemic of acquired immune dysfunction in homosexual men and its sequelae- opportunistic infections, Kaposi's sarcoma, and other malignancies: an update and interpretation. PMID- 6282459 TI - Non-cross-resistant chemotherapy and consolidation radiotherapy for small cell carcinoma of the lung. AB - Fifty-six patients with small cell carcinoma of the lung were treated with a two cyclic induction course of hexamethylmelamine, vincristine, doxorubicin, and cyclophosphamide. Patients with limited disease (LD) who responded and patients with extensive disease (ED) who had a complete response received prophylactic whole-brain radiotherapy, as well as radiotherapy to thoracic and abdominal sites of disease. Concurrently with radiotherapy, consolidation chemotherapy was given with doxorubicin, cyclophosphamide, methotrexate, and etoposide. The complete response rate was 35% for ED patients and 68% for LD patients. The median survival time for complete responders was 54 weeks for ED patients and 65 weeks for LD patients. The toxicity of the program was moderate, and the effectiveness was comparable to that of other reported combined-modality treatment programs. PMID- 6282460 TI - Phase II study of cisplatin, maytansine, and chlorozotocin in small cell lung carcinoma (EST 2578). AB - Seventy-three patients with small cell lung carcinoma refractory to standard chemotherapy were entered in this phase II randomized study of cisplatin, maytansine, and chlorozotocin. Of the 58 evaluable patients, only one partial response was observed among 21 patients given cisplatin, and no responses were seen among 19 given maytansine or 18 given chlorozotocin. One patient treated with chlorozotocin and two treated with cisplatin experienced life-threatening thrombocytopenia. One third of the maytansine-treated patients experienced moderate or severe neurologic toxicity. The overall median survival was 9.7 weeks. Chlorozotocin treatment was associated with inferior survival (7.7 weeks). PMID- 6282461 TI - Localization of gelfoam emboli after partial splenic embolization utilizing 99mTc labeled emboli. AB - Partial splenic embolization using 99mTc-labeled Gelfoam was employed to manage of four renal transplant patients with thrombocytopenia and leukopenia. In each patient the individual Gelfoam particles could be seen on the persistence scope of a gamma camera as they arrived in the spleen. This allowed the precise location of each Gelfoam particle to be defined immediately. Gamma camera imaging of the 99mTc Gelfoam particles in the spleen proved as accurate as selective splenic arteriography in determining the amount of spleen embolized. No ectopic 99mTc Gelfoam particles were found as judged by total body gamma camera imaging obtained immediately after the partial splenic embolization procedure was completed. PMID- 6282463 TI - Induction and catabolite repression of alpha-galactosidase from saccharomyces carlsbergensis. PMID- 6282462 TI - Pseudo-obstruction of the interior vena cava in pediatric abdominal masses. AB - Extrinsic compression of the inferior vena cava by large abdominal masses may simulate occlusion on supine cavography. Seventeen children with large abdominal masses were evaluated by inferior venacavography. There was either occlusion or severe compression of the IVC on supine cavography in 13 patients; but the decubitus cavagram and subsequent surgery confirmed patency in 12 patients. In four patients only minor indentations or no abnormalities were present. Pseudo obstruction of the IVC can be accurately differentiated from occlusion due to invasion by a lateral cavagram in the decubitus position. PMID- 6282464 TI - Regulation of amino acid transport into Ehrlich cells. Effect of 3',5'-adenosine monophosphate on L-leucine transport. PMID- 6282465 TI - Cyclic AMP-dependent protein kinase and endogenous phosphorylation levels in young and adult rat hearts. PMID- 6282466 TI - Modification of protein kinase assay for rapid and economic measurements. PMID- 6282467 TI - Metabolic-morphologic characteristics of the integument of teleost fish with mature lymphocystis nodules. AB - Normal and virus-infected (lymphocystis disease) integument from five species of teleosts was examined by light and TEM autoradiography and SEM to establish metabolic-morphologic characteristics of integument with mature lymphocystis cells (LC's). LC's with numerous morphologic attributes of a late developmental stage showed highest incorporation of [3H]-thymidine in vivo (1-91 h) above the intracytoplasmic inclusion body (ci) with little radiolabel in nuclei, cytoplasmic icosahedral deoxyriboviruses (ICDV's) or capsule. Analysis by quantitative autoradiography revealed that the % total cell label in ci and cytoplasm did not vary appreciably from 1-91 h and was corroborative with morphologic criteria of maturity. A possibly phylogenetic difference was noted between teleosts, wherein normal integument showed uptake of [3H]-thymidine in vivo (1 h) by cells at all levels of the epidermis, and cyclostomes (Spitzer et al. 1979) wherein labeling was confined to the basal third of the epidermis. Among four infected teleost species, the mean diameters of the ICDV's measured under the same conditions, ranged from 259.5 nm to 290.0 nm with the mean for each species differing significantly (p less than 0.01) from each of the other means. Ruptured LC's were shown by TEM and SEM to have released ICDV's onto the lesions and integument. Various stages of LC degeneration, host response, and integumental repair processes were documented. An evaluation of labeling in vivo of the capsular matrix was compatible ([3H]-D-galactose greater than [3H]-L lysine much greater than [3H]-L-fucose) with a glycosaminoglycan-protein structure. PMID- 6282469 TI - Macrophages as effector cells of protective immunity in murine schistosomiasis. PMID- 6282468 TI - Assessment of left ventricular function by first-pass radionuclide angiography in patients with poor left ventricular function. PMID- 6282470 TI - [Norwalk-like viruses as probable etiological agents in the epidemic of non bacterial gastroenteritis (author's transl)]. PMID- 6282471 TI - Adenosine 3',5'-cyclic monophosphate dependent and independent protein kinase activities in 1,2-dimethylhydrazine induced rat colon cancer. AB - The adenosine 3',5'-cyclic monophosphate (cAMP)-dependent and cAMP-independent kinase activities were measured in the 1,2-dimethylhydrazine (DMH) induced rat colon cancer and in untreated colon. Previous studies had shown that intestinal tumors induced by chronic exposure to DMH contained 2-fold less intracellular cAMP. The present findings indicate that reduction in cAMP-dependent protein kinase activities also occur in colon cancer cells. Similar hydrogen ion dependence (pH 6-7) and approximate association constants (Ka approximately 0.1 microM) were observed for the enzymes existing in both normal and tumor tissues, while the cAMP-dependent tumor protein kinase was found to phosphorylate phosvitin and casein to a greater degree. These recent findings are consistent with the concept that the concentrations of cAMP and activities of its associated enzyme system are inversely related to the cell proliferation state. PMID- 6282472 TI - Uptake of hexavalent chromium by bovine erythrocytes and its interaction with cytoplasmic components; the role of glutathione. AB - Hexavalent chromium (Cr(VI)) anion gradually penetrated into bovine erythrocytes and bound with cytoplasmic components. Its penetration was strongly inhibited by the NH2-reactive agent, 4-acetamido-4'-isothiocyano-stilbene-2,2'-disulfonic acid (SITS) and the SH-reactive agent, N-ethylmaleimide (NEM). Gel filtration showed that the intracellular component that bound to chromium was hemoglobin. The binding affinity of Cr(VI) to hemoglobin in the absence of glutathione in vitro was found to be much less than in intact erythrocytes. However, in the presence of glutathione, the binding affinity of Cr(VI) to hemoglobin became much higher. This indicates that reduction of hemoglobin or Cr(VI) by glutathione is involved in the binding. Cr(VI) interacted only weakly with the membrane and did not cause hemolysis of bovine erythrocytes, unlike heavy metals such as Hg2+. PMID- 6282473 TI - Time course of metabolism of benzo[e]pyrene by hamster embryo cells and the effect of chemical modifiers. AB - In cultures of hamster embryo cells, benzo[a]pyrene (B[a]P) is metabolized primarily in the bay region. In contrast, little or no bay region metabolism of the noncarcinogenic isomer benzo[e]pyrene (B[e]P) could be detected during 12--96 h incubations of hamster embryo cells with 4 microM [3H]B[e]P. The upper limit to 9,10-dihydro-9,10-dihydroxy-B[e]P formation is about 0.2% of the ethyl acetate soluble metabolites (less than 0.1% of the total metabolites). The major identified metabolites of B[e]P were 4,5-dihydro-4,5-dihydroxy B[e]P and the glucuronide conjugates of 3-OH-B[e]P and 4,5-dihydro-4,5-dihydroxy B[e]P. Simultaneous treatment of cells with either B[a]P or 7,8-benzoflavone (BF) did not induce bay region metabolism of [3H]B[e]P. PMID- 6282474 TI - Rat liver microsomal metabolites of 7-methylbenz[c]acridine. AB - The metabolism of the weak carcinogen 7-methylbenz[c]acridine (7MBAC) was examined in rat liver microsomes from 3-methylcholanthrene(MC)-induced animals by the use of mixed 14C- and 2H-labelled substrate. The three metabolites identified by spectroscopic and chromatographic examination were 7-OHMBAC and two dihydrodiols. The dihydrodiols were assigned structures consisted with attack on the 8,9- and 5,6- or K-region of the aromatic system. PMID- 6282475 TI - Characteristics of microsomal reduction of benzo[a]pyrene 4,5-oxide. AB - NADPH-reduction of benzo[a]pyrene 4,5-oxide (BP-4,5-oxide) to BP required four components from rat liver: cytochrome P-450, NADPH cytochrome P-450 reductase, phosphatidylcholine and a soluble, heat-sensitive factor which was present in 105 000 X g supernatant and was also released from microsomes by sonication. The requirement for this factor contrasts with recently reported results from Sugiura et al. (Cancer Res., 40 (1980) 2910). Oxide-reduction was 40 times faster under anaerobic conditions, but oxygen did not affect the stimulation factor. This stimulation was highest (X15) at low concentrations of microsomal protein (less than 0.1 mg/ml) and was almost absent at high concentrations of microsomal protein (greater than 1 mg/ml). Oxide-reduction activity was proportional to microsomal protein concentration in the presence of added 105 000 X g supernatant, but for microsomes alone (greater than 0.1 mg/ml) exhibited a parallel plot with an intercept at 0.08 mg/ml microsomal protein. Stimulation was highest at high concentrations of BP-4,5-oxide and a linear plot of V-1 vs. [BP 4,5-oxide]-1 was only obtained in the presence of 105 000 X g supernatant (Km = 3 microM, Vmax = 3.3 nmol/mg/min). Microsomal hydration of BP-4,5-oxide (inhibited in reductase assays) was unaffected by 105 000 X g supernatant, suggesting that stimulation of oxide-reduction did not derive from solubilization of BP-4,5 oxide. Stimulation was observed in the initial rate of reaction and was independent of incubation time. Inhibition of lipid peroxidation, removal of peroxides and deoxygenation were all excluded as explanations of the stimulatory effect. PMID- 6282476 TI - [A case of glomus tumor of the prepatellar region]. PMID- 6282477 TI - [World epidemiological situation: viral disease incidence (author's transl)]. AB - The importance of viral disease, although estimated with difficulty, seems very large in human pathology. Viruses appear to be involved in most of respiratory disease. Moreover, in tropical countries, measles and viral diarrhoeas (mainly rotaviruses) are fatal for many children. But smallpox eradication must be considered as the most important fact which occurred in the field during the last five years, and the consequences of this new situation are discussed. Finally, beyond the recent question of african viral haemorrhagic fevers, one must take in account the increasing prevalence of several arboviruses, specially dengue in southeast Asia, which is one of the first causes of paediatric mortality in urban environment. PMID- 6282479 TI - Viral agents of acute respiratory infections in young children in Kuala Lumpur. AB - The results of this study indicate that the important viral agents associated with lower respiratory tract infections in young children are respiratory syncytial virus, rhinovirus, and parainfluenza virus, particularly in those under 2 years of age. This is in close agreement with studies done in temperate climates. Influenza A virus is seasonal and plays an important role in upper respiratory tract infections in older children. PMID- 6282478 TI - Rotavirus and acute diarrhoeal disease in children in a southern Indian coastal town. AB - Rotavirus was found by electron microscopy in the stools of 70.7% of a representative sample (368) of the 3355 children with acute diarrhoea admitted to hospital over a period of 16 months in Calicut on the west coast of India. The prevalence of the virus was high (nearly 100% of cases examined) in the period from November to January and lowest in May just before the onset of the monsoon. Prevalence was high (75.1%) in infants aged from 6 to 23 months, but was considerably lower in those under 6 months of age (34.8%). The management of cases and the planning of control measures for this disease are discussed in the light of knowledge of the high prevalence of rotavirus. PMID- 6282480 TI - Epipodophyllotoxin (VP16-213) in small cell carcinoma of the Bronchus resistant to initial combination chemotherapy. AB - Thirty-eight patients with small cell carcinoma of the bronchus resistant to initial chemotherapy with cyclophosphamide methotrexate and CCNU, were treated with VP16-213 alone in a dose of 120 mg/m2 i.v. on days 1, 3, and 5 every 3 weeks. Twelve patients died before three courses of treatment. In 26 patients who received three or more courses only one, transient partial response occurred. One or more components of the initial chemotherapy seems to confer resistance to the action of VP16-213 in this disease. PMID- 6282481 TI - Treatment of small cell lung cancer with a combination of VP16-213 and cyclophosphamide with cisplatin or radiotherapy. AB - Fifty five patients with small cell lung cancer with treated with VP16-213 combination chemotherapy regimen in two consecutive series. The first series included 24 patients; 10 with limited and 14 with extensive disease were treated with VP16-213, 120 mg/m2 p.o. daily for 5 consecutive days, Cyclophosphamide 600 mg/m2 i.v. and Cisplatin 80 mg/m2 i.v. with hydratation and manitol induced diuresis. The cycle was repeated every 3 weeks. The second series included 31 similar patients, 16 limited, and 15 extensive disease, treated with VP16-213 at the same dose and Cyclophosphamide at 1,200 mg/m2 i.v. also repeated every 3 weeks; after three cycles the patients were treated with radiotherapy to the primary tumor and regional lymph nodes with 4,000 rads in a split course of three weeks interval, followed by the same combination chemotherapy. Response rate was 75% for the first series with 6 of 24 (25%) of complete responses in four limited and two extensive disease and a median survival time of 24 weeks. In the second series of patients there were 26 of 31 (83.8%) responses with 10 of 31 (32%) complete responses in nine limited and one extensive disease and a median survival time of 33 weeks for responders. Duration of response for complete responders was 36.8 weeks for the first series and 51 weeks for the second. Toxicity was mild and includes nausea and vomiting, myelosupression, alopecia in both series, with one toxic death in the second series. Both regimens are active with a low complete response rate, which was increased in the second series by the addition of radiotherapy, which did not increase overall survival. PMID- 6282483 TI - Vincristine, Adriamycin, cyclophosphamide, and etoposide (VP16-213) in small-cell anaplastic carcinoma of the lung. AB - Twelve patients with small-cell anaplastic carcinoma of the lung were treated with vincristine 1 mg/m2 i.v. day 1, adriamycin 50 mg/M2 i.v. day 1, cyclophosphamide 1,000 mg/m2 i.v. day 1, and etoposide 80 mg/m2 i.v. day 2, 4, 6 given on an outpatient basis and repeated at 3-week intervals. As consolidation therapy seven patients received two courses of BCNU median 29 mg/m2 (range 24-73 mg/m2) short term intraarterial infusion in the bronchial artery with 2 to 3 weeks intervals. One patient with limited disease had no evidence of disease for 13+ months and one patient complete remission for 3+ months. Four of ten patients with extensive disease had complete remission for median 5 months (range 2+ to 5+ months) and four patients had partial remission for median 5 months (range 4 to 5+ months). Despite side effects the chemotherapy was well tolerated by the patients. The results correspond to those obtained with other effective regimens in small-cell anaplastic carcinoma of the lung. PMID- 6282482 TI - Doxorubicin, Cyclophosphamide and VP16-213 (ACE) in the treatment of small cell lung cancer. AB - Small cell lung cancer requires aggressive combination chemotherapy. The three active agents, doxorubicin (A) 45 mg/m2 i.v. day 1, cyclophosphamide (C) 1.0 mg/m2 i.v. day 1 and VP16-213 (E) 50 mg/m2/day i.v. days 1-5 were given together. The combination (ACE) was given every 21 days without chest irradiation. One hundred and seventy-four patients have been stratified for extent of disease and randomized on three sequential studies testing ACE vs ACE + MER immunotherapy (38 patients), or ACE vs ACE alternating with CCNU, methotrexate, vincristine and procarbazine (109 patients), or ACE vs ACE II (ACE with continuous VP16-213 - 100 mg/m2/day X 5 days - 27 patients - ongoing). The immunotherapy and the alternating non-cross resistant combination have not proven beneficial with respect to response or survival. The ACE combination, regardless of additional treatments, has produced greater than 90% response overall. In limited disease the complete response (CR) frequency is 65%. The median survival for limited disease overall is 14 months and 18 months for patients achieving CR. In extensive disease the CR frequency is 40% with a median survival of 9 months overall and 13 months for patients achieving CR. Response frequency and survival are identical in the first two studies and 20-30% of patients with limited disease are long-term survivors with one late relapse (greater than 3 years). Patients who achieved CR had a significantly longer survival regardless of other factors such as performance status or extent of disease. Prophylactic cranial irradiation was demonstrated to be useful in prevention or delaying CNS metastases in patients who achieved CR. The third generation study of high-dose VP16-213 infusion seeks to increase the CR frequency. ACE chemotherapy without chest irradiation is a highly effective treatment for all patients with small cell lung cancer and compares favorably with all other studies with or without adjuvant radiotherapy. PMID- 6282484 TI - Chemotherapy combination with cyclophosphamide (CTX) adriamycin (ADM), vincristine (VCR), and VP16-213 in small cell carcinoma of the lung (SCCL). AB - Twenty-four evaluable patients with small cell carcinoma of the lung were treated with an escalating chemotherapy regimen including Cyclophosphamide, Adriamycin, Vincristine and VP16-213. The initial doses were CTX 800 mg/m2 i.v. day 1; ADR 50 mg/m2 i.v. day 1; VCR 1.4 mg/m2 day 1 weekly; and VP16-213 100 mg/m1 i.v. days 14 18 every 4 weeks, CTX and ADR were escalated by 100 and 10 mg/m2 respectively in each subsequent cycle according to blood count. Hematologic toxicity was minimal and the treatment was well tolerated. Partial responses and complete responses were 9 of 19 and 5 of 19 respectively for patients with limited disease, and 4 of 5 respectively for patients with extensive disease. The overall response rate for the whole group was 79% These results must be considered preliminary. PMID- 6282485 TI - VP16-213 combined with cis-platinum (CDDP) in the treatment of small cell carcinoma of the lung (SCLC). PMID- 6282486 TI - VP16-213, cisplatinum, and adriamycin salvage therapy of refractory and/or recurrent nonseminomatous germ cell neoplasms. AB - Eleven patients with recurrent and/or resistant nonseminomatous germinal cell neoplasms refractory to conventional chemotherapy were treated with the combination. VP16-213, cis-diamminedichloroplatinum, and adriamycin. One complete response, four partial responses which at surgery were benign teratomas, and six partial responses have been observed. Four patients are prolonged survivors (greater than 18 months). The possibility that this regimen may offer true salvage for refractory patients exists. Incorporation of VP16-213 into initial treatment regimens for germinal cell neoplasms is warranted. PMID- 6282487 TI - The formation of benzo [a] pyrene - deoxyribonucleoside adducts in vivo and in vitro. AB - The formation of DNA-bound products from benzo [a]-pyrene was studied using metabolic activation systems of widely differing cellular and structural integrity. The nature of the benzo [a] pyrene-metabolite-deoxyribonucleoside adducts formed was markedly dependent on the nature of the activation system using. systems where cellular integrity is preserved, namely mouse skin in vivo and rat liver hepatocytes and lung explants in vitro, when treated with benzo [a] pyrene, formed deoxyribonucleoside adducts of which the major one co chromatographed with a bay-region diol epoxide-deoxyguanosine adduct, 10 beta (deoxyguanosin-N2-yl)-7 beta, 8 alpha, 9 alpha-trihydroxy-7,8,9,10 tetrahydrobenzo-[a] pyrene. In striking contrast, the use of activating systems with disrupted cellular integrity such as rat liver, rat lung and mouse skin microsomal fractions, in the presence of benzo [a] pyrene and exogenous DNA, resulted in the formation of deoxyribonucleoside adducts derived predominantly from either a further metabolite of 9-hydroxybenzo [a] pyrene or (+/-) benzo [a] pyrene-4,5-epoxide. The consequences of such results are important when considering the widespread use of subcellular fractions such as microsomes as metabolic activating systems for short-term mutagenicity and carcinogenicity tests. PMID- 6282488 TI - Carcinogenicity of 2-amino-4-(5-nitro-2-furyl) thiazole in rats by oral and subcutaneous administration. AB - Female Fischer rats were divided into two groups and fed either basal diet or basal diet containing 0.166% 2-amino-4-(5-nitro-2-furyl) thiazole for 52 weeks followed by basal diet for an additional 16 weeks. Our of the 24 rats fed 2-amino 4-(5-nitro-2-furyl) thiazole, 23 had forestomach tumors, four had fibroadenomas and two had adenocarcinomas in the mammary glands, two had renal pelvic carcinomas, five had papillomas and six had carcinomas in the bladder, and two had cutaneous tumors. None of the 10 rats fed basal diet developed tumors. In another study, two-week-old Fischer rats of both sexes were injected s.c. once a week for the first eight weeks with 2-amino-4-(5-nitro-2-furyl) thiazole at a dose of 10 mg/kg and then 1.4 mg/rat for an additional 46 weeks. The control rats were injected with solvent (dimethyl sulfoxide: water, 1:1) only. The experiment was terminated at the end of 58 weeks. Malignant fibrous histiocytomas were found in five out of eight male rats and four out of 11 female rats, and an angiosarcoma was found in one out of eight male rats injected with 2-amino-4-(5 nitro-2-furyl) thiazole. None of the control rats developed tumors. PMID- 6282489 TI - Mechanism for the loss of preferential benzo [a] pyrene binding to the linker DNA of chromatin. AB - We have examined the fate of the asymmetric chromosomal distribution of DNA adducts generated by the chemical carcinogen r-7,t-8-dihydroxy-t-9,10-oxy 7,8,9,10-tetrahydro-benzo[a]pyrene (BPDE). Treatment of mouse embryo cells with BPDE results in 3.5 times more binding to the linker DNA regions between nucleosome cores than to the nucleosome core DNA itself, but 24 h post-treatment incubation of these cells leads to a loss of this non-random binding. A similar result was obtained when post-treatment incubation was carried out in the presence of hydroxyurea indicating that factors other than DNA replication are responsible for this changes in adduct distribution. However in the case of excision repair deficient xeroderma pigmentosum (XP12/BE) cells the non-random adduct distribution was stable over a period of 48 h, whereas with excision repair proficient XP variant (XP4/BE cells, loss of preferential binding did occur. There results indicate that the loss of non-random nucleosomal DNA modification with time can be accounted for by the preferential removal of adducts from micrococcal nuclease sensitive linker DNA and further, demonstrates that in certain cells at least, the relative position of nucleosome core structures on DNA remains unchanged over a period of at least 48 h. PMID- 6282490 TI - Indirect immunofluorescent localization of benzo[a]pyrene adducted to nucleic acids in cultured mouse keratinocyte nuclei. AB - The localization of benzo[a]pyrene-deoxyguanosine adducts was studied by indirect immunofluorescence in cultured BALB/c epidermal cells exposed to (+/-) 7 alpha, 8 alpha-dihydroxy-9 alpha, 10 alpha-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene (the anti-isomer) utilizing an antiserum specific for the major benzo[a]pyrene deoxyguanosine adduct in DNA. This antiserum does not cross-react with benzo[a]pyrene or DNA alone. When cultured keratinocytes were incubated with the carcinogen for 1 h, the immunofluorescence was localized in the nucleus as intense spots on a background of diffuse fluorescence. Fluorescence was absent from cells not exposed to carcinogen and from carcinogen-exposed cells incubated with normal rabbit serum in place of the antiserum. Fluorescence was abolished when the specific antiserum was absorbed with the immunogen DNA prior to incubation with cells, and substantially diminished when exposed cells were preincubated with deoxyribonuclease before the application of the specific antiserum. Incubation of exposed cells with ribonuclease prior to incubation with the specific antiserum removed the bright fluorescent spots and resulted in fluorescent nuclei containing dark sports in similar frequency. Dose-response studies in which benzo[a]pyrene-deoxyguanosine adducts were quantified by enzyme linked immunosorbent assay and compared with intensity of immunofluorescence demonstrated that decreasing doses of the carcinogen resulted in fewer numbers of adducts as well as proportionally less fluorescence. When cells were exposed to non-toxic doses of the activated carcinogen for 1 h, nuclear fluorescence was detectable in immediately-fixed cells but faded to non-detectable levels when cells were washed and cultured for an additional 24-48 h before fixation. PMID- 6282491 TI - Contraction-excitation feedback in myocardium. Physiological basis and clinical relevance. PMID- 6282492 TI - High force development and crossbridge attachment in smooth muscle from swine carotid arteries. AB - In experiments designed to achieve maximal activation, the active force/cell cross-sectional area in tissues prepared from the swine carotid media was 6.7 +/- 0.3 (sd) X 10(5) N/m5. This value exceeds that reported for other vertebrate muscle cells and is striking because of the low smooth muscle myosin content. The hypothesis that high force generation may, in part, reflect an increase in the crossbridge duty cycle, i.e., the fraction of the cycle during which force is generated, was tested by determining the rate of force redevelopment after a step shortening and the ration of the load-bearing capacity of the contractile system to the developed stress during the course of isometric contractions. Maximal crossbridge cycling rates estimated by the rate of force redevelopment occurred 30 seconds after the onset of a high K+-induced contraction, and decreased thereafter, although the load-bearing capacity or maximum active stress was maintained. These results from isometric experiments support the hypothesis and provide further evidence that attached, non-cycling crossbridges contribute to force maintenance in tonically contracting arterial smooth muscle. PMID- 6282493 TI - Increased vascular catecholamine sensitivity and alpha-adrenergic receptor affinity in female and estrogen-treated male rats. PMID- 6282494 TI - Juxtaglomerular cells grown as monolayer cell culture contain renin, angiotensin I-converting enzyme, and angiotensin I and II/III. AB - A monolayer cell culture of juxtaglomerular cells (JGC) was derived from the renal cortex of neonatal rats. The JGC had the characteristics of those within the kidney, including peripheral dense bodies and myofibrils indicating a smooth muscle origin; rough ER containing fluffy material consistent with protein synthesis; a prominent Golgi apparatus for packaging granules, and granules having the characteristics of secretory granules and lysosomes. Transplants of the cultured cells into syngeneic recipients survived for 2 weeks or longer and retained the features of JGC. The JGC granules fluoresced when treated with a rabbit antibody against pure rat renin, followed by fluorescein isothyocyanate conjugated F(ab')2 fragment of goat antirabbit IgG (Fc fragment) heavy chain specific. The latter indicated the presence of renin. The JGC were lysed in the presence of DFP, captopril, leupeptin, and EDTA, and were extracted in the presence of pepstatin. The lysate contained renin activity that was inhibited by a specific renin antibody. Nonspecific proteases were excluded by the antibody and its pH optimum. Angiotensin I-converting enzyme was detected in the lysate prepared without the use of EDTA and captopril. Angiotensins I and II/III were derived from the extract by additional extractions, TLC, and RIA, using highly specific antibodies. The angiotensins were confirmed by chromatography monitored by authentic angiotensins. We concluded that the cultured JGC contained renin, angiotensin I-converting enzyme, and angiotensin I and II/III. PMID- 6282496 TI - Intercellular communication in cardiac muscle. PMID- 6282495 TI - Effect of histamine on the energy metabolism of K+-depolarized hog carotid artery. AB - In a previous study (Peterson and Gluck, 1982), activation of hog carotid artery with high-K+ medium caused no increase in aerobic glycolysis (lactic acid production). In contrast, histamine alone is found to stimulate aerobic glycolysis in a tension-dependent fashion (GLuck and Paul, 1977). In this study, the addition of histamine to the already K+-depolarized hog carotid artery induced additional contraction, elevated tissue cyclic adenosine 3',5' monophosphate [cAMP] levels, coupled the rate of aerobic glycolysis to external [Ca++] and isometric tension, and caused suprabasal energy utilization to exceed that required to support the contraction alone by approximately 25%. Increased cytoplasmic free-[Ca++] per se does not stimulate aerobic glycolysis. In the presence of histamine, however, aerobic glycolysis is Ca++-sensitive. Whereas a cycle of Ca++ depletion and restoration has only a small effect on basal metabolic rates in arteries activated with high-K+ alone, the same procedure with arteries stimulated by high-K+ with added histamine causes a dramatic shift in basal metabolic rates toward higher levels of glycolysis. PMID- 6282498 TI - Influence of heat stress on arterial baroreflex control of heart rate in the baboon. AB - The influence of environmental heat stress on the arterial baroreflex control of heart rate (HR) was studied in eight conscious, chronically instrumented baboons. Inflations of balloon occluders around the inferior vena cava (IVC) and thoracic descending aorta (DA) were used to produce acute, graded changes in mean arterial blood pressure (MABP) in 5 mm Hg intervals ranging from +/- 5 to +/- 25 mm Hg. After determination of the HR responses to changes in MABP in the normothermic baboon (blood temperature less than or equal to 37.6 degrees C), the animal was subjected to environmental heating to produce hyperthermia. When blood temperature reached approximately 39.5 degrees C, HR responses to graded DA and IVC occlusions were again determined. During hyperthermia, the HR sensitivity (delta HR/ delta MABP) to MABP changes was markedly diminished for reductions in MABP and significantly enhanced for increases in MABP. To determine whether these alterations in the HR response to changes in MABP were due to an alteration of the baroreflex control of HR, full, sigmoid-shaped HR-MABP curves for both the normothermic and hyperthermic states were constructed and characterized by total HR range, estimated slope of the steep portion of the curve, and MABP at the midpoint of the HR range (BP50). During hyperthermia (1) the whole HR-MABP curve shifted significantly upward by 35-40 beats/min, (2) total HR range, the estimated slope, and BP50 did not change, and (3) the control point (pre occlusion HR-MABP value) curves were also constructed during either beta adrenergic blockade or cholinergic (Ch)-receptor blockade in the normothermic and hyperthermic state. Similar to that seen for the unblocked heart, the whole HR MABP curves were also shifted upward during hyperthermia in this group of baboons with no alteration in the total HR range, the estimated slope, or BP50. The upward shift in the HR-MABP curve during Ch-receptor blockade, unlike during beta receptor blockade, was much greater than that which could be attributed only to the local effect of blood temperature. Although the control point was also shifted upward along the steep portion of the curve during beta- or Ch-receptor blockade, the upward shift observed during beta-adrenergic blockade was similar to that observed in the unblocked state. Thus, a heat stress-induced hyperthermia produces a rise in HR without significantly altering the characteristics of the reflex control of HR by arterial baroreceptors. To rely solely on changes in HR sensitivity may lead to erroneous conclusions as to the effect of a particular stress on the baroreceptor reflex control of HR. Further, these results indicate that: (1) the upward shift in the HR-MABP curve is mediated by both the local effect of blood temperature on HR and cardiac sympathetic efferent neurons which are independent of the baroreceptor reflex, and (2) the upward shift in the control point is mediated predominantly by vagal withdrawal, probably as part of the compensatory response to a heat-induced hypotension. PMID- 6282497 TI - Regulation of large coronary arteries by beta-adrenergic mechanisms in the conscious dog. AB - We examined, in conscious dogs, the effects of beta-adrenergic stimulation on measurements of left circumflex coronary arterial diameter and blood flow and on calculations of late diastolic coronary resistance (LDCR) and left circumflex coronary internal cross-sectional area (CSA). Isoproterenol (0.1 microgram/kg) initially decreased mean arterial pressure by 25 +/- 2% (mean +/- SEM), and LDCR by 62 +/- 4%, and increased heart rate by 82 +/- 10%, left ventricular (LV) dP/dt by 79 +/- 12%, and mean coronary blood flow by 85 +/- 5%, while CSA rose slightly. The peak effects on CSA (24 +/- 2%) occurred later, along with decreases in mean arterial pressure (7.4 +/- 1.0%) and LDCR (25 +/-5.3%) and increases in coronary blood flow (14 +/- 2%), LV dP/dt (12 +/- 3%), and heart rate (24 +/- 4%). Pirbuterol (1.0 microgram/kg) induced changes that were qualitatively similar to those induced by isoproterenol. Prenalterol (20 micrograms/kg), a cardioselective beta 1-adrenergic receptor agonist, did not affect mean arterial pressure, but increased heart rate by 40 +/- 5%, LV dP/dt by 72 +/- 10%, mean coronary blood flow by 34 +/- 11%, and CSA by 26 +/- 3%, and decreased LDCR by 29 +/- 5+. Isoproterenol and pirbuterol, but not prenalterol, increased coronary sinus O2 content and decreased A-VO2 difference. After beta 1 adrenergic receptor blockade with atenolol (1 mg/kg), prenalterol no longer induced significant effects, whereas isoproterenol and pirbuterol decreased mean arterial pressure similarly to what was observed prior to blockade, but did not increase LV dP/dt, and induced attenuated increases in mean coronary blood flow, CSA, and decreases in LDCR. Thus, in the intact, conscious animal, large coronary arteries are regulated by beta-adrenergic mechanisms. Surprisingly, a major fraction of large coronary arterial dilation appeared to be either directly or indirectly due to beta 1-adrenergic receptor mechanisms, although beta 2 adrenergic effects were also significant. PMID- 6282500 TI - An organic phosphorus assay which avoids the use of hazardous perchloric acid. PMID- 6282499 TI - Possible role for cytotoxic oxygen metabolites in the pathogenesis of cardiac ischemic injury. AB - Parabiotically perfused, isolated cat hearts were subjected to global ischemia for 2 hours at 27 degrees C and were then reperfused for 2 hours. One group of hearts was simply perfused with hypothermic blood 5 minutes before ischemia and again before normothermic reperfusion. A second group received a crystalloid cardioplegic solution. A third group received cardioplegic solution supplemented with superoxide dismutase and catalase; at the start of reperfusion the anesthetized cats supplying blood to perfused hearts in this group received an i.v. infusion of these enzymes. Based on comparisons of postreperfusion ventricular pressure development, maximal ventricular dP/dt, rate-pressure products and coronary blood flow, we concluded that cardioplegic solution and enzyme treatment were superior to enzyme-free cardioplegia solution under otherwise similar experimental conditions. The results suggest that a component of "ischemic" cardiac damage may involve cytotoxicity from oxygen metabolites such as superoxide anion, hydrogen peroxide, or both, and that this component of damage can be reduced by enzyme supplements such as superoxide dismutase or catalase. PMID- 6282501 TI - Assay, kinetics and properties of plasma adenosine diphosphatase. The relationship to acid and alkaline phosphatase and variations in disease. AB - A rapid radioassay was used to characterise the adenosine diphosphatase (ADPase) activities in human plasma. There was a major peak at pH 9.3, 80% of whose activity was attributable to non-specific alkaline phosphatase, with the remaining 20% probably due to a specific ADPase. There was also a small peak of ADPase activity at pH 4.0. Inhibitor and chromatographic studies showed that whilst much of this activity was attributable to non-specific acid phosphatase, there was a discrete acid ADPase. Assays of plasma ADPase activities in vascular disorders, including myocardial infarction, peripheral vascular disease and diabetes mellitus, reveal no alterations from control values. Activities of alkaline ADPase were elevated in both chronic and acute liver failure. Acid ADPase was also increased in chronic liver disease and it is suggested that alterations in ADPase activities in liver disorders may contribute to the haemostatic problems observed in these patients. PMID- 6282503 TI - [A case of insulinoma studied on secretion of gastric inhibitory polypeptide (author's transl)]. PMID- 6282502 TI - The decreased basal and stimulated prolactin levels in isolated gonadotrophin deficiency: a consequence of the low oestrogen state. PMID- 6282504 TI - [A study on pregnancy-specific beta 1-globulin assay (SP1-RIA) and its diagnostic application (author's transl)]. PMID- 6282505 TI - The treatment of patients with chronic mucocutaneous candidosis and candida onychomycosis with ketoconazole. PMID- 6282506 TI - Detection and characterization of circulating immune complexes during acute exacerbation of chronic viral hepatitis. AB - For detection and characterization of circulating immune complexes (CIC) in various liver diseases, a C1q binding test (C1q BT) was used. While the CIC level was almost normal in HB surface antigen (HBsAg) positive asymptomatic carriers, it was relatively high in patients with liver diseases. The study of the sedimentation rate of CIC in various liver diseases showed two kinds of CIC; one greater than the other. During acute exacerbation of chronic active liver diseases, the CIC level reached its peak 1-5 weeks before and after the peak of sGPT. In acid buffer, CIC in one patient with HBsAg positive severe chronic aggressive hepatitis was dissociated into 5-6 fractions by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The two of these fractions proved to be HBsAg and IgG and another three or four undetermined components. At the acute exacerbation period of this case, the fraction pattern of CIC by SDS-PAGE was similar to each other at any of the four stages and HBsAg always composed one of the antigens. PMID- 6282507 TI - Complement receptor enhancement and chemotaxis of human neutrophils and eosinophils by leukotrienes and other lipoxygenase products. AB - The lipoxygenase products of arachidonic acid, 5-HPETE, 5-HETE, LTB4, LTC4 and LTD4, were examined for their capacity to enhance the expression of complement (C3b) receptors and to evoke chemotaxis of human neutrophils and eosinophils. With the exception of LTD4 all gave enhancement of C3b receptors. LTB4 and LTC4 enhanced over the concentration range 10(-7) to 10(-11) moles/l, and 5-HETE and 5 HPETE from 5 X 10(-6) to 5 X 10(-10) moles/l. The rank order of activity, as assessed by the magnitude of enhancement, was LTB4 (concentration for maximal effect = 10(-7) moles/l) greater than 5-HETE (5 X 10(-7)) greater than 5-HPETE (5 X 10(-6)) greater than LTC4 (10(-9)). High dose inhibition was observed with LTC4 and 5-HETE. Chemotaxis experiments performed in parallel over the same concentration ranges indicated that neither neutrophils nor eosinophils migrated towards LTC4 or LTD4. However, LTB4 evoked chemotaxis with a linear dose response from 10(-9) to 10(-7) moles/l and 5-HPETE and 5-HETE from 5 X 10(-8) to 5 X 10( 6) moles/l. At 10(-7) moles/l LTB4 was approximately 6 and 8 X more potent in chemotaxis than 5-HPETE and 5-HETE respectively. In general, complement receptor enhancement and chemotaxis of eosinophils were similar to that observed with neutrophils and did not vary with the patient source. PMID- 6282510 TI - Mitral valve prolapse and partial syndactyly of the hands. AB - A patient is presented who has mitral valve prolapse and syndactyly of the hands. This combination of findings has not been previously reported, but is consistent with one theory of the development of mitral valve prolapse. PMID- 6282508 TI - Immunological profile of amyotrophic lateral sclerosis patients and their cell mediated immune responses to viral and CNS antigens. AB - The 'immunological profile' of amyotrophic lateral sclerosis (ALS) patients was established from standard tests for B- and T-cell function. This showed no significant difference from age and sex-matched other neurological (CNS) disease controls and normal subjects. Immune complex (IC) levels in ALS serum differed significantly from normal controls but not from CNS controls. There was no relation between the various indices of immune activity of IC levels and the clinical disability of the ALS patient or progression of the disease. Distribution of complement-fixing antibodies to poliovirus was similar to sera of ALS and control groups. The in vitro cell-mediated immune responses to poliovirus, however, were significantly greater in ALS patients than in CNS controls and were inversely related to the ALS disability score. Poliovirus has not been demonstrated in the CNS or extra-CNS tissues of ALS patients by conventional means but, if latent or defective poliovirus or related virus were present, this could account for sensitization and a possible autoimmune mechanism. ALS patients exhibited in vitro cellular immunity to ALS and normal CNS subfractions. These responses were not related to the ALS disability score or progression of the disease and probably represent epiphenomena. PMID- 6282511 TI - Bone cell membranes. AB - While many, if not most, of the details remain to be worked out, bone cell function is increasingly being explained in terms of cell membrane phenomena. Ultrastructurally, bone cell membranes do not show remarkable features beyond those associated with the ruffled border and clear zone of the osteoclast or specializations associated with the formation of matrix vesicles. Major physiologic properties of bone cell membranes are related to calcium transport, electrical properties and hormonal responses. Major advances in these areas are occurring with the development of separation techniques for the isolation of subpopulations of bone cells that respond primarily to parathyroid hormone or calcitonin. The combined actions of calcium and cyclic AMP in large part mediate the hormonal and possible electrical responses of bone cells. PMID- 6282509 TI - The effects of T-cell growth factor and virus purification on virus-mediated inhibition of lymphocyte mitogenesis. AB - Many different types of virus particles are able to non-specifically impede the ability of human peripheral blood lymphocytes to respond to mitogenic ro alloantigenic stimuli. This result is not obtained if ultra-purified virus is employed, although virus which has been banded only once through sucrose generally retains inhibitory potential. Ultra-pure virus is relatively unable to bind to cell surfaces, suggesting the importance of physical contact between viruses and cells in order for the observed inhibition to occur. Addition of exogenous T-cell growth factor (TCGF) to cultures containing virus, cells and stimulus causes a dose-dependent reversal of the usual inhibitory effect. PMID- 6282512 TI - The pattern of recovery after severe head injury. AB - The pattern of recovery after head injury was studied on the basis of clinical data on 200 patients. An unmistakable pattern was found in which it was possible to distinguish between symptoms of only short duration (due to the unfavourable prognosis, among other things), symptoms which showed a fairly quick recovery such as the E score and the oculovestibular response, and symptoms characterized by a longer interval between the accident and their recovery. A clear difference was found between the pattern recovery of E scores and oculovestibular response on the one hand, and recovery of M and V scores on the other. The implications of the pattern of recovery thus established, are discussed. PMID- 6282513 TI - Dystonia musculorum deformans with hyperuricaemia - study of a family. AB - This article presents a study of a family in which association of hyperuricaemia and more interestingly increased C.S.F. uric acid was noticed in two members who had classical dystonia musculorum deformans (DMD). Other members of the family had no clinical evidence of DMD but had increased serum and C.S.F. uric acid. Investigations did not reveal any of the common disorders of the purine metabolism in any of the family members. An attempt has been made to discuss the aetiopathological correlation. PMID- 6282514 TI - Success and failure of evoked potentials in detecting clinical and subclinical lesions in multiple sclerosis patients. AB - The value of visual, brain stem auditory and somatosensory potentials in detecting clinical and subclinical lesions as compared to the routine neurological, ophthalmological and vestibular examinations was investigated in 100 M.S. patients. It would appear that the VEP and SEP are far superior to the routine techniques in demonstrating lesions. On the other hand, the BAEP is inferior to the clinical and vestibular test as an indicator of brain stem lesions. All clinically manifest posterior column lesions are associated with abnormal SEP. However a substantial proportion of clinically evident lesions in the visual pathways or the midbrain and pons are not detectable by the VEP and BAEP. PMID- 6282515 TI - Transverse myelitis associated with mycoplasma pneumoniae infection. PMID- 6282516 TI - Cerebral metastases from pulmonary and mammary carcinomas. AB - The prognosis of patients subjected to operation for a solitary cerebral metastasis from a pulmonary carcinoma is unfavourable. The prognosis of a solitary cerebral metastasis from a mammary carcinoma that is removed appears to be better: a prolongation of life of acceptable duration and quality is frequently achieved. PMID- 6282517 TI - Migraine-like headache in intraventricular tumours. AB - In a series of 39 intraventricular tumours, 7 patients had paroxysmal headache as the most important early complaint. The striking similarities with migraine and the misleading role of antimigraine drug therapy are illustrated. Special emphasis is placed on the importance of additional investigations such as funduscopy, EEG, skin X-rays and Ct-scan. PMID- 6282518 TI - Phycomycosis of the C.N.S. first report of a Dutch case. PMID- 6282519 TI - Adverse side effects on cerebral functions after the use of metrizamide for cervical myelography. PMID- 6282520 TI - In brain metastasis systemic chemotherapy has been of no apparent value. PMID- 6282521 TI - "Hot spot" on perfusion lung scan produced by bronchiolo-alveolar cell carcinoma. PMID- 6282522 TI - Accumulation of Tc-99m phosphorus compounds in medullary carcinoma of the thyroid: report of two cases. PMID- 6282523 TI - Screening small for gestational age babies for congenital infection. AB - The widespread practice in newborn nurseries of screening asymptomatic small for gestational age (SGA) babies for TORCH infection has been evaluated. In a retrospective review, we found that, in 1979, in our nursery 23 (35%) of the sixty-six SGA babies were investigated for TORCH infection. No asymptomatic baby was investigated adequately to exclude infection. The two proven cases of congenital infection were both apparent on other clinical grounds, and neither would have been detected by our routine serologic screening. A review of published information on asymptomatic TORCH infections showed that, in the absence of other clinical signs of infection, intrauterine growth retardation is an unusual manifestation. Clinical investigation of TORCH infection should be confined to those babies with other clinical evidence of infection. PMID- 6282524 TI - Canrenoate reversal of inhibitory effects of digoxin on basal and furosemide stimulated renin secretion. AB - Changes in plasma renin activity (PRA) were monitored in six mildly hypertensive men after intravenous doses, in seven separate experiments, of placebo, digoxin, potassium canrenoate, potassium canrenoate with digoxin, furosemide, furosemide with digoxin, and potassium canrenoate with furosemide and digoxin. Potassium canrenoate has been used as a rapid source of canrenone, which has been recently shown to be a competitive antagonist of ouabain at its Na-K-ATPase receptor site. Potassium canrenoate infusion reversed the hyporeninemic effect of digoxin. This result has been taken as evidence that: (1) antialdosteronic drugs can also reverse digoxin effects at extracardiac level and (2) the Na-K-ATPase system is involved in the renin secretory mechanism. A seemingly identical reversal of the hyporeninemic effect of digoxin was induced by furosemide, which, when given alone, stimulated renin secretion. The simultaneous administration of potassium canrenoate, digoxin, and furosemide induced an increase in PRA on the same order as that after furosemide alone. This result indicates that furosemide stimulates renin release by affecting a biochemical system other than that affected by digoxin. PMID- 6282525 TI - Digoxin does not alter plasma steroid levels in health men. AB - Digoxin has been reported to induce feminizing effects in man. It does not compete for estradiol cytosol receptors in human breast carcinoma cells, however, and has no uterotrophic effect. We therefore investigated whether feminization might be due to digoxin action on plasma concentrations of sex steroids. Six healthy men (31.5 +/- 4 yr old) received therapeutic doses of digoxin for 43 days. We measured plasma concentrations of testosterone, androstenedione, dehydroepiandrosterone, estrone, estradiol, progesterone, 17-hydroxyprogesterone, cortisol, and aldosterone. During 35 days on digoxin levels of these steroids remained in the normal range and there was no change from before-drug values. Digoxin was in the therapeutic range of 1.9 +/- 3 nmol/l throughout. After stimulation by adrenocorticotropic hormone or human choriongonadotropin, the rise in plasma steroids was in the same range as when digoxin was given, as well as 16 wk after it had been discontinued. A normal rise in luteinizing hormone after luteinizing hormone-releasing hormone showed that the hypothalamogonadal feedback was not altered by digoxin. Free testosterone, estradiol, and cortisol concentrations under basal conditions and after stimulation were also the same before and after drug. It is concluded that the estrogen-like activity of digoxin cannot be explained by altered steroid availability from plasma. Feminizing effects attributed to digoxin may be caused by other conditions known to influence sex steroid hormones that are common in patients with heart disease. Our data suggest that digoxin may be the preferred digitalis therapy to avoid feminizing effects. PMID- 6282526 TI - Alpha-receptor function changes after the first dose of prazosin. AB - The effect of oral prazosin on blood pressure and antagonism of phenylephrine induce blood pressure increase was investigated in six healthy subjects during a dosing interval after the first dose and 3 days after the first dose of the drug. Prazosin lowered standing blood pressure more after the first dose than after the same dose 3 days later, despite similar plasma levels. Blood pressure decrease correlated with plasma prazosin levels during the elimination phase at the first dose, but not after 3 days of therapy. The phenylephrine log dose-response curves shifted to the right after prazosin, which indicates alpha-receptor antagonism of the drug. On day 4, the phenylephrine curve before prazosin dose shifted to the right of the pretreatment curve on day 1, despite very low prazosin plasma levels. On day 4 after after prazosin dosing the phenylephrine dose-response curves were shifted to the left of that on day 1. Our data indicated tolerance to prazosin effect on blood pressure and to phenylephrine agonism after 3 days dosing. Our data suggest that this might be due to desensitization of alpha adrenoceptors with differential effects of agonists and antagonists. PMID- 6282527 TI - Effects of enalapril, a new converting enzyme inhibitor, in hypertension. AB - The new angiotensin converting enzyme inhibitor enalapril maleate was given in single oral doses of 2.5, 5, and 10 mg to 11 hospitalized patients with uncomplicated essential hypertension who were on a 150-mEq sodium diet. All doses of enalapril induced reduction of mean seated diastolic blood pressure (SDBP). The magnitude of the initial SDBP reduction was not dose related, but the duration of effect was longer (greater than 12 hr) after the 5 and 10 mg. After dosing, mean plasma angiotensin converting enzyme activity (ACE) and aldosterone concentration (PAC) fell, while plasma renin activity (PRA) rose. Serum concentrations of the active diacid from of enalapril increased linearly with dosage; ACE was inhibited maximally at concentrations above 10 ng/ml. During repeated dosing in the outpatient trial there was attenuation of the antihypertensive effect (12 to 24 hr after dosing) in eight of 10 patients. Despite dose increases only two patients achieved SDBP control (less than or equal to 90 mm Hg). In the five patients in whom 50 mg/day hydrochlorothiazide was added near the end of the trail mean SDBP was further reduced. Enalapril was well tolerated. Further studies of the drug, especially in combination with diuretic, are needed. PMID- 6282528 TI - [Enkephalins and endorphins, peptide neurotransmitters regulating cerebral homeostasis: biological actions, biochemical properties, therapeutic prospects]. PMID- 6282529 TI - Phylogeny and ontogeny of the phosphoglycerate mutases--IV. Distribution of glycerate-2,3-P2 dependent and independent phosphoglycerate mutases in algae, fungi, plants and animals. AB - 1. The distribution of the glycerate-2,3-P2 dependent and independent phosphoglycerate mutases (PGM) has been studied in more than eighty species. 2. PGM activity in the extracts has been measured in the presence and in the absence of glycerate-2,3-P2, at pH 7.5 and at pH 8.5. 3. All samples with glycerate-2,3 P2 dependent PGM possess higher activity at pH 7.5 than at pH 8.5. In contrast, samples with glycerate-2,3-P2 independent PGM possess lower activity at pH 7.5 than at pH 8.5. 4. In algae and fungi both glycerate-2,3-P2 dependent and independent PGM have been found. 5. In plants only glycerate-2,3-P2 independent PGM has been detected. 6. In animals both types of PGM are present. Independent PGM activity is present in sponges, coelenterates, myriapods, arachnids and echinoderms. Glycerate-2,3-P2 dependent PGM is present in platyhelminths, mollusks, annelids, crustaceans, insects and vertebrates. PMID- 6282530 TI - Diabetic nutrition: come back, carbohydrates! PMID- 6282531 TI - Deaths following female sterilization with unipolar electrocoagulating devices. Centers for Disease Control H.E.W., Public Health Service, Atlanta, GA. PMID- 6282532 TI - Bovid herpesvirus 2: natural spread among breeding bulls. AB - Sera collected over a seven year period, together with detailed health and management records, were used in a retrospective study of Bovid herpesvirus 2 (BHV-2) spread within a large herd of breeding bulls. Virus spread rapidly throughout the bulls in one barn. Transfer of infected bulls to other barns was followed by further, although sporadic, spread of virus. Spread of BHV-2 from seropositive animals to susceptible bulls in close contact led to the conclusion that reactivation and transmission of latent virus had occurred. Semen from seropositive bulls did not transmit BHV-2 to seronegative calves that were purposefully inoculated by the intranasal or intravenous routes. PMID- 6282533 TI - Compartmentation of newly synthesized proteins. PMID- 6282534 TI - The main types of organization of genetic material in eukaryotes. PMID- 6282536 TI - Endogenous retroviruses of mice and chickens. PMID- 6282535 TI - Affinity labeling via deamination reactions. AB - An electrophilic center at saturated carbon generated by the departure of molecular nitrogen shows minimum discrimination between various nucleophiles. The generation of such a center in the active site of a protein is therefore an attractive way of labeling that active site. The chemistry of deamination reactions will be discussed with respect to the practicality of triggering the deamination in the active sites of proteins. Successful applications of this principle using the N-nitrosamide functionality, the alkyl aryl triazene functionality, and the diazo functionality will be described. Reasons why active site reagents incorporating this type of covert electrophilicity are more specific than those incorporating an overtly electrophilic center (such as -CO CH2-Halogen) will be advanced. The actual and potential application of deamination precursors to the specific inhibition of physiological activities in living cells will be discussed. PMID- 6282537 TI - Expression of xenotropic murine leukemia viruses. PMID- 6282538 TI - Possible immunological mechanisms in C-type viral leukemogenesis in mice. PMID- 6282539 TI - Steroid acne in a 14-month-old boy. PMID- 6282540 TI - A case in which the association between basal cell epithelioma and histiocytoma is demonstrated. PMID- 6282541 TI - A model for service delivery research and evaluation: management implications for alcohol, drug abuse and mental health organizations. AB - Public alcohol, drug and mental health organizations have failed to adequately demonstrate their impact. Historically, the justification for these programs have relied more on good intentions and good faith to support their efforts than on their documented efficacy. This lack of documentation has contributed, in part, to recent federal and state mandates for better management, control, and evaluation of publicly financed mental health, alcohol and drug abuse care. This paper presents a model of applied research and evaluation designed to enhance the capabilities of public mental health, drug and alcohol programs in demonstrating their clinical and rehabilitative results. PMID- 6282542 TI - Effects of disulfiram on the sleep of chronic alcoholics. AB - Thirty male alcoholic inpatients received in a random double-blind design placebo (P) or disulfiram 250 mg. (D) at bedtime in order to evaluate the effects of the latter on EEG-EOG sleep patterns. Subjects (S's) had no major medical illness and required no medication for 3 weeks prior to the study. There were 13 disulfiram S's and 17 placebo S's. The total REM time decreased significantly (p less than 0.05) in the D group. Total number of REM episodes decreased and latency to stage 1 REM increased in the disulfiram group. These results are characteristic of sleep patterns induced by the administration of catecholaminergics and opposite to those induced by catecholamine inhibiting agents. PMID- 6282543 TI - Alcoholism programs in the USSR and Yugoslavia: effects of the social context on treatment. AB - The social context in which alcoholism treatment takes place contributes greatly to its nature and outcome. This issue is examined in two socialist countries, based on site visits and literature review. A hospital-based treatment program and its related social Club system in Yugoslavia are part of a close-knit national treatment network oriented toward self-help and peer therapy. The principal approaches to alcoholism treatment in the USSR are closely integrated into the Communist social and political structure, and include sobering-up stations, factory-based clinics, hospital-based inpatient and outpatient facilities, and a quasi-penal system for recidivists. The impact of the following issues on alcoholism treatment is discussed: (1) the public information system, in both ideology and propaganda; (2) the state's regulation of deviant behavior, as a component of the medical treatment system; and finally, (3) the acceptable role for peer treatment in a given social context. PMID- 6282544 TI - Membrane fluidity and alcohol actions. AB - Many studies have demonstrated that the ability of alcohols and other intoxicant anesthetics to affect biochemical, physiological, and behavioral processes rests in their hydrophobicity. This means that potency is determined by the ability of the drug to move from a water phase into a lipid or membrane phase. In more precise terms, anesthetic effects are correlated with the volume occupied by the anesthetic molecules within the membrane. Although the anesthetic effects, particularly the inhibition of nerve condition, have been used most frequently in establishing this correlation, the intoxicating effects (i.e., ataxia) of a series of alcohols had also been correlated with their membrane partitioning. These results suggest that the intoxicating, as well as anesthetic, effects of ethanol and related drugs are due to their penetrating into hydrophobic regions of nerve membranes. The predominant hydrophobic region of biological membranes is the "sea" of lipid that surrounds "islands" of functional proteins. This leads to the postulate that intoxicant-anesthetics alter the physical properties of membrane lipids and thus affect neuronal function. To evaluate this hypothesis, we must consider the lipid composition of brain membranes, the importance of membrane lipids in neuronal function, the techniques available for the study of membrane physical properties, and the effects of ethanol on nerve membranes. PMID- 6282545 TI - Fiberoptic bronchoscopic laser photoradiation for tumor localization in lung cancer. AB - Hematoporphyrin derivative (HpD) administration and laser photoradiation were combined to clarify the system's clinical applicability in localizing lung cancer tumors. It was used in 16 cases of lung cancer and in one case of severely atypical squamous metaplasia. Tumors were irradiated 48 hours after intravenous injection of 2.0 to 4.0 mg/kg HpD and fluorescence observed. Fluorescence was seen in 13 lung cancer cases in the case of squamous metaplasia, but results were negative in three of the cases of lung cancer. Among the positive cases was one of occult cancer and two invading the trachea, difficult to recognize by conventional fiberoptic bronchoscopy, as was the severely atypical squamous metaplasia. In the three lung cancer cases not fluorescing, the tumors were obscured by blood, necrotic tissue, or normal mucosa. The method holds promise in the diagnosis of malignant tumors. Nevertheless, it needs further refinement and more studies to elucidate definitive differentiation between malignant and severely atypical, nonmalignant tissue. PMID- 6282546 TI - Patholphysiology of asthmatic bronchoconstriction. PMID- 6282547 TI - Multicentric malignant fibrous histiocytomas of the colon. Report of a case and review of the subject. AB - Malignant fibrous histiocytoma is a primitive mesenchymal tumor usually found in the soft tissues of the extremity or torso of adults. Only two previous reports of this lesion in the abdominal viscera are present in the world medical literature. A case is reported of three synchronous tumors in the rectum, colon, and omentum with the pathologic appearance of malignant fibrous histiocytoma. Although resected completely with a transverse colocolostomy and a low-anterior resection, the expected behavior of this tumor is one of aggressive local recurrence followed by distant metastasis. The clinical and pathologic characteristics of malignant fibrous histiocytoma are reviewed. PMID- 6282548 TI - Pancreatic enzyme replacement: the effect of antacids or cimetidine. AB - The effect of the addition of sodium bicarbonate, aluminum hydroxide, magnesium hydroxide, calcium carbonate, or cimetidine to supplemental pancreatic enzyme therapy was investigated in patients with severe exocrine pancreatic insufficiency. Steatorrhea was reduced with the administration of three enzyme tablets with meals (73 vs 29 g/24 hr). The coadministration of enzyme tablets with either sodium bicarbonate (16.6 g/24 hr, P = 0.08), or aluminum hydroxide (18.4 g/24 hr, P = 0.029) yielded a greater reduction in steatorrhea than enzymes alone (29 g/24 hr). Neither magnesium-aluminum hydroxide (36.3 g/24 hr, P = 0.22), nor calcium carbonate (39.0 g/24 hr, P = 0.029), improved efficacy of enzyme therapy and, in fact, tended to enhance steatorrhea. With the administration of cimetidine there was no significant effect on steatorrhea compared to enzymes alone (32.1 vs 29 g/24 hr, P greater than 0.3). Intraduodenal lipase activity following test meals was found to be a poor predictor of the effectiveness of antacid therapy in improving the efficacy of supplemental enzymes. PMID- 6282549 TI - Semipurified dietary fiber and small-bowel morphology in rats. AB - Newly weaned rats fed 12 weeks on a diet containing no dietary fiber or no fiber except for 10% cellulose, maintained the leaf-like intestinal villous morphology present at weaning, as observed by scanning electron microscopy. In rats on a normal laboratory diet the jejunal morphology showed progression from the leaf like villous pattern at weaning to broad-leafed, long-ridged villi of adulthood. Pectin added to a no-fiber diet caused structural changes similar to but less well developed than those changes in the rats on a standard diet. Striking differences were noted not only in the appearance of the intestinal villi but in the number of villi per square centimeter between those animals on no fiber or no fiber except cellulose and those animals on pectin or standard diets. Cholestyramine, a strong pharmacological bile salt-binding agent, when added to a no-fiber diet, did not promote development of the usual villous pattern, and the structure remained the same as that in rats on no-fiber and cellulose diets. Cellulose (no bile salt-binding capability) and pectin (weak bile salt-binding capability) added to a no-fiber diet were associated with significant differences in the number of villi in both the jejunum and the ileum. The observed changes in morphology are unlikely to be due to differing bile salt-binding capabilities of different fiber substances. PMID- 6282550 TI - Effects of pH on in vitro gallbladder responses to cholecystokinin octapeptide. AB - We investigated the effect of pH change on octapeptide cholecystokinin's ability to contract guinea pig gallbladder strips. In the absence of exogenous drugs, pH changes had no demonstrable effect on gallbladder tension. However, when gallbladder strips were contracted with CCK-OP at pH 7.3, increasing the pH in the muscle bath to 7.8 produced further increases in tension at each dose studied (P less than 0.025). Subsequently, decreasing the bath pH to 6.9 decreased the strip tension to less than that observed at pH 7.3 (P less than 0.025). Reversing the order in which these pH alterations were made produced similar results; a pH decrease from 7.3 to 6.9 decreased the response to CCK-OP at the two highest doses (P less than 0.025), while subsequent elevation of pH raised the tension to levels greater than those observed at pH 7.3 (P less than 0.01). Furthermore, contracting the strips at pH 8.0 and slowly decreasing pH at a rate of 0.1 pH units per minute consistently produced an acceleration of tension decay as pH fell. After contracting the strip at pH 6.7, slow increases in pH reversed the tension decay and enhanced the contractile response as pH was increased from 6.7 to 8.0. These results demonstrate that pH changes alter the gallbladder contractile response to cholecystokinin. Although this phenomenon has been reported for a number of other physiologic agents, we believe this is the first such demonstration for a peptide hormone. PMID- 6282551 TI - Verapamil: a calcium-channel blocker. PMID- 6282552 TI - Herpetic whitlow. PMID- 6282553 TI - The postenzymatic chemistry of activated oxygen. PMID- 6282554 TI - [Cushing's syndrome (author's transl)]. PMID- 6282555 TI - [Treatment of Wilms' tumor (author's transl)]. AB - Uniform treatment based on the therapeutic approach of the 1st and 2nd US National Wilms' Tumor Study was decided on in March 1976 by paediatricians, surgeons, urologists and radiotherapists in Austria. Wilms' tumour was diagnosed in 34 children between 1 january 1976 an 29 february 1980 (stage I: n = 11, stage II: n = 8, stage III: n = 8, stage IV: n = 7). Parents of two children refused treatments; both children have since died of metastases. Of the remaining 32 children 29 (90.6%) are alive, 10 for more than 4, 15 for more than 3 and 19 for more than 2 years after diagnosis. 21 children are without need of treatment. Three children have died, one due to postoperative complications, one due to haemorrhagic chickenpox, but free of tumour, and one after insufficient treatment. Two of the five children with a recurrence between 2 1/4 to 15 months after diagnosis had been treated inadequately in the initial phase. The tumour free survival rate in 74.2%. Two children with early occurring or recurrent lung metastases have survived for 53 1/2 and 54 months up to now. PMID- 6282556 TI - [Drug therapy of pituitary tumors]. PMID- 6282557 TI - [Programmed after care of breast cancer. Results of a follow-up model after 2 years later]. AB - In two years 359 patients with breast cancer were evaluated in a cooperative follow-up programme in which the organisation and documentation were done in hospital, the medical care and investigations by the family doctor. 72% of 316 evaluable patients remained in regular control. 63% chose the family doctor for follow-up while the others preferred hospital. In the first year 71-80% attended their appointments, in the second year 60-74%. Statistically significant parameters for detection of metastases were the following: general condition, local signs, palpable lymph nodes, markedly increased ESR, alkaline phosphatase, chest radiograph, and bone scan. Cumulative freedom from recurrence in the whole group diminished from 1.0 to 0.631 in 25 months, cumulative survival from 1.0 to 0.876 in 20 months. In both groups there were significant differences between stages I, II and III. Forty patients (16%) out of 250 developed a recurrence within 2.25 years, in five (2%) a second carcinoma developed. There was a high proportion of local recurrences (20 out of 40) and these often led to generalised disease (11 out of 20). PMID- 6282558 TI - [Verdict of the Supreme Court of the Federal Republic of Germany on advertising by non-medical practitioners]. PMID- 6282559 TI - [Antidiuretic hormone in patients with bronchial carcinoma (author's transl)]. AB - Antidiuretic hormone (ADH) was determined by radioimmunoassay in 139 patients with bronchial carcinoma. Serum ADH levels, compared with a control group, were increased in 30% of patients with small-cell and 21% of patients with large-cell bronchial carcinoma. Patients with squamous carcinoma or adenocarcinoma had normal ADH values. There was no correlation between serum ADH levels and stage of tumour. Serial ADH measurements during chemotherapy provided good correlation between ADH and response to treatment. ADH was also demonstrable by immunohistology in the tumour cells of one patient with increased serum levels. PMID- 6282560 TI - [Low-symptom herpes simplex encephalitis (author's transl)]. AB - A 39-year-old man had acoustic hallucinations for about one year due to chronic alcoholism. He suddenly lost consciousness after suffering from headaches for about four weeks. EEG and CT scan were indicative of a brain tumour but lumbar puncture revealed slight pleocytosis and an immunoglobulin G fraction synthesized within the central nervous system. This secretory fraction contained major portions of locally produced herpes simplex antibodies detectable with a complement fixation test and a recently developed enzyme immunoassay. The patient remained in hospital for only a few days and went back to work after six weeks. To our knowledge this is the first case reported surviving herpes simplex encephalitis outside hospital without impairment of consciousness, confusion or substantial neurologic symptoms. The question remains open, whether the generally severe course of this disease had been modified by the chronic alcoholism. It is obvious that the correct diagnosis would have been missed without lumbar puncture. PMID- 6282561 TI - [Opioid peptides and their receptors]. PMID- 6282562 TI - [Hereditary pressure-sensitive neuropathy]. PMID- 6282563 TI - Multiple primary malignant neoplasms a report of 2 Nigerian patients with double synchronous malignancies. PMID- 6282564 TI - Single fibre EMG for the investigation of extraocular eye muscles. PMID- 6282565 TI - Mechanisms by which calcium ions regulate the steroidogenic actions of luteinizing hormone in isolated ovarian cells in vitro. AB - Incubation of swine granulosa cells in chemically defined medium selectively deficient in calcium ions markedly impaired progesterone production in response to submaximal and maximally stimulating concentrations of LH. Accumulation of progesterone in response to LH was reduced significantly in both cells and medium, without a discernible shift in the time-course of progestin production. The reduction in progesterone accumulation could not be accounted by increased formation of the catabolite 20 alpha-hydroxypregn-4-en-3-one. In addition, progesterone secretion basally or in response to exogenously supplied pregnenolone was not altered in calcium-deficient incubations. Administration of verapamil or diltiazem, organic inhibitors of net transmembrane calcium uptake, also suppressed LH-stimulated progesterone production. Conversely, micromolar concentrations of the divalent cation ionophore A23187 significantly enhanced the stimulatory effects of LH. The mechanisms of calcium action were examined further in relation to the cAMP effector system. Calcium deprivation significantly suppressed the dose-dependent accumulation of cAMP in granulosa cells treated with LH but had no effect on basal levels. Omission of calcium ions from the extracellular medium also markedly impaired production of progesterone in response to 8-bromo-cAMP, cholera toxin, or 3-isobutyl-l-methylxanthine. The present studies suggest that calcium ions significantly modulate LH-stimulated progesterone biosynthesis in isolated ovarian cells in vitro. Specific regulatory actions of calcium ions in granulosa cells may be exerted at several levels, including LH-stimulated cAMP accumulation and at intracellular loci distal to the actual generation of cAMP. PMID- 6282566 TI - Gonadotropin-releasing hormone modulation of its own pituitary receptors: evidence for biphasic regulation. PMID- 6282567 TI - Activity ratio measurements reflect intracellular activation of adenosine 3',5' monophosphate-dependent protein kinase in osteoblasts. AB - Parathyroid hormone, prostaglandin E2, and prostacyclin activate cAMP-dependent protein kinase in osteoblast-rich normal rat calvarial cells and in clonal rat osteogenic sarcoma cells of osteoblastic phenotype. The present study was undertaken to determine the activation of the enzyme in relation to cellular cAMP concentrations at increasing doses of the three hormones and also to test that the activity ratio measurement of the enzyme (ratio of the activity in the absence of cAMP to the activity in the presence of excess cAMP) was a true reflection of intracellular activation of the enzyme. With each hormone, using either normal or malignant osteoblasts, activation of the enzyme took place at hormone concentrations lower than those required to produce detectable changes in cAMP concentrations in the incubations. Stimulation of activity was abolished by addition of the heat-stable inhibitor of cAMP-dependent protein kinase, indicating that activation was of cAMP-dependent protein kinase alone. To demonstrate that protein kinase activation occurred intracellularly and not during sample preparation, charcoal was added at the time of cell disruption to absorb free cAMP. Under these conditions, no change was observed in the concentration of bovine parathyroid hormone required to cause activation of cAMP dependent protein kinase. Finally, addition of purified cAMP-dependent protein kinase type I or type II to treated cells at the time of lysis did not result in significant activation of added isoenzyme, except at hormone concentrations sufficient to increase the total cAMP concentration of incubations. It is concluded that activity ratio measurement reflects the intracellular state of activation of cAMP-dependent protein kinase in the osteoblast-like cells treated by hormones and, furthermore, that only a fraction of the maximally generated cAMP is necessary for full enzyme activation. PMID- 6282568 TI - Metoclopramide inhibits aldosterone biosynthesis in vitro. AB - Metoclopramide, a dopaminergic antagonist, has consistently elevated plasma aldosterone levels in vivo. To determine whether this was a direct action of metoclopramide on adrenal steroidogenesis, we examined the response of collagenase-dispersed rat adrenal glomerulosa cells to metoclopramide in vitro. The effect of increasing concentrations of metoclopramide (3 X 10(-10) to 3 X 10( 4) M) on basal as well as angiotensin II (2.4 X 10(-10) to 2.4 X 10(-8) M)-, ACTH (3.5 X 10(-11) M)- and potassium (5.9 meq/liter)-stimulated aldosterone production was evaluated. Metoclopramide caused a dose-related decrease in basal and stimulated aldosterone production (P less than 0.01). In addition, metoclopramide also blocked basal and stimulated corticosterone production (P less than 0.01). This was not due to an irreversible toxic effect, since glomerulosa cells preincubated with 3 X 10(-4) M metoclopramide excluded trypan blue dye and responded to ACTH stimulation. Sodium metabisulfite, an antioxidant present in the metoclopramide preparation, did not contribute to the metoclopramide effect. These results indicate that metoclopramide is an aldosterone antagonist in vitro, contrary to reported data obtained in vivo. Thus, metoclopramide may be a partial dopaminergic agonist: in vitro where dopamine levels are negligible, it is an agonist, whereas in vivo where dopamine concentrations are greater, it is an antagonist. PMID- 6282569 TI - Localization of delta 4-5 beta- and 5 alpha-reductases and 17 beta-ol dehydrogenase in immature golden hamster testes. AB - Testes of 28- and 70-day-old golden hamsters were separated into seminiferus tubules and interstitial tissue by wet dissection. Homogenates of the tissues and subcellular fractions were incubated with [14C]androstenedione, and NADPH and enzyme activity (nanomoles per 100 mg protein/h) was estimated. In immature testes, the 5 beta-reductase activity was largely confined to the seminiferous tubules, while 5 alpha-reductase was localized in the interstitial tissue. The activity of 17 beta-ol-dehydrogenase was found to be equally distributed between tubules and interstitial tissue. Although the activities of 5 beta- and 5 alpha reductases were approximately 10 times greater in immature than adult testes, the 17 beta-ol-dehydrogenase activity in adult testes was approximately 10 times that in immature testes. In the immature testes, most 5 alpha-reductase and 17 beta-ol dehydrogenase were localized in microsomes, whereas 5 beta-reductase was wholly localized in cytosol. These results indicates that 5 beta-reduction of delta 4-3 ketosteroids takes place largely in cytosol of tubular cells, while 5 alpha reduction occurs in microsomes of interstitial cells in the testis of the immature golden hamster. PMID- 6282570 TI - I alpha, 25-Dihydroxyvitamin D3 receptor in the X-linked hypophosphatemic mouse. AB - We have attempted to determine if the resistance of the X-linked hypophosphatemic mouse to the actions of 1,25(OH)2D3 is due to abnormal cytosolic receptors for this hormone. Cytoplasmic 1,25-dihydroxy-vitamin D3 [1,25(OH)3D3] receptors were demonstrated in hypophosphatemic (HYP) mouse tissues including intestine, kidney and testis. Cytosol preparations from 14 murine tissues were prepared using hypertonic buffer and incubated for three hours at 0 C with 1,25(OH)2 [23,24(n)-( 3)H]D3 ([3H]-1,25(OH)2D3). The results of studies using 5-20% sucrose density gradients revealed th at cytosol preparations from intestine, kidney and testis exhibited a 3.2 S peak which ws specific for 1,25(OH)2D3. Scatchard analysis of intestinal, renal and testicular cytosol preparations of Hyp mice revealed a single class of noninteracting binding sites with a range of equilibrium dissociation constants (KD) of 0.9-3.5 X 10(-10) M, and a range of specific binding sites of 15-317 fmol/mg protein. There were no significant differences in values of KD, and numbers of specific binding sites among Hyp and control mice. We conclude the reported resistance of Hyp mice to vitamin D is not due to abnormal 1,24(OH)2D3 receptor in Hyp mouse. PMID- 6282571 TI - Potency enhancement of a GnRH agonist: GnRH-receptor microaggregation stimulates gonadotropin release. AB - Binding of gonadotropin releasing hormone (GnRH, pyro-Glu1-His2-Trp3-Ser4-Tyr5 Gly6-Leu7-Arg8-Pro9-Gly-NH210) to its plasma membrane receptor is the first step leading to pituitary luteinizing hormone (LH) release. In the present study, we have prepared the ethylene glycol bis(succinimidyl succinate) (EGS) dimer of a GnRH agonist, D-Lys6-GnRH; that is, (D-Lys6-GnRH)-EGS-(D-Lys6-GnRH). The bridge length of the EGS is about 15A. This dimer stimulates LH release from pituitary cultures with full efficacy but slightly less potency than D-Lys6-GnRH indicating that the dimerization merely restricts the action of each molecule of D-Lys6-GnRH with respect to the other. When a concentration of the dimer, which alone is too low to evoke substantial LH release, is incubated with pituitary cell cultures in the presence of antibody (AB) against D-Lys6-GnRH, considerable potency enhancement occurs. LH release in response to the AB-dimer conjugate requires extracellular Ca2+ and is blocked by Pimozide. The monovalent form (i.e. reduced pepsin fragment) of AB is ineffective in stimulating release. The addition of antibody to the dimer appears to confer the ability to cross-link receptors and indicates that receptor microaggregation as such is sufficient to activate the effector system in these cells and evoke release. PMID- 6282572 TI - Pituitary LHRH receptors during puberty in the female rat: is the proestrous decline in receptor content due to occupancy? AB - As during the adult estrous cycle, the number of pituitary luteinizing hormone releasing hormone (LHRH) receptors was found to increase prior to the first preovulatory surge of LH. Receptor content, as measured by the binding of the analog 125I-D-Ala6-Pro9-LHRH to pituitary membranes, declined markedly at the time of the first LH surge, remaining low during the first estrus and first diestrus. When pituitary membranes from animals undergoing an LH surge were incubated with MgCl2 to dissociate endogenously bound hormone, available binding sites were restored to pre-LH surge values. The increase in LHRH binding capacity was not related to an increase in receptor affinity, and it was not demonstrable in other pubertal phases. MgCl2 may have unmasked receptors made cryptic at the time of the LHRH-induced LH surge. Nevertheless, the results are compatible with the view that the proestrous decline in LHRH receptor content is, at least in part, due to occupancy of binding sites by endogenous hormone. PMID- 6282573 TI - Beta-adrenergic stimulation of prolactin release from superfused pituitary cell aggregates. AB - l-Isoproterenol (l-ISO), a specific agonist of beta-adrenergic receptors, evoked a prompt rise of prolactin (PRL) release from superfused anterior pituitary cell aggregates established in culture for 5 days. The effect was concentration dependent between 1 and 100 nM. d-Isoproterenol was more than 2 orders of magnitude weaker than the l-isomer. When dopamine receptors were blocked with domperidone, PRL secretion was also stimulated by l-epinephrine (E) and l norepinephrine (NE), the rank order of potency being l-ISO greater than E much greater than NE. Under the latter conditions dopamine and the alpha-adrenergic agonists, clonidine and phenylephrine, had no stimulatory effect at 1 microM. Stimulation of PRL release by l-ISO and E was blocked by the beta-receptor antagonist, propranolol, but not by the alpha-receptor blocker, prazosin. PMID- 6282574 TI - In vivo evidence for increased adrenal sensitivity to adrenocorticotropin-(1-24) in the lamb fetus late in gestation. AB - Plasma concentrations of immunoreactive ACTH and cortisol were monitored daily in chronically cannulated sheep fetuses during the last 3 weeks of gestation. A prepartum increase in fetal plasma cortisol occurred without a concomitant rise in fetal plasma ACTH. When fetal lambs were injected with various doses of ACTH (1-24) and the plasma cortisol responses were integrated over time, the resulting changes in the log dose-response curves indicate that fetal adrenal sensitivity increases late in gestation. Thus, the marked rise in fetal plasma cortisol before birth can be explained, at least in part, by an increase in fetal adrenal sensitivity to ACTH. PMID- 6282575 TI - Effects of sodium repletion and timolol maleate administration on the zona glomerulosa of the rat adrenal cortex: an electron microscopic morphometric study. AB - The effects of long-term sodium repletion and timolol maleate treatment on the zona glomerulosa of dexamethasone- and ACTH-administered rats were investigated. Morphometry showed that both treatments induced a noticeable atrophy of the zona glomerulosa and its parenchymal cells, which is mainly due to the decrease in the smooth endoplasmic reticulum and the mitochondrial compartment. Radio immunological studies indicated that in the treated rats the plasma aldosterone concentration was significantly decreased. Combined sodium repletion and timolol maleate administration provoked more intense effects than the single treatments. These data confirm the contention that both Na/K balance and the renin angiotensin system are involved in the maintenance of the growth and steroidogenic capacity of the rat zona glomerulosa, with interfering without the hypothalamo-hypophyseal-adrenal axis. PMID- 6282576 TI - Corticotropin-releasing factor. PMID- 6282578 TI - Translocation of asbestos dust through the bronchiolar wall. PMID- 6282579 TI - Influence of the sialic acid content of the membrane on its susceptibility to chrysotile. PMID- 6282577 TI - Toxicity of polybrominated biphenyls (PBBs) in Domestic and laboratory animals. AB - The composition, environmental fate, and effects of the polybrominated biphenyls (Firemaster BP-6 or FF-1) involved in the accidental contamination of cattle feed in Michigan in 1973 are reviewed. Toxic effects referred to in this report are limited to those occurring in domestic and laboratory animals and include general toxicity, neurobehavioral toxicity, immunotoxicity, reproductive toxicity, mutagenicity and carcinogenicity. The absorption, distribution, biotransformation and elimination of these polybrominated biphenyls are discussed along with the interactions with other chemicals and drugs. PMID- 6282580 TI - Absence of mutagenic activity of three forms of asbestos in liver epithelial cells. PMID- 6282581 TI - Immunological alterations in the mouse following injection or inhalation of silica and olivine dusts. PMID- 6282582 TI - Carcinogenic and fibrogenic effects of zeolites: preliminary observations. PMID- 6282583 TI - Haematological changes in 2 ponies before and during an infection with equine influenza. PMID- 6282584 TI - Two identical subunits of the EcoRI restriction endonuclease Co-operate in the binding and cleavage of the palindromic substrate. AB - The cleavage of radioactively labelled double-stranded d(G-G-A-A-T-T-C-C) was studied in single turnover experiments with substrate and enzyme both being in the micromolar range. The reaction rate was found to increase with enzyme concentration until a ratio of one tetrameric enzyme to two double-stranded substrates was reached, further increase of the enzyme concentration then leads to a sharp decline of the reaction rate. These findings are interpreted in the following manner. (a) Two subunits of the EcoRI endonuclease co-operate in binding and possibly also in cleaving the palindromic substrate. (b) The enzymatic action of the EcoRI endonuclease is inhibited by excess enzyme, possibly due to unspecific binding of the enzyme-substrate complex. The self inhibition of EcoRI endonuclease has also been observed with macromolecular substrates. PMID- 6282585 TI - Fructose-2,6-bisphosphatase from rat liver. AB - An enzyme that catalyzes the stoichiometric conversion of fructose 2,6 bisphosphate into fructose 6-phosphate and inorganic phosphate has been purified from rat liver. This fructose 2,6-bisphosphatase copurified with phosphofructokinase 2 (ATP: D-fructose 6-phosphate 2-phosphotransferase) in the several separation procedures used. The enzyme was active in the absence of Mg2+ and was stimulated by triphosphonucleotides in the presence of Mg2+ and also by glycerol 3-phosphate, glycerol 2-phosphate and dihydroxyacetone phosphate. It was strongly inhibited by fructose 6-phosphate at physiological concentrations and this inhibition was partially relieved by glycerol phosphate and dihydroxyacetone phosphate. The activity of fructose 2,6-bisphosphatase was increased severalfold upon incubation in the presence of cyclic-AMP-dependent protein kinase and cyclic AMP. The activation resulted from an increase in V (rate at infinite concentration of substrate) and from a greater sensitivity to the stimulatory action of ATP and of glycerol phosphate at neutral pH. The activity of fructose 2,6-bisphosphatase could also be measured in crude liver preparations and in extracts of hepatocytes. It was then increased severalfold by treatment of the cells with glucagon, when measured in the presence of triphosphonucleotides. PMID- 6282586 TI - Influence of the structure of water on the hydrolysis of cytidine 2',3'-phosphate catalysed by bovine pancreatic ribonuclease A. AB - The study of the temperature dependence of the hydrolysis of cytidine 2',3' phosphate by bovine pancreatic ribonuclease A (EC 3.1.27.5) at pH 7.0 by using the pH-stat method showed a transition at 4 degrees C [J. A. Biosca and C. M. Cuchillo (1980) Biochem. J. 189, 655-657]. The breaks found in the Van't Hoff and Arrhenius plots at pH 7.0 were confirmed in the present work by following the reaction spectrophotometrically with a stopped-flow spectrophotometer adapted to the use of sub-zero temperatures. Similar results were found at pH 5.5. In addition it was found that the discontinuity disappears when 40% ethyleneglycol is present in the reaction mixture. However, in this latter instance a discontinuity around 0 degrees C appears in the Arrhenius plot. To explore the possibility that all these effects were due to a conformational transition in the protein, thermal perturbation experiments were carried out with the enzyme. A change in the slope of the plot of delta A290 as a function of temperature was found around 6 degrees C at pH 7.0 but not at pH 5.5. The results reported here can be interpreted as due to a change in the protein structure induced by the change of the structure of water. The studies carried out in the presence of ethyleneglycol also open the way to the cryoenzymological experimentation on ribonuclease. PMID- 6282587 TI - Dimerization of deoxyribonuclease I, lysozyme and papain. Effects of ionic strength on enzymic activity. AB - Transition of bovine ribonuclease A from its monomeric to a dimeric form changes the pattern of enzymic activity response to ionic strength [Sorrentino, S., Carsana, A., Furia, A., Doskocil, J., and Libonati, M. (1980) Biochim. Biophys. Acta. 609, 40-52]. To see whether this phenomenon could be common to other enzyme substrate systems, the action of various dimeric and monomeric enzymes (ox pancreas deoxyribonuclease, hog spleen acid deoxyribonuclease, bovine seminal ribonuclease, egg-white lysozyme, and papain) on polyelectrolytic substrates has been studied under different conditions of ionic strength. Dimerization of ox pancreas deoxyribonuclease, lysozyme and papain was obtained by cross-linkage with dimethyl suberimidate. The main results of the investigation, similar to those obtained with ribonuclease A, are the following. 1. Enzyme monomers and dimers show markedly different patterns of activity response to ionic strength at given pH values: the reactions catalyzed by monomeric enzymes are highly modulated by salt, whereas those catalyzed by dimeric enzymes are not. In particular, at the reaction optimum the monomeric form of an enzyme is significantly more active than the dimeric one. 2. The optimum of the reaction catalyzed by a dimeric enzyme is shifted to higher ionic strengths in comparison with that of the reaction catalyzed by a monomeric enzyme. A model is proposed that could explain these results on the basis of the influence of ionic strength on the intramolecular dynamics of the enzyme molecule and its non-specific interactions with polyelectrolytic substrates. PMID- 6282588 TI - Na+ channels with binding sites of high and low affinity for tetrodotoxin in different excitable and non-excitable cells. AB - The properties of interaction of tetrodotoxin with its receptor site on the voltage-sensitive Na+ channel were analysed in two ways: (a) by titrating Na+ channels with a tetrodotoxin derivative, [3H]ethylenediamine-tetrodotoxin; (b) by studying the physiological properties of interaction of the toxin with its receptor from 22Na flux measurements. Using a variety of cell types in culture, three different kinds of situations were observed. 1. Cells like N1E 115 neuroblastoma, CCl 39 fibroblasts, embryonic chick cardiomyocytes and chick skeletal myotubes only have one family of Na+ channels with high-affinity binding sites (in the nanomolar range) for tetrodotoxin. These Na+ channels are the same ones as those that are activated by the alkaloid and polypeptide toxins that accelerate 22Na+ influx. 2. C9 cells have Na+ channels with low-affinity binding sites for tetrodotoxin. These Na+ channels are also the ones that are activated by alkaloid and polypeptide toxins (the median inhibitory concentration for tetrodotoxin inhibition of 22Na+ influx through these Na+ channels in 300 nM). 3. Rat myotubes that have differentiated in culture in the absence of neuronal influence have both high-affinity binding sites (in the nanomolar range) detected with the tritiated tetrodotoxin derivative and low-affinity binding sites (on the micromolar range) detected by 22Na+ flux experiments. Only low-affinity binding sites correspond to Na+ channels that can be activated with alkaloid and polypeptide toxins. PMID- 6282589 TI - Separation and characterisation of glycogen synthase kinase 3, glycogen synthase kinase 4 and glycogen synthase kinase 5 from rabbit skeletal muscle. PMID- 6282590 TI - Limited proteolysis and proton NMR spectroscopy of Bacillus stearothermophilus pyruvate dehydrogenase multienzyme complex. AB - The pyruvate dehydrogenase multienzyme complex of Bacillus stearothermophilus was treated with chymotrypsin at pH 7 and 0 degrees C. Loss of the overall catalytic activity lagged behind the rapid cleavage of the lipoate acetyltransferase polypeptide chains, whose apparent Mr fell from 57 000 to 45 000 as judged by sodium dodecylsulphate/polyacrylamide gel electrophoresis. The inactive chymotrypsin-treated enzyme had lost the lipoic-acid-containing regions of the lipoate acetyltransferase chains, yet remained a highly assembled structure. Treatment of this chymotryptic core complex with trypsin at pH 7.0 and 0 degrees C caused a further shortening of the lipoate acetyltransferase polypeptide chains to an apparent Mr of 28 000 and was accompanied by disassembly of the complex. The lipoic-acid-containing regions are therefore likely to be physically exposed in the intact complex, protruding from the structural core formed by the lipoate acetyltransferase component between the subunits of the other component enzymes. Proton nuclear magnetic resonance spectroscopy demonstrated that the enzyme complex contains large regions of polypeptide chain with remarkable intramolecular mobility, most of which were retained after excision of the lipoic acid-containing regions with chymotrypsin. It is likely that the highly mobile regions are in the lipoate acetyltransferase component and facilitate movement of the lipoic acid residues. Such polypeptide chain mobility provides the molecular basis of a novel system of active-site coupling in the 2-oxo acid dehydrogenase multienzyme complexes. PMID- 6282591 TI - Spin-label characterisation of the lamellar-to-hexagonal (HII) phase transition in egg phosphatidylethanolamine aqueous dispersions. AB - The thermotropic behaviour of egg yolk phosphatidylethanolamine dispersions in excess aqueous phase has been investigated by spin label electron spin resonance spectroscopy and differential thermal analysis. Phosphatidylethanolamine isomers spin-labelled at six different positions along the acyl chain, and steroid spin labels, indicate both gel-fluid lamellar and lamellar-reverse hexagonal (HII) phase transitions, in agreement with complementary calorimetric studies. Analysis of spin label data shows that the transition to the HII phase is accompanied by an increase in conformational freedom of the acyl chain, more pronounced towards the methyl terminus, and representing an increase in the population of gauche isomers which can only be accommodated by a transition to the non-bilayer phase. Raising the bulk pH to, and above, pH 8.5 results in stabilisation of the bilayer phase and no transition to the HII phase is observed. The phosphatidylethanolamine spin labels also indicate a polarity profile which is characteristic of each phase. PMID- 6282592 TI - The assessment of an edge detection algorithm in determining left ventricular ejection fraction using radio-nuclide multiple gated acquisition and contrast ventriculography. AB - The application of an edge detection algorithm (EDA) in defining the boundary of the left ventricle (LV) in multiple gated (MG) cardiac studies has been assessed by comparison of the left ventricular ejection fraction (LVEF) derived from X-ray contrast ventriculography (CV) in a series of patients. The results demonstrate good correlation between methods irrespective of the projection and LV status when an EDA is used, such that the correlation in anterior and left anterior oblique (45 degrees) views is 0.91 and 0.86 respectively. PMID- 6282593 TI - Rapid radioimmunoassay for insulin and its application in localizing occult insulinomas by intraoperative pancreatic vein catheterization. AB - A rapid RIA for insulin and its application to localising occult insulinomas during surgical exploration is described. A standard curve and twelve serum samples can be assayed within 45 min. The assay is equivalent with respect to accuracy and precision to conventional RIAs for insulin. Pancreatic vein catheterisation at laparotomy and immediate insulin determination made it possible to localize and resect two insulinomas which were not demonstrated by arteriography, computed tomography, ultrasound, and not palpable at surgery. We suggest that the new technique will end the long controversy concerning subsequent management of occult insulinomas. PMID- 6282594 TI - Rapid miniaturized chromatography for Tc-99m IDA agents: comparison with gel chromatography. AB - Miniaturized chromatography systems for determining free pertechnetate and hydrolyzed reduced 99mTc levels in commercial 99mTc-labeled iminodiacetate (IDA) hepatobiliary radiopharmaceuticals were evaluated and the results compared with gel chromatography column scanning (GCS). Commercial IDA agents were evaluated including 2,6-dimethyl IDA, p-isopropyl IDA, p-butyl IDA, and 2,6-diisopropyl IDA. Of all the chromatography systems evaluated, only Gelman ITLC-SA with 20% NaCl and Gelman ITLC-SG with distilled water correlated with GCS in evaluating free pertechnetate and hydrolyzed reduced 99mTc levels for all IDA radiopharmaceuticals. The miniaturized chromatography procedure, as outlined, is rapid, taking less than 4 min, and can easily be incorporated into the daily quality control program in any nuclear medicine facility. PMID- 6282595 TI - Increase of plasma corticosterone induced by loperamide in rats. AB - Loperamide given intracerebroventricularly and intraperitoneally to rats provoked, like morphine, a plasma corticosterone increase 60 min after injection. Loperamide intracerebroventricularly was 3.73 times less active than morphine, while intraperitoneally it was 10.13 times more potent. This increase, associated with a significant elevation in the plasma ACTH concentration, was antagonized by naloxone (10 mg/kg i.p.) injected 30 min before loperamide. In hypophysectomized rats loperamide intraperitoneally did not affect the plasma corticosterone levels. We conclude that loperamide can stimulate corticosterone secretin from the adrenal gland via the opiate receptors and that this effect is mediated by a direct or indirect induction of ACTH release. PMID- 6282596 TI - Morphine-receptor dissociation constant and the stimulus-effect relation for inhibition of gastrointestinal transit in the rat. AB - The dissociation constant (KA) of morphine for its receptors was determined by the method of partial irreversible blockade of the receptor population using inhibition of gastrointestinal transit of a forced charcoal meal as the pharmacological endpoint. The anti-motility effect of morphine was antagonized when rats were pretreated with buprenorphine (0.3 mg/kg s.c.), a narcotic antagonist analgesic, 30 min before morphine and the extent of gastrointestinal transit was estimated a further 45 min later. With this schedule of drug administration, the agonist action of buprenorphine is minimal and its antagonist action predominates. The value of KA was (1.1 +/- 0.2) x 10(-5) mol/kg, a value close to that previously reported (2.9 x 10(-5) mol/kg) by us with these compounds in the rat tail flick test. The value of [A50], found here was 2.15 x 10(-6) mol/kg, approximately 1/5 of that of KA. Also, the stimulus-effect relation of the tissue, defined in Stephenson's theory, was plotted and found to be nonlinear. This result, when coupled with the inequality of KA and [A50], argues against the application of classical drug-receptor theory to this system. The apparent agreement between KA values for antinociception and inhibition of gastrointestinal transit is interesting, but does not necessarily prove equivalent receptors mediating the two different effects. PMID- 6282598 TI - MK 421 and prevention of genetic hypertension development in young spontaneously hypertensive rats. AB - MK 421, at the dose of 25 mg/kg, administered daily by gavage to spontaneously hypertensive rats (SHRs) from their 4th to 15th weeks of age almost completely inhibited development of genetic hypertension. Since heart rate and cardiac and systolic indexes were not affected by the drug, prevention of genetic hypertension development was solely related to an early, potent and long-lasting reduction of the progressive increase of the peripheral resistance which generally develops in SHRs during ageing. MK 421 reduced body growth but did not modify fluid intake, plasma NA+ and urine volume, thus water and salt retention did not develop. MK 421 enhanced vascular responsiveness to norepinephrine and angiotension II and reduced myocardial hypertrophy. Plasma renin concentration was increased and urinary antidiuretic hormone did not change. Finally, MK 421's preventive effects against genetic hypertension development persisted up to 10 weeks after discontinuation of treatment. PMID- 6282597 TI - Glucagon-induced densensitization: correlation between cyclic AMP levels and contractile force. AB - Desensitization of guinea pig left atria to the inotropic effects of glucagon was produced in vitro by treatment of the atria with 10(-6) M glucagon for 2 or 15 min. Desensitization of the contractile response was associated with a loss of the ability of glucagon to elevate cyclic AMP levels. A highly significant correlation was found between the increase in contractile force and the increase in cyclic AMP levels in control and glucagon desensitized left atria. PMID- 6282600 TI - In situ distribution of concanavalin A-binding sites in mesenchyme blastulae and early gastrulae of the sea urchin Lytechinus pictus. PMID- 6282599 TI - Lysosomal enzymes of Paramecium caudatum and Paramecium tetraurelia. PMID- 6282602 TI - Adenosine receptor activation in quiescent Swiss 3T3 cells. Enhancement of cAMP levels, DNA synthesis and cell division. PMID- 6282601 TI - Rough endoplasmic reticulum-mitochondrial complexes from rat liver. An extramitochondrial succinic dehydrogenase associated with this rough endoplasmic reticulum. PMID- 6282603 TI - Localization of vimentin and desmin in BHK21/C13 cells and in baby hamster kidney. PMID- 6282605 TI - DNA repair in cultured human fibroblasts does not decline with donor age. PMID- 6282606 TI - Fusion-induced differentiation of SV40-infected human keratinocytes. PMID- 6282604 TI - Effects of cyclic nucleotides on the shedding of tumor cell surface membranes. PMID- 6282607 TI - Triglyceride and cholesterol ester metabolism of rat 1 cells and Rous sarcoma virus-transformed rat 1 cells. PMID- 6282609 TI - Effects of vitamin A deficiency on [3H]retinoid binding to cellular retinoid binding proteins in rabbit cornea and conjunctiva. PMID- 6282610 TI - Cyclic nucleotide metabolism in inherited retinopathy in collies: a biochemical and histochemical study. PMID- 6282608 TI - pp60src-Dependent increase in adenosine-stimulated cAMP formation. PMID- 6282611 TI - Cation pump activity and membrane permeability in human senile cataractous lenses. PMID- 6282612 TI - Kinetics of calf lens phosphofructokinase activity at physiological substrate concentrations. PMID- 6282614 TI - Tissue factors influence fibroblast activity in neuromuscular diseases. PMID- 6282613 TI - Calcium-mediated myopathy at neuromuscular junctions of normal and dystrophic muscle. PMID- 6282615 TI - Activation of fibers via experimentally produced stump neuromas of skin nerves: ephaptic transmission or retrograde sprouting? PMID- 6282616 TI - Trypanosoma cruzi: ultrastructural cytochemistry of mitochondrial enzymes. PMID- 6282617 TI - Solubilization of prolactin receptor by a Zwitterionic detergent. PMID- 6282618 TI - Restriction enzyme studies on human highly repeated DNAs. PMID- 6282619 TI - Modulation of cis-platinum renal toxicity by the radioprotective agent WR 2721. PMID- 6282621 TI - Experimental coxsackievirus B4 valvulitis in squirrel monkeys. PMID- 6282620 TI - Cholestasis induced by scillaren administration, bicarbonate deprivation, or reduced hepatic blood flow. PMID- 6282622 TI - Effects of ischemia and pharmacological treatment on subcellular fractions from neonatal rat brain. AB - The effects of complete ischemia and of in vivo pharmacological treatment with trimetazidine were studied on some enzymatic activities related to energy transduction: lactate dehydrogenase for anaerobic glycolysis; citrate synthase and malate dehydrogenase for the Krebs' cycle; total NADH-cytochrome c reductase and cytochrome oxidase for the electron transport chain; glutamate dehydrogenase for amino acid metabolism and acetylcholine esterase for acetylcholine metabolism. These enzymatic activities were evaluated in brains of 10-day-old rats, at three different subcellular levels: homogenate in toto, purified mitochondrial fraction, crude, synaptosomal fraction. Complete normothermic post decapitative ischemia of 30 min duration increased the activity of cytochrome oxidase in the homogenate in toto and increased the activities of citrate synthase and malate dehydrogenase in the purified mitochondrial fraction, the activities of the enzymes evaluated in the crude synaptosomal fraction being unaffected. The i.p. treatment with trimetazidine (at the dose level of 50 mg . kg-1) was without any significant effect on the tested enzymatic activities. PMID- 6282624 TI - Synergistic activation of calcium-activated neutral protease by Mn2+ and Ca2+. PMID- 6282623 TI - Isolation of three native forms of myeloperoxidase from human polymorphonuclear leukocytes. PMID- 6282625 TI - Human heat shock gene expression and the modulation of plasma membrane Na+, K+ ATPase activity. PMID- 6282626 TI - Complex formation of guanidinated bovine trypsin inhibitor (Kunitz) with trypsin, chymotrypsin and trypsinogen as studied by the spin-label technique. PMID- 6282627 TI - The effect of colchicine on the in vivo and in vitro secretion of aldosterone by rat adrenocortical cells. PMID- 6282628 TI - An approach to the identification of adenosine's inhibitory site on adenylate cyclase. PMID- 6282629 TI - Solubilization of epinephrine-specific alpha-adrenergic receptors from adrenocortical carcinoma. PMID- 6282630 TI - Lipolysis of hepatic triacylglycerol stores. PMID- 6282631 TI - N-ethylmaleimide induces K+ -H+ antiport activity in Escherichia coli K-12. PMID- 6282632 TI - Lactose transport in Escherichia coli cells. Evidence in favor of a permease catalyzed efflux of lactose without protons. PMID- 6282633 TI - Activation, reduction and proton-deuterium exchange reaction of the periplasmic hydrogenase from Desulfovibrio gigas in relation with the role of cytochrome c3. PMID- 6282635 TI - Parts of the sequence between the complete rRNA operons are repeated on either side of the extra 16 S rRNA gene in chloroplast DNA of Euglena gracilis strain Z. PMID- 6282636 TI - Phosphorylation and activation of poly (A)-endoribonuclease from calf thymus gland. PMID- 6282634 TI - Homology of the 5'-terminal sequence of 28 S rRNA of mouse with yeast and Xenopus. Implication for the secondary structure of the 5.8 S--28 S RNA complex. PMID- 6282637 TI - Temperature-induced conformational changes in spin-labelled myoglobins: myoglobin of Aplysia brasiliana. PMID- 6282638 TI - In vivo cooperation between hepatic catalase and superoxide dismutase demonstrated by diethyldithiocarbamate. PMID- 6282640 TI - The influence of DNA gyrase on the transcription of linear DNA in vitro. PMID- 6282639 TI - Membrane glycoprotein synthesis: cleavage of the signal sequence in the absence of glycosylation. PMID- 6282641 TI - Isolation and characterisation of cyclic AMP-dependent phosphorylation sites from rat liver ribosomal protein S6. PMID- 6282642 TI - 31p NMR saturation transfer measurements of the steady state rates of creatine kinase and ATP synthetase in the rat brain. PMID- 6282643 TI - The EPR properties of nickel in hydrogenase from Methanobacterium. PMID- 6282645 TI - Formation of a conformationally changed C1r, a subcomponent of the first component of human complement, as an intermediate of its autoactivation reaction. PMID- 6282644 TI - Purine metabolism in Trichomonas vaginalis. PMID- 6282646 TI - Human complement subcomponent C2: purification and proteolytic cleavage in fluid phase by C1s, C1r2-C1s2 and C1. PMID- 6282648 TI - The effect of Ca2+ -mobilising hormones on the Na+ --K+ pump in isolated rat liver hepatocytes. PMID- 6282647 TI - Inhibition of anterior pituitary cyclic AMP phosphodiesterase by colchicine and vinblastine. PMID- 6282649 TI - Conformational changes of spin-labeled membrane proteins in human erythrocytes. PMID- 6282650 TI - Hormonal control of renal functions in insects. AB - Insect Malpighian tubules secrete an isosmotic, KCl-rich primary urine containing low concentrations of most other blood solutes. Neuropeptide diuretic hormones (DH), possibly related to vasopressin, stimulate tubular fluid secretion by 2- to 200-fold in response to water loading, e.g., feeding. DH acts on tubules through cyclic AMP (cAMP) to stimulate salt transport without measurable change in osmotic permeability. Changes in composition of tubular secretion after stimulation and the possible control of DH release are discussed. Most of the water, ions, and metabolites in tubular secretion are normally reabsorbed by active mechanisms in the rectum, where the urine may finally become either hyposmotic or strongly hyperosmotic to the blood. A newly discovered neuropeptide, chloride transport-stimulating hormone, controls (via cAMP) reabsorption of the principal salt by stimulating K-dependent, electrogenic transport of Cl- across the apical cell border. Passive net absorption of K+ is thereby enhanced. Diuretic and antidiuretic factors may control osmotic permeability of the rectal wall and thereby influence the osmotic concentrations of the rectal absorbate and final urine. The increased recycling of a KCl-rich fluid through the Malpighian tubule-rectal system after feeding probably serves to clear the body of unwanted substances ingested with, and produced by, metabolism of the meal. PMID- 6282651 TI - Endocrine and neural control of amphibian renal functions. AB - Three aspects of amphibian renal functions were considered. 1) The neurohypophysial hormone, arginine vasotocin (AVT), is diuretic in lungfishes, but is antidiuretic in amphibians. AVT probably produces diuresis in fishes by increasing systemic blood pressure and, hence, glomerular filtration rate. Receptors for AVT in early amphibians may have become more numerous or sensitive in the preglomerular circulation than in the peripheral vasculature. AVT could then produce glomerular antidiuresis. Tubular receptors to AVT also developed to provide better control of urine volume during terrestrial adaptation. In our investigations only glomerular antidiuresis was produced by AVT in the mud puppy whereas bullfrogs responded to AVT with both glomerular and tubular antidiuresis. 2) Although exogenous AVT can produce antidiuresis in amphibians, alpha adrenergic neural mechanisms also appeared to play an important role in glomerular and tubular functions in the bullfrog. 3) Mesotocin, the amphibian neutral neurohypophysial hormone, produces glomerular diuresis in the amphibians studied. However, whether it has a physiological role in regulating amphibian renal function remains unclear. PMID- 6282652 TI - Vascular smooth muscle in hypertension. AB - The cause of the elevated arterial pressure in most forms of hypertension is an increase in total peripheral resistance. This brief review is directed toward an assessment of recent investigations contributing information about the factors responsible for this increased vascular resistance. Structural abnormalities in the vasculature that characterize the hypertensive process are 1) changes in the vascular media, 2) rarefication of the resistance vessels, and 3) lesions of the intimal vascular surface. These abnormalities are mainly the result of an adaptive process and are secondary to the increase in wall stress and/or to pathological damage to cellular components in the vessel wall. Functional alterations in the vascular smooth muscle are described as changes in agonist smooth muscle interaction or plasma membrane permeability. These types of changes appear to play a primary, initiating role in the elevation of vascular resistance of hypertension. These alterations are not the result of an increase in wall stress and they often precede the development of high blood pressure. The functional changes are initiated by abnormal function of neurogenic, humoral, and/or myogenic changes that alter vascular smooth muscle activity. PMID- 6282653 TI - Phospho transfer enzymes: lessons from stereochemistry. PMID- 6282654 TI - Effect of alcohol on the heart and cardiac metabolism. AB - In this report the disturbances in biochemistry of the heart muscle exposed to alcohol are delineated. All elements of cellular substructures are affected. In plasma membranes, (Na+ + K+)-activated ATPase (EC 3.6.1.3) is inhibited. Mitochondrial damage consists in diminished respiratory function and calcium uptake and binding. High-energy phosphates remain intact. Alcohol also affects the malate-aspartate shuttle. Acetaldehyde, a metabolite of ethanol, has a direct effect on myocardial protein synthesis through microsomal inhibition; however, the development of cardiac hypertrophy is not affected. Malfunction of sarcoplasmic reticulum is evidenced by disturbances in calcium binding and uptake. Effects of ethanol on the contractile machinery are deficiencies in the turnover rate of chemical into mechanical energy (diminished Vmax), and in the number of cross-bridges formed (P0). It increases stiffness of series elastic elements. There is diminished fatty acid oxidation with increased esterification. The involvement of CoA synthetase (EC 6.2.1.1), palmityl-carnitine transferase (EC 2.3.1.7), and pyruvate dehydrogenase complex in disturbed fatty acid oxidation is not certain. The role of catalase in myocardial ethanol oxidation was examined. Ethanol activates myocardial catalase-H2O2 complex (EC 1.11.1.6). The biochemical basis of fetal alcohol syndrome is low hepatic alcohol dehydrogenase (EC 1.1.1.1) activity during fetal life. PMID- 6282655 TI - Effects of ethanol on ion transport in muscle membranes. AB - Alterations in the function of cardiac cellular membranes may contribute to the pathogenesis of abnormal cardiac function after both acute and chronic administration of ethanol. Both the sarcolemmal sodium pump-derived Na+,K+-ATPase and the calcium pump of the sarcoplasmic reticulum are inhibited by ethanol, although inhibitory effects at concentrations that can be reached in humans are seen only under special conditions. Abnormalities in both the Na+,K+-ATPase and calcium pump have been observed after chronic ethanol ingestion, and there is evidence that chronic ethanol ingestion reduces the sensitivity of these activities to ethanol added in vitro. The acute effects of ethanol on these membrane activities appear to be related to an increase in membrane fluidity, whereas the altered functional state of membranes produced by chronic ethanol ingestion may reflect alterations in membrane lipids, possibly associated with a decrease in membrane fluidity. PMID- 6282656 TI - Alcohol and the heart: theoretical considerations. AB - It is now well established that consumption of ethyl alcohol, both acute and chronic, exerts deleterious effects on the heart. Evidence is presented that the initial event that precipitates both acute and chronic changes reflects the physical effects of alcohol on membrane phospholipids and perhaps proteins. The presence of alcohol increases membrane fluidity, a condition that leads to an adaptive change in the phospholipid composition of the membranes, with resultant greater rigidity of the membranes. The effects of alcohol on the lipid bilayer of the plasma membrane, when combined with other nonspecific insults, may lead to a drastic increase in calcium permeability; the resulting calcium influx may cause cell necrosis and initiate irreversibly cardiomyopathy. It is likely that changes in membrane fluidity also exert profound effects on enzyme and transport activities of membrane-bound proteins. In addition, alcohol may interact directly with the hydrophobic regions of proteins. Such interactions may play an important role not only in membrane-bound proteins, but also in alcohol-induced changes in contractile proteins of the heart. It is suggested that, in general, the effects of alcohol are similar to those of other anesthetic agents, and that the elucidation of the pathogenesis of alcoholic cardiomyopathy may require a deeper understanding of the physical interaction among alcohol, phospholipids, and proteins. PMID- 6282657 TI - Epidermal growth factor action as a model for lymphokine function. AB - The experimental findings described here represent a general approach toward understanding the mechanism of action of polypeptide hormones and mediators by using epidermal growth factor (EGF) as the model system. A photoaffinity labeling method was used for identifying the cell-surface EGF receptor and for following its dynamic and migratory fate after binding to EGF. A different approach involving receptor insertion revealed that the EGF receptor can be transferred in a biologically active orientation from donor hepatic membranes to recipient receptor-negative mutant cells by a novel mechanism requiring no added fusogenic agent. These studies of the EGF receptor led us to probe into the transduction mechanism that relates hormone-receptor binding to its biological function. EGF receptor interaction in responsive cells was shown to lead to the intracellular generation of macromolecular protein activators of nuclear DNA replication. These molecular intermediates lead to the generation of an ultimate committed prereplicative state that is an innate characteristic and a global property of the whole cell. These studies of the EGF receptor and EGF-induced internal events serve to illustrate the types of systems that might be explored in future research on lymphokines and other such biologically relevant mediators of mitogenesis. PMID- 6282658 TI - Dehydration and fluid balance: central effects of angiotensin. AB - Central effects of dehydration are stimulated by osmotic stimuli, the reduced input of volume receptors, and angiotensin II. The subfornical organ (SFO) and organum vasculosum laminae terminalis (OVLT) have become accepted as putative receptor sites for angiotensin II in the brain. The exact quantitative relationship between the hours of water deprivation and the amount of angiotensin generated peripherally and whether that amount is sufficient to induce thirst centrally have not been established, but there is no question that when animals are dehydrated their angiotensin levels rise and the animals are thirsty. Attempts to block centrally the contribution of angiotensin II to thirst have been variable and cholinergic inputs have to be blocked at the same time. Various stimuli for thirst interact in a parallel fashion, and when one stimulus is blocked the other stimuli are still effective. Plasma angiotensin II may induce natural thirst, but how it enters the brain still remains to be explained. Although the SFO and OVLT have no blood-brain barrier, the blood supply to these organs acts as a limited perfusion system whereby blood-borne proteins cannot diffuse far from the capillary bed. A second set of receptors is found on the ventricular surface of the OVLT, as shown by fluorescence labeled angiotensin II. The connection between the SFO and OVLT was cut by discrete knife cuts. Drinking to angiotensin II intraventricularly was not significantly altered but the pressor response was reduced by 50%. These results can be explained by a circuit for drinking passing down below the level of the knife cut and a separate pressor pathway passing dorsally through the area that was cut by the knife. Thirst and pressor neural circuits beginning with angiotensin receptors could explain some of the data accumulated with the AV3V syndrome that occurs when the OVLT and nucleus medianas are destroyed. PMID- 6282660 TI - [Adenovirus infections in children]. PMID- 6282659 TI - The mechanism of phenylalanine hydroxylase. AB - The site of oxygen binding during phenylalanine hydroxylase (PAH)-catalyzed turnover of phenylalanine to tyrosine has been tentatively identified as the 4a position of the tetrahydropterin cofactor, based on the spectral characteristics of an intermediate generated from both 6-methyltetrahydropterin and tetrahydrobiopterin during turnover. The rates of appearance of the intermediate and tyrosine are equal. Both rates exhibit the same dependence on enzyme concentration. PAH also requires 1.0 iron per 50,000-dalton subunit for maximal activity. A direct correlation between iron content and specific activity has been demonstrated. Apoenzyme can be reactivated by addition of Fe(II) aerobically or Fe(III) anaerobically and can be repurified to give apparently native protein. Evidence from electron paramagnetic resonance implicates the presence of high spin (5/2) Fe(III). As a working hypothesis we postulate that a key complex at the active site may be one containing iron in close proximity to a 4a peroxytetrahydropterin. PMID- 6282661 TI - [Suppressibility of parathyroid function in primary hyperparathyroidism as estimated by nephrogenous cyclic AMP (author's transl)]. AB - The effects of calcium injection (3 mg/Kg/10 min) or oral calcium administration (calcium lactate 7.7 g) on plasma iPTH and Nephrogenous cyclic AMP (NcAMP) were studied in 6 normal controls and 13 patients with primary hyperparathyroidism. In the control subjects, plasma iPTH determined by a predominantly carboxyl-terminal antiserum was less than 0.3 ng/ml before and after both calcium loads, whereas 41 approximately 98% (mean 67%) of NcAMP was rapidly and uniformly suppressed to a level lower than the normal value. In 2 patients with primary hyperparathyroidism, iPTH was clearly reduced from 8.0 to 4.6 ng/ml and 1.6 to 0.96 ng/ml, respectively, by the calcium load. However, in the other 7 patients with primary hyperparathyroidism who showed only a slight elevation of iPTH: less than 0.3 approximately 0.9 ng/ml, the reductions in iPTH were not detected after the calcium load: less than 0.3 approximately 0.7 ng/ml. In contrast, 30 approximately 54% (1.02 approximately 3.85 nmol/dl GF) of NcAMP, which was greater than the diurnal variation, was suppressed after calcium injection in 5 patients with primary hyperparathyroidism (2 of 4 patients with urological, and 3 of 5 patients with chemical hyperparathyroidism). But NcAMP was not suppressed in all 4 patients with skeletal hyperparathyroidism including one with proximal renal tubular dysfunction whose basal iPTH was elevated markedly but reduced clearly by the calcium load. In general, suppression of NcAMP was followed by a decrease of phosphate excretion. On the other hand, even in a patient with primary hyperparathyroidism whose NcAMP was not suppressed at all after the calcium injection, calcium infusion (15 mg/Kg/3h) resulted in some (23%) decrease in NcAMP. Oral calcium administration resulted in responses which were almost the same as those produced by calcium injection. These results suggest that NcAMP provides a useful index in the parathyroid suppression test in patients with primary hyperparathyroidism, especially those who display a rather mild elevation of iPTH. This is not the case, however, in a few patients who show a marked elevation of iPTH and/or proximal renal tubular dysfunction. PMID- 6282663 TI - [Chondroid syringoma or mixed tumor of the skin (author's transl)]. PMID- 6282662 TI - [The role of hypothalamic amines in controlling prolactin surge in the pseudopregnant rat (author's transl)]. PMID- 6282664 TI - Klippel-Trenaunay-Weber syndrome. PMID- 6282665 TI - Inhibitory effect of prostaglandin A2 on Na+-K+-ATPase activity in synaptic plasma membrane of rat brain in vitro. AB - 1. The Na+-K+-ATPase activity in the synaptic plasma membrane of rat brain was inhibited by PGA2 in vitro. 2. PGA2 concentration required for 50% inhibition was 8.91 x 10(-5) M. 3. Inhibition mode of PGA2 for both ATP and Mg2+ was uncompetitive type. 4. PGA2 was competitive and noncompetitive with Na+ and K+, respectively. 5. The Ki values for PGA2 inhibition for ATP. Na+, K+ and Mg2+ were 6.0 x 10(-5), 2.7 x 10(-5), 10.8 x 10(-5) and 0.6 x 10(-5) M, respectively. PMID- 6282667 TI - Signal transduction in the cellular slime molds. PMID- 6282666 TI - Adenosine receptors in rat brain membranes: characterization of high affinity binding of [3H]-2-chloroadenosine. AB - 1. [3H]-2-chloroadenosine has been found to be a suitable ligand for the study of adenosine receptors in rat brain synaptic membranes. 2. Binding sites labelled by [3H]-2-chloroadenosine had a high affinity with a KD value of 23.5 nM. 3. Binding is heat sensitive, pH dependent and probably involves protein molecules. 4. The IC50 values for 2-chloroadenosine, adenosine, L-N6-phenylisopropyladenosine and D N6-phenylisopropyladenosine, N6-cyclohexyladenosine and adenosine-5'-N-ethyl carboxamide inhibition of [3H]2-chloroadenosine binding are in good agreement with the values obtained in studies of the ability of these compounds to inhibit adenylate cyclase, suggesting that [3H]-2-chloroadenosine binding sites reported here are comparable to the adenosine A1 receptor site. 5. There are regional differences in [3H]-2-chloroadenosine binding to brain membranes. 6. This difference is probably due to the discrepancies in the number of binding sites, and is probably not caused by changing affinities of receptors to the ligand. PMID- 6282668 TI - Glucocorticoid receptor of X. laevis: possible effect of phosphorylation on hormone binding. AB - The cytoplasmic glucocorticoid receptor of X. laevis liver has a high affinity for [3H]dexamethasone (Kd, 0.3 x 10(-8) M), and its binding specificity for a variety of steroids is similar to that found for mammalian glucocorticoid receptors. The ability of this receptor to bind [3H]-dexamethasone is stable at 0 degrees C but is rapidly lost at 10 and 20 degrees C. Alkaline phosphatase increases, whereas molybdate and tungstate decrease, the rate at which the binding activity is lost. These results are consistent with the loss of binding activity being due to dephosphorylation of the receptor. Binding of [3H]dexamethasone to the receptor does not alter the rate at which the binding activity is lost but does increase the stabilizing effect of molybdate. 100 mM molybdate lowers the apparent affinity of the receptor for [3H]dexamethasone, suggesting that molybdate can interact with the X. laevis glucocorticoid receptor. Addition of UTP, but not ATP, GTP or CTP, reactivates the receptor binding activity, which indicates that the receptor may be phosphorylated by a UTP-dependent protein kinase. PMID- 6282669 TI - Degradation of human choriogonadotropin in preovulatory rat granulosa cells and in follicular fluid. AB - The catabolism of human choriogonadotropin (hCG) in rat preovulatory granulosa cells and follicular fluid was studied in order to elucidate the mechanisms responsible for the removal of receptor-hCG complexes. Rats pretreated with pregnant mare serum gonadotropin (PMSG) were injected i.v. with 125I-labelled hCG with or without unlabelled hCG a few hours before the surge of endogenous preovulatory gonadotropin. The radioactive components of granulosa cells and of follicular fluid were characterized by filtration through Sepharose-6B gel. Granulosa cells catabolized receptor-bound 125I-hCG to hormone sub-units; however, the rate of catabolism in vivo was relatively slow. Receptor-125I-hCG complex also appeared in the follicular fluid. Binding of 125I-labelled hCG to receptor molecules of follicular fluid in vivo was time-dependent and it was inhibited by smaller amounts of unlabelled hCG than the binding, in vivo, of 125I labelled hCG to the receptors of granulosa cells. Both follicular fluid and granulosa cell homogenate, in vitro, catabolized 125I-labelled hCG to hormone sub units. PMID- 6282671 TI - Stimulation by thyrotropin, cholera toxin and dibutyryl cyclic AMP of the multiplication of differentiated thyroid cells in vitro. AB - Primary cultures of dog thyroid cells have been established. The cells originated from follicles and displayed differentiation characteristics of such cells: iodide trapping and organification, responsiveness of iodide organification and cyclic AMP accumulation to thyrotropin (TSH), induction of a two-dimensional follicular structure by TSH. TSH also stimulated the multiplication of these cells. The effect of TSH was detected with concentrations as low as 100 muU/ml and was reproduced with purified TSH. It was reproduced by cholera toxin (10 ng/ml) and dibutyryl cyclic AMP (10(-5) M). The data show that TSH, which stimulates the function of thyroid tissue, in vivo and in vitro, activates the multiplication of differentiated dog-thyroid follicular cells in primary culture, which suggests that this trophic effect is, partly at least, mediated by cyclic AMP. PMID- 6282670 TI - Gonadal steroid modulation of LHRH-stimulated LH secretion by pituitary cell cultures. AB - Enzymatically dispersed rat pituitary cells were grown in primary culture, and LHRH-stimulated LH secretion was measured. Testosterone (T) decreased and 17 beta estradiol (E) increased pituitary responsiveness to LHRH. The effect of E on LH secretion was partly due to an increase in LH content. There was a latent period of 12 h for E and 18 h for T between the onset of steroid treatment and the manifestation of steroid action. Neither steroid was required to be continuously present in order to exert its effects. After steroid withdrawal, the effect of T persisted for 72 h and that of E for more than 96 h. The actions of both steroids were blocked by protein-synthesis inhibitors. These results are consistent with the hypothesis that steroid effects rely on a mechanism involving alterations in protein synthesis; the affected proteins may be involved in the process of LHRH action. PMID- 6282672 TI - GnRH-receptors of rat pituitary is a glycoprotein: differential effect of neuraminidase and lectins on agonists and antagonists binding. AB - The interaction of 125I-labeled gonadotropin releasing hormone (GnRH) agonist, [D Ser-(t-Bu)6,des-Gly10-ethylamide]-GnRH, and antagonist [D-pGlu1,D-Phe2,D-Trp3,6] GnRH with rat pituitary membranes was studied. Their binding was affected differently by pretreatment of membranes with neuraminidase and by wheat-germ agglutinin. Pretreatment of the membranes with neuraminidase abolished the specific binding of both antagonist and agonist, in a dose-response manner, with the former being less affected. Wheat-germ agglutinin affected antagonist binding slightly (22% inhibition at 40 microgram/ml) but inhibited agonist binding more markedly (50% at 40 microgram/ml). This inhibitory effect was specific since it was readily reversed by N-acetylglucosamine. Other lectins, such as concanavalin A and soybean agglutinin affected the binding of both agonist and antagonist to only a small degree. These results suggest that the GnRH-receptor of rat pituitary is a glycoprotein which contains sialic acid residue and that GnRH agonists and antagonists bind differently to the same receptor. PMID- 6282673 TI - Calculating specific binding for prolactin receptor assays. AB - Specific binding of [125I]iodo-prolactin and receptor is usually calculated by subtracting the amount of bound radioactivity of assays containing excess unlabeled hormone (competed) from that of noncompeted assays. A proportional method to calculate specific binding of prolactin-receptor or other hormone receptor interactions is introduced as a replacement for the currently used subtraction method. The proportional method calculates specific binding by multiplying the ratio of free radioactivities of noncompeted and competed assays by the radioactivity retained in the competed assay and subtracting this value from the radioactivity retained by the noncompeted assay. In prolactin-receptor assays greater specific binding is seen when data is calculated by this proportional method rather than the subtraction method. The amount of this increase relates directly to the levels of specific and nonspecific binding. Additionally, the binding characteristics of prolactin and receptor in both equilibrium and kinetic studies are significantly different when specific binding is calculated by the proportional method rather than the subtraction method. PMID- 6282674 TI - Evidence that the porcine thyrotropin (TSH) receptor contains an essential disulphide bridge. AB - The effects of reducing agents on membrane bound and detergent solubilised porcine TSH receptors have been investigated. Both 2-mercaptoethanol and dithiothreitol appeared to inhibit the TSH-binding activity by a direct effect on the TSH receptor itself and Scatchard analysis suggested that this was primarily due to an alteration in TSH-binding capacity. In addition, some binding activity could be recovered by reoxidation of reduced receptor preparations. These results suggest therefore that the porcine TSH receptor contains an essential disulphide bridge. PMID- 6282675 TI - [Liver and sexual freedom (author's transl)]. PMID- 6282676 TI - [Ranitidine effects on 24-hour intragastric pH in normal individuals. Interest of continuous monitoring for selection of an optimal therapeutic regimen (author's transl)]. PMID- 6282677 TI - Combined hepatic angiography and percutaneous aspiration biopsy in the evaluation of primary hepatic neoplasm. PMID- 6282678 TI - Unusual contrast accumulations seen during percutaneous cholangiography. PMID- 6282679 TI - Effects of substituted benzimidazole (H 149/94) on gastric acid secretion in humans. PMID- 6282680 TI - [Aging of bone tissue]. PMID- 6282681 TI - [Ultrastructure of experimental adenocarcinoma of the rat stomach following long term administration of MNNG]. AB - Gastric tumours were induced in rats by orally administration of N-methyl-N' nitro-N-nitrosoguanidine (MNNG). The received adenomas and adenocarcinomas were investigated electron microscopically. In this paper we give a short description of main ultrastructural specialities of tumour cells, which can be derived from mucoid surface cells and mucous cells of antropyloric glands. PMID- 6282682 TI - Epinephrine-induced changes in the cyclic nucleotide content of fish melanoma cells. PMID- 6282683 TI - Structure and variation of human ribosomal DNA: molecular analysis of cloned fragments. AB - Eco-RI-A fragments of the human ribosomal RNA gene family from two types of tissue and three individuals were cloned in lambda vectors and compared by restriction enzyme digestion and electron microscopy. The EcoRI fragment A contains (i) 0.2 kb of the 3' end of the 18S rDNA, (ii) 2.5 kb of internal transcribed spacer and the 5.8S rDNA, and (iii) 4.6 kb of the 28S rDNA gene. All of the six cloned rDNA fragments isolated are identical by these analyses. Moreover, all contain a HincII site that is absent in about 50% of the rDNA identified by genomic blotting. Polymorphism in the nontranscribed spacer rDNA was studied in genomic blots of BamHI-digested DNA, using the 3' end of the 28S rDNA as a probe. The boundaries between the 18S rDNA, internal transcribed spacer, 28s rDNA, and external nontranscribed spacer were determined by R-loop analysis, further defining the organization of the ribosomal RNA precursor. PMID- 6282684 TI - Cloning of bacteriophage T5 DNA fragments. III. Expression in Escherichia coli mini-cells. AB - Use has been made of the mini-cell system to study polypeptide synthesis from cloned EcoRI, HindIII and PstI fragments of T5 DNA. The correlation of certain gene products with known genes has been established, as well as the physical mapping of genes not yet identified genetically. In some cases, it has been possible to demonstrate the presence of T5 promoters on the cloned DNA fragments. The design of experiments to avoid certain artifacts inherent in the use of the mini-cell system is discussed. PMID- 6282685 TI - A restriction map of the ribosomal RNA genes and the short single-copy DNA sequence of the pearl millet chloroplast genome. AB - The chloroplast rDNA genes of pearl millet (Pennisetum americanum) have been cloned and physically mapped. The chloroplast genome of the pearl millet contains two identical rRNA genes located on DNA sequences that are inverted with respect to one another and separated by 12 kb of single-copy DNA. The rRNA genes were positioned on a restriction endonuclease map by using as hybridization probes specific cloned rDNA sequences from the chloroplast DNA of the alga Euglena gracilis. The 16S and 23S rRNA genes were shown to be approx. 2 kb from one another, and the 5S RNA gene is immediately adjacent to the 23S tRNA gene. PMID- 6282686 TI - Nucleotide sequence of the argF regulatory region of Escherichia coli K-12. AB - The deoxyribonucleotide sequence has been determined for the regulatory region of the arginine F gene (argF) of Escherichia coli K-12. The location of the argF coding region was deduced by comparison of the DNA sequence to the sequence predicted from the primary structure of the N-terminus of the argF gene product, the subunit of the "F" isoenzyme of ornithine transcarbamylase. Transcription of the argF gene was found to initiate at a position approx. 40 bp preceding the N terminal codon for OTCase. Comparison of the region surrounding the origin of transcription with a computer-generated "model promoter sequence" revealed structural similarities between the two sequences, in particular, the promoter associated stretches known as the "Pribnow box" and "minus 35 contact site". Another feature noted for the argF promoter region was its extreme abundance of A : T nucleotide pairs. In the region preceding the start site for argF translation, a sequence was observed to be complementary to the 3' end of the 16S RNA component of the E. coli ribosome. Both the length and the nucleotide sequence of the argF leader region indicate that the argF gene does not contain an attenuator proposed to exist in other operons concerned with amino acid biosynthesis. PMID- 6282687 TI - Sequence variation in the LEU2 region of the saccharomyces cerevisiae genome. AB - The LEU2 regions present on yeast plasmid vectors come from two sources, a series of strains derived from S288c and strain M127. The LEU2 region from the S288c series contains a Tyl-17 element with its associated delta sequences and a small repetitive RNA gene while the LEU2 region from M127 which is present on pJDB248, lacks the Tyl-17 element, but carries a delta sequence and a small RNA gene. The various LEU2 plasmids currently in use vary with respect to these sequences depending on which restriction fragment from the region is present on the recombinant molecule. In addition, strain M127 contains three LEU2 homologous sequences that are represented by different EcoRI fragments and which segregate independently at meiosis. Therefore, there are at least four forms of the centromere-distal EcoRI fragment of the LEU2 locus in the Saccharomyces cerevisiae gene pool; these are 7.1 kb, 1.9 kb, 1.48 kb and 1.15 kb long. PMID- 6282688 TI - Comparison of IS1, IS2 and IS3 copy number in Escherichia coli strains K-12, B and C. AB - The number of copies of IS2 and IS1 in the chromosomes of five Escherichia coli K 12 strains and E. coli B and E. coli C has been determined by hybridization. Among these strains, IS1 copy numbers range from 4 to 19 and IS2 copy numbers range from 0 to 12. IS2 is present once in the E. coli B chromosome, but it is absent from E. coli C. The copy numbers of IS3 in the same seven E. coli strains range from 4 to 6. PMID- 6282689 TI - Solution hybridization using cloned, 5'-32P-labeled, double-stranded DNA to distinguish among closely related nucleic acids. AB - A rapid, sensitive, and specific solution-hybridization method is described that utilizes cloned, double-stranded DNA. The DNA is treated with restriction enzyme(s), and fragments are 32P-labeled at their 5' termini with polynucleotide kinase. A single fragment is partially purified by gel electrophoresis, denatured, and annealed with unlabeled RNA or DNA. The reaction mix is treated with S1 nuclease, precipitated with TCA, and the [32P]DNA counted. Hybridization is recognized only when the 32P label is associated with duplexes that are TCA precipitable. The specificity of the method was analyzed by annealing experiments with cloned, human globin DNAs. Restriction fragments labeled in the 3' untranslated region of human beta globin clones formed S1-resistant, TCA precipitable duplexes with beta globin DNA, but not with delta or gamma globin DNA. Thus, a major advantage of the method is that it can distinguish among homologous nucleic acids whose uniformly labeled cDNAs cross hybridize under moderately stringent conditions. This assay is as sensitive as the [3H]cDNA hybridization method, and it circumvents the requirement for purified mRNA as a template for [3H]cDNA synthesis. It also avoids a gel electrophoresis step, it is rapid and quantitative, and many samples can be simultaneously analyzed. PMID- 6282690 TI - Construction and characterization of three yeast-Escherichia coli shuttle vectors designed for rapid subcloning of yeast genes on small DNA fragments. AB - We have constructed three new subcloning plasmid vectors, pRC1, pRC2, and pRC3, derived from pKC7, which allow the rapid, single-step subcloning of yeast genes. Subcloning with these vectors utilizes a partial digestion with Sau3A to generate a quasi-random set of DNA fragments from the original plasmid. All three vectors contain a kanamycin resistance gene. Therefore, if the original cloned yeast DNA fragment is present in a vector that does not specify kanamycin resistance, the subclone pool can be propagated in Escherichia coli in the presence of kanamycin to select against parent plasmids that escaped restriction by Sau3A. Selection by complementation in yeast yields a collection of plasmids with smaller yeast DNA inserts containing the gene of interest. In the vectors pRC2 and pRC3, constructed from pRC1, the unique BamHI site is located within an intact tetracycline resistance gene, thus making it possible to screen bacterial transformants for those containing recombinant plasmid molecules. Vectors pRC2 and pRC3 also contain the yeast 2 micrometers DNA replication origin, and thus are more stable than plasmids carrying only the TRP1-associated replicator (ars1). PMID- 6282691 TI - Cloning of the human cytomegalovirus genome as endonuclease XbaI fragments. AB - Restriction enzyme XbaI DNA fragments that represent 99% of the sequences from the long and short unique as well as the repeat sequences of the human cytomegalovirus (CMV) genome have been cloned into bacterial plasmid pACYC184. The viral DNA sequences associated with the recombinant plasmids were analyzed by restriction mapping and by hybridization to fragments of authentic viral DNA. The relationship of the cloned viral DNA fragments to the XbaI physical map of the viral genome is demonstrated. Even though large recombinant plasmids ranging from approx. 39 to 1.8 kb were isolated, most if not all of the viral DNA fragments were stable during propagation in Escherichia coli HB101. PMID- 6282692 TI - A set of synthetic oligodeoxyribonucleotide primers for DNA sequencing in the plasmid vector pBR322. AB - Seven oligonucleotide primers complementary to the plasmid vector pBR322 at positions adjacent to five of the unique restriction endonuclease cleavage sites (EcoRI, HindIII, BamHI, SalI and PstI) have been chemically synthesized. The polarity of the primers is such that any DNA inserted at one or a combination of two of the above restriction sites may be sequenced by the chain termination method using one of the synthetic DNA primers. One of the primers for sequencing inserts at the PstI site of pBR322 is also complementary to the M13 phage vector designated bla6. This set of universal primers is useful for rapid sequence determination of DNA cloned into pBR322 or M13bla6. PMID- 6282693 TI - An avian tumor virus promoter directs expression of plasmid genes in Escherichia coli. AB - A sequence in the long terminal repeat (LTR) of avian tumor virus (ATV) DNA was shown to contain a promoter acute in Escherichia coli. For this analysis the bacterial promoters for the tetracycline (Tc) and neomycin (Nm) resistance genes were deleted from different plasmids and replaced with various fragments derived from the ATV DNA. Expression of the drug-resistant phenotype in the recombinant plasmids at levels comparable to or greater than those found with parental bacterial promotes was shown to be dependent on the presence of an intact sequence ranging from nucleotide +19 to -23 (relative to the cap site) in the ATV DNA. Comparison of the consensus bacterial promoter with the nucleotide sequence in this region revealed strong similarities. PMID- 6282694 TI - Specific-purpose plasmid cloning vectors. I. Low copy number, temperature sensitive, mobilization-defective pSC101-derived containment vectors. AB - Two cloning vector plasmids, pHSG415 (7100 bp) and a lambda phage cos site containing derivative (cosmid) thereof, pHSG422 (8760 bp), were constructed from a low copy number plasmid (pSC101) replicon to permit the propagation of cloned DNA segments at low gene dosage levels. Two features of the vectors, namely temperature sensitivity of replication and inability to be mobilized by conjugative plasmids, cause them to exhibit a high level of "biological containment". The essential characteristics of pHSG415 and pHSG422 may be summarized as follows: (1) their genome copy number is low (4--6 copies/chromosome); (2) their replication ceases at high temperature and they are rapidly lost from host cells grown at temperatures of 37 degrees C and above; (3) the relaxation nick site of pSC101, which is thought to be synonymous with its origin of transfer replication, is absent from the vectors; as a consequence, they are not mobilized to a significant extent by co-existing conjugative plasmids that are able to mobilize wild-type pSC101; (4) they contain unique insertion sites for DNA fragments generated by the following restriction endonucleases: EcoRI, XhoI, XmaI, HindIII and PstI; pHSG415 additionally contains single BamHI, BstEII and HincII sites and may also be used to clone PvuI generated fragments; (5) the plasmids confer upon their host cells resistance to chloramphenicol, kanamycin and ampicillin, and every unique cloning site, except those of BamHI and BstEII, is located within one of these antibiotic-resistance genes. PMID- 6282695 TI - Specific-purpose plasmid cloning vectors. II. Broad host range, high copy number, RSF1010-derived vectors, and a host-vector system for gene cloning in Pseudomonas. AB - Host-vector systems have been developed for gene cloning in the metabolically versatile bacterial genus Pseudomonas. They comprise restriction-negative host strains of Pseudomonas aeruginosa and P. putida and new cloning vectors derived from the high-copy-number, broad-host-range plasmid RSF1010, which are stably maintained in a wide range of Gram-negative bacteria. These plasmids contain EcoRI, SstI, HindIII, XmaI, XhoI, SalI, BamHI, and ClaI insertion sites. All cloning sites, except for BamHI and ClaI, are located within antibiotic resistance genes' insertional inactivation of these genes during hybrid plasmid formation provides a readily scored phenotypic change for the rapid identification of bacterial clones carrying such hybrids. One of the new vector plasmids is a cosmid that may be used for the selective cloning of large DNA fragments by in vitro lambda packaging. An analogous series of vectors that are defective in their plasmid-mobilization function, and that exhibit a degree of biological containment comparable to that of current Escherichia coli vector plasmids, are also described. PMID- 6282696 TI - Functional expression of the Herpes simplex virus thymidine kinase gene in Escherichia coli K-12. AB - The recombinant plasmid pAGO contains the Herpes simplex virus type 1 (HSV-1) thymidine kinase (TK) gene and consists of a 2-kb PvuII fragment of HSV-1 DNA inserted into the PvuII site of pBR322. A deletion mutant of pAGO, designated pMH110, has been isolated which removes the normal HSV-1 TK gene promoter but places the promoter of the pBR322 tetracycline-resistance (tetr) gene only about 400 bp from the translational start codon of the HSV-1 TK polypeptide. In contrast to pAGO, which transforms mouse LM(TK-) cells to TK+ but is only weakly expressed in TK- bacteria, pMH110 not only efficiently transforms LM(TK-) cells to TK+ but also enables TK- Escherichia coli K-12 cells to form colonies on selective plates containing 5-fluorodeoxyuridine (FdUrd) plus thymidine (dThd) and to exhibit fully restored ability to incorporate [3H]dThd into DNA. The levels of TK activity expressed by bacteria harboring pMH110 were about as high as those expressed by bacteria harboring plasmid pTK3, which contains the wild type E. coli TK gene. The TK activity expressed in bacteria harboring pMH110 was partially purified and shown to be HSV-1-specific by serological and disc PAGE analyses and by experiments demonstrating that this enzyme phosphorylated [125I]deoxycytidine. PMID- 6282697 TI - Nucleotide sequences of small ribosomal RNA and adjacent transfer RNA genes in rat mitochondrial DNA. AB - The nucleotide sequences of genes for small 12 S ribosomal RNA and two transfer RNAs (tRNAPhe and tRNAVal) and their common upstream region in cloned rat mitochondrial DNA were determined. This DNA sequence of 1709 bp extends between the HindII and HindIII cleavage sites in the EcoRI-A and EcoRI-D fragments, respectively. There is no long reading frame for protein synthesis starting from the HindIII site and extending all the way to the HindII site, or vice versa. The heavy strand was found to be the template for these RNA transcripts. The 5' and 3' ends of the 12 S ribosomal RNA gene are contiguous with the tRNAPhe and tRNAVal genes, respectively. The structural features of the two tRNA genes are significantly different from the standard prokaryotic tRNA pattern. Since similar contiguous natures of these genes in human (Crews and Attardi, Cell 19 (1980) 775; Eperon et al., Nature 286 (1980) 460) and mouse (Van Etten et al., Cell 22 (1980) 157) mitochondrial DNA have been reported, comparative studies on the structures of these genes were performed by computer analysis and the characteristic features are discussed. PMID- 6282698 TI - SphI restriction map of bacteriophage lambda DNA. PMID- 6282700 TI - Electron microscopic mapping of RNA polymerase binding sites on SV40 DNA. PMID- 6282699 TI - Variation in frequency of transformation by plasmid YRp7 in Saccharomyces cerevisiae. AB - Hybrid plasmid YRp7 (Escherichia coli plasmid pBR322 carrying a 1.4-kb yeast fragment containing the TRP1 gene) transforms two related haploid yeast strains (trp1) to Trp+ at frequencies per microgram DNA varying by two orders of magnitude. The diploid cross of these two strains is transformed at a frequency less than that of the low-frequency parent, indicating that high-frequency transformation is inherited in a recessive fashion. Segregant strains of two tetrads of this cross showed intermediate levels of transformability, suggesting the polygenic inheritance of transformation frequency. Levels of variation are changed extensively when frequencies are expressed as transformants per regenerated spheroplast colony, although the inheritance of higher frequency remains clearly recessive. Storage of spheroplast-DNA preparations at 4 degrees C increases the yield of transformants with some strains. PMID- 6282701 TI - The cloned EcoRI fragments of Autographa californica nuclear polyhedrosis virus DNA. PMID- 6282702 TI - Distribution of IS5 in bacteria. AB - Four different strains of Escherichia coli and several other bacteria were examined by Southern analysis for the presence of the insertion element IS5 and IS5-like sequences. Variations in the copy number, degree of homology and restriction pattern of the IS5-like sequence were found among the different organisms. The number and distribution of IS5 sequences do not appear to correlate with the evolutionary relationship of the bacteria in which they occur. PMID- 6282703 TI - Nucleotide sequence and genome organisation of filamentous bacteriophages fl and fd. AB - The DNA sequence of the filamentous phage F1, consisting of 6407 nucleotides, has been determined. When compared with the DNA sequence of the related filamentous phage fd (Beck et al., 1978), the f1 sequence is one nucleotide shorter and differs in 180 positions from the fd DNA. Only ten of these base exchanges cause amino acid exchanges in the known gene products. Most of the exchanges in f1 are the same as in M13 (Van Wezenbeek et al., 1980), showing a near identity of these two phage (there are only 59 nucleotide differences). Regulatory units for replication, transcription, and translation are in their essential parts identical in all three phage. PMID- 6282704 TI - Plasmid reference center registry of transposon (Tn) allocations through July 1981. AB - The criteria for transposable genetic elements (Cohen, 1976) were stated at the Cold Spring Harbor meeting in 1976 (Campbell et al., 1977) and defined and clarified further (Campbell et al., 1979). At that meeting the Plasmid Reference Center agreed to serve as a clearing house for continuous transposon registry. Interim lists have been previously published (Lederberg, 1978, 1980, Microbial Genetics Bulletin 44 : 31; ibid 48 : 90). A complete file of Tn elements is listed below. PMID- 6282705 TI - New restriction endonucleases from Flavobacterium okeanokoites (FokI) and Micrococcus luteus (MluI). AB - Two new restriction endonucleases have been isolated from Flavobacterium okeanokoites IFO12536 and Micrococcus luteus IFO12992 and named FokI and MluI, respectively. Based on analysis of the sequences around the restriction sites, the recognition sequences and cleavage sites of these endonucleases were deduced as below: FokI: (formula: see text). MluI introduces double-strand cleavages at unique sequences that are completely two-fold rotationally symmetric like most type II restriction endonucleases. FokI belongs to a class of restriction endonucleases that recognize specific but asymmetric nucleotide sequences and introduce staggered cleavages at appointed positions away from the recognition sequences. PMID- 6282706 TI - Bacteriophage T5 growth in Escherichia coli containing PstI fragments of the colicin Ib plasmid. AB - PstI restriction fragments of the colicin Ib (ColIb) plasmid have been cloned into the Apr gene of the pBR322 vector. Colicin-producing clones (Col+) all contained two common PstI (L and U) fragments, 2000 and 800 bp long, respectively. All of these colicin producers were found to permit the normal growth of bacteriophage T5; the presence of the whole ColIb plasmid causes an abortive T5 or BF23 phage infection. None of the other clones selected for their inability to propagate T5 produced colicin. The clones (Abi+) that permitted only an abortive infection by T5 contained a single restriction fragment, O (1600 bp), which was not found in any of the colicin producers. Likewise, the specific fragments (L and U) found in the Col+ clones were not found in the Abi+ clones. These data are very hard to reconcile with the hypothesis of a colicin-induced cell deterioration after T5 phage infection. PMID- 6282707 TI - Mapping of the XhoI and SacI restriction sites on the bacteriophage T5 DNA. PMID- 6282709 TI - A new site-specific endonuclease Bbei from Bifidobacterium breve. PMID- 6282708 TI - Cloning of bacteriophage T5 DNA fragments. II. Isolation of recombinants carrying T5 PstI fragments. AB - The adjacent PstI-J, I and G fragments of the phage T5 DNA molecule (4.4, 4.6 and 7.2 kb, respectively) have been cloned in plasmid pBR322 and their locations verified by Southern blot analysis. The PstI I and G fragments overlap the previously cloned HindIII-P and G fragments and like those, contain no known genetic markers. In addition, one of the 12 newly isolated T5 mutants maps in this PstI-IG region. Thus, the size of the "empty" region between genes D15 and D17, which we have previously observed on the genetic map, extends to at least 11.8 kb. In contrast, the PstI-J fragment carried part of the D12 gene and the intact D14 and D15 genes. This clone is of particular interest since the D15 gene product is a nuclease and is responsible for the positive control of late gene transcription. The orientation of these genes relative to the T5 DNA molecule has been determined by a combination of restriction, deletion and complementation analyses. PMID- 6282710 TI - Organization of the ribosomal RNA gene cluster in Aspergillus nidulans. AB - DNA coding for ribosomal RNA in Aspergillus nidulans was found to consist of a unit 7.8 kb in size which is tandemly repeated in the genome and codes for 5.8S, 18S and 26S rRNA. The repeat unit has been cloned, and its restriction map and the location of the individual rRNA coding sequences within the unit have been established. PMID- 6282711 TI - The nucleotide sequence of cDNA coding for the structural proteins of foot-and mouth disease virus. AB - The complete nucleotide sequence of cDNA coding for the structural capsid polypeptides of foot-and-mouth disease virus (FMDV) (strain A(10)61) has been determined. Portions of the flanking sequence coding for the nonstructural proteins p20a and p52 are also provided. The three larger structural polypeptides VP1, VP2 and VP3 have unmodified Mrs of 23248, 24649 and 24213, respectively. The size of the smaller polypeptide, VP4, can only be estimated at 7360 because the 5'-limit of its coding region is not yet known with certainty. The sequence data for VP1 (the major immunising antigen) and the amino-terminal quarter of p52 are compared with the data of Kurz et al. (Nucl. Acids Res. 9 (1981) 1919-1931) for a different serotype (O1K). This shows that variation is much greater in the region coding for VP1 than in that coding for p52. This is reflected in the level of amino acid sequence variation predicted for the two proteins. Analysis of relative codon usage reveals a strong bias in favour of C and G over U and A in the third base position. The dinucleotide frequencies show a bias against A-U and U-A, and for A-C and C-A. PMID- 6282712 TI - Modification of Eco RI restriction sites by Caulobacter vibrioides. PMID- 6282713 TI - A modified pBR322 vector with improved properties for the cloning, recovery, and sequencing of blunt-ended DNA fragments. AB - The construction of a plasmid vector which facilitates the cloning and recovery of blunt-ended DNA fragments is described. This plasmid, called pHP34, differs from pBR322 by a 10-bp insertion which introduces a unique SmaI site immediately flanked by two EcoRI sites. Blunt-ended DNA fragments cloned in the SmaI site can be recovered by digestion with EcoRI. Small cloned fragments can be chemically sequenced using a strategy which does not require their purification. The use of a plasmid related to pHP34 for in vitro mutagenesis by the insertion of a DNA linker fragment conferring an antibiotic resistance is also discussed. PMID- 6282714 TI - DNA sequences flanking an E. coli insertion element IS2 in a cloned yeast TRP5 gene. AB - The insertion of an Escherichia coli IS2 element upstream from a cloned yeast TRP5 gene results in an increased level of active tryptophan synthase in trpAB E. coli host cells. This insertion occurs about 60 bp upstream from the first AUG of the TRP5 gene and is associated with a duplication of the sequence TTACA at the target site. The nucleotide sequence corresponding to the first 173 amino acids of the yeast TRP5 gene has also been determined. The N-terminal region of the yeast tryptophan synthase includes areas of strong homology with the alpha subunit of the corresponding E. coli enzyme. Sequences from the 5' untranslated region upstream from the TRP5 gene are compared to homologous areas of other yeast genes. PMID- 6282715 TI - Molecular cloning of the gene for dihydrofolate reductase from Lactobacillus casei. AB - The Lactobacillus casei gene for dihydrofolate reductase has been cloned in Escherichia coli using the multicopy vector pBR322. A restriction map of the cloned DNA has been prepared. The cloned DNA directs the synthesis of L. casei dihydrofolate reductase in E. coli and confers trimethoprim and methotrexate resistance. PMID- 6282716 TI - [Food value of vegetable and melon crops grown using different amounts of mineral fertilizers]. PMID- 6282717 TI - [Removal of various types of quartz and coal dust from the lungs of rats]. PMID- 6282718 TI - Differential effects of folic acid on water content, protein and microsomal 5' phosphodiesterase activity of the rat kidney. AB - The effects of folic acid administration on the weight, protein, water content and microsomal 5'-phosphodiesterase of the rat kidney were determined, to elucidate the mechanisms contributing to the renal enlargement produced by this agent. Folic acid administered ip in single doses of 100-250 mg/kg caused dose related increases in kidney weight, water and protein content within 24 hr. Time course studies indicated that 250 mg folic acid/kg given ip produced a progressive elevation in renal water content from 2 to 72 hr. Smaller increases in whole-kidney protein were recorded 8, 24 and 72 hr after folic acid treatment. However, a biphasic response of microsomal 5'-phosphodiesterase was produced, inhibition at 16 hr being followed by stimulation (to 140% of control) at 72 hr. In vitro studies indicated that folic acid inhibits 5'-phosphodiesterase competitively, and the early inhibition of 5'-phosphodiesterase in vivo appears to be due to a direct effect of folic acid on the enzyme. PMID- 6282719 TI - Presence of benzo[a]pyrene and other polycyclic aromatic hydrocarbons in suntan oils. AB - Column and thin-layer chromatography followed by spectrophotofluorometry were used to determine the carcinogen benzo[a]pyrene and other polycyclic aromatic hydrocarbons (PAH) in commercial samples of four suntan iols based on mineral and/or vegetable oils liable to contain traces of PAH. The analyses showed that all the samples contained benzo[a]pyrene together with three other mutagenic, co carcinogenic or non-carcinogenic PAH (perylene, fluoranthene and benzo]k]fluoranthene). One sample also contained the carcinogen anthanthrene. The total PAH content of the samples varied from 88 . 5 to 188 . 7 ng/g while benzo[a]pyrene levels were in the 1 . 5-4 . 7 ng/g range. The results suggest that users of suntan oils may be exposed to low levels of potentially hazardous PAH; however, in comparison with many other cosmetics that are presumably based on similar oils, suntan oils are only used intermittently and for relatively short periods. PMID- 6282721 TI - [Peripheral polyneuropathy in chlorprothixene therapy. A clinico neurophysiological documentation (author's transl)]. PMID- 6282720 TI - [Bronchial muscle tonus: physiological regulation and therapeutic modification]. PMID- 6282722 TI - Resistance to iron accumulation and presence of hepatitis B surface antigen in preneoplastic and neoplastic lesions in human hemochromatotic livers. AB - Two human cases of hepatocellular carcinoma combined with primary hemochromatosis and liver cirrhosis were studied with special reference to liver lesions displaying resistance to iron accumulation, and presence of hepatitis B surface antigen. Hepatocellular carcinomas, as well as small islands of hepatocytes within regenerative nodules, were free of the iron accumulation which otherwise occurred throughout the remainder of the hemochromatotic liver parenchyma. A positive reaction for hepatitis B surface antigen using the orcein staining method occurred randomly in iron-containing hepatocytes and in clusters of iron containing hepatocytes cirrhotic nodules, but completely iron-free cells of foci and carcinomas were negative for orcein. Therefore, these iron-free foci are suggested to be precursors to the carcinoma. PMID- 6282723 TI - The effect of hormones on lipoprotein lipase activity in skeletal muscles of the rat. PMID- 6282724 TI - Changes in maternal serum aldosterone, potassium and prolactin levels during beta receptor agonist treatment in third trimester pregnancies. AB - Beta-receptor agonist (terbutalin or isoxsuprin) administration to pregnant women either because of premature labor (therapy group; T; n = 8) or as prophylactic treatment (prophylaxis group; p; n = 8) resulted in a marked drop in serum potassium (p less than 0.001) during the first two hours of infusion in all patients. In five diabetic women in the P group the prolactin was significantly decreased (P less than 0.01) as was serum aldosterone levels (p less than 0.01). These changes were not observed in the T group. The two groups of pregnant women responded with a significant increase in free fatty acids (FFA) and glucose but the magnitude and the time coarse differed. Thus in T women, FFA reached a peak after 30-60 minutes into treatment and then returned to baseline levels. A closer analysis revealed that this pattern was only obtained in patients receiving terbutalin. Among women given prophylactic treatment with terbutalin, a continuous increase in FFA was noted over the initial two hours (p less than 0.001). The increase in serum glucose was continuous in the two patient groups (p less than 0.001). It is suggested that beta-receptor agonists in diabetic women induces a dopaminergic type of response since serum prolactin levels but not TSH concentrations were affected. The possibility that an increased PGE2 synthesis at the expense of PGF2 alpha might mediate this effect of beta-receptor agonists in discussed. It is also suggested that an increased prostaglandin synthesis might interfere with aldosterone secretion. The possibility that falling serum potassium levels may activate dopaminergic systems via PGE2 synthesis is also emphasized. PMID- 6282726 TI - Role of adrenal steroids on the cardiac c-AMP response to isoprenaline in the rat. AB - The effect of adrenalectomy on the basal cardiac c-AMP level and the elevated c AMP level induced by intravenous administration of isoprenaline (10 mcg/kg) were determined by radio-protein assay. The basal cardiac c-AMP level was not altered by adrenalectomy. But the cardiac c-AMP responses to isoprenaline was significantly less in adrenalectomized rats, maintained with 1% sodium chloride solution for one week; this effect, however, disappeared when animals were maintained for 2 or 4 weeks. This could not be restored by acute administration of dexamethasone (100 mcg/rat). The possible reasons for these effects of adrenalectomy are discussed. PMID- 6282725 TI - The hypophyseal-adrenal axis in the protein-calorie malnourished rat. AB - Male rats were fed an 8% low protein diet for 30 consecutive days beginning at 20 days of age. On day 50, the effect of the diet was evaluated on the hypophyseal adrenal axis and the observations compared with those noted in age-matched controls fed a standard laboratory diet. The malnourished animals had significantly larger adrenal gland weights, relative to body size, and higher levels of serum corticosterone. Additionally, the number of ACTH-secreting cells in the pituitary glands of these rats was 63.8% greater than that present in the controls. These results suggest that ACTH secretion and adrenocortical physiology are stimulated under conditions of experimentally induced protein-calorie malnutrition. PMID- 6282727 TI - Hypercalcitoninaemia in medullary carcinoma of the thyroid and other malignancies: value of calcitonin as tumour marker. AB - Elevated plasma calcitonin was detected by radioimmunoassay in 19 of 99 unselected cancer patients (19%). The frequency of hypercalcitoninaemia in patients with tumours of lung, uterus, gastrointestinal tract, and urinary bladder was about 40%, in carcinoma of the breast 18%, and in carcinoma of the skin 4%. In 8 patients with bone metastases, 4 were hypercalcitoninaemic. None of 19 patients with non-malignant disorders had elevated plasma calcitonin. The highest plasma calcitonin level in cancer patients was 8200 ng/l. Urinary cyclic AMP was elevated in patients with hypercalcitoninaemia as compared with patients with normal plasma calcitonin. All 6 patients with medullary carcinoma of the thyroid had very high plasma calcitonin levels: more than 50000 ng/l in five and 30000 ng/l in one patient. Of the family members of these patients, raised plasma calcitonin was detected after stimulation with oral alcohol in 6 of 12. It is concluded that plasma calcitonin levels in excess of 10000 ng/l suggest the presence of medullary carcinoma of the thyroid, while levels below 10000 ng/l have a poor diagnostic value but should alert the clinician to look for a possible malignancy. PMID- 6282728 TI - In vivo Response of X-linked hypophosphatemic mice to calcitonin. AB - Other investigations, using the Hyp mouse as an animal model for the human disease X-linked hypophosphatemia (XLH), have demonstrated renal hypersensitivity to calcitonin (CT) using in vitro single renal tubule adenylate cyclase microassays. Renal hypersensitivity to CT may explain the elevated fractional excretion of phosphate (FE-P) and urinary 3'-5'-cyclic AMP (UcAMP) present in Hyp mice. This current study was designed as the in vivo counterpart to the in vitro experiments. CT dose-dependent hypocalcemia when data were constructed in normal and Hyp mice 18 hours after thyroparathyroidectomy (TPTX). Exogenous CT administered to TPTX normal mice resulted in dose-dependent hypocalcemia when data were expressed as the percent change from pretreatment. However, CT did not elicit a hypocalcemic response in TPTX Hyp mice at any dose. Only TPTX normal mice responded to CT with significant hypophosphatemia. FE-P and UcAMP were not significantly changed by CT in either genotype. In a different experiment, a large pharmacologic dose of CT was given to TPTX mice. This dose resulted in a significant but similar elevation of FE-P 1 hour after injection in both Hyp and normal TPTX mice. While UcAMP also rose significantly in both genotypes, the percent increase compared to controls was greater in Hyp mice. In summary, results from these in vivo experiments indicate that Hyp mice are not hypersensitive to physiologic doses of CT, and in fact they seem to be resistant to the hypocalcemic effect of the hormone. The greater increase in UcAMP in TPTX Hyp mice after a pharmacologic dose of CT may be the basis for the earlier in vitro results reported by others. We conclude that renal hypersensitivity to CT does not play a role in the etiology of XLH in the Hyp mouse. PMID- 6282729 TI - Epididymal fat cyclic-GMP levels during hyperthermia in normal and diabetic rats. PMID- 6282731 TI - CDC sets probe of Kaposi's in homosexuals. PMID- 6282730 TI - Humoral modulation of hepatic nuclear triiodothyronine receptors in the cross circulated rat. PMID- 6282732 TI - Pancreatic isleitis with coxsackie virus B5 infection. AB - Coxsackie group B virus infection may be responsible for some cases of insulin dependent diabetes mellitus. However, Coxsackie B5 virus is rarely implicated in this respect. The authors observed striking pancreatic islet cell damage in an infant who died of a Coxsackie B5 virus infection. The inflammatory response consisted of mononuclear cells, and the acinar tissue was completely uninvolved. PMID- 6282734 TI - Herpetic tracheobronchitis: immunohistologic demonstration of herpes simplex virus antigen. AB - A patient with no evidence of underlying malignancy or immunosuppression was found to have herpetic tracheobronchitis by bronchial brushing cytologic examination and bronchial biopsy. Herpes simplex type I antigens were localized in infected cells by means of the unlabeled antibody enzyme technique. Occasional cases of herpes simplex viral tracheobronchitis or pneumonitis have been reported in patients with severe burns or with malignancy. This is an unusual finding in otherwise health individuals. PMID- 6282733 TI - Obliterative cholangitis due to cytomegalovirus: a possible precursor of paucity of intrahepatic bile ducts. AB - A fetus at 24 weeks' gestation was found to have ascites and abdominal calcifications by ultrasonography. Pathologic examination after prostaglandin termination of pregnancy two weeks later revealed disseminated cytomegalovirus infection with inclusions in the lung, pancreas, kidney, and liver. A mononuclear inflammatory reaction was present in the portal areas of the liver and in the kidney, and gliosis was diffuse in the brain. The liver lesions were of greatest interest because most portal areas had no bile ducts. In other portal areas there was a combination of inclusions in epithelial cells of the few remaining interlobular ducts and intense duct-oriented inflammation. Thus, in fetuses surviving intrauterine cytomegalovirus infection, inflammatory destruction of intrahepatic bile ducts could manifest as chronic cholestasis and paucity of bile ducts later in infancy. PMID- 6282735 TI - Gingival granular cell tumor of the newborn (congenital "epulis"): ultrastructural observations relating to histogenesis. AB - The authors report the electron microscopic features of gingival granular cell tumors (GGCT) resected from two newborn girls. Although origin from odontogenic epithelium has been proposed, the ultrastructural findings strongly support histogenesis from stroma (mesenchymal) cells. Tissue assay for estrogen receptors was negative, but considering the marked predilection of GGCT for females, a hormonal factor (or factors) may still be important in the development of GGCT. Clinical and morphologic features distinguishing GGCT from granular cell "myoblastomas" are briefly emphasized. PMID- 6282736 TI - Real-time classification of multiunit neural signals using reduced feature sets. PMID- 6282737 TI - Probable macrophage origin of the lipopolysaccharide-induced cytostatic effect on intra-erythrocytic malarial parasites (Plasmodium vinckei). PMID- 6282738 TI - In vitro generation of splenic suppressor cells by trypsin. AB - Rat spleen cells were mitogenically stimulated with concanavalin A (Con A) or sodium periodate and cultured for 14-20 hr with trypsin. When these trypsin activated cells were co-cultured with fresh mitogenically stimulated cells, [3H] thymidine incorporation into the fresh cells was suppressed. Artifactual and trivial effects of trypsin (e.g. increased release of cold thymidine, decreased cell viability, and a change in thymidine transport) could not account for the suppressor effect. Suppressor activity was not affected by removing B cells or macrophages before trypsin activation nor by treating the trypsinized cells with mitomycin C. Suppressor activity could only be generated when the spleen cells were stimulated with Con A or periodate during culture with trypsin, and supernates from the trypsin-activated cells did not have suppressor activity. The physiological significance of the results and possible mechanisms of action of the suppressor cells are discussed. PMID- 6282739 TI - Effects of concanavalin A and Fc fragments of IgG on human monocyte cAMP content: modulation of monocyte secretory function by cAMP. PMID- 6282740 TI - Gastric potential difference: a reliable index of mucosal integrity? AB - The clinical significance and the diagnostic value of measurement of gastric potential difference are critically reviewed. The lack of correlation between behaviour of potential difference and state of the mucosal barrier, especially under the influence of agents inducing changes in gastric acid secretion, are discussed. PMID- 6282741 TI - The same genetic locus directs differentiation-linked expression of endogenous retrovirus gp70 on thymocytes and spleen cells in the mouse. PMID- 6282742 TI - Studies on an outbreak of Japanese encephalitis in Kolar district, Karnataka state in 1977-78. PMID- 6282743 TI - Japanese encephalitis epidemic in Uttar Pradesh, India during 1978. PMID- 6282744 TI - Mechanism of diethyl-dithiocarbamate induced elevation of parasitemia in Plasmodium berghei infection. PMID- 6282745 TI - Beta-adrenoceptors in intestine, atria and trachea of rabbit. PMID- 6282746 TI - Selective enrichment of NADPH oxidase activity in phagosomes from guinea pig polymorphonuclear leukocytes. AB - Recent studies have demonstrated that the activated NADPH oxidase, the enzyme responsible for the stimulation of O2 consumption with O2 formation during phagocytosis, is located in the plasma membrane of leukocytes. The present work deals with whether the activation induced by phagocytosis involves the enzyme of the entire membrane or only that of the portion of the membrane that interacts with the phagocytosable particle and forms the phagosome. The results presented show that the activity of the NADPH oxidase of phagosomal membrane, isolated by centrifugation of homogenates on discontinuous sucrose gradients, is increased 12.6-fold with respect that of homogenate. In contrast, the activities of 5' nucleotidase and of acid p-nitrophenyl phosphatase, enzyme markers of the plasma membrane not activated during phagocytosis and uniformly distributed on the entire membrane, are increased only about three-fold with respect to that of homogenate. These results indicate that during phagocytosis and activation of NADPH oxidase is a segmentary response that involves only the enzyme that forms the phagocytic vacuole. This fact is relevant for the function of toxic intermediates of oxygen reduction that are discharged in direct contact with the engulfed agent. PMID- 6282748 TI - Only the chemotactic subpopulation of human blood monocytes expresses receptors for the chemotactic peptide N-formylmethionyl-leucyl-phenylalanine. AB - Human peripheral blood monocytes comprise a subpopulation of 20 to 40% that is capable of responding to chemoattractants and a remaining subpopulation that cannot respond. We were able to obtain 99%-pure attractant-responsive monocytes by using a newly constructed separation chamber. The binding of the radioactive chemotactic peptide N-formylmethionyl-leucyl-[3H]phenylalanine to migrating and nonmigrating populations was then studied. The binding was saturable at room temperature in the presence of azide. Saturation occurred at 5 x 10(-8) M, and 50% of the maximal binding was obtained at 10(-8) M, the concentration that induced optimal chemotaxis. The nonmigrating monocytes did not bind the peptide under the same conditions, which shows that at least one reason for a nonresponsiveness to chemotaxin is apparently a lack of receptors. By Scatchard analysis we calculated an equilibrium dissociation constant ranging from 23 to 37 nM; the number of binding sites per cell ranged from 64,000 to 77,000. The binding was very rapid. Fifty percent of the optimal binding occurred at 3.5 min, and equilibrium was reached after 20 to 30 min. Chemotactic deactivation of the monocytes reduced the number of available binding sites by 60%. PMID- 6282747 TI - Characterization of mononuclear phagocytes from the mouse, guinea pig, rat, and man. AB - The present paper describes cytochemical, membrane, functional, and mitotic characteristics of monoblasts, promonocytes, monocytes, and macrophages of the mouse, guinea pig, rat, and man. For all of these species the results show that after staining for nonspecific esterase, with alpha-naphthylbutyrate as substrate, and for lysozyme, mononuclear phagocytes can be distinguished from other cells, e.g., T and B lymphocytes. However, it must be kept in mind that immature and mature granulocytic cells are also lysozyme positive. The presence of Fc and C receptors is dependent on the maturity of the cells and the duration of incubation in vitro; with respect to the former, an in vivo population of immature mononuclear phagocytes may have a lower percentage of positive cells than is the case in a mature population, and with respect to the latter, the percentage of positive cells rises during incubation. Phagocytosis of opsonized bacteria and red cells is a reliable criterion for the distinction between mononuclear phagocytes and other cell types, e.g. lymphocytes and fibroblasts. In all of the species studied, the majority of both immature and mature mononuclear phagocytes ingested particles opsonized with IgG; the proportion of phagocytosis of red cells via C3 receptors is usually very small. Incorporation studies with [3H] thymidine have shown that immature mononuclear phagocytes (i.e., monoblasts and promonocytes) divided and that monocytes and macrophages do not. The small number of macrophages that incorporate [3H] thymidine are immature mononuclear phagocytes which have very recently arrived in the tissues from the bone marrow. Comparison of mononuclear phagocytes in different organs of various species has shown unequivocally that these cells belong to one cell line, called the mononuclear phagocyte system. PMID- 6282750 TI - Structural analysis of electrophoretic variation in the genome profiles of rotavirus field isolates. AB - Detailed structural studies were undertaken on five isolates of bovine rotavirus which showed variability in the migration patterns of their genome segments on electrophoresis in polyacrylamide gels. The individual genome segments of each isolate were characterized by partial digestion of terminally radiolabeled RNA with a base-specific nuclease. This analysis showed that whereas mobility variations were always associated with detectable changes in nucleotide sequence, sequence changes at least as great as those found in segments showing electrophoretic mobility variations were also detected in segments showing no mobility variation. Evidence for the occurrence of genome segment reassortment between viruses in the field was obtained from analyzing the species 11 RNAs from these five isolates. The overall conclusion from these results is that great care is required in the interpretation of simple genome profile analysis of different isolates for epidemiological purposes and that classification of these viruses solely on the basis of genome electropherotype could be misleading. PMID- 6282749 TI - Receptor in group C and G streptococci detects albumin structures present in mammalian species. AB - The presence of albumin structures with the capacity to bind to a surface receptor in group C and G streptococci was studied in serum samples from 45 mammalian species representing 15 different orders, using an inhibition assay. The ability of animal sera to inhibit the uptake of radiolabeled human serum albumin by the streptococci indicated the presence of such albumin structures. Positive reactions were found in species of most orders tested, with Marsupialia as a notable exception. All Carnivora sera tested were strongly positive. In some orders such as Artiodactyla both positive and negative species were identified. Serum samples from 62 bird species representing 15 orders and from 5 fish species were also tested in the inhibition assay. None of these serum samples was capable of inhibiting the uptake of human serum albumin by streptococci. Some differences were also noted in the results obtained with group C and G streptococci from human and bovine sources, respectively, indicating the presence of two types of receptors. The present studies suggest a phylogenetic origin of albumin structures with affinity for the streptococcal receptor to a period after the divergence of Marsupialia from the other mammalian orders. PMID- 6282751 TI - Pyocin-resistant lipopolysaccharide mutans of Neisseria gonorrhoeae: alterations in sensitivity to normal human serum and polymyxin B. AB - Pyocins from Pseudomonas aeruginosa were used to select several lipopolysaccharide (LPS) mutants of Neisseria gonorrhoeae strain FA19. Three classes of LPS mutans were found in the initial group selected for study. The LPS of one class lacked galactose. That of a second group lacked the typical heptose found in the parental LPS, was reduced in glucose, galactose, and N acetylglucosamine content, appeared to contain a new unidentified sugar component, and consisted of two species of LPS separable on sodium dodecyl sulfate-polyacrylamide gels. The LPS of a third strain lacked the heptose, glucose, galactose, and N-acetylglucosamine found in the oligosaccharide portion of parental FA19 LPS. The minimal inhibitory concentration for polymyxin B of the mutant strains was 3 to 4 times that of the parental strain. The strains lacking only galactose were as resistant as the parent to the bactericidal action of normal human serum, but cells of the other two classes were quickly killed by serum. Gonococcal LPS thus appears to be important in determining phenotypic properties of the cells. PMID- 6282752 TI - Effect of immune serum or polymyxin B on Escherichia coli-induced inflammation and vascular injury. AB - Bacterial invasion of the tissues often stimulates a vigorous inflammatory reaction, which may limit the spread of microorganisms but may also be accompanied by serious vascular injury and tissue damage. We previously studied the inflammatory reaction induced by the injection of killed Escherichia coli into rabbit skin, a model suitable for the quantitation of various parameters of inflammation. Here we report the effect of immune serum treatment of the E. coli on their capacity to induce inflammation and vascular injury. Injection of killed E. coli treated with immune serum elicited a reaction which had a smaller increase in vascular permeability (protein exudation), measured with (125)I labeled albumin, less increase in blood flow, measured with (86)RbCl, less leukocyte infiltration, measured with (51)Cr-labeled leukocytes, and a lesser degree of hemorrhage, measured with (59)Fe-labeled erythrocytes, than E. coli treated with nonimmune serum. Crossover experiments with four different E. coli serotypes and four different antisera indicated that antibody to specific O antigens or a related antigen, but not to K or H antigen, was important for modifying the inflammatory response. Treatment of four different E. coli serotypes with antiserum to "core" glycolipid, produced by immunization with the E. coli J5 mutant, inhibited the inflammatory response to all four E. coli serotypes. Finally, treatment of killed E. coli with polymyxin B also inhibited their inflammation-inducing potential. These results suggest that it may be possible to diminish the magnitude of local vascular and tissue injury associated with E. coli infections by the use of antisera or polymyxin B, which bind to endotoxin on the E. coli. PMID- 6282753 TI - Evidence for the involvement of proton motive force in the transport of glucose by a mutant of Streptococcus mutans strain DR0001 defective in glucose phosphoenolpyruvate phosphotransferase activity. AB - Streptococcus mutans DR0001 and a glucose-phosphotransferase (PTS)-defective mutant, DR0001/6, were grown anaerobically in a chemostat with a glucose limitation at dilution rates (D) of 0.04 to 0.6 h(-1) (mean generation time, 17 to 1.2 h). The mutant possessed only 15% of glucose-PTS activity of the wild type and gave cell yields (19%) less than those of the wild type. Glucose-PTS activity in strains DR0001 was maximum at D = 0.1 h(-1) and was adequate to account for transport in the chemostat at all dilution rates except D = 0.6 h(-1), at which it was 80% of the actual glucose uptake activity. The mutant DR0001/6, on the other hand, possessed only sufficient glucose-PTS activity to sustain growth at below D = 0.1 h(-1), indicating the presence of an alternate transport activity. This was confirmed in glycolytic rate experiments with washed cells, which demonstrated that the mutant showed rates 11- to 27-fold higher than that accountable via glucose-PTS activity alone. The wild-type organism contained both a high (K(s), 6.7 to 8.0 muM)- and a low (K(s), 57 to 125 muM)-affinity transport system, whereas the glucose-PTS-defective mutant contained only the low-affinity system (K(s), 62 to 133 muM). The glucose-PTS was shown to be the high-affinity system. Glucose uptake by the mutant was unaffected by 8 mM sodium arsenate, 10 mM azide, and 10 mM dinitrophenol but was completely inhibited by 0.05 mM sodium iodoacetate. Glycolysis in the organism was almost completely inhibited by 0.25 mM N',N' -dicyclohexylcarbodiimide (DCCD), indicating the involvement of an ATPase in glucose uptake. The ionophores carbonylcyanide-m-chlorophenylhydrazone and tetrachlorosali-cylanilide were inhibitory at concentrations of 10 muM, suggesting that a proton gradient was important in the transport process. Higher levels of DCCD and the ionophores were required to inhibit the wild-type organism to the same degree. A mechanism is proposed for the alternative transport system whereby proton motive force is created by the extrusion of protons by the DCCD sensitive ATPase and glucose is transported down a proton gradient in a symport with protons. PMID- 6282754 TI - Degradation of intestinal glycoproteins by pathogenic Shigella flexneri. AB - Intestinal mucin glycoproteins were examined for their ability to sustain growth of pathogenic shigella. Inoculation of germfree cecal mucin glycoproteins with Shigella flexneri 4b resulted at 48 h in a 940-fold increase in the enteropathogen concentration. Investigation in vitro of enzymatic degradation by the pathogen led to the identification of a blood group B-degrading glycosidase produced by the bacteria. In in vivo experiments, fecal supernatants of mice monocontaminated with S. flexneri 4b contained an alpha-galactosidase active against the p-nitrophenyl-glycoside. This fecal alpha-galactosidase peaked 5 days after shigella contamination, showing 2.8 +/- 1.4 mU of enzyme activity per mg of protein. Contaminated fecal supernatants similarly destroyed the blood group B reactivity of cecal mucin glycoproteins. These data suggested that S. flexneri 4b could proliferate within ileocolonic environment by enzymatically degrading mucin glycoprotein sugars. PMID- 6282755 TI - Generation of cytolytic T-cell cultures displaying measles virus specificity and human histocompatibility leukocyte antigen restriction. AB - In the present study, peripheral blood lymphocytes from eight randomly selected, healthy, measles virus-seropositive donors were used to initiate and expand T cell cultures during secondary immune response in vitro. Five of the donors yielded continuously growing T-cell cultures which showed reproducible strong lytic activities towards measles virus-infected autologous fibroblasts. Uninfected or herpes simplex virus-infected targets were weakly susceptible to these effectors. By contrast, T-cell cultures from three other seropositive donors expressed comparable lytic activities for measles virus- or herpes simplex virus-infected targets, but not for uninfected autologous targets. The five T cell cytolytic cultures which revealed measles virus specificity also displayed human histocompatibility leukocyte antigen (HLA)-A and HLA-B restriction, i.e, were lytic for targets sharing HLA-A or HLA-B or both with them. Additionally, it was found that a monoclonal anti-HLA antibody (W6/32) could effectively block the measles virus-specific and HLA-A- and HLA-B-related lytic activities of these cytotoxic T-lymphocytes. The specificity of this blocking effect was reflected by the inefficacy of a monoclonal anti-HLA-DR antibody to block the cytotoxic T lymphocyte-mediated lysis. PMID- 6282756 TI - Macrophage antiviral activity: extrinsic versus intrinsic activity. AB - Peritoneal exudate cells from strains of mice both resistant and susceptible to challenge with mouse hepatitis virus strain JHM were examined for extrinsic and intrinsic antiviral activity. Thioglycolate-elicited and resident peritoneal cells from uninfected mice were able to suppress viral growth in a permissive cell. The active cell in both populations is an adherent, radiation-resistant, Thy-1.2 antigen- and Ia antigen-negative cell. The suppression of virus replication was not related to nonspecific cellular cytotoxicity directed against the permissive host cell, and no interferon was detected. The expression of extrinsic antiviral activity was not related to the ability of the host to resist mouse hepatitis virus infection by virtue of either age or genetic background. The expression of intrinsic antiviral activity, on the other hand, correlated with the ability of the host to resist virus challenge, indicating a characteristic distinction between these two in vitro mechanisms of macrophage mediated antiviral activity with regard to host resistance to viral infection. Further, the ability of a macrophage to support viral replication itself was independent of the ability of the macrophage to suppress virus growth in another cell. PMID- 6282757 TI - Activity of rabbit monocytes, macrophages, and neutrophils in antibody-dependent cellular cytotoxicity of herpes simplex virus-infected corneal cells. AB - Rabbit phagocytes were examined for their ability to kill target cells infected with herpes simplex virus by antibody-dependent cellular cytotoxicity. Two sources of rabbit corneal cells were used as targets: Staaten Seruminstitut Rabbit Cornea (a continuous cell line) and stromal keratocytes from the middle layer of corneas excised from New Zealand white rabbits. Peritoneal exudate and alveolar macrophages were found to be the most active effector cells, followed by blood neutrophils and monocytes. Peritoneal exudate and alveolar macrophages killed target cells with dilutions of antibody as high as 1:10,000. Monocyte antibody-dependent cellular cytotoxicity activity was absent in over one-third of the rabbits tested and was only weakly active in positive rabbits. In vitro aging of monocytes did not enhance activity. Antibody reactive with peritoneal exudate macrophages, alveolar macrophages, and blood neutrophil effector cells appeared 7 days after intracorneal injection of infectious herpes simplex virus. Results of these studies show that in vitro assays with a complete rabbit system (effectors cells, antibody, and target cells) can be developed to monitor herpetic disease and suggest an active role for rabbit phagocytes in cytotoxicity of herpes simplex virus-infected cells. PMID- 6282758 TI - Virus-infected colostral cell cytokine stimulation of human leukocyte natural killer cytotoxicity. AB - Natural killer cytotoxicity is an important antiviral defense mechanism. Human peripheral blood mononuclear cells cultured with herpes simplex virus (HSV) infected cells produced a cytokine. This substance stimulated adult natural killer cytotoxicity from 53.0 +/- 10.5% to 79.8% (P less than 0.01) against HSV infected target cells. These data resulted in a calculated cytokine-dependent cellular cytotoxicity (CDCC) value of 65.8%. Cytokine production was not stimulated by uninfected cells and was independent of the presence or absence of antibodies to HSV in sera of donors and mononuclear cells. Cells from human colostrum also produced an HSV-stimulated cytokine which mediated CDCC by using both adult (19.8 +/- 3.9%) and neonatal (18.6 +/- 3.4%) mononuclear effectors cells. Colostral cell cytokine production was also independent of donor HSV serology. Not all colostral cultures produced the cytokine, and in general colostrum-stimulated CDCC was lower than peripheral blood leukocyte-stimulated CDCC. Colostral cell cytokine stimulation of neonatal natural killer cytotoxicity may account in part for the increased nonspecific resistance of breast-fed infants to viral infection. PMID- 6282759 TI - Monoclonal antibodies of four different specificities for neutralization of type 1 polioviruses. AB - Four independent hybridoma clones have been established that produce neutralizing antibodies specific to type 1 poliovirus. Each clone produced antibody which neutralized a distinct set of type 1 test strains: (i) all 15 strains tested; (ii) the inducer strain only; (iii) predominantly wild strains; or (iv) all vaccine-related and some wild strains. PMID- 6282760 TI - Specific inhibition of contact sensitivity in the guinea pig following tolylene diisocyanate inhalation. AB - Guinea pigs were exposed by head only inhalation for 5 h to concentrations of approximately 1 ppm of tolylene diisocyanate (TDI) and 14 days later pinnal application of TDI on 3 consecutive days was followed by a dermal application of TDI after a further 7 days. The degree of contact sensitivity, measured 24 h later, showed that prior inhalation inhibited the skin reaction to TDI but it did not change the skin reaction to chloro-dinitrobenzene (DNCB) in guinea pigs sensitised and challenged similarly with DNCB. The inhibition lasted up to 9 weeks. However, it did not occur in guinea pigs after intranasal instillation (instead of inhalation) and only occurred after oral dosage with TDI when the dose was relatively high. Treatment with cyclophosphamide before TDI inhalation reversed the inhibition of contact sensitivity, suggesting an involvement of suppressor cells. PMID- 6282761 TI - Preparation of technetium-99-DMS renal complex in solution and its chemical and biological characterization. AB - 99Tc-DMS solution was prepared by double isotope labelling, purified by molecular sieving through Sepharose 2B column, and checked by chromatographic and spectrophotometric methods, as well as by assessment of its biodistribution in rats. The preparations, obtained in a series of nine experiments, showed high renal uptake (30-40% of injected dose per organ) and could be used for further investigations of Tc-DMS biochemical behaviour in kidney tissue at subcellular level. Radioactive concentrations of purified 99Tc-DNS preparations were 37-74 kBq/ml and absorption maxima at 412-425 nm (yellow complex). By application of 99mTc, 99Tc-DMS preparation in rats, good renal scans have been obtained. PMID- 6282762 TI - Monoclonal antibodies to Epstein-Barr virus-induced, transformation-associated cell surface antigens: binding patterns and effect upon virus-specific T-cell cytotoxicity. AB - Spleen cells from mice immunized with Epstein-Barr virus-transformed lymphoblastoid cells (EB-LCL) were used to generate monoclonal antibodies to cell surface antigens associated with the EB virus-transformed state. Radioimmune and immunofluorescence binding assays identified two antibodies, MHM6 and AC2, which reacted consistently with all EB-LCL tested, with a subpopulation of cells in some but not all EB virus genome-positive Burkitt lymphoma lines, but with none of a range of EB virus genome-negative cell lines of lymphoma or leukaemia origin. While MHM6 appeared to bind an EB virus-related antigen, AC2 bound some other cell surface antigen which was also found on a small subpopulation of cells in lymphocyte cultures stimulated with phytohaemagglutinin or with pokeweed mitogen. MHM6 and AC2 recognized single polypeptides with apparent molecular weights of 45 kd and 80 kd respectively as shown by sodium dodecylsulphate polyacrylamide gel electrophoresis (SDS-PAGE) analysis of 125I-labeled cell surface polypeptides immunoprecipitated with these antibodies. These polypeptides were induced on experimentally-infected B cells within 24 h of the expression of the EB virus nuclear antigen, EBNA, at a time known to coincide with the appearance of the lymphocyte-detected membrane antigen, LYDMA. However, saturating concentration of MHM6 and AC2 were unable to protect EB-LCL target cells from lysis by LYDMA-specific cytotoxic T cells in a chromium-release assay. PMID- 6282763 TI - Final case reporting from the Ugandan prospective study of the relationship between EBV and Burkitt's lymphoma. AB - A prospective epidemiological study was carried out in the West Nile District of Uganda from 1972 to 1979 in order to investigate the aetiological role of the Epstein-Barr virus (EBV) in Burkitt's lymphoma (BL). By 1976, fourteen BL cases had been detected among the 42,000 children originally bled in the study area. Testing of sera from BL candidates and neighbourhood controls showed that children who develop BL later have EBV/VCA titres several dilutions higher than their age- and sex-matched neighbours. This appearance of a strong EBV activity long before BL development was taken as evidence of a causal role of EBV in BL. In order to add to the unique material of pre-bled BL cases, BL detection was continued up to March 1979 when field work became impossible in Uganda. Two additional pre-bled BL cases were found during this extension of the study. The serological and virological evaluation of these additional cases showed that the EBV/VCA titres, but not the EA and EBNA titres, were about two dilutions higher in the BL candidates than in the controls. Hybridization assays showed that both lymphomas contained EBV/DNA in the tumour cells. These additional results thus confirm the findings in the first 14 cases and strengthen the epidemiological evidence for a causal role of the EBV in endemic BL. PMID- 6282764 TI - Enzymes of collagen synthesis and type III procollagen amino-propeptide in serum from Nigerians with hepato-cellular carcinoma and other malignant diseases. AB - Serum immunoreactive prolyl hydroxylase protein (S-IRPH), galactosylhydroxylysyl glucosyltransferase activity (S-GGT) and the amino-terminal propeptide of type III procollagen [S-Pro(III)-N-P] were studied in 24 patients with primary hepatocellular carcinoma, 18 with secondary liver neoplasms and 35 with other malignant diseases but no evidence of liver involvement; this latter group included 13 patients with Burkitt's lymphoma, 11 with breast cancer and 11 with other neoplasms. Control values were determined for 60 apparently healthy Nigerians, S-IRPH and S-GGT were above the upper normal limit, defined as the mean + 2 SD of the controls, in all the patients with primary hepatocellular carcinoma and all but one with secondary liver neoplasms, whereas only one S-IRPH value and three S-GGT values exceeded this limit in the patients with other malignant diseases. The mean S-Pro(III)-N-P was even more elevated than S-IRPH and S-GGT in the primary and secondary liver neoplasm cases, but was also elevated in other malignant neoplasms; about one third of the patients with no evidence of liver involvement had a concentration exceeding the upper normal limit. A high correlation was found between the values for the three assays both in primary hepatocellular carcinoma and in the whole series of malignant diseases. The data suggest that primary and secondary malignant neoplasms of the liver have a high rate of collagen synthesis. The three assays may be of some value in the diagnosis of primary hepatocellular carcinoma and secondary liver involvement in other malignant diseases, and in monitoring the treatment provided. PMID- 6282765 TI - Sera from patients with undifferentiated nasopharyngeal carcinoma contain a factor which abrogates specific Epstein-Barr virus antigen-induced lymphocyte response. AB - A unique association of Epstein-Barr virus (EBV) with the undifferentiated nasopharyngeal carcinoma (NPC) is a well acknowledged phenomenon. We report here the detection of a factor present in the sera of NPC patients which inhibits the blastogenic response of lymphocytes from EBV seropositive individuals to EB virions or soluble antigens. This lymphocyte-stimulation inhibitor (LSI) was found to be associated with the IgA fraction of the serum immunoglobulins. No inhibitory activity was detected in the sera and their immunoglobulin fractions from healthy (both EBV-seropositive and seronegative) individuals and patients with other carcinomas of the head and neck region. Interestingly, the IgA-LSI was absent in the sera of NPC patients who were successfully treated and remained in remission, while it was readily detectable in the sera of NPC patients in relapse, LSI-positive IgA fractions did not inhibit mitogenic response of lymphocytes to phytohemagglutinin. Taken together, the data presented suggest that LSI is a specific inhibitor of the response of sensitized lymphocytes to EBV antigens and that it may indeed represent a marker of great clinical significance regarding undifferentiated nasopharyngeal carcinoma, particularly for its prognosis. PMID- 6282767 TI - Activation of an endogenous mouse type C virus by UV-irradiated murine cytomegalovirus. AB - Infection of Kirsten sarcoma virus-transformed non-producer BALB-3T3 mouse cells with UV-irradiated mouse cytomegalovirus (MCMV) resulted in activation of a xenotropic type C virus detected by infectious center formation in permissive rat or mink cells. The levels of type C virus activated by MCMV were related to the UV dose applied. Under optimal conditions the frequencies of activation varied from 3.0 to 8.0 x 10(-4). The capacity of MCMV to activate type C virus was abolished by heat inactivation and by neutralization with specific antiserum against MCMV. Virus induction decreased under conditions of cell exposure to hydroxyurea or actinomycin D, inhibitors of DNA and RNA synthesis, respectively, with actinomycin D having a greater inhibitory effect. This suggests that both DNA and RNA synthesis are required for UV-MCMV induction of murine xenotropic virus. PMID- 6282766 TI - Migration inhibition caused by EBV-specific 48K subcomponent of EBNA and the associated 53K cellular protein. AB - Leukocytes from EBV-seropositive but not seronegative healthy donors responded with significant migration inhibition to the 48K subcomponent of the Epstein-Barr virus determined nuclear antigen (EBNA), known to carry the virally determined antigenic specificity. A concentration of 10 micrograms/ml was still effective while 5 micrograms/ml had no detectable effect. EBNA-associated cellular 53K protein had no effect by itself, but it potentiated the effect of 48K, even if the latter was added at the subliminal concentration of 5 micrograms/ml. The related 53K protein, isolated from EBV-negative human lymphoma cells, was also effective, whereas the corresponding murine-tumor-associated 53K had no potentiating effect. Immunization of mice with an extract of DNA-binding proteins from EBV-carrying Raji cells, known to contain both 48K and 53K, induced a significant macrophage migration inhibition response, to both human 48K and 53K. Murine 53K was ineffective, however. Human but not murine 53K increased the migration inhibitory activity of subliminal concentrations of 48K in the murine macrophage system as well. These findings suggest that human but not murine 53K may reconstitute with 48K (EBNA) to form a highly immunogenic complex. PMID- 6282768 TI - Cyclic AMP-dependent protein kinase content of murine T and B lymphocytes. AB - Murine splenic lymphocytes have been separated by non-activating procedures into enriched T and B cell populations. Cyclic AMP-dependent protein kinase (cAPK) activity has been measured in those separated cell types by histone phosphorylation. As in other tissues, lymphocyte cAPK activity was found to be predominantly a soluble enzyme. By an analysis of cyclic AMP (cAMP) activation of enzyme activity it was shown that: (1) the basal activities were indistinguishable between these major lymphocyte subpopulations, (2) the Ka values for cAMP activation were identical for the two cell types (24 nM cAMP), and (3) the Vmax for enzyme activity was higher in T (5.47 pmole/min/10(6) cells) than in B (3.58 pmole/min/10(6) cells) cells. DEAE-cellulose chromatography experiments with mixed and enriched splenic lymphocyte populations extended and amplified this quantitative difference between T and B cells. PMID- 6282769 TI - In vivo enhancement of mitogen-induced lymphocyte DNA synthesis by sodium diethyl dithiocarbamate (DTC). AB - Sodium diethyl dithiocarbamate (DTC), a low molecular weight sulphur compound, has been previously shown to be endowed of immunomodulatory properties. To get a better understanding of the mechanisms whereby DTC regulates the immune response, mitogen-induced lymphoproliferation was studied in spleen from guinea pigs treated with a single i.v. injection of DTC. The results clearly show that DTC enhances, after a lag period, DNA synthesis in T and B lymphocytes. DTC which appears not to act at the level of immuno-competent cells seems to modulate the immune responses through indirect pathways. PMID- 6282770 TI - Time--domain analysis in gated cardiac blood pool studies. AB - A technique is presented for producing functional images derived from equilibrium gated blood pool studies as a means of diagnosing cardiac disease. These functional images are based on characteristics associated with the time variation of the count rate (the time domain) at each point of the image matrix rather than on the Fourier transform of the time-activity curve (the frequency domain) which has gained recent attention. As examples of this method, we present images which display the statistical variance of the time-activity curve at each pixel, corrected for the expected contribution due to random statistical fluctuation, and images which display the time at which each pixel reaches its minimum count value. Variance and time-to-minimum images are comparable to Fourier amplitude and phase images, respectively, and have been found to be useful in facilitating the diagnosis of wall motion abnormalities. A major advantage of time-domain analysis is the wide variety of features of potential clinical significance which may be investigated. PMID- 6282771 TI - Improvement of blood pool imaging using in vivo labeling of red blood cells. PMID- 6282772 TI - Serological studies on swine influenza in Egypt. AB - 480 pig serum samples collected from November 1979-October 1980 and 200 human sera collected in the same period, nearly have been used in serological investigations. 52.5% sera were positive against swine influenza (HSW1N1) 10% only of human sera were positive against (HSW1N1) virus. By using (H3N2) human virus, 10.4% of pig sera were positive, while 41% of human sera were positive against this virus. PMID- 6282774 TI - Effects of canavanine treatment on herpesvirus morphogenesis in cultured cells. AB - L-Canavanine, a naturally occurring analog of arginine, effectively inhibited the morphogenesis of herpes simplex virus (HSV) and human cytomegalovirus (HCMV) when added at the time of infection, but allowed the expression of several cytopathic changes. Exposure to canavanine at progressively later times ultimately led to qualitatively normal virion maturation. Under no conditions were morphologically aberrant viral particles observed. However, HSV-infected cells treated at 3 h postinfection or later did contain distinctive cytoplasmic inclusions resembling HCMV dense bodies. Recovery experiments showed that HCMV-infected cells exposed to canavanine for 1-5 days could support normal viral morphogenesis when washed free of this agent. PMID- 6282773 TI - Retroviruses in feral mice. PMID- 6282775 TI - Antibody pattern to human cytomegalovirus in patients with adenocarcinoma of the colon. AB - The role of human cytomegalovirus (CMV) was examined according to serological patterns in 37 patients with adenocarcinoma of the colon (ACC). The sera were examined for the presence of IgG antibodies by the immunoperoxidase antibody to membrane antigens (IPAMA) method and by the complement-fixation (CF) test. Antibody determinations were also performed by the IPAMA method for three other members of the herpesvirus group: Epstein-Barr virus (EBV), herpes simplex virus (HSV) and varicella-zoster virus (VZV). Comparison groups included normal subjects, ACC patients treated with chemotherapy, and patients operated on for benign diseases. No significant difference was found in the geometric mean titers (GMTs) for CMV and the other herpesviruses in the sera of nontreated ACC patients when compared with the control groups. However, a significantly elevated antibody titer to CMV was found in chemotherapy-treated ACC patients by both the IPAMA and CF methods. In this group, elevated titers were found by the IPAMA method for EBV and HSV, but not for VZV. The significance of serological studies in elucidating the role of CMV in ACC patients is discussed. PMID- 6282776 TI - Pregnancy following cytomegalovirus mononucleosis. AB - A case of cytomegalovirus heterophile-negative mononucleosis seen in a primary care setting is reported. The patient was a young woman with fever that had persisted for 24 days, and with relative bradycardia, hepatosplenomegaly, suboccipital lymphadenopathy and a rash. Diagnosis was based on clinical and hematological findings which included relative and absolute lymphocytosis with atypical lymphocytes, elevation of serum transaminases and the presence of IgM specific cytomegalovirus antibodies with a significant rise of IgG-type antibodies as tested by indirect immunofluorescence. The patient's last menstrual period occurred 16 days after defervescence and she gave birth to an apparently normal female term infant with an Apgar score of 10 who has not shed virus in her urine during the first six months of life. PMID- 6282777 TI - [Chondroid syringoma]. AB - Chondroid syringoma is a rare benign appendage tumor of the skin. Histologically, the tumor is characterized by the presence of sweat gland elements surrounded by a cartilage-like material. The lesions are most frequently located around the nose. Clinically, they appear as solitary, firm, circumscribed, slowly growing intracutaneous or subcutaneous nodules. In the majority of cases the clinical diagnosis is either basal cell epithelioma or cyst. This paper calls attention to the characteristic clinical features of chondroid syringoma as observed in five patients. PMID- 6282778 TI - [Isolation of a penicillinase-producing strain of Neisseria gonorrhoeae in Aix-la Chapelle]. AB - A penicillin-resistant strain of Neisseria gonorrhoeae was isolated from a surveyor living in Aixla-Chapelle, who had an acute gonorrheal urethritis after a trip to Asia (Singapore and Bangkok). The susceptibility pattern for 20 antibiotics was tested in sugar diffusion tests, for five antibiotics in agar dilution tests, and for 12 antibiotics in serial dilution tests with determinations of MIC and MBC, respectively. A modified method for the determination of the MIC is described. This method is characterized by an increased clarity of interpretation, reproducibility and avoidance of subjective mistakes in interpretation. The penicillinase production was demonstrated using the clover leaf technique. The isolated strain showed the highest MIC and MBC values for penicillin (MIC 48 micrograms/ml, MBC 64 micrograms/ml) and ampicillin (MIC 96 micrograms/ml, MBC 96 micrograms/ml) and the lowest values for cefotaxim (MIC less than 0.125 micrograms/ml, MBC 0,25 micrograms/ml). PMID- 6282779 TI - [Coincidence of multiple cylindroma and trichoepitheliomas (Brooke-Spiegler Syndrome)]. AB - The coincidence of cyclindromas and trichoepitheliomas has been subject of a rather broad discussion for its principal pathogenetic aspects. Clinical cases, however, have only rarely been observed. Therefore, a case-report is given. Although the heredity of the Brooke-Spiegler syndrome seems well established by now, there is no anamnestic data in our case. PMID- 6282781 TI - Radon daughter exposure to uranium miners. AB - Radon exposures to U.S. uranium miners under present conditions average about 1.3 WLM per year approximately or equal to 60 WLM per full working lifetime. This is intermediate between (a) the lowest exposures for which there have been excess lung cancers reported among U.S. miners (120-240 WLM) and (b) average environmental radon exposures (16 WLM), so models based on these two situations are used to estimate expected effects on present uranium miners. In Model A, the loss of life expectancy is 45 days, the SMR (standardized mortality ratio) for lung cancer is 1.10, and the SMR for all causes between ages 18 and 65 is 1.013. In Model B these are 10 days, 1.03 and 1.002 respectively. It is shown that the radon exposures to miners are similar to those to millions of Americans from environmental exposure, and that miner health risks are comparable to those of other radiation workers. Their lung cancer risk from radon is 7-50 times less than their job-related accident mortality risk, and represents 0.7-4% of their total risk in mining. Miners suffer from many diseases with SMR very much larger than that for radon-induced lung cancer, and there are many other occupations and industries with far higher SMR for lung cancer than that from radon exposure to miners. PMID- 6282780 TI - [Prostaglandins, leukotrienes, anti-inflammatory substances and their importance in inflammatory reactions of the skin]. AB - Prostaglandins appear to play an important role as mediators of inflammation since they fulfill the major criteria. They possess powerful proinflammatory properties; they are present in increased concentrations in a wide range of proinflammatory lesions, and anti-inflammatory drugs inhibit their formation. The role of leukotrienes and other hydroxy fatty acid products is far less clear although their participation in cellular events seems probable. Steroid and nonsteroid anti-inflammatory drugs owe their activity at least in part to inhibition of biosynthesis of prostaglandins, although these two drug classes appear to act at different points in the pathways. It becomes increasingly clear that the oxygenation of arachidonic acid leads to the formation of a multiplicity of pharmacologically active fatty acids of which only a few have so far been identified. PMID- 6282782 TI - An evaluation of working Level measurements using a generalized Kusnetz method. PMID- 6282783 TI - Radon daughter equilibrium and unattached fraction in mine atmospheres. AB - Measurements of the equilibrium between radon and its daughters in mine atmospheres are presented. The situation in mines seems to be too complex to be described by any simple theoretical model. There is however a good correlation between the equilibrium factor, F, and the individual daughter ratios. This suggests that the individual daughter concentrations may be assessed by Kusnetz measurements of the equilibrium factor, F. Measurements by the wire screen method indicate that the dust concentration is the main influencing factor upon the unattached fraction of the radon daughters. The unattached fraction of the potential alpha-energy was lower than 0.05 in most mining situations. In areas with very clean air, the unattached fraction may however be higher than 0.3. Measurements indicated a good correlation between the unattached fraction of the potential alpha-energy and the unattached fraction of the individual daughter products. This suggests that assessments of the unattached fraction of the individual daughters may be obtained by using the Kusnetz method for measurements on the screen and back up filter. PMID- 6282784 TI - Unexpected 226Ra build-up in wet-process phosphoric-acid plants. AB - During an investigation of the distribution of radium and uranium over the different process streams in phosphoric-acid production and in the nitrophosphate route, a higher than average radiation level was detected in the vicinity of some piping and vessels near the gypsum filtres. This higher radiation level is caused by radium, which, as a decay product in the 238U series, is present in very low concentrations (1-50 pCi 226R/g) in the phosphates used for fertilizer production. Specific research later on established that there were detectable radiation levels around vessels and piping in other phosphoric-acid works as well. It turned out that in all factories the filter part, and especially the washing-acid section showed the highest radiation-level. In the piping of the gypsum filter, through which the washing acid is discharged, a scaling of bariumsulphate and calciumsulphate with a higher radiumconcentration (+/- 0.1 Ci/g) is formed. The radium ions appeared to be incorporated in the bariumsulphate lattice, hence the solubility of the radium precipitate is very low. The danger of internal pollution, for example in cleaning operations by radio-active intake or inhallation of radon is very small in the cases described. The risk of external radiation can be prevented by periodically cleaning (every 1 3 yr) the equipment in which the radio-active scaling is formed. PMID- 6282785 TI - Radon and thoron daughters in housing. PMID- 6282786 TI - Approaches to the diagnosis and treatment of tumors of the parapharyngeal space. AB - Tumors of the parapharyngeal space appear as painless massess bulging into the tonsillar, nasopharyngeal, or retromandibular area. Most are benign and represent a wide range of tumor growth, from parotid extension to intrinsic growth of nerves, blood vessels, and salivary gland tissues within or around the parapharyngeal area. The anatomy of this area and diagnostic procedures are discussed. Sinus films, tomograms of the skull base and lateral pharyngeal area, CT scans with simultaneous parotid sialograms, and angiograms are part of the evaluation of these rarely seen tumors. The approach to therapy is discussed. PMID- 6282787 TI - Chromosomal patterns in a benign human neoplasm, the mixed salivary gland tumour. PMID- 6282788 TI - A model for spontaneous mutation in Drosophila caused by transposing elements. AB - Transposing genetic elements make up a significant proportion of the DNA of at least some eukaryotes. One of the potential side effects of transposition is a high rate of apparently spontaneous mutation. In this paper we consider some of the evolutionary strategies that might be involved in the suppression of transposition, and we outline a model for the control of spontaneous mutation in terms of the regulation of transposition. Specific predictions are based upon the well-characterized genetics of the mutator system associated with hybrid dysgenesis in Drosophila melanogaster, which has many parallels with the expected behaviour of transposing elements. PMID- 6282789 TI - Localization of adenylate cyclase and 5'-nucleotidase activities in human thyroid follicular cells. AB - The localization of adenylate cyclase and 5'-nucleotidase activities in the follicular cells of adenomatous goiter and normal thyroid was studied by light and electron microscopy. Simultaneous biochemical measurement for both activities was carried out to confirm the histochemical findings. Adenylyl-imidodiphosphate (AMP-PNP) was used as an effective substrate for adenylate cyclase. The specificity of the adenylate cyclase reaction was also examined by adding oxalacetic acid or PCMB as an adenylate cyclase inhibitor, and by adding sodium fluoride or TSH as an adenylate cyclase stimulator to the reaction mixture. In the case of tissue from adenomatous goiter, a large amount of the reaction product of the adenylate cyclase activity was found uniformly in the apical and lateral plasma membrane and not in the basal plasma membrane. In the cases of normal thyroid, a small amount of the reaction product of adenylate cyclase activity was demonstrated, and only in the lateral plasma membrane of the follicular cells. On the otherhand, the histochemical localization of 5' nucleotidase activity was the same in adenomatous goiter and normal thyroid. The reaction product of 5'-nucleotidase activity was found predominantly in the apical plasma membrane of the follicular cells. The biochemical findings indicated that the activity of adenylate cyclase per gram tissue was approximately 2 times higher in the case of adenomatous goiter than that in the case of normal thyroid, while the 5'-nucleotidase activity in adenomatous goiter was in slightly higher level than in normal thyroid. Thus the histochemically demonstrable amount of adenylate cyclase and 5'-nucleotidase reflected the activity levels measured biochemically. The lack of demonstrable adenylate cyclase activity in the basal plasma membrane suggests the possibility that this structure may not play any important role in TSH reception. PMID- 6282791 TI - Optimal surgical therapy in stage I and II ovarian malignancies. PMID- 6282790 TI - The role of thoracic and cranial irradiation for small cell carcinoma of the lung. AB - Since 1974, 120 previously untreated patients with small cell carcinoma of the lung seen in Therapeutic Radiology at The Medical College of Wisconsin have been entered into one of 4 successive studies. Study I used thoracic irradiation (TI) alone (4500-6000 rad in 3-6 weeks) with chemotherapy at progression. Study II randomized patients with limited disease to TI (3000 rad in 2 weeks) plus either cyclophosphamide, doxorubicin, vincristine (CAV) or total body irradiation (TBI); patients with extensive disease received TI + CAV. Study III employed prophylactic cranial irradiation (PCI) plus CAV and withheld TI unless there was incomplete response or recurrence. Of 93 evaluable patients from the first three studies, 55 had limited and 38 extensive disease. Study I (37 patients) showed a 62% complete response (CR) rate; 43% failed in the chest, 14% had brain metastases, and the median survival was only 22 weeks in spite of a preponderance of limited disease patients. Study II (27 patients) showed a CR of 59%; 30% had brain metastases and the median survival was 48 weeks. Study II patients (29) had a 69% rate; 72% failed in the chest, 4% with PCI developed brain metastases, and the median survival was 50 weeks. In March, 1979, Study IV was initiated; patients receive PCI (2500 rad in 2 weeks) plus high dose CAV, methotrexate and leucovorin. After 6 cycles, consolidation TI (3750 rad in 3 weeks) is given to patients with complete response. Preliminary results with 27 patients treated on this study show a 67% CR rate, a 41% chest failure rate (but only 11% for the patients who received thoracic irradiation) and no intracranial failures, but a 13% extracranial CNS failure rate. PCI, TI and spinal irradiation may be necessary to maximize the probability of long term disease free survival. PMID- 6282792 TI - Precise positioning of patients for radiation therapy. AB - We have developed a number of immobilization schemes which permit precise daily positioning of patients for radiation therapy. Pretreatment and post-treatment radiographs have been taken with the patient in the treatment position and analyzed to determine the amount of intratreatment movement. Studies of patients in the supine, seated and decubitus positions indicate mean movements of less than 1 mm with a standard deviation of less than 1mm. Patients immobilized in the seated position with a bite block and a mask have a mean movement of about 0.5 mm +/- 0.3 mm (s.d.), and patients immobilized in the supine position with their necks hyperextended for submental therapy evidence a mean movement of about 1.4 mm +/- 0.9 mm (s.d.). With the exception of those used for the decubitus position, the immobilization devices are simply fabricated out of thermoplastic casting materials readily available from orthopedic supply houses. A study of day to-day reproducibility of patient position using laser alignment and pretreatment radiographs for final verification of position indicates that the initial laser alignment can be used to position a patient within 2.2 mm +/- 1.4 mm (s.d.) of the intended position. These results indicate that rigid immobilization devices can improve the precision of radiotherapy, which would be advantageous with respect to both tumor and normal tissue coverage in certain situations. PMID- 6282793 TI - Eradication of bovine leukemia virus infection from a high-prevalence herd, using radioimmunoassay for identification of infected animals. AB - Radioimmunoassay (RIA), using the virion glycoprotein antigen, was applied in an attempt to eradicate bovine leukemia virus (BLV) infection from a herd in which virtually all the adult cattle are infected. Considering that most calves born to BLV-infected cows are negative for BLV at birth and remain negative for the first several months of life, the eradication program was based on the identification and isolation of the BLV-free calves born to infected cows. Twenty-five calves raised on colostrum and milk from their infected dams were classified as BLV-free on the basis of negative results in the RIA at 6 to 8 and 9 to 11 months of age. These animals were maintained in either complete (10 calves) or partial (15 calves) isolation from infected cattle and were examined at regular intervals for BLV and BLV antibodies. With the exception of 1 calf in the group raised in partial isolation, the animals have remained free of BLV up to the time of the last evaluation, when they were 32 to 35 months old. At these ages, more than 90% of the nonisolated cattle in the herd are BLV-positive. The data also show that this eradication trial would have failed if, in the initial procedure used to classify the calves as BLV-free, the agar gel immunodiffusion test instead of the RIA had been used. Inasmuch as the 25 calves in this study were fed colostrum and milk from their dams, the fact that only 1 of the calves became infected during the 26 to 29 months of observation provides further evidence that milk-borne transmission of BLV is infrequent and perhaps inconsequential. PMID- 6282794 TI - Bovine herpes mammillitis in two New York dairy herds. AB - Bovine herpes mammillitis was diagnosed in 2 New York dairy herds. Lesions ranged from vesiculation and ulceration of large (up to 10 cm wide) areas of udder and teat skin to single small (2-3 cm wide) plaques of edema. Some lesions resembled "umbilicated pocks" characteristic of cowpox virus infections. Recently freshened heifers were the most severely affected; older cows and heifers with less turgid udders had milder lesions. In 2 cows, incurable mastitis developed. In other cows, the lesions healed by centripetal growth of epidermis into the lesions. Diagnosis was made by isolation of bovid herpesvirus 2 from lesions in both herds and by serum-neutralization testing. Virus isolated from a cow in 1 herd was injected into 9 members of the same herd and may have been responsible for the absence of lesions on these animals; all other members of the herd were affected. PMID- 6282795 TI - Radioisotope imaging for the evaluation of thyroid neoplasia and hypothyroidism in a dog. AB - An 11-year-old dog was diagnosed as having concurrent unilateral follicular thyroid carcinoma and hypothyroidism. Radioisotope imaging with technetium 99m as sodium pertechnatate identified the extent of thyroid tissue involvement. A combination of surgical resection and hormonal supplementation resulted in a favorable clinical response. PMID- 6282797 TI - Nitrogen metabolism in sheep fed protein sources of various solubilities with low quality roughages. AB - Four rumen-cannulated, mature crossbred wethers were used in a 4 x 4 latin square experiment designed to examine the effects on protein metabolism of protein sources with various solubilities fed with low quality roughages. Four diets were used: (1) grass hay plus oat straw (control), (2) control plus urea, (3) control plus soybean oil meal (SBOM) and (4) control plus sunflower oil mean (SFOM). The diets were isocaloric, isofibrous and isonitrogenous (1.60% except for the control, which contained 1.09% N). The protein solubility means determined by three solvents (Burroughs' mineral mixture, McDougal's artificial saliva and sodium chloride) were 23.7, 50.5, 18.2 and 24.2% for the respective diets. The average protein solubilities of SBOM and SFOM were 20.5 and 35.4%, respectively. Apparent crude protein digestibilites were higher (P less than .05) for diets with higher protein solubility. The Urea diet (highest protein solubility) resulted in higher urinary N excretion (P less than .05) and lower N retention than the SBOM and SFOM diets. N retention was similar for the animals fed the SBOM and SFOM diets. Ruminal NH3-N and plasma urea N levels were higher (P less than .05) in animals fed the urea diet. The protein source altered VFA concentrations but not C2 to C3 ratios of VFA. The SBOM and SFOM produced more branched chain fatty acids. These data illustrate that protein sources of higher solubility result in higher crude protein digestion and higher N loss in the urine. N balance was similar with SFOM and SBOM. SFOM and SBOM can be used interchangeably on an equal protein basis for sheep fed high roughage diets. PMID- 6282798 TI - Influence of preshipment management on the adrenal response of beef calves to ACTH before and after transit. AB - The influence of preshipment management, transit (965 km) and postshipment management upon the ability of the adrenal gland to respond to exogenous adrenocorticotropic hormone (ACTH) was evaluated in 60 beef calves (47 bulls and 13 steers). Adrenal response to ACTH treatment was quantified before shipment (PRE), about 24 h (POST) and 3 wk after transit (3 WK). Calves were assembled for 4 or 10 d before shipment and fed either a hay or a high energy diet (HE). The area under the plotted curve for the plasma corticoid concentration response to ACTH was used as an adrenal response test (ART). The feeding of hay during the first 4 d of assembly resulted in a greater (P less than .05) PRE ART value than feeding of the HE diet, but after 10 d of assembly, the ART was similar for animals on the two diets. The ART at 24 h after transit was similar to that before shipment. The HE group had a greater (P less than .05) ART than the hay group 3 wk after arrival, however, values were dependent upon postshipment management. Placement of calves on winter wheat pasture soon after arrival resulted in greater ART (P less than .05) values after the 3 wk recovery period than for calves that grazed native range for 10 or 20 d before grazing winter wheat pasture. These data indicate that the ability of the adrenal gland to respond to ACTH is increased at preshipment if the assembly period is 4 d compared to 10 d and the adrenal response at 3 wk after arrival is greater if a HE diet is fed during assembly instead of hay. PMID- 6282800 TI - The effect of inhibitors of beta-lactamases on beta-lactamase extracts and on intact cells. PMID- 6282799 TI - Carbohydrate fermentation in the large intestine of lambs. AB - In one of two experiments, six 4-mo-old wethers fitted with ileal and cecal cannulas were fed three levels of concentrate (0, 40 or 80% corn) with forage in a replicated 3 X 3 Latin square design. Chromic oxide was fed as a marker. Quantities of starch recovered at the ileum and digested in the large intestine increased (P less than .05) with increased starch intake. Quantities of acid detergent fiber (ADF) recovered at the ileum increased with increased ADF intake; however; ADF digestion (grams) in the large intestine was highest (P less than .05) when the 80% corn diet was fed. Cecal total VFA concentrations, butyrate molar proportions and lactic acid concentrations were highest (P less than .05), and cecal acetate to propionate (C2:C3) ratios and pH values were lowest, in lambs fed 80% corn. In Exp. 2, 14 wethers (4-mo-old) were fed a forage (alfalfa hay) or a concentrate (80% corn) diet containing chromic oxide as a marker. Digesta grab samples were obtained by surgical procedure. The amounts of starch recovered at the abomasum and ileum and the amounts digested in the rumen, small intestine and large intestine were greater (P less than .05) when lambs were fed the concentrate. Large intestinal total VFA concentrations and butyrate molar proportions were higher (P less than .01) when the concentrate diet was fed; pH values were lower (P less than .01). Lambs fed the concentrate diet had lower (P less than .01) large intestinal and ruminal C2:C3 ratios of VFA. PMID- 6282801 TI - Clinical experience with cefotaxime in hospitalized patients. PMID- 6282796 TI - Cochlear transduction: an integrative model and review. AB - A model for cochlear transduction is presented that is based on considerations of the cell biology of its receptor cells, particularly the mechanisms of transmitter release at recepto-neural synapses. Two new interrelated hypotheses on the functional organization of the organ of Corti result from these considerations, one dealing with the possibility of electrotonic interaction between inner and outer hair cells and the other with a possible contributing source to acoustic emissions of cochlear origin that results from vesicular membrane turnover. PMID- 6282802 TI - Steam distillation and gas-liquid chromatographic determination of triallate and diallate in milk and plant tissue. AB - A procedure based on steam distillation is described for the determination of residues of the thiocarbamate herbicides diallate and triallate. The herbicides are steam-distilled directly from aqueous suspensions of milk and plant samples and trapped in hexane. After column cleanup on either activated Florisil or silica cartridges, samples are quantitated by gas-liquid chromatography. Recoveries of diallate and triallate from milk, lettuce, peas, corn, canarygrass seed and straw, and flax straw ranged from 77 to 96%. PMID- 6282803 TI - Replication terminus of the Bacillus subtilis chromosome. AB - Bidirectional replication of the Bacillus subtilis chromosome terminates at a point on the circular chromosome which is symmetrically opposite to the replication origin. Since replication rates are similar in both "halves" of the chromosome, termination presumably occurs at the meeting point of the two replication forks. To investigate whether the DNA sequence of this region of the chromosome contributes to the termination event, we have determined the latest replicating region of a chromosome in which this DNA sequence is no longer symmetrically opposite to the origin. The merodiploid strain GSY1127 has a very large nontandem duplication (approximately 25% of the total chromosome length) in the left-hand half of the chromosome, so that size and symmetry of this chromosome are grossly different from those of normal strains. We have examined the replication order of genetic markers in this strain by measuring subtilis terminal marker for replication remains a terminal marker in the merodiploid, i.e., replicates later than a marker situated symmetrically opposite to the replication origin. These results were supported by replication orders determined by pulse-density transfer experiments during synchronous replication. The data obtained indicate that there is a preferred site for the termination of replication in the B. subtilis chromosome. PMID- 6282804 TI - Regulatory properties of a fructose 1,6-bisphosphatase from the cyanobacterium Anacystis nidulans. AB - A fructose 1,6-bisphosphatase (EC 3.1.3.11) (FBPase) was purified over 100-fold from Anacystis nidulans. At variance with a previous report (R. H. Bishop, Arch. Biochem. Biophys. 196:295-300, 1979), the regulatory properties of the enzyme were found to be like those of chloroplast enzymes rather than intermediate between chloroplast (photosynthetic) and heterotrophic FBPases. The pH optimum of Anacystis FBPase was between 8.0 and 8.5 and shifted to lower values with increasing Mg2+ concentration. Under the experimental conditions used by Bishop, we found the saturation curve of the enzyme to be sigmoidal for Mg2+ ions and hyperbolic for fructose 1,6-bisphosphate. The half-maximal velocity of the Anacystis FBPase was reached at concentrations of 5 mM MgCl2 and 0.06 mM fructose 1,6-bisphosphate. AMP did not inhibit the enzyme. The activity of the FBPase was found to be under a delicate control of oxidizing and reducing conditions. Oxidants like O2, H2O2, oxidized glutathione, and dehydroascorbic acid decreased the enzyme activity, whereas reductants like dithiothreitol and reduced glutathione increased it. The oxido-reductive modulation of FBPase proved to be reversible. Reduced glutathione stimulated the enzyme activity at physiological concentrations (1 to 10 mM).l The reduced glutathione-induced activation was higher at pH 8.0 than at pH 7.0. PMID- 6282805 TI - Two complementation groups mediate tetracycline resistance determined by Tn10. AB - The structural and regulatory tetracycline resistance genes of transposon Tn10 are located on a 2,700-base pair HpaI fragment. We have used eight tetracycline sensitive mutations in the 2,700-base pair fragment, cloned into two compatible plasmids, to demonstrate that two complementation groups are required for tetracycline resistance. By genetic recombination with plasmids containing the regulatory or structural regions for resistance, we have determined that both complementation groups reside within the structural region. The complementation groups, designated tetA and tetB, are proximal and distal, respectively, to the promoter for the tetracycline resistance structural region. The tetB mutations are in the portion of the structural region that is known to encode the 36,000 molecular-weight, inner-membrane TET protein. The levels of tetracycline resistance expressed during complementation suggest a complex interaction between the products of the tetA and tetB loci. PMID- 6282806 TI - Genetic and molecular characterization of Tn21, a multiple resistance transposon from R100.1. AB - Tge transposon Tn21 has been transposed from R100.1 to plasmid pACYC184 and, from the resulting recombinants, to plasmid R388. The sites of insertion and the orientation of the element in several pACYC184::Tn21 recombinants have been examined. Restriction enzyme analysis of these recombinants has resulted in a detailed map of Tn21; this is compared with the published maps of the relevant part of R100.1. Heteroduplex analysis has shown short inverted repeat sequences at the ends of the element. With various in vitro-generated deletion mutants of Tn21, the internal gene necessary for transposition (tnpA) was localized within the terminal 4.3 kilobases of the right-hand end of the element. Genetic analysis of transposition of Tn21 suggests that the process proceeds via cointegrates. Since the end products of transposition are simple recombinants of the element and the recipient replicon, Tn21 must contain a gene that codes for a resolvase type of activity (tnpR gene). PMID- 6282807 TI - Isolation and characterization of an Fe,-S8 ferredoxin (ferredoxin II) from Clostridium thermoaceticum. AB - A second ferredoxin protein was isolated from the thermophilic anaerobic bacterium Clostridium thermoaceticum and termed ferredoxin II. This ferredoxin was found to contain 7.9 +/- 0.3 iron atoms and 7.4 +/- 0.4 acid-labile sulfur atoms per mol of protein. Extrusion studies of the iron-sulfur centers showed the presence of two [Fe4-S4] centers per mol of protein and accounted for all of the iron present. The absorption spectrum was characterized by maxima at 390 nm (epsilon 390 = 30,400 M-1cm-1) and 280 nm (epsilon 280 = 41.400 M-1 cm-1) and by a shoulder at 300 nm. The ration of the absorbance of the pure protein at 390 nm to the absorbance at 280 nm was 0.74. Electron paramagnetic resonance data showed a weak signal in the oxidized state, and the reduced ferredoxin exhibited a spectrum typical of [Fe4-S4] clusters. Double integration of the reduced spectra showed that two electrons were necessary for the complete reduction of ferredoxin II. Amino histidine, and 1 arginine, and a molecular weight of 6,748 for the native protein. The ferredoxin is stable under anaerobic conditions for 60 min at 70 degrees C. The average oxidation-reduction potential for the two [Fe4-S4] centers was measured as -365 mV. PMID- 6282809 TI - Several copies of the same insertion sequence are present in alpha-hemolytic plasmids belonging to four different incompatibility groups. AB - Alpha-hemolytic plasmids belonging to four different incompatibility groups contained several copies (two to six) of the same DNA sequence. This sequence was capable of illegitimate recombination and thus behaved as an insertion sequence element. Its size (1.9 kilobases) and restriction enzyme cleavage map suggested that it was different from previously described insertion sequence elements. PMID- 6282808 TI - Role of exonuclease III in the base excision repair of uracil-containing DNA. AB - Mutants of Escherichia coli K-12 deficient in both exonuclease III (the product of the xth gene) and deoxyuridine triphosphatase (the dut gene product) are inviable at high temperatures and undergo filamentation when grown at such temperatures. In dut mutants, the dUTP pool is known to be greatly enhanced, resulting in an increased substitution of uracil for thymine in DNA during replication. The subsequent removal of uracil from the DNA by uracil-DNA glycosylase produces apyrimidinic sites, at which exonuclease III is known to have an endonucleolytic activity. The lethality of dut xth mutants, therefore, indicates that exonuclease III is important for this base-excision pathway and suggests that unrepaired apyrimidinic sites are lethal. Two confirmatory findings were as follows. (i) dut xth mutants were viable if they also had a mutation in the uracil-DNA glycosylase (ung) gene; such mutants should not remove uracil from DNA and should not, therefore, generate apyrimidinic sites. (ii) In the majority of the temperature-resistant revertants isolated, viability had been restored by a mutation in the dCTP deaminase (dcd) gene; such mutations should decrease dUTP production and hence uracil misincorporation. The results indicate that, in dut mutants, exonuclease III is essential for the repair of uracil-containing DNA and of apyrimidinic sites. PMID- 6282810 TI - Tn292l, a transposon encoding fosfomycin resistance. AB - The determinant of resistance to fosfomycin of the Serratia marcescens plasmid pOU900 was transposed into the plasmid ColE1 and into the plasmid RP4 in the absence of the RecA function of the host. In each case, the acquisition of fosfomycin resistance was correlated with the presence of a discrete piece of DNA, uniform in size and in restriction pattern, This new, 7.5-megadalton transposable element encoding resistance to fosfomycin has been called Tn2921. A preliminary map of the transposon is presented. PMID- 6282811 TI - Determination of the transcription direction of the exuT gene in Escherichia coli K-12: divergent transcription of the exuT-uxaCA operons. AB - The exuT gene of Escherichia coli, coding for the hexuronate transport system, was fused to lac genes by the use of Mu d(Apr lac) insertions (M. J. Casadaban and S. Cohen, Proc. Natl. Acad. Sci. U.S.A. 76:4530-4533, 1979). The method of chromosome mobilization with F' lac::Mu episomes (J. B. Zeldis, A. I. Bukhari, and D. Zipser, Virology 55:289-294, 1974) made it possible to determine the transcription direction of the exuT gene from the orientation of the Mu d(Apr lac) insertion in the fusion strains. Our results for a exuT-lac fusion strain suggest that the direction of transcription of other single gene operon exuT is clockwise on the standard E. coli map and confirm that the direction of transcription of uxaC is counterclockwise. The two close operons exuT and uxaCA are thus transcribed in opposite directions. PMID- 6282812 TI - Bacillus subtilis DNA gyrase: purification of subunits and reconstitution of supercoiling activity. AB - DNA gyrase from Bacillus subtilis 168 was purified by affinity chromatography on novobiocin-Sepharose and shown to consist of two subunits, A and B, with molecular weights of 100,000 and 85,000, respectively. The B subunits, which contains novobiocin-sensitive. ATPase activity, could complement the gyrA protein of Escherichia coli. No complementation was detected between the A subunit and the E. coli gyrB protein. PMID- 6282814 TI - Effect of phosphoglycerate mutase deficiency on heterotrophic and autotrophic carbon metabolism of Alcaligenes eutrophus. AB - Mutants of Alcaligenes eutrophus were isolated on the basis of their inability to grow on succinate as the sole source of carbon and energy. The mutants also failed to grow on other gluconeogenic substrates, including pyruvate, acetate, and citrate. Simultaneously, they had lost their capability for autotrophic growth. The mutants grew, but slower than the wild type, on fructose or gluconate. Growth retardation on gluconate was more pronounced. The mutants lacked phosphoglycerate mutase activity, and spontaneous revertants of normal growth phenotype had regained the activity. The physiological characteristics of the mutants indicate the role of phosphoglycerate mutase in heterotrophic and autotrophic carbon metabolism of A. eutrophus. Although the enzyme is necessary for gluconeogenesis during heterotrophic growth on three- or four-carbon substrates, its glycolytic function is not essential for the catabolism of fructose or gluconate via the Entner-Doudoroff pathway. The enzyme is required during autotrophic growth as a catalyst in the biosynthetic route leading from glycerate 3-phosphate to pyruvate. It is suggested that the mutants accomplish the complete degradation of fructose and gluconate mutase lesion. The catabolically produced triose phosphates are converted to fructose 6-phosphate which is rechanneled into the Entner-Doudoroff pathway. This carbon recycling mechanism operates less effectively in mutant cells growing on gluconate. PMID- 6282813 TI - Marker rescue by a homologous recipient plasmid during transformation of gonococci by a hybrid Pcr plasmid. AB - A 42-kilobase hybrid Pcr plasmid (pFA14) was formed when the naturally occurring 7.2-kilobase Pcr plasmid pFA3 was introduced by transformation into a competent gonococcal recipient containing the 36-kilobase conjugative plasmid pFA2 (Sox et al., J. Bacteriol. 138:510-518). Analysis of the structure of pFA14 showed that it was a stable recombinant between pFA3 and pFA2. The transformation efficiency of pFA14 was increased 300- to 10,000-fold by the presence in isogenic recipients of the homologous plasmid pFA2. The presence of a homologous plasmid in the recipient also markedly increased the likelihood of recovery of intact donor-size Pcr plasmids in the transformants. The presence of pFA2 had no effect on the competence of piliated or nonpiliated gonococci for transformation by either linear chromosomal DNA or a nonhomologous Pcr plasmid. Increased transformation efficiency of the hybrid Pcr plasmid pFA14 may have been due to recombination between the nicked or linearized donor plasmid and the homologous recipient plasmid (marker rescue). PMID- 6282815 TI - Receptors for clonidine in brain: insights into therapeutic actions. AB - Clonidine acts at specific receptor sites in the brain. These sites, i.e. adrenergic receptors, can be localized in the brain at the light-microscopic level by utilizing autoradiographic techniques. Autoradiographic studies revealed a striking differential distribution throughout the brain. These results provide an anatomical basis for understanding the actions of clonidine in the CNS. For example, the finding of high densities of alpha-2-receptors in the nucleus tractus solitarius and in the dorsal motor nucleus of the vagus is very likely related to its antihypertensive action. Also, the striking codistribution of alpha-2 receptors with opiate receptors in some areas could provide an explanation for the observation that alpha-2 drugs block opiate withdrawal. PMID- 6282816 TI - Central noradrenergic neurons: a locus for the functional interplay between alpha 2 adrenoceptors and opiate receptors. AB - The locus coeruleus (LC) contains the largest clusters of noradrenergic neurons in the brain. Single-cell recordings in rats show that LC neurons can be inhibited by clonidine acting via alpha-2 adrenoceptors; morphine and opiate peptides are also inhibitory but act via separate opiate receptors. During states of opiate withdrawal LC neurons become hyperactive; under these conditions clonidine can normalize activity via alpha-2 adrenoceptors even when opiate receptors are blocked. These single-cell studies provide a possible mechanism for the clinical efficacy of clonidine in relieving the signs and symptoms of opiate withdrawal in human addicts. PMID- 6282817 TI - The primate locus coeruleus and effects of clonidine on opiate withdrawal. AB - Studies of nonhuman primates suggested that the noradrenergic locus coeruleus (LC) may be involved as part of the neural substrate for a brain alarm function which includes attentiveness, arousal, anxiety, fear, terror, and the physiological correlates of these states. The studies compared the effects of electrical activation of tiny electrodes in the locus coeruleus with the effects of other agents and conditions which increased or decreased LC activity. The results suggested that the activation of the LC system is prevented by endogenous morphine-like substances and by opiates and that the opiate withdrawal syndrome is due to activation of this LC-noradrenergic system. Clonidine, which in low doses suppressed noradrenergic and LC activity, was therefore postulated to suppress opiate withdrawal signs and symptoms. The confirmation of this hypothesis in rats, monkeys and human subjects has added to the understanding of the mechanisms of opiate action and withdrawal. PMID- 6282819 TI - Brain glucose bisphosphatase requires inosine monophosphate. AB - Glucose bisphosphate phosphatase has been partially purified from the cytosol of mouse brain. Enzyme activity required Mg2+ and a heat-stable cofactor. The activator was present in boiled extracts of mouse brain mitochondrial-nuclear fraction, of red blood cells, or of rabbit muscle. The chemical properties of the activator are consistent with its identification as inosine monophosphate (IMP), including its mobility in a high pressure liquid chromatography (HPLC) system capable of resolving all of the biologically important mononucleotides. A large number of other biologically important compounds were not effective, including AMP, cAMP, cGMP, and UMP, GMP, purified by HPLC, (50 or 74 microM), gave a rate about 35% of that obtained with IMP (5 microM). The enzyme was separated completely from phosphoglucomutase and significantly from glucose bisphosphate synthase. The products of the reaction are glucose-P and Pi. Fructose bisphosphate at 500 microM inhibited only 40% in the presence of 20 microM glucose bisphosphate. The activation by IMP follows hyperbolic kinetics with an apparent Ka of 5 microM in the presence of 12 microM glucose bisphosphate. The apparent Km of glucose bisphosphate was 10 microM in the presence of 50 microM IMP. There was no inhibition by 5 or 50 microM AMP or ADP. The possible regulatory importance of glucose bisphosphate in carbohydrate metabolism and the significance of the regulation of the phosphatase by the nucleotide are discussed. PMID- 6282818 TI - A multiplicity of opiate receptors and enkephalin neuronal systems. AB - Properties of the opiate receptor, such as its discrimination of agonists and antagonists, and the relationship between receptor localization and central sites of pain perception suggested that this receptor might interact with a normally occurring opiate-like substance. Such opioid peptides have now been described in at least two systems of distinct neurons which occur in areas particularly associated with pain and emotion. Understanding the relationship between the multiple opiate receptors described pharmacologically and those distinguished by binding studies in methionine- or leucine-enkephalin-containing neurons might be useful for the development of new drugs that suppress pain or alter emotion without producing tolerance or withdrawal effects. PMID- 6282820 TI - Identification of ubiquinone-50 in human neutrophils and its role in microbicidal events. PMID- 6282821 TI - The acid phosphatase with optimum pH of 2.5 of Escherichia coli. Physiological and Biochemical study. AB - In Escherichia coli, the physiological conditions governing the expression of an acid phosphatase with an optimum pH of 2.5 were determined. By contrast with most enzymes, the synthesis of this phosphatase was turned off in exponentially growing bacteria and started as soon as cultures entered the stationary phase. A starvation for inorganic phosphate resulted in a premature full induction, while carbon, nitrogen, and sulfur limitations were inefficient. In the presence of nonlimiting amounts of inorganic phosphate, however, the transfer of the culture to anaerobic conditions led to an immediate accumulation of the acid phosphatase. Cyclic AMP exerted a strong negative control on the biosynthesis and of this enzyme for which the integrity of both the cya and the crp gene functions was necessary. The acid phosphatase was purified to apparent homogeneity and behaved as a monomeric protein with a molecular weight of about 45,000. It had predominantly a phosphoanhydride phosphatase activity and preferentially hydrolyzed the gamma-phosphoryl residue of GTP (Km = 0.35 mM) and the 5'-beta phosphoryl residue of ppGpp (Km = 1.8 mM). The corresponding beta-phosphoryl residue of GDP was little hydrolyzed, while CTP, ATP, and UTP were not. The enzyme did not split most phosphomonoesters with the exception of the synthetic substrate p-nitrophenyl phosphate (Km = 2.7 mM), 2,3-bisphosphoglycerate (Km = 5 mM), and fructose 1,6-bisphosphate (Km = 5 mM). It was competitively inhibited by tartaric acid and by sodium fluoride (Ki = 60 microM). In addition, it was sensitive to the inhibitor of the translation elongation factor EF-G fusidic acid, and was also strongly inhibited by the triazine dye Cibacron Blue F3GA (Ki = 0.3 microM), suggesting the existence of a site able to recognize nucleotides. PMID- 6282822 TI - Localization of lipid binding domain(s) on subunit II of beef heart cytochrome c oxidase. AB - Photoactivatable arylazido phospholipids (Bisson, R., and Montecucco, C. (1981) Biochem. J. 193, 757-763) have been used to study the arrangement in the membrane of beef heart cytochrome c oxidase subunit II. Fragmentation of the photocross linked polypeptide showed that sequence 20-98, which contains two long stretches of uncharged amino acid residues (Steffens, G. J., and Buse, G. (1979) Hoppe Seyler's Z. Physiol. Chem. 360, 613-619), is in contact with lipids. Edman degradation of the labeled peptide indicated that His 26, Thr 27 and Met 29 are involved in the interaction with the polar head region of the bilayer. These data have been used to propose a scheme for the folding of subunit II in the mitochondrial membrane. PMID- 6282823 TI - Alterations of glial tumor cell Ca2+ metabolism and Ca2+-dependent cAMP accumulation by phorbol myristate acetate. AB - C6 glial tumor cells exposed to phorbol myristate acetate (PMA) possessed lowered cAMP content, reduced ability to accumulate cAMP in response to norepinephrine or cholera toxin, and a 3-fold increase in the concentration of norepinephrine producing 50% of the maximal rate of cAMP accumulation. Detectable effects on cAMP accumulation occurred within 10 min of exposure to PMA, and prominent effects by 2 h. PMA similarly affected cells pretreated with cycloheximide. In contrast, Ca2+-depleted preparations of control and PMA-treated cells accumulated cAMP identically in response to norepinephrine or cholera toxin. Ca2+ restoration, which increased the rate of cAMP accumulation in control cells severalfold, did not enhance cAMP accumulation in PMA-treated cells. Neither high catecholamine nor high extracellular Ca2+ concentrations reversed the suppression of cAMP accumulation by PMA. Trifluoperazine, which inhibited the Ca2+-dependent component of norepinephrine-stimulated cAMP accumulation in control cells, did not significantly reduce norepinephrine-stimulated cAMP accumulation in PMA treated cells. Cell free preparations of control and PMA-treated cultures did not differ significantly in calmodulin content or in Ca2+-stimulated adenylate cyclase, Ca2+-dependent cAMP phosphodiesterase, and (Ca2+-Mg2+)-ATPase activities. The Ca2+ content, however, of intact cells decreased with time of PMA treatment. Within minutes after exposure to PMA, the ability of Ca2+-depleted cells to take up 45Ca was significantly reduced. Both 45Ca uptake and Ca2+ dependent cAMP accumulation were reduced over the same PMA concentration range. PMID- 6282824 TI - Interaction between 1,25-dihydroxyvitamin D3 receptors and intestinal nuclei. Binding to nuclear constituents in vitro. AB - The molecular action of 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) is thought to involve its localization within the nucleus of target cells, a process mediated by intracellular receptors. This report probes both the association between chick intestinal 1,25(OH)2D3 receptors and purified homologous nuclei and the interaction between this receptor and nucleic acids. 1,25(OH)2D3 receptors bound to purified nuclei in a apparently saturable manner (Kd = 2.2-4.8 X 10(-10) M) under conditions of intermediate ionic strength and constant protein concentration. Nuclear binding was hormone-dependent; whereas receptor-hormone complex (Rs) binds to nuclei under the ionic conditions employed here (greater than 70%), hormone-free (R0) receptors do not bind (less than 10%). Binding was localized to the nuclear chromatin fraction and was extremely sensitive to KCl concentration both in the incubation medium and during postincubation treatment of nuclei. The interaction appeared to be temperature-independent, suggesting the lack of a classic activation event characteristic of most steroid receptors. Partial digestion of intestinal nuclei with DNase I eliminated subsequent receptor binding by greater than 95%, pointing to the involvement of DNA in the binding interaction. In turn, receptors were found to bind to both DNA and RNA, a characteristic independent of receptor aggregation, but sensitive to disruption with increasing ionic strength buffers. Elution of both Rs and R0 from DNA appeared identical (0.28 M KCl), whereas the strength of interaction with RNA was much less (0.12 M KCl). Thus, while there appeared to be a fundamental difference between R0 and Rs, such that only the binding of receptor-hormone complex to nuclei was allowed under the conditions employed here, this characteristic was not observed during DNA binding. Nevertheless, the possibility exists that the in vivo interaction between 1,25(OH)2D3 receptor and nuclei involves DNA and that this nuclear constituent may be the ultimate site of action of this unique sterol hormone. PMID- 6282825 TI - Gene conversion of two functional goat alpha-globin genes preserves only minimal flanking sequences. AB - We have determined the complete nucleotide sequence of the nonallelic adult goat I alpha- and II alpha-globin genes and, as is the case with the duplicated human alpha-to each other. Such high homology (99%) has most likely been preserved via a gene conversion mechanism. The conversion unit in goats is only about 9000 base pairs in length, and contained within this short region are all the known signals required for accurate and efficient transcription, with the CCAAT box adjacent to the 5'-boundary of the conversion unit and the poly(A) addition site adjacent to the 3' end. This conversion unit is also flanked by a 23-base-pair direct repeat "boundary sequence," vestiges of which are also observable in the human and mouse alpha-globin genes and pseudogenes. These direct repeats imply that a transposition-like event may have been responsible for the insertion of an ancestral alpha-like sequence into a new chromosomal locus, and that this insertion event and subsequent gene duplication may have predated the mammalian radiation. PMID- 6282827 TI - Development of insulin responsiveness of the glucose transporter and the (Na+,K+) adenosine triphosphatase during in vitro adipocyte differentiation. AB - The development of insulin responsiveness of the transport systems for glucose (2 deoxyglucose) and potassium (Rb+) was compared during in vitro adipocyte differentiation of 3T3-L1 cells. Growing cells exhibited minimal increases in 2 deoxyglucose and Rb+ transport rates in response to insulin. Low levels of insulin stimulation of both transport processes became apparent when cultures attained confluence, and these levels were maintained in uninduced cells which retained their fibroblast-like morphology. Following a 48-h induction treatment with methylisobutylxanthine and dexamethasone, a dramatic and simultaneous increase in insulin sensitivity of both deoxyglucose and Rb+ uptake was observed. In fully differentiated 3T3-L1 adipocytes, insulin caused a 6-10-fold increase above the basal rate for deoxyglucose uptake and a 1.50-1.60-fold increase of Rb+ uptake. The insulin dose response relationships were identical for both deoxyglucose uptake and Rb+ uptake and half-maximal stimulation occurred at insulin concentrations of 2-3 nM in 3T3-L1 fibroblasts, 550 pM in 3T3-L1 adipocytes, and 100 pM in mature rat adipocytes. Basal rates of deoxyglucose and Rb+ uptakes were 2-9-fold higher in growing cells than in confluent cells, and fatty cells exhibited lower transport rates relative to nonfatty cells. The number of active Na+ pumps on the cell surface was determined by quantitation of the covalent phosphorylated intermediate of the (Na+,K+)-ATPase. Plasma membranes of growing cells contained a larger number of (Na+,K+)-ATPase specific phosphorylation sites and higher (Na+,K+)-ATPase activity than those of confluent cells. Although (Na+,K+)-ATPase and Rb+ transport activities were greater in uninduced cells than in induced cells, both cultures exhibited the same number of phosphorylation sites, implying that the Na+ pump in 3T3-L1 adipocytes was operating at a reduced efficiency. PMID- 6282826 TI - Prolactin receptors in cultured rat mammary tumor cells. Unmasking of cell surface receptors by energy depletion. PMID- 6282828 TI - (Na+,K+)-ATPase kinetics within the intact renal cell. The role of oxidative metabolism. AB - The kinetics of oxygen consumption and (Na+,K+)-ATPase-mediated K+ transport was examined by reintroducing K+ into a K+-depleted suspension of renal tubules. In the presence of the substrates glucose, lactate, and alanine, a K+/O2 ratio of 10.4 +/- 0.2 was obtained, and the apparent K1/2 for K+ transport with respect to external K+ concentration was 0.9 mM. Supplementation of the substrates with the short chain fatty acid, butyric, had a 3-fold effect on the kinetic parameters examined: 1) the quantity of (Na+,K+)-ATPase-mediated ion transport per oxygen consumed fell by 17 +/- 2%; 2) the maximum rate of K+ transport increased by nearly 50%; and 3) the apparent K1/2 for transport with respect to external K+ concentration rose to 1.5 mM. These results indicate that despite decreasing the quantity of ATP produced per oxygen consumed, short chain fatty acids are able to increase the overall production of ATP during periods of high metabolic demand. The coupling between the two major metabolic processes of the renal cell, (Na+,K+)-ATPase-mediated ion transport and mitochondrial oxidative phosphorylation, is addressed in the context of these findings. PMID- 6282829 TI - Purification and characterization of a wheat germ protein kinase. AB - A cyclic AMP-independent protein kinase has been purified from wheat germ extracts. The enzyme catalyzes the phosphorylation of casein and phosvitin but not protamine, histone, or bovine serum albumin. However, the best substrate for the kinase appears to be that of an endogenous wheat germ protein. The kinase can utilize both ATP and GTP as phosphoryl donors. A molecular weight of 36,000 38,000 had been estimated for the kinase by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and by glycerol density gradient centrifugation and gel filtration in the presence of 0.5 M KCl. In the presence of low salt, however, the molecular weight of the kinase appears to double. In isoelectrofocusing, the kinase exhibits a pI of about 6.5. The activity of the kinase is strongly inhibited by spermine and heparin. Spermidine is slightly stimulatory at low concentrations but inhibitory at high concentrations. High concentrations of putrescine also inhibit the kinase activity, but not to the extent observed with the other polyamines. Both spermine and spermidine appear to enhance the kinase activity at low Mg2+ concentrations. The result suggests that these polyamines could partially replace Mg2+ for kinase activity. PMID- 6282830 TI - Biochemical, biological, and immunological properties of chemically deglycosylated human choriogonadotropin. AB - A chemical method of deglycosylation of human choriogonadotropin (hCG) was used to assess the role of carbohydrate moiety in the maintenance of quaternary structure and functional parameters such as receptor binding, immunological activity, and in vitro biological response. Treatment of purified hCG with anhydrous HF at 0 degrees C for 60 min was effective in removing more than 75% of the carbohydrate moiety. This extent of deglycosylation altered its chromatographic characteristics as revealed by retarded behavior on Sephadex G 100 and failure to be retained on concanavalin A-Sepharose. The electrophoretic heterogeneity present in native hCG was markedly reduced by deglycosylation. The deglycosylated hCG was stable in the lyophilized form and retained its quaternary structure as revealed by the fluorescence probe 8-anilino 1-naphthalene sulfonic acid, receptor binding, and immunological activities. Unlike receptor binding and immunological activities, which were fully retained, the ability of the hormone to stimulate cyclic AMP accumulation in vitro in rat interstitial cells was completely abolished. PMID- 6282831 TI - Basement membrane collagen synthesis by rabbit corneal endothelial cells in culture. Evidence for an alpha chain derived from a larger biosynthetic precursor. AB - Corneal endothelial cells in culture synthesize basement membrane collagen and secrete it into the medium. This collagen sediments faster than interstitial collagen by velocity sedimentation and is disulfide-bonded. After reduction, two biochemically distinct chains can be determined by cyanogen bromide peptide mapping. These chains migrate close to each other and immediately below beta 12(I) components on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Treatment with pepsin gives rise to a major band which still retains interchain disulfide bonds and which will not convert to components with the mobility of interstitial alpha chain by reduction. However, an alpha chain and three minor collagenase-sensitive and pepsin-resistant peptides are generated if the molecule is reduced and alkylated under nondenaturing conditions prior to pepsin treatment. When collagen which accumulates in the media over a long period of time is compared to the newly synthesized molecules, there is an apparent differential resistance to limited pepsin treatment. However, the products which are generated in both cases share electrophoretic identity. PMID- 6282832 TI - Purification of a tyrosine-specific protein kinase from Rous sarcoma virus induced rat tumor. AB - We have identified a tyrosine kinase activity present in tumors which were raised in rats by subcutaneous injection of Rous sarcoma virus-transformed rat cells (SR NRK). This kinase phosphorylates tyrosine on the heavy chain of IgG from tumor bearing rabbit (TBR) sera specific for the src gene product, pp60src. Using TBR IgG phosphorylation as an assay, we have purified this kinase over 7200-fold. The purification procedure involves detergent extraction of tumors followed by sequential column chromatography on hydroxylapatite, DEAE-Sephacel, oligodeoxyadenosine-cellulose, an affinity column prepared from TBR-sera, and Sephacryl S-200. The IgG kinase activity behaves as a molecule of apparent Mr = 54,000 on Sephacryl S-200 molecular sieve chromatography. Analysis of the Sephacryl fractions by SDS-PAGE indicates that a major Coomassie blue-stained band with an apparent Mr = 54,000 (p54), co-elutes with the peak of kinase activity. From 600 g of tumors, approximately 200 micrograms of p54 are obtained. We have four types of evidence which show that p54 is related to pp60src. 1) Purified p54 is capable of undergoing endogenous phosphorylation in the presence of [gamma-32P]ATP producing a 32P-labeled pp54 polypeptide which is specifically immunoprecipitated by TBR-sera and contains only phosphotyrosine. 2) Purified p54 competes with 32P-labeled pp60src for binding to TBR-IgG, indicating a degree of purification over starting material which agrees very well with the results obtained by the IgG kinase assay. 3) V8 protease digestion of pp60src and p54 suggests that they share a common 26,000 fragment. 4) Antibodies to partially purified p54 specifically precipitate pp60src from Rous sarcoma virus-transformed chicken cells. PMID- 6282833 TI - Properties of the src kinase purified from Rous sarcoma virus-induced rat tumors. AB - We recently described a method for the purification of a protein kinase related to pp60src from Rous sarcoma virus-induced rat tumors (Blithe, D. L., Richert, N. D., and Pastan, I. H. (1982) J. Biol. Chem. 257, 7135-7142). In this report, we describe some of the properties of the 7200-fold purified enzyme. The purified kinase phosphorylates casein, vinculin, actin, and histone H2B, but not bovine serum albumin or ovalbumin. Protein substrates are phosphorylated exclusively on tyrosine residues. Casein was used as a substrate for more detailed analysis. The phosphorylation reaction proceeds at a linear rate for at least 40 min at 22 degrees C. Maximum enzyme activity occurs at pH 6.5 to pH 6.8 and requires the presence of either Mg2+ (5 to 10 mM) or Mn2+ (1 to 5 mM). The Km for ATP is 30 microM and the Vmax 0.03 mumol/min/mg using 0.4 mg/ml of casein as a substrate. The enzyme utilizes ATP or dATP, but not GTP as a phosphate donor in the reaction. The enzyme is inhibited by adenosine and deoxyadenosine and by their corresponding mono- and diphosphates. No inhibition of enzyme activity is observed with adenine, GTP, UTP, TTP, or CTP. The enzyme is very sensitive to increased ionic strength. Addition of 0.1 M KCl, 0.1 M NaCl, 50 mM KPO4, or 50 mM NaPO4 inhibited casein phosphorylation by 90 to 95%. Analysis of the products of the phosphorylation reaction by thin layer chromatography revealed that the src kinase phosphorylates glycerol in addition to casein or the enzyme itself. PMID- 6282834 TI - Targeted deletion of a yeast enolase structural gene. Identification and isolation of yeast enolase isozymes. AB - Yeast contain two nontandemly repeated enolase structural genes which have been isolated on bacterial plasmids designated peno46 and peno8 (Holland, M. J., Holland, J. P., Thill, G. P., and Jackson, K. A. (1981) J. Biol. Chem. 256, 1385 1395). In order to study the expression of the enolase genes in vivo, the resident enolase gene in a wild type yeast strain corresponding to the gene isolated on peno46 was replaced with a deletion, constructed in vitro, which lacks 90% of the enolase coding sequences. Three catalytically active enolases are resolved differ DEAE-Sephadex chromatography of wild type cellular extracts. As expected, a single form of enolase was resolved from extracts of the mutant cell. Immunological and electrophoretic analyses of the multiple forms of enolase confirm that two enolase genes are expressed in wild type cells and that isozymes are formed in the cell by random assortment of the two polypeptides into three active enolase dimers. The yeast enolase loci have been designated ENO1 and ENO2. The deletion mutant lacks the enolase 1 polypeptide confirming that this polypeptide is encoded by the gene isolated on peno46. The intracellular steady state concentrations of the two polypeptides are dependent on the carbon source used to propagate the cells. Log phase cells grown on glucose contain 20-fold more enolase 2 polypeptide than enolase 1 polypeptide, whereas cells grown on ethanol or glycerol plus lactate contain similar amounts of the two polypeptides. The 20-fold higher than in cells grown on the nonfermentable carbon sources. In vitro translation of total cellular RNA suggests that the steady state concentrations of the two enolase mRNAs in cells grown on different carbon sources are proportional to the steady state concentrations of the respective enolase polypeptides. PMID- 6282835 TI - RNA capping by HeLa cell RNA guanylyltransferase. Characterization of a covalent protein-guanylate intermediate. AB - RNA capping by partially purified HeLa cell GTP:RNA guanylyltransferase has been shown to occur in the following sequence of two partial reactions involving a covalent protein-guanylate intermediate: (i) E(P68) + GTP in equilibrium E(P68 GMP) + PPi (ii) E(P68-GMP) + ppRNA in equilibrium GpppRNA + E(P68) Initially, the enzyme reacts with GTP in the absence of an RNA cap acceptor to form a covalent protein-guanylate complex. This complex consists of a GMP residue linked via a phosphoamide bond to a Mr = 68,000 protein. The enzyme then transfers the guanylate residue from the Mr = 68,000 polypeptide to the 5' end of diphosphate terminated poly(a) to yield the capped derivative GpppA(pA)n. Both partial reactions have been shown to be reversible. In the reverse of Reaction i, E(P68- GMP) reacts with PPi to regenerate GTP. In the reverse of Reaction ii, the enzyme catalyzes the transfer of the 5'-GMP from capped RNA to the Mr = 68,000 protein to form protein-guanylate complex. A divalent cation is required for both partial reactions. The Mr = 68,000 protein is presumed to be a subunit of the HeLa guanylyltransferase. This interpretation is consistent with the sedimentation coefficient of 4.2 S of the native enzyme. Preliminary studies of RNA guanylyltransferase from mouse myeloma tumors suggest a similar mechanism of transguanylylation involving a Mr = 68,000 protein-guanylate complex. These data, in conjunction with previous studies of vaccinia virus guanylyltransferase (Shuman, S., and Hurwitz, J. (1981) Proc. Natl. Acad. Sci. U. S. A. 78, 187-191) suggests that covalent GMP-enzyme intermediates may be a general feature of the RNA capping reaction. PMID- 6282836 TI - Transcription of the chicken alpha 2 (Type I) collagen gene by homologous cell free extracts. AB - We have used two methods to detect specific transcription of the chicken alpha 2 (type I) collagen gene in cell-free extracts derived from Rous sarcoma virus transformed chicken embryo fibroblasts. The first method is a modification of the S1 nuclease mapping procedure which utilizes a DNA probe labeled with 32P at the 5' end of the HindIII linker originally used to clone the collagen promoter region into PBR322. The probe distinguishes newly made, specific RNA from endogenous RNA and nonspecific transcripts. Using this procedure we have found that chicken whole cell extracts support accurate initiation of transcription of the chicken alpha 2 (type I) collagen DNA template. Addition of either creatine phosphate, GTP, or UTP to concentrations of approximately 3 to 5 mM was found to stimulate RNA polymerase II transcription by 5- to 10-fold. The second method employs an avian myeloblastosis virus reverse transcriptase-catalyzed primary extension procedure, rendered in vitro-specific by use of a pBR322 fragment as primer. These two techniques should be useful for analyzing specific transcription in other types of cell-free extracts. PMID- 6282837 TI - Maitotoxin, a Ca2+ channel activator candidate. AB - Effects of maitotoxin, the most potent marine toxin, were studied using a rat pheochromocytoma cell line, PC12h. A low concentration (10(-8) g/ml) of maitotoxin induced a profound increase in CA2+ influx into PC12h cells and the Ca2+-dependent release of [3H]norepinephrine from them. The effects of maitotoxin were not affected by treatment with tetrodotoxin (10(-6) M) and were observed even in the absence of external Na+. Furthermore, these effects were markedly inhibited or abolished by treatment with verapamil (30-300 microM), Mn2+ (5 mM), or tetracaine (1 mM). These results suggest that maitotoxin activates the voltage dependent calcium channels of PC12h cells. PMID- 6282838 TI - Phosphotyrosyl-protein phosphatase of TCRC-2 cells. AB - Homogenization of TCRC-2 cells yielded a phosphotyrosyl-protein phosphatase with a specific activity approximately 10-=fold higher in particulate than in soluble fractions. Over 90% of the phosphotyrosyl-protein phosphatase associated with the particles was solubilized with 1.0% Nonidet P-40. Chromatography of the detergent solubilized particulate fraction on either wheat germ lectin-Sepharose or histone Sepharose columns separated two major components of phosphatase activity. One peak (eluted with 200 mM NaCl from histone-Sepharose or with N-acetylglucosamine from the lectin column) contained both phosphotyrosyl- and phosphoseryl-protein phosphatase as well as p-nitrophenyl phosphatase activities. The other peak (eluted with 1.0 M NaCl from histone-Sepharose or not bound to the lectin column) contained essentially only phosphoseryl-protein phosphatase activity. Various agents (EDTA, p-nitrophenyl phosphate, fluoride) showed considerable differences in their ability to inhibit the two phosphatase fractions; of these, the most potent and selective inhibitor was orthovanadate. At micromolar concentrations, vanadate inhibited the fraction containing phosphotyrosyl-protein phosphatase and failed to inhibit the fraction containing only phosphoseryl-protein phosphatase activity. These data show that the particulate forms of phosphotyrosyl-protein phosphatase and p-nitrophenyl phosphatase represent the activities of very similar or identical proteins. PMID- 6282839 TI - Purification of a myosin phosphatase from bovine aortic smooth muscle. AB - A myosin phosphatase has been purified to homogeneity from bovine aortic smooth muscle. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the enzyme eluted from nondenaturing gels revealed two subunits (Mr = 67,000 and 38,000). Densitometric scans of the subunits indicated a molar ratio of 1:1. Several phosphoproteins were substrates for the phosphatase including histone II-A, isolated 20,000-dalton smooth muscle myosin light chains, phosphorylase a, and smooth muscle myosin. In the presence of 0.25 M NaCl and a substrate concentration of 2 microM, myosin was preferentially dephosphorylated. The specific activity of the phosphatase for myosin at a concentration of 10 microM was found to be 5 mumol/mg/min. The phosphatase required Mn2+ or Co2+ ions for activity. Mg2+, Ca2+, or Mg-ATP would not substitute for Mn2+ or Co2+ at equimolar concentrations. This phosphatase may play an important role in regulating actin-myosin interaction in smooth muscle by serving to dephosphorylate myosin. PMID- 6282840 TI - Nucleotide sequences of switch regions of immunoglobulin C epsilon and C gamma genes and their comparison. AB - Immunoglobulin class switch involves a unique recombination event that takes place at the switch (S) region which is located 5' to each constant region (C) gene of the heavy (H) chain. For example, differentiation of the B lymphocyte from a mu-chain producer to an epsilon-chain producer is mediated by the switch recombination between the S mu and S epsilon regions. In order to elucidate the molecular mechanism for the switch recombination, we have determined nucleotide sequences surrounding the class switch recombination sites of the C epsilon and C gamma 3 genes and those in the 5' flanking regions of the C gamma 2a and C delta genes. The results indicate that the 5' flanking regions of all the CH genes except for the C delta gene contain the S regions which comprise tandem repetition of short unit sequences in agreement with the previous analyses of the S gamma 1, S gamma 2b, S mu, and S alpha regions. Comparison of the nucleotide sequences of all the S regions revealed that length as well as nucleotide sequences of the S regions vary among different classes of the CH gene, but they share short common sequences, (G)AGCT and TGGG(G). The nucleotide sequence of the S mu region is homologous to those of the other S regions in the decreasing order of the S epsilon, S alpha, S gamma 3, and (S gamma 1, S gamma 2b, s gamma 2a) regions. We have compared the nucleotide sequences immediately adjacent to the recombination sites of seven rearranged genes and have always fund tetranucleotides TGAG and/or TGGG, except for one case. Such tetranucleotides may constitute a part of the recognition sequence of a putative recombinase. These results provide further support for our previous proposal that the switch recombination may be facilitated by short common sequences dispersed in all the S regions. PMID- 6282841 TI - Effects of angiotensin II on phosphatidylinositol and polyphosphoinositide turnover in rat kidney. Mechanism of prostaglandin release. AB - Prostaglandin (PG) release from rat inner medullary tissue has been shown to be stimulated by angiotensin II, bradykinin, and arginine vasopressin. PG release from inner medullary has also been demonstrated to be a Ca2+-dependent process. We performed the following studies in an attempt to determine the mechanism by which angiotensin II stimulates PG release and to identify the lipid source serving as the donor for arachidonic acid (AA) in the Ca2+-dependent reaction. After 5 min of incubation, slices of inner medulla prelabeled with [14C]AA released 2.0-fold as much radiolabel into the media in the presence of Ca2+ as in the absence of Ca2+. After 30 min of incubation, the neutral lipids lost 6.1% of their [14C]AA label, phosphatidylethanolamine lost 12.5%, phosphatidylcholine 13.3%, and phosphatidylinotisol (P[I) 27%. The divalent ionophore A23187 (5 microM) increased 3.7-fold the formation of immunoassayable prostaglandin E2 at 30 min in the presence of Ca2+. Angiotensin II increased immunoassayable PGE2 formation 1.3-fold at 2 min and 1.5- to 1.8-fold by 30 min. In addition, angiotensin II rapidly increased the incorporation of [32P]orthophosphate to significantly higher values into PI, phosphatidic acid, diphosphoinositol, and triphosphoinositol by 30 s, returning to control values by 2 min of incubation. The data suggest that PI may be a major source of arachidonic acid in the Ca2+ dependent release of PG, that angiotensin II stimulates a smaller or different pool of AA release than the divalent ionophore, and the angiotensin II stimulation of PG is a Ca2+-mediated process associated with increased "phosphatidylinositol-polyphosphoinositide" turnover in the rat inner medulla. PMID- 6282842 TI - Evidence for the organization of the transmembrane segments of (Na,K)-ATPase based on labeling lipid-embedded and surface domains of the alpha-subunit. AB - The purpose of this work has been to examine the organization of the intramembranous portion of the alpha-subunit of membrane-bound (Na,K)-ATPase. Covalent labeling of the alpha-subunit and its tryptic fragments from within the lipid bilayer with [125I]iodonaphthylazide was combined with covalent labeling with 32P from [gamma-33P]ATP at the cytoplasmic surface and with [3H]N-(ouabain) N'-(2-nitro-4-azidophenyl)ethylenediamine from the extra cellular surface. In control experiments using extensive proteolysis and reduced glutathione, it is confirmed that iodonaphthylazide labels segments of the protein within the lipid bilayer. The labeled segments of the alpha-subunit, produced by extensive proteolysis, are selectively extracted by organic solvents. Both at a low and at a high concentration of iodonaphthylazide, about 50% of label added to the medium is covalently attached to protein and lipid. At the low iodonaphthylazide concentration, the NH2-terminal Mr = 46,000 (46K) fragment of the alpha-subunit is preferentially labeled, while at the higher concentration of the 46K fragment, the 78K fragment, and the COOH-terminal 58K fragment are labeled. 32P from [gamma 32P]ATP is incorporated into the 46K fragment while [3H]N-(ouabain)-N'-(2-nitro-4 azido-phenyl)ethylenediamine from the extracellular surface labels all the major fragments, 78K, 58K, and 46K. The data provide evidence for a model of the path of the polypeptide chain with multiple traverses of the alpha-subunit across the bilayer and the NH2-terminal and three trypsin-sensitive bonds exposed at the cytoplasma surface. PMID- 6282843 TI - Binding of calcium ions to the isolated asialo-glycoprotein receptor. Implications for receptor function in suspended hepatocytes. AB - The binding of calcium ions by the isolated asialoglycoprotein receptor of hepatocytes and the inter-relationship between the calcium ion concentration and receptor function have been studied. The isolated receptor binds calcium ions only in the presence of asialoglycoprotein. The asialo-glycoprotein receptor complex binds 4 calcium ions; the binding exhibits marked positive cooperativity, and the association constant at half-saturation of the binding sites was of the order of 10*5) M-1 as determined from a Hill plot. The isolated receptor was almost saturated at a calcium ion concentration of 0.1 mM. The binding capacity of isolated hepatocytes for asialo-glycoproteins increased, however, even when the calcium concentration was increased above this level. This may be explained by the exposure of increasing numbers of functional receptors on the surface of the cell with increasing membrane potential, and this explanation is supported by analogous observations in the presence of 5 mM La3+. PMID- 6282844 TI - 1,25-Dihydroxyvitamin D receptors in an established bone cell line. Correlation with biochemical responses. AB - A stable cell line derived from mouse bone (cell line MMB-1) has been used for studies of the cellular receptor for 1,25-dihydroxyvitamin D3 in osteoblasts. Previous studies have demonstrated that collagen synthesis in the MMB-1 cell line is specifically inhibited by 1,25-dihydroxyvitamin D3 as well as by other bone regulating hormones. Incubation of cell homogenates with [3H]1,25 dihydroxyvitamin D3 indicated the presence of a specific receptor which was located primarily in the chromatin fraction. Optimum conditions for the receptor assay required the inclusion of 500 kallikrein-inactivating units of Trasylol/ml and 10 mM NaMoO4. Under these conditions the receptors were stable for 2 h at 23 degrees C and for 24 h at 4 degrees C. Cellular content of receptors was dependent upon the state of confluency of the cells: fully confluent cells contained minimal concentrations of receptors. In cultures of 70-80% confluency, the 1,25-dihydroxyvitamin D3 receptors demonstrated linear Scatchard plots with Kd = 0.4 nM. Peak receptor activity was found at 3.7 S in linear sucrose gradient fractions of cell homogenates. The synthesis of collagen by MMB-1 cells was inhibited by 1,25-dihydroxyvitamin D3 in direct proportion to the concentration of cellular receptors at varying levels of culture confluence. The data indicate that MMB-1 cells contain cytoplasmic/nuclear receptors for 1,25-dihydroxyvitamin D3 which are similar to the receptors found in other target tissues for this hormone and suggest that these receptors are mediators of the effects of 1,25 dihydroxyvitamin D3 on collagen synthesis. PMID- 6282845 TI - Hepatic pyruvate kinase. Quantitative measurements of phosphorylation in vitro and in the isolated rat hepatocyte. PMID- 6282847 TI - Rapid changes in the concentration of phosphoenolpyruvate carboxykinase mRNA in rat liver and kidney. Effects of insulin and cyclic AMP. AB - Starvation and diabetes both caused a marked increase in the concentration of hepatic phosphoenolpyruvate caroboxykinase mRNA while the administration of insulin to diabetic rats or refeeding glucose to starved animals caused a marked reduction in the levels of enzyme mRNA as measured by hybridization using a cDNA probe.l The Administration of dibutyryl cAMP to a starved-refed cat caused an 8 fold induction of phosphoenolpyruvate carboxykinase mRNA in 1 h. Triamcinolone plus acidosis induced the levels of enzyme mRNA in kidney 3-fold within 6 h, however, starvation for 24h had only marginal effects. In all of the above conditions, the levels of phosphoenolpyruvate carboxykinase mRNA measured by hybridization assay agreed well with the relative levels of translatable mRNA for the enzyme. The half-time of phosphoenolpyruvate carboxykinase mRNA, determined after the administration of either alpha-amanitin or cordycepin to starved animals, was approximately 40 min. However, cycloheximide either alone or together with cordycepin, not only prevented the decrease in phosphoenolpyruvate carboxykinase mRNA sequence abundance, but induced it 2-fold. Cycloheximide itself, when injected into 21-day fetal rats in utero caused an induction of enzyme mRNA equal to that noted when dibutyryl cAMP was administered. The mRNA for phosphoenolpyruvate carboxykinase is approximately 2.8 kb in length, but nuclei from the livers of diabetic rats contain a number of putative precursor RNA species for the enzyme, up to 6.5 kb in size, all containing a poly(A) tail. Two hours after refeedng glucose to a starved rat, these nuclear RNA species could no longer be detected by hybridization to our cDNA probe. PMID- 6282846 TI - Regulation of rat liver fructose 2,6-bisphosphatase. AB - An enzyme activity that catalyzes the hydrolysis of phosphate from the C-2 position of fructose 2,6-bisphosphate has been detected in rat liver cytoplasm. The S0.5 for fructose 2,6-bisphosphate was about 15 microM and the enzyme was inhibited by fructose 6-phosphate (Ki 40 microM) and activated by Pi (KA 1 mM). Fructose 2,6-bisphosphatase activity was purified to homogeneity by specific elution from phosphocellulose with fructose by specific elution from phosphocellulose with fructose 6-phosphate and had an apparent molecular weight of about 100,000, 6-phosphofructo 2-kinase activity copurified with fructose 2,6 bisphosphatase activity at each step of the purification scheme. Incubation of the purified protein with [gamma-32P]ATP and the catalytic subunit of the cAMP dependent protein kinase resulted in the incorporation of 1 mol of 32P/mol of enzyme subunit (Mr = 50,000). Concomitant with this phosphorylation was an activation of the fructose 2,6-bisphosphatase and an inhibition of the 6 phosphofructo 2-kinase activity. Glucagon addition to isolated hepatocytes also resulted in an inhibition of 6-phosphofructo 2-kinase and activation of fructose 2,6-bisphosphatase measured in cell extracts, suggesting that the hormone regulates the level of fructose 2,6-bisphosphate by affecting both synthesis and degradation of the compound. These findings suggest that this enzyme has both phosphohydrolase and phosphotransferase activities i.e. that it is bifunctional, and that both activities can be regulated by cAMP-dependent phosphorylation. PMID- 6282848 TI - Reactivity and metal-dependent stereospecificity of the phosphorothioate analogs of ATP in the arginine kinase reaction. Structure of the metal-nucleoside triphosphate substrate. PMID- 6282849 TI - Role of exogenous cholesterol in regulation of adrenal steroidogenesis in the rat. AB - Rat steroidogenic tissues take up cholesterol, and it has been suggested that this process plays a regulatory role in steroid hormone synthesis. To provide evidence for this hypothesis, we carried out studies in lipoprotein-deficient rats. Lipoprotein deficiency, achieved by treating male rats with pharmacological amounts of estradiol, led to profound lowering of plasma cholesterol (8 +/- 2 versus 54 +/- 4 mg/dl) and adrenal cholesteryl ester content (113 +/- 57 versus 747 +/- 108 micrograms/organ). Basal serum corticosterone levels were decreased by 50%, and the response to adrenocorticotropic hormone (ACTH) was totally abolished. Injection of high density lipoprotein (HDL) to estradiol-treated animals restored the response of corticosterone to ACTH. Comparable in vitro studies with adrenal cell suspensions obtained from lipoprotein-deficient rats confirmed the in vivo data. Measurement of [14C]acetate incorporation and uptake of both HDL- and low density lipoprotein (LDL)-cholesterol in these adrenal cells showed a progressive increase with the duration of estradiol treatment, and neither of these two phenomena was altered by ACTH. These results provide in vitro and in vivo evidence for the hypothesis that normal adrenal steroidogenesis depends upon cholesterol delivery from plasma. Furthermore, under the conditions studied, ACTH does not stimulate adrenal de novo cholesterol biosynthesis nor the uptake of either HDL- or LDL-cholesterol. PMID- 6282850 TI - Iron binding by horse spleen apoferritin. A vanadyl(IV) EPR spin probe study. PMID- 6282851 TI - Template-prime-dependent turnover of (Sp)-dATP alpha S by T4 DNA polymerase. The stereochemistry of the associated 3' goes to 5'-exonuclease. AB - T4 DNA polymerase converts (Sp)-2'-deoxyadenosine 5'-O-(1-thio[1 18O2]triphosphate) to 2'-deoxyadenosine 5'-O-[18O]-phosphorothioate in the presence of poly(d(A-T).poly(d(A-T)) template-primer. Control experiments involving either omitting the poly(d(A-T)).poly(d(A-T) template-primer or employing the (Rp)-2'-deoxyadenosine 5'-O-(1-thiotriphosphate) diastereomer showed no reaction. It is assumed, therefore, that this conversion as in the P--O case involves incorporation of the thionucleotide into the poly(d(A-T)) followed by hydrolysis resulting from the 3' goes to 5'-exonuclease activity. The 2' deoxyadenosine 5'-O-[18O] phosphorothioate was converted to (Sp)-2' deoxyadenosine 5'-O-(1-thio[1-18O]triphosphate), with no change in the configuration at P alpha by using the coupled adenylate kinase-pyruvate kinase enzyme system. A 31P NMR spectrum of the product showed that the 18O was entirely in the nonbridging position, indicating an overall retention in the net turnover process (i.e. incorporation followed by excision). Since the incorporation process involves an inversion of configuration around the phosphorus (Romaniuk, P. J., and Eckstein, F. (1982) J. Biol. Chem. 257, 7684-7688), it must be inferred that the 3' goes to 5'-exonuclease activity of T4 polymerase proceeds with inversion of configuration at the phosphorus atom, most simply via a direct displacement mechanism. This finding represents the first example of phosphodiester hydrolysis catalyzed by an exonuclease that does not involve a covalent phosphoryl-enzyme intermediate (Knowles, J. R. (1980) Annu. Rev. Biochem. 49, 877-919). PMID- 6282852 TI - Chromatin structure of the chicken beta-globin gene region. Sensitivity to DNase I, micrococcal nuclease, and DNase II. AB - We have examined in some detail the chromatin structure of a 6.2 kilobase pair (kbp) chromosomal region containing the chicken beta-globin gene. The chromatin structure was probed with three nucleases, DNase I, micrococcal nuclease, and DNase II, and the rate of digestion of specific subfragments of the region was compared with the rate of bulk DNA digestion. We have characterized the rate of digestion of each fragment in terms of a sensitivity factor which measures the sensitivity of a fragment to a particular nuclease relative to bulk DNA. The sensitivity factors were determined by a least squares curve fitting method based on target analysis. In nuclei isolated from 14-day-old chicken embryo red blood cells, the entire 6.2-kbp region shows approximately a 10- to 20-fold increase in sensitivity to DNase I, a 3-fold increased sensitivity to micrococcal nuclease, and a 6-fold increased sensitivity to DNase II. In addition to the adult beta globin gene, this region contains 5' and 3' flanking sequences, the 5' half of the inactive, embryonic globin gene, epsilon, and some repeated sequences. There is no obvious correlation between these genetic elements and the overall chromatin structure as measured by the nuclease sensitivity. This same region shows little or no special sensitivity in nuclei isolated from 14-day-old chicken embryo brain. Furthermore, fragments of the inactive ovalbumin gene show little or no sensitivity in either red blood cells or brain. These results support the conclusion that the entire 6.2-kbp region is largely packaged as active chromatin in 14-day-old chicken embryo red blood cells. PMID- 6282853 TI - The molecular characterization of three transcriptional mutations in the yeast iso-2-cytochrome c gene. AB - Three mutations, each of which causes overproduction of iso-2-cytochrome c, were characterized biochemically. Two, CYP3-4 and CYP3-15, were previously shown to be cis-dominant and map to the CYC7 locus which encodes the iso-2 protein, while the third, cyp1-16, maps to an unliked locus. All three mutations caused dramatically increased levels of transcription of the CYC7 gene, and the CYC7 mRNA in mutant cells was found to be the same size as that in wild type cells. The CYP3-4 mutation was found to be caused by the integration of a transposable element, Tyl, 269 base pairs 5' to the coding sequences. The CYP3-15 mutation was also found to alter the DNA, probably through a deletion or inversion with one endpoint 285 base pairs upstream from the coding sequence. The CYC7 gene in both wild type and mutant cells was not subject to catabolite repression. PMID- 6282854 TI - Poly(ADP-ribosylation) in vitro. Reaction parameters and enzyme mechanism. PMID- 6282855 TI - Action of mung bean nuclease on supercoiled PM2 DNA. AB - Single strand specific mung bean nuclease was used to probe for regions of altered secondary structure in supercoiled PM2 DNA. Supercoiled DNA is cleaved greater than or equal to 10,000 times faster than the relaxed topoisomer. Catalytic quantities of enzyme convert supercoiled DNA to both nicked-circular and unit length linear forms at pH 5 but to predominantly the nicked-circular form near neutral pH. At the elevated enzyme concentrations required to cleave relaxed DNA, unit length linear DNA and smaller fragments are produced from pH 5 to 7. One nick per supercoiled DNA molecule is introduced at pH 6.6. The nicks are repairable by DNA ligase and are not strand-specific. Snake venom phosphodiesterase selectively cleaves the strand opposite the nicks, permitting restriction endonuclease mapping. The nicks occur at three specific sites. Sites at 0.75 and 0.76 map units are cleaved with equal frequency, while a site at 0.82 is cleaved less frequently. The former sites map near one of the eight known early denaturation regions of PM2 DNA, while the latter does not. Since most early denaturation sites are not cleaved, sites other than these dA + dT-rich regions may be the preferred locations of strand unwinding and separation in supercoiled PM2 DNA. PMID- 6282856 TI - Structure of the Syrian hamster ribosomal DNA repeat and identification of homologous and nonhomologous regions shared by human and hamster ribosomal DNAs. AB - We have cloned and partially characterized Bam HI fragments of Syrian hamster DNA containing most of the ribosomal RNA-coding region. Several restriction site polymorphisms within the transcribed portions of the hamster rDNA repeats have been noted. Approximately three-fifths of the repeats contain a Bam HI site upstream of the 18 S coding sequences. Approximately four-fifths of the repeats contain a Bam HI site very close to the 5' end of the 28 S coding sequences. This microheterogeneity has been maintained in the DNA of baby hamster kidney (BHK)-21 cells, a cell line established nearly 20 years ago. R-loop analysis with homologous hamster rRNAs has established the size of the coding regions and the internal transcribed spacer. Heterologous R-loop analyses with cloned hamster rDNAs and human rRNAs reveal several well-defined regions within the 28 S gene where the homology between human and hamster RNAs is greatly reduced. These regions are not detectable in heteroduplexes of hamster and human rDNAs. Sequences encoding the 18 S gene do not exhibit such reduced homology. PMID- 6282857 TI - Purification and polypeptide characterization of complex III from yeast mitochondria. AB - Complex III was isolated and purified from bakers' yeast by ammonium sulfate fractionation and column chromatography on Ultrogel AcA 34. The purified complex contained 7.03 nmol/mg of protein and 4.24 nmol/mg of protein of cytochromes b and c1, respectively. The specific activity of the complex was 17.1 mumol/min/mg of protein, using the decyl analog of coenzyme Q as substrate. Electrophoresis of the purified complex revealed the presence of seven polypeptides with molecular weights ranging from 15,500 to 50,000. Polypeptides having molecular weights lower than 15,000 were not observed, except when the complex was dissociated in the absence of proteolytic inhibitors, suggesting that these low molecular weight species arise as a result of proteolytic digestion of the complex. The isoelectric points of the subunits of complex III and their stoichiometry wee determined. Trypsin and chymotrypsin digestion of the oxidized and reduced forms of the isolated complex suggested that the two high molecular weight core proteins are embedded within the complex and hence are inaccessible to the exogenous proteases, while cytochromes b and c1, the iron-sulfur protein, and the 17,500-dalton subunit are substantially exposed to the surface of the complex. The iron-sulfur protein appears to undergo a conformational change upon reduction of the complex, rendering it less susceptible to trypsin digestion. The core proteins and the iron-sulfur protein were purified, and antibodies against these proteins were raised. Immunoinhibition studies with these antibodies also indicated that the antigenic sites of the core proteins were embedded in the complex. PMID- 6282858 TI - Structure of amplified normal and variant dihydrofolate reductase genes in mouse sarcoma S180 cells. AB - We constructed a gene library from a murine cell line with amplified dihydrofolate reductase (dhfr) genes by inserting random segments of DNA into lambda Ch4A. From this library, the dhfr gene and 30 kilobase pairs of surrounding DNA were cloned, and the restriction map was determined. All of the coding regions were sequenced and show that the gene spans a total of 31 kilobase pairs and has five intervening sequences in the coding portion of the gene. In addition, two classes of variant dhfr genes were found in the amplified line, which were amplified and present at levels of 10 to 30% of the normal dhfr genes. Numerous repeated sequences were located throughout the gene region, some of which share homology with previously defied families of repeats. PMID- 6282859 TI - The isolation of homogeneous leader peptidase from a strain of Escherichia coli which overproduces the enzyme. AB - Leader (signal) peptidases cleave the NH2-terminal leader sequences of newly synthesized secreted and membrane proteins during, or shortly after, they insert across the membrane. We have constructed a plasmid, pPS9, in which the structural gene for Escherichia coli leader peptidase is under transcriptional control of a lambda promoter, PR. pPS9 also codes for a temperature-sensitive lambda repressor, causing repression of expression of the plasmid leader peptidase gene at 30 degrees C and rapid synthesis of this enzyme at 42 degrees C. Under appropriate growth regimens, leader peptidase is overproduced 75-fold. Leader peptidase from this strain has been purified to homogeneity. It consists of a 37,000-dalton polypeptide which co-migrates with enzyme activity on DEAE cellulose and chromatofocusing chromatography. Antibody to this leader peptidase is described. PMID- 6282860 TI - The presence of zinc in the restriction enzyme Eco RI. AB - We have determined that the restriction endonuclease Eco RI contains 1.0 +/- 0.1 eq of zinc/monomeric enzyme. DNA cleavage by Eco RI is inhibited by ortho phenanthroline after preincubation of the enzyme with the chelating agent. A similar inhibition by the nonchelating meta-phenanthroline is not seen. The sensitivity of the inhibition by the neutral ligand ortho-phenanthroline to preincubation is consistent with the tightly bound and inaccessible nature of the metal site. Extensive dialysis against the ortho-phenanthroline inhibitor leads to the release of the bound metal with the concomitant loss of enzyme activity. The tightly bound Zn2+ cation, then, appears to be necessary for enzyme function. The finding of zinc in Eco RI further illustrates the ubiquity of Zn2+ to DNA protein complexes. PMID- 6282861 TI - Selective phosphorylation of the alpha subunit of the sodium channel by cAMP dependent protein kinase. AB - The alpha subunit of the sodium channel purified from rat brain is rapidly and selectively phosphorylated by the catalytic subunit of cAMP-dependent protein kinase to a level of 3 to 4 mol of 32P/mol of saxitoxin-binding activity. The rate of phosphorylation is comparable to that of the synthetic peptide analog of the phosphorylation site of pyruvate kinase, one of the best substrates for cAMP dependent protein kinase. An endogenous cAMP-dependent protein kinase that is present in the partially purified sodium channel preparations also selectively phosphorylates the alpha subunit. The specificity and rapidity of the phosphorylation reaction are consistent with the hypothesis that the alpha subunit is phosphorylated by cAMP-dependent protein kinase in vivo. PMID- 6282862 TI - Targeted point mutation that creates a unique Eco RI site within the signal codons of the beta-lactamase gene without altering enzyme secretion or processing. AB - A method has been developed for constructing site-specific mutations by using a strongly selectable marker on which to "piggy-back" a desired mutation that may be phenotypically silent. Using this approach, a new unique Eco RI restriction site has been generated at the beginning of the signal codons of the beta lactamase gene of the plasmid pBR322. The consequential alteration of the second amino acid of the signal from Ser to Arg has no effect on either the transport or the processing of the beta-lactamase. PMID- 6282863 TI - Complex formation between thrombin and thrombomodulin inhibits both thrombin catalyzed fibrin formation and factor V activation. AB - Protein C is activated rapidly when thrombin binds to a specific cell surface cofactor protein, thrombomodulin. Studies were initiated to determine the influence of thrombin-thrombomodulin complex formation on the substrate specificity of thrombin. When thrombin binds to thrombomodulin, the resultant complex retains less than 1% of the fibrinogen clotting activity of free thrombin. Permanent alteration of the thrombin molecule is not involved since full clotting activity is regenerated by incubation of the complex with excess diisopropyl phosphothrombin. Unlike the activation of protein C by the thrombin thrombomodulin complex which is dependent on Ca2+, inhibition of fibrinogen clotting activity is not dependent on the presence of divalent metal ions. Formation of the thrombin-thrombomodulin complex also inhibits thrombin activation of factor V. Despite these changes in macromolecular substate specificity, no significant change in the hydrolysis of the synthetic substrates p-tosyl-L-arginine methyl ester and N alpha-benzoyl-L-arginine ethyl ester is detected upon formation of the thrombin-thrombomodulin complex. Formation of this complex results in a slight increase in the Km (from 9.0 +/- 0.4 to 10.2 +/- 0.6 microM) and Vmax (from 230 +/- 10 to 270 +/- 10 mol/s/mol of thrombin) for the specific thrombin substrate H-D-Phe-Pip-Arg-p-nitroanilide. These studies suggest that thrombomodulin has two distinct anticoagulant functions: 1) to inhibit the ability of thrombin to clot fibrinogen and to activate factor V; and 2) to accelerate the formation of the anticoagulant, activated protein C. PMID- 6282864 TI - The detection of two electron paramagnetic resonance radical signals associated with chloroperoxidase compound I. AB - The electron paramagnetic resonance spectra of chloroperoxidase Compound I and native enzyme are compared. Upon the formation of Compound I, the g = 2.62, 2.26, and 1.82 signals associated with native enzyme disappear and are replaced by two new EPR signals, a sharp signal at g = 2.008 and a broad signal at g = 1.73. The g = 2.008 signal accounts for only 2% of the theoretical spins while the broad signal at g = 1.73 accounts for 60 to 70% of the theoretical spins in Compound I. The g = 1.73 broad signal is reminiscent of the broad EPR signal associated with horseradish peroxidase Compound I. however, the chloroperoxidase Compound I signal has a significantly different g value. The results suggest that the g = 1.73 signal represents a porphyrin pi cation radical which has a stronger coupling to the heme ferryl iron than is the case with horseradish peroxidase Compound I. PMID- 6282865 TI - Differences between male and female rats in the regulation of hepatic glycogenolysis. The relative role of calcium and cAMP in phosphorylase activation by catecholamines. AB - The relative importance of alpha- and beta-adrenergic pathways and of their respective intracellular mediators, calcium and cAMP, in the stimulation of phosphorylase alpha induced by catecholamines was studied in hepatocytes isolated from mature male and female rats. The fractional efflux of 45Ca was used as an index of intracellular calcium activity. Our results show that, in females: 1) the activation of phosphorylase alpha induced by 10(-8) to 10(-5) M epinephrine correlates with a rise in cellular cAMP as well as with an increase in 45Ca fractional efflux, 2) both alpha- and beta-agonists stimulate phosphorylase, 3) neither alpha- nor beta-antagonists effectively block the rise in phosphorylase caused by epinephrine, and 4) propranolol suppresses the rise in cAMP while phenoxybenzamine blocks the rise in calcium efflux. On the other hand, we found that in the male: 1) phosphorylase alpha activity is exclusively correlated with a rise in fractional calcium efflux, 2) epinephrine (10(-8) to 10(-7) M) does not increase cAMP and it causes a greater rise in calcium efflux than in the female at all concentrations, 3) phenylephrine increases calcium efflux and phosphorylase activity without affecting cAMP, 4) phenoxybenzamine totally blocks epinephrine action, and 5) beta-agonists and beta-antagonists are without effects. We conclude that, in females, epinephrine utilizes both alpha- and beta adrenergic pathways which activate phosphorylase by calcium or cAMP, respectively, while, in adult male rats, epinephrine increases phosphorylase alpha activity by an alpha-mediated, calcium-dependent and cAMP-independent pathway. PMID- 6282866 TI - Receptor-dependent and receptor-independent degradation of low density lipoprotein in normal rabbits and in receptor-deficient mutant rabbits. AB - Low density lipoprotein (LDL) catabolism was studied using WHHL rabbits, an inbred strain deficient in LDL receptor activity and, thus, an animal model for homozygous familial hypercholesterolemia. WHHL and normal rabbits were injected with [14C]sucrose-LDL and the tissue sites of LDL degradation were determined 24 h later. On degradation of [14C]sucrose-LDL, the [14C]sucrose ligand remains trapped within tissues as a cumulative measure of degradation. The fractional catabolic rate of [14C]sucrose-LDL in Watanabe heritable hyperlipidemic (WHHL) rabbits was reduced (0.024 +/- 0.010 versus 0.063 +/- 0.026 h-1) but, by virtue of the increased plasma pool, total LDL flux was increased (33.5 +/- 9.6 versus 10.6 +/- 4.4 mg of LDL protein/kg/day). Liver was the predominant site of catabolism in both WHHL and normal rabbits (52.7 +/- 6.9 and 56.6 +/- 6.2% of total degradation). About 90% of hepatic catabolism was attributable to parenchymal cells in both cases. Thus, Kupffer cells, a major component of the reticuloendothelial system, do not play a major role in LDL catabolism in WHHL rabbits. Despite receptor deficiency, the relative contribution of various tissues to overall LDL degradation was not greatly altered and the absolute rate of delivery of LDL to all tissues was increased with the exception of the adrenal. Thus, there was no evidence that the increased degradation occurred in any special subset of "scavenger" cells. Nevertheless, local scavenger cell uptake may be critically important, especially in atherogenesis. If it is assumed that receptor-independent degradation occurs at the same rate in the tissues of WHHL and normal rabbits and that catabolism in the absence of receptors is a linear function of concentration, then one can estimate the fraction of uptake in normal tissues mediated by receptors. The difference in the fraction of the plasma LDL pool cleared per unit of time in normal and WHHL rabbits would reflect the contribution of receptors to fractional clearance. By this calculation, receptor-mediated degradation in normal rabbits was 62% overall, 63% in liver, 92% in adrenal, and 83% in gut. PMID- 6282868 TI - Studies on the mechanism of interaction of fructose 2,6-bisphosphate with fructose-1,6-bisphosphatase. PMID- 6282867 TI - Tissue sites of degradation of native and reductively methylated [14C]sucrose labeled low density lipoprotein in rats. Contribution of receptor-dependent and receptor-independent pathways. AB - Low density lipoprotein (LDL) is catabolized by both receptor-dependent and receptor-independent pathways; methylated LDL (MeLDL) is catabolized only by receptor-independent mechanisms. Rats were injected with either LDL or MeLDL labeled with [14C]sucrose and the tissue sites of degradation were determined 24 h later. On degradation, the 14C-labeled ligand remains trapped intracellularly as a cumulative measure of degradation. The fractional catabolic rate (FCR) of [14C]sucrose-MeLDL was lower than that of [14C]sucrose-LDL (0.056 +/- 0.015 versus 0.118 +/- 0.025 h-1, p less than 0.01). Liver was the predominant site of catabolism of both LDL and MeLDL; more than 85% of catabolism was attributable to parenchymal cells in both cases. The fraction of the plasma LDL pool "cleared" per tissue weight per unit of time was determined for individual tissues. The differences in these rates for LDL and MeLDL are an approximation of receptor mediated uptake of LDL. According to this method, 67.4% of hepatic uptake was attributable to receptors, as was 69.5% of adrenal, 65.4% of ovarian, 52.4% of intestinal, and 44.2% of renal uptake. In other studies, rats were continuously infused with LDL to down-regulate and saturate receptor prior to injection of labeled LDL or MeLDL. Rats infused with LDL exhibited a lower FCR for [14C]sucrose-LDL compared to controls (0.077 versus 0.120 h-1); the FCR for sucrose-MeLDL was unchanged by LDL infusion. The fractional degradation rate of [14C]sucrose-LDL by individual tissues was lowered by LDL infusion in liver, adrenal, ovary, and intestine (41.4, 57.3, 23.1, and 32.4% lower than controls, respectively). The determination of receptor dependency by this independent approach supports the conclusions reached using [14C]sucrose-LDL and [14C]sucrose MeLDL in normolipemic animals. PMID- 6282869 TI - Iron-molybdenum cofactor from nitrogenase. Modified extraction methods as probes for composition. AB - Five modifications of the preparative procedure for isolating iron-molybdenum cofactor (FeMoco) from the molybdenum-iron (MoFe) protein of Azotobacter vinelandii nitrogenase have been developed. This variety of isolation methods has established that no single component of the original isolation protocol, i.e. Tris, Cl-, citrate, HPO4(2-), N,N-dimethylformamide, and N-methylformamide, is essential for the effective isolation and/or structural stability of FeMoco, although any of them may act as ligands to FeMoco when present. The acid-bse status (effective pH) of the extracting solvent is a key adjustable parameter in the isolation procedure. The new procedures produced FeMoco with yields, metal analysis, charge, EPR spectrum, and specific activity (after reconstituting crude extracts from A. vinelandii UW45 mutant cells) essentially identical with FeMoco isolated by the original procedure. After purification, FeMoco apparently contains molybdenum, iron, and sulfide in a 1:7:4 ratio with N-methylformamide as a ligand but no amino acid residues, common sugars, coenzyme A, or lipoic acid. Reaction with o-phenanthroline allows quantitation of both adventitious and FeMoco-associated iron. Correlations of total activity after UW45 reconstitution with molybdenum, total iron, and o-phenanthroline-resistant iron contents show that only the last gives a consistent relationship of 35 +/- 5 nmol of C2H4/min/ng atom of Fe. Both o-phenanthroline and EDTA interact with FeMoco to abolish its EPR signal in reactions reversible by additions of Fe2+ or Zn2+, respectively. These and related reactions point against the presence of an endogenous organic component in FeMoco and toward the presence of exogenous ligands and imply a relatively labile coordination sphere whose nature may be determinable by a systematic investigation. PMID- 6282870 TI - Identification of the fibrinogen receptor on human platelets by photoaffinity labeling. AB - Fibrinogen binding to receptors on stimulated platelets is a prerequisite for platelet aggregation. In order to identify the platelet fibrinogen receptor, we modified fibrinogen with the photoreactive, heterobifunctional cross-linking reagent methyl 4-azidobenzoimidate (MABI). MABI-fibrinogen was fully clottable and able to support platelet aggregation. To photoaffinity label the fibrinogen receptor, gel-filtered human platelets were incubated at 37 degrees C in the dark with 200 micrograms/ml of MABI-fibrinogen, 10 microM ADP, and 0.5 mM calcium. Irradiation of these platelets with ultraviolet light resulted in the incorporation of MABI-fibrinogen into the platelet surface. Incorporation could be prevented by excess native fibrinogen suggesting that MABI-fibrinogen had interacted with the fibrinogen receptor before photolysis. Examination of the irradiated platelets by sodium dodecyl sulfate polyacrylamide gel electrophoresis revealed that the photoactivated MABI-fibrinogen had been incorporated into a 105,000 molecular weight membrane polypeptide that also contained the PlA1 antigen. Thus, this polypeptide has the characteristics of the membrane glycoprotein IIIa. Previous studies have shown that thrombasthenic platelets lack this glycoprotein and fail to bind fibrinogen after stimulation by ADP. Consequently, our data suggest that glycoprotein IIIa constitutes at least one component of the platelet fibrinogen receptor. PMID- 6282872 TI - Properties of particulate and solubilized phosphatidylserine synthase activity from Saccharomyces cerevisiae. Inhibitory effect of choline in the growth medium. AB - When radiolabeled serine is incubated with a particulate fraction from Saccharomyces cerevisiae, radioactivity is incorporated initially into phosphatidylserine and gradually appears in phosphatidylethanolamine. Because decarboxylation of phosphatidylserine is blocked by hydroxylamine, phosphatidylserine synthase can be assayed separately. The yeast phosphatidylserine synthase activity 1) exhibits a divalent cation requirement; 2) is stimulated by exogenous CDP-diolein (apparent Km = 0.17 mM); 3) has an apparent Km = 4 mM for L-serine; 4) has a neutral pH optimum; 5) is inhibited by p-hydroxymercuribenzoate; and 6) is reversible in the presence of 5'-CMP, but not 2'-CMP, 3'-CMP, or 5'-AMP. The phospholipid-synthesizing activity is solubilized with Triton X-100 and the enzymatic parameters have been compared with the particulate form of the enzyme. Detergent extracts catalyze the conversion of exogenous purified [31P]CDP-diglyceride to [32P]phosphatidylserine in the presence of Mn2+ and L-serine. Enzyme preparations from cells grown in the presence of choline, that have reduced phospholipid methylation activity (Waechter, C. J., Steiner, M. R., and Lester, R. L. (1969) J. Biol. Chem. 244, 3419-3422), also have substantially less phosphatidylserine synthase activity compared to identical preparations grown in the absence of choline. When choline, phosphocholine, CDP-choline, and phosphatidylcholine are present in vitro, there is no direct inhibitory effect on phosphatidylserine synthase activity. While the inclusion of choline in the growth medium caused a significant reduction in phosphatidylserine synthase activity, it did not appreciably effect the apparent Km values for L-serine and CDP-diglyceride. These results are consistent with choline-grown cells containing less phosphatidylserine synthase activity because of lower amounts of enzyme present or perhaps less active enzyme due to covalent modification. PMID- 6282871 TI - Correlation of low and high density lipoprotein binding in vivo with rates of lipoprotein degradation in the rat. A comparison of lipoproteins of rat and human origin. AB - These studies were done in the rat to correlate the ability of low and high density lipoproteins of rat (rLDL and rHDL) and human (hLDL and hHDL) origin to bind in vivo to specific tissues with the rates at which these same lipoprotein fractions were cleared from the circulation. The adrenal gland and liver manifested the greatest amounts of rLDL binding in vivo, but activity also was found in spleen, lung, kidney, ovary, and intestine. In contrast, little or no such binding was found utilizing either methyl-rLDL or hLDL. rHDL containing E apoprotein bound to the same group of tissues although in lesser amounts, except in the case of ovary and adrenal gland which bound disproportionately greater amounts of rHDL than rLDL. In keeping with these marked differences in tissue binding, the clearance of rLDL from the plasma equaled 847 +/- 36 microliters/h/100 g of rat while that of methyl-rLDL and hLDL was only 368 +/- 8 and 363 +/- 11 microliters/h/100 g of rat, respectively. When the steady state plasma level of rLDL was raised 2.5-fold, the clearance decreased slightly to 705 +/- 20 microliters/h/100 g of rat. The clearance of hLDL remained constant, however, at about 350 microliters/h/100 g of rat even when the plasma hLDL level was raised to very high values. The clearance of rHDL and hHDL equaled 644 +/- 16 and 408 +/- 13 microliters/h/100 g of rat, respectively, reflecting the more similar rate of binding of rHDL and hHDL to the tissues of the rat. Rates of whole animal sterol synthesis were lowered from 28 mumol/h to 8.8 mumol/h or 13 mumol/h by fasting and cholesterol feeding, respectively, and stimulated to 71 mumol/h by cholestyramine treatment. Under these same conditions, hepatic cholesterol synthesis could be lowered from the normal rate of 15 mumol/h to 4.2 mumol/h and raised to 50 mumol/h. None of these treatments, however, affected the plasma clearance of rLDL and rHDL. In contrast, treatment with ethinyl estradiol increased by 3-fold both the hepatic binding and the whole animal plasma clearance of rLDL. Following resection of approximately two-thirds of the liver under carefully controlled metabolic conditions, there was no change in the rate of hepatic cholesterol synthesis or rLDL binding in the remaining liver, but the clearance of chylomicrons, rLDL, and rHDL diminished by 67%, 26%, and 17%, respectively, suggesting that in the rat the liver was responsible for the degradation of approximately 97%, 39%, and 27%, respectively, of these lipoprotein fractions. PMID- 6282874 TI - Respiration-defective Chinese hamster cell mutants containing low levels of NADH ubiquinone reductase and cytochrome c oxidase. PMID- 6282873 TI - Platelet-derived growth factor. Specific binding to target cells. AB - The binding of the human platelet-derived growth factor (PDGF) to Swiss mouse 3T3 cells have been investigated. The binding is specific and reversible. The dissociation constant is approximately 0.7 x 10(-9) M with approximately 400,000 binding sites/cell. Two forms of PDGF, PDGF I (Mr = 31,000) and PDGF II (Mr = 28,000), previously identified (Deuel, T. F., Huang, J. S., Proffitt, R. T., Baenziger, J. U., Chang, D., and Kennedy, B. B. (1981) J. Biol. Chem. 256, 8896 8899 and Deuel, T. F., Huang. J. S., Proffitt, R. T., Chang, D., and Kennedy, B. B. (1981) J. Supramol. Cell Biochem. 5 (Suppl.), 128) bind equally well to 3T3 cells. Polylysine and histone, but not cytochrome c, partially inhibit the binding of PDGF to 3T3 cells. Protamine sulfate blocks binding in a competitive manner and is capable of displacing PDGF previously bound to the cell surface. EDTA influenced neither the binding of PDGF to the cell surface nor the displacement of cell-bound PDGF. At 37 degrees C, PDGF bound to the cell surface is lost and iodotyrosine is released free into the supernatant, with each process having a t 1/2 of approximately 90 min. The binding activity of the putative PDGF receptor is markedly reduced by previous incubation with PDGF, thereby apparently regulating its activity in a manner similar to epidermal growth factor. PMID- 6282875 TI - Divalent cation-nucleotide complex at the exchangeable nucleotide binding site of tubulin. AB - Tubulin was first treated with alkaline phosphatase-agarose to vacate the exchangeable nucleotide binding site and then tested for manganese binding sites by Mn(II) EPR. Buttlaire et al. ((1980) J. Biol. Chem. 255, 2164-2168) have shown that high affinity manganese binding occurs at a single site normally occupied by magnesium. We report that the number of high affinity manganese binding sites per mol of tubulin depends on the number of occupied exchangeable nucleotide binding sites. Thus, removal of nucleotides results in a loss of high affinity manganese binding sites. The EPR spectra of manganese bound to tubulin and to GTP are found to be qualitatively similar. These data indicate that high affinity manganese binding is the result of the formation of a metal-nucleotide complex at the exchangeable nucleotide binding site. In addition it was found that zinc, cobalt, and magnesium bind with approximately equal affinity to this site whereas calcium binds only weakly. PMID- 6282876 TI - Regulation of diacylglycerol metabolism and arachidonic acid release in human amnionic tissue. PMID- 6282877 TI - Exonuclease II from Saccharomyces cerevisiae. An enzyme which liberates 5' deoxyribomononucleotides from single-stranded DNA by a 5' goes to 3' mode of hydrolysis. AB - A DNase, designated Exonuclease II, has been purified 2,000-fold from whole cell extracts of bakers' yeast. It degrades single-stranded but not double-stranded DNA. The enzyme has a pH optimum around 8 and requires at least 2 mM Mg2+ for activity. It is slightly stimulated by dithiothreitol and inhibited by N ethylmaleimide, suggesting that --SH groups are essential for activity; heparin also inhibits the activity. With 0.1 enzyme unit the Km has been determined to 4 nM of DNA ends and Vmax to 0.52 pmol of liberated nucleotides per min. The Mr is around 120,000. The enzyme does not degrade circular single-stranded M13 DNA, whereas linearized M13 DNA is degraded. The products are 5' deoxyribomononucleotides exclusively. Using 5'-end labeled and 3'-end labeled DNA fragments as substrates, partially degraded DNA is only detectable in the latter case, showing that the exonuclease solely attacks DNA from the 5'-end. This distinguishes Exonuclease II from other exonucleases degrading DNA from the 5' end, since they all either produce a mixture of 5'-mono- and oligonucleotides or 3'-mononucleotides. Analysis of 3'-end labeled fragments after incubation shows that the rate of exonucleolytic cleavage was dependent on the DNA sequence at the 5'-end. PMID- 6282878 TI - Sulfur donor ligand binding to ferric cytochrome P-450-CAM and myoglobin. Ultraviolet-visible absorption, magnetic circular dichroism, and electron paramagnetic resonance spectroscopic investigation of the complexes. AB - The binding of thiol, thiolate, thioether, and disulfide sulfur donor ligands to ferric cytochrome P-450-CAM and myoglobin has been investigated by UV-visible absorption, magnetic circular dichroism (MCD), and EPR spectroscopy. For ferric P 450, the binding of all sulfur donors is competitive with substrate binding. Addition of thiols to P-450 leads to interconvertible thiol or thiolate-bound species depending on the thiol acidity (pKa) and the solution ph; ligation of thiols lowers their pKa by about 4 units. In contrast, only the thiolate-bound form is seen for myoglobin regardless of thiol acidity or solution pH (5.5-11.0), indicating that the heme iron of myoglobin is less electron-rich than that of P 450. Thiolate ligands show much higher affinity (Kd approximately 10(-6) M) for ferric P-450 than do thiols (Kd approximately 10(-3) M). The affinity of thioethers for P-450 (Kd approximately 10(-3) M) is pH-independent (pH 5.5-9.0). The observed disulfide coordination to P-450 represents the first example of disulfide ligation to heme iron; no significant evidence for thioether or disulfide binding to myoglobin is seen. Except for the thiolate complexes, the UV visible and MCD spectral properties of the other sulfur donor . P-450 complexes are similar to, although distinguishable from, those of native P-450. The ferric P-450 . thiolate complexes exhibit MCD spectra resembling that of ferrous P-450 . CO; both also exhibit unique hyperporphyrin (split Soret) UV-visible spectra. The EPR spectra of all P-450 complexes examined display very narrow spread g-values such as are characteristic of native P-450, indicating that the endogenous cysteinate axial ligand is retained upon complex formation. The dissimilarities observed between P-450 and myoglobin in their reactivity toward sulfur donor ligands at least partly reflect the variation in heme iron electron density resulting from their different endogenous axial ligands and may, in turn, help to explain their respective physiological functions of oxygen activation and reversible oxygen binding. PMID- 6282879 TI - An analogue of ubiquinone which inhibits respiration by binding to the iron sulfur protein of the cytochrome b-c1 segment of the mitochondrial respiratory chain. AB - A synthetic quinone, 5-n-undecyl-6-hydroxy-4,7-dioxobenzothiazole (UHDBT), inhibits electron transfer reactions in the cytochrome b-c1 segment of the mitochondrial respiratory chain. Addition of UHDBT to isolated succinate cytochrome c reductase complex has effects on reduction of the cytochromes b and c1 by succinate similar to those which result from removal of the iron-sulfur protein from the b-c1 complex. Thus, UHDBT inhibits reduction of cytochrome c1 by succinate and, if antimycin is added before succinate, UHDBT inhibits reduction of cytochrome b in addition to c1. UHDBT increases the midpoint potential of the iron-sulfur protein of the b-c1 complex from +280 to +350 mV at pH 7.2. The inhibitor also shifts the gx peak in the EPR spectrum of the iron-sulfur protein from g = 1.80 to 1.76 and shifts the gz peak from g = 2.02 to 2.03. It causes only a slight shift in the central gy = 1.90 signal. The efficacy of inhibition of cytochrome c reductase activity of isolated reductase complex by UHDBT appears to depend on the oxidation-reduction poise of some component(s) in the b-c1 complex. Inhibition is decreased and there is an extensive lag in the onset of inhibition under conditions favoring oxidation of the b-c1 complex; inhibition increases and the lag is eliminated under conditions favoring reduction of the b c1 complex. The titer for inhibition of cytochrome c reductase activity of isolated reductase complex is one UHDBT per b-c1 complex. With reductase complex from which the iron-sulfur protein of the b-c1 complex is reversibly resolved, the titer for inhibition is proportional to the amount of iron-sulfur protein reconstituted to the complex. These results suggest that UHDBT inhibits mitochondrial respiration by binding to the iron-sulfur protein of the b-c1 complex, possibly at a site which is otherwise involved in binding ubiquinone, and that this binding is enhanced when the iron-sulfur protein is reduced. PMID- 6282880 TI - The role of superoxide in xanthine oxidase-induced autooxidation of linoleic acid. AB - The effect of hydroxyperoxyoctadecadienoic acid, e.g. 13-hydroperoxy-cis,9,trans 11-octadecadienoic acid, on the autooxidation of linoleic acid induced by superoxide radical was examined in a system containing xanthine oxidase, acetaldehyde, and diethylenetriaminepentaacetic acid dissolved in an aqueous phosphate buffer containing 10% ethanol. The superoxide radical is required for autooxidation, as shown by essentially complete inhibition on the addition of superoxide dismutase. Pure linoleic acid was not readily oxidized, but the addition of lipid hydroperoxide markedly stimulated the autooxidation. Addition of 2.8 microM FeCl3 did not produce an increase in the rate of xanthine oxidase induced autooxidation. Spontaneous autooxidation, a process slower than xanthine oxidase-induced autooxidation, was detectable on the time scale of these observations but was slower than the xanthine oxidase-induced autooxidation. Initiation of linoleic acid autooxidation is postulated to result from a reaction between superoxide and lipid hydroperoxide. The nature of this reaction is uncertain, but it does not appear to depend on iron catalysis. PMID- 6282881 TI - Purification and characterization of a rabbit liver calmodulin-dependent protein kinase able to phosphorylate glycogen synthase. PMID- 6282882 TI - Insulin and glucagon regulation of protein phosphorylation in isolated hepatocytes. Persistence, reversibility, and concentration dependence of hormonal effect. Evidence for common phosphorylation sites for both hormones on the Mr = 46,000 protein. PMID- 6282883 TI - Quantitation of estrogen effect on Xenopus laevis albumin mRNA levels by hybridization to cloned albumin cDNA. AB - The isolation and characterization of a Xenopus laevis albumin cDNA clone, pUN18, is described. This clone was identified as a X. laevis albumin cDNA clone by hybridization-mRNA selection-translation and by other techniques. The clone was used in hybridizations to determine the effect of estrogen on albumin synthesis in livers of male X. laevis. We find that albumin mRNA levels remain relatively constant during the massive induction of vitellogenin mRNA synthesis and vitellogenin secretion which follow administration of estrogen to male X. laevis. PMID- 6282884 TI - Structural features of the cell surface receptor for transferrin that is recognized by the monoclonal antibody OKT9. AB - The monoclonal antibody OKT9 reacts specifically with the receptors for transferrin on human cells (Sutherland, D. R., Delia, D., Schneider, C., Newman, R. A., Kemshead, J., and Greaves, M. F. (1981) Proc. Natl. Acad. Sci. U. S. A. 78, 4515-4519; in Leukemia Markers (Knapp, W., ed) pp. 157-160, Academic Press, New York) and has been used to isolate and characterize this receptor. The receptor is a dimeric glycoprotein (Mr = 180,000) composed of two subunits (Mr = 90,000) and has a pI of approximately 5.2. The transferrin receptor appears to be a transmembrane molecule and is phosphorylated, the phosphate group being predominantly on serine residues. The cell surface form of the molecular possesses both complex and high mannose oligosaccharide chains, which do not appear to have a direct role in antibody (OKT9) binding. The molecule can be cleaved into a Mr = 70,000 fragment from the cell surface, suggesting that the major part of the receptor is exposed to the extracellular environment. The released Mr = 70,000 fragments are not disulfide-linked and possess the antibody (OKT9)- and transferrin-binding sites. Cross-linking studies using radiolabeled transferrin suggest that two molecules of transferrin are bound to each Mr = 180,000 receptor dimer. PMID- 6282885 TI - An investigation into the possible physiological roles of vascular alpha 1- and alpha 2-adrenoreceptors. AB - 1 We have investigated the influence of chemical sympathectomy or bilateral adrenal demedullation on vasopressor responses produced by the alpha 1 adrenoreceptor agonist, phenylephrine, and the alpha 2-adrenoreceptor agonist, M 7, in pithed rats to see whether either procedure induced selective supersensitivity to either of these compounds. 5-HT, a tryptaminergic vasopressor agent, was also used to discriminate between nonspecific change in vascular responsiveness and those mediated via alpha-adrenoreceptors. 2 Adrenal demedullation caused a slight reduction in the sensitivity to all agonists. Chemical sympathectomy significantly reduced the vasopressor responses to all doses of phenylephrine, but responses to 5-HT were unchanged. In contrast there was a tendency for responses to M-7 to be greater in sympathectomized rats than in untreated rats. 3 It is concluded that circulating adrenal medullary catecholamines do not physiologically modulate vascular alpha-adrenoreceptor function. The differential effect of chemical sympathectomy on the vasopressor responses to phenylephrine and M-7 might be explained in terms of a small increase in the ratio of vascular alpha 2 : alpha 1-adrenoreceptors, that receive a noradrenergic innervation. PMID- 6282886 TI - A comparison of peripheral pre- and postsynaptic alpha 2-adrenoreceptors using meta-substituted imidazolidines. AB - 1 The cardiac presynaptic activity, as derived from the inhibition of tachycardia to electrical stimulation of the cardiac sympathetic nerve in pithed, normotensive rats of 8 meta-substituted phenyl(imino)imidazolidines (2, 3- and 2,5- derivation) was determined. 2 The agonistic activity of the imidazolidines was measured with respect to vascular alpha 1- and alpha 2-adrenoreceptors. Accordingly, the increase in diastolic pressure of pithed, normotensive rats to intravenous administration of the imidazolidines was evaluated after pretreatment with 5% w/v glucose solution, yohimbine (1 mg/kg), prazosin (0.1 mg/kg) and the combination of both alpha-adrenoreceptor antagonists. 3 With respect to pre- and postsynaptic alpha 2-adrenoreceptors the 2,5-substituted derivatives were generally less active than the corresponding 2,3-substituted isomers. The steric bulk of the substituent at the 5-position of the imidazolidine molecule appeared to govern the activity with respect to cardiac presynaptic alpha 2 adrenoreceptors, whereas no such clear relationship could be derived for the activity on vascular postsynaptic alpha 2-adrenoreceptors. This indicates that there exists a close resemblance between pre- and post-synaptically located alpha 2-adrenoreceptors. However, these receptor sites are not identical. PMID- 6282887 TI - Malignant eccrine acrospiroma. A case report. PMID- 6282888 TI - Orthopaedic management of childhood neuromuscular disease. Part II: peripheral neuropathies, Friedreich's ataxia, and arthrogryposis multiplex congenita. PMID- 6282889 TI - Defective internalization of low density lipoprotein in epidermoid cervical cancer cells. AB - Cells of an epidermoid cancer cell line of human uterine cervix, which possessed a high-affinity, specific receptor for low density lipoprotein (LDL), internalized and degraded [125I]iodo-LDL at a very low rate. In these cells, LDL did not stimulate cholesteryl ester synthesis, nor did it suppress 3-hydroxy-3 methylglutaryl coenzyme A reductase to the same extent as in the control cells. The binding of [125I]iodo-LDL by these cells was not decreased by preincubation of the cells in medium containing LDL. Using ferritin-labeled LDL (F-LDL) and electron microscopy, it was determined that at 4 degrees C the cells bound F-LDL in the same way as other cancer cell lines that did not have a defect in internalization. When these cells were warmed to 37 degrees C the F-LDL remained on the surface, whereas in cells from control cancer cell lines the F-LDL was internalized and was no longer observed on the cell surface. On the basis of the results of these studies it is concluded that cells of this epidermoid cancer cell line have a defective ability to internalize LDL. PMID- 6282890 TI - Cell surface distribution and intracellular fate of asialoglycoproteins: a morphological and biochemical study of isolated rat hepatocytes and monolayer cultures. AB - A combination of biochemistry and morphology was used to demonstrate that more than 95 percent of the isolated rat hepatocytes prepared by collagenase dissociation of rat livers retained the pathway for receptor-mediated endocytosis of asialoglycoproteins (ASGPs). Maximal specific binding of (125)I asialoorosomucoid ((125)I-ASOR) to dissociated hepatocytes at 5 degrees C (at which temperature no internalization occurred) averaged 100,000-400,000 molecules per cell. Binding, uptake, and degredation of (125)I- ASOR at 37 degrees C occurred at a rate of 1 x 10(6) molecules per cell over 2 h. Light and electron microscopic autoradiography (LM- and EM-ARG) of (125)I-ASOR were used to visualize the surface binding sites at 5 degrees C and the intracellular pathway at 37 degrees C. In the EM-ARG experiments, ARG grains corresponding to (125)I ASOR were distributed randomly over the cell surface at 5 degrees C but over time at 37 degrees C were concentrated in the lysosome region. Cytochemical detection of an ASOR-horseradish peroxidase conjugate (ASOR-HRP) at the ultrastructural level revealed that at 5 degrees C this specific ASGP tracer was concentrated in pits at the cell surface as well as diffusely distributed along the rest of the plasma membrane. Such a result indicates that redistribution of ASGP surface receptors had occurred. Because the number of surface binding sites of (125)I ASOR varied among cell preparations, the effect of collagenase on (125)I-ASOR binding was examined. When collagenase-dissociated hepatocytes were re-exposed to collagenase at 37 degrees C, 10-50 percent of control binding was observed. However, by measuring the extent of (125)I-ASOR binding at 5 degrees C in the same cell population before and after collagenase dissociation, little reduction in the number of ASGP surface receptors was found. Therefore, the possibility that the time and temperature of the cell isolations allowed recovery of cell surface receptors following collagenase exposure was tested. Freshly isolated cells, dissociated cells that were re-exposed to collagenase, and perfused livers exposed to collagenase without a Ca(++)-free pre-perfusion, were found to bind 110-240 percent more(125)I-ASOR after 1 h at 37 degrees C that they did at 0 time. This recovery of surface ASGP binding activity occurred in the absence of significant protein synthesis (i.e., basal medium or 1 mM cycloheximide). Suspensions of isolated, unpolarized hepatocytes were placed in monolayer culture for 24 h and confluent cells were demonstrated to reestablish morphologically distinct plasma membrane regions analogous to bile canalicular, lateral, and sinusoidal surfaces in vivo. More than 95 percent of these cells maintained the capacity to bind, internalize, and degrade (125)I-ASOR at levels comparable to those of the freshly isolated population. ASOR-HRP (at 5 degrees C) was specifically bound to all plasma membrane surfaces of repolarized hepatocytes (cultured for 24 h) except those lining bile canalicular-like spaces. Thus, both isolated, unpolarized hepatocytes and cells cultured under conditions that promote morphological reestablishment of polarity maintain the pathway for receptor- mediated endocytosis of ASGPs. PMID- 6282892 TI - Evidence that cAMP-dependent protein kinase and a protein factor are involved in reactivation of triton X-100 models of sea urchin and starfish spermatozoa. AB - A fraction obtained from detergent-extract of sea urchin or starfish spermatozoa using DEAE-cellulose chromatography reactivated Triton X-100 models of the spermatozoa in a cAMP-dependent manner. The DEAE fraction contained cAMP dependent protein kinase with a high level of specific activity. Rabbit muscle inhibitor protein highly specific for cAMP-dependent protein kinases inhibited the ability of the deae fraction to induce reactivation of Triton X-100 models.l This inhibition paralleled inhibition of cAMP-dependent protein kinase activity of the DEAE fraction, suggesting participation of the enzyme in the cAMP dependent reactivation of Triton X-100 models. However, cAMP-dependent protein kinase further purified from the DEAE fraction was incapable of reactivating these models by itself. A protein factor which was separated from the protein kinase in the course of purification of the enzyme was found to also be necessary for the reactivation. When cAMP-dependent protein kinase was pretreated with protein kinase inhibitor before addition of the protein factor, the reactivation of Triton X-100 models was no longer detected. However, after the protein factor had been incubated with cAMP and cAMP-dependent protein kinase, protein kinase inhibitor did not repress reactivation of Triton X-100 models. We propose that the reactivation needs phosphorylation of the protein factor by cAMP-dependent protein kinase. PMID- 6282891 TI - A nonmitogenic analogue of epidermal growth factor induces early responses mediated by epidermal growth factor. AB - Cyanogen bromide-cleaved epidermal growth factor (CNBr-EGF) binds to EGF receptors with reduced affinity compared to the native hormone but fails to induce DNA synthesis. However, at similar receptor occupancy, CNBr-EGF is as potent as EGF in activating early cell responses to the hormone. The phosphorylation of membrane proteins, the stimulation of Na+-K+-ATPase as reflected by the ouabain-sensitive uptake of 86Rb of fibroblasts, changes in the organization of microfilaments and in cell-morphology, and the activation of the enzyme ornithine-decarboxylase are all induced by CNBr-EGF as well as EGF Our results are consistent with the notion that EGF-induced phosphorylation could act as a "second messenger" for the action of various EGF-induced responses such as activation of Na+-K+-ATPase, changes in the cytoskeleton and cell morphology, and the activation of the enzyme ornithine decarboxylase. However, the stimulation of phosphorylation of membrane proteins and other early responses are either not required or necessary but insufficient for the induction of DNA synthesis. Suboptimal concentrations of EGF together with CNBr-EGF stimulate DNA synthesis in human fibroblasts. Other growth factors such as insulin, fibroblast growth factor, and prostaglandin F2 alpha, which potentiate the mitogenic response of EGF, do not effect the response to CNBr-EGF. This suggests that the restoration of the mitogenic properties of CNBr-EGF by suboptimal doses of EGF occurs at the level of EGF receptors or during their processing. PMID- 6282893 TI - Elimination of specific DNA sequences from the somatic nucleus of the ciliate Tetrahymena. AB - Tetrahymena micronuclear DNA fragments have been cloned in the plasmid pBR322. One clone, pTt 2512, has been found to contain the C-C-C-C-A-A hexanucleotide repeat which is also present in the macronuclear rDNA. Further restriction enzyme digestion and hybridization studies suggest that the clone also contains sequences that are not present in the somatic macronucleus. The flanking sequences of the C4A2 repeats in this clone were separated into four restriction fragments, one from one side and three from the other. These fragments were used as probes for Southern hybridization to study the organizations of similar sequences in the macronucleus and micronucleus. All four fragments hybridized to many fragments of restriction enzyme digested micronuclear DNA. However, none of these hybridizations were detected in the macronucleus. Thus, these families of repetitive DNA are completely eliminated from the macronucleus. Further analysis suggested that the four different sequences may be linked at other locations of the genome. Using nullisomic strains of Tetrahymena, it is found that at least one of these sequences is present in more than one chromosome. Studies of various normal and star strains of Tetrahymena suggest that these sequences are stable in the normal micronucleus but are altered drastically in the defective micronuclei of the star strains. Eliminated DNA of similar nature has also been found in at least five other randomly selected clones of micronuclear DNA and may be present widely in the genome. PMID- 6282894 TI - Cell behavior in Dictyostelium discoideum: preaggregation response to localized cyclic AMP pulses. AB - The motion of cells in the aggregation phase of Dictyostelium discoideum development is complex. To probe its mechanisms we applied precisely timed (+/- 1 s) and positioned (+/-2 micrometers) pulses of cyclic AMP to fields of cells of moderate density using a micropipette. We recorded cell behavior by time lapse microcinematography and extracted cell motion data from the film with our Galatea computer system. Analysis of these data reveals: (a) Chemotaxis lasts only about as long as the cyclic AMP signal; in particular, brief pulses (approximately 5 s) do not induce chemotaxis. (b) Chemotactic competence increases gradually from within an hour after the initiation of development (starvation) to full competence at approximately 15 h when aggregation begins under our conditions. (c) Cell motion reverses rapidly (within 20 s) when the external gradient is reversed. There is no refractory period for motion. We present a new description of the process of aggregation consistent with our result and other recent findings. (d) The behavioral response to cyclic AMP includes a phenomenon we call "cringing." In a prototypical cringe the cell speed drops within 3 s after a brief cyclic AMP stimulus, and the cell stops and rounds and then resumes motion after 25 s. (e) The development of the speed response in cringing as the cells age closely parallels the development of the cyclic AMP-induced light-scattering response of cells in suspension. (f) Cringing occurs in natural populations during weak oriented movement. The computerized analysis of cell behavior proves to be a powerful technique which can reveal significant phenomena that are not apparent to the eye even after repeated examination of the film. PMID- 6282896 TI - Inhibition of pinocytosis in rat embryo fibroblasts treated with monensin. AB - Rat embryo fibroblasts cultured in the presence of monensin exhibited an inhibited uptake of horseradish peroxidase. The inhibition was detected after 3 h, after which time the cells became increasingly vacuolated; the concentration of monensin required to inhibit pinocytosis (0.4 microM for half-maximum inhibition at 18 h) was similar to that found by others to inhibit secretion. Both the exchange of 5'-nucleotidase between the membranes of cytoplasmic organelles and the cell surface and the internalization of anti-5'-nucleotidase bound to the cell surface were inhibited by approximately 90% in monensin-treated cells. The effects of monensin were reversible: cells cultured first with monensin, and then in fresh medium, exhibited control levels of horseradish peroxidase uptake, exchange of 5'-nucleotidase, and internalization of anti-5' nucleotidase bound to the cell surface. After monensin treatment, the median density of both galactosyl transferase and 5'-nucleotidase increased from 1.128 to 1.148, and the median density of both N-acetyl-beta-glucosaminidase and horseradish peroxidase taken up by endocytosis decreased from 1.194 to 1.160. The results indicate that monensin is a reversible inhibitor of pinocytosis and, presumably, therefore, of membrane recycling. They suggest that the inhibition of membrane recycling occurs at a step other than the fusion of pinocytic vesicles with lysosomes and is perhaps a consequence of an effect of the ionophore on the Golgi complex. PMID- 6282895 TI - Hepatic Golgi fractions resolved into membrane and content subfractions. AB - Golgi fractions isolated from rat liver homogenates have been resolved into membrane and content subfractions by treatment with 100 mM Na2CO3 pH 11.3. This procedure permitted extensive extraction of content proteins and lipoproteins, presumably because it caused an alteration of Golgi membranes that minimized the reformation of closed vesicles. The type and degree of contamination of the fractions was assessed by electron microscopy and biochemical assays. The membrane subfraction retained 15% of content proteins and lipids, and these could not be removed by various washing procedures. The content subfraction was contaminated by both membrane fragments and vesicles and accounted for 5 to 10% of the membrane enzyme activities of the original Golgi fraction. The lipid compositions of the subfractions was determined, and the phospholipids of both membrane and content were found to be uniformly labeled with [33P]phosphate administered in vivo. PMID- 6282898 TI - Effects of retinoic acid on the binding and mitogenic activity of epidermal growth factor. AB - In this study the effects of retinoic acid on the binding and mitogenic activity of epidermal growth factor (EGF) in mouse fibroblast Balb/c 3T6 cells are further examined. Retinoic acid treatment of 3T6 cells results in a sixfold enhancement of 125I-labeled mouse EGF binding when assayed at 37 degrees C. In both retinoic acid-treated and control cells, cell-associated 125I-EGF is rapidly internalized, degraded, and secreted. Retinoic acid treatment does not seem to have a significant effect on the rate of internalization and degradation of EGF. At 0 degrees C, internalization of EGF is strongly inhibited in both retinoic acid treated and control cells. Under these conditions retinoic acid-treated cells still exhibit a tenfold higher level of EGF binding compared to control cells. When exposed to high concentrations of EGF both retinoic acid-treated and control cells "down-regulate" their EGF receptors. And although the growth rate of retinoic acid-treated cells is about half that of control cells, the rate at which EGF binding capacity is restored after down-regulation is about three times as fast as in control cells. No direct antagonism on EGF binding was observed between the tumor promoter 12-O-tetradecanoyl-phorbol-13-acetate (TPA) and retinoic acid. EGF is a potent mitogen for 3T6 cells in serum-free medium; retinoic acid inhibits the mitogenic activity of EGF even though it increases EGF binding. Retinoic acid also inhibits cell proliferation induced by sarcoma growth factor (SGF) and insulin. PMID- 6282899 TI - Changes of ornithine decarboxylase activity and polyamine content upon differentiation of mouse NB-15 neuroblastoma cells. AB - The possible functions of ornithine decarboxylase (ODC) and polyamines in the differentiation of mouse NB-15 neuroblastoma cells were investigated by examining the changes of these parameters in the differentiating and nondifferentiating NB 15 cells over a 5-day culture period. Differentiation of NB-15 cells was induced by the addition of dibutyryl cyclic AMP and 3-isobutyl-1-methylxanthine (1BMX) to the growth medium and was monitored by neurite outgrowth, increases of acetylcholinesterase (AChE), and RI cAMP-binding protein. Plating of NB-15 cells in fresh serum-containing growth medium was accompanied by rapid growth and a marked increase of ODC activity; this early increase of ODC activity was attenuated, both in duration and in magnitude, in the differentiating cells. The spermidine content of the differentiating neuroblastoma cells was significantly lower than that of the nondifferentiating cells. In the fully differentiated neuroblastoma cells, the ODC activity and spermidine content were lower than that of the undifferentiated cells by approximately 15-fold and five-fold, respectively. Based on these results it is proposed that changes of polyamine metabolism may be of significance in the differentiation of mouse neuroblastoma cells. PMID- 6282897 TI - Immunocytochemical localization of the receptor for asialoglycoprotein in rat liver cells. AB - We used high-resolution immunocytochemistry on ultrathin frozen sections labeled with colloidal gold to study the subcellular distribution of the asialoglycoprotein receptor in rat liver. The receptor was localized along the entire hepatocyte plasma membrane, including the bile capillary membrane, but was scarce intracellularly. Sinusoidal lining (Kupffer) cells and blood cells showed no immunoreactivity. In liver cells of rats injected with 1 to 100 micrograms of asialoorosomucoid (ASOR) 2-15 min before tissue fixation, endocytotic internalization of receptors at the blood front was conspicuous. At all times in this interval, receptor was present in approximately 100-nm vesicles and larger vacuoles adjacent to the sinusoidal plasma membrane. No other significant intracellular receptor was noted during the 15-min exposure to ASOR; in particular, lysosomes and Golgi complex were not labeled. Our observations, in combination with data from the literature which demonstrate that, under these conditions, the ligand is transferred further to the Golgi complex-lysosome region, suggest that the receptor and ligand are dissociated in the vicinity of the plasma membrane, after which the receptor rapidly returns to the cell surface. PMID- 6282900 TI - The effect of metabolic inhibition on ion contents and sodium exchange in human lymphocytes. AB - Lymphocytes depleted of ATP by incubation in iodoacetate (IAA) and nitrogen (N2) lost K and gained Na. Isotopic Na exchange showed a fast fraction and a slower exponential fraction, the latter conventionally assumed to reflect surface membrane properties. The gain of cell Na was not accounted for by a decrease in 22Na efflux in either the slow or the fast fraction. After 3-5 hours, Na efflux increased. These results led us to question the concept that normal cell ion levels are maintained by an ATPase pump and could not be explained by exchange diffusion, co-transport, countertransport, or other inherently dissipative mechanisms. The data are, on the other hand, consistent with the concept that cell ion contents are determined by their relative exclusion from cell water coupled with selective adsorption onto fixed macromolecular anionic sites within the cell. In this view, the IAA,N2-induced rise in cell Na is due to the occupancy of adsorption sites losing K, while the increased isotopic exchange is due to a decreased activation energy for ion-site interaction. PMID- 6282901 TI - Glucagon and choleragen stimulation of glycogenolysis in primary cultures of adult rat liver parenchymal cells: lack of involvement of the glucocorticoids. PMID- 6282902 TI - Effects of pyruvate on the growth of normal and transformed hamster embryo fibroblasts. AB - The growth of NIL and NILpy hamster embryo fibroblasts was determined in the presence and absence of pyruvate as a component of the growth medium. It was demonstrated that NIL cells respond to the presence of pyruvate by decreasing cell doubling time, glucose utilization, glutamine utilization, and increasing lactate production with the effects being more pronounced at low inoculum densities. Polyoma-virus transformed NIL cells (NILpy) demonstrate none of the above effects upon pyruvate addition regardless of initial cell inoculum density. PMID- 6282903 TI - Stimulation of thymidine kinase activity by chloramphenicol in Naegleria. AB - In the present study, the possible association of thymidine kinase (TK) with mitochondria in Naegleria was investigated by treating growing and differentiating cells with chloramphenicol (CAP), an inhibitor of mitochondrial protein synthesis. In some systems, CAP causes an overproduction of mitochondrial proteins coded for in the nucleus. The present results show that in growing Naegleria, CAP stimulates a dramatic increase in TK activity while growth and division is gradually inhibited. CAP does not stabilize the enzyme in vivo or in vitro. The stimulation is cycloheximide (CHI)-sensitive and specific since nucleoside phosphotransferase activity does not increase. In cells stimulated to differentiate, CAP does not prevent differentiation or the expected decrease in TK activity. Using polyacrylamide gel electrophoresis, a comparison of TK in mitochondrial and postmitochondrial fractions of CAP-treated and untreated cells was made. Results suggest some processing of the enzyme, resulting in a slight change in electrophoretic mobility. No mitochondrial TK was found. The stimulation of a cytoplasmic enzyme by CAP suggests a form of mitochondrial control of nuclear transcription for other than mitochondrial proteins. DNA synthesis in CAP-treated cells was not stimulated, suggesting (since TK and DNA synthesis are usually tightly coupled) an uncoupling of these two events, most likely, at the beginning of the S phase. PMID- 6282904 TI - Rous sarcoma virus transformed cells are resistant to cyclic AMP. AB - A nontransformed line of Chinese hamster ovary (CHO) cells (Pollard and Stanners, 1979) has been transformed by the Schmidt-Ruppin subgroup D strain of Rous sarcoma virus (SR-RSV). SR-RSV transformed CHO cells are shown to differ from spontaneously transformed cells in that the virally transformed cells are more resistant to growth inhibition or changes in cell shape by 8-Br-cyclic AMP or cholera toxin. SR-RSV transformed rat (NRK) cells also have a reduced sensitivity to growth inhibition by 8-Br-cyclic AMP. Cyclic AMP-dependent protein kinase was examined in SR-RSV transformed CHO cells, but no differences in enzyme level, activation by cyclic AMP, chromatographic behavior, or its ability to phosphorylate endogenous proteins in whole cells could be detected. It is concluded that transformation of CHO and NRK cells by SR-RSV alters the cells in a manner different from spontaneous transformation, and that this alteration does not affect cAMP-dependent protein kinase activity. PMID- 6282906 TI - Transport of 6-deoxy-D-glucose and D-xylose by untransformed and SV40-transformed 3T3 cells. AB - Transport rates of the nonphosphorylated D-glucose analogs 6-deoxy-D-glucose and D-xylose were measured in quiescent and serum-stimulated cultures of mouse 3T3 cells, in SV40-transformed 3T3 cells (SV101), and in a density revertant cell line derived from SV101 (FI-SV101). Initial rates of both entry and exit of 6 deoxy-D-glucose and D-xylose were more than threefold higher in serum-stimulated 3T3 and in SV101 cells than they were in quiescent 3T3 cells, but transport rates were not higher in the transformed cells (SV101) than they were in serum stimulated 3T3. Confluent cultures of FI-SV101 showed lower rates of transport than serum-stimulated FI-SV101, but not as low as quiescent 3T3 cells enter the quiescent G0 state, but emphasize that SV40-transformed 3T3 cells do not show higher activity of the D-glucose carrier than do actively growing 3T3 cells. Thus, enhanced glucose transport appears not to be a specific consequence of transformation, but a reflection of the active growth state of the cell. PMID- 6282905 TI - Myeloperoxidase-mediated oxidation of methionine. AB - The myeloperoxidase-mediated oxidation of methionine was studied using a purified canine myeloperoxidase preparation. The system required the simultaneous presence of myeloperoxidase, H2O2, and a halide anion. When 0.1 mM H2O2 was used, the system has a Ph optimum of approximately pH 5-5.5. Bromide and iodide were much more effective than chloride in the myeloperoxidase-mediated oxidation of methionine. Horseradish peroxidase was unable to oxidize methionine whether chloride or iodide was used. In contrast, lactoperoxidase oxidized methionine in the presence of iodide but not chloride. Based on studies of 1) the effect of various inhibitors and singlet oxygen quenchers, as well as 2) the effect of D2O on the oxidation of methionine, by the myeloperoxidase system, OCI-, or methylene blue, it was shown that the oxidation of methionine by the myeloperoxidase system was not mediated by OCI- or 1O2. The mechanism of the myeloperoxidase-mediated oxidation of methionine remains unclear. However, it may be one mechanism by which the myeloperoxidase system damage microorganisms. PMID- 6282907 TI - Altered cell cycle progression and aberrant mitosis in adenovirus-infected rodent cells. AB - Actively growing mouse or rat embryo cells suffered structural chromosome damage, mitotic anomalies, and polyploidy after infection by human adenovirus type 5. Chromosome damage required expression of one or more early viral genes and showed regular periodicity in its frequency. The growth cycle time of some of the infected cells was reduced by about 5 hours due to a decrease in G1, and the interval between successive waves of chromosome damage corresponded to this reduced cycle time. After infection there was a decrease in cells with G1 DNA content and an increase in cells with G2 diploid, aneuploid, and polyploid DNA contents. We suggest these effects are due to the expression in semipermissive cells fo early viral gene(s), whose function in productive infection in vivo is to alter cell cycle controls in order to maximize the number of cells able to replicate viral DNA and the time such cells spend in DNA replication. PMID- 6282908 TI - Characterization of a postlavage, in situ pulmonary macrophage population. AB - A postlavage in situ subpopulation of pulmonary macrophages (PM), biochemically distinct from the lavaged population, has recently been isolated from rats. After exhaustive bronchopulmonary lavage to extract the free lung cells, the lungs were excised, homogenized, and filtered, and the resultant cell suspension was allowed to form a monolayer on plastic Petri dishes. Electron microscopic morphometry failed to indicate any morphologic differences in the two populations. The postlavage in situ PM were more active metabolically during phagocytosis of zymosan particles or stimulation by phorbol myristate acetate (PMA) than the corresponding lavage population, as evidenced by greater superoxide generation. Macrophages prepared by either method became more avidly phagocytic when incubated with cell-free medium isolated in the preparation of the situ population. Peroxidase, an enzyme absent from the granules of PM separated by lavage techniques, was found in a granule-rich fraction of the in situ macrophage. Catalase activity was found in similar amounts in both supernatants and granule-rich fractions of both populations. The results support the concept of subpopulations of PM and suggest that these subpopulations are distinguished by their biochemical properties and their functional abilities. PMID- 6282909 TI - Increases in cyclic AMP potentiate competence formation in BALB/c-3T3 cells. AB - The ability of platelet-derived growth factor and fibroblast growth factor to stimulate the initiation of DNA synthesis in quiescent BALB/c-3T3 cells was enhanced by cholera toxin. However, the addition of cholera toxin to unsupplemented medium was not mitogenic, nor did cholera toxin increase the mitogenic potential of medium supplemented with platelet-poor plasma. The enhancement of serum-induced DNA synthesis by cholera toxin was due to a specific effect on competence formation and not plasma-controlled progression. Cholera toxin increased the rate of competence formation during a transient exposure of quiescent cells to platelet-derived growth factor; this rate was further increased by the addition of isobutylmethylxanthine, a cyclic nucleotide phosphodiesterase inhibitor. Intracellular cyclic AMP concentrations in quiescent BALB/c-3T3 cells were increased 2- to 3-fold after the addition of cholera toxin., The addition of cholera toxin plus 30 microM isobutylmethylxanthine caused an even greater (7- to 8-fold) increase in the cellular levels of cyclic AMP. That these increases in cyclic AMP concentrations mediated at least part of the increased sensitivity of quiescent cells to competence factors was substantiated by the observation that 0.01 to 1 mM monobutrylcyclic AMP or 8 bromocyclic AMP also caused a concentration-dependent potentiation of competence formation in quiescent cells during a transient exposure to platelet-derived growth factor. PMID- 6282910 TI - Benzo(a)pyrene-binding proteins of hamster embryo cell nuclei: comparison of nuclear isolation procedures. AB - Hamster embryo cells metabolize benzo(a)pyrene to derivatives that covalently modify the nuclear macromolecules including proteins. Not all proteins are modified to the same extent nor by the same metabolites. In particular, a protein of apparent molecular weight 32,000 is highly modified by derivatives of trans 9,10-dihydro-9,10-dihydroxy B(a)P. This protein is shown here to be preferentially lost from nuclei during purification by centrifugation through high molarity sucrose solutions followed by osmotic shock. It does not appear to be a cytoplasmic contaminant, but shares many properties of an abundant protein from Xenopus laevis oocytes, nucleoplasmin. PMID- 6282912 TI - A very precise high-performance liquid chromatographic procedure for the determination of cefmenoxime, a new cephalosporin antibiotic, in plasma. AB - A simple and very precise high-performance liquid chromatographic procedure has been developed for the determination of cefmenoxime, a new broad spectrum cephalosporin antibiotic, in plasma. The workup procedure involves ultrafiltration of samples which have been treated with sodium dodecyl sulfate to displace the drug from its binding sites on plasma proteins. The ultrafiltrates are then directly injected into a high-performance liquid chromatographic system utilizing a reversed-phase analytical column, and an ultraviolet spectrophotometric detector. The mean assay coefficient of variation over a concentration range of 0.5-200 micrograms/ml is slightly greater than 1% when either p-nitrobenzoic or p-anisic acid is used as the internal standard. Recoveries of drug are essentially quantitative at all levels investigated; hence the calibration curves are rectilinear from the limit of quantification (about 0.05 microgram/ml) to at least 200 micrograms/ml. PMID- 6282911 TI - Inhibition of neoplastic cell growth by quiescent cells is mediated by serum concentration and cAMP phosphodiesterase inhibitors. PMID- 6282913 TI - Improved high-performance liquid chromatographic assay for cimetidine using ranitidine as an internal standard. PMID- 6282914 TI - Rapid assay of cyclic AMP phosphodiesterase and 5'-nucleotidase by means of chromatography on cellulose-nitrate membrane strips. AB - A simple chromatographic procedure with the use of modified cellulose-nitrate membrane strips, 80 x 40 mm, has been worked out for the rapid isotopic assay of cyclic AMP (cAMP) phosphodiesterase (EC 3.1.4.17) and 5'-AMP nucleotidase (EC 3.1.3.5) in crude extracts of various tissues from animals and plants. The assay is based on enzymatic conversion of the product to adenine, a relatively inert compound which, in contrast to cAMP and 5'-AMP, is strongly adsorbed by the cellulose-nitrate membrane. Due to this property rapid separation of adenine from the unconverted substrate (cAMP or 5'-AMP) is possible. Commercial 5' nucleotidase and easily obtainable crude extract of adenosine nucleosidase from barley leaves are used as coupling enzymes for the phosphodiesterase assay. The assay of phosphodiesterase in 0.5-2 microliter of blood (10(-5) to 4.10(-5) units) has been demonstrated on several examples. PMID- 6282915 TI - Immunoelectroosmophoresis for the human papovavirus BK. AB - The reaction between BK virus and its antibodies may well be visualized with immunoelectroosmophoresis (IEOP). Serum antibody levels may be compared or graded by reacting them against a continuous antigen front. In seroepidemiologic surveys IEOP compares favourably with hemagglutination inhibition in terms of sensitivity, practicability and reproducibility. Technical conditions are described which are suitable for antigen detection, quantification and identification. PMID- 6282916 TI - A solid-phase radioimmunoassay for quantitative measurement of class-specific antibodies against tick-borne encephalitis virus. AB - Antigen-coated polystyrene spheres are used for a solid-phase RIA for IgM and IgG antibodies against tick-borne encephalitis virus (TBE). The use of highly purified anti-mu and anti-gamma antibodies permitted the construction of standard curves from which quantitative values for TBE-specific IgG and IgM could be obtained. An antibody-blocking test identifies non-specific results. PMID- 6282917 TI - Complement-fixation test for rotavirus detection: comparison and analysis of different methods to reduce anti-complementary activity of some specimens. AB - The complement-fixation test may be used to detect rotaviral antigens directly in clinical specimens. However, a certain number of specimens tested for human rotaviruses by the complement-fixation test show an anti-complementary activity. By comparing eight techniques we analysed this anti-complementary activity and identified the best method for its reduction. Pretreatment of clarified supernatant of stool suspensions by some methods resulted in a reduction of anti complementary activity, without reducing the sensitivity of the method. Clarified supernatants of 8/36 (22.2%) specimens were anti-complementary; this anti complementary activity was best removed by absorption with fetal calf serum or calf albumin. Such treatment offers practical means of increasing the specificity of complement-fixation test. Some observations suggest that the anti complementary activity of stool suspensions may be frequently due to presence of one or more chelating agents that may be in faecal specimens. PMID- 6282918 TI - Differentiation of foot-and-mouth disease virus strains using a competition enzyme-linked immunosorbent assay. AB - Foot-and-mouth disease virus isolates were compared using solid-phase competition and indirect microenzyme-linked immunosorbent assays. Results were compared to those obtained from complement fixation tests. Similar relationships between the isolates were obtained using the indirect enzyme immunoassay and complement fixation tests. The competition assay was more discriminatory and the results did not always correlate with the other two assays. PMID- 6282919 TI - Molecular epidemiology and carcinogen-DNA adduct detection: new approaches to studies of human cancer causation. PMID- 6282920 TI - Effects of dibutyryl adenosine 3',5'-monophosphate, luteinizing hormone-releasing hormone, and aromatase inhibitor on simultaneous outputs of progesterone, 17 beta estradiol, and human chorionic gonadotropin by term placental explants. AB - To contrast the effects of dibutyryl cAMP (dbcAMP) with those of LRH and to evaluate the effects of low density lipoprotein (LDL), dehydroepiandrosterone sulfate (DHEAS), and aromatase inhibitor (4-hydroxy-androst-4-ene-3,17-dione) on the output of hCG, 17 beta-estradiol, and progesterone, term human placental explants were maintained in culture for 6 days with daily changes of medium. The daily outputs of progesterone and hCG were observed to decrease while that of 17 beta-estradiol remained constant during the course of the incubation. The addition of 67 micrograms/ml LDL cholesterol had no effect on the basal output of 17 beta-estradiol, progesterone, or hCG. The addition of 4 micrograms/ml DHEAS increased 17 beta-estradiol output 20-fold, but did not affect the outputs of hCG or progesterone. The addition of 1.6, 3.2, or 6.4 micrograms/ml LRH had no effect on the output of progesterone or 17 beta-estradiol. LRH increased hCG output in Dulbecco's Modified Eagle's Medium with penicillin, streptomycin, insulin, and glucose alone, but not in the presence of added LDL or DHEAS, while dbcAMP (1, 2, and 4 mM) increased the output of hCG in all three media and decreased 17 beta estradiol output in medium supplemented with DHEAS. Aromatase inhibitor decreased both 17 beta-estradiol and hCG outputs in a dose-dependent fashion, but it was without effect on the output of progesterone. Basal progesterone, basal hCG, and dbcAMP-stimulated hCG outputs were unaffected by the addition of LDL or DHEAS. Both LDL and DHEAS inhibited the stimulatory effect of LRH on the output of hCG. Aromatase inhibitor decreased the output of both hCG and 17 beta-estradiol, but the effect on hCG was direct and not due to the decrease in 17 beta-estradiol. PMID- 6282921 TI - Multiple androgenic abnormalities, including elevated free testosterone, in hyperprolactinemic women. AB - To investigate the basis of the hirsutism and elevated plasma dehydroepiandrosterone (DHA) and/or DHA sulfate (DHAS) in hyperprolactinemic women, we measured androgen binding parameters and an extensive profile of plasma androgens in normal (NL) and hyperprolactinemic women (HYPRL). ACTH tests and dexamethasone (dex) suppression tests were performed in subgroups. Free testosterone levels were higher in HYPRL (13.1 +/- 23.3 vs. 7.18 +/- 0.72 pg/ml; P less than 0.025), although total testosterone was comparable. This disparity was related to plasma testosterone-estradiol-binding globulin (TEBG) levels being one third lower in HYPRL (mean +/- SE, 27.4 +/- 4.0 nM) than in NL (41.2 +/- 3.7 nM; P less than 0.0125). Less striking elevations of plasma DHAS, androstenedione, and 11-deoxycortisol were found in HYPRL. Plasma total dihydrotestosterone [17 beta-hydroxy-5 alpha-androstan-3-one (tDHT)] was nearly 30% lower in HYPRL (11.2 +/- 2.6 ng/dl) than in NL (15.6 +/- 1.3 ng/dl; P less than 0.025), whereas free DHT was normal. Ratios of tDHT to precursors were lower in HYPRL (P less than 0.005). After ACTH stimulation, hyperresponsiveness of 17 hydroxyprogesterone and androstenedione were observed. Apparent adrenal enzyme efficiencies, judged from post-ACTH product to precursor ratios, were normal in HYPRL with one exception: the ratio of tDHT to total testosterone at 4 h was lower (P less than 0.05). Dex suppression normalized androgens and obliterated the abnormal tDHT to precursor ratios. These findings suggest an ACTH dependency of the abnormalities. In summary, we find that about 40% of HYPRL have an androgenic abnormality, and the most characteristic abnormality is an elevated free testosterone level (abnormal in 43%). Depressed TEBG and high DHAS levels were found with lesser frequency (19-21%). The plasma tDHT concentration was low, both in absolute terms and relative to its precursors. Dex suppressibility of the hyperandrogenemia was also observed. We postulate that PRL may exert multiple effects on steroid secretion and metabolism. Possibilities include the inhibition of the TEBG level. PMID- 6282922 TI - Trophoblastic cells of the hydatidiform mole contain a beta 1-subtype adrenergic receptor. AB - Both beta 1- and beta 2-adrenergic receptors have been previously described in normal human placental homogenates; the cells upon whose surface membranes these receptors reside have not been identified. In order to show that a beta 1 adrenergic receptor is present on trophoblastic cells, the cells which mediate maternal-fetal transport and produce placental hormones, beta-adrenergic receptors were demonstrated in membrane fractions of human hydatidiform mole. Microscopic sections of the mole samples used demonstrated edematous villi lined by trophoblastic cells with minimal nontrophoblastic (stromal or vascular) contamination compared with placenta. (--)-[3H]Dihydroalprenolol [(--)-[3H]DHA] binding to molar membranes was reversible and saturable to a single class of sites (Kd = 0.97 +/- 0.12 nM; n = 7; maximum binding capacity, 72.9 +/- 6.4 fmol/mg protein). (--)-[3H]DHA binding was associated with catecholamine stimulated adenylate cyclase activity. Agonist competition for the molar beta adrenergic receptor showed the order of potency to be (--)isoproterenol much greater than norepinephrine = epinephrine, characteristic of a beta 1-adrenergic receptor subtype. Competition for (--)-[3H]DHA binding to trophoblastic membranes by the beta-adrenergic receptor subtype-specific agents metoprolol (beta 1 selective) and zinterol (beta 2 selective) was also characteristic of a homogeneous subtype of beta 1-adrenergic receptors. Because beta 1-adrenergic receptors alone were seen on trophoblast cells, the beta 2-adrenergic receptor in placenta must reside on nontrophoblastic elements (stromal or vascular endothelium). No differences in beta-adrenergic receptor binding were seen related with ploidy (2 or 3 N), the presence or absence of a fetus, or the progression of the mole to choriocarcinoma. Two choriocarcinoma cell lines, BeWo and JEG-3, however, showed no specific (--)-[3H]DHA binding. Human trophoblast contains beta 1-adrenergic receptors coupled to catecholamine-sensitive adenylate cyclase, supporting a role for catecholamines in the regulation of placental metabolism. PMID- 6282923 TI - Peripherally administered beta-endorphin increases cerebrospinal fluid endorphin immunoreactivity. PMID- 6282924 TI - Corticotropin-releasing factor-like activity in human placental extracts. PMID- 6282925 TI - Paradoxical responsiveness of adrenocorticotropin and cortisol to thyrotropin releasing hormone (TRH) in pregnant women. Evidence for intermediate lobe activity? PMID- 6282926 TI - Use of intravenous immune globulin in patients receiving bone marrow transplants. AB - Patients undergoing bone marrow transplantation for acute leukemia or aplastic are a great risk for the development of interstitial pneumonia associated with the cytomegalovirus (CMV). Passive immunization with a CMV immune plasma has been found to be effective for the prevention of interstitial pneumonia after marrow transplantation. Because of the limited availability of high-titered CMV plasma, we studied the kinetics of CMV antibody in bone marrow transplant patients receiving immune globulin intravenous, 5%, in 10% maltose (IGIV). Several lots of IGIV had CMV radioimmunoassay (RIA) antibody titers (1:30,000) comparable to the CMV antibody titers in the CMV immune plasma. A single infusion of 20 cc/kg of IGIV produced a mean peak CMV RIA antibody titer of 1:9,500. This titer fell to 1:1,000 after seven days. These antibody titers were approximately two-to three fold lower than the CMV RIA antibody titers achieved after a 10 cc/kg dose of the CMV immune plasma. Administration of IGIV at doses lower than 20 cc/kg produced correspondingly lesser increases in the CMV antibody titers of recipients. At all doses, IGIV was well tolerated and caused no significant biochemical abnormalities. Some patients experienced mild elevation of their serum glucose levels and asymptomatic glycosuria. We are presently evaluating the efficacy of IGIV (20 cc/kg given once every week) for the modification of CMV infection and prevention of interstitial pneumonia after bone marrow transplantation in a randomized, controlled study. The effects of IGIV on the incidence and outcome of bacterial, fungal, and other viral infections are also being analyzed. PMID- 6282927 TI - The effect of short-term and chronic immunosuppression on Theiler's virus demyelination. AB - Theiler's virus (TV)-infected mice were treated with antithymocyte serum (ATS), cyclophosphamide or pepstatin (a protease inhibitor) to determine the effect on demyelination. When ATS and cyclophosphamide were begun at the time of infection there was significantly less demyelination at 2.5-3.5 weeks than in pepstatin or non-treated infected controls. When immunosuppression was continued for 5 weeks, or when it was not started until 5 weeks post-infection, no significant decrease in demyelination was seen compared to controls. The findings indicate that timing of immunosuppression is critical in determining the extent of TV demyelination. Such demyelination may occur by different mechanisms that are active at different times. The "bystander effect' may be important in early demyelination, but late demyelination may be due to other causes, such as oligodendrocyte lytic infection. PMID- 6282928 TI - An ultrastructural and immunocytochemical study of astrocytic differentiation in vitro: changes in the composition and distribution of the cellular cytoskeleton. AB - Astroglia in cultures of dissociated neonatal rat optic nerves were studied by light microscopy, immunocytochemistry and electron microscopy to determine whether intermediate filaments play a role in defining the multipolar morphology of the mature astrocyte. Immature, polygonal astroblasts contained few glial filaments, in spite of exhibiting positive staining with antiserum against glial fibrillary acidic (GFA) protein. Microtubules were the most prominent cytoskeletal component at early stages of cytodifferentiation, but these were progressively reduced in number at later intervals and were gradually replaced by intermediate filaments. These observations suggest that microtubules are involved in the initial establishment of cytoplasmic asymmetry and process development. Subsequently, glial filaments may play a role in maintaining and stabilizing the overall geometry of the mature astrocyte. PMID- 6282929 TI - Experimental allergic encephalomyelitis in Lewis rats bearing avian sarcoma virus induced brain tumors. AB - Lewis rats bearing avian sarcoma virus (ASV)-induced brain tumors were injected with guinea pig spinal cord emulsion and complete Freund's Adjuvant to determine if they remained susceptible to induction of experimental allergic encephalomyelitis (EAE). The incidence of EAE among rats with small, and moderate sized gliomas was similar to non-tumor-bearing controls (P less than 0.5; P less than 0.9) while 18 of 24 (75%) animals with large gliomas developed EAE as compared to 31 of 33 (93%) controls (P less than 0.05). The histologic features and geographical distribution of "ordinary' EAE were seen in controls and were maintained in tumor-bearing rats. The presence of an intracranial tumor did not significantly alter the ability of Lewis rats to develop EAE. PMID- 6282930 TI - Central nervous system infection and immune response in mice inoculated into the lip with herpes simplex virus type 1. AB - Virus may be recovered from various areas of the central nervous system (CNS) of mice for as long as 11 days after inoculation of herpes simplex virus type 1 (HSV 1) into the lip. The probability of isolation from any particular region of the CNS seems to be a function of the distance of that area from the root-entry zone of the trigeminal nerve. It is also mouse strain-dependent, with much more extensive evidence of brain infection being found in BALB/c and C3H rather than C57BL/6 mice, in which it is limited to the pons. The virus could not be isolated from the CNS of BALB/c mice after 10 days, though HSV-1 is readily recovered from the trigeminal ganglia at least through day 38. Significant concentrations of HSV 1-specific immunoglobulin (Ig) were demonstrated consistently in cerebrospinal fluid (CSF) from day 12 after exposure to virus. The persistence of relatively high concentrations of IgM in the CSF indicates that much of this antibody may be synthesized locally in the brain. PMID- 6282931 TI - Inhibition of vascular permeability changes in rats by captopril. AB - Systemic treatment of rats with captopril (50 mg/kg body wt per os), a specific competitive inhibitor of angiotensin l-converting enzyme, significantly inhibits vascular permeability changes induced by the intradermal injection of the vasoactive mediators histamine, bradykinin, serotonin, and compound 48/80. This effect of captopril is both dose- and time-dependent with approximately 60% inhibition of edema formation observed 7 h after captopril treatment (100 mg/kg body wt per os). The inhibitory effect of captopril on edema formation is temporally unrelated to the inhibition of serum angiotensin l-converting enzyme activity or serum prostaglandin E2 levels and is not inhibited by systemic treatment of rats with indomethacin. The data suggest that captopril may have potent antiinflammatory activity through as yet undefined mechanisms. PMID- 6282933 TI - Primary cortisol resistance in man. A glucocorticoid receptor-mediated disease. AB - We have studied a man suspected of having primary cortisol resistance on the basis of high 24-h mean plasma cortisol levels (27.4 micrograms/dl) and no stigmata of Cushing's syndrome. His son had slightly elevated 24-h mean plasma cortisol levels (9.9 micrograms/dl; normal 7.52 micrograms/dl). Both had high plasma protein unbound cortisol and increased urinary free cortisol. Plasma ACTH concentration was high, and both were resistant to adrenal suppression by dexamethasone. The father appeared to have mineralocorticoid excess resulting in hypertension, hypokalemia, and metabolic alkalosis. This was found to be due to markedly elevated plasma levels of deoxycorticosterone and corticosterone. The son, who was normotensive, had mildly increased plasma corticosterone and normal deoxycorticosterone levels. To study the apparent end-organ resistance to cortisol, we examined the glucocorticoid receptor in the white cells and fibroblasts of these patients. In both tissues, using both whole cell and cytosol assays, the glucocorticoid receptor was found to have reduced affinity for dexamethasone. In the cytoxol assays, a reduced receptor number was found as well. We conclude that cortisol resistance is a rare familial syndrome owing to an abnormal glucocorticoid receptor with a decreased affinity for cortisol. PMID- 6282932 TI - Thrombin-induced exposure and prostacyclin inhibition of the receptor for factor VIII/von Willebrand factor on human platelets. AB - The receptor for Factor VIII/von Willebrand factor (F. VIIIVWF) is readily available on circulating platelets. We have found that the stimulation of platelets with traces of thrombin at concentrations that are generated physiologically (0.008 U-0.05 U/ml) induced concentration-dependent binding of 125I-labeled F. VIIIVWF in a steady-state system. The binding induced by thrombin was specific because it was inhibited by a 100-fold molar excess of unlabeled F. VIIIVWF factor, by rabbit monospecific antibody against Factor VIII, and was not inhibited by an excess of fibrinogen or fibronectin. Binding induced by thrombin required metabolically active platelets, in contrast to a system with ristocetin that also prompted binding to glutaraldehyde-treated platelets. The thrombin effects on binding of 125I-F. VIIIVWF was not observed when platelets were washed with EDTA-containing buffers; EDTA and EGTA both inhibited thrombin-induced binding. Platelet membrane glycoproteins were required because enzymatic stripping od them from the platelet surface with chymotrypsin reduced binding 2.5 5.0-fold. Prostacyclin, in the concentration range of 1 to 50 nM, had two distinct effects on the receptor for F. VIIIWVF: (a) it prevented exposure of this receptor when added 10 min before thrombin, and (b) it reversed the binding of 125I-F. VIIIVWF to the platelet receptor when added 30 min after thrombin and the ligand, ie., when binding was at equilibrium. The dual effect of prostacyclin on the receptor for F. VIIIVWF was reproduced by dibutyryl cyclic AMP. PMID- 6282935 TI - Human platelets contain gelsolin. A regulator of actin filament length. AB - Morphologic and biochemical studies suggest that actin in human platelets polymerizes in response to various stimuli and that shortening of actin filaments can be regulated by calcium. We report that human platelets contain gelsolin, a protein of Mr 91,000 that binds reversibly to actin in the presence of calcium. Platelet gelsolin exhibits immunologic crossreactivity with rabbit macrophage gelsolin and shortens actin filaments as demonstrated by viscosity measurements and gel point determinations. Gelsolin is active in micromolar calcium concentrations and its effects upon actin filaments are reversible. Gelsolin may be a dynamic regulator of actin filament length in the human platelet. PMID- 6282934 TI - Human skin collagenase in recessive dystrophic epidermolysis bullosa. Purification of a mutant enzyme from fibroblast cultures. AB - Recessive dystrophic epidermolysis bullosa, a genodermatosis characterized by dermolytic blister formation in response to minor trauma, is characterized by an incresaed collagenase synthesis by skin fibroblasts in culture. Since preliminary studies of partially purified recessive dystrophic epidermolysis bullosa collagenase suggested that the protein itself was aberrant, efforts were made to purify this enzyme to homogeneity, so that detailed biochemical and immunologic comparisons could be made with normal human skin fibroblast collagenase. Recessive dystrophic epidermolysis bullosa skin fibroblasts obtained from a patient documented to have increased synthesis of the enzyme were grown in large scale tissue culture and both serum-free and serum-containing medium collected as a source of collagenase. The recessive dystrophic epidermolysis bullosa collagenase was purified to electrophoretic homogeneity using a combination of salt precipitation, ion-exchange, and gel-filtration chromatography. In contrast to the normal enzyme, the recessive dystrophic epidermolysis bullosa collagenase bound to carboxymethyl-cellulose at Ca(2+) concentrations at least 10 times higher than those used with the normal enzyme. Additionally, this enzyme was significantly more labile to chromatographic manipulations, particularly when serum-free medium was used. However, rapid purification from serum-containing medium yielded a preparation enzymatically equivalent to normal human skin collagenase. Like the normal enzyme, the recessive dystrophic epidermolysis bullosa collagenase was secreted as a set of two closely related zymogens of approximately 60,000 and approximately 55,000 daltons that could be activated by trypsin to form enzymically active species of approximately 50,000 and approximately 45,000 daltons, respectively. Amino acid analysis suggested slight variations between the normal and recessive dystrophic epidermolysis bullosa collagenases. Cyanogen bromide digests demonstrated peptides unique to the enzyme from each source. The recessive dystrophic epidermolysis bullosa proenzyme was significantly more thermolabile at 60 degrees C than the normal, a finding that correlated with an approximate fourfold decrease in the affinity of the mutant enzyme for Ca(2+), a known activator and stabilizer of human skin collagenase. Aside from the altered affinity for this metal cofactor, kinetic analysis of the structurally altered recessive dystrophic epidermolysis bullosa collagenase revealed that its reaction rates and substrate specificity for human collagen types I-V were identical to those for the normal enzyme. Likewise, enzymes from both sources displayed identical energies of activation and deuterium isotope effects. Antisera were raised to the normal and putatively mutant procollagenases respectively, and, although they displayed a reaction of identity in double diffusion analysis, immunologic differences were present in enzyme inhibition and quantitative precipitation studies. These studies indicate that recessive dystrophic epidermolysis bullosa is characterized by the increased synthesis of an enzymically normal, but structurally aberrant, collagenase. PMID- 6282936 TI - Metabolic acidosis suppresses 25-hydroxyvitamin in D3-1alpha-hydroxylase in the rat kidney. Distinct site and mechanism of action. AB - Effect of metabolic acidosis on two distinct 25-hydroxyvitamin D(3)-1alpha hydroxylase (1alpha-hydroxylase) systems was studied in the kidneys of vitamin D deficient rats; one is localized in the proximal convoluted tubule (PCT), is activated in vitamin D deficiency, and is regulated primarily by parathyroid hormone (PTH) via cyclic AMP; the other is localized in the proximal straight tubule (PST), is latent in vitamin D deficiency, and is selectively stimulated by calcitonin via a cyclic AMP-independent mechanism. The 1alpha-hydroxylase activities were measured in the PCT and PST microdissected from the kidney of vitamin D-deficient rats with or without metabolic acidosis of varying duration. The 1alpha-hydroxylase activity decreased in the PCT from 0.74+/-0.07 fmol/mm per h to 0.24+/-0.02 at day 3 of metabolic acidosis without a further decline at day 7. Neither metabolic acidosis of 16 h duration nor reduction of the incubation medium pH from 7.4 to 7.0 affected the enzyme activity in the PCT. To examine the underlying mechanism for the suppression of 1alpha-hydroxylase activity, PTH, cyclic AMP, or calcitonin was given to rats with metabolic acidosis of 3 d duration. Although PTH failed to augment the suppressed 1alpha-hydroxylase activity in the PCT, cyclic AMP restored it to the level of control rats. The 1alpha-hydroxylase activity in the PST remained undetectable in control rats and in acidotic rats with or without PTH or cyclic AMP treatments. However, calcitonin stimulated the 1alpha-hydroxylase activity in the PST equally from undetectable to 0.75+/-0.09 fmol/mm per h in control and to 0.78+/-0.10 in acidotic rats. The data suggests that metabolic acidosis suppresses 1alpha hydroxylase only in the PCT by inhibiting PTH-dependent adenylate cyclase, and that cellular events beyond cyclic AMP in the PCT and the events responsive to calcitonin in the PST are unaffected. The results show the definite advantage of using defined single nephron segments to study the hormonal and ionic control of the 1alpha-hydroxylase system in the kidney. PMID- 6282938 TI - Binding of a calcium antagonist, [3H]nitrendipine, to high affinity sites in bovine aortic smooth muscle and canine cardiac membranes. AB - [(3)H]Nitrendipine, a potent calcium channel antagonist [3-ethyl-5-methyl-1-1,4 dihydro-2,6 - dimethyl - 4 - (3 - nitrophenyl) - 3,5 - pyridine carboxylate], was used to label high affinity binding sites on membranes prepared from bovine aortic smooth muscle. The binding of [(3)H]nitrendipine is rapid (t(1/2) < 5 min) and reversible at 37 degrees C. The binding sites have a high affinity for [(3)H]nitrendipine with an equilibrium dissociation constant of 2.1 nM. The density of sites is 40-60 fmol/mg of membrane protein. Analogues of nitrendipine compete for the binding sites with affinities consistent with their known biological effects as calcium antagonists. Nisoldipine, [isobutyl methyl 1,4 dihydro-2,6-dimethyl-4-(2-nitrophenyl)-3,5-pyridine carboxylate], a calcium antagonist more potent than nifedipine [2,6-dimethyl-3,5-dicarbomethoxy-4-(2 nitrophenyl)-1,4-dihydropyridine] in relaxing vascular smooth muscle, has an affinity three-fold higher than that of nifedipine in competing for the binding sites. A biologically inactive derivative of nifedipine does not compete for [(3)H]nitrendipine binding. Verapamil (alpha-isopropyl-alpha[(N-methyl - N homoveratryl) -alpha-aminopropyl]-3,4-dimethyoxyphenyl acetonitrile), a structurally different calcium antagonist, only partially (25%) inhibits binding at high concentrations (1 muM). Prazosin, an alpha adrenergic antagonist does not compete for [(3)H]nitrendipine binding sites. The binding of [(3)H]nitrendipine is not affected by 1.5 mM calcium. Canine cardiac membranes also have high affinity [(3)H]nitrendipine binding sites, (K(D) = 6 nM) but bovine erythrocytes do not. The relative affinities of nisoldipine and nifedipine for the cardiac membrane binding sites reflect the relative activities of these compounds as calcium channel antagonists. These results suggest that the [(3)H]nitrendipine binding sites are the sites through which dihydropyridines act as calcium channel antagonists. PMID- 6282937 TI - Hypertriglyceridemic very low density lipoproteins induce triglyceride synthesis and accumulation in mouse peritoneal macrophages. AB - Triglyceride-rich lipoproteins may be responsible for the lipid accumulation in macrophages that can occur in hypertriglyceridemia. Chylomicrons and very low density lipoproteins (VLDL, total and with flotation constant [S(f)] 100-400) from fasting hypertriglyceridemic subjects induced a massive accumulation of oil red O-positive inclusions in unstimulated peritoneal macrophages. Cell viability was not affected. The predominant lipid that accumulated in cells exposed to hypertriglyceridemic VLDL was triglyceride. Hypertriglyceridemic VLDL stimulated the incorporation of [(14)C]oleate into cellular triglyceride up to ninefold in 16 h, but not into cholesteryl esters. Mass increase in cellular triglyceride was 38-fold. The stimulation of cellular triglyceride formation was dependent on time, temperature, and concentration of hypertriglyceridemic VLDL. By contrast, VLDL, low density, and high density lipoproteins from fasting normolipemic subjects had no significant effect on oleate incorporation into neutral lipids or on visible lipid accumulation.(125)I-Hypertriglyceridemic VLDL (S(f) 100-400) were degraded by macrophages in a dose-dependent manner, with 50 and 100% saturation observed at 3 and 24 mug protein/ml (2.5 and 20 nM), respectively. Hypertriglyceridemic VLDL inhibited the internalization and degradation of (125)I hypertriglyceridemic VLDL (4 nM) by 50% at 3 nM. Cholesteryl ester-rich VLDL from cholesterol-fed rabbits gave 50% inhibition at 5 nM. Low density lipoproteins (LDL) inhibited by 10% at 5 nM and 40% at 47 nM. Acetyl LDL at 130 nM had no effect. We conclude that the massive triglyceride accumulation produced in macrophages by hypertriglyceridemic VLDL is a direct consequence of uptake via specific receptors that also recognize cholesteryl ester-rich VLDL and LDL but are distinct from the acetyl LDL receptor. Uptake of these triglyceride-rich lipoproteins by monocyte-macrophages in vivo may play a significant role in the pathophysiology of atherosclerosis. PMID- 6282939 TI - Ultrasonic demonstration of small pancreatic islet cell tumors. PMID- 6282940 TI - Ovarian tumours of Wolffian or allied nature: their place in ovarian oncology. AB - Two unusual ovarian tumours thought to be of Wolffian identity, one of them malignant, are described. They showed packed combinations of adenopapillary, tubular, trabecular and diffuse patterns, a sharp and generalised periodic acid Schiff (PAS)-positive basement membrane and areas of elastic network. A review of published Wolffian tumours at various sites suggests that the prototypes of the two tumours occur chiefly in the cervix and broad ligament. The significance of Wolffian tumours and their differentiation from arrhenoblastoma and serous tumours is discussed. PMID- 6282941 TI - Determination of specific IGA antibodies to varicella zoster virus by immunoperoxidase assay. AB - An indirect peroxidase technique was developed for determination of IgA antibodies to varicella zoster virus (VZV). The antigen consisted of acetone fixed trypsinised VZV-infected cells. Rabbit antihuman IgA peroxidase conjugate was used to detect human IgA antibodies bound to viral antigen. In parallel IgG antibodies to VZV were determined by an immunoperoxidase antibody to membrane antigen (IPAMA) technique. Varicella zoster virus IgA antibodies were detected in all five varicella and seven zoster patients. No VZV IgA antibodies (less than 2) were detected in 45 healthy control sera. Neither were they found in paired sera of five patients with herpes simplex infection, five patients with human cytomegalovirus infection and two patients with Epstein-Barr virus infection. Application of immunoperoxidase IgA technique in serodiagnosis of primary and reactivated VZV infections is discussed. PMID- 6282942 TI - Experimental infection of broiler chickens with an avian reovirus. PMID- 6282943 TI - Assessment of phosphonoformate-treatment of pigeon herpesvirus infection in pigeons and budgerigars and Aujeszky's disease in rabbits. PMID- 6282945 TI - The pathogenesis of nephritis in chickens induced by infectious bronchitis virus. PMID- 6282944 TI - Immunosuppression in toxoplasmosis: studies in lambs and sheep infected with louping-ill virus. PMID- 6282946 TI - The successful treatment of pityriasis versicolor by systemic ketoconazole. PMID- 6282947 TI - Effect of increasing the bulk content of the diet on the rat parotid gland and saliva. AB - The present study, along with others, shows that when the requirement of a diet for mastication is increased, enlargement of the parotid gland occurs. This study demonstrated for the first time that the magnitude of the gland enlargement is directly related to the amount of nonnutritive bulk incorporated into the diet, and that the concentration of protein is significantly increased in the parotid saliva of rats fed such a diet. The findings show that, although the concentration of protein is increased in the parotid saliva, the composition of proteins is unchanged, leading to the conclusion that all secretory proteins are increased in parallel. A comparison of the gland enlargement observed in response to the bulk diet with that observed in response to chronic isoproterenol treatment reveals many similarities. However, the fact that the protein composition of the parotid saliva is not altered when rats are fed the bulk diet is in marked contrast to the striking changes observed following chronic isoproterenol treatment. The reason for the dissimilar response is not known. However, an investigation into the differences that exist in the gland response to these two means of inducing gland enlargement may lead to a better understanding of the factors that regulate the size of the rat parotid gland and the complement of proteins in its secretion. PMID- 6282948 TI - In vitro hydrolysis of monofluorophosphate by dental plaque microorganisms. AB - Enzymic hydrolysis of sodium monofluorophosphate by suspensions of dental microorganisms has been demonstrated at pH 5.1, pH 7.0, and pH 8.4, using a fluoride-selective electrode. The extracellular medium from viable Streptococcus mutans K1R cells contained low MFPase and paranitrophenyl phosphatase activity. It is hypothesized that the enzymes responsible for MFP hydrolysis by S. mutans K1R are intracellular, and that cell disruption is necessary for hydrolysis to be manifested; this question requires further study. In vitro MFPase activity was of a magnitude consistent with the hypothesis that it may significantly raise the fluoride ion concentration of plaque within the several minutes MFP would be in the mouth during toothbrushing. PMID- 6282949 TI - Isocyanate-induced pulmonary diseases: a current perspective. PMID- 6282950 TI - Current concepts about the pathogenesis of silicosis and asbestosis. AB - Silicosis and asbestosis are two forms of fibrotic lung disease resulting from the inhalation of inert materials indigestible by pulmonary alveolar macrophages. Results of studies of the host response to these particulates have not always been consistent. It is clear, however, that after phagocytosis, both cause alveolar macrophage damage, with resultant release of macrophage products, including fibrogenic factors and chemotactic factors for neutrophils. The latter cells also release lysosomal enzymes and free radicals when exposed to silica and asbestos. The net effect of these observations suggests that the combination of tissue damage and fibroblast stimulation results in the pulmonary fibrosis characterizing these diseases. Patients with silicosis and asbestosis have normal or decreased cell-mediated and increased humoral immunity with a high incidence of circulating immune complexes and autoantibodies. Whether these abnormalities are related to the pathogenesis of pulmonary fibrosis or are epiphenomena remains to be determined. PMID- 6282951 TI - Calculates vs. analyzed composition of four modified fat diets. Formulated to study effects in human subjects of kind and amount of dietary fat. AB - Four diets, varying in kind and amount of fat, were calculated to meet the needs of adult men. Commonly available U.S.A. food were used. Fourteen days of menus, each with five caloric levels varying from 2,400 to 4,000 kcal, were calculated. Chemical analyses of composites were carried out to determine proximate composition, food energy, selected vitamins and minerals, sugars, fatty acids, and sterols. The diets were used in a controlled study of the effects of fat level and P:S on blood pressure, blood lipids, platelet aggregation, and other variables. In general, good agreement was found between calculated and analyzed nutrient contents. However, analyzed values for fiber, iron, cholesterol, and vitamin C showed larger than desirable differences from calculated values. PMID- 6282952 TI - Phytate:zinc molar ratio, mineral, and fiber content of three hospital diets. AB - Phytate:zinc molar ratio, minerals, and fiber contents of regular, ovo-lacto vegetarian, and soy meat-substitute hospital diets were determined. The phytate:zinc molar ratio, minerals, and fiber contents of the soy meat-substitute diet were significantly higher than those of ovo-lacto vegetarian and regular hospital diets is within acceptable limits established by human and animal studies conducted by the USDA's Vitamin and Mineral Nutrition Laboratory. PMID- 6282953 TI - Pseudo-Cushing's syndrome: the role of alcohol. AB - The mechanisms responsible for the genesis of pseudo-Cushing's syndrome are poorly understood. We studied the effect of acute ethanol administration on the hypothalamic-pituitary-adrenal axis in five chronic alcoholic patients and three normal volunteers. Ethanol was administered alone and together with beta 1-24 ACTH given intravenously. Baseline plasma ACTH and cortisol concentrations were significantly higher in the patients than in normal controls (p less than 0.05). Acute ethanol administration produced no significant change in the mean plasma ACTH concentration, failed to augment the effect of exogenous ACTH on the adrenal gland, and produced similar changes in the serum cortisol and aldosterone concentration in both the normal controls and the alcoholic patients. We speculate that pseudo-Cushing's syndrome may represent a state of stress-induced hypercortisolemia secondary to multiple episodes of subacute withdrawal from ethanol. PMID- 6282955 TI - A new method for preparation of cleansed surface of dental hard tissues. (Water glass embedding method). PMID- 6282954 TI - Serological study of Japanese encephalitis outbreak in Deoria District of Uttar Pradesh. PMID- 6282956 TI - Alkaline treated surfaces of human enamel using the water glass embedding method. (The external wall of rod and the pseudo rod sheath). PMID- 6282957 TI - Some scanning electron microscopic observations of dead tracts and peritubular cementum by water glass embedding method. PMID- 6282958 TI - Scanning electron microscopic observation of the peritubular dentin and the interglobular dentin using the water glass embedding method. PMID- 6282960 TI - Presence of polioviruses and other enteral viruses in sewage: a survey in the Czech Socialistic Republic 1969-1976. AB - Monitoring polioviruses in sewage is a permanent task within the poliomyelitis vaccination control programme in Czechoslovakia. The results of sewage examination in certain localities of the CSR in the years 1969 to 1976 are reported. During the study years, nearly 3600 samples of sewage were examined. The yield was about 9 percent of samples positive for polioviruses and 22 per cent of samples positive for other viruses. The virus positivity rate was roughly the same in samples from municipal sewers and samples from children's facilities. The majority of polioviruses were detected within 3 to 4 months after mass vaccination campaigns, which in Czechoslovakia are carried out in spring. In some years types 2 and 3 polioviruses tended to occur in later months after vaccination as well. Antigenic characterization of the polioviruses isolated in Prague showed that polioviruses possessing an rct+ marker and antigenically related but not identical with the original vaccine strains were sometimes present late during the intervals between vaccination campaigns. It may be assumed that vaccine strains which have been in circulation for sufficiently long time are subject to a gradual change or that vaccine-like strains have been imported. PMID- 6282959 TI - Rabbit nasopharyngeal colonization by Bordetella pertussis: the effects of immunization on clearance and on serum and nasal antibody levels. AB - Two Bordetella pertussis antigen preparations, outer membrane protein (OMP) and filamentous haemagglutinin (FHA), and a standard vaccine were used to immunize rabbits, and the effects on nasopharyngeal colonization by the organism were determined. Antibodies were measured in serum and in nasal washes by ELISA before and after challenge of the rabbits with 10(6) bacteria of strain M2. Recoveries of B. pertussis in nasal washes were used to assess colonization, which in controls persisted for at least 65 days. Some rabbits of all the immunized groups showed enhanced clearance, but there was no correlation between the elimination of B. pertussis and serum antibodies to OMP, FHA, lipopolysaccharide, lymphocytosis-promoting factor or agglutinogen 3. In contrast, nasal IgA antibody to FHA showed significant inverse correlation with bacterial persistence. Such antibody was induced by the OMP preparation as well as by FHA, but to different extents depending on the immunization schedule and adjuvant used. PMID- 6282961 TI - Study of persistence of maternal antibodies to varicella-zoster virus by indirect haemagglutination, with result control by radioimmunoassay. AB - The persistence of maternal antibody against varicella-zoster virus was studied by the method of indirect haemagglutination (IH) and a control of the results was performed by radioimmunoassay (RIA). The findings were analysed with reference to the constant half-life of passively acquired IgG and the titre range and frequency of antibodies in 220 persons of fertile age representative of the normal population. Statistical analysis of expected and the experimentally obtained data was performed. Among the normal population, the expectancy is a gradual decrease in maternal IH antibody titres to negativity during the first 10 months of life and 100% sero-negativity by the 11th and 12th month. The experimental data showed a decrease in IH seropositivity from the initial 100% in 1-month infants to 27% in 6-month and 7% in 12-month infants. Nine out of twelve of the experimentally found positive titres in infants during the second half of their first year of life deviated from the normal population sample. The results obtained by both of the methods used were in good agreement. PMID- 6282962 TI - Excretion of attenuated polioviruses in children vaccinated with live oral poliovirus vaccine. AB - The excretion of attenuated polioviruses was studied in a group of nursery children vaccinated with 105TCD50 of each type of virus. The primovaccinated children were found to excrete type 1 poliovirus for 8 weeks, type 2 for 11 weeks after the vaccination with the type 1 + 2 bivaccine. Poliovirus type 1 as eliminated by 78% and type 2 by 98% of the vaccinees. The separately administered type 3 was detectable for 6 weeks and was isolated from 100% of the vaccinees. The highest per cent of children with type 1 excretion positivity was recorded at week 5, with type 2 positivity at week 1 and with type 3 positivity at week 2. The poliovirus excretion peaked early after the vaccination, the titres of the poliovirus type 2 were the highest. The children revaccinated next year with the type 1 + 2 bivaccine eliminated the respective types of virus 1 - 2 weeks; type 3 poliovirus was detectable for 6 weeks after revaccination and was excreted by the highest per cent of vaccines. The contact infections caused by the attenuated polioviruses developed in 9 from 22 children vaccinated previously. The excretion of polioviruses did not last longer than 1 week. The contact infections were most frequently caused by the poliovirus type 2. The examined children, particularly those vaccinated previously, turned out to excrete also other enteroviruses identified as Coxsakieviruses B 4 and B 5 and Echovirus 21. In the primovaccinated these viruses were isolated only from those with the negative excretion of polioviruses. PMID- 6282963 TI - Specific in vitro antibody responses by human blood lymphocytes: apparent nonresponsiveness of PBL is due to a lack of recirculating memory B cells. PMID- 6282964 TI - Mechanisms of human cell-mediated cytotoxicity. I. Modulation of natural killer cell activity by cyclic nucleotides. PMID- 6282965 TI - Mechanisms of human cell-mediated cytotoxicity. II. Correction of the selective defect in natural killing in the Chediak-Higashi syndrome with inducers of intracellular cyclic GMP. PMID- 6282966 TI - Immune complex induced generation of oxygen metabolites by human neutrophils. AB - Human neutrophils have been studied for their ability to respond with production of O(2) and H2O2 by human neutrophils contain a 1:2 weight ratio of antigen and antibody (molar ratio of 1.5:1). The metabolic stimulation of leukocytes is a linear function of the total amount of complex used. Complexes containing F(ab')2 are ineffective in stimulating leukocytes to produce O(2) and H2O2. The complexes that maximally stimulate production of O(2) by neutrophils differ from those complexes that are 1) most effective in complement fixation, 2) maximally taken up by neutrophils, and 3) most effective in the induction of enzyme release. These findings suggest that immune complex-induced damage in tissues may reflect the effects of a heterogeneous population of immune complexes. PMID- 6282967 TI - Cell-mediated immune response in experimental visceral leishmaniasis. II. Oxygen dependent killing of intracellular Leishmania donovani amastigotes. PMID- 6282969 TI - Partial removal of sialic acid enhances phagocytosis and the generation of superoxide and chemiluminescence by polymorphonuclear leukocytes. PMID- 6282968 TI - Protection of neonatal mice against herpes simplex virus infection: probable in vivo antibody-dependent cellular cytotoxicity. AB - Infant mice are extremely susceptible to fatal Herpes simplex virus (HSV) infection. They are unable to produce antibody to HSV, and their leukocytes cannot mediate antibody-dependent cellular cytotoxicity (ADCC) to HSV-infected cells. In order to avoid H-2-dependent effector mechanisms and instead analyze possible in vivo ADCC, a murine model employing adoptive transfer of antibody and human leukocytes was developed. Administration of either human immune globulin or leukocytes i.p. from HSV immune or nonimmune humans could not protect infant C57BL/6 mice from fatal HSV infection. In contrast, a combination of a subneutralizing dilution of globulin and leukocytes from nonimmune or immune human donors, given one day before inoculation, was highly protective against lethal HSV infection. The cells involved included lymphocytes or monocyte macrophages. At least 5 X 10(6) viable leukocytes (or 1 X 10(6) monocyte macrophages) and immune serum globulin concentrations as low as 10(-8) were protective. Infected cell monolayer adsorption and DEAE column fractionation demonstrated that the protection by globulin was due to specific antiviral IgG antibody. Protection was n ot seen in animals receiving virus before immune transfer. Protection did not involve synergistic viral neutralization by antibody and cells, as shown by in vitro experiments. Animals receiving globulin and cells, unlike normal infant mice, had circulating antiviral antibody and peritoneal leukocytes able to mediate ADCC to HSV-infected cells. This is the first in vivo evidence for the role of human ADCC. This model also allows for the in vivo evaluation of the ability of cells from immunocompromised humans to curb viral infection. PMID- 6282970 TI - Immunologic abnormalities of the autoimmune mouse, Palmerston North. PMID- 6282971 TI - A role for oxygen-dependent mechanisms in killing of Leishmania donovani tissue forms by activated macrophages. AB - Leishmania donovani, the causative agent of visceral leishmaniasis, infects macrophages (M phi ) of susceptible vertebrates. Immunologically activated M phi are leishmanicidal, but the mechanisms involved in the killing process are not well defined. We sought to investigate the role of reactive oxygen intermediates in the killing of L. donovani. Both the free-swimming promastigote and the intracellular amastigote forms were found to be susceptible to killing in vitro by hydrogen peroxide and other oxygen intermediates. Upon phagocytosis by mouse peritoneal M phi, promastigotes elicited a significantly stronger respiratory burst compared with amastigotes as measured by release of superoxide anion. Although amastigotes do not elicit a strong burst of M phi oxidative metabolism during the initial phagocytic event, immunologically activated M phi that acquired leishmanicidal capacity could be triggered to release substantial amounts of H2O2. Hence, the development of leishmanicidal capacity was correlated temporally with enhanced H2O2 generation by the M phi. In contrast, M phi that lost their ability to release significant amounts of H2O2 after several days in culture were unable to eliminate their parasite burden. Catalase markedly inhibited the elimination of amastigotes by lymphokine-stimulated M phi. In toto, the results implicate reactive oxygen intermediates in killing of the tissue form of L. donovani by its host cell, the mononuclear phagocyte. PMID- 6282972 TI - Circulating immune complexes, antigens, and antibodies related to the murine mammary tumor virus in C3H mice. PMID- 6282973 TI - Bystander suppression of tumor growth: evidence that specific targets and bystanders are damaged by injury to a common microvasculature. PMID- 6282974 TI - CT hybridomas: tumor cells capable of lysing virally infected target cells. AB - By fusing primed murine lymphocytes with a syngeneic T cell lymphoma, we have been able to select for H-2-restricted, virus-specific cytotoxic T cell hybridomas (CTH). These T cell hybrids, which replicate in ordinary tissue culture medium or in ascites, are capable of lysing virally infected target cells, and their activity is facilitated by the presence of lectins in the assay medium. Unlike cells mediating lectin nonspecific lysis, these hybridomas are H-2 restricted and specific for single viral proteins. The ability to maintain these cells in culture for over 18 mo and to pass them in vivo without loss of activity or specificity indicates that they will provide sufficient material for the analysis of surface proteins and genetic information required for the recognition and lysis of virally infected cells by killer T cells. PMID- 6282975 TI - Specific antisera produced by direct immunization with slices of polyacrylamide gel containing small amounts of protein. AB - Rabbits were injected with slices of polyacrylamide gels containing entrapped insect proteins after separation by electrophoresis. Specific antibodies were produced independently of the nature of the gel (with or without sodium dodecyl sulphate) and of the staining technique (amido black or Coomassie Blue). The procedure appears to be a rapid and simple method for production of antibodies specific to proteins separated in minute quantities from a complex mixture. PMID- 6282976 TI - Congenital liposarcoma. PMID- 6282978 TI - Adrenal function in women with idiopathic acne. AB - The adrenal secretion of androgens were examined in 9 women (ages 19-39 yr) with postadolescent idiopathic acne and compared to age and sex-matched normal controls. Plasma dehydroepiandrosterone (DHA), dehydroepiandrosterone sulfate (DHAS), androstenedione (delta 4-delta), cortisol, 17-hydroxyprogesterone, 11 deoxycortisol, and testosterone were measured by radioimmunoassay in the basal state and during a 48 hr ACTH infusion. The mean plasma and time-integrated plasma levels of the 3 adrenal androgens in patients with acne were 15-25% higher than normal controls, but the groups were not significantly different (p greater than .05). The plasma testosterone values, on the other hand, were similar in both groups. In addition, cortisol, 11-deoxycortisol and 17-hydroxyprogesterone basal plasma values and responses to ACTH in patients with acne were similar to the normal control values. These findings suggest that adrenal androgen secretion is at most mildly elevated in patients with idiopathic acne and is unlikely to be the sole cause of acne since many patients without acne have similar hormone levels. Increased sensitivity of the sebaceous gland to androgens or increased local metabolism of androgen hormones in the skin to potent androgen metabolites may offer alternative mechanisms for the pathogenesis of this disorder. PMID- 6282977 TI - Embryonal adenocarcinoma in sacrococcygeal teratoma. PMID- 6282979 TI - Partial purification and characterization of angiotensin-converting enzyme in mouse and human skin. AB - The chemical properties of skin angiotensin-converting enzyme were characterized in mouse and human. In newborn mice, dermis contained almost all activity (1.3 mU/mg), of which 30% was in the 22,000 kappa g supernatant and 70% in the pellet which could be solubilized by Triton X-100. The activity increased sharply during the first 6 weeks after the birth. By gel filtration, the enzyme in the supernatant was Mr 330,000, but the solubilized enzyme was Mr 430,000 in the presence of Triton X-100. By electrophoresis, the 2 enzyme fractions demonstrated a charge difference. The enzymes showed the same pH optima of around 8.1 and 7.7, and Km values of 2.6 and 0.11 mm for Hip-His-Leu and angiotensin I, respectively. Sensitivity to known inhibitors and heat stability of the enzymes in 2 fractions were similar. In the human, 56% of the activity in skin homogenates (1.5 mU/mg) appeared in this supernatant and 44% in the pellet. Both soluble and solubilized preparations showed enzyme activity with 2 different molecular weights, 330,000 and 430,000. The human enzymes had chemical properties similar to the mouse enzymes, but their affinity for Hip-His-Leu and angiotensin I were 1.6 and 2 times higher than those of the mouse enzymes. PMID- 6282980 TI - Superoxide production by eosinophils: activation by histamine. AB - Both guinea pig peritoneal exudate and human peripheral blood eosinophils produce large amounts of superoxide anion when stimulated by preopsonized zymosan or phorbol myristate acetate (PMA). Superoxide production is also activated by histamine but not the histamine metabolite, imidazole acetic acid. Supernatants from degranulated rat mast cells stimulate superoxide production. In studies of both human and guinea pig eosinophils, the H1-antagonist, chlorpheniramine (10-3 M and 10-4 M), preopsonized zymosan histamine) production of superoxide anion but the H2-antagonist, cimetidine, only modestly inhibited superoxide anion production (zymosan, PMA), These studies provide direct evidence for the influence of histamine on the oxidative metabolism of eosinophils. These results are consistent with the hypothesis that histamine interacts with eosinophils predominantly via an H1 receptor site. Furthermore, they suggest that eosinophils may participate in immediate hypersensitivity reactions by the release of superoxide anion in response to stimulation by histamine. PMID- 6282981 TI - Abnormalities in collagenase expression as in vitro markers for recessive dystrophic epidermolysis bullosa. AB - In order to correlate a characteristic clinical phenotype with biochemical abnormalities in recessive dystrophic epidermolysis bullosa, fibroblast cultures were established from 4 typical patients with the severe form of the disease. Collagenase, the enzyme implicated in the pathogenesis of blistering, was present in vitro in 2- to 4-fold greater concentrations than in control fibroblast cultures. Partially purified preparations of this enzyme displayed marked thermal lability and diminished affinity for Ca2+, a metal cofactor, suggesting the existence of a mutant enzyme. The data suggest that these 3 biochemical abnormalities, increased synthesis, decreased thermal stability and diminished affinity for Ca2+, should serve as reliable in vitro markers for genetic discrimination of recessive dystrophic epidermolysis bullosa. PMID- 6282982 TI - Collagenases and collagen degradation. AB - Degradation of interstitial collagens probably takes place through different enzymatic mechanisms than degradation of basement membrane and pericellular collagens. Interstitial collagens are resorbed under pathological and physiological conditions by collagenases which function extracellularly and cleave polypeptide chains in the collagen triple helix at specific loci resulting in solubilization from the fibril. Production of collagenase in humans is ascribable to fibroblast-like cells which can be stimulated to synthesize new enzyme for release outside of the cell. In several inflammatory conditions, such as rheumatoid synovitis, modulation of collagenase production is mediated by interactions with surrounding inflammatory cells. Monocyte-macrophages produce a stimulatory factor, which has homologies with interleukin 1, which not only increases collagenase synthesis but also PGE2 synthesis. The PGE2 in turn has profound effects on cellular function. Production of the mononuclear cell factor is modulated by several interactions including T lymphocytes and T lymphocyte products, collagen of the extracellular matrix and the Fc portion of immunoglobulins. It is probable, from analogies with other stimulants such as phorbol myristate acetate, that the increase in collagen synthesis is controlled at the level of transcription. Further regulation of collagenase action outside of the cell is probably accomplished by proteolytic activation of a latent collagenase zymogen and interactions with inhibitory proteins also produced by cells in the local environment of the resorptive process. PMID- 6282983 TI - Herpes simplex virus encephalitis: laboratory evaluations and their diagnostic significance. AB - Laboratory procedures were compared with brain biopsy findings in 113 biopsy proven patients with herpes simplex virus (HSV) encephalitis and 93 biopsy negative individuals. Examinations of brain tissue by histopathology, immunofluorescence, and electron microscopy demonstrated evidence of HSV infection in 56%, 70%, and 45% of proven cases and apparently false-positive results in 14%, 9% and 2% of those biopsy-negative. Serologic assessments revealed that HSV encephalitis occurred as both a primary (30%) and recurrent (70%) infection. Among patients with HSV encephalitis, 28% failed to seroconvert or seroboost within one month of the onset of disease. Titers of passive hemagglutinating and IgG immunofluorescent antibodies increased fourfold in the cerebrospinal fluid in 74% and 94%, respectively, of patients with proven disease. Similar percentages of patients had antibody ratios in serum and cerebrospinal fluid of less than 20 over the same interval. These data indicate the need for development of noninvasive diagnostic procedures. PMID- 6282984 TI - Shedding of infected ciliated epithelial cells in rhinovirus colds. AB - Specimens of nasal mucus were collected from 17 volunteers experimentally infected with rhinovirus and from 10 uninfected control subjects. Ciliated epithelial cells (CECs) were found in the nasal mucus of 16 of the infected volunteers but of none of the control subjects. The CECs were shown to contain rhinovirus antigen by indirect immunoperoxidase and fluorescent antibody staining. The pattern of CEC shedding was similar to the mean nasal symptom score pattern of the volunteers. However, the amount of CED shedding for an individual was not correlated with the severity of the symptoms. This finding suggests that the symptoms of a rhinovirus cold may be produced in part by processes that are independent of the severity of direct virus-induced damage to the epithelium. PMID- 6282985 TI - Epidemiology of diarrhea due to rotavirus on an Indian reservation: risk factors in the home environment. AB - Analysis of outpatient visits for diarrheal disease at the San Carlos Apache Reservation, Arizona, during 1977-1979 revealed a sharp increase in cases of infantile gastroenteritis of unknown etiology in October and November of each year. During the 1980 autumn diarrhea season, 19 patients and 12 control subjects were selected from among outpatients under two years of age and were interviewed and studied for bacterial and viral enteric pathogens. Eleven (58%) of 19 patients and two (17%) of 12 control subjects were positive for fecal rotavirus by enzyme-linked immunosorbent assay (P = 0.03). Ten (91%) of 11 rotavirus positive patients were under one year of age. The most significant risk factor for illness was the presence of a household contact under the age of two years (P - 0.004). Dog ownership was also associated with rotavirus infection (P = 0.05). PMID- 6282986 TI - Effect of acyclovir on genital herpes in guinea pigs. AB - The chemotherapeutic efficacy of acyclovir was evaluated in guniea pigs that were inoculated intravaginally with herpes simplex virus type 2 (HSV-2). Acyclovir was administered systemically (50 mg/kg of body weight per day ip) beginning three days after inoculation and continuing for 10-11 days. Treatment with acyclovir reduced the incidence of paralysis of the hind limbs, mortality, and the severity and duration of genital lesions but had little effect on the excretion of virus from the genital tract. Recovery of HSV-2 from neural tissues of infected animals during latent infection was less frequent in acyclovir-treated animals than in untreated ones. These data suggest that acyclovir may be a promising drug for altering the severity of clinical genital herpes in humans during acute infection and reducing the incidence of viral latency following primary infection. PMID- 6282987 TI - Excretion of cytomegalovirus in mothers: observations after delivery of congenitally infected and normal infants. AB - Shedding of cytomegalovirus (CMV) was studied in 142 women who gave birth to congenitally infected infants and in 81 seropositive control mothers (mothers of uninfected infants). Viral cultures from the throat, vagina, and urine were obtained at intervals between one month and nearly 12 years after delivery. In both groups the prevalence of excretion of CMV was greater in younger women and fell to low levels by age 30. Considering all of the sites of infection, 60% of the mothers of infected infants were shedding CMV within the first three months post partum compared with 18% of the control mothers; CMV shedding rates declined during the first 12 months post partum to 35% in the former group and to 3% in the latter. More than three years after delivery, seven (15%) of 45 mothers who transmitted CMV still had viruria. The excretion of CMV is common and persistent in mothers of children with congenital infections due to CMV. PMID- 6282988 TI - Centers for Disease Control. Enteroviral disease in the United States, 1970-1979. PMID- 6282989 TI - Diarrhea due to Norwalk virus in families. PMID- 6282990 TI - Rotavirus infections and the sompe syndrome. PMID- 6282991 TI - An epidemic of acute hemorrhagic conjunctivitis due to coxsackievirus A24. AB - An epidemic of acute hemorrhagic conjunctivitis in 416 (10.8%) of 3,830 staff and students of the Christian Medical College and Hospital in Vellore, India, during September-December 1979 was studied. Virus was isolated in cultures of HeLa cells from 171 of the 249 persons cultured. Most of the viral isolates were identified as an antigenic variant of coxsackievirus A24. Success in isolating the virus from conjunctival swabs was inversely proportional to the time elapsed between the onset of illness and the time the specimens were collected. In general, titers of antibody to coxsackievirus A24 were low or undetectable. Three persons had subconjunctival hemorrhage with no other symptoms, and virologic evidence of infection with coxsackievirus A24 was obtained in two of them. PMID- 6282992 TI - Rapid multiple-determinant enzyme immunoassay for the detection of human rotavirus. AB - Enzyme immunoassays (EIAs) can be used to detect rotavirus antigens accurately in human stool specimens. Conventional EIA systems require prolonged incubations and multiple washing steps, thus making them impractical for truly rapid diagnosis. However, more rapid solid-phase EIA systems can be devised which make use of simultaneous binding of enzyme-labeled and solid-phase antibodies at immunologically distinct antigenic sites. A multiple-determinant EIA system was devised that was capable of detecting small quantities of rotavirus antigen in less than 40 min. This assay system accurately detected rotavirus antigen in 45 stool specimens positive for rotavirus. No false-positive reactions were noted provided that appropriate control reactions were performed. The rapid double determinant EIA system provides an accurate, objective means for the rapid diagnosis of human viral infections. PMID- 6282993 TI - Survival of influenza viruses on environmental surfaces. AB - To investigate the transmission of influenza viruses via hands and environmental surfaces, the survival of laboratory-grown influenza A and influenza B viruses on various surfaces was studied. Both influenza A and B viruses survived for 24-48 hr on hard, nonporous surfaces such as stainless steel and plastic but survived for less than 8-12 hr on cloth, paper, and tissues. Measurable quantities of influenza A virus were transferred from stainless steel surfaces to hands for 24 hr and from tissues to hands for up to 15 min. Virus survived on hands for up to 5 min after transfer from the environmental surfaces. These observations suggest that the transmission of virus from donors who are shedding large amounts could occur for 2-8 hr via stainless steel surfaces and for a few minutes via paper tissues. Thus, under conditions of heavy environmental contamination, the transmission of influenza virus via fomites may be possible. PMID- 6282994 TI - The diagnostic significance of the indirect haemagglutination test in amaebiasis in children. PMID- 6282995 TI - [An immunohistochemical study in conjunction with the enzyme labelled antibody method to discern the relevancy between the localization of hCG, hPL and the functions of the normal and malignant trophoblast (author's transl)]. PMID- 6282996 TI - Effects of sex steroid hormones on rat anterior pituitary LH-RH receptor. AB - In order to elucidate the mechanism of estrogen action at the anterior pituitary level, the effects of estradiol on the number of binding sites of LH-RH receptor in the anterior pituitary as well as its effects on the anterior pituitary responsiveness to LH-RH were studied in castrated adult female rats. Daily sc injections of 20 micrograms of 17 beta-estradiol for 2 and 4 days increased both LH-RH receptor levels and the responsiveness of the pituitary to LH-RH. This result indicates that estrogen modulates the sensitivity of the anterior pituitary to LH-RH by changing its LH-RH receptor. PMID- 6282998 TI - [The prognostic evaluation of clinical course in sarcoidosis by initial serum angiotensin converting enzyme level (author's transl)]. PMID- 6282997 TI - [Maintenance chemotherapy for ovarian adenocarcinoma with carboquone: doses and side effects (author's transl)]. PMID- 6282999 TI - Paraganglioma of the liver seven years following the resection of a carotid body tumor - a case report. PMID- 6283000 TI - Corticosteroid production in abetalipoproteinemia: evidence for an impaired response ACTH. AB - The concept that an inherent absence of plasma LDLs may be associated with a reduced synthesis of steroid hormones has been evaluated in two patients with ABL. Basal production of adrenal corticosteroids, assessed by the concentrations of serum cortisol and the rates of excretion of urinary 17OHCS and 17KS and urine free cortisol, was normal in both patients with ABL. Prolonged stimulation (24 to 36 hr) with ACTH, however, disclosed an impairment in adrenal corticosteroid production in both patients with ABL (as compared to normolipidemic controls), which was manifest by lower serum cortisol levels, reduced rates of urinary excretion of 17OHCS and 17KS and a marked decrease in the excretion of urine free cortisol. These results provide in vivo evidence to support the view that plasma LDLs serve as an important source of cholesterol for adrenal corticosteroid synthesis during prolonged stimulation with ACTH but show that a total absence of LDL does not impair adrenal steroidogenesis in the basal state. PMID- 6283002 TI - Extranasopharyngeal angiofibromas. (Sex incidence and age distribution). PMID- 6283001 TI - Correlation of in vitro and in vivo effects of gold compounds on leukocyte function: possible mechanisms of action. AB - The anti-inflammatory effects of gold compounds include suppression of PMN lysosomal enzyme release. Since lysosomal products can provoke PMN aggregation, we assessed the effect of two gold compounds, auranofin and GST, on suppressing aggregation, degranulation, and metabolic functions of the cells. Aggregation of 1 x 10(7) cytochalasin B-treated PMNs in response to 2 x 10(-7)M FMLP, as assessed by light scattering, was inhibited in a dose-dependent fashion by both drugs. Concentrations of auranofin ranging from 5 to 20 microM caused 30.8% to 89% inhibition, whereas 200 microM GST reduced aggregation by only 32%. FCS or BSA added to suspensions of normal PMNs considerably reduced the gold compound inhibitory effect on PMN aggregation. Cell viability assessed by dye exclusion and lactate dehydrogenase release was unaffected by the drugs. The suppressive activities of the drugs could not be removed by washing the PMNs. Correspondingly, the drugs suppressed lysosomal enzyme release induced by FMLP of PMNs rendered secretory with cytochalasin B. Concentrations of 20 microM auranofin and 200 microM GST resulted, respectively, in a 61.5% and 19.3% reduction of release of lysozyme, 61.7% and 27.1% reduction of beta glucuronidase, 84.8% and 33.7%s reduction of myeloperoxidase, and 50.0% and 25.0% reduction of lactoferrin. Furthermore, auranofin inhibited 14C-1-glucose oxidation through the hexose monophosphate shunt in response to stimulation by either PMA or methylene blue. The in vivo studies suggested that auranofin could prevent neither neutropenia induced by zymosan-activated serum nor a corresponding rise in plasma lactoferrin levels. These findings suggest that the beneficial effect of gold compounds in rheumatoid arthritis are unlikely to be related to their ability to dampen PMN activation in vivo. PMID- 6283003 TI - Update on nutrition therapy for diabetes. PMID- 6283004 TI - Effects of prolactin-secreting tumour on copulatory behaviour in male rats. AB - The effect of the transplantable prolactin- and ACTH-secreting tumour 7315a on male copulatory behaviour was investigated. Castrated tumour-bearing rats implanted with testosterone-filled capsules exhibited significantly longer latencies to first mount or intromission and to ejaculation than castrated control animals. In contrast to the number of intromissions, the number of mount before ejaculation of the tumour-bearing rats was considerably increased. However, when castration was carried out in addition to adrenalectomy, the differences in copulatory behaviour between tumour-bearing rats and control rats were no longer present. During the last tests for copulatory behaviour the tumour bearing rats had serum prolactin concentrations or more than 4000 ng/Ml while control rats had less than 100 ng/ml. Plasma testosterone levels produced by silicone elastomer capsules were neither affected by the presence of the tumour nor by adrenalectomy. It was concluded that hyperprolactinaemia does not suppress the copulatory behaviour of adrenalectomized rats. PMID- 6283005 TI - Hepatic receptors for insulin and glucagon in relation to plasma hormones and metabolites in pregnant and unmated ewes. AB - Glucagon and insulin receptors were examined in relation to plasma concentrations hormones and metabolites in unmated and in 110- and 140-day pregnant ewes (four animals per group). The concentrations of insulin, growth hormone and non esterified fatty acids in the circulation, together with the maintenance of body weight, suggested that the animals were in energy surplus. When compared with the unmated group the binding insulin to isolated hepatocytes increased by 110 days of pregnancy, attaining statistical significance (P less than 0.02) after 140 days. Conversely, glucagon binding was reduced by 110 days of pregnancy, also attaining statistical significance (p less than 0.02) after 140 days. The changes in both insulin and glucagon binding were primarily due to changes in the number of receptors on each hepatocyte, although some fluctuation in receptor affinity were also found. These observations suggested that the number of hepatic insulin and glucagon receptors are altered during the metabolic demands of pregnancy in sheep, but unlike the changes reported during lactation in the ewe and restricted energy intake in the goat, they are not related either to energy deficit or to changes in the concentration on insulin, and probably of glucagon, in the circulation. PMID- 6283006 TI - Fate of human corticotrophin immediately after intravenous administration to the rat. AB - The distribution and degradation of tritium-labelled human 1-39 corticotrophin have been studied after intravenous administration to the rat. Within 5 min of injection of single major metabolite, 3-39 corticotrophin, appears in the circulation. This metabolite, however, has only 3.6% of the steroidogenic potency of 1-39 corticotrophin and the evidence suggests that it is formed in muscle and skin. By 5 min, extensive degradation had occurred in all the major tissues in the body (muscle, skin, liver, kidney, gut). PMID- 6283007 TI - Oestrogen induction of progestin receptors in the rat brain and pituitary gland: quantitative and kinetic aspects. PMID- 6283008 TI - Peptides derived from pro-opiocortin in the pituitary gland of the dogfish, Squalus acanthias. AB - Combined rostral, median and neurointermediate lobes from 650 pituitary glands of the dogfish Squalus acanthias were extracted in 0.1 M-HCl and subjected to filtration on Sephadex G-50. Fractions were monitored for ACTH, alpha-MSH, gamma MSH and endorphin by heterologous radioimmunoassay, corticotrophin-like intermediate lobe peptide (CLIP) and gamma-MSH by homologous radioimmunoassay, and methionine enkephalin after enzymatic digestion. The majority (about 99%) of the immunoreactivity detected was present as small peptides, with alpha-MSH, CLIP and gamma-MSH predominant. The single peaks of ACTH, alpha-MSH and CLIP contrasted with many distinct peaks of endorphin-like immunoreactivity. The gamma MSH radioimmunoassays monitored different peptides; the heterologous assay revealed a single major peak, while the homologous assay detected a number of distinct small peptides. The results suggested that there may be more than three distinct forms of MSH in the dogfish pituitary gland. Small amounts of much larger proteins were detected by a number of the radioimmunoassays, suggesting that peptides related to ACTH, gamma-MSH and lipotrophin are derived from common pro-opiocortin-type precursor molecules in this phylogenetically ancient cartilaginous fish. PMID- 6283009 TI - Human skin androgen metabolism and preliminary evidence for its control by two forms of 17 beta-hydroxysteroid oxidoreductase. AB - Human forehead skin incubated in vitro is known to metabolize testosterone to 17 oxosteroids faster than the reverse reaction, while axillary skin rapidly metabolizes androstenedione to 17 beta-hydroxysteroids, such as testosterone and 5 alpha-dihydrotestosterone. While this has been confirmed using a larger number of patients, some indication has been found that 17 beta-hydroxysteroid oxidoreductase activity declines with age in the axilla. The relative rates of 17 beta-oxidation and reduction (direction of operation of skin 17 beta hydroxysteroid oxidoreductase activity) were not altered by variety of incubation conditions. Large amounts of a membrane-bound 17 beta-hydroxysteroid oxidoreductase, showing preference for NAD as coenzyme and testosterone (rather than androstenedione) as steroid substrate, were found in forehead skin from one patient. On the other hand, the main axillary skin enzyme in skin from another patient was soluble and showed preference for NADP and androstenedione. It is postulated that 17 beta-oxidation and reduction in skin is controlled by the relative amount, the coenzyme preferences and the kinetic properties of these two enzymes. PMID- 6283010 TI - Terminal care. PMID- 6283011 TI - Differentiation of Dictyostelium discoideum cells in suspension culture. AB - Differentiation of Dictyostelium discoideum cells in suspension culture is reported, using a medium containing glucose, albumin, cyclic AMP, EDTA and streptomycin in a phosphate buffer. Production of UDPgalactose:polysaccharide transferase, an enzyme specifically present in prespore cells, and the formation of prespore-specific antigens in more than 60% of the cells, are demonstrated. Differentiation in this medium differs from that previously reported with other suspension systems in that (a) cells form only small, amorphous agglomerates, (b) there is an absolute requirement for cyclic AMP and (c) prior formation of loose cell mounds on a solid substratum is essential for subsequent differentiation in this medium. This last requirement indicates that the differentiation process, giving rise to the prespore-specific enzyme and antigen, can be resolved into two distinct stages, one requiring cell contact on a solid substratum and the other proceeding in small agglomerates incubated in the medium. This medium may be useful for elucidating the role of cyclic AMP and cell contact in slime mould development. PMID- 6283012 TI - Streamers: chemotactic mutants of Dictyostelium discoideum with altered cyclic GMP metabolism. AB - Mutants of Dictyostelium discoideum that developed huge aggregation streams in expanding clones were investigated using optical and biochemical techniques. Representatives of the six complementation groups previously identified (stmA stmF) were found to be similar to the parental wild-type strain XP55 in both the extent and timing of their ability to initiate and relay chemotactic signals and in the formation of cyclic AMP receptors and phosphodiesterases. The mutants differed from the wild-type in producing an abnormal chemotactic (movement) response visible using both dark-field optics with synchronously aggregating amoebae on solid substrata and light scattering techniques with oxygenated cell suspensions. Mutants of complementation group stmF showed chemotactic movement responses lasting up to 520 s, rather than 100 s as seen in the parental and other strains. Measurements of cyclic GMP formed intracellularly in response to chemotactic pulses of cyclic AMP in stmF mutants showed that abnormally high concentrations of this nucleotide were formed within 10 s and were not rapidly degraded. A causal correlation between defective cyclic GMP metabolism and the altered chemotactic response is suggested, and a model is proposed that accounts for the formation of huge aggregation streams in clones of these mutants.U PMID- 6283013 TI - Control of pyruvate kinase activity during glycolysis and gluconeogenesis in Propionibacterium shermanii. AB - The concentrations of glycolytic intermediates and ATP and the activities of certain glycolytic and gluconeogenic enzymes were determined in Propionibacterium shermanii cultures grown on a fully defined medium with glucose, glycerol or lactate as energy source. On all three energy sources, enzyme activities were similar and pyruvate kinase was considerably more active than the gluconeogenic enzyme pyruvate, orthophosphate dikinase, indicating the need for regulation of pyruvate kinase activity. The intracellular concentration of glucose 6-phosphate, a specific activator of pyruvate kinase in this organism, changed markedly according to both the nature and the concentration of the growth substrate: the concentration (7-10 mM) during growth with excess glucose or glycerol was higher than that (1-2 mM) during growth with lactate or at growth-limiting concentrations of glycerol or glucose. Other glycolytic intermediates, apart from pyruvate, were present at concentrations below 2 mM. Glucose 6-phosphate overcame inhibition of pyruvate kinase activity by ATP and inorganic phosphate. With 1 mM ATP and more than 10 mM inorganic phosphate, a change in glucose 6-phosphate concentration from 1-2 mM was sufficient to switch pyruvate kinase from a strongly inhibited to a fully active state. The results provide a plausible mechanism for the regulation of glycolysis and gluconeogenesis in P. shermanii. PMID- 6283014 TI - The effects of cyanide, azide, carbon monoxide and salicylhydroxamic acid on whole-cell respiration of Acanthamoeba castellanii. PMID- 6283015 TI - Generation of a membrane potential by one of two independent pathways for nitrite reduction by Escherichia coli. AB - A set of four isogenic Escherichia coli strains has been constructed in which all possible combinations of NADH- and formate-dependent nitrite reductases are active or inactive. Each pathway can be inactivated genetically without a corresponding loss in the other activity: the two pathways are therefore biochemically independent. The generation of a membrane potential during nitrite reduction by formate has been demonstrated using an ion-selective electrode specific for a lipophilic cation. The observed energy conservation results, at least in part, from the ability of formate dehydrogenase in E. coli to pump protons. PMID- 6283016 TI - 22Na+ uptake and catecholamine secretion by primary cultures of adrenal medulla cells. AB - The uptake of 22Na+ and secretion of catecholamines by primary cultures of adrenal medulla cells under the influence of a variety of agonists and antagonists were determined. Veratridine, batrachotoxin, scorpion venom, and nicotine caused a parallel increase in 22Na+ uptake and Ca2+-dependent catecholamine secretion. Ba2+, depolarizing concentrations of K+, and the Ca2+ ionophore Ionomycin stimulated secretion of catecholamines but did not increase the uptake of 22Na+. Tetrodotoxin inhibited both 22Na+ uptake and catecholamine secretion evoked by veratridine, batrachotoxin, and scorpion venom, but had no effect on 22Na+ uptake and catecholamine secretion caused by nicotine. On the other hand, histrionicotoxin, which blocks the acetylcholine receptor-linked ion conductance channel, blocked nicotine-stimulated 22Na+ uptake and catecholamine secretion, but only partially inhibited veratridine-stimulated catecholamine secretion and had no effect on veratridine-stimulated 22Na+ uptake. The combination of veratridine plus tetrodotoxin, which has been shown to prevent nicotine-stimulated secretion of catecholamines by adrenal medulla cells, also prevented nicotine-stimulated 22Na+ uptake by the primary cultures. These studies demonstrate the presence of tetrodotoxin-sensitive Na+ channels in adrenal medulla cells which are functionally linked to Ca2+-dependent catecholamine secretion. However, These channels are not utilized for Na+ entry upon activation of nicotinic receptors; in this case Na+ entry occurs through the receptor associated ion conductance channel. PMID- 6283018 TI - Pharmacological and electrophysiological characterization of lithium ion flux through the action potential sodium channel in neuroblastoma X glioma hybrid cells. AB - Interaction of Li+ with the voltage-dependent Na+ channel has been analyzed in neuroblastoma X glioma hybrid cells. The cells were able to generate action potentials in media containing Li+ instead of Na+. The uptake of Li+ into the hybrid cells was investigated for the pharmacological analysis of Li+ permeation through voltage-dependent Na+ channels. Veratridine and aconitine increased the uptake of Li+ to the same degree (EC50 30 microM). This increase was blocked by tetrodotoxin (IC50 20 nM). Veratridine and aconitine did not act synergistically; however, the veratridine-stimulated influx was further enhanced by the toxin of the scorpion Leiurus quinquestriatus (EC50 0.06 micrograms/ml). This stimulation was also blocked by tetrodotoxin. Thus, the voltage-dependent Na+ channel of the hybrid cells accepts both Li+ and Na+ in a similar manner. PMID- 6283017 TI - Metabolism of phospholipids in peripheral nerve from rats with chronic streptozotocin-induced diabetes: increased turnover of phosphatidylinositol-4,5 bisphosphate. AB - The effect of chronic streptozotocin-induced diabetes on phospholipid metabolism in rat sciatic nerve in vitro was investigated. In normal nerve incubated for 2 h in Krebs-Ringer-bicarbonate buffer containing [32P]orthophosphate, radioactivity was primarily incorporated into phosphatidylinositol-4,5-bisphosphate and phosphatidylcholine. Smaller amounts were present in phosphatidylinositol-4 phosphate, phosphatidylinositol, and phosphatidic acid. As compared to controls, phosphatidylinositol-4,5-bisphosphate in nerves from animals made diabetic 2, 10, and 20 weeks earlier accounted for 30-46% more of the isotope, expressed as a percentage, incorporated into all phospholipids. In contrast, the proportion of radioactivity in phosphatidylcholine decreased by 10-25%. When the results were expressed as the quantity of phosphorus incorporated into phospholipid, only phosphatidylinositol-4,5-bisphosphate displayed a change. The amount of isotope which entered this lipid increased 60% and 67% for 2- and 10-week diabetic animals, respectively. Increased phosphatidylinositol-4,5-bisphosphate labeling was observed when epineurial-free preparations were used or when the composition of the incubation medium was varied. Sciatic and caudal nerve conduction velocities were decreased after 10 and 20 weeks but were unchanged after 2 weeks. We conclude that an increase in the turnover of phosphatidylinositol-4,5 bisphosphate in sciatic nerve from streptozotocin-diabetic rats appears relatively early and persists throughout the course of the disease. This metabolic alteration may be related to a primary defect responsible for the accompanying deficient peripheral nerve function. PMID- 6283019 TI - Ontogeny of adenosine binding sites in rat forebrain and cerebellum. AB - The metabolically stable adenosine analogue N6-cyclohexyl [3H]adenosine ([3H]CHA) was used to label adenosine receptors in rat forebrain and cerebellum during development. [3H]CHA binding develops rather slowly, with adult binding levels obtained at 24 days in cerebellum and later in the forebrain. Ontogenic profiles in both areas are consistent with the onset of neuronal differentiation. High and low affinity sites appear to develop in parallel, since Scatchard analysis in forebrain tissue obtained from 5-day-old animals revealed both binding sites. PMID- 6283020 TI - Role of neuronal signal input in the down-regulation of central noradrenergic receptor function by antidepressant drugs. AB - Rats with unilateral lesions of the locus coeruleus were used to study the role of norepinephrine (NE) signal input in the down-regulation by antidepressants of the noradrenergic cyclic AMP-generating system in the cortex. Chronic administration of both desipramine (blockade of NE reuptake) and iprindole (no blockade of NE reuptake) reduced the cyclic AMP response to NE on the nonlesioned side, but had little or no effect on the lesioned side. The results indicate that NE signal input and thus the formation of the NE-receptor complex are prerequisites for inducing noradrenergic subsensitivity. PMID- 6283021 TI - Binding of [3H]gamma-aminobutyric acid and [3H]muscimol in purified rat brain synaptic plasma membranes and the effects of bicuculline. PMID- 6283023 TI - Ouabain-sensitive and ouabain-resistant net uptake of potassium into astrocytes and neurons in primary cultures. AB - Inhibition of net uptake of 42K by different concentrations of ouabain was studied in primary cultures of astrocytes and in primary cultures of neurons in order to investigate whether there is a pronounced difference between ouabain sensitivity in the two cell types and to determine the genuine magnitudes of the ouabain-sensitive and the ouabain-resistant potassium uptakes. In morphologically differentiated astrocytes, obtained after treatment with dibutyryl cyclic AMP (dBcAMP), the sensitivity to ouabain was slightly lower than in neurons, but astrocytes which had not been treated with dBcAMP showed sensitivity similar to the neurons (which likewise were not treated). In the presence of elevated potassium concentrations (12 and 24 mM) ouabain sensitivity was decreased, although only by a factor of 2-3. Accordingly, maximum inhibition of the uptake required under all conditions studied, at most, 1.0 mM ouabain. Like total uptake, this ouabain-sensitive uptake was several times less intense in neurons than in astrocytes, where it reached its maximum value at an external potassium concentration of 12 mM. Subtraction of the ouabain-sensitive uptake from the total uptake revealed a considerable ouabain-resistant uptake. This ouabain resistant uptake was studied in detail in the astrocytes, where it was found to increase with increasing potassium concentration over the whole concentration range 3-24 mM and to exceed substantially the maximum amount that can be accumulated by diffusion. PMID- 6283022 TI - Neuroblastoma X glioma hybrid cells synthesize enkephalin-like opioid peptides. AB - Partially purified extracts from neuroblastoma X glioma hybrid cells 108CC15 inhibit, like opioids, the prostaglandin E1-evoked formation of cyclic AMP in a dose-dependent manner in the same hybrid cells. The inhibition is prevented by the opioid antagonist naloxone. In addition, the same extract competes with [3H]naloxone and [3H]Leu-enkephalin for binding to opioid receptors of hybrid cell membranes and to a specific antiserum, respectively. The opioid activity in the extracts is destroyed by carboxypeptidase A and leucine aminopeptidase, but not by trypsin. Further purification of the extracts by HPLC, TLC, or high voltage paper electrophoresis reveals in each case two active fractions which behave like Met- and Leu-enkephalin. The Met-enkephalin-like, but not the Leu enkephalin-like, fraction is inactivated by treatment with BrCN. Dimethylaminonaphtylsulfonyl (dansyl) derivatives of Met- and Leu-enkephalin correspond to [3H]dansyl derivatives of Met-like substances from hybrid cells. Three to four times as much Met-enkephalin-like as Leu-enkephalin-like material is present in the extract. The overall concentration of opioid peptides in the hybrid cells varies between 0.03 and 1.0 pmol Leu-enkephalin equivalents per mg protein. The amount of opioids in the hybrid cells is strongly dependent on the cell density. The findings suggest that neuroblastoma X glioma hybrid cells contain opioid peptides that are very similar, if not identical, to Met- and Leu enkephalin. Opioid activity can also be detected in other neuronal cell lines and even in glioma cells. PMID- 6283024 TI - Postnatal changes in enzyme activities in synaptosomes isolated from hamster optic nerve endings. AB - Subsynaptosomal fractions isolated from optic terminal nuclei of adult and neonatal hamsters exhibited developmental changes in specific density, mitochondrial activity, and K+-stimulated, ouabain-inhibited p nitrophenylphosphatase (K-pNPPase) activity around the time of eye opening. The specific activity of K-pNPPase was six- to sevenfold higher after eye opening (14 16 days postnatal). A significant proportion of high-specific- activity K-pNPPase was recovered from the lightest subsynaptosomal fraction at all ages. This fraction contained very little external membrane by galactose oxidase - NaB3H4 labeling, suggesting that it may represent an internal pool, possibly the axonally transported form of the enzyme. Synaptic mitochondrial cytochrome c. oxidase activity also approximately doubled in the period between 12 and 16 days. The specific density of the external membrane increased very slowly, banding at 1.0 M sucrose at 12 and 16 days, and at 1.2 M in adults. These maturational events may reflect increased energetic needs for optic nerve endings following eye opening. PMID- 6283025 TI - Effects of cyclic AMP analogues and phosphodiesterase inhibitors on K+-induced [3H]noradrenaline release from rat brain slices and on its presynaptic alpha adrenergic modulation. AB - The possible role of cyclic AMP in the presynaptic alpha-adrenoceptor-mediated modulation of [3H]noradrenaline (NA) release induced by 13 mM K+ from superfused rat cerebral cortex slices was investigated. Both dibutyryl-cyclic AMP (db-cAMP) and 8-bromo-cyclic AMP (8-Br-cAMP) dose-dependently (10(-4) - 10(-2) M) enhanced K+-induced (3H]NA release, maximally to about 160% of control. In contrast, db cAMP had no effect on calcium-induced [3H]NA release in the presence of the calcium ionophore A 23187. Surprisingly, the phosphodiesterase (PDE) inhibitors 3 isobutyl-1-methylxanthine (IBMX). 7-benzyl-IBMX, 4-(3-cyclopentyloxy-4 methoxyphenyl)-2-pyrrolidone (ZK 62771), and 4-(3-butoxy-4-methoxybenzyl)-2 imidazolidinone (Ro 20-1724) appeared to inhibit K+-induced [3H]NA release in a dose-dependent (10(-5) - 10(-3) M) manner. At a concentration of 10(-4) M, AK 62771 caused an inhibition of [3H]NA release by 30%, and this inhibitory effect was not affected by 10(-6) M phentolamine nor by 10(-3) M db-cAMP or 10(-4) M theophylline. Theophylline by itself enhanced [3H]NA release to about 135% of control. The inhibitor effect of the alpha-adrenoceptor agonist oxymetazoline (1 micro M) and the enhancing effect of the antagonist phentolamine (1 micro M) on [3H]NA release were significantly decreased in the presence of 10(-3) M db-cAMP or 8-Br-cAMP, whereas 10(-4) M ZK 62771 had no effect. In the presence of 10(-2) M NaF, a potent activator of adenylate cyclase, the inhibitory effect of oxymetazoline (1 micro M) on [3H]NA release was significantly decreased. The data obtained with the cyclic AMP analogues support the hypothesis that activation of presynaptic alpha-receptors modulating NA release results in an inhibition of a presynaptic adenylate cyclase. Possible causes for the anomalous effects of th PDE inhibitors are discussed. PMID- 6283026 TI - Alterations of brain cholecystokinin receptors in mice made obese with goldthioglucose. PMID- 6283027 TI - Cyclic nucleotides and the release of vasopressin from the rat posterior pituitary gland. AB - When rat posterior pituitary glands were stimulated by a high concentration of potassium, a peak of cyclic AMP and a peak of cyclic GMP were detected after 0.5 min and 1 min, respectively, whereas the rate of release of vasopressin was maximal only after 2 min. When calcium was omitted from the medium, no significant changes in cyclic nucleotide levels were found and the vasopressin release remained at the basal rate. During cold-stimulated (10 degrees C) release of vasopressin, a peak of cyclic AMP was detected after 5 min simultaneously with the maximal rate of vasopressin release. The significance of the cyclic nucleotides in the release of vasopressin is discussed. PMID- 6283028 TI - Histocompatibility antigens on astrocytoma cells. AB - Biopsies tumour cells from astrocytoma-bearing patients were grown in primary culture for 3-5 days. Both low and high grade tumours were represented in the study. The cultured cells could be shown to express the HLA-A and -B antigens using a multispecific allo-antiserum and a rabbit anti-beta-2 microglobulin antibody. The tumour cells were negative for the HLA-DR determinants when tested with either rabbit anti-Ia-like antisera or specific anti-HLA-DR allo-antisera. They also failed to stimulate allogeneic lymphocytes in primary mixed lymphocyte tumour cell cultures but stimulated lymphocytes primed to tumour cells in vitro. The tumour cells were also capable of stimulating autologous lymphocytes from the tumour-bearing patient in most of the combinations tested. PMID- 6283029 TI - Ventilatory failure in myasthenia gravis. AB - This retrospective study over the decade 1969-1978 examines the precipitating factors and outcome in thirty-one patients with myasthenia gravis who developed ventilatory failure. An unusual example of chronic alveolar hypoventilation is discussed in detail. The most favourable outcome occurred in younger patients with a hyperplastic thymus, in contrast to a poorer outlook for older patients with an atrophic gland. Eleven patients died during the period of follow up: three deaths were unrelated to myasthenia but the remaining eight were attributed directly or indirectly to it. The mortality of 36% represents a marked improvement on a 70% mortality in a similar group of patients, reported from this hospital for the years 1960-1968. PMID- 6283030 TI - Diagnostic value of cerebrospinal fluid antibodies in herpes simplex virus encephalitis. AB - Antibodies to different viruses and bacteria were measured in the cerebrospinal fluid (CSF) of six patients with herpes simplex virus encephalitis proven by brain biopsy and in five others with a presumptive diagnosis. Antibodies to herpes simplex virus but not to other organisms appeared in the CSF of all patients after the first weeks of the illness. Herpes simplex virus antibodies were not found in control CSF. The antibodies persisted in the CSF and the serum/CSF antibody ratio remained altered, 32:1 to less than 1:1, in all cases during the follow-up to 29 months or until death. The CSF albumin level was normal and the IgG index (formula: see text) elevated in four proven and three presumptive cases indicating a local antibody production; in four patients the findings were inconsistent. These results suggest that prolonged antigen stimulation is present in the central nervous system after acute herpes simplex encephalitis and that serological measurements combined with immunoglobulin and albumin determinations may provide a tentative but not definite diagnosis in some cases after the acute phase of encephalitis together with a method for follow-up of patients. PMID- 6283031 TI - Conduction of sensory action potentials across the posterior fossa in infratentorial space-occupying lesions in man. AB - Central conduction time following median nerve stimulation has been recorded in 28 patients with infratentorial space occupying lesions. Fourteen of these (50%) showed a prolonged central conduction time, which invariably involved that segment of the conduction time between the dorsal column nuclei generators and the generation site for P15, which we believe to be the upper brain stem or thalamus. Three other cases showed prolongation of the N14-P15 segments though total central conduction time was within normal limits. One case with marked hydrocephalus showed prolongation both before and after the P15 wave. The possible pathophysiology of prolongation of central conduction time in these cases and the possible neural generation site of P15 are discussed. PMID- 6283032 TI - Sensory nerve conduction in the human spinal cord: epidural recordings made during scoliosis surgery. AB - This report describes the waveform and properties of somatosensory evoked potentials recorded from various levels of the human spinal cord, with electrodes inserted into the epidural space and the stimulus delivered to the posterior tibial nerve at the knee. The object was to provide a means of monitoring spinal cord function during surgery for the correction of spinal deformities. The responses could be resolved into at least three components with different activation thresholds and different conduction velocities within the spinal cord (45-80 m/s approximately). The findings are in accord with recent studies, suggesting that the fast activity may be conducted in the dorsal spinocerebellar tract and the slower waves in the posterior columns. PMID- 6283033 TI - Migraine patients exhibit abnormalities in the visual evoked potential. AB - An experiment is described which compared visual evoked potential (VEP) amplitudes and latencies in normal subjects and migraine patients. Several VEP abnormalities were found: at vertex and to a lesser extent at temporal sites, P100-N120 amplitudes were larger in patients; at vertex and temporal sites N120 amplitudes were larger in patients; at temporal sites patents had larger N120 P200 amplitudes but smaller P100 amplitudes. Peak latencies of the VEP were also found to be abnormal in patients. At vertex, patients had delayed N120 peak latencies while at temporal sites delays were found in the P200 latencies. Patients were subdivided according to side of headache. Right-sided headache patients showed larger temporal P100 amplitudes and larger left temporal P100 N120 amplitudes than bilateral headache patients. PMID- 6283034 TI - Peripheral neuropathy and solitary myeloma: analysis of serum and CSF IgG in two cases. AB - Clinical and laboratory findings of two patients affected by solitary myeloma, IgG3 lambda type, with peripheral neuropathy are reported. In both cases the same IgG isoelectrofocusing pattern was found in cerebrospinal fluid and serum samples. Data are consistent with damage of the blood-brain barrier. PMID- 6283035 TI - Reduced nerve blood flow in hexachlorophene neuropathy: relationship to elevated endoneurial fluid pressure. AB - To test the hypothesis that increased endoneurial fluid pressure (EFP) causes a reduction in nerve blood flow (NBF) in the vasa nervorum, we adapted a noninvasive method for measurement of nerve blood flow which was originally developed for measurement of local cerebral blood flow. This technique measures tissue distribution to the radioisotope, 14C-iodoantipyrine, and was used to compare NBF in sciatic nerves of rats with increased EFP induced by feeding them hexachlorophene (HCP), a neurotoxin which causes edema exclusively to the nervous system and confined to the myelin sheath. Elevation of interstitial fluid pressure in peripheral nerves from control values of 2.0 +/- 1.0 cm H2O to over approximately 6 cm H2O was associated with a statistically significant reduction in nerve blood flow from 14.8 +/- 5.9 to 7.8 +/- 2.5 ml/100 g of tissue/minute (min). These results support the hypothesis that increased endoneurial fluid pressure exacerbates the neuropathy by diminishing local blood flow. PMID- 6283036 TI - Cell body remodeling during dying-back axonopathy: DRG changes during advanced disease. AB - Although the neuronal cell body functions critically in maintaining and repairing the axon, little attention has been focused on the soma during neurotoxin-induced dying-back of the axon. To help elucidate changes in the perikarya during axonal dying-back, this study employed the hexacarbon model of axonal neuropathy in a setting designed to maximize the chances of detecting structural alterations. A spectrum of cell body modifications occurred in the fifth lumbar dorsal root ganglia (DRG) of rats chronically exposed to 2,5-hexanedione. These included distinctive cytoplasmic remodeling, a perineuronal cell reaction, and evidence of neuronal death with neuronophagia. These findings suggest the importance of the cell body in the pathogenesis of neurotoxin-induced axonal degeneration and imply that complex soma-axon interactions may help determine the fate of intoxicated neurons. PMID- 6283037 TI - Peripheral neuropathy associated with monoclonal gammopathy. Studies of intraneural injections of monoclonal immunoglobulin sera. AB - A causal relationship between paraproteinemia and neuropathies has been suggested. We studied three patients with chronic sensorimotor polyneuropathy associated with plasma cell dyscrasia and monoclonal gammopathies (IgGK, IgMK, IgA lambda). Sural nerve biopsies showed mild (2 cases) to moderate loss of myelinated fibers (1 case). Teased single fiber studies showed segmental demyelination-remyelination in two patients. Direct immunofluorescence demonstrated immune deposits of the myelin sheath of the same specificity as the serum paraprotein, IgGK (1 of 3 cases). Treatment with prednisone, melphalan or chlorambucil, and plasmapheresis resulted in remission (1 case), partial improvement (1 case), or had no effect (1 case), although reduction of monoclonal immunoglobulin occurred in all. To investigate the role the paraproteins might play in the pathogenesis of the neuropathy, patients' serum was injected intraneurally into rat sciatic nerves. None of the animals developed weakness, slowing of in vitro conduction of sciatic nerve, or significant evidence of demyelination by light- or electron-microscopy or teased single fiber studies 48 hours postinjection. Similar injections of rabbit serum with experimental allergic neuritis (EAN) produced focal segmental demyelination. Our studies employing an in vivo bioassay technique failed to establish antimyelin activity of monoclonal immunoglobulin sera. PMID- 6283038 TI - Abnormal proliferation of intraergastoplasmic microfilaments in myelinated schwann cells: ultrastructural study of two cases. AB - We report the ultrastructural findings in superficial peroneal nerve biopsies in two patients, one with idiopathic sensory neuropathy and the other with the Guillain-Barre syndrome and Hodgkin's disease. In addition to demyelination, there was an intense proliferation of microfilaments within numerous vesicles of the endoplasmic reticulum of a great number of myelinated Schwann cells. This abnormality does not appear to have been previously reported in the literature. The mechanisms responsible for this finding are unknown. PMID- 6283039 TI - The blocking action of baclofen on excitatory transmission in the rat hippocampal slice. AB - The mode of action of baclofen on the physiology of the rat hippocampus was investigated by studying its effect on electrophysiological responses in the hippocampal slice preparation and by measuring biochemical parameters related to glutamate uptake, binding, and release. Baclofen inhibited, in a dose-dependent fashion, the dendritic field potentials in field CA1 produced by stimulation of the Schaffer commissural fiber system. The drug was inactive in this respect at concentrations of 10 and 100 nM but consistently reduced the amplitude of both the dendritic field potential and the population spike at a concentration of 1 microM. At a concentration of 25 microM, baclofen virtually abolished the dendritic and cell body responses to afferent stimulation. Recovery of field potentials required between 7 and 10 min following the addition of 1 microM baclofen. The levorotatory form of baclofen was much more potent in suppressing synaptic responses than was the dextrorotatory enantiomer. Baclofen, at a concentration of 5 microM, strongly antagonized the excitation of pyramidal neurons evoked by iontophoretically applied glutamate. The antagonism of the glutamate effect was much reduced when the slices were maintained in low calcium, high magnesium perfusion medium. Moreover, under low calcium conditions, baclofen did not interfere with the effects of bath-applied glutamate on antidromically elicited responses. Baclofen did not affect the Na+-dependent or Na+-independent binding of [3H]glutamate to crude synaptic membrane fractions from the hippocampus. However, at a concentration of 1 microM, it markedly inhibited potassium-induced release of [3H]glutamate from hippocampal synaptosomes. Taken together, the present results strongly suggest that baclofen suppresses synaptic responses in the Schaffer commissural fiber system of the hippocampus by blocking the release of an excitatory amino acid transmitter. PMID- 6283040 TI - The binding of kappa- and sigma-opiates in rat brain. AB - Detailed displacements of [3H]dihydromorphine by ketocyclazocine and SKF 10,047, [3H]ethylketocyclazocine by SKF 10,047, and [3H]SKF 10,047 by ketocyclazocine are all multiphasic, suggesting multiple binding sites. After treating brain tissue in vitro with naloxazone, all displacements lose the initial inhibition of 3H ligand binding by low concentrations of unlabeled drugs. Together with Scatchard analysis of saturation experiments, these studies suggest a common site which binds mu-, kappa, and sigma-opiates and enkephalins equally well and with highest affinity (KD less than 1 nM). The ability of unlabeled drugs to displace the low affinity binding of [3H]dihydromorphine (KD = 3 nM), [3H]ethylketocyclazocine (KD = 4 nM), [3H]SKF 10,047 (KD = 6 nM), and D-Ala2-D-Leu5-[3H]enkephalin (KD = 5 nM) remaining after treating tissue with naloxazone demonstrates unique pharmacological profiles for each. These results suggest the existence of distinct binding sites for kappa- and sigma-opiates which differ from those sites which selectively bind morphine (mu) and enkephalin (delta). PMID- 6283041 TI - The effects of microstimulation and microlesions in the ventral and dorsal respiratory groups in medulla of cat. AB - The responses in respiratory outflow resulting from microstimulation and successive microlesions of the dorsal (DRG) and ventral (VRG) respiratory groups of neurons in the brainstem were studied in anesthetized, paralyzed, artificially ventilated cats. Microstimulation (2 to 120 Hz; 5 to 50 microA; 100 musec pulse duration) at almost every point within the DRG or VRG produced a bilateral short latency inhibition of phrenic nerve activity which had an onset latency of 4 to 9 msec and a duration of 4 to 24 msec. This global stereotyped phrenic inhibition was elicited by single pulses and often was accompanied by a postinhibitory excitation. In 48% (92/193) of the stimulation trials, trains of stimulus pulses during inspiration decreased the duration of inspiration. In 25% of the expiratory microstimulation trials, expiratory duration was increased and in 11%, expiration was shortened markedly by trains of pulses. Single shocks delivered to the right VRG or DRG produced a short latency excitation in the ipsilateral recurrent laryngeal nerve (RRL). This RRL excitation had an onset latency of 2 to 5 msec and a duration of 3 to 15 msec. Evidence suggests that the RRL excitation is due to a paucisynaptic activation of expiratory motoneurons in the caudal VRG. This activation is synchronous with the inhibition of inspiratory neurons in DRG and VRG. Despite the powerful short latency effects of microstimulation in VRG and DRG, extensive bilateral destruction of these neuronal populations had only modest effects on respiratory rhythm, while it decreased (or abolished) respiratory outflow in phrenic and recurrent laryngeal nerves. The combined results of the microstimulation and microlesion portions of this study suggest that a region (or regions) outside of the DRG and VRG might be important in the control of the respiratory pattern and that the DRG and VRG are important in determining the depth of inspiration; their role in generating respiratory rhythm needs to be critically re-examined. PMID- 6283042 TI - Effects of intravenous methylprednisolone on spinal cord lipid peroxidation and Na+ + K+)-ATPase activity. Dose-response analysis during 1st hour after contusion injury in the cat. PMID- 6283043 TI - Complication of spinal tap. PMID- 6283044 TI - Feeding responses of riboflavin-deficient rats to energy dilution, cold exposure and glucoprivation. AB - Feeding in response to various stimuli, was investigated in riboflavin-deficient rats. The animals failed to increase their food intake when fed energy-diluted diets with cellulose contents of either 15 or 30%. Furthermore, they lost reactivity to the hyperphagic action of either insulin of 2-deoxy-D-glucose regardless of a perfect response in blood glucose. Cold exposure (10 degrees), however, stimulated energy intake of the deficient rats to the same extent as that of the riboflavin-supplemented normal rats. These findings indicate that there was an operative thermostatic, but not a glucostatic control in regulation of energy intake of the riboflavin-deficient rats. PMID- 6283045 TI - Excretion of breath and flatus gases by humans consuming high-fiber diets. AB - The abilities of dietary fibers to promote the excretion of intestinal fermentation gases were evaluated in five healty men. Flatus and breath gases were collected and analyzed during 3 days of each 9-day metabolic period. Responses to feeding xylan, pectin, cellulose and corn bran were compared to a fiber-free formula diet. Generally, hydrogen production increased throughout the day, whereas methane production remained more constant. Methane excretion was greater while consuming the xylan and pectin diets than while consuming the other diets. These two purified fibers also caused higher flatus volume, hydrogen and carbon dioxide excretion. Cellulose and corn bran generally resulted in breath and flatus gas excretion at levels equivalent to fiber-free diets. Considerable variation was noted in the response of subjects to an individual diet. However, in most cases 2-5 days of frequent and daily consumption of the diets was necessary to establish a relatively constant level of gas excretion. This time may represent the period of microbial and enzymatic adaptation to the new dietary constituents. PMID- 6283046 TI - Lingual osteoma. PMID- 6283047 TI - [Malignant lymphoma of the nasopharynx of non-Burkitt's type--a case report of a patient with positive Epstein-Barr virus-associated nuclear antigen (EBNA) (author's transl)]. PMID- 6283048 TI - Inclusion body fibromatosis. AB - The clinical and pathological features of two further cases of the condition previously described as Recurring Digital Fibrous Tumour of Childhood are presented. The ultrastructural appearance of the characteristic cell inclusions bears some resemblance to Mallory's hyaline and suggest increased functional activities in the fibroblasts carrying these unusual dense bodies. Retention of abnormal metabolic products in fibroblasts is the most likely explanation. The term Inclusion Body Fibromatosis is proposed for this entity. PMID- 6283049 TI - Genetic diseases: diagnosis by restriction endonuclease analysis. AB - We have summarized a number of different genetic disorders which can be diagnosed at the DNA level using restriction endonuclease fragment analysis. A whole spectrum of defects can be recognized: point mutations, deletions, additions, and crossing-over products or hybrid genes. These same restriction endonuclease techniques can enable different genes to be marked by polymorphism patterns. Thus, abnormal genes can be identified even if their exact DNA lesion is unknown or cannot be directly detected. The progress that has been made with the hemoglobinopathies and the experience from this group of single gene disorders should find application to other diseases as soon as specific probes become available. PMID- 6283050 TI - Immunologic and epidemiologic aspects of varicella infection acquired during infancy and early childhood. AB - The development of varicella zoster infection was studied in a population of infants under one year of age during three outbreaks of varicella in a semi closed domiciliary institution for infants in Japan. Over a period of four years, many residents ranging in age from 27 days to 32 months were tested for cutaneous reactivity to VZV antigen, and VZV-specific antibody activity before, during, and after each outbreak of varicella. Of these, 85 subjects developed clinical varicella, with an overall attack rate of 100% for all susceptible subjects. All the infants under 2 months of age were infected following such exposure, despite the presence of pre-existing maternal antibody. The degree of cutaneous involvement appeared to be milder (less than 20 vesicles) in infants less than 2 months of age, and severe cutaneous disease (with over 300 eruptions or confluent rash) occurred more frequently in subjects 2 to 11 months of age. Pre-existing antibody did not prevent development of illness, or alter the degree of antibody or cellular immune response to subsequent infection. However, the peak cutaneous reactivity to VZV antigen after infection was found to be significantly lower in infants under 2 months of age. PMID- 6283051 TI - Live attenuated varicella vaccine: immunization of healthy children with the OKA strain. AB - Ninety-one healthy children were immunized with one of three preparations of the OKA strain of live attenuated varicella zoster virus in order to gain sufficient information on safety and immunogenicity to justify vaccine studies in immunosuppressed children. The OKA-Biken vaccine was studied in a dose response trial. Two additional clinical trails were conducted with the OKA strain that was further passaged: a frozen aqueous (OKA-RIT-Aq) form and a lyophilized (OKA-RIT Ly) form were tested. With each of these vaccines clinical reactivity was minimal and virus shedding, viremia, and transmission of the vaccine virus were absent. Excellent seroconversion rates of 100 and 94% were induced with approximately 500 pfu of OKA-Biken and OKA-RIT-Ly vaccine, respectively. A lower seroconversion rate was noted for OKA-RIT-Aq, presumably on the basis of defective individual vials. Diluted OKA-Biken vaccine induced less seroconversion but even the 5 pfu dose produced antibody responses in 81% of the vaccinees. In vitro specific cellular immune responses to varicella virus antigens were demonstrated in each group of vaccinees, including all but two of those vaccines who failed to seroconvert. The data indicate that two preparations of the OKA strain are sufficiently safe and immunogenic in healthy children to support further studies in immunocompromised children. PMID- 6283052 TI - The role of respiratory syncytial virus and other viral pathogens in acute otitis media. AB - We utilized recently developed enzyme immunoassay techniques to examine the role of selected viruses in the etiology of acute otitis media. Viral pathogens were found in middle ear fluids obtained from 13 (24%) of 53 children with acute otitis media; respiratory syncytial virus accounted for ten of the 13 viral agents identified. In addition, respiratory syncytial viral antigen was found in nasopharyngeal washings obtained from 15 of the 53 children. Seven of these children had RSV identified as the sole middle ear pathogen, whereas six children had otitis caused by Streptococcus pneumoniae as either the sole middle ear pathogen or in combination with RSV. Similarly, all three children with respiratory infections caused by influenza virus had ear infections caused by bacterial pathogens, either alone or in combination with influenza virus. These findings suggest that, in patients with viral respiratory infection, coexisting acute otitis media may be associated with the recovery of either viruses or bacteria from the middle ear exudates. PMID- 6283053 TI - Gastroenteritis associated with enteric type adenovirus in hospitalized infants. AB - Enteric types of adenovirus have recently been identified as a causative agent of infantile gastroenteritis. We utilized enzyme immunoassay and tissue culture techniques to evaluate prospectively the role of ET Ad in diarrhea occurring in hospitalized infants. We found that ET Ad was associated with 14 of 27 cases of diarrhea occurring during a 12-week study period in the late autumn and early winter months; ET Ad was found in the stool of only one of 72 children without diarrhea (P less than 0.001). Although adenoviruses other than ET Ad were found in the stools of two of the 27 children with diarrhea, such viruses were also found in the stools of five of 72 children without diarrhea and thus could not be statistically correlated with acute gastroenteritis. Children infected with ET Ad had diarrhea for a mean of 8.0 days, compared to a mean duration of 4.2 days for the children with gastroenteritis not associated with ET Ad. Thirteen of the 14 children with ET Ad gastroenteritis had respiratory symptoms such as cough, rhinorrhea, or wheezing, six had roentgenographic evidence of pneumonia, and three children had bilateral conjunctivitis. This study documents that ET Ad can be an important cause of acute gastrointestinal disease in hospitalized infants and young children and that gastrointestinal infections with ET Ad can be associated with a high rate of respiratory disease. PMID- 6283054 TI - Nonfebrile seizures after febrile convulsions: possible role of chronic cytomegalovirus infection. AB - Twelve hundred children with convulsions when feverish were studied during a period of five years. Among them 52 subjects (4.33%) developed nonfebrile seizures after a period of eight months to five years from the first febrile convulsion (group A). Twenty-three children had neither afebrile seizures nor EEG abnormalities during the period of observation (group B). The two groups were comparable for age of the first febrile convulsion onset, sex, and socioeconomic status. None had risk factors for subsequent epilepsy or clinical signs of congenital cytomegalovirus infection. The isolation rate of CMV from urine was 53.84% in patients of group A, 26.09% in children of group B, and 26.83% in healthy control children. Twelve CMV-positive children from group A were followed for one to more than three years. In five of seven children with persisting EEG abnormalities, cytomegaloviruria was still present 13 to 41 months after the first isolation, whereas none of five patients with normal electroencephalograms had viruria after a comparable period. We found that CMV-positive children generally lacked cell-mediated immunity to the virus, whereas CMV-negative patients had positive reactions. Our data suggest a correlation between persistence of neurologic abnormalities and CMV excretion in children with nonfebrile seizures and CMV infection. PMID- 6283055 TI - Hepatoma in an HBsAg carrier--seven years after perinatal infection. PMID- 6283056 TI - Vascular tolerance to nitroglycerin and cyclic GMP generation in rat aortic smooth muscle. AB - Recent reports have suggested that cyclic guanosine 3', 5'-monophosphate (cGMP) may mediate drug-induced vascular smooth muscle relaxation. This hypothesis was examined by correlating increases in cGMP formation with vascular smooth muscle relaxation after sensitivities to nitroglycerin and nitroprusside were altered. Both nitroglycerin and nitroprusside produced a concentration-dependent increase in rat aortic cGMP levels, which preceded relaxation. A specific inhibition of nitroglycerin relaxation was accomplished by pretreating helical rat aortic strips with 5.5 X 10(-4) M nitroglycerin for 1 hr (in vitro nitroglycerin tolerance). Both nitroglycerin and nitroprusside were inhibited by pretreating tissues with 10(-5) M methylene blue for 1 hr. Whenever nitroglycerin or nitroprusside relaxation was inhibited, there was a corresponding inability to generate cGMP. Vascular tissues derived from nitroglycerin-tolerant animals (in vivo tolerance) exhibited a profile of cGMP reduction that closely resembled that of the in vitro model. Vascular smooth muscle relaxation induced by 8 bromoguanosine-3', 5'-monophosphoric acid was not inhibited by either in vitro nitroglycerin tolerance or methylene blue, suggesting that the subsequent action of cGMP, once formed, is not impaired. The results of this study support the contention that cGMP generation may be an important step in the promotion of vascular smooth muscle relaxation by nitroglycerin and nitroprusside. PMID- 6283057 TI - Comparison of the effects of trimethadione and its primary metabolite dimethadione on neuromuscular function and the effects of altered pH on the actions of dimethadione. AB - The effects of the anticonvulsant trimethadione (TMO) and its primary metabolite dimethadione (DMO) were investigated at the frog neuromuscular junction using intracellular recording techniques. TMO (1, 2 and 5 mM) caused dose-dependent decreases in miniature end-plate potential (MEPP) and end-plate potential (EPP) amplitudes, decreased quantal content only at the highest dose and did not affect MEPP frequency. DMO (2 and 5 mM) at the normal pH of 7.2 significantly decreased quantal content and decreased EPP amplitude at the higher concentration used. Neither MEPP amplitude nor frequency was significantly affected by DMO at pH 7.2. When 2 mM DMO was added at the same time that pH was lowered to 6.6, considerably larger decreases in EPP( amplitude and quantal content were observed. Under these conditions, DMO still did not alter MEPP amplitude but did cause about a doubling in MEPP frequency. The effects of pH 6.6 alone were also examined, but lowered pH did not account for all of the exaggerated effect of DMO in pH 6.6. Presumably, more DMO accumulates intracellularly in low pH conditions because it is a weak acid and sensitive to alterations in pH. In conclusion, both TMO and DMO cause depression of neuromuscular transmission; however, their mechanisms for depression are different, especially when therapeutically relevant concentrations are considered. TMO acts primarily by suppressing postjunctional sensitivity to acetylcholine, whereas DMO primarily decreases transmitter release from the nerve terminal. PMID- 6283058 TI - Binding of a radiolabeled triazolopyridazine to a subtype of benzodiazepine receptor in the rat cerebellum. AB - The triazolopyridazine (TPZ) drugs, typified by CL218,872 (CL), have a relatively higher affinity for a subpopulation of benzodiazepine (BZ) receptors. The binding of radiolabeled CL to membranes from rat cerebellum, a region enriched in the TPZ preferring ("Type 1") BZ receptor, was characterized and compared with that of [3H]flunitrazepam ([3H]FLU) in the same preparation. [3H]CL had clonazepam displaceable binding which was saturable. The Kd was approximately 21 nM and the Bmax was approximately 600 fmol/mg of protein. [3H]CL binding was similar to that for [3H]FLU in that exogenous gamma-aminobutyric acid (GABA) enhanced the binding; however, [3H]CL binding differed from that for [3H]FLU in that anions, cartazolate and pentobarbital did not enhance [3H]CL binding. These data suggest that [3H]CL binds to the Type 1 BZ receptor in a manner different from that of a BZ drug such as FLU. Inasmuch as GABA enhances [3H]CL binding, but anions, cartazolate and pentobarbital do not, [3H]CL may bind to the Type 1 BZ receptor in such a way that it interacts with the GABA site, but perhaps not directly with the ionophore or the postulated pyrazolopyridine-barbiturate site. Thus, TPZ drugs may affect the GABA receptor complex in a different or perhaps less extensive way than the BZs. This, in addition to the regional localization of the Type 1 receptor, may be an important part of the mechanism of action of the TPZs. PMID- 6283060 TI - Evidence that the discriminative stimulus properties of fentanyl and ethylketocyclazocine are mediated by an interaction with different opiate receptors. PMID- 6283059 TI - Effects of MK-196 and furosemide on rat medullary thick ascending limbs of Henle in vitro. AB - The effects of the diuretic agents, MK-196 (6,7-dichloro-2-methyl-1-oxo-2-phenyl 5-indanyloxy acetic acid) and furosemide on function of medullary thick ascending limb (tALH) isolated from the rat kidney were studied. tALH were dissected from whole kidneys of male Sprague-Dawley rats which were first perfused in vitro in a recirculating system for 10 to 15 min with a solution containing collagenase. Standard techniques for dissection and perfusion of tALH were followed. Control transepithelial voltage averaged +5.5 +/- 0.5 mV (n = 25). Drug concentration causing 50% inhibition of the voltage (EC50) was determined. EC50 for both MK-196 and furosemide in rat tALH was 8 X 10(-6) M (EC50 for MK-196 in rabbit tALH ws 7 X 10(-5) M). In other experiments, net chloride transport (JCl) in rat tALH averaged 142 +/- 19 pmol.mm-1.min-1 (n = 12), a value nearly twice that reported for the rabbit. In the rat, MK-196 (2 X 10(-4) M) reduced the voltage by 79 +/- 2% and JCl by 76 +/- 13% (n = 6). The same concentration of furosemide inhibited the voltage by 78 +/- 8% and the JCl by 104 +/- 4%(n = 5). Results indicate that: 1) rat tALHs dissected from collagenase-treated kidneys have functional characteristics similar to those reported for the rabbit tALH; and 2) the rat tALH is equally sensitive to MK-196 and furosemide, drugs which act to selectively inhibit active chloride transport. PMID- 6283061 TI - Mode of the ciguatoxin-induced supersensitivity in the guinea-pig vas deferens. AB - Ciguatoxin (CTX; 10(-7) X 10(-7) g/ml), the most potent marine toxin isolated from a number of tropical and subtropical fishes, shifted the dose-contractile response curves for norepinephrine (NE) and K+ to the left in a parallel manner in the guinea-pig isolated vas deferens, indicating that CTX caused supersensitivity. The CTX-induced potentiation was inhibited or abolished in the presence of tetrodotoxin (5 X 10(-7) M) or saxitoxin (5 X 10(-7) M) and in Na+ deficient medium, but was not affected by phentolamine (10(-6) M) and verapamil (10(-6) M). Treatment with reserpine (2 mg/kg/day, twice) almost completely prevented the release of NE by CTS, such pretreatment had no affect on the ability of CTX to potentiate responses to NE and K+. Furthermore, after cold storage (4 degrees C for 7 days) of tissues, the contractile response to NE (3 X 10(-6) M) and K+ (20 mM) was still profoundly potentiated after treatment with CTX (5 X 10(-7) g/ml). CTX (10(-7)-10(-5) g/ml) by itself had no apparent effect on either Na+, K+-adenosine triphosphatase activity or Na+ content of the vas deferens. However, in the presence of ouabain, CTX elevated the Na content of the vas deferens treated with ouabain alone by 27%. This effect of CTX was abolished by tetrodotoxin. These data suggest that CTX causes an increasing Na+ permeability across the TTX sensitive Na+ channels of smooth muscle cell, and this may play an important role in its mechanism of potentiation. PMID- 6283062 TI - Extrasynaptic location of alpha-2 and noninnervated beta-2 adrenoceptors in the vascular system of the pithed normotensive rat. PMID- 6283063 TI - Positive inotropic effect of amrinone in relation to cyclic nucleotide metabolism in the canine ventricular muscle. AB - Experiments were carried out in the canine isolated right ventricular muscle in order to study the positive inotropic effect of amrinone in relation to the cyclic AMP and cyclic GMP metabolism. The positive inotropic effect of amrinone was accompanied by accumulation of cyclic AMP in the tissue: the cyclic AMP level was elevated in a similar time course to that of the increase in force of contraction and in a concentration-dependent manner in the presence of a beta adrenoceptor blocking agent, pindolol (3 X 10(-8) M). An excellent correlation was found between the force of contraction and cyclic AMP level in the presence of amrinone: the relationship was similar to that obtained previously with papaverine, a cyclic AMP phosphodiesterase inhibitor. The cyclic GMP level determined simultaneously was not changed by amrinone. The amrinone-induced increases in force of contraction and cyclic AMP level were markedly depressed by carbachol (3 X 10(-6) M); the relationship between the force of contraction and cyclic AMP level in the presence of amrinone was not changed by carbachol. Amrinone inhibited the cyclic AMP phosphodiesterase purified from the canine right ventricular muscle and enhanced the positive inotropic effect of isoproterenol. Amrinone shortened the time required for relaxation of a single contraction to the same extent as isoproterenol and cyclic AMP phosphodiesterase inhibitors such as 1-methyl-3-isobutylxanthine and papaverine. These results altogether strongly suggest that the change in cyclic AMP metabolism is involved in the positive inotropic effect of amrinone on the canine ventricular muscle. PMID- 6283064 TI - Selective desensitization of cardiac beta adrenoceptors by prolonged in vivo infusion of catecholamines in rats. AB - The effects of prolonged in vivo infusion of isoproterenol (400 micrograms/kg/hr) or norepinephrine (200 micrograms/kg/hr) from a minipump on the physiological reactivity and binding properties of cardiac beta and alpha-1 adrenoceptors were tested in rats. Infusion of either catecholamine significantly reduced the in vitro inotropic and chronotropic potency of isoproterenol in isolated left and right atria, respectively; desensitization was near maximal as early as after 2 hr of infusion. No significant change in the density of [3H]dihydroalprenolol labeled beta receptors was evident at this time point in either atrial or ventricular tissue, although isoproterenol did decrease binding site density after 7 days of infusion. There was no change in the binding affinity or physiological blocking potency of dihydroalprenolol after isoproterenol infusion. The inotropic potency of phenylephrine in the presence of dihydroalprenolol was unaffected by infusion of either isoproterenol or norepinephrine and methoxamine failed to increase right atrial rate either in control or in isoproterenol infused rats. There was also no change in the density and affinity of [3H]prazosin binding sites after isoproterenol infusion. These results indicate selective desensitization of cardiac beta receptors without changes in alpha-1 receptors by prolonged in vivo stimulation with catecholamines. This reaction pattern is different from the well documented effects of hypothyroidism, which include decreased sensitivity of cardiac beta and increased sensitivity of cardiac alpha-1 receptor-mediated responses in rats. Thus, the mechanisms responsible for altered receptor function in the two conditions appear to be different. PMID- 6283065 TI - Adrenergic control of immunoreactive beta-endorphin release from the pituitary of the rat: in vitro and in vivo studies. AB - The adrenergic control of pituitary beta-endorphin release was investigated by testing the effect of various alpha and beta adrenergic drugs on the secretion beta-endorphin-like immuno-reactivity (beta-END-LI) from primary cell cultures of anterior and neurointermediate lobes as well as into plasma in vivo. Incubation of anterior lobe cells with several adrenergic agonists (10-7-10-5 M) evoked significant release of beta-END-LI with the following potency order; l epinephrine greater than alpha-methylnorepinephrine greater than guanfacine = methoxamine = l-isoproterenol. The epinephrine- and isoproterenol-induced release from anterior lobes were both inhibited by phenoxybenzamine but not propranolol, suggesting mediation by alpha adrenoceptors. Release of beta-END-LI from cultured neurointermediate lobe cells was also stimulated 3-fold by epinephrine or isoproterenol (10-100 nM) but appeared to do so via a beta adrenergic mechanism because propranolol blocked their effects. Administration of l-epinephrine (100 micrograms/kg s.c.) or l-isoproterenol (200 micrograms/kg s.c.) to rats evoked a 3- to 4-fold increase in plasma beta-END-LI, which peaked 30 min after injection and was dose-dependent up to 500 micrograms/kg, Pretreatment with phenoxybenzamine (5 mg/kg i.p.) 30 min before epinephrine (100 micrograms/kg s.c.) inhibited the drug-induced rise whereas propranolol (2 mg/Kg i.p.) blocked the increase in plasma beta-END-LI after isoproterenol (200 micrograms/kg, s.c.) These results demonstrate that alpha and beta adrenergic stimulation selectively release beta-END-LI from cultured anterior and neurointermediate lobes, respectively. Furthermore, both mechanisms appear to be active in vivo. PMID- 6283066 TI - Alterations in opiate receptor function after chronic ethanol exposure. AB - The dose-response curve for morphine-induced stimulation of striatal dopamine metabolism was shifted to the right in mice which had been withdrawn for 24 hours after chronic consumption of an ethanol-containing liquid diet. The apparent ED50 for morphine was increased by 33% in ethanol-treated mice. Concomitant with the shift in the dose-response curve, the affinity for dihydromorphine of the high affinity caudate morphine receptor was decreased in ethanol-treated mice. The change in receptor properties after ethanol treatment included a decreased sensitivity of the receptor to the effects of sodium ion on morphine binding. The results suggest: 1) that the effect of morphine on dopamine metabolism in the mouse striatum is, at least in part, mediated by receptors that exhibit a high affinity for dihydromorphine: and 2) that ethanol treatment and withdrawal may induce specific changes in these particular opiate receptors. PMID- 6283067 TI - The affinity of morphine for its pharmacologic receptor in vivo. AB - Morphine receptor affinity, an important constant in the classification of opiate receptors, has not yet been determined by a pharmacologic method. In the present study, we used a standard pharmacologic procedure, that of partial receptor occlusion, to make this determination in the rat utilizing the hot water (55 degrees C) tail flick test. The occluding compound was buprenorphine, a new narcotic antagonist analgesic that can bind to opiate receptors in an irreversible manner. When buprenorphine was given to rats in doses of 0.30 and 0.60 mg/kg (s.c.), 30 min before morphine (20-320 mg/kg s.c.) and 60 min before testing, it produced the theoretically predicted alterations in the morphine dose response relation that are indicative of partial receptor blockade. The morphine receptor apparent dissociation constant (reciprocal of affinity) was calculated to be 1.7 X 10-6 M. This value is greater than those previously obtained in radiolabeled binding studies which have ranged from 3 to 27 X 10-9 M in the absence of sodium chloride to 1 to 5 X 10-7 M in the presence of sodium chloride. Used in this way, buprenorphine may be a valuable tool in the determination of apparent dissociation constants for other narcotic agonists. PMID- 6283068 TI - In vitro and in vivo mechanisms of leukotriene-mediated bronchoconstriction in the guinea pig. PMID- 6283069 TI - Involvement of alpha noradrenergic receptors in mediation of brain polyphosphoinositides metabolism in vivo. AB - The acute effects of norepinephrine (NE) on polyphosphoinositide (PPI) metabolism were examined in regions of rat brain in vivo with the techniques of intracisternal injection and microwave irradiation fixation. NE (80 nmol/g of brain) increased PPI incorporation of both [32P] orthophosphate and [3H] inositol by 50 to 100% within 5 min of the simultaneous intracisternal administration of both drug and radiolabel. These effects of NE were more pronounced in the brainstem and cerebellum than in the cerebral cortex, and were greater for triphosphoinositide than for diphosphoinositide. The increases were blocked by phentolamine and phenoxybenzamine but not by propranolol or atropine, suggesting that alpha noradrenergic receptors mediated the effect. Similar metabolic responses were observed with PPI precursors, phosphatidic acid and monophosphoinositide. However, no other phospholipid was altered in response to the agents administered in these studies (data not shown). Increased incorporation of radiolabels into PPI was never accompanied by detectable increases in endogenous PPI concentrations, suggesting that head group turnover was stimulated rather than whole molecule synthesis. Also, the increase in PPI phosphate radiolabel incorporation was not due to NE effects on the specific activity of phosphate donor precursors (nucleotides). The present data suggest that the PPI may be involved in the membrane processes that result from stimulation of alpha noradrenergic receptors in brain. PMID- 6283070 TI - Characterization of the antihypertensive properties of ketanserin (R 41 468) in rats. PMID- 6283071 TI - A norepinephrine-cyclic AMP link in the hypothalamic pathways which mediate fever induced by endotoxin and prostaglandin E2 in the rat. AB - The changes in rectal temperature, metabolic rate, cutaneous temperatures and respiratory evaporative heat loss produced by an injection of a bacterial endotoxin piromen (4-40 ng in 1 microliter) into the anterior hypothalamus were assessed in conscious rats in both sexes from a wide range of body mass and at various ambient temperatures (TaS). Intrahypothalamic injection of piromen increased metabolism and decreased cutaneous temperatures which led to fever in rats at Ta 8-30 degrees C. The monophasic fever was not significantly correlated with either body mass, sex difference of the Ta at which the experiments are carried out. Furthermore, daily intrahypothalamic injections of piromen produced no pyrogenic tolerance. Intrahypothalamic injections of either prostaglandin E2, norepinephrine, aminophylline or dibutyryl cyclic AMP also produced increased metabolism and decreased cutaneous temperature which led to fever at Ta 8-30 degrees C. In addition, the fever induced by intrahypothalamic injections of piromen, prostaglandin E2 or nor-epinephrine was greatly antagonized by pretreatment with intrahypothalamic injections of alpha or beta adrenergic antagonist. However, the fever induced by dibutyryl cyclic AMP or aminophylline was not affected by pretreatment with adrenergic receptor blockade. The data indicate that a norepinephrine- cyclic AMP link occurs in the hypothalamic pathways which mediate the piromen-induced or the prostaglandin E2-induced fever in rats. PMID- 6283072 TI - The interaction between aminoquinolines and presynaptically acting neurotoxins. AB - Chloroquine and hydroxychloroquine block neuromuscular transmission in isolated tissues from mouse, rat, guinea pig and chick. Blockade is associated with depressed muscle responses to potassium and abolished muscle responses to nicotinic cholinergic agonists. Within certain time and concentration limits, the blocking effects of chloroquine and hydroxychloroquine are reversible. Both drugs antagonize the onset of paralysis caused by botulinum neurotoxin types A and B, but neither drug antagonizes tetanus toxin or beta-bungarotoxin. The ability of chloroquine and hydroxychloroquine to antagonize botulinum toxin is not due to blockade of nicotinic cholinergic receptors. At concentrations that produce neuromuscular blockade, d-tubocurarine does not antagonize botulinum toxin types A and B. Chloroquine causes botulinum toxin to remain at an antitoxin sensitive site. These data could mean that chloroquine acts at the cell membrane to inhibit toxin binding or internalization, or that it acts in the cell interior to inhibit lysosomal processing of toxin. Whatever its action, chloroquine is the most effective antagonist of botulinum toxin yet described. PMID- 6283073 TI - Adenylate cyclase and beta adrenergic receptor development in the mouse heart. AB - We have demonstrated previously a postnatal peak for the beta adrenergic receptor in the heart and detected the appearance of a beta adrenergic receptor before an (-)-isoproterenol inducible increase in heart rate. The present study examined 1) agonist displaceable [3H] dihydroalprenolol (DHA) binding in the neonatal and adult mouse heart and 2) adenylate cyclase in fetal, neonatal and adult mouse heart. 3[H]DHA binding displaceable by (-)-isoproterenol gave a similar Ki from 1 day neonate through adult. Similar to the result found for antagonist displacement binding, there was a dramatic increase in the agonist displaceable [3H] DHA binding postnatally. The maximum was achieved in 2 weeks and then gradually declined to adult level. Cyclase activity (basal, (-)-isoproterenol- and NaF- stimulated) paralleled beta adrenergic receptor increases before birth. However, no early postnatal peak was present. In the 13 day fetal mouse heart, there is no (-)-isoproterenol increase in heart rate, but beta adrenergic receptor (13 +/- 4% of adult) and (-)-isoproterenol-stimulated adenylate cyclase activity (15 +/- 5% of adult) are present. It is concluded that 1) no significant difference exists between the agonist and antagonist displaceable [3H] DHA binding during development, 2) adenylate cyclase activity increases significantly during the last third of pregnancy in parallel with the beta adrenergic receptor, 3) both the beta adrenergic receptor and adenylate cyclase activity can be detected before the heart rate responses and 4) total adenylate cyclase activity does not increase in parallel with the early postnatal beta adrenergic receptor peak. PMID- 6283074 TI - Electrophoretic isoenzyme patterns of Entamoeba histolytica and Entamoeba chattoni in a primate survey. AB - Stocks of Entamoeba histolytica grown in a monoxenic culture system from the feces of nonhuman primates are compared with the eleven zymodemes of E. histolytica so far demonstrated from man. In a similar fashion, Entamoeba chattoni has also been grown and identified. Both E. histolytica and E. chattoni have been demonstrated in keepers of the primate collections. Comparisons have been made using the electrophoretic patterns of three enzymes: glucosephosphate isomerase [(GPI) E.C.5.3.1.9], phosphoglucomutase [(PGM) E.C.2.7.5.1], and L malate--NADP+ oxidoreductase (oxaloacetate-decarboxylating) [(ME) E.C.1.1.1.40]. Enzyme patterns of E. histolytica from the apes were found to be identical with three of those already demonstrated from man. The enzyme pattern of E. chattoni was distinctly different from that of any of the E. histolytica zymodemes. Other protozoa found in the single fecal sample examined from each subject are also listed. PMID- 6283075 TI - Atypical glomus tumours. PMID- 6283076 TI - Anterior-pituitary adrenal function of gold-treated patients with juvenile rheumatoid arthritis. AB - The hypothesis that gold therapy enhances endogenous cortisol secretion was tested in juvenile rheumatoid arthritis patients. The secretion of cortisol, dehydroepiandrosterone-sulfate (DHEA-S) and several peptide hormones was determined from plasma samples collected every 20 min for 24 hours. In random patients cortisol secretion was significantly greater in gold treated patients than in similar patients not receiving gold. However, when untreated patients were restudied after initiation of gold therapy, there was no significant change in cortisol secretion. Gold therapy also did not significantly alter secretion of the peptide hormones or DHEA-S. Thus, gold does not appear to influence endogenous hormone secretion. PMID- 6283077 TI - New prospects for control of virus diarrhoea in children. PMID- 6283078 TI - The bursa: Fabricius to Delaware. PMID- 6283079 TI - Post-viral haemophagocytic syndrome. PMID- 6283080 TI - Renal transplantation in Anderson-Fabry disease. PMID- 6283081 TI - Homosexual transmission of amoebiasis. PMID- 6283082 TI - Stereochemical studies on medicinal agents. 25. Absolute configuration and analgetic potency of beta-1,2-dimethyl-2-phenyl-4-(propionyloxy)piperidine enantiomers. AB - Enantiomers of beta-1,2-dimethyl-4-phenyl-4-(propionyloxy)piperidine (4) were employed as probes to demonstrate that opioid receptors are capable of distinguishing between the enantiotopic edges (the Ogston effect) of the piperidine ring. These enantiomers, (-)- and (+)-4.HCl, were prepared by esterification of the corresponding alcohols, (+)- and (-)-4a. Single crystal X ray studies of (-)-4a.HCl reveal that it possesses the 2R,4S absolute configuration. Analgetic testing in mice (hot-plate) and receptor binding studies indicate that (-)-(2S,4R)-4.HCl is approximately ten times more potent than its enantiomer. The results are consistent with the operation of the Ogston effect in the interaction of achiral 4-phenylpiperidines with opioid receptors. Additionally, it is suggested that the piperidine ring of these and other closely related 4-phenylpiperidines bind within a receptor subsite cleft whose dimensions exclude diequatorial 2,6- and 3,5-dimethyl-substituted ligands. PMID- 6283083 TI - 1-beta-D-arabinofuranosyl-1H-imidazo[4,5-c]pyridine (ara-3-deazaadenine). PMID- 6283084 TI - Acute gastric dilation and rupture in Macaca arctoides associated with Clostridium perfringens. AB - A clinical case of gastric dilation and rupture is described in an adult male Macaca arctoides. Clostridium perfringens was isolated from the heart blood and liver. Data collected from our macaque colony for 13 years indicated a mortality rate of 3.5% due to acute dilation. PMID- 6283085 TI - Molecular dynamics of ferrocytochrome c. Magnitude and anisotropy of atomic displacements. PMID- 6283086 TI - Alkaline isomerization of ferricytochrome c: lysine is not replacing methionine at the sixth co-ordination site of the haem iron. PMID- 6283087 TI - An alternative view of mammalian DNA sequence organization. II. Short repetitive sequences are organized into scrambled tandem clusters in Syrian hamster DNA. PMID- 6283088 TI - Nucleotide sequence of terminal repeats of 412 transposable elements of Drosophila melanogaster. A similarity to proviral long terminal repeats and its implications for the mechanism of transposition. PMID- 6283089 TI - Bacteriophage P22 mutants that alter the specificity of DNA packaging. PMID- 6283090 TI - Maturation cleavage of bacteriophage P22 DNA in the absence of DNA packaging. PMID- 6283092 TI - Steps in the reaction mechanism of the Haemophilus influenzae Rf restriction endonuclease. PMID- 6283091 TI - Transcript maps of Drosophila yolk protein genes. PMID- 6283093 TI - Is the amino acid but not the nucleotide sequence of the Escherichia coli araC gene conserved? PMID- 6283094 TI - The gol site: a cis-acting bacteriophage T4 regulatory region that can affect expression of all the T4 late genes. PMID- 6283095 TI - Two species of chromatin-RNA polymerase II complex released from rat liver nuclei by nuclease digestion. PMID- 6283096 TI - Theoretical aspects of specific and non-specific equilibrium binding of proteins to DNA as studied by the nitrocellulose filter binding assay. Co-operative and non-co-operative binding to a one-dimensional lattice. PMID- 6283097 TI - Isolation of a multigene family containing human alpha-tubulin sequences. PMID- 6283098 TI - Cruciform structures in palindromic DNA are favored by DNA supercoiling. PMID- 6283099 TI - Nucleotide sequence of the trpD gene, encoding anthranilate synthetase component II of Escherichia coli. PMID- 6283100 TI - Delta sequences and double symmetry in a yeast chromosomal replicator region. PMID- 6283101 TI - Structure of cytochrome c551 from Pseudomonas aeruginosa refined at 1.6 A resolution and comparison of the two redox forms. PMID- 6283102 TI - Electrical and contractile consequences of Na+ or Ca2+ gradient reduction in cultured heart cells. PMID- 6283103 TI - A biochemical and ultrastructural study on the endogenous protein kinase activity of secretory granule membranes of prostatic origin in human seminal plasma. PMID- 6283104 TI - Energy-dependent transformation of mouse gall bladder epithelial cells in a Ca2+ depleted medium. PMID- 6283105 TI - Evidence the pp60src, the product of the Rous sarcoma virus src gene, undergoes autophosphorylation. AB - pp60src, the product of the rous sarcoma virus src gene, was purified greater than 100,000-fold by a combination of ion-exchange and immunoaffinity chromatography. Incubation of pp60src purified in this fashion with [32P gamma]ATP resulted in a single 32P-labeled protein when analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Staining of these gels with silver nitrate showed a predominant 60,000-dalton polypeptide which comigrated with the protein labeled with 32P in vitro. Partial digestion of this protein with V8 protease after in vitro iodination indicated that it was pp60src. These results suggest that pp60src is able to autophosphorylate. PMID- 6283106 TI - Shope fibroma virus. II. Role of the virion-associated nucleases. AB - The effect of Shope fibroma virus (SFV) infection on host DNA synthesis was investigated. The cytocidal strain, SFV-I, inhibited the incorporation of [3H]thymidine into nuclear DNA very shortly (2 h) after infection, whereas the noncytocidal strain, SFV-W, did so later (10 h postinfection) and to a lesser extent. Furthermore, a two- to threefold stimulation of host DNA synthesis was recorded in SFV-W-infected cells 3 to 4 h after infection. Since virion associated nucleases have been implicated in the shutoff of host synthesis, these and other enzymatic activities were measured in purified virion preparations. The SFV strains and vaccinia virus contained equivalent amounts of DNA-dependent RNA polymerase, ATPase, and protein kinase activities. However, in SFV-W the pH 4.5 exonuclease activity was lower than in SFV-I and vaccinia virus, and the level of pH 7.8 endonuclease was almost undetectable. To test whether the lack of endonucleolytic activity had some effect on the removal of the cross-links in the parental DNA that occurs after viral penetration, the fate of the virion SFV DNA was followed. The majority (80%) of the SFV-I and SFV-W DNA molecules extracted after viral adsorption sedimented in alkaline sucrose gradients as cross-linked. After 3 h of infection, 75% of the SFV-I DNA molecules lacked cross-links, whereas 78% of the SFV-W DNA still remained cross-linked. The same results were obtained when the presence of cross-links was tested in restriction fragments. Taken together, these results indicate that virion-associated nucleases are involved in the early shutoff of host DNA synthesis and in the elimination of cross-links from the parental viral DNA. PMID- 6283107 TI - Glycoprotein gE of herpes simplex virus type 1: effects of anti-gE on virion infectivity and on virus-induced fc-binding receptors. AB - An Fc-binding glycoprotein, designated gE, was detected previously in cells infected with herpes simplex virus type 1 (HSV-1) and in virion preparations isolated from infected cells. For the studies reported here, we purified gE from HSV-1 strain HFEM(syn) by affinity chromatography and preparative electrophoresis and then immunized a rabbit to produce an antiserum to glycoprotein gE. We found that this antiserum selectively precipitated gE and its precursors from detergent solubilized extracts of HSV-1 strain HFEM(syn)-infected HEp-2 cells, from extracts of other cell lines infected with the same virus, and from extracts of HEp-2 cells infected with several other HSV-1 strains. The antiserum did not precipitate any proteins from uninfected cells. The several forms of gE detected by immunoprecipitation accumulated in variable quantities in different cells infected with the different virus strains and also varied slightly with respect to electrophoretic mobility, suggesting some differences in the gE's from different HSV-1 strains and some effects of the host cell on the nature and extent of post-translational processing. One of the electrophoretic forms of gE previously detected in purified preparations of virions could be precipitated by anti-gE from extracts of purified HSV-1 strain HFEM(syn) virions. Moreover, anti gE neutralized HSV-1 infectivity, but only in the presence of complement. Finally, F(ab')2 fragments of the anti-gE immunoglobulin partially inhibited the binding of 125I-labeled immunoglobulin G to the Fc receptors on HSV-1-infected cells. PMID- 6283108 TI - Similarities and differences in the Fc-binding glycoprotein (gE) of herpes simplex virus types 1 and 2 and tentative mapping of the viral gene for this glycoprotein. AB - We performed affinity chromatography and immunoprecipitation experiments to determine whether cells infected with herpes simplex virus type 2 (HSV-2) expressed a glycoprotein that was functionally and antigenically related to the HSV-1 Fc-binding glycoprotein designated gE. We found that a protein from extracts of HSV-2-infected HEp-2 cells bound specifically to an Fc affinity column and that the electrophoretic mobility of this protein in sodium dodecyl sulfate-acrylamide gels was slightly less than the mobility of HSV-1 gE. Immunoprecipitation experiments performed with an antiserum prepared against HSV 1 gE revealed that (i) extracts from HSV-2-infected cells contained a glycoprotein that was antigenically related to HSV-1 gE; (ii) the electrophoretic mobility of the HSV-2 gE was indistinguishable from the mobility of the HSV-2 Fc binding protein; (iii) the antiserum reacted with both newly synthesized transient forms and stable fully processed forms of both HSV-1 gE and HSV-2 gE; and (iv) the transient and stable forms of HSV-2 gE all had lower electrophoretic mobilities than their HSV-1 counterparts. Electrophoretic analyses of gE precipitated from extracts of HEp-2 cells infected with two sets of HSV-1 x HSV-2 intertypic recombinant viruses suggested that the gene for gE is located at the right end of the HSV genome (0.85 to 0.97 map units) in the unique portion of the S component. PMID- 6283109 TI - Characterization of N-type and dually permissive cells segregated from mouse fibroblasts whose Fv-1 phenotype could be modified by another independently segregating gene(s). AB - Though the inbred DDD mouse strain is essentially of the N type, the primary culture of this strain was about 100-fold more sensitive to B-tropic WN1802B virus than were the typical N-type strains (C3H/He, C57L, etc.). After cloning, DDD mouse cells segregated two types of cells, typical N-type cells and cells lacking in Fv-1 restriction. As both types of cells so far tested retained glucose-6-phosphatase-1 coded by a locus closely linked to Fv-1 and genetic cross experiments indicated the presence of a gene(s) modifying the Fv-1 phenotype, variation in Fv-1 restriction could presumably be brought about by genetic changes in a gene(s) other than Fv-1 itself. N-type and dually permissive cell clones were similarly established from the inbred G mouse. Compositions of polypeptides labeled with [35S]methionine in the N-type and dually permissive cells of DDD and G mouse origins were compared by two-dimensional gel electrophoresis. The polypeptide maps of these cells were similar except for a few spots. Among these dissimilar spots, a spot of about 20,000 daltons with a pI of about 5.5 was always present in N-type cells, whereas it was absent in dually permissive cells. In DDD mouse-derived clones, a proportional relation was observed between the intensity of the spot and the restriction to the B-tropic virus. PMID- 6283110 TI - Spontaneous variation and synthesis in the U3 region of the long terminal repeat of an avian retrovirus. AB - Recombinant DNA clones of a viral clone of spleen necrosis virus, an avian retrovirus, were found to have long terminal repeats of different sizes. The variation was in the U3 region of the long terminal repeats, and any one clone had U3 of the same size in both long terminal repeats. The U3 regions in the 5' and 3' long terminal repeat were shown both to be derived from the 3' long terminal repeat of parental virus DNA. PMID- 6283111 TI - In vitro and in vivo inhibition of primary transcription of vesicular stomatitis virus by a defective interfering particle. AB - A unique defective interfering (DI) particle, generated by a heat-resistant (HR) mutant of Indiana serotype vesicular stomatitis virus, was capable of inhibiting primary transcription by heterologous New Jersey serotype virions. The correlation between this phenomenon and the lowering of viral yields from doubly infected cells was investigated by the construction of chimeric DI particles containing the HR DI particle genome with a thermolabile polymerase. At the nonpermissive temperature, these DI particles were unable to self-transcribe, inhibit virion primary transcription, or reduce virion yield, but were able to be replicated. These results suggested that self-transcription of the HR DI particle genome was a prerequisite for heterotypic interference, but not for its own replication. Inhibition of virion primary transcription by HR DI ribonucleocapsids was also observed in vitro. At low HR DI to virion ribonucleocapsid ratios, the extent of inhibition was concentration dependent, whereas at high ratios, the amount of inhibition was concentration independent, approaching a limiting maximum value. A speculative mathematical model, which quantitatively accounts for these data, is presented. According to this model, the higher affinity for polymerase molecules by the HR DI ribonucleocapsids is explained in terms of dissociation events during transcription, which are more frequent in the longer virion ribonucleocapsids. PMID- 6283112 TI - Establishment of infection by spleen necrosis virus: inhibition in stationary cells and the role of secondary infection. AB - The relationship of two early events in the establishment of infection by avian retroviruses, the inhibition of viral DNA synthesis in stationary avian cells and the secondary infection which occurs after infection of replicating cells, was investigated. When neutralizing antibody to spleen necrosis virus was used to prevent secondary infection, the amount of unintegrated linear spleen necrosis virus DNA detected was much lower in infected stationary cells than in infected replicating cells. The amount of unintegrated linear spleen necrosis virus DNA in stationary cells was less than one copy per cell even at high multiplicities of infection. Viral DNA synthesis resumed after stimulation of the cells to replicate. The time of this viral DNA synthesis was closely correlated with renewed cellular DNA synthesis. In addition, blocking secondary infection of replicating cells prevented the rate of virus production from reaching the high levels usually associated with a normal productive infection by SNV. Virus production increased if secondary infection was allowed. However, this rise in virus production was not proportional to the amounts of viral DNA integrated after secondary infection. PMID- 6283113 TI - Integration site of polyoma virus DNA in the inducible LPT line of polyoma transformed rat cells. AB - The structure of the polyoma virus (Py) integration site in the inducible LPT line of Py-transformed rat cells was determined by biochemical methods of gene mapping. LPT cell DNA was digested with various restriction enzymes. The digestion products were electrophoresed in agarose gels and transferred onto nitrocellulose sheets by Southern blotting. Fragments containing viral or cell DNA sequences, or both, were identified by hybridization with Py DNA or with a cloned flanking cell DNA probe. Cleavage of LPT DNA with enzymes that restrict the Py genome once generated linear Py DNA molecules and two fragments containing both cell and viral DNA sequences. Cleavage of LPT DNA with enzymes which do not restrict Py DNA generated series of fragments whose lengths were found to differ by increments of a whole Py genome; the smallest fragment in each series was found to be longer than the viral genome. These data indicate that LPT cultures contain Py insertions of various lengths integrated into the same chromosomal site in all the cells. The length heterogeneity of the viral insertions is due to the presence of 0, 1, 2, 3. . . Py genomes arranged in a direct tandem repeat within invariable sequences of viral DNA. Double-digestion experiments were also carried out with the above enzymes and with enzymes that cleave the Py genome at multiple sites. The data obtained in these experiments were used to construct a physical map of the integration site. This map showed that the early region of the virus remained intact even in the smallest insertion (which contains no whole duplicated genomes), whereas the late region was partially duplicated and split during integration. The smallest insertion is colinear with the Py physical map over a region including the entire Py genome and at least a part of the duplicated segment. This structure could give rise to nondefective circular viral DNA molecules by single homologous recombination events. Similar recombination events may occur at a higher frequency in the longer insertions, which include longer regions of homology, and may yield many more free viral genomes. The presence of these insertions in LPT cells could thus be one of the factors which account for the high inducibility of the LPT line. PMID- 6283116 TI - Prereplicative events involving simian virus 40 DNA in permissive cells. AB - Simian virus 40 DNA molecules were found to be unable to replicate for 9 h after infection, even in cells that were already replicating the DNA of preinfecting simian virus 40; after 9 h, the ability of the DNA to replicate began to rise sharply. The kinetics of activation indicated that each DNA molecule undergoes a series of slow consecutive reactions, not involving T-antigen, before it can replicate. These pre-replicative molecular transformations probably involve configurational changes; their nature and their relation to the initiation of viral DNA synthesis is discussed. Observation of the replicative behavior of one viral DNA in the presence of another was made possible by the use of two different mutants with distinguishable DNAs: a viable deletion mutant containing DNA insensitive to TAqI restriction enzyme was used to provide viral functions required for replication, and a tsA mutant with TaqI-sensitive DNA was introduced at various times as a probe to determine the ability of the DNA to replicate under different conditions. PMID- 6283115 TI - Cloning and characterization of an envelope-specific probe from xenotropic murine leukemia proviral DNA. AB - An 8.9-kilobase EcoRI restriction fragment was cloned from mink cells chronically infected with NFS-Th-1 xenotropic murine leukemia virus by using a lambda phage host vector system. After its transfer into pBR322, the EcoRI DNA insert was characterized and found to contain 6.7 kilobases of proviral DNA sequences and 2.2 kilobases of mink cellular DNA flanking the 5' end of the viral genome. A 500 base pair fragment which was located at the 3' terminus of the cloned DNA insert and which mapped to the env region of xenotropic proviral DNA was subcloned into pBR322. This xenotropic envelope proviral DNA segment did not hybridize to ecotropic murine leukemia proviruses but did anneal to representative alpha and beta xenotropic and seven different mink cell focus-inducing proviral DNAs. The cloned xenotropic envelope-specific probe was also used in blot hybridization experiments to analyze the arrangement of related sequences in preparations of different mouse liver DNAs. PMID- 6283114 TI - Interference among defective interfering particles of vesicular stomatitis virus. AB - Three defective interfering (DI) particles of vesicular stomatitis virus (VSV), all derived from the same parental standard San Juan strain (Indiana serotype), were used in various combinations to infect cells together with the parental virus. The replication of their RNA genomes in the presence of other competing genomes was described by the hierarchical sequence: DI 0.52 particles greater than DI 0.45 particles less than or equal to DI-T particles greater than standard VSV. The advantage of one DI particle over another was not due simply to multiplicity effects nor to the irreversible occupation of limited cellular sites. Interference, however, did correlate with a change in the ratio of plus and minus RNA templates that accumulated intracellularly and with the presence of new sequences at the 3' end of the DI genomes. DI 0.52 particles contained significantly more nucleotides at the 3' end that were complementary to those at the 5' end of its RNA than did DI-T or DI 0.45 particles. The first 45 nucleotides at the 3' ends of all of the DI RNAs were identical. VSV and its DI particles can be separated into three classes, depending on their terminal RNA sequences. These sequences suggest two mechanisms, one based on the affinity of polymerase binding and the other on the affinity of N-protein binding, that may account for interference by DI particles against standard VSV and among DI particles themselves. PMID- 6283117 TI - Evidence for intramolecular self-cleavage of picornaviral replicase precursors. AB - It has previously been shown that when encephalomyocarditis viral RNA is translated in cell-free extracts of rabbit reticulocytes, it synthesizes a virus coded protease, p22, which is derived by cleavage of a precursor protein, C. Protein C is shown here to be cleaved by two different mechanisms, which were distinguished by their sensitivity to dilution. One mechanism was sensitive to dilution; the other was not. The biphasic cleavage behavior was unchanged by diluting incubation mixtures with untranslated reticulocyte extract instead of buffer, suggesting that both types of cleavage were mediated by virus translation products. It is proposed that the dilution-sensitive cleavage of protein C is due to a virus-coded protease, probably p22 itself, and that the dilution-independent cleavage is due to intramolecular self-cleavage of protein C. PMID- 6283118 TI - Avian myeloblastosis virus transforming gene is related to unique chicken DNA regions separated by at least one intervening sequence. AB - Identification of several additional restriction endonuclease sites within the cellular substitution (amv) inserted into the avian myeloblastosis virus proviral genome has permitted us to isolate different regions of the amv sequence. These subsets of the avian myeloblastosis virus transforming gene have been cloned in the plasmid pBR322 and used as hybridization probes to investigate the topology of homologous (proto-amv) normal chicken DNA sequences. The results showed that the cellular proto-amv sequences in C/O chicken DNA are interrupted by at least one intervening sequence. A partial arrangement of the proto-amv sequences is presented. PMID- 6283119 TI - Transfection of fibroblasts by cloned Abelson murine leukemia virus DNA and recovery of transmissible virus by recombination with helper virus. AB - A cloned, permuted DNA copy of the Abelson murine leukemia virus (A-MuLV) genome was capable of eliciting the morphological transformation of NIH/3T3 fibroblasts when applied to cells in a calcium phosphate precipitate. The efficiency of the process was extremely low, yielding approximately one transformant per microgram of DNA under conditions which give 10(4) transfectants per microgram of other DNAs (e.g., Moloney sarcoma virus proviral DNA). The DNA was able to induce foci, even though the 3' end of the genome was not present. The transforming gene was thus localized to the 5' portion of the genome. The transformed cells all produced viral RNA and the virus-specific P90 protein. Transmissible virus could be rescued from these cells at very low frequencies by superinfection with helper virus; the rescued A-MuLV virus had variable 3' ends apparently derived by recombination with the helper. Dimerization of the permuted A-MuLV cloned genome to reconstruct a complete provirus did not improve transformation efficiency. Virus could be rescued from these transformants, however, at a high efficiency. Cotransfection of the permuted A-MuLV DNA with proviral M-MuLV DNA yielded a significant increase in the efficiency of transformation and cotransfection of dimeric A-MuLV and proviral M-MuLV resulted in a high-efficiency transformation yielding several thousand more transformants per microgram than A-MuLV DNA alone. We propose that helper virus efficiently rescues A-MuLV from transiently transfected cells which would not otherwise have grown into foci. We hypothesize that multiple copies of A-MuLV DNA introduced into cells by transfection are toxic to cells. In support of this hypothesis, we have shown that A-MuLV DNA sequences can inhibit the stable transformation of cells by other selectable DNAs. PMID- 6283120 TI - Two large virion envelope glycoproteins mediate Epstein-Barr virus binding to receptor-positive cells. AB - The four major Epstein-Barr virion envelope components were separated by column chromatography and reconstituted into artificial liposomes. These liposomes were tested for their ability to bind selectively to Epstein-Barr virus receptor positive cells. Only when the two high-molecular-weight glycoproteins, VE1 and VE2, were present together was a stable binding complex formed. The addition of the other virion envelope components did not increase the levels of binding. This binding was inhibited by unlabeled viable virions and by neutralizing antisera, which recognized the two components. Adsorption of viable virus was also eliminated by the antisera. The enzyme susceptibility pattern of the cell liposome interaction is similar to that of the virus-cell interaction, thus confirming the specificity of the binding site. A model for Epstein-Barr virus binding in which VE1 and VE2 coordinately recognize the same binding site is presented. PMID- 6283121 TI - Comparative restriction endonuclease maps of proviral DNA of the primate type C simian sarcoma-associated virus and gibbon ape leukemia virus group. AB - Extrachromosomal DNA was purified from canine thymus cells acutely infected with different strains of infectious primate type C viruses of the woolly monkey (simian) sarcoma helper virus and gibbon ape leukemia virus group. All DNA preparations contained linear proviral molecules of 9.1 to 9.2 kilobases, at least some of which represent complete infectious proviral DNA. Cells infected with a replication-defective fibroblast-transforming sarcoma virus and its helper, a replication-competent nontransforming helper virus, also contained a 6.6- to 6.7-kilobase DNA. These proviral DNA molecules were digested with different restriction endonucleases, and the resultant fragments were oriented to the viral RNA by a combination of partial digestions, codigestion with more than one endonuclease, digestion of integrated proviral DNA, and hybridization with 3' and 5'-specific viral probes. The 3'- and 5'-specific probes each hybridized to fragments from both ends of proviral DNA, indicating that, in common with those of other retroviruses, these proviruses contain a large terminal redundancy at both ends, each of which consists of sequences derived from both the 3' and 5' regions of the viral RNA. The proviral sequences are organized 3',5'-unique 3',5'. Four restriction enzymes (KpnI, SmaI, PstI, and SstI) recognized sites within the large terminal redundancies, and these sites were conserved within all the isolates tested. This suggests that both the 3' and 5' ends of the genomic RNA of these viruses are extremely closely related. In contrast, the restriction sites within the unique portion of the provirus were not strongly conserved within this group of viruses, even though they were related along most of their genomes. Whereas the 5' 60 to 70% of the RNA of these viruses was more closely related by liquid hybridization experiments than was the 3' 30 to 40%, restriction sites within this region were not preferentially conserved, suggesting that small sequence differences or point mutations or both exist throughout the entire unique portion of the genome among these viruses. PMID- 6283122 TI - Genomic RNA of mengovirus V. Recognition of common features by ribosomes and eucaryotic initiation factor 2. AB - Binding of ribosomes to the 32P-labeled genomic RNA of mengovirus was studied in lysates of mouse L929 and Krebs ascites cells under conditions for initiation of translation. Upon total digestion with RNase T1, the 32P-labeled RNA protected in either 40S or 80S initiation complexes yielded four unique, large oligonucleotides. Each of these oligonucleotides occurred once in the viral RNA molecule. The same four oligonucleotides were recovered from 80S initiation complexes formed in lysates in which unlabeled mengovirus RNA had been translated extensively, indicating that recognition by ribosomes was not modulated detectably by a viral translation product. The recognition of intact, 32P-labeled mengovirus RNA by eucaryotic initiation factor 2 (eIF-2) was examined by direct complex formation. Fingerprint analysis of the RNA protected by eIF-2 against RNase T1 digestion yielded three T1 oligonucleotides that were identical to three of the four oligonucleotides protected in either 40S or 80S initiation complexes. A physical map of the large T1 oligonucleotides of the mengovirus RNA molecule was constructed, and the four protected oligonucleotides were found to map internally, within the region between the polycytidylate tract and the 3' end. For either ribosomes or eIF-2, the protected oligonucleotides could not be arranged in a continuous sequence, suggesting that they constitute at least two widely separated domains. These results show that ribosomes recognize and blind to more than a single sequence in mengovirus RNA, located internally in regions that are far removed from the 5' end of the molecule. eIF-2 itself binds with high specificity to mengovirus RNA, recognizing apparently three of the four sequences recognized by ribosomes. PMID- 6283123 TI - Analysis of human cytomegalovirus nucleoprotein complexes. AB - When chromatin was isolated from cells infected with human cytomegalovirus, the virus DNA remained with the chromatin fraction. If deproteinized virus DNA was added to either isolated nuclei or chromatin, the DNA was lost during the chromatin isolation. When isolated chromatin from cytomegalovirus-infected cells was banded in isopycnic metrizamide gradients, a single peak with a density of 1.18 g/cm3 was present. Analysis of this peak in isopycnic neutral CsCl gradients indicated that it contained both human cytomegalovirus and human embryonic lung cell DNAs. When infected nuclei were treated with micrococcal nuclease, 11S subunit particles which cosedimented with cell nucleosomes and contained virus DNA were isolated. PMID- 6283124 TI - Inhibition of poliovirus polymerase by guanidine in vitro. AB - Extracts from poliovirus-infected HeLa cells synthesized 35s viral RNA, replicative intermediate, and double-stranded RNA in vitro. Guanidine inhibited the synthesis of all three species of RNA; production of 35s RNA was most sensitive to the inhibitor. Pulse-chase experiments with [3H]UTP indicated that guanidine had no detectable effect on elongation of polynucleotide chains or the release of completed RNA chains from the viral replication complex. Experiments in which short pulses of precursor were used suggest that guanidine blocked the initiation step of RNA synthesis in vitro. PMID- 6283125 TI - Transformation of hamster kidney cells by fragments of BK virus DNA. AB - Hamster kidney cells were transformed, with comparable efficiency, by circular or linear molecules of complete BK virus (BKV) genome and by agarose gel-purified fragments of BKV DNA obtained by single or double digestions with various restriction endonucleases. Only fragments containing the complete early region of BKV DNA displayed transforming activity. Analysis by blot hybridization of the arrangement of viral DNA sequences in a cloned cell line transformed by a 3.8 kilobase fragment, obtained after sequential digestion of BKV DNA with HhaI and BamHI, showed the presence of seven viral integrations into the cellular DNA. Apparently all of the integrated viral molecules contained the entire early region of BKV DNA. Large T antigen, small t antigen, and the 56,000-dalton nonviral Tau antigen were detected in transformed cells by immunoprecipitation. The pattern of integration of viral sequences in transformed cells was constant over many generations. Likewise, large T antigen was always detected in transformed cells at various passage levels. These results may suggest that all of the sequences of the early region coding for large T antigen are required for transformation by BKV. Alternatively, subgenomic segments of the BKV DNA early region may be unable to transform because the appropriate polyadenylation site, necessary to obtain a complete functional transcriptional unit, is removed by the restriction enzyme cleavage. PMID- 6283126 TI - Cleavage sites within the poliovirus capsid protein precursors. AB - Partial amino-terminal sequence analysis was performed on radiolabeled polio virus capsid proteins VP1, VP2, and VP3. A computer-assisted comparison of the amino acid sequences obtained with that predicted by the nucleotide sequence of the poliovirus genome allows assignment of the amino terminus of each capsid protein to a unique position within the virus polyprotein. Sequence analysis of trypsin-digested VP4, which has a blocked amino terminus, demonstrates that VP4 is encoded at or very near to the amino terminus of the polyprotein. The gene order of the capsid proteins is VP4-VP2-VP3-VP1. Cleavage of VP0 to VP4 and VP2 is shown to occur between asparagine and serine, whereas the cleavages that separate VP2/VP3 and VP3/VP1 occur between glutamine and glycine residues. This finding supports the hypothesis that the cleavage of VP0, which occurs during virion morphogenesis, is distinct from the cleavages that separate functional regions of the polyprotein. PMID- 6283127 TI - Multimeric forms of herpes simplex virus type 2 glycoproteins. AB - Herpes simplex virus type 2-specific glycoproteins present in detergent extracts of infected cells were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis under conditions designed to permit detection of multimeric forms of these glycoproteins. Two high-molecular-weight glycosylated species were detected when samples were disrupted at lower temperatures or in the absence of any reducing agents. One multimer having an apparent molecular weight of 275,000 was identified as a multimer of the gA or gB glycoprotein or both. The second glycoprotein, having a molecular weight of approximately 230,000, was identified as a multimeric form of the gC glycoprotein. These data indicate that the gC as well as the gA and gB glycoproteins of herpes simplex virus type 2 may exist in a multimeric form. PMID- 6283128 TI - Gene protein products of SA11 simian rotavirus genome. AB - When MA104 cells were infected with SA11 rotavirus, 12 protein classes, absent in mock-infected cells, could be distinguished by polyacrylamide gel electrophoresis. At least two of these proteins were glycosylated, and their synthesis could be blocked with tunicamycin. The oligosaccharides of both glycoproteins were cleaved by endo-beta-N-acetylglucosaminidase H, suggesting that they were residues of the "high-mannose" type. Of the 12 viral polypeptides observed in infected cells, 1 was probably the apoprotein of one of these glycoproteins; 5, including 1 glycoprotein, were structural components of the virions, whereas the other 6, including a second and possibly third glycoprotein, were nonstructural viral proteins. When the 11 double-stranded RNA genome segments of SA11 were translated, after denaturation, in an RNA-dependent cell free translation system, at least 11 different polypeptides were synthesized. Ten of these polypeptides had electrophoretic migration patterns equal to those of viral proteins observed in tunicamycin-treated infected cells. Nine of the 11 double-stranded RNA genome segments were resolved by polyacrylamide gel electrophoresis and were translated individually. Two were not resolved from each other and therefore were translated together. Correlation of each synthesized polypeptide with an individual RNA segment allowed us to make a probable gene coding assignment for the different SA11 genome segments. PMID- 6283129 TI - Biochemical evidence of recombination within the unsegmented RNA genome of aphthovirus. AB - Four different pairs of temperature-sensitive mutants, derived from the same strain of aphthovirus, were crossed by using an infectious center recombination test. Each parental mutant carried an unselected marker affecting the isoelectric point of a virus-coded polypeptide; progeny of the crosses, able to grow at the nonpermissive temperature, were screened for these unselected markers. Polypeptide charge was shown to be a stable, inheritable character, and three of the four crosses yielded genetic recombinants that possessed the polypeptide marker from both parents. Peptide fingerprinting and high-resolution isoelectric focusing of the polypeptides ruled out the explantation that these viruses were generated by mutation. PMID- 6283130 TI - Mapping the in vivo arrangement of nucleosomes on simian virus 40 chromatin by the photoaddition of radioactive hydroxymethyltrimethylpsoralen. AB - Intracellular simian virus 40 (SV40) chromatin was photoreacted with a 3H-labeled psoralen derivative, hydroxymethyltrimethylpsoralen (HMT), at 48 h postinfection. Psoralen compounds have been shown to readily penetrate intact cells and, in the presence of long-wavelength UV light, form covalent adducts to DNA, preferentially at regions unprotected by nucleosomes. The average distribution pattern of [3H]HMT on the SV40 genome was determined by specific activity measurements of the DNA fragments generated by HindIII plus HpaII or by AtuI restriction enzyme digestion. At levels of 1 to 10 [3H]HMT photoadducts per SV40 molecule, the radiolabel was found to be distributed nonrandomly. Comparison of the labeling pattern in vivo with that of purified SV40 DNA labeled in vitro revealed one major difference. A region of approximately 400 base pairs, located between 0.65 and 0.73 on the physical map, was preferentially labeled under in vivo conditions. This finding strongly suggests that the highly accessible region near the origin of replication, previously observed on isolated SV40 "minichromosomes," exists on SV40 chromatin in vivo during a lytic infection. PMID- 6283131 TI - Shope fibroma virus. I. Biological and molecular properties of a cytocidal and a noncytocidal strain. AB - The biological and molecular properties of two strains of Shope fibroma virus (SFV) were compared. SFV-I was highly cytocidal to most of the cell lines tested and produced pocks in the chorioallantoic membrane of chick embryos. By contrast, SFV-W did not produce cytopathic effects in any of the cell lines or in the chorioallantoic membrane, but it induced characteristic foci 3 to 4 days after infection. Both strains produced tumors when inoculated into the skin of susceptible rabbits. Maximal infectivity in BSC-1 cells was reached by both strains between 24 to 48 h after inoculation. Viral DNA synthesis also took place at the same time, although cells infected with SFV-I incorporated three times more [(3)H]thymidine than cells infected with SFV-W. Sedimentation analysis and hydroxylapatite chromatography of the two viral DNAs indicated that their molecular weights were similar and that both were naturally cross-linked. Digestion with three restriction endonucleases, however, revealed that they had different restriction sites. When SFV-I and vaccinia DNA were compared, the restriction patterns were more alike. Analysis of the virion structural proteins by gel electrophoresis indicated that SFV-I, SFV-W, and vaccinia virus had many polypeptides in common, although there were distinctive differences among the three viruses. Finally, the results of plaque neutralization tests with different antisera showed that SFV-I and SFV-W shared common antigens and that vaccinia antiserum inhibited SFV-I but not SFV-W. We conclude that the SFV-I genome contains information for both cytolysis and tumorigenesis. This unusual virus may be a recombinant between an orthopoxvirus and a leporipoxvirus. PMID- 6283132 TI - Candidate product of the FBJ murine osteosarcoma virus oncogene: characterization of a 55,000-dalton phosphoprotein. AB - Sera from rat bearing tumors induced by inoculation of FBJ murine osteogenic sarcoma virus (FBJ-MSV) nonproducer rat cells precipitate two proteins with molecular weights of 55,000 (p55) and 39,000 (p39) from FBJ-MSV-transformed cells. These proteins cannot be precipitated from uninfected cells or cells transformed by other strains of murine sarcoma virus, nor can they be precipitated by sera specific for the viral structural proteins. A methionine tryptic peptide mapping analysis showed that p55 and p39 have little or no homology and that they are not related to the helper virus gag and env gene products. p55 could also be detected among the in vitro translation products of 70S RNA from FBJ murine leukemia virus plus FBJ-MSV virions but not among those from FBJ murine leukemia virus alone. This suggests that p55 is encoded by the FBJ-MSV genome, whereas p39, which was not detected among the in vitro translation products, may not be virus encoded. Another difference between p55 and p39 is that p55 is phosphorylated, with most of the phosphate on a serine residue(s), whereas p39 is phosphorylated to a much lesser extent, if at all. No protein kinase activity was associated with p55 and p39 immune complexes under standard conditions. Our data suggest that p55 is a strong candidate for the FBJ MSV oncogene product. PMID- 6283133 TI - Mechanism of replication of bacteriophage phi X174 XX. Sensitivity of nascent DNA to single-strand-specific nucleases. AB - We reported earlier that dephosphorylated nascent phi X174 viral strand DNA molecules were less extensively degraded from the 5' end by spleen exonuclease than were non-nascent molecules. Experiments described here revealed that the insensitivity to the 5'-OH end-specific nuclease was more evident among the longer molecules in the population than among the shorter, all of the molecules being less than unit length in size. The smallest molecules in the population were about as sensitive to the enzyme as the control molecules and hence must possess unblocked 5'-terminal nucleotides. Degradation of the nascent DNA with the 3' end-specific snake venom phosphodiesterase revealed only a small enrichment for [3H]thymidine near the 3' end, seemingly insufficient to account completely for the apparent insensitivity of the longer molecules to spleen exonuclease. When the nascent molecules were isolated without the use of proteolytic enzymes, some pronase-sensitive material was found associated with the DNA, particularly the longer molecules. We suggest that the resistance of the longer nascent (pronase-treated) molecules to spleen exonuclease occurs because they have remnants of the viral gene A or A* protein covalently bound to the 5' end. PMID- 6283134 TI - Intracisternal A-particle genes: structure of adjacent genes and mapping of the boundaries of the transcriptional unit. AB - Adjacent intracisternal A-particle (IAP) genes were identified in two different recombinant DNA clones, gamma 81 and gamma 19. In clone gamma 81, the most common form of IAP gene was separated by 5.3 kilobases from another IAP gene that had two apparent internal deletions. The two genes were in a head-to-tail configuration. In clone gamma 19, two different types of IAP genes were separated by less than 0.5 kilobase. Blot hybridization analysis of mouse DNA demonstrated that the DNA sequence found in clone gamma 81 is identical to the in vivo configuration. Using isolated DNA fragments from clone gamma 19, we mapped the boundaries of the IAP RNA by S1 digestion of RNA-DNA hybrids and by cDNA extension. With these techniques, both the 5' end and the 3' end of the IAP RNA in two different plasmacytomas (MOPC 315 and TEPC 15) were shown to fall within the long terminal direct repeat of the IAP gene. The fragment sizes generated by S1 digestion of IAP RNAs isolated from the two tumor lines were found to differ, indicating that different IAP genes may be transcribed in these two plasmacytomas. PMID- 6283135 TI - RNA-dependent RNA polymerase activity in coronavirus- infected cells. AB - An enzymatic activity which incorporates [3H]UMP into acid-precipitable material in the presence of endogenous template was found in the cytoplasm of porcine cells infected with the transmissible gastroenteritis virus of swine. This activity was not found in uninfected control cells, nor was it found in purified virus. The activity was associated with the mitochondrial fraction of infected cells, suggesting that the enzyme is membrane bound. The activity required the presence of all three ribonucleoside triphosphates in addition to [3H]UTP, and it was not inhibited by actinomycin D. The heated product was digested by RNase but not by DNase. Mg2+ was required for enzymatic activity, and its optimal concentration was approximately 5 mM. The size of the in vitro products was compared by electrophoresis with that of in vivo-synthesized virus-specified RNA to confirm the viral specificity of the polymerase activity. Virus-specified RNA from infected cells consisted of 10 species of single-stranded, polyadenylated RNA with molecular weights of 6.8 X 10(6), 6.2 X 10(6), 3.15 X 10(6), 1.40 X 10(6), 1.05 X 10(6), 0.94 X 10(6), 0.66 X 10(6), 0.39 X 10(6), 0.34 X 10(6), and 0.24 X 10(6). In vitro synthesized RNA consisted of a high-molecular-weight species, of apparently higher molecular weight than genomic RNA, and two single stranded species that electrophoretically comigrated with the species of 1.40 X 10(6) and 0.66 X 10(6) molecular weight made in vivo. PMID- 6283136 TI - Continuing germ line integration of AKV proviruses during the breeding of AKR mice and derivative recombinant inbred strains. AB - The gel electrophoresis-hybridization technique of Southern was used to analyze genetically transmitted proviruses coding for the AKV strain of murine leukemia virus. We were able to identify the restriction endonuclease EcoRI fragments containing two previously unidentified, genetically transmitted AKV proviruses of AKR mice. Comparison of different sublines of AKR mice revealed considerable heterogeneity in their complement of germ line proviruses. This heterogeneity provides evidence that the provirus complement of AKR mice is not stable. Rather, the number of genetically transmitted proviruses increases during inbreeding. Examination of a series of sublines of the C3H strain indicated that this amplification is dependent on viremia. We estimate that, in viremic strains of mice, one new provirus becomes fixed in the germ line every 15 to 30 years. PMID- 6283137 TI - Influenza B virus genome: sequences and structural organization of RNA segment 8 and the mRNAs coding for the NS1 and NS2 proteins. AB - Double-stranded DNA derived from influenza B virus genome RNA segment 8, which codes for the NS1 and NS2 proteins, was constructed by hybridization of full length cDNA copies of RNA segment 8 and of the NS1 mRNA. This DNA was cloned in plasmid pBR322 and sequenced. The NS1 mRNA (approximately 1,080 viral nucleotides) contains nonviral nucleotides at its 5' end and is capable of coding for a protein of 281 amino acids. Sequencing of the NS2 mRNA has shown that it contains an interrupted sequence of 655 nucleotides and is most likely synthesized by a splicing mechanism. The first approximately 75 virus-specific nucleotides at the 5' end of the NS2 mRNA are the same as are found at the 5' end of the NS1 mRNA. This region contains the initiation codon for protein synthesis and coding information for 10 amino acids common to the two proteins. The approximately 350-nucleotide body region of the NS2 mRNA can be translated in the +1 reading frame, and the sequence indicates that the NS1 and NS2 protein coding regions overlap by 52 amino acids translated from different reading frames. Thus, between the influenza A and B viruses, the organization of the NS1 and NS2 mRNAs and the sizes of the NS2 mRNA and protein are conserved despite the larger size of the influenza B virus RNA segment, NS1 mRNA, and NS1 protein. PMID- 6283138 TI - Carboxy-terminal analysis of poliovirus proteins: termination of poliovirus RNA translation and location of unique poliovirus polyprotein cleavage sites. AB - The carboxy-terminal amino acids of a number of poliovirus proteins were determined by carboxypeptidase A analysis. The nonstructural proteins P3-2, P3-4b and their precursor. P3-1b, were found to be coterminal with a sequence of -Ser Phe-COOH. As these proteins are coded for at the extreme 3' end of the viral RNA, it is possible to establish the termination site of translation at nucleotide 7,361, 73 nucleotides before the start of the polyadenylic acid tract of the RNA. Two additional nonstructural proteins, P2-X and its precursor, P2-3b, were also found to be coterminal with a sequence of -Phe-Gln-COOH. This result confirms the existence of at least one Gln-Gly proteolytic cleavage site. These Gln-Gly cleavage sites are predicted from the nucleotide sequence to be ubiquitous throughout the poliovirus genome. The only exceptions are the cleavage sites at the carboxy termini of the structural protein VP4 and VP1. Carboxypeptidase A analysis of VP1 establishes a terminal sequence of -Thr-Tyr-COOH, and similar analysis of VP4 shows Asn to be the terminal amino acid residue, observations that prove the existence of the exceptional C-terminal amino acids. In none of the analyzed cases has C-terminal trimming after cleavage been observed. PMID- 6283139 TI - Isolation and preliminary characterization of a phosphonoacetic acid-resistant and temperature-sensitive mutant of herpes simplex virus type 1. AB - A group of 43 phosphonoacetic acid (PAA)-resistant mutants of herpes simplex virus type 1 was isolated after the mutagenesis of infected cells with nitrosoguanidine. One of these mutants, designated PAA1rts1, was found to be temperature sensitive (ts), that is, unable to replicate at 39.5 degrees C, the nonpermissive temperature. Recombination analysis of PAA1rts1 indicated that the PAA1r mutation and the ts1 mutation are loosely linked and are located on two separate genes. PAA1rts1 showed a defect in viral DNA synthesis at 39.5 degrees C, which presumably can be attributed to the production of a PAA-resistant and thermolabile DNA polymerase. PAA1rts1 was also defective in the shutoff of host DNA synthesis at the restrictive temperature. PMID- 6283140 TI - Association of cap-binding protein with eucaryotic initiation factor 3 in initiation factor preparations from uninfected and poliovirus-infected HeLa cells. AB - Extracts from poliovirus-infected HeLa cells are unable to translate vesicular stomatitis virus or cellular mRNAs in vitro, probably reflecting the poliovirus induced inhibition of host cell protein synthesis which occurs in vivo. Crude initiation factors from uninfected HeLa cells are able to restore translation of vesicular stomatitis virus mRNA in infected cell lysates. This restoring activity separates into the 0 to 40% ammonium sulfate fractional precipitate of ribosomal salt wash. Restoring activity is completely lacking in the analogous fractions prepared from poliovirus-infected cells. The 0 to 40% ammonium sulfate precipitates from both uninfected and infected cells contain eucaryotic initiation factor 3 (eIF-3), eIf-4B, and the cap-binding protein (CBP), which is detected by means of a cross-linking assay, as well as other proteins. The association of eIF-3 and cap binding protein was examined. The 0 to 40% ammonium sulfate precipitate of ribosomal salt wash from uninfected and infected cells was sedimented in sucrose gradients. Each fraction was examined for the presence of eIF-3 antigens by an antibody blot technique and for the presence of the CBP by cross-linking to cap-labeled mRNAs. From uninfected cells, a major proportion of the CBP cosedimented with eIF-3; however, none of the CBP from infected cells sedimented with eIF-3. The results suggest that the association of the CBP with eIF-3 into a functional complex may have been disrupted during the course of poliovirus infection. PMID- 6283141 TI - Structural analysis of virion proteins of the avian coronavirus infectious bronchitis virus. AB - We have found six major polypeptides in virions of the avian coronavirus infectious bronchitis virus grown in tissue culture: four glycoproteins, GP84, GP36, GP31, and GP28, and two non-glycosylated proteins, P51 and P23. In addition, we detected three minor species: two glycoproteins, GP90 and GP59, and one non-glycosylated protein, P14. Two-dimensional tryptic peptide mapping showed that GP36, GP31, GP28, and P23 comprise a group of closely related proteins which we have designated the "P23 family," but that the other proteins are distinct. Analysis by partial proteolytic digestion of P23 family, but that the other proteins are distinct. Analysis by partial proteolytic digestion of the P23 family labeled biosynthetically with [35S] methionine, and P23, labeled with [35S] formyl-methionine by in vitro translation of RNA from infected cells, revealed that the proteins of the P23 family differ in their amino-terminal domains. Similar analysis of GP31 and Gp36 labeled with [3H] mannose showed that the partial proteolytic fragments unique to these proteins were glycosylated. This suggests that differences in glycosylation in the amino-terminal domains contributes to the marked polymorphism os the P23 family. The results are discussed with respect to possible models for synthesis of the virion proteins. PMID- 6283143 TI - Identification of a precursor in the biosynthesis of the p21 transforming protein of harvey murine sarcoma virus. AB - The p21 transforming protein coded for by the v-ras gene of Harvey murine sarcoma virus (Ha-MuSV) migrates as a doublet band between 21,000 and 23,000 daltons during sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The lower band of the doublet is designated p21, and the upper band is designated pp21 since it comigrates with the phosphorylated form of p21. By pulse-labeling with [35S] methionine, we detected a p21 precursor, pro-p21, which migrated as if it was approximately 1,000 daltons larger than p21. The precursor-product relationship was established by pulse-chase experiments with [25S] methionine in the presence of 100 micrograms of cycloheximide per ml, which inhibited all de novo protein biosynthesis. Within 4 h, pro-p21 was completely chased into p21, and during the next 24 h pp21 accumulated. Thus, formation of pp21 from p21 did not require de novo protein synthesis. By subcellular fractionation into cytosol amd membrane fractions, we found that pro-p21 was synthesized in a non-membrane-bound state and that shortly after its complete synthesis, the p21 product was associated with the membrane fraction. By selective cleavage of p21 at a unique aspartic acid-proline residue with 70% formic acid or with Staphylococcus aureus V8 protease, we found that the intramolecular site of pro-p21 processing was located in the C-terminal portion of the pro-p21 molecule. The possibilities that the precursor was involved in the assembly of p21 into the plasma membrane and, alternatively, that the processing was a step in the activation of p21 biochemical activities are discussed. PMID- 6283142 TI - Genome of reticuloendotheliosis virus: characterization by use of cloned proviral DNA. AB - Reticuloendotheliosis virus is an avian type C retrovirus that is capable of transforming fibroblasts and hematopoietic cells both in vivo and in vitro. This virus is highly related to the three other members of the reticuloendotheliosis virus group, including spleen necrosis virus, but it is apparently unrelated to the avian leukosis-sarcoma virus family. Previous studies have shown that it consists of a replication-competent helper virus (designated REV-A) and a defective component (designated REV) that is responsible for transformation. In this study we used restriction endonuclease mapping and heteroduplex analysis to characterize the proviral DNAs of REV-A and REV. Both producer and nonproducer transformed chicken spleen cells were used as sources of REV proviral DNA; this genome was mapped in detail, and fragments of it were cloned in lambdagtWES.lambdaB. The infected canine thymus line Cf2Th(REV-A) was used as a source of REV-A proviral DNA. The restriction maps and heteroduplexes of the REV and REV-A genomes showed that (proceeding from 5' to 3') (i) REV contains a large fraction of the REV-A gag gene (assuming a gene order of gag-pol-env and gene sizes similar to those of other type C viruses), for the two genomes are very similar over a distance of 2.1 kilobases beginning at their 5' termini; (ii) most or all of REV-A pol is deleted in REV; (iii) REV contains a 1.1 kilobase segment derived from the 3' end of REV-A pol or the 5' end of env or both; (iv) this env region in REV is followed by a 1.9-kilobase segment which is unrelated to REV-A; and (v) the helper-unrelated segment of REV extends essentially all of the way to the beginning of the 3' long terminal repeat. Therefore, like avian myeloblastosis virus but unlike the other avian acute leukemia viruses and most mammalian and avian sarcoma viruses, REV appears to be an env gene recombinant. We also found that the REV-specific segment is derived from avian DNA, for a cloned REV fragment was able to hybridize with the DNA from an uninfected chicken. Therefore, like the other acute transforming viruses, REV appears to be the product of recombination between a replication-competent virus and host DNA. Two other defective genomes in virus-producing chicken cells were also cloned and characterized. One was very similar to REV in its presumptive gag and env segments, but instead of a host-derived insertion it contained additional env sequences. The second was similar (but not identical) to the first in its gag and env regions and appeared to contain an additional 1-kilobase inversion of REV-A sequences. PMID- 6283144 TI - Simian virus 40-rabbit beta-globin recombinants lacking late mRNA splice sites express cytoplasmic RNAs with altered structures. AB - Deletions were introduced at exon-intron boundaries in the late region of a simian virus 40-beta-globin cDNA recombinant to study the role of splicing in the formation of simian virus 40 late cytoplasmic RNAs. The recombinant was used as a wild type because it allowed characterization of mutant RNAs expressed from defective genomes in the presence of comparable RNAs contributed by the coinfecting helper virus. Removal of a 17-base pair segment at map position 0.76, which included a portion of the leader sequence implicated in the splicing of the major 16S mRNA, prevented expression of 16S-type mRNA. The same mutant accumulated cytoplasmic 19S-type mRNA, but the assortment of the 5' ends of these mRNAs differed from the assortment of the wild-type counterparts. Another mutant that lacks nucleotide sequences implicated in the splicing of the major 16S mRNA and one of the principal 19S-type RNAs accumulated a 16S-type mRNA with a previously undetected leader splice, and assortment of 19S mRNAs with new or normally underrepresented splices, and even a species of unspliced cytoplasmic 19S mRNA. PMID- 6283145 TI - Detection of herpes simplex virus type 2 glycoproteins expressed in virus transformed rat cells. AB - Rat embryo fibroblasts transformed by herpes simplex virus type 2 (HSV-2) were assayed for the expression of certain virus-specific glycoproteins on the surface membranes. Monospecific antisera to HSV-2-specific glycoproteins, designated gAgB, gC, and gX, were used in membrane immunofluorescence studies with HSV-2 transformed cell lines tREF-G-1, tREF-G-2, and a tumor-derived rat fibrosarcoma cells line produced in syngeneic rats inoculated with tREF-G-1 cells. Analysis of the three HSV-2-transformed cell lines showed that antisera to the gAgB and gX glycoproteins were reactive with these cells. In contrast, no significant reactivity was observed when anti-gC serum was reacted with the HSV-2-transformed cell lines. All three antiglycoprotein sera reacted positively with rat cells productively infected with HSV-2. Additionally, the HSV-2-transformed and tumor derived cell lines showed positive internal immunofluorescence after reaction with antiserum to an early, nonstructural viral protein designated VP143 (molecular weight, 143,000). Infectivity of HSV-2 in standard plaque assays was neutralized by hyperimmune rat antisera to tREF-G-2 or rat fibrosarcoma cells and to HSV-2 virions and by sera from rats bearing the fibrosarcoma. Adsorption of rat-anti-HSV-2 serum with tREF-G-2 or rat fibrosarcoma cells reduced neutralizing activity to 10 and 12%, respectively, compared with 90% neutralization by antiserum adsorbed with nontransformed rat embryo fibroblast cells and 100% neutralization with unadsorbed antiserum. In summary, HSV-2-transformed rat cells retained and expressed genetic information necessary for the production of HSV-2 glycoproteins and a nonstructural protein after high passage in tissue culture or in the syngeneic host. PMID- 6283146 TI - New way to isolate simian virus 40 nucleoprotein complexes from infected cells: use of a thiol-specific reagent. AB - A new method for the isolation of simian virus 40 nucleoprotein complexes from nuclei of lytically infected cells is described. The method is based on the addition of a thiol-specific reagent, 5'5'-dithiobis(2-nitrobenzoic acid), to lysis and extraction buffers. By inhibiting an uncoating activity during simian virus 40 extraction, 5'5'-dithiobis (2-nitrobenzoic acid) allows the use of efficient extraction buffers, such as one containing Triton X-100 and EDTA, for the isolation of native simian virus 40 minichromosomes and virion-type structures. Use of the method is illustrated by following encapsidation of simian virus 40 minichromosomes in a pulse-chase experiment. Since 5'5'-dithiobis (2 nitrobenzoic acid) is an inhibitor of many different enzymes, the 5',5'-dithiobis (2-nitrobenzoic acid) extraction technique may be useful for the isolation of not only papovaviruses but also other viruses and possibly cellular chromatin. PMID- 6283147 TI - Electron microscopic study of equine herpesvirus type 1 DNA. AB - Electron microscopic studies of equine herpesvirus DNA revealed that single strands that were allowed to reanneal formed single-stranded loops with double stranded stems only at one end of the molecule. These observations support restriction enzyme analyses which indicate that the 92-megadalton DNA molecule exists as a long region of unique sequences covalently linked to a short region. The short region is comprised of an internal unique sequence, which forms the loop during reannealing of single strands, and two terminal inverted repeat sequences that bracket the unique sequence and form the double-stranded stem structure observed upon reannealing of single strands. Measurements of the unique sequence and terminal inverted repeat subgenomic sequences indicate a size of 6.4 megadaltons for each and thus fix the size of the short region at approximately 19.2 megadaltons. PMID- 6283149 TI - Linear mapping of tryptophan residues in Vesiculovirus M and N proteins by partial chemical cleavage. AB - Nonlimit chemical cleavage at tryptophan residues of protein labeled at the amino terminus afforded a simple procedure for generating specific fragments and for mapping tryptophan positions. A comparison of the matrix (M) and nucleocapsid (N) proteins of four members of the Vesiculovirus group by this procedure suggests considerable conservation of tryptophan number and location in the four serotypes examined. PMID- 6283148 TI - Sequence homology between avian and human adenoviruses. AB - Studies of hybridization between fowl adenovirus type 1 (chicken embryo lethal orphan virus) DNA and human adenovirus type 2 DNA revealed two short but distinct regions which cross-hybridized under stringent conditions. One of the homologous regions was located between map positions 18.1 and 19.3 and did not correspond to any gene recognized so far. The second region mapped in the hexon gene between position 57 and 58. PMID- 6283150 TI - Integrated adenovirus type 12 DNA in the transformed hamster cell line T637: sequence arrangements at the termini of viral DNA and mode of amplification. AB - Approximately 20 to 22 copies of adenovirus type 12 (Ad12) DNA per cell were integrated into the genome of the cell line T637. Only a few of these copies seemed to remain intact and colinear with virion DNA. All other persisting viral genomes exhibited deletions or inversions or both in the right-hand part of Ad12 DNA. Spontaneously arising morphological revertants of T637 cells has lost viral DNA. In most of the revertant cell lines only the intact or a part of the intact viral genome was preserved; other revertant cell lines has lost all viral DNA. In three other Ad12-transformed hamster cell lines, HA12/7, A2497-3, and CLAC3 (Stabel et al., J. Virol. 36:22-40, 1980), major rearrangements at the right end of the integrated Ad12 DNA were not found. These studies were performed to investigate the phenomena of amplification, rearrangements, and deletions of Ad12 DNA in hamster cells. PMID- 6283151 TI - Shedding of vesicular stomatitis virus soluble glycoprotein by removal of carboxy terminal peptide. AB - A comparison of partial NH2-terminal sequences of vesicular stomatitis viral glycoprotein G (molecular weight, 69,000) and the soluble extracellular glycoprotein antigen Gs (molecular weight, 57,000) shows that both of the sequences are identical. Tryptic fingerprint analyses show that Gs lacks the carboxy-terminal region containing the membrane-anchoring hydrophobic domain of G. These results suggest that Gs is formed by cleavage in the carboxy-terminal region of G. PMID- 6283152 TI - Synthesis of murine leukemia viral DNA in vitro: evidence for plus-strand DNA synthesis at both ends of the genome. AB - We studied the synthesis of B-tropic murine leukemia viral DNA in vitro by detergent-disrupted virions. The reaction products (detected by the Southern transfer technique) included full-length, infectious, double-stranded DNA and several subgenomic fragments. Restriction endonuclease analysis and hybridization and specific probes revealed two classes of subgenomic fragments: some were derived from the right end of the genome, and some were derived from the left end. Most of the fragments harbored one long terminal repeat copy at their ends, suggesting that they were initiated correctly. S1 nuclease and restriction endonuclease treatments of these fragments indicated that a single-stranded gap was present near the first initiation site of plus strong-stop DNA. The treatments also suggested the presence of a second initiation site flanked by a single-stranded gap 0.9 kilobase pairs from the right end of the genome. Our data clearly show that plus-strand DNA is synthesized at both ends of the genome, by using plus strong stop as the first initiation site and additional initiation sites. PMID- 6283153 TI - Fv-1 determinants in xenotropic murine leukemia viruses studied with biological assay systems: Isolation of xenotropic virus with N-tropic Fv-1 activity in the cryptic form. AB - By a biological assay system using phenotypically mixed ecotropic and xenotropic murine leukemia viruses, we investigated whether in the virions of a xenotropic virus there is N- or B-tropic Fv-1 determinant in active form. The existence of N tropic Fv-1 determinant was demonstrated in SL-XT-1 xenotropic virus isolated from the spleen of a 3-month-old SL mouse, and the N-tropic Fv-1 tropism was confirmed by analysis of the phenotypically mixed viruses harvested from clonal SC-1 cells doubly infected with the SL-XT-1 and B-tropic ecotropic viruses. However, neither N- nor B-tropic Fv-1 determinant was demonstrated in any xenotropic viruses isolated from embryo cells of BALB/c, NZB, or DBA/2 mice, or Cas E #1-IU, and xenotropic-like virus isolated from a wild mouse. PMID- 6283154 TI - Structure of a replication intermediate in the synthesis of Rous sarcoma virus DNA in vivo. AB - Intermediates in the synthesis of Rous sarcoma virus DNA in vivo contain a short second strand of DNA (plus strong-stop DNA) synthesized by using the region near the 5' end of the first (minus) strand of DNA as the template. In this report, we show that the 3' end of plus strong-stop DNA is extended about 15 to 20 nucleotides beyond the 5' end of the minus-strand DNA template, probably copying a portion of the tRNATrp molecule that serves as primer for synthesis of the minus strand of DNA. The extra sequences present in plus strong-stop DNA may play a central role in the generation of the long terminal repeat present in mature forms of viral DNA. PMID- 6283155 TI - NS phosphoprotein of vesicular stomatitis virus: subspecies separated by electrophoresis and isoelectric focusing. AB - The NS protein of vesicular stomatitis virus is the only phosphorylated nucleocapsid protein. The amount of NS phosphorylation appears to regulate the activity of the protein in the transcription of the virus genome. Several methods have been used to separate NS subspecies containing different amounts of phosphate, but the relationships among the subspecies separated by different workers have been unclear. We report that the isoelectric points of NS molecules were abnormally acidic in some commercial ampholytes, but favorable ampholytes resolved multiple phosphorylated NS subspecies with isoelectric points ranging from pH 6.8 to 7.2. The most highly phosphorylated NS molecules had more acidic isoelectric points, and they exhibited greater electrophoretic mobilities in two previously employed electrophoretic systems. PMID- 6283157 TI - Subgenomic mRNA in OK10 defective leukemia virus-transformed cells. AB - OK10, a defective leukemia virus, is produced as a defective particle by so called nonproducer transformed quail fibroblasts. OK10 defective viral particles contain an 8-kilobases (kb)-long genomic RNA, lack any detectable reverse transcriptase activity, and are not infectious. We studied the genetic content of OK10 RNA extracted from both virions and infected cells. As shown by RNA-cDNA hybridizations in stringent conditions, about 77% (6.4 kb) of the OK10 8.0kb RNA was related to avian leukosis viruses in the three structural genes gag, pol, and env, as well as in the c region. The remainder of the OK10 genome-encoding capacity ( AMP --> IMP --> inosine --> hypoxanthine; conversion of IMP to AMP and de novo purine synthesis were inhibited in these cells. ATP degradation did not occur in a mutant of CEM that was incapable of phosphorylating deoxyadenosine, or in a B cell line with very limited ability to accumulate dATP. We found that dATP and ATP were both able to stimulate markedly the deamination of AMP by lymphoblast AMP deaminase; dAMP was a poor substrate for this enzyme (K(m) = 2.4 mM, vs. 0.4 mM for AMP). Similarly, dATP as well as ATP caused marked activation of IMP dephosphorylation by a lymphoblast cytoplasmic nucleotidase. Inhibition of intracellular AMP deaminase with coformycin prevented degradation of adenine ribonucleotides without affecting dATP accumulation. We propose that ATP-dependent phosphorylation of deoxyadenosine generates ADP and AMP. Simultaneously, dATP accumulation stimulates deamination of AMP, but not dAMP, and the dephosphorylation of IMP to inosine. Coupling of AMP degradation to ATP utilization in deoxyadenosine phosphorylation maintains the adenylate energy charge despite net depletion of cellular ATP. PMID- 6283542 TI - DNA-mediated gene transfer: recombination between cotransferred DNA sequences and recovery of recombinants in a plasmid. AB - Ltk- aprt- mouse L cells were transformed to the tk+ phenotype with 10 ng of the herpes simplex virus-1 thymidine kinase (tk) gene and 20 micrograms of pBR322 or simian virus 40 (SV40) DNA. DNAs from five cloned cell lines show restriction endonuclease fragments that hybridize to both tk and pBR322 or SV40 DNA. In all of the cell lines some of these fragments also contain cellular DNA sequences. The use of carrier DNAs with defined sequences has enabled us to demonstrate that the joining of carrier and selectable gene sequences occurs in mouse cells. In one case we have been able to use the ampicillin resistance marker of pBR322 to "rescue" a recombinant plasmid. An analysis of the junction between pBR322 and tk in this plasmid suggests that a small area of homology (16 of 19 base pairs) might be involved in the recombination process. PMID- 6283541 TI - Site of premature termination of late transcription of simian virus 40 DNA: enhancement by 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole. AB - Sedimentation analysis of pulse-labeled RNA synthesized in nuclei isolated from simian virus 40-infected cells revealed an abundance of short cellular and viral RNAs. The relative amount of the short chains is increased in nuclei isolated from cells treated with 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole (DRB). The short viral RNAs were purified by hybridization to and elution from simian virus 40 DNA on filters, and their sizes were determined by gel electrophoresis. A major band of 93- to 95-nucleotide-long RNA was observed along with additional minor bands. Identical bands were revealed when the viral RNA was purified from nuclei of cells pretreated with DRB. The major band was identified as an aborted transcript of a RNA that initiated at the major initiation site (nucleotide 243). We have found that the DNA region where the RNA stops is A+T rich and is immediately preceded by a G+C-rich region that exhibits dyad symmetry, resembling the termination signal in prokaryotes. These observations show that RNA polymerase II responds to the same termination signal as the prokaryotic enzyme and suggest that a mechanism of attenuation regulates simian virus 40 late transcription. PMID- 6283543 TI - The beta-adrenergic receptor: rapid purification and covalent labeling by photoaffinity crosslinking. AB - New procedures for the rapid purification and covalent labeling of the beta adrenergic receptors have been developed that should greatly accelerate progress in the study of these widely distributed adenylate cyclase-coupled receptors. Chromatography of solubilized receptor preparations on a Sepharose-alprenolol affinity gel followed by HPLC on steric exclusion columns lead to rapid (2 days) and high yield (approximately 30%) purification of the receptors from frog erythrocytes. The receptor obtained by these rapid procedures appears to be composed entirely of 58,000 Mr subunit(s) and to be identical to that previously purified by much lengthier procedures [Shorr, R. G. L., Lefkowitz, R. J. & Caron, M. G. (1981) J. Biol. Chem. 256, 5820-5826]. A novel, very high affinity, specific beta-adrenergic antagonist, p-aminobenzylcarazolol, has also been synthesized. It can be radioiodinated to theoretical specific radioactivity with 125I (2,200 Ci/mmol). This radioligand, which possesses an arylamine moiety, may then be covalently incorporated into the receptor binding subunit (58,000 Mr peptide) of the frog erythrocyte membranes by the use of the bifunctional photoactive crosslinker N-succinimidyl-6-(4'-azido-2'- nitrophenylamino)hexanoate (SANAH). Covalent incorporation is blocked by various drugs with a strict beta adrenergic specificity. This suggests that the photoaffinity crosslinking approach may be useful for labeling a variety of small molecule and neurotransmitter receptors when appropriate ligands can be synthesized. PMID- 6283544 TI - Purification of two calcium/calmodulin-dependent forms of cyclic nucleotide phosphodiesterase by using conformation-specific monoclonal antibody chromatography. AB - A procedure for nondenaturing immunopurification of bovine calmodulin-dependent 3',5'-cyclic-nucleotide phosphodiesterase (3',5'-cyclic-nucleotide 5' nucleotidohydrolase, EC 3.1.4.17) is described that utilizes chromatography on a conformation-specific monoclonal antibody column. Hybridomas derived from spleen cells of mice immunized with Ca(2+)/calmodulin/phosphodiesterase were screened for antiphosphodiesterase antibody production. A stable cell line was established that secretes a monoclonal antibody that binds to the Ca(2+)/calmodulin/enzyme complex with an approximate K(d) of 10(-9) M. The dissociation constant was increased by two orders of magnitude when calmodulin interaction with the enzyme was inhibited by Ca(2+) chelation. This differential reactivity was utilized for affinity chromatography of heart and brain phosphodiesterases on monoclonal antibody columns. Highly purified phosphodiesterases were eluted in good yield with buffer containing EGTA. The immunopurified enzymes from heart and brain exhibited specific activities of approximately 300 units/mg when assayed at millimolar concentrations of cGMP or cAMP. Calmodulin stimulated both enzymes 10- to 15-fold over basal activity under these conditions. However, analysis of the two preparations by NaDodSO(4)/polyacrylamide gel electrophoresis revealed an apparent subunit of M(r) 61,000 for the brain enzyme, in contrast to the M(r) 59,000 cardiac subunit. The observed difference was not an artifact of tissue homogenization because both forms were detected after purification from mixed tissue homogenates. These results suggest that mild, biospecific elution from a conformation-specific monoclonal antibody column may be a general technique applicable to the rapid isolation of proteins whose antigenic determinants can be altered with specific ligands. PMID- 6283545 TI - ATP-driven sodium pump in Streptococcus faecalis. AB - Sodium extrusion by bacteria is generally attributed to secondary antiport of Na+ for H+ energized by the proton circulation. Streptococcus faecalis is an exception, in that sodium expulsion from intact cells requires the generation of ATP but does not depend on the protonmotive force. Unfortunately, studies with everted membrane vesicles failed to reveal the expected sodium pump; instead, the vesicles contained a conventional secondary Na+/H+ antiporter. We report here that everted membrane vesicles prepared in the presence of protease inhibitors retain an ATP-driven sodium transport system. The evidence includes the findings that (i) accumulation of 22Na+ by these vesicles is resistant to reagents that dissipate the protonmotive force but requires ATP and (ii) the vesicles contain a sodium-stimulated ATPase that is distinct from F1F0 ATPase, and whose presence is correlated with sodium transport activity. Sodium movements appear to be electroneutral and are accompanied by movement of H+ in the opposite direction. When membranes are incubated in the absence of protease inhibitors, a secondary Na+/H+ antiport activity emerges, possibly by degradation of the sodium pump. We suggest that S. faecalis expels Na+ by means of an ATP-driven primary transport system that mediates exchange of Na+ for H+. The Na+/H+ antiporter seen in earlier membrane preparation is an artefact of proteolytic degradation. PMID- 6283546 TI - Viral src gene products are related to the catalytic chain of mammalian cAMP dependent protein kinase. AB - The transforming protein sequences translated from the Rous avian and Moloney murine sarcoma virus src genes are shown to be related to the catalytic chain of bovine cAMP-dependent protein kinase (ATP:protein phosphotransferase, EC 2.7.1.37). The avian transforming protein, also a protein kinase, shows greatest homology with the bovine protein kinase in the carboxyl-terminal half, where the protein kinase activity is localized. Moreover, lysine occurs in the inferred transforming protein sequences at the position homologous with the proposed ATP binding lysine of the bovine protein kinase. This relationship is consistent with the hypothesis that the src genes originated in the host genomes, in which they are members of a superfamily of distantly related protein kinases that are normal constituents of mammalian cells. In the host, these sequences are much more highly conserved than in the viruses. PMID- 6283547 TI - Protease-catalyzed peptide bond formation: application to synthesis of the COOH terminal octapeptide of cholecystokinin. AB - This study of protease-catalyzed peptide synthesis reports the preparation of the COOH-terminal octapeptide amide of cholecystokinin. The octapeptide was assembled by chemical condensation of two tetrapeptide segments that had been synthesized through the concerted catalytic reactions of several proteases of different specificities. The resulting octapeptide derivative was subjected to catalytic transfer hydrogenation, followed by sulfation of its tyrosine residue and removal of the N alpha-protecting group. The homogeneous target peptide was obtained after purification via partition chromatography, gel filtration, and ion-exchange chromatography. The synthetic octapeptide stimulated amylase release from pancreatic acinar cells. PMID- 6283548 TI - Phosphorylation of the regulatory subunit of yeast cAMP-dependent protein kinase. AB - In vitro phosphorylation of the regulatory subunit of yeast cAMP-dependent protein kinase was studied. The cAMP-binding regulatory subunit (R subunit) can be multiply phosphorylated. Three distinct phosphorylation sites were inferred from the different ATP concentrations required for phosphorylation and from the presence of two discrete mobility shifts in NaDodSO4/polyacrylamide gel electrophoresis of the R subunit on phosphorylation. Limited tryptic digestion of the phosphorylated R subunit showed that a Mr 37,000 cAMP-binding peptide contained one of the phosphorylation sites and that a separate Mr 12,000 peptide contained another phosphorylation site. The yeast R subunit is therefore similar to the type II R subunit of mammalian origin, although it has a larger Mr (64,000 vs. 58,000) and is multiply phosphorylated. In vivo, both phosphorylated and unphosphorylated forms of the R subunit were found in cells grown in lactate or to stationary phase in 1.5% glucose, while cells grown in 5% glucose contained the unphosphorylated form. PMID- 6283549 TI - Simple model for the chemical potential change of a transported ion in active transport. AB - The mechanism for active transport of ions across a membrane probably involves two distinct conformational states of the transport protein, in which the binding sites for the transported ion face opposite sides of the membrane. It is likely that the binding affinity for the ion changes in synchrony with the change in site orientation, such that the affinity is high on the uptake side of the membrane and low on the discharge side. A structural model is proposed for the transmembrane portion of such a protein, based on the known multihelical structure of bacteriorhodopsin. This structure is well adapted to a cyclical alternation between two conformations that differ simultaneously in orientation and binding affinity. No unfolding of the helices or other significant alterations in secondary structure is required. The model is explicitly intended as a hypothetical representation of the E1 and E2 states of ATP-driven Na+,K+ and Ca2+ pumps. PMID- 6283550 TI - Novel free radicals in synthetic and natural pheomelanins: distinction between dopa melanins and cysteinyldopa melanins by ESR spectroscopy. AB - Synthetic pheomelanins from enzymic oxidation of the 3,4-dihydroxyphenylalanine (dopa) derivative 5-S-cysteinyldopa have been examined by ESR spectroscopy. These alkalisoluble polymers contain a novel kind of free radical that is spectroscopically distinct from that found in eumelanins. Delocalization of the unpaired electron onto a nitrogen atom and the ability of the radical to chelate complexing metal ions strongly suggest an o-semiquinonimine structure. The synthetic pheomelanin was compared with natural red pigments extracted from human red hair and from red chicken feathers. Spectroscopically, the chicken feather pheomelanin is almost identical to synthetic cysteinyldopa pheomelanin. In contrast, the pigment from red hair has a major spectral component very similar to that found in dopa melanin, with a smaller component corresponding to that found in cysteinyldopa melanin. PMID- 6283552 TI - Human gamma-trace, a basic microprotein: amino acid sequence and presence in the adenohypophysis. AB - The amino acid sequence of human gamma-trace, a basic microprotein without known function, was determined by automated Edman degradations of the carboxymethylated polypeptide chain and of fragments obtained by cyanogen bromide treatment and tryptic digestion after blocking of lysine residues. The single polypeptide chain contained 120 residues, and the calculated Mr was 13,260. A proline residue at position 3 was partly hydroxylated. The presence of gamma-trace in a significant proportion of the cells in the anterior lobe of simian and human pituitary glands was demonstrated by immunohistochemical procedures with a rabbit antiserum against human gamma-trace. The tissue localization and amino acid sequence of gamma-trace indicated that this protein is connected with the peptidergic gastroenteropancreatic neuroendocrine system. PMID- 6283551 TI - Staphylococcal plasmids that replicate and express erythromycin resistance in both Streptococcus pneumoniae and Escherichia coli. AB - Plasmid pSA5700 from Staphylococcus aureus coding for erythromycin (EmR) and chloramphenicol (CmR) resistance was transformed into Streptococcus pneumoniae. High-copy-number and EmR constitutive mutants of this plasmid were isolated. Transformation frequencies in S. pneumoniae as high as 70% were obtained with a constitutive plasmid as donor DNA, into a recipient cell containing a resident, inducible, high-copy-number plasmid. With the aid of these high frequencies, the site of constitutive mutations could be mapped via a simple marker rescue technique that uses purified restriction endonuclease-generated fragments. One of the EmR constitutive mutants, pFB9, a plasmid originating from a Gram-positive host, was shown to replicate and express EmR and CmR in a Gram-negative organism, Escherichia coli. Four derivatives of pFB9 containing large (0.6-0.9 megadalton) insertion sequences that arose spontaneously in E. coli demonstrated unusual transforming activity, as well as enhanced EmR, in E. coli. The inserted elements mapped to the region in front of the EmR gene. Three of these inserted elements had the size and restriction patterns of insertion sequence IS1, IS2, and IS5. Plasmid pFB9 and derivatives are useful for isolation of new insertion sequences and for comparison of gene expression and illegitimate recombination between Gram positive and Gram-negative species. PMID- 6283553 TI - Induction of sarcomas in athymic mice. AB - During the course of serial passage of 50 human xenografts in the athymic mouse over a period of 5 years we have observed two cases of induction of sarcomas in the murine stromal tissue associated with the human xenografts. Both times the growth of the murine sarcomas overtook that of the human xenograft. This change was monitored by analysis of the lactate dehydrogenase isozyme profile and histology of each passage of the human xenografts in the athymic mice. The two murine sarcomas were subsequently established in tissue culture. The sarcoma cell lines were found to be malignant by morphological and growth characteristics and were tumorigenic. They contained large amounts of murine leukemia virus when assayed for reverse transcriptase activity by infection of mouse SC-1 cells and BALB/c and NIH Swiss fibroblasts with filtered supernates, and some type C virus particles were observed by electron microscopy in tumor tissues. However, we were unable to demonstrate the presence of murine sarcoma virus by in vitro transformation of fibroblasts or sarcoma formation in vivo will cell free filtrates. Preliminary biochemical data indicate that the sarcomas are extremely high in plasma membrane ATP phosphohydrolase. PMID- 6283554 TI - Cyclic GMP mimics the muscarinic response in Xenopus oocytes: identity of ionic mechanisms. AB - Acetylcholine (AcCho) elicits four distinct membrane responses in Xenopus oocytes; the responses can be studied by using the voltage clamp technique. The fastest of the responses, a transient inward current (D1 response), is muscarinic, being evoked by oxotremorine and blocked by atropine but not by curare or hexamethonium. The action of AcCho is cooperative, three transmitter receptor complexes being required to cause a membrane conductance change, and the dose-response curve in most cases can be fitted by an equation assuming the existence of two binding sites with an affinity ratio of about 11. Guanosine 3',5'-cyclic monophosphate and the 8-bromo and dibutyryl derivatives cause a response similar to D1 in both its time course and the underlying ionic mechanism. The nucleotide-generated response has a smaller amplitude than the AcCho-generated D1. PMID- 6283555 TI - Correlation of regional brain metabolism with receptor localization during ketamine anesthesia: combined autoradiographic 2-[3H]deoxy-D-glucose receptor binding technique. AB - LKB film autoradiography of 2-]3H]deoxy-D-glucose uptake shows that ketamine, administered in anesthetic doses, alters the pattern of metabolic activity in rat hippocampus. The labeled metabolic marker can be washed out of the slide-mounted tissue sections by preincubation to permit in vitro autoradiography of drug and neurotransmitter receptors in the same animal. In this way, opiate and phencyclidine receptor distributions may be correlated with patterns of glucose utilization in adjacent sections. If the observed relative enhancement of 2-deoxy D-glucose uptake in the stratum moleculare of hippocampus reflects elevated metabolism in nerve terminals there, then the binding of ketamine to phencyclidine receptors on neurons in distant afferent sites, such as entorhinal cortex, may initiate the physiologic and metabolic effects. PMID- 6283556 TI - Age-dependent rotavirus-enterocyte interactions. PMID- 6283557 TI - Solid-phase radioimmunoassay of tonin in extracts of submandibular glands of rats treated chronically with isoproterenol. PMID- 6283559 TI - Angiotensin converting enzyme in cultured fibroblasts in Gaucher and Niemann-Pick diseases. PMID- 6283558 TI - Phosphorylation of membrane proteins by protein kinase in a smooth muscle plasma membrane fraction. PMID- 6283561 TI - Differential susceptibility of human cells to transformation by murine and avian sarcoma viruses. PMID- 6283560 TI - Effects of L-dopa and L-tyrosine on adrenergic transmission in the canine saphenous vein. PMID- 6283562 TI - A pharmacological approach to the structure of the Na channel in squid axon. PMID- 6283563 TI - The interaction of nerve growth factor with peripheral neurons: properties of the plasma membrane receptor. PMID- 6283564 TI - Gating charge movements and kinetics of excitable membrane proteins. PMID- 6283565 TI - Chemical modification of excitable membranes. AB - We have presented the effects of several chemical modification procedures on the properties of excitable membranes. The approach represents a middle ground between the biochemical approach of total separation of the proteins responsible for excitability and the biophysical approach in which only the physical parameters associated with the proteins are described. The results from investigations using group specific probes have begun to provide some picture, albeit a sketchy one, of the functional architecture of the proteins associated with excitability. As such, the investigations provide important information about necessary residues and their conformational arrangement in the intact protein. This knowledge will be especially useful in understanding excitability once the primary amino acid structure of the excitability proteins is determined. PMID- 6283567 TI - Rotation induced by intranigral phenobarbital: evidence of barbiturate GABAergic activity. PMID- 6283566 TI - Dynorphin-(1-13), dopamine and feeding in rats. AB - Intraventricular administration of the dopamine agonist, bromergocryptine, reliably induces feeding over a narrow dose range with a bell-shaped curve. Bromergocryptine (80 micrograms) induced feeding is inhibited by the dopamine antagonist, haloperidol (0.5 mg/kg) and the opiate antagonist, naloxone (10 and 1 mg/kg). The leucine-enkephalin containing opioid peptide, dynorphin-(1-13) induces feeding which is inhibited by haloperidol (0.5 and 0.1 mg/kg) and by naloxone (1 mg/kg). Of the common satiety factors tested only bombesin (10 micrograms/kg subcutaneously) inhibited both dynorphin-(1-13) and bromergocryptine induced feeding. Cholecystokinin-octapeptide (10 and 20 micrograms/kg, subcutaneously), thyrotropin-releasing hormone (10 and 20 micrograms), ICV) and calcitonin (1 unit, ICV) all failed to inhibit dynorphin-(1 13)-induced feeding. Calcitonin and CCK-8 but not TRH inhibited bromergocryptine induced feeding. These studies have demonstrated the close interaction between dopaminergic an dopiate systems in the regulation of food intake. The concept of dopamine being primarily responsible for the initiation of chewing behavior and the opiates regulating food ingestion is compatible with the observations reported here. PMID- 6283568 TI - Naloxone alters alcohol drinking induced in the rat by tetrahydropapaveroline (THP) infused ICV. AB - In male rats of the Sprague-Dawley or Long-Evans strain, intracerebroventricular (ICV) cannulae were implanted permanently using stereotaxic techniques. Tetrahydropapaveroline (THP) was infused ICV for up to 14 days either chronically around the clock or acutely once per day in doses previously found to induce an abnormally high intake of alcohol. During these periods, alcohol preference for individual rats was determined by a self-selection procedure in which the concentration of alcohol was increased from 3 to 30% on each day of a 12-day interval. Those rats which displayed a substantial increase in their intake of alcohol were selected for naloxone treatment and subsequently assigned a fixed concentration of alcohol at which maximum consumption occurred. Naloxone was administered subcutaneously two to six times per day for three consecutive days in total daily doses ranging from 1.5 to 3.0 mg/kg. Each rat served as its own control and was given 0.9% saline isovolumetrically according to the same temporal schedule. Naloxone generally suppressed alcohol intake in animals by 20% to 45%, but the reduction in drinking depended upon the injection regimen as well as the prior level of alcohol consumption. In "high drinking" rats, the mean alcohol intake of 6.2 g/kg/day was reduced to 3.7 g/kg/day by naloxone whereas in "low drinkers" the mean intake of 3.5 g/kg/day was suppressed to 2.8 g/kg/day by the opiate antagonist. These results further support the suggestion of a possible opiate receptor link in the pathogenesis and maintenance of aberrant drinking of alcohol, the mechanism of which may involve the endogenous action of an amine aldehyde condensation product in the brain. PMID- 6283569 TI - Isolation rearing decreases opiate receptor binding in rat brain. PMID- 6283570 TI - Presence of adrenocorticotropin-potentiating activity in a calcitonin preparation derived from porcine thyroid glands. AB - A calcitonin preparation derived from porcine thyroid glands exhibited potentiating activity on the steroidogenic action not only on ACTH-(1-24) but also of ACTH-(1-18) in isolated rat adrenal cells in vitro. This activity was demonstrated in various molecular weight fractions resulting from gel filtration of the preparation. This preparation also contained minute amount of immunoreactive and biologically active ACTH. PMID- 6283571 TI - Physical mapping of homologous segments of mitochondrial episomes from S male sterile maize. PMID- 6283572 TI - A physical map of plasmid pDU1 from the cyanobacterium Nostoc PCC 7524. PMID- 6283573 TI - Octopine Ti-plasmid deletion mutants of agrobacterium tumefaciens with emphasis on the right side of the T-region. PMID- 6283574 TI - Analysis of linear plasmids isolated from Streptomyces: association of protein with the ends of the plasmid DNA. PMID- 6283575 TI - Characterization of plasmids from plant pathogenic pseudomonads. PMID- 6283576 TI - Growth hormone response to clonidine as a probe of noradrenergic receptor responsiveness in affective disorder patients and controls. AB - The growth hormone (GH) response to the alpha-adrenergic agonist clinidine was blunted in 19 depressed patients compared to 20 controls. The difference remained significant when age- and sex-matches pairs of patients and controls were compared from this sample, either including or excluding subjects with elevated GH baseline levels. Plasma levels of free 3-methoxy-4-hydroxyphenyl-glycol (MHPG) were assayed in blood samples drawn just before the clonidine infusion. A modest negative correlation was found between the plasma MHPG values and the magnitude of the GH responses to clonidine, although baseline plasma MHPG levels were not significantly different between patients and controls. The diminished GH response to clonidine observed suggests that many depressed patients may have decreased alpha-adrenoreceptor responsiveness. Decreased responsiveness may in some cases be associated with relatively increased indices of presynaptic noradrenergic availability. Such a model might have implications for understanding the functional status of the noradrenergic neurotransmitter system in depressed patients and the possible subtyping of affective disorder patients. PMID- 6283577 TI - Pre-exposure to flavor and conditioned taste aversion: amphetamine and lithium reinforcers. PMID- 6283578 TI - Biological rhythms in development. PMID- 6283579 TI - Absence of naloxone sensitivity in obese humans. AB - Studies in rodents suggest the possibility of an association between elevated endogenous opiate activity and overeating and obesity. One measure of elevated opiate activity is sensitivity to the opiate antagonist naloxone. Seven massively obese human subjects showed no subjective or physiological sensitivity to large intravenous doses of naloxone. This finding fails to support a relationship between elevated endorphin activity and human obesity. PMID- 6283580 TI - [Rare case of particularly extensive odontodysplasia (II)]. PMID- 6283581 TI - [Progress in the field of synthetic fillings. Filling materials with powdered silicon dioxide as an inorganic filler]. PMID- 6283582 TI - [Comparative study of the sensitivity to sodium cefotaxime of 100 bacterial strains isolated from healthy volunteers and 100 strains isolated from "high risk" patients (author's transl)]. PMID- 6283583 TI - Alteration in thermal sensitivity of chinese hamster cells by D2O treatment. PMID- 6283585 TI - [Cyclic nucleotides and radioresistance. 6. Modification of the effect of cyclic nucleotides on the radioresistance of E. coli with changes in the composition of the growth medium]. PMID- 6283586 TI - [Thyroid neoplasms (diagnosis, therapy, after care). Abstracts]. PMID- 6283584 TI - [Mechanism of chromatin degradation in the thymocytes of irradiated rats. 3. Postradiation activation of autolytic disintegration of DNA in isolated thymocyte nuclei]. PMID- 6283587 TI - [Experimental studies with new embolization material (cyanoacrylate-contrast medium mixture). Part 1 (author's transl)]. PMID- 6283588 TI - [Computed tomography of ovarian tumours-progress in diagnosis (author's transl)]. PMID- 6283589 TI - [Computed Axial tomography in patients with abdomino-perineal excision. A radiological and clinical control (author's transl)]. AB - From 1978 to 1980 a total of 119 patients with carcinoma of the rectum were treated by abdominoperineal excision at Erlangen University Surgical Clinic. 66 of these patients were examined by CAT scan in a follow-up study. Local recurrence was found in nine patients. A radical second operation was useful in one case only. Our findings show, that only postoperative check-ups by CAT scan can detect new tumor growth at an early state. The prognostic groups used in Erlangen are helpful in defining the intervals for postoperative examinations by CAT scan. PMID- 6283590 TI - [Radiology of the infantile hand as a mirror of clinical syndromes (author's transl)]. AB - Significant radiographic findings of the pediatric hand are described as far as they are useful for the diagnosis of a suspected clinical syndrome. Normal variants that can simulate pathological findings are demonstrated. Deviations of skeletal age from chronological age and their clinical significance are discussed. Diagnostically useful malformations (hexadactyly, syndactyly, brachyphalangy, cone shaped epiphyses and anomalies of the thumb) are presented in the context of typical cases of selected syndromes. The importance of establishing a correct diagnosis in these cases is stressed with regard to individual prognosis of the patient and genetic counselling of the family. PMID- 6283591 TI - [Radiologic-pathologic correlations in tumors and tumor-like lesions of the hand (author's transl)]. AB - Since 1972, 267 tumors or tumor-like lesions of the hand in 257 patients were examined at the bone tumor registry by the Swiss Society of Pathology in Basel, Switzerland. This is 9% of a total of 2,800 bone tumors submitted. An analysis revealed 254 benign tumors or tumor-like lesions and 13 malignant tumors, among them 2 metastases of tumors of unknown origin. As expected, enchondromas (n=81) and pigmented villo-nodular synovitis of the tendon sheath (n=104) were the most frequent tumors. All other tumors or tumor-like lesions appeared only as solitary cases or in small numbers. The radiologic characteristics of some of the lesions are reviewed and their histology is presented. Furthermore, some important points of differential diagnosis are discussed. PMID- 6283592 TI - Pathways of tumor spread through the lung: radiologic correlations with anatomy and pathology. AB - The pathways of tumor spread through the lung are described and their significance for radiographic interpretation is illustrated. A key to understanding the spread of bronchogenic carcinoma is the realization that although the normal flow of lymph in the pulmonary lymphatics is centripetal, lymphatic obstruction can cause reversal of flow. As a result, tumor cells are commonly carried centrifugally to the periphery in lymphatics or the connective tissue around them, and remote pleural involvement, secondary parenchymal masses, or satellite nodules may develop. Failure to appreciate peripheral spread of tumor has negative consequences for tumor staging, surgery, and radiotherapy. In the absence of hilar node involvement causing obstruction, long line shadows more than 0.5 inch (1.25 cm) in length proximal to a peripheral mass very infrequently represent tumor. PMID- 6283593 TI - The angiomatous polyp and the angiofibroma: two different lesions. AB - Five cases of angiomatous polyps that were initially interpreted as angiofibromas are presented. Although they are similar to angiofibromas, angiomatous polyps have a distinctly different pattern of growth on computed tomography and a different angiographic appearance. In addition, they are more easily extirpated at surgery than are angiofibromas. PMID- 6283594 TI - Proton nuclear magnetic resonance imaging of regionally ischemic canine hearts: effect of paramagnetic proton signal enhancement. AB - In a study to evaluate the potential of proton nuclear magnetic resonance (NMR) imaging with and without manganese contrast with and without manganese contrast enhancement for detecting acute myocardial infarction, 12 dogs underwent 90 minute occlusion of the left circumflex coronary artery. Transverse-section NMR images of the excised, nonbeating heart were obtained at 1-cm intervals using the steady-state-free-precession (SSFP) technique. All NMR images revealed detailed structure of the heart. The three hearts without manganese showed no difference in intensity between the normal and the ischemic posterior regions, whereas those with manganese demonstrated a clearly demarcated zone of reduced signal intensity consistent with the ischemic zone. It is concluded that high-resolution tomograms of the excised canine myocardium can be obtained using proton NMR imaging. With the SSFP imaging technique, proton signal enhancement with manganese infusion is necessary to differentiate between ischemic and nonischemic myocardium after 90 minutes of coronary occlusion. PMID- 6283595 TI - [Histochemical studies of carbohydrate and lipid metabolism in human liver biopsies]. PMID- 6283596 TI - Lysozyme (muramidase) and alpha 1-anti-chymotrypsin as immunohistochemical tumour markers. AB - Since lysozyme and alpha 1-anti-chymotrypsin are constituents of normal histiocytes, their value as tumor cell markers in histiocytes neoplasias has been investigated using the indirect immunoperoxidase method and commercially available specific antisera on formaldehyde-fixed, paraffin-embedded 5 micrometers sections after pretreatment with pronase. The distribution of both markers was determined in 35 cases of malignant fibrous histiocytoma (MFH) and in 13 cases of malignant histiocytosis (MH). In 12 cases of MH both markers were found whereas in MFH alpha 1-antichymotrypsin was demonstrated in 26 and lysozyme in 16 cases only. In general, the staining for alpha 1-anti-chymotrypsin was more intense than the staining for lysozyme. A negative reaction does not exclude the possibility of MH or MFH. The presence of both constituents in tumours, however, can be considered as indicative of histiocytogenic origin and both can be useful markers for distinguishing histiocytic neoplasias from other tumours. PMID- 6283597 TI - Chemomorphology of liver parenchyma. Qualitative histochemical distribution patterns and quantitative sinusoidal profiles of G6Pase, G6PDH and malic enzyme activity and of glycogen content. PMID- 6283598 TI - Persistent, slow and latent viral infections. PMID- 6283600 TI - Epstein-barr-virus nuclear antigen. PMID- 6283599 TI - Cell growth transformation by herpes simplex virus. PMID- 6283601 TI - Taxonomy and nomenclature of viruses, 1982. PMID- 6283602 TI - Cytomegalovirus infection in transplant patients. PMID- 6283603 TI - [Gastrointestinal transit in the chicken using 198Au-colloid as a marker (author's transl)]. AB - Gastrointestinal transit in the chicken was investigated by using 198Au-colloid as a marker. Gastrointestinal transit was determined following administration of a 198Au-colloid solution (370 kBq (10 microCi), 0.5 ml) into the proventriculus by measuring the distribution of radioactivity in the gastrointestine. Most of 198Au-colloid administered into the proventriculus was transferred instantly to the gizzard and subsequently, some part of it quickly to the duodenum and the upper segment of the jejunoileum. A considerable quantity of 198Au-colloid in the duodenum, the upper and middle segment of the jejunoileum regurgitated to the gizzard. 198Au-colloid transferred to the caecum was retained for a long time (more than 48 hours) in these segments. A part of 198Au-colloid was found in the feces 90 to 120 minutes after administration into the proventriculus and the greater part of it was excreted into the feces 24 to 48 hours. Subcutaneous administration of acetylcholine (2 mg/kg) increased the gastrointestinal transit but atropine (2 mg/kg) decreased. PMID- 6283604 TI - [Mesenteric glomic tumor]. PMID- 6283605 TI - [Differential diagnosis of nasopharyngeal cancer]. PMID- 6283606 TI - [Comparative study of the hypouricemic effects of calcitonin and sulfinpyrazone]. PMID- 6283607 TI - [Eaton-Lambert syndrome. Apropos of 4 cases]. PMID- 6283608 TI - [Emphysematous cholecystitis. Report of 2 cases]. PMID- 6283609 TI - [Ortho- and paraendocrine pancreatic apudomas (report of 4 personal cases)]. PMID- 6283610 TI - Oronasal challenge of fattening pigs after vaccination with an inactivated Aujeszky's disease vaccine. AB - Groups of pigs from vaccinated and unvaccinated sows were vaccinated once or twice between the ages of eight and 20 weeks with a commercial inactivated, oil adjuvanted Aujeszky's disease virus vaccine. Pigs were challenged by the oronasal route when 22 to 27 weeks old. Pigs from unvaccinated sows developed neutralising antibodies after vaccination but no seroconversion was detected in eight-week-old pigs or in 80 per cent of 15-week-old pigs from vaccinated sows. Challenge resulted in severe disease and weight loss in control pigs. In vaccinated animals the duration and severity of clinical signs and the amount of weight lost decreased with increasing serum neutralisation titres. The results indicate that parenteral vaccination at weaning with the vaccine described will not protect pigs at slaughter age against infection and disease, particularly if they were born from seropositive mothers. PMID- 6283612 TI - Incidence of rotavirus in artificially reared pigs and some effects of diarrhoea on the physiology and histology of the gastrointestinal tract. AB - Pigs reared on a milk substitute from two days old often developed diarrhoea, but suckled littermates remained healthy. Only in a few pigs was diarrhoea associated with the presence of rotavirus. Rotavirus was also present in some healthy pigs, and was associated with a reduction in villus length. Pigs with diarrhoea usually had an increased amount of digesta in the stomach and a reduction in lactase activity in the small intestine but villus length was unchanged. There was no evidence of lactose malabsorption. PMID- 6283613 TI - Diarrhoea in dairy calves reduced by feeding colostrum from cows vaccinated with rotavirus. AB - Forty-two dairy calves remained with their dams for two days after birth, and then were removed to a calf rearing shed. Calves were allocated to three groups for the next 14 days, and received twice daily either whole milk, whole milk with a 10 per cent supplement of pooled normal bovine colostrum or whole milk with 10 per cent supplement of colostrum from cows vaccinated with rotavirus. A natural outbreak of diarrhoea occurred, affecting 28 of the 42 calves. Feeding immune colostrum delayed the onset of diarrhoea, and reduced its incidence, duration and severity. Live weight gains were consequently improved. The group fed normal colostrum had diarrhoea intermediate in severity between that of control calves and those fed immune colostrum. The aetiology of the diarrhoea was complex, with calves excreting rotavirus, enteropathogenic Escherichia coli and cryptosporidia. PMID- 6283614 TI - Duration of immunosuppression caused by a vaccine strain of infectious bursal disease virus. AB - The depression of response of Brucella abortus strain 19 caused by an infectious bursal disease vaccine virus given to chicks at one day old was shown to persist for four weeks. Histological examination of the bursa of Fabricius showed a gradual repopulation by bursal lymphocytes, after initial damage, concomitant with the development of a humoral response. PMID- 6283611 TI - Efficacy of an inactivated feline calicivirus (FCV) vaccine against challenge with United Kingdom field strains and its interaction with the FCV carrier state. AB - The efficacy of an inactivated vaccine derived from feline calicivirus (FCV) strain FS2 was assessed against challenge with three UK field strains of FCV. The mean clinical score, calculated on the number of signs recorded per day over 21 days after challenge, was lower for vaccinated cats when compared to unvaccinated animals though the difference was not statistically significant. All cats excreted FCV throughout the three weeks following challenge and there was no difference in the number of days of virus shedding during this period between vaccinated and unvaccinated animals. The development of FCV serum neutralising antibody titres following vaccination and challenge was recorded. In the second part of the study the ability of vaccinated and challenged cats to become FCV carriers and then infect susceptible in-contact animals was demonstrated. PMID- 6283615 TI - [Problems of vaccination against herpes viruses (author's transl)]. PMID- 6283616 TI - Apolipoprotein E (role in lipoprotein metabolism and pathophysiology of hyperlipoproteinemia type III). PMID- 6283617 TI - Pregnancy-specific beta 1-glycoprotein in complicated early pregnancy and trophoblastic disease. PMID- 6283619 TI - [So-called sporadic ulcero-mutilating acropathy: clinical study of 10 cases. P I]. PMID- 6283618 TI - Solid-phase radioimmunoassays for Epstein-Barr virus associated antigens. PMID- 6283620 TI - [Nitrate and nitrite content in selected vegetables]. PMID- 6283622 TI - [Serological studies in research on arbovirus antibodies in the human population of the Ribeira Valley region. II - Survey of patients of Pariquera-Acu Regional Hospital, 1980]. PMID- 6283621 TI - [Hypophosphoremic osteomalacia of connective tissue tumors]. AB - The authors undertake a general review of the association between hypophosphoraemia and connective tissue tumour, based upon three personal cases and 27 cases of benign connective tissue tumours, as well as cases of hypophosphoraemia related to malignant tumours or to diffuse dysplasia of connective tissue origin, collected from the literature. This syndrome is distinguished from hypophosphoraemia induced by other tumours (myeloma, carcinoma of the prostate) which are based upon different mechanisms. Hypophosphoraemia, associated with a fall in plasma levels of 1-25 (OH)2 D3 by inhibition of renal 1 alpha hydroxylase, suggests the existence of a complex tubular deficit. Removal of the tumour, most often vascular and intra- or para-osseous, results in rapid normalisation of laboratory then radiological and clinical abnormalities. The physiopathology of the syndrome remains very mysterious. It may be likened to certain tubulotoxic syndromes due to cadmium and in particular to maleic acid. However no precise data yet exists regarding any possible abnormal tumour secretion. In practice, any case of hypophosphoraemic osteomalacia requires investigation to locate a possible tumour of connective tissue, and this all the more so when it is accompanied by very low plasma levels of 1-25(OH)2 D2. PMID- 6283623 TI - [Effects of acute hypoxia upon the concentration of cytochromes in the brain and the liver of differently aged rats (author's transl)]. PMID- 6283624 TI - Transferrin endocytosis in reticulocytes: an electron microscope study using colloidal gold. AB - The endocytosis of transferrin by rabbit reticulocytes was investigated by electron microscopy using transferrin labelled with colloidal gold. This complex was shown to bind and donate its iron to the cells in a manner comparable to native iron-transferrin. After incubation at 37 degrees C approximately 70% of the transferrin-gold particles were located within endocytotic vesicles. Treatment of the cells with pronase, EDTA, metabolic inhibitors and heating to 46 degrees C inhibited the endocytosis. The uptake of colloidal gold complexes of albumin and concanavalin A were compared with that of transferrin. Little endocytosis of these proteins was observed. It is concluded that colloidal gold is a suitable label for the investigation of transferrin endocytosis and that the endocytosis is specific for transferrin and sensitive to the action of reagents which damage membrane receptors, alter membrane function or impair cell metabolism. PMID- 6283625 TI - Single drug therapy in non-seminomatous germinal cell tumours of the testicle. AB - The outcome for 84 patients in different stages of non-seminomatous germinal cell tumours of the testicle and treated with Mithramycin as a single agent has been evaluated. The survival rate in patients with advanced disease was lower than that obtained by more aggressive chemotherapy and, therefore, in these stages Mithramycin therapy is not recommended. On the other hand, the survival rate in patients with low-stage disease was superior to that in other series of patients subjected to bilateral lymphadenectomy without subsequent adjuvant chemotherapy. In the present study a unilateral gland dissection with preservation of a normal sexual function was performed in the vast majority of patients. Single-drug chemotherapy should preferably be used only as adjuvant treatment in low-stage non-seminomatous germinal cell testicular tumours. PMID- 6283626 TI - [Angiotensin receptors and antagonists (author's transl)]. PMID- 6283627 TI - [Characteristics of the responses of cardiac alpha-adrenoceptor and its functional significance (author's transl)]. PMID- 6283628 TI - [Recent progress in enkephalin research (author's transl)]. PMID- 6283629 TI - Tampon associated toxic shock syndrome. PMID- 6283630 TI - Indoor airborne asbestos pollution: from the ceiling and the floor. AB - Electron microscopic measurements of the concentrations of airborne asbestos were carried out inside and outside an office building having ceilings sprayed with a crocidolite-containing material and floors covered with vinyl-chrysotile tiles. Under normal conditions in this building, constructed 10 years ago, the two asbestos-containing materials released fibers into the air. This is the first measurement of elevated (up to 170 nanograms per cubic meter) concentrations of indoor airborne asbestos associated with the weathering of asbestos floor tiles during their service life. Asbestos flooring is used in a large number of buildings and represents the third largest use of asbestos fibers in the United States and in Europe, ranking after roofing and asbestos-cement pipe. PMID- 6283631 TI - Nucleotide sequence of the transforming gene of avian myeloblastosis virus. AB - Avian myeloblastosis virus is defective in reproductive capacity, requiring a helper virus to provide the viral proteins essential for synthesis of new infectious virus. This virus arose by recombination of the nondefective helper virus and host cellular sequences present within the normal avian genome. These latter sequences are essential for leukemogenic activity. The complete nucleotide sequence of this region is reported. Within the acquired cellular sequences there is an open reading frame of 795 nucleotides starting with the initiation codon ATG (adenine, thymine, guanine) and terminating with the triplet TAG. This open reading frame could code for the putative transforming protein of 265 amino acids with a molecular weight of approximately 30,000. PMID- 6283632 TI - Corticotropin-releasing factor stimulates secretion of melanocyte-stimulating hormone from the rat pituitary. AB - Administration of synthetic ovine corticotropin-releasing factor led to rapid, parallel increases in adrenocorticotropin and alpha-melanocyte-stimulating hormone concentrations in rat plasma. Prior treatment with dexamethasone almost completely blocked the adrenocorticotropin response but not the increase in melanocyte-stimulating hormone. These data demonstrate that corticotropin releasing factor is a potent stimulator not only of adrenocorticotropin secretion from the corticotrophs of the anterior pituitary gland but also of peptide secretion from the intermediate lobe. Such data suggest that melanocyte stimulating hormone and beta-endorphin play a role in the physiological response to stress. PMID- 6283633 TI - Rat pro-opiomelanocortin contains sulfate. AB - Intermediate lobes isolated from rat pituitary glands incorporated [35S]sulfate into pro-opiomelanocortin and other adrenocorticotropic hormone-containing peptides. Incubation of intermediate lobes in medium containing the arginine analog canavanine inhibited the cleavage of pro-opiomelanocortin into smaller products. Pro-opiomelanocortin that accumulated in the presence of canavanine was also sulfated. PMID- 6283634 TI - Transcriptional control signals of a eukaryotic protein-coding gene. AB - Transcriptional control signals of a model eukaryotic protein-coding gene have been identified by a new procedure of in vitro mutagenesis. This method allows small clusters of nucleotide residues to be substituted in a site-directed manner without causing the addition or deletion of other sequences. Transcription assays of a systematic series of these clustered point mutants have led to the identification of three distinct control signals located within the 105 nucleotide residues immediately upstream from the point where transcription begins. PMID- 6283635 TI - Immunotherapy of metastases enhances subsequent chemotherapy. AB - In many multimodal therapies of cancer, postsurgical chemotherapy is administered before immunotherapy for treatment of micrometastatic disease. This sequence may not be the most efficacious. Experiments in which strain 2 guinea pigs bearing syngeneic L10 hepatocarcinomas were given immunotherapy showed that infiltrating immune effector cells not only were tumoricidal but disrupted the characteristically compact structure of metastatic foci. When cytotoxic drugs were administered at the peak of this inflammatory response, the survival rate of the guinea pigs increased significantly. We conclude that postsurgical immunotherapy can enhance the effect of cytotoxic drugs administered subsequently. PMID- 6283636 TI - Gene scanning with block mutations. PMID- 6283637 TI - Osmotic swelling of phospholipid vesicles causes them to fuse with a planar phospholipid bilayer membrane. AB - Fusion of phospholipid vesicles with planar bilayer membranes occurs if the vesicles that contact the planar membrane swell osmotically after the replacement in their medium of an impermeant solute by a permeant one. This finding directly demonstrates that osmotic swelling is a driving force for vesicle-planar membrane fusion. The method used to induce vesicle swelling and fusion may have relevance for biological systems. PMID- 6283638 TI - Eumelanins and pheomelanins: characterization by electron spin resonance spectroscopy. AB - Synthetic dopa melanin and cysteinyldopa melanin have different electron spin resonance spectra. Data are reported for mixtures of these melanins and for dopa cysteinyldopa copolymers, which are spectroscopically similar. A simple parameterization of the spectra allows estimation of the relative amounts of (i) dopa melanin and cysteinyldopa melanin in mixtures and of (ii) dopa and cysteinyldopa incorporated into copolymers. Several natural eumelanins and pheomelanins have been characterized and shown to be copolymers. PMID- 6283639 TI - Repeated DNA still in search of a function. PMID- 6283640 TI - Active genes are sensitive to deoxyribonuclease I during metaphase. AB - The active exogenous murine leukemia virus sequences of mouse cells growing in culture are preferentially digested by deoxyribonuclease I in metaphase chromosomes. As determined by nuclear nick translation, all of the gene sequences of these cells active during interphase are in a deoxyribonuclease I-sensitive conformation during metaphase. This method of nick translation can therefore be used to label chromosomes in situ in order to visualize the active regions of the genome. PMID- 6283642 TI - [Carotid bruit due to increased blood flow without stenosis: report on two cases (author's transl)]. AB - Initial reaction to the discovery of a carotid bruit in an adult is that a local atheromatous lesion exists. This sign may arise, however, when more distal lesions are present, such as an angioma provoking an increased rate of blood flow. All patients presenting with neurological signs of a lesion in one hemisphere should, therefore, in the absence of contra-indications, be investigated by arteriography. PMID- 6283641 TI - [Lithium and diabetes insipidus. Theoretical and practical aspects. Therapeutic value of thiazide diuretics (author's transl)]. AB - Lithium is increasingly prescribed by psychiatrists for manic-depressive psychosis and other affective disorders. Therefore, psychiatrists should be familiar with lithium-induced nephrogenic diabetes insipidus. Management of this condition is discussed. Thiazide diuretics have a paradoxical antidiuretic effect in lithium-induced diabetes insipidus. For this reason, it is no longer warranted to contraindicate concomitant use of lithium and thiazide diuretics. PMID- 6283643 TI - [Computed tomography of the head in gunshot wounds. Thirty-three cases (author's transl)]. AB - The authors report a series of 33 cases of craniocerebral lesions secondary to projectiles injuries studied by means of CT scan. Ct scans demonstrate the path of the missile, destructions of deep cerebral parenchyma, dissections of with matter (intra-cerebral bubble) and reactive oedema. Prognostic incidence of CT is discussed. CT scan helps to choose the best therapeutic approach as regards each particular case. PMID- 6283644 TI - [Meningeal carcinomatosis in adults (author's transl)]. AB - Twenty cases of carcinomatous meningitis in adults, treated in the Gustave Roussy Institute between 1970 and 1980, are reviewed. In fourteen cases meningitis was secondary to breast cancer. Initial inflammation was present in 90% of the mammary tumors. Clinical features as well as biochemical and cytological findings in cerebrospinal fluid are described. The various therapeutic regimens using radiotherapy and intrathecal chemotherapy are analyzed in terms of survival and compared with other published studies. The most significant point is the necessity for early diagnosis which improves survival. However, prognosis for this metastatic disease is always very poor. PMID- 6283646 TI - [What should be said to a depressed patient's family? (author's transl)]. PMID- 6283645 TI - [Clinical trial of tiapride in patients with dyskinesia (author's transl)]. AB - Twenty-five patients with various forms of dyskinesia were given tiapride for three months. Maximal dosage was 900 mg per day. A double-blind trial of tiapride versus placebo showed significantly better results in the group given tiapride. The forms of dyskinesia which responded best to tiapride were the following: iatrogenic dyskinesia, tics (Gilles de la Tourette syndrome), and chronic chorea (Huntington disease). Patients with complex dyskinesia resulting from neonatal encephalopathy or vascular disease were not improved. The protocol used in l-dopa induced dyskinesia is described. Changes in dyskinesia and "on-off" effect following variations in tiapride and l-dopa dosage are detailed. An unequivocal, although minor, tiapride-induced parkinson syndrome was recorded in a few patients. No instances of tiapride-induced dyskinesia or akathisia were seen. The other side-effects were either psychic (depression, drowsiness, agitation) or endocrinologic (menstrual disorders, overeating, galactorrhea). PMID- 6283647 TI - [Viral etiology of aseptic lymphocytic meningitis. Twenty-four cases seen over a nine-year period (author's transl)]. AB - Among 208 cases of aseptic lymphocytic meningitis seen over a nine-year period, 24 were diagnosed as viral infections. Diagnosis was established by both isolation of the virus from one or several specimens (feces, pharyngeal mucus, CSF) and significant rise in specific antibody titres. The usual clinical and epidemiological characteristics of these benign infections were demonstrated in our series: prevalence in males; recrudescence in summer; infectious syndrome associated with an unequivocal, though moderately severe, meningeal syndrome, and occasionally with various visceral manifestations (exanthema, pharyngitis, acute respiratory disease, myalgia); clear CSF usually with pleiocytosis (relative but not absolute predominance of lymphocytes) and normal chemical findings. All the cases in our series were due to enteroviruses, Echo or coxsackie. Isolation of a poliovirus (type III) in one case deserves mention. Cases where clinical examination led to unequivocal diagnosis of mumps or herpes zoster were not included in our study. PMID- 6283649 TI - [Emergency diagnosis of intracranial hypertension with infectious signs (author's transl)]. PMID- 6283648 TI - [Herpes simplex encephalitis in neuro-surgery. Electro-encephalographic contribution to early diagnosis (author's transl)]. AB - The electroencephalographic (EEG) signs of acute necrotizing encephalitis are always focalized initially. In over 80% of the cases, the EEG shows local periodical activity with a characteristically short interval between repetitive paroxysms. This was demonstrated by the study of 17 personal observations in which herpes simplex virus was unequivocally identified as the causal agent. This EEG pattern belongs to the general group of "periodical activities". It is an early but transient finding. When observed in an acute clinical picture with fever and encephalic symptoms, it is a major diagnostic feature and should lead to immediate active therapy (neuro-surgery or chemotherapy). PMID- 6283650 TI - [Hypogonadism with spermatogenesis (fertile eunuch syndrome) (author's transl)]. AB - Hypogonadism with spermatogenesis (fertile eunuch syndrome) occupies a special nosologic situation because of it's clinical and paraclinical features. These include: eunuchism of testosterone's target tissues, contrasting with clinically normal testes; normal embryonic male differentiation; low serum testosterone concentrations; moderately decreased or normal LH serum concentrations which are coincident with low testosterone levels therefore reflecting at least relative LH insufficiency; normal FSH levels; normal LHRH response; variable clomifen response; poor spermatogenesis improved by HCG and/or testosterone, and in some cases by HMG; atrophic leydig cells. Although in very rare instances it may include anosmia, the fertile eunuch syndrome should be clearly distinguished from complete FSH and LH deficiency with or without anosmia. It must also be distinguished from the very uncommon isolated FSH deficiencies. The fertile eunuch syndrome probably originates in insufficient hypothalamic production of LHRH. The mechanism by which spermatogenesis is at least partly spared remains unknown. PMID- 6283651 TI - [The immunology of Crohn Disease (author's transl)]. AB - Although the incidence of Crohn disease seems to be increasing, it's etiology remains unknown. The search for immunological mechanisms has been extensively carried out with conflicting results. At the present, most studies provide evidence suggesting a possible immunological dysfunction. Another interesting point is the possible role of an oxygen dependant cytotoxic activity of phagocytes in inflammatory processes, and particularly in Crohn disease. PMID- 6283652 TI - [Cell-mediated immunity studied by skin tests in patients receiving intensive care. Prognostic value of repeated tests. Study of some factors predisposing towards anergy (author's transl)]. AB - Cell-mediated immunity was assessed by three skin tests (using tuberculin, candidin, and Varidase) in one-hundred patients in a medical intensive care unit. Anergy on admission was most often found after major blood loss and massive transfusion as well as in patients over sixty. For the 49 patients who were anergic on admission the mortality rate was 32%, against 12% for the 51 reactive subjects (p less than 0.01). Repetition of skin tests considerably improved their prognostic value. No deaths occurred among the 21 patients who were reactive on admission and throughout the course of their disease, or among the 16 patients who, after being anergic on admission, became reactive subsequently. Conversely, the survival rate was only 40% for the 22 patients who remained or became anergic. Our results confirm the value of skin tests for assessing cell-mediated immunity in patients receiving intensive care. Repeated tests allow early detection of high risk patients in whom fatal outcome, whether due to infection or not, is more frequent. Two factors which predispose to anergy are underlined: advanced age and massive transfusion after major blood loss. PMID- 6283653 TI - [Acute peritonitis as a complication of Crohn's disease of the colon (author's transl)]. AB - Acute peritonitis is a rare complication of Crohn's disease of the colon. We present the case of a young woman with peritonitis complicating an acute exacerbation of Crohn's disease of the colon. Seventeen other cases from the literature show that perforation may be the first manifestation of the disease or may occur during an acute exacerbation of a chronic form. With the presently limited experience, the optimal surgical treatment has not been defined. PMID- 6283654 TI - [Importance of gastroscopy in chronic uremic patients (author's transl)]. AB - In chronic uremic patient, two kinds of digestive troubles can occur: gastric or duodenal lesions secondary to an abnormal gastro-intestinal hormone excretion, and drug-induced disorders. Most of these manifestations were missed by usual barium meal and the use of routine endoscopic check-up can be particularly useful. We performed 70 gastroscopies in 67 chronic hemodialyze patients. Examinations were practiced immediately before dialysis with only a light sedation. We observed 39 lesions: 3 drug-induced gastritis, 15 erosive or pseudopolypoid inflammations of the polyric antrum, 2 gastric ulcers, 5 duodenal ulcers (two of them were linear), 2 peptic oesophagitis and 34 p. cent of our patients had marked duodenitis, most of these digestive abnormalities could be missed or overdiagnosed by using routine barium meal. In presence of the number and the therapeutic importance of these disorders, we think that a gastroscopy should be prescribed in three indications: uremic patients before the beginning of an hemodialyze treatment, chronic hemodialyze patients with unexplained anemia, check-up before renal transplantation where ulcerations are a major contra-indication. In such situations, gastroscopy seems an important progress for the management of this particular group of patients. PMID- 6283655 TI - [Deferentography under local anaesthesia in male sterility : technique and results (author's transl)]. AB - Deferentography was employed during medical and surgical investigations of 44 cases of male sterility. This technique is easily performed in out patients and appears to be inocuous. Radiological signs were normal in 31 cases, and it was possible to evaluate anatomical characteristics and to obtain precise measurements of the vasa deferentia, the ampulla, the seminal vesicles, and the ejaculatory canals. Abnormal findings, present in 13 cases, corresponded to bilateral deferens obstruction of either congenital (agenesis), or acquired (stenosis due to infection or inflammation) origin. The major limitation of this investigational method is that epididymal obstruction cannot be demonstrated. PMID- 6283656 TI - [Cholesterolemia and pyrexy (author's transl)]. AB - During pyretic diseases from any origin, total plasma cholesterol level decreases and its values are frequently below 1 g per liter. This observation could be expected because it conforms with the following theory : 1) cholesterol synthesis in an organism is proportional to consumed energy, 2) plasma cholesterol level is the lower as cholesterol synthesis is high. PMID- 6283657 TI - [Anti-JKa specificity : a rare variety of auto-immune hemolytic anemia (report of a case associated with myxoedema (author's transl)]. AB - The authors report the case of a patient with autoimmune hemolytic anemia, due to anti-KJa antibody, and associated with primary hypothyroidism. They discuss the possible relationship between these two conditions and emphasize the particular hematological pattern related to myxoedema. PMID- 6283658 TI - [Bilateral pneumothorax revealing a perforated peptic ulcer (author's transl)]. PMID- 6283659 TI - [Anemia with hypochromia and decreased serum iron in malignant tumours of the small intestine. Two case-reports (author's transl)]. AB - The authors report two cases of malignant tumours of the small intestine (one adenocarcinoma and one lymphoma). In both patients, longstanding anemia with hypochromia and decreased serum iron remained unexplained for some time. Iron was prescribed without definite diagnosis. These two cases demonstrate the extreme latency of these tumours and the necessity for detailed intestinal investigations in patients presenting with isolated hypochromic hyposideremic anemia. Among these investigations, doubla contrast roentgenographic study of the small intestine appears to be an adequate diagnostic procedure for disclosing even very small lesions. PMID- 6283660 TI - [Adenocarcinoma of the kidney. A series of two hundred and thirty-seven cases (author's transl)]. PMID- 6283661 TI - [Hemodynamic effects of intravenous isosorbide dinitrate in chronic heart failure (author's transl)]. AB - During cardiac catheterization, isosorbite dinitrate (ISDN) was given intravenously to ten patients with confirmed left ventricular failure (10 NOMC, 1 ischemic cardiopathy, 1 valvular disease). When the left ventricular filling pressure was above 15 mmHg and the systemic systolic pressure exceeded 100 mmHg, a 500 gamma (first half of patients) or 800 gamma (second half of patients) bolus of ISDN was given intravenously, followed by an infusion of 2.5, 5 or 10 mg ISDN per hour. In all patients, the functional tolerance was excellent ; the heart rate remained unchanged and the systolic systemic pressure decreased only slightly (10 %). Left ventricular filling pressures returned to normal in four patients, decreased significantly in three, and remained unchanged in three. The cardiac output increased significantly in four patients. The poor initial hemodynamic conditions and the doses given, subsequently considered insufficient especially during the first part of the study, probably explain why ISDN was only partly effective. PMID- 6283662 TI - [Accidental poisoning with liquid or solid caustic soda for domestic use: circumstances and cost (author's transl)]. AB - 524 records of patients seen in French anti-poison centers in 1978-1979 for accidental poisoning with caustic soda were reviewed. The accidents were usually due to liquid agents. The digestive lesions were particularly serious in children who were affected in more than two-thirds of the cases. The accident usually occurred while the caustic agent was being used. PMID- 6283663 TI - [Human red cell cytoskeleton: structure, functions, abnormalities (author's transl)]. AB - Associated with the red cell membrane cytoplasmic surface, the cytoskeleton is composed of a protein complex. This structure is a major element of membrane stability, shape and flexibility. Among the constitutive proteins, spectrin, red cell actin and protein band 4.1 are quantitatively and structurally the most important. Some elements, relative to structure, stoichiometry and interactions between these proteins begin to be understood, specially owing to in vitro studies of reassembling cytoskeleton structures from purified constitutive elements. Besides, the nature of some interactions between the cytoskeleton and the red cell membrane has been specified. Nevertheless, it is not possible at the present time to establish an architectural picture of the cytoskeleton anatomy, and the factors actually regulating this structure physiology are not characterized. Understanding of the normal cytoskeleton will perhaps be helped by studies of abnormal models which cytoskeleton protein structure and function could be defective but this has not so far been formally demonstrated. PMID- 6283664 TI - [Angiomatous cutaneous lesions revealing arteriovenous fistulas (author's transl)]. AB - Reports of Bluefarb-Stewart syndrome have been infrequent. Two observations of this condition are described. Both patients are young men who presented with extensive angiomatous lesions of the feet, revealing distal arteriovenous fistulas. Surgical treatment was successful in one patient. Diagnostic criteria are discussed. As to pathogenesis the most likely hypothesis is that the arteriovenous fistulas are congenital. Bluefarb-Stewart syndrome can therefore be classified as one of the angiodysplasias. PMID- 6283665 TI - [Weber-Christian syndrome and chronic pancreatitis (author's transl)]. AB - The authors describe a case of cytosteatonecrosis associated with chronic pancreatitis. Clinical, biological (amylasemia and amylasuria) and radiological (arteriography) signs of pancreatic disease are noticed as well as important radiological lesions of the limbs. The cutaneous lesions are hypodermic nodules located on the lower limbs with swelling of the right big toe and a cutaneous abrasion. PMID- 6283666 TI - [Rheumatoid arthritis with monoclonal gammopathy. Retrospective review of twelve case-reports (author's transl)]. AB - Monoclonal gammopathy (MG) occurs in the course of rheumatoid arthritis (RA) in approximately 3.7% of cases. Twelve such cases were compared to two-hundred and fifteen controls with RA. RA was of later onset in patients with MG. Patients with MG had lower latex and Rose-Waaler titers; a decrease in these titres heralded onset of MG. The twelve patients with both RA and MG were then compared to nine-hundred and seventy-one controls with monoclonal gammopathy. As in previous studies, we recorded both IgM excess and unexplained lambda light chain excess. PMID- 6283668 TI - [About three cases of median nerve compression inside the carpal tunnel (author's transl)]. AB - The authors report about three cases of median nerve compression inside the carpal tunnel; they were caused by tendons or anomalous muscular bellies of the Palmaris Longus. The first two cases consider the anomalies of the distal insertion of the muscle, the third a volume anomaly; the first two findings allow to recall the hypothesis of a common philogenetic origin of the palmar aponeurosis and the Palmaris Longus. PMID- 6283667 TI - [Carpal tunnel syndrome due to mycobacterium bovis BCG (author's transl)]. AB - A 45-year-old woman, previously in good health developed chronic tenosynovitis with carpal tunnel syndrome a few weeks after wounding herself with a Pasteur pipette containing BCG cultures. She had received two injections of triamcinolone in the anterior annular ligament of the right carpa. The patient recovered after twelve months of antituberculous therapy associating isoniazid (300 mg/day), ethambutol (800 g/day), and rifampicine (400 mg/day). This is the first report of BCG tenosynovitis of the hand. The hazards of steroid injections into the tendon sheaths are pointed out. PMID- 6283669 TI - [Evaluation of treatment of myasthenia with thymectomy (author's transl)]. AB - The paper describes the effectiveness of treatment of myasthenia with thymectomy in 136 patients operated on, observed in hospital and 107 examined by questionnaire on late results after 10 years. Exclusive of myasthenia with thymoma, cures were obtained in 26.4%, marked improvement in 19.6%, moderate improvement in 21.4%, improved motor strength in 27.1%, and deterioration was observed in 3.8%. The assessment was based mainly on the amount of cholinergic drugs taken. Early mortality rate was 2,4%, and late mortality 1,9%. Relation of postoperative complications/respiratory and crisis of both kinds to mortality and intraoperative pleural injury also surgical approach and anatomic variants of the thymus were discussed. An influence of the number of proliferating centers on prognosis in myasthenia after thymectomy was nor demonstrated. Thymectomy in a generally accepted and most effective method of treatment in myasthenia. PMID- 6283670 TI - [Hepatic syphillis with amyloidosis and chronic diarrhea. A discussion of etiologic mechanisms (author's transl)]. AB - A sixty-four year-old woman was admitted for chronic diarrhea with severe weight loss. Investigations showed hepatomegaly, positive serologic tests for syphilis, and nephrotic syndrome with proteinuria. Anasarca occurred and the patient died shortly after admission. Necropsy showed sclero-gummatous hepatic syphilis, generalized amyloidosis and ulcerative colitis. These last two manifestations and their association with tertiary stage syphilis are discussed. PMID- 6283671 TI - [Congestive heart failure due to chronic alcoholism (author's transl)]. AB - In rare instances, chronic alcoholism leads to the congestive heart failure which is characteristic of alcoholic beriberi with edema. The affected patients is usually a young man with longstanding alcoholic intoxication and often with neurologic features. Onset is often sudden, with dyspnea on exertion, orthopnea, and palpitations. The clinical sings of cardiac failure are unequivocal. Roentgenography shows cardiomegaly mainly due to enlargement of the right cavities and of the pulmonary artery. ECG shows sinus tachycardia and abnormal repolarisation in the precordium. The cardiac output and the cardiac index are increased, as well as the coronary output. Pyruvic acid levels exceed 15 mg/l, the provoked hyperpyruvicemia test is abnormal, thiamine levels are low, and the tryptophane test is normal. The course is variable. Cardiac beriberi progresses by exacerbation and remissions. Prognosis is poor, with a risk of sudden death. However, with adequate treatment combining rest, a low sodium diet, alcohol withdrawal, diuretics, and vitamin B1 (IV) recovery occurs. Our two observations clearly fit this description. PMID- 6283672 TI - [Professional future after myocardial infarction (author's transl)]. AB - After a first myocardial infarction, the professional future of 86 patients is studied. 80 men and 6 women are included in the study. The mean age is 50.68 years. Manual workers prevail among affected patients, regardless of their age or of the site of the infarct. 78% of patients under 60 years resumed work, during the first threa months in more than half the cases, without any readjustments. The mean age of patients who resumed work is 47.6 years. Work was more often completely discontinued among office clerks and business managers than among other professions. Difficulties in finding readjusted jobs are greater for manual workers, especially if they belong to a small firm or if they are over 50. From a psychological and professional point of view, rehabilitation should be included in the management of myocardial infarction, even if it's impact on long-term mortality remains unknown. PMID- 6283673 TI - Detection of gastrointestinal bleeding with 99mTc-labeled red blood cells. AB - Using a modified in vivo Tc-99M red cell labeling technique, gastrointestinal bleeding scintigraphy was performed in 100 patients with GI bleeding. Sixty-two patients with melena or bright red blood per rectum had positive scintiscans. In comparison to results of angiography, endoscopy, surgery and contrast radiography, radionuclide scintigraphy correctly located the site of bleeding in 83% of patients. The procedures could be performed over a 24 hr period which increased the sensitivity of the test since 85% of the scintiscans were positive at one hr or greater after the onset of imaging. The procedure was more sensitive than angiography in detecting sources of GI bleeding. We conclude that GI bleeding scintigraphy 99mTc-red cells in an accurate and effective method to detect upper and lower GI bleeding in patients with acute intermittent gastrointestinal bleeding. PMID- 6283674 TI - Paraganglioma resembling pheochromocytoma. PMID- 6283675 TI - Adrenal defect in adrenomyelodystrophy. AB - Adrenomyelodystrophy (AMD), a variant of adrenoleukodystrophy, is associated with both neurologic and adrenal dysfunction. Data from an endocrinologic evaluation of a 41-year-old pigmented white man with AMD showed elevation in basal plasma ACTH, 11-deoxycortisol, 17-alpha OH-progesterone and progesterone concentrations, and low normal plasma cortisol levels. Free urinary cortisol and 17-ketosteroid secretion values were within normal limits. These data suggest that the principal adrenal enzymatic defect in this patient was a partial block of 11-hydroxylase and that the elevation of precursors was ACTH-dependent. However, this patient may have other enzymatic defects. PMID- 6283676 TI - Malignant chemodectoma of posterior mediastinum. PMID- 6283677 TI - Recurring cystosarcoma phyllodes associated with breast carcinoma. AB - A 59-year-old patient, who had had recurring giant fibroadenomas and cystosarcoma phyllodes over a 36-year period, presented with cystosarcoma phyllodes and adenocarcinoma of the breast. Only seven other well documented instances of the coexistence of these lesions have been reported. The prognosis and treatment of adenocarcinoma was not affected by coexisting cystosarcoma phyllodes. PMID- 6283678 TI - Thromboneutropenia and levamisole. PMID- 6283679 TI - An epidemic of acute conjunctivitis caused by enterovirus-70 in Singapore in 1980. AB - The clinical, epidemiological and virological findings of the 1980 epidemic of acute conjunctivitis were described. Enterovirus 70 was isolated from 7 (35%) of 20 eye swabs submitted for virus isolation, and the paired sera of eight patients showed a four-fold or greater rise in neutralising antibody titre to Enterovirus 70. The disease mainly affected children and young adults of all ethnic groups. Most of the cases contracted the infection at home. The mean secondary attack rate was 72.6% and the mean incubation period, four days. The clinical features were similar to the 1970 and 1975 epidemics caused by Coxsackievirus A24. Subconjunctival haemorrhage was observed in 10% of the cases. Most of the cases recovered spontaneously within a week. Transmission of infection within the home was either indirect, probably through fomites contaminated with eye or respiratory discharges of cases, or direct, through intimate person-to-person contact. During outbreaks, health education on simple personal hygiene should be highlighted. PMID- 6283680 TI - Herpes simplex virus in patients with aphthous ulcers in Bangkok. AB - Fifty-two paired sera and 40 single sera and swabs from the aphthous like ulcers of the tongue, gum, and buccal mucosa were collected from patients at a private clinic and from workers of canned fish packaging company in an attempt to isolate herpes simplex virus. Sixty-eight percent had positive HVH isolation with high ELISA antibody titers. Among 52 paired sera from patients, 68.4% had recurrent infections and 31.6% had primary infection. PMID- 6283681 TI - [Arduan - a new muscle relaxant]. PMID- 6283683 TI - Western diseases and their emergence related to diet. AB - Many of the commonest diseases in the economically more developed communities are characteristic of modern Western culture. Evidence is presented suggesting that they represent a failure of adaptation to the dramatic changes in diet that have been associated with the emergence of modern Western culture. Dietary changes aimed at the alleviation and prevention of these diseases are discussed and recommended. PMID- 6283682 TI - Systemic lupus erythematosus or mixed connective tissue disease? -- A case of 14 year old girl with viral myocarditis. PMID- 6283684 TI - Diverticular disease of the colon. The first of the Western diseases shown to be due to a deficiency of dietary fibre. AB - Diverticular disease of the colon is a new disease that appeared at the beginning of this century. It is now the commonest disease of the colon in the Western world, being found in 1 in 3 people of over 60 years of age. The pathogenesis of the disease involves excessive segmentation, but this does not explain its aetiology. The historical appearance of the disease on the clinical scene and its geographical distribution suggest that it is due to the removal of fibre from carbohydrates. The author treated 70 patients with symptomatic diverticular disease with a high-fibre diet. The results of this and the effects of bran are discussed. PMID- 6283685 TI - De novo malignant tumours in renal transplant recipients. AB - Malignant tumours developed in 8 of the 209 patients who received renal transplants at Groote Schuur Hospital, Cape Town, between October 1967 and December 1979. One of these patients developed disseminated carcinoma of the breast, a disease which had been present in the donor. He was therefore excluded from the study. At 3,4% the incidence of de novo malignant tumours in transplant recipients is half that previously reported in South Africa. No case of reticulum cell sarcoma or intracranial lymphoma was found. PMID- 6283686 TI - [International conference on mental deficiency in youth]. PMID- 6283687 TI - Left hepatic trisegmentectomy. PMID- 6283688 TI - Improved surgical resection of human brain tumors: Part I. A preliminary study. AB - Twenty-three patients underwent craniotomies for brain malignancies. One hundred eleven fluorescein-stained and 75 nonstained tissue biopsies were obtained for histopathological examination using a special arc-lighting system connected to a conventional surgical headlight. Of the 75 nonstained specimens, 71 (94.7%) were negative for tumor and 4 demonstrated the presence of tumor. Ninety-four (84.7%) of the fluorescent tissues obtained were reported as having neoplasia and the remaining 17 showed characteristics consistent with necrotic tissue debris. All stained tissues obtained in this study demonstrated visible fluorescence within neoplastic tissue margins harboring abnormally permeable blood-brain barriers. PMID- 6283689 TI - Experience with intraoperative CT scanning in brain tumors. PMID- 6283690 TI - HLA antigens in adults negative for antibody to Epstein-Barr virus (EBV). AB - Evidence for a genetic component in susceptibility to Epstein-Barr virus (EBV) infection was sought by comparing HLA A and B phenotypes of EBV antibody-negative Scottish medical students and randomly chosen antibody-positive controls. No statistically significant differences were observed, but three antigens, (A10, A29 and B15), were relatively underrepresented in the EBV seronegative group; findings which agree with data previously reported from a similar study in Los Angeles. A strong association between the HLA A1/Blank phenotype and EBV seronegativity, evident in the Los Angeles population, was not confirmed in the present study. PMID- 6283691 TI - Decreased effectiveness of chelation therapy with time after acute cadmium poisoning. PMID- 6283692 TI - Benzo(a)pyrene-induced early cytopathologic changes and peroxidase activity in the endometrium of ovariectomized rats. PMID- 6283693 TI - Dietary influences on theobromine-induced toxicity in rats. PMID- 6283694 TI - Coir fibre toxicity: in vivo and in vitro studies. AB - The biological activity of coir fibre, coir ash and their components were investigated in vitro by measuring the haemolytic activity and macrophage cytotoxicity. In vivo studies carried out by injecting guinea pigs intratracheally with coir fibres resulted in resolving granulomas. The observed haemolytic activity and macrophage cytotoxicity was more marked with coir ash compared with coir fibres. Chemical analysis of coir ash revealed the presence of toxic chemical constituents in appreciable amounts. PMID- 6283695 TI - Comparative cytotoxicity of DQ12-quartz and fly ash particles from coal combustion. AB - Fly ash samples obtained from different coal combustors were tested for cytotoxicity to alveolar macrophages in vitro by measuring the release of dehydrogenase into the culture media. Fly ash samples obtained from the combustion of a variety of coals in several types of combustors were not found to be cytotoxic when compared to a quartz sample of known toxicity. PMID- 6283696 TI - Distribution and retention of benzo(A)pyrene in rats after inhalation. AB - Tritium levels in tissues of rats were determined after inhalation of tritiated benzo(a)pyrene (3H-BaP; 500 micrograms/l; activity mass median diameter, 1-2 micrometers) for 1 h. Significant amounts of radioactivity were found in the nasal turbinates, trachea, larynx, lungs, tracheobronchial lymph nodes, kidneys, and liver immediately after exposure. Lower concentrations of radioactivity were found in the brain, testes, and spleen. Clearance of radioactivity from the respiratory tract occurred in two phases; a rapid phase where 50% of the radioactivity cleared by approx. 2 h, and a slower phase that continued for about 2 days after exposure. Radioactivity in the brain, testes, and spleen remained at about the same level during the first day after exposure and then decreased slightly during the second day. Benzo(a)pyrene inhaled in this particulate form was rapidly solubilized, translocated to internal organs, and excreted primarily in feces. PMID- 6283697 TI - [Hydrolases of the oral fluid in xerostomia]. PMID- 6283698 TI - [Glomus angioma of the maxillofacial area]. PMID- 6283699 TI - Pharmacokinetics of free catecholestrogens and catecholestrogen benzoates. AB - To provide a definite basis for studies on the biological effects of exogenously administered catecholestrogens, the time courses of the concentrations of these estrogens in serum, pituitary and CNS-tissues were studied in male rats after s.c. injection of either 150 microgram of 4-hydroxyestradiol or 2 hydroxyestradiol (dissolved in 200 microliter sesame oil/ethanol/ascorbic acid; 97.5/2.5/0.1; vol/vol/wt) or equimolar amounts of 4-hydroxyestradiol 3, 4 dibenzoate or 2-hydroxyestradiol 2,3-dibenzoate (dissolved in 200 microliter sesame oil). The injection of free catecholestrogens resulted in bolus-like elevations of the serum and tissue concentrations of the respective compound (max. values up to 9 ng/ml, half-life below 1 h) whereas the injection of catecholestrogen benzoates gave lower (max. values about 1 ng/ml) but prolonged elevations (half-life approx. 24 h and 32 h for 4-OHE2 and 2-OHE2) of the respective free catecholestrogen. PMID- 6283700 TI - [Effect of pH on ATPase activity and 18-O metabolism catalyzed by membrane preparations of Na,K-ATPase from guinea pig kidneys]. AB - The effect of pH in the incubation medium on ATPase activity and the direct 18O exchange, catalyzed by initial and treated by surface active agents membrane preparations of Na, K-ATPase has been studied. The pH dependence of the ratio between the direct 18O exchange and Na,K-ATPase activity was detected; this ratio being equal to 4,5 +/- +/- 0.6, 1.5 +/- 0.1 and 0.4 +/- 0.1 at the pH values of 6.1, 7.5 and 9.0, respectively. The pretreatment of preparations by DOC or histone H2a, used in both activatory and inhibitory-amounts, abolished the effect of acid pH and made the ratio practically the same at 6.1 and 7.5 pH values. Both agents did not affect the action of the alcaline pH value equal to 9.0. Triton X 100 was not able to change the pH dependence of the ratio. PMID- 6283702 TI - [Electromyography in the differential diagnosis of infant hypotonia]. PMID- 6283701 TI - Bluetongue virus precipitating antibodies in dairy animals in the Punjab. PMID- 6283703 TI - Prevention of platelet aggregation in patients with prostatic cancer during estrogen therapy. PMID- 6283704 TI - Enhanced tumorigenicity and hepatitis B virus genes in a human hepatoma cell line injected into immunosuppressed nude mice. PMID- 6283705 TI - Increased plasma epinephrine concentration due to its decreased conjugation in beta-hyperadrenergic essential hypertension. PMID- 6283706 TI - Stimulation of secretion of collagenase and prostaglandin E2 by synovial fibroblasts in response to crystals of monosodium urate monohydrate: a model for joint destruction in gout. PMID- 6283707 TI - Osteoclast precursors, mononuclear phagocytes, and bone resorption. PMID- 6283708 TI - 19-nor-deoxycorticosterone excretion in healthy and hypertensive subjects. PMID- 6283709 TI - Beta-adrenoreceptor down-regulation by dynamic exercise and upright posture in man. PMID- 6283710 TI - Skeletal muscle ionic composition and sodium transport activity in chronic alcoholism. PMID- 6283711 TI - Primary renal tumors in nonhuman primates. AB - In 17 nonhuman primates, nine females and eight males from 5 to 22 years old, there were 10 cases of renal carcinoma, four of renal adenoma, one nephroblastoma, one hamartoma and one transitional cell papillomatous hyperplasia. The most frequent clinical signs were anorexia, lethargy, weight loss, depression, and dehydration. Tumors were 0.1 to 10.0 cm in diameter. In neoplasms of tubular cell origin, papillary, tubular and solid histologic growth patterns occurred either separately or in combination. Thirty previously reported cases of primary renal tumors in nonhuman primates also were reviewed. PMID- 6283712 TI - Experimental vesicular exanthema of swine virus and San Miguel sea lion virus infection in phocid seals. PMID- 6283713 TI - Vesicular exanthema of swine and San Miguel sea lion virus: experimental and field studies in otarid seals, feeding trials in swine. AB - The naturally occurring disease caused by San Miguel sea lion virus in fur seals was characterized by small fluid-filled vesicles 1 to 25 mm in diameter on the nonhaired portions of the flippers. Early epithelial lesions contained multifocal sites of cell lysis. The resultant microvesicles enlarged and coalesced, forming grossly visible macrovesicles. Mature vesicles progressed to involve all layers of the epithelium but did not involve the underlying dermis. Intradermal inoculation of vesicular exanthema of swine virus type A48 or San Miguel sea lion virus type 2 into otarid (fur) seal pups caused plaque-like lesions around inoculated coronary bands. These swellings regressed without rupture by 96 hours postinoculation. One seal inoculated with San Miguel sea lion virus had a linear lingual erosion at ten days postinoculation. Virus was isolated from this site and from two uninoculated sites, the tonsil and testicle. Contact controls showed no evidence of infection. Virus was isolated in low titers from some sites of inoculation and draining lymph nodes from seals infected with vesicular exanthema of swine virus. Virus was recovered more easily, in higher titers, and from more tissues, from seals infected with San Miguel sea lion virus. Inoculated seals tested after four to ten days seroconverted. Feeding swine seal tissues from the inoculation experiments resulted in seroconversion in swine which were fed tissues from seals infected with vesicular exanthema of swine virus but not in those which were fed tissues from seals infected with San Miguel sea lion virus. PMID- 6283715 TI - Avian calicivirus. PMID- 6283714 TI - The pathogenesis of vesicular exanthema of swine virus and San Miguel sea lion virus in swine. AB - Vesicular exanthema of swine virus type A48 or San Miguel sea lion virus type 2, when inoculated intradermally into swine, resulted in fluid-filled vesicles at the sites of inoculation in the snout, coronary band, and tongue. Pigs that developed vesicles also had fevers. Secondary vesicle formation varied, depending on virus serotype. Viremia was found in one pig infected with San Miguel sea lion virus five days after infection. Virus was recovered from nasal-oral passages for up to five days after infection in both groups of pigs and from the gastrointestinal and urinary tracts of pigs infected with San Miguel sea lion virus. Neutralizing antibodies began to increase three days after inoculation and reached peak titers in seven to ten days. In the absence of secondary bacterial infection, healing was well advanced by ten days after inoculation. Lesions usually were limited to nonhaired portions of the integument and tongue. Individual epithelial cells became infected when a break in the skin allowed virus access to susceptible epithelial cells from either exogenous or endogenous sources. Individual infected cells ruptured and adjacent cells were infected, resulting in the formation of multiple microvesicles. Centrifugal coalescence of microvesicles led to formation of grossly visible macrovesicles. Lesions rarely developed from viral contamination of intact hair follicles. A mild virus-induced encephalitis was seen in pigs infected with vesicular exanthema of swine virus, and virus was recovered from brain tissue of pigs infected with San Miguel sea lion virus. PMID- 6283717 TI - The epidemiology of influenza as a zoonosis. PMID- 6283716 TI - Comparison of the efficacy of four inactivated infectious bursal disease oil emulsion vaccines. AB - A comparison was made between four inactivated infectious bursal disease (IBD) oil emulsion vaccines. Vaccine A, which was prepared from antigen derived from bursae of Fabricius, stimulated higher levels of IBD gel diffusion titres than vaccines B, C and D, which were prepared from antigens derived from embryo fluids. The progeny of parents vaccinated with vaccine A resisted IBD challenge for eight days longer than the progeny of any of the other vaccinated parents. PMID- 6283718 TI - [Metastases of nephroblastoma to bone in children]. PMID- 6283719 TI - IgG1 antibody in milk protects lambs against rotavirus diarrhoea. AB - Newborn gnotobiotic lambs were fed a diet of diluted evaporated milk supplemented either with normal ewes' milk or with milk obtained from ewes injected parenterally during gestation with rotavirus. Lambs fed 150 ml per day of milk collected 5 days after lambing from normal ewes were susceptible to rotavirus infection and diarrhoea, while lambs fed milk from vaccinated ewes collected either 5 or 12 days after lambing were protected. Analysis of the milk by column chromatography showed the anti-rotavirus activity to be in the fractions containing IgG1. PMID- 6283720 TI - Spontaneous rosette formation of pig and guinea pig thymocytes and peripheral blood lymphocytes: influence of proteases, oxidising and reducing agents, cytochalasin B and carbohydrates. AB - Spontaneous rosette formation (SRF) of pig and guinea pig thymocytes (Th) and thymus derived peripheral blood lymphocytes (PTL) was tested under different experimental conditions. Treatment with different proteases (trypsin, chymotrypsin, protease type VII) showed that SRF of pig Th and PTL is inhibited to a much higher extent than that of guinea pig Th and PTL. Cytochalasin B (CB) inhibited SRF of PTL of both species to a higher degree as compared to Th. Different carbohydrates neither influenced SRF of Th nor of PTL in both species indicating that simple carbohydrates are not the recognized component of SRF receptors. Treatment with oxidising and reducing agents did not influence SRF. PMID- 6283722 TI - Pigeon herpes encephalomyelitis: a review. PMID- 6283721 TI - Optimum conditions for the radioimmunoprecipitation assay for the major internal protein of bovine leukemia virus. AB - The sensitivity of the radioimmunoprecipitation assay for the major internal protein (p24) of bovine leukemia virus (BLV) was examined. Conditions were varied for the assay by 1) using larger amounts of serum and 2) increasing the amount of second antibody. Sensitivity for the p24 radioimmunoprecipitation test was greatest when 10 microliter of test serum and 200 microliter of rabbit anti bovine IgG were used. Under these conditions there were no discrepancies between the p24 radioimmunoprecipitation test and the radioimmunoprecipitation test for the surface glycoprotein (gp51) in 380 cattle from commercial dairy herds. PMID- 6283723 TI - [Use of the radial immunodiffusion test in the study of foot-and-mouth disease viruses]. AB - The radial immunodiffusion test (RIT) was used in the study of the quantitative interrelations between three subtypes of type A of the foot-and-mouth disease virus--A5, A10, and A22. A reverse proportional correlation was demonstrated between the size of the precipitation circles and the antibody concentration in the homologous sera, and a direct proportional correlation between the diameter of the circles and the amount of the antigen used. It was shown that one and the same antigen produced precipitation circles of a different size with several series of homologous hyperimmune serum, depending on the content of antibodies in these sera. It was found in experiments with early sera obtained on the 7th day of a foot-and-mouth disease infection in guinea pigs as well as in investigations with antiserum 140 S of the foot-and-mouth disease virus A5 that the early and 140 S sera used in RIT had higher specificity as against the hyperimmune sera. Stated is the necessity of checking the specificity of the early sera, which often reveals antibodies of the 7 S class. PMID- 6283724 TI - [Method of determining the biochemical behavior of clostridia]. AB - A fast and readily applicable method is suggested for the determination of the biochemical reactions of the spore producing anaerobic organisms. A pepton coloid medium with the addition of 0.15 per cent agar-agar was used as a basic nutrient medium. The cultivation of bacteria did not require any special conditions. Reading of the results was carried out with the addition of 0.1 per cent phenol red after 48 hours of incubation of the cultures. This method showed a number of advantages over the methods used so far. PMID- 6283725 TI - [Resistance of broilers to Marek's disease virus]. AB - Studied was the resistance, resp., susceptibility of the initial broiler lines - 66 and 77 of the Cornish breed, and 88 and 99 of the Plymouth Rhock breed, which are used in this country for the production of the four-line hybrid, - to the virus of the Marek's disease, isolate CT-1. A biologic assay was carried out with a total of 2080 young birds divided into three groups; I groups - infected abdominally; II group - contacts; and III group - controls. Certain breed, line, and sex differences were established in terms of resistance, resp., susceptibility of the investigated birds to the CT-1 isolate of the Marek's disease virus. More resistant were the birds of the Cornish breed as compared to those of the White Plymouth Rhock breed. A rising trend was observed in the resistance of cockerels in comparison with the poults of the respective line. Most susceptible to Marek's disease proved the birds of line 99, especially the poults that were currently used for the production of the mother parental form of the four-line hybrid. The birds with abdominal infection showed death cases as early as the second month of age, mortality rising up to end of the fourth month, while with the contacts and controls most death cases were observed at the fifth month of age. Essential decrease in mortality set in with all three groups after the birds reached six months of age. At a low rate of infections which was experimented with the contact and the control group there were not opportunities to fully discover the resistance of birds to Marek's disease. PMID- 6283726 TI - [Inhibiting activity of amantadine (1-aminoadamantane) on the transmissible gastroenteritis virus of swine in cell cultures]. AB - Studies were carried out on the inhibiting activity of amantadine - 1-amino adamantine (1-AA) on the virus of swine transmissive gastroenteritis, Rakovski strain, in cell cultures of pig kidney and thyroid gland. The inhibiting agent was tested in concentration of 6.25 to 100 microgram/cm3. Parallel titrations with and without 1-AA (25, 50, and 100 microgram/cm3) were used to establish the maximum difference in the titers 2 lg. The effect of inhibition was manifested at minimum concentration of 1-AA (12.5 microgram/cm3), and increased up to a concentration of 100 microgram/cm3. Within the interval 12.5-25.00-50.00 microgram/cm3 the doubling of the concentration of the inhibitor led to the decrease in the virus titer by 1 lg. In experiments to follow up the dynamics of the virus growth in the course of four days with and without 1-AA at the rate of 100 microgram/cm3 at varying multiplicity of infecting (from 0.5 up to 0.00005) the transient differences between the titers rose to 4.5 lg, while the differences between the peak values of the titer rose up to 2.5 lg. By means of seven consecutive passage of two variants of the virus selected by their decrease or increase in sensitivity to the inhibiting agent no change in their sensitivity to 1-AA was achieved. PMID- 6283728 TI - Equine arteritis virus-infected cells contain six polyadenylated virus-specific RNAs. PMID- 6283727 TI - [Virus neutralization microreaction to determine antibodies against the infectious bovine rhinotracheitis virus]. AB - Comparative studies were carried out with a total of 170 serum samples from cattle with the use of the virus-neutralization test and the virus-neutralization microreaction to demonstrate the presence of antibodies against the IBR virus. It was found that the two reactions were equally effective in terms of specificity. The coincidence of the results obtained with regard to the positive and negative tests was 97 per cent with variant alfa and 98 per cent with variant beta. Comparatively higher sensitivity was demonstrated with variant beta. It was established that the virus-neutralization microreaction was not influenced by the presence of antibodies against the virus in the initial cell cultures, it producing a well marked border line when reading the results. It was concluded that the virus-neutralization microreaction could in all cases be used instead of the virus-neutralization test as it is economical and is readily performed. PMID- 6283729 TI - Different histone families in intracellular SV40 nucleoprotein complexes with respect to the acetylation turnover. PMID- 6283730 TI - Novel localization of pp60src in Rous sarcoma virus-transformed rat and goat cells and in chicken cells transformed by viruses rescued from these mammalian cells. PMID- 6283732 TI - [Use of a technetium radionuclide complex to study blood-brain barrier permeability]. PMID- 6283731 TI - Covalent attachment of psoralen to a single site on vesicular stomatitis virus genome RNA blocks expression of viral genes. PMID- 6283733 TI - [Action by centimeter-range waves with subsequent nicotinic acid electrophoresis in the combined treatment of the sequelae of viral encephalitis]. PMID- 6283734 TI - [Ovarian and uterine tumors in rats after external exposure to different types of radiation]. PMID- 6283735 TI - [Problems of nephroblastoma in children]. AB - An evaluation of 388 cases of Wilms tumor within 13 years established a high proportion of advanced cases among children: stage II - 30.6; III - 35.4; IV - 27.6 and V (bilateral) - 6.4%. Main homeostatic indices remained unchanged until stages III-IV. Primary tumor and metastases can be detected by a complex of clinical, X-ray and radioisotope methods the effectiveness of which is determined by degree of tumor extension. Dissemination of tumor at different sites were detected by primary examination in 40.5% and at later stages in 15% of patients. Most metastases (96%) were detected within the first 12 months of therapy. They were significantly more frequent at stage III and their frequency was in correlation with the patient's age at the time of tumor detection. Out of 388 cases, 112 (28.9%) survived for more than 3 years, stage II - 57.1; stage III - 25.4 and stage IV - 4.6% included. 1.5 - 2 year-long courses of pre- and postoperative polychemotherapy proved to be effective, whereas in cases of stage III tumor they should be supplemented with irradiation of primary tumor, its bed, regional lymph collectors and metastases. These measures ensured a 3-year survival in 69.5% of stage II patients and in 32.3% of stage III patients. PMID- 6283736 TI - [Diagnosis and treatment of trophoblastic uterine tumors]. AB - An evaluation of 801 cases of different forms of uterine trophoblastic tumors treated at the Center in 1960-1981 is given. Chorionepithelioma was confirmed histologically in 488 cases. Modern means of diagnosis of uterine trophoblastic tumors are discussed. A retrospective analysis of treatment of patients is presented. The result of up-to-date combination chemotherapy of patients with uterine chorionepithelioma are reported. The role of surgery in the treatment of trophoblastic tumors is discussed. PMID- 6283737 TI - [Cholecalciferol metabolism of pregnant rats with vitamin D poisoning]. AB - Four and 24 hours after administering toxic doses of vitamin D3 the concentration of unchanged cholecalciferol in the blood plasma, liver, kidneys, osseous and muscle tissues in pregnant rats was lower than in the control animals. The content of 25-hydroxycholecalciferol in all test tissues of the pregnant rats was higher than in non-pregnant animals. Particularly great amounts of this metabolite are accumulated by the osseous tissue, kidneys and intestine, i.e. by vitamin D target tissues. During vitamin D poisoning the content of 25 hydroxycholecalciferol in embryos and placenta is fairly high, the content in the placenta being greater as compared to the test tissues of the pregnant rats. PMID- 6283738 TI - [Safety of grain grown on soils fertilized with carbonaceous rocks]. PMID- 6283739 TI - [Experiment in infecting the green monkey Cercopithecus aethiops with a leukocyte virus isolated in hepatitis A]. AB - The virus isolated in hepatitis A had been previously shown to passage in primary phytohemagglutinin-stimulated leukocyte cultures. The virus designated as leukocyte hepatitis virus (LHV) causes chromosome aberrations in the infected cultures. Some physico-chemical properties of LHV and its capacity for reproduction in other cell systems were studied. In serological surveys of patients and in studies of cellular immunity reactions of them LHV was found in cases of hepatitis. The possibility of producing a laboratory model of hepatitis in green monkeys, morphological reaction to inoculation with LHV, as well as the presence of specific antigen in the inoculated animals by immunofluorescence test using rabbit immune serum to LHV and FITC-labelled gamma globulins from hepatitis A convalescents were investigated. LHV antigen was found in the lymph nodes, spleen and liver. Active morphological reaction of lymph nodes and spleen was observed 30 days after inoculation of monkeys. The results indicate that in green monkeys LHV possesses marked reticulotropic properties without producing clinical hepatitis, therefore African grivet monkeys (Cercopithecus aethiops), apparently, cannot be used as experimental models for LHV studies in vivo. PMID- 6283740 TI - [Rapid identification of enteroviruses by countercurrent immunoelectrophoresis]. AB - The results of identification of 115 enterovirus strains by means of counter immunoelectrophoresis and the neutralization tests in cell culture are presented. Even at a high rate of agreement of the two methods, counter immuno electrophoresis was shown to be more rapid, economic, less labour-consuming but requiring concentrated viral antigens. PMID- 6283741 TI - [Use of a cocultivation method for detecting cytomegalovirus contamination of cell cultures of simian origin]. AB - At present, nonanthropoid primates are widely used as sources of cell cultures for manufacture of live viral vaccines. Simian cell cultures, particularly kidney cell cultures are also known to be frequently contaminated with cytomegaloviruses. The isolation of the latter is rather difficult due to the late appearance of the cytopathic effect in cell cultures of natural hosts. In the present study, the sensitivity of 4 methods virus isolation from the test cells was compared: the method of long-term cultivation of cells; the method of long-term cultivation with one subpassage of the cells; the method of cocultivation of the test cells by mixing with sensitive cells; and the method of co-cultivation by overlaying the test cells on an incomplete monolayer of sensitive cells. The latter method shortened the observation period and yielded a higher percentage of isolation of contaminating viruses from African green monkey kidney cell cultures. This method is supposed to be used in future for the detection of viral contamination of African group monkey kidney cell cultures utilized in manufacture of live viral vaccines. PMID- 6283742 TI - [Persistence of Japanese encephalitis virus clones in continuous HeLa cells]. AB - Some virological and biological properties of the H-N and H-P systems (cells of HeLa line clone persistently infected with the Nakayama and Peking I strains of Japanese encephalitis virus, JE, respectively) were studied. A feature distinguishing these systems from all previously described models of persistent infection (PI) of vertebrate cells with JE virus consisted in the lack of visible signs of virus-specific degeneration of the cultures. Attempts to free PI cells from persisting viruses by changing cultivation conditions or with virus-specific antibodies failed. The "zone" phenomenon frequently observed in PI as well as resistance of the cultures to superinfection with isologous viruses only appeared to indicate the presence of interfering particles in the H-N and H-P systems. PMID- 6283743 TI - [B-lymphocytes in chronic herpetic and adenovirus eye infections]. AB - Concentrations of serum immunoglobulins and the number of B-lymphocytes in the peripheral blood were determined in herpes and adenovirus infections of the eyes. In severe forms of ophthalmic herpes the level of IgM was found to be high at the peak of the disease. In ophthalmic adenoviral infection and level of IgA was decreased. At the peak of the disease, the relative and absolute number of B lymphocytes was decreased in severe forms of herpetic and adenovirus infections of the eyes. In the period between relapses the number of B-lymphocytes is completely restored. PMID- 6283745 TI - A glucagon-secreting alpha-cell carcinoma of the pancreas. PMID- 6283744 TI - Carcinoma of the lung in nonsmoking Chinese women. AB - The records of 452 lung cancer patients seen in San Francisco between 1972 and 1979 were retrospectively analyzed according to race, sex, findings at histology and smoking history. Of 31 Chinese women with adenocarcinoma or large cell undifferentiated carcinoma, 20 (64.5 percent) had never smoked, which is in sharp contrast to all other subsets. At least 12 of the women were of Cantonese origin. These data support the findings of previous studies carried out in Cantonese women residing in Hong Kong, Singapore and Hawaii. As the incidence of adenocarcinoma in nonsmokers is appreciably higher than that noted in Canton, the possibility must be considered that the cause may be the result of an interaction between some traditional and nontraditional exposures. PMID- 6283746 TI - The surgical management of primary carcinoma of the liver. PMID- 6283747 TI - Cytomegalovirus cecal erosion causing massive hemorrhage in a bone marrow transplant recipient. PMID- 6283748 TI - Dietary fibre, bacterial metabolism and toxicity of nitrate in the rat. AB - 1. A semi-synthetic diet, the semi-synthetic diet plus pectin, and a stock diet were fed to rats, and three metabolic functions of the caecal bacteria (reduction of amaranth, p-nitrobenzoic acid and nitrate) were measured in vitro. 2. No consistent differences were noted between diets for the reduction of amaranth and p-nitrobenzoic acid. 3. No consistent differences in nitrate reductase activity were noted for rats maintained on a stock diet or a fibre-free diet. However, the addition of 5% pectin to the latter diet resulted in a several-fold increase in nitrite production. 4. This increased nitrite production in vitro was associated with methaemoglobinaemia following the oral administration of nitrate to rats fed the diet containing pectin. Animals receiving the basal fibre-free diet were unaffected by nitrate. PMID- 6283749 TI - [Isolation of streptothricin resistant mutants from E coli K12, strain A19]. AB - Mutants with various levels of resistance to streptothricin were isolated from Escherichia coli K12, strain A19 after mutagenesis with N-methyl-N-nitro-N nitroso-guanidine and ethylmethane-sulfonate. Nourseothricin, a mixture of streptothricin F and D was the selection agent. Spontaneous resistant mutants could not be found. The streptothricin-resistant mutant E. coli A19 Stcr 2/2/1 shows cross-resistance to some of the aminoglycoside antibiotics investigated, but no cross-resistance to chloramphenicol and chlortetracyclin. These results indicate similar mechanisms of action of streptothricin and aminoglycoside antibiotics. PMID- 6283750 TI - [Relationship between nutrition and tumor formation]. AB - The tumours of man mainly develop under the influence of the long-term intake of carcinogens and cocarcinogens. There is a correlation between the level of the intake of meat and the development of tumours of the colon and the mamma. In vegetarians these forms of tumours very rarely appear. For their prevention it is necessary to decrease the consumption of sorts of meat and sausage rich in fat, so that less than 50 g of animal fat are taken in a day. It is to be recommended an increase of the consumption of food-stuffs rich in raw fibers. The caretenes, the vitamin A and the vitamin-A-acid proved as anticarcinogens. In smokers an intake of carotene and vitamin A which overwhelms the need is useful. With smoked food-stuffs in small quantities benzpyrene and nitrosamines are taken in, a restriction of the consumption of smoked meat and fish is not to be recommended. a carcinogenic effect have also aflatoxines, for which purpose no food-stuffs infested by fungi must be taken in. PMID- 6283751 TI - [Simultaneous occurrence of pituitary adenoma and thyrogenic hyperthyroidism]. AB - Subtotal tumour removal had been performed in a 34-year-old female patient for an extensive intra- and suprasellar expansive process. The considerably increased prolactin level did not decrease postoperatively, but normalised only after a three months bromocriptine treatment. The primary hyperthyroidism has been recovering after administering methimazolum. In a second case was reported on a 65-year-old female patient, suffering from rachitic dwarfism, stenosis of the aortic valve and tumour of the hypophysis, causing acromegaly, whose diabetes mellitus of contrainsular type could have been hardly balanced with insulin of a 128-unit-dose daily, and whose hyperthyroidism was due to an autonomous adenoma of the thyroid gland, first I-131 treatment was administered and she got into an euthyroid state. Six weeks following the removal of the acidophilic adenoma of the hypophysis administration of insulin could have been ceased, and the results of her growth hormone became normal. The clinical picture partly corresponds with Troell-Junet's syndrome. PMID- 6283752 TI - [Human papillomavirus and oncogenesis (author's transl)]. AB - Various types of human papillomavirus (HPV) are responsible for different warts. Characteristic of HPV infections is a lowered cell-mediated immunity (CMI), especially defective in cases of epidermodysplasia verruciformis induced by HPV5. The malignant transformation seems to be related with the type of HPV, and not with the grade of CMI defect and duration of the infection. A life-long infection with HPV3 in cases of EV does not induce a malignant transformation in spite of heavily depressed CMI. The oncogenic potential has seemingly HPV5 responsible for red plaques and pityriasis versicolor-like lesions characteristic of EV. Malignant transformation, although rare, may also occur in long-lasting cases of genital warts (Buschke-Loewenstein-disease) and juvenile larynx-papilloma. The most important problem is an early diagnosis and prophylaxis of genital cancers in women. This was made possible by cytological and virological studies of cervical dysplasia. In the condylomatous and non-condylomatous lesions of the colli uteri virus particles have been found using the immunoperoxidase technique and immunserum against disrupted viral particles, i.e. against the common HPV antigen. However, the type of HPV can not be determined by this method. Using the same immunsera virus particles were found also in the bowenoid genital papules. The best model for human oncogenesis remains EV. The clinical and pathological morphology, virological and immunological studies on the non-specific and HPV type specific humoral and cellular immunity are presented in detail. PMID- 6283753 TI - [Extent of beta-blocking action of S- and R-penbutolol during exercise testing (author's transl)]. AB - The effect of 40 mg penbutolol (pure S-form = laevo penbutolol) and 40 mg isopenbutolol (pure R-form) on heart rate and blood pressure during exercise testing was investigated under double-blind, randomized, placebo-controlled conditions in 9 probands. 1.5 and 5.5 hours after ingestion of 40 mg penbutolol, resting, standing, exercise, and recovery heart rate as well as systolic pressure during exercise displayed a significant decline. Diastolic blood pressure rose slightly 1.5 hours after penbutolol. Isopenbutolol had no significant effect on resting and standing heart rate. 1.5 hours after ingestion, a slight reduction in exercise and recovery heart rate could be confirmed. The decline, however, was significantly lower than that achieved with penbutolol. In comparison with its dextrorotatory isomer, isopenbutolol, penbutolol has an approximately 100-fold more potent effect on exercise heart rate. This is also reflected in the drug's duration of action. PMID- 6283755 TI - [Calcium and excitation-contraction coupling in smooth muscle cells]. PMID- 6283754 TI - CDC guidelines on sterilization of implants. PMID- 6283756 TI - [Common patterns in the nuclear organization of tumor cells]. PMID- 6283758 TI - [Tumor lipids and their effect on cell membrane structure and function]. PMID- 6283757 TI - [Biochemical mechanisms of action of a glucocorticoid-receptor complex on the expression of structural genes]. PMID- 6283760 TI - [Glucocorticoid receptors and their role in oncology]. PMID- 6283762 TI - [Local immunity in patients with herpes genitalis (clinical and electron microscope study)]. PMID- 6283761 TI - [Regulatory function of membranes during malignant growth]. PMID- 6283759 TI - [Polyamines in the evaluation of the effectiveness of cancer patient treatment]. PMID- 6283763 TI - [Comparative study of the effect of hormonally active and inactive forms of cholecystokinin octapeptide on the self-stimulation response in the rabbit]. PMID- 6283765 TI - [Intrathorakal nonchromaffin paragangliomas (chemodectomas) (author's transl)]. AB - Report of 3 benign nonchromaffin paragangliomas (chemodectomas) localised in the anterior mediastinum (female, aged 25), the posterior mediastinum (female, aged 55) and the right upper lobe of the lung (male, aged 40). These tumours arise from parasympathetic paraganglia. About 5-10% of the tumours are malignant. A survey of the literature about intrathoracic chemodectomas is given and age sex distribution, histology, morphological differential diagnosis, dignity and therapy of these tumours are discussed. PMID- 6283764 TI - [Contribution to the true non-African Burkitt's lymphoma (author's transl)]. AB - It is reported on a 6-year-old Middle European boy suffering from a nasopharynx tumor with involvement of a submandibular lymph node. The tumor was classified by morphologic, serologic and virologic data as a true or Epstein-Barr virus associated Burkitt's lymphoma, resp., which has been, up to now, seldom seen outside Africa. The patient died 3 months after diagnosis. Autopsy has shown tumor involvement of nasopharynx and soft tissues in the left neck region, several lymph nodes, kidneys, liver, spleen, and bone marrow. Cytologic, light and electron microscopic data are reported and the structural change of the tumor is referred to. PMID- 6283766 TI - [Gliomedullomyoblastoma. A contribution to the morphology and biological behavior of myogenic brain tumors (author's transl)]. AB - On the basis of an own observation and 38 cases found in the literature problems of histogenesis, classification and biological behavior of myogenic brain tumors and mixed neoplasms with myogenic elements are discussed. In a 2-year-old boy, 2 months after first clinical symptoms, a tumor of the walls of the 4th ventricle had been removed. The growth showed a large variety of histological structures. Irregularly arranged bundles of long muscle fibers with centrally located nuclei and cross striation were the most striking feature. Large cells with prominent nucleoli resembling ganglion cells were identified electronmicroscopically as rhabdomyblasts. Other areas were composed of astrocytic elements or of honeycomb like structures usually observed in oligodendrogliomas. In serial sections small cells with dark nuclei typical for medulloblastomas were found in small areas. The body died 15 months after the onset of symptoms. The autopsy revealed a relapse in midline structures of the cerebellum. The histological picture differed substantially from that of the primary neoplasm. It was composed only of small cells, showed a large number of collagenous fibers and corresponded to a demoblastic medulloblastoma. The histological variety is regarded as an expression of the pluripotency of neoplastic elements, which are evidently capable to form neuroectodermal as well as mesenchymal structures. It is emphasized that an overgrowth more anaplastic cells led to the described morphological changes. Gliomedullomyoblastoma is regarded as a special variant of medulloblastoma with an unfavourable prognosis. PMID- 6283767 TI - [Replication of the Aujeszky virus in vaccinated swine following experimental infection with large and medium amounts of virus]. PMID- 6283768 TI - [Comparative studies on susceptibility of viruses to ultraviolet rays]. PMID- 6283769 TI - Studies on bovine leucosis. IX. Excretion of bovine leucosis virus. PMID- 6283770 TI - [Effect of duration of the intervals between immunization with DPT vaccine on the serological indices in children]. AB - The influence of the interval between injections of adsorbed DPT vaccine on the effectiveness of immunization has been studied. The intensity of antibody formation to the pertussis component has been found to decrease 2.7-3.7 times if the interval between the injections constituting the course of primary immunization is prolonged for more than 6 months. The interval between the course of primary immunization and the first booster injection, which lasts 1.5-2 years in accordance with the currently accepted immunization schedule, is unreasonably long. Immunological characteristics in respect to pertussis have been found to decrease 8-45 times as early as 16 months after the first booster injection. Most of the vaccines catching pertussis at that time had the moderately severe form of this infection. PMID- 6283771 TI - [Mechanisms of in vivo suppressive effect of togaviridae and bunyaviridae on the activity of effectors of graft vs host reaction]. AB - Experiments on mice demonstrated the ability of 3 flaviviruses and 1 bunyavirus to suppress the activity of the effectors of the graft-versus-host (GVH) reaction. The conditions of the suppression of the primary immunological recognition were shown to differ in infections caused by different viruses. In experimental flavivirus infections caused by Langat, dengue 2 or yellow fever (strain 17D) viruses T-suppressor cells were activated, and their activity was realized only in respect to syngeneic or semisyngeneic target cells. In mice infected with Tahyna virus (a bunyavirus) no suppressor cells capable of suppressing the activity of the effectors of the GVH reaction were detected. The suppression of this reaction, not linked with the activity of the detected T suppressor cells, was observed in the Langat virus infection under conditions of bilateral incompatibility when both the donor and the recipient were infected. PMID- 6283772 TI - [Importance of a screening system in the prevention of diseases of the lumbosacral division of the peripheral nervous system]. AB - An automated screening system for prophylactic examinations is offered. The system enables one to reveal early compensated forms of lumbar osteochondrosis, as well as persons with an increased risk of that disease. The system is based on a statistical analysis of the lumbar osteochondrosis risk factors revealed in a particular individual, and involves the use of a computer. The practical use of this system at large industrial plants has made it possible to reveal potential patients to whom individual therapeutic and prophylactic complexes were administered. It is shown that the size of the risk groups in workshops depends on the character of the production operations performed and on the working conditions, the correlates with the prevalence of the disease in each particular collective body. PMID- 6283773 TI - [Use of stimulation electromyography in the diagnosis of peripheral nervous system lesions in herpes zoster patients]. PMID- 6283774 TI - [Differential evaluation of excitatory conduction rats along different segments of motor nerves and functional tests in the diagnosis of diseases of the human peripheral neuromotor apparatus]. AB - In 61 patients with neurono- and neuropathies and affections of the synaptic apparatus and the muscular matter of various etiology, the state of the peripheral nerves was examined. It was found that various forms of the pathology are characterized by specific changes in the speed of the stimulus transmission along various parts of motor nerves. The changes of the functional state can be seen especially clearly on comparing the speed of the stimulus transmission along the proximal and the distal parts of a peripheral nerve (the difference being named "transmission speed gradient"). In neuronopathies the transmission speed changes are prevailing in the proximal, in neuropathies in the distal, and in synaptic structure abnormalities in the extreme distal parts of the nerve. A 15 minute artificial ischemia of the nerve (a functional test) led to characteristic changes of the transmission speed, these changes correlating with the process stage. In cases of mild changes of the transmission speed the nerve was more sensitive, while in cases of pronounced changes more resistant to the ischemia. It was shown that the transmission speed changes in ischemia manifested themselves along the whole nerve, but not only in the site of its affection. In cases of the carpal canal syndrome the ischemia-induced lowering of the transmission speed in the nerve distal part was extremely sharp. PMID- 6283776 TI - Intralysosomal distribution of enzymes and components. PMID- 6283775 TI - [Surgical treatment of breast carcinoma - 30 years' experience]. AB - Although, carcinoma of the breast, because of its high frequency and many specific properties, belongs to the most studied of malignancies, it remains a fact that during the past 3 decades the incidence of mortality from this disease has changed little. The authors, drawing on their 30 years experience, demonstrate that the mortality rate and the survival rate depend on the kind of surgical intervention as well as the stage of development of the disease at the moment of operation. For this purpose a total of 655 operated cases were recorded (643 female and 12 male) using the Steinthal classification. In this series 68.1% were infertile; the upper outer quadrant was affected in 54% of cases; histologically the most frequent form was scirrhous adenocarcinoma. The greatest number of patients belonged to stage II and III, whereas the average time from the appearance of the first symptoms up to the medical examination was 6-12 months. Further spread of the disease and regional metastases were presented in more than 65% of cases. It was demonstrated that surgical procedure used depended on the extensiveness of the disease; modified radical mastectomy was performed in 54.2% of patients and Halsted's radical mastectomy in 40.15%. 185 patients were given the control examination. A strictly statistical analysis of the results confirmed that whereas the length of survival was not significantly affected by the kind of operation, the mortality rate did vary according to which surgical procedure was used. The author conclude that there is no best or unique surgical method and that each patient should be examined in a multidisciplinary way, and his treatment should be carefully planned. PMID- 6283777 TI - Effect of cold stress on catecholamines, cyclic AMP and cyclic GMP in hardened and unhardened men. PMID- 6283779 TI - Hyaline inclusions (Pseudopsammoma bodies) in meningiomas: immunocytochemical demonstration of epithel-like secretion of secretory component and immunoglobulins A and M. AB - Three meningotheliomatous meningiomas showed prominent formation of hyaline inclusions (pseudopsammoma bodies). Immunocytochemical examination by means of indirect immunofluorescence and PAP techniques demonstrated strong labeling of these inclusions by antisera to human secretory component (SC), immunoglobulin (Ig) A and IgM. A similar staining pattern was found in intracytoplasmic globules of a metastatic carcinoma. Hyaline structures of various types in atypical, psammomatous, and hemangiopericytic meningiomas were not stained with the antisera employed. By histochemical, immunocytochemical, and ultrastructural criteria, pseudopsammoma bodies in meningiomas are identical with intracytoplasmic inclusions of some epithelial neoplasms, especially of mammary and gastric carcinomas. Our study strongly corroborates Kepes' (1961, 1975) concept of secretory differentiation in meningiomas, and demonstrates an epithelial character at least of some meningiomas. PMID- 6283778 TI - Light- and electron-microscopic features of an endocrinologically active pituitary adenoma cultured in vitro. AB - A pituitary adenoma from an acromegalic female patient has been studied in cell cultures. Two populations of parenchymal cells, i.e., elongated shapes and big pale forms with irregular outlines, were distinguished light-microscopically. Ultrastructurally, three types of cells were found. Two of them contained secretory granules in varying numbers and sizes. Based on the proportion of the various cellular elements these cells were considered as counterparts of the elongated shapes being responsible for the GH secretion at different rates. The third cell type with very few secretory granules was identified with the big pale forms. Presumably, they could be exhausted GH-secreting cells. Besides fine structural characterisation, cultured cells of endocrinologically active human pituitary adenomas offer suitable models for studying some phases of exocytosis and membrane retrieval. PMID- 6283780 TI - Fine structure of inclusion body in the nucleus of Alzheimer glia type II in the brain of hepatocerebral degeneration. PMID- 6283781 TI - Estrogenic potency of oral replacement therapy estimated by the induction of pregnancy zone protein. AB - The induction of a highly estrogen-inducible plasma protein, the pregnancy zone protein/pregnancy associated alpha 2-globulin (PZP/PA alpha 2 G), was used to design a sensitive method to quantify and compare the potency of various estrogens commonly used in clinical practice. Serum PZP levels were followed in 100 post-menopausal women during 6 months' estrogen replacement therapy. Ethinyl estradiol caused more marked changes than the natural estrogens. Conjugated estrogens were more potent than estrone sulphate and estradiol-17 beta which had similar values. Estriol, irrespective of daily dosage, exerted a negative effect. The following sequence of equipotency was calculated: ethinyl estradiol = 100 x conjugated estrogens = 450 x estrone sulphate = 500 x estradiol-17 beta = 650 x estradiol valerate. These data were compared with other estimates of estrogenic potency from the literature. Compared with other clinical methods, PZP induction offers several advantages. Factors such as intestinal absorption, protein binding and intracellular metabolism are included in the net result of an increased protein synthesis which is easily followed in patients' sera during therapy. PMID- 6283782 TI - Low LH (hCG) receptor concentration in ovarian follicles in endometriosis. AB - The concentration of LH (hCG) receptors in the granulosa cells of ovarian follicles was assayed in 20 patients with histologically proven endometriosis and in 6 control patients. The levels of receptors in 15 patients with ovarian endometriosis (0.48 +/- 0.46 fmol/mg protein) and in 5 patients with endometriosis without ovarian involvement (0.51 +/- 0.52 fmol/mg protein) were lower (p less than 0.01 and p less than 0.05, respectively) than in control patients (1.32 less than 0.61 fmol/mg protein). The severity of the disease was not reflected in the results. The low receptor concentration in endometriosis suggests a failure in the mechanism mediating LH action in the follicle. PMID- 6283783 TI - [Malignant fibrous histiocytoma. Report of a case]. PMID- 6283784 TI - Pathogenesis of 1,2-dimethylhydrazine-induced carcinomas in rat intestine. I. The induction of mucin producing carcinomas in rat intestine. AB - Seventy four white male Wistar rats were divided into two groups and given weekly subcutaneous injections of 30 mg 1,2-dimethylhydrazine (DMH) 2HC1/kg body weight for 10 or 15 weeks, respectively, and the histological properties and distribution of DMH-induced rat carcinomas were investigated. The carcinomas induced by DMH were classified mainly into mucin producing carcinomas and non mucin producing ones. In the large intestine, the group treated for 15 weeks induced significant incidences of mucin producing carcinomas composed mainly of cells containing intracellular mucin. Mucin producing carcinomas consisted of poorly differentiated adenocarcinoma, signet ring cell carcinoma and mucinous carcinoma, and these often coexisted within the same tumor mass. Mucin producing carcinomas tended to develop in the proximal colon and caecum, while non-mucin producing carcinomas were frequent in the distal colon. This method was thought to be a useful model to study glycoproteins of cancer cells. PMID- 6283785 TI - Histopathological study of early lesions of pancreatoduodenal cancer. AB - The early lesions of pancreatoduodenal cancer, which was defined as those in which the site of origin is confined in a single certain anatomical part and the mode of early extension can be pursued, were found in 28 of 52 surgical specimens examined; 12 in the pancreatic portion of the common bile duct (PCBD), 12 in the common channel, 1 each in the terminal portion of the common bile duct and the terminal portion of the pancreatic duct, and 2 in the peripapillary zone of the duodenum. There were roughly two types of tumor growth; one annular flat-raised, histologically scirrhous tubular adenocarcinoma, principally found in PCBD and the other polypoid, histologically papillary or papillotubular adenocarcinoma, found everywhere. The staining properties of mucin was of no value in differential diagnosis for the site of origin. One typical carcinoid was observed in PCBD. The surface spread of carcinoma beyond the originating part was seen in 10 cases (36%), lymphatic invasion in 23 cases (82%), node metastasis in 9 cases (32%), and venous invasion in 6 cases (21%). Only 2 cases were mucosal carcinomas. From these figures it can be concluded that the carcinoma of this region is of high grade malignancy. PMID- 6283786 TI - [Central hypotensive effect of propranolol in relation to brain GABA receptors (author's transl)]. PMID- 6283787 TI - [Effects of gossypol acetate on neuromuscular transmission (author's transl)]. PMID- 6283788 TI - Effect of metrifonate on neuromuscular transmission. AB - The least detectable effect of acute cholinesterase inhibition (produced by edrophonium) in a normal subject is described. It consists of repetitive activity following the muscle response to a single nerve stimulus. Repetitive activity does not follow a second response at an interval of 30 msec. or 80 msec. There is no change in amplitude of the response to a single stimulus; nor to a second stimulus with the smallest dose of edrophonium. With higher doses of edrophonium the amplitude of the second response is reduced. 55 children were studied during treatment of urinary schistosomiasis with metrifonate (3 doses at 2 weekly intervals). No change in evoked muscle action potential amplitude attributable to treatment was detected. 3 children developed some repetitive activity 6 hours after their third dose, when erythrocyte cholinesterase inhibition was approximately 50%. PMID- 6283789 TI - Inhibition of enkephalin binding to opiate receptors by zinc ions: possible physiological importance in the brain. AB - Zinc ions can totally block stereospecific binding of 3H-met-enkephalinamide (2-D ala-5-L-methionine) to opiate receptor sites in synaptic membranes of the hippocampus, the cerebral cortex and the basal ganglia of the rat brain. Analysis of binding isotherms indicates that this inhibitory effect involves a decrease in both receptor affinity and the number of binding sites. Our data also suggest that the zinc ions react with essential SH-groups of the opiate receptor. The endogenous concentrations of zinc ions in the hippocampus, the cerebral cortex and the basal ganglia are compatible with the concentrations needed to inhibit opiate receptor binding in vitro. Especially the hippocampus contains a high concentration of zinc ion, which are localized exclusively in the giant boutons of the mossy fibers. Further, the hippocampal distribution of enkephalin and zinc ions is identical and confined to the mossy fiber zone. Hence, zinc ions may represent important modulators of opiate receptor binding in the central nervous system, particularly in the hippocampal mossy fiber zone. PMID- 6283790 TI - Clinical angiography with balloon catheter. PMID- 6283791 TI - Dosimetric intercomparison at the Scandinavian radiation therapy centres. I. Absorbed dose intercomparison. AB - A dosimetric intercomparison was carried out at all the centres in Denmark, Norway and Sweden. All the beam qualities (except conventional roentgen rays) used in radiation therapy were investigated. The ratio of the absorbed dose measured by us to that stated by the centre was determined. The mean ratio and standard deviation with the reference field-size were for 60Co gamma beams 1.001 +/- 0.014, for roentgen ray beams (4-45 MV) 1.017 +/- 0.023, electron beams with energy E0 less than 10 MeV 0.989 +/- 0.027, and electron beams with energy E0 greater than or equal to 10 MeV 0.996 +/- 0.034. The difference between the highest and lowest ratios was 17.8 per cent. A systematic difference due to the application of different protocols (NACP 1972 and 1980) was found. PMID- 6283792 TI - Diaphanography: mechanism responsible for the images. AB - Preliminary measurements showed that spectrophotometry of samples of tissue taken at operation from the female breast demonstrated no obvious correlation between transmission coefficient and pathology, except that thicker samples tended to transmit less light than thinner samples. However all the samples had absorption minima corresponding to the bands of oxyhaemoglobin. This simple fact led to the speculation that the density of blood in the tissues, (number of red cells per unit volume) was the factor giving rise to different colours of breast tissues and of breast pathology observed upon transillumination with white light in vivo. Examination by infrared colour photography and by spectrophotometry of transilluminated samples of blood diluted with normal saline confirms that the observed colour is dependent on red cell concentration. PMID- 6283793 TI - Proliferation kinetics and specific behaviour of G2 cells of the Bp8 mouse ascites sarcoma in vivo. AB - Growth characteristics of the Bp8 ascites sarcoma were investigated by means of evaluation of total cell number, the cellular DNA content, the mitotic index and the percentage of dead cells. Following intraperitoneal inoculation of 18 x 10(6) cells into NMRI mice a plateau level of about 1400 x 10(6) cells was reached after 7 days followed by a daily decline of 5 per cent up to day 14, the terminal stage of life. Based on the increase of the cell number and the distribution of cells in the cell cycle the flow rate through and the duration of the various parts of the cell cycle were calculated. Generally, the durations of all phases of the cell cycle including mitosis increased with age, most markedly after day 8 when the durations tended to become infinite. At the plateau phase the total number of G1, S-phase and mitotic cells decreased significantly while G2 cells remained at a constant level. This finding may be of interest in view of the concept of ascites tumour cells behaving as a regulated cell system. PMID- 6283794 TI - Dose response experiments using the mouse tail model. AB - Radiation induced tail necrosis in BALB/c mice was used to investigate the effects of tissue temperature and local oxygenation on the radiation response. Single doses of 250 kV roentgen-rays were given over the temperature range 32.5 to 37 degrees C and dose response curves obtained. The temperature dependence of the response was shown to be consistent with changing hypoxic status by the derivation of a simple model; this has practical implications for the treatment of patients using simultaneously combined radiation therapy and hyperthermia. PMID- 6283795 TI - Glottic carcinoma limited to the vocal cords. AB - Radiation therapy is generally considered to be the best primary treatment for early glottic carcinoma, with surgery reserved for local failure. In a series of 150 cases of T1 glottic carcinoma 138 were given radiation alone. In these patients the survival was 88 per cent at 3 years and 77 per cent at 5 years. Local control remained unchanged at 82 per cent after 6 years. When the results of surgery are included, local control was raised to 93 per cent. Preservation of the larynx was obtained in 92 per cent of survivors. Local extension or radiation dose levels had no influence on local control. The optimum dose could be lower than generally recommended. Voice quality after radiation therapy is superior to postsurgical laryngeal function. Since survival rates are identical, radiation therapy alone is the treatment of choice in all early glottic carcinomas. PMID- 6283796 TI - Carcinomas of the esophagus with synchronous or metachronous primary carcinoma in other organs. AB - Twenty-eight patients with esophageal carcinoma and one or more separate primary carcinomas in other organs were treated from 1960 to 1980, representing 8.3 per cent of the 339 cases of esophageal carcinoma observed during this period. The most frequent sites of the additional tumors were the oral cavity and pharynx (11 patients) and the stomach (12 patients). Awareness of the frequent combination of esophageal carcinoma with other primary neoplasms in the sites mentioned may lead to the detection of small curable malignant tumors. PMID- 6283797 TI - Tolerance of the central nervous system to photon irradiation: endocrine complications. AB - Dose-response isoeffect equations have been determined for hypothalamic pituitary insufficiency following cranial irradiation. Of particular importance is the occurrence of complications at doses substantially less than those commonly used for the treatment of central nervous system tumors. Such complications may be severe and potentially life threatening. These complications occur when a small midline 'target' volume containing the pituitary gland, infundibulum and adjacent inferior hypothalamic structures is irradiated. Direct pituitary irradiation is unlikely to be a factor, at least in some cases. The possible role of incidental hypothalamic irradiation in the control of acromegaly and pituitary dependent Cushing's syndrome is discussed. PMID- 6283798 TI - Modifications of small intestine lysosomal enzymes after irradiation at different times of the day. AB - The modification of lysosomal enzyme activities in animals irradiated with the same sublethal dose at 4 different times of the day is reported. The results confirmed the absence of circadian fluctuations in all the lysosomal enzymes and in protein content. A difference in behaviour between acid beta-galactosidase and beta-glucuronidase on the one hand and between acid phosphatase and cathepsin D on the other was evident in irradiated animals. The results showed that acid beta galactosidase and beta-glucuronidase increase from the early intervals after irradiation and reach the highest activity between 36 and 48 h. At these intervals autolysis phenomena, heavy cellular alterations and numerous phlogosis cells are present in the epithelium. Only beta-glucuronidase and acid beta galactosidase indicate the level of radiation injury. PMID- 6283799 TI - Effect of hyperthermia on a neurogenic rat cell line (BT4A) in vivo. AB - The effect of hyperthermia alone on the growth of the BT4A neurogenic tumour implanted into the feet of BD IX rats has been investigated. Following treatment by immersion of the tumour-bearing leg in a water bath at 42.0 to 45.0 degrees C a temporary retardation of tumour growth was observed but no cure. The lag phase before regrowth occurred was temperature and time dependent. A log-liner correlation was found between the surviving fraction previously found in vitro and the heat sensitivity of the cell line in vivo. PMID- 6283800 TI - Dietary fibre in type II diabetes. AB - Recent studies have indicated that diets rich in digestible carbohydrates and dietary fibre might be beneficial in the regulation of type II non insulin dependent diabetes (NIDD). Addition of the gel forming type of dietary fibre such as pectin and guar gum to meals or glucose solutions reduces post-prandial glucose and insulin response. Addition of cereal fibres in the form of bran seems to have long term beneficial effect improving glucose tolerance. Little is known, however, concerning effects of dietary fibre naturally occurring in food on postprandial glucose and hormone response. In the present study we prepared two breakfast meals which were similar regarding digestible carbohydrates but differed in their dietary fibre content. One of the meals, including whole grain bread and whole apples, contained 8.4 g of dietary fibre, and the other one, containing white bread and apple juice, 3.1 g. When given to eight NIDD, the fibre rich breakfast gave significantly lower blood glucose increment during the three hours following ingestion. The results indicate that foods rich in dietary fibre might be useful in the regulation of type II diabetes. PMID- 6283801 TI - Dietary supplementation of fibre (Lunelax) as a mean to reduce postprandial glucose in diabetics. AB - The postprandial glucose concentrations after a standardized breakfast of 12 type II diabetics were followed and the effect of supplementation of a fibre containing bulk-purgative (Lunelax) to the meal was investigated. It was found that addition of Lunelax reduced the mean increment glucose concentration with about 9%. The patients reported that Lunelax was convenient to take. PMID- 6283802 TI - [Use of the rabbit testicular cells in the plaque assay for herpesvirus hominis type 1 and 2]. PMID- 6283804 TI - Applications of hydroxyl proton chemical shifts of methyl substituted phenols as electronic and steric parameters in QSAR studies. PMID- 6283803 TI - Immune regulatory factors in sheep fetuses. AB - After an antigenic stimulus in vitro, a fraction was found in the supernatant fluids of spleen (S-1) and thymus (T-1) cell cultures of 70 to 80-day-old sheep fetuses infected with ovine adenovirus in utero. The fraction agreed with alpha-1 fetoprotein in mobility and its molecular weight was 67 000. Injected into adult sheep, S-1 and T-1 had a suppressive effect on the humoral immune response to both adenovirus and Clostridium perfringens D vaccine. Tests with peripheral lymphocytes suggested that fetal cell supernatants do not affect blastogenesis but markedly inhibit Ig synthesis as it could be demonstrated by immunofluorescence tests. PMID- 6283806 TI - Peripheral alpha-adrenergic activity in the affective disorders. PMID- 6283805 TI - [Mixed tumor of the skin. Apropos of 5 cases]. PMID- 6283807 TI - Double blind test with mianserin versus chlorimipramine. AB - The Authors report a double-blind study to compare mianserin and chlorimipramine efficacy in the treatment of two groups of 30 depressed patients. The trial confirms that mianserin has an antidepressant effect similar to that of chlorimipramine with fewer side effects. During the study some subjects were tested by Electrodermic Response to acoustic stimuli, comparing the results with the responses of patients treated with chlorimipramine. Furthermore, the plasmatic beta-endorphin level in 10 subjects treated with mianserin was compared with values found in basic conditions. The significant decrease of the plasmatic beta-endorphin level after treatment with mianserin suggests that the drug may act as an antidepressant, by varying the endorphinergic cerebral activity. PMID- 6283808 TI - Salbutamol treatment of depression. PMID- 6283809 TI - Neuroendocrine abnormalities in depressive illness. PMID- 6283810 TI - Menopause, depression, and plasma opioids. PMID- 6283812 TI - Covalent binding of metabolically activated hydrocarbons to specific microsomal proteins. PMID- 6283811 TI - Studies on the mechanism of stimulation of microsomal H2O2 formation and benzo(a)pyrene hydroxylation by substrates and flavone. AB - The addition of activators like flavone and hexobarbital to hepatic microsomes markedly stimulates H2O2 formation. The similar increase observed with flavone of microsomal hydroxylation of benzo(a)pyrene and its inhibition by catalase and methanol suggests but does not prove a necessary interaction of microsomal H2O2 production with benzo(a)pyrene hydroxylation. Hexobarbital and flavone-stimulated H2O2 formation is optimal at a stoichiometric relationship of these activators and NADPH. This implies either their direct participation as electron donors or their indirect involvement in electron transport by facilitation of stoichiometric substrate cytochrome P-450/NADPH flavoprotein interactions. Steady state kinetics data are consistent with a scheme in which the formation in microsomes of a complex of 1 mole of NADPH with NADPH-cytochrome P-450 reductase and 1 mole hexobarbital with cytochrome P-450 regulates H2O2 formation. PMID- 6283813 TI - Factors affecting the intracellular generation of free radicals from quinones. AB - Isolated hepatocytes do not liberate appreciable amounts of superoxide into the external medium. Simple quinones stimulate the release of superoxide up to 15 nmol/min/10(6) hepatocytes. Superoxide release stimulated by a variety of simple quinones and more complex antitumor quinones was maximal at a quinone one electron reduction potential of -70 mV. This was qualitatively similar to the pattern of superoxide formation seen with NADH-cytochrome b5 reductase and NADH: ubiquinone oxidoreductase. Superoxide production by NADPH-cytochrome P-450 reductase was maximal at a quinone single-electron reduction potential at -200 mV. Phenobarbital pretreatment had no effect on superoxide formation by hepatocytes suggesting that NADPH-cytochrome P-450 reductase activity is not rate limiting for quinone stimulated superoxide formation. Sulfonated stilbenes, specific inhibitors of anion exchange, had no effect on the release of superoxide by hepatocytes suggesting that superoxide is not transported through anion channels in the plasma membrane. Pretreatment of hepatocytes with 10(-5) M diethyldithiocarbamate produced over a two fold increase in the release of superoxide. PMID- 6283814 TI - Roles of UDP-glucuronosyltransferase in the inactivation of benzo(a)pyrene. PMID- 6283815 TI - Metabolism of benzo(a)pyrene and benzo(a)pyrene 4,5-oxide in rabbit lung. AB - For several years our laboratory has been investigating the biotransformation of various environmental pollutants by lung. Studies have been performed with pulmonary subcellular fractions, purified monooxygenase and glutathione transferase enzymes, and preparations having intact cellular structure including the isolated perfused lung and cell fractions enriched in alveolar macrophages, Clara cells and alveolar type II cells. Collectively, these investigations have identified several metabolic factors which may contribute to the pulmonary toxicity mediated by certain polycyclic aromatic hydrocarbons (PAH). First, although lung has low overall cytochrome P-450-dependent monooxygenase activity for many substrates, relative to liver, this activity is localized in only a few cell types and specific activity in certain cell types, such as the non-ciliated bronchiolar epithelial (Clara) cell, can be high. Second, oxidative metabolites of benzo(a)pyrene tend to accumulate in pulmonary tissue due, at least in part, to the low ability of lung (relative to liver) to conjugate and detoxify phenolic, dihydrodiol and epoxide metabolites. Thus, products such as benzo(a)pyrene 7,8-dihydrodiol are available for further cytochrome P-450 dependent oxidation to ultimate carcinogens and cytotoxins. Moreover, the lung is efficient in removing benzo(a)pyrene 4,5-oxide and presumably other oxidized PAH metabolites, from the bloodstream. Consequently, the uptake of relatively stable electrophilic metabolites released by the liver may also contribute to pulmonary toxicity. PMID- 6283816 TI - Modulation of epoxide hydrolase activity; effect on the activation of benzo[a] pyrene and its covalent binding to DNA in the nucleus. PMID- 6283817 TI - Benzo(a)pyrene metabolism and enterohepatic circulation in the rat. PMID- 6283819 TI - Oxidative metabolism of the synthetic estrogens hexestrol and dienestrol indicates reactive intermediates. PMID- 6283818 TI - Effects of methanol and chloramphenicol on CCl4 toxicity. PMID- 6283821 TI - A novel pathway of nitrosamine metabolism in liver microsomes: denitrosation of nitrosamines by cytochrome P-450. PMID- 6283820 TI - The effect of substrates and inhibitors of monoamine oxidase on hepatic dimethylnitrosamine metabolism and mutagenicity. PMID- 6283822 TI - Molecular mechanism of the bactericidal action of myeloperoxidase-H2O2-chloride. PMID- 6283823 TI - Release of superoxide anion and enhanced candidacidal activity as a manifestation of macrophage activation: studies with muramyl dipeptide. PMID- 6283824 TI - Modulation of the inflammatory response by the neutrophil myeloperoxidase system. PMID- 6283825 TI - Oxidative damage to lysosomal enzymes in human phagocytosing neutrophils. AB - During phagocytosis neutrophils from 8 patients with chronic granulomatous disease released 2-3 times more activity of lysozyme and beta-glucuronidase than did normal neutrophils. This difference was caused by the partial inactivation of these enzymes by normal neutrophils. The inactivation of granule enzymes depends on oxidative products and takes place mainly in the phagolysosomes. Myeloperoxidase is involved in this phenomenon. PMID- 6283826 TI - The secretion of lysosomal enzymes by human polymorphonuclear leucocytes (PMN) and its modulation by serum complement. PMID- 6283828 TI - Discrepancies in the oxygen balance of whole human neutrophils and neutrophil homogenates. AB - The oxygen consumption, superoxide production and hydrogen peroxide generation was studied in human neutrophils phagocytosing zymosan particles. Application of sodium azide, as an inhibitor of catalase, and/or 1,3-bis(chloroethyl)-1 nitrosourea (BCNU), as an inhibitor of glutathione reductase, led to the conclusion that neutrophils convert about half of the oxygen consumed in the respiratory burst to hydrogen peroxide; the other half is used for formation of organic peroxides, disulfide bridges, etc. These products are rapidly degraded to water by catalase and/or the glutathione redox cycle. Reduction of exogenous cytochrome C accounted for only about 15% of the consumed oxygen. Neutrophil homogenates contain a badly damaged oxidase system, because oxygen consumption and hydrogen peroxide formation were only about one-tenth of that observed with whole cells. In contrast, cytochrome-C reduction was about three times as high as that found with intact cells. Probably, cytochrome C partly reconstitutes damaged oxidase systems, thus artificially increasing the oxidase activity. We conclude that cytochrome-C reduction is not a good parameter to characterize cell-free oxidase preparations. PMID- 6283827 TI - The respiratory burst of phagocytic cells: facts and problems. AB - 1. The so called "soluble" oxidase(s) are not involved in the respiratory burst of guinea pig and human granulocytes and of guinea pig peritoneal resident and elicited macrophages. 2. The activation of the oxidation of NADPH by a membrane bound NAD(P)H oxidase is the main mechanism responsible for the activation of the respiration of phagocytes. 3. The oxidase is inactive in resting cells and the activated form works on the plasma membrane. 4. More than one mechanism is operative in the oxidation of NAD(P)H by cell free particles in vitro. These mechanisms vary in relation to the conditions of assay (pH and concentration of substrate). 5. Under optimal conditions in vitro the enzymatic oxidation of NADPH practically involves the univalent pathway of oxygen reduction with stoichiometry of two nanomoles of O2 formed for one nanomole of NADPH oxidized. 6. Also in intact cells all O2 is first univalently reduced to O2 and then discharged outside the cell or in the phagocytic vacuoles. 7. The main reactions involved in the O2 balance in intact cells are the univalent reduction of O2, the dismutation of O2 to H2O2 and the degradation of the peroxide through catalatic and peroxidatic mechanisms. 8. The total oxygen univalently reduced by the activated oxidase is 2-4 folds the net oxygen consumed by the cells, depending on the mechanism of H2O2 degradation. 9. All the rate of extrarespiration is accounted for by the rate of oxidation of physiological concentration of NADPH by the membrane-bound enzyme. This adequacy can be observed only under appropriate experimental conditions, because the high activity of the oxidase is not a permanent state. PMID- 6283829 TI - Apparent Km of leukocyte O2 and H2O2 forming enzyme for oxygen. PMID- 6283830 TI - Formation of superoxide anions and hydrogen peroxide by polymorphonuclear leukocytes stimulated with cytochalasin. PMID- 6283833 TI - A short transient increase in cyclic adenosine 3', 5'-monophosphate levels of neutrophil granulocytes following exposure to chemotactic factors. AB - Stimuli which are chemotactic for the retrospective neutrophils always produce a transient increase in cAMP within the first minute whereas the chemokinetic or other non-chemotactic stimuli tested have no such effect. The question as to whether the transient increase of cAMP levels plays any direct role in the transduction of a chemotactic stimulus cannot be answered. Restimulation experiments indicate that the extent of the cAMP response is not a reliable indicator for the chemotactic responsiveness of neutrophils; there is no quantitative correlation between these two parameters. Further detailed kinetic and quantitative studies of the cAMP system would be needed to clarify this point. PMID- 6283832 TI - The respiratory burst in human polymorphonuclear leucocytes stimulated by particles. PMID- 6283831 TI - Hydrogen peroxide production in a cell-free system: evidence for the involvement of a chain reaction. PMID- 6283834 TI - Chemiluminescence and the study of phagocyte redox metabolism. PMID- 6283835 TI - An alternative mechanism for the production of hydroxyl radicals by stimulated neutrophils. PMID- 6283836 TI - Role of serine proteases in superoxide production by human neutrophils, monocytes and basophils. PMID- 6283837 TI - Release of the membrane-calcium and its relation to the superoxide formation by polymorphonuclear leukocytes. AB - The relationship between the intracellular translocation of calcium from the storage pool and the oxidative metabolism was studied. An intracellular calcium antagonist, TMB-8, inhibited the release of superoxide induced by a calcium ionophore A23187 and the inhibition was relieved by the addition of calcium ions. The release induced by cytochalasin D or by the ingestion of bacteria was similarly inhibited by TMB-8. The mobilization of intracellular divalent cations of leukocytes was monitored by a fluorescent probe, CTC. When the CTC-loaded cells were stimulated with cytochalasin D or E. coli, a fluorescence change ascribable to the release of calcium from the intracellular hydrophobic environment was observed. The dose-response curve of the fluorescence change and that of the superoxide release of th cells were very similar. TMB-8 inhibited both metabolic and fluorescence changes in parallel. The results support the hypothesis that an intracellular translocation of calcium is stimulated the oxidative metabolism of leukocytes. PMID- 6283838 TI - Peroxidatic activity distinct from myeloperoxidase in human monocytes cultured in vitro and in alveolar macrophages. AB - Human monocytes develop a peroxidatic activity (PA) in rough endoplasmic reticulum (RER) after adherence or after culture in semi-solid medium. This enzyme activity disappears after three days of culture in the majority of macrophages derived from adult monocytes but persists for one week in macrophages derived from neonatal monocytes. The PA is due to an enzyme distinct from myeloperoxidase (MPO), since monocytes from a patient with MPO deficiency develop the same PA as that of normal monocytes after adherence. By its localization and other characteristics, PA of adherent monocytes resembles that of rodent macrophages. We therefore investigated whether human alveolar macrophages exhibit PA, using a sensitive cytochemical method which prevents inhibition by aldehyde in adherent monocytes. In various pathological cases, four types of macrophages could be identified: the majority were peroxidase-negative, a small percentage was of exudate type exhibiting a PA in granules as blood monocytes, while few macrophages were intermediate, possessing only PA in RER i.e. of type resident and a smaller proportion had PA in RER and in granules i.e. exudate-resident macrophages. These findings demonstrate that human macrophages and adherent monocytes may exhibit PA in RER as has been reported for rodent macrophages. The true nature and function of the enzyme responsible for this PA, which is distinct from MPO, remains unknown, but some arguments seem to suggest its role in prostaglandin synthesis. PMID- 6283839 TI - Functional and metabolic abnormalities of diabetic monocytes. PMID- 6283840 TI - Round table on the clinical application of leucocyte function tests. PMID- 6283841 TI - Actions of leukotrienes on vascular, airway, and gastrointestinal smooth muscle. PMID- 6283843 TI - Comparative biological activity of natural and synthetic leukotrienes. PMID- 6283842 TI - Pulmonary and vascular actions of leukotrienes. PMID- 6283844 TI - Identification of leukotrienes in the sputum of patients with cystic fibrosis. AB - Sputum extracts from 16 of 25 adult patients with CF produced a slow, sustained contraction of isolated guinea pig ileum which was partially inhibitable by the SRS-A antagonist FPL 55712. Following progressive purification and HPLC, the major ileum-contracting substance was identified as LTD4. LTC4 was also present but in smaller amounts. The sputum contained other SRS-A-like activity which was not inhibited by FPL 55712; this substance was of a smaller molecular size than LTD4. The presence of LTB4 was also established using HPLC and an assay of neutrophil chemotaxis. These studies raise the possibility that leukotrienes and other lipoxygenase products may contribute to the progressive lung disease characteristics of CF. PMID- 6283845 TI - Stimulus-secretion coupling in the human neutrophil: the role of phosphatidic acid and oxidized fatty acids in the translocation of calcium. AB - Neutrophils, stimulated via their surface receptors by ligands such as f-Met-Leu Phe or immune complexes, release membrane calcium from intracellular stores and accumulate calcium from the extracellular medium. Both mechanisms provide for a rise in the intracellular concentration of free calcium which appears to be required for later responses of the cell: release of lysosomal enzymes and the generation of superoxide anion as neutrophils undergo an aggregation response. Exogenous ionophores, such as A23187 and PGBx, which move calcium bidirectionally across lipid bilayers, bypass the ligand-receptor step, thereby providing the necessary rise in intracellular calcium. In the course of the remodeling of membrane lipids--within 5 sec after their exposure to a stimulus--neutrophils generate phosphatidic acid, at least in part at the expense of phosphatidyl inositol. Because phosphatidic acid is the only phospholipid whose action in model membranes mimics the action of exogenous ionophores (as it is formed early enough and in sufficient quantities to account for uptake of extracellular calcium), we propose phosphatidic acid as a promising candidate for the role of endogenous ionophore. This ionophoretic action may serve to amplify signals launched by primary changes at the plasma membrane from whence calcium is mobilized during stimulus-secretion coupling. The release of oxidation products of arachidonic acid, be it via the cyclo-oxygenase or lipoxygenase pathway, is a concomitant of neutrophil activation. Indeed, the pathways by which oxidative products of arachidonate are formed may also be influenced by stimulus-specific changes in membrane phospholipids, not the least interesting of which is the generation of phosphatidic acid. PMID- 6283846 TI - Mediation of leukocyte components of inflammatory reactions by lipoxygenase products of arachidonic acid. AB - The leukocyte lipoxygenase products LTB4 and 5-HETE elicit human neutrophil and eosinophil chemotactic responses in vitro and in vivo (Fig. 4), and are present at elevated concentrations in the lesions of some human inflammatory diseases, such as rheumatoid arthritis and the spondyloarthritides. The chemotactic potency of LTB4 is similar to that of the minor fragment of the fifth component of human complement, termed C5a, and is 30- to 300-fold greater than that of 5-HETE and of other natural DHETE isomers. Human neutrophils possess distinct subsets of chemotactic factor receptors that are specific for LTB4 and for 5-HETE, as demonstrated by the selective competitive inhibition of the chemotactic responses to the parent stimuli by acetyl LTB4 and 5-HETE methyl ester, respectively, and by the failure of the lipid chemotactic factors to bind to isolated membrane protein constituents of the human neutrophil receptors for chemotactic formyl methionyl peptides. LTB4 and 5-HETE also elicit human neutrophil and eosinophil chemokinesis, stimulate the uptake of calcium and D-glucose, and enhance the expression of C3b receptors on the leukocytes; however, they exert only a minimal effect on superoxide generation and lysosomal enzyme release. LTB4, but not 5 HETE, stimulates the release of calcium from previously unexchangeable intraneutrophil pools, as has been described for potent peptide chemotactic factors. Although far less potent than LTC4 and LTD4, LTB4 constricts peripheral airways, enhances mucous secretion in the airways of the lung, and dilates and enhances the permeability of the microvasculature in skin and other organs (Fig. 4). A variety of leukocyte functions, including chemotaxis, D-glucose uptake, and lysosomal enzyme release, are impaired in association with the depletion of endogenous lipoxygenase products. Exogenous 5-HETE reverses some of the functional deficits of HETE-depleted leukocytes. Inhibition of leukocyte lipoxygenase activity also suppresses the intracellular content of hydroperoxyeicosatetraenoic acids and of novel polar metabolites of arachidonic acid that may be critical to the activation of human neutrophils and eosinophils. Thus LTB4 and the less potent 5-HETE are active extracellular mediators of the leukocytic components of hypersensitivity and inflammation and may also serve an important role as intracellular mediators of leukocyte function. PMID- 6283847 TI - Biological activities of leukotriene B4 (isomer III). PMID- 6283848 TI - Novel leukotrienes and lipoxygenase products from arachidonic acid. PMID- 6283849 TI - Effects of leukotrienes on in vitro neutrophil functions. PMID- 6283851 TI - Double dioxygenation of arachidonic acid in leukocytes by lipoxygenases. PMID- 6283850 TI - Effect of lipoxygenase products on leukocyte accumulation in the rabbit eye. PMID- 6283852 TI - Visna virus meningoencephalomyelitis in goats. PMID- 6283853 TI - Pathomorphology of the intestinal mucosa in diarrheic calves. PMID- 6283854 TI - Hog cholera antibodies in pigs vaccinated with an Aujeszky-vaccine based on antigen produced in IB-RS-2 cells. PMID- 6283855 TI - Enhancement of the biological activities of deoxyribonucleic acid extracted from canine venereal tumours. AB - Deoxyribonucleic acid (DNA) extracted from canine venereal tumours was capable of changing normal cells to morphologically abnormal cells in an in vitro system of L-fibroblasts. The observed effects were proportional to the concentration of the nucleic acid extract used; the higher the concentration, the more pronounced the effect. Inactivation of this effect by deoxyribonuclease indicated that the active nucleic acid moiety was DNA. Purified RNA did not appear to contribute to the observed activity. The activity was enhanced by diethyl-amino ethyl-dextran (DEAE-D) and dimethyl sulphoxide (DMSO) and the data showed that 20% DMSO was superior to 300 g/ml DEAE-D as an enhancing agent. PMID- 6283856 TI - Reflection effects of vaso-active drugs on arterial pulse propagation velocities measured in-vivo in a dog. AB - Blood-flow velocity signals have been recorded simultaneously at the proximal and distal sites of four arterial segments in an anaesthetized dog under two vaso active conditions (vaso-dilation and vaso-constriction) using a continuous wave Doppler-shift ultrasonic technique. Spectral analysis produced sonograms of these signals. Phase delays between flow pulses in each segment were obtained from the sonograms and the variation of the resulting phase velocities with frequency has been discussed. Foot-to-foot transit time of the sonogram waveforms for each segment also gave an apparent pulse wave velocity, PWVff (Pulse wave velocity foot-to-foot) and this has been compared with the corresponding phase velocity of the higher harmonics, PWVhh in determining the true or characteristic pulse propagation velocity in the arterial pathways. PMID- 6283858 TI - Nigerian university students' characteristics as measured by Eysenck personality questionnaire. AB - Eight hundred and seventy male and female freshman university students completed the Eysenck Personality Questionnaire (EPQ) as part of a study of the social, personality and mental health characteristics of Nigerian university students. The findings show that the EPQ is, in general, applicable in Nigeria, but there is need for further research to clarify differences between Nigerian and British subjects. PMID- 6283857 TI - Conjugated hyperbilirubinaemia in Nigerian infants. AB - A prospective study of 101 Nigerian infants with conjugated hyperbilirubinaemia seen over 6 years shows that extrahepatic biliary tract obstruction, idiopathic hepatitis, and bacterial infections were the common causes. A firm diagnosis was based on clinical, biochemical and histological features when the patient presented early. However, most of the infants presented late and the superimposed features of prolonged cholestasis made differentiation of the probable causes difficult. Erythrocyte peroxide haemolysis test and laparatomy aided diagnosis in these cases. Seventy five per cent of the patients with sepsis treated with antibiotics, and 70% of those with hepatitis treated symptomatically, recovered. Surgery was successful in only 15% of the patients with biliary tract obstruction. These were those who had either diverticulum of the common bile duct, localized atresia or stenosis or in whom biliary obstruction was due to viscid bile. PMID- 6283859 TI - Tracheal-tube size requirement of adult Nigerians. AB - A clinical study of the tube size (diameters) requirement of Nigerian adults (with no airway pathology) of both sexes was made. A prospective study showed that most (98.87%) Nigerian adult female patients require 9.0mm, while most (82%) male patients require 10.0mm tracheal tubes. Each of these sizes, being one size larger than that previously used, should reduce the work of breathing further as shown from an experimental study and is suggested for routine use in this environment. PMID- 6283860 TI - Causes of mortality in an African city. AB - Registered deaths from the vital statistics registration system of Lagos City, a system that was judged to be 60% complete, were analysed for the year 1977. Nearly 40% of total registered deaths were from infections, parasitic diseases and motor vehicle accidents. Of the reported deaths, 17.3% were from ill-defined conditions. Deaths from neoplasms, diseases of the nervous and sense organs, diseases of the digestive and genitourinary systems as well as those from congenital anomalies are relatively less frequent. Maternal mortality appears to be very high. The age pattern of mortality is different from that in the developed countries, a high proportion of the deaths in Lagos City being those of children aged under 5 years. Infant mortality is dominated by perinatal causes which constituted a huge 38.4% of deaths of infants under 1 year, the other important causes being dysentery and diarrhoea, pneumonia and tetanus. Among adults, death from motor vehicles accidents is the most important cause accounting for more than 26% of deaths in the age group 15 years and above. Other important causes of adult deaths are cerebrovascular disease, hypertensive disease, heart disease, pneumonia, dysentery and diarrhoea and complications of pregnancy. Well organized health services stressing antenatal care, preventive and health education services are needed to effect a reduction in mortality and bring about a general improvement in the health of the people. PMID- 6283861 TI - Teratomas in adult Nigerians. AB - Over a six-year period, 132 teratomas were seen at the Ahmadu Bello University Hospital in Zaria, Nigeria. Twenty-seven of these, comprising most of the extragonadal tumours were seen in infants and children. Of the 105 tumours found in adults, ninety-four were benign cystic teratomas of the ovary occurring mostly in the third and fourth decades of life. There were three instances of malignant transformation of a previously benign cystic ovarian teratoma and four cases of struma ovarii, one of which was admixed with trabecular carcinoid but none of which was associated with hyperthyroidism or carcinoma. This study confirms the rarity in adult Nigerians of the following tumours each of which occurred only once in the series: mediastinal, solid ovarian teratoma, testicular and sacrococcygeal teratoma. The latter two were malignant and the sacrococcygeal tumour has been reclassified as an endodermal sinus tumour. PMID- 6283862 TI - An evaluation of the New Roche Diagnostics Kit for the rapid identification of clinically important non-dextrose, non-fastidious gram-negative rods. AB - The evaluation of the new Roche Diagnostics Commercial kit (Roche Diagnostics, 1975) for the identification of non-dextrose fermenting, non-fastidious gram negative rods has been compared with conventional methods in the recognition and identification of these non-dextrose, non-fastidious gram-negative bacteria. This new kit has definite advantages over and above the conventional methods in a number of ways. It is cheaper to run, and above all it is less cumbersome, less time consuming and it is accurate. Apart from all these, the new method makes use of the odour test (Roche Diagnostics, 1975) and other additional conventional tests recommended by the Oxi/ferm manufacturer. PMID- 6283863 TI - Changes in lysosomal hydrolases from cord blood of vigorous and asphyxiated Nigerian newborn infants. AB - The activities of lysosomal hydrolases, beta-glucuronidase and acid phosphatase, were assayed in cord sera from forty-seven non asphyxiated babies and eleven babies who were severely asphyxiated at birth. Significant changes of these enzymes were observed in cord serum. Compared to the control group, serum beta glucuronidase (determined at pH 4.5 using fluorogenic substrate, 4-methyl umbelliferyl-B-D glucuronide) was 2.2 fold higher (P less than 0.01) in the asphyxiated babies. Similarly, serum acid phosphatase activity estimated by using 4-methyl-umbelliferyl-phosphate was 1.8 fold higher (P less than 0.01) in asphyxia neonatorum. The clinical relevance of this finding lies in its ultimate application, using the lysosomal hydrolase activity in scalp blood sample to evaluate objectively the foetal well-being. PMID- 6283864 TI - Chromatographic study of hexamethylene diisocyanate cross-linked polypeptides in normal Nigerians. PMID- 6283865 TI - Serum thyroglobulin measurement in various thyroidal disorders. AB - A specific double antibody radioimmunoassay for human serum thyroglobulin was established. Serum thyroglobulin concentrations were measured in euthyroid non goitrous subjects and in patients with various specific pathologies of the thyroid gland. Thyroglobulin was detectable in the serum of most euthyroid non goitrous subjects studied. The serum thyroglobulin concentrations obtained in patients with hyper or hypofunctioning thyroid glands seem to mirror gland activity in terms of hormone production, being high in thyrotoxicosis and low in hypothyroidism. Elevation of serum thyroglobulin concentrations were observed in patients with non-toxic goitres. There was a great deal of overlap in the values obtained in patients with benign goitres and those with malignant lesions making serum thyroglobulin concentration an unreliable discriminant. However, it would seem that once histological diagnosis is established serum thyroglobulin measurements could be useful in the management and follow-up of patients with differentiated primary thyroid malignancies. The likely mechanism(s) of release of thyroglobulin in different thyroidal states and the clinical potential of this new entry into thyroidology are extensively discussed. PMID- 6283866 TI - The protective potentials of 4-chloro-3, 5-xylenol (Dettol) against Mansonian schistosomiasis. AB - Laboratory tests were conducted to determine the prophylactic effectiveness of 4 chloro-3, 5-xylenol (Dettol) against mammalian schistosomiasis. The findings showed that this antiseptic was capable of protecting the albino mice against Mansonian schistosomiasis and of acting as an effective miracidicide, cercaricide and molluscicide. PMID- 6283867 TI - Hepatoblastoma: technetium sulfur colloid uptake simulating focal nodular hyperplasia. PMID- 6283868 TI - Fibrolamellar hepatocarcinoma: radiology, management, and pathology. PMID- 6283869 TI - Abdominal masses in children: multiorgan imaging with 99mTc methylene diphosphonate. AB - Technetium-99m methylene diphosphonate (99mTc MDP) imaging was performed in 29 consecutive children with abdominal masses. Dynamic images of the inferior vena cava were obtained by injecting the radiotracer in the feet. Serial renal images were obtained for the next 30 min. Routine bone imaging was performed at about 3 hr. The radionuclide studies of the inferior vena cava accurately diagnosed total obstruction and displacement to the left but not partial obstruction and displacement to the right. The abnormalities on early renal imaging included displacement (14), distortion (seven), obstruction (eight), and nonvisualization (one). All patients with Wilms tumor (eight) had either nonvisualized or distorted renal parenchyma. Patients with neuroblastoma (17) and other tumors (four) had displacement and obstruction. Soft-tissue accumulation of 99mTc MDP was noted in two Wilms tumors and 12 neuroblastomas. PMID- 6283870 TI - Computed tomographic evaluation of primary osseous malignant neoplasms. AB - A total of 128 patients with pathologically confirmed primary osseous malignant lesions was examined by computed tomography (CT). In each case, the CT findings were compared with those from the standard radiographs, tomograms, and isotope bone scans as well as with the clinical findings, in regard to tumor detection, diagnosis, and extent. Even though CT demonstrated all lesions, 96% were seen on radiographs, with only 4% of tumors identified solely by CT. In 7% of cases, CT provided unique diagnostic information not obtainable by other means. In 77% of cases, CT gave a better indication of tumor location, extent, and relationships than did any of the other methods. After treatment, CT was efficacious in the detection or ruling out of recurrences and in patient follow-up after chemotherapy or radiation therapy. PMID- 6283871 TI - Radionuclide bone scanning of osteosarcoma: falsely extended uptake patterns. AB - The pathologic specimens of 18 osteosarcomas of long bones were examined to correlate histologic abnormalities with abnormalities seen on preoperative 99mTc pyrophosphate or methylene diphosphonate bone scans. Seven scans accurately represented the extent of the tumor. Eleven scans disclosed increased activity extending beyond the radiographic abnormalities. In eight of these, there was no occult tumor extension and in the other three, the scan activity did not accurately portray the skip metastases that were present. Therefore, these 11 scans demonstrated the falsely extended pattern of uptake beyond the true limits of the tumors. Pathologic slides were available for 10 of the 11 areas of bone that exhibited extended uptake. In two instances, there was no pathologic abnormality. In the other eight cases we found marrow hyperemia, medullary reactive bone, or periosteal new bone. This is the first description of these histologic abnormalities of medullary bone in areas of extended uptake on radionuclide bone scans. PMID- 6283872 TI - Localization of the source of hyperinsulinism: percutaneous transhepatic portal and pancreatic vein catheterization with hormone assay. AB - Insulin concentrations were measured in the portal venous system through the percutaneous transhepatic approach in 12 patients with organic hyperinsulinism. Single insulinomas were found in 10, an adenoma and islet cell hyperplasia in one, and nesidioblastosis in one patient. Angiography localized adenomas in two. Local step-ups of insulin levels in the portal venous system, found in all patients with insulinomas, corresponded to tumor sites at surgery. Multiple step ups were found in the portal venous system of the patient with nesidioblastosis and the patient with an adenoma with islet cell hyperplasia. Three tumors were localized in the head of the pancreas which would indicate the primary surgical approach rather than blind resection of the body and tail. Three of the 10 adenomas were not identifiable by palpation after pancreatic mobilization during surgery. In all patients, organic hyperinsulinism was cured after surgery. Percutaneous transhepatic portal and pancreatic vein catheterization with measurement of radioimmunoactive insulin concentrations is a safe and reliable method and may play an important role in the localization of adenomas in patients with normal angiographic studies and previous negative surgical explorations. PMID- 6283874 TI - Indigestible residue or dietary fiber? PMID- 6283873 TI - Lack of effect of dietary fiber on serum lipids, glucose, and insulin in healthy young men fed high starch diets. AB - Eight healthy young men were fed a 72% carbohydrate high starch diet either high or low in dietary fiber for 4 days in a double cross-over design. Both groups showed a slight transient increase in plasma triglyceride level and a decrease in total and high-density lipoprotein cholesterol. There were few differences in glucose and insulin levels after glucose and meal tolerance tests after each diet. Fasting triglycerides and high-density lipoprotein cholesterol were inversely related at base-line; insulin response to oral glucose was inversely related to high-density lipoprotein cholesterol levels at the end of the study. We conclude that a high carbohydrate high starch diet, whether high or low in fiber, caused little increase in triglycerides, with little difference between the high and low fiber diets. Dietary fiber did not influence the fall in plasma cholesterol or high-density lipoprotein cholesterol concentrations over and above that seen after the low fiber diet. PMID- 6283876 TI - Plasma concentration of misonidazole and peripheral neuropathy. PMID- 6283875 TI - Pions, protectors. Examples of a vigorous decade in radiotherapy. PMID- 6283877 TI - Retroperitoneal synovial sarcoma. A report of four cases. AB - Synovial sarcoma most commonly affects the extremities, especially the lower thigh and knee region; other primary sites, in contrast, have been only infrequently reported. Therefore, we undertook this study of four synovial sarcomas arising in the retroperitoneum. This group, whose ages ranged from 17-57 years (mean, 36 years), comprised three males and one female. The nonspecific clinical presentations consisted of pain and/or a mass; diverse preoperative radiographic procedures merely served to reinforce the impression of a retroperitoneal mass. Histologically, all four examples revealed a typical biphasic pattern. Two cases showed a predominant spindle cell component; two others disclosed a more even distribution of the spindle-cell and epithelioid cell elements, one of these containing, in addition, broad areas of stromal calcification. Follow-up data, obtained for two patients, indicated that both died as a direct result of peritoneal sarcomatosis. The differential diagnosis of selected spindle-cell and biphasic neoplasms known to arise in the retroperitoneum will be discussed. PMID- 6283878 TI - Kawasaki syndrome in two cousins with parainfluenza virus infection. PMID- 6283879 TI - Inhibition of basal and stimulated gastric acid secretion by an enkephalin analogue. AB - The effect of a synthetic enkephalin analogue (Sandoz FF33-824: DAMME) on basal and submaximally stimulated gastric acid secretion was investigated in six healthy hypersecretors. Basal acid secretion was markedly inhibited by the enkephalin analogue. When gastric secretion was stimulated by a continuous intravenous infusion of 100 ng/kg/h of pentagastrin the enkephalin analogue reduced acid output by 50%. Intravenous naloxone reversed the subjective effects of the enkephalin analogue but did not reverse the inhibitory effect on basal and stimulated acid secretion. PMID- 6283880 TI - Sclerosing carcinoma of the right hepatic duct at the porta hepatis: anicteric presentation of early hilar cholangiocarcinoma. AB - Asymptomatic presentation of hilar cholangiocarcinoma is exceptional. We report an asymptomatic patient presenting with an abnormal alkaline phosphatase who proved to have a unilateral hilar cholangiocarcinoma obstructing the right heptic duct at the porta hepatis. Hyperbilirubinemia was absent. Subsequent to partial tumor resection and palliative decompression with right intrahepatic cholangiojejunostomy, the patient developed cholangitis with abscess formation. The efficacy of biopsy, tumor resection, and surgical decompression in hilar cholangiocarcinoma is discussed. Postmortem histology revealed biliary cirrhosis and acute cholangitis in the obstructed right liver lobe and minimal portal fibrosis in the unobstructed left liver lobe. The absence of bile stasis suggests compensatory bile excretion mechanism in the unobstructed left lobe. We believe the clinical and pathological findings in the patient to be part of the early natural history of hilar cholangiocarcinoma. Complete workup of asymptomatic alkaline phosphatase elevations in patients without previous history of hepatobiliary disease may prove fruitful in revealing incipient localized, unilateral bifurcation tumors. PMID- 6283881 TI - Viral Hepatitis: a review. PMID- 6283882 TI - Oral contraceptives and cancer of the liver: a review with two additional cases. PMID- 6283884 TI - New insights and questions about glucocorticoid-suppressible hyperaldosteronism. PMID- 6283883 TI - The effect of naloxone, morphine, and an enkephalin analogue on cholecystokinin octapeptide-stimulated gallbladder emptying. PMID- 6283885 TI - Epstein-Barr virus-induced diseases in boys with the X-linked lymphoproliferative syndrome (XLP): update on studies of the registry. AB - Analyses of 100 subjects with the X-linked lymphoproliferative syndrome (XLP) in 25 kindreds revealed four major interrelated phenotypes: infectious mononucleosis, malignant B-cell lymphoma, aplastic anemia, and hypogammaglobulinemia. Eighty-one of the patients died. Two male subjects were asymptomatic but showed immunodeficiency to Epstein-Barr virus (EBV). Seventy five subjects had the infectious mononucleosis phenotype and concurrently, 17 subjects of this group had aplastic anemia. All subjects with aplastic anemia died within a week. Aplastic anemia did not accompany hypogammaglobulinemia or malignant lymphoma phenotypes. Hypogammaglobulinemia had been detected before infectious mononucleosis in three subjects, after infectious mononucleosis in five subjects, and was not associated with infectious mononucleosis in 11 boys with hypogammaglobulinemia. In nine subjects infectious mononucleosis appeared to have evolved into malignant lymphoma; however, the majority of patients with malignant lymphoma showed no obvious antecedent infectious mononucleosis. One subject had infectious mononucleosis following recurrent malignant lymphoma. Twenty-six of 35 lymphomas were in the terminal ileum. Results of immunologic and virologic studies of 15 survivors revealed combined variable immunodeficiency and deficient antibody responses to EBV-specific antigens. Mothers of boys with XLP exhibited abnormally elevated titers of antibodies of EBV. Subjects of both sexes with phenotypes of XLP should be investigated for immunodeficiency to EBV. Persons with inherited or acquired immunodeficiency may be vulnerable to life threatening EBV-induced diseases. PMID- 6283886 TI - Herpes simplex virus: a cause for concern. AB - Herpes simplex virus (HSV) is one of several sexually transmitted agents receiving increased attention. HSV infection is a serious problem due to: the increasing number of cases; its ability to remain latent in the host and periodically cause recurrent infection; and lack of effective treatment. HSV may be acquired by infants delivered vaginally to a woman experiencing active infection. Neonatal herpes is a devastating disease, with 60% overall mortality and serious sequelae in many of the survivors. Infection by HSV is detected most reliably by virus culture, but cytology or direct antigen detection may also be useful especially when culture is not available. PMID- 6283887 TI - Isolated ACTH deficiency presenting as severe hypercalcemia. PMID- 6283888 TI - Carcinoma of the cervix--a review. AB - Carcinoma of the cervix is the fourth most common neoplasm in women. The mortality from this tumor has dropped with the advent of Papanicolaou smears and routine periodic screening, particularly in high risk populations. Diagnosis and staging includes a careful physical examination, the use of colposcopy, directed biopsy, intravenous urogram and cystoscopy. Computed tomography and lymph angiography may be helpful for detection of iliac or paraaortic lymph nodes. Early, noninvasive stages of this disease (CIN) may be treated with cryosurgery or laser vaporization. Carcinoma in situ (CIS) and microinvasive carcinoma is usually treated with simple hysterectomy for cure. More advanced invasion localized to the cervix may be treated with radical hysterectomy or radiation therapy with 90% of patients surviving 5 years. More advanced tumors are treated with external and intracavitary radiation therapy. For patients with paraaortic lymph node involvement or recurrent tumor, 5-year survival is less than 10%. Chemotherapy may provide some palliation to patients with recurrent tumors but does not increase long term survivorship. PMID- 6283889 TI - Syndactyly type V. AB - We report a mother and three of her four children with type V syndactyly. All the patients had metacarpal 4-5 fusion. The other hand anomalies consisted of abnormal origin of the fifth fingers, anomalies of digits 4 and 5, brachydactyly, syndactyly, camptodactyly, absent distal interphalangeal creases, and unusual palmar dermatoglyphics. Anomalies of the feet consisted of varus deviation of the metatarsals, valgus deviation of the toes, hyperplasia of the first ray, and hypoplasia of the third to fifth rays. None of the patients had metatarsal fusions. The anomalies were similar in the mother and her two older sons far less severe in her daughter. This daughter also had a congenital anomaly of the urinary tract. Anomalous and/or defective muscle and tendon insertions were demonstrated in one patient during an operation. Syndactyly V is transmitted as an autosomal dominant trait. PMID- 6283890 TI - Effect of conjugated estrogens on vaginal blood flow in surgically menopausal women. AB - The effect of an intravenous bolus of 25 mg of conjugated estrogen (Premarin) on vaginal blood flow in seven surgically menopausal women was measured by a thermal conductance probe. Electrical power (0.8 W) was supplied to a resistance wire incorporated in the surface of the probe, and its surface temperature was monitored continuously for 30 minutes six times daily for 3 days. The mean temperature difference (delta T) between the heated probe surface and the vagina remained unchanged during a preinjection control day. While there were individual differences in response, analysis of mean delta T during the first day following the injection of the estrogen revealed a significant linear downward trend at delta T, indicating a rise in vaginal blood flow. Delta T plateaued on day 2 and showed a slight rise in the morning of day 3. It may be concluded that estrogen administered in this dose and form markedly increases vaginal blood flow. PMID- 6283891 TI - Effect of dietary fiber on the oral glucose tolerance test in pregnancy. AB - Paired oral glucose tolerance tests (GTTs) were performed during the third trimester of pregnancy to determine whether soluble fiber, which is known to blunt the rise in postprandial serum glucose in nonpregnant subjects, demonstrated similar effects in pregnancy. Normal pregnant women (n = 14) and gestational diabetic patients (n = 4) were first given a standard 100 gm oral GTT, and serum levels of glucose and insulin were determined in the fasting state and at 60, 120, and 180 minutes after ingestion of the glucose load. Several days later, a second GTT was done. In this test, glucose was combined with 15 gm guar gum, a purified soluble dietary fiber. Improved glucose tolerance was documented in all but one patient. Guar gum produced a significant reduction in mean serum glucose levels at 1 hour (p less than 0.005), 2 hours (p less than 0.005), and 3 hours (p less than 0.005), and each of the four gestational diabetic patients tested showed a normal glucose tolerance curve. The soluble dietary fiber, guar gum, did limit the postchallenge rise in serum glucose in pregnant subjects. Furthermore, the improvement in glucose tolerance in gestational diabetic patients is sufficient to suggest a potential clinical value for dietary fiber in the treatment of gestational diabetes. PMID- 6283893 TI - A new understanding of ocular herpetic disease. PMID- 6283892 TI - Expression of herpes simplex virus type 2 antigens in premalignant and malignant human vulvar cells. AB - Vulvar biopsies and explants from patients with vulvitis, hyperkeratosis, condyloma acuminatum, severe dysplasia, and squamous cell carcinoma were screened for herpes simplex virus type 2 (HSV-2) antigens. Immunoperoxidase staining for VP143, an early nonstructural polypeptide of HSV-2, was identified within three biopsies exhibiting severe dysplasia, three of seven with carcinoma in situ, and one of three with invasive squamous cell carcinoma. Similar staining for VP143 was observed in derived explants. Staining for VP119, the major envelope glycoproteins of HSV-2, was identified within tissues which were positive for VP143. Both proteins were expressed in the absence of staining for virus capsid proteins, detection of virus structures by electron microscopy, and isolation of infectious virus by co-cultivation, indicating that only a fragment of the virus genome was expressed. Neither VP143 nor VP119 was identified in biopsies exhibiting vulvitis, hyperkeratosis, or condyloma acuminatum. These data indicate a close relationship between HSV-2 and vulvar neoplasia. PMID- 6283894 TI - Carpal tunnel syndrome. Measurement of sensory potentials using ring and index fingers. PMID- 6283895 TI - Glucocorticoids and hypothyroidism modulate development of fetal lung insulin receptors. AB - We investigated the development of insulin receptors in membranes of fetal rabbit lung during normal ontogeny and the effect of glucocorticoids and hypothyroidism. Specific binding of 125I-insulin to fetal lung membranes increased progressively to a peak at 29 days gestation, declining by 30 days. Scatchard plots were curvilinear and revealed a progressive increase in receptor numbers (X 10(10)/mg protein) from 129 +/- 7 (mean +/- SE) at 22-24 days to 575 +/- 16 at 29 days, declining to 467 +/- 12 at 30 days, term being approximately 31 days. Affinities did not change throughout gestation and were similar to those of adult lung; receptor numbers in adults were significantly lower than in fetuses at 26-30 days. Epinephrine and PGE1 could evoke a doubling of cAMP production in adult and fetal lung membranes until 29 days. Concomitantly with the fall in fetal insulin receptor number at 30 days, cAMP production in response to epinephrine or PGE1 increased fivefold. Induction of fetal hypothyroidism decreased insulin receptor numbers in the lung of the 28-day fetus by 70% from control (P less than 0.001) without a change in receptor affinity. In contrast, betamethasone administration increased fetal lung insulin receptor numbers by 250% (P less than 0.001) but did not alter their affinity; maternal lung insulin receptors were not altered. Thus, normal ontogeny of the fetal lung insulin receptor is characterized by a progressive increase in number followed by decline immediately before parturition associated with a sharp increase of cAMP responsiveness of the membranes. Hypothyroidism and glucocorticoid exposure can modulate the normal development of the fetal lung insulin receptor. PMID- 6283896 TI - Estradiol stimulation of LH response to LHRH and LHRH binding in pituitary cultures. AB - The relationship between 17 beta-estradiol (E2) stimulation of luteinizing hormone (LH) response to LH-releasing hormone (LHRH) and E2 effect on LHRH binding was examined in pituitary monolayer cultures prepared from female rats. E2 pretreatment significantly (P less than 0.05) augmented the LHRH-induced LH release to 158-180% of the non-E2-treated controls. The maximal E2-priming effect could be observed after 1 day of treatment. E2 treatment for 3 days stimulated [D Ala6]luteinizing hormone-releasing hormone (LHRHa) binding to about 1.5-fold that of the non-E2-treated controls without affecting the dissociation constant of LHRH receptor (Kd = 4 X 10(-10) M). The stimulatory effect of E2 on cell proliferation as determined by [3H]thymidine incorporation was also observed 3 days after treatment. However, E2 stimulation of LH accumulation in the cultured cells could be detected as early as 4 h after treatment. These results indicate that E2-priming effect on pituitary LH response to LHRH is initially associated with an increase in cellular LH content and later associated with increases in LHRH binding and in an index of cell proliferation that may include the LH producing cells. PMID- 6283897 TI - Calcium homeostasis in chronic streptozotocin-induced diabetes mellitus in the rat. AB - Calcium homeostasis was studied in freely fed control, streptozotocin diabetic, long-term and short-term insulin-treated diabetic rats 7 wk after the induction of diabetes. In contrast to the short-term (5-12 day) diabetic rat model, intestinal absorption of calcium was markedly enhanced in chronically insulin deficient animals. Moreover, conventional balance studies showed that these animals were in positive calcium balance despite severe hypercalciuria. Intestinal hyperabsorption of calcium in long-standing diabetic rats occurred despite low levels of circulating 1,25-dihydroxyvitamin D and hypercorticosteronism and was attended by hypercalcemia and suppression of both plasma parathyroid hormone (PTH) and urinary cyclic 3',5'-AMP (cAMP). Long-term insulin replacement completely normalized the intestinal hyperabsorption of calcium, corrected the plasma calcium, and significantly increased circulating PTH and urinary cAMP excretion. Insulin therapy also corrected the decreased plasma 1,25-dihydroxyvitamin D observed in untreated diabetic animals. Intestinal hyperabsorption of calcium appeared to be only partially corrected by short-term insulin therapy. The accumulated results reveal decided differences in calcium homeostasis and hormonal response between the rats with long-standing diabetes and those with diabetes of short duration. PMID- 6283898 TI - NaCl entry mechanisms in the luminal membrane of the renal tubule. PMID- 6283899 TI - Systemic effects of NaHCO3 in experimental lactic acidosis in dogs. AB - Lactic acidosis is characterized by metabolic acidosis due to accumulation of H+ ions from lactic acid with blood lactate of at least 5 mM. The standard treatment is intravenous NaHCO3, with resultant mortality in excess of 50%. Despite the high mortality, the metabolic and systemic effects of NaHCO3 used in the treatment of lactic acidosis have not been extensively studied. The present experiments in diabetic dogs were designed to address these questions. Dogs with phenformin-induced lactic acidosis (blood lactate above 5 mM, arterial pH below 7.20) were treated with equimolar amounts of either NaCl or NaHCO3 or received no therapy. Intravenous NaHCO3 resulted in a decline of cardiac output and intracellular pH (pHi) of liver and erythrocytes, whereas treatment with NaCl did not. With NaHCO3 but not with NaCl infusion gut lactate production increased almost stoichiometrically, with no change in arterial pH or bicarbonate but with a doubling of lactate. Bicarbonate also resulted in a decrease of hepatic portal vein blood flow. The mean survival time and percent mortality were similar in NaCl- vs. NAHCO3(-) treated animals. Although both groups lived longer than did animals receiving no therapy, the differences were not significant. Thus, treatment of experimental lactic acidosis with either NaCl or NaHCO3 or with no therapy results in no change of blood pH and bicarbonate and in a similar mortality. In terms of systemic effects, however, NaHCO3 results in significant decrements of liver and erythrocyte pHi, hepatic portal vein blood flow, and cardiac output and in significant increments of gut lactate production, whereas NaCl does not. The data suggest that the rationale for therapy of lactic acidosis with NaHCO3 should probably be reevaluated. PMID- 6283900 TI - Specialized function of carbonic anhydrase-rich and granular cells of turtle bladder. AB - The mucosal surface of the turtle bladder consists of two major cell types, carbonic anhydrase-rich (CA) and granular (G) cells. These cells types may have specific transport functions. To separate and study these cells, they were removed from the bladder by collagenase digestion and then separated into two distant subpopulations by Ficoll discontinuous density-gradient centrifugation. By morphological criteria the lighter subpopulation (band II) consists primarily of CA cells and the denser population (band III) G cells. The distribution of two enzymes important for H+ transport, CA and glucose-6-phosphate dehydrogenase (G-6 PD), and the effect of specific transport inhibitors, acetazolamide and ouabain, on O2 consumption of these cells was examined. The CA activity of band II cells was 4.5 times greater than that of band III cells. The G-6-PD activity of band II cells was 2.2 times greater than that of band III cells. The rate of O2 consumption of band II cells, 4.73 microliters.mg protein-1.h-1, was reduced 40% by 5 X 10(-5) M acetazolamide but not by ouabain. Band III O2 consumption, 3.43 microliters.mg protein-1.h-1, was reduced 30% by 10(-4) M ouabain but not by acetazolamide. On the basis of the distribution of CA and G-6-PD and the effect of acetazolamide and ouabain on O2 consumption, we propose that CA cells are primarily responsible for H+ transport and G cells for Na+ transport. PMID- 6283901 TI - Permeability and channel structure of reptilian skin. AB - A study of the permeability of shed epidermis from some terrestrial and freshwater snakes was conducted. Permeability to Br, Na, and K ions was very low and showed a higher influx than efflux in most cases. Permeability to the smaller water molecule was much greater, and in contrast efflux was higher than influx. Skins from aquatic snakes with larger water permeabilities also showed greater permeabilities to Na and K. The highly aquatic Regina septemvittata has the most permeable skin of any snake. Ethanol fluxes were higher than expected for a tracer of its size, perhaps due to its solubility in lipid. Na fluxes through whole live skins of Nerodia cyclopion floridana in vitro were not significantly different from those of the shed skin alone. Isolated hinge regions showed complete water impermeability, suggesting that channels through the skin are located only in the scale region. Dermal water efflux into dry air was considerably less than water-to-water efflux. Lipid extraction increased permeability markedly and eliminated the asymmetry of water and ion fluxes. Lipid replacement with linoleic acid restored half of the water impermeability lost during extraction. Protein extraction did not significantly increase membrane permeability but did eliminate the permeability difference between dry and hydrated skins. Two sizes of lipid-lined channels extending through a protein matrix are suggested as a possible model for snake skin. The diameter of the channels apparently varies in relation to the differing water, Na, and Cl concentrations on opposite sides of the skin, but the mechanism of this adjustment is unknown. PMID- 6283902 TI - Selective inhibition by epinephrine of parathyroid hormone-stimulated adenylate cyclase in rat renal cortex. AB - Parathyroid hormone- (PTH) stimulated adenylate cyclase activity in homogenates of rat renal cortex was inhibited by l-epinephrine. The specificity of the inhibition indicated that it was mediated by alpha 2-receptors. The inhibition of PTH-stimulated activity was greater than the inhibition of basal activity. The absolute decrease in adenylate cyclase activity produced by 10-4 M l-epinephrine was from 16.3 +/-0.6 (SE) to 11.2 +/- 0.6 pmol.min-1.mg-1 for activity stimulated by 10 microgram/ml PTH. Basal activity was decreased from 2.3 +/- 0.07 to 1.7 +/- 0.04. A similar inhibition of PTH-stimulated adenylate cyclase by l-epinephrine was demonstrated in preparations of renal cortical tubules. In contrast, the quantitative decrease in vasopressin-or calcitonin-stimulated activity by 10-4 M l-epinephrine was the same as the decrease in basal activity. These results demonstrate that PTH receptors that stimulated adenylate cyclase and alpha 2 adrenergic receptors that inhibit adenylate cyclase are present on the same cells in the renal tubules. Thus, a mechanism exists whereby alpha-adrenergic agonists can oppose the tubular actions of PTH via a direct inhibition of adenylate cyclase activity. PMID- 6283903 TI - Na+/H+ antiporter of brush border vesicles: studies with acridine orange uptake. AB - Fluorescence quenching of acridine orange was used to characterize the generation and collapse of pH gradients by the Na+/H+ antiporter of brush border membrane vesicles prepared from rabbit renal cortex. Quenching was observed when acridine orange, a weak base, was taken up to an acidic intravesicular space. Na+/H+ exchange was examined with both Na+ uptake and efflux studies. Acridine orange fluorescence quenching demonstrated the cation specificity of the Na+/H+ antiporter (i.e., sodium and lithium) and was inhibited by amiloride. Parallel studies with nigericin, a K+/H+ antiporter, demonstrated that acridine orange responded very rapidly to pH gradients. Therefore, acridine orange equilibration was not rate limiting in our studies of the Na+/H+ antiporter. Initial rate measurements were made to obtain kinetic parameters for the Na+/H+ antiporter. In sodium influx studies, the half-maximal rate of acridine orange fluorescence change was obtained with an external sodium concentration of 13.3 +/- 0.5 mM. PMID- 6283904 TI - Kallikrein-kinin system in regulation of submandibular gland blood flow. PMID- 6283905 TI - Myocardial aging: functional alterations and related cellular mechanisms. AB - Studies that have examined the effect of age on the myocardium do not support the notion that a generalized decline in myocardial function occurs with adult aging but, rather, that certain specific changes in cardiac biochemistry and function occur. Although in many instances the molecular bases for these changes have not been precisely defined, sufficient clues regarding their nature are at hand to provide at least the first step toward an understanding of the dramatic effects of aging or time on the myocardium. A fundamental understanding of the effects of aging will provide an additional dimension to studies of myocardial biochemistry and function and may be useful in elucidating mechanisms of excitation contraction coupling in the heart. PMID- 6283906 TI - 2-Deoxy-D-glucose uptake by rat granular pneumocytes in primary culture. AB - Uptake of 2-deoxy-D-glucose (DG) was investigated with rat granular pneumocytes isolated in primary culture. Cells attached to flasks were incubated in Minimal Essential Medium usually containing 5 mM DG in place of glucose. Uptake of DG increased progressively with time of incubation and approached a plateau value of 35-40 mumol/10(6) cells at 60 min. Uptake increased as a function of external DG concentration with half-maximal uptake at approximately 2.0 mM DG. DG uptake was inhibited by the presence of glucose, alpha-methylglucoside, phlorizin, ouabain, or sodium-free medium. After 60 min incubation, approximately 20% of total intracellular DG was in the free form, and the calculated mean intracellular concentration of free DG (n = 4) was approximately twice the external concentration. Phosphatase activity was indicated by increase in free DG and efflux from cells after removal of external DG. In comparison with pneumocytes, uptake of DG by alveolar macrophages showed different kinetics, and intracellular free DG did not exceed the extracellular concentration. These findings indicate that type II cells take up DG by a sodium-dependent, carrier-mediated transport process that results in accumulation of free sugar against a concentration gradient. PMID- 6283907 TI - Surfactant secretion: evidence that cholinergic stimulation of secretion is indirect. AB - There is strong evidence that cholinergic agents stimulate the secretion of surfactant in vivo and in the isolated perfused lung and that they do not stimulate surfactant secretion in isolated type 2 alveolar cells. These observations suggest that in multicellular systems the cholinergic effect is indirect. In the present work we have accrued the following support for this hypothesis. 1) Propranolol blocked the in vivo stimulation of disaturated phosphatidylcholine (DSPC) secretion by pilocarpine. 2) Bilateral adrenalectomy decreased by 50% the in vivo stimulation of DSPC secretion by pilocarpine. 3) Bilateral vagotomy did not block the increased secretion of DSPC produced in vivo by periodic deep inflations. 4) Pilocarpine (10(-7) M) stimulated DSPC secretion in the isolated perfused lung, and this effect was blocked by indomethacin as well as by atropine. We conclude that cholinergic stimulation of the secretion of surfactant in rats is indirect, i.e., cholinergic agonists do not stimulate the secretion of surfactant by acting directly on type 2 alveolar cells. PMID- 6283908 TI - Modulation of vasopressin action by reducing agents in Bufo marinus. AB - The effects of the reducing agent dithiothreitol (DTT) on vasopressin (AVP) stimulated osmotic water flow and adenylate cyclase activity were studied in the urinary bladder of Bufo marinus. DTT produced concentration-dependent inhibition of the hydroosmotic water permeability response to 10 mU/ml AVP and 10 mM theophylline but did not inhibit the response to 10 mM adenosine 3',5'-cyclic monophosphate (cAMP). The inhibitory effects of DTT on AVP responsiveness were partially reversed by washing in DTT-free Ringer solution or by addition of oxidizing agents such as dehydroascorbic acid (DHA) or H2O2. The inhibitory effects of DTT were completely reversed by washing in DTT-free Ringer plus addition of DHA. In addition, the inhibitory effects of DTT on AVP-induced osmotic water flow were partially reversed by the GTP analogue 5'-guanylyl imidodiphosphate [Gpp(NH)p]. DTT also inhibited the adenylate cyclase response to AVP but did not alter the response to AVP plus Gpp(NH)p or the response to NaF. These observations suggest that the inhibitory effect of thiol compounds on AVP responsiveness may be modulated through alterations of a redox system distal to the hormone receptor but proximal to the catalytic subunit of adenylate cyclase. Inasmuch as Gpp(NH)p partially reversed the inhibitory effects of DTT on AVP stimulated osmotic water permeability and prevented the inhibitory effect of DTT on AVP-stimulated adenylate cyclase, an effect on either GTPase or binding of GTP to the regulatory protein of adenylate cyclase is suggested by these observations. PMID- 6283909 TI - Effects of alterations in calcium levels on cat small intestinal slow waves. AB - Intact segments of cat intestinal muscle and strips of isolated longitudinal muscle were treated with agents that reduce intracellular calcium concentration: incubation in 0-calcium saline, treatment with calcium conductance blockers, elevated extracellular magnesium concentration, or alkalinization with NH4Cl. These treatments reduced amplitude and frequency of slow waves in intact segments but only reduced frequency in isolated longitudinal muscle. The reduction in frequency was characterized by prolongation of the hyperpolarized phase of the slow waves. Treatments that would moderately increase intracellular calcium concentration, i.e., increasing external calcium to four times normal levels or lowering pH by CO2, increased slow-wave frequency. Increased frequency was associated with reduced amplitude and shortening of the hyperpolarized phase of the slow waves. Greater than four times normal calcium levels and intense spiking reduced slow-wave frequency. Chlorotetracycline fluorescence, an indicator of intracellular calcium concentration, showed fluctuations synchronous with slow waves. It is concluded that the reactions that pace the generation of slow waves are dependent on the level of intracellular calcium. PMID- 6283910 TI - Effects of dantrolene and D2O on K+-stimulated respiration of skeletal muscle. AB - We have examined the effects of dantrolene and D2O on the K+-stimulated respiration in frog skeletal muscle. The threshold for K+ stimulation was around 10 mM extracellular potassium concentration ([K+]o). A further marked increase in respiration to levels about ten times the resting level was noted when [K+]o was between 15 and 20 mM. The increase was sustained for hours when [K+]o was less than 20 mM; however, with higher concentrations the stimulation consisted of an initial burst followed by a decline. Dantrolene shifted the relationship between [K+]o and peak increase in respiration toward higher [K+]o by about 10 mM; in addition it nearly completely blocked the sustained component of the increase. D2O, nearly abolished the K+-induced respiration. Neither agent shifted the relationship between [K+]o and membrane potential nor abolished the stimulation of respiration caused by caffeine. Dantrolene did not block the stimulation of Na+ efflux caused by 15 mM K+. The results with these agents are consistent with the proposal that K+-stimulated respiration is due to Ca2+ release into the cytoplasm. In addition, they provide evidence that the stimulated rate of Ca2+ release into the cytoplasm can remain at a persistently high level for hours provided [K+]o does not exceed 20 mM. We calculated that the level of this constant Ca2+ release is about 3.4 X 10(16) ions/(s.cm3). PMID- 6283911 TI - Deficiency of hormone receptor-adenylate cyclase coupling protein: basis for hormone resistance in pseudohypoparathyroidism. AB - Pseudohypoparathyroidism is an inherited disorder associated with resistance to the action of several hormones, including parathyroid hormone, thyroid stimulating hormone, follicle-stimulating hormone, and luteinizing hormone. The disorders described under this designation are heterogeneous in regard to the underlying genetic defects, the phenotypic manifestation, and the severity of the defects in hormone action. The majority of affected individuals who also have the characteristic skeletal changes (heredity osteodystrophy) have a defect in the guanine nucleotide regulatory protein (G protein) that is essential for coupling certain cell-surface hormone receptors to the adenylate cyclase system. This defect is probably the cause for resistance to the action of multiple hormones. In the remaining patients the cause for hormone resistance has not been identified. PMID- 6283912 TI - Mutations that affect membrane receptor for LDL are useful for studying normal receptor function. AB - Low-density lipoprotein (LDL), the major plasma cholesterol transport protein, is taken up by cells through a receptor-mediated process. After internalization through specialized segments of the cell surface called coated pits, the LDL is degraded in the lysosome and the released cholesterol is used by cells to meet various metabolic needs. The discovery of the LDL receptor and the studies of its function have provided new insights into both the biochemical aspects of cholesterol metabolism and the cell biology of receptor-mediated endocytosis. Of paramount importance in all of these studies has been the availability of human cells that express one or more allelic mutations that affect the function of the LDL receptor. These mutations have been valuable for assessing normal receptor function. Just as important, these mutations have been used as a reference point in the development of various cytochemical and biochemical techniques for studying receptor activity. PMID- 6283913 TI - Calcium channel and prolactin release in rat clonal pituitary cells: effects of verapamil. AB - Effects of verapamil on membrane electrical properties and prolactin release were studied in a rat anterior pituitary cell line GH3. Thyrotropin-releasing hormone (TRH), Ba2+, and high concentration of K+ enhance the release of prolactin from GH3 cells. These stimulatory actions on prolactin release were inhibited by adding 10(-4) M verapamil to the bathing mediums. The maximum rate of rise of the Ca action potential was reduced to 17% of the control by addition of 10(-4) M verapamil. Ba2+ caused a sustained membrane depolarization because Ba2+ goes through the Ca channels and blocks the development of the delayed rectification. This effect of Ba2+ was also inhibited by verapamil. Verapamil suppressed both the Na+ and outward K+ currents in addition to the Ca2+ current. The suppressive effect of verapamil on the voltage-sensitive Ca current is probably responsible for the inhibition of TRH- and high K+-stimulated prolactin release because the suppression of the Na+ and outward K+ currents does not inhibit the stimulatory actions of these secretagogues. PMID- 6283914 TI - Proximal cell K+ activity: technical problems and dependence on plasma K+ concentration. AB - K+-selective liquid ion-exchanger double-barreled microelectrodes were used to measure intracellular K+ activity (alpha Ki) of proximal tubular cells in Necturus kidney. SEveral methodological problems inherent in the construction and in vivo application of such microelectrodes were considered, in particular the identification of leaky impalements and the correct assessment of alpha Ki during reversible perfusion of peritubular capillaries from control to a test solution. Peritubular potassium concentration (CpK) could be altered by means of double barreled micropipettes containing a physiologic and a high or low K solution. The control alpha Ki value averaged 57.9 +/- 8.5 mM (mean +/- SD, n = 38) and the basolateral membrane potential was -70.4 +/- 5.8 mV. Thus, the K+ equilibrium potential across the basolateral membrane (EK) was -81.1 +/- 3.4 mV. Increasing CpK from 3.0 to 30.0 mM raised alpha Ki by 7.0 +/- 1.5 mM, and lowering CpK to 0.3 mM decreased alpha Ki by 9.2 +/- 2.9 mM. CONCLUSIONS: 1) the steady-state K+ distribution across the basolateral membrane indicates that potassium is actively pumped into the cell. 2) The relative stability of alpha Ki in the face of large CpK variations underscores the effectiveness of cell homeostasis; passive mechanisms appear to contribute to this end. PMID- 6283915 TI - Temperature-induced conversion of the renal adrenoreceptor: modulating renin release. AB - The release of renin from dog cortical kidney slice preparations incubated in a physiological salt solution can be modulated by alpha- and beta-adrenergic drugs. When given to slices maintained at 37 degrees C, the beta-agonists isoproterenol (ISP) and norepinephrine stimulated renin release from the slices. When the slices were maintained at 20 degrees C, the beta-agonists had no effect on renin release. However, the alpha-agonist phenylephrine inhibited renin release from the slices incubated at 20 degrees C in a dose-dependent manner, whereas its effect on slices incubated at 37 degrees C was less pronounced. The change in response of the slices from beta dominant at 37 degrees C to alpha dominant at 20 degrees C appeared to be a receptor phenomenon. When the cortical slices were incubated with the irreversible alpha-antagonist phenoxybenzamine (POB) at 20 degrees C for 1 h, they were unable to respond to ISP when returned to 37 degrees C. However, POB had no effect on the response of slices to ISP when given at 37 degrees C. It appears that with a decrease in temperature the renal beta receptors demonstrate properties normally associated with alpha-receptors, namely the potential to be blocked by POB. This may be due to an interconversion of the renal alpha- and beta-adrenoceptors. PMID- 6283916 TI - cGMP modulation of ileal ion transport: in vitro effects of Escherichia coli heat stable enterotoxin. AB - Diarrheagenic strains of Escherichia coli have been shown to produce a heat stable enterotoxin (ST) that simulates guanylate cyclase, increases short-circuit current (Isc), and inhibits active Cl absorption in the intestine. In rabbit ileum, the ion transport effects are smaller than those produced by cAMP-related agonists. Because ST may be a selective cGMP agonist, we further explored its mode of action in rabbit ileum. ST inhibits net Na and net Cl absorption. ST also inhibits the same fraction of Cl influx across the brush border that theophylline inhibits. At maximal doses, ST and 8-bromo-cGMP (8-Br-cGMP) had nearly equal, nonadditive effects of Isc that were about 66% of that produced by 8-Br-cAMP. ST increased mucosal cGMP concentration 16-fold, whereas epinephrine, an inhibitor of secretion, increased cGMP concentration by only 30%. This is insufficient to alter ion transport because doses of ST that increased cGMP concentration by 100% failed to alter Cl fluxes. Furthermore, epinephrine did not increase cGMP concentration in isolated enterocytes. We conclude that 1) cGMP mediates ST effects on ion transport, and 2) although ST and cAMP-related agonists have the same antiabsorptive effects, ST is less effective in stimulating electrogenic Cl secretion. PMID- 6283917 TI - Effects of inhibitors of Na+-coupled ion transport on bile acid uptake by isolated rat hepatocytes. PMID- 6283918 TI - Coronary blood flow changes following activation of adrenergic receptors in the conscious dog. AB - The role of coronary vascular adrenergic receptors in changing coronary flow was studied in dogs instrumented to measure left circumflex artery blood flow (CBF), mean coronary artery blood pressure (CBP), and heart rate (HR). Norepinephrine (NE), isoproterenol (IP), and phenylephrine (PH) were injected into the left circumflex artery before and after selective intracoronary alpha- and beta 1- or combined beta 1- and beta 2-receptor blockade. NE, IP, and PH did not alter CBP (112 +/- 6 mmHg). In addition, IP and PH did not affect HR (103 +/- 4 beats/min). NE increased HR to 150 +/- 6 beats/min, which was eliminated by blocking beta 1 receptors with atenolol and by removing the left stellate ganglion. IP increased CBF from 65 +/- 9 to 115 +/- 16 ml/min (mediated by both beta 1- and beta 2 receptors). PH caused an alpha-receptor-mediated coronary vasoconstriction (42 +/ 5 ml/min), which was potentiated by beta 1- and beta 2-receptor blockade. NE caused a biphasic flow response. CBF initially increased to 117 +/- 14 ml/min (mediated predominantly by beta 1-receptors) followed by a prolonged decrease to 54 +/- 7 ml/min (mediated by alpha-receptors). Removing the left stellate ganglion did not affect the CBF response to NE. These data indicate that PH directly stimulates coronary alpha-receptors and IP stimulates myocardial beta 1- and coronary beta 2-receptors. NE also stimulates myocardial cells causing a reflex that increases HR and indirectly increases CBF. The vasoconstriction to NE and PH was not evident after pentobarbital anesthesia, whereas the coronary vasodilation and increase in HR to NE was still present. PMID- 6283919 TI - Action of preoptic injections of beta-endorphin on temperature regulation in rabbits. AB - Male New Zealand White rabbits, Oryctolagus cuniculus, were stereotaxically implanted with a guide tube above the preoptic/anterior hypothalamus area (PO/AH) for the injection of the opioid peptide, beta-endorphin (beta-E), naloxone, sodium salicylate, or physiological saline. PO/AH and ear temperature, oxygen consumption, and evaporative heat loss (EHL) were recorded in free-moving rabbits before and after injection of saline followed with beta-E, naloxone, or sodium salicylate at ambient temperatures (Ta) of 2-31 degrees C. A 5-micrograms injection of beta-E promoted a rapid reduction in ear temperature followed by a prolonged rise in PO/AH (body) temperature. Preinjection with an isovolumetric amount of the opiate antagonist, naloxone, inhibited the thermoregulatory effects of beta-E. The beta-E-induced rise in body temperature was directly correlated with Ta. beta-E had no effect on oxygen consumption at Ta's of 5 and 27 degrees C. When measured 30 min after injection, beta-E demonstrated a significant inhibition of EHL at Ta's of 27 and 31 but not 5 degrees C. The beta-E-induced rise in body temperature was not antagonized with preinjections of sodium salicylate in the PO/AH. These data indicate that beta-E promotes a regulated increase in body temperature. The mechanism of activation appears to be distinct from that of an infectious fever. PMID- 6283920 TI - Peptidergic regulation of stress-induced eating. AB - By use of the model of stress-induced (mild tail pinch) eating we have examined the interrelationships of peptides and monoamines responsible for regulating this behavior. We have shown that the synthetic opiate analog, D-Ala2-Met enkephalinamide (1 microgram), when administered intracerebroventricularly (icv) reverses the suppressive effects of the serotonin agonist, quipazine (40 micrograms icv), the beta-agonist, isoproterenol (40 micrograms icv), and the alpha-antagonist, phentolamine (150 micrograms icv), and partially reversed the effects of atropine (2.5 mg/kg sc) and the dopamine antagonist haloperidol (0.5 mg/kg sc). The opiate antagonist naloxone (10 mg/kg sc) suppressed tail-pinch induced eating, and this effect could not be reversed by the GABA-agonist muscimol (500 ng icv) nor norepinephrine (20 micrograms icv). The putative satiety hormones cholecystokinin-octapeptide (5 micrograms/kg sc) and bombesin (5 micrograms/kg sc) suppressed stress-induced eating. The suppressive effect of these substances was reversed by a number of known appetite stimulants viz., D Ala2-Met-enkephalinamide, diazepam, muscimol, and propanolol. Norepinephrine reversed the suppressive effect of bombesin but not that of cholecystokinin. Based on these results we present a hypothetical model to partially explain the peptidergic-monoamine regulation of stress-induced eating. PMID- 6283921 TI - Endurance training attenuates stress hormone responses to exercise in fasted rats. AB - Endurance exercise training produces major adaptations in hormonal and metabolic responses to exercise. This study was designed to determine whether the differences in hormone response persist in the fasted condition when liver glycogen is depleted. Rats were run on a motor-driven rodent treadmill 5 days/wk for periods up to 2 h/day for 10 wk. Trained and nontrained rats were then fasted 24 h and were run for periods ranging from 0- to 60 min. At the end of 60 min of exercise muscle glycogen was higher in trained rats (2.9 +/- 0.3 vs. 1.1 +/- 0.1 mg/g). Blood glucose was maintained at higher levels in trained rats throughout the course of the exercise (3.2 +/- 0.1 vs. 2.3 +/- 0.1 mM after 60 min). Plasma concentrations of glucagon and epinephrine increased in both groups during the exercise but were significantly lower in trained animals. Differences between trained and nontrained animals in stress hormone responses to exercise persist in the fasted state and appear to be a consequence of the capacity of trained animals to maintain higher blood glucose levels. PMID- 6283923 TI - Radiation therapy, local tumor control, and prognosis in bronchogenic carcinoma: current status and future prospects. AB - While the overall prognosis for cure of bronchogenic carcinoma remains poor for most patients, there is a growing body of evidence suggesting that rationally optimized local therapy may benefit a significant subset of patients. Local therapy in this context includes any systemic therapy (such as chemotherapy or immunotherapy) that enhances local tumor control in the chest. Compared with many other human epithelial cancers, the total local tumor burden is large for many nonresectable lung cancers and not within the tolerance for control by radiation alone. Thus there is growing evidence that combined surgery and radiation treatment will improve results, especially in the differentiated tumors. Proper selection of patients is important and must include histologic stratification in addition to conventional TNM staging. It is projected that much useful research can be conducted during this decade using clinical tools now available and those that are being tested in early clinical trials throughout the world. Likely candidates for such improvements are both oxic and hypoxic radiosensitizing drugs that should decrease the death rate from uncontrolled local cancer in the chest. PMID- 6283922 TI - Analysis of the actions of angiotensin on the central nervous system of conscious dogs. AB - Angiotensin II (ANG II) acts on the brain to elevate blood pressure (BP), stimulate drinking, increase the secretion of vasopressin and corticotropin (ACTH), and inhibit the secretion of renin. The present studies were designed to evaluate the possible physiological significance of these effects. The experiments were performed in conscious dogs with small catheters chronically implanted in both carotid and both vertebral arteries. ANG II was infused into both carotid or both vertebral arteries in doses of 0.1, 0.33, 1.0, and 2.5 ng.kg 1.min-1. Intravertebral ANG II produced dose-related increases in BP that were generally accompanied by increases in heart rate. Intracarotid angiotensin also increased BP but did not change heart rate. Intracarotid ANG II stimulated drinking and, at the highest dose only, increased the secretion of vasopressin, ACTH, and corticosteroids. Intravertebral and intracarotid ANG II suppressed plasma renin activity (PRA). In a parallel series of experiments, the effects of intravenous ANG II, in doses of 2, 5, 10, and 20 ng.kg-1.min-1, were studied. These infusions produced dose-related increases in BP and water intake and suppressed PRA. Only the highest dose of ANG II increased vasopressin or corticosteroid secretion. Analysis of these results in terms of calculated or measured changes in plasma ANG II concentration indicate that the central cardiovascular and dipsogenic actions of angiotensin, as well as the suppression of PRA, can be elicited by concentrations of the peptide that are within the physiological range. On the other hand high, probably supraphysiological, levels of ANG II are required to increase vasopressin or ACTH secretion. PMID- 6283924 TI - When are electron microscopy, histoculture, and other procedures useful in the clinical evaluation of lung cancer? AB - The earlier pessimistic outlook on the prognosis of lung cancer has had two particularly adverse effects on progress in pathologic interpretation. There has been little incentive to resolve several basic problems, and little encouragement from clinical investigators to apply electron microscopy, tissue culture, immunopathologic tests, and biochemical assays. However, substantial advances in the clinical detection and management of lung cancer have renewed the clinician's interest in pathologic interpretation. In fact, the pathologist is under increasing pressure to narrow the gap between the diagnosis that routine laboratory examination provides and the more detailed and precise interpretation that can be obtained from a research facility. This report presents recommendations for improving the level of interpretation of the routine diagnostic work-up through modest investments in newer methods, and thorough special tissue processing in anticipation of a need for assistance from the research laboratory. PMID- 6283925 TI - Carcinoma of the lung: what can be done if the carina is involved? AB - When carcinoma of the lung invades the carina, it is by definition a stage III lesion and frequently incurable. However, when lymph node invasion does not preclude resection for cure and when there are no other contraindications to such resection, techniques are now available for resection of the carina and primary reconstruction. While tracheal sleeve pneumonectomy is the operation most frequently employed for invasion of the carina by bronchogenic carcinoma that is otherwise operable, occasionally the lower lobe may also be saved. When the carina is involved by a primary neoplasm of the airways, primary resection with carinal reconstruction with or without various amounts of pulmonary resection is clearly indicated when possible. PMID- 6283926 TI - Management of tracheal tumors. AB - Since 1962, 110 primary tracheal tumors have been seen including 43 squamous cell carcinomas, 38 adenoid cystic, and 29 varied. Sixty-nine patients underwent resection of their primary tumors; an additional 33 patients with secondary tumors involving the trachea also underwent resection. Seventy-three of these 102 patients underwent primary reanastomosis. In 17, laryngotracheal resection was required; staged resections were done for the rest. Survival data indicate that benign tumors are cured by resection with reconstruction and that patients with squamous cell carcinoma, adenoid cystic carcinoma, and other types of malignant lesions obtained either long-term palliation or cure if surgical resection was possible. In patients with selected types of secondary tumors, resection and reconstruction provide prolonged palliation. PMID- 6283927 TI - The course and relations of the hypoglossal nerve and the occipital artery. AB - Three main types of variations in the relations of the occipital artery and the hypoglossal nerve found in 42 dissected specimens are described, according to the level at which the nerve crosses the external carotid artery and the point of origin of the occipital artery. In Type I, the hypoglossal nerve crosses the external carotid artery inferior to the origin of the occipital artery; in Type II, the nerve crosses the external carotid artery at the level of origin of the occipital artery; and in Type III, it crosses superior to that level. In Type III the occipital artery makes a loop around the hypoglossal nerve and is in a position to pull and exert pressure on the nerve. This possibility should be taken into consideration in the diagnosis of peripheral paresis or paralysis of the tongue and during surgery in this area. PMID- 6283928 TI - [Anaesthetic problems with glomus tumors of the head and neck (author's transl)]. AB - Anaesthesia and Postanaesthetic Course of Three Cases of Glomus Jugulare Tumor Were Studied. Pathophysiology and clinical features of this very rare tumor is discussed on a brief review of literature. After all localisation and dimension of this tumor can cause serious complications during operation and postoperative period. It must be considered that a safe operative and postoperative period is implied by carefull preoperative diagnostic management, special anaesthetic technics like controlled hypotension as well as postoperative intensive care. PMID- 6283929 TI - Ethylene glycol poisoning. A case report. AB - A case of self-poisoning with ethylene glycol is presented. The metabolic upset induced by ingestion of this substance is discussed and the principles underlying treatment with ethyl alcohol, sodium bicarbonate and renal dialysis are outlined. The practical problems experienced with this therapy are detailed. The need for immediate instigation of treatment and for intensive care are emphasised. PMID- 6283930 TI - A simple and rapid method for purification of profilin from bovine thyroid. PMID- 6283931 TI - Separation of angiotensins and assay of angiotensin-generating enzymes by high performance liquid chromatography. PMID- 6283932 TI - An accurate, quantitative assay for collagenase activity based on the synergistic hydrolysis of collagen. PMID- 6283933 TI - Specific detection of inactive enzyme protein after polyacrylamide gel electrophoresis by a new enzyme-immunoassay method using unspecific antiserum and partially purified active enzyme: application to rat liver phosphodiesterase I. PMID- 6283935 TI - A sensitive fluorometric assay for the determination of DNA. PMID- 6283934 TI - Iac operon operator DNA: isolation and trimming for NMR spectroscopy. PMID- 6283936 TI - A rapid and mild procedure for the isolation of DNA from mammalian cells. PMID- 6283937 TI - Microsequence analysis of peptides and proteins. I. Preparation of samples by reverse-phase liquid chromatography. PMID- 6283938 TI - Characterization of vertebrate collagenase activity by high-performance liquid chromatography using a synthetic substrate. PMID- 6283939 TI - N-[1-13C]acetylimidazole as a reagent for tyrosyl modification in protein NMR studies: acetylation of cytochrome c. PMID- 6283941 TI - Advantages of ferric ferricyanide to detect certain peptides and proteins on chromatograms. PMID- 6283940 TI - An in vitro model to study adipose differentiation in serum-free medium. PMID- 6283942 TI - Rapid and simple measurement of ATP-sulfurylase activity in crude plant extracts using an ATP meter for bioluminescence determination. PMID- 6283943 TI - Modification of the C-terminal octapeptide of cholecystokinin with a high specific-activity iodinated imidoester: preparation, characterization, and binding to isolated pancreatic acinar cells. PMID- 6283944 TI - Identification of enzymatically produced peptide fragments by liquid chromatography and mass spectrometry. PMID- 6283945 TI - Investigation of electron transfer between platinum and large biological molecules by thin-layer spectroelectrochemistry. PMID- 6283946 TI - Determination of the non-starch polysaccharides in plant foods by gas-liquid chromatography of constituent sugars as alditol acetates. PMID- 6283947 TI - Differential sensitivity of fast and slow fibers in mammalian nerve. II. Margin of safety for nerve transmission. AB - In a previous study it was found that concentrations of local anesthetics required to block large, fast-conducting nerve fibers were lower than those required to block small, slow-conducting fibers. The present study was instituted to evaluate the margin of safety for transmission in large versus small nerve fibers, the margin of safety being defined as the ratio between the magnitude of the action potential and the magnitude of the critical membrane potential. The effect of reducing the sodium-activating current, which reduces the magnitude of the action potential by sodium deficient solutions and tetrodotoxin application to the desheathed rabbit vagus nerve trunk (in vitro), was examined. Anode block (another method of reducing sodium current) is also discussed. In all instances, the margin of safety for transmission was greater in small, slow fibers than in large, fast fibers. The variations seen in nerve response to tetrodotoxin application are explained by the presence of nerve fiber diffusion barriers; the large fibers show more diffusion protection than the small fibers. Onset, duration, and intensity of differential nerve blockade by drugs reflect a balance between diffusion barriers and axon membrane sensitivity. PMID- 6283949 TI - Xanthine oxidase-induced lung injury inhibits removal of 5-hydroxytryptamine from the pulmonary circulation. AB - We were interested in determining the effect of lung injury initiated by superoxide anions and hydroxyl radicals on removal of 5-hydroxytryptamine (5-HT) and phenylethylamine by the isolated perfused lung. The rate of removal and percentage of removal of these bioamines was determined before and after lung injury initiated by perfusion of the lung with hypoxanthine (HX) and xanthine oxidase (XO) or xanthine oxidase alone for 10 or 30 minutes; free radicals are generated by such treatment. Because of variation in removal of bioamines among lungs of different animals, the effects of lung injury on bioamine removal were determined by calculating the percentage of inhibition of removal using data from the control and test period for each lung. Perfusion of the lung with HX/XO or XO for 10 or 30 minutes significantly inhibited 5-HT removal by 39.5% and 63.3%, respectively. In contrast, only perfusion of the lung for 30 minutes with HX/XO produced inhibition of phenylethylamine uptake (by 54.8%). As uptake of 5-HT is the rate-limiting step in 5-HT removal, these data demonstrate dose (time) related depression of active 5-HT uptake by free radicals generated in vitro. The rate-limiting step of phenylethylamine uptake, metabolism by monoamine oxidase, is inhibited only by severe lung injury. PMID- 6283948 TI - Differential sensitivity of fast and slow fibers in mammalian nerve. III. Effect of etidocaine and bupivacaine on fast/slow fibers. AB - Etidocaine and bupivacaine are long acting local anesthetics with contrasting effects on motor and sensory function. The effect of these drugs on fast conducting (large, motor) and slow-conducting nerve fibers (small, pain) in the isolated rabbit vagus nerve was examined. Both drugs had an equivalent effect on slow fibers. Etidocaine had a short latency and bupivacaine a prolonged latency of effect on fast fibers. During this long latency of effect by bupivacaine on fast fibers, only the slow fibers were blocked. This period of differential effect on fast and slow fibers is believed to be the explanation for the early effect of bupivacaine on pain fibers followed by a later block of motor function. This difference is believed to be due to the lower lipid solubility solubility and greater ionization of bupivacaine, which impedes diffusion across the permeability barriers present in fast-conducting A fiber. PMID- 6283950 TI - Prolonged exposure to nitrous oxide decreased opiate receptor density in rat brainstem. AB - Groups of rats were exposed to air or 80 per cent nitrous oxide for 30 min or 18 h, following which the brainstem opiate receptor density and the apparent affinity of these receptors to the radiolabeled agonist, 3H-dihydromorphine, were assayed, Thirty-minute exposure to nitrous oxide did not change opiate receptor characteristics, immediately or 17.5 h later. However, prolonged exposure to nitrous oxide (18 h) decreased the brainstem opiate receptor density approximately 20 per cent, without a change in apparent receptor affinity. These results support the view that nitrous-oxide-induced analgesia results from release of endogenous opiate-like substances. Continued presence of these substances in turn results in a decrease in opiate receptor density and may account for the development of tolerance to the analgesic action of nitrous oxide. PMID- 6283951 TI - Pyrimidine-specific 5' nucleotidase activity in bovine erythrocytes: effect of phlebotomy and lead poisoning. AB - Erythrocyte pyrimidine-specific 5' nucleotidase (PY5'N) (E.C. 3.1.3.5) was measured in healthy, anemic, and lead-poisoned calves to determine whether low activity of PY5'N is associated with the propensity of cattle to develop basophilic stippling of erythrocytes. Low activity of PY5'N has been associated with basophilic stippling of erythrocytes in persons with inherited hemolytic anemia and with lead poisoning. A radiometric technique, using [14C]cytidine monophosphate as the substrate, was used to measured PY5'N activity. The erythrocytes from 4 healthy calves had much lower activity (mean of 7.1 +/- 1.6 nmols of [14C]cytidine monophosphate hydrolyzed/min/g of hemoglobin) than has been reported for human erythrocytes. The pH response curve of bovine PY5'N was similar to that of the human enzyme, with maximal activity around pH 7. Experimental hemorrhagic anemia in these calves increased PY5'N activity 6-to 7 fold, with peak activity occurring concomitantly with maximum reticulocytosis. Two of the calves were then given lead per os, and the PY5'N activity decreased within 24 hours to base-line values. In the 2 other calves not given lead, the PY5'N activity declined slowly, but did not reach base-line values after 14 days. PMID- 6283952 TI - DNA of bovine herpesvirus type 1 in the trigeminal ganglia of latently infected calves. AB - Twelve calves infected with bovine herpesvirus type 1 (BHV-1) were killed when in a latent state of infection. Latency was verified 30 days after virus inoculation of the calves by seroconversion, absence of virus shedding, and in 2 calves, by recrudescence of the infection after they were treated with dexamethasone. By in situ hybridization techniques and autoradiography, DNA of BHV-1 was detected in 13 of 23 trigeminal ganglia of latently infected calves. Viral DNA was restricted to the nucleus of nerve cells. Single neurons harboring BHV-1 DNA were observed in 4.9% of the sections (n = 325) of the trigeminal ganglia. The results obtained correspond to those known from herpes simplex virus infections in mice. The implications for the virus-host relationship are discussed. PMID- 6283953 TI - Applications of peroxidase-antiperoxidase staining technique for detection of transmissible gastroenteritis virus in pigs. AB - The peroxidase-antiperoxidase (PAP) staining technique was used for the detection of transmissible gastroenteritis virus (TGEV) in small intestines of TGEV infected 8-week-old pigs and in infected McClurkin pig testicle cells by means of light microscopy. The specific-positive reaction was characterized by the presence of many brown granules of various sizes in the cytoplasm of infected cells. Nonspecific granules caused by endogenous peroxidases in the cytoplasm of eosinophils stained by PAP were darker, larger, more round, and more uniform in size than were specific granules. Acetone fixation was superior to fixation with periodate-lysine-paraformaldehyde or 10% formalin. Our results indicate that the PAP staining technique is a sensitive, specific technique for detection of TGEV in the small intestines of pigs. PMID- 6283954 TI - Effects on ovine fetuses of exposure to ovine progressive pneumonia virus. AB - Fetuses of 20 pregnant ewes at 4 gestational periods (45, 55, 85, and 100 days) were inoculated with ovine progressive pneumonia virus. Fourteen of 16 fetuses exposed to virus before gestational day 80 were either resorbed or expelled, whereas 10 of 15 fetuses exposed to virus after day 80 were normal at birth. Three of the 9 expelled fetuses and 1 of 2 newborn lambs had accumulations of lymphoid cells in the lungs. Virus was readily isolated from the tissues of expelled fetuses and newborn lambs. Lambs did not have precipitating antibody to the virus at birth, but 3 to 5 lambs had specific antiviral antibody at 18 months of age. PMID- 6283955 TI - Border disease virus isolation from postpartum ewes. AB - The prevalence of border disease virus in a flock of Targhee sheep in southern Idaho was determined by serologic and virologic techniques. Of 249 ewes, 73 (29%) were seropositive to the antigenically related togavirus, bovine seropositive to the antigenically related togavirus, bovine viral diarrhea virus (BVDV). Concurrent testing of sera from 337 neonatal lambs indicated that 172 (51%) were seropositive for BVDV. It was observed that a high percentage (35%) of the BVDV seropositive lambs were derived from seronegative ewes. Immunotolerance of the ewe was considered as the explanation for the lack of serologic relationship between seropositive lambs and their seronegative dams. Eleven cytopathogenic viruses were isolated from nasal and vaginal specimens obtained from 80 clinically healthy ewes within 24 hours of parturition. The viral isolates were demonstrated to be antigenically related to BVDV by direct immunofluorescence. Six of the 11 isolates were obtained from BVDV-seronegative ewes. PMID- 6283956 TI - Adrenocortical suppression in the dog given a single intramuscular dose of prednisone or triamcinolone acetonide. AB - Adrenocortical function was assessed in dogs given a single intramuscular dose of either prednisone or triamcinolone acetonide (TCA; or saline solution to controls) to determine the duration of adrenocortical suppression caused by 2 commonly used glucocorticoids. The glucocorticoids were administered at recommended therapeutic doses; therefore, dogs given prednisone received a greater amount of glucocorticoid activity than did in dogs given TCA. Basal and ACTH-stimulated plasma cortisol concentrations, as determined by radioimmunoassay, were obtained once a week. Total intravascular eosinophil concentration and skin responses to intradermally injected histamine phosphate were quantitated. Dogs given TCA showed suppressed basal and ACTH-stimulated plasma cortisol concentrations 1 week after injection; the latter change persisted 2 weeks after injection. Adrenocortical function in 1 of 4 dogs given TCA remained suppressed for 4 weeks. In contrast, prednisone did not significantly alter adrenocortical function. Although intravascular eosinophil concentrations did not vary among groups, skin responses to intradermally injected histamine phosphate were reduced 6 days after prednisone and TCA were given. PMID- 6283957 TI - Intestinal, pulmonary, and serum antibody responses of feeder pigs exposed to transmissible gastroenteritis virus by the oral and the oral-intranasal routes in inoculation. AB - Specific antibodies against transmissible gastroenteritis (TGE) virus produced as a result of oral and oral-intranasal viral exposure of feeder pigs were isolated by molecular sieve column chromatography of lung wash, intestinal fluid, and serum samples. Selected fractions were analyzed for neutralizing antibody activity. Oral inoculation gave a predominantly immunoglobulin (Ig) G antibody response in lung wash and serum, as compared with the response in intestinal fluid in which neutralizing antibody activity was found in the IgA-containing fractions. Oral-intranasal inoculation produced a predominantly IgA antibody response in intestinal fluid. Lung wash contained IgA and IgG antibody, and serum antibody consisted primarily of IgG class of Ig. Neutralizing antibody (IgA, IgG, IgM) from the lung wash, intestinal fluid, and serum samples persisted beyond 56 days after oral and oral-intranasal viral exposures. The virus-shedding period in feces was 1 to 7 days after oral or oral-intranasal inoculation of feeder swine with virulent TGE virus. However, TGE virus was reisolated from lung and intestinal tissue for as long as 42 and 56 days, respectively. PMID- 6283958 TI - Experimental transmission of pseudorabies virus in swine by embryo transfer. PMID- 6283959 TI - Horizontal transmission of pseudorabies virus in cattle. AB - Pseudorabies virus (PRV) was not transmitted horizontally from 3 PRV-infected calves to 2 contact control calves during 4 days of comingling in experiment 1. Although these contact control calves developed clinical signs of pseudorabies when infected intranasally with PRV in experiment 2, they did not transmit PRV to a second pair of contact control calves. However, 1 of 2 pigs comingled with these 4 calves seroconverted. During both experiments, moderate amounts (10(2) to 10(5) TCID50) of PRV were present in the nasal secretions of the infected calves during the contact periods. All infected calves traumatized their nares or periorbital tissue. Infected calves developed a nonsuppurative meningoencephalitis mainly involving the brain stem. Four of the 5 infected calves had nonsuppurative ganglioneuritis and acute lymphoid necrosis of germinal centers. Virus could not be recovered from nasal and tonsillar swab samples from contact-control calves and pigs. PMID- 6283960 TI - Recrudescence of bovine herpesvirus-5 in experimentally infected calves. AB - A latent infection of bovine herpesvirus-5 (BHV-5) was established in 4 calves. These calves, plus 2 controls, were given dexamethasone (DM) to reactivate the latent virus. The 4 principal calves developed antibodies to BHV-5 by postinoculation day (PID) 21. Antibody titers increased until PID 42 before decreasing to low levels of PID 75. After the first DM treatment (started on PID 76), an anamnestic antibody response was demonstrated in the 4 principal calves. Calves, 2, 3, and 4 were euthanatized and necropsied at PID 121, and their antibody titers were again decreasing. The virus BHV-5 was not isolated from the tissues by conventional techniques of viral isolation but was isolated from the trigeminal ganglion and spinal cord of calf 3 by explantation techniques. The BHV 5 was isolated, using conventional viral isolation techniques, from a nasal swab sample of calf 1 on PID 91 (15 days after the first DM treatment) and from the thoracic lymph node 6 days after the start of a 2nd DM treatment. Seemingly, BHV 5 may be latently harbored in the nerve tissues or calves and this virus may be reactivated from the upper respiratory tract following subsequent DM treatment. PMID- 6283961 TI - Detection of anti-rotavirus antibody-producing cells in paraffin-embedded tissue sections. AB - An assay to detect anti-rotavirus antibody-producing cells was developed in rabbits. Sections of alcohol-fixed, paraffin-embedded tissues were used to make an immunofluorescent "sandwich" consisting of specific antibody-producing cells, a concentrated suspension of bovine rotavirus, and rhodamine-conjugated anti rotavirus serum. Rhodamine-stained, anti-rotavirus antibody-producing cells could be simultaneously stained by a fluorescein-conjugated antiserum reactive to rabbit immunoglobulin. PMID- 6283962 TI - Chemotactic requirements of bovine leukocytes. AB - A system was described for studying bovine leukotaxis. Chemotaxis was readily observed toward bovine serum activated by zymosan of agarose. However, bacterial culture filtrates and peptides, which were potent chemotaxins for leukocytes from other species, failed to affect bovine leukotaxis. Using conditions suitable for studying binding to leukocytes from other species, specific binding of a radiolabeled chemotactic peptide to bovine leukocytes was not apparent. These data indicate that bovine leukocytes have chemotactic requirements distinct from those of other species. PMID- 6283963 TI - Vaccination of cattle against pseudorabies (Aujeszky's disease) with homologous virus (herpes suis) and heterologous virus (herpes bovis 1). AB - Study was made to determine whether vaccination of cattle against pseudorabies (PR; Aujeszky's disease) affords protection upon subsequent intranasal challenge exposure with virulent virus. Vaccinations were performed with a commercially available oil-adjuvant PR virus vaccine, in some cases supplemented with A1(OH)3 (given subcutaneously), with the attenuated NIA4 strain of PR virus (given intranasally), and with a commercially available temperature-sensitive mutant of bovine herpesvirus-1 (infectious bovine rhinotracheitis [IBR] virus) given intranasally. Challenge exposure was performed intranasally with 10(3) median lethal doses (LD50) of the virulent PR virus. In earlier experiments, it was found that LD50 median tissue culture infective doses of this virulent PR virus in primary pig kidney cells represented about 1 LD50 for cattle. This LD50 was practically no different for cattle whether or not they had antibodies against PR virus before exposure to virulent PR virus. In the seropositive animals, antibodies were present presumably as a consequence of a previous IBR infection in the field. Vaccination intranasally with NIA4 or IBR virus did not result in serologic response or protection against challenge exposure. The inactivated virus vaccine induced a good serologic reaction, but there was poor protection against challenge exposure. Consequently, vaccination of cattle at risk of exposure to virulent PR virus cannot be advised. In the present experiments, initiation of infection with virulent PR virus in cattle always resulted in disease and death. PMID- 6283964 TI - Failure of swine influenza virus to cause transplacental infection of porcine fetuses. PMID- 6283965 TI - Experimental transplacental infection of pregnant dogs with canine herpesvirus. AB - Fetal infection was established in 28 of 33 pups obtained from 7 pregnant dogs (47th to 53rd day of gestation) inoculated IV with canine herpesvirus (CHV). The pathogenesis of transplacental CHV infection was investigated by using pathologic examination, immunofluorescence microscopy, and viral assay. Infection with the virus was recognized in 13 of 18 neonatal pups obtained from 4 bitches by spontaneous delivery at full term. Eleven pups died within a week of birth and all had characteristic lesions of CHV infection. Thirteen live-born and 2 stillborn pups derived from 3 bitches by cesarean section died within 9 days of birth. Characteristic lesions of CHV infection were observed in various organs, such as liver, spleen, kidney, and lung. Fourteen of the fetal placentas had variable degree of necrotizing lesions. The lesions were characterized by degenerative changes and focal necrosis in the placental labyrinth. Lesions were present in the walls of allantoic blood vessels. Intranuclear inclusions were evident in degenerating trophoblast cells and in the cells of maternal and fetal blood vessels. Infective virus and viral antigen were detected in the various organs of pups and in the trophoblastic cells of junctional zone. Virus also was found in the walls of maternal and fetal blood vessels of the placental tissues. The results strongly indicated that transplacental transmission of CHV may occur in pregnant dogs at a late stage of gestation. PMID- 6283966 TI - Detection of antibody in Aleutian disease of mink: comparison of enzyme-linked immunosorbent assay and counterimmunoelectrophoresis. AB - Experiments were undertaken to investigate the potential of the enzyme-linked immunosorbent assay (ELISA) as a screening test for the diagnosis of the 2 known naturally occurring forms of Aleutian disease of mink. Anti-Aleutian disease virus (ADV) antibody activity was not detectable in the sera of mink with nonprogressive Aleutian disease despite the demonstration of antibody by counterimmunoelectrophoresis (CIEP) in the same sera. Anti-ADV antibody was detectable in 93% of sera from mink at various stages of experimentally induced progressive Aleutian disease. False-negative reactions occurred in sera which demonstrated high anti-ADV antibody titers by CIEP. As a consequence of the high prevalence of false-negative reactions, the ELISA was not considered to be an effective screening test. However, using CIEP as an indicator of ADV infection, the ELISA may be useful in differentiating mink with nonprogressive Aleutian disease from mink with progressive Aleutian disease. PMID- 6283967 TI - Bluetongue infections in Louisiana cattle. AB - Bluetongue virus (BTV) serotype 17 was isolated from cattle with clinical signs of bluetongue disease during 1978 and 1979 epizootics. Bovine sera from 6 herds located in an epizootic region were examined in 1979 for antibodies, using an immunodiffusion (ID) test. Of 300 sera, 164 (54.7%) were seropositive. Sera from statewide surveys of Louisiana cattle in July to August 1980 and December 1980 to January 1981 were tested for BTV antibodies, using the ID test. Fifty-eight of 70 herds (82.9%) and 164 of 597 (27.5%) individual cattle tested in July to August 1980 were seropositive. Fifty-four of 63 (85.7%) herds and 170 of 600 (28.3%) individual cattle tested in December 1980 to January 1981 were seropositive. Significant differences (P less than 0.01) were found in the seropositive rates between the various geographic regions of the state during each survey. Adult breeding-age cattle in 3 sentinel herds were tested for BTV antibodies beginning in 1976 and continuing through January 1981. During this interval, the seropositive rate in 2 of 3 herds was increased. Also, individual cattle in all 3 of these herds converted from seronegative to seropositive, indicating exposure during a particular interval for each herd. The age distribution of seropositive cattle in a dairy indicated that 2-year-old cattle had a seropositive rate comparable with that of older animals in the herd, suggesting that the 2-year-old animals had been exposed to a BTV before they entered the breeding herd. PMID- 6283968 TI - Bacterial cultures of the lower respiratory tract in normal volunteers with and without experimental rhinovirus infection using a plugged double catheter system. AB - To test a method of obtaining specimens for microbiologic culture from the lower respiratory tract, we bronchoscoped 25 otherwise normal subjects with and without experimental rhinovirus infections and collected specimens for bacterial culture with a brush housed in a plugged double catheter apparatus. Fifty-two specimens of the lower respiratory tract were obtained from the 25 subjects; nasopharyngeal swabs were also obtained from each subject prior to bronchoscopy. Specimens obtained from the lower respiratory tract were positive for bacteria in 21 (84%) of 25 subjects and at 38 (73%) of 52 sites. The frequency of obtaining positive bacterial cultures was similar in volunteers with (69%) or without (90%) rhinovirus infection. Oropharyngeal contamination of bronchial specimens was minimized by administering atropine and by bronchoscoping subjects in the supine or Trendelenburg position. These data indicate that the plugged double catheter brush system does not consistently yield specimens that are free of bacterial contamination from the oropharynx. PMID- 6283969 TI - Ketoconazole therapy of murine cryptococcal meningitis. AB - Currently, even optimal therapy of cryptococcal meningitis is associated with appreciable mortality, drug toxicity, and prolonged hospitalization. Ketoconazole, a new oral imidazole, has therefore been evaluated in a murine model of cryptococcosis. Cryptococcal meningitis was induced in BALB/c mice by intracranial injection of Cryptococcus neoformans. The mice developed infection characterized by diffuse meningitis and extracerebral dissemination. Oral ketoconazole therapy prolonged survival of infected mice, but most mice ultimately succumbed to infection. Ketoconazole dramatically reduced the cryptococcal counts in the liver, but had minimal effect on counts in the brain. Paralleling these results were high concentrations of ketoconazole found in lung, spleen, and heart muscle and lower concentrations (2 to 5 micrograms/ml) found in the brain. The detection of biologically active drug in the brain, and the prolongation of survival afforded by ketoconazole, suggest a potential role for this agent in the theory of cryptococcal meningitis. PMID- 6283970 TI - Ketoconazole therapy for systemic fungal infections: inadequacy of standard dosage regimens. AB - We treated 29 patients with ketoconazole for systemic mycoses. Twenty-two had coccidioidomycosis, 5 had histoplasmosis, and 2 had sporotrichosis. Of the 25 patients who received 200 mg/day, 16% improved on that dose. Of 17 patients who did not improve on 200 mg/day, 71% responded after the dose was increased to 400 or 800 mg/day. Treatment periods ranged from 21 days to more than 600 days. Five patients relapsed. In 2, ketoconazole had been reduced in dose or discontinued. In 3, the drug was still being given. Ketoconazole is effective in suppressing disease in various mycoses. However, patients may require high doses for prolonged periods to achieve maximal benefit. PMID- 6283971 TI - Bronchioloalveolar carcinomas: a heterogenous group. PMID- 6283972 TI - Biliary and gastric decompression by a transhepatic tube technique. PMID- 6283974 TI - Opportunistic infections and immune deficiency in homosexual men. AB - A syndrome of opportunistic infections and acquired immune deficiency occurred among four previously healthy homosexual men. Fever, leukopenia, and diminished delayed hypersensitivity were accompanied by various degrees of proctitis, perianal ulcerations, and lymphadenopathy. The infectious agents included Pneumocystis carinii, Cryptococcus neoformans, Candida albicans, herpes simplex virus, and cytomegalovirus. The immune deficiency was characterized as a persistent and profound selective decrease in the function as well as number of T lymphocytes of the helper/inducer subset and a possible activation of the suppressor/cytotoxic subset. Three patients died despite aggressive anti infective therapy. PMID- 6283973 TI - Disseminated Kaposi's sarcoma in homosexual men. AB - Nineteen cases from an epidemic of disseminated Kaposi's sarcoma in homosexual men were studied by clinical, virologic, immunologic, and genetic methods. The patients were all male homosexuals ranging in age from 29 to 52 years, with histories of multiple sexually transmitted diseases and exposure to both prescription and recreational drugs. Sites of disease included skin (16 of 19 patients), lymph nodes (13 patients), gastrointestinal tract (12 patients), spleen (three patients), and lung (one patient). Most patients had elevated levels of serum immunoglobins, positive antibody titers to hepatitis A and B virus, cytomegalovirus and Epstein-Barr virus, and impairment of cell-mediated immunologic reactions. The frequency of HLA-DR5 in these patients was significantly elevated. Two of the 19 patients died. Although the precise cause of this epidemic is unknown, it is likely that a genetic predisposition, an acquired immunoregulatory defect, and one or more infectious agents and drugs may be involved. PMID- 6283975 TI - An outbreak of Pneumocystis carinii pneumonia in homosexual men. AB - Pneumocystis carinii pneumonia has rarely been reported in previously healthy persons over the age of 6 months. Five cases of P. carinii pneumonia in adult homosexual men, confirmed by biopsy results, are reported. All five patients were seropositive when tested for antibodies to cytomegalovirus and four had evidence of active concurrent cytomegalovirus infections. Kaposi's sarcoma was shown in two of the patients and one had possible Pneumocystis infection of the central nervous system as well as P. carinii pneumonia. Three patients had second episodes of Pneumocystis pneumonia. Four of the five patients have died. Past or concurrent cytomegalovirus infection and homosexuality were the only common epidemiologic features in all five patients. PMID- 6283976 TI - Infectious mononucleosis in an adult progressing to fatal immunoblastic lymphoma. AB - We report a case of infectious mononucleosis progressing to fatal immunoblastic lymphoma. The patient, a 44-year-old man who may have had an immunoregulatory defect, failed to have an appropriate T-cell response to his Epstein-Barr (EB) viral infection. His active EB viral infection was manifest by seroconversion of IgM-viral capsid antibody and a greater than four fold rise in IgG-viral capsid antibody. Also, he transmitted his EB viral infection to his wife who became ill 1 month after his death. Clinically the patient's illness was characterized by waxing and waning lymphadenopathy, persistent fever, diarrhea (similar to that associated with cholera), a coagulopathy, and gastrointestinal bleeding. The patient had pathologic findings of a diffuse immunoblastic lymphoma involving lymph nodes, small bowel, liver, pancreas, kidneys, lungs, and bone marrow. Immunologic cell markers showed the tumor to be polyclonal. PMID- 6283977 TI - Epidemiology of Norwalk gastroenteritis and the role of Norwalk virus in outbreaks of acute nonbacterial gastroenteritis. AB - Outbreaks of Norwalk gastroenteritis, which may involve persons of all ages, occur during all seasons and in various locations. Waterborne, foodborne, and person-to-person modes of transmission have been described, and secondary person to-person transmission is common. Outbreaks generally end in about 1 week; longer outbreaks occur only when new groups of susceptible persons are introduced, usually in the setting of a persistent common source of infection. The illness is generally mild and characterized by nausea, vomiting, diarrhea, and abdominal cramps. Vomiting is the predominant symptom among children, whereas diarrhea is commoner among adults. Forty-two percent of 74 outbreaks of acute nonbacterial gastroenteritis investigated by the Centers for Disease Control from 1976 to 1980 were attributed to the Norwalk virus. The rest resembled Norwalk outbreaks clinically and epidemiologically and were probably caused by 27-nm viral agents similar to the Norwalk virus. PMID- 6283978 TI - Small-cell lung cancer and limbic encephalitis. PMID- 6283979 TI - Cortisol, valproic acid, and ACTH suppression. PMID- 6283980 TI - Hepatic dysfunction and ketoconazole therapy. PMID- 6283981 TI - Toxic shock syndrome: case-control studies at the Centers for Disease Control. PMID- 6283983 TI - Cushing's syndrome and the changing times. PMID- 6283982 TI - Cytomegalovirus immune plasma in bone marrow transplant recipients. AB - The effects of passive immunization on cytomegalovirus infection and interstitial pneumonia in marrow transplants were evaluated in a randomized, controlled trial. Twenty-four patients received cytomegalovirus immune plasma before and after transplantation, and 24 patients were controls. Although the incidence of cytomegalovirus infection was similar in the control and plasma groups, symptomatic infection (12 of 24 versus five of 24, p = 0.07) and interstitial pneumonia (11 of 24 versus five of 24, p = 0.12) occurred less frequently in the group receiving plasma. Cytomegalovirus infection occurred in 11 of 13 recipients of leukocyte transfusions and in 16 of 35 patients not given leukocyte transfusions (p = 0.02). Among patients not given leukocyte transfusions, the incidence of cytomegalovirus infection was similar in the control and plasma groups, but symptomatic infection (eight of 18 versus one of 17, p = 0.03) and interstitial pneumonia (nine of 18 versus one of 17, p = 0.01) were significantly less in the group receiving plasma. These results suggest that passive immunization modifies cytomegalovirus infection in humans and prevents interstitial pneumonia in marrow transplants especially when leukocyte transfusions are not used. PMID- 6283984 TI - Benign pulmonary nodule and small-cell cancer. PMID- 6283985 TI - DNA modification and repair in the experimental induction of nervous system tumors by chemical carcinogens. PMID- 6283986 TI - Photoperiod and regulation of gonadotropin receptors. PMID- 6283987 TI - Biochemical properties of the testicular follitropin-receptor system. AB - The interaction of FSH with membrane receptors from rat and calf testis has been studied in some detail. The FSH receptor has been solubilized through use of the nonionic detergent Triton X-100 and highly purified by affinity chromatography on Affigel-10 coupled to ovine FSH. Hormone binding activity of the solubilized receptor has been preserved for extended periods through use of the structure stabilizing agent glycerol. Other components of the FSH testes receptor system including the guanyl nucleotide binding protein and adenylate cyclase have been solubilized by nonionic detergents and also found to be stabilized by glycerol. FSH binding activity has been observed in testes cytosol and represents a putative class of receptors prepared from testes in the absence of detergent. The concentration of this buffer-soluble component decreased with age and increased concomitantly with loss of membrane receptors consequent to their down-regulation after administration of exogenous FSH. Phospholipids seem involved in the interaction of FSH with membrane-bound, detergent-solubilized, and buffer-soluble FSH binding activity. Phospholipids may maintain or stabilize a particular receptor conformation necessary for interaction with the hormone. A specific role for GTP seems indicated in regulation of FSH-stimulated adenylate cyclase activity in immature rat testis. Follitropin binding to testes receptor appears modulated by a variety of factors present in serum, testes extracts, follicular fluid, and seminal plasma, which are poorly understood at present. Inhibition of FSH binding by seminal plasma best-fit by a model proposing two hormone binding sites per receptor molecule, where binding to one site decreases the affinity of the other site for FSH. As a result of studies in this and other laboratories, the molecular endocrinology of FSH interaction with testis receptors is becoming increasingly understood. PMID- 6283988 TI - Receptor-mediated uptake and degradation of human chorionic gonadotropin: fate of the hormone subunits. PMID- 6283989 TI - LH receptors and steroidogenesis in distinct populations of Leydig cells. PMID- 6283990 TI - Purification of rat Leydig cells: functional and morphological evaluation. PMID- 6283991 TI - Gonadotropin-induced desensitization of Leydig cells in vivo and in vitro: estrogen action in the testis. PMID- 6283992 TI - In vivo and in vitro response of Leydig cells to acute stimulation by hCG. PMID- 6283993 TI - Direct effects of gonadotropin releasing hormone on testicular Leydig cell functions. PMID- 6283994 TI - The secretion of androgen-binding protein and other proteins by rat Sertoli cells in culture: a structural and electrophoretic study. PMID- 6283995 TI - Sperm-egg binding events during sea urchin fertilization. PMID- 6283996 TI - Regulation of the cytoskeleton by Ca2+-calmodulin and cAMP. PMID- 6283997 TI - Critical issues in the use of vasoactive substances to assess lund microvascular injury. PMID- 6283998 TI - Metabolism of vasoactive peptides by membrane-enriched fractions from human lung tissue, pulmonary arteries, and endothelial cells. PMID- 6283999 TI - System diseases and visual evoked potential diagnosis in neurology: changes due to synaptic malfunction. PMID- 6284001 TI - Analysis of click-evoked brainstem auditory electric potentials using high-pass noise masking and its clinical application. PMID- 6284000 TI - Somatosensory evoked potentials in man: subcortical and cortical components and their neural basis. PMID- 6284002 TI - Developmental dependencies of the human brainstem auditory evoked response. PMID- 6284003 TI - Would an evaluation of binocularity using algebraic interaction between the two eyes be modified using the pseudorandom binary sequence method? PMID- 6284004 TI - Chronotopographical study of the pattern-evoked response and binocular summation. PMID- 6284005 TI - Natural stimuli evoking somatosensory potentials. PMID- 6284007 TI - Hard Retinal exudates and visual loss due to papilledema. AB - Bilateral papilledema developed in a patient with a cystic, grade 3 astrocytoma of the right frontal lobe. Despite successful neurosurgical treatment, 60Co radiotherapy, and oral corticosteroid therapy, progressive visual loss occurred. At examination one year later, visual activity was 20/200 and 20/70, and extensive lipid exudates in the peripapillary retina and central macula of each eye were noted. Retinal lipid exudates rarely complicate the course of surviving patients who had papilledema from intracranial tumor; physicians involved in the multispecialty care of such patients should be aware of the possible ocular residuals of persistent papilledema in an otherwise successfully treated patient. PMID- 6284006 TI - Ceruloplasmin: an acute phase reactant that scavenges oxygen-derived free radicals. AB - Superoxide anion radicals have been implicated recently as mediators of inflammation and tissue injury. Protection from superoxide anion radicals is provided primarily by a copper-containing, intracellular enzyme (superoxide dismutase) (SOD) that catalyzes the dismutation of superoxide to hydrogen peroxide and oxygen. We have found that the action of cytoplasmic SOD to scavenge superoxide and thereby to inhibit superoxide-mediated reactions can be mimicked by the copper-containing plasma protein and acute-phase reactant, ceruloplasmin. Ceruloplasmin, at concentrations present in normal plasma, inhibited reduction of both cytochrome c and nitroblue tetrazolium (NBT) mediated by the aerobic action of xanthine oxidase on hypoxanthine (a superoxide-generating system). Ceruloplasmin neither inhibited formation of uric acid by xanthine oxidase nor accelerated autooxidation of cytochrome c. Furthermore, in an experimental system in which contact between ceruloplasmin and indicator was prevented by a relatively impermeable lipid membrane barrier, ceruloplasmin inhibited reduction of NBT trapped within liposomes exposed to xanthine oxidase and hypoxanthine. Ceruloplasmin also inhibited reduction of cytochrome c and NBT mediated by the aerobic action of xanthine oxidase on acetaldehyde (another superoxide-generating system) and mimicked the activity of purified human erythrocyte SOD by inhibiting photoreduction of NBT and by accelerating aerobic photooxidation of dianisidine. Ceruloplasmin could be separated from purified human erythrocyte SOD by electrophoresis on alkaline 12% polyacrylamide gels and identified by its superoxide-scavenging activity. These results suggest that ceruloplasmin may function as a circulating scavenger of oxygen-derived free radicals. PMID- 6284008 TI - Granular cell tumor of the orbit: a case report including electron microscopic observations. PMID- 6284009 TI - Malignant mixed tumor. PMID- 6284010 TI - Mobilization of both breasts for reconstruction of a midline anterior chest wall defect. PMID- 6284011 TI - [Adaptation of an isolate of bovine enteritic coronavirus to serial growth in cell culture and characterization of the virus (author's transl)]. AB - Coronaviruses are known to behave as pathogens in a variety of animal species. Their multiplication in the differentiated enterocytes of the intestinal tract induces a severe diarrheic syndrome which may lead to death. To study these viruses and to progress in the development of prophylactic and diagnostic methods, it is essential to succeed in adapting the agents to serial growth in cell culture. In spite of several attempts adaptation to primary cell cultures could be obtained in the past only for two isolates. The present work describes the methodology which enabled us to succeed in the adaptation of a bovine enteric coronavirus isolated in France to primary calf kidney cells, and reports results on the characteristics of this virus. Adaptation to serial growth was realized at 34 degrees C in the following way: 3 passages each lasting 8 to 10 days followed by 16 passages each lasting 6 days. From the 20th passage on the duration of incubation was reduced to 4 days. Virus growth was checked using an indirect immunofluorescence test. During the first passages only a few dispersed positive cells could be observed but later on small foci of immunofluorescent cells did appear. By the 35th passage infective titers did not exceed 1 x 10(5) TCID50/ml and cytopathic effects were always very discrete. When the susceptibility of different cell-types to this coronavirus strain (G110) was checked we found that only bovine cells and the human HRT18 cell-line were able to replicate the virus. In HRT18 cells titers as high as 5 x 10(7) TCID50/ml could be obtained at the first passage of G110 strain, previously adapted to primary calf kidney cells. As regards the antigenic properties of G110 virus no difference could be established between this strain and the Nebraska isolate (NCDCV) neither by indirect immunofluorescence test nor by indirect neutralization test. The G110 strain agglutinates rat and mouse erythrocytes. The density of purified virions in mouse gradients is 1.19 and the morphology of the particles is characteristic of a coronavirus. PMID- 6284012 TI - [Incidence of rotavirus infection and in combined infection of rotavirus and enteropathogenic Escherichia coli in French calves (author's transl)]. AB - Detection of a rotavirus in faeces of 789 calves developing diarrhoea gave positive results among 48% of the calves. The same investigation extended to 96 apparently healthy animals shows that 12.5% of the faeces contained rotavirus. It appears that all the last ones cannot be considered as healthy controls. Association of rotavirus and E. coli K99+ is found in 5% of sick animals less than 10 days old. PMID- 6284013 TI - The natriuretic hormone. PMID- 6284014 TI - Dietary therapy of hyperlipoproteinemias: new understandings. AB - Diet is the common denominator in the treatment of hyperlipidemia. Calorie and alcohol restriction are often prescribed for hypertriglyceridemic subjects. When these subjects lose weight their serum triglycerides often decrease, secondary to a diminution in hepatic triglyceride secretion. There is also a reduction in insulin resistance leading to an improvement in carbohydrate tolerance. Because some hypertriglyceridemic subjects over-synthesize triglycerides after alcohol ingestion, alcohol restriction is important in the dietary therapy of these patients. Although controversial, the restriction of cholesterol and saturated fat intake is often prescribed for hypercholesterolemic subjects. Recent evidence show (a) As the daily absolute cholesterol intake increases, the % absorbed is decreased but the amount absorbed per kg body weight is increased. (b) Hypercholesterolemic subjects differ from normal subjects in their response to cholesterol and fat intake. (c) A high cholesterol and high saturated fat diet increases the cholesterol concentration in all lipoprotein fractions. A low cholesterol and high polyunsaturated fat diet has the opposite effect. (d) These diets also affect serum apoprotein levels (apo B and apo A-I). It is becoming evident that hyperlipidemic subjects respond differently from normal subjects to dietary changes. For these subjects, at greater risk of developing atherosclerosis, dietary therapy is important. PMID- 6284016 TI - Neuroendocrine small cell carcinomas in miscellaneous primary sites: implications for staging and therapy. PMID- 6284017 TI - Autologous antibodies to meningioma cell surface antigens. AB - Sera of 32 patients with meningioma were tested for reactivity to cell surface antigens of autologous meningioma cells with protein-A assay (PA), immune adherence assay (IA), and anti-C 3-mixed hemadsorption assay (C3-MHA). Antibodies against autologous meningioma could be detected in 6/32 patients by PA, in about half the patients by IA and in almost all patients by C3-MHA with titers ranging from 1:2 to 1:28. Only the serum reactivity detected by C3-MHA was high enough for analysis of the specificity of the reaction by absorption tests. By absorption with a panel of autologous, allogeneic and heterologous cells we were unable to demonstrate a meningioma-specific antigen. Most autologous sera detected oncofetal antigens. Serum reactivity to autologous meningioma showed no correlation to antibodies to SV-40. PMID- 6284018 TI - Binding of the anticancer drug cis-dichlorodiammineplatinum to 3'(2')-guanosine monophosphate and nucleic acids. AB - The UV-absorption spectral characteristics of cis[Pt(NH3)2(GMP)2]-2 ([Pt(NH3)2 (Guanosine-3'(2')-monophosphate)2]-2) (abbreviated Pt (GMP)2) and cis [Pt(NH3)2(Guanine)2]+2 (abbreviated Pt(Gua)2) were studied at acidic, neutral and alkaline pH values. The compound Pt(GMP)2 was formed by reacting GMP with cis Pt(NH3)2Cl2(= cis-platinum) in H2O(pH approximately 6) at 24 degrees C for 7 days in the dark. Hydrolysis of Pt(GMP)2 with HClO4 to remove its sugar-phosphate moiety followed by paper chromatographic separation yielded Pt(Gua)2. Exposure to alkali did not cause any irreversible change in the absorption spectrum of Pt(GMP)2 indicating the lack of imidazole ring fission. Spectral changes for Pt(Gua)2 as compared to control Gua at neutral and alkaline pH values were quantitatively less than those reported for 7-methyl guanine in the literature. Electronic perturbation of the guanine ring system was thus found to be considerably less for platinum than for alkylation of the N7 site. These subtle differences between the platination and alkylation of Gua are of interest in view of the apparent similarities between the biological effects of cis-platinum and bifunctional alkylating agents reported in the literature. PMID- 6284015 TI - Red cell enzymopathies in the newborn. II. Inherited deficiencies of red cell enzymes. PMID- 6284019 TI - Detection of herpes simplex virus mRNA in latently infected trigeminal ganglion neurons by in situ hybridization. AB - Latent infection of the trigeminal ganglion with herpes simplex virus type 2 (HSV 2) was studied in guinea pigs by in situ DNA hybridization. Frozen ganglion sections from animals killed during the period of latent virus infection were studied under nondenaturing conditions. Some sections were treated with deoxyribonuclease (DNase) or ribonuclease (RNase) before incubation with HSV DNA probes. HSV probes consisted of viral DNA nick translated and labeled in vitro with tritiated nucleotides. Bacteriophage lambda DNA, similarly prepared, was used as a control probe. The lambda probe was negative in all situations, including HSV-2-infected monolayer cells in cell culture. HSV-2 probes produced heavy label and, therefore, evidence of hybridization with HSV-2-infected monolayer cells. When HSV-2 probes were incubated with latently infected ganglion sections, hybridization was detected in 71% of guinea pigs and 46% of ganglia. Label was seen only in neurons, and in positive ganglia 0.3 to 5% of neurons were labeled. The amount of label was markedly decreased by pretreatment of ganglion sections with RNase but not DNase, indicating that the DNA probes hybridized to HSV messenger RNA in the latently infected ganglia. PMID- 6284020 TI - [Molecular models of DNA binding with antineoplastic antibiotics specific for certain DNA bases]. PMID- 6284021 TI - [Complete structure of the glycopeptide antibiotic ristomycin A]. PMID- 6284022 TI - [Structure of actinoidins A and B]. AB - The sequence of the amino acid residues in the peptide chain of aglycone was shown on the basis of the data concerning the splitting of actinoidin aglycon and the products of its partial acid hydrolysis by various methods. It was found that the free COOH-group of actinoidin belonged to he glycine residue of the dioxyphenyl nucleus of actinoidinic amino acid. The site of the actinosamine attachment to aglycon was determined. The final structure of actinoidins A and B is suggested. PMID- 6284023 TI - [Spatial configuration of the carbohydrate portion of carminomycins II and III]. AB - A neutral fragment of the carbohydrate part of the molecule was isolated in the form of a cyclic acetal (3.5-dinitrobenzoate) from carminomycins II and III belonging to anthracyclines. The acetal preserved all the asymmetric centers. Methanolysis of the cyclic acetal resulted in formation of aldol dimethyl acetal and propylene glycol (3,5-dinitrobenzoate) with isolated asymmetric centers. On the basis of the optic and spectral properties of these compounds it was found that carminomycins II and III differed in the configuration of 2 asymmetric carbon atoms, i.e. acetal atom and the atom in propylene glycol. They had configurations of R (--) in carminomycin II and S (+) in carminomycin III. The third asymmetric center of the acetal in both antibiotics was the same, i.e. S(+). PMID- 6284024 TI - [Polythiazole-containing peptide antibiotics]. PMID- 6284025 TI - Biosynthesis and biological actions of prostaglandins and thromboxanes. PMID- 6284027 TI - Control of Mucor rouxii adenosine 3':5'-monophosphate phosphodiesterase by phosphorylation--dephosphorylation and proteolysis. PMID- 6284026 TI - The scavenging of superoxide radical by manganous complexes: in vitro. PMID- 6284028 TI - Modified oscillation behavior and decreased D-3-hydroxybutyrate dehydrogenase activity in diabetic rat liver mitochondria. PMID- 6284029 TI - Defect in 3'-phosphoadenosine 5'-phosphosulfate synthesis in brachymorphic mice. I. Characterization of the defect. PMID- 6284030 TI - Defect in 3'-phosphoadenosine 5'-phosphosulfate synthesis in brachymorphic mice. II. Tissue distribution of the defect. PMID- 6284031 TI - Activation of cyclic AMP-dependent protein kinase by epinephrine in proliferating lymphoid cells. PMID- 6284032 TI - Stimulation of cyclic nucleotide phosphodiesterase by products of phosphatidylinositol metabolism catalyzed by phospholipase A2. PMID- 6284033 TI - Adenosine diphosphate-ribosyltransferase activity in permeabilized mouse testicular cells. PMID- 6284034 TI - Rabbit liver fructose-1,6-bisphosphatase: location of an active site lysyl residue in the COOH-terminal fragment generated by a lysosomal proteinase. PMID- 6284035 TI - Influence of mevalonate kinase on studies of the MgATP-dependent inactivator of 3 hydroxy-3-methylglutaryl coenzyme A reductase. PMID- 6284036 TI - Effectors of fatty acid synthesis in hepatoma tissue culture cells. PMID- 6284037 TI - Intracellular catalase function: analysis of the catalatic activity by product formation in isolated liver cells. PMID- 6284038 TI - Paramagnetism in melanins: pH dependence. PMID- 6284039 TI - Analysis of cyclic nucleotide phosphodiesterase(s) by radioimmunoassay. PMID- 6284040 TI - Regulation of low-density lipoprotein receptors in cultured bovine adrenocortical cells. PMID- 6284041 TI - Effect of CuSO4 and Cu(II)(Gly)2 on some indirect assays for superoxide dismutase activity. PMID- 6284042 TI - How does limited trypsinization influence the surface structure and binding of calcium to lipoprotein (a): a spin-labeling study. PMID- 6284043 TI - Partial purification from hepatoma cells of an intracellular substance which mediates the effects of insulin on pyruvate dehydrogenase and low Km cyclic AMP phosphodiesterase. PMID- 6284045 TI - The role of superoxide anion in peroxidase-catalyzed chemiluminescence of luminol. PMID- 6284044 TI - Isolation of Paracoccus denitrificans cytochrome cd1: comparative kinetics with other nitrite reductases. PMID- 6284046 TI - Catalase synthesis in Escherichia coli is not controlled by catabolite repression. PMID- 6284047 TI - A method for the detection of superoxide in biological systems. PMID- 6284049 TI - Interaction of concanavalin A with lactogenic receptors in isolated membranes. PMID- 6284048 TI - Studies on the in vitro phosphorylation of 6-phosphofructo-1-kinase from rat liver. PMID- 6284050 TI - Relationship between H+, anion, and monovalent cation movements and Ca2+ transport in sarcoplasmic reticulum: further proof of a cation exchange mechanism for the Ca2+-Mg2+-ATPase pump. PMID- 6284051 TI - Acetylcholine receptor-mediated sodium ion efflux after rapid hypoosmotic loading of radiotracer. PMID- 6284052 TI - Equilibrium constants under physiological conditions for the reactions of L phosphoserine phosphatase and pyrophosphate: L-serine phosphotransferase. PMID- 6284053 TI - Amphiphile-mediated activation of soluble adenylate cyclase of Bordetella pertussis. PMID- 6284054 TI - Cytochrome levels and activity in stored human platelets. PMID- 6284055 TI - Iron-ethylenediaminetetraacetic acid (EDTA)-catalyzed superoxide dismutation revisited: an explanation of why the dismutase activity of Fe-EDTA cannot be detected in the cytochrome c/Xanthine oxidase assay system. PMID- 6284056 TI - The identification of a serum viability factor for SV3T3 cells as biotin and its possible relationship to the maintenance of Krebs cycle activity. PMID- 6284057 TI - Investigations of the state of the manganese in Lactobacillus plantarum. PMID- 6284058 TI - Adrenocortical cyclic GMP-dependent protein kinase: purification, characterization, and modification of its activity by calmodulin, and its relationship with steroidogenesis. PMID- 6284060 TI - Improvement of psoriasis of the scalp with ketoconazole. PMID- 6284059 TI - Purification and properties of poly(ADP-ribose) synthetase from mouse testicle. PMID- 6284061 TI - Different papillomaviruses as the causes of oral warts. AB - We have observed four patients with oral papillomas. Two children had oral mucosal lesions characteristic of focal epithelial hyperplasia, a young man had common, wart-like lesions on his hard palate, and a male immunosuppressed renal allograft recipient had condyloma-like lesions on his gingivae. Papillomavirus like particles were seen by electron microscopy in lesions from both patients with focal epithelial hyperplasia. No structural antigens for human papillomavirus (HPV) 1, 2, 3, or 5 were found by immunofluorescent microscopy, but further evidence of the presence of a papillomavirus was found by immunoperoxidase microscopy using a cross-reacting sodium lauryl sulfate disrupted bovine papillomavirus 1 anti-rabbit serum sample. The distinct histologic pattern seen in focal epithelial hyperplasia suggests that a yet undescribed HPV type might be associated with this disease. Histologic, ultrastructural, and immunofluorescent microscopy and restriction endonuclease analysis all gave evidence of HPV 2 in the palatal lesions in patient 3. Evidence of papillomavirus antigen was found by immunoperoxidase microscopy in the oral condylomas from our immunosuppressed patient. PMID- 6284063 TI - Ulcerative contact dermatitis caused by sodium silicate. Coexistence of primary irritant contact dermatitis and contact urticaria. AB - A 57-year-old man had had recurrent ulcerative lesions on his left hand for two years. The ulcers were associated with chronic eczematous changes resulting from primary irritant contact dermatitis to sodium silicate, as indicated by positive patch tests. The patient also had another type of cutaneous reaction to sodium silicate, ie, contact urticaria. An immediate wheal and flare reaction was found 15 minutes after application of sodium silicate to a scratch test site. This response was not seen in healthy control subjects. To our knowledge, the coexistence of primary irritant contact dermatitis and contact urticaria, both induced by sodium silicate, has not previously been described. PMID- 6284062 TI - Tissue angiotensin-converting enzyme and lysosomal enzyme levels in skin diseases. AB - The activities of tissue angiotensin-converting enzyme (ACE) were investigated in several skin diseases. Biopsy specimens were taken from 49 patients with granulomatous lesions, including sarcoidosis, 16 patients with nongranulomatous lesions, and 13 normal individuals. The ACE level was measured fluorometrically and the lysosomal enzyme levels were measured by established techniques. It was found that ACE activity increased in all granulomatous skin lesions, but not in other inflammatory lesions. The enzyme activity showed an increase that paralleled the time course of the disease. In contrast, lysosomal enzymes increased in various types of skin lesions, suggesting that they reflect nonspecific inflammation. These findings indicate that the tissue ACE level may be a marker for granulomatous inflammation, but one that is not specific for sarcoidosis. PMID- 6284064 TI - Pathology quiz case 2. Eccrine spiradenoma. PMID- 6284065 TI - [Malignant giant cell fibrous histiocytoma of the soft parts]. PMID- 6284066 TI - [Clinical, epidemiological, morphological and etiopathogenetic aspects of benign tumors of hepatocytic derivation, with description of a case of focal nodular hyperplasia]. PMID- 6284067 TI - Movement and identification of a creosote-derived PAH complex below a river pollution point source. AB - High pressure liquid chromatography was used to determine the identity of a black, oily leachate seeping into a spring-fed river. The leachate was identified as creosote by comparing a sample of the material to a sample of commercial creosote. As creosote contains a complex mixture of polynuclear aromatic hydrocarbons (PAH), the direct introduction of creosote into a stream system provided an opportunity to study the kinetics of these hydrocarbons as sediment contaminants below the creosote source. Concentrations of PAH, calculated on the basis of the amounts of organic matter contained in the sediments, declined rapidly below the point source in a form readily approximated by double exponential equations. An important aspect of the pattern of decline encountered was that concentrations tended to become asymptotic. As a result, the river's sediments became contaminated over a considerable distance. PMID- 6284068 TI - Effects of benzo(a)pyrene on early development of flatfish. AB - The ontogenetic effects of the environmental carcinogen benzo(a)pyrene (BAP) on three species of larval flatfish were investigated using concentrations (from 0.10 to 4.2 ppb) which were comparable to levels found in polluted harbors. BAP treated sand sole (Psettichthys melanostichus) eggs displayed a significant decline in hatching success and a significantly higher incidence of developmental anomalies than did control eggs. Flathead sole (Hippoglossoides elassodon) eggs exposed to a single dose of a water-soluble BAP-bovine serum albumin complex demonstrated evidence of toxic injury with pycnotic nuclei present in the integument and, more commonly, in ocular and neural tissues. An increased incidence of morphological anomalies in English sole (Parophyrs vetulus) eggs and larvae exposed to BAP was not detected. PMID- 6284069 TI - Serial PBB levels, PCB levels, and clinical chemistries in Michigan's PBB cohort. AB - Half of a cohort of 3683 Michigan residents exposed to polybrominated biphenyls (PBBs) in 1973 and 1974 had 2 serum PBB determinations at a 1- or 2-yr interval. The median decrease in serum PBB levels during both 1- and 2-yr intervals was 1 microgram/L. The geometric mean serum polychlorinated biphenyl (PCB) level (6.3 microgram/L) exceeded that of PBB (4.1 micrograms/L), although the range of PCB levels (smaller than 1-57 microgram/L) was narrower than that of PBB levels (smaller than 1-3150 micrograms/L). Mean PCB and PBB levels were higher for males, and mean PCB levels increased with age. In a subgroup with higher-than average PBB levels, serial clinical chemistry tests during 4 different years showed no consistent significant correlation with serum PBB levels. Tests with greater sensitivity and specificity for hepatic microsomal enzyme induction and thyroid status are needed in future evaluations of the most highly exposed subgroup of the cohort. PMID- 6284070 TI - Environmental fiber-induced pleuro-pulmonary diseases in an Anatolian village: an epidemiologic study. AB - This study was designed to determine the prevalence of pleural mesothelioma and other malignancies in the Anatolian village of Tuzkoy, where neither asbestos nor any environmental carcinogen has been detected. Another village (Kizilkoy) located 12 km from Tuzkoy was selected as a control. Three hundred twelve subjects from Tuzkoy who were at least 25 yr of age and 95 subjects from Kizilkoy were studied. Analysis of X-rays of the Tuzkoy group revealed that subjects had calcified pleural plaques (17%), pleural thickening (10.5%), obscured costophrenic angles (15%), and diffuse interstitial pulmonary fibrosis (12.1%). Sixty-seven deaths were records in Tuzkoy during the previous 3 yr, 41 of which resulted from malignant diseases. There were no X-ray abnormalities or deaths resulting from malignancies in the control group. Because of the high incidence of mesothelioma and lung cancer which usually results from asbestos exposure, the presence of asbestos in Tuzkoy was investigated, but none was detected in spite to Tuzkoy's volcanic location. Nevertheless, zeolite, an asbestiform mineral, was detected in the stones of buildings and in the village soil, as well as in the lung and pleura of the patients during biopsy. Thus, this mineral was considered to be responsible for the fiber-induced pleuro-pulmonary diseases in Tuzkoy. No zeolite was found in the soil and stones of the control village. PMID- 6284071 TI - An ultrastructural analysis of breast carcinoma presenting as isolated axillary adenopathy. AB - Metastatic adenocarcinoma in the axillary lymph nodes of a female patient often originates from a primary tumor in the ipsilateral breast. Mastectomy may be recommended if adenocarcinoma is found in the axillary nodes even when the primary tumor is not clinically detectable. In these circumstances, the recommendation for mastectomy should be based on the firm histologic diagnosis of adenocarcinoma. In the present report, five female patients are discussed who presented with axillary lymphadenopathy without clinically evident breast masses or mammographic evidence of malignancy. Axillary lymph node biopsies, performed in each patient, were inconclusive after conventional light microscopic examination. Electron microscopy established the diagnosis of adenocarcinoma. These findings were complemented by sex steroid analyses of the tumors where possible. Each patient underwent ipsilateral mastectomy, and in each specimen an occult breast carcinoma was found. The necessity of making a precise tissue diagnosis in all cases of metastatic cancer from an unknown primary is stressed, and special techniques to accomplish this must be considered preoperatively. This is particularly important in the female patient with metastatic breast carcinoma in an isolated axillary lymph node, since ipsilateral mastectomy may be curative. PMID- 6284072 TI - Splenic capping: an experimental study of a new technique for splenorrhaphy using woven polyglycolic acid mesh. AB - The use of polyglycolic acid (PGA) stretchable mesh applied to the experimentally injured canine spleen can achieve satisfactory immediate hemostasis by tamponade and simplifies the use of sutures to control remaining areas of hemorrhage. PGA mesh with 1/4" and 1/8" openings was utilized for splenorrhaphy in 12 adult mongrel dogs subjected to sharp splenic trauma. By gross and histologic examination, the PGA mesh material appears to undergo progressive absorption to complete absorption by 85 days. For the 12 animals and 30 operative procedures the only complications of the use of the mesh were the occurrence of an intrasplenic seroma in a single animal at 6 weeks after operation and three wound infections. With this material, rapid, simple canine splenic injury repair can be achieved. PGA mesh further assists in the healing process, and in maintaining maximum splenic architecture and function. The material used in this study was manufactured and supplied by Davis & Geck, American Cyanamid, Danbury, Connecticut. PMID- 6284075 TI - The epidemiology of O'nyong-nyong in the Kano Plain, Kenya. PMID- 6284073 TI - Infusion of branched-chain enriched amino acid solution in patients with hepatic encephalopathy. AB - Hospitalized patients with hepatic insufficiency often suffer from severe catabolic states and are in urgent need of nutritional support during their acute illness. Protein intolerence, however, remains a significant problem with respect to the provision of adequate nutrition, either enterally or parenterally. The following report is an anecdotal series of 63 consecutive patients in a large urban hospital treated prospectively with nutritional support using a prototype high branched-chain amino acid solution (FO80) given by technique of total parenteral nutrition by the subclavian or internal jugular route with hypertonic dextrose. Sixty-three patients, of which 42 had chronic liver disease (cirrhosis) with acute decompensation and 17 with acute hepatic injury as well as four with hepatorenal syndrome, are the subject of this report. All required intravenous nutritional support and were either intolerant to commercially available parenteral nutrition solutions or were in hepatic encephalopathy at the time they were initially seen. The cirrhotic patients had been hospitalized for a mean of 14.5 +/- 1.9 days before therapy, had a mean bilirubin of 13 mg/100 ml, and had been in coma for 4.8 +/- 0.7 days despite standard therapy. Patients with acute hepatitis had been in the hospital for 16.2 +/- 4.1 days before therapy, had a mean bilirubin of 25 mg/100 ml, and had been in coma 5.2 +/- 1.6 days before therapy. Routine tests of liver function, blood chemistries, amino acids, EEGs, and complex neurological testing including Reitan trailmaking tests were used in the evaluation of these patients. Up to 120 grams of synthetic amino acid solution with hypertonic dextrose was tolerated in these patients with improvement noted in encephalopathy of at least one grade in 87% of the patients with cirrhosis and 75% of the patients with hepatitis. Nitrogen balance was achieved when 75 to 80 grams of synthetic amino acids were administered. Survival was 45% in the cirrhotic group and 47% in the acute hepatitis group. Encephalopathy appeared to correlate with individual amino acids differentially in the various groups and with the ratio between the aromatic and the branched chain amino acids. Ammonia did not correlate with either the degree of encephalopathy or improvement therefrom. In 24 Patients therapy for hepatic encephalopathy was limited to infusion of the branched-chain enriched amino acid solution only, with wake-up in 66% of this group. The results strongly suggest that in protein intolerant patients requiring nutritional support, infusion with branchedchain enriched amino acid solutions is well tolerated with either no worsening of or improvement in hepatic encephalopathy coincident with the achievement of nitrogen equilibrium and adequate nutritional support. PMID- 6284074 TI - Sero-epidemiology of amoebiasis in Peninsular Malaysia. PMID- 6284077 TI - Radiation-induced soft-tissue fibrosarcoma: surgical therapy and salvage. PMID- 6284078 TI - Opiate receptor involvement in the quasi-morphine withdrawal syndrome induced by acute injection of R 18 503. PMID- 6284079 TI - Alpha 1- and alpha 2-adrenoceptor stimulation and Ca fluxes in isolated rat aorta. PMID- 6284076 TI - Bronchogenic carcinoma: a study of patients treated at Capital Hospital, Peking, China. AB - From 1961 to 1972, 123 patients with lung cancer underwent operations at Capital Hospital, Peking, China. Ninety-six patients had resectable lesions and 27 did not, a resectability rate of 78%. Four patients (4.2%) died immediately postoperatively. Complications occurred in 8 (8.3%) patients. Five-year survival in this group of 92 survivors was 26 (28.3%). Ten-year survival among 49 patients was 10 (20.4%). Among the 92 five-year survivors, 41.2% had squamous cell carcinoma, 25% had adenocarcinoma, and 16% had undifferentiated carcinoma. Patients with squamous cell carcinoma had a much longer survival than the others. Six factors appear to influence survival after resection: cell type, presence of lymph node metastases, presence of tumor emboli in blood vessels, sex, age, and location. Men about 50 years of age, with a peripherally located squamous cell tumor and with no tumor emboli or lymph node metastases, have a good chance of surviving a pulmonary resection for ten years. PMID- 6284080 TI - Clofibrate, calcium and cardiac muscle. AB - The anti-hyperlipidemic drug clofibrate produces negative inotropic effects and arrythmias in isolated perfused rabbit heart Langendorff preparations. In electrically stimulated rat left atria, clofibrate produces negative inotropic effects, the speed of onset and extent of which are decreased by raising the Ca concentration of the bathing medium. Sensitivity of isolated rat atria to clofibrate is not increased when the tissues are stimulated under slow Ca channel conditions, in which the tissues are activated by either isoproterenol or dibutyryl cyclic AMP, although sensitivity to clofibrate is decreased when atria are exposed to increasing concentrations of norepinephrine. Increasing the stimulation frequency of isolated guinea-pig atria to produce a positive treppe also decreases the inhibitory effect of clofibrate, while in rat atria the typical negative treppe is altered towards a positive treppe in presence of clofibrate. The effects of paired electrical stimulation are not diminished by the drug, suggesting that Ca release from the sarcoplasmic reticulum is not affected by clofibrate, although the drug inhibits the rate of Ca uptake by isolated cardiac sarcoplasmic reticulum and mitochondria. These results suggest that clofibrate has multiple effects on Ca functions in cardiac muscle. PMID- 6284081 TI - Low-dose captopril: its use in mild to moderate hypertension unresponsive to diuretic treatment. AB - The effect of low doses (25 mg three times a day) of captopril was evaluated in 16 patients with mild to moderate essential hypertension, previously uncontrolled by hydrochlorothiazide. After a no-treatment period, mean eight-hour seated diastolic blood pressure (SDBP, mm Hg) was 103 +/- 5 on placebo, 95 +/- 8 after a single dose of captopril, 96 +/- 4 after two weeks of captopril alone, and 90 +/- 6 after its combination with hydrochlorothiazide. Though nine patients had at least a 10% fall in SDBP after the initial dose of captopril, only three had a comparable fall after two weeks; after captopril and hydrochlorothiazide, however, 12 patients had such a response. Captopril decreased mean angiotensin converting enzyme activity and plasma aldosterone, though to a lesser extent with continued therapy. Because its side effects appear dose related, low doses of captopril combined with a diuretic are effective and may be better tolerated. PMID- 6284082 TI - Seroconversion to Entamoeba histolytica among short-term travelers to Mexico. AB - Paired serum samples were evaluated for the presence of antibodies to Entamoeba histolytica in 326 healthy college students from the United States who lived in Guadalajara, Mexico, for an average of four weeks. One hundred eighty of these students had an enteric illness develop, but no stool test results were positive for ameba. An indirect hemagglutination assay was the serologic method used, and no seroconversion was demonstrates. This finding confirms the belief that amebiasis is a rare cause of diarrhea among short-term travelers to Mexico. PMID- 6284083 TI - Enterococcal endocarditis: a rare cause of focal cardiac uptake in infarct-avid myocardial scintigraphy. PMID- 6284084 TI - Fibrosing uremic pleuritis complicated by thoracic empyema. PMID- 6284085 TI - Immunoperoxidase localization of lactalbumin in malignant breast neoplasms. AB - By an immunoperoxidase method, lactalbumin was demonstrated in the cytoplasm of epithelial cells of normal breast, primary and metastatic breast carcinomas, and benign breast disease. Lactalbumin was present in 39 of 64 breast carcinomas of different types, including 77% (31) of infiltrating ductal carcinomas and 42% (seven) of infiltrating lobular carcinomas. It also was present in 55% (nine) of metastatic breast carcinomas and in 100% (five) of fibroadenomas. Lactalbumin was not observed in normal tissues and malignant neoplasms of other tissues, indicating that lactalbumin can serve as a specific tissue marker for malignant neoplasms of the breast and their metastases. PMID- 6284086 TI - Massive articular chondrocalcinosis. Its occurrence with calcium pyrophosphate crystal deposits in nucleus pulposus. AB - We studied a case of long-standing chondrocalcinosis that after 40 years affected almost all fibrocartilages, hyaline cartilages, and articular cavities within the subject. Massive deposits of dihydrocalcium pyrophosphate (CPP) were also found in nonfissured nucleus pulposus of the vertebral disks. To our knowledge, this is the first report of CPP deposits in these disks. PMID- 6284087 TI - Rectal carcinoma and coexistent primary hepatocellular carcinoma. PMID- 6284088 TI - Peripheral neuropathy after multiple tetanus toxoid injections. AB - This case documents the 14th reported patient with peripheral neuropathy following tetanus toxoid injection. A 33-year-old man developed profound mixed sensorimotor peripheral neuropathy after receiving 2 tetanus toxoid injections over a 5-month period. Periodic serial electromyographic and nerve conduction studies performed over 2 years suggested both segmental demyelination and axonal neuropathy. The patient experienced partial recovery. Analysis of this case and 13 others reported in literature indicates that in almost all cases (85%), patients had received multiple tetanus toxoid injections. Also, it appears that a prolonged interval of 14 or more days between the tetanus toxoid injection and the onset of neurological symptoms is associated with a poorer prognosis for complete recovery. PMID- 6284089 TI - Mastectomy as an adjuvant to chemotherapy for locally advanced or metastatic breast cancer. PMID- 6284090 TI - Neurochemical changes in rats coexposed to lead and copper. AB - Rats were coexposed to lead (Pb) and Copper (Cu) through drinking water and intraperitoneally, respectively, for a period of 21 days. Neurochemical studies in these rats showed significant reduction in the activity of adenosine triphosphatase, cytochrome-c-oxidase, diaphorase and in the levels of biogenic amines in the rats simultaneously exposed to the two metals compared to either of the metal alone. These neurotoxic effects were not related to the contents of either of the metals in the brain since their accumulation after combined exposure was much less than observed after individual exposure to Pb or Cu. PMID- 6284092 TI - Effect of ribosome-inactivating proteins on virus-infected cells. Inhibition of virus multiplication and of protein synthesis. AB - HEp-2 cells were infected with herpes simplex virus-1 (HSV-1) or with polio-virus I in the presence of plant proteins which inactivate ribosomes in cell-free systems, while exerting scarce effect on whole cells. Ribosome-inactivating proteins used were gelonin, from seeds of Gelonium multiflorum, an inhibitor from the seeds of Momordica charantia, dianthin 32, from the leaves of Dianthus caryophyllus (carnation), and PAP-S, from the seeds of Phytolacca americana (pokeweed). All proteins tested had the following effects: 1. They reduced viral yield; 2. They decreased HSV-1 plaque-forming efficiency; 3. They inhibited protein synthesis more in infected than in uninfected cells. These results strongly suggest that ribosome-inactivating proteins impair viral replication by inhibiting protein synthesis in virus-infected cells, in which presumably they enter more easily than in uninfected cells. PMID- 6284093 TI - Variations in the buoyant density of foot-and-mouth disease virus strains. AB - Some of the factors which effect the buoyant density in caesium chloride of foot and-mouth disease virus were investigated. Under standard conditions, the buoyant densities of a range of vaccine virus strains, representing all seven serotypes, were found to be unrelated to their potential of being formulated into effective vaccines. In some virus strains, a small amount of naturally occurring high and/or low density components were observed and these components were examined in more detail in one virus strain. PMID- 6284091 TI - Neutralising antibody in mice with primary and recurrent herpes simplex virus infection. AB - Neutralising antibody was studied in mice infected in the ear with herpes simplex virus type 1. Antibody was detected six days after infection and after one month it had reached titres which subsequently varied little. Geometric group mean titres were doubled by reinoculation of virus whereas recurrent disease induced by trauma did not change them (although titres did change in some individual animals). Mean titres in mice which had had unequivocal signs of primary disease were twice as high as in those which had not, and the frequency of recurrent disease in response to trauma in the former animals was greater than in the latter. A reduction in the frequency of induced recurrent disease was seen if the application of trauma was delayed for some time after the establishment of latency. Hyperimmunisation increased mean titres tenfold. If given soon after primary infection it reduced the frequency of induced recurrent disease but if its administration was delayed then the frequency was increased. PMID- 6284094 TI - Actin distribution and synthesis in human fibroblasts infected by cytomegalovirus. AB - "Early functions" of human cytomegalovirus were found to induce a loss of microfilaments as revealed by indirect immunofluorescence and by the use of fluorescent phalloidin. Actin synthesis in infected cultures, on the other hand, appeared to be largely unchanged as estimated from the specific radioactivity of cytoplasmic actin. PMID- 6284095 TI - Somatosensory evoked response: application in neurology. AB - One technique used for short-latency somatosensory evoked response (SER) is described. SER following nerve stimulation is a unique non-invasive, clinical test used to evaluate the somatosensory pathways. It tests the physiological function of the median nerve, the brachial plexus, the C6-7 cervical roots, cervical spinal cord, the cuneate nuclei, the medial lemniscus, the thalamus, and the contralateral sensory cortex. It has been shown to be a reliable and useful clinical test particularly in multiple sclerosis and comatose patients. The promising technique of SER following peroneal nerve stimulation is mentioned. PMID- 6284096 TI - Neural pathway involved in the excitation of motoneurones to jaw-opening muscles by stimulation of the orbital cerebral cortex in the cat. AB - The pathway and synaptic basis for the efferent discharge in the digastric nerve evoked by stimulation of the orbital cortex were studied in cats anesthetized with alpha-chloralose. Transection of the brain stem at the boundary between the pons and the medulla oblongata abolished discharge. Intracellular recording from anterior digastric motoneurones revealed that the orbital cortical stimulation evoked excitatory post-synaptic potentials with a latency that was longer by one synaptic delay than that of spike potentials evoked in medial bulbar reticular neurones making a direct excitatory projection to anterior digastric motoneurones. It was concluded that the excitatory effects was mediated by the medial bulbar reticular formation. PMID- 6284097 TI - Proton beam irradiation of uveal melanomas. Results of 5 1/2-year study. AB - Proton beam irradiation was used in the treatment of 76 uveal melanomas from July 1975 to December 1980. Five (7%) were small, 32 (42%) were medium, and 39 (51%) were large melanomas. The follow-up period ranged from two months to 5 1/2 years; 19 patients were followed up for more than two years and 39 were observed for more than a year. Tumor regression has been achieved in all eyes with more than 12 months of follow-up except one, which was enucleated because of secondary complications. Three patients in whom metastatic disease developed died. Our data indicate that proton irradiation can be used for the treatment of relatively large lesions that previously were considered untreatable and reduces the high ocular morbidity experienced with other methods in the treatment of medium and small melanomas. PMID- 6284098 TI - Glomus Tumors. Diagnosis, classification, and management of large lesions. AB - Glomus tumors of the temporal region and skull base can range in size from small microscopic lesions confined to the promontory to large destructive and neurologically aggressive agents of substantive incapacitation. Contemporary diagnostic and surgical technology has made definitive management of these historically dreaded management of these historically dreaded lesions confidently practical. Prompted by these advances, a new classification of glomus tumors is disclosed. The specific problem of the large chemodectoma at the skull base is addressed and a series of such cases reviewed. Surgical therapy is viewed as definitive, and the technique employed by the members of The Otology Group, PC, is described. PMID- 6284099 TI - Prostacyclin and arterial wall biology. PMID- 6284100 TI - Dietary lipids, sugar, fiber and mortality from coronary heart disease. Bivariate analysis of international data. AB - Univariate analyses with data for the years 1954-1969 from 20 economically advanced countries showed that a combined dietary lipid score based on per capita consumption of saturated fat, cholesterol, and polyunsaturated fat (equation of Keys, et al.) and the consumption of refined and processed sugars were both highly correlated with age-standardized coronary heart disease (CHD) mortality rates for the years 1969-1973 for both men and women aged 35-74 years (r values of 0.54 to 0.72). Fiber intake, estimated as the sum of calories available from vegetables, fruits, grains, and legumes, yielded a significant inverse correlation with CHD mortality rates (r = -0.49 to -0.68). These three dietary variables are highly intercorrelated. With bivariate analyses (2 X 2 cross classification and analysis of variance), lipid score, but neither that of sucrose or fiber, was consistently and significantly related to CHD mortality. However, with both higher and lower lipid scores, CHD mortality rates tended to be slightly (nonsignificantly) higher with higher sugar values and with lower fiber values. PMID- 6284101 TI - Results of a system of registration and follow-up of patients with hydatidiform mole. PMID- 6284102 TI - Thyroid hormone binding to putative nuclear receptors in human mononuclear blood cells. AB - Nuclear protein extracts from mononuclear cells in normal subjects, subjects with the clinical syndrome of thyroid hormone resistance and cultured B lymphocytes bound thyroxine (T4) Ka; 1 . 3-3 . 5 X 10(9) m-1) and triiodothyronine (T3) (Ka; 1 . 2-3 . 9 X 10(9) m-1) with similar affinities. When an extra wash step was introduced, the maximum specific binding of T4 and T3 was reduced by 75% and binding was abolished by three washes. These data suggest binding is to a serum protein contaminant present in the mononuclear cell preparation. PMID- 6284103 TI - Models for the metabolic production of oxalate from xylitol in humans: a role for fructokinase and aldolase. AB - It has been proposed previously that oxalate precursors may be formed in the transketolase reaction during the metabolism of xylitol. It is shown in this paper that fructokinase and aldolase, purified from human liver, provide an alternative model in that, in coupled sequence, they produce glycolaldehyde, an oxalate precursor, from D-xylulose via D-xylulose 1-phosphate; D-fructose does not give rise to glycolaldehyde. It is concluded that metabolic pathways based on a combination of the transketolase, fructokinase and aldolase reactions can account for the production of glucose, lactate, tetronates (C-threonic and D erythronic acids) and oxalate (precursors) during the metabolism of xylitol administered parenterally. PMID- 6284104 TI - A preliminary study on the pathogenicity of axenically grown Entamoeba histolytica for the golden hamster. PMID- 6284105 TI - Hypophosphataemic vitamin D-resistant rickets--a cause of spinal stenosis in adults. PMID- 6284106 TI - Resistance to Marek's disease at hatching in chickens vaccinated as embryos with the turkey herpesvirus. AB - Chickens vaccinated with herpesvirus of turkey (HVT) as 18-day embryos or at hatching were challenged as neonates with pathogenic Marek's disease (MD) virus (MDV). Embryonally vaccinated chickens had much greater resistant to challenge than chickens vaccinated post-hatch. Embryos became readily infected with HVT regardless of whether the vaccine was deposited into the body of the embryo or extraembryonally, such as in the amniotic sac. Embryonally vaccinated chickens were viremic with HVT at hatching and remained persistently viremic through the duration of the experiment. The titer of recoverable virus was higher in the embryonally vaccinated chickens than in the chickens vaccinated post-hatch. Embryonal vaccination did not affect hatchability. Vaccination at any stage of embryonation tested protected better against neonatal challenge than did vaccination at hatching. Protection against an early challenge was greatest when the embryos were 17 or 18 days old at the time of vaccination. Lower protection in chickens vaccinated as 11-day embryos was not due to humoral immunologic tolerance. Chickens vaccinated at the 11th day of embryonation were poorly protected against MDV challenge at three or eight days of age but were well protected if the challenge was delayed until the 14th day of age. PMID- 6284107 TI - In vitro studies of hemorrhagic enteritis virus with immunofluorescent antibody technique. AB - Dissociated spleen lymphocytes from hemorrhagic enteritis virus (HEV)-free chickens, pheasants, and turkeys were infected with HEV in suspension cultures. Viral antigens were detected in cells of all three donor species by immunofluorescence. The age of chicken from which cells were obtained and the temperature of incubation influenced the development of viral antigens in cultured cells, which could be observed as early as 24 hr postinfection. This is the first time that HEV has been shown to infect cells in vitro. PMID- 6284108 TI - Immunofluorescence studies on the early pathogenesis of hemorrhagic enteritis virus infection in turkeys and chickens. AB - Chickens and turkeys not previously exposed to hemorrhagic enteritis virus (HEV) were inoculated orally with the virus. Birds were necropsied each day from the first to the eighth day following inoculation, and specimens from various tissues were collected for examination. Clinical illness was noted only in turkeys, although intestinal hemorrhages and swollen, necrotic spleens were seen in both species. The distribution and localization of viral antigens in various tissues and peripheral blood lymphocytes were studied by the immunofluorescent technique. The patterns of HEV infection in chickens and turkeys were similar, except there was a high level and persistence of viral antigen in the thymus of turkeys but not in chickens. PMID- 6284109 TI - Effect of Mycoplasma gallisepticum on genetically defined chickens. AB - Three parameters were used to measure differences between Arkansas progressor and regressor lines of chickens in response to Mycoplasma gallisepticum inoculations. The responses of regressor and progressor lines to M. gallisepticum did not differ as judged by antibody response, severity of airsacculitis, and resistance of tracheal ring tissue cultures from 20-day-old progressor and regressor chick embryos to M. gallisepticum. PMID- 6284110 TI - Enzyme-linked immunosorbent assay for the detection of antibodies to infectious bronchitis virus. AB - An enzyme-linked immunosorbent assay (ELISA) for antibodies to avian infectious bronchitis (IB) virus is described. The immune response of chickens following vaccination with IB virus was monitored using this test, and the titers were compared with those obtained by serum neutralization. The ELISA appears to be suitable for IB serology. PMID- 6284111 TI - Pathogenicity of infectious laryngotracheitis virus as measured by chicken embryo inoculation. AB - A comparison was made between pathogenicities for chicken embryos of unattenuated and attenuated strains or isolates of infectious laryngotracheitis (ILT) virus. All 11-day-old chicken embryos inoculated with 10(3.0) or 10(4.0) TCID50 of unattenuated strain NS175 via allantoic cavity died within 6 days. On the contrary, no chicken embryos of the same age died when inoculated with the same amount of cell-culture-attenuated isolate C7 in a like manner. The mortality index for chicken embryos (MICE) was obtained by dividing the cumulative number of embryos dying within 7 days by the cumulative number of embryos surviving 7 days. The reliability of the MICE test was confirmed by duplicate and triplicate experiments with strain NS175 and isolate C7. MICE obtained in the experiments with 9 different strains or isolates of ILT virus ranged from 0 to 1, and the values were well correlated with the pathogenicities for chickens. The results from the present work suggest that strains or isolates with MICE less than 0.16 would have low or no pathogenicity for chickens, and those with MICE more than 0.27 would be highly pathogenic. Further studies are needed using additional isolates of ILT virus with varied pathogenicities. PMID- 6284112 TI - A simple technique for preparation of chicken-embryo-skin cell cultures. AB - A simple, rapid technique was developed for preparing chicken-embryo-skin cell cultures utilizing trypsinization of the skin of intact 12-day-old chicken embryos. When cell cultures were inoculated with fowl pox virus, those that consisted of at least 80% epithelial cells yielded a higher virus titer than fibroblast cell cultures. PMID- 6284113 TI - Superinfection in ducks persistently infected with duck plague virus. AB - Superinfections with homologous or heterologous strains of duck plague virus resulted in the deaths of birds persistently infected with duck plague virus. Not all birds that were superinfected died. Protection against mortality depended on the route of exposure, strain of the initial duck plague virus, and strain of the superinfecting virus. PMID- 6284114 TI - The immune response of the chick following viral vaccinations and immunization with sheep red blood cells. AB - The effects of viral vaccinations and immunization with sheep red blood cells (SRBC) on the humoral response of pullets were investigated. Pullets were vaccinated with Marek's disease virus, Newcastle disease virus (NDV), infectious bronchitis virus (IBV), and infectious bursal disease virus at appropriate ages used in commercial practice. At seven weeks, the pullets were intramuscularly immunized with SRBC. NDV and IBV antibodies were detected by hemagglutination inhibition tests. Hemagglutination (HA) titers were established against SRBC. IBV antibody titers were not affected by vaccination or by immunization with SRBC. NDV antibody titers were significantly increased by vaccination and by immunization with SRBC. The SRBC agglutinin response was also positively affected by vaccination. The HA titer increase consisted of a rise in 2-mercaptoethanol (2 ME)-sensitive antibodies and a fall in 2-ME-resistant antibodies. PMID- 6284115 TI - Monoamine oxidase (MAO) activity as a determinant in human neurophysiology. AB - Several lines of evidence indicate that monoamine oxidase (MAO) activity can regulate levels of biogenic amines and neuronal activity in the nervous system. The two types of MAO activity, A and B, appear to have different domains of activity in the body. Brain tissue has both types of activity, although adrenergic neurons are thought to contain exclusively MAO-A. MAO activity can also be measured in peripheral tissues: MAO-A in cultured skin fibroblasts and placenta, and MAO-B in platelets and lymphocytes. These two types of activity are mediated by different enzyme molecules and are regulated independently by endogenous and exogenous factors including genetic determinants, hormones, and aging. In humans, inhibition of MAO-A activity leads to mood elevation in depressed patients; in contrast, low MAO-B activity in platelets has been associated with an increased susceptibility to psychopathology. In order to assess further the role of MAO activity in human mood and behavior, it will be important to measure both forms of the enzyme independently and to establish correlations between levels of activity and discrete phenotypic traits. PMID- 6284118 TI - [Idiopathic lumbar spinal stenosis]. PMID- 6284116 TI - Opioid peptides as neuroregulators: potential areas for the study of genetic behavioral mechanisms. AB - The opioid peptides have been related to behavior in both animal and human studies. Further investigation can be anticipated which could lead to the elucidation of genetic controls over enzymes which process these peptides and the receptors upon which the peptides act. The enzymes, both synthetic and degradative, can lead to the formation of different forms of the opiate peptides. Differential control of these enzymes or of the multiple forms of opiate receptors could lead to discrete changes in opiate status and subsequent behavioral changes. Conversely, genetically regulated behavioral modification could also lead secondarily to opiate changes. PMID- 6284119 TI - [Narrow lumbar spinal canal]. PMID- 6284120 TI - [Cases of Paresis and Paralysis in a German Thoroughbred Stud (author's transl)]. PMID- 6284117 TI - Calcium movements in relation to heart function. PMID- 6284122 TI - The binding of platinum complexes to tuna cytochrome c. AB - The binding of [PtCl4]2- and cis-[PtCl2(NH3)2] to methionine-65 of tuna cytochrome c was investigated by 1H n.m.r. The modification at methionine-65 is shown to cause an extremely small structural perturbation to the protein at the site of modification. PMID- 6284123 TI - Hydrolysis of monomolecular layers of synthetic sphingomyelins by sphingomyelinase of Staphylococcus aureus. AB - The enzymic hydrolysis of three synthetic sphingomyelins, spread as monomolecular films at the air/water interface by purified Staphylococcus aureus sphingomyelinase was studied. Each of the three sphinomyelins (DL-erythro-N palmitoyl-, -N-stearoyl- and -N-lignoceryl-sphingosylphosphocholine) has an optimal activity-dependent surface pressure or concentration curve. The optimal surface pressure as well as the optimal surface density for hydrolysis was different for each of the three substrates. This optimum coincides with the liquid-condensed/liquid-expanded phase transition for each of the sphingomyelins. At initial surface pressures (pi 0) below the optimum, reaction rates are controlled mainly by surface density of the substrate; above the optimal pi 0, reaction rates decrease with increasing surface pressure. The difference between the three synthetic sphingomyelins are explained by the variation in the degree of asymmetry between their two paraffinic chains. PMID- 6284124 TI - Studies of the conformation of bilirubin and its dimethyl ester in dimethyl sulphoxide solutions by nuclear magnetic resonance. AB - The conformation of bilirubin and its dimethyl ester in dimethyl sulphoxide (DMSO) was investigated by n.m.r. spectroscopy. The chemical shifts of the pyrrole NH and Lactam protons of bilirubin and its dimethyl ester in DMSO indicate a strong interaction with the solvent. Inter-proton distances were calculated from nuclear Overhauser effects (NOE), selective and non-selective relaxation times (T1) and rotational correlation times taken from 13C relaxation times. The interproton distances indicate that the conformation of the skeleton of bilirubin and its dimethyl ester in DMSO is similar to that of bilirubin and mesobilirubin in the crystalline state and in chloroform solutions, except for a possible slight twist of the pyrrolenone rings about the methine bonds, which may be a consequence of solvation of the NH groups by DMSO. Unlike in chloroform solutions, no direct hydrogen-bonding occurs between the carboxylic acid and the lactam groups of bilirubin in DMSO, as shown by the absence of an NOE between these groups. The fast exchange of the pyrrole NH protons with 2H shows that no hydrogen-bonding occurs between these protons and the propionic residues, in line with their solvation by DMSO. From the above results, and from the slowness of the internal motion of the propionic residues of bilirubin and its dimethyl ester, it is concluded that these residues are tied to the skeleton via bound solvent molecules. PMID- 6284121 TI - Kinetic studies on the reduction of cytochrome c. Reaction with dihydroxy conjugated compounds (catechols and quinols). AB - The kinetics of reduction of cytochrome c by catechol(s), quinol(s) and related compounds were investigated by stopped-flow spectrophotometry. Studies on the influence of pH on the rates indicate that only deprotonated forms of these compounds act as reducing agents, with the dianionic forms being the most effective. The pH-independent second-order rate constants are reported. Hammett treatment of the effects of substituents on the aromatic ring structure of the reductants show that for electron transfer to occur the charge on the deprotonated species must not be withdrawn on to the substituents. Possible sites for electron donation to cytochrome c are discussed, and the results indicate that the haem edge is a likely candidate. PMID- 6284125 TI - A fibronectin-binding glycoprotein from human platelet membranes. AB - Fibronectin ('cold-insoluble globulin') has been suggested as a possible mediator of platelet adhesion. A fibronectin-binding protein as partially purified from washed solubilized human platelet membranes by affinity chromatography on fibronectin-Sepharose. The isolated protein migrated as a single band on sodium dodecyl sulphate/polyacrylamide-gel electrophoresis with an Mr (relative molecular mass) of approx. 125 000 under reducing conditions. The protein migrated as a dimer in non-reduced gels. The purified protein did not react with immunoglobulins against fibrinogen or fibronectin when tested in crossed immunoelectrophoresis or electroimmunoassay. The protein and purified fibronectin formed a complex that had a significantly faster mobility in crossed immunoelectrophoresis than did native fibronectin. The presence of heparin in the binding-protein-fibronectin mixture resulted in an even faster mobility of the complex, whereas the mobility of native fibronectin was unaffected. Crossed affinoimmunoelectrophoresis of the complex using different lectins suggested that the binding protein is a glycoprotein containing N-acetylglucosamine residues. The complex, but not purified fibronectin, bound to phenyl-Sepharose on crossed hydrophobic-interaction immunoelectrophoresis. The results strongly suggest the presence of a fibronectin-binding glycoprotein in the platelet membrane. PMID- 6284126 TI - The stereochemical course of phosphoryl transfer catalysed by glucose 6 phosphatase. AB - Rat liver microsomal glucose 6-phosphatase catalyses phosphoryl transfer between D-glucose 6-[(R)-16O,17O,18O]phosphate and D-glucose with retention of configuration at the phosphorus atom. Since individual phosphoryl-transfer steps appear in general to occur with inversion of configuration, this observation is most simply interpreted in terms of a double-displacement mechanism with a phosphoryl-enzyme intermediate. Such an intermediate has been proposed previously from kinetic and 32P-labelling experiments. PMID- 6284127 TI - Biochemical heterogeneity of skeletal-muscle microsomal membranes. Membrane origin, membrane specificity and fibre types. AB - 1. Microsomes were isolated from rabbit fast-twitch and slow-twitch muscle and were separated into heavy and light fractions by centrifugation in a linear (0.3 2m) sucrose density gradient. The membrane origin of microsomal vesicles was investigated by studying biochemical markers of the sarcoplasmic-reticulum membranes and of surface and T-tubular membranes, as well as their freeze fracture properties. 2. Polyacrylamide-gel electrophoresis showed differences in the Ca(2+)-dependent ATPase/calsequestrin ratio between heavy and light fractions, which were apparently consistent with their respective origin from cisternal and longitudinal sarcoplasmic reticulum, as well as unrelated differences, such as peptides specific to slow-muscle microsomes (mol.wts. 76000, 60000, 56000 and 45000). 3. Freeze-fracture electron microscopy of muscle microsomes demonstrated that vesicles truly derived from the sarcoplasmic reticulum, with an average density of 9nm particles on the concave face of about 3000/mum(2) for both fast and slow muscle, were admixed with vesicles with particle densities below 1000/mum(2). 4. As determined in the light fractions, the sarcoplasmic-reticulum vesicles accounted for 84% and 57% of the total number of microsomal vesicles, for fast and slow muscle respectively. These values agreed closely with the percentage values of Ca(2+)-dependent ATPase protein obtained by gel densitometry. 5. The T-tubular origin of vesicles with a smooth concave fracture face in slow-muscle microsomes is supported by their relative high content in total phospholipid and cholesterol, compared with the microsomes of fast muscle, and by other correlative data, such as the presence of (Na(+)+K(+))-dependent ATPase activity and of low amounts of Na(+)-dependent membrane phosphorylation. 6. Among intrinsic sarcoplasmic-reticulum membrane proteins, a proteolipid of mol.wt. 12000 is shown to be identical in the microsomes of both fast and slow muscle and the Ca(2+)-dependent ATPase to be antigenically and catalytically different, though electrophoretically homogeneous. 7. Basal Mg(2+)-activated ATPase activity was found to be high in light microsomes from slow muscle, but its identification with an enzyme different from the Ca(2+)-dependent ATPase is still not conclusive. 8. Enzyme proteins that are suggested to be specific to slow-muscle longitudinal sarcoplasmic reticulum are the flavoprotein NADH:cytochrome b(5) reductase (mol.wt. 32000), cytochrome b(5) (mol.wt. 17000) and the stearoyl-CoA desaturase, though essentially by criteria of plausibility. PMID- 6284128 TI - Inhibition of cerebral protein kinase activity and cyclic AMP-dependent ribosomal protein phosphorylation in experimental hyperphenylalaninaemia. AB - Studies were carried out to elucidate the mechanisms underlying the diminished phosphorylation of cerebral ribosomal protein in experimental hyperphenylalaninaemia [Roberts & Morelos (1980) Biochem. J.190, 405-419]. Administration of N(6),O(2)'-dibutyryl cyclic AMP or 3-isobutyl-1-methylxanthine, which increased phosphorylation of the S6 protein of cerebral 40S ribosomal subunits in control infant rats, did not counteract the decreased phosphorylation of this ribosomal protein resulting from intraperitoneal administration of a loading dose of l-phenylalanine. N(2),O(2)'-Dibutyryl cyclic GMP had no effect on cerebral ribosomal-protein phosphorylation in either control or hyperphenylalaninaemic animals. The phenylalanine-induced decrease in ribosomal protein phosphorylation was associated with decreased protein kinase activity in cerebral cytosolic and microsomal preparations. However, the maximal protein kinase response to cyclic AMP added in vitro was unaltered by prior administration of phenylalanine in vivo. The heat-stable protein inhibitor of cyclic AMP-dependent protein kinases decreased the activity of these enzymes by about 90% and eliminated the phenylalanine-induced difference in protein kinase activity in the absence of added cyclic AMP. Intracisternal administration of doses of dibutyryl cyclic AMP or 3-isobutyl-1-methylxanthine which increased the cyclic AMP-dependent protein kinase activity ratio in control infant rats was without effect on this index in phenylalanine-treated animals. Dibutyryl cyclic GMP had no effect on the protein kinase activity ratio in either group of animals. These results suggest that inhibition of cerebral cyclic AMP-dependent protein kinases by abnormally high concentrations of phenylalanine may contribute to the decrease in cerebral ribosomal-protein phosphorylation in experimental hyperphenylalaninaemia. PMID- 6284129 TI - Calcium ions modulate hormonally stimulated progesterone production in isolated ovarian cells. AB - Swine granulosa-luteal cells incubated in Ca2+-deficient medium (5 muM final Ca2+ concentration) for short time periods produced diminished quantities of progesterone in response to lutropin. Maximally stimulating effects of prostaglandin E2 and L-adrenaline were also impaired significantly. Diminished progesterone production could not be attributed to alterations in protein synthesis or cell viability. Under Ca2+-deprived conditions, the stimulatory actions of cholera toxin, 3-isobutyl-1-methylxanthine and 8-bromo cyclic AMP were also significantly impeded. Administration of a presumptive antagonist of transmembrane Ca2+ influx (verapamil) or of EGTA to chelate extracellular Ca2+, significantly decreased the total cellular content of Ca2+, and antagonized the actions of lutropin. Micromolar concentrations of trifluoperazine mimicked the suppressive effects of Ca2+ deprivation. Conversely, the bivalent-cation ionophore, ionophore A23187, significantly augmented the stimulation of progesterone produced by lutropin. Thus the present observations implicate Ca2+ in the modulation of hormonally stimulated progesterone production in isolated ovarian cells, and suggest that Ca2+ may influence one or more processes distal to, or independent of, cyclic AMP generation. In addition, the susceptibility of progesterone biosynthesis to inhibition by trifluoperazine suggests a possible role for calmodulin in the ovary. PMID- 6284131 TI - Properties of ubiquinol oxidase reconstituted from ubiquinol-cytochrome c reductase, cytochrome c and cytochrome c oxidase. AB - Ubiquinol-cytochrome c reductase (Complex III), cytochrome c and cytochrome c oxidase can be combined to reconstitute antimycin-sensitive ubiquinol oxidase activity. In 25 mM-acetate/Tris, pH 7.8, cytochrome c binds at high-affinity sites (KD = 0.1 microM) and low-affinity sites (KD approx. 10 microM). Quinol oxidase activity is 50% of maximal activity when cytochrome c is bound to only 25% of the high affinity sites. The other 50% of activity seems to be due to cytochrome c bound at low-affinity sites. Reconstitution in the presence of soya bean phospholipids prevents aggregation of cytochrome c oxidase and gives rise to much higher rates of quinol oxidase. The cytochrome c dependence was unaltered. Antimycin curves have the same shape regardless of lipid/protein ratio, Complex III/cytochrome c oxidase ratio or cytochrome c concentration. Proposals on the nature of the interaction between Complex III, cytochrome c and cytochrome c oxidase are considered in the light of these results. PMID- 6284130 TI - Differential sensitization to deoxyribonuclease I of Xenopus vitellogenin and albumin genes during primary and secondary induction of vitellogenesis by oestradiol. AB - The sensitivity to DNAase (deoxyribonuclease) I (which preferentially digests transcribed sequences) of vitellogenin and albumin genes in liver and erythrocytes of male Xenopus after primary and secondary induction of vitellogenesis by oestrogen was measured by hybridization to cDNA (complementary DNA) of the residual DNA after enzymic digestion of isolated nuclei. Vitellogenin sequences were rendered selectively more sensitive to limited DNAase-I digestion (15-20% of DNA rendered acid-soluble) during primary hormonal activation (5 days) of vitellogenin genes in liver, but not erythrocyte, nuclei. Hormone withdrawal (25 days after first injection) did not result in reversion to a pre-activation gene configuration, nor did secondary hormonal stimulation (5 days after second and 25 days after first injection) augment the sensitivity of the genes to digestion by the nuclease. Similar hormone treatment did not affect the sensitivity of the constitutively expressed albumin genes in liver nuclei, nor their insensitivity in erythrocyte nuclei. Under the same conditions, globin genes remained indigestible in liver nuclei. It is concluded that primary induction of vitellogenesis in male Xenopus liver is accompanied by relatively long-lasting (3-4 weeks) change in the configuration of vitellogenin genes in hepatic nuclei which is not reversed or further modified during short-term oestrogen withdrawal or upon secondary stimulation. PMID- 6284132 TI - Effects of cross-linked dimers of ribonuclease A or of lysozyme on the processing of endocytosed peroxidase by hepatoma cells. AB - Cross-linked dimers of ribonuclease, added at a concentration of 0.05 mg/ml to the culture medium of hepatoma (HTC) cells, were previously shown to inhibit intracellular degradation of peroxidase taken up by endocytosis. Intracellular localization showed that endocytosed peroxidase does not reach lysosomes in dimer treated cells. The present study shows that preloading of lysosomes with fluorescent anti-peroxidase IgG, obtained by exposing HTC cells for 48 h to 0.1 mg of antibody/ml, restores intracellular degradation of endocytosed peroxidase. Moreover, accumulation of peroxidase into lysosomes, which no longer occurs in dimer-treated cells, occurs again under these conditions. We conclude that inhibition of transfer of peroxidase from phagosomes to lysosomes is most likely to be the alteration resulting from the exposure of the cells to ribonuclease dimer, rather than inhibition of fusion between phagosomes and lysosomes. The dimer of another basic protein, lysozyme added at a concentration of 0.2 mg/ml to the culture medium, is shown to induce the same type of effects as does the dimer of ribonuclease; the half-life of endocytosed peroxidase increased from 5 to 15 h after 2 h exposure of HTC cells to dimerized lysozyme. The effect of both dimers on intracellular protein processing can be reversed by addition of 100 mm galactose to the culture medium, up to 5 h after pretreatment of the cells. The dimers of ribonuclease A or of lysozyme have thus probably the same mechanism of action. Evidence that the two dimers share the same binding sites on the cells is presented. PMID- 6284133 TI - Evidence that poly(ADP-ribose) polymerase is involved in the loss of NAD from cultured rat liver cells. AB - Rat hepatocytes cultured from 24h lose 60% of their NAD content. By using the differential response to inhibitors of the two major enzymes that catabolize NAD in mammalian cells, it is shown that poly(ADP-ribose) polymerase is responsible for the loss of NAD. The relevance of this observation to the use of cultured hepatocytes for the study of DNA repair induced by carcinogens is discussed. PMID- 6284134 TI - Regulation of neutral cholesterol esterase and acyl-CoA : cholesterol acyltransferase in the rat adrenal gland. AB - The activities of neutral cholesterol esterase and acyl-CoA : cholesterol acyltransferase in rat adrenal gland were measured at various time intervals over 24 h. The activity of cholesterol esterase displayed diurnal rhythm, with a major peak at the onset of darkness coinciding with the peak in the diurnal rhythm of plasma corticosterone concentration. The activity of acyl-CoA : cholesterol acyltransferase also exhibited a characteristic diurnal rhythm, with the minimum activity occurring 3 h after the onset of darkness. The profile of the rhythm exhibited by the activity of the esterifying enzyme was similar to the mirror image of the pattern of diurnal rhythm in the activity of 3-hydroxy-3 methylglutaryl-CoA reductase. Microsomal non-esterified cholesterol showed a gradual decline with a significant decrease in concentration at the onset of darkness, thus suggesting that diurnal removal of cholesterol in the environment of the esterifying enzyme and hydroxymethylglutaryl-CoA reductase leads to such diurnal decrease or increase in the activities of these two enzymes. Acute administration of corticotropin led to a 3-fold increase in the activity of cholesterol esterase, a 50% decrease in the activity of acyl-CoA : cholesterol acyltransferase and a 2-fold increase in the activity of hydroxymethylglutaryl CoA reductase. Corticotropin administration also resulted in a significant decrease in microsomal non-esterified cholesterol and increase in plasma corticosterone concentration. These observations suggest that corticotropin plays an important part in generating the diurnal rhythm in the activities of the three enzymes. PMID- 6284135 TI - No major thermogenic role for (Na+ + K+)-dependent adenosine triphosphatase apparent in hepatocytes from hyperthyroid rats. AB - (Na+ + K+)-dependent ATPase activity, heat production and oxygen consumption were increased by 59%, 62% and 75% respectively in hepatocytes from tri-iodothyronine treated rats. Ouabain at concentrations of 1 and 10 mM decreased oxygen uptake by 2--8% in hepatocytes from euthyroid rats and by 5--15% in hepatocytes from hyperthyroid animals. Heat output was decreased by 4--9% with the glycoside in isolated liver parenchymal cells from the control animals and by 11% in the cells from the tri-iodothyronine-treated animals. These results do not support the hypothesis that hepatic (Na+ + K+)-ATPase plays a major role in increased heat production in hepatocytes from hyperthyroid rats. PMID- 6284136 TI - Desensitization of tumour Leydig cells by lutropin: evidence for uncoupling of the lutropin receptor from the guanine nucleotide-binding protein. AB - Purified rat Leydig tumour cells were pretreated with lutropin and the effect on the subsequent response to lutropin was determined. Maximal cyclic AMP production was achieved with the same concentration of lutropin in control and lutropin pretreated cells; however, the maximum stimulated level in pretreated cells was only 30% of controls. The sensitivity to lutropin was decreased in lutropin pretreated cells [ED(50) (dose that produces a response that is 50% of the maximum response) 60+/-5.7ng/ml and 8+/-1.8ng/ml (mean+/-s.d., n=3) for controls], as was the rate of maximal cyclic AMP production (0.58, compared with 1.89pmol/10(6) cells per min for controls). However, cholera-toxin-stimulated cyclic AMP production was not decreased by lutropin pretreatment, and a potentiation was seen at all time points studied (up to 6h). Pre-incubation with lutropin caused a decrease in specific (125)I-labelled human choriogonadotropin binding; however, this decrease was abolished if the cells were washed under acidic conditions (pH3.0 for 2min at 4 degrees C), indicating that occupation but not loss of the lutropin receptors had taken place. The effect of pretreating the cells with lutropin on adenylate cyclase activity in purified plasma membranes was also investigated. In plasma membranes from control cells both guanosine 5' [beta,gamma-imido]triphosphate [p(NH)ppG] plus lutropin and NaF plus lutropin caused a 50-60-fold linear increase in cyclic AMP production over 40min compared with 15-fold with p(NH)ppG and 6-fold with lutropin alone. In plasma membranes isolated from lutropin-treated cells the NaF-plus-lutropin- and the p(NH)ppG stimulated cyclic AMP production rates were unchanged but no effect of lutropin could be demonstrated with or without added p(NH)ppG. In contrast the plasma membranes from dibutyryl cyclic AMP-treated cells had similar cyclic AMP production rates to control cells with all stimulants studied. The present evidence obtained from studies both with intact cells and with isolated plasma membranes indicates that the initial lutropin-induced desensitization of the rat Leydig tumour cell is due to a lesion in the hormone-receptor coupling to the guanine nucleotide regulatory protein. This process is apparently not mediated by cyclic AMP. PMID- 6284137 TI - A comparison of polyamine metabolism in normal and transformed baby-hamster kidney cells. AB - Transformed baby-hamster-kidney cells contain higher intracellular concentrations of polyamines than do normal cells. The difference is greater in high-density confluent cultures. Transformed cells incorporate exogenous putrescine into the cells at a faster rate than do normal cells. They also show a marked increase in the rate of spermine biosynthesis compared with normal cells. Transformed cells grown to high cell densities released about 10% of their polyamines into the culture medium in a non-specific manner. In contrast, normal cells, under the same culture conditions, release up to 50% of their intracellular polyamines into the medium almost exclusively as free or conjugated spermidine. The elevated levels of polyamines found in transformed cells therefore appear to be the result of altered transport of polyamines across the cell membrane and of increased rates of biosynthesis. PMID- 6284139 TI - Involvement of peroxide and superoxide in the oxidation of hemoglobin by nitrite. PMID- 6284138 TI - Is Percoll innocuous to cells? AB - Peritoneal macrophages from mice, isolated rat liver Kupffer cells and rat testis Leydig cells ingested large numbers of Percoll particles, a gradient medium widely used for separation of cells and subcellular organelles by density gradient centrifugation. A decrease in the percentage of macrophages adhering to plastic also occurred after exposure of the cells to Percoll, even at 4 degrees C, a temperature at which Percoll was not ingested. The effect of Percoll on macrophage adherence may involve a loose association between the density medium and the cell surface. Other cell-surface-related phenomena may also be affected by prior exposure of cells to Percoll. PMID- 6284141 TI - Ionic channels of some glycine-rich synthetic polypeptides. PMID- 6284140 TI - Regulation of nuclear uptake of "activated" receptor-glucocorticoid complex by pyrophosphate. PMID- 6284142 TI - Photoactivated cross-linking of prolactin to hepatic membrane binding sites. PMID- 6284143 TI - An electron spin resonance study of the novel radical cation produced during the horseradish peroxidase-catalyzed oxidation of tetramethylhydrazine. PMID- 6284144 TI - Reversible dissociation of triiodothyronine-nuclear receptor complexes by mercurial and chaotropic reagents. PMID- 6284145 TI - Increased central alpha-2 adrenergic receptors measured with [3H] yohimbine in the presence of sodium ion and guanylnucleotides. PMID- 6284146 TI - Developmental changes in the form of the Arrhenius plots of rat liver plasma membrane adenylate cyclase and 5'-nucleotidase activities. PMID- 6284147 TI - An enzyme that catalyzes hydrolysis of fructose-2, 6-bisphosphate. PMID- 6284148 TI - Apparent phosphorylation - dephosphorylation of soluble phosphatidic acid phosphatase in rat liver. PMID- 6284149 TI - hCG increased phosphorylation of proteins in primary cultures of porcine Leydig cells. PMID- 6284150 TI - Mn2+ does not uncouple adenosine "Ra" receptors from the liver adenylate cyclase. PMID- 6284151 TI - The EGF receptor-kinase has multiple phosphorylation sites. PMID- 6284152 TI - Role of lipids in gonadotropin releasing hormone agonists and antagonist binding to rat pituitary. PMID- 6284153 TI - Involvement of poly (ADP-ribose) metabolism in induction of differentiation of HL 60 promyelocytic leukemia cells. PMID- 6284154 TI - Intracellular O2 gradients in cardiac myocytes. Lack of a role for myoglobin in facilitation of intracellular O2 diffusion. PMID- 6284155 TI - Purification and characterization of cytochrome C3 (Mr 26,000) isolated form Desulfovibrio desulfuricans Norway strain. PMID- 6284157 TI - Identification and characterization of angiotensin II receptors in cardiac sarcolemma. PMID- 6284156 TI - Evidence that lipid peroxidation in microsomal membranes of epidermis is associated with generation of hydrogen peroxide and singlet oxygen. PMID- 6284158 TI - DCCD inhibits proton translocation and electron flow at the second site of the mitochondrial respiratory chain. PMID- 6284159 TI - Evidence that 19-hydroxyandrostenedione is secreted by the adrenal cortex and is under the control of ACTH and the renin-angiotensin system in man. PMID- 6284160 TI - Study of the protein-ubiquinone interaction in succinate-cytochrome C reductase with azido-ubiquinone derivatives. PMID- 6284161 TI - Oxidative metabolism of hydralazine. Evidence for nitrogen centered radicals formation. PMID- 6284162 TI - TSH-treatment of thyroid slices increases the amount of DNA released from nuclei by mild DNase-I digestion. PMID- 6284163 TI - The induction of cytochrome P-448 dependent benzo(a)pyrene hydroxylase in Saccharomyces cerevisiae. PMID- 6284164 TI - Lectin binding of solubilized opiate receptors: evidence for their glycoprotein nature. PMID- 6284165 TI - Interaction of calcium antagonists with cyclic AMP phosphodiesterases and calmodulin. PMID- 6284166 TI - Isolation and characterization of a fragment of rat thyroglobulin gene. PMID- 6284167 TI - FBJ virus-induced osteosarcoma has type V collagen consisting of A, B and C-like chains in addition to type I collagen. PMID- 6284168 TI - Secretion by mononuclear phagocytes of lysosomal hydrolases bearing ligands for the mannose-6-phosphate receptor system of fibroblasts: evidence for a second mechanism of spontaneous secretion? PMID- 6284169 TI - The sites on the regulatory component of adenylate cyclase which are ADP ribosylated by cholera toxin. PMID- 6284170 TI - Formation of the 11,12-diol as a metabolite of benzo(a)pyrene by rat skin in vivo. PMID- 6284171 TI - Identification of the regulatory subunit of a cAMP-dependent protein kinase in Dictyostelium discoideum. PMID- 6284172 TI - In vivo and in vitro effects of tetrahydroisoquinolines and other alkaloids on rat pituitary function. AB - Several tetrahydroisoquinolines (TIQs) were tested for their in vitro and in vivo capacities to modulate prolactin (PRl) and beta-endorphin (beta-end) secretion by the rat pituitary and for their abilities to displace [3H]spiroperidol and [3H]naloxone binding from pituitary and hypothalamic membranes. Receptor binding studies showed that TIQs could be classified as having (a) higher affinity for opiate receptors (tetrahydropapaverine, papaverine, 6-methylsalolinol, 1 carboxysalsolinol and 3',4'-deoxy-norlaudanosolinecarboxylic acid), (b) higher affinity for the dopamine receptor (salsolinol and 7-methylsalsolinol), or (c) approximately equal affinity for the two binding sites (6,7-dimethylsalsolinol and tetrahydropapaveroline, THP). In freely moving male rats, THP produced a several-fold increase in plasma PRL levels. This effect was not altered by co administration of naloxone but was attenuated by dopamine. In vitro several TIQs reversed the inhibitory effect of dopamine on PRL secretion by cultured anterior pituitary cells. The order of potencies of the TIQs in this system paralleled their order of potencies in the dopamine receptor assay. THP, the most potent dopamine antagonist, also blocked dopamine-mediated inhibition of beta-endorphin secretion from neurointermediate lobe cells in culture. These data demonstrate that THP and some other TIQs can act as dopamine antagonists in radioreceptor assays, in cell culture and in vivo. PMID- 6284173 TI - Possible role of calcium uptake and calmodulin in adrenal glomerulosa cells: effects of verapamil and trifluoperazine. AB - The effects of verapamil and trifluoperazine were examined on isolated rat adrenal glomerulosa cells so as to assess the role of calcium ion influx and calmodulin in the function of this cell population. Verapamil (10(-5) and 10(-4) moles/1) slightly reduced the basal production rate of aldosterone and strongly inhibited the response to angiotensin II, potassium ions, corticotrophin (ACTH) and dibutyryl cyclic AMP (db-cAMP). The concentration of verapamil required to reduce the response to these agonists by 50% varied between 2 and 6 mumoles/1. Trifluoperazine (30 mumoles/1) slightly increased the basal production rate of aldosterone. The response to angiotensin and potassium was variably antagonized by 3 mumoles/1 trifluoperazine and completely inhibited by the drug at 30 mumoles/1. The antagonist at a concentration of 3 mumoles/1 exerted either a facilitatory or inhibitory effect on the response to ACTH and db-cAMP, depending on the concentration of the agonist. Trifluoperazine at a concentration of 30 mumoles/1 reduced the response to both agonists to a level which was 2-3 fold higher than that observed in appropriate control samples. The present results indicate that (1) calcium influx is an essential event in the aldosterone stimulating action of angiotensin II, potassium ions, ACTH and cyclic AMP; (2) stimulation by angiotensin II and potassium ions are completely dependent on calmodulin; (3) stimulation by ACTH and cyclic AMP is mediated by calmodulin dependent and independent mechanisms. PMID- 6284174 TI - Selective inhibition of a cyclic nucleotide independent protein kinase (G type casein kinase) by quercetin and related polyphenols. AB - The effect of quercetin and a number of structurally related phenolic compounds upon the activity of three different purified protein kinases was examined. Whereas the catalytic subunit of a cyclic AMP-dependent protein kinase and an A type (using only ATP) cyclic nucleotide-independent casein kinase (CKA) were not affected, a G type (using GTP as well as ATP) casein kinase (CKG) was selectively inhibited by several bioflavonoid structures. Kinetic studies showed that quercetin behaved as a competitive inhibitor toward the nucleotidic substrate and exhibited a high affinity for the ATP (Ki = 0.75 microM) and GTP (Ki = 0.22 microM) site of the enzyme. Considering the CKG inhibitory potency of a series of flavonoid, cinnamic acid and coumarin derivatives, it is suggested that the biological activity lays upon a common structural feature involving a phenolic ring bearing a side chain with conjugated double bonds and an oxygenated function, as found in the coumaroyl residue. These observations suggest that quercetin and related compounds may lead to a shift in intracellular protein phosphorylations by selectively inhibiting a particular type of protein kinase activity (CKG). It remains to be established whether this process may contribute to the mechanism of action of flavonoids upon cellular metabolism, particularly in the case of malignant cells. PMID- 6284175 TI - Effects of steroids on beta-adrenergic binding sites in sheep pineal glands. AB - As an initial step in investigations of putative differences between central nervous system light-sensitive mechanisms in seasonally shedding and non-shedding breeds of sheep, some beta-adrenoceptor characteristics of Merino sheep pineal glands were determined, using [3H]dihydroalprenolol as the labelled ligand. Overall, a dissociation constant of 17.2 +/- 2.6 nmoles/l and a daytime beta receptor density of 1.6 +/- 0.3 pmoles/mg were determined at 37 degrees. The binding sites exhibited stereospecificity, saturability and apparent homogeneity. 17 beta-Estradiol and progesterone implants that provided hormone concentrations in the physiological range had no significant effect on pineal beta-receptors in male sheep castrated shortly after birth. Dexamethasone injections, on the other hand, in doses sufficient to loosen the attachment of wool fibres to the skin, resulted in decreased pineal beta-receptor density and increased receptor affinity for dihydroalprenolol. This effect was apparently not mediated by altered plasma catecholamine concentrations, since the glucocorticoid treatment did not affect jugular venous noradrenaline, adrenaline or dopamine levels. The possible involvement of glucocorticoids in the regulation of wool growth could thus have a central neuronal component, medicated via action on pineal beta adrenoceptors in sheep; however, the existence of the putative gonadal steroid feedback on beta-adrenoceptor-mediated pineal function remains to be demonstrated in this species. PMID- 6284176 TI - Inhibition by etazolate (SQ 20009) and cartazolate (SQ 65396) of adenosine stimulated [3H]cAMP formation in [2-3H]adenine-prelabeled vesicles prepared from guinea pig cerebral cortex. PMID- 6284177 TI - Resistance of angiotensin I converting enzyme to hydrolysis by serine proteases. PMID- 6284178 TI - Role of phospholipids in the inhibitory action of DDT and permethrin on the nerve ATPase of lobster, Homarus americanus. AB - Efforts were made to understand the nature of the site of 1,1-bis-(p chlorophenyl)-2,2,2-trichloroethane (DDT) inhibition of nerve ATPase. THe phospholipid content of nerve preparations from the walking leg of the lobster was reduced by treating them with phospholipase A, or with a chloroform-methanol mixture at -75 degrees. By these treatments the enzymes lost approximately 70 or 95% of their phospholipids and 50-80% of their Na,K- and Ca-ATPase activities. The lost ATPase activities could be partially restored by the addition of phospholipids, either the ones extracted from the lobster nerves or those from commercial sources. ATPase inhibition by DDT and permethrin was found to be highest in preparations where the phospholipids were removed by the above treatments, next highest with the untreated original enzyme, and least with the reconstituted ATPase regardless of the source of phospholipids used for reconstitution. This tendency was more pronounced in the case of Ca-ATPase. The effects of DDT and permethrin on inhibition of reconstituted Ca-ATPase were higher when the insecticide was first added to the protein portion and the enzyme was then reconstituted with the phospholipids, than when the same amount of insecticide was first added to the phospholipids which were then used for reconstitution. PMID- 6284179 TI - Cytotaxin-induced cAMP peak in granulocytes: its relationship to crawling movements, chemokinesis and chemotaxis. AB - The relationship between the short transient intracellular increase in cAMP levels on the one hand and chemotaxis or crawling movements on the other hand was investigated using human and equine granulocytes. C5ades arg, f-met-leu-phe, human serum albumin and immunoglobulin were used as stimulating agents. There was no strict correlation between the induction of crawling movements or of chemokinesis in general and the generation of the cAMP peak. But there was so far a strict parallelism between the occurrence of the chemotactic response and the cAMP peak. However, the magnitude of the peak was not representative for the extent of directional locomotion. It seems possible that cAMP is an essential step in the transduction of the chemotactic response, but there is no proof for a causal relationship as yet. PMID- 6284180 TI - Epinephrine inhibits protein synthesis in isolated mouse hepatocytes through alpha adrenergic receptors in a calcium dependent way. PMID- 6284181 TI - [Detection of rotavirus in hospitalized and ambulatory infants with acute diarrhea in Santiago de Chile]. PMID- 6284182 TI - [Whooping cough-like syndrome in malnourished patients]. PMID- 6284183 TI - [Adrenocorticotropic hormone in hypertension]. AB - The corticotropic function of the adenohypophysis was investigated in patients with essential hypertension considering the stage of the disease and development of crisis. There was found functional activation of the system under study at the labile stage of essential hypertension without crisis as well as in patients with crisis at labile and stable stages of the disease. Unidirectional shifts were observed in the function of the vasopressor part of the hypothalamus neurohypophyseal system. PMID- 6284184 TI - Purified glycoproteins of Sendai virus envelope affect human erythrocyte membrane fluidity. PMID- 6284185 TI - [Kinetic characteristics of microsomal NAD-glycohydrolase natural and solubilized with a non-ionic surface-active substance]. AB - Microsomal rat spleen NAD-glycohydrolase was solubilized by Nonidet P40. The solubilized enzyme shows Nicotinamide inhibition and pH dependence at the same extent as unsolubilized microsomal one. It differs from the latter in having a higher affinity for NAD and NADP, and in showing two peaks, instead of one, on electrofocusing: the former with a pH 5 pI without any activity, the latter with a pH 4, 1 pI with a high NAD-ase activity. PMID- 6284186 TI - [Microneurography (author's transl)]. PMID- 6284187 TI - [Fine structure of the locus coeruleus in the mouse (author's transl)]. AB - The aim of this study is to clarify the fine structural organization of the mouse locus coeruleus (LC), which has not been thoroughly investigated yet, by a light and electron microscope. In a light microscopic study we used the wet fluorescent method and counted numbers of fluorescent LC cells and obtained 1326 +/- 64 in oneside, and many processes of LC cells reached and expanded into the latero dorsal tegmental nucleus and nucl. parabrachialis medialis and lateralis. By EM study of glyoxylic acid and permanganate fixed material, LC cells (the principal cells, (16.8 +/- 2.9) X (27.8 +/- 5.1) micrometers), which contained small and large cored vesicles characteristic for NA, were divided into 2 types. The one was rich in Golgi apparatuses, ER and other cell organelles, the other being poor in these cell organelles. Another type of cells, devoid of cored vesicles, were small in size (12 X 15 micrometers) and poor in cell organelles. In the neuropil of LC, the % of axon terminals containing NA cored vesicles amounted to 2.2 (+/- 0.67) % of all axon terminals (a total of 10,545). Counted NA terminals (a total of 232) showed apposition to dendrites at 43%, free endings at 52%, contacts to LC cells at 5%. Moderate number of dendrites containing cored vesicles contacted with LC cell bodies or their dendrites especially in the caudal part of LC. PMID- 6284188 TI - Diagnostic pitfalls. The pre-auricular swelling. PMID- 6284189 TI - Sodium fluoride activation of human lymphocyte adenylate cyclase is reduced in old age. PMID- 6284190 TI - A phase I study of misonidazole and pelvic irradiation in patients with carcinoma of cervix. AB - A Phase I study of oral daily misonidazole (MISO) with conventional pelvic irradiation, has been conducted in patients with carcinoma of the cervix Stages IB, IIB, IIIB and IVA. MISO was administered in daily dosages to sequential groups of patients at doses of 0.15 g/m2, 0.30 g/m2 or 0.45 g/m2 for 22 days over 5 weeks. Sixteen patients were assigned to each dose level. Using a double-blind randomization, they received either placebo (3/16) or MISO (13/16). The major dose-limiting toxicity was peripheral neuropathy (PN). None of the 13 patients receiving 0.15 g/m2 or the 13 receiving 0.3 g/m2 developed PN. However, 6/13 at the 0.45 g/m2 level (total dose less than or equal to 9.9 g/m2) developed PN. Additional patients were entered at this level and a total of 13/26 developed PN, which was considered of clinically significant severity in 9. Symptoms of PN have persisted from 1 week to 10 months, and have been completely reversed in 9/13 patients. Pharmacological parameters were examined for correlation with clinically evident toxicities. Although peak plasma MISO levels and half-lives did not correlate significantly with PN, there was a significant correlation between the calculated "area under the curve" (AUC) and PN. No correlation exists between PN and total urinary excretion of MISO or the O-demethylation product. A daily dose of 0.45 g/m2; MISO (total dose less than or equal to 9.9 g/m2) is considered to produce an acceptable level of toxicity for this patient population. PMID- 6284192 TI - Comparative studies of transformed hamster cells by Herpesvirus hominis types 1 and 2. AB - Hamster embryo cells transformed by Herpesvirus Type 2 (HSV-2) virus (two separate cell lines) and by HSV-1 virus (one cell line) were inoculated into hamsters. The morphological appearance of tumours was studied by both light and electron microscopy, and the results compared to similar studies of tumours induced by SV40 virus. The tumours were also tested for immunogenicity, and for susceptibility to contact suppression with BCG. The results indicated that the tumours induced by inoculation of HSV-transformed cells were distinct from those of other DNA viruses; that the two cell lines transformed by HSV-2 were very similar but distinct from the cell line transformed by HSV-1; that all 3 HSV tumour cell lines were weakly immunogenic, and 2 were susceptible to contact inhibition with BCG. The relevance of the morphological and immunogenic properties of the animal tumour is discussed with relation to the possible importance of the findings to human malignant disease. PMID- 6284191 TI - Dietary wheaten bran in baboons: long-term effect on the morphology of the digestive tract and aorta, and on tissue mineral concentrations. AB - Two groups of 13 young baboons, each consisting of 8 males and 8 females, were fed on either high- or low-bran diets (based on wheat of either high or low extraction rate) for a period of 26 months. All animals grew well and remained in good condition throughout. Male (but not female) baboons on the high-bran diet had lower (p less than 0.05) concentrations of zinc in serum and bone, despite a low phytate: zinc molar ratio and a high intake of zinc. Particle-induced X-ray emission analysis showed there to be lower concentrations of calcium, copper, zinc, sulphur, potassium and nickel in the livers of baboons on the high-bran diet (P less than 0.005). Baboons on the low-bran diet passed smaller quantities of softer faeces, they had fewer nodules of lymphoid tissue in the distal portion of the colon (P less than 0.05), and within mucosal microherniations of their ileo-caecal valves the epithelial cells showed a greater tendency to squamous transformation (P less than 0.05). Mucosal microherniations of the ileo-caecal valves tended to be more frequent and larger in size (P less than 0.05) in animals of the low-bran group. Morphometrical studies did not reveal any differences in the general structural development of the digestive tract. The high-bran diet had no effect on serum cholesterol concentrations, nor on the incidence or severity of atherosclerotic lesions of the aorta. PMID- 6284193 TI - Tumorigenicity and tumour-graft rejection of polyoma virus-transformed fibroblast T-lymphocyte hybrids. AB - In anticipation of the use of functional T-lymphocyte hybrids in adoptive immunotherapy, the differentiation and tumorigenicity of hybrid clones generated by fusion of a T lymphocyte derived from F1 (DBA/J2 x AKR) mouse spleen, and a polyoma virus-transformed fibroblast initiated from C3H mouse cells, were studied. The hybrid cells grew in suspension and had an appearance (by transmission and scanning electron microscopy) very similar to that of the lymphocytic line. The hybrid and the different clones could induce tumour grafts. Malignancy was dominant in newborn mice where tumours were obtained in all mouse strains (allogeneic or semi-allogeneic) inoculated. In adult mice, the hybrid cells were tumorigenic in C3H and F1 (DBA/J2 x AKR), whereas there was complete tumour rejection in allogeneic (C57/BL6) or semi-allogeneic (DBA/J2 and AKR) mice. The role played by major histocompatibility antigens in the graft rejection is discussed. The histology of the tumour grafts was intermediate between fibrosarcoma and lymphosarcoma. PMID- 6284195 TI - The influence of cyclic nucleotides on DNA synthesis in skin. AB - Total saline-soluble protein, total DNA, and 3H-thymidine labelling of DNA were measured in cultured guinea-pig skin during a 3-day incubation period. An early rise in total soluble protein and a fall in total DNA indicated loss of the cells damaged in cutting the skin samples. Samples incubated with dibutyryl cyclic adenosine monophosphate (db cAMP) showed an early stimulation of DNA synthesis. Cyclic guanosine monophosphate (cGMP) stimulation of DNA synthesis showed a peak after 48 h incubation, probably reflecting a slower rate of entry of the compound into the epidermal cells. PMID- 6284194 TI - Serum inhibitory effect on myeloperoxidase in human cancer. AB - The myeloperoxidase level is strongly depressed in leucocytes from cancer patients (-59%). Moreover sera of the same subjects induce a significant decrease in the myeloperoxidase activity of healthy subjects' leucocytes (-43%). We have found no convincing evidence of any correlation between the level of the inhibitory effect of sera and the tumour type. The fact that sera from cancer patients interfere with neutrophil responses in this way does not exclude the possibility that abnormal cellular functions were also operative. PMID- 6284197 TI - Independent binding sites in mouse 5.8S ribosomal ribonucleic acid for 28S ribosomal ribonucleic acid. AB - Limited digestion of mouse 5.8S ribosomal RNA (rRNA) with RNase T2 generates 5'- and 3'-terminal "half-molecules". These fragments are capable of independently and specifically binding to 28S rRNA, so there exist at least two contacts in the 5.8S rRNA for the 28S rRNA. The dissociation constants for the 5.8S/28S, 5' 5.8S fragment/28S, and 3' 5.8S fragment/28S complexes are 9 x 10(-8) M, 6 x 10(-8) M, and 13 x 10(-8) M, respectively. Thus, each of the fragment binding sites contributes about equally to the overall binding energy of the 5.8S/28S rRNA complex, and the binding sites act independently, rather than cooperatively. The dissociation constants suggest that the 5.8S rRNA termini from short, irregular helices with 28S rRNA. Thermal denaturation data on complexes containing 28S rRNA and each of the half-molecules of 5.8S rRNA indicate that the 5'-terminal binding site(s) exist(s) in a single conformation while the 3'-terminal site exhibits two conformational alternatives. The functional significance of the different conformational states is presently indeterminate, but the possibility they may represent alternative forms of a conformational switch operative during ribosome function is discussed. PMID- 6284196 TI - An increase in the Na+/K+-ATPase activity of erythrocyte membranes in workers employed in a lead refining factory. AB - To clarify the relationship between erythrocyte Na+/K+-ATPase activity and haematological findings, several clinical laboratory examinations were performed on 31 male workers employed in a scrap lead refining factory and, as controls, 50 male workers employed in railway construction. The results were: (1) Values for erythrocyte Na+/K+-ATPase activity, blood and urine lead, urine delta aminolaevulinic acid, and urine coproporphyrin of lead workers were significantly higher than those of the controls (p less than 0.01). (2) A strongly positive relationship between blood lead and erythrocyte Na/K-ATPase activity was observed in lead workers (r = 0.473, p less than 0.01). (3) A strongly negative relationship between Na+/K+-ATPase activity and intracellular sodium was observed in both groups (lead workers; r = -0.601, p less than 0.01: controls; r = 0.595, p less than 0.01). PMID- 6284198 TI - Rotational mobility of an erythrocyte membrane integral protein band 3 in dimyristoylphosphatidylcholine reconstituted vesicles and effect of binding of cytoskeletal peripheral proteins. AB - Band 3 protein was isolated from human erythrocyte membranes, purified, and reconstituted into a well-defined phospholipid bilayer matrix (dimyristoylphosphatidylcholine). The preparation yielded uniform single bilayered vesicles of the diameter 40--80 nm. The rotational motion of band 3 was studied by saturation transfer electron spin resonance (ESR) spectroscopy of covalently attached maleimide spin-labels. The rotational mobility changed in response to the host lipid phase transition. The rotational correlation time was in a range from 73 (37 degrees C) to 94 microseconds (26 degrees C) in the fluid phase and from 240 (15 degrees C) to 420 microseconds (5 degrees C) in the solid phase. The motion was analyzed based on the anisotropic rotation of band 3 in the reconstituted vesicles. To obtain information on the rotational diffusion constant around the axis parallel to the membrane normal, we made an attempt to measure the angle between the spin-label magnetic axis and the membrane normal. The result gave 3.9 x 10(4) s-1 at 37 degrees C as a rough estimate for the diffusion constant. This is compatible to anisotropic rotation of a cylinder of radius 3.3 nm in a two-dimensional matrix with inner viscosity 2 P and inner thickness 4 nm. The cytoskeletal peripheral proteins caused a definite increase in the rotational correlation time (from 73 to 180 microseconds at 37 degrees C, for example). The restriction of the rotational mobility was shown to be due to the ankyrin-linked interaction between band 3 and spectrin-actin-band 4.1 proteins in the reconstituted membranes. PMID- 6284199 TI - Developmentally regulated nonhistone proteins: evidence for deoxyribonucleic acid binding role and localization near deoxyribonuclease I sensitive domains of precartilage cell chromatin. AB - Differentiation of cartilage from precartilage mesenchyme in the chick embryo is accompanied by the loss of two abundant nonhistone proteins (Mr 35 500 and 125 000) termed PCP 35.5 and PCP 125. Here we examine the distribution of these and other developmentally regulated nonhistones in nuclease-sensitive regions of precartilage and cartilage chromatin. In particular, we show that PCP 35.5 is a tight DNA-binding protein that is localized near deoxyribonuclease I (DNase I) sensitive regions of precartilage chromatin. Localization of nonhistones was demonstrated by excising domains of precartilage chromatin with DNase II which are simultaneously highly enriched in PCP 35.5, in PCP 125, and DNase I sensitive DNA sequences. These domains comprise at least 25% of the cell's DNase I sensitive sequences, as well as small DNase I resistant regions with which the two nonhistones are associated. These findings suggest that PCP 35.5 (and possibly PCP 125) may play a developmentally regulated role nearby DNase I sensitive domains of the cartilage progenitor cell chromatin. PMID- 6284200 TI - Histone H2A phosphorylation in animal cells: functional considerations. AB - We have sought to determine the role of histone H2A phosphorylation in chromatin by examining the distribution of the phosphorylated and unphosphorylated forms of this core histone within the nuclei of mouse and human cells. At any time, only about 15% of the H2A of whole chromatin is in the phosphorylated form, and its phosphate is rapidly turned over, even in quiescent cells that contain a functional nucleus. The phosphorylations and dephosphorylations are not specifically relate to progress through the cell cycle, nor to DNA synthesis or repair, and they are not selectively nucleolar. Euchromatin is substantially enriched with phosphorylated H2A but is not the exclusive repository of it. Possible roles of this modification of H2A are considered. PMID- 6284201 TI - Lipid environments in the yolk lipoprotein system. A spin-labeling study of the lipovitellin/phosvitin complex from Xenopus laevis. AB - Lipid/protein and lipid/lipid interactions in the yolk lipoprotein complex from Xenopus laevis were examined by introducing a series of lipid spin-labels into the complex and observing the electron spin resonance spectra as a function of the position of the label along the lipid chains, temperature, pH, and charge on the lipid polar head group. Analyses of the spectra show that, in addition to the expected component arising from lipid associated with protein, a second component with increased segmental flexibility and the greater temperature dependence characteristic of lipid/lipid interactions is observed. These spin-labeling data and supporting compositional data indicate that much of the lipid is organized into a lipid-rich region or pool, consistent with the earlier model derived from electron microscopy and diffraction data and with companion 31P and 2H nuclear magnetic resonance data reported in the preceding paper [Banaszak, L. J., & Seelig, J. (1982) Biochemistry (preceding paper in this issue)]. The bilayer-like component exhibits a greater restriction of motion compared to vesicles of the isolated lipids at the same temperature, as would be expected for a relatively small lipid pool. Phospholipids exchange between the two motionally distinguishable environments. The equilibrium binding undergoes a shift between these two environments as a function both of pH and of the charge on the phospholipid polar head group. This shift in average binding affinity is opposite in direction to that reported for membrane proteins and implicates negatively charged groups on the protein that repel negatively charged phospholipids. This effect is greatly reduced by alkaline phosphatase treatment, suggesting that some of the lipid binding sites are in close proximity to phosphorylated residues on the protein. PMID- 6284202 TI - DNA polymerase delta: one polypeptide, two activities. AB - DNA polymerase delta from rabbit bone marrow has an associated 3'-5'-exonuclease. Previous studies demonstrated a Stokes radius of 45.5 A by gel filtration and a sedimentation coefficient of 6.5 S by zone sedimentation. Thus, a molecular weight of 122000 and a frictional coefficient of 1.39 were calculated [Byrnes, J. J., & Black, V. L. (1978) Biochemistry 17, 4226-4231]. Several problems obstructed further purification and definition of DNA polymerase delta. The small amount of protein obtained limited further purification as the nonspecific loss of enzyme in subsequent procedures was excessive. Furthermore, the amount of protein recovered was insufficient for conventional analysis. These difficulties have been overcome, and DNA polymerase delta has been purified to apparent homogeneity. Under conditions of nondenaturing microgel electrophoresis, DNA polymerase b aggregates to molecular weight species of 300000 and higher. In situ assays for DNA polymerase and exonuclease in these gels generate concordant activity profiles. Upon sodium dodecyl sulfate gel electrophoresis, delta is a single polypeptide of 122000 apparent molecular weight. The DNA polymerase incorporates between 250000 and 300000 nmol of thymidine deoxyribonucleoside monophosphate (dTMP) into poly(dA)/oligo(dT) (mg of protein)-1 h-2 at 37 degrees C; the exonuclease simultaneously hydrolyzes 13% of the newly synthesized DNA. Aphidicolin, considered to be a specific inhibitor of DNA polymerase alpha, inhibits both the DNA polymerase and 3'-5'-exonuclease activities of delta. DNA polymerase alpha from rabbit bone marrow does not share a common subunit with delta. Therefore, aphidicolin binding is not specific for alpha, and conclusions based upon the supposition that it is must be reconsidered. PMID- 6284203 TI - Preparation of two-dimensional arrays from purified beef heart cytochrome c oxidase. AB - A method for preparing two-dimensional crystals from highly purified beef heart cytochrome c oxidase is described. This involves mixing the enzyme with phosphatidylcholine and then extracting excess lipid with deoxycholate. The reconstituted crystals show PI symmetry. Alternating rows of monomers are more closely packed (along the b axis) than in the previously described P121 crystals. However, the monomer structure in projection is the same in the two crystal forms. The P121 crystal form has been reacted with trypsin. This treatment did not alter the crystals but removed most of the impurities present in these cytochrome c oxidase preparation of low purity. PMID- 6284204 TI - An enzyme activity from Escherichia coli that attacks single-stranded deoxyribopolymers and single-stranded deoxyribonucleic acid containing apyrimidinic sites. AB - We have isolated an enzyme activity from extracts of Escherichia coli that catalyzes the hydrolysis of phosphodiester bonds in the single-stranded deoxyribopolymer (dU.[3H]dT)(2000) containing depyrimidinated sites created by enzymatic removal of uracil with uracil-DNA glycosylase. Nondepyrimidinated polymer is not degraded by the enzyme, nor is the depyrimidinated polymer degraded after reduction of apyrimidinic sites with sodium borohydride. The enzyme also degrades circular M13 DNA containing uracil or denatured phage PBS2 DNA (which naturally contains uracil) only after the removal of uracil from these substrates by uracil-DNA glycosylase. Undamaged or depurinated duplex PM2 or ColE1 DNA and undamaged M13 single-stranded DNA are not attacked by the enzyme. The activity sediments in glycerol gradients with a relative s value of 4.2 S and has a Stokes radius of 31 A, yielding a calculated Mr approximately 56,000. The fractional ratio (f/f0) is calculated at 1.25. The enzyme has no requirement for any known cofactors and is insensitive to inhibition by p (chloromercuri)benzoate. Activity is inhibited in the presence of adenosine 5'0 triphosphate (ATP), tRNA or high ionic strength and is slightly stimulated by MnCl2 or CaCl2. PMID- 6284205 TI - Cytochrome c oxidase binding of hydrogen peroxide. AB - Oxidized cytochrome c oxidase can bind hydrogen peroxide, as evidenced by changes in its spectrum and its ability to use hydrogen peroxide as an electron acceptor in cytochrome c oxidation. The affinity of the oxidized enzyme for hydrogen peroxide is high, with a Kd of less than 10 microM, and the binding is inhibited by ligands of cytochrome a3. Oxidized cytochrome c oxidase, in submitochondrial particles or solubilized in several ionic and nonionic detergents, binds peroxide with comparable affinities. The size of the spectral shift observed upon peroxide binding depends on the pH of the solution and differs in extinction coefficient between preparations, but all preparations tested appeared to bind peroxide. The differences in the magnitude of the spectral shift upon peroxide binding to different preparations suggest that oxidized cytochrome c oxidase as prepared may be made up of more than one species and that the proportion of the species which binds peroxide varies with the preparation. These studies of the binding of peroxide clarify the mechanism by which cytochrome c oxidase catalyzes the reduction of oxygen to water without the formation of free-radical intermediates. PMID- 6284206 TI - Determination of the distribution of sodium and potassium ion activated adenosinetriphosphatase among the various oligomers formed in solutions of nonionic detergents. AB - Sodium and potassium ion activated adenosinetriphosphatase [(Na+ + K+)-ATPase] can be dispersed from the membrane-bound state, with the stable retention of the capacity to display (Na+ + K+)-ATPase activity, by treatment with solutions of a homogeneous, nonionic detergent, octaethylene glycol dodecyl ether. The dispersed enzyme is incapable of turnover, however, in solutions where the free detergent concentration is above the critical micelle concentration. Treatment of solutions of this enzyme with the crosslinking reagent glutaraldehyde results in the quantitative, covalent coupling of the alpha-and beta-polypeptides. The various covalent products formed, when visualized on sodium dodecyl sulfate polyacrylamide gels, are integral oligomers of the asymmetric unit (alpha beta) of the enzyme. The noncovalent oligomers from which these products are derived can be separated on sucrose gradients based on differences in their respective sedimentation coefficients, but these sedimentation coefficients are highly dependent on the concentration of detergent in the gradient. Furthermore, the cross-linking assay reveals that changes in the aggregation state of the enzyme occur as detergent:protein ratios are varied or when the enzyme is added to the ATPase assay. These observations suggest that earlier conclusions about the oligomers of this enzyme present in detergent solution were significantly in error. PMID- 6284207 TI - Increased level of translatable collagenase messenger ribonucleic acid in rabbit synovial fibroblasts treated with phorbol myristate acetate or crystals of monosodium urate monohydrate. AB - We studied mechanisms governing production of the neutral proteinase collagenase by synovial cells. We used a model system of monolayer cultures of rabbit synovial fibroblasts stimulated to produce collagenase by treatment with phorbol myristate acetate or crystals of monosodium urate monohydrate. mRNAs from these and untreated cells were translated in a wheat germ cell-free system. Collagenase was not present in the culture medium or in the in vitro translation products of mRNA from untreated cells but was present in both the medium and translation products of stimulated cells, as analyzed by gel electrophoresis and immunoprecipitation with monospecific antibody. Induction of collagenase was prevented by treatment of the cells with alpha-amanitin (2 mug/ mL), an inhibitor of mRNA synthesis. We have concluded that the induction of collagenase synthesis by either phorbol myristate acetate or urate crystals is due to an increased level of translatable mRNA. PMID- 6284208 TI - Both parental deoxyribonucleic acid strands at each replication fork of replicating simian virus 40 chromosomes are cut by a single-strand-specific endonuclease. AB - We have measured the relative accessibility to a single-strand-specific endonuclease of the single-stranded DNA on the leading and lagging sides of replication forks in replicating simian virus 40 (SV40) chromosomes. To do this we have digested replicating SV40 chromosomes with a single-strand-specific endonuclease (P1 nuclease) and then characterized the intermediate and final products of digestion by sucrose gradient sedimentation and agarose gel electrophoresis. P1 nuclease rapidly and specifically cleaves parental DNA strands at replication forks, yielding intermediate and final cleavage products which are consistent with an approximately equal rate of nuclease cleavage on both sides of the fork. Thus, single-stranded DNA is approximately as accessible to P1 nuclease on the leading side of the fork as on the lagging side; the simplest interpretation of this observation is that the stretch of single stranded DNA on the leading side is as long as that on the lagging side. PMID- 6284209 TI - Calcium-independent stimulation of Bordetella pertussis adenylate cyclase by calmodulin. AB - Bordetella pertussis produces an extracellular adenylate cyclase activity that is present in the culture medium of exponentially growing cells. We have determined that calmodulin (CaM) stimulates this enzyme both in the presence and in the absence of free Ca2+. In the presence of 90 micron Ca2+ half-maximal stimulation of the enzyme occurred at 95 pM calmodulin. Comparable levels of calmodulin stimulation were observed when free Ca2+ levels were minimized by using EGTA containing buffers. However, the concentration of CaM required for half-maximal stimulation of B. pertussis adenylate cyclase in the presence of 1 nM free Ca2+ was 24 nM. The apparent affinity of the enzyme for calmodulin was also significantly enhanced by Mn2+. In addition, troponin I inhibited calmodulin stimulation of the bacterial adenylate cyclase. Photoaffinity cross-linking experiments using azido[125I]calmodulin and B. pertussis adenylate cyclase revealed only one major cross-linked product having a molecular weight of 97000. It is proposed that the catalytic subunit of the calmodulin-sensitive adenylate cyclase is 77000. PMID- 6284210 TI - Activation of skeletal muscle phosphorylase phosphatase. Effects of proteolysis and divalent cations. PMID- 6284211 TI - Identification of the glucagon receptor by covalent labeling with a radiolabeled photoreactive glucagon analogue. AB - The photoreactive 125I-labeled glucagon-NAPS [125I-labeled 2-[2-nitro-4 azidophenyl)sulfenyl]-Trp25-glucagon] was used to label the glucagon receptor sites in rat liver plasma membranes. The proteins labeled were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis with or without reduction with dithiothreitol. The photoaffinity peptide specifically labeled a number of bands with apparent molecular weights greater than 200000 and probably at least two protein bands in the molecular weight range 52000-70000. The relative amounts of radioactivity associated with these bands and their relative mobilities differed in samples from reduced and unreduced membranes. Their relative mobilities also differed with percent acrylamide cross-linking, suggesting a glycoprotein nature and the presence of intramolecular disulfide bonds. A nonspecifically labeled band with an apparent molecular weight of 27000-28000 also displayed a similar behavior. Photolabeling in the presence of 0.1 mM guanosine 5'-triphosphate (GTP) decreased the amount of radiolabeling of these bands, suggesting their involvement in the glucagon stimulation of adenylate cyclase. The photolabeled receptor in the membranes, solubilized with Lubrol-PX and fractionated on an Ultrogel AcA22 column, eluted with an apparent molecular weight of 200000-250000. Addition of GTP to the solubilized glucagon receptor of nonirradiated membranes caused complete dissociation of the complex. Gel electrophoresis of the partially purified radiolabeled receptor identified the same protein components observed in photolabeled membranes. These results indicate that the glucagon receptor is an oligomer probably composed of at least two different subunits that are linked together or greatly stabilized by disulfide bonds. They also show that 125I labeled glucagon-NAPS can be used effectively to covalently label the putative glucagon receptor and thus aid in its further characterization. PMID- 6284212 TI - Extended X-ray absorption fine structure studies of cytochromes c: structural aspects of oxidation-reduction. AB - EXAFS fluorescence spectra were recorded for high-potential c-type cytochromes which range in oxidation-reduction potential from +145 to +365 mV. No average Fe ligand bond length differences greater than 0.03 A were observed, for any cytochrome source of oxidation state. Least-squares analysis in combination with model calculations allowed limits to be set on the average Fe-N bond length (1.97 1.99 A) and the Fe-S bond length (2.29-2.32 A). A change of 0.05 A in either the average Fe-N or the Fe-S bond length is readily detectable with the range and quality of the data presented here. Two major conclusions are drawn from this study: In octahedrally coordinated iron porphyrin systems, Fe-N and Fe-S bond lengths are independent of oxidation-reduction potential, and they are also independent of oxidation state. A model for the regulation of oxidation-reduction potential in cytochrome c is proposed. PMID- 6284213 TI - Tumor viruses and early mouse embryos. PMID- 6284214 TI - Expression of adenoviral genetic information in productively infected cells. PMID- 6284216 TI - Metabolic co-operation between mammalian cells in culture. PMID- 6284215 TI - Surface glycoproteins of malignant human leukocytes. PMID- 6284217 TI - The reaction of cytochrome aa3 with (porphyrin) cytochrome c as studied by pulse radiolysis. AB - (1) Using the pulse-radiolysis and stopped-flow techniques, the reactions of iron free (porphyrin) cytochrome c and native cytochrome c with cytochrome aa3 were investigated. The porphyrin cytochrome c anion radical (generated by reduction of porphyrin cytochrome c by the hydrated electron) can transfer its electron to cytochrome aa3. The bimolecular rate constant for this reaction is 2 x 10(7) M-1 . s-1 (5 mM potassium phosphate, 0.5% Tween 20, pH 7.0, 20 degrees C). (2) The ionic strength dependence of the cytochrome c-cytochrome aa3 interaction was measured in the ionic strength range between 40 and 120 mM. At ionic strengths below 30 mM, a cytochrome c-cytochrome aa3 complex is formed in which cytochrome c is no longer reducible by the hydrated electron. A method is described by which the contributions of electrostatic forces to the reaction rate can be determined. (3) Using the stopped-flow technique, the effect of the dielectric constant (epsilon) of the reaction medium on the reaction of cytochrome C with cytochrome aa3 was investigated. With increasing epsilon the second-order rate constant decreased. PMID- 6284218 TI - Oxidation-reduction potentials of respiratory chain components in Thiobacillus A2. AB - (1) Cells of Thiobacillus A2 grown chemoautotrophically on thiosulfate or heterotrophically on succinate with oxygen contained b-, c-, o-, a- and a3-type cytochromes. The amount of cytochrome per mg of cell protein was much greater in thiosulfate-grown cells and differences in the relative concentrations of cytochromes were observed for the different growth conditions. (2) The half reduction potentials at pH 7.0 (Em,7.0) and spectral maxima of c-, b-, a- and a3 type cytochromes were similar in cells grown aerobically with thiosulfate or with succinate as the growth substrate. (3) The half-reduction potential of the 'invisible', or high-potential copper, as determined from the potentiometric behavior of the carbon monoxide-reduced cytochrome a3 complex at pH 8.0, was 365 mV. (4) Reducing equivalents from thiosulfate appear to enter the respiratory chain at the cytochrome c level; however, studies in cell-free extracts were limited due to a loss in respiratory activity with thiosulfate as a substrate upon cell disruption. PMID- 6284219 TI - The inhibitory effect of N,N'-dicyclohexylcarbodiimide in active sugar uptake by Rhodotorula glutinis. PMID- 6284220 TI - Transferrin receptors and iron uptake during erythroid cell development. AB - Experiments were performed to determine the level of transferrin receptors and rate of transferrin-bound iron uptake by various immature erythroid cell populations. Developing erythroid cells from the rat and mouse foetal liver at various stages of gestation were studied. In addition Friend leukaemic cells grown in culture were examined. The transferrin receptor level of Friend cells was similar to that of erythroid cells from the mouse foetal liver. During erythroid cell development the transferrin receptor level increased from about 300,000 per cell at the early normoblast stage to reach a maximum of about 8000,000 per cell on intermediate normoblasts. Further maturation of intermediate normoblasts was accompanied by a decline in the number of transferrin receptors, reaching a level of 105,000 in the circulating reticulocyte. The rate of iron uptake from transferrin during erythroid cell development was found to correlate closely with the number of transferrin receptors. In each of the immature erythroid cell populations studied the rate of iron uptake was about 36 iron atoms per receptor per hour. These results indicate that the level of transferrin receptors may be the major factor which determines the rate of iron uptake during erythroid cell development. PMID- 6284221 TI - Effect of adenylate cyclase activators and Mg2+ on the binding and the electron spin resonance spectra of N-methylmaleimide nitroxide in membrane particles from the liver fluke Fasciola hepatica. AB - Optimal conditions for activation of adenylate cyclase in membrane particles were studied. Enzyme activation with serotonin (5-hydroxytryptamine). NaF, and guanosine 5'-(3-O-thio)-triphosphate (GTP gamma S) was time-and temperature dependent. Mg2+ was required for enzyme activation. Adenylate cyclase that was activated by NaF or GTP gamma S was gradually inhibited by N-methylmaleimide while enzyme activated with serotonin and GTP responded faster to inhibition by the same sulfhydryl reagent. Th enzyme responded in a similar fashion to a spin labeled N-methylmaleimide analog 3-(maleimidomethyl)-2,2,5,5-tetramethyl-1 pyrolidinyloxyl (i.e., N-methylmaleimide nitroxide). Binding of the spin label was enhanced following enzyme activation by serotonin, NaF, or GTP gamma S in the presence of Mg2+. Activation of the enzyme was accompanied by an increase in the strong immobilization peaks in the EPR spectra. Both effects, the increase in binding and in the strong immobilization peaks, can be induced by Mg2+ alone. The results indicate that a general conformational induced by Mg2+ may be essential for adenylate cyclase activation. PMID- 6284222 TI - Low-affinity Na+ sites on (Na+ +K+)-ATPase modulate inhibition of Na+-ATPase activity by vanadate. AB - Na+-ATPase activity is extremely sensitive to inhibition by vanadate at low Na+ concentrations where Na+ occupies only high-affinity activation sites. Na+ occupies low-affinity activation sites to reverse inhibition of Na+-ATPase and (Na+, K+)-ATPase activities by vanadate. This effect of Na+ is competitive with respect to both vanadate and Mg2+. The apparent affinity of the enzyme for vanadate is markedly increased by K+. The principal effect of K+ may be to displace Na+ from the low-affinity sites at which it activates Na+-ATPase activity. PMID- 6284223 TI - Effects of ouabain and osmolarity on bumetanide-sensitive potassium transport in simian virus-transformed 3T3 cells. AB - Unidirectional potassium influx in simian virus-transformed 3T3 cells was dissected into a ouabain-inhibitable "pump' component, a bumetanide-sensitive and chloride-dependent "cotransport' component, and a residual "leak' flux. The bumetanide-sensitive component was stimulated 2-3-fold by a 60-min preincubation with ouabain. Subsequent washing of the cells and incubation in ouabain-free saline reversed both the inhibition of the Na+ pump and the stimulation of bumetanide-sensitive flux. Bumetanide-sensitive potassium influx was also stimulated by hypertonic cell shrinkage (induced by 0.1 M or 0.2 M sorbitol). This latter observation suggests that the bumetanide-sensitive system may play a role in cellular volume regulation. PMID- 6284224 TI - Removal of percoll from microsomal vesicles by gel filtration on sephacryl-S-1000 superfine. AB - A microsomal vesicle fraction was prepared from rat liver homogenate by centrifugation in gradients of Percoll. The microsomes were subjected to gel filtration on Sephacryl S-1000 Superfine, which resolved the microsomes from Percoll. The elution pattern of the microsomal marker enzyme NADPH-cytochrome c reductase showed that the main part of the enzyme was present in a peak at Kav about 0.1, while Percoll eluted in a broad peak at Kav about 0.7. The total yield of eluted enzyme activity was 85%. The gel filtration had to be carried out in the presence of 10 mM tris or NaCl. At lower ionic strength or in 0.25 M sucrose alone, anomalous behaviour of the Percoll particles and microsomes on the gel was observed. Electron microscopy of samples from the void volume fraction of the Sephacryl S-1000 Superfine column showed an almost complete removal of Percoll from the microsomes. Furthermore, the vesicle preparation was essentially free of membrane fragments. PMID- 6284225 TI - Inhibition of rat brain microsomal (Na+ + K+)-ATPase and K+-p nitrophenylphosphatase by periodic acid. AB - The effects of mild periodate exposure on the kinetics of (Na+ + K+)-ATPase and K+-p-nitrophenylphosphatase were studied using rat cerebral microsome preparations. Fifty percent inhibition of both enzyme activities was attained near 3 microM periodate concentrations. This inhibition was biphasic with time. Mg2+-ATPase and Mg2+-p-nitrophenylphosphatase activities were much less inhibited by periodate. Periodate inhibition was partially reversed by dimercaprol and dithiothreitol but not by diffusion. The possible reaction products formic acid, formaldehyde, glyceraldehyde, and acetaldehyde had no inhibitory effects in similar concentrations. Periodate exposure produced no detectable changes in the activation of (Na+ + K+)-ATPase by Na+, K+, Mg2+, or ATP. Residues shared by both (Na+ + K+)-ATPase and K+-p-nitrophenylphosphatase are both critical to hydrolytic function and sensitive to mild oxidation by periodate. PMID- 6284226 TI - Solvent effects on ouabain binding to the (Na+,K+)-ATPase of rat brain. AB - The effects of the solvents deuterated water (2H2O) and dimethyl sulfoxide (Me2SO) on [3H]ouabain binding to (Na+,K+)-ATPase under different ligand conditions were examined. These solvents inhibited the type I ouabain binding to the enzyme (i.e., in the presence of Mg2+ + ATP + Na+). In contrast, both solvents stimulated type II (i.e., Mg2+ + Pi-, Mg2+-, or Mn2+-dependent) binding of the drug. The solvent effects were not due to pH changes in the reaction. However, pH did influence ouabain binding in a differential manner, depending on the ligands present. For example, changes in pH from 7.05 to 7.86 caused a drop in the rate of binding by about 15% in the presence of Mg2+ + Na+ + ATP, 75% in the Mg2+ + Pi system, and in the presence of Mn2+ an increase by 24% under similar conditions. Inhibitory or stimulatory effects of solvents were modified as various ligands, and their order of addition, were altered. Thus 2H2O inhibition of type I ouabain binding was dependent on Na+ concentration in the reaction and was reduced as Na+ was elevated. Contact of the enzyme with the Me2SO, prior to ligands for type I binding, resulted in a greater inhibition of ouabain binding than that when enzyme was exposed to Na+ + ATP first and then to Me2SO. Likewise, the stimulation of type II binding was greater when appropriate ligands acted on enzyme prior to addition of the solvent. Since Me2SO and 2H2O inhibit type I ouabain binding, it is proposed that this reaction is favored under conditions which promote loss of H2O, and E1 enzyme conformation; the stimulation of type II ouabain binding in the presence of the solvents suggests that this type of binding is favored under conditions which promote the presence of H2O at the active enzyme center and E2 enzyme conformation. This postulation of a role of H2O in modulating enzyme conformations and ouabain interaction with them is in concordance with previous observations. PMID- 6284227 TI - Exchange between inorganic phosphate and adenosine triphosphate in (Na+,K+) ATPase. AB - (Na+,K+)-ATPase is able to catalyze a continuous ATP in equilibrium Pi exchange in the presence of Na+ and in the absence of a transmembrane ionic gradient. At pH 7.6 the Na+ concentration required for half-maximal activity is 85 mM and at pH 5.1 it is 340 mM. In the presence of optimal Na+ concentration, the rate of exchange is maximal at pH 6.0 and varies with ADP and Pi concentration in the assay medium. ATP in equilibrium Pi exchange is inhibited by K+ and by ouabain. PMID- 6284228 TI - Na+ channels and amiloride-induced noise in the mammalian colon epithelium. AB - (1) The effects of the Na+-channel blocker, amiloride, on the short-circuit current carried by Na+ was studied with fluctuation analysis, in rabbit descending colon epithelium. (2) In the presence of mucosal amiloride, the power spectrum of the Na+-current noise showed a Lorentzian component. When the Na+ current was reduced by increasing the blocker concentrations, the Lorentzian plateau decreased and corner frequency increased. Macroscopic short-circuit current and current-noise data are evidence for a two-state mechanism of the blocker interaction with the Na+ channel. (3) On- and off-rate constants for the blocker-receptor reaction, single-channel currents and Na+-channel density were calculated at room temperature and at 37 degrees C. Also, the activation energy for the amiloride-receptor reaction was estimated. The microscopic parameters obtained for the Na+ channel in the colon were similar to those found for Na+ channels in other tight epithelia. PMID- 6284229 TI - Reduction of opioid binding in neuroblastoma x glioma cells grown in medium containing unsaturated fatty acids. AB - Neuroblastoma x glioma cells NG108-15 were cultured in lipid-free medium supplemented with fatty acids of various chain length and unsaturation. Binding of 3H-labelled [DAla2]-[DLeu5]-enkephalin by membranes of cells grown in saturation fatty acids of different chain length was not significantly different from that of the control On the other hand, a proportional decrease of binding capacity with no change in residual receptor affinity was noticed when cells were cultured in medium containing fatty acids of increasing unsaturation. This decrease was time dependent and reached a maximum at about 48 h. Binding of [3H]dihydromorphine and [3H]naloxone was similarly affected. In contrast, when membranes of cells grown in normal medium were preincubated up to 3 h with unsaturated fatty acid and tested for opioid binding, no significant reduction was observed. Examination of the fatty acid composition of phospholipid from cells grown in linolenate indicated that a significant alteration of the acyl composition has occurred. To evaluate the underlying cause of this type of inhibition, the effect of linolenic acid on cell growth and protein synthesis was examined. When cells were cultured in 100 microM of this fatty acid, both growth and protein synthesis were retarded by 28% and 19%, respectively. Since opiate receptors are proteineous in nature, a reduction of protein synthesis may partially account for the loss of opioid binding activity. On the other hand, an increase of membrane fluidity is known to affect a number of cellular functions, including ligand-receptor recognition. Whether this can offer a satisfactory explanation for our observations remains to be established. PMID- 6284230 TI - Phospholipid miscibility in ternary mixtures. AB - The gel to liquid-crystalline phase transition of aqueous dispersions of phospholipid mixtures was investigated by means of the repartition of the spin label 2,2,6,6-tetramethylpiperidine-I-oxyl between aqueous space and lipid hydrocarbon region. The dimyristoylphosphatidylcholine (DMPC)/dibehenoylphosphatidylcholine (DBPC) and dipalmitoylphosphatidylcholine (DPPC)/DBPC phase diagrams indicate gel phase immiscibility, whereas the distearoylphosphatidylcholine (DSPC)/DBPC phase diagram indicates non-ideal gel phase miscibility at low DBPC molar fractions. Aqueous dispersions of DMPC/DPPC/DBPC ternary mixtures show two distinct phase transitions, the first associated with the melting of a DMPC/DPPC phase and the second with the melting of a DBPC phase. Aqueous dispersions of DMPC/DSPC/DBPC ternary mixtures show to phase transitions at low DSPC molar fractions; the first is probably associated with the melting of a DMPC/DSPC phase, and the second with the melting of a DBPC/DSPC phase. At high DSPC molar fractions, only one phase transition is observed; this suggests that all the lipids are mixed in gel state membranes. PMID- 6284231 TI - Tetrodotoxin-sensitive protein in the extracts from excitable tissues. AB - The sodium permeability of liposomes preincubated with the soluble fraction of brain and heart muscle homogenates was increased veratrine. The veratrine increment was decreased by tetrodotoxin. The effect was specific for the extracts from excitable tissues. Bovine serum and soluble fraction of liver homogenate induced neither veratrine- nor tetrodotoxin-sensitivity of the liposomes. Treatment of the excitable tissue extracts by pronase and heat denaturation caused their complete inactivation. Tetrodotoxin-sensitive factor could be fractionated by ammonium sulfate precipitation and by DEAE-Servacel chromatography. On a column of Sephadex G-200 it was eluted with the void volume. It is suggested that the tetrodotoxin-sensitive factor is a protein which could be a soluble precursor of the voltage-dependent sodium channels. PMID- 6284233 TI - Sulphydryl groups of (Na+ + K+)-ATPase from rectal glands of Squalus acanthias. Detection of ligand-induced conformational changes. AB - 1. Modification of the Class II sulphydryl groups on the (Na+ + K+)-ATPase from rectal glands of Squalus acanthias with N-ethylmaleimide has been used to detect conformational changes in the protein. The rates of inactivation of the enzyme and the incorporation of N-ethylmaleimide depend on the ligands present in the incubation medium. With 150 mM K+ the rate of inactivation is largest (k1 = 1.73 mM-1 . min-1) and four SH groups per alpha-subunit are modified. The rate of inactivation in the presence of 150 mM Na+ is smaller (k1 = 1.08 mM-1 . min-1) but the incorporation of N-ethylmaleimide is the same as with K+. 2. ATP in micromolar concentrations protects the Class II groups in the presence of Na+ (k1 = 0.08 mM-1 . min-1 at saturating ATP) and the incorporation is drastically reduced. ATP in millimolar concentrations protects the Class II groups partially in the presence of K+ (k1 = 1.08 mM-1 . min-1) and three SH groups are labelled per alpha subunit. 3. The K+-dependent phosphatase is inhibited in parallel to the (Na+ + K+)-ATPase under all conditions, and the ligand-dependent incorporation of N-ethylmaleimide was on the alpha-subunit only. 4. It is shown that the difference between the Na+ and K+ conformations sensed with N ethylmaleimide depends on the pH of the incubation medium. At pH 6 there is a very small difference between the rates of inactivation in the presence of Na+ and K+, but at higher pH the difference increases. It is also shown that the rate of inactivation has a minimum at pH 6.9, which suggests that the conformation of the enzyme changes with pH. 5. Modification of the Class III groups with N ethylmaleimide--whereby the enzyme activity is reduced from about 16% to zero- shows that these groups are also sensitive to conformational changes. As with the Class II groups, ATP in micromolar concentrations protects in the presence of Na+ relative to Na+ or K+ alone. ATP in millimolar concentrations with K+ present increases the rate of inactivation relative to K+ alone, in contrast to the effect on the Class II groups. 6. Modification of the Class II groups with a maleimide spin label shows a difference between Class II groups labelled in the presence of Na+ (or K+) and Class II groups labelled in the presence of K + ATP, in agreement with the difference in incorporation of N-ethylmaleimide. The spectra suggest that the SH group protected by ATP in the presence of K+ is buried in the protein. 7. The results suggest that at least four different conformations of the (Na+ + K+)-ATPase can be sensed with N-ethylmaleimide: (i) a Na+ form of the enzyme with ATP bound to a high-affinity site (E1-Na-ATP); (ii) a Na+ form without ATP bound (E1-Na); (iii) a K+ form without ATP bound (E2-K); and (iv) an enzyme form with ATP bound to a low-affinity site in the presence of K+, probably and E1-K-ATP form. PMID- 6284232 TI - Sulphydryl groups of (Na+ + K+)-ATPase from rectal glands of Squalus acanthias. Titrations and classification. AB - 1. (Na+ + K+)-ATPase from rectal glands of Squalus acanthias contains 34 SH groups per mol (Mr 265000). 15 are located on the alpha subunit (Mr 106000) and two on the beta subunit (Mr 40000). The beta subunit also contains one disulphide bridge. 2. The reaction of (Na+ + K+)-ATPase with N-ethylmaleimide shows the existence of at least three classes of SH groups. Class I contains two SH groups on each alpha subunit and one on each beta subunit. Reaction of these groups with N-ethylmaleimide in the presence of 40% glycerol or sucrose does not alter the enzyme activity. Class II contains four SH groups on each alpha subunit, and the reaction of these groups with 0.1 mM N-ethylmaleimide in the presence of 150 mM K+ leads to an enzyme species with about 16% activity. The remaining enzyme activity can be completely abolished by reaction with 5-10 mM N-ethylmaleimide, indicating a third class of SH groups (Class III). This pattern of inactivation is different from that of the kidney enzyme, where only one class of SH groups essential to activity is observed. 3. It is also shown that N-ethylmaleimide and DTNB inactivate by reacting with the same Class II SH groups. 4. Spin-labelling of the (Na+ + K+)-ATPase with a maleimide derivative shows that Class II groups are mostly buried in the membrane, whereas Class I groups are more exposed. It is also shown that spin label bound to the Class I groups can monitor the difference between the Na+- and K+-forms of the enzyme. PMID- 6284234 TI - Calcium-induced oscillations in K+ conductance and membrane potential of human erythrocytes mediated by the ionophore A23187. AB - The time-dependence of ionophore A23187-induced changes in the conductance of the Ca2+-sensitive K+ channels of the human red cell has been monitored with ion specific electrodes. The membrane potential was reflected in CCCP-mediated pH changes in a buffer-free extracellular medium, and changes in extracellular K+ activity and electrode potential of an extracellular Ca2+-electrode were recorded. Within a narrow range of ionophore-mediated Ca2+ influx, the above mentioned parameters were found to oscillate when ionophore was added to a suspension of glucose-fed cells. The period of oscillation was about 2 min/cycle depending on ionophore concentration, and the amplitude of hyperpolarization was about 60 mV, corresponding to a maximal gK+ of the same magnitude as gCl-. Without CCCP present no oscillation in K+ conductance was observed. The Ca2+ affinity for the opening process was in the micromolar range. The closing of the K+ channels was a spontaneous process in that the depolarization was well under way before the Ca2+-ATPase-mediated Ca2+ net efflux started. Below the Ca2+ influx range for oscillations, no response was observed for up to 20 min after the addition of ionophore. Above the upper limit, a permanent hyperpolarization resulted with an extracellular K+ activity increasing monotonically as a function of time. In experiments with ATP-depleted cells, responses of the latter type ensued at all ionophore concentrations above the lower limit. Addition of surplus EGTA to suspensions of hyperpolarized cells restores the normal membrane potential in the case of glucose-fed cells, whereas the K+-channels in ATP depleted cells remained open. PMID- 6284235 TI - Isolation and characterization of plasma membranes from krebs II ascite cells using Percoll gradient. AB - 1. Plasma membranes were isolated from Krebs II ascite cells grown in the mouse. Cells were disrupted by nitrogen cavitation in an isotonic alkaline buffer containing magnesium and ATP. Isolation was performed in an alkaline-buffered self-generating gradient of Percoll with an angular rotor. At each step of the preparation, the pH appeared as the critical aspect of our procedure. 2. External membrane markers were concanavalin A and 5'-nucleotidase (EC 3.1.3.5). They reached a relative specific activity of 10, whereas this value was only of 0.7 for the endoplasmic reticulum marker, NADH dehydrogenase (EC 1.6.99.3). 3. Plasma membrane from 4 ml packed cells were isolated within 1 h after homogenization with good yield: 50% and 67% of total [3H]concanavalin A and 5'-nucleotidase, respectively, were recovered in the two plasma membrane fractions. 4. Electron microscopy examination showed the presence of vesicles of different sizes devoid of other structural contaminants. 5. Using the specific binding of concanavalin A to the external cell membrane, it was calculated that about 50% of the total cell phospholipid and 10% protein are located in the plasma membrane. Their sphingomyelin content is much higher than in the whole cell, in contrast to phosphatidylinositol, known as a more specific endoplasmic reticulum phospholipid. PMID- 6284236 TI - 13C-NMR spectroscopy of acetyltyrosyl-guanidinated horse heart cytochrome c. AB - The tyrosine residues of guanidinated horse heart cytochrome c have been specifically acetylated by reaction with N-[1-13C]acetylimidazole (90 atom%). Acetylation was monitored by 13C-NMR spectroscopy. The tyrosine residues were found to show widely varying reactivities ranging from one that is completely and exclusively acetylated at low reagent concentration (residue 67) to one that is acetylated only when the protein is unfolded (residue 97). Homogeneous derivatives were prepared containing one (either residue 67 or 97), three 48, 67 and 74), or four (residues 48, 67, 74 and 97) O-[1-13C]acetyl groups. 13C-NMR spectra of selected derivatives were obtained at pH 5.8, in the presence of cyanide ion, in the ferrous and ferric oxidation states, and after denaturation with 6M guanidine hydrochloride. The O-[1-13C]acetyltyrosyl resonances gave chemical shift values ranging from 171.8 to 176.0 ppm. These resonances were assigned to specific groups based on the known order of reactivity of the tyrosyl side chains toward N-acetylimidazole. The chemical shift of O-[1 13C]acetyltyrosyl 67 was found to be particularly sensitive to changes in protein structure. The proximity of this group to the heme makes it subject to distance dependent paramagnetic and ring current effects. Acetylation of tyrosyl 74 gives rise to a pH-dependent equilibrium between conformers in the ferric state and a conformation change in the ferrous state. Acetylation of this residue also leads to an absorbance decrease at 695 nm that can be related to the 13C-NMR-detected conformational equilibrium. Addition of cyanide ion abolished this equilibrium. PMID- 6284237 TI - Effects of dithiothreitol on insulin-sensitive phosphodiesterase in rat fat cells. AB - Dithiothreitol activates the low-Km membrane-bound cyclic AMP phosphodiesterase when incubated with the enzyme in a cell-free system. To investigate the mechanism of its activation, we studied the effect of protease inhibitors. Isolated fat cells obtained from Sprague-Dawley rats were incubated in Krebs Henseleit Hepes buffer, pH 7.4, at 37 degrees C with and without insulin (2 nM, 10 min). A crude microsomal fraction prepared by differential centrifugation was suspended in 0.25 M sucrose containing 10 mM Tes buffer, pH 7.5, with and without 2 mM dithiothreitol and protease inhibitors at 4 degrees C for 48 h. Dithiothreitol stimulated the phosphodiesterase, in a time-dependent manner. As little as 0.02 mM dithiothreitol activated the enzyme, and the maximally effective dose was 2-10 mM. Among the various protease inhibitors tested, antipain, leupeptin, chymostatin and E-64 were the most effective in preventing activation of the enzyme by dithiothreitol. Antipain also inhibited release of the enzyme from the bound fraction. These results suggest that activation of the low-Km phosphodiesterase by dithiothreitol may be provoked by stimulation of an endogenous thiol protease. PMID- 6284238 TI - An examination of the cyanide derivative of bovine superoxide dismutase with electron-nuclear double resonance. AB - The Cu(II) sites of native, azido- and cyano-derivatives of bovine superoxide dismutase (superoxide:superoxide oxidoreductase, EC 1.15.1.1) have been examined by electron-nuclear double resonance (ENDOR). The ENDOR spectrum of the native protein taken at the g parallel extreme shows resolved structure due to the directly coordinated N-atoms of the histidine ligands. These spectra are too complex for interpretation but suggest inequivalent coupling between the electronic spin and the four ligand N-atoms. By contrast, the azido protein reveals one type of nitrogen with well-resolved hyperfine and quadrupole splittings (Azz = 37.9 +/- 1 MHz, Pzz = 1.54 +/- 0.02 MHz), and the cyano from reveals one well-resolved set of nitrogen lines (Azz = 47.8 +/- 0.4 MHz, Pzz = 1.62 +/- 0.01 MHz) and one type of partially resolved nitrogen (Azz = 37.0 +/- 1 MHz). The cyano form also reveals a complex spectrum in the low-frequency domain (1-10 MHz). Through isotopic substitution and computer stimulation, the spectrum is shown to be a composite of the ENDOR from the remote imidazole nitrogens and the cyanide nitrogen. The component of the hyperfine constant perpendicular to the C14N bonds axis is A perpendicular N = 3.9 +/- 0.3 MHz and along the bond axis is A perpendicular N approximately equal to 5.7 MHz. The quadrupole interaction appears to be greatest along the CN axis with Qz'z' = 1.0 +/- 0.1 MHz and Qx'x'y'y' approximately 0. Based on an analysis of the hyperfine and quadrupole interactions seen at two extremes of the electron paramagnetic spectrum, we propose a square-planar arrangement of three imidazole nitrogen and one CN- carbon around the copper. Within this plane two imidazole nitrogens are strongly coupled and magnetically equivalent, the third is inequivalent (slightly weaker hyperfine interactions) and forms a trans relationship with the cyanide. This model is consistent with other observations on the cyano-derivative. PMID- 6284239 TI - Molecular properties and kinetic studies on sphingomyelinase of Bacillus cereus. AB - A sphingomyelinase of Bacillus cereus was purified to a homogeneous state (512 U/mg, 2200-fold) as indicated by SDS-polyacrylamide gel electrophoresis and the molecular weight (23,300) was determined by sedimentation equilibrium. The enzyme contained loosely-bound magnesium atom. The addition of Mg2+ accelerated the enzyme reaction regardless of substrates and their physical state. The addition of Ca2+ also accelerated the enzyme reaction slightly, when water-soluble substrates, i.e., 2-hexadecanoylamino-4-nitrophenylphosphorylcholine and p nitrophenylphosphorylcholine, were used as substrates. On the other hand, the addition of Ca2+ inhibited enzyme reaction when mixed micelles of either sphingomyelin and Triton X-100 or sodium deoxycholate were used. The surface charge on mixed micelles affected the enzyme reaction. When the mixed micelle of sphingomyelin and Triton X-100 was used as substrate, Ca2+ proved to be a competitive inhibitor against Mg2+, with a Ki value of 33 microM. On the other hand, when the mixed micelle of sphingomyelin and sodium deoxycholate was used as substrate, Ca2+ stimulated the enzyme reaction at lower concentration in the presence of a low concentration of Mg2+, although higher concentrations of Ca2+ were still inhibitory. In this case, added Ca2+ may be used as a substitute of Mg2+ to neutralize the negative charge on the mixed micelle, improving the accessibility of sphingomyelinase to the micellar substrate. A cationic detergent, cetyltrimethylammonium bromide, seemed to denature or inactivate the enzyme. PMID- 6284240 TI - Ether-linked lipids of Balb/c3T3, SV3T3 and concanavalin A-selected SV3T3 revertant cells. AB - Ether-linked lipids were analyzed in Balb/c3T3, SV3T3 and Concanavalin A-selected SV3T3 revertant cells. The three cell lines were found to contain significant quantities of alk-1-enyl- and alkyl-linked phosphatidylethanolamine (PE) and phosphatidylcholine (PC) and small amounts of alkyldiacylglycerols. Compared to 3T3 cells, SV3T3 cells contain a higher amount of alk-1-enyl-linked PC, while in SV3T3 revertant cells the concentrations of the various ether lipids are similar to those of 3T3 cells. The major difference in the composition of ether groups of SV3T3 cells, compared to 3T3 cells, is an increase of 18:0 accompanied by a decrease of 18:1 in the alk-1-enyl-linked PE and PC. Alk-1-enyl-linked PC of SV3T3 revertant cells also shows an increase of 18:0, while the decrease of 18:1 was not statistically significant. PMID- 6284241 TI - Uptake of [3H]cholesterol from low density lipoprotein by cultured human fibroblasts. AB - The uptake of [3H]cholesterol from low density lipoprotein (LDL) was studied in LDL receptor-positive and receptor-negative human fibroblasts. In both cell lines the uptake depended upon temperature, time of incubation and the concentration of LDL in the medium. Although the incorporation of 125I-labeled LDL was minimal after 2 h of incubation in the receptor-negative (homozygous familial hypercholesterolemia, FH) cells, the uptake of [3H]cholesterol was only slightly less than that of the receptor-positive (WI-38) cells. With longer periods of incubation, a larger difference in labeled cholesterol incorporation was observed; this appeared to be due to a continued accumulation of the steroid in the WI-38 cells. After 8 and 24 h of incubation, some of the [3H]cholesterol was present as the ester in the WI-38 cells, but not the FH cells. Modified (reduced and methylated) LDL did not enter WI-38 cells by the receptor-mediated pathway during 2 h of incubation, as indicated by 125I uptake. [3H]Cholesterol uptake, however, was not significantly different from modified and unmodified LDL. While experiments indicated that significant amounts of cholesterol moved rapidly from LDL to cultured cells with a dependence on time and LDL concentration, no increase in total cell cholesterol was detected in either cell line. FH cells contained less total cholesterol and had a higher 3H specific activity than the WI-38 cells. These data suggest that there may be important mechanisms in addition to the LDL pathway for the movement of lipids into cells. PMID- 6284242 TI - Postnatal development of palmitate oxidation and mitochondrial enzyme activities in rat cardiac and skeletal muscle. AB - 1. The palmitate oxidation rate was measured in intact diaphragm and m. flexor digitorum brevis and in whole homogenates of heart, diaphragm and m. quadriceps of developing rats between late foetal life and maturity. Activities of the mitochondrial enzymes cytochrome c oxidase and citrate synthase were also determined. 2. Immediately after birth the palmitate oxidation rate increases markedly in both intact diaphragm and m. flexor digitorum brevis and falls gradually after day 1 to adult values which are about 35% of those at birth. 3. The oxidation capacities of diaphragm and m. quadriceps, but especially of heart, increase steadily during development, starting before birth and reaching adult values at 15-20 days postnatally. The activities of the mitochondrial enzymes show a similar developmental pattern. 4. In heart the increase of oxidative capacity is the result of an increase of both mitochondrial content and mitochondrial activity. The mitochondrial contents of diaphragm and m. quadriceps, on the other hand, decrease with age and the increase of their oxidative capacities is due to a large rise of the mitochondrial activity. PMID- 6284243 TI - Utilization of disaturated and unsaturated phosphatidylcholine and diacylglycerols by cholinephosphotransferase in rat lung microsomes. AB - 1. Cholinephosphosphotransferase catalyzes the conversion of diacylglycerol and CDPcholine into phosphatidylcholine and CMP. Incubation of rat lung microsomes containing phosphatidyl[Me-14C]choline with CMP resulted in an increase in water soluble radioactivity, suggesting that also in rat lung microsomes the cholinephosphotransferase reaction is reversible. 2. Microsomes containing 14C labeled disaturated and 3H-labeled monoenoic phosphatidylcholine were prepared by incubation of these organelles with [1-14C]palmitate and [9,10-3H2]oleate in the presence of 1-palmitoyl-sn-glycero-3-phosphocholine, ATP, coenzyme A and MgCl2. Incubation of these microsomes with CMP resulted in an equal formation of 14C- and 3H-labeled diacylglycerols, indicating that disaturated and monoenoic phosphatidylcholines were used without preference by the backward reaction of the cholinephosphotransferase. When in a similar experiment the phosphatidylcholine was labeled with [9,10-3H2]palmitate and [1-14C]linoleate, somewhat more 14C- than 3H-labeled diacylglycerol was formed. 3. The backward reaction was used to generate membrane-bound mixtures of [1-14C]palmitate- and [9,10-3H2]oleate- or of [9,10-3H2]palmitate- and [1-14C]linoleate-labeled diacylglycerols. When the microsomes containing diacylglycerols were incubated with CDPcholine, both 3H- and 14C-labeled diacylglycerols were used for the formation of phosphatidylcholine, indicating that there is no absolute discrimination against disaturated diacylglycerols. This observation is in line with our previous findings and indicates that also the CDPcholine pathway may contribute to dipalmitoylphosphatidylcholine synthesis in lung. PMID- 6284244 TI - Studies on some molecular properties of cytosol 5'-nucleotidase from rat liver. AB - A cytosol 5'-nucleotidase was purified from rat liver. It appeared to be homogeneous on the criteria of polyacrylamide gel electrophoresis. We estimated the approximate molecular weight of the enzyme to be 200 000 and concluded that the enzyme most likely exists in the native state as a tetramer. Our results suggest that adenine nucleotides, which are activators of the enzyme, induce its conformational change. PMID- 6284245 TI - Cytosolic and nuclear binding proteins for 2,3,7,8-tetrachlorodibenzo-p-dioxin in the rat thymus. PMID- 6284246 TI - Intrinsic protein phosphorylation in synaptic plasma membrane fragments from the rat. General characteristics and migration behaviour on polyacrylamide gels of the main phosphate acceptors. AB - Preparations enriched in synaptic membrane fragments from rat cerebral cortex contain protein kinases which phosphorylate membrane proteins in reactions dependent on cAMP, Ca2+ (in the absence of presence of calmodulin) or independent of these factors. In the present work characteristics of the main phosphorylated acceptors were studied and compared with the results of other investigations. Apparent molecular weights were estimated by determining electrophoretic mobility on gels of different acrylamide concentration. Irregular migration behaviour was detected by measuring free mobilities from Ferguson plots. Certain phosphate acceptors were found to exhibit anomalously low free mobilities and it was concluded that estimates of molecular weight for these acceptors were unreliable. PMID- 6284248 TI - Cyclic nucleotide phosphodiesterase activity in midpiece and tail of buffalo spermatozoa and its role in sperm motility. PMID- 6284247 TI - Effects of prostaglandins F alpha on dog thyroid cyclic AMP level and function. AB - Prostaglandins F1 alpha and F2 alpha, at high concentrations (greater than or equal to 28 microM) enhanced cyclic AMP accumulation in dog thyroid slices. At lower concentrations, they inhibited the cyclic AMP accumulation induced by thyrotropin (TSH), prostaglandin E1, and cholera toxin. This effect was rapid in onset and of short duration, calcium-dependent and suppressed by methylxanthines. Prostaglandin F alpha also inhibited TSH-induced secretion and activated iodide binding to proteins. These characteristics are similar to those of carbamylcholine action, except that prostaglandins F did not enhance cyclic GMP accumulation. The effect of prostaglandin F alpha was not inhibited by atropine, phentolamine and adenosine deaminase and can therefore not be ascribed to an induced secretion of acetylcholine, norepinephrine or adenosine. It is suggested that prostaglandins F act by increasing influx of extracellular Ca2+. Arachidonic acid also inhibited the TSH-induced cyclic AMP accumulation. However this effect was specific for TSH, it was enhanced in the absence of calcium and was not inhibited by methylxanthines or by indomethacin at concentrations which completely block its conversion to prostaglandin F alpha. Arachidonic acid action is sustained. This suggests that arachidonic acid inhibits thyroid adenylate cyclase at the level of its TSH receptor and that this effect is not mediated by prostaglandin F alpha or any other cyclooxygenase product. PMID- 6284249 TI - Regulation of alpha-galactosidase synthesis in Saccharomyces cerevisiae and effect of cerulenin on the secretion of this enzyme. AB - Saccharomyces cerevisiae -136ts MEL10 (a thermosensitive mutant whose RNA synthesis is inhibited at 37 degrees C but is normal at 23 degrees C), when grown at 23 degrees C in the presence of galactose, melibiose or L-arabinose, these cells synthesize alpha-galactosidase mRNA. In the simultaneous presence of both galactose and glucose the transcription of alpha-galactosidase mRNA is blocked. Glucose also interferes with mRNA translation, but the degree of inhibition depends on concentration and time of addition of the hexose to induced cells. It has been found that the final concentration of alpha-galactosidase produced by induced cells when transferred at the non-permissive temperature (37 degrees C) is inversely proportional to the incubation time in glucose. Cerulenin inhibits lipid formation on growing yeasts, but protein synthesis and selective permeability are not affected. The antibiotic partially inhibits secretion of alpha-galactosidase with a parallel accumulation of this enzyme in membranous structures, specially at the level of the plasma membrane. Induction of alpha galactosidase or cerulenin addition to growing cells, results in changes in the polypeptide composition of the plasma membrane. PMID- 6284250 TI - Angiotensin I converting enzyme of rat intestinal and vascular surface membrane. AB - High levels of angiotensin I converting enzyme are present in rat intestinal mucosa and in intestinal arteries. Homogenates of both tissues were subfractionated and fractions enriched in vascular plasma membrane or intestinal brush border were prepared. The preparations were identified and their purities established by marker enzyme enrichment and/or electron microscopy. Converting enzyme activity was highly enriched on both the vascular plasma membrane and the intestinal brush border. Subsequently the properties of these membrane-bound enzymes were compared. Both surface membrane-bound enzymes were highly sensitive to inhibition by captopril (SQ 14225) and teprotide (SQ 20881). Similar to converting enzyme isolated from other sources, they were also inhibited by bradykinin, angiotensin I, EDTA and o-phenanthroline. Finally, both membrane bound enzymes were relatively resistant to activation by sonication, freezing and thawing or detergent. These results demonstrate significant similarities between surface membrane-bound converting enzyme from vascular and non-vascular sites. In addition, in view of the possible relationship of kinins and angiotensins to gastrointestinal function and blood flow, inhibition of gastrointestinal converting enzyme by captopril may affect some aspects of intestinal physiology. PMID- 6284251 TI - Preparation of an N-acetyl-octapeptide of cholecystokinin. The role of N acetylation in protecting the octapeptide from degradation by smooth muscle tissues. AB - The C-terminal octapeptide of cholecystokinin (CCK-8) was acetylated on its lone N-terminal amino group using acetic anhydride in N,N-dimethylformamide. The acetylated derivative (Ac-CCK-8) and unreacted CCK-8 were separated from acetic anhydride and other reaction products by fractionation on Sephadex LH-20. Final purification was by thin-layer isoelectric focusing in a pH 2.5--4.0 gradient. The immunochemical properties of the octapeptide were unaffected by acetylation as measured by radioimmunoassay. The N-acetylated-octapeptide was equally as effective as unmodified CCK-8 in producing concentration-dependent isometric tension development in isolated cat gallbladder strips. Acetylation did, however, protect CCK-8 from N-terminal degradation by soluble peptidases that eluted from gallbladder and other smooth muscle tissues of the cat. Unmodified CCK-8 was degraded rapidly in the presence of these tissues and in buffers previously exposed to the same tissues. In contrast, the Ac-CCK-8 was resistant to N terminal degradation under the same conditions. Degradation of CCK-8 from its N terminus produces biologically inactive derivatives and could adversely affect in vitro studies. Since the acetylated-CCK-8 retained full biological and immunological activity, its use would eliminate the effect of extracellular proteolysis on CCK-8 action. PMID- 6284253 TI - [Absence of a magnetic field effect on Na+,K+-dependent ATPase]. PMID- 6284252 TI - [Electron spin resonance of phosphorylating and non-phosphorylating submitochondrial particles]. AB - Paramagnetic centres related with the work of ATP-synthetase, found earlier on mitochondria were studied on phosphorylating and non-phosphorylating submitochondrial particles (SMP). A complex doublet signal with half-width 38 . 10(-4) T was shown to be recorded only on actively phosphorylating and oligomycin inhibited SMP. It was found that the signal components were conditioned by different paramagnetic centres, one of which (g = 2.03) seemed to be the metal centre, more probable non-hemeiron. The nature of the second signal characterized by evenly saturating doublet components with g = 2.03 and g = 2.00 is unclear. Studies of flavin signal saturation have also shown that it consists of two components: one of them--saturating, specific for non-phosphorylating SMP, another--non-saturating, mostly observed in phosphorylating SMP, probably conditioned by flavin--the constituent of ATP-synthetase. PMID- 6284254 TI - [Effect of intracellular cyclic nucleotide injection on ionic current under voltage clamp conditions]. PMID- 6284255 TI - Effect of temperature and ions on the microviscosity of bilayers from natural phospholipid mixtures. AB - The thermotropic behaviour of two natural extracts of phospholipids differing by their content in saturated fatty acids and acidic polar heads were studied with the aid of the fluorescent probes : perylene and diphenylhexatriene. The main results are that the bilayers are in a fluid state in the range from -- 5 to 50 degrees C, in the presence as well as in the absence of calcium, and that misleading conclusions may be derived with the perylene probe. The effects of pH and Ca2+/H+ interaction on the microviscosity were studied. In the absence of calcium pH increase fluidizes the bilayers ; when Ca2+ is present the microviscosity is constant over a large pH range. Calculations using the Gouy Chapman theory suggest that the pH-induced microviscosity changes are controlled by the content in phosphadidylserine, diphosphatidylglycerol and phosphatidic acid of the extracts. Similarly the rigidifying effect of Ca2+ seems to be related to the content in these phospholipids. These hypothesis were confirmed by modifying the content of the extracts in these phospholipids. It is proposed that the behaviour of the two studied extracts results from the high insaturation of their fatty acids, which is typical of biological membranes. PMID- 6284256 TI - Decatenation of the kinetoplast DNA network of trypanosomes by a bacterial DNA gyrase. PMID- 6284257 TI - Comparison of different methods of recovering DNA from a methylation assay. AB - Several enzymes, for example DNA (cytosine-5-)-methyltransferase, produce relatively strong interactions with DNA and hinder the quantitative recovery of this DNA from a reaction mixture. Classical methods like the Sevag chloroform-iso amylic alcohol one or the phenol procedure lead only to a 50-60 per cent recovery of the DNA. A new procedure was worked out utilizing pancreatic RNase, proteinase K, NaOH 0.5 M treatment and trichloracetic acid precipitation which gives 85 per cent recovery of DNA. PMID- 6284258 TI - Demonstration of different complexation modes between cobalt and 5' AMP, by direct NMR observation of the low-temperature complex. AB - We report the first direct NMR observation of transition metal-nucleotide complexes. The phosphorus and proton spectra of a cobalt-5' AMP complex were observed in water, pH 7, -- 10 degrees C. This complex is different from the high temperature species : for instance its 31P chemical shift is -- 50 ppm, whereas the published value obtained indirectly for the high temperature form corresponds to ca. -- 1200 ppm. The -- 50 ppm complex is present in significant proportion at 20 degrees C and possibly at higher temperatures. Multiple complexation modes are also observed for Co-ATP. PMID- 6284259 TI - Regulation of fatty acid composition of Cl. perfringens type A cells by hydrogen and hydroxyl ions. AB - The composition of the fatty acid profiles of Cl. perfringens type A, grown on media with initial pH values from 5.5 to 9.0, was determined by the method of gas liquid chromatography. The fatty acid profiles are stabilized in 18- to 24-h cultures. Hydrogen ions stimulate the synthesis of cyclopropane carboxylic fatty acids and "desaturase" activity; hydroxyl ions inhibit these processes. The content of saturated fatty acids in the bacteria is regulated by the initial acid alkaline conditions of the medium. An increase in biomass accumulation under the influence of hydroxyl ions is coupled with a decrease in the energy supply of the bacteria. Possible mechanisms of the development of resistance to hydrogen and hydroxyl ions during the cultivation of Clostridia on a period growth medium are discussed. PMID- 6284260 TI - [Cytochromes c and P-450 as terminal acceptors in a reconstituted system of mitochondrial hydroxylation]. AB - Both cytochromes c and P-450 can be reduced in vitro by a mitochondrial hydroxylation chain consisting of flavoprotein, adrenodoxin and NADPH as electron donors. The effects of pH, ionic strength, content and stoichiometry of intermediate electron carriers on the reduction rate of cytochromes c and P-450 have been compared. The data obtained demonstrate that the reduction of these cytochromes proceeds by different ways. It has been found that cytochrome c has an inhibitory effect on cytochrome P-450 reduction. On the other hand, cytochrome P-450 remarkably activates the process of cytochrome c reduction. PMID- 6284261 TI - Alteration of interhemisphere conduction through corpus callosum in chronic schizophrenia. AB - The contralateral and ipsilateral somatosensory evoked potentials (SEP) after stimulation of the right and left median nerve were recorded in 10 patients with chronic schizophrenia and in 10 normal subjects. The ipsilateral SEP depends on the function of the corpus callosum. The latency-delay from peak 5 in the contralateral SEP to the analogous peak in the ipsilateral SEP was significantly reduced in the patients, but only from the left to the right hemisphere. This peak presumably represents interhemisphere transfer and function of corpus callosum. The amount of synaptic activity--measured by the mean amplitude of the SEP--in the right hemisphere, relative to the activity in the left hemisphere after stimulation of the right median nerve, was also greater in the patients. These differences could be interpreted as an abnormal function of the corpus callosum and as a breakdown of the lateral specialization of the left cerebral hemisphere in schizophrenia. PMID- 6284263 TI - Continuous flow-resonance Raman spectroscopy of an intermediate redox state of cytochrome C. AB - An intermediate redox state of cytochrome c at alkaline pH, generated upon rapid reduction by sodium dithionite, has been observed by resonance Raman (RR) spectroscopy in combination with the continuous flow technique. The RR spectrum of the intermediate state is reported for excitation both in the (alpha, beta) and the Soret optical absorption band. The spectra of the intermediate state are more like those of the stable reduced form than those of the stable oxidized form. For excitation of 514.5 nm, the most prominent indication of an intermediate state is the wave-number shift of one RR band from 1,562 cm-1 in the stable oxidized state through 1,535 cm-1 in the intermediate state to 1,544 cm-1 in the stable reduced state. For excitation at 413.1 nm, a band, present at 1,542 cm-1 in the stable reduced state but not present in the stable oxidized state, is absent in the intermediate state. We interpret the intermediate species as the state where the heme iron is reduced but the protein remains in the conformation of the oxidized state, with methionine-80 displaced as sixth ligand to the heme iron, before relaxing to the conformation of the stable reduced state, with methionine-80 returned as sixth ligand. PMID- 6284262 TI - Phosphoprotein phosphatase activity of sea urchin spermatozoa. PMID- 6284264 TI - Influence of cellular and paracellular conductance patterns on epithelial transport and metabolism. AB - Theoretical analysis of transepithelial active Na transport is often based on equivalent electrical circuits comprising discrete parallel active and passive pathways. Recent findings show, however, that Na+ pumps are distributed over the entire basal lateral surface of epithelial cells. This suggests that Na+ that has been actively transported into paracellular channels may to some extent return to the apical (mucosal) bathing solution, depending on the relative conductances of the pathways via the tight junctions and the lateral intercellular spaces. Such circulation, as well as the relative conductance of cellular and paracellular pathways, may have an important influence on the relationships between parameters of transcellular and transepithelial active transport and metabolism. These relationships were examined by equivalent circuit analysis of active Na transport, Na conductance, the electromotive force of Na transport, the "stoichiometry" of transport, and the degree of coupling of transport to metabolism. Although the model is too crude to permit precise quantification, important qualitative differences are predicted between "loose" and "tight" epithelia in the absence and presence of circulation. In contrast, there is no effect on the free energy of metabolic reaction estimated from a linear thermodynamic formalism. Also of interest are implications concerning the experimental evaluation of passive paracellular conductance following abolition of active transport, and the use of the cellular voltage-divider ratio to estimate the relative conductances of apical and basal lateral plasma membranes. PMID- 6284265 TI - Dielectric constant dependence of biological oxidation-reduction. 1. A model of polarity-dependent ferrocytochrome c oxidation. AB - A theoretical model for the effect of the dielectric constant (c) of the solvent medium on ferrocytochrome c oxidation by ferricyanide is developed to account for the observed variations of the rate constant (k) of reactions in aqueous binary mixtures with alcohols (less than 5-10 mol% ethanol and propranolol). A correlation between k and c is found if ln k is expressed as a function of the Kirkwood parameter (c-1)(2c+1). The results of calculations indicate that the use of the 'overall dipole moment' of cytochrome c in oxidoreduction studies is likely to be unreliable. Instead, the decrease in k in alcohol/water mixtures is best explained--in conformity with Onsager's theory of the reaction field--by a polarity effect on the dipole moment of the cytochrome c heme upon diffusion of the polar solvent molecules into the low dielectric constant heme crevice. PMID- 6284267 TI - Microscopic calculation of ion-transport rates in membrane channels. AB - A method, based on rate theory, is described by which transport rates in ion channels can be calculated using only microscopic parameters, such as atomic coordinates, force constants and intermolecular energy parameters. The channel is treated as a system of elastically bound ligands interacting with the ion by coulombic and Lennard-Jones forces. Jump frequencies of the ion are obtained from the potential mean force which represents a thermal average over the different configurations of the ligand system. The method is illustrated by application to a special channel model, helical arrangement of dipolar ligands, which can be tilted toward the channel axis against harmonic restoring force. The jump frequency is found to be a non-monotonous function of ion radius. Furthermore, the ion specificity of the channel strongly depends on whether the ligand system is 'hard' or 'soft', i.e., on the extent to which the interaction with the ion can lead to a reorientation of the ligand groups. PMID- 6284266 TI - Electric properties and structure of DNA-restriction fragments from measurements of the electric dichroism. AB - The electric dichroism of 17 homogeneous DNA fragments, ranging in size from 43 to 4362 base-pairs, has been analyzed in high electric fields. The orientation of the small fragments can be described in terms of an induced dipole moment, whereas the large fragments are oriented according to a constant dipole mechanism. In the intermediate size range, DNA orients according to an induced dipole mechanism at low field strengths and according to a constant dipole mechanism at high field strengths. From these observations we propose an orientation mechanism with a saturating induced dipole. The induced dipole observed at low field strengths is saturated at a field strength Eo within a transition range Em to give a constant dipole moment at high field strengths. These parameters together with the polarizability and the limit reduced dichroism are evaluated by a least-squares analysis of the experimental data. Eo and Em are found to decrease with increasing chain length from Eo approximately 40 kV/cm (Em approximately 14 kV/cm) at 65 base-pairs to 10 kV/cm (6 kV/cm) at 194 base-pairs. The polarizability is found to increase with the square of the chain length, whereas the saturated dipole increases with chain length N at low N and goes to a limit value at high N. The temperature dependence of the orientation parameters is found to be very small. The values obtained for the limit dichroism are between -1.0 and -1.3 for chain lengths between 60 and 1000 base-pairs, whereas values around -1.4 are observed at chain lengths greater than 1000 base-pairs. These data indicate that electric fields extend the contour of DNA strands at high chain lengths from a weakly bent to a more linear form. The variations of the limit dichroism observed for short fragments suggest sequence-dependent differences in the secondary structure of the helix. The experimental results are compared with numerical calculations based on simple polyelectrolyte models. For short fragments the magnitude of several electrochemical parameters can be adequately explained by a polarization of the ion cloud around the DNA molecules. However, these polyelectrolyte models do not adequately describe the observed chain length dependence of the orientation phenomena. PMID- 6284269 TI - Effect of alkylation on the secondary structure of DNA. AB - S1 nuclease hydrolysis and bezoylated naphthoylated DEAE-cellulose (BND cellulose) chromatography have been used to demonstrate that alkylation of DNA by dimethyl sulfate at neutral pH leads to the production of partially denatured molecules under conditions where no significant depurination occurs. DNA was alkylated with increasing concentrations of the alkylating agent, and subjected to enzymatic degradation and binding to BND cellulose. An increasing degree of DNA hydrolysis and adherence to BND cellulose was seen. On hydroxyapatite chromatography the alkylated DNA still eluted at the position of double-stranded molecules suggesting the presence of partially denatured regions. The presence of salt had a preventive effect on such denaturation. PMID- 6284268 TI - Bohr-effect and pH-dependence of electron spin resonance spectra of a cobalt substituted monomeric insect haemoglobin. AB - The monomeric haemoglobin IV from Chironomus thummi thummi (CTT IV) exhibits an alkaline Bohr-effect and therefore it is an allosteric protein. By substitution of the haem iron for cobalt the O2 half-saturation pressure, measured at 25 degrees C, increases 250-fold. The Bohr-effect is not affected by the replacement of the central atom. The parameters of the Bohr-effect of cobalt CTT IV for 25 degrees C are: inflection point of the Bohr-effect curve at pH 7.1, number of Bohr protons -- deltalog p1/2 (O2)/deltapH = 0.36 mol H+/mol O2 and amplitude of the Bohr-effect curve deltalogp1/2 (O2) = 0.84. The substitution of protoporphyrin for mesoporphyrin causes a 10 nm blue-shift of the visible absorption maxima in both, the native and the cobalt-substituted forms of CTT IV. Furthermore, the replacement of vinyl groups by ethyl groups at position 2 and 4 of the porphyrin system leads to an increase of O2 affinities at 25 degrees C which follows the order: proto less than meso less than deutero for iron and cobalt CTT IV, respectively. Again, the Bohr-effect is not affected by the replacement of protoporphyrin for mesoporphyrin or deuteroporphyrin. The electron spin resonance (ESR) spectra of both, deoxy cobalt proto- and deoxy cobalt meso CTT IV, are independent of pH. The stronger electron-withdrawing effect by protoporphyrin is reflected by the decrease of the cobalt hyperfine constants coinciding with gparallel = 2.035 and by the low-field shift of gparallel. The ESR spectra of oxy cobalt proto- and oxy cobalt meso-CTT IV are dependent of pH. The cobalt hyperfine constants coinciding with gparallel - 2.078 increase during transition from low to high pH. The pH-induced ESR spectral changes correlate with the alkaline Bohr-effect. Therefore, the two O2 affinity states can be assigned to the low-pH and high-pH ESR spectral species. The low-pH form (low affinity state) is characterized by a smaller, the high-pH form (high-affinity state) by a larger cobalt hyperfine constant in gparallel. The correlation of the cobalt hyperfine constants of the oxy forms with the O2 affinities is discussed for several monomeric haemoglobins. The Co-O-O bond angle in cobalt oxy CTT IV is characterized by an ozonoid type of binding geometry and varies little during the pH-induced conformation transition. Due to the lack of the distal histidine in CTT IV no additional interaction via hydrogen-bonding with dioxygen is possible; this is reflected by the cobalt hyperfine constants. PMID- 6284270 TI - Adrenaline reactivation of muscle phosphorylase after deactivation during phasic contractile activity. AB - The increase in % phosphorylase a associated with muscle contraction is reversed after a few minutes despite continued contractile activity. The present study was undertaken to determine whether adrenaline can activate phosphorylase after deactivation has occurred during prolonged stimulation of contraction. We found that adrenaline reactivates phosphorylase and glycogenolysis in rat fast-twitch and slow-twitch muscles that have been stimulated to contract for 15 min. The reactivation of glycogenolysis by adrenaline could have importance in fight-or flight situations that call for an increase in exercise intensity. PMID- 6284271 TI - Effect of calcium ion on the mechanical properties of lipid bilayer membrane. AB - Elastic properties of dipalmitoyl phosphatidylcholine bilayers were studied by an ultrasonic technique as a function of the concentration of calcium ion. The ultrasonic velocity showed a broad minimum at about 10 mM CaCl2, indicating that the bulk modulus of membrane is smallest at this calcium ion concentration. Since the amount of calcium ions adsorbed to the membrane monotonously increases, calcium ions have two kinds of effects on the phosphatidylcholine membrane, namely softening effect at lower concentration and hardening at higher concentration. It is suggested that the softening is due to some structural change of hydrated water and the hardening is due to chelation of lipid molecules by adsorbed calcium ions. The estimation of bulk modulus and Young's modulus revealed that Young's modulus and rigidity are much smaller than the bulk modulus even in the gel phase. PMID- 6284272 TI - [Reaction of peripheral blood leukocytes to wound processes during phytohemagglutinin therapy in rabbits with different hypersensitivity of delayed type]. AB - Altogether 165 rabbits were distributed into strongly and poorly reacting groups in terms of delayed type hypersensitivity (DTH) to phytohemagglutinin (PHA). It was established that leukocyte response to wound process correlated with the intensity of DTH. The strongly reacting animals manifested of DTH. The strongly reacting animals manifested an adequate response (leukocytosis, neutrophilosis, nomocytosis, lymphocytosis, a decrease in myeloperoxidase and enhancement of phagocytic activity of neutrophils), while the poorly responding animals an in adequate one (passivity of the leukocytic parameters with the exception of monocytosis). Application of the liniment containing PHA (3.3 micrograms per g base, Difco, USA) does not change the response in the strongly reacting animals and essentially corrects all the leukocytic parameters in the poorly reacting animals with the exception of phagocytic activity of neutrophils. From the leukocytic response pattern a conclusion is made about multiple defect of the leukocytic system under poor DTH to PHA and about the leading role of monocytes in correcting the defect by PHA. PMID- 6284273 TI - [Juxtaglomerular apparatus and interstitial cells of the kidney medulla after the administration of certain drugs and use of hyperbaric oxygenation]. AB - Electron microscopy was used to examine the status of the juxtaglomerular apparatus (JGA) and interstitial cells (IC) 3 and 24 hours after administration of furosemide (10 mg/kg), indomethacin (10 mg/kg), venoruton (500 mg/kg) and trental (100 mg/kg), and after 1,6 an 12 sessions of hyperbaric oxygenation. To evaluate objectively the results of examining the JGA, use was made of a method devised by the authors of a mathematical appraisal of granulation from the density of the epithelioid cells. Granulation of 50 IC from each animal was calculated on semi-thin araldite sections stained methylene blue-azur II-fuchsin. The results indicate that all the types of exposure including hyperbaric oxygenation produced JGA activation whose degree varied depending on the time elapsed after exposure. An apparently great increase in the JGA activity was detected after injection of furosemide and indomethacin. All the drugs with the exception of furosemide entailed granule accumulation after 3 hours, followed by the recovery of their amount after 24 hours. Furosemide injection produced a reverse effect. PMID- 6284274 TI - [Effect of S-adenosine homocysteine and dibutyril-3',5-AMP on lysosomal membranes of eye tissue in ocular herpes]. PMID- 6284275 TI - [Effect of antidepressants with different chemical structures on the uptake and release of norepinephrine in sections of the rat cerebral cortex]. AB - Experiments with slices of the rat cortex were made to study the interaction between the ability of the antidepressants to inhibit the reverse uptake of 14C noradrenaline and to inhibit its presynaptic release. The antidepressants studied are distributed into 3 groups according to the ratio of effective concentrations that block the uptake and enhance the release of 14C-noradrenaline from the slices. The first group includes the antidepressants (melipramine, chlorimipramine, Lu 5-003, Lu 3-010, S-394, pyrazidol) that have a ratio close to I and in whose mechanism of aminopotentiating action the main component is unlikely to be distinguished. The second group with a ratio less than I is represented by the substances (nortryptyline, desipramine) whose mechanism of the aminopotentiating action is determined by the inhibitory effect on the neurotransmitter reverse uptake. As to the 3d group antidepressants (thyroliberin, iprindol, noveril, C-356, C-395, amitryptyline), of great importance is their effect on the presynaptic neurotransmitter release from the terminals of the axons of noradrenergic neurons. PMID- 6284276 TI - [Effect of creatine phosphate on the duration of action potentials and extent of myocardial contraction of hypoxic frog heart]. PMID- 6284277 TI - [Role of cyclic nucleotides in the mechanism of action of enkephalins]. PMID- 6284278 TI - [Experimental study of the effect of nonachlazine on the metabolism of the ischemic myocardium]. PMID- 6284279 TI - [Postural asymmetry induced by enkephalin analogs]. PMID- 6284280 TI - [Sensitivity of retinal pigments to the melanotropic effect of ACTH in normal and mutant rats with hereditary retinal dystrophy]. PMID- 6284281 TI - [Role of cAMP in regulating the shape of thrombocytes]. AB - Preincubation of platelet suspension with dbcAMP during 2 minutes inhibited the ADP-induced change in platelet shape. An increase of the cAMP content in the cell induced by papaverine inhibiting phosphodiesterase also blocked the change in the shape caused by ADP and prostaglandin E2 (PGE2). Incubation of platelets in medium with pH 9.0 or addition of PGE2 produced a stable change in the shape of the cells. The action of dbcAMP or papaverine on the changed cells accelerated the recovery of the native shape and the content of membrane-bound calcium. PMID- 6284282 TI - [Effect of the "memory peptide" ACTH 5-10 on the self-stimulation reaction of rabbits]. AB - Experiments on rabbits were made to study variation in the frequency of the self stimulation reaction from the lateral hypothalamus under the effect of the corticotropin fragment ACTH5-10. Intraperitoneal administration of the peptide in a dose of 50 micrograms/kg that causes the improved training in different behavioral models produces no significant effect on the mechanism of intracranial positive reinforcement. Intraventricular injection of 5 microliters of 0.9% NaCl leads to a short-term suppression of the self-stimulation reaction. Administration of 50 pcM/kg ACTH5-10 in the same volume of physiological saline completely abolishes the inhibitory action of the intraventricular injection itself. PMID- 6284283 TI - [Effect of the GABA receptor blockader bicuculline on the action of fenibut and diazepam]. AB - Fenibut (100 mg/kg) suppressed the motility and emotional reactivity but did not possess any antiaggressive properties, whereas diazepam (2.5 mg/kg) did increase the motility and reduced aggressiveness in rats. Bicuculline (1.25 mg/kg) itself did not cause any behavioral changes, although it was capable of antagonizing the effects of diazepam and to potentiate those of fenibut. Bicuculline attenuated the GABA rise but did not influence the increased content of HVA and DOPAC after fenibut. The importance of the bicuculline-insensitive mechanisms in the action of fenibut and bicuculline-sensitive ones in the action of diazepam is discussed. PMID- 6284284 TI - New approaches to the detection of myeloperoxidase deficiency. AB - Family studies on myeloperoxidase (MPO) deficiency have been carried out by quantitating the peroxidase activity of granulocyte preparations with three methods, namely guaiacol peroxidation, alanine decarboxylation, and spectroscopic analysis. The guaiacol assay failed to show a definite pattern of inheritance in two families with MPO-deficient subjects. Surprisingly, the granulocytes of three histochemically MPO-negative subjects had a peroxidase activity either half or even higher than that of control subjects. The peroxidase activity of these granulocyte preparations in these three subjects showed a positive correlation to the number of eosinophils. The possibility then considered was that eosinophils may have obscured the true pattern of inheritance in this assay. Two other methods of MPO assay, which are not influenced by the presence of eosinophil peroxidase (EPO), were therefore devised. One is based on the ability of MPO, but not EPO, to catalyze decarboxylation of L-alanine in the presence of Triton X 100, and the other relies on the different spectral properties of the two peroxidases. The results obtained with these two methods (1) were strictly comparable, (2) allowed detection of both totally and partially MPO-deficient subjects, (3) differed profoundly from those obtained with the guaiacol method when eosinophil-containing granulocyte preparations were used, and (4) revealed a pattern of autosomal recessive inheritance in the two families studied. The results of the three methods were comparable when eosinophil-free granulocyte preparations were assayed. It is concluded that failure to show a pattern of inheritance in some instances of primary MPO deficiency, or deviations from the autosomal recessive mode of transmission of this defect, may be attributed to interference by EPO. It is proposed that peroxidase assay methods not subject to EPO interference, such as the two described in this article, may be used, particularly in the detection of heterozygote subjects for MPO deficiency in the presence of high eosinophil counts. PMID- 6284285 TI - Studies on the mechanism of the inhibition of platelet aggregation and release induced by high levels of arachidonate. AB - We have previously reported that arachidonic acid induced a biphasic pattern of platelet aggregation and the release of both dense and alpha-granule components. Low levels of arachidonate (0.025--0.1 mM) specifically induced aggregation and release, while high concentrations (0.15--0.35 mM) caused a progressive inhibition of these platelet responses in human gel-filtered platelets (GFP). We now report studies of the mechanism(s) responsible for this arachidonate-induced turn-off of platelet function. Electron micrographic studies demonstrated that there was no gross damage to the platelets during the turn-off. Active synthesis of malondialdehyde and thromboxane A2 was seen at the high arachidonate levels, despite the inhibition of aggregation. Furthermore, GFP inhibited by 0.25 mM arachidonate were capable of undergoing aggregation and serotonin release in response to other stimuli, such as collagen or thrombin. Thus, GFP appeared to be metabolically intact and functional during the inhibiton by high arachidonate levels. Thin-layer chromatographic studies revealed that prostaglandin metabolism was not changed at the high arachidonate levels. In addition, indomethacin (20 microM) did not abolish the arachidonate-induced inhibition of platelet function. Therefore, the inhibitory effect of high arachidonate did not depend on its conversion to other prostaglandin products. Platelet cyclic AMP levels increased twofold at the high arachidonate concentrations (1.3 +/- 0.3 pmole/10(8) platelets at peak aggregation, compared with 2.9 +/- 0.4 pmole/10(8) platelets at inhibition by 0.25 mM arachidonate, p less than 0.001). Prostaglandin-D2, a platelet inhibitor known to increase cyclic AMP, generated a similar rise (to 2.4 +/- 0.2 pmole/10(8) platelets). Thus, the magnitude of the arachidonate-induced increase in platelet cyclic AMP levels can account for the inhibition of aggregation and release. PMID- 6284286 TI - The significance of ecto-5'-nucleotidase in lymphoid blast crisis of chronic myeloid leukemia. PMID- 6284287 TI - Elevation of brain cyclic nucleotides during acute dieldrin exposure. PMID- 6284288 TI - The association of hepatitis B surface antigen (HBs Ag) with hepatocellular carcinoma in Espirito Santo State, Brazil. AB - 1. The occurrence of hepatitis B surface antigen (HBs Ag) was investigated in 46 cases of hepatocellular carcinoma (HCC) diagnosed in Vitoria, Espirito Santo State, Brazil. 2. Twenty cases were patients in which the HCC was diagnosed by biopsy, laparoscopy or on autopsy from 1977 to 1980. HBs Ag was measured in the serum by reverse passive hemagglutination (13 cases) and radioimmunoassay (7 cases). Twelve cases (60%) were HBs Ag positive. 3. The presence of HBs Ag was studied in 26 cases diagnosed on autopsy using paraffin-embedded liver sections stained with orcein. Eleven cases (42%) showed orcein granules in hepatocytes surrounding the tumor. In only one case were orcein-positive granules detected in the tumor cells. 4. The 50% incidence of HBs Ag among patients with HCC is higher than the occurrence of HBs Ag in the general population of the State of Espirito Santo, i.e. 3.5% among 1300 blood donors to the University Hospital. 5. These data provide additional evidence for the hypothesis that there is a relationship between the hepatitis B virus and the etiology of HCC. PMID- 6284289 TI - The effects of prazosin, phentolamine and phenoxybenzamine on inhibitory alpha adrenoceptors in the guniea-pig isolated ileum. AB - 1 The relaxant effect of noradrenaline on strips of guinea-pig isolated terminal ileum was blocked by pretreatment with prazosin, phentolamine, yohimbine and phenoxybenzamine. 2 The presence of a very high concentration of noradrenaline (50 micrometers) during exposure to the blocking agent protected against the blocking effect of the drugs. 3 Yohimbine, prazosin and phentolamine partially protected against irreversible blockade by phenoxybenzamine. 4 Spontaneous release of acetylcholine in the unstimulated ileum was blocked by noradrenaline (0.05-5.9 micrometers) this effect of noradrenaline was antagonized by phentolamine (0.13-26 micrometers) and yohimbine (0.051-0.51 micrometers) but not prazosin (0.53-5.3 micrometers) or phenoxybenzamine (4.2-42 nm). All four antagonists reversed the noradrenaline-induced relaxation of the ileum. 5 Acetylcholine output in the transmurally stimulated ileum was inhibited by noradrenaline. This effect of noradrenaline was antagonized by phentolamine and yohimbine but not by prazosin or phenoxybenzamine. The first two antagonists blocked the noradrenaline-induced inhibition of evoked twitches of the ileum while the last two had no effect. 6 The results show (a) that prazosin has no effect on presynaptic alpha-adrenoceptor located on cholinergic nerve endings in the guinea-pig ileum and (b) that prazosin, phentolamine and phenoxybenzamine act on the same subgroup of postsynaptic alpha-adrenoceptors on the smooth muscle of the guniea-pig ileum. PMID- 6284290 TI - In vitro studies on an antagonist of parathyroid hormone [Nle-8, Nle-18, Tyr 34]bPTH-(3-34)amide. AB - 1 The actions of parathyroid hormone (PTH) are antagonized in vitro by the peptide [Nle-8, Nle-18, Tyr-34]-bPTH-(3-34)amide, an analogue of PTH. In this paper, the actions of the inhibitory peptide were investigated in vivo. 2 Native parathyroid hormone (bPTH-(1-84)), administered i.v. (0.17-1.51 nmol in volume of 0.3 ml) to 7 day old chicks produced hypercalcaemia but administration of the analogue in doses up to 173 nmol was ineffective in this respect. 3 The analogue failed to antagonize the hypercalcaemia produced by bPTH-(1-34) when injected, in 10 fold molar excess, 2 min before or simultaneously with bPTH-(1-34). 4 Normocalcaemia was restored in parathyroidectomized rats by intravenous infusion of bPTH-(1-84) at 32 pmol kg-1 h-1. Addition of the analogue to the infusion fluid in a 200 fold molar excess did not affect the concentrations of calcium and phosphate in the plasma, cyclic adenosine 3',5'-monophosphate (cyclic AMP) in the urine or phosphate clearance but produced a significant (P less than 0.05) rise in urinary calcium clearance. 5 The results suggest that the peptide [Nle-8, Nle 18, Tyr-34]-bPTH-(3-34)amide does not antagonize the actions of PTH in vivo and demonstrate an important dichotomy between in vitro and in vivo biological properties of the PTH analogue. PMID- 6284291 TI - Regional rat brain benzodiazepine receptor number and gamma-aminobutyric acid concentration following a convulsion. AB - 1 Following administration to rats of an electroconvulsive shock (ECS) which resulted in a major tonic-clonic seizure, no changes in [3H]-diazepam binding characteristics were observed in cortex or hippocampus with either a well washed membrane preparation or a crude synaptosomal preparation. 2 No changes were observed in [3H]-diazepam binding in any other brain region examined 30 min after an ECS. 3 Thirty min following an ECS, regional brain gamma-aminobutyric acid (GABA) concentrations increased in hippocampus, cortex and hypothalamus. Only in the hippocampus did the increase occur within 5 min of the seizure. 4 Similar increases in GABA concentration were seen after a bicuculline-induced seizure but not after seizure induced by flurothyl; both treatments produced a tonic-clonic seizure. 5 Pretreatment of the rats with (+)-propranolol 5 min before the ECS abolished the tonic extension and prevented the brain GABA concentration changes that occur 30 min after the seizure. 6 No increase in GABA concentration was seen in hippocampus, cortex and hypothalamus 30 min after the final ECS of a course of 10 ECS given once daily for 10 days. In contrast a marked increase in striatal GABA concentration was observed. 7 These changes in GABA biochemistry following a seizure are discussed in relation to the post-ictal rise in seizure threshold that is occurring at the same time. PMID- 6284292 TI - Desensitization by terbutaline of beta-adrenoceptors in the guinea-pig soleus muscle: biochemical alterations associated with functional changes. AB - 1 The effects of adrenaline and terbutaline on cyclic adenosine 3',5' monophosphate (cyclic AMP) content, 22Na-efflux, 42K-influx and subtetanic contractions have been assessed in soleus muscles isolated from guniea-pigs which had been maintained on food with or without terbutaline for 5 days. 2 Terbutaline and adrenaline increased cyclic AMP content and suppressed subtetanic contractions, and regression analysis indicates a statistically significant correlation between these two effects (P less than 0.01). 3 In muscles obtained from terbutaline-treated animals, the effects of terbutaline and adrenaline on cyclic AMP content, active Na-K-transport and subtetanic contractions were all considerably suppressed, but insulin stimulated 22Na-efflux and affected subtetanic contractions to the same extent as in the muscles obtained from the control group. 4 The results suggest that terbutaline treatment leads to a reduction in the number of beta 2-adrenoceptors in skeletal muscle or an impairment of their function. 5 The results provide further support for the idea that the effect of adrenaline or insulin on skeletal muscle contractions is the outcome of stimulation of active Na-K-transport. PMID- 6284293 TI - An examination of the pre- and postsynaptic alpha-adrenoceptors involved in neuroeffector transmission in rabbit aorta and portal vein. AB - 1 The alpha-adrenoceptor agonists, clonidine and xylazine, reduced and the alpha antagonists, yohimbine an rauwolscine, increased the stimulation-evoked tritium overflow from rabbit aorta and portal vein pre-incubated with [3H]-noradrenaline. 2 Based on an order of agonist potency of clonidine greater than xylazine greater than phenylephrine and antagonist potency of rauwolscine = yohimbine greater than prazosin, the presynaptic receptor mediating these effects is of the alpha 2 type. 3 In the aorta, stimulation-evoked contractions were abolished by prazosin (0.1 micrometers) and potentiated by rauwolscine and yohimbine in concentrations that increased the stimulation-evoked overflow tritium. 4 In the portal vein, prazosin was less potent in reducing, and rauwolscine and yohimbine failed to potentiate, the stimulation-evoked contraction. 5 In experiments in which tissues were pre-exposed to phenoxybenzamine (30 nM) to block some of the postsynaptic alpha-receptors, rauwolscine in concentrations that increased stimulation-evoked tritium overflow, reduced the evoked contraction in the portal vein but not in the aorta. 6 It is concluded that presynaptic alpha 2-autoreceptors are present in both tissues and that the postsynaptic alpha-receptors which mediate nerve stimulation-evoked contractions are alpha 1 in the aorta but a mixture of alpha 1 and alpha 2 in the portal vein. PMID- 6284294 TI - High concentrations of thyroid-stimulating hormone in untreated glucocorticoid deficiency: indication of primary hypothyroidism? PMID- 6284296 TI - Efficacy of pertussis vaccination in England. Report from the PHLS Epidemiological Research Laboratory and 21 area health authorities. AB - A national assessment of the efficacy of pertussis vaccination was made in 21 area health authorities in England. Notification rates for children given either three doses of diphtheria/tetanus/pertussis vaccine or diphtheria/tetanus vaccine were studied during the two and a half-year period January 1978-June 1980. A survey of home circumstances showed that the two vaccine groups could be validly compared. Home visits were made to assess the severity of the illness, the attack rates in home contacts, and to take pernasal swabs. Pertussis vaccination was found to be of outstanding value in preventing serious disease. Nevertheless, its effect in controlling whooping cough is limited by the fact that protection for home contacts is less satisfactory, and by the occurrence of mild cases in vaccinated children which may contribute to the spread of the disease. PMID- 6284297 TI - ABC of diabetes: diabetic emergencies. PMID- 6284295 TI - Maternal anti-D concentrations and outcome in rhesus haemolytic disease of the newborn. AB - The relation between maternal anti-D concentrations, measured against the British working standard, and outcome of rhesus-sensitised pregnancies was studied. There is a clear relation between increasing anti-D concentrations and the chance of a severely affected baby. Of those pregnancies (78) where serial anti-D concentrations remained below 4 IU/ml, no baby had a cord haemoglobin below 10 g/dl and three had exchange transfusions. In contrast, of those mothers (106) with anti-D concentrations above 4 IU/ml, 23 had babies with a cord haemoglobin below 10 g/dl and 79 babies had exchange transfusions. It is suggested that those pregnancies where anti-D concentrations remain below 4 IU/ml represent a relatively safe group in which amniocentesis may be avoided. PMID- 6284298 TI - Antibody titres to African horse sickness, swine vesicular disease and foot-and mouth disease viruses in samples from Equidae in Malta, 1975. PMID- 6284299 TI - Blockade of transmission in rat sympathetic ganglia by a toxin which co-purifies with alpha-bungarotoxin. AB - Bungarus multicinctus venom was fractionated into its toxin components using ion exchange chromatography on CM-Sephadex. According to previous reports, rechromatography of fraction II on a CM cellulose column yields chemically homogenous alpha-bungarotoxin (II2) of molecular weight 9000. However, in our hands, using the identical purification procedure, two discrete proteins of molecular weight 9000 and 15,000 were obtained as demonstrated by SDS gel electrophoresis. Subsequent fractionation of this alpha-bungarotoxin fraction (II2) was achieved on Sephadex G-50. The 9000 weight component (labelled II-S2) was identical to alpha-bungarotoxin; at a concentration of 1 microgram/ml it blocked transmission at the neuromuscular junction but did not block nicotinic responses in rat sympathetic ganglia. Very different properties were exhibited by II-SI, the 15,000 molecular weight component; it inhibited ganglionic transmission but was ineffective at the neuromuscular junction at the same concentration (1 microgram/ml). BGT II-S1 was equipotent in blocking the ganglionic action potential in the presence or absence of eserine; thus, it is not acting as an acetylcholinesterase by increasing acetylcholine breakdown. In the presence of toxin, [3H]choline incorporation into ganglionic acetylcholine during preganglionic stimulation was not altered, suggesting that the toxin did not block transmission by a presynaptic mechanism. Thus, the site of action of the toxin appears to be postsynaptic although it did not affect depolarization of the ganglia induced by carbachol. PMID- 6284302 TI - Inhibition by morphine and beta-endorphin of vasopressin release evoked by electrical stimulation of the rat medial basal hypothalamus in vitro. PMID- 6284301 TI - Two modes of activity of nicotinic acetylcholine receptor channels in sympathetic neurons. AB - Spectral analysis of acetylcholine (ACh) noise was performed in voltage-clamped neurons of the isolated rabbit superior cervical ganglion at 34-37 degrees C and at membrane potential--80 mV. Two modes of activity were found in the ionic channels of nicotinic ACh receptors, with mean channel life-times of for fast operating channels and for slow-operating channels. Excitatory postsynaptic current (EPSC) decays exponentially with a time constant which is very close to , indicating that the slow-operating channel activity determines the duration of EPSC. The mean value of conductance of single nicotinic ACh-receptor channel is 36 more or less 3 pS. PMID- 6284300 TI - Intracellular recordings from serotonergic dorsal raphe neurons: pacemaker potentials and the effect of LSD. AB - Intracellular recordings in vivo from serotonergic dorsal raphe neurons of the rat brain reveal that these cells undergo a pronounced postspike hyperpolarization followed by a gradual interspike depolarization leading to the succeeding spike. Such repetitive cycles of interspike hyperpolarization and depolarization, which can be termed "pacemaker potentials', can account for the automaticity of these cells. When serotonergic neuronal firing is inhibited by LSD, such pacemaker potentials no longer occur and the cells remain in an hyperpolarized state. PMID- 6284303 TI - Attenuation by chronic imipramine treatment of [3H]clonidine binding to cortical membranes and of clonidine-induced hypothermia: the influence of central chemosympathectomy. AB - Central chemosympathectomy with intraventricular injection of 250 microgram of 6 hydroxydopamine, resulting in depression of cerebral noradrenaline level by 75% led to an increase in specific binding of [3H]clonidine to cerebral cortical membranes, but did not affect the hypothermic response to clonidine. Chronic imipramine treatment depressed [3H]clonidine binding and attenuated clonidine induced hypothermia similarly in non-chemosympathectomized and chemosympathectomized rats. PMID- 6284304 TI - The effect of Mg2+ and Ca2+ on the excitability of Pacinian corpuscles. AB - The effect of altering extracellular Ca2+ and Mg2+ levels on electrical properties of Pacinian corpuscles of adult cat has been examined. High Mg2+ solution (5--15 mM) increased the excitability of the receptors and the amplitude of the receptor potential. Ca2+-free EDTA solution caused an increase in the amplitude of the receptor potential in the first 60 min and then its decrease. Mg2+ ions slow down or prevent the depressant effect of the Ca2+ lack on the receptor potential. The phase of the receptor potential amplitude increase in Ca2+-free media was reversibly blocked by tetraethylammonium ions. Treatment with 2,4-dinitrophenol produced a transient increase in the receptor potential amplitude. The threshold of the mechanosensitive receptor membrane was unaffected in Ca2+-free or Mg2+-rich solution. The results suggest that divalent cations are not directly involved in the process of mechanoelectric transformation in Pacinian corpuscles. PMID- 6284305 TI - Response of nucleus accumbens neurons to amygdala stimulation and its modification by dopamine. AB - Extracellular single unit recordings were obtained from the nucleus accumbens of urethane anesthetized rats. It was found that electrical stimulation of the basal lateral and basal medial nuclei of the amygdala produced strong excitatory responses in neurons of the nucleus accumbens, in particular the medial region. Latencies of activation were relatively short with a mean of 10.7 ms. Dopamine applied iontophoretically had a marked attenuating effect on the excitatory response of nucleus accumbens neurons to amygdala stimulation. The spontaneous activity of all neurons recorded from the nucleus accumbens was also suppressed by dopamine, but the excitatory response was more sensitive to dopamine inhibition than the spontaneous activity. Neurons in the nucleus accumbens showed a variety of responses to single-pulse electrical stimulation of the ventral tegmental area (VTA). Some units in the nucleus accumbens received convergent inputs from both the amygdala and the VTA. Stimulation of the VTA also attenuated the response of nucleus accumbens neurons to excitatory inputs from the amygdala. A train of 10 pulses (0.15 ms, 200--600 microA) at 10 Hz delivered to the VTA at 100 ms before stimulation of the amygdala caused attenuation of the original excitatory response. The attenuating effect could be observed irrespective of whether individual single-pulse stimulation of the VTA elicited a response in that particular accumbens neuron or not. 6-Hydroxydopamine injected into the VTA 2 days prior to the recording experiment, or haloperidol injected intraperitoneally 1 h before the recording session, abolished this attenuating effect. However, responses to single-pulse stimulations of the VTA were not abolished. The results suggest that the attenuation of the excitatory response to amygdala stimulation was due to the release of dopamine from mesolimbic dopaminergic neurons. Responses to single-pulse stimulations of the VTA were probably due to activation of non-dopaminergic neurons projecting from the same area. It is suggested as a working hypothesis that this inhibitory effect of dopamine may be an important function of the mesolimbic dopamine pathway in modulating the extent to which limbic structures can exert an influence on the motor system through the accumbens. PMID- 6284306 TI - Attenuation of evoked field potentials from dentate granule cells by low glucose, pyruvate + malate, and sodium fluoride. AB - Extracellular field potentials were evoked from granule cells of the dentate gyrus by stimulation of the perforant path of the hippocampal slice, superfused with medium containing 10 mM glucose. Decreasing the glucose concentration to 2 mM caused a large but reversible attenuation of the rate of rise of population excitatory postsynaptic potential (EPSP) and of the population spike amplitude. Similar behaviour was observed in the presence of 5 mM glucose, although the decrease in EPSP was much less marked. Substitution of pyruvate + malate (2 mM, 0.2 mM) for glucose in the superfusion medium strongly depressed both EPSP and population spike. Sodium fluoride (1 mM) attenuated the population spike without affecting the EPSP. It is suggested that the behaviour of the evoked granule cell responses in the presence of lowered glucose or alternative substrates does not correlate with the energy state of the tissue. PMID- 6284307 TI - CNS control of active sodium transport in muscle during progressive hypokalemia in the rat. AB - The degree of plasma hypokalemia was graded with the duration of a potassium deficient diet. The intracellular Na+ and K+ contents ([Na]i and [K]i) of the innervated soleus muscle of hypokalemic rats were determined and plotted as a function of plasma K+ concentration following a potassium deficient diet during 1 8 weeks. The progression of plasma hypokalemia was highly correlated with both [Na]i accumulation and [K]i loss in the muscle. Denervation of muscles in the hypokalemic rat resulted in a rapid activation of the Na-pump in the denervated soleus muscle regardless of the degree of plasma hypokalemia. This pump activation in the denervated muscle was chronically maintained throughout the hypokalemia. Restoration of a normal, K+ containing, diet for 4 days in hypokalemic rats resulted in a complete recovery of normal Na+ and K+ contents in both the innervated muscle and plasma, while the [Na]i and [K]i in the denervated muscle was unaffected. The relationship between the CNS-induced inhibition of the muscle Na-pump and the plasma K+ levels in hypokalemic rats is discussed. PMID- 6284308 TI - An electrophysiological study of early retinotectal projection patterns during optic nerve regeneration in Hyla moorei. AB - The sequence of regeneration of the optic nerve into the tectum has been studied using electrophysiological visual mapping in the frog. Hyla moorei. Some individuals were remapped to confirm the sequential nature of the changes described. The projection was retinotopic throughout regeneration. Multiunit receptive field sizes were initially large but progressively decreased to normal over approximately 30 days. The differences between these results and earlier studies are discussed. PMID- 6284309 TI - Sustained enhancement of evoked potentials following brief, high-frequency stimulation of the cerebral cortex in vitro. PMID- 6284310 TI - The prolonged hyperpolarizing afterpotential in an invertebrate photoreceptor: wavelength and ionic dependence. AB - A single electrode voltage clamp was used to examine the prolonged hyperpolarizing afterpotential (PHA) which accompanies photoconversion of a substantial fraction of rhodopsin (lambda max = 500 nM) to metarhodopsin (lambda max = 575 nM) in distal photoreceptor cells in the retina of the bay scallop, Pecten irradians. The PHA appears to result from a persistent light-activated outward K+ current passing through the same channels responsible for the normal receptor potential in these cells. PMID- 6284311 TI - Modulation of the histamine-induced inhibitory response in an identified Onchidium neuron by cyclic nucleotides. AB - Intrinsic beating activity in the identified molluscan neuron Be-1 can be inhibited for a relatively long time by a short application of histamine. Cyclic AMP enhanced both the amplitude and duration of this inhibitory histamine response (H2-response), while cyclic GMP depressed the H2-response. However, when only these nucleotides were applied, they produced no significant, change in the beating spike activity. PMID- 6284312 TI - Histamine-sensitive adenylate cyclase in monkey brain. PMID- 6284313 TI - Effects of 4-aminopyridine on synaptic transmission in the cat spinal cord. AB - An analysis was made of effects of 0.1-1.0 mg/kg 4-aminopyridine (4-AP) i.v. on excitatory and inhibitory spinal reflex pathways in lightly anaesthetized or decerebrated cats. The effects appeared within the first minutes of the injection, reached maximum after about 10-15 min and remained stable during at least several hours. 4-AP enhanced the following synaptic actions on motoneurones: monosynaptic excitation from Ia afferents and descending tracts, disynaptic and polysynaptic excitation from group Ib, group II, cutaneous and high threshold muscle afferents, disynaptic inhibition from Ia and Ib afferents and recurrent and polysynaptic inhibition from different afferents. 4-AP also increased primary afferent depolarization and excitation of ascending tract cells by peripheral stimuli. In the case of the disynaptic inhibitory pathways it has been shown that 4-AP may enhance the excitation of the interposed interneurones but it also increases the action of these interneurones on the motoneurones; monosynaptic inhibition evoked in motoneurones by electrical stimulation of the axons of the inhibitory interneurones was used as a test response in these experiments. No indications were found of direct effects of 4-AP on excitability of afferent fibres or motoneurones to electrical stimuli. No systematic changes were either found in the membrane potential of motoneurones or in the duration of action potentials of these neurones or primary afferents. It is therefore concluded that small doses of 4-AP enhance synaptic transmission in the spinal cord by an action at a presynaptic level. PMID- 6284314 TI - The corticotrigeminal projection in the cat. A study of the organization of cortical projections to the spinal trigeminal nucleus. AB - The projection from the cerebral cortex to the spinal trigeminal nucleus has been studied light microscopically in adult cats. Both orthograde degeneration and orthograde intra-axonal labeling techniques have been applied. Our results indicate that the projection from the coronal gyrus (face area of primary somatosensory cortex) to the spinal trigeminal complex is somatotopically organized. In subnucleus caudalis this somatotopy is organized dorsoventrally and appears to match the somatotopic distribution of the divisional trigeminal afferents. Hence cortical fibers originating from the posterior coronal gyrus (upper representation) project ventrolaterally into caudalis where division I trigeminal afferents terminate. Likewise cortical fibers from the anterior coronal gyrus (jaw and tongue representation) terminate dorsomedially in caudalis to overlap with division III trigeminal afferents. In contrast, the distribution of corticofugal afferents to the rostral spinal trigeminal subnuclei (pars interpolaris and oralis) is organized mediolaterally. Therefore in these subnuclei the cortical projection does not appear to overlap the dorsoventral lamination of the divisional trigeminal afferents. In addition, our results suggest that the cortical projection to subnucleus caudalis includes fibers which terminate in the marginal zone (lamina I) and its extensions into the spinal trigeminal tract (the interstitial cells of Cajal). We have been unable to document a projection from the proreate gyrus to the spinal trigeminal complex. PMID- 6284315 TI - Elevated divalent cation concentration decreases potassium-induced depolarization of bullfrog primary afferent fibers. PMID- 6284316 TI - [3H]Etorphine binding activity in early chick embryos: brain and body tissue. AB - Stereospecific [3H]etorphine binding has been detected in chick embryos as early as day 4 of incubation in both brain and body tissue. By day 10 of incubation [3H]etorphine stereospecific binding activity is not detectable in nonneuronal tissue. The ubiquitous opiate binding sites early in embryogenesis are high affinity and respond to ion and GTP regulation in a manner similar to adult brain tissue. We interpret our observations to indicate all embryonic cells prior to cell differentiation contain opiate receptors. Therefore, we propose that opiate receptors play a dual role; one function early in embryogenesis not associated with neurotransmitter regulation, and another function later in embryonic development and in the adult: the classical neurotransmitter regulatory function. PMID- 6284317 TI - Blood glucose profiles in surgically prepared flaxedilized and anesthetized rats. AB - Blood glucose profiles were monitored over an extended period of time in surgical rat preparations which included femoral artery and vein cannulations, tracheotomy, carotid artery cannulation and a craniotomy to expose the brain. Rats were under pentobarbital anesthesia, urethane anesthesia or Flaxedil neuromuscular blockage and local anesthesia, Fasted and unfasted rats were compared. These profiles are a foundation for studies of the central nervous system control of feeding behavior and metabolic homeostasis and of hypothalamic glucoreceptors, for which knowledge of the baseline blood glucose profile is particularly important. Such studies utilize the surgical preparations examined here. The importance of rapid "on the spot" "moment to moment" monitoring of blood glucose during the course of an experiment and adjustment of the experiment accordingly is demonstrated. Methods for such monitoring are evaluated in comparison with the standard enzymatic fluorometric assay for glucose. The YSI Glucose Analyzer seems to be a suitable one and is feasible for physiological and psychological laboratories without biochemical expertise. PMID- 6284318 TI - delta 9-THC and 17-beta-estradiol in hippocampus. AB - Electrophysiological field potentials recorded from in vitro hippocampal slice preparations show dose-dependent differences in response to 17-beta-estradiol (E2) and delta-9-tetrahydrocannabinol (THC) added to the incubation medium. Using a wide range of doses (1 pM-10 nM), it was found that mid-range concentrations of estradiol (100 pM) and THC (10 pM) tended to increase field potentials in CA1 of rodents. Higher dose levels of each agent were found to depress neuronal activity. In the context of prior findings, these results suggest that the two compounds share a common mechanism of action in the hippocampus. PMID- 6284319 TI - [Cytochemical localization of K+-p-NPPase in the myocardium of rats (author's transl)]. PMID- 6284320 TI - [Biochemistry of a voltage-sensitive sodium channel structure, mechanism and differentiation]. PMID- 6284322 TI - [Multiple opiate receptor sites in the rabbit thalamus and cerebellum: preliminary characterization (author's transl)]. AB - Inhibition of 3H-etorphine binding by D-Ala2, D-Leu5-enkephalin, a delta agonist and by morphiceptin, a mu agonist, reveals the existence of at least two classes of binding sites for the tritiated oviparine in membranes from the Rabbit thalamus and cerebellum. In the thalamus, approximately 50% of these sites appear to be mu receptor sites whereas the other half is neither of the mu nor of the delta subtype. In the cerebellum, at least 80% of the 3H-etorphine binding sites would belong to the mu subtype. This result may be related to the fact that the rabbit cerebellum contains appreciable quantities of methionine-enkephalin but little if any leucine-enkephalin. PMID- 6284321 TI - [Partial purification and characterization of nuclear DNA-topoisomerases from trypanosomes (author's transl)]. AB - We have prepared an extract presenting two types of topoisomerase activity from a nuclear fraction of Trypanosoma cruzi. The first one was revealed by the relaxation of supercoiled DNA molecules and was independent of ATP. Mg++ enhanced the reaction, without being necessary. Activity was observed between pH 5.4 and pH 8.5. The second activity was revealed by the formation of catenated molecules in the presence of ATP. PMID- 6284323 TI - Analysis of ovine colostrum to detect antibody against progressive pneumonia virus. AB - Immunoglobulins were isolated and purified from the colostrum and serum of progressive pneumonia virus infected sheep and also from non-infected control sheep. Four classes of immunoglobulins were isolated from sheep colostrum (IgG1, IgG2, IgA and Ig10s). Three classes of immunoglobulins were isolated from sheep serum (IgG1, IgG2 and IgM). Low levels of virus neutralizing activity were demonstrated only in the whole serum and purified serum IgG1 preparations. No complement fixing activity was detected in any of the antibody preparations from colostrum. PMID- 6284324 TI - Pseudorabies virus antibodies in swine slaughtered in Iowa. AB - Sera from butcher swine (1,246 total) were evaluated qualitatively by the microimmunodiffusion test and quantitatively by the virus neutralization test for antibody to pseudorabies virus. Ten percent of the sera had antibody to pseudorabies virus. Follow-up contact with veterinarians whose clients included the farms from which the positive swine originated revealed that few feeder swine are vaccinated against pseudorabies and that most infections with pseudorabies virus are subclinical. PMID- 6284325 TI - Types of avian infectious bronchitis strains isolated in Quebec. AB - Between 1976 and 1980, 24 isolates of infectious bronchitis virus were obtained from Quebec flocks. The serological classification of these isolates was demonstrated by cross neutralization tests using antisera to 13 different reference virus strains. Of the 24 isolates, ten were identified as Connecticut, six Holland and one SE-17 types. Seven strains did not react with any of the specific antisera. PMID- 6284326 TI - Isolation of an exotic serotype of bluetongue virus from imported cattle in quarantine. AB - In 1980, 60 zebu cattle from Brazil were admitted into quarantine in Florida for 150 days. During the 30 days between their last test in Brazil and their first test in Florida, four animals developed antibody to bluetongue virus detectable by agar gel immunodiffusion test. Within 62 days after arrival in Florida, three more seroconverted and one more was positive by the 86th day. Virus neutralizing titers of serums from the first four cattle were highest against bluetongue virus serotype 4 and 20; both of these serotypes are exotic to the United States. A bluetongue virus serotype 4 was isolated from one of these animals. The eight positive reactors were slaughtered; the other 52 cattle, which did not develop detectable antibody titers to bluetongue virus, were released into the United States. PMID- 6284327 TI - Correlation of hematological changes and serum and monocyte inhibition with the early suppression of phytohemagglutinin stimulation of lymphocytes in experimental infectious bursal disease. AB - Several experiments were conducted to study the mechanism of infectious bursal disease virus induced suppression of phytohemagglutinin stimulation of peripheral blood lymphocytes. Infectious bursal disease virus inoculation of one week old chicks resulted in significant suppression of phytohemagglutinin stimulation during the first three days after inoculation as demonstrated by a whole blood assay. Mild thymic necrosis was seen on day 3. Hematological changes during this time consisted of increased numbers of circulating lymphocytes and monocytes in infected chickens. Absolute monocyte counts remained elevated even after phytohemagglutinin stimulation had returned to normal. Furthermore, even after a 72.3% reduction in the monocyte population in leukocyte preparations, there was still marked viral induced suppression of phytohemagglutinin stimulation. An elevation in the absolute number of circulating large immature lymphocytes correlated with suppression of phytohemagglutinin stimulation. Sera from infected and control chickens depressed phytohemagglutinin stimulation of lymphocytes from control chickens at the 5 and 10% concentration. At the 1% concentration, inhibiton by control sera was considerably less than the inhibition by infected sera. The relationship between these findings and the mechanism of viral induced suppression of T-lymphocyte function is discussed. PMID- 6284328 TI - Enzyme linked immunosorbent assay (ELISA) for the detection of antibodies to porcine cytomegalovirus. AB - The ELISA and indirect immunofluorescence test were compared on 56 porcine sera which were tested for antibodies to porcine cytomegalovirus. Viral antigens were prepared in cells of a pig fallopian tube line. The ELISA was found to be a sensitive reproducible and practical test to measure specific antibodies to this infection. PMID- 6284329 TI - Sensitivity of seven different types of cell cultures to three serotypes of foot and-mouth disease virus. AB - The ability of bovine tongue origin foot-and-mouth disease virus serotypes A, O and C to replicate in seven different types of cell cultures was studied. Primary and secondary calf thyroid cells were equivalent in susceptibility to bovine kidney cell cultures passaged up to five times. Calf thyroid cells lost their susceptibility after two passages. Cryopreserved bovine kidney cell cultures passaged three and four times were equivalent in susceptibility to sensitive calf thyroid and bovine kidney cells. Susceptibility to foot-and-mouth disease virus serotype C was most variable among the cells tested. Lamb testicle and porcine kidney cells were susceptible to foot-and-mouth disease virus while goat and calf testicle and calf lung cells were refractory. PMID- 6284330 TI - Hepatocellular adenocarcinoma in a white-tailed deer (Odocoileus virginianus). AB - A white-tailed deer (Odocoileus virginianus), shot during the 1978-79 New Jersey hunting season, was presented with an enlarged, multinodular liver and numerous skin growths. The skin lesions were found to be fibromas and the liver tumor was identified as a hepatocellular adenocarcinoma, a rare neoplasm, not only in deer but all wild animals. PMID- 6284331 TI - Resistance of Listeria to cefotaxime and moxalactam. PMID- 6284332 TI - Benign and malignant fibrous histiocytomas of the soft tissues: functional characterization of the cultured cells. AB - Cell cultures were carried out from fresh tumor tissues obtained from seven cases of histiocytic tumors of the soft tissues including four of malignant fibrous histiocytoma, two of benign fibrous histiocytoma, and one of giant cell tumor of tendon sheath. The cultured cells were studied by light and electron microscopy, rosette formation for surface receptors, immune phagocytosis, and enzyme cytochemistry. The culture of each tumor revealed a mixture of histiocyte-like cells, fibroblast-like cells, amd intermediate forms. Bizarre giant cells were numerous in the early cultures of malignant fibrous histiocytoma, but they were absent in the cultures of benign fibrous histiocytoma and giant cell tumor of tendon sheath. In all the seven cases the cultured cells exhibited several features characteristic of histiocytes: (1) surface receptors for the Fc-portion of immunoglobulin G; (2) surface receptors for the third component of complement; (3) immune phagocytosis of opsonized erythrocytes; and (4) lysosomal enzymes including alpha-naphthyl butyrate esterase, and acid phosphatase. These findings suggest that the histiocytic tumors of the soft tissues in discussion actually comprise the cells that have the functional property of histiocyte. The tumor cells in the surgical specimens presented essentially the same ultrastructural and enzyme histochemical characteristics as those of the cultured cells. PMID- 6284333 TI - Autopsy findings in 154 patients with germ cell tumors of the testis. AB - Autopsy findings are reviewed in 154 patients treated for germ cell tumors of the testis. Of the patients with apparently pure seminoma, 44% had autopsy evidence of nonseminomatous metastases. For all tumor types, the most common sites of distant metastasis were lung (89%), liver (73%), brain (31%), and bone (30%). There was a high incidence of brain metastases in choriocarcinoma and of bone metastases in seminoma. Brain, liver, and bone metastases were late occurrences in the course of the disease and were almost always associated with involvement of other sites. Recurrences in the retroperitoneal area after lymph node dissection occurred mainly in those who had had retroperitoneal lymph node metastases. No difference in site or frequency of metastases was apparent in autopsied patients treated before or after introduction of platinum containing regimens. Respiratory failure, secondary to lung metastases, was the most common cause of death. Of the autopsied patients, 6% died of iatrogenic causes. PMID- 6284334 TI - Cannabis and cancer chemotherapy: a comparison of oral delta-9-THC and prochlorperazine. AB - Delta-9-tetrahydrocannabinol (THC) and prochlorperazine (Compazine) were found to be equally efficacious in reducing nausea and vomiting associated with cancer chemotherapy across a wide range of chemotherapeutic regimens and tumor types. Both drugs were administered orally one hour before chemotherapy, then every four hours for a total of four doses. Compazine was administered in a fixed dose of 10 mg; THC was administered by body surface area (BSA): BSA less than 1.4 m2 = 7.5 mg; BSA 1.4-1.8 m2 = 10- mg; and BSA greater than 1.8 m2 = 12.5 mg. Two hundred and fourteen subjects (75% of whom had previously received Compazine with varying results) were evaluated employing a double-blind, crossover design. Additional parameters evaluated were study drug effects on appetite, food intake, mood, activity, relaxation, interaction, and concentration. There were significant drug effects with THC: less ability to concentrate (P less than 0.01), less social interaction (P less than 0.05), and less activity (P less than 0.05). There were no significant differences between the two drugs in the level of food intake or appetite. Patients of all ages did equally well on both drugs. Neither past marijuana use nor past Compazine use were related to study the drug efficacy. Those patients who correctly identified their THC cycle did better on THC versus those who could not correctly identify which antiemetic they had received (P less than 0.05). There were more drug-related effects associated with THC, but these did not reduce the patients' preference for the drug, and were associated with nausea reduction (P less than 0.05). PMID- 6284335 TI - Cisplatin, adriamycin, and etoposide (CAV) for remission induction of small-cell bronchogenic carcinoma. AB - Chemotherapy with a combination of cisplatin (60 mg/m2), Adriamycin (45 mg/m2), and etoposide (120 mg/m2 X 3) (CAV) has been evaluated in 36 patients with small cell bronchogenic carcinoma (SCBC) after two full courses. The complete response (CR) rate was 23% in patients with extensive disease and 64% in patients with limited disease; the partial remission (PR) rate was 59% in patients with extensive disease and 22% in those with limited disease after two cycles (six weeks) of therapy. Patients with CR survived significantly longer than patients with PR (P = 0.02). Side effects were acceptable and consisted mostly of nausea, vomiting, alopecia, and myelosuppression. Thirteen patients were included into a "late intensification" program that was performed with increased doses of CAV regimen used for remission induction. This intensification of chemotherapy was carried out in a protective environment and with autologous bone marrow transfusion. In two patients with PR, CR could be obtained after late intensification and in one patient whose disease was progressing, PR had been achieved. However, excessive extramedullary toxicity of the late intensification regimen, consisting of mucositis, suggested that CAV does not appear to be the optimal therapy for further intensification. PMID- 6284336 TI - Acute leukemia following intensive therapy for small-cell carcinoma of the lung. PMID- 6284337 TI - Ultrastructure of fibrolamellar oncocytic hepatoma. PMID- 6284338 TI - Gynandroblastoma: its ultrastructure. AB - Gynandroblastoma is a rare, sex-cord stromal tumor of the ovary that shows morphologic evidence of female and male differentiation. Such a tumor produced masculinization in a 24-year-old woman, whose symptoms disappeared following removal of the tumor. By electron microscopy, the granulosa cell nests displayed Call-Exner (CE) bodies of the hyaline type composed of multiple layers of basal lamina resembling CE bodies of the normal graafian follicle. In contrast, CE bodies of a classic granulosa theca cell tumor were of the spongiform type, consisting of a space limited by a single basal lamina containing altered granulosa cells and cell processes. Both types of CE bodies are believed to arise following secretion by and/or degeneration of granulosa cells, the variation in morphology between the two resulting from differences in amounts of basal lamina deposited. The tubular components of the tumor resembled more closely the rete ovarii than did Sertoli cells, and its proposed that such structures be called by the alternate and less specific term "androblastoma." The identity of Leydig cells was established by demonstrated of intracytoplasmic Reinke crystals. Despite a difference in architectural pattern, there was a close ultrastructural resemblance between the different sex-cord components of the gynandroblastoma. PMID- 6284339 TI - Histopathologic studies of basal cell adenoma of the parotid gland. AB - Five-hundred-thirty-one cases of primary epithelial tumors of the parotid gland were examined. The incidence of basal cell adenomas was 7.5% (40 cases). The tumors could be classified histologically into these subtypes: basal cell (21/40; 42.5%); tubular, (13/40; 32.5%); trabecular (4/40; 10.0%); and papillary (2/40; 5.0%). The pattern of classic basal cell adenoma predominated in the basal-cell type but was also seen in varying degrees in the latter three types with a reciprocal transition. Accordingly, it could be suggested that tubular, trabecular, and papillary types are variants of basal cell adenoma. The higher incidence of basal cell adenomas in our survey could be accounted for by this categorization. Histologically, basal cell adenomas presented various features; (1) cystic formation (26/40; 65.0%), being most frequently seen in tubular and trabecular types; (2) adenoid cystic pattern (4/40; 10.0%); (3) elastosis in the stromal tissues (2/40; 5.0%). Basal cell adenomas were clinicopathologically compared with 291 cases of pleomorphic adenomas. Basal cell adenomas were seen more often in female patients, (60.0%), as were pleomorphic adenomas (68.4%). They were observed more frequently in patients over 50 years of age and the average was ten years higher than for pleomorphic adenoma. The tumor size tended to be smaller (below 2 cm at the greatest diameter) than pleomorphic adenomas. PMID- 6284340 TI - Brain metastasis in the natural history of small-cell lung cancer: 1972-1979. AB - The records of 134 patients with small-cell lung cancer who had been seen at Henry Ford Hospital between 1972-1979 were reviewed with emphasis on brain metastasis. Brain metastasis developed in 28% of the patients; 21% had brain metastasis at the time of diagnosis and in 7% it developed later (de novo). In 9%, the chief complaint was related to brain metastasis. Brain metastasis in only two of 109 patients developed after diagnosis (de novo) yet prior to initiation of prophylactic brain radiation. The frequency of de novo brain metastasis in those treated with intensive combination chemotherapy and those treated by other means was similar. Of the patients with brain metastasis, 73% died from other causes. Isolated brain metastasis in the absence of progressive systemic disease was rare. PMID- 6284341 TI - Primary gastric plasmacytoma with massive intracytoplasmic crystalline inclusions: a case report. PMID- 6284343 TI - Malignant fibrous histiocytoma of the mediastinum. PMID- 6284342 TI - Small-cell carcinoma of the endometrium: light microscopic and ultrastructural study of a case. PMID- 6284344 TI - Minute gastric cancers less than 5 mm in diameter. AB - Minute gastric cancers with maximum dimensions of less than 5 mm were studied clinicopathologically. There were 49 intramucosal cancers among 46 patients and nine submucosal cancers among nine patients. No lymph node metastasis was found. Macroscopically, eight (13.8%) were evaluated, 12 (20.7%) were flat, and 38 (65.5%) were depressed. Most submucosal cancers were of the depressed type (8/9, 88.9%). Classification of cancers according to association with other large cancers into single group (22 cases) and multiple group (33 cases) revealed that (1) the majority of the minute gastric cancers (20/22, 90.9%) in the single group were the depressed type and (2) submucosal cancers in the single group were 8/22, accounting for 36.4%, a much higher incidence as compared with 1/36 (2.9%) in the multiple group. These facts indicate that gastric cancers should be detected when they are about 5 mm in maximum dimension and before they invade beyond the submucosal layer, especially in single and depressed type. PMID- 6284345 TI - Combination chemotherapy (vincristine, adriamycin, cyclophosphamide, and 5 fluorouracil) in the treatment of children with malignant hepatoma. AB - Members of Childrens Cancer Study Group and the Pediatric Division of the Southwest Oncology Group conducted a study of chemotherapy for children with malignant liver tumors. All patients received vincristine, cyclophosphamide, Adriamycin and 5-fluorouracil in 6 weekly cycles for one year. Surgical resection and irradiation were employed when indicated. Between January 1976 and August 1978, 62 patients were entered on study; one was rejected for a protocol error, and ten had inadequate trials of chemotherapy, dying within one month of entry. The median time on study for all patients was 12 months. Twenty-four patients had no measurable disease following surgical treatment and chemotherapy was employed as adjuvant treatment; 20/24 (83%) remain relapse-free from 8-42+ months, (median, 30 months). In 27 patients, residual measurable disease was available to determine the response to chemotherapy. The response rate was 12/27 (44%), lasting 3-45 months (median, 18 months). The median follow-up of all survivors is 30 months. Hematologic toxicity was significant, particularly during the initial courses of chemotherapy; 28/57 patients developed severe toxicity which was fatal in three. The results from the current study were compared to those from a previous one initiated in 1972, in which actinomycin D, vincristine, and cyclophosphamide were given in sequence, one during each month for one year. Although the population of the two studies was not identical, there was a difference in the response rates (P = 0.02), relapse-free interval (P = 0.008), and survival (P = 0.003). The most striking improvement was seen in the patients with Group I disease, there were 7/11 relapses in the first study and 1/16 in the current one. PMID- 6284347 TI - A rational approach to the radiologic evaluation of children with Wilms' tumor. PMID- 6284346 TI - Multifocal relapse after concurrent chemotherapy and radiotherapy of small cell lung cancer. AB - Twenty-one patients with small cell lung cancer (SCLC) were treated with cyclophosphamide (1250 mg/m2), Adriamycin (40 mg/m2), vincristine (2 mg) every three weeks. Thoracic radiotherapy (3000 rad: 10 or 15 fractions) began four weeks after starting chemotherapy. Patients with brain metastases received cranial irradiation. Thirteen of 19 evaluable patients responded to therapy (four complete responses). Sixteen of 19 had significant intrathoracic response (eight complete). Of the three patients without an intrathoracic tumor response, two had simultaneous progression in systemic locations and only one had intrathoracic progression preceding systemic progression. Intrathoracic relapse preceded systemic relapse in two patients, was simultaneous with it in six and followed systemic relapse in four others. Therefore only three patients had intrathoracic tumor progression or relapse preceding systemic progression or recurrence. The more intensive course of radiotherapy was significantly better in preventing intrathoracic tumor progression. Although intrathoracic tumor control is important, primary failure of therapy or relapse appears to be multifocal. Future attention must be directed toward control of multiple potential sites of relapse. PMID- 6284348 TI - NMR relaxation times of water protons in human colon cancer cell lines and clones. AB - Established lines of human colon cancer cells from several sources (LS180, LS174T, HT29, SW480, SW1345) had water proton nuclear magnetic resonance (NMR) spin-lattice relaxation times (T1) of 460 +/- 45 msec to 982 +/- 9 msec and spin spin relaxation times (T2) of 83 +/- 6 msec to 176 +/- 6 msec. Two clones derived from single cells of line LS174T were similar in T1 and T2 to the parent line. Differences among the cell lines were not totally a function of cellular hydration. Normal adult and fetal human primary colon cells were wetter and had higher T1 and T2 values than established cell lines. Relaxation times in this study substantiate variations seen for human colon tumors in earlier studies. Established cell lines maintained water relaxation times similar to tumor tissue values. Along with other morphological and biochemical criteria, the relaxation times suggest that these established human colon cancer cell lines may serve as a good experimental model for the study of human colon cancer. PMID- 6284349 TI - Genetic studies on hydatidiform moles. I. The origin of partial moles. AB - A study was designed to investigate the genetic origin of hydatidiform moles. Fifty-nine specimens were obtained and, on a histological basis, separated into two entities: complete and partial. The study of the genetic origin of the 15 partial moles, using cytogenetic and biochemical markers, is described. All the partial moles examined cytogenetically were triploid. One had 71 chromosomes. The sex chromosome complements of seven cases were six XXY and one XXX. Origin by dispermy was possible in seven cases and was proven in four. With the use of biochemical markers a maternal contribution was identified in three cases, and the isoenzyme pattern suggested a trisomic state for at least one locus in four cases. The mechanism of origin of partial moles was compared with spontaneously aborted and liveborn triploids. All the patients were followed up for at least 9 months, and none required treatment for persistent trophoblastic activity. PMID- 6284350 TI - Reversal of the promotional effect of 12-O-tetradecanoylphorbol-13-acetate on morphological transformation of hamster embryo cells by glucocorticoids. AB - The effects of glucocorticoids and cholesterol on morphological transformation have been studied using hamster embryo cells. The cells were exposed sequentially to benzo[a]pyrene (BP) (3 days) and the tumor promotor 12-O-tetradecanoylphorbol 13-acetate (TPA) (4 days) before scoring for morphologically transformed colonies. Dexamethasone and hydrocortisone strongly inhibited the formation of morphologically transformed colonies when applied to the cells in the second period of exposure together with TPA, but had no effect when present with BP during the first period. Corticosterone had only a weak effect, while cortisone had no effect on the transformation frequency. Cholesterol gave rise to an enhanced number of transformed colonies. Dexamethasone, being the most potent compound tested, inhibited morphological transformation by more than 50% when present in a concentration of 0.25 nM. The finding that dexamethasone reduced the frequency of transformation even when added only 6 h prior to staining, indicates that the glucocorticoids may also reduce the transformation frequency by reversing morphologically transformed cell colonies to a normal appearance. PMID- 6284351 TI - Regulation of cell division in a human glioma cell clone by arachidonic acid and alpha-tocopherolquinone. AB - A human glioma cell clone (12-18 CV), derived from a previously characterized glioblastoma multiforme cell line, was established in culture. Lipid peroxidation (thiobarbituric acid test) occurred when either 8,11,14-eicosatrienoic acid or 5,8,11-eicosatetraenoic acid (arachidonic acid) was added to the cells in culture. The extent of lipid peroxidation was similar in fetal brain cells (CH II) treated with these polyunsaturated fatty acids. The antioxidant, alpha tocopherolquinone, inhibited lipid peroxidation in both the glioma cell clone and fetal brain cells. Arachidonic acid significantly reduced cell division in both the glioma cell clone and fetal brain cells. alpha-Tocopherolquinone restored cell division to control levels in both cultures. These data show that cells from a tumor clone retain the capacity for lipid peroxidation. Furthermore, cell division in a tumor clone is correlated with lipid peroxidation in the same way that cell division in other cell lines is correlated with lipid peroxidation. PMID- 6284352 TI - Characterization of an amino acid fucoside of normal and SV40-transformed human embryonic lung cells. AB - The incorporation of radioisotopically labeled fucose into a prominent fucosylated component, i.e., fucose-labeled amino acid fucoside 4c (FL4c), of human embryonic lung cells is markedly decreased in cell lines derived from human tumors. In the current study, we have extended the above observations by examining more closely related normal and transformed human cells, e.g., Wi38 cells and SV40-transformed Wi38 cells. We have found that the level of radioisotopically labeled fucose incorporated into FL4c of the SV40-transformed human embryonic lung cells is dramatically reduced as compared to their normal counterpart cells. Additionally, we have chemically characterized FL4c. FL4c was purified from monkey tissues using a combination of gel filtration chromatography, ion-exchange chromatography, and preparative thin-layer chromatography. Analysis of this material was accomplished by amino acid analyzer and by gas-liquid chromatography; fucose, glucosamine, and aspartic acid in molar ratios of approximately 1.0:1.0:1.0 were observed. Furthermore, alpha-L fucosidase treatment of [3H]fucose-labeled FL4c revealed that the fucose was alpha-linked and in a terminal position. Similar treatment of [3H]glucosamine labeled FL4c yielded a component that cochromatographed with N acetylglucosaminylasparagine. The combined results of the structural studies are consistent with the structure of FL4c being alpha-fucosyl-N acetylglucosaminylasparagine. PMID- 6284353 TI - Purification and some properties of a deoxyribonucleoside kinase from L1210 cells. PMID- 6284354 TI - Morphologic and neoplastic transformation of Syrian hamster embryo fibroblasts by diethylstilbestrol and its analogs. AB - A mammalian cell culture system using normal, diploid Syrian hamster embryo fibroblasts was used as a model to study the ability of diethylstilbestrol (DES) and related compounds to induce neoplastic transformation. Like benzo(a)pyrene, a known chemical carcinogen, DES (0.01 to 10 micrograms/ml) induces morphological transformation of Syrian hamster embryo fibroblasts in vitro; the transformed colonies were indistinguishable from colonies of benzo(a)pyrene-transformed cells. The morphologically transformed colonies derived from either benzo(a)pyrene- or DES-treated cells were tumorigenic when injected into newborn hamsters. At doses of 0.01 to 1 microgram/ml for 48 hr, DES did not inhibit or stimulate cell proliferation. Structural analogs of DES were also tested in the Syrian hamster embryo transformation system. Like DES, tetrafluorodiethylstilbestrol, dimethylstilbestrol, and cis,cis-dienestrol morphologically transformed Syrian hamster embryo cells. The transformation frequency of dimethylstilbestrol was much less than that of DES, tetrafluorodiethylstilbestrol, or cis,cis-dienestrol. Hexestrol, dimethoxydiethylstilbestrol, and trans,trans-dienestrol did not transform these cells. No correlation could be demonstrated between reported estrogenic potency of the compounds and their cell-transforming capacity. Structure-activity relationships developed for these chemicals suggest that metabolism via specific pathways plays a role in DES-induced cell transformation. PMID- 6284355 TI - Induction of retrovirus gene expression in mouse cells by some chemical mutagens. AB - Cell cultures derived from a variety of mouse strains were compared for their relative capacity to be induced to express endogenous retrovirus proteins by exposure to 5-iododeoxyuridine under optimized experimental conditions. Induction frequencies varied between 6.0 x 10(-1) and 1.7 x 10(-2) with AKR cells showing the highest capacity and C57BL/6 x C3H F1 cells the lowest. Virus expression was induced in AKR cells with other chemical mutagens of the polyaromatic hydrocarbon [benzo(a)pyrene and phenol, diol, and epoxide metabolites] and nucleoside analog classes, but alkylating agents were inconsistent or failed to induce. Considerable differences in the efficiency of induction were seen between various halogenated nucleosides, while under these conditions the nucleoside 5 azacytidine induced greater than 90% of AKR cells at a concentration of 2 to 4 micrograms/ml. The high frequency of induction by 5-azacytidine, relative to other nucleoside analogs, and the absence of induction by other mutagens further indicate that endogenous virus induction occurs via nonmutagenic mechanisms and that some mutagens may also affect regulatory functions independent of their mutagenic action. PMID- 6284356 TI - Effect of phorbol myristate acetate and a lymphokine on cyclic 3':5'-guanosine monophosphate levels and proliferation of macrophages. AB - The tumor promoter phorbol myristate acetate (PMA) was compared to a lymphokine macrophage mitogenic factor (MMF) for its ability to induce replication of guinea pig peritoneal and alveolar macrophages. Like MMF, PMA induces DNA synthesis of both cell populations with peak thymidine incorporation at 72 hr of culture. Optimal concentrations of PMA for the peritoneal and alveolar cells were 1.6 x 10(-7) and 1.6 x 10(-9) M, respectively. The magnitude of the effect is slightly less than MMF but greater than that of phytohemagglutinin or concanavalin A. Indomethacin added to inhibit prostaglandin synthesis potentiates the effects of MMF but has little effect on the actions of PMA and the other mitogens. Potentiation by indomethacin of the effects of PMA on the peritoneal cell was observed only at the suboptimal concentration of PMA (1.6 x 10(-8) M). By adherence criteria and density gradient fractionation, the cell responding to PMA is confirmed to be the macrophage. Cell counts and nuclear radioautography confirm that replication in this system is reasonably well reflected by thymidine incorporation. The effects of PMA and its analogs as macrophage mitogens correlate with their tumor-promoting effects. Both PMA and MMF induce early increases in peritoneal macrophage levels of cyclic 3':5'-guanosine monophosphate without changes in the levels of cycles 3':5'-adenosine monophosphate. These studies indicate that PMA offers a useful probe of macrophage function. PMID- 6284357 TI - Modification of the sensitivity and repair of potentially lethal damage by diethyldithiocarbamate during and following exposure of plateau-phase cultures of mammalian cells to radiation and cis-diamminedichloroplatinum(II). AB - Diethyldithiocarbamate (DDC), a chelating agent known to reduce levels of superoxide dismutase and glutathione peroxidase, appears to protect irradiated monolayers of mammalian cells when present for 1 hr before and during irradiation. To examine a possible cause of this modification, the repair of potentially lethal X-ray damage was examined with and without the presence of DDC in the medium overlying the cells postirradiation. Although little repair was seen in full medium alone when DDC was added to the full medium, the amount of repair was comparable to that seen under optimum repair conditions, that is, in Hanks' balanced salt solution. The t 1/2 of the repair process in Hanks' balanced salt solution or in full medium with DDC added was comparable and of the order of 1 to 1.5 hr. The cis-platinum sensitivity of the monolayers is significantly modified by the addition of DDC, and the nature of the modification is dependent upon the time at which the DDC is added to the cells following initiation of cis platinum exposure. To investigate a possible reason for this protection by DDC, we examined the repair of potentially lethal cis-platinum damage in the cell monolayers. Minimal repair was noted in the presence of either Hanks' balanced salt solution or full medium, but when DDC was added to the full medium, the repair was tripled, and the t 1/2 of the repair process was approximately 2 hr. The ability of DDC to protect cells from exposure to both X-rays and cis platinum, together with its augmentation of repair of potentially lethal damage following exposure to each, has broad clinical application and is being actively explored in tumor-bearing mice. PMID- 6284358 TI - Phorbol diester and epidermal growth factor receptors in 12-O tetradecanoylphorbol-13-acetate-resistant and -sensitive mouse epidermal cells. AB - Several cell variants have been isolated from promotable mouse JB6 epidermal cells which are resistant either to mitogenic stimulation at quiescence or to promotion of anchorage independence by 12-O-tetradecanoylphorbol-13-acetate (TPA). Such resistant variants would be expected to lack one or more steps in the TPA response pathway leading to mitogenesis or promotion of tumor cell phenotype. This report is concerned with determining whether resistance is attributable to lack of receptors for phorbol diesters or epidermal growth factor (EGF, a potential mediator) or to absence of receptor down modulation following ligand binding. The results show that neither lack of phorbol diester receptors nor absence of down modulation can be demonstrated in the TPA-resistant variants. The phorbol ester binding affinity is also not altered in the resistant variants. The presence of EGF receptors cannot be an absolute requirement for TPA promotion sensitivity since three of the TPA-promotable cell lines lack available EGF receptors. Lack of EGF receptors may account for TPA mitogen resistance in at least three of four resistant variants. The TPA-induced EGF binding decrease occurs in both sensitive and resistant variants. Thus, phorbol diester binding and receptor down modulation remain as possible required events in mitogenic and promotion responses to TPA. EGF receptors are clearly not necessary for TPA promotion of anchorage independence in JB6 cells but may mediate mitogenic stimulation of these cells by TPA. PMID- 6284359 TI - Effect of glucagon on amino acid transport and cyclic adenosine 3':5' monophosphate production in rat hepatoma cell line McA-RH 8994 in culture. PMID- 6284360 TI - CCNU, vincristine, methotrexate, and procarbazine treatment of relapsed small cell lung carcinoma. AB - Thirty-two patients with refractory relapsed small cell carcinoma of the lung (SCCL) were treated with a combination of CCNU (lomustine), vincristine, methotrexate, and procarbazine (COMP); 29 were evaluable for response. Nine patients (31%) had responses: five complete responses (CR) and four partial responses. Patients with CR had a median survival of 11 months (range, 51/2 141/2) from the start of COMP. Patients with less than CR had a median survival of approximately 3 months (range, less than 1-7). The comparison of CR versus less than CR is significant (P = 0.003). Patients achieving CR usually had limited disease and four of the five who achieved CR survived greater than 6 months. The regimen was well-tolerated, but myelosuppression was seen in all patients. COMP appears to be a useful combination in patients with relapsed SCCL. Aggressive retreatment should be considered in relapsed patients with this disease since some may achieve a second CR, with the associated potential survival benefit. PMID- 6284362 TI - Phase II trial of prednimustine in children with recurrent cancer: a Children's Cancer Study Group report. PMID- 6284361 TI - Phase II trial of doxorubicin therapy for advanced islet cell carcinoma. AB - Twenty evaluable patients with advanced islet cell carcinoma were treated with doxorubicin at a dosage of 60 mg/m2 every 3-4 weeks. Toxicity was clinically tolerable and characteristic of doxorubicin therapy. Although all patients had previously been treated with other chemotherapy regiments, four (20%) had objective therapeutic responses persisting for 2-301/2 months. Doxorubicin has definite therapeutic activity in islet cell carcinoma and is currently being evaluated in combination chemotherapy regimens. PMID- 6284363 TI - Extra-renal control of body potassium. PMID- 6284365 TI - [Glycides after prolonged fasting in rats (author's transl)]. PMID- 6284364 TI - [Symptomatic hepatic porphyria in workers exposed to free silicon dioxide (author's transl)]. PMID- 6284366 TI - [Monocyte isolation using Percoll. A comparison with the adherence method (author's transl)]. PMID- 6284367 TI - Globin pseudogenes. PMID- 6284368 TI - Genes of the major histocompatibility complex. PMID- 6284369 TI - A family of genes that codes for ELH, a neuropeptide eliciting a stereotyped pattern of behavior in Aplysia. AB - We describe a particularly advantageous experimental system for studying gene structure, expression and modulation in the nervous system. In the marine mollusc Aplysia, the bag cells, two discrete clusters of neurons, secrete a peptide of known behavioral function. This neuroactive peptide, egg-laying hormone (ELH), produces a characteristic and stereotypic behavioral repertoire, consisting first of a cessation of walking and inhibition of feeding, followed by head waving and egg laying. We have cloned the genes encoding ELH and characterized their organization and expression. At least five distinct genes for ELH exist within the chromosome. Sequence analysis of one recombinant clone unambiguously identifies a contiguous stretch of nucleotides that encodes the 36 amino acids of ELH. Transcription of this small multigene family results in the expression of at least five distinct RNA transcripts encoding ELH. The pattern of transcripts differs strikingly in different tissues: bag cells express three distinct mRNA species, whereas the atrial gland, a secretory reproductive gland, expresses two distinct mRNAs. Several other neuronal and nonneuronal tissues do not express ELH RNA. In vitro these mRNAs produce a series of long polypeptide precursors that must be processed to generate the active ELH peptide. This processing event is likely to generate several additional neuroactive peptides. Thus the same peptide, ELH, may be released in association with different combinations of other neuroactive peptides. The concept of combinatorial sets of neuropeptides, each bearing one overlapping peptide ELH, and each directing a differing pattern of behavior, greatly expands the information potential of a small set of genes. PMID- 6284370 TI - Critical sequences within mitochondrial introns: pleiotropic mRNA maturase and cis-dominant signals of the box intron controlling reductase and oxidase. AB - We have established the DNA sequence of nine yeast mutants that prevent the expression either of the split cytochrome b gene alone (five mutants) or of two split genes, the cytochrome b gene and the cytochrome oxidase subunit I gene (four mutants). All the mutations analyzed are localized in intron 14 of the cob box gene. We have extended the concept of the intron-encoded mRNA maturase, already described for intron 12, to the intron 14, and have adduced evidence that this box7 pleiotropic maturase is involved in the splicing of two distant gene transcripts. Such a process may constitute a regulatory mechanism that coordinates the expression of two structurally nonhomologous genes encoding two metabolically related enzymes. Analyses of cis-dominant mutations reveal the role of signal sequences in the recognition of the intron RNA sequences to be excised. These signal sequences are localized near the exon-intron boundaries (box1), or quite distant from the splicing sites, either in the blocked reading frame (box2) or in the open reading frame (box9) of the intron. We believe that for the last sequence, a ribosomal recognition of the box9 signal could be involved in a regulatory mechanism of the splicing of the pre-mRNA. PMID- 6284371 TI - Critical sequences within mitochondrial introns: cis-dominant mutations of the "cytochrome-b-like" intron of the oxidase gene. AB - We have established the DNA sequence of two cis-dominant mutations located in the fourth intron, a14, of the yeast mitochondrial gene oxi3. These mutations prevent the synthesis of subunit I of cytochrome oxidase. Both mutations affect a very short DNA sequence located several hundred base pairs from the intron-exon junctions. An identical sequence is found in the cob-box gene; and this sequence is critical for the excision of the cytochrome b intron. Our interpretation is that this short sequence represents a common signal that must be recognized by the box7-encoded mRNA maturase, in conjunction with the mitochondrial ribosome, to splice out the introns in the two nonhomologous genes, cob-box and oxi3. PMID- 6284372 TI - Generation of authentic 3' termini of an H2A mRNA in vivo is dependent on a short inverted DNA repeat and on spacer sequences. AB - We have determined what sequences are required to generate the authentic 3' termini of a sea urchin H2A histone mRNA. We have constructed a series of deletion and insertion mutants in the cloned histone repeat unit h22 of Psammechinus miliaris and have analyzed the transcripts of both wild-type and mutant DNAs produced in the frog oocyte. The protein-coding sequences of the H2A gene can be removed without any deleterious effects on transcription initiation or termination. A 12 bp deletion, which removes a highly conserved inverted DNA repeat immediately preceding the H2A mRNA 3' terminus, elicits read-through of the polymerase into the spacer DNA further downstream. However, the inverted repeat and the sequence coding for the 3' terminus of the mRNA are by themselves not sufficient to generate faithful 3' ends. Our data suggest that spacer sequences downstream of the 3' mRNA terminus are required as well. PMID- 6284373 TI - Differential expression of the members of the discoidin I multigene family during growth and development of Dictyostelium discoideum. AB - Discoidin I and II are lectins synthesized during the aggregation of Dictyostelium discoideum amoebae which may play a role in cellular cohesion. Discoidin I was thought to consist of two major polypeptides, but we show that there are three. The N-terminal amino acid sequence of the polypeptides has been predicted by determining part of the nucleotide sequence of their respective mRNAs. We obtained the nucleotide sequences by reverse transcription of the mRNAs, using as primers, fragments derived from the coding region of two cloned discoidin I sequences, and utilizing cross hybridization to the various mRNA species and differences in the length of their 5' noncoding regions to isolate fragments for DNA sequencing. We used primer extension to measure the relative concentration of the three major discoidin I mRNA sequences. We show that during development changes in the abundance of all three mRNA sequences occur coordinately. In cells growing in nutrient broth, however, only two of the three major discoidin I mRNA sequences accumulate, and if such cells are grown to a very high density, both sequences disappear. These results indicate that the coordination of discoidin I gene expression is not obligatory and that the members of this multigene family may differ in the mode of their induction during normal development. PMID- 6284374 TI - Membrane sites regulating developmental gene expression in Dictyostelium discoideum. AB - Postaggregative gene expression in Dictyostelium discoideum requires cell contact. Polyspecific monovalent antibodies (Fab) prepared from sera raised against membranes of aggregation- and postaggregation-stage cells were used to probe the cell interactions that induce rapid postaggregative synthesis of UDP glucose pyrophosphorylase. When cells of strain V12M2 were dissociated after 8 hr of development and replated in the presence of immune Fab, both reaggregation and pyrophosphorylase synthesis were blocked. Fab neutralized by incubation with EDTA high salt extracts of cells developed for 3 hr blocked pyrophosphorylase synthesis but not reaggregation. Therefore, some cell-surface components that regulate pyrophosphorylase synthesis (called E sites) are antigenically distinct from those required for reaggregation. The Fab provides a means to assay E sites during their purification. Addition of 10(-3) M cyclic AMP or cyclic GMP enabled the cells to bypass the blocking of E sites by Fab; pyrophosphorylase was synthesized in the absence of reaggregation. We hypothesize that E sites function by raising the level of intracellular cyclic AMP. PMID- 6284375 TI - A membrane glycoprotein that accumulates intracellularly: cellular processing of the large glycoprotein of LaCrosse virus. AB - The intracellular transport and certain posttranslational modifications of the large glycoprotein (G1) of LaCrosse virus (LAC) in BHK cells have been studied. G1 from released LAC virus was characterized by complex oligosaccharides (endo H resistant) and covalently attached fatty acid. Only a small fraction of total cellular G1 was present on the baby hamster kidney cell surface. Cell-surface G1 contained complex oligosaccharides, while total G1 in infected cells contained largely unprocessed (endo H-sensitive) oligosaccharides. In addition, cell G1 contained significantly less fatty acid than virion-associated G1. Pulse-chase experiments showed that the oligosaccharides of G1 were processed to the complex from much more slowly than the oligosaccharides of the vesicular stomatitis virus (VSV) glycoprotein (G). In addition, transit of LAC G1 to the cell surface and into extracellular virions was two to three fold slower than the transit of VSV G. Thus LAC G1 accumulates intracellularly and is only slowly processed by intracellular processing enzymes. Treatment with monensin caused accumulation in the cell of a form of G1 with partial sensitivity toward endo H, suggesting that monensin may act to inhibit the glycosylation process directly. PMID- 6284376 TI - The morphologic pathway of exocytosis of the vesicular stomatitis virus G protein in cultured fibroblasts. AB - Cultured fibroblasts were infected with vesicular stomatitis virus (VSV) and the pathway of exocytosis of G protein, the transmembrane glycoprotein of VSV, was followed by immunofluorescence and electron microscopy. G protein was detected within the endoplasmic reticulum, within smooth vesicles and stacks in the Golgi region and on the cell surface. No G protein was detected in the coated regions of the Golgi. Our data are consistent with the hypothesis that coated regions of the Golgi are involved in transfer of lysosomal enzymes and other substances to lysosomes and not in exocytosis. PMID- 6284377 TI - Stage-specific transforming genes of human and mouse B- and T-lymphocyte neoplasms. AB - DNAs of 20 B- and T-lymphocyte neoplasms of human and mouse origin induced transformation of NIH/3T3 cells with high efficiencies, indicating that these neoplasms contained activated transforming genes that were detectable by transfection. Analysis of the susceptibility of the transforming activities of lymphocyte-neoplasm DNAs to digestion with restriction endonucleases indicated that the same or closely related transforming genes were activated in independent neoplasms representative of the same stage of normal cell differentiation. However, different transforming genes were activated in neoplasms representative of different stages of normal B- and T-lymphocyte differentiation. These results indicate that specific transforming genes are activated in neoplasms of discrete stages of differentiation within these cell lineages. PMID- 6284378 TI - Genetic interactions in induction of endogenous murine leukemia virus from low leukemic mice. AB - The frequency of ecotropic murine leukemia virus (MuLV) production in cells induced with halogenated pyrimidines has been investigated in several low leukemic strains of mice. Very few BALB/c or C57BL/6 (B6) induced embryo cells produce MuLV; this low frequency increases 10 to 50 fold in cells of the BALB/c x B6 F1 hybrid. Data from back-crosses of the F1 hybrid to each parent and from BALB/c x B6 recombinant inbred strains indicate that the phenotype of enhanced MuLV production results from interaction of two unlinked loci, dominant (+/+) alleles of which are carried by either parent. Genetic tests with BALB/c x B6 recombinant inbred strains confirm this two-locus model. The loci are designated Inc-1 and Inb-1 to signify their phenotypic detection by induction and the BALB/c or B6 strain of origin, respectively. Examination of hybrids of BALB/c and of B6 with other strains indicates that strains related in pedigree to BALB/c carry Inc 1, whereas those related to B6 carry Inb-1. Identification of genetic loci that specifically interact to enhance MuLV production after exposure to halogenated pyrimidines indicates the existence of mechanisms that regulate the induction or intracellular expression of endogenous MuLV. PMID- 6284379 TI - Changes in amino-terminal sequences of pp60src lead to decreased membrane association and decreased in vivo tumorigenicity. AB - We have suggested previously that the amino-terminal 8 kilodaltons of pp60src may serve as a structural hydrophobic domain through which pp60src attaches to plasma membranes. Two isolates of recovered avian sarcoma viruses (rASVs), 1702 and 157, encode pp60src proteins that have alterations in this amino-terminal region. The rASV 1702 src protein (56 kilodaltons) and the 157 src protein (62.5 kilodaltons) show altered membrane association, and fractionate largely as soluble, cytoplasmic proteins in aqueous buffers, ion contrast with the membrane association of more than 80% of the src protein of standard avian sarcoma virus under the identical fractionation procedure. Plasma membranes purified from cells transformed by these rASVs contain less than 10% of the amount of pp60src found in membranes purified from cells transformed by Rous sarcoma virus or control rASVs. The altered membrane association of these src proteins had little or no effect on the properties of chick embryo fibroblasts transformed in monolayer culture. In contrast, rASV 1702 showed reduced in vivo tumorigenicity compared with Rous sarcoma virus or with other rASVs that encode membrane-associated src proteins. Rous sarcoma virus-induced tumors are malignant, poorly differentiated sarcomas that are lethal to their hosts. rASV 1702 induces a benign, differentiated sarcoma that regresses and is not lethal to its hosts. These data support the role of amino-terminal sequences in the membrane association of pp60src, and suggest that the amino terminus of pp60src may have a critical role in the promotion of in vivo tumorigenicity. PMID- 6284380 TI - Accessory cells in the in vitro generation of type C virus-specific T-killer lymphocytes. III. Two methods of antigen presentation of cytolytic T-lymphocyte precursors. PMID- 6284381 TI - Morphological and morphometrical differences between the binding sites of low- and high-density lipoproteins on the plasma membrane of cultured human fibroblasts. PMID- 6284382 TI - Aging of endothelium in culture: decrease in angiotensin-converting enzyme activity. PMID- 6284383 TI - Utilization of gamma-32P-GTP by ectoprotein kinase of 3T3 and SV40 3T3 cells. PMID- 6284384 TI - Caerulein-induced distribution of calmodulin, cGMP, and cGMP-dependent protein kinase in guinea pig pancreatic acinar cells. PMID- 6284385 TI - [Focal dermal hypoplasia syndrome (Goltz-Gorlin syndrome) (author's transl)]. PMID- 6284386 TI - [Pertussis (in clinical practice)]. PMID- 6284387 TI - [Pyrophosphate scan of the heart in cardiomyopathies (author's transl)]. PMID- 6284388 TI - Formation of mutagenic metabolites from benzo(a)pyrene and 2-aminoanthracene by the s-9 fraction from the liver of the Northern pike (Esox lucius): inducibility with 3-methylcholanthrene and correlation with benzo[a]pyrene monooxygenase activity. AB - The mutagenicity of benzo[a]pyrene and 2-aminoanthracene in the Ames test using the S-9 fraction from the liver of the Northern pike (Esox lucius) was tested. S 9 fractions were prepared both from fish injected intraperitoneally with 3 methylcholanthrene and from control animals. In addition benzo[a]pyrene monooxygenase activity was assayed in the same S-9 fractions used in the Ames test. S-9 fractions from the liver of the Northern pike were found to convert benzo[a]pyrene and 2-aminoanthracene to mutagenic metabolites. The number of revertants obtained was increased 2--4-fold in the case of 2-aminoanthracene and 3--14-fold in the case of benzo[a]pyrene by pretreatment of the pike with a single intraperitoneal injection of 3-methylcholanthrene. This injection also caused a 2--6-fold increase in the benzo[a]pyrene monooxygenase activity of the S 9 fractions used. These increases in mutagenicity and activity of the S-9 fractions used. these increases in mutagenicity and activity occur mainly during the first 4--12 days after the injection, but further small increases are observed for as long as 60 days. A strong positive correlation was found between the benzo[a]pyrene monooxygenase activity of the S-9 fractions used and their ability to give rise to revertants in the Ames test using 2-aminoanthracene or benzo[a]pyrene. This indicates that the major determining factor in the production of reactive metabolites which attack DNA in this in vitro system is the activity of the phase I cytochrome P-450 system. PMID- 6284389 TI - Effect of 2(3)-tert-butyl-4-hydroxyanisole on benzo[a]pyrene metabolism and DNA binding of benzo[a]pyrene metabolites in isolated mouse hepatocytes. AB - Benzo[a]pyrene (BP) metabolism and the conjugation and DNA-binding of BP metabolites, was studied using isolated hepatocytes from mice maintained on a diet containing 2(3)-tert-butyl-4-hydroxyanisole (BHA) (7.5 g/kg food) to discover the mechanisms involved in the anticarcinogenic effects of this antioxidant. The antioxidant feeding produced: (a) profound differences in the BP metabolite pattern, (b) no increase in the levels of either the glucuronic acid, the sulfate or the glutathione conjugates and(c) a marked decrease in the level of BP metabolites bound to intracellular DNA. Therefore, the inhibition of DNA binding observed after administration of BHA, may be due to the change in BP metabolism rather than to an increase in the conjugation of reactive metabolites. PMID- 6284390 TI - High performance liquid chromatographic assay of cefsulodin, cefotiam and cefmenoxime in serum and urine. PMID- 6284391 TI - Insulin-like activity of proteases. V. Stimulation of cyclic adenosine 3',5' monophosphate phosphodiesterase by an N-succinyl-L-alanyl-L-alanyl-L-alanine p nitroanilide-hydrolyzing protease. PMID- 6284392 TI - Synthesis of the nonatriacontapeptide corresponding to the entire amino acid sequence of ostrich adrenocorticotropic hormone. PMID- 6284393 TI - Studies on deoxynucleic acids and related compounds. IV. Syntheses of an octanucleotide containing a recognition site for restriction enzyme Eco RI and of an arabinosyladenine analog. PMID- 6284394 TI - [Our experience in salivary gland pathology of surgical importance]. PMID- 6284395 TI - High-dose cyclophosphamide with autologous marrow transplantation as initial treatment of small cell carcinoma of the bronchus. AB - Sixteen patients with untreated small cell carcinoma of the bronchus received cyclophosphamide in a total dose of 160-200 mg/kg. Autologous marrow transplantation was used to minimise the period of hypoplasia and 2 mercaptoethane sulphonate to prevent urothelial toxicity. The procedure was well tolerated, with predictable and manageable toxicity. Complete radiological and bronchoscopic response was achieved in seven patients and partial response in a further seven. High-dose cyclophosphamide may be a useful initial treatment for this disease. PMID- 6284396 TI - Treatment of vincristine-induced ileus with sincalide, a cholecystokinin analog. AB - Sincalide, a synthetic analog of cholecystokinin capable of stimulating bowel motility, has been administered to 12 patients with symptoms and signs of vincristine-induced ileus. Patients were given intravenous infusions of sincalide 0.01 microgram/kg/h over 2-24 h (mean, 8 h) for 1-12 days (mean, 5 days), usually until all evidence of ileus had resolved. Improvement was noted within 48 h of initiation of therapy in eight patients (75%), whereas marginal or no responses were observed in four patients. The mean duration of ileus was 3.6 days in responders and 7.7 days in nonresponders. Toxicity was minimal and consisted of diarrhea in two patients, in whom symptoms promptly resolved with discontinuation of the drug. Further explorations of this promising agent in the treatment of vincristine-induced ileus appears warranted. PMID- 6284397 TI - Modulation of microsome-mediated benzo[a]pyrene-metabolism by serum. AB - The effects of serum on the metabolism of benzo[a]pyrene (BP) by liver microsomes from 3-methylcholanthrene-treated rats were studied. In the presence of serum, the aryl hydrocarbon hydroxylase activities were enhanced to 160-570% of the control value. Enhancement of BP-metabolism by serum was also revealed by high performance liquid chromatography, but the relative amounts of a series of BP metabolites were not changed appreciably by addition of serum. Binding of BP metabolites to DNA was influenced by serum: adducts derived from two stereo isomeric BP-7,8-di-hydrodiol-9,10-oxides were increased, while those from 9 hydroxybenzo[a]pyrene-4,5-oxide were reduced. PMID- 6284398 TI - Anticarcinogenic and cocarcinogenic effects of benzo[e]pyrene and dibenz[a,c]anthracene on skin tumor initiation by polycyclic hydrocarbons. AB - In the present study, we have examined the effects of benzo[e]pyrene (B[e]P) and dibenz[a,c]anthracene (DB[a,c]A) on the skin tumor-initiating activities of methylated and non-methylated polycyclic aromatic hydrocarbons (PAH). B[e]P, when applied 5 min prior to initiation with seven different PAH skin carcinogens, effectively inhibited the tumor-initiating activities of 7,12 dimethylbenz[a]anthracene (DMBA) and dibenz[a,h]anthracene (DB[a,h]A) but had little or no effect on the tumor-initiating activities of 3-methyl-cholanthrene (MCA), 7-methylbenz[a]anthracene (7-MBA), 12-methylbenz[a]anthracene (12-MBA), and 5-methyl-chrysene (5-MeC). B[e]P potentiated the tumor-initiating activity of benzo[a]pyrene (B[a]P) by approximately 30%, DB[a,c]A, when applied 5 min prior to initiation, inhibited the tumor-initiating activities of DMBA, MCA, and DB[a,h]A but had little or no effect on the tumor-initiating activities of B[a]P, 7-MBA, 12-MBA, and 5-MeC. DB[a,c]A, when applied 12, 24, or 36 h prior to initiation with B[a]P, which allowed time for induction of epidermal monooxygenase enzymes, inhibited tumor initiation. The covalent binding of DMBA and B[a]P to epidermal DNA was examined under the influence of B[e]P. Doses of 20 and 200 nmol B[e]P given 5 min prior to 10 nmol [3H]DMBA reduced binding to 47 and 22%, respectively, of the control value. In contrast, doses of 200 or 2000 nmol B[e]P given 5 min prior to 200 nmol [3H]B[a]P had little or no effect on total binding. The data indicate that one cannot predict anti and cocarcinogenic effects of B[e]P and DB[a,c]A on the basis of a presence or absence of a methyl substituent. In addition, fundamental differences exist in the processing and metabolism of DMBA and B[a]P by mouse epidermal cells. PMID- 6284399 TI - Improved transformation of C3H10T1/2CL8 cells by direct- and indirect-acting carcinogens. AB - Oncogenic transformation of C3H10T1/2CL8 cells was improved by treating the cells 5 days after seeding. Benzo[a]-pyrene-induced transformation was increased 3.5 fold by this method, compared with treating the cells 1 day after seeding. N Methyl-N'-nitro-N-nitrosoguanidine, which does not transform asynchronous cultures of C3H10T1/2CL8 cells when administered 1 day after seeding, produced on average of 1 focus/dish, with 61% of the dishes exhibiting foci, when administered 5 days after seeding. Propane sultone and aflatoxin B1 also produced marked transformation responses when administered 5 days after seeding. However, 4-dimethylaminoazobenzene did not induce transformation when administered either 1 day or 5 days after seeding. With all chemicals examined, clonal cytotoxicity was reduced when they were administered 5 days after seeding. These results indicate the utility of this new procedure for the qualitative analysis of the transforming ability of chemicals. PMID- 6284400 TI - In vitro enzymatic recognition of DNA modified by O,O'-diacetyl or O-acetyl derivatives of the carcinogen 4-hydroxyaminoquinoline-1-oxide. AB - Purified DNA was modified in vitro by 3H-labelled O-acetyl or O,O'-diacetyl-4 hydroxyaminoquinoline-1-oxide (Ac4HAQO or di Ac-4HAQO). It was then subjected to the action of the single-stranded DNA specific nuclease S1 and the digested fractions were analysed. For both types of modified DNA, the release of non modified nucleotides was faster than the release of modified nucleotides. This result is at variance with that obtained with acetoxy-acetylaminofluorene modified DNA: in the latter case, the modified nucleotides were preferentially released. The results suggest that the S1 endonuclease can recognize different conformational changes in DNA, which depend on the carcinogen used. The enzymatic activity (or activities) present in Micrococcus luteus cell extracts released ethanol-soluble products from Ac-4HAQO modified DNA. PMID- 6284401 TI - Chemical interactions with herpes simplex type 2 virus: enhancement of transformation by norharman. AB - The effects of the comutagen norharman on herpes simplex virus type 2 growth transformation was studied in 3T3 cells. Pre-exposure but not post-exposure of the cells to 0.5 to 10 micrograms of norharman per ml of cell culture media resulted in a nearly three-fold enhancement in transforming activity. These results suggest that certain environmental chemicals may interact with cells creating an increased possibility of transformation by herpes viruses. PMID- 6284402 TI - Benzo[a]pyrene and other inducers of cytochrome P1-450 inhibit binding of epidermal growth factor to cell surface receptors. AB - The binding of 125I-labelled epidermal growth factor (EGF) was utilized to monitor possible cell surface effects of polycyclic aromatic hydrocarbon carcinogens. Exposure of confluent C3H 10T1/2 mouse fibroblasts to 1 muM benzo[a]pyrene led to a time-dependent decrease of EGF binding. By 24 h, EGF binding was only 5% that of control cultures. In contrast, benzo[a]pyrene-7,8 diol-9,10-oxide did not significantly alter EGF binding, indicating that the inhibition by benzo[a]pyrene was not simply due to DNA damage. A curvilinear Scatchard plot in the control cells was consistent with the presence of two classes of EGF receptors having differing affinities. Our results suggest that the major effect of benzo[a]pyrene was a reduction in receptor number rather than affinity, although other interpretations have not been excluded. Progesterone, 17 beta-estradiol, benzo[e]pyrene, cholesterol, phenobarbital, 1,1-bis-(p chlorophenyl)-2,2,2-trichloroethane, hexachlorobenzene or pregnenolone-16 alpha carbonitrile, did not inhibit EGF binding. On the other hand, several known inducers of P1-450 were very effective inhibitors of EGF binding. These included: dimethylbenz[a]anthracene, 3-methylcholanthrene, benzo[a]pyrene, benz[a]anthracene, beta-naphthoflavone and alpha-naphthoflavone. We postulate that the binding of certain polycyclic aromatic hydrocarbons to the Ah receptor may induce not only specific drug metabolizing enzymes but also inhibition of EGF binding, and possible other cell effects. Further studies are required to verify this hypothesis. PMID- 6284403 TI - Effect of nonoxynol-9, a detergent with spermicidal activity, on malignant transformation in vitro. PMID- 6284404 TI - Pulmonary cellular dysfunction in endotoxin shock: metabolic and transport derangements. AB - Lung cell metabolic and ion transport alterations in endotoxin shock have been studied to elucidate mechanisms of endotoxin-induced pulmonary dysfunction. Glucose oxidation to lactate as well as to CO2 is significantly increased with a decrease in the lung ATP contents. Active Na+ transport is impaired, indicating an increase in lung intracellular fluid. The altered glucose metabolism and active ion transport could potentially contribute to lung edemagenesis in endotoxicosis. PMID- 6284405 TI - Transient depolarization and spontaneous voltage fluctuations in isolated single cells from guinea pig ventricles. Calcium-mediated membrane potential fluctuations. AB - Under the influence of cardiotonic steroids, single ventricular cells exhibit transient depolarization after a train of driven action potentials or, in voltage clamp experiments, transient inward current after a depolarizing clamp pulse. Transient depolarization or transient inward current was abolished by an intracellular injection of ethyleneglycol-bis (beta-aminoethyl ether)-N,N' tetraacetic acid (EGTA) or by superfusion of 5 mM caffeine. Transient depolarization was elicited even in the control Tyrode's solution by an intracellular injection of CaCl2 or augmented by an injection of adenosine 3',5' cyclic monophosphoric acid (cAMP). Along with transient depolarization or transient inward current, digitalis intoxication promoted spontaneous oscillatory fluctuations in membrane potential or in membrane current. Their power spectra showed peaks at frequencies ranging from 2 to 7 Hz, which coincided well with the frequency of repetitive transient depolarization or transient inward current. The fluctuations were eliminated by intracellular injections of EGTA and decreased in amplitude by 5 mM caffeine with a shift toward higher frequencies. Depolarization of the membrane caused a shift of the spectrum peak toward higher frequencies. These results suggest that an oscillatory release of Ca from intracellular storage sites is the common basis underlying both the transient events (depolarization or inward current) and the spontaneous miniature fluctuations in membrane potential or current. PMID- 6284406 TI - Identification of alpha 1-adrenergic receptors in cultured rat myocardial cells with a new iodinated alpha 1-adrenergic antagonist, [125I]IBE 2254. PMID- 6284407 TI - Evaluation of noninvasive tests of cardiac damage in suspected cardiac contusion. AB - Nonpenetrating trauma to the chest can result in cardiac damage that may be overlooked because of associated injuries and the lack of obvious thoracic injury. The clinical diagnosis of important cardiac damage in this setting is difficult. We evaluated noninvasive tests for detecting myocardial damage in 100 patients with severe, nonpenetrating chest trauma. The noninvasive tests included serial ECG, serial total CPK and CPK-MB enzymes, continuous Holter monitor recording to detect dysrhythmia, and technetium-99m pyrophosphate scintigraphy. Peak CPK-MB elevations occurred in 72 patients. ECG abnormalities were noted in 70 patients, and 27 patients had Lown grade 3 or greater dysrhythmias. Fifteen patients died and all had autopsies. The noninvasive abnormalities were nonspecific and did not reflect myocardial contusion that led to clinically important cardiac complications. PMID- 6284409 TI - A sensitive enzyme immunoassay specific for human chorionic gonadotrophin. PMID- 6284408 TI - Functional activity of intrinsic factor measured by using solubilized receptor protein. AB - The traditional radioimmunoassay for gastric intrinsic factor, in which this protein is measured on the basis of immunoreactivity rather than function, is of no value for identifying intrinsic factor that binds cobalamin but does not bind to the ileal receptor site, or for detecting animal intrinsic factor, which does not cross react with human intrinsic factor. Accordingly, we have applied a radioassay for the intrinsic factor receptor protein to measure the functional activity of intrinsic factor in gastric juice. The receptor protein reagent was partly purified from guniea pig ilea and its interaction with intrinsic factor- CN[57Co]-cobalamin was determined by precipitation with sodium sulfate at a final concentration of 150 g/L. Results of this assay were comparable with results obtained for intrinsic factor by radioimmunoassay. The receptor protein did not bind immunoreactive intrinsic factor that was functionally abnormal. This functional radioassay for intrinsic factor is not species specific and will be of value when specific antiserum to intrinsic factor is not available and when cobalamin malabsorption is to be evaluated in patients who are secreting normal amounts of immunoreactive intrinsic factor. PMID- 6284410 TI - Demonstration of circulating 1,24,25-trihydroxyvitamin D3 in man by radioimmunoassay. AB - 1,24,25-trihydroxyvitamin D3 has been detected in human serum using a sensitive radioimmunoassay. Tritiated 1,24,25-trihydroxyvitamin D3 was synthesized biologically and used as tracer to monitor the recovery of endogenous metabolite during isolation from serum. Circulating 1,24,25(OH)3D3 in normal subjects ranged from 9.3 to 18.5 pmol/l but was not detectable (less than 2.3 pmol/l) in serum from nephrectomized subjects. The trihydroxymetabolite was elevated in three out of four vitamin D deficient subjects who were being treated with vitamin D3. PMID- 6284411 TI - Studies on the mechanism of the enhancement of the antihypertensive activity of captopril by a diuretic in spontaneously hypertensive rats. AB - Captopril (30 mg/kg/day orally for two days) in spontaneously hypertensive rats (SHR) inhibited serum angiotensin converting enzyme (ACE) activity 92.3%; increased plasma renin activity (PRA) 18-fold and reduced mean arterial blood pressure (MABP) 19 mm Hg. Hydrochlorothiazide (HCTZ) (100 mg/kg-day 1; 10 mg/kg day 2, orally) increased PRA 3-fold but did not affect serum ACE or MABP. HCTZ plus captopril inhibited serum ACE 95.2%; increased PRA 38-fold and reduced MABP 47.5 mm Hg. Captopril or HCTZ plus captopril did not alter the responses of isolated aortic strips to norepinephrine (NE), serotonin, angiotensin II (AII) or isoproterenol. Pressor responses of conscious SHR to AII and NE were unaltered by captopril or HCTZ plus captopril although the bradykinin-induced depressor responses were significantly but equally potentiated. These results suggest that the potentiating effect of HCTZ is due to some mechanism that shifts the animal's blood pressure maintenance system to a renin-dependent state and is not due to changes in vascular reactivity. PMID- 6284412 TI - Impairment of neurally-mediated vasoconstriction in DOCA-salt hypertensive dogs. AB - Several biochemical studies indicate that hypertension produced by administration of deoxycorticosterone acetate (DOCA) plus saline as the drinking fluid is accompanied by significant increases in catecholamine turnover rates and a decrease in catecholamine content of peripheral tissues. In the present study the functional significance of such biochemical changes was assessed by evaluating vasoconstrictor responsiveness to neurogenic and humoral interventions in DOCA salt hypertensive dogs. Administration of DOCA, 5 mg/kg subcutaneously twice a week for four weeks to unilaterally nephrectomized dogs given drinking fluid containing 1% sodium chloride produced a significant increase in blood pressure within one week. At the end of 4 week treatment period, the animals were anesthetized with pentobarbital (35 mg/kg) and sympathetic neurotransmission to the hindlimb and kidney was evaluated. Femoral as well as renal vasoconstrictor responses to sympathetic nerve stimulation were significantly inhibited in the hypertensive group in comparison with control animals. Vasoconstriction elicited by exogenous norepinephrine in both these vasculatures was similar in control and hypertensive groups. Femoral vasodilator responses to adenosine, bradykinin and PGE2 were significantly attenuated in the hypertensive group. While indomethacin (10 mg/kg) did not significantly affect renal vasoconstrictor responses to renal nerve stimulation in both the groups of animals, it caused a significant increase in the blood pressure of hypertensive group but not of control group. These results do not support the hypothesis of an increase in neurogenic function contributing to DOCA-salt hypertension. It is suggested that a reduction in sympathetic neurogenic influence may serve to protect against the rise in blood pressure during DOCA-salt administration. PMID- 6284413 TI - [Oxytocin receptor--its effects on inducing uterine contraction (author's transl)]. PMID- 6284414 TI - [Radioimmunoassay for gonadoliberin with active florisil (author's transl)]. PMID- 6284415 TI - Enzymatic disorders and hypertension. PMID- 6284416 TI - Cushing's syndrome and exogenous glucocorticoid hypertension. PMID- 6284417 TI - The kinins and prostaglandins in hypertension. PMID- 6284418 TI - Phaeochromocytoma. PMID- 6284419 TI - Hypothalamic--pituitary function in the fetus and infant. PMID- 6284421 TI - Hereditary pyrophosphate arthropathy (familial articular chondrocalcinosis) in Sweden. AB - Genealogical links between three Swedish families with hereditary pyrophosphate arthropathy were found in the 18th century, indicating a possible founder effect, similar to earlier findings in Slovakia, France and Chile. However, no connection between the Swedish and other European families with the disease has so far been found. In accordance with other reported familial aggregations of pyrophosphate arthropathy, the transmission of the disease in the Swedish families appeared to be autosomal dominant with incomplete penetrance and variable expressivity. Severe symptoms related to homozygotic cases reported in some other families were not found in Sweden. PMID- 6284420 TI - Pubertal development: normal, precocious and delayed. AB - The present concepts on the neuroendocrine mechanisms which trigger pubertal development and modulate the progression towards sexual maturity have been reviewed. Essentially, puberty is presented as a continuum, the programming of which is initiated prenatally and which ends in adult life when all hormonal secretions become autoregulated. This continuum is dependent on a delicate equilibrium between CNS neurohormones (GnRH), neurotransmitters (biogenic amines), pituitary gonadotrophin (FSH, LH) secretion and the end-organ response (testis or ovary) through the activation of specific membrane receptors. The gonadal sex steroids (T, OE2) will activate specific cytoplasmic and nuclear receptors of target tissues and exert their biological action. Initially, the activity of the HPGA is manifested by nocturnal LH peaks, followed by increased gonadal secretion of T or OE2. Extremely sensitive to negative feedback by circulating androgen and/or oestrogen in prepuberty, an hypothalamic regulatory system called the gonadostat increases its threshold of sensitivity and eventually becomes autoregulated at a higher feedback level. Progressively, the hypothalamus becomes sensitive to positive feedback action of gonadal hormones, this phenomenon being important for the onset of ovulation. It is likely also that adrenal androgens play a permissive and supportive role in the onset and progression of pubertal development. Finally, full maturity is reached, with final adult height through fusion of the epiphysis, and fertility is achieved. The clinical manifestations of each developmental stage of puberty are described and abnormalities of sexual development reviewed. While over 90 per cent of cases of precocious pubertal development are idiopathic in girls, a space-occupying lesion in the hypothalamic-pituitary region is frequent in boys. Dissociated pubertal signs (premature adrenarche, pubarche, thelarche, menarche) are discussed, together with diagnosis and treatment of precocious puberty, whether it is idiopathic or occurring independently of the activation of the HPGA. In addition to delay of puberty on a constitutional basis, or related to chronic endocrine or non-endocrine diseases, the main clinical entities with gonadal insufficiency, primary (hypergonadotrophic) or secondary (hypogonadotrophic), are reviewed in boys and girls and their investigation and treatment discussed. PMID- 6284422 TI - Selective immunosuppressive effects of measles virus infection. AB - The antibody response to diphtheria toxoid by cultured tonsil cells was suppressed by measles virus, which productively infected a small percentage of both T and B lymphocytes. However, infection of cultures spontaneously secreting antibody and immunoglobulin (with Herpes simplex virus) or measles virus did not result in immunosuppression. Immunosuppression by measles virus may thus be attributed to a selective effect on the inductive phase of the response and suggest that infection of immunoglobulin-producing B lymphocytes does not alter the ability of these cells to secrete antibody. PMID- 6284423 TI - Cytomegalovirus infection in infancy: virological and immunological studies. AB - Immunological and virological studies on 18 infants with cytomegalovirus (CMV) infection were performed. Eleven of these infants were studied on multiple occasions over a period of 1 year. The patients were divided into three clinical groups based on the probable time of infection and the resulting variation in clinical presentation. General parameters of cell-mediated immunity as determined by E-rosette formation and lymphocyte proliferative responses to mitogens and antigens were found to be normal. Quantitation of CMV excretion in urine, CMV specific immunofluorescent (IF) and complement-fixing (CF) antibody titres and CMV-specific cell-mediated immune responses were done on all patients at approximately monthly intervals. Throughout the study period all patients continued to excrete CMV despite the presence of high antibody titres to the virus. CMV-specific lymphocyte proliferative responses were absent or diminished in 15 of the 18 patients. The immunological and virological status of all patients was similar regardless of the clinical manifestation of infection. PMID- 6284424 TI - Further comparison of the effects of physostigmine and neostigmine on frog neuromuscular transmission. AB - 1. The effects of physostigmine and neostigmine were compared at frog neuromuscular junctions using intracellular microelectrodes. 2. Both drugs increased the amplitudes of miniature endplate potentials (MEPP) in a dose dependent manner because of their ability to competitively inhibit cholinesterase activity. 3. In addition, physostigmine (1.0-20.0 mumol/l) decreased by approximately 20% the quantal content (determined by two methods), indicating a decrease in the amount of transmitter released upon nerve stimulation, whereas neostigmine did not alter quantal content. 4. Because the endplate potential (EPP) amplitude is a composite of pre- and post-junctional effects, the dose response curve for neostigmine on EPP amplitude approximately paralleled that obtained on MEPP amplitude, whereas the effect of physostigmine on EPP amplitude was depressed because of its action to decrease transmitter release. 5. Upon washout of neostigmine, the increase in EPP amplitude reversed more rapidly than effects on MEPP amplitude, a difference dependent on the magnitude of quantal content and explained by an action on the postjunctional membrane. Similar results were obtained with physostigmine. 6. Neither drug significantly affected MEPP frequency although small decreases were sometimes observed. 7. These results suggest that at the frog neuromuscular junction the effects of neostigmine are explained primarily by its inhibition of cholinesterase, whereas physostigmine are explained primarily by its inhibition of cholinesterase, whereas physostigmine has an additional action on nerve terminals of decreasing nerve stimulated release of acetylcholine. PMID- 6284425 TI - Characterization and localization of [3H]-clonidine binding in membranes prepared from guinea-pig spleen. AB - 1. [3H]-Clonidine binds to membranes prepared from guinea-pig spleen with high affinity. 2. Kinetic experiments indicated that [3H]-clonidine associates rapidly to the binding site and that the binding is reversible. A study of the dissociation of [3H]-clonidine from splenic membranes revealed two components. The slowly dissociating component corresponded to a high affinity process (Kd = 2.1 nmol/l) in good agreement to that obtained by saturation analysis. 3. Over the concentration range used, saturation experiments revealed only a single population of sites with a dissociation constant (Kd) of 2.4 nmol/l and a density of 5.1 pmol/g wet weight tissue. 4. Examination of the relatively potency of a series of alpha-adrenoceptor agonists and antagonists indicates that [3H] clonidine binding is to alpha 2-adrenoceptors. 5. High levels of binding were obtained to lymphocytes prepared from guinea-pig spleen and to membranes from the splenic capsule. Pretreatment of animals with 6-hydroxydopamine produced changes in apparent affinity of binding with little change in the number of receptor sites. 6. It is concluded that [3H]-clonidine labels is a site resembling the alpha 2-adrenoceptor in guinea-pig spleen. Few if any of these sites are located prejunctionally and a significant fraction are associated with lymphocytes. PMID- 6284426 TI - Primitive neuroectodermal tumor arising in the ulnar nerve. A case report. AB - Malignant peripheral nerve tumors are rare. A neuroectodermal tumor arose in a 59 year-old man from the ulnar nerve. Typically, the tumor reproduces some of the appearances of cells of the primitive central nervous system. The possibility that a primary tumor of the sympathetic nervous system, especially of the adrenal glands, should be considered in the differential diagnosis. The tumor may arise from a major peripheral nerve and show features of growth patterns of the embryonic central nervous system. Blood stream spread is not uncommon; awareness of this proclivity makes early diagnosis mandatory. PMID- 6284427 TI - Response to parathyroid and calcitonin of human giant cell tumor of bone cultured in patients' serum. PMID- 6284428 TI - [Ramsay Hunt syndrome-like symptom complex associated with isolated ACTH deficiency; a case report (author's transl)]. PMID- 6284429 TI - Renal scanning: pitfalls in the diagnosis of mass lesions. PMID- 6284430 TI - Lymphocyte beta-adrenergic receptors are not altered in hyperthyroidism. AB - Binding of (-)3H]-dihydroalprenolol (3H-DHA) to lymphocytes from five thyrotoxic patients and five age- and sex-matched contents were examined to ascertain whether beta-adrenergic receptor number or binding affinity were altered in thyrotoxicosis. Whereas an increase in beta-adrenoceptor density has been reported in triiodothyronine-induced hyperthyroidism, we did not find changes in beta-adrenergic receptor density or binding affinity. In one patient studied sequentially, we did not find alteration in 3H-DHA bindings before or after the subject was rendered euthyroid. We conclude that lymphocyte beta-adrenergic receptor density and affinity are not altered in spontaneous hyperthyroidism. PMID- 6284432 TI - Vitamin D3 and calcium absorption. PMID- 6284431 TI - Effects of increasing urinary pH on cigarette smoking. AB - We evaluated the effects of increasing urinary pH on sensitive automated measures of cigarette smoking behavior. Urine was alkalized by oral doses of 50 or 75 mg/kg sodium bicarbonate. Measurements of cigarette smoking behavior included the number of cigarettes smoked and automated measures of puff frequency and duration. The sodium bicarbonate resulted in little or no change in the number of cigarettes smoked, but in five of the seven subjects frequency and duration of cigarette puffs after sodium bicarbonate decreased 10% to 15%. The small decreases in cigarette smoking brought about by sodium bicarbonate cannot be expected to have significant overall effects. PMID- 6284433 TI - Increased angiotensin-converting enzyme activity of aorta in two-kidney, one-clip Goldblatt hypertension in rats. PMID- 6284434 TI - [Clinical study of the secondary toxicity, effective responses and survival time in the treatment of primary non-microcytic (NSCLC) carcinoma of the lung with various polychemotherapy regimens]. PMID- 6284435 TI - Pathology of carcinoma of the lung. Changing patterns. PMID- 6284436 TI - Production of biochemical marker substances by bronchogenic carcinomas. PMID- 6284437 TI - Surgical therapy for carcinoma of the lung. AB - Surgical resection in patients with carcinoma of the lung remains the one therapeutic modality that offers the greatest potential for long-term survival. However, it is apparent that the surgical treatment of carcinoma of the lung must be applied selectively. The selection of the appropriate patients is determined by the extent of the disease process, the cell type of the tumor, and the physical status of the patient. The operative procedure chosen in a given patient may be either conservative or extended in nature as deemed necessary to eradicate the local disease process. The morbidity and mortality must be commensurate with the potential salvage that may be expected. The latter is primarily determined by the extent of the disease process at the time of operation. PMID- 6284438 TI - Small cell cancer of the lung. PMID- 6284439 TI - Management of the patient with multiple sclerosis. PMID- 6284440 TI - Endobronchial involvement in metastatic breast carcinoma. PMID- 6284441 TI - Hepatoma presenting as a cerebellar metastasis. PMID- 6284442 TI - Corneal epithelial healing rates after topical nucleotides. AB - Discrete 28mm2 epithelial lesions in paired rabbit corneas were treated four times daily with either vehicle alone or 10(-2)M or 10(-3)M Db cAMP in the presence or absence of either theophylline (10(-2)M) or epinephrine (5.5 x 10( 3)M). Cyclic GMP was also used either alone or with pilocarpine (1%). The lesion size was determined photographically at various intervals over 48 hours. No treatment regimen at the concentrations of cyclic nucleotides used had any influence on the rate of epithelial regrowth. PMID- 6284443 TI - Structure and assembly of alphaviruses. PMID- 6284444 TI - Cyclic nucleotide metabolism in the vertebrate retina. PMID- 6284445 TI - Severe hepatitis associated with ketoconazole therapy for chronic mucocutaneous candidiasis. PMID- 6284446 TI - The influence of corticosteroids and theophylline on cerebral function. A review. AB - Evidence indicating that corticosteroid therapy may adversely affect attention and memory in asthmatic subjects is reviewed. The potential impact of corticosteroid dosage levels (high versus low) and treatment regimens (daily versus alternate day) on attention and memory is discussed. Findings indicate that the suppression of ACTH by corticosteroids may have different effects on attention and memory as a function of the sex of the asthmatic patient. The need to establish what the effects of theophylline-induced reductions of cerebral blood flow and cerebral oxygen tension are on higher cognitive processes are outlined. The importance of establishing whether or not medications commonly used for the treatment of asthma create deficits of memory or attention that interfere with medical compliance is emphasized. PMID- 6284447 TI - Virus isolation and identification in cervical cancer patients. PMID- 6284448 TI - Gray-scale ultrasonography, an integral part of diagnostic radiology. PMID- 6284449 TI - Diagnostic problems of insulinoma. PMID- 6284450 TI - [Clinical observation on 90 cases of malignant trophoblastic neoplasms treated with gengxinmycin (author's transl)]. PMID- 6284451 TI - The organization of macronuclear rDNA molecules of four hypotrichous ciliated protozoans. AB - We have compared the structure of macronuclear DNA molecules that contain rRNA genes of four hypotricous ciliates, Stylonychia pustulata, Euplotes aediculatus, Oxytricha fallax and Oxytricha nova. The macronuclear rDNA, like all macronuclear DNA in hypotrichs, exists as achromosomal molecules of approximately single-gene size. The rDNA molecules have been cloned intact as recombinant plasmids and analyzed by restriction mapping and Southern hybridization. The sites of restriction enzymes BamHI, EcoRI, HindIII, PstI, PvuII and XhoI have similar but not identical patterns in Stylonychia and the two Oxytricha rDNAs. The restriction pattern of Euplotes rDNA is unlike those of the other three, with only one site of seventeen in the same position. Despite this divergence in nucleotide sequence, the overall structure of the rDNA molecules in the four hypotrichs is constant. The size of all the rDNA molecules is the same, 7.49 kb. Also, the positions of the regions coding for 19S and 25S rRNA are alike. The 25S coding region is at the 5' end of the DNA template strand (3' end of the RNA transcript), within 500 base pairs of the terminus of the DNA molecule. The 19S coding region is adjacent to the 25S region with less than 500 base pairs of spacer lying between the two genes. The largest non-coding sequence is at the 3' end of the DNA molecule adjoining the 19S RNA gene. The 3' non-coding regions show greater sequence divergence among the different rDNAs than do the coding regions. The similarity in size and organization of these molecules and the variability in the restriction patterns suggest that the gene structure is under tighter evolutionary constraint than is the primary nucleotide sequence. PMID- 6284454 TI - [A preliminary study of concentration of cyclic AMP in the cerebrospinal fluid of neurosurgical patients (author's transl)]. PMID- 6284453 TI - [Hormones and neoplasms (author's transl)]. PMID- 6284452 TI - Elimination of DNA sequences during macronuclear differentiation in Tetrahymena thermophila, as detected by in situ hybridization. AB - Previous studies have indicated that certain sequences in the micronuclear genome are absent from the somatic macronucleus of Tetrahymena (Yao and Gorovsky, 1974; Yao and Gall, 1979; Yao, submitted). The present study used in situ hybridization to follow the elimination process during the formation of the new macronucleus. Micronuclear-specific DNA cloned in recombinant plasmids was labelled with 3H and hybridized to cytological preparations of T. thermophila at various stages of conjugation. Despite a smaller size and lower DNA content, the micronucleus has more hybridization than the mature macronucleus. Hybridization initially increased in the anlage (newly developing macronucleus) to reach a maximal level right after the old macronuclei has disappeared. The hybridization in the anlage than decreased to a significant extent prior to the first cell division. The results suggest that the micronuclear-specific sequence is first replicated a few rounds before it is eliminated from the anlage, and the elimination process occurs without nuclear division. PMID- 6284455 TI - [Value of selective angiography in diagnosis of primary hepatocellular carcinoma (author's transl)]. PMID- 6284456 TI - [Bronchiolo-alveolar carcinoma: a study of 37 cases (author's transl)]. PMID- 6284458 TI - Primary linitis plastica of the rectum: report of a case. PMID- 6284457 TI - High-fiber diet reduces bleeding and pain in patients with hemorrhoids: a double blind trial of Vi-Siblin. PMID- 6284459 TI - Ranitidine in duodenal ulcer: incidence of healing and effect of smoking. AB - The effect of ranitidine, a new H2-receptor antagonist, on the healing of duodenal ulcer has been assessed in a double-blind study. Fifty patients with endoscopically proven duodenal ulcer were randomly assigned to ranitidine 150 mg twice daily or placebo for 4 weeks. Endoscopic examination at this time showed that 20 of 25 patients (80%) on ranitidine healed compared to 4 of 25 patients (16%) on placebo (P less than 0.01). Smoking adversely affected the incidence of healing: 70% of nonsmokers healed compared to 30% of smokers (P less than 0.03). There were no side effects noted on ranitidine. Review at 6 months after cessation of therapy showed relapse of duodenal ulcer in 10 of 20 patients (50%) healed with ranitidine and 1 of 4 patients (20%) healed with placebo. Thus, ranitidine (300 mg/day) produces similar healing rates to those reported for cimetidine (1000 mg/day); also like cimetidine, the incidence of healing on ranitidine is adversely influenced by smoking and the relapse rate on cessation of therapy is high. PMID- 6284461 TI - Nesidioblastosis associated with insulin-mediated hypoglycemia in an adult. AB - Nesidioblastosis, the process of differentiation of pancreatic islets from ductular epithelium, is a well-described cause of insulin-mediated hypoglycemia in neonates and infants, but not in adults. A 58-yr-old woman with characteristic clinical features of fasting hypoglycemia had inappropriately elevated plasma immunoreactive insulin levels during symptomatic episodes of fasting hypoglycemia. Angiography, palpation at laparotomy, and resection of the distal three-quarters of the pancreas provided no evidence of a tumor. Pathologic examination of the resected pancreas revealed the findings of nesidioblastosis, i.e., budding of islets from the wall of ductules, and also increased number and size of islets and abnormal shape and location of islets. An entire spectrum of islet cell abnormalities including nesidioblastosis can cause insulin-mediated hypoglycemia in adults, as it does in neonates and infants. PMID- 6284460 TI - Gastrointestinal effects of long-term colchicine therapy in patients with recurrent polyserositis (familial mediterranean fever). AB - Twelve patients with recurrent polyserositis (RP, familial Mediterranean fever) on colchicine prophylaxis (1.0-2.0 mg daily) for three years or more were evaluated for the presence of gastrointestinal effects possibly attributable to the drug. Two patients had bulky stools, two others had transient diarrhea, and one had heartburn. Serum vitamin B12, calcium, and carotene levels were normal in all cases, and D-xylose absorption was normal in 11 of the 12. Three patients had mild steatorrhea (7.5, 7.9, and 9.9 g daily). Jejunal biopsies from these and a fourth patient with bulky stools but normal fecal fat excretion showed no abnormal histological changes. However, (Na + K)-ATPase activity was significantly decreased in all four cases. Colchicine had to be discontinued in only one of the 12 cases. It is concluded that mild steatorrhea and enzyme inhibition may occur in patients on long-term colchicine prophylaxis and that careful periodic observations for this and other adverse effects is imperative in such patients. PMID- 6284463 TI - [Ultrastructure and histogenesis of bronchiolo-alveolar carcinoma (author's transl)]. PMID- 6284462 TI - Endorphins and enkephalins. PMID- 6284464 TI - [Anti-complement immunoperoxidase technique in the determination of EBNA antibody level in patients with nasopharyngeal carcinoma (author's transl)]. PMID- 6284465 TI - [Comparative study of serum HSV-2 antibody by immuno-enzyme slide method in high and low incidence areas of cancer of the uterine cervix (author's transl)]. PMID- 6284467 TI - Papillary carcinoma arising from ectopic breast tissue in an axillary lymph node. AB - A cystic carcinoma was located in an axillary lymph node of a 90-year-old woman. The lymph node also contained cytologically benign mammary ducts and a few small glands that had epithelium cytologically intermediate between the normal epithelium and the neoplastic cells. In a subsequent mastectomy specimen, a small intraductal papillary carcinoma was found, differing cytologically and architectonically from the nodal carcinoma. The nodal carcinoma is considered primary because of its morphologic differences from the mammary lesion, the in situ nature of the mammary tumor, and the transition of benign to atypical epithelium in the nodal inclusions. The occurrence of two tumors is viewed as a field effect. PMID- 6284466 TI - Arias-Stella reaction with prominent nuclear pseudoinclusions simulating herpetic endometritis. AB - We studied a pronounced Arias-Stella reaction in an incomplete abortion curettage specimen; the gestational age was 8 weeks by size. The majority of cells in this reaction showed ground glass, eosinophilic nuclear "inclusions" simulating herpetic Cowdry type B nuclear inclusions. This was coupled with a history of herpetic stomatitis of the patient and her husband 2 weeks prior to the curettage, raising the question of viremia and transplacental herpetic endometrial infection. Immunoperoxidase staining using the PAP technique directed against Herpes simplex virus types 1 and 2 was negative. Ultrastructural analysis elucidated the true nature of the nuclear "inclusions" as pseudonuclear inclusions consisting of invaginations of cytoplasm within the nuclei. Viral particles were not found. We are unaware of any similar report, and present this case to suggest the possible differential diagnosis between herpetic endometritis with associated cytologic atypia and the Arias-Stella reaction. PMID- 6284468 TI - Utility of diagnostic imaging in the staging of gestational trophoblastic disease. AB - Gestational trophoblastic disease (GTD) is a group of tumors with a cure rate of 90-100% with appropriate treatment. The patients with a poor prognosis have metastatic disease involving structures other than the pelvic organs or the lungs. The medical records of 70 patients with histologically proven GTD were reviewed to determine the usefulness of diagnostic imaging modalities in the staging and follow-up of GTD. The level of chorionic gonadotropin (hCG) is more sensitive in determining the persistence of disease or its response to therapy, but is of little use in evaluating the site of metastases. Diagnostic imaging modalities are most useful in determining the presence and sites of metastases so that appropriate treatment is instituted. PMID- 6284469 TI - [Adrenaline oxidation by tumor nuclear membranes mediated by superoxide radicals]. PMID- 6284471 TI - Mathematical modeling of ionic processes in human skeletal muscle fibres. PMID- 6284472 TI - Electrophysiological studies in myotonic distrophy. PMID- 6284473 TI - Diagnostic yield of single fiber electromyography and other electrophysiological techniques in myasthenia gravis. II. Jitter and motor unit fiber density studies. Clinical remission and thymectomy evaluation. PMID- 6284470 TI - Trifluridine: a review of its antiviral activity and therapeutic use in the topical treatment of viral eye infections. AB - Trifluridine (trifluorothymidine) is an antiviral agent for topical use in the eye, and is structurally related to idoxuridine. In vitro studies have shown that it effectively inhibits the replication of herpes simplex virus type 1, which causes primary keratoconjunctivitis and recurrent epithelial keratitis in man. In masked comparative studies, predominantly in patients with dendritic ulcers, trifluridine 1% solution was effective in over 90% of patients; in such studies it was comparable with vidarabine in treating dendritic ulcers, and was at least as effective as, and in some studies more effective than, idoxuridine. The drug was also effective in treating a small number of patients with geographic ulcers (sometimes associated with the usage of topical corticosteroids), and this could be an important advantage if confirmed in further well-designed studies. However, experience at present is too limited to reliably determine the usual response rate in this difficult therapeutic area. In open studies the drug proved to be particularly useful in treating ulcers previously unresponsive to idoxuridine or vidarabine, and in treating patients intolerant of idoxuridine, with a high success rate and minimal side effects being reported. The role of trifluridine in treating deep stromal disease, uveitis, or adenovirus kerato-conjunctivitis has not been established. The drug is well tolerated and cross-hypersensitivity and cross-toxicity between trifluridine, idoxuridine and vidarabine are rare. Thus, trifluridine is an effective alternative to the drugs available for treating herpetic keratitis, and seems especially useful in 'difficult' cases. PMID- 6284474 TI - [Effects of clonidine on opiate withdrawal symptoms. Results - biochemical mechanisms (author's transl)]. AB - Clonidine was administered to nineteen patients in an inpatient setting after abrupt discontinuation of chronic opiate addiction (morphine, heroin, dextromoramide). Clonidine produces a decrease sometimes very rapid in opiate withdrawal signs but does not suppress the whole affects associated with. These data support the hypothesis that clonidine has antiwithdrawal effect by replacing opiate-mediated inhibition with alpha 2 mediated inhibition of brain noradrenergic activity. PMID- 6284475 TI - Effects of (-)-trans- delta 9-tetrahydrocannabinol on serum prolactin in the pseudopregnant rat. AB - Groups of pseudopregnant rats were injected intravenously with (-)-trans- delta 9 tetrahydrocannabinol (THC) to determine its effects on serum prolactin (PRL) and the maintenance of pseudopregnancy. A single injection of 4 mg THC/kg BW at 2400 h on the first day of leukocytic vaginal smears of pseudopregnancy (D-1) delayed the ensuing nocturnal PRL surge for approximately one hour. When THC (1.0 mg/kg BW) was administered hourly from 2400 h on D-1 through 1700 h on D-2, the nocturnal surge was blocked and serum PRL levels were suppressed until 0600 h on D-2, but not thereafter. Neither treatment altered the duration of pseudopregnancy. These results indicate that the nocturnal surge secretion of PRL during early pseudopregnancy in the rat is sensitive to THC suppression, but that this suppression is not adequate to influence the duration of pseudopregnancy. The mechanism through which THC exerts this action remains unknown. PMID- 6284477 TI - Physiological concentrations of dopamine decrease adenosine 3',5'-monophosphate levels in cultured rat anterior pituitary cells and enriched populations of lactotrophs: evidence for a causal relationship to inhibition of prolactin release. PMID- 6284478 TI - Effects of a prolactin- and adrenocorticotropin-secreting tumor on gonadotropin levels and accessory sex organ weights in adult male rats: a possible role of the adrenals. AB - The presence of the transplantable PRL- and ACTH-secreting tumor 7315a in intact male rats resulted in very high serum PRL levels, decreased levels of LH and FSH, and reduced weights of testes and accessory sex organs. In gonadectomized rats bearing a small testosterone-filled capsule, the tumor inhibited the postcastration rise in gonadotropins and reduced the weights of the prostate and seminal vesicles. However, after adrenalectomy of intact or gonadectomized male rats bearing a small testosterone-filled capsule, inhibitory effects of the tumor on serum gonadotropin levels or on reproductive organ weights were totally absent. These results show that in adrenalectomized male rats hyperprolactinemia in itself doses not affect gonadotropin secretion and the androgenic action of testosterone. Rather, the tumor might exert a gonadotropin inhibitory action through elevated levels of PRL combined with progesterone, which is secreted by the ACTH-stimulated adrenal. PMID- 6284476 TI - Stimulation by calcium and other cations of the cholesterol binding to steroid free cytochrome P-450scc purified from bovine adrenocortical mitochondria: the implication of ACTH-mediated calcium homeostasis on the cholesterol availability. AB - By utilizing purified cytochrome P-450scc from bovine adrenocortical mitochondria and cholesterol-containing dioleoylglycerophosphocholine liposomes, we have demonstrated a dramatic stimulation (2-3 fold) of cholesterol binding to the steroid-free cytochrome by Ca++. We theorize that ACTH modulates the increase of intracellular Ca++ concentration resulting in the increase of the cholesterol availability to the mitochondrial cytochrome. PMID- 6284479 TI - Characterization of gamma-melanotropin-like immunoreactivity and its secretion in an adrenocorticotropin-producing mouse pituitary tumor cell line. AB - gamma MSH, a putative hormone in the N-terminal region of the ACTH/beta-endorphin (beta-EP) precursor protein, was studied by RIA with an antiserum against gamma 3MSH in ACTH-producing mouse pituitary tumor cells, AtT-20/D16v. Serial dilution of the culture medium or the cell extract gave parallel lines to the standard curve in the RIA for gamma MSH. Rat median eminence extracts enhanced the release of gamma MSH-like immunoreactivity (gamma MSH-LI) concomitant with ACTH-like immunoreactivity (ACTH-LI) and beta-EP-like immunoreactivity (beta-EP-LI). Dexamethasone suppressed the release of gamma MSH-LI as well as ACTH-LI and beta EP-LI. Gel exclusion chromatography of the culture medium and the cell extract has revealed that gamma MSH-LI consists of two peaks; one eluted near the elution position of beta-lipotropin and the other near the elution position of beta-EP. There was no peak corresponding to the elution position of synthetic gamma 3MSH. However, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) has demonstrated that gamma MSH-LI migrated at five positions with molecular weights of 31K, 21-23K, 16-17K, 13-14K, and 3.8K, respectively. The 31K gamma MSH coincided with the migration position of 31K ACTH of 31K beta-EP, and 21-23K gamma MSH coincided with the position of 21-23K ACTH on SDS-PAGE. The 16-17K gamma MSH coincided with the mouse 16K fragment (reported by Eipper and Mains) of ACTH-beta-lipotropin precursor protein in the migration in SDS-PAGE and in immunoreactivity to anti-gamma MSH antiserum. [3H]Glucosamine was incorporated into 16K, 13K, and 3.8K gamma MSH. These results suggest that AtT-20/D16v cells produce gamma MSH-LIs with molecular weights of 31K, 21-23K, 16-17K, 13-14K, and 3.8K, and they are secreted concomitantly with ACTH-LI and beta-EP-LI. PMID- 6284480 TI - Mechanism of induction of delta 5-3 beta-hydroxysteroid dehydrogenase-isomerase activity in rat adrenocortical cells by corticotropin. AB - The regulation of delta 5-3 beta-hydroxysteroid dehydrogenase-isomerase (3 beta HSD) was studied in primary cultures of rat adrenocortical cells. In the absence of ACTH, this enzymic activity was found to decay with a half-life of 3.1 days, which was similar to the half-life of the enzyme activity induced by ACTH in vitro (3.5 days). The increase in 3 beta-HSD activity was highly specific for ACTH and dibutyryl cAMP; the activity was not increased by other hormones known to affect adrenocortical growth or function. The induction of 3 beta-HSD activity by ACTH or dibutyryl cAMP required a lag period of approximately 4 h and was dependent on RNA and protein syntheses. The increase in 3 beta-HSD activity observed after ACTH treatment was not a result of ACTH-induced inhibition of degradation of the enzyme, nor was it due to the synthesis of a soluble intermediate which could directly activate the enzyme. ACTH stimulated the incorporation of [35S]methionine into a protein associated with 3 beta-HSD activity detected on polyacrylamide gels after electrophoresis of Triton X-100 extracts of adrenocortical cells. The induction of this protein by ACTH was inhibited by actinomycin D. A protein band of a partially purified preparation of rat adrenal 3 beta-HSD was found to comigrate with the ACTH-induced protein on sodium dodecyl sulfate-polyacrylamide gel. These results suggest that ACTH caused the de novo synthesis of 3 beta-HSD by a mechanism dependent on RNA synthesis. PMID- 6284481 TI - The mechanism of hypercortisolemia in the squirrel monkey. PMID- 6284482 TI - Metyrapone and antioxidants are required to maintain aldosterone synthesis by cultured bovine adrenocortical zona glomerulosa cells. PMID- 6284483 TI - Nuclear 3,5,3'-triiodothyronine receptors in rabbit lung: characterization and developmental changes. PMID- 6284484 TI - Bioactive and immunoreactive adrenocorticotropin in normal equine pituitary and in pituitary tumors of horses with Cushing's disease. AB - Equine Cushing's disease is caused by hypersecretion of ACTH by hyperplasia or adenomas of pars intermedia (PI) cells, in contrast to human Cushing's disease, which is caused by hyperplasia or adenomas of pars distalis (PD) ACTH-secreting cells. We assayed both bioactive and immunoreactive (IR) ACTH in two normal equine pituitary glands and in the PD, PI, and pars nervosa of four such glands, as well as in the PI adenomas of five horses with Cushing's disease. In normal horse pituitaries, as in those of other species, most of the bioactive and IR ACTH was found in PD, much less in PI, and only traces in pars nervosa. In PI adenomas of horses with Cushing's disease, bioactive ACTH concentrations were similar to those in normal PI, but the total tumor content of bioactive ACTH exceeded that of normal whole pituitary. IR-ACTH concentrations were even higher in PI tumors, suggesting that some of the tumor ACTH was biologically inactive. Plasma IR-ACTH, which, like the PI adenoma tissue, presumably included a major fraction of bioactive ACTH, was greatly elevated in five horses with Cushing's disease and would account for the adrenal hyperplasia and hyperfunction observed in these animals. PMID- 6284487 TI - Fate of human chorionic gonadotropin bound to rat corpora lutea. AB - The current studies were designed to ascertain the fate of hCG bound to rat corpora luteal cell receptors. Graded doses of highly purified hCG (CR119), ranging from 0.1-10.1 micrograms, were injected. Groups of pseudopregnant rats received iodinated hCG, unlabeled hCG, or both. Supraphysiological levels of hCG were used to enhance the internalization of hCG and its receptors. When ovarian membrane pellets (48,000 X g) were subjected to continuous sucrose density ultracentrifugation, two different ovarian membrane fractions (F1 and F2) bound hCG. Although an increase in hCG binding to the F2 membrane fraction was observed between 1-6 h after a single 0.1-microgram [125I]iodo-hCG injection, no subsequent enhanced binding to that fraction was observed. However, the F1 fraction bound at least 3 fold more hCG than did the F2 fraction 1, 6, 12, and 24 h after the injection of 0.1 micrograms [125I]iodo-hCG. When groups of animals were injected with 0.1, 1.0, or 10.0 micrograms unlabeled highly purified hCG, peak serum, ovarian plasma membrane, and ovarian intracellular hCG concentrations were observed at different times after hormone injection and suggested the progressive transfer of hCG from serum to ovarian fractions in a time- and dose dependent relationship. Although no intracellular hCG was detected until 60 min after the single 0.1-microgram injection of hCG, both serum and membrane-bound levels were measurable within 15 min of that injection. From these observations, we suggest that ovarian intracellular hCG does not reflect significant contamination with serum or interstitial fluid or from significant dissociation of membrane-bound hCG during tissue handling. Finally, when intracellular hCG was subjected to continuous sucrose density gradient ultracentrifugation, a single major 135I peak was observed, and this comigrated with [125I]iodo-hCG. Our interpretation of the foregoing observations is that the major intracellular form of hCG is not receptor bound. PMID- 6284486 TI - Rat granulosa cell differentiation: an in vitro model. PMID- 6284485 TI - The interaction of cultured thyroid cells of early bovine embryos with thyrotropin and thyroglobulin. AB - We have prepared primary thyroid cell cultures of early bovine embryos from the first trimester of pregnancy in order to study the ontogeny of their interaction with TSH and thyroglobulin (Tg). The ability of these cells to synthesize and secrete Tg, as well as the trophic effect of TSH on the organization of the thyroid cells, were also investigated. To determine the maturation of these functions we prepared fluorescent conjugates of TSH, Tg, and anti-Tg antibodies, and visualized their interaction with the thyroid epithelial cells. Our study shows that the ability to bind TSH and Tg exists as early as the gestational age of 3 cm crown-rump length (CRL; 40 days) but does not develop linearly with embryonic age. Thus, there is a significant increase in the percentage of Tg binding cells at 12 cm CRL, when colloid is first noticed in vivo, and a considerable elevation in TSH-binding cells around 15 cm CRL, when thyrotropic cells and TSH secretion from the fetal pituitary are first evident. Tg-containing cells and the ability to secrete Tg are observed at about 20 intrauterine days. The three thyroidal properties probably develop independently since only part of the Tg-containing cells bind Tg or TSH, and a significant proportion of the cells that exhibit Tg-binding do not bind TSH. The results support the notion that TSH is essential for the formation of follicle-like structures and effects the organization of thyroid cells into a functional structure in vitro from the late precolloidal stage. PMID- 6284488 TI - On the primary site of action of estrogens and androgens in the regulation of hepatic prolactin receptors. AB - The aim of the present study was to identify the primary sites of action of estrogens and androgens in the regulation of hepatic PRL receptors in the rat. Implantation of estradiol benzoate in the pituitary region of male rats caused a feminization (i.e. an increase) of PRL receptors to a concentration typical of that found in female rats. Estrogen implanted in the paraventricular region of male rats was less effective in causing such a feminization. Subcutaneous implantation of estrogen did not affect the PRL receptor concentration. It is concluded that estrogen induces PRL receptors via an action at the hypothalamo pituitary level, possibly directly on the pituitary. The PRL receptor-suppressive action of sc injected testosterone was also exerted by the synthetic androgen R1881 (17 beta-hydroxy-17 alpha-methylestra-4,9,11-trien-3-one). However, anterior hypothalamic deafferentation rendered both male and female rats insensitive to this action of R1881. It is concluded that an intact hypothalamo pituitary unit is required for the PRL receptor-suppressive action of R1991. It is possible that the site of action of androgens is in the rostral hypothalamus or in adjacent areas of the brain. PMID- 6284489 TI - Expression of glucagon receptors on T- and B- lymphoblasts: comparison with insulin receptors. AB - Activation of T- and B-lymphocytes by a variety of immunological stimuli has been reported to induce specific insulin receptors. The purpose of the present work was to determine whether glucagon receptors are also induced in activated cells. Studies of glucagon and insulin receptors were carried out using normal human mononuclear cells activated by phytohemagglutinin or T-cell growth factor (TCGF), as well as established B- and T-lymphoblastoid cell lines. With phytohemagglutinin, glucagon and insulin binding increased 15- and 36-fold, respectively, and peaked after 5 days in parallel with the rise in thymidine incorporation. Increased binding was associated with an increase in the number of receptors, most marked for insulin, though affinity for the insulin receptor was decreased. Normal human mononuclear cells cultured with TCGF showed an early modest rise in insulin binding due to increased receptor number, without a change in affinity, and a striking and progressive rise up to 50-fold in glucagon binding due to both increased receptor number and affinity. The differences in receptor response to these T-cell mitogens suggest that TCGF selects out a T lymphoblast subset with very high glucagon receptors. B- and T-lymphoblastoid cells showed patterns of glucagon and insulin receptors that appear to be characteristic for each cell type. Glucagon binding was 7-fold higher (P less than 0.01), while inulin binding was 7-fold lower (P less than 0.01) in T- vs. B lymphoblastoid cells. T-Cell lines had twice the number of glucagon receptors, whereas B-lines had 4-fold the number of insulin receptors, with much greater affinity for insulin compared with T-line insulin receptors. Induction of both insulin and glucagon receptors on activated lymphoblasts suggests that these receptors may play a significant role in cell function. PMID- 6284490 TI - Dissociation between the magnitude of nuclear binding and the biopotency of glucocorticoids. AB - After 5 days of incubation with 10 nM tritiated dexamethasone, the amount of glucocorticoid displaceably-bound by the ACTH-secreting, glucocorticoid responsive AtT-20 mouse pituitary tumor cell decreases by 70-80%. This decrease takes place in both the cytosolic and nuclear fractions. Although one might predict that there should be a decrease in agonist efficacy associated with this loss of nuclear binding, there was not; long-term incubations with dexamethasone revealed that the AtT-20 cells showed no tendency to resume secreting ACTH. Hence, although there was a dramatic decrease in the amount of nuclear-bound steroid, there was not a resulting decrease in the steroid's biopotency. These results suggest that the agonist-induced decrease in receptor content is either a means of removing redundant nuclear receptors, or a new, previously unrecognized, step in the mechanism of glucocorticoid hormone action. PMID- 6284491 TI - UP-regulation of lactogenic receptors -- an immunological artifact? AB - We examined whether injection of heterologous hormones for more than one week might evoke a humoral immune response which would simulate receptor induction. Male rats were injected daily for ten days with human growth hormone (hGH) or ovine prolactin (oPRL), and binding of 125I-hGH and 125I-oPRL was examined in serum and in membranes from liver and lung. Specific binding of 125I-hGH and 125I oPRL increased in the sera of hGH- and oPRL-injected animals, respectively. A marked increase in hGH but not oPRL binding also occurred in crude membrane preparations of tissues from hGH-injected rats. Similarly oPRL but not hGH binding increased in tissues of PRL-injected animals. Furthermore, binding activity solubilized from liver membranes of hormone-injected rats was precipitated with Staphylococcus aureus (protein A) indicating that the induced binding sites were immunoglobulin-like. Hence apparent up-regulation of lactogenic receptors following long-term treatment with heterologous hormones may be due to generation of anti-hormone antibodies. PMID- 6284492 TI - Carcinoembronic antigen and endocrine response in a patient with Sipple's syndrome following surgery. AB - Circulating carcinoembryonic antigen (CEA), calcitonin (CT) and adrenocorticotropic hormone (ACTH) and levels in tissue extracts of a patients with Sipple's syndrome were measured simultaneously. Biofocal medullary carcinoma of the thyroid gland (MCT), left adrenal phaeochromocytoma and ectopic para aortal phaeochromocytoma had been removed in the first operation. Total thyroidectomy was performed thereafter. Evidence has been presented showing that (1) the first operation lowered the elevated serum levels of CEA and CT, and levels of CEA and CT in tissue extracts of MCT were considerably higher than those of tumor-uninvolved thyroid tissue and phaeochromocytomas; (2) markedly elevated levels of plasma ACTH were normalized by the first surgery and, in addition, immunoreactive ACTH and beta-melanocyte stimulating hormone (beta-MSH) were detected in the phaeochromocytomas removed. This is a documented case of the simultaneous production of CT and CEA by MCT associated with ectopic secretion of ACTH and beta-MSH from the phaeochromocytomas without manifesting clinical features of Cushing's syndrome and is based on the findings referred to above. PMID- 6284493 TI - A rapid method for the separation of rat pancreatic islets from collagenase digested pancreas using Percoll. AB - The isolation of pancreatic islets from collagenase-digested Wistar rat pancreas was shown by the sedimentation method at a unit gravity using Percoll solution with a density of 1.041 g/ml. The density of digested exocrine tissues was in the range of 1.013-1.041 g/ml, while that of purely isolated islets was in the narrow range of 1.066-1.075 g/ml. More than a hundred islets were obtained freely from each rat pancreas without any gross contamination of digested exocrine tissues. A significant increase was observed in glucose-stimulated insulin release from islet isolated with Percoll in the same pattern as that without Percoll. In addition to the well preserved morphology and function of pancreatic islets isolated by Percoll, the simplicity of the technique strongly commends the usefullness of this method. PMID- 6284494 TI - A case with glucagonoma syndrome--heterogeneity of glucagon and insulin. AB - The heterogeneity of glucagon and insulin in plasma and tissue extracts from a 57 year-old female with glucagonoma syndrome with surgically and autopsy verified islet-cell tumors was studied by Bio-Gel P-10 filtration. The preoperative plasma immunoreactive glucagon (IRG) level was 20.2 ng/ml, and plasma glucagon-like immunoreactivity(GLI) 25.8 ng/ml. The column chromatography of the preoperative plasma revealed three or four IRG components and four GLI components. Among these, peak II, the large glucagon immunoreactivity (LGI) peak, considered a candidate for proglucagon, was prominent, along with peak III. The resected metastatic liver tumor contained an enormous amount of IRG and an appreciable amount of immunoreactive insulin (IRI), indicating that the elevated plasma IRG was mainly of tumor origin. The IRG pattern of the tumor tissue extract revealed a small quantity of IRG in peaks I and II, and a large amount in peak III; control pancreatic tissue extract manifested a similar elution pattern. The IRI elution pattern of the tumor tissue extract revealed two major IRI peaks which migrated close to the elution volume of cytochrome C and insulin, respectively. This is a quite different pattern from the control pancreatic tissue extract in which the RI peak was localized in the elution volume of the insulin. We conclude that the present metastatic liver tumor produced not only enormous amounts of glucagon but heterogeneous peptides which contained immunological insulin determinants within their. PMID- 6284495 TI - Inhibitory effects of 1 alpha, 25-dihydroxycholecalciferol on parathyroid hormone secretion in rats. AB - In an attempt to study the influence of vitamin D metabolites on PTH secretion, serum calcium and urinary excretion of cAMP were sequentially measured in conscious perfused rats, and the effects of a single iv injection of the metabolites on these parameters were examined. Four hours after the administration of 0.25 microgram/kg (0.6 nmol/kg, probably a physiological dose) of 1 alpha, 25-dihydroxycholecalciferol [1 alpha, 25 (OH)2D3], the urinary excretion of cAMP decreased to a level compatible with that of parathyroidectomized rats (approximately 60% of the initial value; P less than 0.05) and this level was sustained for nearly 24 h. Serum concentrations of calcium (total and ionized) did not change. In parathyroidectomized rats which were continuously infused with bovine PTH (1 U/h), the vitamin D metabolite had no significant effect on the urinary excretion of cAMP. 24 R, 25 dihydroxcholecalciferol (12.5 microgram/kg) had no significant effect either on the urinary excretion of cAMP or on serum calcium. These results suggest that in rats, a physiological dose of 1 alpha, 25(OH)2D3 inhibits PTH secretion without causing a significant rise iu serum calcium, reflecting a feed-back mechanism between active vitamin D metabolite, 1 alpha, 25(OH)2D3 and the parathyroid glands. PMID- 6284496 TI - Changes in distribution of molecular weight forms of biologically active and immunoreactive adrenocorticotropic hormone after adrenalectomy in rat anterior pituitary. AB - The concentration of ACTH in extracts of rat anterior pituitary was measured by both radioimmunoassay and bioassay at different stages following adrenalectomy. Both types of ACTH activity decreased the day immediately following adrenalectomy but increased gradually afterwards. Immunological ACTH activity increased to 250% of the control value and biological ACTH activity increased to 490% of control value 3 weeks after adrenalectomy. The increase in biological ACTH activity occurred earlier, and the rate of increase was greater, than that of the immunological ACTH activity. The distributions of molecular weight forms of ACTH in extracts of anterior pituitary lobes was determined by gel filtration. Three molecular weight forms of immunoassayable ACTH were detected. Biological ACTH activity appeared in the 2nd and the 3rd peaks. A striking change was observed after adrenalectomy in the distribution of biologically active forms of ACTH. The ratio of biological ACTH activity to immunological ACTH activity in each peak changed at various stages after adrenalectomy. This indicated the heterogenous nature of the ACTH included in each peak. At 2 and again at 3 weeks, biological activity markedly increased until it exceeded the immunological ACTH activity in the 2nd peak. Dexamethasone had little influence on the elution profile of either immunoassayable and biologically active ACTH in gel filtration. Adrenalectomy may possibly have an effect on the intracellular posttranslational processing of ACTH precursors which leads to the development of biological ACTH activity. PMID- 6284497 TI - The radioactive labeling of polypeptide hormones with myeloperoxidase in solid state comparison with chloramine T method. PMID- 6284498 TI - Abnormalities in monoamine levels in the central nervous system of the genetically epilepsy-prone rat. AB - Norepinephrine, dopamine, and 5-hydroxytryptamine concentrations were determined in the central nervous systems of genetically epilepsy-prone rats (GEPR) and in control rats. Norepinephrine concentrations were abnormal in all major areas of the central nervous system of the GEPR, with decrements existing in the telencephalon, hypothalamus-thalamus, midbrain, pons-medulla and spinal cord. An increment in the concentration of this neurotransmitter existed in the cerebellum. Dopamine concentrations were normal in all areas of the GEPR brain. Abnormalities in 5-hydroxytryptamine concentrations were also present in the GEPR. They were exclusively decrements and occurred in the telencephalon, hypothalamus-thalamus, midbrain, and pons medulla. Concentrations of this neurotransmitter were normal in the cerebellum and spinal cord. Coupled with our earlier pharmacologic data, these observations support our concept that noradrenergic and/or 5-hydroxytryptaminergic decrements are etiologically important in seizure susceptibility in the GEPR. The lack of abnormalities in brain dopamine concentrations strengthens our hypothesis that dopaminergic transmission does not regulate seizure susceptibility in this model. PMID- 6284499 TI - Sister chromatid exchange response of human diploid fibroblasts and Chinese hamster ovary cells to dimethylnitrosamine and benzo(a)pyrene. AB - In the search for relevant assays for mutagenicity testing, considerable attention has been given to the use of mammalian cells in vitro and the incorporation of metabolic activation in the protocol. Chinese hamster ovary (CHO) cells are commonly chosen as the target cells for cytogenetic tests because of their excellent growth characteristics and long lifespan in culture. However, there may be cellular factors affecting the uptake, metabolism, and repair of damage which are not the same in all cell lines. The response of CHO cells and three human diploid fibroblast strains (IMR-90, WI-38, S-3299) to benzo(a) pyrene (BP) and dimethylnitrosamine (DMN) were compared using sister chromatid exchange (SCE) analysis as a measure of genetic damage. For both BP and DMN the human cells and the CHO cells showed dose-response slopes that were significantly different from zero, except CHO cells treated with BP for 1 hr and S-3299 cells treated with DMN. Whereas human and CHO cells showed similar dose-responses to BP and the three human cell strains had similar dose-responses to BP and DMN, the dose-response of the human cells to DMN was statistically less significant than that of CHO cells. Reducing the duration of chemical treatment in CHO cells had no effect on the slope of the dose-response curves for BP or DMN. The observed differences between human and CHO cells may reflect differences in the fate of metabolic intermediates of DMN. PMID- 6284500 TI - Sister chromatid exchange induction in two cell lines. AB - Sister chromatid exchange (SCE) induction by the procarcinogen benzo (a)-pyrene (BP) was studied using two established cell lines. Chinese hamster ovary (CHO) and Chinese hamster lung (Don) cells were exposed to varying concentrations of BP in the presence and absence of a rat liver metabolizing system. Results showed a significant difference in the abilities of the two cell lines to induce SCE in the absence of liver homogenate. These experiments demonstrate that Don cells are capable of generating active metabolites from biologically inactive procarcinogens presumably due to an inherent metabolizing system which is deficient or absent in CHO cells. PMID- 6284501 TI - Endonucleases in yeast mitochondria: apurinic and manganese-stimulated deoxyribonuclease activities in the inner mitochondrial membrane of Saccharomyces cerevisiae. AB - An apurinic endonuclease activity has been characterized in yeast mitochondrial. It is dependent on Mg2+, stimulated by about 50% in the presence of 50 mM NaCl and inhibited at higher NaCl concentrations. It is located in the inner mitochondrial membrane and requires high concentrations of detergent (1.5-3% Triton X-100) to be extracted. The same treatment extracts several other endonuclease activities: the two Mg2+-dependent endonuclease activities cleaving double-stranded DNA at pH 7.5 and 5.4 respectively, the ethidium-bromide stimulated endonuclease activity, the endonuclease activity cleaving single stranded DNA at pH 7.l5 [Jacquemin-Sablon et al. (1979) Biochemistry, 18, 119 127], and a manganese-stimulated deoxyribonuclease activity cleaving double stranded DNA at pH 7.5 which has been discovered during the present work. Another endonuclease activity cleaving double-stranded DNA at pH 7.5 in the presence of Mg2+, slightly stimulated by low NaCl concentrations and inhibited by ethidium bromide is extracted from the membrane pellet remaining after the treatment with 1.5% Triton X-100 by a second treatment with 1.5% Triton X-100 plus 1 M KCl. The presence in the mitochondrial membrane of this apurinic endonuclease activity indicates that, like nuclear and prokaryotic DNA, yeast mitochondrial DNA is also subject to specialized repair systems. PMID- 6284503 TI - 1H NMR studies of eukaryotic cytochrome c. Resonance assignments and iron hexacyanide-mediated electron exchange. AB - 1H NMR resonance assignments in the spectra of horse, tuna, Neurospora crassa and Candida krusei cytochromes c are described. Assignments have been made using NMR double-resonance techniques in conjunction with electron-exchange experiments, spectral comparison of related proteins, and consideration of the X-ray structure of tuna cytochrome c. Resonances arising from 11 residues of horse cytochrome c have been assigned. PMID- 6284502 TI - Affinity labelling and characterization of the ppp(A2'p)nA-dependent endoribonuclease from different mammalian sources. AB - The ppp(A2'p)nA-dependent endoribonucleases from a number of different mammalian sources have been investigated. The enzyme from reticulocyte lysates shows optimal activity of 50-150 mM KCl and requires the presence of Mg2+. Whilst the enzyme is inactivated after passage of reticulocyte lysates through Sephadex columns in the absence of ATP, it retains full activity provided ATP is included in the column buffer. The activity of the partially purified nuclease was unaffected by the addition of reticulocyte RNase inhibitor, which, in contrast, effectively inhibited other endogenous endonucleases. The ppp(A2'p)nA-dependent Rnase co-purified with a ppp(A2'p)nA-binding protein and with a protein which could be specifically covalently labelled with an oxidised radioactive analogue of ppp(A2'p)nA. This covalent labelling could be carried out either with the partially purified RNase or in crude extracts from rabbit reticulocytes, mouse Krebs and Ehrlich ascites tumour cells and human lymphoblastoid (Daudi) or HeLa cells. In each case the affinity labelled protein migrated to a position corresponding to a apparent molecular weight of about 85 000 on electrophoresis on dodecylsulphate/polyacrylamide gels. In all cases labelling could be prevented by the addition of an excess of unlabelled ppp(A2'p)nA but not, for example, by a similar excess of the biologically inactive dimer ppp(A2'p)'A. It is concluded that the RNase and ppp(A2'p)nA binding activities are likely to reside in the same molecule. PMID- 6284504 TI - 1H NMR studies of the electron exchange between cytochrome c and iron hexacyanides. Definition of the iron hexacyanide binding sites on cytochrome c. AB - Binding of [Fe(CN)6]3-, [Cr(CN)6]3-, [Co(CN)6]3- and [Cr(C2O4)3]3- to horse, tuna and Candida krusei cytochromes c has been studied by high-resolution 1H NMR spectroscopy. All the reagents bind at the same sites. There are at least two binding sites, and probably three, on horse cytochrome c at pH 7. One of the sites is only a weak binding site and is far from the haem group, whereas the other site(s) is(are) at the haem crevice. Ka for binding of [Fe(CN)6]3- to trimethyllysine-72 of C. krusei ferricytochrome c is 140 +/- 15 M-1 at 27 degrees C and pH 7. PMID- 6284505 TI - Structural, functional and conformational properties of rat hemoglobins. AB - The oxygen equilibrium properties of rat total hemoglobin show pH dependence. Thus oxygen affinity and cooperativity, which are significantly reduced at pH 6.0, show increase with increasing pH. Conformational studies using nitrosyl derivatives indicate that in rat nitrosyl hemoglobin the R to T equilibrium is shifted towards the T state in going from pH 7.0 to pH 6.0. Under similar conditions human hemoglobin A shows no significant changes in cooperativity or conformation. These results indicate that a destabilized oxy structure (R) exist in rat hemoglobins at pH 6.0. Alterations in the heme pockets and alpha 1 beta 2 contact points could lead to these structural characteristics. Crystallization of rat hemoglobins is dependent on an oxy-like structure since all liganded ferrous derivatives and methemoglobin form crystals in the pH range of 7.0-8.0. Deoxygenation of oxyhemoglobin crystals or addition of inositol hexakisphosphate to nitrosyl or methemoglobin derivatives result in solubilization of the crystals. Thus the quaternary and/or tertiary structural changes involved in the transition of a R to T state disrupt the crystal structure. The precise positioning of the complementary sites involved in the interaction of amino acids between rat hemoglobin tetramers seems to occur only when the molecules are in the quaternary or tertiary R conformation. This is in contrast to the polymerization of human deoxyhemoglobin S, where gel formation occurs only in the T state. PMID- 6284507 TI - Plasminogen activator released as inactive proenzyme from murine cells transformed by sarcoma virus. AB - We have previously reported the purification of a plasminogen-activating serine protease with an approximate Mr of 48 000 from sarcoma-virus-transformed murine cells. We now report that under serum-free conditions the enzyme is released from the cells in an inactive form. After affinity chromatography with 4 aminobenzamidine-cellulose, ion-exchange chromatography and gel filtration, the proenzyme could be obtained from culture fluid as a pure, homogeneous protein as evaluated by polyacrylamide gel electrophoresis with sodium dodecylsulphate. Proenzyme was quantitatively converted to active enzyme by incubation with catalytic amounts of plasmin. Analysis by polyacrylamide gel electrophoresis with sodium dodecylsulphate under reducing and non-reducing conditions showed that the inactive form consisted of a single polypeptide chain with an Mr of approximately 48 000, while the active form consisted of two chains with Mr values of approximately 18 000 and 29 000 , held together by one or more disulphide bridges. The active-site reagent diisopropylfluorophosphate in radiolabelled form was incorporated into the 29 000-Mr chain of the active enzyme, but not into the inactive form. These findings provide conclusive evidence for the existence of an inactive proenzyme to this plasminogen activator and thus demonstrate and additional step in a cascade-like reaction leading to extracellular proteolysis. Regulatory as well as methodological implications of this findings are discussed. PMID- 6284506 TI - The pressure dependence of the spin equilibrium in camphor-bound ferric cytochrome P-450. AB - The spin equilibrium of camphor-bound ferric cytochrome P-450 has been measured between 1-1000 bar (10(5)-10(8) Pa). Increasing pressure shifts the absorption spectrum from the high-spin form at 392 nm to the low-spin form at 417 nm. The molar volume change for the spin states delta V = -RT delta ln Ke/ delta P and the equilibrium coefficient Ke = [high spin]/[low spin] depend on the solvent conditions. At pH 5.6 the equilibrium coefficient at 1 bar, K1 = 0.5 and delta V = 312 cm3/mol. A sample with 10 mM KCl at pH 7 has K1 = 7.0 amd delta V = 52 cm3/mol. Solvent changes producing a larger K1 also result in a larger delta V which ranged over 16-74 cm3/mol. The correlation can be approximated as delta V = 36 + 18 log K1, which implies that there is a pressure, 3000 bar, for camphor bound ferric cytochrome P-450 at 4 degrees C, at which the changes in delta V are compensated by the other thermodynamic parameters leaving Ke independent of the solvent conditions. Although the protein is not stable about 1000 bar for most sample conditions, the extrapolated log Ke versus pressure curves for all sample conditions intersect near 3000 bar. Camphor-bound cytochrome P-450 appears to be a rather flexible protein, having a low denaturing pressure, a large volume change, and a high sensitivity to the protein environment. PMID- 6284509 TI - Perspectives of adenylate kinase activity and glutathione concentration in cerebrospinal fluid of patients with ischemic and neoplastic brain lesions. AB - Adenylate kinase (AK) activity and glutathione concentration were measured in cerebrospinal fluid (CSF) on three different occasions after the ictus in 16 patients with cerebral infarction. All patients displayed an unambiguous AK activity in their CSF within 48 h after the ictus while glutathione was present in 10 of the 16 patients. The mean AK activity among these patients was reduced by about 60% after 1 week and an additional smaller reduction took place after 3 4 weeks. The glutathione concentration in CSF of these patients showed a similar curve profile with time. The 9 patients who showed clinical signs of improvement had significantly lower values on all occasions for both AK and glutathione than those who did not improve or deteriorated. A corresponding reduction in AK activity with time was not observed in 3 patients with cerebral malignancies. The possibility was excluded that the enhanced AK activity in CSF was the result of an increased transfer of the enzyme from the plasma compartment. Instead of causal relationship was suggested between the appearance of AK in CSF and a disturbed brain cell metabolism. Repeated analyses revealed different curve profiles for ischemic and neoplastic brain lesions. The importance of sampling and handling the CSF material under standardized conditions is emphasized. PMID- 6284508 TI - Ultrastructural pathology of human lymphocytes in lysosomal disorders: a contribution to their morphological diagnosis. AB - Ultrastructural examination of peripheral lymphocytes was performed in 28 cases of various lysosomal diseases, including infantile, late infantile and juvenile neuronal ceroid-lipofuscinoses (NCL), mucopolysaccharidoses (MPS), juvenile and adult metachromatic leukocystrophies (MLD), GM1-gangliosidosis, one patient with presumed mucolipidosis type IV, mucolipidosis type III, and glycogenosis type II. Based on our own observations on the ultrastructure of lymphocytes in lysosomal disorders, our results may be divided into the following 3 groups: 1. pathological findings with specific inclusions: each type of NCL, presumed mucolipidosis type IV, glycogenosis type II; 2 pathological findings with vacuoles: types I-H, II, III-A and III-B, IV, VI-A and VI-B of MPS, GM1 gangliosidosis; 3. apparently no pathological findings: juvenile and adult MLD, mucolipidosis type III, GM2-gangliosidosis, Gaucher disease. These results led us to conclude that morphological investigations utilizing lymphocytes do not always offer sufficient diagnostic information although easy accessibility favors diagnostic ultrastructural studies of lymphocytes. Such morphological studies should be supplemented by diagnostic biochemical methods. PMID- 6284510 TI - Postganglionic cholinergic dysautonomia: report of muscle findings in 1 case. AB - Heretofore unreported findings in skeletal muscle in a 4th case of pure postganglionic dysautonomia are described. They consist of variation in fiber size, Z line and myofilamentous disorganization, intracytoplasmic inclusions and honeycomb formations. The most striking alterations, however, was excess lipid accumulation without concomitant mitochondrial alterations. Sural nerve histology and cytology were normal. The mechanism of lipid accumulation in skeletal muscle of patients with dysautonomia remains unexplained. PMID- 6284511 TI - Progression of endocrinological and neurological dysfunction in adrenoleukodystrophy. Endocrinological evidence of an affection of the adrenal cortex independent of the cerebral disorder. AB - In a boy suffering from adrenoleukodystrophy (ALD) the function of the adrenal cortex and pituitary gland was followed prospectively. At the onset of the disease an autonomic hyperfunction of the adrenal cortex was shown. Subsequently the secretion of cortisol dropped to low levels despite high plasma levels of ACTH. The results suggest a primary disorder of the adrenal cortex in ALD, which up till now could only be postulated on anatomical evidence. With the use of computerized tomography and electroencephalography abnormalities characteristic for ALD were shown. The progression of the disease was followed using these noninvasive methods of examination. PMID- 6284512 TI - Xeroderma pigmentosum: neurological, neurophysiological and morphological studies. AB - In 3 cases of xeroderma pigmentosum showing signs of peripheral neuropathy, electrophysiological studies were made. In one of them, a sural nerve biopsy was performed. MCV obtained from the lower limbs were moderately reduced in all cases, and SCV could be obtained in only 1 case from the median nerve. Auditory brain stem responses were not obtainable in all cases. Sural nerve biopsy revealed a marked decrease of myelinated fibers and also suggested severe damage in both myelinated and unmyelinated fibers. Teased fiber study showed myelin ovoids and indicated axonal degeneration. Sensory nerves including the acoustic nerve may suffer damage earlier than the motor nerve. In all cases CT scans revealed brain atrophy and thickening of the skull. PMID- 6284513 TI - DMSA scan and the prediction of recovery in obstructive uropathy. PMID- 6284514 TI - Early growth of experimental tumors and the onset of concomitant immunity. PMID- 6284515 TI - Retinoic acid binding proteins and steroid receptor levels in human breast cancer. AB - Samples of 31 human breast cancer tissues were examined for the presence or absence of cytosolic retinoic acid binding proteins (cRABP), estrogen receptors (ER) and progesterone receptors (PR). cRABP were detected as specifically sedimenting 2S components on sucrose density gradient. Specific binding under the 2S region for cRABP and under the 8S region for ERR and PR were determined. The levels of cRABP ranged from less than 1 pmol to 14 pmol per mg of cytosol protein. Tumors containing less than 3 pmol cRABP were designed as low cRABP; tissues were considered ER- and PR- when the receptor levels were less than 10 fmol/mg protein. Approximately 60% of primary and metastatic tumors examined contained high cRABP. Tumors with well differentiated histopathology exhibited increased levels of cRABP. There was no significant correlation between the presence or absence of steroid receptors and cRABP in human mammary tumors. PMID- 6284517 TI - Rous sarcoma virus-induced tumors in mice--I. Macrophage-mediated natural cytotoxicity. AB - Lymphoid cells from normal untreated A.SW mice showed a cell-mediated cytotoxicity (CMC) towards Rous sarcoma virus-induced tumor lines in a 48-hr microcytotoxicity assay in vitro, using 3[H]-thymidine prelabeled tumor cells. The tissue distribution of this natural CMC shows high levels of activity in the peritoneal cavity, intermediate levels in bone marrow and spleen, and low levels in lymph node, peripheral blood and thymus. Natural CMC of unstimulated resident peritoneal cells against tumor cells was also found in several strains of mice. This was not dependent on the donor's age, since similar levels appeared in mice aged 1-40 weeks. Passage through nylon wool columns or adherence to the plastic surface of resident peritoneal cells removed cytotoxic activity. By contrast, adherent cell displayed significant CMC levels. Treatment with carbonyl iron powder and magnet decreased peritoneal cell cytotoxicity when performed once and abolished it when repeated. All these results suggest that in mice, natural cytotoxicity against Rous sarcoma virus-induced tumors is primarily mediated by macrophages or macrophage-like cells. PMID- 6284516 TI - Concomitant secretion of calcitonin, beta-endorphin and ACTH from medullary thyroid carcinoma in vivo and in vitro. AB - The present study was performed to investigate the possible synthesis of a common precursor molecule for calcitonin (CT), adrenocporticotropin (ACTH) and beta lipotropin (beta-LPH)/ beta-endorphin (beta-EP) by the human medullary thyroid carcinoma (MTC). In a patient with MTC but without Cushing's syndrome, the response of plasma CT, ACTH and cortisol levels to a calcium infusion, lysine vasopressin (LVP) and dexamethasone were measured. A parallel increase of these hormones in response to calcium and LVP was seen, while there was a paradoxical increase of CT during dexamethasone infusion. Incubation of MTC fragments obtained at surgery showed a significant correlation of the secretion of CT, ACTH and beta-LPH/ beta-EP in response to calcium, LVP and dexamethasone. The concomitant release of these hormones in vivo and in vitro could be compatible with the synthesis of a common precursor molecule for CT, ACTH and beta-LPH/ beta EP in MTC, although this was not substantiated by gel-chromatography of the tumor extract. Corticotropin releasing factor, a regulator of the normal processing of pro-opiococorticoid precursor molecule in the anterior pituitary gland, is also able to activate ACTH, beta-LPH/ beta-EP and calcitonin secretion from the malignant C-cell of the thyroid. PMID- 6284518 TI - Gastroscopic and pharmacokinetic evaluation of a new pivmecillinam tablet. AB - Three different pivmecillinam preparations, a conventional 200 mg tablet (P tablet) and two new formulations containing respectively pivmecillinam 200 mg and 400 mg plus Avicel (microcrystalline cellulose) as a disintegrator (PA tablet), were compared in vitro and in a gastroscopic study in 8 healthy volunteers. Disintegration of the PA tablet was significantly more rapid both in vitro and in the stomach. Following disintegration, the content of the PA tablet was spread over a larger area of the gastric mucosa (1088 mm2) than was observed with the P tablets (408 mm2). Three of the 8 volunteers taking the P tablet developed hyperaemia, interstitial bleeding or erosions of the mucosa of the stomach. No such reactions were seen with the PA tablets. Serum concentrations of mecillinam following ingestion of pivmecillinam tablets were determined in three groups of subjects; fasting volunteers, both supine and ambulant, and in ambulant subjects who took the preparation with a light meal. There was a tendency for the new PA tablets to produce a higher peak serum level as well as greater bioavailability of mecillinam. Administration of the PA tablets with a meal significantly increased the peak serum level and total bioavailability of the drug. On the basis of our observations we recommend adoption of the new PA tablet, because of its quick passage through the oesophagus and its more rapid and complete disintegration in the stomach. PMID- 6284519 TI - Pharmacokinetics of ceftriaxone following intravenous administration of a 3 g dose. AB - The pharmacokinetic parameters of total (bound and unbound) and free (unbound) ceftriaxone in six healthy volunteers after intravenous injection of 39 were compared with low-dose data from a previous study. The dose-dependent behaviour of total drug was considerably more pronounced after the 3 gram dose. In contrast, total body clearance (C1FS = 258 ml/Min), renal clearance (C1FR = 170 ml/min) and volume of distribution (VFD (beta) = 168 1) of free (unbound) drug did not differ from the data reported earlier. There was no significant change in biological half-life (t1/2 (beta) = 7.8 h) or in the fraction excreted unchanged in urine (fu = 0.67). PMID- 6284520 TI - Induction and augmentation of mitogen-induced immune interferon production in human peripheral blood lymphocytes by N alpha-desacetylthymosin alpha 1. AB - Treatment of human peripheral blood lymphocytes with cloned N alpha desacetylthymosin alpha 1 induced interferon production. The kinetics is similar to that of mitogen-induced interferon induction. N alpha-desacetylthymosin alpha 1, in combination with mitogen, augments the amount of interferon produced. This interferon is immune interferon (IFN-gamma) as determined by sensitivity to pH 2, lack of neutralization by antibodies to IFN-alpha or IFN-beta and absence of activity of MDBK cells. Although the mechanism of induction of IFN-gamma by N alpha-desacetylthymosin alpha 1 is unclear, this compound is not mitogenic at concentrations causing IFN-gamma production. These results indicate that thymic factors may also participate in the regulation of IFN-gamma-production. PMID- 6284521 TI - Analysis of DR-like molecules on a marmoset Epstein-Barr virus-induced cell line using a monomorphic anti-human HLA-DR monoclonal antibody. AB - Mouse anti-HLA D region-related (DR) monoclonal antibodies have been found to cross-react with peripheral blood leukocytes from one primate species, the common marmoset (Callithrix jacchus). Immunoprecipitates of radioactively labeled cells extracted from a marmoset Epstein-Barr virus-induced cell line were analyzed by one- and two-dimensional gel electrophoresis and compared with the DR antigens of a human lymphoblastoid B cell line. Two chains of estimated molecular weights of 34 000 (alpha) and 28 000 (beta), similar to the human alpha and beta chains, have been observed in marmoset immunoprecipitates. Additionally, a set of spots located in the same area as the set of invariant spots found in human HLA-DR antigens is shown by two-dimensional gel electrophoresis. Thus, cross-reacting anti-human HLA-DR monoclonal antibodies could be used to analyze the expression and the structure of marmoset DR-like antigens. PMID- 6284522 TI - The choice of opiate receptor subtype by neo-endorphins. AB - The choice of opiate receptor subtype by alpha- and beta-neo-endorphin was studied in isolated preparations. Neo-endorphins had significant inhibitory actions on the electrically evoked contractions of guinea-pig ileum, mouse vas deferens and rabbit ileum as well as on the rabbit vas deferens which had been shown to contain kappa-receptors exclusively. Mr 2266, a relatively specific kappa-receptor antagonist, was more effective than naloxone, a relatively mu receptor antagonist, to antagonize the agonist actions of neo-endorphins in either the guinea-pig and rabbit ileum or in the rabbit vas deferens. By contrast, in the mouse vas deferens, the effectiveness of Mr 2266 to antagonize the agonist actions of neo-endorphins was low and similar to that of naloxone. The potencies of neo-endorphins relative to that of ethylketocyclazocine, a representative kappa-receptor agonist, in the guinea-pig ileum were similar to those in the rabbit ileum but were significant different from those in the mouse vas deferens. The data indicate that neo-endorphins act as kappa-receptor agonists in either the guinea-pig and rabbit ileum or in the rabbit vas deferens while in the mouse vas deferens they act on opiate receptor subtypes other than kappa- and mu-receptors. PMID- 6284524 TI - Influence of morphine dependence on GABA-stimulated benzodiazepine binding to mouse brain synaptic membranes. PMID- 6284523 TI - Presynaptic alpha-adrenoceptors and opiate receptors: inhibition of [3H]noradrenaline release from rat cerebral cortex slices by different mechanisms. AB - The inhibitory effects of morphine and Cd2+ on electrically evoked [3H]noradrenaline release from superfused brain slices were unaffected when release was enhanced by increasing the pulse duration, while the inhibitory effect of noradrenaline and the enhancing effect of phentolamine were diminished. A similar enhancement of [3H]noradrenaline release by 4-aminopyridine reduced the modulatory effects of all drugs examined. Therefore there seem to be different mechanisms for the effect of presynaptic alpha-adrenoceptors and opiate receptors on the availability of Ca2+ for the stimulus-secretion coupling process in noradrenergic nerve terminals. PMID- 6284525 TI - [Met5]enkephalin-Arg-Phe ACTS on vascular opiate receptors. PMID- 6284526 TI - Reserpine and sympathetic denervation cause an increase of postsynaptic alpha 2 adrenoceptors. AB - The changes in alpha-adrenoceptors in rat vasa deferentia after injections of reserpine and 6-hydroxydopamine (6-OHDA) were examined in binding studies with [3H]WB4101 and [3H]clonidine. On intraperitoneal injection of reserpine (0.5 mg/kg body weight per day, 2 days), [3H]clonidine binding sites increased by about 2.0 pmol/g wet weight; the binding sites were not detectable in vasa from control rats. [3H]WB4101 binding sites showed no apparent change in amount. After a single injection of 6-OHDA (30 mg/kg body weight), [3H]clonidine binding sites amounted to about 1.1 pmol/g wet weight. The data suggest that both reserpine treatment and incomplete chemical denervation with 6-OHDA cause an increase of [3H]clonidine binding sites in the postsynaptic region of rat vasa deferentia. The possibility was tested by completely denervating rat vasa deferentia by chemical treatment and operation. The treatments increased the binding sites to about 0.9 and 1.0 pmol/g wet weight, respectively over the control level. The results imply that the binding sites indeed also increased in the postsynaptic area. PMID- 6284527 TI - Depression by chronic electroconvulsive treatment of clonidine hypotherma and [3H]clonidine binding to rat cortical membranes. AB - The effects of chronic electroconvulsive treatment (ECT) on the binding of [3H]clonidine and on clonidine-induced hypothermia were studied in the rat. After 10 consecutive daily treatments we observed a reduction in the hypothermic action of clonidine and a loss of the high affinity binding sites of clonidine, without changes of the low affinity binding sites. The present results are indicative of the down-regulation of alpha 2-adrenoceptors during chronic antidepressive treatment. PMID- 6284528 TI - Induction of angiotensin-converting enzyme with the ACE inhibitory compound MK 421 in rat lung. AB - Having observed that treatment of rats with captopril led to an increased ACE activity in serum and ACE concentration in lungs, we treated female Wistar Kyoto rats for 7 days with the esterified ACE inhibitor, MK-421 (1.0 mg/kg body weight per day), administered by Alzet osmotic minipump. Serum ACE activity decreased by 67% during MK-421 treatment when measured in non-dialyzed serum samples. Removal of the drug by dialysis unmasked a 280% increase of serum ACE activity. ACE concentration of crude lung homogenate increased 134% in MK-421-treated rats and ACE concentration in purified pulmonary plasma membranes increased by 34%. The increase of serum and lung ACE in MK-421-treated rats was similar to that seen in rats treated with captopril, and was probably due to induction of ACE biosynthesis. The mechanisms of this induction are unknown. PMID- 6284529 TI - Cardiovascular changes in response to selective monoamine oxidase inhibition in the rat. AB - Chronic (21 days) treatment with the selective monoamine oxidase (MAO)-A inhibitor clorgyline, but not with the MAO-B inhibitor deprenyl in pithed rats leads to increased blood pressure responses to sympathetic stimulation and intravenous tyramine, and to elevated unstimulated heart rates. No significant changes are observed in plasma catecholamine responses to sympathetic stimulation, nor in beta-adrenoreceptor numbers in heart ventricles. These findings suggest that the hypotensive effects of MAO inhibitors result from central nervous system rather than peripheral nervous system alterations. PMID- 6284530 TI - Opioids increase laryngeal resistance and motoneuron activity in the recurrent laryngeal nerve. AB - [D-Ala2,Met5]enkephalinamide (DAME), [D-Ala2,Leu5]enkephalinamide (DALE) and morphine sulfate (MS) increase activity in the recurrent laryngeal nerve (RLN) within 1 s subsequent to right atrial administration. The activation of the RLN was correlated with a large increase in the resistance to airflow in the in situ isolated larynx. Single unit recurrent laryngeal motoneuron recordings showed that the increase in laryngeal resistance was caused by the continuous activation and recruitment of expiratory motoneurons. Results obtained with opioids mimicked those of phenyldiguanide (20-40 micrograms/kg RA), an agent known to stimulate pulmonary J-receptors. Opioid-induced increases in laryngeal resistance were blocked by bilateral section of the RLN and pretreatment with naloxone (100 micrograms/kg RA). Naloxone had no effect on PDG responses. During the initial period of laryngeal motoneuron activation, the phrenic nerve (PN) was inhibited, however, the activation of the RLN was not dependent upon PN inhibition. It was concluded that stimulation of pulmonary opiate receptors, which may be associated with vagal afferents, elicit a reflex activation of expiratory recurrent laryngeal motoneurons resulting in an increase in laryngeal resistance. PMID- 6284531 TI - Alpha-adrenoceptor control of peroxidase secretion from rat lacrimal gland cells in vitro. PMID- 6284532 TI - Changes in hepatic microsomal membrane fluidity with age. AB - There are changes in the mixed function oxidase enzymatic activities of rat hepatic microsomal membranes with age. However, the protein components of the mixed function oxidase system do not appear to change with age. The purpose of this study was to detect possible changes in the fluidity of the lipid component of the microsomal membrane with age. Hepatic microsomes were isolated by differential centrifugation from uninduced, male CFN rats aged 3, 12 and 26 mo. The microsomal membrane fluidity was measured using electron paramagnetic resonance after incorporation of a 5-nitroxide stearic acid spin label into the membrane. The order parameter S decreased with age from 0.586 +/- 0.003 (3 mo) to 0.581 +/- 0.002 (12 mo) to 0.569 +/- 0.003 (26 mo) at 30 degrees C. This indicated an increase in membrane fluidity with age. In membranes labeled with the 16-nitroxide stearic acid, a similar increase in membrane fluidity with age was observed. The order parameter of microsomal membranes from 3 and 26 mo rats was measured over the temperature range 10 degrees to 31 degrees C in steps of 0.9 degrees C. A plot of the log of S versus the reciprocal temperature revealed a phase transition at 24 degrees C in membranes from 26 mo rats, but no phase transition was observed in 3 mo old rats in this temperature range. The change in fluidity of the hepatic microsomal membrane with age may account for some of the observed changes in membrane-bound mixed function oxidase activities with age. PMID- 6284533 TI - A study of added GM-CSF independent granulocyte and macrophage precursors in mouse spleen infected with myeloproliferative sarcoma virus (MPSV). AB - A subpopulation of granulocyte and macrophage precursors (GM-CFUc) differentiating in the agar colony technique of Bradley and Metcalf into mature granulocytes and macrophages, without the addition of granulocyte and macrophage colony stimulating factor (GM-CSF), can be detected in MPSV infected mice. These precursors were detected 5 days after virus infection, reaching a maximal concentration of 1/10,000 spleen or bone marrow cells, 25 days after viral infection. The number of the added GM-CSF independent GM-CFUc was linearly correlated with the number of seeded MPSV hematopoietic cells. No GM-CSF producing cells could be detected in the MPSV spleen using normal bone marrow GM CFUc as responder cells. Study of the GM-CSF sensitivity of the GM precursors has demonstrated the existence of two GM-CFUc populations in the MPSV spleen: a) a GM CSF dependent population with a GM-CSF sensitivity similar to that of normal GM CFUc b) a GM-CFUc population which differentiated in the absence of detectable amount of GM-CSF and of which differentiation was not affected by the addition of progressive amounts of GM-CSF. A possible model explaining these results is proposed. PMID- 6284534 TI - Increased erythropoiesis and 2'5'-A polymerase activity in the marrow and spleen of phenylhydrazine-injected rats. AB - The rate of CFUE production in adult rats varies considerably according to whether the animals are rendered anemic by short or long term treatment with phenylhydrazine. The bone marrow responds during 2 months of phenylhydrazine injections with a progressive increase in CFUE, the spleen shows a rapid and larger rise after 3 daily injections followed by a decline to near zero, control numbers, after prolonged administration of the drug, and the liver never develops erythropoietic activity. Endogenous colonies, that is, colonies that grow in plasma clots without added erythropoietin, are most numerous in bone marrow after phenylhydrazine treatment, are always few in the spleen and are never present in cultures of liver cells. In both marrow and spleen, there is a direct correlation between the rate of erythropoiesis and 2'5'-A polymerase activity. These findings suggest that production of the enzyme may be an early event in erythroid cell differentiation. PMID- 6284535 TI - Treatment of small cell carcinoma of the lung. AB - Current strategies for treatment of small cell carcinoma of the lung are based on the concept of the disease as a systemic condition, requiring systemic treatment. Small cell carcinoma is considerably more sensitive to anticancer drugs than other bronchogenic neoplasms. The most widely used agents are cyclophosphamide, adriamycin, vincristine, CCNU, methotrexate, and VP 16-213. The superiority of combination chemotherapy over single drug treatment has been documented in prospective randomized trials. With suitable drug regimens, partial or complete remissions can be achieved in 80-90% of untreated cases, resulting in three- to five-fold prolongation of median survival. Radiotherapy may improve control of intrathoracic disease and reduce the incidence of cerebral metastases but this treatment does not increase median survival. In spite of impressive initial responses, long-term results are disappointing, with a 2-year disease-free survival rate of approximately 5 to 10%. Improvement of treatment results may be expected from the use of sequential combination chemotherapy with drugs administered at maximally tolerated doses. A high intensity of treatment calls for increased attention to supportive measures against infections and other complications. The ultimate role of surgery, brain and/or chest irradiation, and immunotherapy as adjuvants to chemotherapy remains to be defined. PMID- 6284536 TI - What goes wrong with the macrophage in silicosis? PMID- 6284537 TI - Bronchitis and exposure to man-made mineral fibres in non-smoking construction workers. AB - The purpose of the present study is to evaluate the possible association between bronchitis and exposure to man-made mineral fibres. The basis of the study has been cross-sectional data from the early 70's describing some 135,000 male Swedish construction workers. Data included information about exposure to asbestos and man-made mineral fibres, smoking habits and questions concerning symptoms of bronchitis. In non-smokers the rate-ratio of people exposed at least 3 years to non-exposed people is 2.68. In former smokers and in present smokers the corresponding values are 1.67 and 1.51 respectively. PMID- 6284538 TI - Quantal analysis of excitatory postsynaptic potentials induced in hippocampal neurons by activation of granule cells. AB - The values of quantal content (m) and quantal amplitude (q) of excitatory postsynaptic potentials (EPSPs) elicited in CA3 neurons by activation of granule cells were estimated in thin hippocampal sections maintained in vitro. For this purpose, DL-homocysteate was administered to granule cells, and trains of EPSPs that were typical for single granule cell activation were recorded from individual CA3 neurons. The amplitudes of the first and second EPSPs in each train were measured. Fron the mean and variance of the amplitude of the EPSPs, the values of q and m were calculated. The values of m and q for the first EPSPs were estimated at 8.3 and 0.28 mV, respectively, on the average. Potentiation of the second EPSPs was accompanied by a two-fold increase in the values of m without changes in the values of q. Therefore, frequency potentiation in synapses between mossy fibers and CA3 neurons may be explained by an increase in number of released quanta. Amplitudes of EPSPs were found to fluctuate in a manner described by Poisson's law. PMID- 6284539 TI - Time course and properties of late adaptation in spinal motoneurones of the cat. AB - In the spinal cord of anaesthetized cats, motoneurones of m-gastrocnemius medialis were stimulated to repetitive firing by very long-lasting steady currents injected through an intracellular microelectrode (maximum duration 4 min). In such discharges, a gradual decline in impulse frequency was found to occur during several tens of seconds. Most of this "late adaptation" occurred during the first 30 s of firing. Comparisons between the responses of different cells showed that the frequency-drop during late adaptation was strongly correlated to the impulse rate at the beginning of the discharge. For one and the same cell, late adaptation was more prominent at strong than at weaker intensities of stimulation (i.e.., at high than at lower initial firing rates). In cells capable of discharging continuously for several minutes, a semi stationary discharge rate tended to be reached after about 1 min or less. PMID- 6284540 TI - Motoneurone properties and motor fatigue. An intracellular study of gastrocnemius motoneurones of the cat. PMID- 6284541 TI - Canal-neck interaction in vestibular nuclear neurons of the cat. AB - The convergence and interaction of horizontal semicircular canal and neck proprioceptive inputs were studied in neurons of the caudal two thirds of the vestibular nuclear complex. Extracellular neuron activity was recorded under muscle relaxation and slight anesthesia in chronically prepared cats. The following stimulations were applied: horizontal rotations of (a) the whole body (labyrinth stimulation), (b) the trunk vs. the stationary head (neck stimulation), and (c) the head vs. the stationary trunk (combined labyrinth and neck stimulation). Of 152 neurons investigated, 83 (55%) showed convergence of the two inputs. In about half of these neurons, the neck input was very weak and hardly affected the labyrinthine response during head rotation. Judged from the response pattern, several of these neurons presumably were related to vestibulo oculomotor function (i.e., vestibular nystagmus). In the other half (i.e., 27% of all neurons), sensitivity of the two inputs was similar. Both labyrinthine and neck responses contained a dynamic ("velocity") component; neck responses of more than half of these neurons had, in addition, a static ("position") component. The dynamic components were either "antagonistic" or "synergistic" as to their convergence during head rotation. When applying this combined stimulation, the dynamic components summed linearly, yielding subtration in case of antagonistic convergence and addition in case of synergistic convergence. In contrast, the static components of the neck responses remained largely unchanged during head rotation. However, the static head-to-trunk deflection determined the tonic discharge level in such neurons and thus facilitated or disfacilitated the dynamic responses to superimposed labyrinth stimulation. We suggest that the two patterns of labyrinthine neck interaction observed in vestibular nuclear neurons, i.e., subtration and addition, may be involved in the postural control of the trunk and head, respectively. In contrast, interference of the neck input with vestibulo-oculomotor function appears to be almost negligible in the intact cat. PMID- 6284542 TI - Properties of ventromedial hypothalamic neurons with axons to midbrain central gray. PMID- 6284543 TI - Effects of prenatal capsaicin treatment on fetal spontaneous activity, opiate receptor binding, and acid phosphatase in the spinal cord. PMID- 6284544 TI - The guinea pig further demonstrates that rodent superior cervical ganglia differ from those of other species. PMID- 6284545 TI - A blind controlled trial of adrenocorticotropin and cerebral gangliosides in nerve regeneration in the rat. PMID- 6284546 TI - Cyclic adenosine 3', 5'-monophosphate in amygdaloid-kindled rats. PMID- 6284547 TI - Fasciola hepatica: immunisation of rats by intraperitoneal injection of adult fluke antigen in Freund's adjuvant. PMID- 6284548 TI - The effect of tetrodotoxin on the synaptic and extrasynaptic membrane in frog skeletal muscle. AB - Tetrodotoxin resistant sodium channels have been shown to operate in frog muscle membrane, which are responsible for local action potentials. These channels are located mainly in synaptic regions, and their distribution on the membrane is controlled by neurotrophic factors. PMID- 6284549 TI - Receptors for thymosin fraction V on rat thymic lymphocytes. PMID- 6284551 TI - Leader polypeptides encoded in the 5'-region of the encephalomyocarditis virus genome. PMID- 6284550 TI - [Correction of extracardiac effects on the heart as a method of treating ischemic heart disease]. PMID- 6284552 TI - The number of 5 S rRNA genes in Bacillus subtilis. PMID- 6284554 TI - Syndyphalin SD-25: a highly selective ligand for mu opiate receptors. PMID- 6284555 TI - Calmodulin-sensitive ATP-dependent calcium transport by the rat parotid endoplasmic reticulum. PMID- 6284553 TI - Expression of conserved message of poly (A) polymerase through hormonal control in wheat aleurone layers. PMID- 6284557 TI - [New stage in the study of the central effects on digestive system functions]. AB - The review included author's own studies and emphasizes the problem of peptides participating in transmission of central influences to the intramural nervous system in the digestive tract's organs, to their endocrine cells or directly to the secretory (or smooth muscle) cells. The role of endocrine apparatus of the digestive tract's organs is discussed as well as the role of the so called peptidergic fibers of the vegetative nervous system in regulation of secretory function of the stomach and pancreas. PMID- 6284559 TI - Biological activity of proviral DNA isolated from cells infected with MAV-2(0) virus. AB - The ability of DNA isolated from avian myeloblastosis-associated virus MAV-2(0) infected fibroblasts to induce the synthesis of virus upon transfection of susceptible but not resistant cells was demonstrated. The virus obtained, when inoculated into 12-day-old embryos, led to the manifestation of osteopetrosis after prolonged incubation and to the induction of nephroblastomas in some cases. Positive transfection with DNA from the non-virus producing myeloblast cell line was not detected on bone marrow cells and fibroblasts preinfected with tdB77-C virus. After transfection with DNA from virus-producing myeloblasts, the reproduction of non-transforming virus was observed. This virus did not induce myeloblastosis upon infection of chickens but induced nephroblastomas on rare occasions. PMID- 6284556 TI - Photolysis at very low temperatures of co-liganded cytochrome oxidase (cytochrome d) in oxygen-limited Escherichia coli. PMID- 6284558 TI - [Existence of neurotensin receptors in the caudate nucleus and posterior hypothalamus]. AB - Chronic experiments in dogs revealed that administration of neurotensin (3 and 10 mkg in 1.5 mkl) into the caudate nucleus altered the parameters of conditioned and unconditioned food reflexes: shortened the latency by 31% and augmented the reflexes by 56%. The microapplication of neurotensin in the same dosage into the posterior hypothalamus increased the stomach secretory function induced by histamine administration. The caudate nucleus and the hypothalamus seem to contain cells which have the receptors for neurotensin and take part in the mechanisms of conditioning and central influences on the digestive system's organs. PMID- 6284560 TI - LH releasing hormone (LHRH) - past, present and future. PMID- 6284561 TI - [Pituitary adrenal axis activity during topical treatment with 0,075 g% triamcinolone benetonide cream (author's transl)]. PMID- 6284562 TI - Mucinous (adenocystic) carcinoma of skin. PMID- 6284563 TI - Acute effects of guar gum on glucose tolerance and intestinal absorption of nutrients in rats. AB - The mechanism by which non-digestible fibres improve oral glucose tolerance is still unclear. We have studied the effects of guar gum on oral carbohydrate tolerance and intestinal absorption of nutrients in anaesthetized rats. Addition of guar to an intragastric glucose load (1 g/kg) markedly delayed the rise in plasma glucose levels when the concentration of the gum was adequate (10 mg/ml). The insulin response was somewhat less marked, but the differences were not significant. When glucose was introduced directly into the duodenum, the gum only slightly reduced the rise in glucose levels, during the first 15 min. If sucrose (1 g/kg) was infused in the duodenum, acarboseR, an alpha-glucosidase inhibitor, but not guar, slowed the rise in plasma glucose and insulin levels. Intestinal absorption was measured in a tied duodenojejunal loop. Guar decreased active transport of glucose (4 mmol/l) by approximately 20%, but had no significant effect on the passive transport of glucose (100 mmol/l), nor on the absorption of sucrose (40 mmol/l) or leucine (4 mmol/l). At the concentration which improved glucose tolerance (10 mg/ml), but not at lower concentrations, guar gum markedly slowed gastric emptying. These results suggest that guar gum improves tolerance to oral carbohydrates mainly by decreasing the rate of gastric emptying, but inhibition of intestinal absorption may also be involved in the presence of low concentrations of the sugars. PMID- 6284564 TI - Comparative non-blind trial of ceftriaxone and gentamicin in the treatment of complicated urinary tract infections. AB - Forty hospitalized patients with complicated urinary tract infections were treated with gentamicin 240 mg die i.m. or ceftriaxone, a new parenteral cephalosporin, 2 g die i.v. for 7 days. Ceftriaxone gave good clinical and bacteriological results which were better than those obtained with gentamicin, showing also an appreciable activity in cases of infection due to problematic organisms. PMID- 6284565 TI - Long-term treatment with pivmecillinam in patients with recurrent bacteriuria. AB - Thirty out-patients with chronic recurrent urinary tract infections, who had failed to respond to 10 days treatment with either pivmecillinam and/or amoxycillin, received a 3-month course of pivmecillinam at a dose of 200 mg, three times daily. Twenty-seven patients had bacteriuria due to Enterobacteriaceae, mainly Escherichia coli, sensitive to mecillinam in vitro. Pivmecillinam eradicated all the initial urinary pathogens. Reinfections occurred during treatment in three patients, who remained asymptomatic. Four subjects complained of gastro-intestinal side-effects, and therapy was withdrawn in three instances. Another three patients described unusual adverse events towards the end of the course of treatment, described as an odd sensation in the body and a desire for salt. The sensation disappeared a few days after the end of treatment. Treatment with pivmecillinam had no adverse effect on haematopoietic, hepatic or renal function. PMID- 6284567 TI - Development of cholinergic neurons of the rat retina. PMID- 6284566 TI - Bran diet for an earlier resolution of post-operative ileus. AB - The effect of a pre-operative high fibre diet on the resolution of ileus following cholecystectomy has been evaluated. The time needed to restore canalization of the gastro-intestinal tract has been compared in two random groups of patients (a total of thirty-eight) one treated with wheat bran and the other as control without the diet supplementation. The average persistence of ileus was 24 hours in the treated group and 54 hours in the control group. These results suggest that a bran-enriched diet could be an inexpensive and simple treatment to shorten the duration of ileus after abdominal surgery. PMID- 6284568 TI - Differentiation and dedifferentiation can function simultaneously and independently in the same cells in Dictyostelium discoideum. PMID- 6284569 TI - Developmental regulation of a stalk cell differentiation-inducing factor in Dictyostelium discoideum. PMID- 6284571 TI - Channel open time of acetylcholine receptors on Xenopus muscle cells in dissociated cell culture. PMID- 6284570 TI - Acetylcholine sensitivity of innervated and noninnervated Xenopus muscle cells in culture. PMID- 6284572 TI - The ascidian sperm reaction: evidence for Cl- and HCO3- involvement in acid release. PMID- 6284574 TI - Domperidone treatment in the irritable bowel syndrome. AB - Domperidone treatment did not differ significantly from placebo treatment in irritable bowel syndrome patients concurrently put on high fibre diet. No side effects attributable to the drug were encountered. This study further reinforces the necessity to have patients concurrently on a high fibre diet when testing for drug efficiency in the irritable bowel syndrome. Double-blind drug versus placebo studies should be undertaken in the irritable bowel syndrome patients who fail to respond to a high fibre diet. Dopamine receptor sites are unlikely to be primarily involved in the irritable bowel syndrome. PMID- 6284573 TI - A family of proteins accumulating in ectoderm of sea urchin embryos specified by two related cDNA clones. PMID- 6284575 TI - Binding and degradation of 125I-insulin by renal glomeruli and tubules isolated from rats. PMID- 6284577 TI - The effect of a high fibre diet on diabetic nephropathy in the db/db mouse. AB - Genetically diabetic mice (C57 BLKsJ db/db) aged 5-6 weeks were given a diet containing 20% by weight of non-nutritive bulk and compared with age matched control diabetic mice on a normal diet (fibre content 4.5%) and non-diabetic mice. The duration of study was 12 weeks. No adverse effects were observed in animals given the high fibre diet. Total food consumption was greater in mice receiving the fibre enriched diet, but their absolute caloric intake was 6% less than control diabetic mice. Both groups exhibited similar rates of growth and development. Water intake in the experimental diabetic mice was reduced and similar to that of normal non-diabetic mice. Fasting blood glucose was significantly decreased in the experimental diabetic mice at 12 weeks. Renal pathological lesions in the control diabetic mice showed glomerular mesangial expansion and deposition of immunoglobulins within the mesangium. The experimental diabetic animals exhibited significantly less renal pathology, including light and immunofluorescent lesions. It is concluded that addition of non-absorbable fibre to the diet of genetically diabetic mice improves glycaemic control and retards the development of diabetic nephropathy. PMID- 6284576 TI - Microtubules, microfilaments and insulin-secretion. PMID- 6284578 TI - Regulatory effect of glucagon on its own receptor concentrations and target-cell sensitivity in the rat. AB - To evaluate the role of glucagon on its hepatocyte receptor concentrations, groups of rats were injected with a long-acting glucagon preparation (20 [G-20], 40 [G-40] or 60 [G-60] micrograms/100 g body weight) every 8 h for 4 days. Glucagon receptors in liver plasma membranes of treated animals were decreased in number (control = 1.66 +/- 0.20 ng/0.5 mg protein versus G-20 = 1.24 +/- 0.26, G 40 = 1.03 +/- 0.26, G-60 = 0.70 +/- 0.03 ng/0.5 mg protein; p less than 0.05, less than 0.001, less than 0.001, respectively), but they were indistinguishable from receptors of control rats by other criteria including affinity and kinetics of association. Degradation of both glucagon and receptor sites did not account for differences observed in binding. Similar results were obtained with isolated hepatocytes. In relation to controls, isolated hepatocytes of treated rats had a reduced number of receptors (control = 0.70 +/- 0.05 versus G-40 = 0.47 +/- 0.04 ng/10(6) cells; p less than 0.02) proportionate to the decreased glucagon stimulated production of cyclic AMP and glucose. Four to eight hours exposure of cultured hepatocytes of nontreated rats to 4 x 10(-8) mol/l glucagon produced a decreased binding of 125I-glucagon to its receptor (p less than 0.05). In contrast, hormone exposure for shorter periods of time (0-2 h) was without effect. These results suggest (1) an inverse relationship between circulating glucagon levels and hepatocyte glucagon receptor concentration, and (2) a direct relation between receptor number and target-cell response. PMID- 6284579 TI - Antibodies to coxsackie B viruses and HLA in japanese with juvenile-onset type 1 (insulin-dependent) diabetes mellitus. PMID- 6284580 TI - [Should one treat with systemic adriamycin inoperable hepatocellular cancer in cirrhosis?]. PMID- 6284582 TI - The effects of microfilament disrupting agents on HCl secretion and ultrastructure of piglet gastric oxyntic cells. AB - A functionally responsive in vitro preparation of piglet gastric mucosa was used to investigate the involvement of microfilaments in the process of HCl secretion by oxyntic cells. A well-ordered array of microfilaments was observed in the short, stubby microvilli on the apical surface of nonsecreting oxyntic cells, as well as ii the longer microvilli of actively secreting cells. Treatment with cytochalasin B (10(-5)-10(-4) M) caused a dose-dependent inhibition of acid secretion and a concomitant gradient of morphologic alteration of oxyntic cells. Associated with slight (approximately 20%) inhibition of secretion was an initial collapse of the canalicular and glandular lumina and appearance of some pleomorphic-shaped microvilli. Maximum inhibition of secretion always produced a complete collapse of the oxyntic cell canalicular and glandular lumina, with a resultant apposition of apical surfaces. Microvilli were no longer readily distinguishable, and microfilaments were severely disorganized. Treatment with cytochalasin B (2-4 X 10(-5) M) before secretagogue stimulation also reduced the ability of the gastric mucosa to secrete acid; oxyntic cells retained the general appearance of nonsecreting cells. The correlation of disruption of microfilaments and the inhibition of acid secretion by cytochalasin B suggests an involvement of microfilaments in both the initiation and maintenance of high levels of acid secretion. PMID- 6284584 TI - Withdrawal syndromes following cessation of treatment with antihypertensive drugs. PMID- 6284581 TI - [Choleresis and cholestasis (author's transl)]. PMID- 6284586 TI - The influence of cortisol on the ACTH-producing cells in the pituitary gland of the chick embryo: an immunocytochemical study. PMID- 6284583 TI - Membranous obstruction of the inferior vena cava and hepatocellular carcinoma. PMID- 6284585 TI - Mechanisms of coupling of the beta-adrenergic receptor to adenylate cyclase--an overview. PMID- 6284587 TI - Metabolic effects of ACTH in the Tammar wallaby, Macropus eugenii: the role of the adrenal medulla. PMID- 6284588 TI - [Mapping of 2 cloned DNA fragments containing repetitive mouse genome regions, actively transcribing into pre-mRNA]. AB - Sequence organization of two cloned mouse genomic DNA fragments which are efficiently transcribed into pre-mRNA was studied. In both cases, the arrangement of sequences hybridizing to polysomal mRNA (or cDNA), double-stranded RNA (dsRNA B) isolated from pre-mRNA and oligo (dT), was established. In one fragment, the regions transcribed are represented by three different repetitive mRNA (or cDNA) hybridizing sequences interspersed into the AT-rich tract, the type B-2 sequence and the unique DNA sequence which also transcribed into mRNA. Nuclear pre-mRNA of 28S is hybridized to three repetitive mRNA hybridizing regions. The significance of such a complex array of repeated sequences inside eukaryotic transcription units and their possible role in mRNA processing and splicing is discussed. PMID- 6284589 TI - Increased stability of collagen following alloxan diabetes in Swiss mice. PMID- 6284591 TI - [Concentration of the hepatitis A virus on artificial and natural sorbents]. PMID- 6284590 TI - [Enterovirus distribution in the water of reservoirs of different climatic zones of the USSR]. PMID- 6284592 TI - [Use of x-ray spectral analysis with proton excitation for determining the joint presence of chromium, manganese, iron, nickel, cobalt, zinc, germanium, arsenic, mercury, lead and bismuth in aerosol particles]. PMID- 6284593 TI - [Properties of the soil dust released during sugar beet cultivation]. PMID- 6284594 TI - Progestin- and androgen-binding components in a human granulosa-theca cell tumor. PMID- 6284595 TI - Adenocarcinoma of the endometrium: analysis of 256 cases with carcinoma limited to the uterine corpus. Pathology review and analysis of prognostic variables. PMID- 6284596 TI - Opioid plasma levels during labour. AB - 6 healthy pregnant women, monitored with external cardiotocography, were studied. Blood samples were collected hourly throughout labour until delivery of the fetus and placenta, and again on the 5th day of puerperium. beta-Lipotropin (beta LPH) and beta-endorphin (beta EP) were determined by specific radioimmunoassays in each sample, after silicic acid plasma extraction and G-75 column chromatography. Both opioid plasma levels rise progressively during labour, reaching their highest values 1 h before delivery (beta LPH: 296.0 +/- 60.2 pg/ml; beta EP 106.2 +/- 40.6 pg/ml) as compared with values of 157.0 +/- 35.4 (beta LPH) and 57.0 +/- 7.3 pg/ml (beta EP) (mean +/- SE) at the beginning of labour. With the exception of 1 case, a significant correlation was observed between beta LPH and beta EP plasma levels. The two opioid plasma levels decrease after delivery of the placenta (272.5 +/- 59.6 and 118.2 +/- 68.4 pg/ml) and subsequently decrease further to levels of 105.6 +/- 46.4 (beta LPH) and 33.7 +/- 16.5 (beta RP) on the 5th day of puerperium. beta LPH and beta EP plasma levels showed a significant correlation throughout labour with the 'uterine contractility force/hour', which was calculated by the addition of tocographic areas of each uterine contraction at 1-hour intervals. These data demonstrate that the increase in plasma opioid concentrations during labor is directly related to the number and intensity of uterine contractions. PMID- 6284597 TI - Decreased adenosine 3'-5'-cyclic monophosphate concentration in amniotic fluid during rivanol-induced abortion. PMID- 6284598 TI - [Diagnosis of acute right ventricular infarction by noninvasive methods]. PMID- 6284599 TI - [Withdrawal of prolonged corticosteroid therapy]. PMID- 6284600 TI - [Portal hypertension in localized hepatocellular carcinoma without cirrhosis]. PMID- 6284601 TI - [Enzymes and cytochemical indices of the peripheral blood neutrophilic granulocytes in diabetics]. PMID- 6284603 TI - Lung scanning and clearance of inhaled radiogold (198Au) particles in three patients with microlithiasis. PMID- 6284602 TI - An electron microscopic study on atypical presenile dementia with numerous Lewy bodies in the cerebral cortex. AB - A report was made of an autopsied case of presenile dementia presenting the clinically characteristic symptoms of Alzheimer's disease and Pick's disease. The neuropathological findings were senile changes, namely, numerous senile plaques, neurofibrillary tangles, granulovacuolar degeneration and Pick bodies, etc., and numerous Lewy bodies in the brain stem and in the cerebral cortex. We studied mainly the ultrastructure of Lewy bodies in the cerebral cortex in this paper. The features of ultrastructural components of Lewy bodies presented types which might correspond to the various profiles of Lewy bodies observed by a light microscope. In addition, we studied the senile changes in the cerebral cortex ultrastructurally and considered that some relationship possibly existed between Lewy bodies and senile changes. PMID- 6284604 TI - [Localization of islet cell tumors using sonography, computed tomography, arteriography and selective transhepatic venous sampling for hormone assay (author's transl)]. AB - Of 29 patients examined operation revealed a malignant tumor in 9 and a benign insulinoma in 18, 2 insulinomas were not found. The problems of preoperative tumor localization were limited to small insulinomas (size 7-35 mm). Ultrasound detected all of 3 insulinomas as low echogenic structures (size 7, 8, 17 mm). Computed tomography demonstrated 4 of 5 insulinomas (size 7, 8, 15, 17 mm) due to contrast enhancement following bolus injection. Arteriography localized 12 of 18 insulinomas preoperatively and 14 of 18 retrospectively. Selective transhepatic venous sampling for insulin assay identified 7 of 8 tumors. Real-time ultrasound and dynamic CT are promising in the diagnostics of insulinomas over 7 mm and should precede arteriography. Selective transhepatic venous sampling as the last diagnostic step is a major procedure and most specific, but not always without problems in interpretation. PMID- 6284605 TI - [Technical aspects of breast sonography. Experience with an immersion scanner (Octoson) (author's transl)]. AB - The experience with the sonographic examination of 80 patients with an Octoson body unit is described. It was found that with the present technique immersion of the breast in most cases is less satisfactory than compression of the breast. The compound scan improves the information as compared with a single scan up to a point. Immersion sonography, like all other types of ultrasound examination, is at present not satisfactory as the sole method for examining the breast. As an addition to mammography and palpation, sonography is, however, already an important complementary method. PMID- 6284606 TI - [Disparate thyroid imaging with 99mTc pertechnetate and radioiodine (author's transl)]. PMID- 6284608 TI - [Adenoid-cystic carcinoma (cylindroma) of the trachea. An analysis of nine cases (author's transl)]. AB - Nine patients with an adenoid-cystic carcinoma of the trachea are describes. It is a rare tumour which grows relatively slowly; it has a tendency for early invasion of neighbouring tissues, but late distant metastases. A correct diagnosis can be made by the radiologist. The trachea should be visible on every film of the thorax and should be inspected. In particular, a diagnosis of therapy resistant asthma should direct the attention of the radiologist to the trachea. PMID- 6284607 TI - [Nephroblastoma in an adult man with multiple "osteoplastic" bone metastases]. PMID- 6284609 TI - [An analysis of 153 areas of microcalcification of malignant origin: the "triangle principle" (author's transl)]. AB - The forms and outlines of 153 clustered microcalcifications in 114 patients with histologically proven intraduct carcinomas were analysed. Seven groups could be defined; of these, a triangle principle"). The smaller groups tended to have straight margins, but the larger groups often showed wavy contours and at least one indentation at the base ("Swallow-tail phenomenon"). At the same time, the larger groups show circumscribed areas free of microcalcification, which correspond to interductal fat deposition. All these radiological appearances can be related to the anatomical structures of the lactiferous ducts. PMID- 6284610 TI - [An analysis of 136 groups of microcalcification of benign origin. How specific is the "triangle principle" (author's transl)]. AB - Amongst 107 localised groups of microcalcification of benign origin and not intraductal in localisation, there were 79 which showed round, oval or amorphous configuration, and 28 which were triangular. Of 29 intraductal areas of microcalcification, 26 showed appearances resembling an intraduct carcinoma. The "triangle principle" is therefore specific for the intraductal location of the lesion, but is unrelated to the question of malignancy. If the shape of the lesion had been used as the sole indication for biopsy, ie. had one operated only on those groups of microcalcification showing a triangular shape, it would have been possible to avoid biopsy in benign lesions in 79 patients and only three harmless intraductal lesions would have been overlooked. It is the task of the radiologist to distinguish intraductal calcification from that which is not intraductal. PMID- 6284611 TI - [Angiographic demonstration of venous tumour invasion in hepatocellular carcinomas: characteristic parallel linear vascular channels (author's transl)]. PMID- 6284612 TI - Needle biopsy and tumor staging (TNM-system). AB - Needle biopsy is helpful in classifying pulmonary neoplasms according to the TNM system. A population of 2726 patients is described in which needle biopsy differentiated between neoplasm and inflammatory disease. In the subgroup of patients with neoplasms (46%) TNM-staging was established by needle biopsy. Extrapolation of these results aided in deciding whether the diagnosed lesions were operable or inoperable. Needle biopsy is also helpful in establishing the diagnosis of oat cell carcinoma. For all practical purposes this is an inoperable lesion. The patient may such be saved unnecessary thoracotomy; an alternative treatment of surgery i.e. radiotherapy and/or chemotherapy is then available. Finally, the combination of needle biopsy with subsequent TNM-staging is useful as a prognostic device. PMID- 6284613 TI - [CT diagnosis of glomus-jugulare tumours]. AB - Glomus-jugulare tumours (chemodectomas, non-chromaffin paragangliomas) are semi malignant tumours arising from the glomus structures of the jugular bulb. They are soft tissue tumours which usually expand the jugular foramen and destroy its bony margins. Conventional methods of examination are therefore very effective. However, they only demonstrate the bone destruction and are therefore an indirect method for showing the tumour. The actual size and extent of the tumour, which are important for the surgeon or radiotherapist, cannot be demonstrated by simple x-rays. Computer tomography is very effective in the diagnosis of soft tissue lesions in the skull. CT is therefore of great importance in the diagnosis of glomus-jugulare tumours. PMID- 6284614 TI - [Arterio-venous fistula of the liver. Computer tomographic diagnosis (author's transl)]. AB - By using serial computer tomography, an arterio-venous fistula of the liver could be demonstrated in five cases. The computer tomographic findings which, previously, could only be obtained by angiography, are described. The value of the method as a diagnostic procedure is discussed and its limitations compared with angiography are documented. PMID- 6284615 TI - [Indirect breast lymphography (author's transl)]. AB - Intradermal areolar contrast injection of a new water-soluble contrast medium, Iotasul, results in regular demonstration of the latero-cranial breast lymphatics and of the subareolar lymphatic plexus. It has been possible, for the first time, to show lymph nodes with this water-soluble contrast medium, although the results are not entirely satisfactory. There have been no side effects. PMID- 6284616 TI - [The value of computer tomography in the diagnosis of local recurrences of carcinoma of the rectum (author's transl)]. AB - The tendency of rectal carcinomas to recur makes it necessary to reexamine these patients repeatedly. The tissues remaining following resection of the rectum can only be examined directly by computer tomography. The value of this procedure in the diagnosis of local recurrences has so far not been established. The results of computer tomography must therefore be correlated with already established methods, particularly clinical examination and CEA measurements. PMID- 6284618 TI - [Diagnostic charts: Cardiovascular nuclear medicine. 2. Infarct scintigraphy, radionuclide angiography, radionuclide phlebography]. PMID- 6284617 TI - [Radiologic aspects of the congenital arteriovenous malformations, Klippel Trenaunay type, and Servelle-Martorell type (author's transl)]. AB - In our study 13 patients with a Klippel-Trenaunay type and 7 patients with a Servelle-Martorell type of arteriovenous malformations were analyzed. The results demonstrate that these 2 entities can be differentiated by routine radiography. The Klippel-Trenaunay type of angiodysplasia does not need arteriography, whereas arteriography as well as phlebography are necessary in the Servelle-Martorell type of angiodysplasia, in order to show ectatic regions of the involved vessels. The differentiation between the above mentioned arteriovenous malformations and the F. P. Weber type of angiodysplasia is easy. In the F. P. Weber type of angiodysplasia a lengthening of the involved extremity, arteriovenous shunts, as well as alterations of the bone are characteristic. PMID- 6284619 TI - [Para-immunity after oral antigen administration]. AB - The term "Paramunity" summarizes all non-pathogen specific mechanisms of the defence against infections in man and animal, such as phagocytosis, interference, antibiosis, activation of the lymphopoietic cell system and of lysosomal enzymes, stimulation of humoral factors etc. Examples for the induction of a paramunity are described. After an oral or inhalatory administration of polyvalent vaccines prepared from inactivated bacteria to mice, the rate of phagocytosis of the peritoneal or alveolar macrophages increased significantly. An oral application of inactivated dyspepsia coli bacteria, caused in mice a partial protection against a challenge with a strain of Rous-sarcoma. After a tenfold oral paramunization with the polyvalent vaccine "Dodecoral" consisting of 12 heat inactivated strains of enterobacteriaceae, mice were protected against an oral infection with the parapoliomyelitis virus Col-SK. PMID- 6284621 TI - [Effect of a fiber-containing dietary formula on metabolism]. AB - 8 patients with relative weights averaging 31% above normal values as defined by Broca followed a formula diet for 2 weeks. The diet consisted of 50.7 g protein, 13.6 g fat, 91.3 g carbohydrates and also contained 14.3 g wheat bran and 3 g apple pectin as sources of roughage. The group had a mean weight loss of 5.9 kg (421 g/day). Blood biochemistry estimations showed a decrease in cholesterol from 197 +/- 13 to 166 +/- 13 mg/day (p less than 0.01). There were no disorders of water and electrolyte balance. Transaminases remained within the normal range. No increase in leucocytes and erythrocyte sedimentation rate was observed. Physical efficiency of the patients was tested intermittently on days 7 and 13 by means of muscular training lasting 180 minutes. This did not lead to additional disorders of water and electrolyte metabolism except for a transient increase in magnesium excretion during muscular activity. In the second week the mean weight of the stools was 79 +/- 11 g/day. Because of the adequate supply of dietary fibre no constipation was noted. PMID- 6284622 TI - Dietary habits and colonic carcinoma. PMID- 6284620 TI - [Early summer meningoencephalitis]. PMID- 6284623 TI - Lymphocyte transformation to specific antigens associated with Crohn's disease. AB - Specific lymphocyte transformation to three micro-organisms that have been implicated in Crohn's disease was performed. These were Pseudomonas maltophilia, Mycobacterium kansasii and Rotavirus. No significant differences were observed for any of the antigens tested. Simultaneous testing of nonspecific stimulation by PHA also showed no difference between patients and controls. These results do not support an aetiological role for these organisms mediated by a purely cellular mechanism. PMID- 6284624 TI - C-peptide concentration in pancreatic juice obtained during endoscopic pancreatography in patients with and without insulinoma. AB - ERCP was performed in three patients with insulinoma. One had a large malignant tumor, while the remaining two had small tumours. In two of these patients pancreatic juice was collected for C-peptide determination. Pancreatography was performed and pancreatic juice was obtained in seven other subjects comprising: five control subjects and two patients in whom insulinoma was suspected because of symptoms suggestive of hypoglycaemia. Pancreatography was normal in all subjects except the patient with a large insulinoma in whom an obstruction of the main pancreatic duct was found. The maximal C-peptide concentrations in pancreatic juice of patients with insulinoma were found to be several-fold higher than in the control subjects and in one of the patients in whom insulinoma was suspected but unproven. The remaining patient with suspected insulinoma had a maximal C-peptide concentration comparable with those found in patients with proven insulinoma. Thus remarkable differences in maximal C-peptide concentrations obtained in patients with and without insulinoma were found. However, the clinical significance of the findings needs further evaluation. The value of ERP in patients with suspected insulinoma may be twofold: an obstruction of the main pancreatic duct may indicate a large, hardly resectable tumour; in patients in whom the duct is unaffected the relation between the tumour as visualized by angiography, and the duct, is of value for the surgeon when planning the operation. PMID- 6284625 TI - Hepatitis Be antigen and antibody in chronic liver diseases and hepatocellular carcinoma. AB - The presence of hepatitis Be antigen (HBeAg) and antibody (anti-HBe) was investigated by immunodiffusion in 144 patients with chronic liver disease, and 129 with hepatocellular carcinoma (HCC). Most of the patients were HBsAg positive. In 62 patients with chronic active viral hepatitis B, 17 (27%) were positive for HBeAg and 25 (40%) for anti-HBe. HBeAg and anti-HBe were not related to the degree of histological activity or serum alanine aminotransferase activities, but were related more frequently to higher HBsAg titer and younger age; whereas anti-HBe generally correlated in an opposite manner. Two patients seroconverted from HBeAg to anti-HBe in 13 and 20 months respectively. HBs antigenemia was not eliminated in either HBeAg or anti-HBe positive patients. The prolonged interval in seroconversion and an age-related declining frequency of HBeAg, accompanied by a reciprocal increase in anti-HBe in chronic infection, suggest anti-HBe as a chronologic indicator in HBs antigenemia in chronic HBsAg carriage. In HCC, regardless of coexisting cirrhosis, a predominant frequency of anti-HBe (62%) was found as in cirrhosis (54%), suggesting longstanding HBsAg carriage in these patients. PMID- 6284626 TI - Post mortem changes in drug-metabolizing enzymes of rat liver and human liver. AB - The aim of this investigation was to obtain information on the time-dependent decrease of the drug-metabolizing system in autolysing rat liver, and also in human cadaver liver. Rat liver, divided into three parts, was tested immediately after removal and 6 and 12 hrs later. Parameters investigated were: microsomal protein, cytochrome P-450, NADPH cytochrome C reductase, glucose-6-phosphatase, aminopyrine-N-demethylation and aniline-p-hydroxylation. In human liver, samples taken from 0.5 up to 3.5 hrs after death, microsomal protein cytochrome P-450, NADPH cytochrome C reductase and phospholipids were tested. Nearly all parameters based on microsomal protein decrease during autolysis, but by different amounts. Interestingly, the cytochrome P-450 content of patients with signs of shock 12 hrs before death is significantly lower than in patients without shock. PMID- 6284627 TI - Alpha-2 adrenergic antilipolytic effect in dog fat cells: incidence of obesity and adipose tissue localization. AB - The main intention of this study was to characterize the alpha-adrenoceptor responsible for the inhibition of lipolysis in dog fat cell and to define circumstances that may be associated to a modification of the alpha-mediated antilipolytic effect. Isolated fat cells from omental and subcutaneous adipose tissue from normal and obese dogs were used. Basal and theophylline stimulated lipolysis was studied in the presence of selected alpha-adrenergic agonists and antagonists. The antilipolytic effect of catecholamines is mediated by alpha 2 type adrenoceptors in dog fat cell. The alpha-adrenergic responsiveness is enhanced (or unmasked) in large fat cells of obese dogs and depends on the site from which the adipose tissue sample is taken. The alpha-response is stronger in subcutaneous than in omental adipocytes. In conclusion, the weakened lipolytic responsiveness to epinephrine of obese dog fat cells seems related to an increased alpha-adrenergic response rather than a decreased beta-lipolytic effect. Obesity is a circumstance characterized in the dog fat cell by a modification of the balance between alpha-2 and beta receptors. PMID- 6284628 TI - Catecholamine and thyroid hormone influence on brown fat Na+, K+-ATPase activity and thermogenesis in the rat. AB - Resting oxygen consumption (VO2) before and after injection of noradrenaline (NA), and plasma triiodothyronine levels were elevated in hyperthyroid and hyperphagic cafeteria fed rats, but were reduced in 4d-fasted and hypothyroid animals compared to controls. Refeeding fasted rats with a single carbohydrate meal caused all of these parameters to increase towards control levels. In vivo turnover, and in vitro release of NA brown adipose tissue (BAT) was elevated in cafeteria fed rats but remained unaltered in other groups and levels and uptake of NA in BAT were similar for all rats. Basal and NA stimulated Na+,K+-ATPase activity in BAT was increased in cafeteria and hyperthyroid rats and reduced in fasted and hypothyroid animals compared to control and refed groups. A highly significant correlation (r = 0.977), (P less than 0.001), found between the in vitro activity of this enzyme and resting VO2 in all rats, indicates that BAT Na+,K+-ATPase may be involved in the thermogenic responses to diet, catecholamines and thyroid hormones. PMID- 6284629 TI - Inhibition of bovine brain cyclic nucleotide phosphodiesterase by a proteinaceous factor from Escherichia coli. AB - An inhibitory factor for Ca2+ and calmodulin-dependent cyclic nucleotide phosphodiesterase of bovine brain was present in the soluble fraction of Escherichia coli. The factor was heat-stable but trypsin sensitive. The activity of brain phosphodiesterase supported by Ca2+ and calmodulin was inhibited by the factor in a dose dependent manner, but the basal activity was not affected. The inhibition of phosphodiesterase induced by the factor could be abolished by adding large amount of calmodulin, but not by increasing concentration of Ca2+. It was suggested that the factor interacted with calmodulin and thereby inhibited the phosphodiesterase. The factor may be a calmodulin-binding protein in E. coli. PMID- 6284630 TI - Hepatocellular carcinoma in women: probable lack of etiologic association with oral contraceptive steroids. AB - To investigate the possibility of an association between oral contraceptive steroids (CS) and hepatocellular carcinoma (HCC), we reviewed 128 cases of HCC in women collected between 1953 and 1980. There were 48 cases under the age of 40, and 13 of these (27%) had used CS. However, 62% of HCC associated with CS and 58% of HCC in women under 40 not using CS were classified histologically as "fibrolamellar" carcinoma. This subtype of HCC occurs predominantly in young people, both male and female. The apparent increase in HCC in young women can be explained by the presence of cases of fibrolamellar carcinoma in this age group, an the apparent association with CS is probably coincidental. PMID- 6284632 TI - Signet-ring cell adenocarcinoma of rectum: a histological, histochemical and electron microscopic study. AB - A signet-ring cell adenocarcinoma of rectum removed from a 71-year-old woman was examined by light and electron microscopy. Most of the mucin in the signet-ring cells was of the neutral type, although acidic, mostly non-sulphated, mucin was also present. Three other types of neoplastic cells. The similarities between this tumour and tumours described in the stomach, appendix and breast care discussed. PMID- 6284631 TI - Dendritic reticulum cell sarcoma? Four cases of a lymphoma probably derived from dendritic reticulum cells of the follicular compartment. AB - This paper describes the histological picture of four tumours of the follicular compartment of the lymph node, in which the proliferating cell appeared to be the dendritic reticulum cell (DRC). This assumption was based on the results of light microscopical, ultrastructural, immunological, and enzymehistochemical investigations. The tumour cells resembled DRC's closely in (1) the striking pattern of interdigitations and occasional tight junction-like contacts between the neoplastic cells on electron microscopical analysis; (2) presence of receptors for the activated third component of complement on the membrane of the cells; (3) absence of monoclonal immunoglobulins and T-cell antigen on the surface and of lysozyme, alpha 1-antitrypsin or alpha 1-antichymotrypsin in the cytoplasm of the neoplastic cells. Moreover, (4) the tumour cells showed moderate alpha-naphtyl acetate esterase, weak to absent acid phosphatase and (with one exception) strong 5-nucleotidase activity. Furthermore, (5) the neoplastic cells expressed Ia-like antigens on the surface in all four cases. The relation with follicle centre cell lymphomas, the differential diagnosis and clinical data are discussed. PMID- 6284633 TI - Primary malignant fibrous histiocytoma of the spleen: an ultrastructural study. AB - A primary malignant fibrous histiocytoma of the spleen was studied by light and electron microscopy and is believed to be the first reported. The neoplasm was predominantly fibroblastic, with a characteristic storiform pattern, and included histiocyte-like cells, giant and foam cells confirmed by ultrastructural studies. Additionally, undifferentiated cells, intermediate cells and myofibroblasts were seen. The differential diagnosis from other sarcomas and the histogenesis of this tumour are discussed. PMID- 6284635 TI - Cytolytic T lymphocyte clones. PMID- 6284634 TI - Search for causative connections between neurophysiological and psychological phenomena in the investigation of perception. PMID- 6284637 TI - Suppression of experimental allergic thyroiditis in rats treated with lithium chloride. PMID- 6284636 TI - Studies on the interrelation of resistance and immunity in a mouse model system of herpes-simplex type 2 infection. AB - Intraperitoneal (i.p.) vaccination of mice with attenuated herpes simplex virus type 2 (HSV 2) induced solid protection to i.p. infection with pathogenic virus within two days. Protection was non-virus-specific until day four after sensitization but increased in specificity thereafter. Normal mice could be protected by adoptively transferred spleen cells, serum, and peritoneal fluid from donors vaccinated seven days before. Virus-specific effector cells induced in the spleen by in vivo i.p. sensitization with either live, pathogenic, or attenuated virus and tested in a cytotoxicity assay were exclusively B lymphocytes. No functional B cells, but natural killer (NK) cells, could be detected in the unseparated peritoneal exudate cell (PEC) population. Ability to generate HSV 2 specific antibody responses did not correlate with natural resistance. PMID- 6284639 TI - Enhancement of mitogen-induced lymphocyte proliferation after preincubation is due to altered sensitivity to prostaglandins. AB - Twenty-four hour preincubation at 37 degrees enhances the mitogen-induced DNA synthesis of human lymphocytes. PGE1, given simultaneously with Con A at the start of lymphocyte culture, inhibits the DNA synthesis. After 24 h preincubation of cells, PGE1 fails to decrease the DNA synthesis. Similarly, preincubation abolished the effect of indomethacin increasing DNA synthesis of freshly separated lymphocytes. The intracellular cAMP level of human mononuclear leukocytes rapidly decreases during in vitro incubation at 37 degrees C. PGE1 elevates the intracellular cAMP of freshly separated lymphocytes to 45 times of its starting level. After 24 h preincubation of cells at 37 degrees C, PGE1 elevates cAMP to a lesser extent. This change of PGE1 action may explain the fact that the effect of exogenous PGE1 and endogenous prostaglandins (the production of which can be inhibited by indomethacin) diminishes after incubation of lymphocytes. It is very likely that the change of the effect of prostaglandins produced in lymphocyte cultures and the spontaneous decrease of intracellular cAMP level explains the enhancement of mitogen-induced lymphocyte proliferation after 24 h preincubation at 37 degrees C. PMID- 6284638 TI - Macrophage aggregation factor: some properties. AB - The lymphokine activity, macrophage aggregation factor (MAgF) has been investigated further. Activity was consistently found in 24 hr test, but not control, spleen cell culture supernatants. This was higher after dialysis against water, than in the original culture supernatants. MAgF was heat-stable, inactivated by alpha-chymotrypsin, partially inactivated by trypsin and not affected by neuraminidase. Activity was recovered from the supernatant after protein precipitation with 1 M perchloric acid, leading to a modest purification. Activity was only marginally reduced after treatment with periodate, and was not absorbed by Concanavalin A-Sepharose. Polyacrylamide gel electrophoresis showed that MAgF migrated cathodally to albumin. Aggregation, as measured in a batch centrifugation assay, was an expression both of cell-substrate and cell--cell adhesion. PMID- 6284640 TI - Oxidant-mediated damage of Leishmania donovani promastigotes. AB - Dissemination of Leishmania within the host is related to parasites undergoing unchecked proliferation. We therefore studied the effects of oxidant generating systems on promastigote multiplication by (i) direct determinations of organism proliferation and (ii) the incorporation of [3H]uracil into promastigote nucleoprotein. These two parameters correlated closely as measures of organism replication as demonstrated by parallel suppression of them by the protein synthesis inhibitors puromycin and cycloheximide and the nucleic acid synthesis inhibitors actinomycin D and mitomycin C. Promastigotes showed dose-related susceptibility to reagent and generated hydrogen peroxide (H2O2) as reflected in quantitatively similar decreases in multiplication and [3H]uracil incorporation. These effects were specific for H2O2 as catalase abrogated the dimunition in multiplication. The generation of superoxide anion by acetaldehyde-xanthine oxidase (10 mU/ml) did not alter promastigote replication or nucleoprotein synthesis. These results indicate that Leishmania donovani promastigotes are damaged by H2O2 and that the incorporation of [3H]uracil into promastigote nucleoprotein may be useful for studying the interaction of this parasite with host effector cells. PMID- 6284641 TI - Infection enhancement of dengue type 2 virus in the U-937 human monocyte cell line by antibodies to flavivirus cross-reactive determinants. AB - Dengue type 2 virus replication was detected in the U-937 human monocyte cell line when the virus inoculum and the culture medium contained flavivirus antibodies diluted beyond their neutralizing titers. This was in marked contrast to yellow fever virus, which replicated very well in the absence of antibodies; however, 10-fold-higher yields of yellow fever virus could be obtained in the presence of flavivirus antibodies. These infection-enhancing antibodies were obtained from either a dengue type 2 human antiserum or reference hyperimmune obtained from either a dengue type 2 human antiserum or reference hyperimmune mouse ascitic fluid. The infection enhancement phenomenon, previously shown to be due to infection of Fc receptor-bearing cells with virus-antibody complexes, was completely blocked by preincubation of the cells with aggregated gamma globulin. The blocking results suggested an Fc receptor-mediated infection of the U-937 cells as well. A panel of monoclonal antibodies, previously characterized as either virus type specific or flavivirus cross-reactive and with mouse immunoglobulin subclasses G1 and G2a in both categories, were tested for their infection enhancement characteristics. A type-specific neutralizing monoclonal antibody preparation that was diluted beyond its neutralization titer did not cause infection enhancement, nor did low-level neutralizing monoclonal antibodies that were dengue serotype specific by the hemagglutination inhibition test. Only flavivirus cross-reactive monoclonal antibodies caused infection enhancement, irrespective of whether the immunoglobulins were G1 or G2a. These cross-reactive flavivirus determinants may reside at the tips of the glycoprotein projections on the virus particles, enabling the Fc ends of the cross-reactive antibodies attached to these determinants to interact with Fc receptors on susceptible cells. PMID- 6284642 TI - Natural cell-mediated cytotoxicity of bovine mononuclear cells against virus infected cells. AB - The ability of mononuclear cells (MC) from peripheral blood of normal cattle to lyse a variety of cells was tested in a 51Cr-release microcytotoxicity assay. Several types of bovine cells infected with parainfluenza 3 virus (PI3V) were susceptible to natural cytotoxicity. Bovine cells infected with infectious bovine rhinotracheitis virus or noncytopathogenic bovine viral diarrhea virus, uninfected bovine cells, and human cell lines MOLT-3, HSB-2, K562, and U-937 were not susceptible. The period of time that target cells need to be infected with PI3V to achieve maximal cytotoxicity was determined. Target cells were infected with PI3V and MC, added 1 h later. After the addition of effector cells, significant levels of cytotoxicity were recorded at 17 h. Maximal cytotoxicity occurred 22 to 24 h postinfection. To define the optimal time that MC must be present, cells were infected with PI3V for a total of 24 h, and MC were left in contact with target cells for various time intervals. Maximal cytotoxicity was recorded when effector cells were present for 20 h, suggesting that a period of activation was needed to stimulate effector cell function. Removal of adherent mononuclear cells on Sephadex G-10 columns did not reduce the low level of cytotoxicity against uninfected target cells, but markedly reduced the level of cytotoxicity against PI3V-infected cells. The effector cell was nonphagocytic and nonadherent. These characteristics and the fact that target cell lysis was independent of genetic restriction indicate that effector cells are similar to natural killer cells described in other species. PMID- 6284643 TI - In vivo induction of anti-herpes simplex virus immune response by type 1 antigens and lipid A incorporated into liposomes. AB - To establish the requirements for a potential subunit vaccine against herpes simplex virus (HSV), we analyzed the effects on immunogenicity of incorporating detergent-extracted glycoprotein-enriched HSV type 1 (HSV-1) antigens into liposomes alone or with the adjuvant lipid A. Incorporating HSV-1 antigens into liposomes enhanced their immunogenicity for antibody production as detected by radioimmunoassay. Antibody levels to free and liposome-bound antigens were enhanced by administering lipid A as an adjuvant. The maximum immunogenic effect was obtained by incorporating lipid A into liposomes containing the HSV-1 proteins. Such liposomes induced secondary antibody responses higher than those engendered by virus infection. Whereas infectious virus induced cell-mediated immunity detectable by the delayed-type hypersensitivity reactions and cytotoxic T lymphocyte production, none of the liposome preparations induced cell-mediated immunity. PMID- 6284644 TI - Role of macrophage oxidative metabolism in resistance to vesicular stomatitis virus infection. AB - The role of oxygen metabolites in mediating virucidal activity was studied in two cloned macrophage-like cell lines. The parental cell line, J774.16, upon appropriate stimulation with either phorbol myristate acetate (PMA) or aggregated immunoglobulin, is induced to oxidize glucose via the hexose monophosphate shunt and produce O2- and H2O2. A variant derived from it, clone C3C, is defective in oxidative metabolism and cannot be stimulated to produce O2- or H2O2. Significant differences in yields of vesicular stomatitis virus (VSV) between stimulated clone 16 cells and unstimulated cells could be obtained only when low multiplicities were used for infection. Under the same conditions, PMA stimulation of the variant clone C3C produced no reduction in yields. The effect of PMA on virus yields in clone 16 was short-lived and dose dependent. PMA stimulation of either cell line had no effect on the number of infectious centers, suggesting that the antiviral effect was likely to be an extracellular, rather than an intracellular, one. Using glucose oxidase plus aglucose to generate H2O2 in solution, we observed that H2O2 alone is capable of killing limited amounts of VSV. The inactivation of VSV, both by H2O2 in solution and by activated clone 16 cells, could be inhibited by catalase. We conclude that intracellular resistance to VSV is primarily mediated through nonoxidative mechanisms, since activated macrophages can kill only a limited number of infectious virus particles extracellularly by means of secreted H2O2. PMID- 6284646 TI - Electrophoretic analysis of polypeptides immune precipitated from cytomegalovirus infected cell extracts by human sera. AB - Serodiagnosis of cytomegalovirus (CMV) infection by complement fixation tests depends on showing a fourfold rise in antibody titer from acute- to convalescent phase sera. Freeze-thaw and glycine-extracted, infected cell culture antigens used for these tests give markedly different titers in reactions with the same sera. In this study, we characterized the CMV-infected cell polypeptides contained in freeze-thaw and glycine-extracted antigens and identified the proteins precipitated by 23 pairs of human acute and convalescent sera. Our results were as follows. First, freeze-thaw and glycine-extracted antigens prepared from infected cells radiolabeled with [35S]methionine and subjected to electrophoresis in sodium dodecyl sulfate-polyacrylamide gels yielded similar patterns, and the bulk of the label was contained in late structural proteins and glycoproteins. Glycine-extracted preparations contained a greater proportion of soluble 66,000- and 50,000-molecular-weight proteins than did freeze-thaw antigens. Second, convalescent sera precipitated proteins migrating with apparent molecular weights of 150,000, 130,000, 110,000, 96,000, 74,000, 66,000, 50,000, 34,000, 32,000, and 25,000. Of these the 130,000-, 110,000-, 96,000-, 66,000-, 50,000-, and 25,000-molecular-weight proteins comigrated with glucosamine-labeled polypeptides. Both immunoglobulin G and M antibodies in human sera precipitated these proteins from CMV-infected cell preparations. Implications of the results for serodiagnosis of CMV infections are discussed. PMID- 6284645 TI - Effects of antiviral agents on murine cytomegalovirus-induced macrophage dysfunction. AB - Infection of murine peritoneal macrophages with murine cytomegalovirus (MCMV) led to disruption of phagocytosis. This alteration of cellular behavior appeared to be an early event in viral replication appearing 24 to 36 h before virus production and 84 to 108 h before cell death. The effects of a variety of antiviral agents on both MCMV replication and MCMV-induced depression of phagocytosis were evaluated in vitro. Although all compounds thought to act by preventing viral DNA replication inhibited MCMV replication in macrophages, none prevented expression of virus-induced alteration of phagocytosis. Cycloheximide at 1 microM blocked viral replication and viral antigen expression and prevented depression of phagocytic activity. PMID- 6284648 TI - Discovery of cytomegalovirus in pericarditis fluid from a patient with neoplasm. PMID- 6284647 TI - Effect of lymphocytosis-promoting factor from Bordetella pertussis on cerebellar cyclic GMP levels. AB - Dermonecrotic toxin (DNT), lipopolysaccharide (LPS), and lymphocytosis-promoting factor (LPF) were isolated from Bordetella pertussis and tested for neuroactivity. When injected intraperitoneally into rats, a dose of 0.13 mg of LPF per kg elevated the cyclic GMP level in cerebellum by approximately 70%, whereas DNT (0.5 mg/kg) and LPS (1.5 mg/kg) were without effect. This action of LPF on the central nervous system was dose dependent and did not require the administration of any additional agent, such as histamine. PMID- 6284649 TI - Elimination of bacteria in biliary tract infections during ceftizoxime therapy. AB - A transpapillary indwelling catheter was inserted to prevent stone impaction in six female patients who were suffering from choledocholithiasis. The bile withdrawn via the catheter was infected on six occasions with Escherichia coli. In one of these cases Klebsiella sp. and in another Salmonella sp. were also identified. All bacteria were sensitive to ceftizoxime (the MIC was between 0.007 and 0.06 mg/l). The bacterial counts in the bile were determined before and during treatment by means of membrane filtration. In all six cases there was a rapid decline in the colony count. The concentration of ceftizoxime in bile samples was several times higher than the MIC of ceftizoxime for the corresponding pathogens. Overall, the therapeutic results with ceftizoxime were good. Three of eight pathogens were eliminated from the bile within eight to 24 hours. In one case a change of pathogen was seen after 24 hours. Forty-eight hours after beginning treatment, four of eight pathogens had been eliminated from the bile. After 72 hours the colony count in six patients was less than 10 pathogens/ml. In two patients a change of pathogen occurred; in one patient treatment had to be stopped after the first injection because of urticaria. PMID- 6284650 TI - [Penetration of ceftizoxime (FK 749) into the cerebrospinal fluid (author's transl)]. AB - After the cytology and the chemistry of the CSF had returned to normal, the concentration of ceftizoxime was determined in the CSF and in the serum of 27 patients who had suffered from purulent or serous meningitis. The patients were divided into four groups. The concentration in the CSF was mean = 0.87 mg/l two hours after a bolus injection of 2 g ceftizoxime. After an infusion of 2 g ceftizoxime over 30 minutes, the concentration in the CSF was mean = 0.32 mg/l one hour and mean = 0.99 mg/l two hours after the infusion was started. The highest concentrations in the CSF were obtained six hours after a bolus injection of 2 g ceftizoxime. The peak value was 4.3 mg/l and the mean 1.78 mg/l. Based on these pharmacokinetic data, additional injections could have a cumulative effect on the concentration of ceftizoxime in the CSF. With the exception of Staphylococcus sp., Pseudomonas sp. and some anaerobes, it can be expected that ceftizoxime will be effective against other organisms causing meningitis. The results of clinical applications remain to be seen. PMID- 6284651 TI - The combination of pivmecillinam and pivampicillin compared to co-trimoxazole in the treatment of enteric fever. AB - Typhoid fever is an infectious disease with multisystem involvement and is commonly seen in the tropics. Twelve patients with acute typhoid fever were successfully treated with a fixed dose combination of pivmecillinam and pivampicillin. The treatment results were compared to those obtained from the treatment of ten other patients with co-trimoxazole, which is the routine treatment of our Department. The two forms of treatment appeared to be equally effective, suggesting that the combination mecillinam/ampicillin may represent a valuable alternative to the antityphoidal drugs currently available. Eleven patients were infected with strains resistant in vitro to either ampicillin, chloramphenicol, or both. All clinical isolates were sensitive to co-trimoxazole and to the combination of mecillinam and ampicillin. MIC values for the combination ranged from 0.16 to 2.5 mg/l. No side-effects were recorded. PMID- 6284652 TI - Mecillinam alone and in combination with ampicillin or moxalactam in experimental Escherichia coli meningitis. AB - The activity of mecillinam, ampicillin and moxalactam alone and in combination was determined in a lapin meningitis model and a mouse meningitis model against two Escherichia coli strains isolated from infants with meningitis. Both strains were highly susceptible in vitro to the antibiotics, and responded well in systemic mouse protection tests (PD50 less than 4 mg/kg). Continuous infusion of mecillinam in the lapin model over nine hours was effective in sterilizing the CSF of three of four animals infected with one strain. This prompt bacteriologic response to mecillinam alone precluded the possibility of constant infusion administration for synergy studies. Therefore, single dose administration was used to demonstrate the synergistic potential of mecillinam with ampicillin in the lapin meningitis model against the E. coli Kl # 2 strain. The combination of moxalactam and mecillinam was synergistic against the E. coli Kl # 2 strain in the mouse meningitis model. The synergistic potential of these combinations could not be reliably predicted by in vitro tests, time kill curves or systemic mouse protection tests. PMID- 6284653 TI - The cooperation of cefotaxime and desacetyl-cefotaxime with respect to antibacterial activity and beta-lactamase stability. AB - Desacetyl-cefotaxime, the main metabolite of cefotaxime, possesses a broad antibacterial spectrum. Its activity is lower than that of cefotaxime, but significantly surpasses that of cefazolin against gram-negative bacteria. The beta-lactamase stability of desacetyl-cefotaxime is higher than that of the parent compound. This beta-lactamase stability might be responsible for the synergistic effects of cefotaxime and desacetyl-cefotaxime occasionally seen in in vitro combination studies. PMID- 6284654 TI - Hepatectomy in children. PMID- 6284655 TI - Factors mediating the binding of immune complexes to cryostat sections of psoriatic lesions. AB - Sheep erythrocytes sensitized with rabbit IgG, or complexes of horseradish peroxidase (HRP) and rabbit IgG antibodies to HRP bound to cryostat sections of psoriatic and other skin lesions (lichen planus, discoid lupus erythematosus, sarcoidosis, and mycosis fungoides). Immune complexes with reduced and alkylated IgG (RA-IgG) reacted with preparations of soluble rheumatoid factors (RF) and with sections of RF-coated sheep erythrocytes equally well as did untreated IgG. However, RA-IgG did not react with sections containing FcR-positive cells. These complexes bound either weakly or not at all to skin sections. IgG inhibited the binding of immune complexes to all skin control sections, while RA-IgG only inhibited the binding of complexes to sections of RF-coated cells. Anti-Clq did not inhibit the binding of immune complexes to any section. Periodic acid and formaldehyde abolished FcR activity, whereas neither the activity of soluble RF nor of RF in sections was affected. Only sections of lesional skin from patients with highly active psoriasis and sarcoidosis showed some binding of immune complexes after treatment with periodic acid or formaldehyde. Apparently, the binding of immune complexes in vitro to sections of psoriatic and other skin lesions is mainly mediated by FcR. PMID- 6284656 TI - A monoclonal antibody (B72.3) defines patterns of distribution of a novel tumor associated antigen in human mammary carcinoma cell populations. AB - We report here both the range and patterns of reactivity of an IgG1 monoclonal antibody, B72.3, prepared against human, metastatic mammary carcinoma cells. When the avidin-biotin complex (ABC) immunoperoxidase technique was used on tissue sections, monoclonal B72.3 reacted with 19 of 41 (46%) primary mammary carcinomas and 13 of 21 (62%) metastatic lesions, either in axillary lymph nodes or at distal sites. Variable concentrations of antigen, recognized by B72.3, were observed among mammary tumors, as well as among different cell populations of a given tumor mass. Several patterns of antigen distribution were observed: membrane, diffuse cytoplasmic, focal and marginal. No reactivity was observed to normal mammary epithelium, stroma, or lymphocytes of the breast, nor to any cell types in a variety of other normal human tissues, melanomas, and sarcomas. Reactivity with all of four colon carcinomas was also observed. Assay of serial sections of mammary carcinomas with B72.3 and a monoclonal antibody directed against carcinoembryonic antigen demonstrated that these antigens were both distinct and non-coordinately expressed. PMID- 6284658 TI - Separation of high mammary tumor incidence from high hepatoma incidence in backcross mice during segregation of the viable yellow gene. PMID- 6284659 TI - Suppression of spontaneous mammary tumorigenesis despite Mtv-1 gene expression in hybrid and backcross C3H-AvyfB X C3H/Sm mice. PMID- 6284657 TI - Tumorigenicity of human BK papovavirus plaque isolates, wild-type and plaque morphology mutant, in hamsters. AB - The polyoncogenic prototype human papovavirus BKV (Gardner's strain) produced clear, small and large plaques. Two small-plaque isolates (wt-500 and wt-502), as well as a large-plaque-forming isolate (wt-501), induced frequent brain tumors and occasional osteosarcomas (but no insulinomas) in hamsters. The large-plaque former (wt 501) was more tumorigenic than the two small-plaque mutant (pm-525), which had been rescued from hamster tumor cell lines and have small deletion near the origin of DNA replication, induced frequent brain tumors and insulinomas. Mutant 522 was about five times more tumorigenic than wt-501. These results support the hypothesis that the polyoncogenicity of prototype BKV is accounted for by the presence of BKV mutants. PMID- 6284660 TI - Organotropism of the lymphoproliferative disease virus (LPDV) of turkeys. AB - Turkey poults were inoculated with lymphoproliferative disease virus (LPDV) of turkey, and the organotropism of the inoculated virus was determined from the kinetics of virus expression and tumor formation. Molecular hybridization experiments, conducted to determine the level of viral RNA expression in the various organs of infected birds, established lymphoid tissues, including bone marrow, as the target for LPDV infection. Of these, bone marrow was the first to be infected but subsequently virus replication extended to the thymus, the spleen and the bursa Of Fabricius. The low level of LPDV expression in non-target organs probably stemmed from infiltration of infected lymphatic cells. The fact that the main organs for virus replication were not invariably the ones most clinically involved suggests that the target cells for virus infection need not necessarily be the targets for virus oncogenicity. A possible mode for LP[DV infection and transformation is proposed. PMID- 6284661 TI - Diltiazem: effects on exercise performance in patients with coronary artery disease. AB - We evaluated a new slow-channel calcium-blocking agent, diltiazem hydrochloride, in 10 patients with documented fixed coronary artery disease who had reproducible angina and greater than or equal to 0.1 mV ST segment depression on ECG treadmill exercise testing after 1 week of single-blind placebo administration. Subsequently, over the next 6 weeks, either diltiazem (30 mg/tablet) or placebo were administered for 1 week each in a randomized double-blind triple crossover design, as one tablet four times a day, two tablets three times a day or two tablets four times a day, for a total diltiazem dose of 120, 180 and 240 mg/day, respectively. Treadmill (electrocardiogram) exercise testing was performed at the end of each week. Only diltiazem at 240 mg/day increased significantly the time to angina pectoris (P less than 0.05), time to ST segment depression (P = 0.01), time to maximal exercise (P less than 0.02), and heart rate at maximal exercise (P less than 0.05) without effecting significantly the maximal rate-pressure product compared to the corresponding placebo values. In addition, using only the diltiazem data, a significant high dose response was observed for time to angina (P less than 0.05), ST segment depression (P less than 0.005), and maximal exercise (P less than 0.025). No adverse reactions were reported during the study. Therefore, we conclude that 240 mg/day of diltiazem improves significantly exercise performance in patients with angina pectoris due to fixed coronary artery disease and adverse effects, rarely, if ever, occur at this dosage. PMID- 6284662 TI - Calcium release from membrane-bound calcium pool induced by ACTH. AB - We examined the effect of ACTH on the binding of Ca2+ to the plasma membrane of rat fat cells. Synthetic I-24 ACTH reduced the binding of 45Ca to the fat cell membrane dose-dependently. Whereas verapamil significantly increased binding, Ca ionophore A23187 significantly reduced it. These results suggest that ACTH induced the promoted release of Ca2+ from the plasma membrane-bound Ca pool into cytosol. ACTH-induced binding of 45Ca to the fat cell membrane occurred at at least two binding sites, i.e., high and low affinity binding sites. Epinephrine did not significantly influence 45Ca binding, showing an evident difference in the Ca2+ dependence of the lipolytic actions of ACTH and epinephrine. PMID- 6284663 TI - A review of recent contributions on biologically active products of arachidonate conversion. AB - Leukotrienes (LTs) C4, D4 and E4, the recognized components of slow reacting substance of anaphylaxis (SRS-A), have previously been shown to have contractile activities for guinea pig pulmonary and ileal smooth muscles; LTB4 has been shown to possess chemotactic activity for neutrophils in vitro. Based on data obtained by the use of structural analogs of the SRS-A LTs and of LTB4, we have recently determined a number of the structural bases for the biological function of each moiety. With regard to the SRS-A leukotrienes, analogs differed from the native structures in the position of the peptide side chain and/or the hydroxyl group, the number and position of ethylenic bonds, the chirality at optically-active centers, or the structures of the four polar substituents in the C-1 to C-6 region. Analogs of LTB4, differing in the stereochemistry of their ethylenic bonds, were evaluated for chemotactic activity both in vitro, using human neutrophils, and in vivo intracutaneously in the rhesus monkey. We propose that true receptors exist on the pulmonary parenchyma of the guinea pig for the SRS-A LTs and on the primate neutrophil for LTB4. Further, LTC4, LTD4 and LTE4 have been shown to elicit a wheal and prolonged flare in human skin, whereas LTB4 evokes a time-dependent induration. The interaction of these secondary mediators may be critical to a fully developed host inflammatory response to both immunologic and non-immunologic injury. PMID- 6284664 TI - Messenger function of prostaglandins in cell to cell interactions and control of proteinase activity in the rheumatoid joint. AB - Destruction of joint structures in arthritis may result from failure of normal mechanisms controlling interactions among cells of the various tissues of the joint. Normal synovium in culture produces less prostaglandin E (PGE) and collagenase than rheumatoid. When rheumatoid synovium is dissociated into cells, the adherent cell cultures rapidly lose the ability to synthesize large amounts of PGE and collagenase and become indistinguishable from normal synovial cells. A mononuclear cell factor (MCF) derived from supernatant media of cultured human blood mononuclear cells and a 'synovial factor(s)' (SF) from cultures of either normal or rheumatoid synovial fragments both stimulate production of PGE and proteinase by cells derived from human synovium, cartilage and bone. The activities of factors which may be present in these stimulatory supernatants may be unmasked in vitro when they are removed from the normal control present in vivo. Normal synovium probably contains cells which, with the appropriate stimulus, may be recruited to participate in joint tissue degradation. Normal connective tissue turnover may also be controlled by a neutral metallo-proteinase inhibitor (TIMP), which is produced in considerable amounts by normal synovium, but which cannot be detected in cultures of rheumatoid synovium. While corticosteroids inhibit the production and action of MCF and SF, they stimulate production of TIMP by normal or rheumatoid synovial tissue in vitro and may contribute to the endogenous control mechanisms. PGE may also have a modulatory role in these cellular interactions. PMID- 6284665 TI - Disease states in genetic complement deficiencies. PMID- 6284666 TI - Multiple cutaneous granular cell tumors. PMID- 6284667 TI - Some applications of salivary gland scintigraphy. AB - Some of the uses of technetium 99 pertechnetate in salivary gland investigation are shown. By means of a gamma camera, which records dynamic studies, normal salivary physiology is investigated and also various forms of salivary gland pathology. The use of salivary scintigraphy in evaluation of the end results of parotid duct repositioning is discussed. This does not appear to have been reported previously. PMID- 6284668 TI - Value of sialography and scintigraphy in diagnosis of salivary gland disorders. AB - The purpose of this study was to examine the diagnostic value of sialographic and scintigraphic tests conducted to determine changes in salivary gland function and structure in patients with salivary gland disorders. A simple sialographic technique using a catheter fitted over a lacrimal probe was performed. For optimal pantomographic study of the salivary glands, a slightly oblique view with the head turned to the side was demonstrated to be of value. Examination of the salivary glands by sialography reveals the detailed morphology of the duct system. Sialograms were evaluated for filling and secretory phases and major ducts were also studied morphologically. Clearing radiographs indicated that glands with impaired function have prolonged clearing times. The contrast sialogram is most useful in evaluating recurrent inflammatory disorders, and the radiosialogram finds its greatest value in evaluating patients with suspected neoplasms. These examinations should not be used in lieu of a histologic diagnosis in tumor cases, except when a smooth margined radiopositive lesion corresponding with a clinically evident Warthin's tumor is noted. Perhaps the most important feature of salivary scintigraphy is its sensitivity in depicting changes in glandular dysfunction. In this study objective documentation provided by the scintigraphy closely parallels the results of sialography. PMID- 6284669 TI - Augmentation of the edentulous mandible using bone and hydroxyapatite: a comparative study in dogs. AB - This study compared autogenous bone, freeze-dried homografts, and hydroxyapatite in interpositional grafting of the mandible in beagle dogs. Healing was assessed radiologically and histologically. Results showed that the rate of incorporation of the autografts was greater than that of the homografts, and at both 4 and 12 weeks healing was more advanced. There was also rapid healing around the hydroxyapatite grafts, which became firmly bonded to the surrounding bone. The apatite was not resorbed, and new trabeculae of bone had been deposited onto much of its surface. This investigation confirms that hydroxyapatite is well tolerated and may be a suitable substitute for autogenous bone for augmentation of the atrophic mandible. PMID- 6284670 TI - Acinic cell carcinoma of the sublingual gland. Scintigraphy in pre-operative evaluation. PMID- 6284671 TI - Radical intermediates involved in the bleaching of the carotenoid crocin. Hydroxyl radicals, superoxide anions and hydrated electrons. AB - The participation of the primary radicals in the bleaching of aqueous solutions of the carotenoid crocin by ionizing radiation was investigated, employing both X radiolysis and pulse radiolysis. The pulse-radiolytic data demonstrated a very rapid diffusion-controlled attack by both hydroxyl radicals (.OH) and hydrated electrons (eaq-), while superoxide anions (O2-) did not react at all. The site of the initial reaction of these radicals was not limited to the polyene chromophore. Slower secondary reactions involving crocin alkyl or peroxy radicals contribute mainly to the overall bleaching, in particular during steady-state irradiation. PMID- 6284672 TI - Effect of gamma-radiation on functioning of bean hypocotyl mitochondria: lipids and lipid-dependent enzymes of the electron transfer chain (ETC). PMID- 6284673 TI - The effect of apple fibre on diabetic control and plasma lipids. PMID- 6284674 TI - Cefotaxime clinical trial results in septicaemic patients in Ireland and the U.K. PMID- 6284675 TI - Microbiological aspects of cefotaxime. PMID- 6284677 TI - Clinical experience with cefotaxime with particular reference to septicaemia. PMID- 6284676 TI - Cefotaxime in the management of acute lower respiratory tract infection. PMID- 6284679 TI - [Heck's focal epithelial hyperplasia]. AB - Three patients with focal epithelial hyperplasia Heck are presented. In two cases biopsies were made and investigated biochemically and by electron microscopy. In one case typical papillomavirus and HPV 1-DNA could be demonstrated, in the other the attempts of isolation failed. Virologic results and epidemiologic considerations suggest a viral etiology of focal epithelial hyperplasia Heck, but they are not sufficient to prove this. PMID- 6284678 TI - Captopril, an orally active angiotensin I converting enzyme inhibitor in the treatment of renovascular and essential hypertension. AB - The hypotensive response to captopril is described for 12 hypertensive patients, 7 of whom had renovascular hypertension. The drug was effective in lowering blood pressure. The few reversible adverse reactions that occurred included loss of the sense of taste in one patient and rash and fever in another. Three patients with renal failure showed deterioration of renal function during treatment, suggesting the advisability of treating such cases with lower dosages. PMID- 6284680 TI - Reconstruction of the columella with nasolabial flaps. AB - A one-stage operation is described for reconstruction of large defects of the columella nasi, the lower part of the nasal septum, and the vestibular floors. Nasolabial flaps are transported on a cranial, de-epithelialized pedicle, and passed deep to the nostrils into the vestibular cavity. The method is illustrated with four case reports. PMID- 6284681 TI - The role of nuclear scanning in head and neck surgery. AB - A number of procedures in nuclear medicine are available to assist the surgeon in evaluating pathologic conditions of the head and neck. Gallium-67 citrate scanning is occasionally useful in determining the extent of head and neck tumors but rarely can it detect an occult tumor and it cannot distinguish between an infection and a tumor. Technetium-99m pertechnetate can be used to evaluate diseases of the salivary glands including 123 and technetium-99m pertechnetate are commonly used to obtain images of the thyroid, but some malignant thyroid nodules will accumulate technetium-99m pertechnetate and the suspicious character of the nodule may be only appreciated by iodine scanning. The use of iodine 131 is generally limited to the detection of a substernal thyroid, therapeutic thyroid ablation, and the treatment of hyperthyroidism and thyroid cancer. Bone scanning has been used experimentally to evaluate the healing of mandibular bone grafts and to detect local extension of primary head and neck tumors to skull and facial bones. PMID- 6284682 TI - Use of triton WR-1339 in cytochemical and biochemical characterization of phytolysosomes. AB - A new cytochemical assay of acid phosphatase activity employing Triton WR-1339, a non-ionic detergent, has been used to demonstrate the probable origin of the phytolysosome system and of the organization of the Golgi complex in the root meristem, Cucurbita pepo. The results were controlled biochemically by differential and linear density gradient ultracentrifugation of extracts from normal and Triton-treated roots. The various fractions were characterized by electron microscopy or by assay of marker enzymes. PMID- 6284683 TI - Variations in epidermal cytochrome oxidase activity after local irradiation. AB - Cytochrome oxidase activity was evaluated histochemically as an index of mitochondrial damage after local irradiation with X-rays. It was determined by microphotometry on the tail skin of newly born Wistar rats four days after irradiation with doses ranging from 2 to 16 krad. The enzyme activity of the whole epidermis increased after irradiation, the increases being related to the increase in thickness of the epithelium which was observed as a response to irradiation injury. Within the dose range tested, the enzyme concentration (expressed per uni volume of tissue) decreased in relation to the dose applied. At the electron microscopy level, the cytochemical demonstration of cytochrome oxidase revealed an irregular reaction over the cristae, intramitochondrial vacuolization and partial homogenization of the matrix. Positive membrane fragments were seen around lipid droplets. This reaction confirms the mitochondrial origin of these previously observed radiation-induced vacuoles. PMID- 6284684 TI - Ultrastructural localization of adenosine diphosphatase activity in cultured aortic endothelial cells. AB - Electron microscopy cytochemical studies demonstrate that the plasma membrane of cultured aortic endothelial cells contain significant amounts of adenosine diphosphatase. The activity is due to an ectoenzyme on the upper surface of the cell. Intracellular activity was noted in multilamellar bodies. PMID- 6284685 TI - Localization and properties of ATPase activity in pea stems and wheat coleoptiles. AB - Microsomal fractions from wheat coleoptiles and pea stems contain a microsomal ATPase activity that requires divalent cations (Ca2+ is more effective than Mg2+) and shows further stimulation by KCl. The effects of added indoleacetic acid were inconclusive. Cytochemical studies on both species showed most pronounced staining for ATPase in the plasmalemma at pH 7.0. However, at pH 5.5, the coleoptile cells showed heaviest staining for ATPase in the endoplasmic reticulum and dictyosomes. The results are discussed with regard to the postulated role of ATPase activity in relation to proton pumping and plant cell elongation. PMID- 6284686 TI - Effect of training on beta-adrenergic receptor number in rat heart. AB - Female Sprague-Dawley rats were subjected to endurance-training programs, and the effect of training on myocardial beta-adrenergic receptor number, receptor binding characteristics, and adenylate cyclase (AC) activities associated with the receptor were examined. Training produced a 45% (P less than 0.01) increase in the succinate dehydrogenase activity of the plantaris muscle. Specific (-) [3H]dihydroalprenolol (DHA)-binding data were subjected to Scatchard plot analysis to quantify beta-adrenergic receptor number and DHA-binding characteristics of myocardial membranes. The DHA concentrations at which 50% of the total binding sites were occupied were similar for membranes from sedentary (1.95 +/- 0.51) and trained (1.59 +/- 0.34 nM) groups. Total DHA-binding sites of membranes from control (91.6 +/- 13.3) and trained (83.1 +/- 7.6 fmol/mg) groups were also similar. Basal and maximally stimulated AC activities were also unchanged by endurance training. Fluoride-stimulated AC activities of crude homogenate and 10,000 g fractions decreased 47 and 49%, respectively, with training. No differences were observed in a 40,000 g fraction. The specific activities of a ouabain-sensitive Na+-K+-ATPase (a sarcolemmal membrane marker) of crude homogenate, 10,000 g, and 40,000 g membrane fractions were similar. These data indicate that training produces no detectable difference in the potential for adrenergic responses at the receptor level. PMID- 6284687 TI - Effect of aerobic conditioning on cardiovascular response to isometric exercise. AB - The response to isometric handgrip exercise (IHE) and isometric quadriceps exercise (IQE) (30% maximum voluntary contraction held 3 min) was studied before and after 5 wk of aerobic training. Training exercises involved only the lower extremities. Seventeen healthy unconditioned males aged 21--35 yr were subjects. During training nine subjects received propranolol in doses that provided a high degree of beta-adrenergic blockade; eight received a placebo. All subjects were tested before training or drug and after training, 3--5 days off drug. With IGE after training, the placebo group had lower maximum heart rate (91 +/- 4 to 79 +/ 5 beats/min, P less than 0.05), systolic blood pressure (151 +/- 5 to 139 +/- 4 mmHg, P less than 0.05), and double product (heart rate x systolic blood pressure) (138 +/- 10 x 10(2) to 110 +/- 7 x 10(2), P less than 0.05). The response to IQE in the propranolol group was unchanged after training. Heart rate, systolic blood pressure, and double product with IHE was unchanged in either group after training. Aerobic conditioning can modify the hemodynamic response to isometric exercise. This effect is specific for the trained muscle group and is prevented by beta-adrenergic blockade. PMID- 6284688 TI - Concurrent generalized amyloidosis and infectious peritonitis in a cat. PMID- 6284689 TI - Field evaluation of a canine parvovirus vaccination program, using feline origin modified live virus vaccine. AB - Antibody titers measured by hemagglutination inhibition testing were determined in previously vaccinated dogs at the time of booster vaccination and 2 weeks later. All vaccines consisted of modified live panleukopenia virus. The booster injection was administered approximately 6 months after the initial parvovirus vaccination series was given. Fecal and serum specimens were collected immediately before and 2 weeks after administration of the booster vaccine for hemagglutination and hemagglutination inhibition testing, respectively. All dogs were privately owned and were from the Columbus, Ohio, area but were from environments with various exposure potentials to canine parvovirus. Results of hemagglutination (HA) testing on feces were negative in all dogs before and after booster vaccination. Therefore, these vaccinations did not interfere with interpretation of HA testing of feces. Results of serum hemagglutination inhibition (HI) testing indicated that 50% of the dogs had serum titers less than 1:80 prior to vaccination and that, of these dogs, 65.2% still had serum titers less than 1:80 2 weeks after the booster vaccination. Only 10.9% of all dogs had a marked increase in serum HI titer after the booster vaccination, indicating that overall serologic response to vaccination was poor. High HI titers (greater than or equal to 1:640) were associated with exposure to other dogs and cats in the neighborhood or to dogs suspected of having had parvovirus infection. PMID- 6284690 TI - Percutaneous fixation of left displaced abomasum, using a bar suture. PMID- 6284691 TI - The cells of Boettcher in the bat, Pteronotus p. parnellii. AB - The structural characteristics, distribution and intercellular relationships of the cells of Boettcher were studied in the mustache bat, Pteronotus P. parnellii. The cells of Boettcher have many structural features similar to those described in other mammals, but in Pteronotus they are distributed throughout the cochlea and are associated with relatively large amounts of secretory and/or absorptive material. Much of this material seems to be derived from or contribute to, a darkly staining upper layer of the basilar membrane. This material accumulates in elaborate microvillus-filled intercellular channels which are restricted to an area near the basilar membrane. The channels communicate with the basilar membrane surface through wide intercellular spaces and through small canals. The microvillus-filled channels are confluent with large extracellular spaces between Boettcher's cells and a single row of cells which form the floor of the outer tunnel. The latter have irregular shaped nuclei, contain many vacuoles and like Boettcher's cells, are associated with large amounts of basilar membrane-like material. Observations on Pteronotus, as well as other species of bats, do not support concepts relating Boettcher's cells to hair cell innervation patterns or to high frequency hearing. PMID- 6284692 TI - The outlook for antifungal prophylaxis in the compromised host. PMID- 6284693 TI - Antirhinovirus drugs. PMID- 6284694 TI - Virulence of mecillinam-resistant spherical mutants of Klebsiella pneumoniae and Escherichia coli. PMID- 6284695 TI - Randomised, comparative trial of mecillinam, mecillinam/ampicillin and chloramphenicol in the treatment of enteric fever. PMID- 6284696 TI - The comparative activities of N-formimidoyl theinamycin (MK0787), moxalactam, cefotaxime and cefoperazone against Yersinia enterocolitica and Listeria monocytogenes. PMID- 6284697 TI - High pressure liquid chromatographic determination of aflatoxins in peanut butter using a silica gel-packed flowcell for fluorescence detection. AB - A high pressure liquid chromatographic method has been developed for determining aflatoxins B1, B2, G1, and G2 in peanut butter. The method is based on extraction with acidified aqueous methanol, partition of the aflatoxin into methylene chloride, and purification of the extract on a 2 g silica gel column. The extracted aflatoxins are resolved on a microparticulate (10 micrometer) porous silica gel column in ca 10 min with a water-washed chloroform-cyclohexane acetonitrile solvent that contains 2% isopropanol. The fluorescence detection system determines aflatoxins B1, B2, G1, and G2 at low levels, i.e., 0.25 ppb B1, 0.5 ppb G1, and 0.2 ppb B2 and G2. Multiple assays of 5 samples of naturally contaminated peanut butters containing total aflatoxins (B1 + B2 + G1 + G2) at levels of 1, 2, 3, 9, and 17 ppb gave intralaboratory coefficients of variation of 7, 4, 4, 11, and 3%, respectively. Samples spiked at levels of 5, 9, and 17 ppb total aflatoxins showed recoveries of 79, 81, and 81%, respectively. PMID- 6284698 TI - The effect of dietary fibre constituents (pectin and guar gum) on oral glucose tolerance. PMID- 6284699 TI - Surface enhancement of sporulation and manganese oxidation by a marine bacillus. AB - In a seawater medium containing 0.005% yeast extract and 0.005% peptone, a marine bacillus, SG-1, sporulated only when associated with solid surfaces. The spores (rather than the vegetative cells) were responsible for the oxidation of manganese, and the degree of sporulation was determined by the surface area available rather than by the chemical nature of the clay or silica surface used. PMID- 6284700 TI - Transposition of Tn7 occurs at a single site on the Caulobacter crescentus chromosome. AB - Insertion of Tn7 in nine independently derived isolates occurred in a single small region of the Caulobacter crescentus chromosome, suggesting that Tn7 insertion occurs at or near a single site. The order of markers in this region of the Caulobacter chromosome was shown to be flaA-argG-Tn7. PMID- 6284701 TI - Molecular cloning of four tricarboxylic acid cyclic genes of Escherichia coli. AB - A fragment of DNA (3.1 kilobases [kb]) from a ColE1 Escherichia coli DNA hybrid plasmid containing the bacterial citrate synthase gene (gltA) was subcloned in both orientations into phage lambda vectors by in vitro recombination. The resulting phages were able to transduce gltA and, as prophages, complemented the lesion of a gltA mutant, showing that a functional gltA gene is contained in the 3.1-kb fragment. The segment of E. coli DNA cloned in these lambda gltA phages was extended in vivo by prophage integration and aberrant excision in the gltA region. Plaque-forming derivatives, carrying up to three additional tricarboxylic acid cycle genes, succinate dehydrogenase (sdh), 2-oxoglutarate dehydrogenase (sucA), and dihydrolipoamide succinyltransferase (sucB), were isolated and characterized by their transducing and complementing activities with corresponding mutants, and the order of the genes was confirmed as gltA-sdh-sucA sucB. Physical maps of a variety of the transducing phages showed that the four tricarboxylic acid cycle genes are contained in a 12.8-kb segment of bacterial DNA. The four gene products, plus a possible succinate dehydrogenase small subunit, were identified in postinfection labeling studies, and the polarities of gene expression were defined as counterclockwise for gltA and clockwise for sdh, sucA, and sucB, relative to the E. coli linkage map. PMID- 6284702 TI - A chromosomally located transposon in Pseudomonas aeruginosa. AB - A new transposon, Tn2521, coding for carbenicillin, streptomycin, spectinomycin, and sulfanilamide resistance, has been identified in Pseudomonas aeruginosa. The transposon occurs naturally in the chromosome of clinical strains of P. aeruginosa isolated in geographically separated hospitals. This has been demonstrated by its transductional linkage to the pur-136 marker and also by Southern hybridization. Tn2521 is 6.8 kilobases, can transpose from the chromosome to both IncP-1 and IncP-2 plasmid genomes, and has a pattern of restriction endonuclease sites unlike that of any previously described transposon. The carbenicillin resistance carried by Tn2521 is due to the PSE-4 type of beta-lactamase. PMID- 6284703 TI - Isolation and properties of Bacillus brevis mutants unable to produce tyrocidine. AB - Mutants of Bacillus brevis ATCC 10068 were isolated which produced less than 1/100 of the amount of tyrocidine produced by the parent strain. These mutants produced spores at the same frequency and which were as resistant to heating at 80 degrees C for up to 3 h as were those produced by the parent strain. A partially purified tyrocidine synthetase from strain ATCC 10068 catalyzed [32P]PPi-ATP exchange reactions dependent on added tyrocidine-constituent amino acids. These activities were separated into three groups (I, II, and III) by fractionation on an Ultrogel AcA34 column. Each group was similar to one of the three components (heavy, intermediate, and light, respectively) found previously for strain ATCC 8185 except that glutamate-dependent activity was not detected in the group I activities and some amino acyl-tRNA synthetase activities were associated with the group III activities. Some of the mutants were shown to have defective tyrocidine synthetase enzymes. Mutant BH30 was defective in two of the group II amino acid-dependent [32P]PPi-ATP exchange reactions, mutant BH16 was defective in one of the group I and one of the group II reactions, and mutant BH34 had alterations to activities in all of the groups. It is unlikely that any of these mutants could synthesise tyrocidine. We conclude that tyrocidine is not involved in either the sporulation process or the resistance of spores of B. brevis ATCC 10068 to heating at 80 degrees C for up to 3 h. PMID- 6284704 TI - DHA system mediating aerobic and anaerobic dissimilation of glycerol in Klebsiella pneumoniae NCIB 418. AB - In Klebsiella pneumoniae NCIB 418, the pathways normally responsible for aerobic growth on glycerol and sn-glycerol 3-phosphate (the glp system) are superrepressed. However, aerobic growth on glycerol can take place by the intervention of the NAD-linked glycerol dehydrogenase and the ATP-dependent dihydroxyacetone kinase of the dha system normally inducible only anaerobically by glycerol or dihydroxyacetone. Conclusive evidence that the dha system is responsible for both aerobic and anaerobic dissimilation of glycerol was provided by a Tn5 insertion mutant lacking dihydroxyacetone kinase. An enzymatically coupled assay specific for this enzyme was devised. Spontaneous reactivation of the glp system was achieved by selection for aerobic growth on sn-glycerol 3 phosphate or on limiting glycerol as the sole carbon and energy source. However, the expression of this system became constitutive. Aerobic operation of the glp system highly represses synthesis of the dha system enzymes by catabolite repression. PMID- 6284705 TI - Mutations affecting regulation of methionine biosynthetic genes isolated by use of met-lac fusions. AB - Fusions of the lac genes to the promoters of four structural genes in the methionine biosynthetic pathway, metA, metB, metE, and metF, were obtained by the use of the Mu d(Ap lac) bacteriophage. The levels of beta-galactosidase in these strains could be derepressed by growth under methionine-limiting conditions. Furthermore, growth in the presence of vitamin B12 repressed the synthesis of beta-galactosidase in strains containing a fusion of lacZ to the metE promoter, phi(metE'-lacZ+). Mutations affecting the regulation of met-lac fusions were generated by the insertion of Tn5. Tn5 insertions were obtained at the known regulatory loci metJ and metK. Interestingly, a significant amount of methionine adenosyltransferase activity remained in the metK mutant despite the fact that the mutation was generated by an insertion. Several Tn5-induced regulatory mutations were isolated by screening for high-level beta-galactosidase expression in a phi(metE'-lacZ+) strain in the presence of vitamin B12. Tn5 insertions mapping at the btuB (B12 uptake), metH (B12 dependent tetrahydropteroylglutamate methyltransferase), and metF (5,10-methylenetetrahydrofolate reductase) loci were obtained. The isolation of the metH mutant was consistent with previous suggestions that the metH gene product is required for the repression of metE by vitamin B12. The metF::Tn5 insertion was of particular interest since it suggested that a functional metf gene product was also needed for repression of metE by vitamin B12. PMID- 6284706 TI - Cloning of antibiotic resistance and nutritional genes in streptomycetes. AB - Methodology which allows consistent shotgun cloning of streptomycete genes is presented. Parameters that increase transformation efficiency of Streptomyces lividans 66 were adjusted to generate reproducibly a population of cloned genes likely to represent the entire genome. Factors which influence the recovery of viable transformants include: growth phase of the mycelium, ionic and osmotic characteristics of the medium during protoplast formation and transformation, and moisture content and protoplast density during regeneration. A modified transformation procedure was devised which increased transformation frequency more than 20-fold (allowing up to 10(7) primary transformants per microgram of SLP1.2 covalently closed circular DNA) and greatly facilitated the cloning of drug resistance genes and biosynthetic genes, using one of two plasmid vectors. Viomycin resistance genes on BamHI or PstI fragments were cloned from S. vinaceus genomic DNA into S. lividans, using the SLP1.2 vector. At least three different S. vinaceus BamHI fragments (1.9, 5.8, or 8.5 kilobases) confer viomycin resistance; only one PstI fragment (4.3 kilobases) was found. Recombinant plasmids were all able to produce lethal zygosis and to be transferred by conjugation within S. lividans. SCP2 was used to clone S. coelicolor A3(2) genes that "complemented" the auxotrophic mutation hisD3, argA1, or guaA1. Recombinant DNA technology can now be applied to economically and academically interesting problems unique to streptomycete molecular biology. PMID- 6284707 TI - Biochemical characterization of resistance determinants cloned from antibiotic producing streptomycetes. AB - Determinants of antibiotic resistance have been cloned from four antibiotic producing streptomycetes into Streptomyces lividans. Biochemical analyses of resistant clones revealed the presence of enzymes that had previously been characterized as likely resistance determinants in the producing organisms. These included: 23S rRNA methylases from S. azureus and S. erythreus, which confer resistance to thiostrepton and erythromycin, respectively; viomycin phosphotransferase from S. vinaceus; and aminoglycoside phosphotransferase and acetyltransferase from the neomycin producer S. fradiae. In general, the levels of antibiotic resistance of the clones were similar to those of the producing organisms. Although the two aminoglycoside-modifying enzymes from S. fradiae could independently confer only low-level resistance to neomycin, the presence of both enzymes in the same strain resulted in a level of resistance comparable with that of the producing organism. PMID- 6284708 TI - Cloning and expression of the distal portion of the histidine operon of Escherichia coli K-12. AB - The operator-distal genes hisBHAFI(E) of the Escherichia coli K-12 histidine operon were mapped on a DNA fragment 4,500 base pairs long. This fragment, originally present in a lambda transducing phage, was cloned in the vector plasmid pBR313. A restriction map was determined, allowing identification of the orientation of the genes in the fragment. The cloned genes were expressed in appropriate hosts, independent of the orientation of the DNA fragment, as shown by transformation tests and by enzyme assays of one of the gene products, hisB, histidinol phosphatase. An internal transcription initiation site was identified by isolation of the cellular RNA, hybridization to specific DNA probes, and mapping by S1 nuclease. PMID- 6284709 TI - thiK and thiL loci of Escherichia coli. AB - Mutants of Escherichia coli K-12 auxotrophic for thiamine phosphates were produced in stepwise fashion from the polyauxotrophic F- strain JC1552, via intermediate production of thiamine auxotrophs that had lost the enzymatic activity of either phosphomethylpyrimidine kinase or thiamine phosphate pyrophosphorylase. They include two types: one responds to thiamine monophosphate or thiamine pyrophosphate, and the other responds to thiamine pyrophosphate only; the former lacks thiamine kinase activity, and the latter lacks thiamine monophosphate kinase activity, in addition to the enzymatic defects caused by the first mutations. We found two genes, for which we propose the designations thiK and thiL, which govern the activities of thiamine kinase and thiamine monophosphate kinase, respectively. By conjugation and P1 transduction, the thiK locus was mapped at about 25 min, between pyrC and purB and close to fabD. The relative order of thiK with respect to nearby genes was tentatively established as pyrC-ptsG-fabD-thiK-purB. In the case of thiL, the locus was situated at about 9 min, between tsx and acrA and probably 0.2 min clockwise from the former. PMID- 6284710 TI - A gene and its product required for transposition of resistance transposon Tn2603. AB - Tn2603 is a multiple-resistance transposon encoding resistance to ampicillin, streptomycin, sulfonamide, and mercury and having a molecular size of 20 kilobase pairs, with 200-base-pair inverted repeats at both ends. The essential sites and functions of Tn2603 which are required for its transposition were determined through construction and characterization of various deletion mutants affecting the efficiency of transposition. Deletions were introduced in plasmid pMK1::Tn2603 by partial digestion with restriction endonuclease EcoRI in vitro. Analysis of deletion mutants showed that the inverted repeat segments at both ends of the trans-acting diffusible product(s) encoded in the right-hand side of the central portion were required for the transposition of Tn2603. An essential gene product was revealed as a protein having a molecular weight of 110,000 by analysis of polypeptides synthesized in Escherichia coli minicells. This protein was assumed to be the so-called transposase. PMID- 6284711 TI - Role of proton motive force in genetic transformation of Bacillus subtilis. AB - This study explored the role of the proton motive force in the processes of DNA binding and DNA transport of genetic transformation of Bacillus subtilis 168 strain 8G-5 (trpC2). Transformation was severely inhibited by the ionophores valinomycin, nigericin, and 3,5-di-tert-4-hydroxybenzylidenemalononitrite (SF 6847) and by tetraphenylphosphonium. The ionophores valinomycin and nigericin also severely inhibited binding of transforming DNA to the cell envelope, whereas SF-6847 and carbonylcyanide-p-trifluoromethoxyphenylhydrazone hardly affected binding. The proton motive force, therefore, does not contribute to the process of DNA binding, and valinomycin and nigericin interact directly with the DNA binding sites at the cell envelope. The effects of ionophores, weak acids, and tetraphenylphosphonium on the components of the proton motive force and on the entry of transforming DNA after binding to the cell envelope was investigated. DNA entry, as measured by the amount of DNase I-resistant cell-associated [3H]DNA and by the formation of DNA breakdown products, was severely inhibited under conditions of a small proton motive force and also under conditions of a small delta pH and a high electrical potential. These results suggest that the proton motive force and especially the delta pH component functions as a driving force for DNA uptake in transformation. PMID- 6284712 TI - Involvement of cyclic AMP and its receptor protein in filamentation of an Escherichia coli fic mutant. AB - Cyclic AMP (cAMP) inhibited septum formation in Escherichia coli PA3092 and induced cell filamentation at elevated temperatures. This phenomenon was first observed in E. coli PA3092 and is due to a temperature-sensitive mutation. We tentatively named this mutation fic (filamentation induced by cAMP). The fic gene was located near rpsL (formerly strA) on the E. coli K-12 map. the inhibitory effect of cAMP on cell division and filamentation in a fic mutant was not observed in a crp mutant. When cAMP was removed from the culture medium, filaments were divided into rods as the intracellular cAMP level decreased. These results suggest that the cAMP-cAMP receptor protein complex causes filamentation in the fic mutant, E. coli PA3092. PMID- 6284713 TI - Nucleotide sequence analysis of the complement resistance gene from plasmid R100. AB - The multiple antibiotic resistance plasmid R100 renders Escherichia coli resistant to the bactericidal action of serum complement. We constructed a plasmid (pOW3) consisting of a 1,900-base-pair-long restriction fragment from R100 joined to a 2,900-base-pair-long fragment of pBR322 carrying ampicillin resistance. E. coli strains carrying pOW3 or R100 were up to 10,000-fold less sensitive to killing by serum complement than were plasmid-free bacteria or bacteria carrying pBR322. Nucleotide sequencing revealed that 875 of the 1,900 bases from R100 correspond exactly to part of the bacterial insertion sequence IS2. The remaining 1,075 bases contained only one sizeable open reading frame; it covered 729 base pairs (243 amino acids) and was preceded by nucleotide sequences characteristic of bacterial promoters and ribosome binding sites. The first 20 amino acids of the predicted protein showed features characteristic of a signal sequence. The remainder of the predicted protein showed an amino acid composition almost identical with that determined for the traT protein from the E. coli F factor. Southern blot analysis showed that the resistance gene from R100 does not hybridize to the serum resistance gene from ColV,I-K94 isolated by Binns et al.; we concluded that these genes are distinct. PMID- 6284714 TI - ColE1 copy number mutants. AB - A deletion mutant of the colicin E1-derived plasmid, pDMS6642, exhibited an approximately fourfold increase in copy number. We subsequently isolated hydroxylamine-induced mutants of that plasmid that had a further increase in copy number. Analysis of them suggests that the increased copy number of pDMS6642 is associated with transcriptional readthrough from a Tn3 transposon into the region of ColE1 containing information that influences plasmid replication. The hydroxylamine mutation in one copy number mutant appeared to increase the plasmid copy number by stimulating readthrough transcription from the Tn3 transposon into the ColE1 replication control region, whereas the other hydroxylamine mutation acts by another mechanism. PMID- 6284715 TI - Genetic mapping with Tn5-derived auxotrophs of Caulobacter crescentus. AB - Chromosomal insertions of Tn5 in Caulobacter crescentus displayed complete stability upon transduction and proved useful in strain building on complex media. RP4-primes constructed in vitro containing C. crescentus genomic sequences in the HindIII site of the kanamycin resistance gene failed to show enhanced or directed chromosome mobilization abilities. One of these kanamycin-sensitive RP4 derivatives, pVS1, was used as a mobilization vector in conjugation experiments on complex media where chromosomal Tn5 transfer to the recipient was selected. pVS1-mediated transfer of Tn5-induced auxotrophic mutations occurred at frequencies of 10(-6) to 10(-8) per donor cell. During conjugation with Tn5 encoded kanamycin resistance as the selected marker, Tn5 remained in its donor associated locus in 85 to 100% of the transconjugants. A collection of eight temperature-sensitive donor strains bearing Tn5 insertion mutations from various regions of the C. crescentus genetic map were used to provide a rapid means for the determination of the map location of a new mutation. Use of the techniques described in this paper allowed an expansion of the C. crescentus genetic map to include the relative locations of 32 genes. PMID- 6284716 TI - Genetic and enzymatic characterization of the inducible glycerol dissimilatory system of Neurospora crassa. AB - The glycerol dissimilatory system in Neurospora crassa was analyzed through the characterization of 18 Glp- mutants which were isolated after inositol-less death and filtration enrichment. All mutants obtained by this procedure could be assigned to one of three complementation groups. The subsequent genetic characterization of these glp mutations revealed lesions on the I, II, and VI chromosomes at the glp-1, glp-2, and glp-4 loci, each of which was subjected to fine-structure analysis. Evidence from the enzymatic characterization of these mutants indicated that glp-2 and glp-4 were the structural genes encoding the mitochondrial glycerol-3-phosphate dehydrogenase and cytosolic glycerokinase, respectively. Additional evidence, obtained from studies of the inducibility of glycerokinase by glycerol, cold treatment, or deoxyribose, suggests that glp-1 is involved in controlling the expression of glp-4. PMID- 6284718 TI - Transcription initiation at the tryptophanase promoter of Escherichia coli K-12. AB - Restriction fragments containing the region preceding the tryptophanase structural gene, tnaA, were used as templates for in vitro transcription experiments. A transcription initiation site was detected that was dependent on the catabolite gene activator protein (CAP) plus cyclic AMP (cAMP). The mRNA produced in vitro was fingerprinted, and the nucleotide at which transcription was initiated was localized to the vicinity of two guanine residues 316 and 318 base pairs upstream of tnaA. A region exhibiting extensive difold symmetry and homology to the CAP binding site adjacent to the lactose operon promoter exists approximately 60 base pairs preceding the site of transcription initiation. Two HinfI restriction sites are located in this region. Restriction enzyme cleavage at these sites was prevented when DNA containing the promoter region was preincubated with CAP and cAMP. RNA polymerase was incapable of protecting these sites against this cleavage. CAP and cAMP addition did not protect against cleavage at a DdeI restriction site located in the -20 region of the promoter. RNA polymerase did protect against DdeI cleavage but only in the presence of CAP and cAMP. Thus, transcription initiation at the tryptophanase promoter involves cAMP-dependent, CAP-facilitated binding of RNA polymerase to the DNA. PMID- 6284719 TI - Characteristic lipids of Bordetella pertussis: simple fatty acid composition, hydroxy fatty acids, and an ornithine-containing lipid. AB - The lipids and fatty acids of Bordetella pertussis (phases I to IV) were analyzed by thin-layer chromatography, gas-liquid chromatography, and mass spectrometry and compared with those of B. parapertussis and B. bronchiseptica. The major lipid components of the three species were phosphatidylethanolamine, cardiolipin, phosphatidylglycerol, lysophosphatidylethanolamine, and an ornithine-containing lipid. The ornithine-containing lipid was characteristic of the genus Bordetella. The fatty acid composition of the total extractable cellular lipids of B. pertussis was mostly hexadecanoic and hexadecenoic acids (90%) in a ratio of about 1:1. The hexadecenoic acid of B. pertussis was in the cis-9 form. The fatty acid composition of the residual bound lipids was distinctly different from that of the extractable lipids, and residual bound lipids being mainly 3 hydroxytetradecanoic, tetradecanoic, and 3-hydroxydecanoic acids, with 3 hydroxydodecanoic acid occurring in some strains. It was determined that the 3 hydroxy fatty acids were derived from lipid A. The fatty acid composition of the total extractable cellular lipids of B. parapertussis and B. bronchiseptica, mainly composed of hexadecanoic and heptadecacyclopropanoic acid, differed from that of B. pertussis. Although the fatty acid composition of the residual bound lipids of B. parapertussis was similar to that of the residual bound lipids of B. pertussis, 2-hydroxydodecanoic acid was detected only in the bound lipids of B. bronchiseptica. PMID- 6284717 TI - Isolation of the kanamycin resistance region (Tn2350) of plasmid R1drd-19 as an autonomous replicon. AB - We have isolated a circular form of Tn2350, an IS1-flanked kanamycin resistance transposon forming part of the plasmid R1drd-19. This circle (pTn2350::9.6 kilobases) contains a single IS1 element and probably arises by recombination between the two directly repeated Is1 sequences of Tn2350. It can be used to transform Escherichia coli to kanamycin resistance. It is capable of autonomous replication but is not maintained stably in dividing cells and segregates under nonselective conditions. Cloning of a segment of pTn2350 on a conditional plasmid vector allowed us to assign the replication functions of this plasmid to a 1.6 kilobase restriction fragment. The plasmid R1drd-19 can thus be considered as a cointegrate between two replicons separated by IS1 sequences. PMID- 6284721 TI - The use of tricyclic antidepressants in chronic gastrointestinal pain. PMID- 6284720 TI - Doxepin effects on chronic pain, depression and plasma opioids. AB - Thirty patients with chronic low back or cervical pain combined with clinical depression were studied in a six-week, randomized, double-blind comparison of doxepin and placebo. Dependent variables included Hamilton Depression Scores, the Clinical Global Assessment Scale, and Profile of Mood States (POMS), and subjective ratings (visual analogue scales) of pain severity, percent of time pain felt, and effect of pain on activity, muscle tension, sleep, mood, and analgesic drug consumption. Plasma levels of doxepin, desmethyldoxepin, beta endorphin, and enkephalin-like activity were also measured. Significant improvements in the doxepin-treated group compared to the placebo group were seen in Hamilton scores, Global Assessment Scale, Profile of Mood States, percent of time pain felt, and effect of pain on sleep, muscle tension, and mood. Some improvement was observed after 1 week, although most improvement occurred at 6 weeks, when the mean doxepin dose was 2.5 mg/kg and plasma doxepin and desmethyldoxepin averaged 70 ng/ml. Nonspecific enkephalin-like activity (but not beta-endorphins) increased for the treatment group and decreased for the placebo group. The efficacy of doxepin compared with that of placebo was thus documented in several depressive and pain parameters, indicating that doxepin is a valuable treatment for patients with chronic pain and depression. PMID- 6284722 TI - Characterization of electron donation to cytochrome c-555 in Chromatium vinosum from ferrocyanide, tetramethylphenylenediamine and reduced dimethylquinone. Effects of redox potential, pH and salt concentration. AB - 1. The dependences of the reduction of ferricytochrome c-555 in the reaction center-cytochrome c complex on the redox potential and pH were investigated using N,N,N',N'-tetramethyl-p-phenylenediamine (TMPD), ferrocyanide, and reduced 2,5 dimethyl-p-quinone as electron donors. 2. In the reduction of cytochrome c-555 by TMPD, the unprotonated form was the exclusive electron donor to the cytochrome with a second-order rate constant of 1.0 X 10(5) M-1.s-1. 3. Ferrocyanide reduced cytochrome c-555 slowly with a rate constant of 7.8 X 10(3) M-1.s-1 at infinite salt concentration. The value of -5.2 X 10(-4) elementary charge/A2 was estimated as the surface charge density in the vicinity of cytochrome c-555 by analyzing the salt effect on the cytochrome reduction using the Gouy-Chapman theory. 4. The characteristics of the dependences of the reduction of cytochrome c-555 by reduced 2,5-dimethyl-p-quinone on the redox potential and pH were well explained by the redox potential and pH dependences of the formation of the semiquinone. In the neutral-to-alkaline pH range the anionic semiquinone was the main electron donating species with a second-order rate constant of 6.0 X 10(7) m-1.s-1. PMID- 6284723 TI - Effects of pH on membrane fluidity of human erythrocytes. AB - The effects of pH on the membrane fluidity of intact human erythrocytes, ghosts, and their lipid vesicles were studied by spin label techniques in the range of pH 3.0 to 9.1. Two fatty acid spin labels, 5-nitroxide stearic acid (5NS) and 12 nitroxide stearic acid (12NS), and a maleimide spin label were used for the labeling of the membrane lipids and proteins, respectively. The outer hyperfine splitting (T parallel) was measured as a parameter of membrane fluidity. In the case of 5NS, the T parallel values for intact erythrocytes and ghosts remained almost constant over the entire pH range at 22 degrees C but those for their lipid vesicles changed slightly, indicating the vertical displacement of the labels in lipid bilayers. On the other hand, the ESR spectra of 12NS incorporated into intact erythrocytes and ghosts, as compared with their lipid vesicles, showed marked pH dependence. By means of spin labeling of membrane proteins, the conformational changes of the proteins were observed in the pH range mentioned above. These results suggest a possible association between the strong pH dependence of the T parallel values and the conformation changes of membrane proteins. The pH dependence of the membrane fluidity was also investigated in cholesterol-enriched and -depleted erythrocytes. The effects of cholesterol demonstrated that the membrane fluidity was significantly mediated by cholesterol at low pH, but not at high pH. PMID- 6284724 TI - The interaction between calmodulin and microtubule proteins. IV. Quantitative analysis of the binding between calmodulin and tubulin dimer. AB - We reported previously that calmodulin binds to tubulin in a Ca2+-dependent manner, thereby inhibiting microtubule assembly. In this work, we quantitatively investigated the binding between calmodulin and tubulin by applying two analytical methods. One was the frontal analysis using affinity chromatography originally developed by Kasai and Ishii (J. Biochem. 84, 1061-1069, 1978). The use of tubulin-Sepharose columns gave a dissociation constant of 4.0 microM. The other was the equilibrium gel filtration developed by Hummel and Dreyer (Biochim. Biophys. Acta 63, 532-534, 1962). This method using a Sephadex G-100 column provided a dissociation constant of 3.5 microM under the same medium conditions as in the frontal analysis, and it was found that 2 mol calmodulin could bind to 1 mol tubulin. Furthermore, the frontal analysis method was convenient for studies on the effect of temperature and ionic strength on the binding. Upon elevating the temperature, the dissociation constant increased. Increase in the ionic strength also increased the dissociation constant. PMID- 6284725 TI - Isolation and partial characterization of two distinct DNA topoisomerases from cauliflower inflorescence. AB - DNA topoisomerases which remove superhelical turns in closed circular DNA have been isolated from cauliflower inflorescences using polyethylene glycol fractionation, ammonium sulfate precipitation, and column chromatography on CM Sephadex or CM-cellulose and DNA-cellulose. Two distinct enzymes, topoisomerase-I and ATP-dependent topoisomerase, were separated clearly by CM-Sephadex or CM cellulose, and partially characterized using agarose gel electrophoresis with plasmid pBR322 DNA. Topoisomerase-I acts like other eucaryotic DNA topoisomerases in the absence of ATP, is stimulated by spermidine and inhibited by EDTA. The ATP dependent topoisomerase acts like topoisomerase-I only in the presence of ATP in the reaction medium, is inhibited by spermidine and EDTA, and does not introduce supertwists into closed duplex DNA or produce catenate aggregates under the present reaction conditions. PMID- 6284726 TI - Affinities of various nucleases to DNA-Sepharose under non-digestive conditions: survey for productive affinity chromatography. AB - 1. It has been reported that DNase I can be highly purified from pancreas extract by affinity chromatography on a dDNA-Sepharose column under non-digestive conditions. In the present study, the adsorption-elution of other nucleases on the column under non-digestive conditions was studied. 2. All the seven kinds of nucleases tested were adsorbed when applied on a dDNA-Sepharose column under conditions which did not allow the enzymes to hydrolyze the DNA. The non digestive conditions were as follows. i) For DNase II (pI=10.2), pH 3.0 in the presence of 50 mM sodium sulfate (inhibitor), ii) for micrococcal nuclease (pI=9.6), pH 4.0 in the absence of Ca2+ (activator), iii) for restriction endonucleases Eco RI (pI=5+1), Hind III (pI=5+1), and Bam HI (pI=5+1), pH 4.0 in the presence of 20% glycerol and 0.1% Neopeptone (stabilizers), and iv) for nucleases S1 (pI=5+1) and nuclease P1 (pI=4.5), pH 7.0. At the respective pH's, the enzymes other than nucleases S1 and P1 were cationic so as to exhibit electrostatic attraction to the anionic dDNA-Sepharose. Although S1 and P1 were anionic, they still adsorbed to the column. 3. All the adsorbed nucleases described above were eluted by a concentration gradient of KCl without changing pH. The ionic strengths required for elution were 0.19 for DNase II, 0.53 for micrococcal nuclease, 0.73 for Eco RI, 0.72 for Hind III, 0.37 for Bam HI, 0.17 for P1, and 0.13 for S1. The fact that the ionic strength required for the elution of DNase I (pI=5.0) was 0.39 at pH 4.0 indicates that the former five enzymes except DNase II can be chromatographed with almost the same or higher efficiency than DNase I, because the proteins adsorbed with no-specific affinity could be mostly eluted at lower ionic strength. On the other hand, the fact that nucleases P1 and S1 were adsorbed in spite of electrostatic repulsion suggests that these two enzymes can also be effectively chromatographed, especially when other cationic proteins are previously removed by an appropriate method such as adsorption to a typical cation exchanger. PMID- 6284727 TI - Purification of cardiac sarcolemmal vesicles: high sodium pump content and ATP dependent, calmodulin-activated calcium uptake. AB - Highly purified vesicles of cardiac sarcolemma were prepared from a homogenate of canine ventricular muscle by density gradient centrifugation. The preparation showed an extremely high content of (Na+,K+)-ATPase. The steady state levels of Na+-dependent phosphoenzyme formation in the presence of Triton X-100 and the specific ouabain binding in the absence of Triton X-100 were, respectively, 773 and 907 pmol.mg-1 under the optimum conditions. On the other hand, the amount of Ca2+-dependent phosphoenzyme formed in the absence of Triton X--100 was less than 2 pmol.mg-1. This demonstrates that the preparation was virtually free of contaminant sarcoplasmic reticulum fragments. The preparation showed ATP dependent Ca2+ uptake. Almost all the Ca2+ accumulated on the addition of ATP was rapidly released by the subsequent addition of NaCl. This finding gives evidence that the ATP-driven Ca2+ pump exists in the cardiac sarcolemma. The Ca2+ uptake was unaffected by 2 microM digitoxin, 1 microM monesin, and 200 microM dinitrophenol. These results exclude the possibility that transmembrane gradients of Na+ and H+ were involved in this Ca2+ uptake. The Ca2+ pump was activated by calmodulin. The concentration of calmodulin giving a half-maximum activation was 0.05 micrograms.ml-1, which is equivalent to 3 nM. This activation was removed by addition of trifluoperazine, a specific inhibitor of calmodulin. PMID- 6284728 TI - Restriction map of double-stranded DNA copy synthesized from poliovirus Sabin 1 RNA. AB - Nearly full-sized double-stranded cDNA was prepared from virion RNA of poliovirus Sabin 1 (LSc, 2ab) strain using reverse transcriptase. The double-stranded cDNa was cleaved at four and two sites by the restriction endonucleases Bam HI and Hind III, respectively. Based on the cleavage patterns of double-stranded cDNA into segments of various lengths, each of which had the sequence corresponding to that of the 3'-end of the viral genome, the location of each restriction fragment was determined. The cDNA fragments were cloned with pBR322 as a vector and some of their nucleotide sequences were determined. The DNA sequence indicated that the restriction map obtained was consistent with the known arrangement of viral RNA fragments (1-3). Comparison of the nucleotide sequences of the cloned Hind III (460 bases) and Pst I (434 bases) fragments with those of the corresponding region of Mahoney type 1 genome (3) revealed 12 point-mutation sites. PMID- 6284729 TI - High dimensional structure of the antigen-binding site of anti-viomycin immunoglobulin analyzed by enzyme immunoassay. AB - Precise immunological recognition of anti-viomycin antiserum at detailed parts in the structure of viomycin was studied by cross reactivities of the antiserum to viomycin and its ten analogs using an enzyme immunoassay of viomycin. The antiserum clearly recognized all minor modifications in the sixteen membered ring of viomycin, indicating that the antiserum clearly recognizes the whole structure of the sixteen membered ring. Recognition of the antiserum on the beta-lysine terminus was also examined showing that the antiserum was also recognized on this part. Thus, the anti-viomycin antiserum was deduced to recognize the whole structure of viomycin, from which the deduction was made that the anti-viomycin antibodies in the antiserum must possess cavities fitting the whole structure of viomycin. The crystal dimensions of viomycin are 13 A in length, 8 A in width, and 7 A in depth. Thus, the high dimensional structure of the binding sites of the anti-viomycin antibodies was deduced to possess cavities of a similar size to that of viomycin. PMID- 6284730 TI - Effect of phosphorylation of porcine cardiac troponin I by 3':5'-cyclic AMP dependent protein kinase on the actomyosin ATPase activity. AB - 1. Porcine cardiac native tropomyosin was phosphorylated by bovine cardiac 3':5' cyclic AMP-dependent protein kinase. Most of the phosphate incorporation was observed in troponin I, the maximum of which was 0.7 mol of Pi per mol of troponin I. 2. In the presence of phosphorylated native tropomyosin, actomyosin ATPase activity was 15-40% lower than that in the presence of the unphosphorylated preparation at all calcium ion concentrations (1.5 x 10(-8) M 2.4 x 10(-5) M). Half-maximum activation of ATPase was obtained with a concentration of 7 x 10(-7) M Ca2+ (unphosphorylated) and 1.3 x 10(-6) M Ca2+ (phosphorylated), respectively. Maximum ATPase activity was reached with 3 x 10( 6) M Ca2+ (unphosphorylated) and 1.0 x 10(-5) M Ca2+ (phosphorylated). 3. Porcine cardiac troponin I isolated by affinity chromatography inhibited ATPase activity of desensitized actomyosin in the presence of tropomyosin. There was little difference between phosphorylated troponin I and a control preparation with regard to the inhibitory effect of ATPase activity. 4. Troponin C from rabbit skeletal muscle neutralized the inhibitory effect of troponin I. The minimum amount of troponin C required for complete neutralization was approximately equimolar to troponin I. The inhibitory effect of phosphorylated troponin I was neutralized by troponin C less effectively than that of unphosphorylated preparation. PMID- 6284731 TI - Incorporation of spin-labeled ganglioside analogues into cell and liposomal membranes. AB - Ganglioside analogues (gangliosides) with an electron spin resonance label in a long aliphatic hydrocarbon chain were used to investigate the possible insertion of the sialoglycolipid into the plasma membrane of cells. Three types of ESR signals observed in the labeled glycolipids were distinguished. They characteristically indicate an isotropic tumbling motion of spin label in solution, the micellar state of the glycolipid, and an anisotropic motion in a lipid bilayer. Below CMC, gangliosidoide carrying one aliphatic hydrocarbon chain showed an isotropic tumbling motion. After the gangliosidoide had been incubated with liposomes or blood cells, there was an immediate change to an ESR signal showing an anisotropic motion. The signal was typical of the spin-label in liposomes prepared in the presence of spin-labeled sialoglycolipid. It can be concluded that the gangliosidoide was inserted into the lipid phase of liposomal or cellular membranes from the incubation medium. The overall splitting (2A parallel) of 5SL-gangliosidoides in membranes was larger than those of 5SL galactosylceramide, 5SL-phosphatidylcholine, and 5SL-stearic acid, though the 2A parallel of 12SL-gangliosidoide was almost the same as those of other lipids having a nitroxide group in the 12-position of an acyl chain. This indicates that the head group movement is restricted in gangliosidoide molecules. PMID- 6284732 TI - Modifying effect of vinblastine on superoxide anion production by membrane perturbing agents in polymorphonuclear leukocytes. AB - Superoxide anion production in peritoneal polymorphonuclear leukocytes obtained from guinea pigs was stimulated by in vitro treatment with membrane-perturbing agents, such as cytochalasin D, concanavalin A, phorbol myristate acetate, myristate, digitonin, and NaF. Vinblastine modified these stimulating effects on the superoxide anion production, but its modifying effect was not uniform. The effect of cytochalasin D was stimulated by vinblastine at the concentration of 10(-5)-10(-7) M, whereas it was inhibited at the concentration of 10(-4) M. At 10(-4)-10(-5) M, vinblastine was inhibitory to the effect of concanavalin A, and lower concentrations had no significant effect. Stimulation of the superoxide anion production by phorbol myristate acetate and myristate was further enhanced by vinblastine at any concentration in the range of 10(-4)-10(-8) M with peaks at 10(-6) and 10(-5) M, respectively. Vinblastine had little effect on the stimulation of the superoxide anion production by digitonin and NaF throughout the concentration range examined. The mechanism of the interaction of these membrane-perturbing stimulants and vinblastine is discussed. PMID- 6284733 TI - Mitochondrial chemiluminescence elicited by acetaldehyde. AB - Acetaldehyde-dependent chemiluminescence has been found to be a sensitive technique for the study of superoxide and hydrogen peroxide formation in beef heart mitochondria. The system responds to ATP and antimycin A with increased emission intensities and to ADP and rotenone with decreased intensities, indicating that the chemiluminescence reflects the energy status of the mitochondrion. These effects are based on the ability of acetaldehyde to react with superoxide and hydrogen peroxide to form metastable intermediates which decay spontaneously with the emission of light. Additionally, these intermediates can react with cyanide to give alternative products which can also decay with the emission of light, the cyanide-evokable chemiluminescence. The interaction of acetaldehyde with mitochondria is complex because acetaldehyde can serve as a hydrogen source for NADH and as an inhibitor (at high concentration) of electron transport, and appears to be a reducing agent for a heat-stable site that autoxidatively generates HOOH from O2-.. Inasmuch as acetaldehyde is a metabolite of ethanol, this broad spectrum of reactivity may play a role in the hepatic and cardiac toxicity that is associated with alcoholism. The heat-stable site that generates HOOH from O2-. has been studied further and appears to contain vicinal dithiol which is primarily responsible for the cyanide-evokable chemiluminescence. PMID- 6284734 TI - Effect of pyridine homologues on proton flux through the CF0 . CF1 complex and photophosphorylation in chloroplasts. AB - At concentrations below 1 mM, hydrophobic pyridine homologues decrease the rate of photophosphorylation and light-stimulated hydrolysis of ATP and light activated exchange of the tightly bound nucleotides in chloroplasts, but increase the rate of the Hill reaction. Unlike uncoupling agents, the presence of the organic base at such low concentrations decreases the rate of light-dependent leakage and has no effect on the efficiency of two-stage photophosphorylation in broken chloroplasts. By assuming that the organic base is bound to independent equivalent sites in the thylakoid membrane, a simple expression can be derived which relates the observed rates of photophosphorylation and light-stimulated hydrolysis of ATP quantitatively to the concentration of the organic base in solution and gives dissociation equilibrium constants which are on the order of the relative hydrophobicities of the pyridine homologues. A possible mechanistic model for the CF0 . CF1 complex is proposed which could serve as the basis for a unified interpretation of the kinetics of proton translocation in illuminated chloroplasts, the steady-state rate of photophosphorylation, the light-stimulated ATPase activity, and the light-activated exchange of tightly bound adenine nucleotides. PMID- 6284735 TI - Dephosphorylation of synthetic phosphopeptides by protein phosphatase-T, a phosphothreonyl protein phosphatase. AB - The synthetic phosphohexapeptides Arg-Arg-Ala-Thr(35P)-Val-Ala and Arg-Arg-Ala Ser(32P)-Val-Ala, phosphorylated by the cAMP-dependent protein kinase and differing only in the nature of the phosphorylated residue, have been used as substrates of a partially purified rat liver protein phosphatase-T, distinct from the multifunctional protein phosphatase-1. While the phosphothreonyl hexapeptide is readily dephosphorylated (exhibiting a Km = 15 microM), the phosphoseryl one is almost unaffected. Such a behavior is not shared by protein phosphatase-1, calf intestine alkaline phosphatase, and potato acid phosphatase, all of which are more active on the phosphoseryl hexapeptide. The NH2-terminal basic residues critical for cAMP-dependent phosphorylation are not required in the dephosphorylation reaction, as both Arg can be removed without impairing the efficiency of protein phosphatase-T toward the phosphothreonyl peptide. On the other hand, the replacement of 2 Pro for the Ala and Val flanking Thr(32P), to give a new phosphohexapeptide reproducing the phosphorylated site of protein phosphatase inhibitor-1, prevents the protein phosphatase-T activity. Moreover, IgG heavy chain 32P labeled in tyrosine is not affected by protein phosphatase-T, while it is dephosphorylated by alkaline phosphatase. These results would indicate that protein phosphatase(s)-T represent a distinct class of protein phosphatases specifically involved in the dephosphorylation of phosphothreonyl residues fulfilling definite structural requirements. PMID- 6284736 TI - The site of attachment of retinal in bacteriorhodopsin. The epsilon-amino group in Lys-41 is not required for proton translocation. AB - Chymotryptic fragments C-1 (amino acids 72-248) and C-2 (amino acids 1-71) of bacteriorhodopsin have been shown previously to reassociate so as to regenerate the native bacteriorhodopsin chromophore in lipid/detergent mixtures and to form functional proton-translocating vesicles. The fragment C-2 has now been selectively methylated with formaldehyde and sodium cyanoborohydride to give the epsilon-dimethylamino derivatives of Lys-30, 40, and 41 in 96-99% average yield. The methylated and unmethylated C-2 fragments were identical in their ability to reassociate with fragment C-1 and retinal to regenerate the bacteriorhodopsin chromophore and to form functional proton-translocating vesicles. In contrast, dimethylation of the lysine residues of the C-1 fragment gave a derivative which did not form an active complex with unmethylated C-2. We conclude that the epsilon-amino group in Lys-41 is not required for Schiff's base formation with retinal at any step in the light-driven proton-translocation cycle. PMID- 6284737 TI - Specificity of the effect of lipoxygenase metabolites of arachidonic acid on calcium homeostasis in neutrophils. Correlation with functional activity. AB - The ability of the major neutrophil-derived lipoxygenase metabolites of arachidonic acid to increase the rate of 45Ca influx in rabbit neutrophils was examined. The results obtained demonstrate that (5S),(12R)-dihydroxy-6,8,11,14 (cis,trans,trans,cis)-eicosatetraenoic acid (leukotriene B4) is the most active of the arachidonic acid metabolites. The activity of leukotriene B4 is highly stereospecific in that its three nonenzymatically derived isomers are essentially inactive. The omega-hydroxylation of leukotriene B4 results in a compound that is nearly as active as leukotriene B4 as far as its ability to stimulate calcium influx and neutrophil aggregation while being a much weaker secretagogue. The further conversion of leukotriene B4 into a dicarboxylic acid removes all detectable biological activity. 5,6-Oxido-7,9,11,14-eicosatetraenoic acid (leukotriene A4) methyl ester was also found to increase the rate of calcium influx, while the degradation products of native leukotriene A4 were essentially inactive. These results demonstrate that a close correlation exists between the ability of the various lipoxygenase products to alter calcium homeostasis in rabbit neutrophils and their biological activities. PMID- 6284738 TI - Inactivation of glucocorticoid-binding capacity by protein phosphatases in the presence of molybdate and complete reactivation of dithiothreitol. AB - Glucocorticoid receptor in rat liver cytosol is inactivated (rendered unable to bind steroid) by incubation with calf intestine alkaline phosphatase or highly purified rabbit muscle phosphoprotein phosphatase (phosphorylase phosphate, protein phosphatase C). The receptor is inactivated by both enzymes even when 10 mM sodium molybdate is present. Receptors that are inactivated by phosphatases in the presence of molybdate can be reactivated to the steroid-binding state by addition of dithiothreitol, but receptors that are inactivated in the absence of molybdate cannot be reactivated. These observations suggest that dephosphorylation leads to oxidation of a moiety (-SH) on the receptor that is required for steroid binding. Molybdate apparently preserves the receptor in a form such that reduction returns the receptor to the steroid binding state. We would propose that molybdate may act by complexing with sulfur groups on the receptor. PMID- 6284739 TI - Siderophore iron transport followed by electron paramagnetic resonance spectroscopy. AB - Siderophore iron transport was followed in Ustilago sphaerogena using isotope transport assays coupled with EPR spectroscopy. EPR spectroscopy was used as a quantitative tool to follow the rate of reduction of siderophore iron(III) to iron(II) in the cell suspension by following the disappearance of the signal at g = 4.3. This rate was compared with the rate of iron transport, measured by the disappearance of radioactively labeled iron from the medium. The transport of three iron chelates was examined: the ferric siderophores ferrichrome and ferichrome A, and iron(III) chelated to excess citrate. For the transport of ferrichrome, an iron(III) ionophore, the rate of reduction of iron(III) to iron(II) was significantly lower than the rate of uptake of isotope from the medium supernatant, which is consistent with the established mechanism of uptake of the entire complex followed by intracellular reduction to remove the iron from the ligand. However, the rate of reduction of ferrichrome A, a non-ionophore, was identical with the rate of transport of iron into the cell. Iron(III) citrate was reduced at a rate slightly lower than the rate of transport. These data suggest that reduction of iron(III) is involved in the transport of iron from ferichrome A and possibly from iron(III) citrate. PMID- 6284740 TI - Nuclear precursor molecules of the two beta-globin mRNAs in Friend erythroleukemia cells. AB - Processing of the beta major and beta minor globin pre-mRNAs has been compared in murine erythroleukemia cells induced to synthesize hemoglobin by dimethyl sulfoxide or hemin treatment, using both the Northern blot technique and S1 nuclease mapping with 3' and 5' end-labeled probes. The small intervening sequence of both beta-globin pre-mRNAs was removed in one step, although minor amounts of incompletely spliced RNA were detected. During the processing of the large intervening sequence of beta major globin pre-mRNA two internal splice sites were clearly detected. On the contrary, the beta minor globin pre-mRNA did not show any internal splice sites. A model of processing of the mouse adult beta major globin pre-mRNA is proposed. PMID- 6284741 TI - Altered G-protein glycosylation in vesicular stomatitis virus-infected glucose deprived baby hamster kidney cells. AB - The glycosylation of the G-protein was analyzed in vesicular stomatitis virus infected baby hamster kidney cells incubated in the absence of glucose. The results indicate that the G-protein in glucose-starved cells is initially glycosylated from a lipid donor with a glucosylated oligosaccharide which is resistant to endo-beta-N-acetylglucosaminidase H and partially susceptible to alpha-mannosidase. With longer times, the protein-bound carbohydrate chain becomes much more sensitive to alpha-mannosidase while remaining endo-beta-N acetylglucosaminidase H-resistant. Purified virions from glucose-starved baby hamster kidney cells, labeled with [35S]methionine and isolated on a sucrose gradient, contain altered forms of the G-protein, whereas the other viral proteins remain unchanged. These altered forms could also be radiolabeled with [3H]mannose, and upon analysis of labeled glycopeptides by chromatography on concanavalin A-Sepharose and Bio-Gel P-6, it was apparent that modification of the oligosaccharide portion of the G-protein occurs in baby hamster kidney cells, leading to aberrant mature carbohydrate chains. PMID- 6284742 TI - Human ribonucleases. Quantitation of pancreatic-like enzymes in serum, urine, and organ preparations. AB - Antibodies against pure human pancreatic ribonuclease (RNase) were used to study ribonuclease levels in human tissues and body fluids. The antibodies completely inhibit the activity of purified RNase as well as ribonuclease activity in crude pancreatic extracts. RNase activity is inhibited by 70-80% in serum and urine, indicating that a significant proportion of the RNases in these preparations are structurally like the pancreatic enzyme. In contrast, inhibition of RNase activities from spleen (8%) and liver (30%) was inefficient suggesting that most of the RNases in these tissues are structurally unlike the pancreatic enzyme. A competitive binding radioimmunoassay (RIA), sensitive in the range of 1-100 ng of RNase, was developed to quantitate the pancreatic like enzymes. The RIA of crude tissue preparations and samples fractionated by gel filtration was compatible with inhibition results. Enzymes structurally like pancreatic RNase could be quantitated despite the presence of other RNase activities. Immunological quantitation of pancreatic like RNases was also found to be much more simple and precise than enzymatic assays comparing RNA and polycytidylate substrates. We suggest the immunological assays will be useful in the quantitation and definition of tissue of origin of RNases in serum of patients with pancreatic carcinoma. PMID- 6284743 TI - Extraction of proteins from the large subunit of bovine mitochondrial ribosomes under nondenaturing conditions. AB - The 55 S mammalian mitochondrial ribosome (referred to hereafter as "mitoribosome") is protein-rich, containing nearly twice as much protein as the Escherichia coli ribosome. In order to produce soluble mitochondrial proteins and protein-deficient subribosomal particles for use in functional and structural studies, the proteins of bovine mitoribosomes were extracted by washing in a series of buffers containing increasing concentrations of LiCl as the only chaotropic agent. LiCl disruption is used in order to preserve the solubilized proteins in a substantially "native" configuration. The extraction mixtures were characterized by sucrose density gradient analysis and the compositions of the stripped protein and residual pellet fractions were determined by two-dimensional polyacrylamide gel electrophoresis. In order to analyze the behavior or individual proteins, the intensity of Coomassie blue stain for each protein was normalized against the intensity of stain for the same protein in a control sample. Buffers with 1, 2, and 4 M LiCl each extract a specific subset of mitoribosomal proteins, while another group of proteins remains in the residual pellet fraction. Although very few proteins are detected in only one condition, most proteins are specifically enriched in one fraction. This LiCl procedure, therefore, produces fractionated groups of mitoribosomal proteins which can be used directly as a source for those proteins in which they are enriched, or they can be used as a starting point in further purification procedures. In contrast to results with E. coli ribosomes, several mitoribosomal proteins remain core associated, indicating a different structural organization in these ribosomes. PMID- 6284744 TI - Subunit structure of Escherichia coli exonuclease VII. AB - Exonuclease VII has been purified 7,500-fold to 87% homogeneity from Escherichia coli K12 using a new purification procedure. The enzyme has been shown to be composed of two nonidentical subunits of 10,500 and 54,000 daltons. This has been confirmed by restoration of exonuclease VII activity after renaturation of denatured and purified subunits. The structure of the native enzyme consists of one large subunit and four small subunits. We have previously isolated exonuclease VII mutant strains containing defects which map at two distinct loci. Subunit-mixing experiments utilizing wild type enzyme and temperature-sensitive enzyme produced by an xseB mutant strain have shown that the xseB gene codes for the small subunit of the enzymes. PMID- 6284745 TI - Altered transport properties in Escherichia coli mutants selected for pH conditional growth on 3-deoxy-2-oxo-D-gluconate. PMID- 6284746 TI - Hormonal regulation of fructose-6-P-2-kinase and fructose-2,6-P2 by two mechanisms. PMID- 6284748 TI - A phospholipid requirement for dolichol pyrophosphate N-acetylglucosamine synthesis in phospholipase A2-treated rat lung microsomes. AB - The role of phospholipids in the activity of UDP-Glc-NAc:dolichol phosphate GlcNAc-1-phosphate transferase of rat lung microsomes has been investigated. Treatment of microsomes with phospholipase A2 in the presence of delipidated bovine serum albumin resulted in a time-dependent loss of 65 to 75% of the enzyme activity and approximately 30% of the phospholipids. Addition of phosphatidylglycerol to the enzyme assay system containing phospholipase A2 treated microsomes restored activity to that obtained with native microsomes and phosphatidylglycerol. Addition of phosphatidylinositol, phosphatidylcholine, or cardiolipin resulted in only partial restoration of activity, whereas phosphatidylserine and phosphatidylethanolamine were without effect. Triton X-100 was not by itself capable of restoring activity, but was required for the phospholipid effect. Measurements of the phospholipase A2 hydrolysis products released from the microsomes during digestion, and other control experiments of adding fatty acids and lysophospholipids to the enzyme assay system, indicated that the loss of UDP-GlcNAc:dolichol phosphate GlcNAc-1-phosphate transferase activity was not due to product inhibition. PMID- 6284749 TI - Low density lipoprotein receptor binding in aging human diploid fibroblasts in culture. AB - High affinity cell surface receptors for low density lipoproteins (LDL) are inducible in cultured human lung fibroblasts by the removal of lipoproteins from the cell culture medium. The binding, uptake, and degradation of 125I-LDL by fibroblasts decrease with increasing number of population doublings. The affinity of LDL receptor binding, however, remained unchanged at different population doublings levels. Hence, the difference in LDL binding activity in the aging fibroblasts can be attributed to a reduction in the number of receptor sites on the cell membrane. Cellular uptake of [4-14C]cholesterol and 2-deoxy-D-[1 14C]glucose mediated through mechanisms independent of the LDL receptor pathway revealed no significant difference in early and late passage fibroblasts. This suggests that the alteration in the LDL receptor binding in serially passaged fibroblasts is an "age-related" phenomenon. The late population doubling fibroblasts require more LDL in the culture medium for feedback inhibition of LDL receptor synthesis. Thus, aging fibroblasts are both progressively less inducible and less suppressible in the regulation of their cell membrane LDL receptors. Similar results were also obtained with respect to the regulation of DL-3-hydroxy 3-methyl-glutaryl coenzyme A reductase in the aging fibroblasts in culture; the enzyme has become less inducible and less supressible as the fibroblasts approach the limit of their in vitro lifespan. These age-related alterations in the cellular metabolism of LDL and cholesterol might contribute to our understanding of the increased risk of athlerosclerosis in our aging population. PMID- 6284747 TI - The presence of a reducible disulfide bond in milk xanthine oxidase. AB - The stoichiometry of reducing equivalents per protomer for the complex molybdoflavoprotein xanthine oxidase has been re-examined by reductive titrations with sodium dithionite and anaerobic reoxidation with cytochrome c and phenazine methosulfate of dithionite- or photo-reduced enzyme. It is found that 8.0 +/- 0.1 reducing equivalents are taken up (or given up) by the enzyme, a value of 2 eq greater than expected on the basis of the known oxidation-reduction centers in the enzyme. The reaction of reduced xanthine oxidase with [14C]iodoacetate indicates that, in the reduced form of the enzyme, additional cysteine residues are available for reaction. These results, in conjunction with the observation that reaction of oxidized enzyme with sulfite results in the appearance of an additional equivalent of thiol capable of reacting with 5,5'-dithiobis-(2 nitrobenzoic acid) or iodoacetate, indicate the presence of a disulfide linkage in the enzyme that can be reduced by dithionite or photochemically employing EDTA and 5-deazaflavin. Neither xanthine nor lumazine, however, is capable of reducing this oxidation-reduction center, suggesting that the disulfide does not play a role in the catalytic reactions of the enzyme. These results resolve discrepancies in the literature which indicated that greater than 6 reducing equivalents were consistently needed to bring about the complete reduction of xanthine oxidase. PMID- 6284750 TI - The stereochemical course of nucleotidyl transfer catalyzed by ATP sulfurylase. AB - ATP sulfurylase catalyzes the synthesis of ATP from adenosine 5'-phosphosulfate and magnesium pyrophosphate with inversion of configuration at phosphorus. This implies an "in line" displacement mechanism in the ternary complex and effectively eliminates both an adjacent mechanism followed by pseudorotation and a double displacement mechanism involving an adenylyl-enzyme intermediate. The double displacement mechanism had been invoked previously to account for a number of observations, including the ability of the enzyme to catalyze the hydrolysis of MgATP to AMP and MgPPi, and the exchange of Mg32PPi into MgATP in the absence of sulfate. PMID- 6284751 TI - Bacteriophage T4 gene 45. Sequences of the structural gene and its protein product. AB - Bacteriophage T4 gene 45 codes for a protein whose functions are required for both T4 DNA replication and T4 late gene transcription. To facilitate studies of the interactions of 45 protein with the T4 DNA replication complex and with RNA polymerase, we have determined the primary structure of 45 protein. The amino acid sequence of 45 protein has been determined by correlating nucleotide sequence analysis of gene 45 with protein chemistry studies of 45 protein. Our studies indicate that gene 45 codes for a polypeptide containing 227 amino acids, with a calculated Mr = 24,710. The coding region of gene 46 is preceded by a putative promoter containing sequences which are homologous to Escherichia coli RNA polymerase recognition and binding regions. In addition, there are sequence similarities in the translation initiation regions of gene 45 and the rIIB gene, which may relate to their common regulation by regA protein. PMID- 6284752 TI - A requirement of bicarbonate for Ca2+-induced elevations of cyclic AMP in guinea pig spermatozoa. AB - Ca2+ causes less than 2-fold elevations of guinea pig sperm cyclic AMP concentrations when cells are incubated in a minimal culture medium in the absence of bicarbonate (HCO3-). However, in the presence of HCO3-, Ca2+ increases cyclic AMP by as much as 25-fold within 1 min. The (Ca2+, HCO3-)-induced elevations occur in either the presence or absence of the permeant anions, pyruvate and lactate. In the absence of extracellular Ca2+, HCO3- elevates cyclic AMP only slightly. The effect of HCO3- is concentration-dependent, with maximal responses obtained at concentrations of greater than 25 mM. Ca2+ (25 mM HCO3-) at concentrations of less than 100 microM causes one-half-maximal elevations of cyclic AMP. The (Ca2+, HCO3-)-induced elevations of cyclic AMP are observed at various extracellular pH values (7.5-8.5) and in the presence or absence of extracellular Na+ or K+. NH4Cl does not elevate sperm cyclic AMP concentrations and does not greatly alter the (Ca2+, HCO3-)-induced elevations. the putative Ca2+ transport antagonist, D-600 (100 microM), completely blocks the (Ca2+, HCO3 )-induced elevations of cyclic AMP. A23187, in the presence but not in the absence of extracellular Ca2+, increases sperm cyclic AMP but does not further elevate cyclic AMP in HCO3(-)-treated cells. These studies establish that Ca2+ dependent elevations of cyclic AMp in guinea pig spermatozoa are dependent on the presence of HCO3- and suggest that HCO3- is required for the uptake (exchange) or membrane sequestration of small amounts of physiologically active Ca2+. PMID- 6284753 TI - Secondary bioenergetic hypoxia. Inhibition of sulfation and glucuronidation reactions in isolated hepatocytes at low O2 concentration. AB - The O2 dependence of cytochrome oxidase, cellular ATP/ADP ratio, and drug sulfation and glucuronidation were studied in isolated hepatocytes to examine secondary biochemical changes of hypoxia related to decreased cytochrome oxidase function. A decrease in cytochrome oxidase function as well as in cellular ATP/ADP ratio occurred at low O2 concentrations and their O2 dependencies gave p50 values of 3.2 and 2.8 microM O2, respectively. Changes in the ATP/ADP ratio corresponded directly with changes in the oxidation-reduction state of cytochrome oxidase. The p50 value for sulfation of acetaminophen (2.5 microM O2) corresponded with the values for ATP/ADP ratio and cytochrome oxidase. Addition of compounds that lowered cellular ATP/ADP ratio caused a corresponding inhibition of sulfation. Selective use of ethionine, an adenosyl-trapping agent, demonstrated that the O2 dependence of sulfation related directly to a decrease in the absolute ATP concentration. The p50 value for glucuronidation (5.2 microM O2) was higher than that for sulfation, but in spite of this difference, glucuronidation also correlated well with cellular ATP/ADP ratio when this ratio was decreased by either O2 limitation or metabolic inhibitors. These results demonstrate that the predominant limitation of drug sulfation and glucuronidation during hypoxia is due to decreased cytochrome oxidase function. PMID- 6284754 TI - Contribution of oligosaccharide sulfation to the charge heterogeneity of a viral glycoprotein. AB - The single glycoprotein species of vesicular stomatitis virus exhibits heterogeneity in isoelectric focusing. Short term radioisotopic labeling of infected cells revealed that this heterogeneity is acquired after synthesis and glycosylation of the protein, during the period of oligosaccharide processing from the high mannose to the complex form. In this process, radioactive sulfate is incorporated randomly into different glycoprotein molecules, conferring unequal amounts of negative charge upon them. Since the virus glycoprotein is processed by cellular pathways, it is likely that differential sulfation is responsible for much of the charge heterogeneity displayed by cellular glycoproteins. PMID- 6284755 TI - Incorporation of amino acid analogs interferes with the processing of the asparagine-linked oligosaccharide of the MOPC-46B kappa light chain. AB - The incorporation of the leucine analog, beta-hydroxyleucine, or the isoleucine analog, 4-thiaisoleucine, into the MPOC-46B kappa chain interfered with the processing of the oligosaccharide moiety of newly synthesized light chains. Analog-containing chains were secreted with endo-beta-N-acetylglucosaminidase H sensitive oligosaccharides instead of the normal complex, endo-beta-N acetylglucosaminidase H-resistant oligosaccharide. The incorporation of the analogs did not significantly decrease the overall secretion of the MPOC-46B related proteins. These data form the basis for the hypothesis that the presence of the analog in the protein alters the peptide configuration sufficiently to prevent the processing of precursor high mannose oligosaccharides to complex oligosaccharides. PMID- 6284756 TI - Limited autolysis reduces the Ca2+ requirement of a smooth muscle Ca2+-activated protease. AB - Chicken gizzard smooth muscle contains large amounts of Ca2+-activated protease activity. Approximately 15 mg of purified enzyme can be obtained from 1 kg of fresh muscle. The enzyme consists of two subunits (Mr = 80,000 and 30,000) present in a 1:1 molar ratio. In the presence of CaCl2, the 80,000/30,000-dalton heterodimer (form I) is rapidly converted by limited autolysis to a 76,000/18,000 dalton species (form II). Both the 80,000- and 30,000-dalton subunits are degraded simultaneously. Moreover, the Ca2+ dependence for autolysis (K0.5 = 300 microM) is identical for both subunits. Neither the time course nor the Ca2+ dependence of the autolytic conversion reaction is altered by 10- and 20-fold molar excesses of substrate. Limited autolysis markedly reduces the Ca2+ requirement for substrate degradation. Using N-[ethyl-2-3H]maleimide-labeled 27,000-dalton cardiac myosin light chains as substrate, the Ca2+ requirement of form I was found to be quite high (K0.5 = 150 microM). Under similar conditions, the Ca2+ requirement of form II was 30-fold lower (K0.5 = 5 microM). Limited autolysis did not alter the specific activity of the enzyme. Our results demonstrate that smooth muscle contains an abundant amount of Ca2+-activated protease. Moreover, autolysis of this enzyme may play an important regulatory role by converting the native form to a species that is fully active at physiological levels of intracellular calcium ion. PMID- 6284757 TI - Increase of a surface glycoprotein by cyclic AMP in Chinese hamster ovary cells. Dependence on cell-cell interaction. AB - Cell surface proteins and glycoproteins of the CHO-K1 clone of Chinese hamster ovary cells were radiolabeled by surface-specific reactions as well as by metabolic incorporation of glycoprotein precursors. After the proteins were separated by two-dimensional gel electrophoresis, they were quantified according to the amount of radioactivity which was associated with individual proteins spots. A glycoprotein of 133,000 daltons (P133-11) wa found to increase severalfold by the elevation of the cytoplasmic level of adenosine 3':5'-cyclic monophosphate. In this paper, we report that the increase of P133-11 was more efficient in dense cultures than in sparse cultures. The use of conditioned medium or frequent medium changes during the treatment with the cyclic nucleotide did not influence the efficiency of the increase of the glycoprotein, indicating that the enrichment or exhaustion of diffusible materials was not responsible for this density-dependent phenomenon. The same conclusion was drawn from an experiment in which the amount of P133-11 was compared between homogeneously plated sparse cultures and cultures containing the same number of cells in the form of colonies with well contacted cells at an average density of 400 cells/colony. The increase of the protein was higher in the latter type of cell cultures. We, therefore, propose that the interaction of surface components upon cell-cell contact plays a role in the expression of P133-11 surface glycoprotein. PMID- 6284759 TI - Collaborative study of the assay for virus content of Marek's disease vaccines using a reference preparation. PMID- 6284758 TI - Hydrolases of pulmonary lysosomes and lamellar bodies. PMID- 6284760 TI - Interactions between saponin and 146S particles of foot-and-mouth disease virus. PMID- 6284761 TI - Qualitative assessment of 146 S particles of foot-and-mouth disease virus in preparations destined for vaccines. PMID- 6284762 TI - Release of 3H-adrenaline from an isolated intact preparation of the rabbit adrenal gland: no evidence for release modulatory alpha-adrenoreceptors. AB - 1 The possibility that catecholamine secretion from the rabbit adrenal gland is subject to modulation by a mechanism involving alpha-adrenoreceptors was investigated in an isolated preparation of the gland. 2 Intact left adrenal glands from the rabbit were perfused with Krebs-Henseleit solution through their vasculature. The adrenal catecholamine stores were radiolabelled with 3H adrenaline and subsequently, efflux of the radiolabel was elicited by stimulation of an attached segment of splanchnic nerve (60 s at 5 Hz). In some experiments, efflux of radiolabel was elicited by perfusion with potassium-enriched Krebs Henseleit solution (30 mM). 3 Radioactivity released in response to nerve stimulation was accounted for almost entirely by (3H)-adrenaline. Stimulation induced (S-I) efflux was abolished by 0.1 microM tetrodotoxin and by omission of calcium from the perfusion medium and was reduced by approximately 55% by 100 microM hexamethonium. 4 S-I efflux was enhanced to approximately 150% of control S-I efflux in the presence of 10 microM phenoxybenzamine; however, 3 microM phentolamine and yohimbine in concentrations of 0.1 and 1.0 microM had no effect; in a higher concentration (10 microM) yohimbine reduced S-I efflux by approximately 50%. It is likely that the effect of phenoxybenzamine in enhancing S-I efflux is due to blockade of reuptake of 3H-adrenaline since cocaine (30 microM) enhanced release to a similar extent. 5 S-I efflux was not altered in the presence of the alpha-adrenoreceptor agonists noradrenaline (0.1 microM), clonidine (1 microM) or oxymetazoline (10 microM). 6 Release of radiolabel evoked by perfusion with 30 mM potassium solution was unaltered in the presence of phentolamine (3 microM) of clonidine (1 microM). 7 These findings do not support the existence in the rabbit adrenal gland of a mechanism involving alpha adrenoreceptors through which catecholamine secretion may be modulated. PMID- 6284763 TI - The effects of intravenous 6-hydroxydopamine on peripheral alpha-adrenoreceptors. AB - 1 Peripheral alpha-adrenoreceptor-mediated responses were studied in conscious intact rabbits and in animals pretreated with intravenous 6-hydroxydopamine (50 mg/kg). 2 Treatment caused a large decrease in noradrenaline in plasma and peripheral tissue but only small falls in blood pressure. 3. There was a shift to the left in pressor dose-response curves to the alpha 1-adrenoreceptor agonist phenylephrine and the mixed alpha 1/alpha 2-adrenoreceptor agonist noradrenaline in 6-hydroxydopamine-pretreated animals. The alpha 1-adrenoreceptor antagonist prazosin was less effective at blocking the phenylephrine pressor response in these animals. 4 There was no change in pressor responses to the alpha 2 adrenoreceptor selective agonists clonidine and guanabenz and alpha adrenoreceptor antagonists were equally effective at blocking this pressor response in 6-hydroxydopamine-pretreated and in intact rabbits. 5 In conscious animals 6-hydroxydopamine pretreatment causes alteration in responses mediated by postsynaptic alpha 1-but not postsynaptic alpha 2-adrenoreceptors. These changes may help maintain blood pressure in the 6-hydroxydopamine treated rabbits. PMID- 6284764 TI - Adrenoreceptors in uterus. PMID- 6284765 TI - Experimental scoliosis in primates: a neurological cause. AB - Although a variety of techniques have been used with varying success to induce scoliosis in animals, primates have rarely been used. A series of monkeys is presented where scoliosis developed incidentally during the routine virulence testing of live, attenuated, oral poliomyelitis vaccines by intraspinal injection. The site and extent of histological damage in the different anatomical areas of the spinal cord were examined in 25 scoliotic monkeys and 25 matched controls. Analysis of the data demonstrated that there was significantly greater damage on the convex side of the spinal cords of the scoliotic animals, particularly in the sensory areas-the posterior horn and Clarke's column. Scoliosis was not thought to be caused by clinical poliomyelitis as the involvement of the anterior horn was not significantly greater than in the scoliotic animals than in the controls. These observations are taken to support the view that scoliosis may develop as a result of asymmetrical weakness of the paraspinal muscles due to the loss of proprioceptive innervation. PMID- 6284766 TI - 3,4-bis(3'-hydroxyphenyl)hexane--a new mammary tumor-inhibiting compound. AB - The syntheses of the hexestrol derivatives 3,4-bis-(3'-hydroxyphenyl)hexane (4a), 3,4-bis(4'-fluoro-3'-hydroxyphenyl)hexane (4b), 3,4 bis(3',4'dihydroxyphenyl)hexane (4c), and 3,4-bis(3',4'-diacetoxyphenyl)hexane (4d) are described. All compounds showed a marked, competitive inhibition of the estradiol receptor interaction (Ka4c greater than Ka4a greater than Ka4d greater than Ka4b). Evaluated in the mouse uterine weight test compounds 4c and 4d almost reached the estrone effect, whereas 4a and 4b did not produce full uterotrophic response. Compounds 4a--d antagonized the estrone stimulated uterine growth of the immature mouse. Compound 4a (NSC-297170) exhibited a specific, dose-related growth inhibition of the estrogen responsive MCF-7 human breast tumor cell line. Tested on the 9,10-dimethyl-1,2-benzanthracene-induced hormone-dependent mammary adenocarcinoma of the Sprague-Dawley rat all compounds showed marked inhibition of tumor growth. As in all experiments compounds 4a and 4b, which is resistant to hydroxylation in 4'-position exhibited an identical pattern of action, which is different from that shown by compound 4c, the effect of compound 4a cannot be explained by its possible catechol metabolite 4c. PMID- 6284768 TI - Simian virus 40 A gene function: further characterization and growth of tsA transformed chinese hamster cells. AB - Chinese hamster embryo cells transformed with the tsA 58 mutant of Simian virus 40 express the transformed phenotype at the permissive temperature (33 degrees C or 37 degrees C) and a "normal" phenotype at the nonpermissive temperature (40.5 degrees C). Immunofluorescence and immunoprecipitation of T antigens demonstrated that the "T" antigen (100 K) has an increase rate of synthesis and degradation at 40.5 degrees C. However, the cells continue to replicate at the nonpermissive temperature when assayed by flow cytometry and autoradiography. This DNA synthesis was cellular, not viral, and not owing to an increase in DNA repair. When the cell cycle distributions of G1, S, and G2 + M were assayed by the fraction labeled mitoses method, no differences were evident at the permissive and nonpermissive temperature; however, the doubling time was lengthened at 40.5 degrees C (13 hours vs. 100 hours). These results suggest that at 40.5 degrees C, the tsA transformed cells are cycling and dying. However, if the transformed cells are seeded onto monolayers of normal Chinese hamster cells at 40.5 degrees C, the cells are growth arrested when measured by growth assays, flow cytometry, autoradiography, and immunofluorescence for T antigen. Therefore, growth arrest can be obtained in tsA 58 transformed Chinese hamster cells when cocultured with normal Chinese hamster cells. PMID- 6284767 TI - Analysis of DNA attached to the chromosome scaffold. AB - Two different methods have been described to investigate whether any specific DNA sequences are intimately associated with the metaphase chromosome scaffold. The chromosome scaffold, prepared by dehistonization of chromosomes with 2 M NaCl, is a nonhistone protein complex to which many looped DNA molecules are attached (Laemmli et al., 1977, Cold Spring Harbor Symp. Quant. Biol. 42:351--360). Chromosome scaffold DNA was prepared from dehistonized chicken MSB chromosomes by restriction endonuclease EcoRI digestion followed by removal of the looped DNA by sucrose gradient sedimentation. Alternatively, the scaffold DNA was prepared from micrococcal nuclease-digested intact chromosomes using sucrose gradients containing 2M NaCl. Solution hybridization of the radioactively labeled scaffold DNA with a large excess of total nuclear DNA revealed that, in either case, the scaffold DNA is not a unique sequence class of genomic DNA. Southern-blotting hybridization also showed that the scaffold DNA prepared from EcoRI-digested dehistonized chromosomes was not enriched (or depleted) in the ovalbumin gene sequences. The possibility of a dynamic interaction of protein and DNA in the chromosome scaffold and the possibility that the scaffold is a preparative artifact are discussed. PMID- 6284769 TI - Autoradiographic detection and characterization of a Chinese hamster ovary cell mutant deficient in fucoproteins. AB - Autoradiography of colony replicas immobilized on filter paper was used to isolate a Chinese hamster ovary cell line deficient in incorporation of radiolabeled fucose into a trichloroacetic acid-insoluble fraction. This cell line, called 62.1, has the same growth rate at 37 degrees C as wild-type cells, but incorporates five times less fucose into acid-insoluble radioactivity. Chemical analysis of fucose bound to macromolecules also showed a fivefold reduction in the mutant. The fucoproteins of the mutant cell line differ qualitatively from those of wild-type cells as visualized by SDS gel electrophoresis fluorography; no differences were detected between total proteins as visualized by coomassie blue staining. The macromolecular sialic acid content of the mutant was somewhat higher than the wild type (20%). Studies of the synthesis of the glycoprotein of vesicular stomatitis virus in mutant and wild type cells showed that the mutant is unable to synthesize complex-type N-linked oligosaccharides. Enzyme assays show that ths defect in the mutant is due to reduction in UDP-N-acetylglucosamine-glycoprotein N-acetyl glucosaminyltransferase, a key enzyme in the assembly of complex glycopeptides. Hybridization studies have shown that mutant 62.1 has common mutations belonging to the same complementation group as mutant PhaR1-1. This latter mutant was previously isolated using lectin resistance by Stanley et al. (1975) and was also deficient in the above N-acetyl-glucosaminyltransferase. PMID- 6284770 TI - The increase in hormone-stimulated adenylate cyclase activity following Rous sarcoma virus transformation. AB - Rous sarcoma virus (RSV)-infected chicken embryo cells were used to study the effect of viral transformation on the hormone-stimulated synthesis of cyclic AMP. Transformation by RSV greatly increased the cells' ability to synthesize and accumulate cyclic AMP in response to the beta-adrenergic agonist isoproterenol as compared to untransformed cells. This enhancement was observed in both intact cells and in membranes prepared from these cells. The inclusion of guanosine 5'-0 (3-thiotriphosphate), a nonhydrolyzable analogue of GTP, in assays of adenylate cyclase activity did not abolish the quantitative differences between the transformed and normal cell membranes. Infection of cells by Rous-associated virus, which lacks the oncogene src, did not induce this hyperresponsiveness thus indicating the probable involvement of the src gene product in this phenomenon. The duration of the isoproterenol-induced cyclic AMP elevation was longer in the transformed than in the untransformed cells; transformed cells, unlike untransformed cells, required at least 120 min before full desensitization became established. Membranes prepared from transformed cells specifically bound more than 5 times the quantity of the beta-adrenergic radiolabeled antagonist (-)3H dihydroalprenolol and 125I-iodocyanopindolol compared to the untransformed cell membranes. Thus, it appears that major differences between the transformed and normal phenotypes reside in the concentration of membrane beta-adrenergic receptors and the inability of RSV-transformed cells to self-limit their response to specific external stimuli. PMID- 6284772 TI - Enzymatic modification of the surface carbohydrates of Friend erythroleukemic cells. AB - Surface carbohydrates of Friend erythroleukemic-cells were modified by treatment with the exoglycosidases, alpha-galactosidase, beta-galactosidase, and neuraminidase without affecting cell growth and viability either in the presence of absence of 1.8% DMSO as inducer. When cells were incubated with a combination of alpha-galactosidase and neuraminidase and then induced, they showed an increased rate of differentiation as measured by the formation of benzidine positive cells. These enzymes used singly, or beta-galactosidase treatment alone, or in combination with neuraminidase, did not change the rate of differentiation. Cell-surface labeling and electrophoretic separation of the glycoconjugates revealed that two regions of approximate molecular weights of 195,000 and 185,000 were neuraminidase-sensitive and one other of molecular weight of about 75,000 was sensitive to alpha-galactosidase. Both untreated and the combined alpha galactosidase, neuraminidase-modified cells exhibited the same rate of uptake of carbon-14 DMSO, ruling out the possibility that the observed increased rate of differentiation was due to faster penetration of DMSO into enzyme-treated cells. On the other hand, the decrease in the rate of uptake of rubidium-86, an analogue of K+, by treated-induced cells was significantly enhanced over that observed with untreated-induced cells, suggesting that alpha-galactosidase plus neuraminidase modification of the cell surface was affecting at least one of the early events occurring in the Friend erythroleukemic cell differentiation program. PMID- 6284771 TI - Relation between sodium-coupled amino acid and sugar transport and sodium/potassium pump activity in isolated intestinal epithelial cells. AB - Cells isolated from the epithelium of the small intestine are used to study the relationship between amino acid or sugar-coupled sodium transport and potassium uptake through the sodium/potassium pump. Potassium influx is a saturable function of the external potassium concentration. Uptake in the presence of ouabain, a specific pump inhibitor, is greatly reduced. This remaining influx is linearly related to the concentration up to 6 mM potassium. Sugars and amino acids are actively accumulated by the intestinal cells. Their transport is accompanied by an initial extra influx of sodium. Although cells seem to regulate their internal sodium concentrations, this is not accompanied with a concomitant increase in potassium uptake through the pump. Thus L-alanine, 3-0-methyl-D glucoside, and alpha-methyl-D-glucoside all fail to increase the rate of ouabain sensitive potassium uptake. A very high coupling ratio of sodium efflux to potassium influx through the pump would be a likely explanation of the present results though they cannot be regarded as conclusive. PMID- 6284774 TI - Semi-quantitative thin-layer mass-screening detection of 11-nor-delta 9 tetrahydrocannabinol-9-carboxylic acid in human urine. PMID- 6284773 TI - Does endogenous norepinephrine regulate potassium homeostasis and metabolism in rat cerebral cortex? AB - The role of endogenous cerebral norepinephrine (NE) as a modulator of transmembrane cation transport and energy metabolism was evaluated by monitoring extracellular potassium ion activity ([K+[o) in vivo and by measuring cortical Na+,K+-ATPase activity and oxygen consumption in vitro. Ipsilateral cortical NE was depleted by unilateral 6-hydroxydopamine (6-OHDA) lesions of the locus ceruleus (LC). The contralateral cortex was used for control measurements. NE depletion had no effect on resting levels of cortical [K+[o or on the rate of K+ removal from the extracellular space following direct cortical stimulation. There was also no effect of NE depletion on Na+,K+-ATPase activity in cortical homogenates nor on oxygen consumption of cortical slices over a wide range of K+ concentrations. These results indicate tht central NE depletion does not influence movements of cortical K+ either directly through an influence on Na+,K+ ATPase activity or indirectly through effects on oxidative metabolism. It is probable, therefore, that previously described effects of NE on cortical oxidative metabolism are mediated through changes in cerebral perfusion and/or modification of substrate availability in vivo. PMID- 6284775 TI - Gas chromatographic and mass spectrometric studies on the metabolism and pharmacokinetics of delta 1-tetrahydrocannabinol in the rabbit. AB - Gas chromatography-mass spectrometry has been used to investigate the in vivo hepatic metabolism of delta 1-tetrahydrocannabinol (delta 1-THC) in the New Zealand white rabbit. Sixteen metabolites were identified and shown to be present in different relative amounts compared with the hepatic metabolites of delta 1 THC produced by other species. The metabolic profile was also different from that reported from rabbit urine particularly with regard to the lower relative concentrations of acidic metabolites in the liver. The pharmacokinetics of delta 1-THC has been studied in the rabbit using the recently developed GC-MS method based on metastable ion monitoring. This revealed a terminal plasma delta 1-THC half life ranging from 34.16 to 59.30 h (mean 46.75 h) after a single dose and THC fat/plasma ratio of 10(3)-10(4):1. PMID- 6284776 TI - Determination of 5'-nucleotidase activity in human erythrocytes and plasma using high-performance liquid chromatography. AB - A method is described for the determination of 5'-nucleotidase activity in human erythrocytes and plasma. Using reversed-phase high-performance liquid chromatography; the product (uridine) was separated from the substrate (uridine 5'-monophosphate) in less than 4 min. The activity determined closely agreed with that determined by the conventional method, in which the inorganic phosphate released is measured. The present method eliminates the need for dialysis of enzyme solution prior to the assay, and offers several advantages over other assay methods, including high sensitivity. PMID- 6284777 TI - Use of high-performance liquid chromatography-mass spectrometry for the study of the metabolism of ranitidine in man. AB - High-performance liquid chromatography has been used for the qualitative analysis of metabolites and the quantitative analysis of ranitidine in the urine from subjects given oral and intravenous doses of ranitidine. Ranitidine, ranitidine-N oxide, ranitidine-S-oxide and desmethylranitidine were identified in extracts of the urine obtained by the XAD-methanol procedure. A selected-ion monitoring technique, using [2H3]ranitidine as the internal standard, was used to determine ranitidine. A normal-phase system consisting of methanol-propan-2-ol-5 M ammonium acetate (50:50:1) was used, and because of this the volume of urine which could be injected on-column without deterioration of the chromatography was limited to 10 microliters. This limited the sensitivity of the method to 1.0 microgram of ranitidine per millilitre of urine. PMID- 6284778 TI - Adrenocorticotropin-independent hypercortisolemia and testicular tumors in a patient with a pituitary tumor and gigantism. PMID- 6284779 TI - Characterization of the somatomedin-C/insulin-like growth factor I (SM-C/IGF-I) receptor on cultured human fibroblast monolayers: regulation of receptor concentrations by SM-C/IGF-I and insulin. PMID- 6284780 TI - Synthesis and glycosylation of proopiomelanocortin by a Cushing tumor. AB - ACTH derives from a larger protein, proopiomelanocortin (POMC), which is also the precursor to lipotropin, endorphin, melanotropin, and other peptides. The sequence of steps by which POMC is processed to ACTH has been delineated in detail in cultured mouse tumor cells, but little is known about these steps in human pituitary adenomas. Synthesis and glycosylation of POMC were studied in a pituitary adenoma causing Cushing disease and in adjacent tissue by incubating intact tissue explants in medium containing [35S]methionine or [3H]glucosamine. Labeled tissue was analyzed by two-dimensional gel electrophoresis, and a spot cut from these gels was analyzed by paper electrophoresis of tryptic digests. The location of the spot and the patterns of its [35S]methionine-labeled tryptic fragments resembled those found previously in other ACTH-secreting tissues and hence this spot appears to be POMC. Both the tumor and the adjacent pituitary produced this POMC, although tumor production of POMC was greater. The electrophoretic patterns of the putative POMC tryptic peptides labeled with [35S]methionine from this tumor are similar to the patterns of POMC tryptic peptides from a Nelson tumor and an ectopic ACTH- producing tumor when analyzed by the same techniques. This suggests that the excess ACTH in all three diseases arises from the same precursor POMC molecule. Two-dimensional gel electrophoresis of [3H]glucosamine-labeled tumor suggests that the first proteolytic cleavage occurs between the alpha ACTH-(1-39) and beta-lipotropin sequences and that a subsequent cleavage occurs between the N-terminal region of POMC and ACTH. Paper electrophoresis of [3H]glucosamine-labeled tryptic glycopeptides reveals at least three radioactive peaks, suggesting that the two known glycosylation sites in human POMC may be variably glycosylated. PMID- 6284781 TI - Nuclear binding of [125I]triiodothyronine in dispersed cultured skin fibroblasts from patients with resistance to thyroid hormone. PMID- 6284782 TI - Down-regulation of insulin receptors in postnecrotic cirrhosis of liver. AB - Eighteen patients with postnecrotic cirrhosis of liver, including three patients who had had a portosystemic shunt operation, and 19 normal controls were studied. The tests performed included monocyte insulin receptor assay, iv glucose tolerance test, glucagon test, and insulin tolerance test. Insulin resistance was documented by the presence of fasting hyperglycemia and glucose intolerance together with hyperinsulinemia as well as resistance to exogenous insulin. The binding of [125I]insulin to monocyte insulin receptors was significantly decreased in cirrhotic patients compared with that in controls (P less than 0.02), and this was due to a significant decrease in the high affinity association constant (P less than 0.005). There was a significant negative correlation between the fasting insulin level and maximum [125I]insulin binding in cirrhotic patients (r = -0.8; P less than 0.02). Cirrhotics that had had a shunt operation showed a higher fasting insulin level, a greater insulin resistance, and a smaller maximum [125I]insulin binding to insulin receptors than those without shunt. All of these findings suggested a down-regulatory effect of hyperinsulinemia on the monocyte insulin receptors. An impaired glycemic response to glucagon was also found in cirrhotics, the exact mechanism of which remains to be elucidated. However, as the increases in plasma cAMP after glucagon were similar in cirrhotics and controls, the fault apparently did not lie in the glucagon receptor. PMID- 6284783 TI - Dynamics of adrenocorticotropin (ACTH) secretion in cyclic Cushing's syndrome: evidence for more than one abnormal ACTH biorhythm. AB - We have studied a 57-yr-old woman with cyclic Cushing's syndrome of apparent pituitary origin who had a predominant cycle of 2-6 days. The patient also demonstrated an abnormal circadian rhythm, with afternoon peaks of plasma ACTH and plasma cortisol. In addition to these abnormal biorhythms, Fourier analysis showed what appeared to be a separate 35-day cycle. After 35 days of consecutive urinary free cortisol measurement, the patient was given cyproheptadine. During therapy with this agent, the urinary free cortisol levels fell dramatically, but cyclic secretion continued, albeit with a diminished amplitude. During general anesthesia for a bilateral adrenalectomy, there was a striking increase in the plasma ACTH level, and the ACTH concentration remained high in both the immediate and late postoperative periods. These observations indicated that stress could overcome cyclic ACTH secretion and that cortisol exerted feedback suppression on ACTH secretion. Although this is the predictable response for classic pituitary dependent Cushing's syndrome, it is of interest in cyclic Cushing's syndrome, since previous studies of this entity have implied that cortisol secretion is independent of stimulation or feedback. PMID- 6284784 TI - Properties of human growth hormone polypeptides: purified from pituitary extracts and synthesized in monkey kidney cells and bacteria. AB - Several forms of human GH (hGH) have been elucidated by extraction of human pituitary and recombinant DNA techniques. In the present study we have characterized five of these hGH polypeptides by gel filtration, RIA, and radioreceptor assays. These include extractable pituitary hGH and its naturally occurring 20K variant. Two hGH polypeptides were produced from naturally occurring human genes in simian kidney cells (SV-hGH 1 and 2) and another preparation was produced from a partially synthesized gene in bacteria (E. coli hGH). As predicted from their known DNA sequences, naturally occurring pituitary hGH, SV-hGH 1, and E. coli-hGH migrated as a single peak on Sephadex G-100 column and had the same immunological and receptor-binding properties. By contrast, SV hGH 2 (14 dispersed amino acid substitutions) and the 20K variant (amino acid residues 32-46 deleted from hGH) contained more higher molecular weight components and had diminished immunological and receptor-binding potency. SV-hGH 2 differed from the 20K variant by having even lower immunological potency and containing more of the higher molecular weight component. These variant forms of hGH may provide an explanation for the heterogeneity of both pituitary and plasma hGH. PMID- 6284785 TI - Steroid replacement in Addison's disease and in subjects adrenalectomized for Cushing's disease: comparison of various glucocorticoids. PMID- 6284786 TI - Preparation of 125I-labeled receptor-purified Graves' immunoglobulins: properties of their binding to human thyroid membranes. PMID- 6284787 TI - Central deficiency of beta-endorphin in alcohol addicts. AB - Alcohol addiction may induce its dependence through a mechanism involving opiate receptors and opioid peptides. For these reasons, we measured ACTH, beta lipotropin, and beta-endorphin in the plasma and cerebrospinal fluid (CSF) of 29 alcohol addicts and compared these values with those found in 8 normal volunteers. Although no significant differences existed in peripheral concentrations of the 3 peptides, alcohol addicts had beta-endorphin levels in CSF (mean +/- SE, 29.4 +/- 4.5 fmol/ml) that were 3-fold lower than those of the controls (98.4 +/- 10.5 fmol/ml; P less than 0.001) and ACTH levels 4 times higher than control values (30.0 +/- 1.8 vs. 7.4 +/- 1.1 fmol/ml in controls; P less than 0.001), while no difference was found in beta-lipotropin levels. These results suggest that alcohol addiction is associated with a marked alteration in the CSF content of proopiocortin-related peptides which may play a role in the alcohol-seeking behavior typical of the syndrome. PMID- 6284788 TI - The role of the human major histocompatibility complex in cytotoxic T-cell responses to virus-infected cells. AB - The human major histocompatibility complex (HLA) has been demonstrated to play two roles in the generation and expression of cytotoxic T-lymphocyte responses to virus-infected cells: (1) cytotoxic T cells can only recognize viral antigens in conjunction with antigens encoded by HLA-A and -B genes; and (2) HLA-linked genes may control the capacity to generate T-cell responses to a given virus or to virus in conjunction with particular self HLA-A and -B antigens. Analysis of T cell responses generated in vivo to Epstein-Barr virus suggests that human T cells may recognize virus in conjunction with antigens other than the class I HLA polymorphic specificities. PMID- 6284790 TI - [Studies on pathogenesis of cerebral edema due to experimental cerebral hemorrhage. An approach through energy metabolism (author's transl)]. PMID- 6284789 TI - Thyroid-stimulating hormone (TSH) receptor antibodies and antibodies stimulating adenyl cyclase in relatives from two families with a high prevalence of Graves' hyperthyroidism: a ten-year follow-up study. AB - Thyroid-stimulating hormone (TSH) receptor antibodies and antibodies stimulating adenyl cyclase were measured in 47 relatives of patients with Graves' hyperthyroidism from two families with a high prevalence of the disease, in whom bioassays for the long-acting thyroid stimulator (LATS) had been performed 10 years earlier. Tests were also carried out in six propositi from the two families and age- and sex-matched normal subjects from six families. There had been no new cases of hyperthyroidism since the first study, although one subject was clinically and biochemically hyperthyroid at the time of study and two more were biochemically borderline hyperthyroid but clinically euthyroid. Levels of serum T4 thyrotropin, and percentage T3 resin uptake and free thyroxine indices were similar for relatives and normal subjects, although the mean serum T3 level for relatives was significantly greater than that for the normal subjects. Antibodies were not detected by either assay in any relative. Significant titers of antithyroglobulin antibodies were demonstrated in 4 of 44 relatives but in none of 46 normals tested, while thyroid cytoplasmic antibodies were detected in 8 of 44 relatives and 3 of 45 normals. The mean serum IgG for Graves' relatives was significantly greater than that for the normals, although the mean IgM and IgA levels for the two groups were not significantly different. PMID- 6284791 TI - Evaluation and reporting of enzyme immunoassay determinations of antibody to herpes simplex virus in sera and cerebrospinal fluid. AB - Several methods for evaluating and reporting enzyme immunoassay (EIA) determinations of antibody to herpes simplex virus derived from one dilution of single serum samples were studied. An EIA ratio method for serological evidence of current infection from paired serum samples was also evaluated. Optical density (OD) of the reaction at a 1:100 serum dilution and estimated titers obtained by reference of the OD of the serum dilution to a standard curve were compared to the corresponding plotted EIA titer obtained by titration to endpoint. Neither the OD per se nor the estimated titer was completely predictive of the plotted titer (correlation coefficient [r] of 0.824 and 0.817, respectively), and they provided only a semiquantitative measurement of antibody concentration. For an antibody status report, however, OD would be sufficient if related to the cutoff value as an EIA index (OD of sample divided by cutoff OD for positive specimens). The OD of the EIA reaction at a single dilution (1:5) of cerebrospinal fluid, on the other hand, correlated quite well with the titer obtained by titration (r = 0.950). For serological diagnosis of current infection, the OD ratio of convalescence-phase/acute-phase sera was determined at several dilutions. A ratio of greater than or equal to 1.54 was calculated as a reliable index for a significant rise in antibody concentration and compatible with current infection. By determining the convalescent-phase/acute-phase serum ratio at two dilutions, 1:100 and 1:1,000, the EIA ratio method appeared to be a sensitive as or more sensitive than, complement fixation in diagnosing current infection. PMID- 6284792 TI - Comparison of guinea pig embryo cells, rabbit kidney cells, and human embryonic lung fibroblast cell strains for isolation of herpes simplex virus. AB - With the recognition of the importance and the prevalence of herpes simplex virus infections and the availability of effective chemotherapy, there has come a demand for rapid, sensitive, and accurate diagnosis. Virus isolation in cell culture remains the most widely employed diagnostic method, and the cell cultures used vary greatly from laboratory to laboratory. Therefore, a study was undertaken to compare several commonly employed human fibroblast cell strains and two nonhuman cells, primary rabbit kidney and guinea pig embryo cells, for the primary isolation of this virus. Our experience in the isolation of herpes simplex virus in the clinical laboratory with these cell systems was evaluated, and reisolations from clinical specimens were performed. We found that guinea pig embryo and rabbit kidney cell cultures were equally sensitive for the isolation of herpes simplex virus from clinical specimens and were superior to the human embryonic lung fibroblast cell strains tested in terms of both sensitivity and rapidity of appearance of cytopathic effect. The use of guinea pig embryo cells for the isolation of this virus from clinical specimens has not been previously reported. PMID- 6284793 TI - Comparison of an enzyme immunoassay with electron microscopic procedures for detecting rotavirus. AB - The sensitivity and specificity of an enzyme immunoassay (Rotazyme), an ongrid immunoelectron microscopy procedure, and conventional negative stain electron microscopic techniques were compared. By using partially purified human rotavirus and simian rotavirus (SA-11) of known particle concentration, the enzyme immunoassay was essentially equivalent to the immunoelectron microscopic procedure and significantly more sensitive than conventional electron microscopic techniques. The level of sensitivity was approximately 10(6) particles per ml for simian rotavirus SA-11 and 10(7) particles per ml for human rotavirus. In an evaluation of 455 clinical samples by these techniques, a sensitivity of 98% and specificity of 92% were demonstrated. Samples negative by the immunoelectron microscopic procedure and positive by enzyme immunoassay could be confirmed by a blocking assay. PMID- 6284794 TI - Infectivity of Chlamydia trachomatis in tissue culture with newborn calf serum. AB - By using representative strains of Chlamydia trachomatis serotypes, the efficacy of substituting fetal bovine serum with newborn calf serum for the isolation of this organism in tissue culture was examined. The criteria used were the quality and quantity of the iodine-stained cytoplasmic inclusions and the characteristics of the McCoy cells. Complete substitution of fetal bovine serum with newborn calf serum produced a detrimental change in the quality of the cytoplasmic inclusions and a dramatic decline in the inclusion count (P less than 0.001) with all of the chlamydial strains tested. There appeared to be no significant alteration in the characteristics of the McCoy cells. It is recommended from this preliminary investigation that newborn calf serum should not be used for the isolation of C. trachomatis in tissue culture. PMID- 6284795 TI - Immunological model of epilepsy. Epileptiform activity induced by fragments of antibody to GM1 ganglioside. AB - Antibodies to GM1 ganglioside injected into the sensorimotor cortex of the rat induce recurrent epileptiform activity. We now find that the divalent F(ab')2 and monovalent Fab' fragments derived from antiganglioside IgG molecules are able to induce epileptiform seizures. This result supports the view that the binding of antibodies to ganglioside receptors in the synaptic membrane is sufficient in itself to initiate changes in membrane processes which lead to epileptiform spiking. These changes do not appear to be dependent on linking of ganglioside receptors or on the presence of serum factors such as complement. PMID- 6284796 TI - Human fetal brain antigen expression common to tumors of neuroectodermal tissue origin. AB - The antigenic relationship between human tumors of neuroectodermal origin and fetal brain were further investigated by characterization of two hybridoma antibodies derived from a fusion of P3-NS1/1-Ag 4-1 (NSI) myeloma cells and splenocytes hyperimmunized to second trimester human fetal brain homogenate. Monoclonal antibodies (MAs) 1H8cl 2 and 1H8cl 3 were analyzed by cell surface radioimmunoassay (CS-RIA), quantitative absorption, indirect immunofluorescence, and peroxidase-antiperoxidase (PAP) immunohistology. MA 1H8cl 3 is the more broadly reactive, binding to 9/14 glioblastoma (GBM), 2/3 neuroblastoma, 1/2 melanoma, and 1 medulloblastoma cell line(s) by CS-RIA analysis, and to 12/15 GBM, fetal brain, spleen, and liver, and adult spleen by PAP analysis. MA 1H8cl 2 is more restricted, binding to 7/14 GBM, 2/3 neuroblastoma, 1 medulloblastoma, and 2/3 fetal skin fibroblast cell line(s) by CS-RIA, and to 9/15 GBM and fetal brain and spleen by PAP analysis. Control non-central nervous system tumors and normal adult tissue including brain, thymus, lymph node, liver, kidney, lung, skin, and pancreas, were unreactive with both 1H8cl 2 and 1H8cl 3 by CS-RIA, PAP, and absorption analysis. The data presented here establish the unique nature of the detected antigenic specificities as compared to previously described oncofetal and onconeural antigens, and define two immune reagents which are operationally specific for tumors of neuroectodermal origin within the adult central nervous system. PMID- 6284797 TI - Consequences of potassium recycling in the renal medulla. Effects of ion transport by the medullary thick ascending limb of Henle's loop. AB - The consequences of K recycling and accumulation in the renal medulla were examined by measuring the effect of elevated K concentration on ion transport by the medullary thick ascending limb of Henle's loop. Perfused and bathed in vitro, thick limbs from both mouse and rabbit displayed a graded, reversible reduction of transepithelial voltage after increasing K concentration from 5 to 10, 15, or 25 mM. The effect was reproducible whether osmolality was 328 or 445 mosmol/kg H2O, and whether K replaced Na or choline. Net chloride absorption and transepithelial voltage were reduced by almost 90% when ambient K concentration was 25 mM. When either lumen or bath K was increased to 25 mM, net Na absorption was reduced. There was spontaneous net K absorption when perfusate and bath K concentration was 5 mM. Analysis of transepithelial K transfer after imposition of chemical gradients demonstrated rectification in the absorptive direction. Absorption of K by this segment provides a means to maintain high medullary interstitial concentration. Accumulation of K in the outer medulla, by reducing NaCl absorption, would increase volume flow through the loop of Henle and increase Na and water delivery to the distal nephron. K recycling thus might provide optimum conditions for K secretion by the distal nephron. PMID- 6284798 TI - Characterization of the platelet membrane glycoprotein abnormalities in Bernard Soulier syndrome and comparison with normal by surface-labeling techniques and high-resolution two-dimensional gel electrophoresis. AB - The platelets from three patients with Bernard-Soulier syndrome have been analyzed by surface-labeling coupled with two-dimensional gel electrophoresis and compared with normals. As well as the previously described absence or deficiency in glycoprotein (GP) Ib(alpha) it could be shown that GP Ib beta and an additional low molecular weight glycoprotein GP17 were not detectable using carbohydrate-labeling methods or deficient to the same extent as the GPIb alpha subunit. In addition, the thrombin cleavable glycoprotein could not be detected using carbohydrate-labeling methods in two patients and was deficient in a third. This finding was confirmed in a fourth patient by one-dimensional gel electrophoresis. Thus, the changes in the membrane of Bernard-Soulier platelets are more complex than previously thought. PMID- 6284799 TI - Abnormal renin short feedback loop in essential hypertension is reversible with converting enzyme inhibition. AB - The suppression of renin release by angiotensin II (AII) (the so-called short feedback loop) is blunted in essential hypertension. To determine whether this abnormality is reversible, renin release was assessed in sodium-restricted essential hypertensives and normal controls: (a) during the administration of captopril for varying intervals and (b) following the infusion of graded doses of AII (0.3-3 ng/kg per min) before and after plasma levels of AII had been chronically reduced with captopril (25-50 mg every 6 h) for 70 h. In control subjects, the maximal increment above control in plasma renin activity (PRA) after a single dose of captopril (11.9+/-3 ng/ml per h) was significantly (P < 0.02) greater than in hypertensives (8.1+/-1.7 ng/ml per h) despite similar reductions in AII levels and significantly greater decrements in diastolic blood pressure in the hypertensives. When captopril was continued for 70 h, the PRA increments above base line in hypertensive subjects (11.4+/-2.9 ng/ml per h) rose to levels seen in the controls (11+/-2.6 ng/ml per h); there were no significant differences in the AII or diastolic blood pressure decrements between the two groups. Compared with normotensive subjects, AII failed to suppress renin release in hypertensive subjects despite significantly greater diastolic blood increments and comparable AII levels achieved at each AII dose. After captopril treatment, AII now produced significant declines in PRA in the hypertensives; moreover, comparing declines pre- and postcaptopril, greater PRA decrements were seen either at comparable rises in levels of AII or diastolic blood pressure. Finally, the suppression of PRA by AII postcaptopril in hypertensives was now indistinguishable from that seen in normal controls. Thus, the impaired regulation of renin by AII is reversible with prolonged captopril treatment, suggesting that this abnormality is not due to a fixed structural defect but to a reversible lesion. PMID- 6284800 TI - Lung cell oxidant injury. Enhancement of polymorphonuclear leukocyte-mediated cytotoxicity in lung cells exposed to sustained in vitro hyperoxia. AB - The oxidant damage of lung tissue during in vivo hyperoxic exposure appears to be amplified by neutrophils that release toxic amounts of oxygen metabolites. In our studies cloned lung epithelial cells (L2 cells), lung fibroblasts, and pulmonary artery endothelial cells were cultured under either ambient (Po(2) approximately 140 torr) or hyperoxic (Po(2) approximately 630 torr) conditions for 48 h (24 h for endothelial cells). After cultivation, phorbol myristate acetate- or opsonized zymosan-stimulated neutrophils were added to the cultivated monolayers for 4 h, and lung cell damage was quantitated using (51)Cr release as an index. The data show that stimulated neutrophils are able to injure the three lung cell lines tested, with endothelial cells being highly susceptible to this injury and L2 cells being slightly more susceptible than lung fibroblasts. The studies also demonstrate that all three lung cell lines exposed to sustained hyperoxia are more susceptible to neutrophil-mediated cytotoxicity than their time-matched air controls. Hydrogen peroxide was the main toxic oxygen metabolite because catalase (2,500 U/ml) completely protected the target cells. Equivalent quantities of hydrogen peroxide generated by glucose oxidase instead of by neutrophils gave a similar degree of target cell injury. Superoxide dismutase at high concentrations (250 mug/ml) provided some protection. Other systems that detoxify oxygen metabolites were without protective effect. These findings indicate that the increase in susceptibility of lung cells to neutrophil-mediated oxidant damage is a toxic effect of hyperoxia on lung cells. This specific manifestation of oxygen damage provides insight into the integration between primary mechanisms (oxygen exposure) and secondary mechanisms (release of oxygen metabolites by neutrophils) with respect to the cellular basis for pulmonary oxygen toxicity. PMID- 6284803 TI - Hepatitis B and A virus antibodies in alcoholic steatosis and cirrhosis. AB - Sera from 74 alcoholics with cirrhosis and 63 alcoholics with steatosis were tested for antibody to hepatitis B surface antigen, to hepatitis B core antigen, and to hepatitis A virus by radioimmunoassay or enzyme-linked immunosorbent assay. No significant difference between the two groups of alcoholics could be found concerning the prevalence of these antibodies. The total group of patients had antibody to hepatitis B surface antigen or hepatitis B core antigen, or both, significantly (p less than 0.001) more often (26%) than sex- and age-matched controls (4%). No significant difference was found between patients and controls concerning the prevalence of antibody to hepatitis A virus (46% v 40%). In patients with cirrhosis, no correlation between wedged hepatic vein pressure or wedged-to-free hepatic vein pressure and any of the viral antibodies could be established. The present results suggest that hepatitis B virus does not play a major role in the progression of alcoholic liver disease, but longitudinal studies are needed to solve this problem. The reason for the increased prevalence of antibodies to hepatitis B virus in these patients is unknown. PMID- 6284802 TI - Mineralocorticoid and glucocorticoid effects on 31,000- and 29,000-dalton proopiomelanocortin in rat anterior pituitary and neurointermediate lobe. AB - The effects of adrenal steroids on proopiomelanocortin (POMC) levels in rat pituitary have been studied by two-dimensional gel electrophoresis. In intact rats the relative abundance of POMC was much higher in the neurointermediate lobe (N-IL) than in anterior pituitary (AP); in both tissues the predominant species appeared to be of 29,000-dalton (29K) molecular mass, with lesser amounts of a 31K form. In both tissues, the 31K and 29K forms showed multiple spots, consistent with different degrees of sialoglycosylation. Adrenalectomy was followed by a marked increase in AP levels of POMC, and a marked decrease in N-IL levels. In adrenalectomized rats, dexamethasone administration did not affect N IL levels of POMC, but suppressed 35S incorporation into POMC in AP in a dose related manner; deoxycorticosterone showed minimal effects on AP levels of POMC, but progressively elevated N-IL levels; 9 alpha fluorocortisol (9 alpha fF) progressively both suppressed AP levels, and raised N-IL levels of POMC. Estimation of immunoreactive (ir) ACTH and ir-beta-endorphin in parallel samples showed an elevation of N-IL levels in response to mineralocorticoids (deoxycorticosterone, 9 alpha fF), and a paradoxical elevation of AP levels in response to glucocorticoids (dexamethasone, 9 alpha fF) compared with oil injected adrenalectomized controls. We conclude (a) that glucocorticoids suppress the secretion of ir-ACTH and ir-beta-endorphin to a greater extent than they inhibit the synthesis of POMC; (b) that mineralocorticoids specifically elevate the N-IL levels of both POMC and its immunoreactive product (beta-endorphin). PMID- 6284801 TI - Thromboxane mediation of cardiopulmonary effects of embolism. AB - Humoral factors released from platelets during pulmonary embolism may be the cause of several attendant cardiopulmonary abnormalities. This study examines the role of thromboxanes (Tx) after experimental embolism induced with 0.5 g/kg autologous clot in four groups of five dogs: (a) untreated embolized controls; (b) pretreatment with the Tx synthetase inhibitor, imidazole 25 mg/kg . h i.v., starting 30 min before embolization; (c) pretreatment with the cyclooxygenase inhibitor indomethacin, 5 mg/kg, 12 h per os and 1 mg/kg, 1 h i.v. before the experiment; (d) treatment with prostacyclin (PGI(2)) 100 etag/kg . min i.v. for 1 h, 1 h after embolization. Within 30 min, embolization led to increases of 6-keto PGF(1alpha), the stable hydrolysis product of PGI(2), from 0.11+/-0.08 etag/ml (mean+/-SD) to 0.33+/-0.10 etag/ml (P < 0.005) and TxB(2), the stable product of TxA(2), from 0.10+/-0.04 etag/ml to 0.38+/-0.06 etag/ml (P < 0.001). Increases were observed in total dead space (V(D)/V(T)) from 0.46+/-0.03 to 0.61+/-0.08 (P < 0.025, physiologic shunting (Q(S)/Q(T)) from 16+/-4% to 38+/-9% (P < 0.01), pulmonary vascular resistance (PVR) from 2.27+/-0.59 mm Hg.min/liter to 9.21+/ 1.90 mm Hg.min/liter (P < 0.005) and mean pulmonary arterial pressure from 14+/-6 mm Hg to 34+/-1 mm Hg (P < 0.001). Cardiac index (CI) fell from 139+/-11 ml/kg.min to 95+/-17 ml/kg.min in 4 h (P < 0.025). Imidazole pretreatment prevented a rise of TxB(2), but not 6-keto-PGF(1alpha); indomethacin blocked both. Both agents maintained V(D)/V(T) at base line and limited increases in Q(S)/Q(T) and PVR. CI was higher after imidazole pretreatment compared with controls (P < 0.025). Indomethacin led to intermediate levels of CI. PGI(2) lowered TxB(2) (P < 0.025), V(D)/V(T) (P < 0.025), Q(S)/Q(T) (P < 0.025) and PVR (P < 0.05) within 30 min. During PGI(2) infusion, CI was higher than controls. Concentrations of TxB(2) correlated with V(D)/V(T), r = 0.79 and Q(S)/Q(T), r = 0.69 (P < 0.001). Treatment of three dogs with the imidazole derivative ketoconazole, 10 mg/kg IV, 30 min after 0.75 g/kg autologous clot resulted in a lowering of physiologic dead space, but no other improvement of cardiopulmonary function. These results show that a number of cardiopulmonary abnormalities induced by pulmonary embolism are related directly or indirectly to platelet secretions and that V(D)/V(T) is closely allied to TxA(2) levels. PMID- 6284804 TI - Idiopathic narcolepsy: effect of naloxone studied in one of three patients with unelevated beta-endorphin-like immunoreactivity plasma levels. PMID- 6284805 TI - The thalamo-cortical projection of the nucleus submedius in the cat. AB - The cortical projection of the nucleus submedius (Sm) was studied in the cat with the autoradiographic and horseradish peroxidase (HRP) methods. The results indicate that Sm projects topographically on to layer 3 of a distinct agranular cortical field that occupies the posterolateral gyrus proreus, the adjacent fundus of the rhinal sulcus, and the postero-ventral portion of the medial wall of the presylvian sulcus. This cortical field is denoted the ventrolateral orbital cortex (VLO), consonant with previous nomenclature in the rat (Krettek and Price, '77a). The more ventral part (VLO beta) is cytoarchitectonically distinct from the dorsal part (VLO alpha); the former receives input from the anterior part of Sm (Sma), while the latter receives input from the dorsal and ventral parts of Sm (Smd and Smv). A light input to superficial layer 1 of VLO probably also arises from Sm, and there may be an input to layers 5 and 6. The corticothalamic projection from VLO to Sm reciprocates the ipsilateral thalamocortical projection and also has a moderate contralateral component. A dense, subpial layer 1 input to VLO arises from cells of the ventromedial nucleus (VM) subjacent to Sm. The present experiments also indicate that clusters of cells in VM probably provide input to layer 3 of the cortex in the fundus of the presylvian sulcus, as well as area 6a beta in the lateral wall of the presylvian sulcus and the ventral bank of the cruciate sulcus. Results from the HRP experiments additionally indicate that VLO beta and the anteroventral (Smv) portion of VLO alpha are reciprocally connected with the ventral agranular insular cortex and the cingulate cortex, ipsilaterally, while the posterodorsal (Smd) portion of VLO alpha is instead connected wih specific portions of the somatosensory cortical areas bilaterally. All portions of VLO alpha appear to project to the ventrolateral periaqueductal gray. In light of the recent suggestion that Smd is involved with nociception (Craig and Burton, '81), the present results suggest that the related portion of VLO alpha may serve as a cortical representation for noxious stimuli. PMID- 6284807 TI - The effect of post-infection immunization on the severity of experimental visna. PMID- 6284808 TI - Effects of selenium and copper deficiency on neutrophil function in cattle. PMID- 6284809 TI - Gianotti-Crosti-like eruption associated with coxsackievirus A-16 infection. PMID- 6284806 TI - Selective uptake of lucifer yellow by retinal cells. AB - When turtle retinae were incubated with the fluorescent dye, lucifer yellow, in the absence of Ca2+, the dye was selectively accumulated by cell bodies located in the inner nuclear layer (INL). The morphological features of the labeled cells suggested that they were bipolar cells. Other fluorescent dyes, Procion yellow and Primulin, were also taken up by somata in the INL, in the absence of external Ca2+, although the identity of the labeled cells was uncertain. As with turtle retina, lucifer yellow was accumulated predominantly by cell bodies in the INL of goldfish, frog, and rat retinae. Lucifer yellow uptake appeared to be independent of synaptic activity since dark-adaptation or aspartate treatment of retinae did not alter the dye uptake. Further, retinae from dystrophic (RCS) rats showed uptake similar to that seen in normal rat retinae. After uptake, most of the dye was found intracellularly as patches or vacuoles in the somata of the labeled cells. Dye uptake was not inhibited by removal of Na+ from the incubation medium. Further, prior treatment with metabolic inhibitors, cyanide and iodoacetate, or cytochalasin B, did not block the dye uptake. PMID- 6284810 TI - Immunosuppression in a homosexual man with Kaposi's sarcoma. AB - The occurrence of Kaposi's sarcoma in young homosexual men is a recently reported condition. The same individuals are at risk for the development of Pneumocystis carinii pneumonia and other unusual infections. These associations suggest these persons are somehow immunocompromised. Evidence of current or prior cytomegalovirus (CMV) infection is seen in a high percentage of these and other homosexual men. CMV infections are known to induce alterations in the immunoregulatory suppressor and helper T lymphocyte populations. The CMV infection is suspect as the cause of immunosuppression in these individuals. We present a case of Kaposi's sarcoma in a homosexual men with CMV cultured from his urine and semen. He showed a marked increase in his suppressor/cytotoxic cell (OKT8-positive) population, as well as a marked decrease in his helper cell (OKT4 positive) population. Mitogen and antigen studies demonstrated absent or markedly diminished response both in vitro and in vivo. Pokeweed mitogen (PWM)-induced IgG synthesis appeared normal. This patient, as well as the majority of other reported patients with this disease, manifested the HLA-Dr5 phenotype. The immunosuppression in this patient and possibly other similar men appears to be mediated by abnormalities in the immunoregulatory T lymphocytes. PMID- 6284811 TI - Effects of hepatic denervation on the anorexic response to epinephrine, amphetamine, and lithium chloride: a behavioral identification of glucostatic afferents. AB - Intraperitoneal injections of epinephrine (20, 40, 80, and 160 microgram/kg) and amphetamine (.1, .2, and .4 mg/kg) were administered to rats with various forms of hepatic denervation. In Experiment 1, destruction of the esophageal trunks of the vagus attenuated epinephrine and amphetamine anorexia, but destruction of the hepatic vagus did not. In Experiment 2, rats with celiac ganglionectomy, subdiaphragmatic vagotomy, or the combined operation all exhibited decreased epinephrine anorexia to the same extent. However, ganglionectomized rats were less responsive to amphetamine anorexia than were vagotomized ones. Vagotomized rats were significantly more reactive to lithium chloride (10, 20, and 30 mg/kg) than were controls. These results suggest that the major component of hepatic metabolic afferent fibers travels from the liver, through the celiac ganglion, and into the esophageal vagal trunks where they ascend to the brain. The anorexic action of amphetamine appears to result from a centrally induced sympathetic action on the liver. PMID- 6284813 TI - Computed tomography of benign hepatic tumors. AB - Computed tomography (CT) was performed on five patients with focal benign liver lesions: two cases of focal nodular hyperplasia (FNH) and three cases of hepatic adenoma (HA). All five patients had low density lesions seen best on noncontrast scans. Focal nodular hyperplasia often presents as an asymptomatic, well circumscribed lesion with a central, stellate, fibrous scar. Hepatic adenoma, which is associated with the use of oral contraceptives, is a smooth bordered mass that exhibited foci of hemorrhage in two of our three cases. Both of these benign liver lesions enhance to a variable degree of following intravenous contrast medium administration. It is concluded that a specific diagnosis of FNH or HA may be suggested when appropriate CT findings are detected. PMID- 6284812 TI - Cortical substrates of taste aversion learning: dorsal prepiriform (insular) lesions disrupt taste aversion learning. AB - The functional relation between restricted damage to ventral primary somatosensory neocortex and the ability of rats to acquire conditioned taste aversion (CTA( was examined by a combination of behavioral and neurohistological techniques. Lesions confined exclusively to the established gustatory neocortex (GN) did not disrupt CTA acquisition, nor did lesions confined to suprarhinal cortical areas ventral to the GN. Lesions that encroached on dorsal prepiriform and insular cortices produced CTA acquisition deficits and damaged a large proportion of efferent projections to the prefrontal and precentral neocortex. In a second experiment, lesions of dorsal prepiriform and insular cortices did not modify taste preference-aversion threshold to any of the four taste modalities. It is concluded tha ventral somatosensory neocortical fields, including the established GN, do not mediate CTA acquisition and that rhinal cortices ventral and posterior to the GN are preferentially involved in associative learning for tastes and illness. PMID- 6284814 TI - Brain tumor imaging by positron emission computed tomography using 11C-labeled amino acids. PMID- 6284815 TI - Effect of dexamethasone treatment on peritumoral brain edema: evaluation by computed tomography. AB - In a prospective study, the effect of dexamethasone treatment on brain edema was evaluated by computed tomography (CT) in 14 patients with intracranial neoplasm. A CT study was made immediately prior to the beginning of the treatment and at various intervals for 8 to 19 days thereafter. The volume and attenuation of the edema were measured and related to the time after the start of treatment. In five cases of meningioma, only minor changes were shown; the edema volume either decreased or increased slightly or did not change at all. In four gliomas, a substantial decrease was noted in the edema volume in two cases and a smaller decrease was observed in two cases. In three metastases, a linear decrease in edema volume was noted during the entire follow-up, reducing it to one-fourth of the initial volume after 2 weeks of treatment. In one case of acoustic neuroma and in one undiagnosed lesion, a marked decrease in edema volume was also noted. The attenuation of the remaining edema was constant during the entire treatment time. PMID- 6284816 TI - Pigmented hidroacanthoma simplex with porocarcinoma. Light and electron microscopic study of a case. AB - A case of pigmented hidroacanthoma simplex showing malignant transformation into porocarcinoma is reported. Although no intracellular duct formation could be observed as in benign tumors, ultrastructurally the tumor cells showed characteristics similar to those of eccrine poroma. Many melanocytes were seen dispersed within the tumoral nests. The melanocyte-keratinocyte relationship was found similar to that occurring in melanoacanthoma. Porocarcinoma cells showed ultrastructural features similar to those of benign cells. PMID- 6284818 TI - Influence of parathyroid status of rats on renal tubular handling of adenosine 3'5'-monophosphate: a micropuncture study. AB - In order to correlate cyclic AMP handling by the nephron to the parathyroid status, clearance and micropuncture experiments were performed in rats with intact parathyroid glands, or immediately after parathyroidectomy, or six days after parathyroidectomy. In intact animals cyclic AMP urinary excretion was about twice the filtered load and the tubular addition of the nucleotide was achieved at the end of the accessible proximal tubule. In acutely parathyroidectomized rats cyclic AMP urinary excretion was not different from the filtered load and no proximal tubular addition was detected at the late accessible proximal tubule. In chronically parathyroidectomized animals urinary excretion of cyclic AMP was not different from the filtered load, nevertheless a proximal tubular addition of the nucleotide was observed, similar in magnitude to that of intact rats. The data afford a direct evidence that the convoluted proximal tubule is the major site of cyclic AMP tubular addition, confirm that this addition disappears immediately after parathyroidectomy, but indicate that it re-occurs in chronic parathyroidectomy. PMID- 6284817 TI - Membrane milieu is regarded as the native environment of the cyclic AMP degrading enzyme cluster. AB - The phenomenon of kinetic advantage of nucleoside formation from cyclic AMP, via the intermediate 5'AMP has been observed in the microsomal fraction after subcellular fractionation of beef adrenal cortex tissue. It was explained by the existence of a multienzyme sequence previously evidenced [H. Wombacher, 1982, Arch. Biochem. Biophys. 201, 8-19]. In the present study a similar enzyme cluster was prepared from the soluble fraction of the cell homogenate after two steps of gel-chromatography. An elusive channeling of cyclic AMP degradation could be disclosed. The time course reaction of cyclic AMP degradation to the nucleosides, adenosine and inosine, via 5'AMP as an intermediate compared with the time course reaction of 5'AMP hydrolysis to the nucleosides, adenosine and inosine, under otherwise identical conditions showed that the nucleoside formation from cyclic AMP was faster after the lag phase of the reaction sequence. This kinetic advantage effect, however, was much less pronounced than to be seen in the membrane-bound multienzyme sequence. For an analysis of the influence of the environmental conditions on the activity of both enzyme cluster forms they were treated by chaotropic agents, detergents and ultrasonic power. Common to all results was: the activity of the membrane-bound enzyme cluster is highly stable in comparison with the soluble form. On basis of these and previous findings a hypothesis is suggested explaining the similarities between the membrane-bound enzyme cluster and the soluble form. Thus, the soluble enzyme cluster form is considered a partially preserved form of the membrane-bound form arisen from the cell homogenization process and/or vice versa the soluble form might present a pro-form of the membrane-bound enzyme cluster, and the most stable and active assembly has to be yet first membrane-triggered. PMID- 6284819 TI - Inhibition of ornithine decarboxylase and S-adenosylmethionine decarboxylase activities of S49 lymphoma cells by agents increasing cyclic AMP. AB - We have examined the regulation of two key enzymes that control polyamine biosynthesis-L-ornithine decarboxylase (ODC) and S-adenosylmethionine decarboxylase (SAMDC) - by agents increasing cAMP in S49 lymphoma cells. Incubation of wild type S49 cells with beta-adrenergic agonists (terbutaline or isoproterenol) inhibited ODC and SAMDC activities rapidly (less than 2 hr). more quickly than these agents arrested the cells in the G1 phase of the cell cycle. The beta-adrenergic antagonist propranolol blocked inhibition of ODC activity produced by isoproterenol, but only if added simultaneously or less than 4 hr after the agonist. Incubation of wild type S49 cells with cholera toxin or PGE1 also inhibited ODC activity. Decreases in ODC activity produced by beta adrenergic agonists, cholera toxin, PGE1 or dibutyryl cAMP were all enhanced by the phosphodiesterase inhibitor Ro 20-1724. Results of studies of ODC and SAMDC activity in S49 variants having lesions in the pathway of cAMP generation and action were as follows: kin- cells (which lack cAMP-dependent protein kinase activity) showed no inhibition of ODC by any agent; AC- cells (which have absent nucleotide coupling units in their adenylate cyclase system) only demonstrated inhibition in response to dibutyryl cAMP; UNC cells (which have deficient coupling of hormone receptors and adenylate cyclase) only demonstrated inhibition in response to dibutyryl cAMP and cholera toxin, and beta-depleted cells (which have a decreased number of beta-adrenergic receptors) responded as did wild type cells except for absent response to isoproterenol. We conclude that inhibition of ODC and SAMDC activity in S49 cells is an early response to agents that increase cAMP and that this action occurs via the "classical" pathways of activation of adenylate cyclase and protein kinase. These results in S49 cells contrast with evidence in other systems in which cAMP has been suggested to enhance polyamine biosynthesis, perhaps through alternative mechanisms. PMID- 6284820 TI - Distribution of cyclic nucleotides in layers of the cerebellum after decapitation. AB - The distribution of the cyclic nucleotides was examined in layers of the mouse cerebellum following decapitation. Cyclic AMP increased and cyclic GMP decreased in all three layers of the cerebellum examined. The increase in cyclic AMP in the granular layer was far greater than in either the molecular or white layers. In the cerebellum from control mice, the cyclic GMP concentration was highest in the molecular layer and lowest in the white layer. Even after decapitation, this cyclic GMP gradient in the cerebellum was maintained. PMID- 6284821 TI - Effect of varying dietary ratio of sodium and chloride on the responses of lactating dairy cows in hot weather. AB - Twenty-four lactating Holstein and Jersey cows were fed concentrate mixtures formulated to supplement total diets with 1) no added sodium chloride, 2) 1.0% sodium chloride, 3) 1.14% calcium chloride, and 4) 1.45% sodium bicarbonate during a 9-wk continuous trial, Experiment 1. Responses measured were body temperature, respiration rate, milk yield, milk composition, and 15 blood variables to evaluate acid-base status and express metabolic profile. Treatments affected body temperature and pH, bicarbonate, total carbon dioxide, and base excess of blood. No treatment affected milk yield and composition. Differences in dietary electrolytes had little influence on responses. In a second experiment immediately following the first, a 1-km walking stress was imposed during midafternoon on 3 days; treatment did not influence change in response variables or recovery from stress although Jerseys recovered faster than Holsteins. PMID- 6284822 TI - Lactating dairy cow responses to dietary sodium, chloride, and bicarbonate during hot weather. AB - Twelve Holstein and eight Jersey cows in the first half of lactation were assigned to one of four concentrate formulations to provide supplemental sodium salts in the total diet dry matter of 1) .5% sodium chloride, 2) .5% sodium chloride plus .72% sodium bicarbonate, 3) .5% sodium chloride plus .72% sodium bicarbonate, and 4) 1.44% sodium bicarbonate to measure effects on body temperature and respiration rate, milk yield and composition, and blood components that reflect acid-base balance. After data were adjusted for body weight, age, and a covariate based on differences within individuals in the standardization period, they were analyzed with a model that included breed, treatment, week, temperature-humidity index, and interactions. Cow fed sodium bicarbonate and no supplemental sodium chloride had lower body temperatures than the other groups. High sodium and chloride in basal diet and drinking water prevented the large change in dietary amount and ratio of these two electrolytes that treatments were designed to impose. More stringent control of dietary amounts and greater heat stress will be necessary to show effects of these elements on acid-base balance. PMID- 6284824 TI - Milker's nodules: differential diagnosis. PMID- 6284823 TI - Alteration of rumen fermentation, milk fat synthesis, and nutrient utilization with mineral salts in dairy cows. AB - The ability of mineral salts to alter rumen fermentation, rumen fluid dilution rate, milk fat synthesis, and nutrient utilization was investigated in dairy cows fed a high-concentrate, milk-fat depressing diet. Four rumen-fistulated Holstein cows were in a 4 X 4 Latin square design. Treatments consisted of: 1) basal (25% corn silage: 75% concentrate on a dry matter basis), 2) basal + 2.0% sodium chloride, 3) basal + 2.0% sodium bicarbonate, and 4) basal + 2.4% limestone. Addition of limestone to the basal diet reduced dry matter intake but increased efficiency of dietary nutrient utilization for milk synthesis. Sodium bicarbonate increased synthesis of milk fat. Sodium chloride also tended to increase milk fat synthesis whereas limestone had no effect. Milk yields (kg/day) and milk fat (%) for the four treatments were 1) 29.5, 2.40; 2) 29.3, 2.66; 3) 28.9, 3.26; and 4) 29.2, 2.32. Rumen fluid pH, dilution rate (%/hour), and molar percentage of acetate and propionate were: 1) 5.98, 10.3, 49 and 39; 2) 6.02, 12.4, 55 and 32; 3) 6.16, 12.2, 58 and 25; and 4) 5.92, 10.7, 51 and 38. Limestone was totally ineffective in altering ruminal pH, fluid dilution rate, molar percentages of acetate and propionate, and synthesis of milk fat. Improved feed efficiency for milk production after addition of limestone was related to an increase in starch digestion compared to the basal ration (95 versus 88%). PMID- 6284825 TI - Postirradiation malignant fibrous histiocytoma. A case report with a review of postirradiation sarcoma in the Japanese literature. PMID- 6284826 TI - [Changes in the cyclic adenosine monophosphate content of rat tissues in a neurodystrophic process]. PMID- 6284827 TI - Isolation, purification, and characterization of a complex heteroxylan from industrial wheat bran. PMID- 6284828 TI - Effect of lower body negative pressure on plasma ACTH and cortisol concentrations in man. PMID- 6284829 TI - Ectopic secretion of ACTH and met-enkephalin from a thymic carcinoid. AB - A 51-year old man presented with the classical features of Cushing's syndrome which had evolved over the previous 5 yr, and was found to have ACTH secretion from an atypical thymic carcinoid tumor. Tumor extract, assayed under conditions designed to prevent artefactual generation of peptides, was found to contain a wide variety of immunoreactive hormones including ACTH, alpha-MSH, CLIP, beta endorphin and met-enkephalin. The ACTH-related peptides were probably derived from a common precursor, pro-opiocortin, but the presence of met-enkephalin suggests the production of a separate type of precursor molecule. The tumor was locally invasive and, depsite subtotal excision and radiotherapy, continued to secrete large amounts of ACTH. Hypercortisolism was controlled longterm with pharmacological adrenal blockade and steroid replacement. PMID- 6284830 TI - Long term bromocriptine therapy in Cushing's disease. AB - Recently, bromocriptine has been successfully introduced as medical therapy for the pituitary hypersecreting syndromes of hyperprolactinemia and acromegaly. Subsequently, an inhibitory effect on ACTH secretion was reported using bromocriptine in Cushing's disease. While most studies have focused on the acute response to bromocriptine, few have investigated the response to chronic therapy with this agent. In this report, we describe a patient with Cushing's disease in whom an acute inhibition of ACTH and cortisol was demonstrated following bromocriptine and in whom the long term administration of bromocriptine as the sole therapy produced a transient beneficial response. Further pharmacologic testing suggested this inhibitory effect on the hypothalamic-pituitary-adrenal axis was mediated through dopaminergic stimulation. PMID- 6284831 TI - Familial hypocalciuric hypercalcemia. Report of a new family. AB - We report a new family with familial hypocalciuric hypercalcemia (FHH) composed by 55 living members. Of 38 studied, 10 have been found to be affected by FHH. Differences between FHH and primary hyperparathyroidism are emphasized; lack of clinical features, relative hypocalciuria for the concomitant hypercalcemia and low phosphate excretion index. The PTH and urinary cAMP are normal. It is noteworthy that the disease is benign. None of our patients have undergone surgery, and all of them are asymptomatic. PMID- 6284832 TI - Oxidative metabolism and bactericidal capacity of polymorphonuclear leukocytes from normal young and aged adults. AB - The ability of polymorphonuclear leukocytes (PMNs) from young and old individuals to reduce nitroblue tetrazolium, to kill Staphylococcus aureus in vitro, and to generate superoxide was determined. Investigation of PMNs from 202 humans, aged 26 to 95 years, failed to demonstrate an age relationship in their ability to reduce nitroblue tetrazolium. The ability of PMNs from 20 young (M age 34 years) individuals to kill S. aureus in vitro did not differ from cells from 18 elderly (M age 68 years) persons. Mean production of superoxide in response to stimulation with latex particles was, however, significantly lower (P less than .001) in cells from elderly adults. These results suggest a heterogeneity in any age-associated defect in PMN function. Several aspects of PMN function need to be evaluated in order to describe overall PMN function in the elderly. PMID- 6284833 TI - Gastric xanthoma: histologic similarity to signet ring cell carcinoma. AB - Gastric xanthomas (GX) are pale yellow nodules or plaques, usually less than 3 mm in size, in the gastric mucosa. They are frequently found in groups, most often along the lesser curvature and pyloric regions. They consist of liquid-laden histiocytes in the lamina propria occasionally extending into the submucosa. There is no correlation between GX and hypercholesterolemia. The lesion is of little significance, except as it may be misdiagnosed as carcinoma. We present two cases of GX, one initially thought histologically to be a signet ring cell carcinoma. With the paucity of reports of GX in the medical literature and the potential for incorrect diagnosis, we emphasize the importance of recognition and accurate histologic diagnosis of GX. PMID- 6284834 TI - Linkage of two enzyme loci in fishes of the genus Xiphophorus (Poeciliidae). Guanylate kinase-2 (GUK2) and glyceraldehyde-3-phosphate dehydrogenase-1 (GAPD1) designated as linkage group III. AB - Electrophoretic variation ascribable to two enzyme loci, coding for a guanylate kinase (GUK2) and a glyceraldehyde-3-phosphate dehydrogenase (GAPD1), was observed in three species of fishes of the genus Xiphophorus. Electrophoretic patterns in F1 hybrid heterozygotes suggested a monomeric subunit structure for GUK2 and confirmed a tetrameric structure for GAPD1. Variant alleles at the two loci exhibited normal Mendelian segregation in backcross hybrids. Linkage analyses indicate estimated recombination of GUK2-7.6 percent-GAPD 1. This group (designated linkage group III) was shown to assort independently from the 7 loci comprising linkage groups I and II and from 26 other informative markers, within the limits of the data. Difficulties inherent in establishing homology with linkage groups in other species in cases involving presumed gene duplication are discussed. PMID- 6284835 TI - Salmonella organisms in garden fertilizers of animal origin. AB - Of 120 specimens of garden fertilizers of animal origin purchased in retail shops, 40 (33.3%) were found to be contaminated with salmonella organisms. Untreated bone meal (53.1%) was the most heavily contaminated but 25% of specimens of this product classed as heat-treated or sterilized were positive. In all, 32 serotypes were identified. PMID- 6284836 TI - Guanidine and heat sensitivity of foot-and-mouth disease virus (FMDV) strains. AB - A study of the ability of 49 strains of FMD virus to replicate in BHK-21 monolayer cells maintained under a standard agar overlay containing 5.2 mM guanidine hydrochloride and to withstand heat inactivation at 54 degrees C for 1 h showed that strains belonging to serotypes C, O and Asia 1 were generally more resistant to guanidine and heat stable than the SAT 1, 2 and 3 serotypes. The type A viruses as a whole occupied an intermediate position between these two groups. In vitro passage in BHK-21 cells influenced the guanidine sensitivity of 3(O, C and SAT 3) of the 7 FMD serotypes suggesting that this is not a stable genetic marker. Heat stability of the FMD viruses, however, did not change on passage, suggesting that this is a stable characteristic inherent in any homogeneous FMD virus population. PMID- 6284837 TI - Occurrence of viruses in human stools in the Ahaggar (Alberia). AB - From October 1977 to May 1980, 243 stools collected in sedentary and semi-nomadic populations of the Ahaggar (Algerian Sahara) were examined using immunoelectronmicroscopy and tissue culture inoculation. Immunoelectronmicroscopy revealed the presence of rotaviruses in 8, coronaviruses in 26, adenoviruses in 5 and small round viruses in 4. Enteroviruses were isolated in tissue culture from 24 stools. Rotaviruses were present in the Ahaggar but were associated with little acute enteric disease. The high frequency of coronaviruses both in gastroenteritis patients and in patients without disease was surprising. The prevalence of enteroviruses in this hyperarid zone was similar to or higher than that found in noticeably more human countries. Further systematic bacterial, viral and parasitic examinations are required to clarify the role of the above viruses in the aetiology of gastroenteritis in this region. PMID- 6284838 TI - A comparison of the levels of faecal indicator bacteria in water and human faeces in a rural area of a tropical developing country (Sierra Leone). AB - The levels of faecal coliforms (FC), indole-positive FC (presumptive Escherichia coli), faecal streptococci (FS), Streptococcus faecalis and Clostridium perfringens in the natural water sources used by 29 rural settlements in Sierra Leone were investigated. Levels of the same indicators in human faeces were also investigated. The incidence of Salmonella spp. in both habitats and the temperature, pH and conductivity of water sources were also recorded. All water sources were contaminated with the indicator bacteria, mean numbers of which occurred in the relationship FC greater than presumptive E. coli congruent to FS congruent to C. perfringens greater than S. faecalis. FC were also predominant in human faeces, the relationship of means being FC congruent to presumptive E. coli greater than FS greater S. faecalis greater than C. perfringens. The need for confirmation of FC counts obtained from water sources was indicated by the large number of positive tubes produced in the FC multiple-tube dilution test from some samples which could not be confirmed as presumptive E. coli. Salmonella spp. Were isolated from 13 water sources and 6% of faecal samples. Mean water temperature was high (26.2 degrees C), pH low (5.04) and conductivity low (34 microS cm-1). Presumptive E. coli was considered the most appropriate indicator of faecal pollution of the types of water investigated. PMID- 6284839 TI - [The results of combined treatment for testicular tumours. Report of 12 cases (author's transl)]. AB - Over a ten year period, the authors have collected 120 cases of germinal tumours of the testis (35 seminomas and 85 non-seminomatous tumours). The largest single group was of mixed non-seminomatous tumours. The treatment of pure seminomas gave 100% good results by a combination of unilateral orchidectomy, radiotherapy and, in the case of stage III tumours with spread, chemotherapy. The prognosis of non seminomatous tumours was dependent essentially upon the stage of spread. During the past three years, the best results of chemotherapy have been obtained with the combination of vincristin, bleomycin and platinum (1978, 1979, 1980). In his time, at stage I and stage II, all the patients are alive. Treatment consisted of a minimum of unilateral orchidectomy and retroperitoneal lymphadenectomy, Chemotherapy was envisaged only for those tumours with evidence of lymph node involvement. At stage III, they lost 3 patients out of 7, this being an excellent result bearing in mind the gravity of such tumours. PMID- 6284840 TI - On the mechanism of enhanced monocyte and neutrophil cytotoxicity in severe psoriasis. AB - Monocyte and neutrophil function assessed as antibody-dependent cell-mediated cytotoxicity (ADCC) using IgG-sensitizing human erythrocytes as target cells was enhanced in patients with severe psoriasis when compared to healthy controls. We found significant correlation between increased monocyte ADCC and increased neutrophil ADCC, No differences in basal cAMP levels and cAMP responses during initiation of the ADCC reaction was observed between psoriatics and normals. Also degranulation determined as lysozyme release during ADCC was normal. In contrast, the increase in ADCC was significantly correlated to an enhanced hexose monophosphate shunt activation in the effector cells during the cytotoxic reaction. Activity of enzymes responsible for the respiratory burst was not altered in psoriasis since superoxide production after stimulation with phorbol myristate acetate was normal. Likewise, oxygen consumption and degranulation following phagocytosis of opsonized zymosan particles in neutrophils was found normal in psoriasis. Since monocytes showed increased binding of IgG-sensitized erythrocytes these data indicate that the enhanced monocyte and neutrophil ADCC is caused by an enhancement of the respiratory burst possibly induced by increased Fc receptor activity. PMID- 6284841 TI - Kaposi's sarcoma: absence of cytomegalovirus antigens. AB - Using an immunohistochemical method, we could not detect cytomegalovirus antigens in the tissue of Kaposi's sarcoma but could in the cells of control sections from tissues known to be infected by that virus. Our results are consistent with the hypothesis that high antibody titers to cytomegalovirus found in patients with Kaposi's sarcoma are not due to infection of neoplastic cells by the virus, as a secondary event; our data supports the conclusions of others that exposure to cytomegalovirus may be a primary event in the pathogenesis of Kaposi's sarcoma. PMID- 6284842 TI - [Transcription and RNA processing of SV40 DNA (author's transl)]. PMID- 6284843 TI - [Genome structure of poliovirus (author's transl)]. PMID- 6284844 TI - [Surface ultrastructural study on the human endometrial interstitium using digestion method (author's transl)]. AB - A scanning electron microscopic study on the human endometrium, using Evan's method and modifications, revealed the following findings. a) 8N . HCl Collagenase method: High-speed digestion of the specimen made the determination of the adequate digestion time difficult, due to conc. HCl and high processing temperature. b) 1N . HCl-Collagenase method: Long digestion time produced many artifacts as shrinkage of the cells or formation of small holes on them. c) Tripsin-Collagenase method: Tissue breaking often resulted before the adequate digestion. d) Hyaluronidase-Collagenase method: This is most satisfactory in classifying glands and vessels, and digestion time is the shortest of all methods. PMID- 6284845 TI - [Specific radioimmunoassay and physiological significance of unconjugated 2 hydroxyestrone and 2-hydroxyestradiol-17 beta in human blood (author's transl)]. AB - Recently it has been attached importance to the physiological function of catecholestrogens. To elucidate it a specific radioimmunoassay of conjugated 2 hydroxyestrone (2-OHE1) and 2-hydroxyestradiol-17 beta (2-OHE2) in human plasma was attempted. After extracting with ethyl acetate, samples were purified by a short Sephadex LH-20 columnchromatography and determined using the antiserum to 2 OHE1-17(O-carboxymethyl) oxime-bovine serum albumin conjugate. The following results were obtained: 1) A long Sephadex LH-20 columnchromatography which was used for the purification of catecholestrogens produced chemically, had a superior faculty in separation and high capacity 2) The antiserum cross-reacted 26.4% with 1-OHE2, but less than 1% with other steroid hormones. 3) The sensitivity of the method was around 10pg in both 2-OHE1 and 2-OHE2 assays. The method blank determined using 2ml plasma of bilaterally adreno-oophrectomized women was below 15 pg/ml in both 2-OHE1 and 2-OHE2 assays (n = 24). 4) The coefficient of variation in both accuracy and between-assay precision of the method was less than 17%. 5) The plasma 2-OHE1 concentration was below 15 pg/ml in normal men (n = 8) and non-pregnant women (n = 13). The concentration in pregnant women was 20 +/- 8 pg/ml (SD, n = 7) in 1st trimester of pregnancy, 58 +/- 13 (n = 3) in 2nd trimester and 177 +/- 66 (n = 12) in 3rd trimester. The E1/2-OHE1 and E2/2-OHE1 ratios in 3rd trimester of 9 pregnant women were 19.7 +/- 11.9(SD) and 78.0 +/- 27.8, respectively. 6) The plasma 2-OHE2 concentration was below 15 pg/ml in normal men (n = 8), non-pregnant women (n = 8), and 1st to 2nd trimester (n = 10) and 15 +/- 9 pg/ml (SD, n = 13) in 3rd trimester of pregnant women. PMID- 6284846 TI - Vulvar carcinoma. A case of DDP-induced neurotoxicity in chemotherapy. PMID- 6284847 TI - Renin and aldosterone responses to short-term NaCl or NaHCO3 loading in man. AB - We compared the effects of 500 ml of 0.15M NaCl or 750 ml of 0.15M NaHCO2, infusions on PRA and PAC in 16 normal NaCl-restricted men. More positive sodium balance, greater natriuresis, and lower serum chloride, potassium, and hydrogen ion concentrations were observed after NaHCO3 infusion. PRA decreased (13.8 +/- 1.0 ng/ml angiotensin I per hour to 6.6 +/- 0.70) and PAC did not change (98 +/- 15 ng/dl to 86 +/- 15) after NaCl infusion. Conversely, PRA did not change (9.9 +/- 1.6 to 10.1 +/- 1.6) and PAC decreased (85 +/- 9 to 44 +/- 5) after NaHCO3 infusion. Plasma cortisol declined in both groups. These results suggest that in sodium-restricted man chloride is more important than sodium for the suppression of PRA by NaCl loading and that potassium or hydrogen ion is a more sensitive modulator of aldosterone secretion than is renin. PMID- 6284848 TI - Granulocyte phagocytosis and killing virulent and avirulent serotypes of Streptococcus pneumoniae. AB - Five commonly isolated Streptococcus pneumoniae serotypes (3, 6, 14, 19, and 23) and five rarely found serotypes (31, 35, 36, 42, and 43) were compared to elucidate whether increased resistance against granulocyte phagocytosis and killing could explain the restricted number of pneumococcal serotypes found in infections. There was a great variation in sensitivity among the serotypes to granulocyte killing. No consistent pattern was found when pathogenicity and resistance to granulocytes were compared. The results do not indicate that the increased tendency of pathogenic pneumococcal serotypes to cause infections is due to increased resistance to granulocytes. Monocyte killing of some pneumococal serotypes (6, 19, 23, 35, and 43) was also studied and found very similar to granulocyte killing. Defective granulocyte kiling of encapsulated pneumococci was due to impaired phagocytosis. Moreover, no correlation was found between the sensitivity of the serotypes to isolated intragranulocytic microbial systems (i.e., MPO, hydrogen peroxide, or CCP) and the sensitivity to killing by intact granulocytes or pathogenicity. The significance of both the classical and alternative complement pathways for pneumococcal opsonization was indicated by reduced, the residual phagocytosis in C2-deficient and MgEGTA-chelated serum. PMID- 6284849 TI - Unilateral odontodysplasia with ipsilateral hypoplasia of the mid-face. A case report. AB - A ten-year-and-1-month-old patient with regional odontodysplasia in the right maxilla is presented. The tooth malformation extended as far as the midline. The teeth 654321 + were markedly affected and exhibited a small indistinctly mineralized border with the 7 + being somewhat hypermineralized. The affected teeth in the respective quadrant had incomplete, retarded root formation. In addition, the patient had an obvious hypoplasia of the zygomatico-maxillary complex on the right. Ischaemia due to insufficient blood supply by the maxillary artery on the right is considered to be a possible cause of the disturbance described, and this is discussed. PMID- 6284850 TI - Effect of Ca2+ deprivation on release of luteinizing hormone induced by luteinizing hormone releasing hormone from female rat pituitary glands in vitro. PMID- 6284851 TI - Cytoplasmic and nuclear progesterone receptors in mouse mammary tumours during pregnancy determined by an improved hydroxylapatite assay. PMID- 6284852 TI - Comparative pharmacology of oestrogens and catechol oestrogens: actions on the immature rat uterus in vivo and in vitro. AB - The effects of primary and catechol oestrogens on the uterus of the immature rat were compared. Because differences between the in-vivo and in-vitro oestrogenic actions of catechol oestrogens on the secretion of LH had been observed, their effects on a peripheral target organ, the uterus, were examined under similar conditions. In-vivo effects were assessed by measurement of uterine weight, induction of uterine cytoplasmic progestogen receptors, and by histological examination. In-vitro actions were determined by measurement of oestrogen specific induced protein. It was found that the uterotrophic effects in vivo of 4 hydroxyoestradiol were indistinguishable from those of oestradiol whereas 2 hydroxyoestradiol was only weakly oestrogenic and 2-hydroxyoestrone had no effect. However, in vitro, 2-hydroxyoestradiol was as effective as 4 hydroxyoestradiol or oestradiol in stimulating synthesis of uterine induced protein, and 2-hydroxyoestrone, although less potent than oestradiol, had a significant effect. These results were consistent with the observed effects on the secretion of LH. The differences between in-vivo and in-vitro uterotrophic properties of catechol oestrogens can be explained on the basis of known pharmacokinetic factors. PMID- 6284853 TI - An assay of the capacity of biological fluids to stimulate renal glucose-6 phosphate dehydrogenase activity in vitro as a marker of their ability to inhibit sodium potassium-dependent adenosine triphosphatase activity. PMID- 6284854 TI - Genetic interactions in the spontaneous production of endogenous murine leukemia virus in low leukemic mouse strains. AB - The spontaneous expression of ecotropic murine leukemia virus (MuLV) in spleen cells of BALB/c, C57BL/6 (B6), and derivative mice was examined as a function of age. The patterns of spontaneous virus induction in vivo correlate with the patterns of virus induction in vitro, which result from the action of two loci, Inc-l and Inb-l (7). Whereas mice carrying Inc-l or Inb-l have similar phenotypes in vitro, they have significantly different phenotypes in vivo. Mice of the Inb l+/+ genotype, e.g., B6, rarely expressed MuLV, and the titer of MuLV recovered from rare MuLV-positive mice of this genotype was usually low. Mice of the Inc l+/+ genotype, e.g., BALB/c, expressed low amounts of MuLV early in life, however, from 6-12 mo of age approximately one-half of the Inc-l+/+ mice expressed virus, frequently of high titer. Equal numbers of N-tropic and B-tropic MuLV were recovered from Inb-l+ mice, but predominantly N-tropic MuLV was recovered from Inc-l+ mice. Strains that carry dominant (+) alleles at both Inc-l and Inb-l show higher titers of MuLV earlier in life than strains that carry only Inc-l or Inb-l. The presence of dominant alleles at both loci results in the appearance of predominantly N-tropic virus early in life. These results demonstrate that the principal determinants of spontaneous virus expression in these low leukemic strains of mice are the In loci or genes linked to them. A further inference that can be drawn from these studies is that the appearance of B-tropic virus is by no means a random process but rather results from predictable patterns of MuLV expression and alteration. PMID- 6284855 TI - Transmission and scanning electron microscopic studies of the human sperm chromatin decondensed by micrococcal nuclease and salt. AB - The human sperm chromatin was gently decondensed by treating the sperm head sequentially with micrococcal nuclease and 2 M NaCl. All histones, about 10% of DNA, and a small amount of degraded protamines were released into the soluble fraction, leaving mainly nucleoprotamines in the pellet fraction. Transmission and scanning electron microscopic studies revealed that the nucleoprotamine pellet consisted of chromatin cords of two dimensions, viz., 330- to 420-A and 650- to 1200-A thick cords laced together by very fine strands of 60- to 80-A fibers; both types of cord appeared knobby and had zig-zag patterns throughout their length. It appeared that these cords were derived from two types of sperm heads of approximately equal population; one type contained chiefly the thick cords and the other chiefly the thin cords. Further treatment of the pellet nuclease-NaCl with urea and mercaptoethanol resulted in the dissociation of the thick into the thin cords and unravelling of the thin cords into smaller sized fibers; whereas the treatment of the pellet nuclease-NaCl with DNAase I resulted in the disappearance of the 60- to 80-A fibers, and the remaining cords were chiefly of thick type together with the sperm head exoskeletons. From these results the packing order of the chromatin in human sperm was proposed. PMID- 6284857 TI - Leveen peritoneal jugular shunt in patients with ascites. PMID- 6284856 TI - Amphotericin B and the elasmobranch rectal gland: implications for the relationship between oxygen consumption and ion transport. AB - In slices of the dogfish rectal gland, amphotericin B produced increases in oxygen consumption and ouabain binding similar to those produced by cyclic AMP, except that the increases induced by the latter were inhibited by furosemide whereas those resulting from the addition of amphotericin B were unaffected by the diuretic. Unlike cyclic AMP, however, amphotericin B failed to stimulate secretion by the isolated perfused gland. These results support the suggestion that the increases in ouabain binding and oxygen consumption produced by cyclic AMP result simply from an increased sodium entry into the cells, and as such can be mimicked by amphotericin B, whereas secretion rate itself depends on some additional, cyclic AMP-induced process. The implications of this for determinations of the relationship between rates of oxygen consumption and ion transport are discussed. PMID- 6284858 TI - Evidence for a population of sleepy sodium channels in squid axon at low temperature. AB - We have studied the effects of temperature changes on Na currents in squid giant axons. Decreases in temperature in the 15-1 degrees C range decrease peak Na current with a Q10 of 2.2. Steady state currents, which are tetrodotoxin sensitive and have the same reversal potential as peak currents, are almost unaffected by temperature changes. After removal of inactivation by pronase treatment, steady state current amplitude has a Q10 of 2.3. Na currents generated at large positive voltages sometimes exhibit a biphasic activation pattern. The first phase activates rapidly and partially inactivates and is followed by a secondary slow current increase that lasts several milliseconds. Peak Na current amplitude can be increased by delivering large positive prepulses, an effect that is more pronounced at low temperatures. The slow activation phase is eliminated after a positive prepulse. The results are consistent with the hypothesis that there are two forms of the Na channel: (a) rapidly activating channels that completely inactivate, and (b) slowly activating "sleepy" channels that inactivate slowly if at all. Some fast channels are assumed to be converted to sleepy channels by cooling, possibly because of a phase transition in the membrane. The existence of sleepy channels complicates the determination of the Q10 of gating parameters and single-channel conductance. PMID- 6284859 TI - Induction of dark-adaptive retinomotor movement (cell elongation) in teleost retinal cones by cyclic adenosine 3','5-monophosphate. AB - In the teleost retina, the photoreceptors and retinal pigment epithelium (RPE) undergo extensive movements (called retinomotor movements) in response to changes in light conditions and to an endogenous circadian rhythm. Photoreceptor movements serve to reposition the light-receptive outer segments and are effected by changes in inner segment length. Melanin granule movements within the RPE cells provide a movable melanin screen for rod outer segments. In the dark (night), cones elongate, rods contract, and pigment granules aggregate to the base of the RPE cell; in the light (day), these movements are reversed. We report here that treatments that elevate cytoplasmic cyclic adenosine 3',5' monophosphate (cAMP) provoke retinomotor movements characteristic of nighttime dark adaptation, even in bright light at midday. To illustrate this response, we present a quantitative description of the effects of cyclic nucleotides on cone length in the green sunfish, Lepomis cyanellus. Cone elongation is induced when light-adapted retinas are exposed to exogenous cAMP analogues accompanied by phosphodiesterase (PDE) inhibitors (either by intraocular injection or in retinal organ culture). Cone movements is not affected by cyclic GMP analogies. Dose response studies indicate that the extent, but not the rate, of cone elongation is proportional to the concentration of exogenous cAMP and analogue presented. As has been reported for other species, we find that levels of cAMP are significantly higher in dark- than in light-adapted green sunfish retinas. On the basis of these observations, we suggest that cAMP plays a role in the light and circadian regulation of teleost cone length. PMID- 6284861 TI - Tonic contraction and the control of relaxation in a chemically skinned molluscan smooth muscle. AB - The same functional states that characterize the living anterior byssus retractor muscle (ABRM) from Mytilus edulis can be initiated in the saponin-treated (chemically skinned) muscle preparation under controlled biochemical conditions. A tonic contraction was induced if the concentration of free Ca2+ was above approximately 10(7) M in the presence of Mg2+ and ATP. Maximum tension development was achieved at a Ca2+ concentration of approximately 10(4) M. Within these Ca2+ concentrations tension was always associated with the presence of 'active state," as indicated by a high recovery of tension after a quick release in muscle length. Tonic tension, and the associated active state was maintained for hours during these conditions irrespective of variations in both ionic strength and pH. Reduction of the Ca2+ concentration to below threshold for tension initiation during a tonic contraction immediately switched off the active state and relaxation of the muscle preparation resulted. However, the rate of relaxation was extremely low, leaving a substantial fraction of tension in the absence of active state. Both 5-hydroxytryptamine (5-HT) and cAMP accelerated this slow relaxation in the absence of Ca2+. Thus, this state was considered equivalent to the 'catch state" in the living ABRM. In the presence of Ca2+ concentrations above 10(7) M, cAMP did not affect either the maximum tension developed or the Ca2+ sensitivity of the chemically skinned muscle preparation. PMID- 6284860 TI - Effects of cyclic adenosine 3',5'-monophosphate on photoreceptor disc shedding and retinomotor movement. Inhibition of rod shedding and stimulation of cone elongation. AB - As a test of the hypothesis that cyclic nucleotides play a role in the regulation of retinomotor movements and disc shedding in the photoreceptor-pigment epithelial complex, we have used an in vitro eyecup preparation that sustains both disc shedding and cone retinomotor movements, Eyecups were prepared in white light from animals in which both shedding and cone movement had been blocked by 4 d of constant-light treatment. In eyecups incubated for 3 h in light, disc shedding was negligible and cones remained in the light-adapted (contracted) position. In eyecups incubated in darkness, however, a massive shedding response (dominated by rod photoreceptors) was induced, and at the same time cone photoreceptors elongated to their dark-adapted position. In eyecups incubated in light dbcAMP promoted cone elongation and thus mimicked darkness; the dbcAMP effect was potentiated by the phosphodiesterase inhibitors papaverine and 3 isobutylmethylxanthine. In eyecups incubated in darkness, on the other hand, both phosphodiesterase inhibitors and dbcAMP reduced the phagosome content of the pigment epithelium. The effects of dbcAMP on the cone elongation and rod shedding appear to be specific in that dbcGMP, adenosine, and adenosine 5'-monophosphate had no significant effect. Our results suggest that cAMP plays a role in the regulation of both retinomotor movements and disc shedding. PMID- 6284862 TI - Bis-quaternary ammonium blockers as structural probes of the sarcoplasmic reticulum K+ channel. AB - A series of n-alkyl-bis-alpha,omega-trimethylammonium (bisQn) compounds was synthesized, and their ability to block K+ currents through a K+ channel from sarcoplasmic reticulum was studied. K+ channels were inserted into planar phospholipid membranes, and single-channel K+ currents were measured in the presence of the blocking cations. These bisQn compounds block K+ currents only from the side of the membrane opposite to the addition of SR vesicles (the trans side). The block is dependent on transmembrane voltage, and the effective valence of the block (a measure of this voltage dependence) varies with the methylene chain length. For short chains (bisQ2-bisQ5), the effective valence decreases with chain length from 1.1 to 0.65; it then remains constant at approximately 0.65 for bisQ5 to bisQ8; the effective valence abruptly increases to 1.2-1.3 for chains of nine carbons and longer. For the compounds of nine carbons and longer, the discrete nature of the block can be observed directly as 'flickering noise" on the open channel. The kinetics of the block were studied for these long-chain blockers. Both blocking and unblocking rates of the blockers vary with chain length, with the blocking rate showing the strongest variation--an increase of 2.8-fold per added methylene group. All of the voltage dependence of the binding equilibrium resides in the blocking rate, and none in the unblocking rate. The results imply that 65% of the voltage drop within the channel occurs over a distance of 6-7A, and that the short-chain blockers bind in a bent-over conformation with both charges deeply inside the channel. PMID- 6284864 TI - Effect of a mutation in the VSV G protein on the synthesis and maturation of nucleocapsids. AB - The tsO45 (V) mutant of vesicular stomatitis virus (VSV) has been used to demonstrate that discrimination between positive (+) and negative (-) strand nucleocapsids in the process of budding mature virions is maintained despite the lack of detectable G protein in either the plasma membrane or the virus membrane. These data indicate that the VSV G protein plays no active role in the discrimination between negative- and positive-strand nucleocapsids which will mature to virions. The synthesis of intracellular nucleocapsids in tsO45 (V) infected cells at permissive and non-permissive conditions was also examined in comparison with that of the wild-type virus. No differences were observed in intracellular nucleocapsid quantity or polarity in any of these cases despite a 97% inhibition of virus yield in the case of tsO45 (V) at non-permissive temperature. PMID- 6284863 TI - The transition from HK to LK phenotype in the red cells of newborn genetically LK lambs. AB - Red cells from newborn lambs were separated into different age populations by centrifugation, and cells with fetal hemoglobin (Hb) were distinguished from those with adult Hb by an acid elution technique. Changes were followed during development in rates of K+ transport (active and passive), numbers of Na+/K+ pump sites per cell, cell volumes, and numbers of Lp and L1 antigen sites per cell. These changes were correlated with the percentage of cells with adult hemoglobin. (The Lp and L1 antigens are associated with K+ transport in that specific alloantibody against Lp, anti-Lp, stimulates active transport, and anti-L1 inhibits passive transport.) Active K+ transport decreased during development because of a decline in number of Na+/K+ pumps (from measurements of ouabain binding) and because of an alteration in the affinity of the pumps for intracellular K+ (from kinetic studies in which the intracellular K+ concentration was varied). Cells with fetal Hb had fewer Lp sites and were larger than cells with adult Hb. As transport properties changed, the number of Lp sites increased and continued to increase after all the cells had adult Hb Cells with fetal Hb had as many L1 sites as lamb cells with adult Hb, but the number of L1 sites was less than those found previously for adult sheep. A population of small cells with intermediate K+ concentration and intermediate numbers of Lp sites appeared soon after birth. The various points of evidence suggested that the developmental process leading to cells with adult transport properties was a gradual one and did not coincide precisely with the switch from fetal to adult Hb. PMID- 6284865 TI - Electrophoretic analysis of RNA segments of human rotaviruses cultivated in cell culture. AB - Human rotaviruses (HRVs) derived from stools were cultivated to high titres in an established cell line (MA-104) using the rotary culture system. Analysis of the 11 double-stranded RNA segments of the culture-adapted HRVs was carried out by polyacrylamide gel electrophoresis. Tissue culture-adapted HRVs, uncloned or cloned, had the same RNA gel patterns as those of the original HRVs from the individual stool specimens. The migrations of the RNA segments from the culture adapted HRVs were markedly different from that of the RNA segments from calf rotavirus (Lincoln strain). Considerable heterogeneity in electrophoretic migration of the RNA was found among eight strains of the HRVs grown in cell culture. These results confirmed that viruses isolated from stool specimens were indeed of human origin and were not the result of laboratory contamination with tissue culture-adapted calf rotavirus or other viruses. PMID- 6284866 TI - Avian nephroblastoma virus MAV-2(N) and avian osteopetrosis virus MAV-2(O) are genetically distinct. AB - Avian myeloblastosis virus consists of a mixture of a defective leukaemia virus and several non-defective associated avian leukosis viruses. The genomes of two of the associated avian leukosis viruses were examined in this study and were chosen because one of them, MAV-2(N), induces predominantly nephroblastoma, while the other, MAV-2(O), induces predominantly osteopetrosis. Competitive hybridization studies employing labelled virion RNA and DNA from normal and malignant tissue failed to demonstrate a difference the genomes. However, examination of ribonuclease T1-resistant oligonucleotide maps revealed that MAV 2(N) RNA had five oligonucleotide fragments which were not present in the MAV 2(O) genome. Poly(A) selection of the oligonucleotides at the 3' end of the genome showed that the fragments unique to MAV-2(N) were not present at this end of the genome. These results suggest that two viruses differing in oncogenic manifestation also differ in genome composition. PMID- 6284867 TI - Time of synthesis and cellular location of Epstein-Barr virus-specified polypeptides radioimmunoprecipitated from iododeoxyuridine-induced Burkitt lymphoma/somatic cell hybrids. AB - The Burkitt lymphoma/somatic cell hybrid D98/HR-1 contains repressed Epstein-Barr virus (EBV) genomes which can be induced by 5-iodo-2'-deoxyuridine (IdUrd) to express the EBV early antigen (EA) and the virus capsid antigen (VCA). Nuclear and cytoplasmic fractions prepared from uninduced, partially induced and fully induced hybrid cells were radioimmunoprecipitated with human sera having high antibody titres against EA and VCA. Three polypeptides, with mol. wt. 127 x 10(3), 115 x 10(3) and 84 x 10(3) (127K, 115K and 84K), were detected in cells induced to express EA. All three polypeptides continued to be synthesized late in infection. One of the polypeptides was predominantly nuclear, one predominantly cytoplasmic, and one found in both fractions. Three polypeptides, with mol. wt. 144K, 137K and 50K, were detectable only in cells induced to express both EA and VCA. All three polypeptides were found predominantly in the nuclear fraction. PMID- 6284868 TI - Synthesis of message and genome RNAs in vitro by Sendai virus-infected cell nucleocapsids. AB - Purified Sendai virus nucleocapsids isolated from infected cells were used to programme a transcription system in vitro to study virus-specific RNA synthesis. The RNA products were analysed for size by centrifugation before and after denaturation with formamide or glyoxal. The polarity of the products [message (+) or genome (-) strands] was analysed by RNA-RNA hybridization. The non-denatured RNA products sedimented in three groups: 7S to 22S single-stranded RNA transcripts and two partially ribonuclease-resistant complexes. One complex, representing 12% of the total product, sedimented at 26S to 36S. After denaturing the 26S to 36S complex to single-stranded molecules, about half of the RNAs sedimented at 25S to 54S and about half at 6S to 24S. The second complex, representing about 13% of the total RNA product, sedimented at 42S to 52S. After denaturing, about 10% of the single-stranded RNAs sedimented at 38S to 52S and about 90% sedimented at 6S to 19S. In hybridization studies, single-stranded RNAs that sedimented at less than 19S were predominantly of message sense (+ strand), whereas RNAs that sedimented at 25S to 54S were a mixture of genome and anti genome type. These results show that transcription and replication activities in vitro were associated with Sendai virus nucleocapsids obtained from infected cells and that some of the reaction products approached genome size. PMID- 6284869 TI - Analysis of viral and defective-interfering nucleocapsids in acute and persistent infection by rhabdoviruses. AB - We have isolated and characterized the RNA of intracellular virus nucleocapsids recovered from a number of cell cultures persistently infected with rabies virus or vesicular stomatitis virus (VSV). VSV persistent infections in BHK21, L cells and Aedes albopictus (mosquito) cells generally showed the presence of large amounts of defective-interfering (DI) nucleocapsid RNA and much smaller amounts of standard (B) nucleocapsid RNA. Persistent infections of BHK21 cells by two rabies virus strains, challenge virus standard (CVS-11) or HEP-Flury, were followed for several months during which time the ratio of DI to B nucleocapsid RNA cycled dramatically. We also observed coordinated fluctuations in the absolute amount of incorporation of [3H]uridine into virus nucleocapsid RNA. Total incorporation was generally highest following a decrease in the relative amount of DI nucleocapsid RNA synthesis. At no time were DI nucleocapsids absent in any of the persistently infected cultures. PMID- 6284870 TI - Antibodies to gastroenteritis viruses in cystic fibrosis patients. AB - Infectivity of certain enteric viruses including rotavirus is profoundly affected by proteolytic enzymes. To test whether cystic fibrosis patients, possessing chronically decreased levels of pancreatic enzymes, show altered susceptibility to gastroenteritis viruses, we examined sera from patients and controls for antibodies to two major pathogens. In cystic fibrosis patients, normal rotavirus antibody levels were found and Norwalk virus antibody prevalence was unchanged. PMID- 6284871 TI - Hepatitis-B virus infection in black children with hepatocellular carcinoma. AB - The hepatitis B virus (HBV) status of six unselected South African Black children, aged 10 to 16 years, with hepatocellular carcinoma (HCC) was investigated. The characteristics of the tumor were similar to those seen in Black adults. Hepatitis B surface antigen and antibody to the core antigen were present in the serum of all the children, and hepatitis Be antigen was detected in four of the children. The serum of the two patients without e antigen was positive for e antibody, and all sera were negative for hepatitis B surface antibody. Taken in conjunction with the HBV status of southern African Black adults with HCC, these findings raise the possibility that nearly all, if not all, Black patients with this tumor are or have been infected with HBV. Moreover, if HBV is oncogenic, tumor formation may occur within 10 years of the initial infection, which occurs in early childhood in southern African Blacks. PMID- 6284872 TI - Complement components in kidney allograft recipients: relationship to cytomegalovirus infection. AB - Serum complement profiles of 19 renal transplant recipients were studied serially before and after renal transplantation to determine if there is any relationship between cytomegalovirus (CMV) infection and alteration in the serum complement (C) levels. Most patients had low serum C3 and C4 levels before transplantation, but afterwards these levels increased in some patients. Clq was elevated before and after transplantation. Factor B was low before transplantation, but after surgery, its level approached normal in patients without cytomegalovirus viremia, whereas in those with viremia, Factor B level became even more depressed. Circulating Factor B fragments were found in the sera of five out of seven patients with active cytomegalovirus infection, which suggests activation of the alternative pathway possibly related to active CMV infection. PMID- 6284873 TI - Lithium chloride-induced taste aversion in C57BL/6J and DBA/2J mice. AB - Male C57BL/6J and DBA/2J mice (Mus musculus) were tested in a one-bottle conditioned taste aversion paradigm with 15% sucrose as the conditioned stimulus and lithium chloride (LiCl) as the unconditioned stimulus. The results of a dose response experiment yielded evidence that higher doses of LiCl than used in past studies were required to produce profound aversions in these mice, with a 6.0 mEq/kg dose appearing as optimal. When the mice were conditioned with the 6.0 mEq/kg dose in one-bottle tests, there was little evidence of a strain difference in the acquisition of a taste aversion. During extinction trials, however, DBA/2J mice exhibited stronger resistance to extinction than did C57BL/6J mice. Additional analyses of fluid consumption suggested that strain differences in drinking behavior were evident in the paradigm and that these must be considered in interpreting the strain differences observed in conditioned taste aversion. PMID- 6284874 TI - Subclasses of adenosine receptors in brain membranes from adult tissue and from primary cultures of chick embryo. PMID- 6284875 TI - Opiate receptor: multiple effects of metal ions. PMID- 6284876 TI - Characterization of infant rat cerebral cortical membrane proteins phosphorylated in vivo: identification of the ACTH-sensitive phosphoprotein B-50. AB - This study on the phosphorylation in vivo of membrane proteins in cerebral cortices of infant rats reports the identification of the adrenocorticotropin (ACTH)-sensitive phosphoprotein B-50 as one of the substrate proteins that are rapidly phosphorylated in vivo following intracisternal administration of 2 mCi [32P]orthophosphate. Rats were sacrificed 30 min after isotope injection. A fraction enriched in membranes, designated neural membranes (NM), was isolated from the cerebral cortices according to the procedure used for preparation of synaptic plasma membranes (SPM) from adult brain. This NM fraction was characterized by electron microscopy. The proteins of NM were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography. Numerous protein bands of NM in infant rat brain were phosphorylated in vivo. Attention was focussed on the 32P-labeled protein bands in the molecular weight range of 47K-67K. In this region one phosphoprotein band (MW 48K) was more highly labeled than the other bands. The electrophoretic behavior of three of these labeled bands, designated a, c, and e (MW 48K, 55K, and 62K, respectively) was compared with that of protein bands that were phosphorylated in vitro in cerebral membranes isolated from noninjected infant rats. The effects of ACTH1-24 and cyclic AMP in the in vitro system were also studied to probe for the presence of specific membrane proteins known to be sensitive to these modulators. On incubation of NM with [gamma-32P)ATP in the presence and absence of ACTH1-24 in vitro, phosphorylation of a 48K protein band was inhibited in a dose-dependent fashion by the neuropeptide. Two-dimensional electrophoretic separation of NM proteins labeled in vivo indicated that the 48K band had an isoelectric point of 4.5, identical to that of the ACTH-sensitive B-50 protein previously identified. Cyclic AMP stimulated phosphorylation in vitro of two protein bands (MW 55K and 59K) in NM preparations. This result indicates that the in vivo labeled band c may correspond to the cyclic AMP-sensitive 55K protein, whereas phosphoprotein band e, labeled in vivo, appears to be different from the cyclic AMP-sensitive 59K protein band. These observations indicate that neural membranes isolated from infant rat cerebral cortices contain a variety of proteins that can be phosphorylated in vivo. Several of these, for example, the 48K protein band, have the properties of synaptic plasma membrane proteins of adult rat brain that have been characterized by their sensitivity to neuromodulators in endogenous phosphorylating systems in vitro. PMID- 6284877 TI - Adenosine receptor agonists inhibit K+-evoked Ca2+ uptake by rat brain cortical synaptosomes. AB - The uptake of Ca2+ by a K+-depolarized rat brain cerebral cortical crude synaptosomal preparation (P2 fraction) was investigated. The characteristics of the Ca2+ uptake system are similar to those observed by other investigators. The preparation is also a suitable model with which to study the effects of adenosine on Ca2+ uptake and neurotransmitter release, as it is generally accepted that K+ evoked Ca2+ uptake is intimately related to depolarization-induced release of neurotransmitters. We have demonstrated that an extracellular receptor is involved in mediating the adenosine-evoked inhibition of K+-evoked Ca2+ uptake. The pharmacological properties of the receptor suggest that it may be similar in some respects to the A2-receptor associated with adenylate cyclase. The adenosine uptake inhibitor, dipyridamole, potentiated the action of adenosine, suggesting that re-uptake is important in controlling the extracellular adenosine concentration and thus in the regulation of the adenosine receptor. The adenosine receptor antagonist theophylline inhibited the effects of adenosine. Calmodulin inhibited K+-evoked uptake of Ca2+ by the synaptosomal fraction. PMID- 6284879 TI - Demonstration of 2',3'-cyclic nucleotide 3'-phosphohydrolase in cultured human Schwann cells. AB - Schwann cell cultures were established from adult human sural nerve biopsies. 2'3'-Cyclic nucleotide 3'-phosphohydrolase (CNPase) activity was estimated in the homogenates of those cells by a sensitive isotope assay using [3H]2',3'-cyclic AMP as substrate. A high level of CNPase activity was observed in cultured Schwann cells, whereas cultured human muscle and skin fibroblasts contained negligible levels of CNPase activity. CNPase of human Schwann cells followed typical enzyme-substrate kinetics, with an apparent Km of 1.6 mM for 2',3'-cyclic AMP, and the enzyme was stimulated by detergents such as Triton X-100 and deoxycholate. It was inhibited by p-chloromercuricbenzoate and 2'-AMP. These properties are typical of CNPase isolated from adult brain and spinal cord. CNPase can serve as a new biochemical marker of normal cultured human Schwann cells and can be useful in analyzing the properties of cultured Schwann cells from patients with dysschwannian neuropathies. PMID- 6284878 TI - gamma-Aminobutyric acid and benzodiazepine binding sites in audiogenic seizure susceptible mice. AB - The properties of gamma-aminobutyric acid recognition sites, benzodiazepine binding sites and the effect of exogeneous gamma-aminobutyric acid on benzodiazepine binding were determined in crude membrane fractions prepared from the brains of DBA/2 mice at ages before (8-9 and 17-18 days), during (22-23 and 28-29 days) and after (40-43 days) the age of high susceptibility to audiogenic seizures. These have been compared with data from age-matched mice of a strain (TO) with lower audiogenic seizure susceptibility. The number of high-affinity [3H]gamma-aminobutyric acid binding sites was lower at all ages in DBA/2 mice compared with TO mice, but the affinity was higher in DBA/2 mice. The number of low-affinity [3H]gamma-aminobutyric acid binding sites was lower at 8-9 days and 40-43 days in DBA/2 mice, but was not significantly different from TO mice at other ages. For [3H]flunitrazepam binding, the only difference found was a slight reduction in the number of binding sites at 28-29 days of age in DBA/2 mice. gamma-Aminobutyric acid stimulation of [3H]flunitrazepam binding was not significantly different up to 22-23 days of age, but was higher in DBA/2 mice at 28-29 days and lower at 40-43 days. Impairment of gamma-aminobutyric acid function is a possible permissive factor in the age-dependent audiogenic seizure susceptibility in DBA/2 mice. PMID- 6284880 TI - A pharmacological study of two bisbenzylisoquinoline alkaloids, thalistyline and obamegine. AB - Thalistyline, a monoquaternary bisbenzylisoquinoline alkaloid isolated from Thalictrum sp. inhibited respiration in anesthetized dogs. Thalistyline is about one-fourth as potent as d-tubocurarine in blocking neuromuscular transmission in the rat hemidiaphragm preparation. The pharmacological mechanism of action of the alkaloid is similar to that of d-tubocurarine. Obamegine did not exhibit curare like activity. On the isolated rabbit aorta, contractions induced by an alpha adrenoreceptor agonist, phenylephrine, were antagonized by both alkaloids. Increasing concentrations of thalistyline produced parallel shifts to the right in the dose-response curves of phenylephrine. The pA2 value for the competitive pharmacological antagonism was 6.33. Obamegine also antagonized the effects of phenylephrine on the aorta, line and obamegine lowered blood pressure in normotensive dogs. The effect was transient. Repeated injections of the alkaloids resulted in tachyphylaxis to blood pressure lowering effects. Although alkaloids exhibited alpha adrenergic blockade in the vascular preparation, the mechanism for the hypotensive effect remains to be established. PMID- 6284881 TI - Myotonic muscular dystrophy. Calcium-dependent phosphatidate metabolism in the erythrocyte membrane. AB - It has been suggested that the erythrocytes of myotonic dystrophy (MyD) patients have a decreased calcium-stimulated phosphatidic acid (PA) accumulation. This could be the result of a defect in the calcium-stimulated hydrolysis of the polyphosphoinositides (calcium-dependent phosphodiesterase) or in the subsequent formation of PA from its precursors (diacylglycerol kinase). In vitro assays were established for both enzymes in erythrocyte membranes. Calcium-dependent phosphodiesterase activity was assayed with both endogenous 32P-labeled erythrocyte diphosphoinositide and triphosphoinositide and with the same phospholipids isolated from rat brain. No significant differences in activity were found between MyD patients and normal controls with either method of substrate preparation. No difference in diglyceride kinase activity was found between ghosts prepared from MyD patients and normal controls. Thus, there were no differences in either of the membrane-associated enzymes of phosphatidic acid metabolism. PMID- 6284882 TI - Herpes simplex encephalitis. An immunohistological study of the distribution of viral antigen within the brain. AB - An immunoperoxidase technique was used to map the sites of herpes simplex virus antigen (VA) within the brain in 29 autopsied cases of herpes simplex encephalitis. Attention was directed particularly to those parts of the brain that are known from pathological studies to be involved in the disease. Material was studied from cases surviving for varying periods from a few days to a few years after the onset of neurological disease. VA was found within the brain in all cases dying within 3 weeks of onset, but in none dying thereafter. VA was already most abundant in patients dying within the first week and remained plentiful during the first 16 days. Inflammation and necrosis reached a peak when detectable virus was waning. VA was concentrated mainly in the medial and inferior temporal lobes, hippocampus, amygdaloid nuclei, olfactory cortex, insula and cingulate gyrus. It was invariably present on both sides of the brain but was more abundant on one side than the other. Virus was found in glial cells of the olfactory tracts but not in relation to trigeminal pathways. Attention is drawn to focally extensive infection of the granule cell of the dentate fascia, and the possible significance of this finding is discussed. PMID- 6284883 TI - Virus antibodies in Parkinson's disease. Herpes simplex and measles virus antibodies in serum and CSF and their relation to HLA types. AB - Serum and cerebrospinal fluid (CFS) immunoglobulin G (IgG) antibodies to herpes simplex (HSV) and measles viruses were assayed with a radioimmunoassay in 56 patients with idiopathic Parkinson's disease and in a similar number of age- and sex-matched controls with other neurological diseases. As a group, the patients with Parkinson's disease had a significantly increased serum antibody level against HSV, but measles virus antibody levels were similar in both groups. Both in the Parkinson's group and in the control group, the levels of the total IgG in CSF were within normal limits and the CSF antibodies to HSV and measles virus paralleled the serum antibody titers relative to the total IgG serum-to-CSF ratios. This indicates no increased intrathecal antibody production in either group. In 48 patients with Parkinson's disease who were HLA-typed, no association of viral antibody levels with particular HLS antigens were noted. The findings suggest that HSV is not present within the central nervous system of the patients with Parkinson's disease. The increase HSV antibody level seen in Parkinson's disease patients may reflect a more general disturbance of the patients' immune functions. PMID- 6284884 TI - A longitudinal study of antibrain antibodies in multiple sclerosis. AB - The presence of complement-fixing antibrain antibodies is a distinctive feature of multiple sclerosis (MS). In a longitudinal study of 35 MS patients antibrain antibody titres in serum were followed for up to 5 years; in 18 of them also CSF titres were determined. No consistent correlations between antibrain antibody titres and clinical events were found. Thus, MS relapses are not caused by a general increase in antibrain antibody titres, and conversely the relapses did not cause a boosting of antibrain antibodies. Significant variations in the local plaque environment are, however, not ruled out by the present results. PMID- 6284885 TI - Von Recklinghausen's disease with a malignant meningeal, cerebral and optic nerve tumour and bilateral vagal schwannomas. Possible mesenchymal histogenesis on light and electron microscopy. AB - The clinical, histopathologic and fine structural features of multiple unusual tumours detected in a 20-year-old patient with von Recklinghausen's disease, who died within a year of onset of symptoms of a rapidly expanding intracranial tumour, are described. The tumour was found to involve the falx cerebri, the basal leptomeninges and dura mater, both olfactory and optic nerves, both frontal lobes, the right temporal lobe and middle cerebral peduncle, both middle cerebellar peduncles, and with a metastasis in a cervical node. On light and electron microscopy this tumour appeared to be a fibroblastic meningeal sarcoma with giant cells, mitotic figures, a rich reticulin matrix throughout, and tumour cells full of rough ER but without any glial filaments. Also very unusual was the involvement of both vagus nerves in their cervical and intrathoracic portions, by a schwannomatous benign tumour and with a non-chromaffin paraganglioma at its termination in the oesophagus. One of the few cutaneous "neurofibromas" was also schwannian, containing tumour cells with a basement membrane on electron microscopy. Most, if not all, of these tumours appeared mesenchymal in origin, more aggressive in behaviour and carrying a graver prognosis in von Recklinghausen's disease. PMID- 6284886 TI - The effect of postsynaptic block on development of the neuromuscular junction in postnatal rats. AB - The effects of reduced muscle activity on the ultrastructural development of the rat neuromuscular junction (NMJ) have been studied. Soleus muscles of rats were treated with alpha-bungarotoxin (alpha BTx) in order to produce a postsynaptic block of activity between the ages of 10 and 12 days. Muscles of litter mates were treated with saline over the same period. The development of these control and alpha BTx treated muscles was then compared to that of normal muscles from untreated litter mates. During the period between the tenth and twelfth days after birth, normal soleus NMJs show two major ultrastructural changes. 1. The average number of axon terminal profiles present at each endplate decreases. This is thought to reflect the withdrawal of superfluous axons from the endplates. 2. There is an increasing specialization of the postsynaptic structures of the junction: complexity of folding of the muscle junctional membrane increases, as does the accumulation of subjunctional sarcoplasm and muscle nuclei. In soleus muscles treated with alpha BTx, the number of axon terminal profiles observed at the endplates does not decrease, suggesting that elimination of supernumerary axons does not occur. In addition, specialization of the postsynaptic structures of the NMJ is retarded during the period of ACh receptor block. PMID- 6284887 TI - Single channel currents activated by gamma-aminobutyric acid, muscimol, and (-) pentobarbital in cultured mouse spinal neurons. AB - The patch electrode technique was used to record single channel current pulses in tissue-cultured mouse spinal cord neurons. In agreement with earlier noise studies, channels activated by gamma-aminobutyric acid (GABA), muscimol, and (-) pentobarbital were found to have equal unit conductances. The kinetics of channel closing were studied by analyzing the distributions of open state lifetimes. Channels activated by (-)-pentobarbital and muscimol had longer mean open times than channels activated by GABA. As a result, the kinetics of (-)-pentobarbital- and muscimol-activated channels could be studied in greater detail. Most observed open state lifetime distributions were not exponential but contained an excess of short duration events. A sum of two exponential functions gave a much better fit than a single exponential function to most observed open state lifetime distributions. A critical comparison of noise analysis with single channel recording shows that the fast process responsible for the rapid closures would be very difficult to detect in a noise experiment. The channel noise is dominated by the slower process, and as a result, the relaxation time of the slower kinetic component derived from single channel studies is close to the mean open state lifetime derived from noise measurements. The observation of a faster process points toward either an additional population of channels or a scheme for the channel closing transition which is not a simple first order process. PMID- 6284888 TI - Differential regulation of the mu-, delta-, and kappa-opiate receptor subtypes by guanyl nucleotides and metal ions. AB - The effects of Na+, guanyl-5'-yl imidodiphosphate (Gpp(MH)p), and Mn2+ on the binding of dihydromorphine, ethylketocyclazocine, D-Ala2, D-Leu5-enkephalin, and diprenorphine to the opiate receptor were investigated. Three distinct binding sites, mu, delta, and kappa sites, were identified with the use of multiple tracer displacement curves. Moreover, this approach was used to determine the effects of Gpp (NH)p and metal ions on each individual binding site. At the mu and delta sites, Na+ and Gpp(NH)p each decreased and Mn2+ increased agonist binding affinities, with the exception of D-Ala2, D-Leu-enkephalin affinity which was not affected by Gpp(NH)p. None of these conditions markedly altered dihydromorphine and D-Ala2, Leu5-enkephalin binding to kappa sites, whereas the affinity of ethylke-tocyclazocine for kappa sites was decreased by Gpp(NH)p. Sodium ions lowered the capacity of mu sites and Gpp(NH)p reduced that of delta sites, while both agents increased the capacity of apparent kappa sites. These results demonstrate that each of the kinetically distinguishable binding sites is regulated differentially by metal ions and guanyl nucleotides. Simultaneous addition of Na+ and Gpp(NH)p greatly reduced the binding affinity of all three agonists at their respective high affinity sites (dihydromorphine at the mu site, D-Ala2, D-Leu5-enkephalin at the delta site, and ethylketocyclazocine at the kappa and mu sites). This result confirms previous observations that agonist binding is characterized by a large affinity reduction in the presence of both Na+ and guanyl nucleotides, and it extends this concept to each of the opiate receptor subtypes. PMID- 6284889 TI - A comparative study of avermectin B1a and other modulators of the gamma aminobutyric acid receptor . chloride ion channel complex. AB - The interactions of the anthelmintic agent avermectin B1a, the anticonvulsant pentobarbital, and the anxiolytic tracazolate with the gamma-aminobutyric acid (GABA) receptor . chloride ion channel complex in rat brain membrane were studied. The results indicated that they all potentiated ligand binding to the GABA and benzodiazepine receptors. The stimulatory effects of avermectin B1a and pentobarbital, but not tracazolate, on GABA receptor binding were inhibited by picrotoxin. The effect of avermectin B1a was not additive with those of tracazolate and pentobarbital. On the other hand, the stimulatory effect of GABA on benzodiazepine binding was additive with those of avermectin B1a and pentobarbital, but tracazolate and pentobarbital inhibited the effect of avermectin B1a. In the receptor heat inactivation experiments, avermectin B1a and clonazepam protected GABA receptors, whereas avermectin B1a and GABA protected benzodiazepine receptors. Tracazolate, pentobarbital, and picrotoxin did not protect either receptor. These findings suggest that the recognition sites for the benzodiazepines, avermectin B1a, pentobarbital, and picrotoxin are coupled allosterically to the GABA receptor . chloride ion channel complex in different ways. The binding sites for avermectin B1a may be partially shared by picrotoxin, pentobarbital, and tracazolate. PMID- 6284890 TI - Simplified and accurate CT-guided needle biopsy of central nervous system lesions. PMID- 6284891 TI - Effects of dietary cellulose, pectin and oat bran on the small intestine in the rat. AB - The present study was undertaken to determine if three sources of dietary fiber would alter length, weight, DNA, protein or enzyme activity in the small intestine since various fibers are known to decrease intestinal absorption. Rats were fed semipurified diets that contained either 20% cellulose (C), 20% oat bran, 5% pectin (P) or no fiber source (FF). Leucine aminopeptidase activity were significantly greater in the P and C groups when compared with the FF group. There were no significant differences in sucrase activity. Animals in the P group had heavier and longer small intestine and heavier mucosa than the FF group. There were no significant differences in total mucosal DNA or protein. These results indicate that two sources of dietary fiber, cellulose and pectin, can change leucine aminopeptidase activity in the small intestine. PMID- 6284892 TI - Effect of the maternal vitamin D status at parturition on the vitamin D status of the neonatal calf. AB - The plasma concentrations of calcium, phosphorus, magnesium, hydroxyproline, vitamin D, and vitamin D metabolites were determined in cows and their colostrum deprived calves. At birth, calf plasma calcium, phosphorus, and hydroxyproline concentrations were not correlated (P greater than 0.05) with the maternal plasma concentrations of these substances. There was a high degree of correlation between maternal and neonatal calf plasma concentrations of 25 hydroxyergocalciferol (r = 0.733), 25-dydroxycholecalciferol (r = 0.888), 24,25 dihydroxyergocalciferol (r = 0.770), 24,25-dihydroxycholecalciferol (r = 0.629), and 25,26-dihydroxycholecalciferol (r = 0.840). Neonatal calf plasma concentrations of 1,25-dihydroxyvitamin D were low (41.2 +/- 3.4 pg/ml) and had no correlation with maternal concentrations (r = 0.219, P greater than 0.05). Neonatal plasma calcium and inorganic phosphorus concentrations were correlated (P less than 0.05) with maternal plasma concentrations of 1,25-dihydroxyvitamin D (r = 0.559 and 0.525, respectively). Vitamin D status of the dam, therefore, appears to be important in determining neonatal calf plasma concentrations of 25 hydroxyvitamin D, 24,25-dihydroxyvitamin D, and 25,26-dihydroxyvitamin D, and, in addition, the plasma calcium and inorganic phosphorus status of the neonatal calf is apparently dependent on maternal concentrations of 1,25-dihydroxyvitamin D. PMID- 6284893 TI - Effects of dietary fibers on lipid metabolism in liver and adipose tissue in chicks. AB - Two experiments were conducted to investigate the effect of dietary fibers on lipid metabolism with 14-day-old male chicks fed isonitrogenous and isocaloric diets with or without 4% cellulose or 8% alfalfa meal for 3 and 6 weeks. Feeding the cellulose or alfalfa meal resulted in significant reductions in hepatic lipid deposition and plasma lipid content. Hepatic de novo fatty acid synthesis from acetate, glucose and leucine was significantly reduced by feeding the cellulose or alfalfa meal. Triglyceride synthesis from palmitate in the hepatic tissue was significantly reduced by dietary alfalfa meal. Malate dehydrogenase (oxaloacetate decarboxylating) (NADP+) activity in the hepatic tissue was not affected by feeding the cellulose or alfalfa meal for 3 and 6 weeks. Secretion of lipoprotein from the liver appears to be accelerated by the dietary fibers. Lipoprotein lipase activity in the adipose tissue and clearance rate of intravenously injected lipid were enhanced by feeding the alfalfa meal. Feeding the cellulose for 3 weeks reduced hormone-sensitive lipase activity in the adipose tissue. PMID- 6284894 TI - NAD glycohydrolase activity in the liver of rats fed on excess leucine diet and low or high protein diet. AB - In rats fed for 2 weeks on a 10% casein diet supplemented with 5% leucine, the hepatic NAD content was lowered when comparing with the control rats pair-fed on the 10% casein diet. The hepatic NAD glycohydrolase [EC 3.2.2.5] activity increased by 25% in the leucine-supplemented diet-fed rats and by 40% in the nicotinic acid-free diet-fed rats. The hepatic NAD glycohydrolase activity in the rats fed on the nonprotein diet freely for 1 week was elevated by 60% compared to the rats fed on the 18% casein diet. The hepatic NAD content in the former animals was significantly lower than that in the latter. The 70% casein diet caused changes in neither hepatic NAD content nor NAD glycohydrolase activity. These results indicated an inverse relationship between hepatic NAD content and NAD glycohydrolase activity. PMID- 6284895 TI - Crystallographic and structural alterations in the mineral phase of human enamel with carious attacks. AB - The present study aims to obtain further information about the crystallographic and structural alterations in the mineral phase of enamel with the onset of caries. For this purpose, X-ray microbeam diffraction analysis was carried out on ground sections prepared from natural white spot lesions. Electron spin resonance (ESR) analysis was also performed on block samples cut from white spot lesions and from undamaged enamel of the same teeth. The results of crystallinity measurements showed that enamel apatite in demineralized lesions was lower in crystallinity than the apatite in the surface layer and surrounding sound enamel. This X-ray diffraction study also revealed the presence of two non-apatitic minerals in the demineralized lesions. One type of the mineral is whitlockite, giving spotty rings. The nature of the other mineral, giving a single ring-like reflection, remains to be elucidated. A comparison of ESR spectra taken from the caries-attacked and undamaged enamel provided evidence that the former samples have a lower degree of alignment of apatite microcrystals. The occurrence of non apatitic mineral phases and the observed difference in microcrystalline alignment may be the results of a remineralization process. PMID- 6284896 TI - Carbonate-derivative centers in x-ray irradiated carbonate-containing apatites. PMID- 6284897 TI - Plasma exchange in two "receptor negative" homozygotes for autosomal, dominant hypercholesterolemia. PMID- 6284898 TI - [Experimental inner ear pathology with herpes simplex virus infection. I. Intracerebral inoculation of herpes simplex virus in mice]. PMID- 6284899 TI - Natural history of parainfluenza virus infection in childhood. AB - In order to determine the natural history of parainfluenza virus infection in early life, we followed prospectively 130 infants and children from birth or a few months of age for evidence of infection with PV. Using rapid diagnostic techniques, standard tissue culture infectivity, and serologic techniques we were able to document primary PV infection in 92% of these infants, and repeated infection with heterotypic or homotypic PV strains in 49% by 30 months of age. Increasing patient age had no significant effect in reducing the incidence of lower respiratory tract illness as a result of PV infection. Infection with one PV serotype provided no protection against LRTI at the time of subsequent infection with a heterotypic PV strain. In contrast, primary PV infection provided a brief period of immunity to LRTI upon homotypic reinfection. Secretory IgA responses to PV were determined by immunofluorescent techniques. Antibody response to PV strains causing primary infection and heterotypic repeated infection were transient and of low magnitude. Homotypic reinfection resulted in significantly enhanced production of secretory antibody to PV. At least in early life, repeated exposures to PV appear to be essential for maintaining immunity to severe forms of illness caused by PV infection. PMID- 6284900 TI - Usefulness of bone marrow imaging in childhood malignancies. AB - Two hundred six technetium-99m sulfur colloid bone marrow scans in 110 pediatrics patients were reviewed. The normal distribution of sulfur colloid in the lower extremities in various age groups was established. There was progressive loss of uptake with increasing age from less than two years to greater than ten years. Tumor replacement was seen as regions of decreased radioactivity, and the extent of the scan defect paralleled the response of the disease to therapy. Both chemotherapy and irradiation resulted in an extension of the Tc-99m SC to peripheral marrow sites. In irradiated areas, marrow scan defects were demonstrated and generally recovered normal activity by six months after the completion of therapy. Marrow scan abnormalities caused by tumor replacement were present in four patients despite normal bone scans and radiographs. Ultimate confirmation of tumor involvement was by needle aspiration or biopsy. Persistent marrow defects were seen in two patients with neuroblastoma who had remission of their disease: biopsy revealed myelofibrosis. Technetium-99m sulfur colloid bone marrow scanning is a sensitive monitor of altered marrow activity associated with pediatric hematologic or oncologic diseases. PMID- 6284901 TI - Gianotti disease or Gianotti-Crosti syndrome? PMID- 6284903 TI - Rotavirus: a cause of nosocomial infection in the nursery. PMID- 6284902 TI - A possible association between hepatitis-B antigen-negative infantile papular acrodermatitis and Epstein-Barr virus infection. PMID- 6284904 TI - The efficacy of corticotropin in primary infantile spasms. PMID- 6284905 TI - Purification and properties of NADP-linked, alcohol dehydrogenase from Entamoeba histolytica. AB - An NADP-linked, alcohol dehydrogenase from Entamoeba histolytica was purified to apparent homogeneity by Blue Sepharose affinity chromatography. Molecular weights of 130,000 and 30,000 were estimated by gel filtration and by sodium dodecyl sulfate polyacrylamide gel electrophoresis, respectively, suggesting that the enzyme is a tetramer. The enzyme exhibited more than 20-fold selectivity for NADP(H) over NAD(H). Although the purified enzyme acts on both primary and secondary alcohols, higher activity was found with secondary alcohols. PMID- 6284906 TI - Mouse IgE response to an allergen from Entamoeba histolytica. AB - Attempts were made to demonstrate the presence of allergenic components in an extract of axenically cultured Entamoeba histolytica (HK 9). Various strains of mice were immunized i.p. with a low dose of the extract mixed with Al(OH)3 and boosted with the same mixture 4 wk later. A high titer of IgE antibody response was demonstrated in BALB/c, DBA/2 (H-2d), CBA (H-2k), and SWM strains but not in C3H/He (H-2k) strain. The extract was fractionated by gel filtration with Sephadex G-200 and then by DEAE-cellulose chromatography with a gradient buffer with progressive increase in molarity. Fractions having the strongest activity to stimulate the IgE antibody response in the mouse were subjected to SDS polyacrylamide gel electrophoresis. From the results of gel filtration and polyacrylamide gel electrophoresis, the molecular weight on the partially purified allergen was estimated to be 25,000 to 27,000 daltons. PMID- 6284907 TI - Human gingival tissues in culture synthesize three metalloproteinases and a metalloproteinase inhibitor. PMID- 6284908 TI - Analysis of conjugated estrogens in a vaginal cream formulation by capillary gas chromatography. AB - A capillary gas-chromatographic method is described for the quantitative analysis of nine equine estrogens in a vaginal cream formulation. The sodium sulfate ethers of the estrogens were selectively extracted from the formulation, subjected to enzyme hydrolysis, and derivatized to their oxime-trimethylsilyl esters. Resolution of the resulting derivatives was achieved on short (15 m) capillary column, wall-coated with cyanopropylmethyl silicon stationary phase. PMID- 6284910 TI - Studies on the mechanism of nitrofurantoin-mediated red cell toxicity. AB - The mechanism of nitrofurantoin-mediated depletion of red cell reduced glutathione (GSH) was investigated. Nitrofurantoin caused cellular depletion of GSH in vitro under aerobic and oxygen-depleted conditions, an effect which could be partially inhibited by coincubation with the hemeprotein ligand ethyl isocyanide, or completely prevented by coincubation with 2'-AMP, an inhibitor of NADPH-dependent reductase enzymes. Covalent binding of nitrofurantoin to red cell macromolecules appeared to be a minor process and was not substantially inhibited by either ethyl isocyanide or 2'-AMP. Covalent binding was only slightly greater under oxygen-depleted conditions. Nitrofurantoin increased the rate of superoxide formation in red cell lysate, an effect inhibited by ethyl isocyanide but not by 2'-AMP. These data suggest different mechanisms for nitrofurantoin-mediated depletion of GSH under aerobic and oxygen-depleted conditions. In the presence of oxygen, nitrofurantoin causes the release of superoxide from oxyhemoglobin. The superoxide thus formed may deplete GSH via several mechanisms. In the absence of oxygen, nitrofurantoin is reduced to reactive metabolites via reactions which appear to require the participation of both an NADPH-dependent flavoprotein and hemoglobin. PMID- 6284909 TI - Effects of indomethacin on beta adrenoreceptor-stimulated renin release in the dog. AB - The hypothesis that prostaglandins participate in beta adrenoreceptor-stimulated renin release was tested by examining the effects of prostaglandin synthesis inhibition on renal nerve stimulation (0.5-5.0 Hz)-induced renin release. In phentolamine (5 micrograms/kg/min, intrarenal artery)-treated dogs (n = 6), renal nerve stimulation produced a frequency-related increase in renal renin secretion rate, without altering renal blood flow. In phentolamine plus propranolol (0.3 mg/kg i.v. bolus + 5 micrograms/kg/min infusion)-treated dogs (n = 6), renal nerve stimulation-induced renin release was inhibited by 86% (P less than .03), 75% (P less than .03) and 73% (P less than .02) at 1, 2 and 5 Hz, respectively. In contrast to propranolol, blockade of prostaglandin synthesis with indomethacin (8 mg/kg i.v. bolus) failed to alter the renin secretory response to renal nerve stimulation in phentolamine-treated animals. These results indicate that, in a situation of intrarenal alpha adrenoreceptor blockade, renal nerve stimulation induces renin secretion by a beta adrenoreceptor mechanism that is independent of prostaglandin biosynthesis. PMID- 6284911 TI - Modulation of dopamine release in rat striatal slices by delta opiate agonists. AB - The effects of various opiates on the spontaneous release of [3H]dopamine ([3H]DA) continuously formed from [3H]tyrosine has been studied in rat striatal slices. Morphine (5 x 10(-6) M), fentanyl (5 x 10(-8) M) and the tripeptide Tyr-D Ala-Gly-NH-CH(CH3)CH2-CH(CH3)2 (10(-6) M) were without effect, whereas D-Ala2-Met enkephalinamide (3 x 10(-7) to 10(-5) M), D-Ala2-D-Leu2-enkephalin (5 x 10(-9) M) and the hexapeptide Tyr-D-Ser-Gly-Phe-Leu-Thr (10(-7) M) enhanced [3H]DA release in vitro. The D-Ala2-Met-enkephalinamide stimulation of [3H]DA release was not accompanied by any increase in [3H]DA synthesis and persisted in the presence of tetrodotoxin (5 x 10(-7) M). Naloxone (5 x 10(-7) M) completely blocked the effect of Tyr-D-Ser-Gly-Phe-Leu-Thre (10(-7) M) on [3H]DA release. However, the opiate antagonist did not affect the action of the hexapeptide or of D-Ala2-Met enkephalinamide when used at a concentration equal to or lower than the agonist. This suggests that both peptides act on opiate receptors having a low sensitivity to naloxone. According to these various results and to the pharmacological characteristics of the opiates tested as described in peripheral organs or in the brain, it is concluded that opiates acting on delta opiate receptors may presynaptically regulate the release of DA in the striatum. PMID- 6284912 TI - Effect of sodium orthovanadate on renal renin secretion in vivo. AB - The effect of vanadate (0.5 mumol/min) on renin secretory rate (RSR) of the kidney has been studied in nembutal-anesthetized, volume-expanded dogs. Intrarenal vanadate infusion caused a 69.3 +/- 8.8% decrease in RSR. This was accompanied by marked decreases in renal blood flow (RBF), glomerular filtration rate (GFR) and fractional excretion of sodium (FENa). Renal vascular resistance rose from 1.3 +/- 0.09 to 6.1 +/- 2.3 mm Hg/ml/min (P less than .0005). Papaverine infusion partially blunted the effect of vanadate on RSR (RSR only fell to 42. +/- 10% of basal values). The decreases in RBF and GFR were also less and FENa slightly higher than normal. Acetylcholine prevented the effects of vanadate more fully. There was no fall in RBF, GFR or FENa and it basically abolished the fall in RSR which fell only 19.4 +/- 25.3 of control (P = N.S.). Nifedipine (a slow Ca++ channels blocker) also prevented the fall in RBF, GFR and FENa induced by vanadate. RSR did not change significantly (7.8 +/- 10.9%). These results clearly demonstrate that vanadate is a potent inhibitor of renin secretion and suggest that inhibition of smooth muscle Na+, K+, adenasine triphosphatase and changes in the cystosolic concentration of Na and Ca are involved in its mechanism. Changes in perfusion pressure and sodium delivery to the macula densa appear to have little if any role in the inhibition. PMID- 6284913 TI - Inhibition of acetylcholine responses by intracellular calcium in Lymnaea stagnalis neurones. AB - 1. Acetylcholine (ACh)-induced currents were studied in completely isolated Lymnaea stagnalis neurones using the voltage-clamp technique. 2. The ACh activated pathways were shown to be selective for Cl- ions. 3. It was shown that membrane depolarization inhibits ACh-induced conductance. This phenomenon was called 'ACh response inactivation'. 4. Inactivation decreases after lowering the extracellular Ca2+ concentration or after blockade by Mn2+ of the electrically excitable Ca2+ channels. 5. In dialysed neurones an increase of the intracellular Ca2+ concentration inhibits the ACh-induced conductance. 6. The conclusion is made that the inactivation of ACh response by depolarization is initiated by Ca2+ entering the neurone through the electrically excitable Ca channels. 7. The onset and the decay of the ACh response inactivation were studied by analysing the relaxations of the ACh-induced current during and after the application of depolarizing pulses. The most conspicuous relaxation is a slow relaxation observed at the end of a long depolarizing pulse, which appears to reflect the return of the system from the inactivated state to the non-inactivated one. 8. The slow relaxations observed during and after a depolarizing pulse appear correlated with variations of the intracellular Ca2+ concentration, and are distinct from faster relaxations observed in the hyperpolarizing range and attributed to the voltage dependence of the channel open-time. PMID- 6284914 TI - Gallamine triethiodide-induced modifications of sodium conductance in Myxicola giant axons. AB - 1. Internal gallamine triethiodide (Flaxedil) modifies Na+ channel kinetics in Myxicola axons but does not alter K+ conductance. The drug has no effect externally. 2. Gallamine initially increases the leakage-conductance, but this effect completely reverses within 30 min despite the maintained presence of drug. 3. During step depolarizations to membrane potentials less than -10 mV, gallamine slows the rate of Na+ inactivation, but all channels which have opened can still inactivate. During depolarizations to more positive potentials, gallamine modified Na+ currents show a biphasic decline, and at VM greater than -10 mV, Na+ inactivation is incomplete as evidenced by the large Na+ tail currents which follow pulses sufficiently long to have allowed complete inactivation of normal Na+ channels. The tail currents are slower than normal Na+ tails, and exhibit a pronounced hook. With gallamine, the fraction of Na+ channels which do not inactivate increases sigmoidally over the range 0 mV to +80 mV. 4. For VM greater than ENa, gallamine almost completely blocks outward Na+ currents. The block is determined by the direction of Na+ current, rather than the absolute membrane potential. 5. Gallamine has no effect upon the rate of Na+ channel activation, the maximum Na+ conductance, the steady-state Na+ inactivation curve, or the rate of development or removal of inactivation by prepulses. 6. Gallamine eliminated physiological immobilization of intramembrane charge movements (QOFF and QON) and does not itself induce immobilization. Thus, in the presence of gallamine, QOFF following long pulses is the same as QOFF following short pulses. PMID- 6284917 TI - The role of the pituitary-adrenocortical axis in reflex responses of the adrenal medulla of the dog. AB - 1. The release of catecholamines from the adrenal medulla, in response to carotid body hypoxia, may outlast the stimulus by more than 30 min. 2. After denervation of the adrenal gland the immediate release of catecholamines in response to carotid hypoxia is abolished, but the prolonged release remains. 3. The prolonged release of catecholamines is abolished by cycloheximide. 4. Both corticotrophin in vivo and hydrocortisone in the isolated perfused adrenal gland release adrenomedullary catecholamines. 5. It is concluded that a component of the response of the adrenal medulla to carotid body hypoxia is mediated by corticotrophin and corticosteroid release. PMID- 6284915 TI - Role of presynaptic calcium ions and channels in synaptic facilitation and depression at the squid giant synapse. AB - 1. The roles of presynaptic calcium influx and calcium accumulation in synaptic facilitation and depression were explored at the giant synapse in the stellate ganglion of the squid. 2. Calcium currents were recorded in the presynaptic terminal, using a three-electrode voltage clamp and blocking sodium and potassium currents pharmacologically. The calcium influx was constant during pairs or trains of brief depolarizing pulses that elicited facilitating or depressing excitatory post-synaptic potentials (e.p.s.p.s). 3. The relationship between calcium influx and transmitter release during brief depolarizing pulses of varying amplitude resembled a power function with exponent of about 2. 4. Presynaptic calcium concentration transients were measured by injecting the dye arsenazo III and detecting absorbance changes microspectrophotometrically. Increments in intracellular free calcium accompanying single action potentials appeared constant for repeated action potentials that elicited facilitating e.p.s.p.s. 5. The presynaptic calcium concentration remains elevated for several seconds following action potentials. 6. Presynaptic injection of calcium ions by interbarrel ionophoresis evokes a postsynaptic depolarization, apparently reflecting a large increase in miniature e.p.s.p. frequency. Presynaptic action potentials remain unaffected by this treatment, but e.p.s.p.s triggered by them are facilitated for several seconds, and then depressed. 7. The results are consistent with the hypothesis that synaptic facilitation is due to the action of residual calcium or a calcium complex remaining in the presynaptic terminal after electrical activity. The late depression of release during calcium injection may be a result of the continual release of transmitter and consequent depletion of a presynaptic store. PMID- 6284916 TI - Changes in holding and ion-channel currents during activation of an ascidian egg under voltage clamp. AB - 1. The unfertilized egg of an ascidian, Halocynthia roretzi, was activated by the divalent ionophore A23187 in natural or artificial sea water (nSW or ASW) or by an external solution containing a high concentration of Ca ions (high-Ca ASW) under voltage-clamp condition.2. Activation current began with an abrupt increase in the holding current and decayed relatively slowly with a common time course in various ASWs. Activation current was both Na- and Ca-dependent. The peak time, T(p), and the peak amplitude, D, of the activation current in nSW at 15 degrees C and -90 mV were 27 +/- 4 sec and -1.50 +/- 0.47 nA, respectively.3. The currents through Na, Ca and anomalous K channels were evoked by test pulses with constant intervals in nSW and high-Sr, high-Ca and high-K ASWs. Na-channel current was enhanced during activation. In contrast, Ca-channel current decreased. In high Ca, Na-free ASW the Ca current through Na channels increased while the Ca current through Ca channels decreased. The time for the maximum of Na current, T(max), was (7.2 +/- 1.5) x 10 sec at 15 degrees C and index R, the ratio of the maximum amplitude to the amplitude before activation, was 2.31 +/- 0.18 in nSW. The time for the minimum of Ca-channel current, T(min), was about 70 sec, being almost the same as T(max) of Na current.4. The current through anomalous K channels increased initially and decreased later with a time lag behind the decrease in Ca channel current.5. Both T(p) of activation current and T(max) of Na current were reduced by raising the temperature. Q(10) for T(p) and T(max) was 2.2 and 2.3, respectively.6. When the egg was activated in ASW containing scorpion toxin, the Na current through normal channels increased strongly while the Na current through toxinmodified channels increased less markedly.7. There was no significant change in the total membrane capacity before and during activation. PMID- 6284918 TI - Active groups of saxitoxin and tetrodotoxin as deduced from actions of saxitoxin analogues on frog muscle and squid axon. AB - 1. The actions of three saxitoxin (STX) analogues have been studied on the frog sartorius muscle fibre and the squid giant axon. One--neosaxitoxin--is a natural analogue, and two--decarbamylsaxitoxin and reduced saxitoxin--are synthetic. 2. The maximum dV/dt of the action potential in paired-muscle protocol is reduced by the analogues with relative potencies: STX (1), tetrodotoxin (1), neo-STX (1), decarbamyl-STX (0.2) and reduced-STX (0.01). 3. In constant-current studies on frog muscle fibres and in voltage-clamp studies on squid axons, all three analogues block only the sodium channel without affecting the potassium channel. 4. All three analogues bind to the same site as does STX in a competitive manner. 5. The experimental results suggest that the active groups in STX are the 7,8,9 guanidinium and the C-12 hydroxy groups. The carbamyl group contributes to, but is not essential for activity. 6. Stereospecific groups in the tetrodotoxin (TTX) molecule are the 1,2,3 guanidinium and the C-9, C-10 hydroxy groups. C-4 and C-8 groups are also important. 7. As new view is proposed in which STX and TTX can bind to a receptor located in the outside surface of the membrane very close to the orifice of the sodium channel. PMID- 6284919 TI - Calcium and potassium currents in muscle fibres of an insect (Carausius morosus). AB - 1. A three electrode voltage-clamp was used to investigate membrane currents in the skeletal muscle fibres of the stick insect, Carausius morosus. Contraction was blocked by hypertonic solutions. 2. Membrane currents elicited by step depolarizations consisted of an inward current, an early outward current and a delayed outward current. 3. The reversal potential of the delayed outward current did not change when SO4(2-) was substituted for Cl-, but shifted by 14.1 mV when [K]0 was increased from 20 mM to 40 mM in SO4(2-) solution, suggesting that the delayed current is carried by K+. Both early and delayed outward currents were substantially reduced by 120 mM-tetraethylammonium (TEA) ions. 4. The small size of the shift in the reversal potential of the delayed outward current with increased pulse duration suggests that the delayed current measured flows mainly across the surface membrane. 5. Increasing [Ca]o made the apparent reversal potential for the inward current (120 mM-TEA Ringer) more positive and increased the size of the maximum inward current. However, Ca-currents showed saturation with increasing [Ca]o, indicating that there is a site to which Ca ions bind during their passage through the membrane. The dissociation constant of this site was 7.3 mM at 0 mV and was voltage-dependent. 6. Inward currents were blocked by 1 mM-La3+ or Cd2+, or by substitution of Co2+ or Ni2+ for Mg2+. Strontium and barium were able to permeate the channel but Na+ and Mg2+ appear impermeant. 7. As expected from the low intracellular Ca concentration, the instantaneous current-voltage relation of the Ca current rectified strongly in the inward direction. 8. Both constant field theory and the simplest, single site, Eyring rate theory model predict the rectification of the instantaneous current-voltage relation. The rate theory model also predicts saturation of the Ca current with [Ca]o. PMID- 6284920 TI - The control of liver regeneration after partial hepatectomy in the rat. AB - 1. Normal rat liver slices were incubated for 24 h in culture medium to which had been added freeze-dried plasma, prepared from the blood of rats which had undergone partial hepatectomy (60-70% of liver removed) 10 min, 4 or 24 h previously. No changes in the dry weight of the slices were observed. 2. Similarly, there were no changes in the dry weight or protein content of incubated liver slices when freeze-dried plasma prepared from the blood of rats unilaterally nephrectomized 10 min, 4 or 24 h previously was added to the culture medium. 3. Slices from the remaining liver stump, taken 4 h after partial hepatectomy (60-70% of liver removed) and incubated for 4 h in culture medium, did not release a factor capable of increasing the dry weight of normal liver slices incubated in the same medium. 4. After partial hepatectomy (60-70% of liver removed), there was no increase of guanosine 3':5'-cyclic monophosphate in the remaining part of the liver. 5. No measurable amount of esterase activity was found in normal liver or in the remaining liver stump after partial hepatectomy (60-70% of liver removed). 6. It is suggested that the stimulatory mechanism for liver regeneration is of a different nature from that involved in compensatory renal hypertrophy. PMID- 6284921 TI - Electrophysiological studies of new-born rat nodose neurones in cell culture. AB - 1. Neurones of the nodose ganglion of the vagus nerve were dissociated from new born rats and grown in the virtual absence of non-neuronal cells and in the presence of nerve growth factor. 2. The resting potentials of the neurones ranged from -40 to -80 mV. Action potentials were of short duration, with no inflexion on the falling phase; others were of longer duration with a hump on the falling phase. 3. The inward current of the action potential was carried either predominantly by Na+ or by Na+ and Ca2+. 4. Tetrodotoxin (1 microM) blocked the Na+ channels of some neurones but in other neurones the Na+ channels were partially or completely resistant to tetrodotoxin (1-10 microM). 5. Many neurones formed excitatory synapses on neighbouring neurones which were blocked or greatly reduced by conventional ganglionic nicotinic antagonists. This indicates that these neurones secreted ACh and expressed ACh receptors at these synapses. 6. The accompanying paper (Baccaglini & Cooper, 1981) reports the effect of co-culturing nodose neurones with non-neuronal cells on the expression of functional nicotinic receptors. PMID- 6284922 TI - Nucleoside translocation in sheep reticulocytes and fetal erythrocytes: a proposed model for the nucleoside transporter. AB - 1. Nucleoside transport by fetal erythrocytes from nucleoside-permeable and nucleoside-impermeable type new-born lambs and by reticulocytes from adult sheep was compared with that of mature erythrocytes from adult sheep of the two phenotypes.2. Fetal cells and reticulocytes transported [U-(14)C]uridine rapidly, with little difference between cells from the two types of sheep. Transport occurred by a saturable uptake mechanism with similar properties to that present in mature cells from adult nucleoside-permeable type animals, except for an approximately 100-fold higher V(max).3. This increased translocation capacity was associated with increased numbers of high-affinity [(3)H]nitrobenzylthioinosine binding sites ( approximately 2000-3000 sites/cell compared with approximately 20 sites/cell for mature nucleoside-permeable sheep erythrocytes).4. The calculated transport capacity for each nucleoside translocation site is therefore similar in all cell types (140-180 molecules/site. s at 25 degrees C, assuming that each transport site binds a single molecule of inhibitor). These values compare favourably with turnover estimates for the nucleoside transporter from human and pig erythrocytes.5. Loss of nucleoside transport activity after birth closely paralleled loss of [(3)H]nitrobenzylthioinosine binding sites and the progressive loss of fetal cells from the circulation. Similarly, reticulocyte maturation in vitro was also associated with rapid loss of both nucleoside transport capacity and inhibitor binding activity.6. p-Chloromercuriphenylsulphonate and trypsin had no effect on [(3)H]nitrobenzylthioinosine binding to intact fetal cells. In contrast, both agents markedly inhibited binding to isolated ;ghosts' where both sides of the cell membrane were accessible to reagent. p Chloromercuriphenylsulphonate inhibition was markedly reduced in the presence of uridine, and reversed by addition of dithiothreitol.7. We conclude that nucleoside transport changes during ontogeny and reticulocyte maturation in the sheep as well as species differences in nucleoside transport capacity are regulated by variations in the numbers of functional transport sites per cell rather than by changes in the activity of a constant number of sites. It is also likely that the nucleoside carrier exhibits chemical asymmetry.8. A simple molecular model of the erythrocyte nucleoside transporter consistent with these and other known properties of the carrier is proposed. PMID- 6284924 TI - [Electrophysiology of neurosecretory cell system (author's transl)]. PMID- 6284923 TI - Interactions between three slow potassium responses controlled by three distinct receptors in Aplysia neurones. AB - 1. A voltage clamp study was made of the K(+) permeability increases produced in certain identifiable neurones of the cerebral ganglion of Aplysia by any one of three distinct agonists (carbachol, histamine and dopamine).2. The three K(+) responses involve three distinct receptors, as shown by the selective effects of reversible antagonists (Gruol & Weinreich, 1979a) as well as by the differential, irreversible effects of trimethyloxonium (TMO) ions.3. Prolonged exposure of the neurones to one of the agonists reduces the response to the same agonist (desensitization) and, over the low concentration range, doubling the concentration of an agonist leads to supra-linear summation (potentiation).4. Prolonged exposure of the neurones to one of the agonists also reduces the response to the other agonists (cross-desensitization) and combined application of two agonists reveals cross-potentiation.5. The time course of desensitization (onset and decay) was the same for the histamine and carbachol responses and, except at very high concentration, was indistinguishable from that of cross desensitization. Likewise, potentiation was similar in the two agonist systems and did not differ significantly from cross-potentiation.6. The results can be interpreted by assuming that the responses to the three agonists involve specific steps followed by common reaction steps, and that some of the common reaction steps control both potentiation and desensitization.7. The responses to carbachol and histamine differ in their voltage sensitivity. This suggests that one or more of the specific steps are voltage-sensitive.8. Although an increase of the intracellular Ca(2+) concentration can itself open K(+) channels, and also inhibit the responses to the three agonists, an increase of internal Ca(2+) does not appear to play an important role either in the development of the response or in the desensitization process. PMID- 6284925 TI - Electrophoretic analysis of endonuclease-generated fragments of k-DNA, of esterase isoenzymes, and of surface proteins as aids for species identification of insect trypanosomatids. AB - In order to verify the applicability of biochemical methods for species identification of Trypanosomatidae, 13 species of monoxenic trypanosomatids plus the heteroxenous Trypanosoma cruzi were comparatively analyzed by three different biochemical methods. Insect trypanosomatids examined were: Crithidia acanthocephali, C. fasciculata (three varieties), C. luciliae luciliae, C. luciliae thermophila, C. deanei, C. oncopelti, Herpetomonas muscarum muscarum, H. megaseliae, H. samuelpessoai, H. mariadeanei, Leptomonas seymouri, L. collosoma, L. samueli, and Blastocrithidia culicis. Also included in the survey were aposymbiotic strains of C. deanei and C. oncopelti. Methods used were: electrophoretic profiling of endonuclease-generated fragments of k-DNA, esterase isoenzymes profiling, and polyacrylamide-gel electrophoresis (SDS-PAGE) of radioiodinated cell surface proteins. Interspecific but not intraspecific differences were detected by all three methods among the 13 monoxenic species examined. Thus, it is concluded that these methods can be successfully used, in addition to classical criteria, for species identification of insect trypanosomatids. PMID- 6284926 TI - Identification and characterization of growth hormone receptors on isolated rat adipocytes. AB - It is now recognized that growth hormones (GH) can exert direct metabolic effects on adipose tissue from hypophysectomized or normal rats in vitro. This implies the presence of receptors for GH in this tissue. The interaction of 125I-labelled human and bovine GH has been studied therefore in isolated adipocytes from male and female rats. Binding of 125I-hGH was dependent on time (with equilibrium being reached by 2 hrs at 23 degrees C) and number of fat cells (binding being linear up to 1.5 x 10(6) cells/tube). Binding was partially reversible (half-time for dissociation being 4 hrs at 23 degrees C) and specific for GH being displaced by various growth hormones but not by lactogens or other hormones. Scatchard analysis revealed linear plots with an affinity (KA) of 0.8 x 10(9) M-1 and 24,000 sites per cell for adipocytes from 36-day old male rats. 125I-labeled bovine GH binding was somewhat lower, being reflected in a slightly reduced binding capacity. No obvious sex difference in hGH binding was evident but there was a quite striking age-dependence of binding, being low prior to weaning, reaching a peak between 24 - 36 days and then declining to low levels again in the adult. The demonstration of specific GH receptors in adipose tissue confirms the notion of direct actions of GH in this tissue and should now permit further studies of the relationship between GH binding and GH action. PMID- 6284927 TI - Factors affecting the distribution and stability of unoccupied 1,25 dihydroxyvitamin D3 receptors. AB - Recently our laboratory reported that unoccupied 1,25(OH)2D3 receptors are found in the nuclei/chromatin fraction under low salt conditions in vitro (J. Biol. Chem. 255, 6799-6805, 1980). Additionally, various conditions exert differential effects on receptor solubilization and stability in vitro, potentially leading to confusion when these effects are not fully appreciated. In order to facilitate future studies, we have complied a review of these factors and their effects. The conditions discussed include dilution, ionic strength (KCl and molybdate), protease inhibitors (Trasylol, PMSF and TPCK), temperature, ligand, glycerol, dithiothreitol, and EDTA. PMID- 6284928 TI - Lymphocyte membrane adenosine receptors coupled to adenylate cyclase: properties and occurrence in various lymphocyte subclasses. AB - Lymphocytes possess membrane adenosine receptors through which this nucleoside or analogs activate adenylate cyclase. Several adenosine analogs - 5'-N ethylcarboxamide adenosine (NECA), N6-(L-2-phenyl-isopropyl) adenosine (PIA), 2 chloro adenosine (Cl-Ado) - were tested on the adenylate cyclase of mouse thymocyte homogenates; the order of potencies: NECA greater than Cl-Ado greater than PIA is consistent with the order which has been assumed to be characteristic of Ra type adenosine receptors. The stimulation of thymocyte adenylate cyclase by NECA was biphasic while the responses to Cl-Ado and PIA were not, which suggests the existence of Ra sub-types of adenosine receptors. Moreover, these adenosine receptors were found to be present in several lymphocyte subclasses: mature and immature mouse thymocytes, B and T mouse splenocytes, human peripheral blood lymphocytes, and also in lymphoblastoid cell lines. No crucial differences were observed between the adenylate cyclase stimulations of the various population by NECA. PMID- 6284929 TI - Interaction between iodothyronines and thyrotropin receptor in human cultured thyroid cells. AB - In order to verify the existence of a "short-loop" negative feedback between iodothyronines and adenylate cyclase system of human thyroid, we have studied the effect of preincubation with iodothyronines, iodotyrosines, iodothyronine analogues and iodide on TSH-induced cAMP cellular accumulation in normal human thyroid cells in primary culture. Iodide did not produce an inhibitory effect on TSH-dependent adenylate cyclase system both in normal human thyroid plasma membranes and cultured cells. Iodothyronines at a 30-40 microM concentration did not inhibit the TSH-dependent adenylate cyclase activity of human thyroid plasma membranes; however at a 1 microM concentration they were able to inhibit the TSH dependent cAMP accumulation by cultured cells. Preincubation with iodotyrosines and iodothyronine analogues failed to inhibit the TSH-responsive cAMP accumulation in human thyroid cultured cells. PMID- 6284930 TI - Kinetic characterization of [125I] iodo-prolactin in binding to primary monolayer cultures of rabbit mammary epithelium. AB - Monocellular suspensions of epithelial cells from mammary glands of rabbits at 20 22 days of pregnancy were prepared by sequential dissociation with collagenase hyaluronidase followed by Pronase. Maintenance in D-valine-substituted minimum essential medium (D-valine-MEM) supplemented with 10% dialyzed calf serum yielded monolayers enriched for rabbit mammary epithelial cells (RMEC). RMEC specifically and reversibly bound bovine PRL with Ka = 1.41-1.85 x 10(9)M-1. Association of lactogen with RMEC receptor followed bimolecular reaction kinetics with rate of 5.17 (+/- 0.75) x 10(5)M-1 sec-1 at 24 C, and 1.03 (+/- 0.11) x 10(6)M-1 sec-1 at 37 C. Dissociation was first order (K-1 = 5.97 (+/- 0.70) x 10(-5) sec-1) and was unaffected by the presence of lactogen. Specific binding determined with an excess of unlabelled bPRL was 66-77% of the total binding, and was optimal at pH 7.4. The binding reaction reached equilibrium in 2 h at 37 C, in 3 h at 24 C, and after 24 h at 4 C. Studies of binding capacity revealed the presence of 4.6-6.3 x 10(3) sites per cell, competition for which was limited to hormones demonstrating lactogenic activity. Recovered lactogen was not degraded by incubation with or dissociation from RMEC. Approximately 25% of the radioactivity remained associated with the cells even upon prolonged incubation. These studies demonstrated several advantages of RMEC for the investigation of hormone-receptor interaction and receptor regulation. PMID- 6284931 TI - Endogenous prolactin maintains its own binding sites in the pigeon crop sac mucosa. AB - The specific binding of lactoperoxidase-labelled 125I-labelled ovine prolactin was determined in a membrane particulate of the pigeon crop-sac mucosal epithelium. Binding was found to be dependent upon the particulate preparation used, its protein concentration and the length of the incubation at 5 degrees C. Scatchard analysis of the binding to crop-sacs from saline or prolactin-injected (1.9 microgram per pigeon) revealed that prolactin stimulated 7-fold its own receptors by increasing the number of binding sites per mg protein: saline - 392 +/- 75 fmol/mg protein and prolactin 2736 +/- 602 fmol/mg protein (p less than 0.01). This increase did not affect the affinity constant (Ka): saline - 5.28 +/- 0.75 x 10(8) l/mol and prolactin 3.28 +/- 0.40 x 10(8) l/mol (N.S.), in keeping with the stimulatory effect of prolactin in the rat liver and mammary glands. This study further demonstrates the physiological role of endogenous prolactin in maintaining its own binding-sites in the pigeon crop-sac, since the administration of 0.8 ml anti-serum to prolactin resulted in a 63% reduction in the specific binding of labelled hormone in vitro. These results confirm the prolactin binding to the pigeon crop-sac mucosa, quantify the stimulation of this binding by prolactin itself, and demonstrate the role of the endogenous hormone in the maintenance of these receptors. PMID- 6284932 TI - Rat sperm enzymes during epididymal transit. AB - The activities of cAMP and cGMP phosphodiesterases (EC 3.1.4.1), adenylate cyclase (EC 4.6.1.1) and protein carboxyl-methylase (EC 2.1.1.24) were measured in the particulate and soluble (105 000 g supernatant) fractions of washed spermatozoa isolated from five segments of the adult rat epididymis. The activities of both phosphodiesterases decreased during epididymal transit, whereas adenylate cyclase and protein carboxyl-methylase underwent a progressive increase, the latter showing the most marked alteration. Both cAMP and cGMP phosphodiesterases as well as the adenylate cyclase were all associated primarily with the particulate fraction, and the extent to which these enzymes were associated with the membranes increased as the spermatozoa passed through the epididymis. Sperm protein carboxyl-methylase activity was, on the other hand, predominantly soluble in all segments of the epididymis. Adenylate cyclase, cAMP phosphodiesterase and protein carboxyl-methylase activities were found predominantly in the sperm tails, whereas cGMP phosphodiesterase was equally distributed between heads and tails. These observations imply that the acknowledged increase in intracellular cAMP levels which occurs in spermatozoa during epididymal transit may be a consequence of both increased synthesis (adenylate cyclase) and reduced hydrolysis (phosphodiesterase). PMID- 6284933 TI - Variation in the steroidogenic responsiveness of isolated rat Leydig cells. AB - Isolated Leydig cells were prepared from adult rat testes by (1) mechanical dissection, (2) collagenase dispersion or (3) mechanical dissection followed by collagenase dispersion, and their functional characteristics were assessed. Compared with Methods 2 and 3, mechanical isolation alone resulted in the purest preparation of Leydig cells but the lowest yield. Leydig cells isolated by any of the 3 methods had similar numbers of LH- and LH-RH receptors, but cells isolated by Methods 1 and 3 showed poor testosterone responsiveness compared to cells isolated by Method 2. This reduced response was evident following stimulation with hCG, dibutyryl cyclic AMP or an LH-RH agonist, and could not be accounted for simply on the grounds of diminished cell viability. It is concluded that Leydig cells in the rat testis are particularly sensitive to mechanical intrusion, and this is an important factor to bear in mind when preparing Leydig cells or when comparing results between laboratories. PMID- 6284934 TI - Neuroendocrine control of milk ejection. PMID- 6284935 TI - Extragenital homologous mixed mullerian tumor. PMID- 6284936 TI - The efficacy and safety of radiosynovectomy. PMID- 6284937 TI - Species- or isozyme-specific enzyme inhibitors. 4. Design of a two-site inhibitor of adenylate kinase with isozyme selectivity. PMID- 6284938 TI - Synthesis, X-ray crystallographic determination, and opioid activity of erythro-5 methylmethadone enantiomers. Evidence which suggests that mu and delta opioid receptors possess different stereochemical requirements. AB - Enantiomers of erythro-5-methylmethadone (3) were synthesized from optical antipodes of erythro-3-(dimethyl-amino)-2-butanol. X-ray crystallographic analysis of (-)-3 perchlorate revealed that it possesses the 5S,6S absolute configuration. It was found that (-)-3 is substantially more potent than its enantiomer (+)-3 as an opioid agonist in vivo and in vitro. In vitro tests (guinea pig ileal longitudinal muscle and mouse vas deferens preparations) suggest that (-)-3 mediates its effect chiefly through mu opioid receptors. On the other hand, (+)-3 and the more potent enantiomers of methadone, (-)-1, and isomethadone, (-)-2, appear to have less mu-receptor selectivity and interact with a greater fraction of delta receptors than does (-)-3. The fact that the solid-state conformation of (-)-3 differs from that of (-)-1 and (-)-2, which show great similarity in conformational features, suggests that mu and delta receptors have different conformational requirements. The possibility of different modes of interaction with a single opioid receptor population also is discussed. PMID- 6284939 TI - Sodium currents in the giant axon of the crab Carcinus maenas. AB - Measurements were made of the kinetics and steady-state properties of the sodium conductance changes in the giant axon of the crab Carcinus maenas. The conductance measurements were made in the presence of small concentrations of tetrodotoxin and as much electrical compensation as possible in order to minimize errors caused by the series resistance. After an initial delay of 10-150 microsec, the conductance increase during depolarizing voltage clamp pulses followed the Hodgkin-Huxley kinetics. Values of the time constant for the activation of the sodium conductance lay on a bell-shaped curve with a maximum under 180 microsec at -40 mV (at 18 degrees C). Values of the time constant for the inactivation of the sodium conductance were also fitted using a bell-shaped curve with a maximum under 7 msec at -70 mV. The effects of membrane potential on the fraction of Na channels available for activation studied using double pulse protocols suggest that hyperpolarizing potentials more negative than -100 mV lock a fraction of the Na channels in a closed conformation. PMID- 6284940 TI - Potassium currents in the giant axon of the crab Carcinus maenas. AB - Measurements were made of the kinetic and steady-state characteristics of the potassium conductance in the giant axon of the crab Carcinus maenas. These measurements were made in the presence of tetrodotoxin, using the feedback amplifier concept introduced by Dodge and Frankenhaeuser (J. Physiol, (London) 143:76-90). The conductance increase during depolarizing voltage-clamp pulses was analyzed assuming that two separate potassium channels exist in these axons. The first potassium channel exhibited activation and fast inactivation gating which could be fitted using the m3h, Hodgkin-Huxley formalism. The second potassium channel exhibited the standard n4 Hodgkin-Huxley kinetics. These two postulated channels are blocked by internal application of caesium, tetraethylammonium and sodium ions. External application of 4 amino-pyridine also blocks these channels. PMID- 6284941 TI - Mechanism of calcium release from skeletal sarcoplasmic reticulum. AB - Ca2+ -induced Ca2+ release at the terminal cisternae of skeletal sarcoplasmic reticulum was demonstrated using heavy sarcoplasmic reticulum vesicles. Ca2+ release was observed at 10 mum Ca2+ in the presence of 1.25 mm free Mg2+ and was sensitive to low concentrations of ruthenium red and was partially inhibited by valinomycin. These results suggest that the Ca2+ -induced Ca2+ release is electrogenic and that an inside negative membrane potential created by the Ca2+ flux opens a second channel that releases Ca2+. Results in support of this formulation were obtained by applying a Cl- gradient or K+ gradient to sarcoplasmic reticulum vesicles to initiate Ca2+ release. Based on experiments the following hypothesis for the excitation-contraction coupling of skeletal muscle was formulated. On excitation, small amounts of Ca2+ enter from the transverse tubule and interact with a Ca2+ enter from the transverse tubule and interact with a Ca2+ receptor at the terminal cisternae and cause Ca2+ release (Ca2+ -induced Ca2+ release). This Ca2+ flux generates an inside negative membrane potential which opens voltage-gated Ca2+ channels (membrane potential dependent Ca2+ release) in amounts sufficient for contraction. PMID- 6284943 TI - Ion selectivity of the apical membrane Na channel in the toad urinary bladder. AB - The ion selectivity of the apical membrane Na channel in the toad urinary bladder was investigated. The electrical potential difference and resistance across the basal-lateral membrane were reduced using high concentrations of KCl in the serosal bathing medium, and gradients for various ions were imposed across the apical membrane by altering the composition of the mucosal bathing medium. Ion fluxes through the channel were measured as the transepithelial current inhibited by amiloride, a specific blocker of the channel's Na conductance. The selectivity sequence for alkali metal cations was H greater than Li greater than Na much greater than K. K permeability was barely detectable; the selectivity for Na over K was about 1000:1. Ammonium, hydroxyl ammonium and hydrazinium ions were, like K, virtually impermeant. The results suggest that the size of the unhydrated ion is an important factor in determining permeability in this channel. PMID- 6284942 TI - Interaction of spin-labeled local anesthetics with the sodium channel of squid axon membranes. AB - The effects of spin-labeled local anesthetics on sodium currents of internally perfused squid axons were studied using the voltage-clamp technique. Internal application (10 mum) of the most potent spin-labeled local anesthetic used in this study produced a small initial block of sodium currents. However, after sixty repetitive pulses (to + 80 mV) given at 1 Hz, the sodium currents were drastically reduced. In addition to this frequency-dependent phenomenon, the anesthetic effect on the sodium currents was also sensitive to the voltage of the pulses. Both the frequency- and voltage-dependent properties remained intact after removal of sodium inactivation with pronase. The recovery of sodium currents from this frequency-dependent anesthetic effect followed a single exponential curve with a surprisingly long time constant of about 10 min. Such a long recovery time, which is longer than any known sodium inactivation process, led us to suggest that the recovery process represents the dissociation of drug molecules from their binding sites. We have also found that increasing hydrophobic character of the homologues series of spin-labeled local anesthetics enhances the frequency- and voltage-dependent block of sodium currents. This effect strongly suggests that hydrophobic interaction is an integral component of the binding site. These probes with their selective effects on the sodium currents, are expected to be highly useful in studying the molecular structure of the sodium channels. PMID- 6284944 TI - Effect of monensin on osmotic water flow across the toad bladder and its stimulation by vasopressin and cyclic AMP. AB - The effects of the sodium ionophore monensin on osmotic water flow across the urinary bladder of the toad Bufo marinus were studied. Monensin alone did not alter osmotic water flow; however, the ionophore inhibited the hydrosmotic response to vasopressin and cyclic AMP in a dose-dependent manner. The inhibitory effects of monensin were apparent when the ionophore was added to th serosal bathing solution but not when it was added to the mucosal bathing solution. The inhibitory effect of serosal monensin required the presence of sodium in the serosal bathing solution but not the presence of calcium in the bathing solutions. Thus, it appears that intracellular sodium concentration is a regulator of the magnitude of the hydrosmotic response to vasopressin and cyclic AMP. PMID- 6284945 TI - Cerebellar afferents to paramedian lobule from the trigeminal complex in Tupaia glis: a horseradish peroxidase (HRP) study. AB - Projections from the trigeminal complex to paramedian lobule (PML) were studied in the tree shrew (Tupaia glis) by means of retrograde transport of horseradish peroxidase (HRP). Neurons which project to both dorsal and ventral folia of PML are located primarily in those areas of the trigeminal nuclear complex interpreted as nucleus interpolaris (Vi) and caudal areas of the nucleus oralis (Vo). The majority of HRP-labeled neurons lie in ventral and ventrolateral regions of Vi/Vo. No HRP-reactive cells are present in the principal (Vp), mesencephalic, or motor nuclei nor in nucleus caudalis or rostral portions of oralis. The majority of trigeminocerebellar (TC) cells are found in ipsilateral Vi; however, sparse numbers of labeled somata are present in this subnucleus on the contralateral side. Within Vi/Vo, small fusiform and medium-and large-sized multipolar neurons contain HRP-reaction product. Large multipolar cells are found primarily in ventrolateral portions of Vi/Vo, while medium and small neurons are scattered throughout the ventral half of the nucleus. Small-sized neurons are also present dorsally within Vi/Vo. Axons of labeled TC cells course laterally through the spinal trigeminal tract, enter medial aspects of the restiform body, and arch dorsally into the cerebellum. PMID- 6284946 TI - Biases of the estimates of DNA divergence obtained by the restriction enzyme technique. AB - A mathematical formula for the relationship between the average number of nucleotide substitutions per site and the proportion of shared restriction sites between two homologous nucleons is developed by taking into account the unequal rates of substitution among different pairs of nucleotides. Using this formula, the possible amount of bias of the estimate of the number of nucleotide substitutions obtained by the Upholt-Nei-Li formula for restriction site data is investigated. The results obtained indicate that the bias depends upon the nucleotides in the recognition sequence of the restriction enzyme used, the unequal rates of substitution among different nucleotides, and the unequal nucleotide frequencies, but the primary factor is the unequal rates of nucleotide substitution. The amount of bias is generally larger for four-base enzymes than for six-base enzymes. However, when many restriction enzymes are used for the study of DNA divergence, the bias is unlikely to be very large unless the rate of substitution greatly varies from nucleotide to nucleotide. PMID- 6284947 TI - Binding of nickel (II) to 5'-nucleoside monophosphates and related compounds. AB - The interactions of Ni(II) cation with a representative suite of purine bases and the respective nucleosides and nucleotides have been studied by ultraviolet difference spectroscopy. Apparent association constants, Kapp, were determined for each system at pH 7.0, using computer linear regression coupled with an iteration technique. The specificity of binding of Ni2+ for the purine nucleotides studied at pH 7.0 was 5'-GMP greater than 5'-IMP greater than 5'-AMP; a similar ordering was also found for the respective nucleosides and bases. In this study binding was not observed for the suite of pyramidines used, although a Ni2+ - cytidine complex has been observed (Fiskin and Beer, 1965). It was also found that Ni2+ bound more strongly to the purine 5'-nucleotides than to the respective nucleosides and bases. These trends are explained in terms of metal ligand bonds and available bonding positions on the ligands. A role for metal-ion nucleotide types of complexes is suggested in the processes that might have given rise to the origin of life. PMID- 6284949 TI - Purification of polybrominated biphenyl congeners. AB - A commercial preparation of polybrominated biphenyls (Firemaster BP-6) was fractionated by preferential acetone solubilization and a number of PBB congeners were purified from the resulting fractions by repeated recrystallization, alumina adsorption column chromatography, and reversed-phase Lipidex-500 column chromatography. The applicability of reversed-phase Lipidex-5000 chromatography for the separation of highly structurally similar brominated biphenyl congeners that otherwise could not be separated by alumina chromatography or recrystallization was demonstrated. In total, seven congeners were purified, including one whose structure was assigned as 2,3,4,5,3',4'-hexabromobiphenyl. PMID- 6284948 TI - Mitochondrial DNA sequences of primates: tempo and mode of evolution. AB - We cloned and sequenced a segment of mitochondrial DNA from human, chimpanzee, gorilla, orangutan, and gibbon. This segment is 896 bp in length, contains the genes for three transfer RNAs and parts of two proteins, and is homologous in all 5 primates. The 5 sequences differ from one another by base substitutions at 283 positions and by a deletion of one base pair. The sequence differences range from 9 to 19% among species, in agreement with estimates from cleavage map comparisons, thus confirming that the rate of mtDNA evolution in primates is 5 to 10 times higher than in nuclear DNA. The most striking new finding to emerge from these comparisons is that transitions greatly outnumber transversions. Ninety-two percent of the differences among the most closely related species (human, chimpanzee, and gorilla) are transitions. For pairs of species with longer divergence times, the observed percentage of transitions falls until, in the case of comparisons between primates and non-primates, it reaches a value of 45. The time dependence is probably due to obliteration of the record of transitions by multiple substitutions at the same nucleotide site. This finding illustrates the importance of choosing closely related species for analysis of evolutionary process. The remarkable bias toward transitions in mtDNA evolution necessitates the revision of equations that correct for multiple substitutions at the same site. With revised equations, we calculated the incidence of silent and replacement substitutions in the two protein-coding genes. The silent substitution rate is 4 to 6 times higher than the replacement rate, indicating strong functional constraints at replacement sites. Moreover, the silent rate for these two genes is about 10% per million years, a value 10 times higher than the silent rate for the nuclear genes studied so far. In addition, the mean substitution rate in the three mitochondrial tRNA genes is at least 100 times higher than in nuclear tRNA genes. Finally, genealogical analysis of the sequence differences supports the view that the human lineage branched off only slightly before the gorilla and chimpanzee lineages diverged and strengthens the hypothesis that humans are more related to gorillas and chimpanzees than is the orangutan. PMID- 6284950 TI - Effects of phospholipase A2 inhibitors on diethyl maleate-induced lipid peroxidation and cellular injury in isolated rat hepatocytes. AB - Isolated hepatocytes provide a suitable system for investigation of various aspects of the mechanism of a toxic response. The mechanism by which most chemicals induce hepatotoxicity is still not known. Evidence that phospholipases may play a role in cellular injury has been reported. In the present study the effects of reported inhibitors of phospholipase A2 (quinacrine, chlorpromazine, dexamethasone, and dibutyryl cyclic AMP) on diethyl maleate (DEM)-induced lipid peroxidation, reduced glutathione (GSH) depletion, and cellular injury were examined in isolated hepatocyte suspensions. Hepatocytes were incubated for 7 h under control conditions or with (1) DEM (4 mM), (2) one of the inhibitors (quinacrine, 10, 50, or 150 microM; chlorpromazine, 50 microM; dexamethasone, 0.1, 0.5, 1, or 2.5 mM; dibutyryl cyclic AMP, 0.1, 0.5, 1, or 2.5 mM) or aspirin (500 microM), or (3) a combination of DEM and one of the inhibitors or aspirin to determine their effect on DEM toxicity. Samples were withdrawn at hourly intervals for estimation of cellular injury (loss of intracellular K+ and lactate dehydrogenase and trypan blue exclusion index), lipid peroxidation (thiobarbituric acid reactants assay), and GSH concentration. Quinacrine and chlorpromazine inhibited DEM-induced lipid peroxidation but not cellular injury or GSH loss. This suggests that phospholipase A2 may be involved in DEM-induced lipid peroxidation but not cell damage. However, dexamethasone and dibutyryl cyclic AMP enhanced both lipid peroxidation and loss of cell viability due to DEM, suggesting novel aspects of the biochemical mechanisms of chemically induced cytotoxicity. PMID- 6284951 TI - Decrease in the number of lower affinity (type II) nerve growth factor receptors on embryonic sensory neurons does not affect fiber outgrowth. AB - Nerve growth factor binds to two different specific receptors on responsive cells. The relationship of these two receptors is not fully understood at this time. We have studied the binding of labeled NGF to a different strain of white leghorn chicken embryo dorsal root ganglionic cells. The equilibrium dissociation constants for the two sites (KdI = 4.1 +/- 1.8 x 10(-11) M, KdII = 1.0 +/- 0.8 x 10(-9) M) are identical to those obtained previously. Also, the number of type I sites per cell (3.8 +/- 1.3 x 10(3)) is the same as that previously determined. However, the number of type II sites per cell (1.9 +/- 1.3 x 10(4)) is significantly different than that previously determined. This 2.5-fold decrease in the number of type II sites does not affect the concentration of NGF needed to obtain maximal fiber outgrowth from explanted sensory ganglia. The rate of association (1.2 +/- 0.2 x 10(7) M-1 sec-1 at 22 degrees C) of labeled NGF with receptors on sensory neurons from this different strain of chickens is identical to that previously obtained. The rate of association of NGF with its receptors on sensory neurons was also determined at 4 degrees C. This rate constant (2.1 +/- 1.1 x 10(6) M-1 sec-1) along with the rate constants obtained at 22 degrees and 37 degrees C were used to determine an activation energy for the binding of NGF to its receptors. The activation energy obtained (16.2 kcal/mole) suggests that binding is not a diffusion-controlled process. PMID- 6284952 TI - Effects of methadone on ornithine decarboxylase and cyclic nucleotide phosphohydrolase in neuronal and glial cell cultures. AB - Mixed neuronal and nonneuronal cell cultures were obtained from 8-day-old chick embryos cerebral hemispheres and glial-enriched cultures were obtained from fifteen-day-old chick embryo cerebral hemispheres. Cultures were exposed to methadone, a narcotic drug, from days four to six. The activity of ornithine decarboxylase (ODC) was determined at day eight and the activity of cyclic nucleotide phosphohydrolase (CNP) was determined at day fifteen. Both ODC and CNP activity were higher in mixed neuronal-nonneuronal cell cultures treated with methadone as compared to control. No effect was observed in the neuronal-enriched or glial-enriched cultures. These findings are interpreted to reflect that neuronal-glial interaction is important in the response of primary neural cells to methadone. PMID- 6284953 TI - Changes in enzyme activities of glycerolipid metabolism of guinea-pig cerebral hemispheres during experimental hypoxia. AB - Hypoxic treatment causes changes of some enzymatic activities involved in the glycerolipid metabolism in subcellular fractions of guinea pig cerebral hemispheres. The activity of lysophosphatidylcholine acyltransferase, choline phosphotransferase, glycerol-3-phosphate acyltransferase(s), as well as the activity of triacylglycerol lipase significantly decreased in the microsomes of cerebral hemispheres of animals intermittently exposed to hypoxic treatment for eighty hours. At the same time, a marked activation of microsomal and mitochondrial phospholipase A2 occurred. The changes of the above-mentioned enzymatic activities after the hypoxic treatment correlated well with the increase in the level of brain and blood free fatty acids. The changes also correlated with the decrease of labeled lipid precursors incorporated into lipids of the cerebral hemispheres, observed during oxygen insufficiency. PMID- 6284954 TI - Neisseria gonorrhoeae: a modified medium for cultivation and primary isolation in the absence of gaseous carbon dioxide. PMID- 6284955 TI - The pharmacokinetics and tissue levels of polymyxin B, colistin and gentamicin in calves. AB - Following a single intravenous injection of polymyxin B, colistin (5 mg/kg, each) and gentamicin (3 mg/kg) to calves, the decline in serum antibiotic concentration generally suggested a three-compartment (open system) pharmacokinetic model. Tissue binding is a dominant factor in the distribution and elimination kinetics of the drugs. Less than 65% of the dose of polymyxin B and colistin was recovered in the urine during 48 h after treatment. Concentrations of nonbound polymyxin B and colistin in the kidney, liver, lung, heart, and skeletal muscles were similar to total (free and bound) serum drug levels, but considerably higher concentrations were found, in bound form, in chloroform-ethanol extracts of these organs. At 24 h after treatment, more than 50% of the doses of polymyxin B and colistin were present bound to the tissues; the largest amount was in the skeletal muscles. Gentamicin was concentrated in the kidney, predominantly in the free form. At 48 h after treatment the amount of gentamicin in the kidney was 6.3% of the administered dose, being more than five times greater than the corresponding amounts of polymyxin B and colistin. The extent of tissue uptake of polymyxin B and colistin limits the usefulness of kinetic values, which are derived from the analysis of serum drug levels, for the purpose of designing dosage schedules. The strong affinity of the polymyxins to the muscle tissue, and gentamicin to the kidney, can result in drug residues persisting in the body for several weeks. PMID- 6284956 TI - Stability of polyoma DNA sequences and virus-coded proteins during tumor formation. AB - To determine the stability of polyoma viral DNA in transformed rat cells during their growth in vivo, we compared the state and arrangement of polyoma virus DNA sequences in virus-transformed rat cell lines before and after their passage in vivo. In cell lines from 12 independent tumors induced by the inoculation of animals with three different transformed cell lines, we could detect no significant changes in the arrangement of viral DNA sequences associated with the in vivo passage of these cell lines. In 13 of 14 tumor cell lines examined, the pattern of polyoma virus tumor antigens, characterized by the presence of the polyoma virus large, middle, and small tumor antigens, was unchanged. PMID- 6284957 TI - Methylation and amplification of mouse mammary tumor virus DNA in normal, premalignant, and malignant cells of GR/A mice. AB - The methylation and amplification of mouse mammary tumor virus (MuMTV) proviral DNA was investigated in normal, premalignant, and malignant tissues of GR/A mice. The proviral methylation pattern was examined with the restriction enzyme HhaI, which fails to cleave methylated DNA. MuMTV proviral DNA from liver, kidney, and heart was highly methylated. Proviral DNA was somewhat undermethylated in mammary gland cells from virgin and lactating mice and extensively undermethylated in cells from premalignant outgrowths, pregnancy-dependent tumors, and pregnancy independent tumors. The restriction enzyme SacI was used to detect additional proviruses in the same cells. No additional proviral copies of MuMTV were detected in liver, kidney, or heart cells or in mammary gland cells from virgin mice. Some mammary gland cells from lactating mice appeared to contain additional copies of the endogenous, highly oncogenic GT-MTV-2 provirus. Premalignant outgrowth, pregnancy-dependent tumor, and pregnancy-independent tumor cells contained an average of two to three additional copies per cell of the GT-MTV-2 provirus. Thus, neoplasia in GR/A mice was directly associated with quantized increases in MuMTV proviral DNA undermethylation and GR-MTV-2 proviral DNA amplification. Restriction enzyme analysis suggested that premalignant outgrowths and pregnancy-dependent tumors both consisted largely of heterogenous cell populations, although some evidence of clonal dominance was detected. PMID- 6284958 TI - Mutation causing premature termination of the polyoma virus medium T antigen blocks cell transformation. AB - We used site-specific mutagenesis to introduce a termination codon, TGA, into the reading frame for the polyoma virus medium T antigen. We induced this mutation in a region of the polyoma genome in which the overlapping coding regions for the large and medium TE antigens are translated in different reading frames. Therefore, the mutation terminated translation of the medium T antigen, but it caused only a single amino acid substitution in the large T antigen and did not affect the small T antigen. Cells infected by the mutant virus produced normal size small and large T antigens. The infected cells produced a 28,000-dalton fragment of the 48,000-dalton medium T antigen, whose size and tryptic peptide map were consistent with its being a truncated N-terminal fragment terminating at the new termination codon of the mutant. Immunoprecipitates of mutant-infected cell extracts did not show medium-T-antigen-associated protein kinase activity. The mutant virus replicated normally in mouse 3T6 cells and induced cellular DNA synthesis in resting mouse 3T3 cells, but it failed to transform rat or hamster cells, as judged by focus formation and growth in agar. The mutant complemented a tsA mutant which affects the large T antigen for transformation, implying that the mutant defect for transformation was in the medium T antigen. These results imply that the small T antigen and the large T antigen together are insufficient to cause transformation and support the conclusion that the medium T antigen is essential for cell transformation by polyoma virus. PMID- 6284959 TI - Simian virus 40 mutant with transposed T-antigen and VP1 genes. AB - A simian virus 40 mutant with the T-antigen gene transposed to the late region of the viral genome has been constructed. This transposed molecule directed the synthesis of a full-sized, 92,000-dalton large T antigen in both permissive and nonpermissive cells. This large T antigen functioned in the initiation of viral DNA replication and in the transformation of nonpermissive cells. T-antigen synthesis by this transposed genome had the characteristics of late transcription, thus indicating that functional large T antigen of simian virus 40 is not required for the initiation of late transcription. PMID- 6284960 TI - MB78, a virulent bacteriophage of Salmonella typhimurium. AB - The isolation and some properties of a virulent bacteriophage of Salmonella typhimurium, MB78, which is morphologically, serologically, and physiologically unrelated to P22, are reported. The phage has a noncontractile long tail with partite ends. It cannot multiply in minimal medium in the presence of citrate. MB78-infected cells are, however, killed in such medium. This phage cannot grow in rifampin-resistant mutants of the host. The latent period of growth of this phage is much shorter than that of P22. Both sieA and sieB genes of the resident P22 prophage are required to exclude the superinfecting MB78 phage, whereas all temperate phages related to P22 are excluded by either one or both of the genes individually. Restriction endonuclease cleavage patterns of P22 and MB78 are distinctly different. The absence of homology between the two phages P22 and MB78 suggests that MB78 is not related to phage P22. PMID- 6284961 TI - In vitro reassembly of vesicular stomatitis virus skeletons. AB - Vesicular stomatitis virus (VSV) has been disrupted with nonionic detergent plus 0.5 M NaCl under conditions which result in solubilization of the viral glycoprotein (G), matrix protein (M), and lipids, leaving the nucleocapsid in a highly extended state. Dialysis of these suspensions to remove NaCl was found to result in reassociation of nucleocapsids with M protein. Reassociated structures were highly condensed and similar in appearance to "native" VSV skeletons produced by extraction of virions with detergent at low ionic strength. For instance, electron microscopic analysis revealed that, like "native" skeletons, "reassembled" skeletons were cylindrical in shape, with diameters in the range of 51.0 to 55.0 nm and cross-striations spaced approximately 6.0 nm apart along the length of the structure. Like native skeletons, reassembled skeletons were found by sodium dodecyl sulfate-polyacrylamide gel electrophoresis to contain the viral N and M proteins, but they lacked the glycoprotein entirely. Both native and reassembled skeletons were found to be capable of in vitro RNA-dependent RNA synthesis (transcription). In vivo skeleton assembly required the presence of M protein and nucleocapsids. No skeleton-like structures were formed by dialysis of nucleocapsids in the absence of M protein or of M protein in the absence of nucleocapsids. These results provide strong support for the view that the VSV M protein plays a functional role in condensing the viral nucleocapsid in vitro and raise the possibility that it may play a similar role in vivo. PMID- 6284962 TI - Small t protein of simian virus 40 is required for dense focus formation in a rat cell line. AB - Incomplete foci were formed in a Fischer rat cell line infected with simian virus 40 detection mutants encoding defective small t antigen. The foci were small and rarely piled up. The fraction of DNA-synthesizing cells in the foci was about half of that induced by wild-type virus. PMID- 6284963 TI - Isolation and preliminary characterization of temperature-sensitive mutants of poliovirus type 1. AB - We isolated six temperature-sensitive mutants of poliovirus type 1 (Mahoney) by hydroxylamine mutagenesis and replica plating at 31, 33 (permissive), and 39 degrees C (restrictive). One of these mutants, designated tsB9, was chosen for more detailed examination. tsB9 accumulated 25% of the wild-type amount of virus specific RNA at the restrictive temperature. We found that tsB9 was not able to synthesize mature, 35S single-stranded RNA at the restrictive temperature. In spite of the absence of significant RNA synthesis, tsB9 retained the ability to inhibit host protein synthesis during infection at 39 degrees C at about the same rate as wild-type virus. PMID- 6284964 TI - Characterization of a polypeptide present in herpes simplex virus type 2 transformed and -infected hamster embryo cells. AB - Transformation of hamster embryo cells by herpes simplex virus stimulated the production of a 35-kilodalton (35K) protein that was specifically immunoprecipitated, along with other polypeptides, by rabbit hyperimmune serum. This 35K polypeptide was further analyzed by partial digestion with Staphylococcus aureus V8 protease in parallel with a 35K polypeptide from herpes simplex virus type 2-infected cells. These polypeptides had almost identical partial-proteolytic cleavage maps, indicating that they are probably the same or that they are very similar polypeptides. PMID- 6284965 TI - Purification of glycoprotein gD of herpes simplex virus types 1 and 2 by use of monoclonal antibody. AB - Glycoproteins gD-1 and gD-2 of herpes simplex virus types 1 and 2, respectively, were purified on an immunoadsorbent consisting of the type-common monoclonal antibody HD-1 linked to Sepharose. Each glycoprotein was of sufficient purity, quantity, and biological activity to be used for immunological and biochemical studies. Each glycoprotein induced high titers of type-common monospecific neutralizing antibody in mice. Amino aicd analysis indicated that gD-1 and gD-2 had similar though not identical amino acid compositions. PMID- 6284966 TI - Effect of the arginine analog canavanine on the synthesis of Epstein-Barr virus induced proteins in superinfected Raji cells. AB - The addition of canavanine to cultures of superinfected Raji cells in the absence of arginine prevented the appearance of early antigens as defined by immunofluorescent staining. Addition of various amounts of arginine permitted the identification of at least three groups of proteins, each responding differently to the various concentrations of arginine-canavanine. PMID- 6284967 TI - Isolation and biochemical characterization of partially transformation-defective mutants of avian myelocytomatosis virus strain MC29: localization of the mutation to the myc domain of the 110,000-dalton gag-myc polyprotein. AB - Recently, we isolated three mutants of MC29 virus which, although able to transform fibroblasts with the same efficiency as wild-type MC29, were 100-fold less efficient at transforming macrophages. In this study we found that MC29 transformed quail producer cell line Q10 was able to generate these partially transformation defective mutants at a high frequency. Using tryptic peptide mapping, we determined that the smaller gag-myc polyproteins encoded by the transformation-defective viruses had lost myc-specific tryptic peptides. This suggested that the mutations which resulted in the transformation-defective viruses being inefficient at transforming macrophages were located in the v-myc sequence and thus directly implicated v-myc and the gag-myc polyprotein in transformation by MC29. PMID- 6284968 TI - Deletions within the transformation-specific RNA sequences of acute leukemia virus MC29 give rise to partially transformation-defective mutants. AB - The viral RNAs of three nonconditional mutants of avian myelocytomatosis virus MC29 were analyzed. These mutants, which were originally isolated from the quail producer line Q10 and were designated 10A, 10C, and 10H, have lost most of the ability to transform hematopoietic cells in vitro and to induce tumors in vivo, but they still transform cultured fibroblasts with the same efficiency as wild type (wt) MC29. Electrophoretic analyses showed that the mutant genomic RNAs were smaller than the 5.7-kilobase genome of wt MC29; the genomes of mutants 10A, 10C, and 10H were about 5.5, 5.3, and 5.1 kilobases long, respectively. Analyses of the transformation-specific sequences of these mutant RNAs by a combination of T(1) oligonucleotide fingerprinting and hybridization with cDNA from the transformation-specific sequences myc of wt MC29 or competition hybridization including wt MC29 RNA revealed that deletions of myc-specific sequences had occurred. The deletions in all three mutants overlapped, since they all had lost one particular myc-specific oligonucleotide. In agreement with the size of the genomic RNAs, mutants 10C and 10H had lost two additional myc oligonucleotides, and mutant 10A contained a modified myc oligonucleotide. The locations of the deletions were deduced from comparisons with previously established oligonucleotide maps of several members of the MC29 subgroup of acute leukemia viruses and by hybridization of wt and mutant RNAs to molecularly cloned subgenomic fragments of wt MC29 proviral DNA, representing the 5' and 3' domains of the myc sequence. We found that the deleted sequences represented overlapping internal segments of the myc sequence and that the borders of myc with the partial complements of the virion genes gag and env appeared to be conserved in mutant and wt MC29 RNAs. The correlation between the altered transforming potential for hematopoietic cells and the partial deletion of myc in the mutant RNAs provided direct genetic evidence for the involvement of myc in oncogenesis. However, the unaffected efficiency of these mutants in fibroblast transformation suggested that the deleted sequences are not essential for the fibroblast transforming potential of the onc gene of MC29. PMID- 6284969 TI - Expression of the PRC II avian sarcoma virus genome. AB - We found that the genomic RNA of the replication-defective avian sarcoma virus PRC II was 4.0 kilobases long. A Northern blot analysis of the viral RNAs present in PRC II-transformed cells showed that the PRC II genome was expressed as a single 4.0 kilobase mRNA species. In vitro translation of polyadenylic acid containing 70S virion RNA yielded two highly related proteins of 110,000 and 105,000 daltons (P110 and P105), which were synthesized from messenger activity that sedimented as expected for the 4.0 kilobase PRC II genome (at 25 to 27S). P110 and P105 were identified as in vitro translation products of the PRC II genome by immunoprecipitation and tryptic peptide mapping and were the only PRC II-specific polypeptides detected by in vitro synthesis. In addition, we found that immune complexes prepared from PRC II 70S virion RNA in vitro translation products contained a tyrosine-specific protein kinase activity. A comparison of the in vitro- and in vivo-synthesized proteins revealed that PRC II-transformed cells also contained 110,000- and 105,000-dalton proteins, which were indistinguishable from in vitro-synthesized P110 and P105 by electrophoretic mobility and tryptic peptide analysis. Both P110 and P105 were present in producer cells and in seven individual nonproducer clones. A pulse-chase analysis showed that P105 was the primary translation product of the PRC II genome and that P110 was derived from P105 by post-translational modification. Under conditions of long-term labeling with [35S]methionine, P110 and P105 were present in a molar ratio of approximately 1:1. These results indicated that the transformation-specific product of the PRC II genome, previously referred to as a single component (P105), actually consists of two polypeptides related by post translational modification. PMID- 6284971 TI - Poliovirus empty capsid morphogenesis: evidence for conformational differences between self- and extract-assembled empty capsids. AB - In this paper we describe the use of specific proteinases, surface-specific radioiodination, and antigenic reactivity in conjunction with isoelectric focusing for probing the conformations of different polioviral empty capsid species. Naturally occurring empty capsids (called procapsids) with an isoelectric point of 6.8 were resistant to proteolytic digestion by trypsin or chymotrypsin, as were empty capsids assembled in vitro in the presence of a cytoplasmic extract prepared from poliovirus-infected HeLa cells. In contrast, self-assembled empty capsids (isoelectric point, 5.0) were sensitive to both proteinases. Capsid proteins VP0 and VP1 were attacked predominantly, whereas VP3 was resistant to cleavage. Unpolymerized 14S particles possessed a trypsin sensitivity which was qualitatively similar to that of self-assembled empty shells. Surface-specific iodination of virions and procapsids labeled VP1 exclusively. In contrast, radioiodination of self-assembled empty capsids labeled predominantly VP0. After radioiodination the sedimentation coefficient corrected to water at 20 degrees C, the isoelectric point, and the trypsin resistance of the procapsids remained unchanged. Procapsids and extract-assembled empty capsids were N antigenic, whereas self-assembled empty capsids were H antigenic. Self assembled empty capsids were not converted to pH 6.8 trypsin-resistant structures by incubation with a virus-infected cytoplasmic extract. However, 14S particles assembled in the presence of a mock-infected extract formed empty capsids, 20% of which resembled extract-assembled empty shells as determined by the above described criteria. These and related findings are discussed in terms of empty capsid structure and morphogenesis. PMID- 6284970 TI - Regulation of protein synthesis in vesicular stomatitis virus-infected mouse L 929 cells by decreased protein synthesis initiation factor 2 activity. AB - Infection of mouse L-cell spinner cultures by vesicular stomatitis virus (VSV) effected the selective translation of viral mRNA by 4h after viral adsorption. Cell-free systems prepared from mock- and VSV-infected cells reflected this phenomenon; protein synthesis was reduced in the virus-infected cell lysate by approximately 75% compared with the mock-infected (control) lysate. This effect appeared to be specific to protein synthesis initiation since (i) methionine incorporation into protein from an exogenous preparation of initiator methionyl tRNA gave completely analogous results and (ii) the addition of a ribosomal salt wash (containing protein synthesis initiation factors) stimulated protein synthesis by the infected cell lysate but had no effect on protein synthesis by the control. Micrococcal nuclease-treated (initiation-dependent) VSV-infected cell lysates were not able to translate L-cell mRNA unless they were supplemented with a ribosomal salt wash; a salt wash from ribosomes from uninfected cells effected a quicker recovery than a salt wash from ribosomes from infected cells. When salt wash preparations from ribosomes from uninfected and infected cells were tested for initiation factor 2 (eIF-2)-dependent ternary complex capacity with added GTP and initiator methionyl-tRNA, we found that the two preparations contained equivalent levels of eIF-2. However, initiation complex formation by the factor from virus-infected cells proceeded at a reduced initial rate compared with the control. When the lysates were supplemented with a partially purified eIF-2 preparation, recovery of activity by the infected cell lysate was observed. Mechanisms by which downward regulation of eIF-2 activity might direct the selective translation of viral mRNA in VSV-infected cells are proposed. PMID- 6284972 TI - DNA sequence relationship of the baboon endogenous virus genome to the genomes of other type C and type D retroviruses. AB - Baboon endogenous virus (BaEV) is a type C retrovirus present in multiple proviral copies in the DNA of baboons. Although interspecies antigenic determinants present on reverse transcriptase and gag proteins are shared among all mammalian type C viruses, no nucleic acid homology between BaEV and other type C viruses (except RD-114) has been found in conventional liquid hybridization experiments. In this study, we used restriction fragments of cloned BaEV DNA immobilized on nitrocellulose to test for relatedness with [(32)P]cDNA's of various type C and type D viruses. We detected the following distant relationships previously found only through immunological and protein sequencing techniques: (i) eight type C viral cDNA's (the endogenous virus of rhesus monkeys, feline leukemia virus, simian sarcoma virus, gibbon ape leukemia virus, Rauscher murine leukemia virus, BALB-2, NZB, and RD-114) and two type D viral cDNA's (Mason-Pfizer monkey virus and squirrel monkey retrovirus) were able to hybridize with cloned BaEV DNA; (ii) the eight type C probes hybridized to restriction fragments spanning most of the BaEV genome, but only RD-114 hybridized to fragments within the 1.9 kilobases at the 3' end of the genome; (iii) the two type D probes hybridized primarily to fragments within the 1.9 kilobases at the 3' terminus and weakly or not at all elsewhere; and (iv) [(32)P]cDNA's of several other oncornaviruses (mouse mammary tumor virus, equine infectious anemia virus, bovine leukemia virus, and reticuloendotheliosis virus) exhibited no homology with BaEV DNA. DNA sequence analysis has allowed us to orient the BaEV restriction map with the genetic map at both ends of the genome. Homologies between retroviral cDNA's and BaEV clone restriction fragments could thus be related to specific BaEV genes. Whereas type C cDNA's hybridized to fragments from gag, pol, and the pol-env junction, squirrel monkey retrovirus cDNA hybridized only to a fragment coding for the p15E portion of env. Mason Pfizer monkey virus cDNA also hybridized within the p15E region, but exhibited homology to the 3' half of gp70 as well. These results are discussed relative to previously reported antigenic relatedness of retroviral proteins. The data suggest that BaEV represents an important link in oncornavirus evolution. PMID- 6284973 TI - Synthesis of vesicular stomatitis virus negative-strand RNA in vitro: dependence on viral protein synthesis. AB - An in vitro system is described which supports the synthesis of vesicular stomatitis virus (VSV) negative-strand RNA. The major components of this system are (i) an mRNA-dependent rabbit reticulocyte lysate to carry out cell-free protein synthesis, (ii) the five VSV mRNAs to program VSV-specific protein synthesis, and (iii) nucleocapsids containing positive- and negative-strand genome-length RNA. The protein products synthesized in the system in response to addition of saturating amounts of the five VSV mRNA's included polypeptides which comigrated in acrylamide gels with the five VSV proteins. Approximately 200 pmol of protein per ml was synthesized during a 90-min reaction. The RNA products synthesized in the system included all five of the VSV mRNA's and, in addition, negative-strand, genome-sense RNA. All of the negative-strand RNA, which represented 2 to 5% of the total RNA product synthesized in vitro, banded in CsCl at the position of nucleocapsids. All of the mature mRNA's made in the system pelleted in CsCl. This technique allowed a clear separation of negative-strand product from the mRNA products and facilitated further analysis of the negative strand product. The amount of negative-strand product produced in the system was shown to be a function of the amount of concurrent protein synthesis in the system. An increase in the level of protein synthesis led to an increase in the amount of negative-strand RNA synthesized, whereas inhibition of protein synthesis by cycloheximide resulted in a 70% inhibition of negative-strand synthesis. In contrast to the negative-strand RNA product, the amount of transcriptive product was decreased by 50% in the presence of maximum levels of viral protein synthesis. This inhibition was reversed by adding cycloheximide. Characterization of the negative-strand product by Northern blot analysis demonstrated that negative-strand product was being synthesized which hybridized to all five of the VSV mRNA's and, hence, that product representing all of the VSV cistrons was being made. This in vitro system offers an opportunity to study factors involved in the promotion of VSV genome replication as well as those responsible for the regulation of transcription. PMID- 6284974 TI - Genetic structure and transforming sequence of avian sarcoma virus UR2. AB - We have recently shown that a newly isolated avian sarcoma virus, UR2, is defective in replication and contains no sequences homologous to the src gene of Rous sarcoma virus. In this study, we analyzed the genetic structure and transforming sequence of UR2 by oligonucleotide fingerprinting. The sizes of the genomic RNAs of UR2 and its associated helper virus, UR2AV, were determined to be 24S and 35S, respectively, by sucrose gradient sedimentation. The molecular weight of the 24S UR2 genomic RNA was estimated to be 1.1 x 10(6), corresponding to 3,300 nucleotides, by gel electrophoresis under the native and denatured conditions. RNase T1 oligonucleotide mapping indicated that UR2 RNA contains seven unique oligonucleotides in the middle of the genome and shares eight 5'- and six 3'-terminal oligonucleotides with UR2AV RNA. From these data, we estimated that UR2 RNA contains a unique sequence of about 12 kilobases in the middle of the genome, and contains 1.4 and 0.7 kilobases of sequences shared with UR2AV RNA at the 5' and 3' ends, respectively. Partial sequence analysis of the UR2-specific oligonucleotides by RNase A digestion revealed that there are no homologous counterparts to these oligonucleotides in the RNAs of other avian sarcoma and acute leukemia viruses studied to date. UR2-transformed non-virus producing cells contain a single 24S viral RNA which is most likely the message coding for the transforming protein of UR2. On the basis of the uniqueness of the transforming sequence, we concluded that UR2 is a new member of the defective avian sarcoma viruses. PMID- 6284975 TI - Colobus type C virus: molecular cloning of unintegrated viral DNA and characterization of the endogenous viral genomes of Colobus. AB - The unintegrated viral DNA intermediates of colobus type C virus (CPC-1) were isolated from infected human cells that were permissive for viral growth. There were two major species of DNA, linear molecules with two copies of the long terminal repeat and relaxed circles containing only a single long terminal repeat. In addition, there was a minor species (approximately 10%) composed of relaxed circles with two copies of the long terminal repeat. A restriction endonuclease map of the unintegrated DNA was constructed. The three EcoRI fragments of circular CPC-1 DNA were cloned in the EcoRI site of lambda gtWES . lambda B and then subcloned in the EcoRI site of pBR322. Using these subgenomic fragments as probes, we have characterized the endogenous viral sequences found in colobus cellular DNA. They are not organized in tandem arrays, as is the case in some other gene families. The majority of sequences detected in cellular DNA have the same map as the CPC-1 unintegrated DNA at 17 of 18 restriction endonuclease sites. There are, however, other sequences that are present in multiple copies and do not correspond to the CPC-1 map. They do not contain CPC-1 sequences either in an altered form or fused to common nonviral sequences. Instead, they appear to be derived from a distinct family of sequences that is substantially diverged from the CPC-1 family. This second family of sequences, CPC-2, is also different from the sequences related to baboon endogenous type C virus that forms a third family of virus-related sequences in the colobus genome. PMID- 6284976 TI - Restriction endonuclease mapping of the proviral DNA of the exogenous RIII murine mammary tumor virus. AB - Cellular DNA containing integrated murine mammary tumor virus (MuMTV) was isolated from FeI/C6 feline kidney cells and CCL64 mink lung cells infected with milkborne RIII MuMTV. By using restriction enzyme HpaI, intact RIII MuMTV provirus (length, 8.7 kilobases [kb]) was excised from the cellular DNA. Subsequent restriction endonuclease analysis of this HpaI fragment with KpnI, HindIII, EcoRI, BamHI, BglII, PstI, SstI, SalI, and XhoI enabled us to construct a map of the RIII virus genome. A comparison of this map with the maps of the GR and C3H MuMTV's revealed that there are greater sequence differences between the RIII virus and the GR and C3H MuMTV proviruses than there are between the GR and C3H proviruses. The following are features of the restriction map unique to the RIII provirus: the presence of three BamHI and two EcoRI cleavage sites, a HpaI cleavage site in the terminal 3'-5' repeat unit of the provirus, and the absence of an XhoI cleavage site. Another distinguishing feature of the RIII provirus is that the sizes of some of the restriction fragments produced by cleavage of the RIII provirus with PstI are different from the sizes of the fragments obtained by PstI cleavage of the GR and C3H proviruses. Like the GR proviral DNA, the RIII proviral DNA has three SstI (SacI) cleavage sites, whereas the C3H provirus has only two SstI sites. HpaI digestion of MuMTV-infected mink lung cell DNA revealed only one class of provirus (an 8.7-kb fragment); however, we observed several minor classes of RIII proviral DNA in addition to the major class of provirus DNA in infected cat kidney cells. PstI digestion of the HpaI 8.7-kb fragments from both feline and mink cells generated a 3.7-kb DNA fragment identical in size to a PstI-generated fragment that has been found in GR and C3H milkborne virus infected cells. Although a fragment similar in size to the milkborne 3.7-kb PstI fragment has been found as an endogenous component in many C3H and GR mouse tissues, we did not observe such an endogenous fragment in the RIII mouse strain. Therefore, the 3.7-kb fragment may be useful as a marker for the milkborne RIII MuMTV provirus in RIII mice. PMID- 6284977 TI - Effect of deletions in adenovirus early region 1 genes upon replication of adeno associated virus. AB - The growth of adeno-associated virus (AAV) is dependent upon helper functions provided by adenovirus. We investigated the role of adenovirus early gene region 1 in the AAV helper function by using six adenovirus type 5 (Ad5) host range mutants having deletions in early region 1. These mutants do not grow in human KB cells but are complemented by and grow in a line of adenovirus-transformed human embryonic kidney cells (293 cells); 293 cells contain and express the Ad5 early region 1 genes. Mutants having extensive deletions of adenovirus early region 1a (dl312) or regions 1a and 1b (dl313) helped AAV as efficiently as wild-type adenovirus in 293 cells, but neither mutant helped in KB cells. No AAV DNA, RNA, or protein synthesis was detected in KB cells in the presence of the mutant adenoviruses. Quantitative blotting experiments showed that at 20 h after infection with AAV and either dl312 or dl313 there was less than one AAV genome per cell. In KB cells infected with AAV alone, the unreplicated AAV genomes were detected readily. Apparently, infection with adenovirus mutant dl312 or dl313 results in degradation of most of the infecting AAV genomes. We suggest that at least an adenovirus region 1b product (and perhaps a region 1a product also) is required for AAV DNA replication. This putative region 1b function appears to protect AAV DNA from degradation by an adenovirus-induced DNase. We also tested additional Ad5 mutants (dl311, dl314, sub315, and sub316). All of these mutants were inefficient helpers, and they showed varying degrees of multiplicity leakiness. dl312 and dl313 complemented each other for the AAV helper function, and each was complemented by Ad5ts125 at the nonpermissive temperature. The defect in region 1 mutants for AAV helper function acts at a different stage of the AAV growth cycle than the defect in the region 2 mutant ts125. PMID- 6284978 TI - Distribution of replicating simian virus 40 DNA in intact cells and its maturation in isolated nuclei. AB - The maturation of replicating simian virus 40 (SV40) chromosomes into superhelical viral DNA monomers [SV40(I) DNA] was analyzed in both intact cells and isolated nuclei to investigate further the role of soluble cytosol factors in subcellular systems. Replicating intermediates [SV40(RI) DNA] were purified to avoid contamination by molecules broken at their replication forks, and the distribution of SV40(RI) DNA as a function of its extent of replication was analyzed by gel electrophoresis and electron microscopy. With virus-infected CV-1 cells, SV40(RI) DNA accumulated only when replication was 85 to 95% completed. These molecules [SV40(RI(*)) DNA] were two to three times more prevalent than an equivalent sample of early replicating DNA, consistent with a rate-limiting step in the separation of sibling chromosomes. Nuclei isolated from infected cells permitted normal maturation of SV40(RI) DNA into SV40(I) DNA when the preparation was supplemented with cytosol. However, in the absence of cytosol, the extent of DNA synthesis was diminished three- to fivefold (regardless of the addition of ribonucleotide triphosphates), with little change in the rate of synthesis during the first minute; also, the joining of Okazaki fragments to long nascent DNA was inhibited, and SV40(I) DNA was not formed. The fraction of short-nascent DNA chains that may have resulted from dUTP incorporation was insignificant in nuclei with or without cytosol. Pulse-chase experiments revealed that joining, but not initiation, of Okazaki fragments required cytosol. Cessation of DNA synthesis in nuclei without cytosol could be explained by an increased probability for cleavage of replication forks. These broken molecules masqueraded during gel electrophoresis of replicating DNA as a peak of 80% completed SV40(RI) DNA. Failure to convert SV40(RI(*)) DNA into SV40(I) DNA under these conditions could be explained by the requirement for cytosol to complete the gap-filling step in Okazaki fragment metabolism: circular monomers with their nascent DNA strands interrupted in the termination region [SV40(II(*)) DNA] accumulated with unjoined Okazaki fragments. Thus, separation of sibling chromosomes still occurred, but gaps remained in the terminal portions of their daughter DNA strands. These and other data support a central role for SV40(RI(*)) and SV40(II(*)) DNAs in the completion of viral DNA replication. PMID- 6284979 TI - Induction of a new ribonucleotide reductase after infection of mouse L cells with pseudorabies virus. AB - The mammalian ribonucleotide reductase consists of two nonidentical subunits, protein M1 and M2. M1 binds nucleoside triphosphate allosteric effectors, whereas M2 contains a tyrosine free radical essential for activity. The activity of ribonucleotide reductase increased 10-fold in extracts of mouse L cells 6 h after infection with pseudorabies virus. The new activity was not influenced by antibodies against subunit M1 of calf thymus ribonucleotide reductase, whereas the reductase activity in uninfected cells was completely neutralized. Furthermore, packed infected cells (but not mock-infected cells) showed an electron paramagnetic resonance spectrum of the tyrosine free radical of subunit M2 of the cellular ribonucleotide reductase. These data given conclusive evidence that on infection, herpesvirus induces a new or modified ribonucleotide reductase. The virus-induced enzyme showed the same sensitivity to inhibition by hydroxyurea as the cellular reductase. The allosteric regulation of the virus enzyme was completely different from the regulation of the cellular reductase. Thus, CDP reduction catalyzed by the virus enzyme showed no requirement for ATP as a positive effector, and no feedback inhibition was observed by dTTP or dATP. The virus reductase did not even bind to a dATP-Sepharose column which bound the cellular enzyme with high affinity. PMID- 6284980 TI - Mobility of endogenous ecotropic murine leukemia viral genomes within mouse chromosomal DNA and integration of a mink cell focus-forming virus-type recombinant provirus in the germ line. AB - Characterization of endogenous ecotropic Akv proviruses in DNA of low and high leukemic mouse strains revealed the presence of one to six copies of the Akv genome per haploid genome equivalent integrated in the germ line. Low leukemic strains analyzed so far contained only one complete copy of the Akv proviral DNA. The site of integration varied among strains, although genetically related strains often carried the Akv proviral gene in the same chromosomal site. The different substrains of the AKR mouse displayed the presence of variable numbers (two to six) of Akv genomes. In all substrains one Akv genome was present in an identical chromosomal site; this locus probably comprised the progenitor genome. Closely related substrains had several Akv proviral DNAs integrated in common sites. The accumulation of Akv genomes in the germ line of the AKR/FuRdA strain is likely the result of independent integration events, since backcross studies with the Akv-negative 129 strain showed random segregation of all six proviral loci. The AKR/Cnb strain carried a recombinant provirus in the germ line. This provirus resembled in structure the AKR mink cell focus-forming viruses, which are generated by somatic recombination during leukemogenesis. Therefore, the germ line amplification of Akv proviral DNAs occurs most likely through infection of embryonic cells by circulating virus. PMID- 6284981 TI - Mutations in the herpes simplex virus DNA polymerase gene can confer resistance to 9-beta-D-arabinofuranosyladenine. AB - Mutants of herpes simplex virus type 1 resistant to the antiviral drug 9-beta-D arabinofuranosyladenine (araA) have been isolated and characterized. AraA resistant mutants can be isolated readily and appear at an appreciable frequency in low-passage stocks of wild-type virus. Of 13 newly isolated mutants, at least 11 were also resistant to phosphonoacetic acid (PAA). Of four previously described PAA-resistant mutants, two exhibited substantial araA resistance. The araA resistance phenotype of one of these mutants, PAAr5, has been mapped to the HpaI-B fragment of herpes simplex virus DNA by marker transfer, and araA resistance behaved in marker transfer experiments as if it were closely linked to PAA resistance, a recognized marker for the viral DNA polymerase locus. PAAr5 induced viral DNA polymerase activity which was much less susceptible to inhibition by the triphosphate derivative of araA than was wild-type DNA polymerase. These genetic and biochemical data indicate that the herpes simplex virus DNA polymerase gene is a locus which, when mutated, can confer resistance to araA and thus that the herpes simplex virus DNA polymerase is a target for this antiviral drug. PMID- 6284982 TI - DNA of herpesvirus pan, a third member of the Epstein-Barr virus-Herpesvirus papio group. AB - The DNA of herpesvirus pan, a primate B-lymphotropic herpesvirus, shares about 40% well-conserved sequence relatedness with Epstein-Barr virus (EBV) and herpesvirus papio DNAs. Labeled cloned fragments from the EBV recombinant DNA library were cross hybridized to blots of EcoRI, XbaI, and BamHI restriction endonuclease fragments of herpesvirus pan DNA to identify and map homologous sequences in the herpesvirus pan genome. Regions of colinear homology were demonstrated between 6 x 10(6) daltons and 108 x 10(6) daltons in the DNAs. The structural organization of herpesvirus pan DNA was similar to the format of Epstein-Barr virus and herpesvirus papio DNAs. The DNA consists of two domains of largely unique sequence complexity, a segment US of 9 x 10(6) daltons and a segment UL of 88 x 10(6) daltons. US and UL are separated by a variable number of tandem repetitions of a sequence IR (2 x 10(6) daltons). There was homology between DNA which mapped at 26 to 28 x 10(6) daltons and 93 to 95 x 10(6) daltons in UL. The terminal reiteration component, TR, of herpesvirus pan DNA and sequences which mapped to the left of 6 x 10(6) daltons and to the right of 108 x 10(6) daltons had no detectable homology with the corresponding regions of Epstein-Barr virus DNA. PMID- 6284983 TI - Maturation of the envelope glycoproteins of Newcastle disease virus on cellular membranes. AB - Based on subcellular fractionation data, the following maturation pathways were proposed for the Newcastle disease virus glycoproteins. During or shortly after synthesis in rough endoplasmic reticulum, hemagglutinin-neuraminidase (HN) and fusion (F0) glycoproteins underwent dolichol pyrophosphate-mediated glycosylation, and HN assumed a partially trypsin-resistant conformation. HN began to associate into disulfide-linked dimers in rough endoplasmic reticulum, and at least one of its oligosaccharide side chains was processed to a complex form en route to the cell surface. During migration in intracellular membranes, F0 was proteolytically cleaved to F1.2. Neither HN nor F1,2 required oligosaccharide side chains for migration to plasma membranes, and cleavage of F0 also occurred without glycosylation. Virion- and plasma membrane-associated HN contained both complex and high-mannose oligosaccharide chains on the same molecule, and F1,2 contained at least high-mannose forms. Several of the properties of HN were notable for a viral glycoprotein. The oligosaccharide side chains of HN were modified very slowly in chick cells, whereas those of the G glycoprotein of vesicular stomatitis virus were rapidly processed to a complex form. Therefore, their different rates of migration and carbohydrate processing were intrinsic properties of these glycoproteins. Consistent with its slow maturation, the HN glycopolypeptide accumulated to high levels in intracellular membranes as well as in plasma membranes. Intracellular HN contained immature oligosaccharide side chains, suggesting that it accumulated in the pre Golgi/Golgi segment of the maturation pathway. The major site of accumulation of mature HN with neuraminidase activity was the plasma membrane. PMID- 6284984 TI - Adsorption of Rous sarcoma virus to genetically susceptible and resistant chicken cells studied by laser flow cytometry. AB - Quantitative binding of Rous sarcoma virus (RSV) of different antigenic subgroups to chicken cells was examined by using a laser flow cytometer/cell sorter. RSV of subgroups A, C, and E, labeled with the fluorescent membrane probe rhodamine-18, bound 2 to 10 times more to genetically susceptible chicken embryo fibroblasts than to resistant cells, as measured by flow cytometry on a single-cell basis. This suggested that susceptible cells possess both specific and nonspecific receptors for virus adsorption, whereas resistant cells bind virus only by means of nonspecific sites. Polybrene at low concentration increased eightfold the binding of virus. Higher levels of Polybrene inhibited adsorption. Cell binding sites were saturable, and attachment of labeled virus could be partially blocked by preexposure of cells to unlabeled RSV. Virus surface glycoproteins played an important role in adsorption, since their removal with bromelain decreased binding of virus to susceptible cells. Maximal binding of RSV to both susceptible and resistant cells occurred within 10 min, although the level of binding was up to 10-fold higher for susceptible cells. Binding to all cell types showed a broad distribution. This implies that there are considerable differences in the number of virions bound per cell. PMID- 6284985 TI - DNA sequence of the 5' terminus containing the replication origin of parvovirus replicative form DNA. AB - The nucleotide sequence of the 5' terminus of the parvovirus H-1 was determined. There are two orientations of the 242-base-pair terminal palindrome in native replicative form DNA, one inverted with respect to the other. Adjacent to the terminal palindrome is an AT-rich region that is noncoding and contains a 55-base pair tandem repeat. The addition mutant of H-1, DI-1, was also sequenced in this region and shown to have three copies of the tandem repeat sequence. Similarly, the related parvovirus H-3 contains only one copy of this repeat sequence. This region contains the replication origin for parvovirus replicative form DNA replication. Some of the implications of these results are discussed. PMID- 6284986 TI - Molecular cloning of the Fujinami sarcoma virus genome and its comparison with sequences of other related transforming viruses. AB - Full-length proviral DNA of Fujinami sarcoma virus (FSV) of chickens was molecularly cloned and characterized. An analysis of FSV DNA integrated in mammalian cells showed that restriction endonuclease SacI has a single cleavage site on FSV DNA. Unintegrated closed circular FSV DNA obtained from newly infected cells was linearized by digestion with SacI and cloned into lambdagtWES.lambdaB. The following three different molecules were isolated: FSV-1 (4.4 kilobases [kb]) and FSV-2 (4.7 kb), which appeared to be full-length FSV DNA molecules containing either one or two copies of the long terminal repeat structure, and FSV-3 (6 kb), which consisted of part FSV DNA and part DNA of unknown origin. An analysis of the structure of cloned FSV-1 and FSV-2 DNA molecules by restriction endonuclease mapping and hybridization with appropriate probes showed that about 2.6 kb of the FSV-unique sequence called FSV-fps is located in the middle of the FSV genome and is flanked by helper virus-derived sequences of about 1.3 kb at the 5' end and 0.5 kb at the 3' end. The long terminal repeats of FSV were found to have no cleavage site for either EcoRI or PvuI. Upon transfection, both FSV-1 DNA and FSV-2 DNA were able to transform mammalian fibroblasts. Four (32)P-labeled DNA fragments derived from different portions of the FSV-fps sequence were used for hybridization to viral RNAs. We found that sequences within the 3' half of the FSV-fps gene are homologous to RNAs of PRCII avian sarcoma virus and the Snyder-Theilen strain of feline sarcoma virus, both of which were previously shown to contain transforming genes related to FSV-fps. These results suggest that the 3' portion of the FSV-fps sequence may be crucial for the transforming activity of fps-related oncogenic sequences. PMID- 6284987 TI - Identification of the initiation site of poliovirus polyprotein synthesis. AB - The complete nucleotide sequence of poliovirus RNA has a long open reading frame capable of encoding the precursor polyprotein NCVP00. The first AUG codon in this reading frame is located 743 nucleotides from the 5' end of the RNA and is preceded by eight AUG codons in all three reading frames. Because all proteins that map at the amino terminus of the polyprotein (P1-1a, VP0, and VP4) are blocked at their amino termini and previous studies of ribosome binding have been inconclusive, direct identification of the initiation site of protein synthesis was difficult. We separated and identified all of the tryptic peptides of capsid protein VP4 and correlated these peptides with the amino acid sequence predicted to follow the AUG codon at nucleotide 743. Our data indicate that VP4 begins with a blocked glycine that is encoded immediately after the AUG codon at nucleotide 743. An S1 nuclease analysis of poliovirus mRNA failed to reveal a splice in the 5' region. We concluded that synthesis of the poliovirus polyprotein is initiated at nucleotide 743, the first AUG codon in the long open reading frame. PMID- 6284988 TI - Genetic analysis of murine hepatitis virus strain JHM. AB - We performed a genetic analysis of 37 temperature-sensitive mutants of murine hepatitis virus strain JHM. Of our mutants, 32 did not induce murine hepatitis virus-specific RNA synthesis in infected cells at the restrictive temperature, 39 degrees C. By complementation testing we have identified at least seven nonoverlapping complementation groups. Six of the genes identified in this way are required for murine hepatitis virus-specific RNA synthesis. The seventh complementation group is made up of five mutants which induced virus-specific RNA synthesis at 39 degrees C. PMID- 6284989 TI - Cloning of two genetically transmitted Moloney leukemia proviral genomes: correlation between biological activity of the cloned DNA and viral genome activation in the animal. AB - The Mov-7 and Mov-9 substrains of mice, carrying Moloney murine leukemia virus (M MuLV) in their germ line at the Mov-7 locus and Mov-9 locus, respectively, are different with respect to virus activation. Infectious virus appears in all mice carrying the Mov-9 locus but is not activated in animals carrying the Mov-7 locus. Consequently, only Mov-9 mice develop viremia and subsequent leukemia. The endogenous M-MuLV provirus with flanking mouse sequences corresponding to the Mov 7 and Mov-9 loci was molecularly cloned. Detailed restriction maps obtained from the cloned DNAs revealed no detectable differences in the proviral genomes. The flanking mouse sequences, however, were different, confirming that the Mov-7 and Mov-9 loci represent different integration sites of M-MuLV. Both clones induced XC plaques in a transfection assay. The specific infectivity of the clones, however, was different. A total of 10(-5) XC plaques per genome equivalent were induced by the Mov-9 clone, whereas only 10(-9) XC plaques per genome equivalent were induced by the Mov-7 clone. Moreover, NIH 3T3 cells transfected with the Mov 9 clone produced NB-tropic M-MuLV, whereas cells transfected with the Mov-7 clone did not produce infectious virus. The results suggest that M-MuLV integrated at the Mov-7 locus carries a mutation which prevents synthesis of infectious virus but permits XC plaque induction by partial genome expression or synthesis of noninfectious particles. Thus, the pattern of virus expression in Mov-7 and Mov-9 mice correlates with the biological properties of the respective clones. Genomic DNA from Mov-9 mice was not infectious in the transfection assay (specific infectivity < 10(-7) PFU per genome equivalent). As the only difference between the genomic and the cloned Mov-9 DNA appears to be the presence of 5 methylcytosine in CpG sequences, our results suggest that removal of methyl groups by molecular cloning in procaryotes permits genome expression in transfected eucaryotic cells. Our results support the hypothesis that DNA methylation is relevant not only in genome expression in the animal but also in expression of genes transfected into eucaryotic cells. PMID- 6284991 TI - Antibody to a host protein prevents initiation by the poliovirus replicase. AB - In vitro transcription of poliovirus RNA was catalyzed by the combination of a virus-coded polymerase and a host cell protein (host factor). Antibody to host factor inhibited template-dependent synthesis of complementary RNA where presumably RNA chain initiation occurred. On the contrary, elongation of already initiated RNA chains catalyzed by the replicase-template complex was not inhibited by anti-host factor antibody. These results strongly favored our previous notion that the host factor was needed for the initiation step of viral complementary RNA synthesis. PMID- 6284990 TI - Nucleotide sequence analysis of the long terminal repeat of integrated simian sarcoma virus: evolutionary relationship with other mammalian retroviral long terminal repeats. AB - Nucleotide sequence analysis of the long terminal repeat (LTR) of the integrated simian sarcoma virus showed that the simian sarcoma virus LTR comprised 504 nucleotides with an inverted repeat of seven bases at its 5' and 3' termini. At the site of simian sarcoma virus integration, cellular flanking sequences adjacent to the proviral LTR contained a direct repeat of four bases. A 13-base sequence after the 5' LTR was found to be complementary to prolyl tRNA, suggesting that tRNAPro may serve as the primer for reverse transcription of simian sarcoma virus RNA. The U5 and R regions, derived respectively from the 5' end and terminally redundant sequences of the viral RNA, were found to have similar organization and sequence homology close to that of Moloney murine sarcoma virus or Moloney murine leukemia virus. These results indicate that regions within LTRs with known functionally important sequences have been most well conserved during retrovirus evolution. PMID- 6284992 TI - Rapid method for the preparation of encephalomyocarditis virus protease from rabbit reticulocyte lysates. AB - Fractionation of the reticulocyte lysate translation products of encephalomyocarditis virus RNA by ultracentrifugation showed that the viral proteins were distributed differentially in the supernatant and the ribosomal pellet fractions. The viral noncapsid proteins C and D, which contain the viral protease sequence, sedimented preferentially with the pellet fraction. Incubation of the resuspended pellet and subsequent centrifugation of the suspension resulted in cleavage of the protease from proteins C and D and separation of the enzyme from reticulocyte particulate proteins. Preparations thus obtained contained only three encephalomyocarditis virus proteins and were almost devoid of reticulocyte proteins. PMID- 6284993 TI - Presence in growth-arrested cells of cellular proteins that interact with simian virus 40 small-t antigen. AB - Two cellular proteins, 56K and 32K, found in association with simian virus 40 small-t antigen were not induced by viral infection. In addition, the proteins were expressed by cells in the growth arrest period, a time in which small-t function is of importance in infection and transformation. PMID- 6284994 TI - Isolation and characterization of c-myc, a cellular homolog of the oncogene (v myc) of avian myelocytomatosis virus strain 29. AB - The chicken genome contains nucleotide sequences homologous to transforming genes (oncogenes) of a number of avian retroviruses. We have isolated chicken DNA (c myc) that is homologous to the oncogene (v-myc) of the avian myelocytomatosis virus MC29 and have compared the structures of the cellular and viral genes. Results from restriction endonuclease mapping of c-myc and from analysis of heteroduplexes between the DNAs of the cellular and viral genes show that c-myc is homologous to 1,500 nucleotides in v-myc DNA. This homologous region is interrupted in c-myc by an intron-like sequence of 1,100 nucleotides which is absent from v-myc. Nuclear RNA from normal chicken cells contains at least five species of transcripts from c-myc ranging from 2.5 to 6.5 kilobases in length. By contrast, cytoplasm contains only the 2.5-kilobase c-myc RNA. These features of the c-myc gene and its nuclear transcripts are characteristic of normal cellular genes and suggest that the myc gene is of cellular rather than viral origin. The exons in c-myc may define two functional domains in the gene and may therefore facilitate the dissection of the different oncogenic potentials of the MC29 virus. PMID- 6284995 TI - Role of p60src kinase activity in the induction of neuroretinal cell proliferation by rous sarcoma virus. AB - Expression of the src gene of Rous sarcoma virus (RSV) in chicken embryo neuroretinal (NR) cells results in morphological transformation and sustained proliferation of a normally resting cell population. We have previously reported the isolation of mutants of RSV which retain full growth-promoting activity while displaying reduced transforming properties. Two such mutants, PA101 and PA104, were used to investigate whether the p60src-associated kinase activity is required for the mitogenic function of src. A comparison of the patterns of phosphorylation of wild-type and mutant p60src revealed that the phosphorylation of tyrosine residues of p60src of PA104 was markedly reduced, whereas the relative amount of phosphotyrosine in p60src of PA101 was comparable to that of the wild-type protein. In vitro kinase activity of p60src immunoprecipitated from NR cells infected with PA101 or PA104 as measured by phosphorylation of the heavy chains of specific immunoglobulin G molecules was 1/10 that of the wild-type molecule. Moreover, when NR cells infected with mutants temperature sensitive for mitogenic capacity were maintained at a temperature either permissive or restrictive for cell growth, quantitation of kinase activity indicated that proliferation of NR cells could not be linked to the absolute level of in vitro kinase activity of p60src. Transformation of NR cells by wild-type RSV resulted in a 10-fold increase in total cellular phosphotyrosine and in the phosphorylation of tyrosine residues of a 34K protein, a possible in vivo substrate for p60src. In contrast, phosphorylation of tyrosine residues of cellular targets was markedly reduced in NR cells infected with PA101 or PA104. These results indicate that the mitogenic capacity of RSV in NR cells does not require elevated levels of p60src kinase activity. PMID- 6284996 TI - Evidence for simian virus 40 late transcriptional control: mixed infections of wild-type simian virus 40 and a late leader deletion mutant exhibit trans effects on late viral RNA synthesis. AB - Mixed infections involving equal multiplicities of wild-type simian virus 40 and viable deletion mutant dl861 resulted in decreased cytoplasmic levels of wild type-derived male mRNA, as well as very low to undetectable levels of mutant derived late mRNA, as compared with individual infections. The dl861 deletion removes 16 to 25 base pairs from the late leader region. This deletion was shown to be the direct cause of the mixed-infection effect; replacement of the deletion with wild-type sequences restored normal levels of late mRNAs in mixed infections. Other viral functions, e.g., early gene expression and replication, were found to be unaffected by the dl861 deletion. Further examination of the mixed-infection effect showed that the levels of unspliced nuclear precursors of late mRNA, derived from both the mutant and wild-type genomes, were decreased or undetectable, in accord with the cytoplasmic results. Thus, the effect appears to be occurring at the transcriptional level. These data demonstrate a trans-acting effect on late transcription, which is detectable due to the presence of the dl861 mutant in the mixed infection. This finding is indicative of a diffusible factor which exerts a control on simian virus 40 late gene expression at the transcriptional level. A model for positive control of simian virus 40 late gene expression is presented. PMID- 6284997 TI - Functional characterization of cleavage-defective mutants of encephalomyocarditis virus. AB - We characterized seven temperature-sensitive capsid cleavage (cleavage-defective) mutants of encephalomyocarditis virus. Our experimental approach was to monitor in vitro proteolysis reactions of either wild-type or cleavage-defective mutant capsid precursors mixed with cell-free translation products (containing the viral protease) of either wild-type or mutant viral RNA. The cell-free translation reactions and in vitro proteolysis reactions were done at 38 degrees C, because at this temperature cleavage of the capsid precursors was restricted in reactions containing cleavage-defective mutant viral RNA as the message, relative to those reactions containing wild-type viral RNA as the message. Wild-type or cleavage defective mutant capsid precursors were prepared by adding cycloheximide to cell free translation reactions primed with wild-type or mutant viral RNA, respectively, 12 min after the initiation of translation. In vitro proteolysis of wild-type capsid precursors with cell-free translation products of either wild type or cleavage-defective mutant viral RNA led to similar products at 38 degrees C, indicating that the cleavage-defective mutant viral protease was not temperature sensitive. As a corollary to this, at 38 degrees C cleavage-defective mutant capsid precursors were not cleaved as completely as were wild-type capsid precursors by products of cell-free translation of wild-type viral RNA. The results from these in vitro proteolysis experiments indicate that all seven of the cleavage-defective mutants have capsid precursors with a temperature sensitive configuration. PMID- 6284999 TI - Nucleotide sequence analysis of the long terminal repeat of avian myeloblastosis virus and adjacent host sequences. AB - The nucleotide sequence of the integrated avian myeloblastosis virus long terminal repeat has been determined. The sequence is 385 base pairs long and is present at both ends of the viral DNA. The cell-virus junctions at each end consist of a 6-base-pair direct repeat of cell DNA next to the inverted repeat of viral DNA. The long terminal repeat also contains promoter-like sequences, an mRNA capping site, and polyadenylation signals. Several features of this long terminal repeat suggest a structural and functional similarity with sequences of transposable and other genetic elements. Comparison of these sequences with long terminal repeats of other avian retroviruses indicates that there is a great variation in the 3' unique sequence (U3), whereas the 5' specific sequences (U5) and the R region are highly conserved. PMID- 6284998 TI - Identification, synthesis, and modifications of simian rotavirus SA11 polypeptides in infected cells. AB - The synthesis and processing of simian rotavirus SA11 polypeptides was investigated after infection of MA104 cells. [35S]methionine- or 3H-amino acid labeled cell extracts were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Viral protein synthesis was maximal 3 to 5 h postinfection, and 12 major viral polypeptides were detected. Immunoprecipitation and peptide mapping experiments, demonstrated five viral structural proteins (125,000 daltons [125K], 94K, 88K, 41K, and 38K). Three proteins (53K, 35K, and 34K) were identified as nonstructural by comparison of their partial proteolysis maps with those from polypeptides of similar molecular weight synthesized in vitro from viral RNA transcripts. Assignment as to structural or nonstructural status of two other primary gene products (26K and 20K) remains tentative. Pulse-chase experiments and tunicamycin blockage of glycosylation revealed cotranslational or post-translational modifications (or both) and precursor-product relationships of several of the polypeptides. Tunicamycin inhibition of glycosylation identified a 35.5K polypeptide which was proven to be the precursor to the 38K structural glycoprotein by immunoprecipitation and peptide mapping analyses. Tunicamycin treatment of infected cells also resulted in the disappearance of other glycoprotein species (23K to 29K) and in the concomitant build-up of an unglycosylated 20K polypeptide, suggesting a precursor-product relationship between those polypeptides. Labeling with [3H]glucosamine or [3H]mannose suggested that the rotavirus glycoproteins contained high mannose oligosaccharides. The effects of amino acid analogs on rotavirus polypeptide synthesis and processing were also investigated. PMID- 6285000 TI - Characterization of two RNA polymerase activities induced by mouse hepatitis virus. AB - RNA-dependent RNA polymerase activity was found in mouse hepatitis virus strain A59 (MHV-A59)-infected cells. The enzyme was induced in the infected cells and could not be detected in the MHV-A59 virion. Two peaks of RNA polymerase activity, one early and the other late in infection, were detected. These polymerase activities were in temporal sequence with early and late virus specific RNA synthesis. Both of them were found to be associated with membrane fractions. There were significant differences in the enzymatic properties of the two polymerases. The early polymerase, but not the late polymerase, could be activated by potassium ions in the absence of magnesium ions and also had a lower optimum pH than the late polymerase. It was therefore probable that the enzymes represent two different species of RNA polymerase and perform different roles in virus-specific RNA synthesis. The effects of cycloheximide on MHV-specific RNA synthesis were determined. Continuous protein synthesis was required for both early and late RNA synthesis and might also be required for shutoff of early RNA synthesis. PMID- 6285001 TI - Mutant of simian virus 40 large T-antigen that is defective for viral DNA synthesis, but competent for transformation of cultured rat cells. AB - A mutant was isolated which demonstrates that the transforming activity of simian virus 40 large T-antigen is separable from its function in viral DNA replication. The mutant, SVR9D, is nonconditionally defective for viral DNA synthesis, but competent at wild-type level for morphological transformation of cultured rat cells. The lytic growth defect in SVR9D is complemented by the simian virus 40 A gene product present in the transformed CV1 cell line, COS1. The lesion in SVR9D DNA was mapped genetically by marker rescue of plaque formation and localized to a 214-base-pair segment of the viral genome bounded by nucleotide numbers 4100 and 4314. DNA sequence analysis showed the mutation to be an adenine-to-guanine transition at nucleotide number 4178. This change predicts a lysine-to-glutamic acid amino acid change at residue number 214 of the mutant large T-antigen polypeptide. PMID- 6285002 TI - Conservation of protein coding potential in the long terminal repeats of exogenous and endogenous mouse mammary tumor viruses. AB - In vitro protein synthesis and DNA sequence analysis indicate that mouse mammary tumor virus differs from other well-characterized retroviruses in that the long terminal repeat region of the provirus has the capacity to encode proteins. Different exogenously transmitted mouse mammary tumor virus strains and endogenous proviral units conserved this open reading frame feature in the long terminal repeat despite a variation in nucleotide sequence. The proteins encoded by the different long terminal repeats were clearly related, but showed minor variations in size and tryptic peptide maps. In each case, the largest in vitro product had a molecular weight of about 36,000 to 37,000, suggesting that the open reading frame sequences must extend for approximately 1,000 nucleotides beginning at the extreme 5' end of the long terminal repeat. The fact that the reading frame was conserved among these viruses argues in favor of an in vivo function for the open reading frame protein. PMID- 6285003 TI - Identification and characterization of a group of discrete initiated oligonucleotides transcribed in vitro from the 3' terminus of the N-gene of vesicular stomatitis virus. AB - Four triphosphate-initiated oligonucleotides, 11 to 14 bases long, produced by in vitro transcription of vesicular stomatitis virus were identified as the uncapped 5' sequences of N-gene mRNA. Characterization of these oligonucleotides reveals that they are continually produced stable transcripts that do not remain template bound. Under the conditions used, the oligonucleotide transcripts were produced at 8 to 10 times the molar amount of leader, suggesting that the N-gene mRNA is internally initiated. PMID- 6285004 TI - In vitro synthesis of triphosphate-initiated N-gene mRNA oligonucleotides is regulated by the matrix protein of vesicular stomatitis virus. AB - Wild-type Indiana virus transcribed four 11- to 14-nucleotide-long, 5' N-gene mRNA sequences in vitro. The amount of oligonucleotides synthesized relative to leader by wild-type virions varied inversely with the salt concentration of the transcription reaction. Reduced oligonucleotide synthesis by nucleocapsids at all salt concentrations tested and a comparison of the proteins remaining bound to the template of nucleocapsids and virions transcribed in different NaCl concentrations suggested that the matrix (M) protein regulates oligonucleotide synthesis. Examination of the transcription products synthesized in no NaCl and 0.144 M NaCl by an M-protein mutant and an increase in oligonucleotide synthesis by nucleocapsids when purified M-protein was added to transcription reactions confirmed M-protein's role in oligonucleotide synthesis. Wild-type virion mRNA synthesis was inhibited, and oligonucleotide synthesis was greater than leader synthesis at high virus concentrations. As the virus was diluted, inhibition of mRNA synthesis was relieved and oligonucleotide synthesis was reduced. The M protein mutant tsG33 exhibited neither transcription inhibition at high virus concentrations nor the reciprocal synthesis of mRNA and the oligonucleotides seen with wild-type virions. These results are entirely consistent with the stop-start model of transcription and suggest a model for the control of transcription by M protein. PMID- 6285005 TI - Accumulation of spliced avian retrovirus mRNA is inhibited in S adenosylmethionine-depleted chicken embryo fibroblasts. AB - The synthesis and processing of B77 avian sarcoma virus RNA in infected chicken embryo fibroblasts was followed in the presence and absence of cycloleucine, a competitive inhibitor of the synthesis of S-adenosylmethionine and thus an inhibitor of RNA methylations. An increase in the steady-state levels of genome length RNA and a decrease in the steady-state levels of subgenomic RNA molecules were obtained in the S-adenosylmethionine-depleted avian sarcoma virus-infected cells after 24 h of treatment with the inhibitor. The total number of virus specific RNA molecules per cell, however, remained relatively constant under either condition. The production of newly synthesized virus-specific RNA in cycloleucine-treated and untreated cells infected with a transformation-defective strain of B77 avian sarcoma virus was followed as a function of [(3)H]uridine labeling time. The accumulation of radioactive genome-length 8.4-kilobase (kb) RNA continued in cycloleucine-treated cells, and virus particle production proceeded at normal rates as previously shown by incorporation of labeled nucleoside precursors or amino acids. In contrast, newly synthesized 3.5-kb subgenomic mRNA, the putative mRNA for the envelope protein precursor, failed to accumulate in the treated cells. The extent of the inhibition in the appearance of the radioactive 3.5-kb RNA was correlated with the extent of the inhibition of viral genomic and cellular mRNA methylations and was a function of the cycloleucine concentration. Under conditions in which the accumulation of 3.5-kb envelope protein mRNA was blocked by the cycloleucine treatment, there were significant increases in the rate of synthesis of the polypeptide products of the genome-length RNA, the precursors to the non-glycosylated gag proteins (Pr76(gag)), and the reverse transcriptase (Pr 180(gag pol)) relative to the rate of synthesis of the envelope protein precursor (gPr 92(env)). These results suggest that there is an S-adenosylmethionine requirement for the splicing, but not for the synthesis, packaging, or messenger function, of avian retrovirus genome-length RNA. Possible reasons for this requirement are discussed. PMID- 6285006 TI - Evidence for an interaction between the membrane protein of a paramyxovirus and actin. AB - Evidence for an interaction of the membrane (M) protein of Newcastle disease and Sendai viruses with cellular actin was obtained by three different techniques. M protein linked to Sepharose 4B was found to bind actin, but not myoglobin or bovine serum albumin, and to selectively remove actin from a mixture of these three proteins. Sedimentation of a mixture of M protein and F-actin through a sucrose gradient resulted in sedimentation of M protein with actin. Control proteins, bovine serum albumin and cytochrome c, did not sediment with actin. In circular dichroism studies, M protein added to actin in a 1:1 complex resulted in a significant increase in negative ellipticity at 220 nm, which corresponds to an increase in alpha-helix and a decrease in beta-structure and random coil. This is indicative of an interaction between M protein and actin. It is possible that the frequent identification of cellular actin in a number of enveloped viruses may be attributed to the interaction of actin and M protein or its equivalent. PMID- 6285008 TI - Pseudorabies virus infection in raccoons: a review. AB - Pseudorabies is a rarely reported disease of raccoons. Laboratory and field evidence of PRV infection suggests the raccoon is a "dead end" host with little opportunity for raccoon-to-raccoon spread of virus. All reported field cases have been associated closely with infected swine and swine have been considered the source of the raccoon infection. The clinical signs of PRV in raccoons closely resembles those of canine distemper and rabies virus infections. Infection with the latter viruses are considered more prevalent and likely to be mistaken for PRV infection. Both CD and rabies virus may be maintained in raccoon populations with raccoon-to-raccoon transfer while PRV may not. Differentiation of PRV, CD and rabies infections is best achieved by histopathologic analysis of lung and brain tissue, together with virus isolation. It is of utmost public health importance that wildlife authorities recognize the similarities between these diseases, together with the different epidemiologic behavior of the viruses and the means to differentiate clinical cases. PMID- 6285009 TI - Avian pox in birds from Trinidad. AB - Over a 7-year period in Trinidad, 9,514 birds were examined for avian pox and four species were found infected: the golden-headed manakin, Pipra erythrocephala (7% infected), the white-bearded manakin, Manacus manacus (5%), the violaceous euphonia Euphonia violacea (1%), and the bare-eyed thrush, Turdus nudigenis (less than 1%). The elaborate courtship displays of manakins may have a bearing on a "common source" type of infection. The apparently abrupt appearance of the disease at three localities in Trinidad in 1964 perhaps indicates introduction of the virus by migratory birds. PMID- 6285007 TI - Change of DNA near the origin of replication enhances the transforming capacity of human papovavirus BK. AB - A turbid-plaque-forming mutant (pm522) of human papovavirus BK, which has a small deletion at about 0.7 map unit and grows somewhat more slowly in human cells than does wild-type BK virus, transformed hamster and rat cells in culture much more efficiently than did wild-type virus. Another plaque morphology mutant, pm525, forming turbid plaques larger than those of pm522 also had a high transforming capacity. The similar difference in transforming capability between wild-type and plaque morphology viruses was observed with DNAs extracted from virions. Recombinant viruses were constructed from the wild-type DNA fragment lacking HindIII-C (0.62 to 0.73 map unit) and pm522 HindIII-C (including the origin of replication) by the molecular cloning method. Characterization of the recombinants showed that the change near the origin of DNA replication was responsible both for the altered plaque morphology and for the enhanced transforming capacity of the BK virus mutant. PMID- 6285010 TI - Prevalence of antibodies of selected infectious disease agents in the peninsular desert bighorn sheep (Ovis canadensis cremnobates) of the Santa Rosa Mountains, California. PMID- 6285011 TI - Pretreatment peripheral blood counts in small-cell carcinoma of the lung. PMID- 6285012 TI - Dilatation and evacuation procedures and second-trimester abortions. The role of physician skill and hospital setting. AB - Some clinicians have hesitated to perform dilatation and evacuation (D & E) procedures at 13 weeks' gestation or later because D & Es are more difficult to perform safely than suction-curettage procedures. Moreover, many clinicians still believe all second-trimester abortion procedures should be performed in a hospital. To evaluate these concerns, we analyzed 24,664 abortion performed between 1973 and 1978 by four physicians associated with a large outpatient abortion facility; 3,711 (15%) of the abortions were second-trimester procedures. Dilatation and evacuation was associated with a lower rate of serious complications per 100 procedures (0.23) than instillation of either dinoprost (prostaglandin F2 alpha) (1.28) or hypertonic saline (2.26). In addition, D & E had lower rates for most other specific complications. We conclude that D & E, while requiring more operator skill than earlier suction-curettage procedures, can be learned by gynecologists familiar with suction-curettage, can be performed more safely than the alternative instillation procedures, and can be safely practiced in selected ambulatory settings. PMID- 6285013 TI - Successful treatment of adrenal metastases from large-cell carcinoma of the lung. AB - Although on a weight basis the adrenal gland is the most common site of extranodal spread from lung cancer, antemortem diagnosis and successful therapy of adrenal metastasis are rare. We have treated two patients with apparently solitary metastases in the adrenal gland from large-cell carcinomas of the lung using a combination of adrenalectomy and vigorous therapy to the primary site. Both patients experienced symptomatic relief, and, in contrast to the expected short survival in lung cancer with distant metastasis, they remain well six years and 14 years after treatment. PMID- 6285014 TI - Antiandrogen effects of ulcer drug averted with experimental agent. PMID- 6285015 TI - [Clinical studies on 9, 3"-diacetylmidecamycin in respiratory tract infections in the field of pediatrics (author's transl)]. AB - Laboratory and clinical studies were performed on 9, 3"-diacetylmidecamycin (MOM), a new macrolide antibiotic in the field of pediatrics, and the results were as follows. Antibacterial activity: For 32 clinically isolated strains of Staphylococcus aureus, the MIC of MOM ranged from 0.78 to 1.56 micrograms/ml for 17 of the 32 strains, and exceeded 100 micrograms/ml for the 15 remaining strains with both inoculum sizes of 10(8) cells/ml and 10(6) cells/ml. For 27 strains of Streptococcus pyogenes, the MIC range was wide, varying from 0.10 to greater than or equal to 100 micrograms/ml and less than 1.56 micrograms/ml for about 2/3 of all the 27 strains. For 9 strains of Bordetella pertussis, the MIC ranged from 0.10 to 0.78 microgram/ml and 0.10 to 0.39 microgram/ml with the inoculum size of 10(8) cells/ml and 10(6) cells/ml, respectively. Comparing the antibacterial activity of MOM with that of midecamycin (MDM) and erythromycin (EM) against these 3 bacterial species, MOM was almost comparable to MDM, but about 2 or 3 tubes inferior to EM. Absorption and excretion: MOM was administered to 5 children (from 5 to 8 years old) at a dose of 10 mg/kg or 20 mg/kg at 30 minutes before breakfast. The peak of serum concentration was observed 30 minutes to 1 hour after administrations of both dosages: 0.52 to 1.71 micrograms/ml with 10 mg/kg and 0.88 to 1.77 micrograms/ml with 20 mg/kg. 0.09 to 1.10% and 0.94 to 1.19% of MOM were excreted in the urine within the first 6 hours, respectively. CLINICAL RESULTS: MOM was administered to 28 pediatric patients with acute respiratory tract infections (acute pharyngitis; 2, acute purulent tonsillitis; 19, acute bronchitis; 4, acute pneumonia; 2 and whooping cough; 1). The overall clinical response was excellent in 10, good in 10, fair in 3 and poor in 5; the efficacy rate was 71.4%. Isolated S. pyogenes strains were eradicated in 6 out of 11 strains, reduced in 3 and unchanged in 2 strains. One strain of S. aureus was eradicated. One strain of non group A beta-Streptococcus was reduced. Haemophilus influenzae strains were reduced in 1 of the 4 strains and unchanged in 3 strains. The overall eradication rate was 41.2%. No side effects or abnormal laboratory findings were observed, but 1 case complained of a bitter taste. PMID- 6285016 TI - [Basic studies of cefotiam (author's transl)]. AB - The basic studies of cefotiam (CTM) were performed, and the following results were obtained. 1. One shot intravenous injection of 40 mg/kg of CTM (1) Serum levels: The mean levels of CTM in 3 children were 133.9 micrograms/ml after 15 minutes, 71.5 micrograms/ml after 30 minutes 32.6 micrograms/ml after 1 hour, 9.2 micrograms/ml after 2 hours, 2.5 micrograms/ml after 4 hours, 0.9 microgram/ml after 6 hours. The serum half life of CTM was approximately 1.2 hours on beta phase. (2) Urinary excretion: The mean urinary excretions were 39.4% over 3 hours, 44.7% over 4 hours, 45.7% over 6 hours. 2. 1 hour-drip infusion of 40 mg/kg of CTM (1) Serum levels: The mean serum levels of CTM in 4 children were 140.1 micrograms/ml at immediately after drip infusion was completed, 10.6 micrograms/ml after 1 hour, 3.8 micrograms/ml after 2 hours, 1.4 micrograms/ml after 3 hours, 0.5 microgram/ml after 5 hours. The serum half life of CTM was approximately 1.0 hour on beta-phase. (2) Urinary excretions: The mean urinary excretions were 42.9% over 2 hours, 48.0% over 4 hours, 48.7% over 6 hours. 3. Bioactive metabolite in serum and urine: No bioactive metabolites were observed in either serum or urine after administration of 40 mg/kg of CTM. PMID- 6285017 TI - [Sensitivity to second-generation cephem agents of clinically isolated strains of bacteria in otorhinolaryngological field (author's transl)]. AB - Clinically isolated 741 strains of 10 bacterial genus (except for Pseudomonas aeruginosa) which are frequent causal pathogens of infection in the otorhinolaryngological field were examined for their sensitivity to second generation cephem agents, CTM, CXM and CMZ using the first-generation agent, CEZ, as a control. 1) The antibacterial activity of CTM, CXM, and CMZ was expanded to H. influenzae and Proteus spp. (indole positive) as compared with the first generation agent, CEZ. CTM and CXM and strong antibacterial activity against H. influenzae. CMZ was slightly inferior to them in this activity. In contrast, against Proteus spp. (indole positive) CMZ was most effective and CTM was more effective than CXM. 2) The proportion of strains of S. aureus resistant to CEPs was 6.5% in CEZ, 4.8% in CTM, 8.1% in CXM, and as low as 1.6% in CMZ. PMID- 6285018 TI - [Antibacterial activities of cefotiam against various pathogens isolated from clinical materials (author's transl)]. AB - Bacteriological evaluation was made on cefotiam (CTM), a new broad-spectrum cephalosporin antibiotic, and the following results wer obtained. 1) CTM has shown very potent antibacterial activities against Staphylococcus aureus, Escherichia coli, Edwardsiella tarda, Citrobacter intermedius, Salmonella, Klebsiella, Proteus mirabilis, Proteus rettgeri, Proteus inconstans, Yersinia enterocolitica, Aeromonas hydrophila, Plesiomonas shigelloides and Pseudomonas putrefaciens. 2) Streptococcus faecalis, Enterobacter and Proteus morganii isolated from urine, Serratia and glucose non fermentative Gram-negative bacilli except Pseudomonas putrefaciens, were almost insusceptible to cefotiam. PMID- 6285019 TI - [Antibacterial activities of new cephems against clinically isolated organisms (author's transl)]. AB - The antibacterial activities of cefotaxime (CTX), cefoperazone (CPZ), ceftizoxime (CZX), cefmenoxime (CMX), latamoxef (LMOX), cefotiam (CTM), cefazolin (CEZ), gentamicin (GM) and cefsulodin (CFS) were investigated. All causative organisms were isolated from patients with urinary tract infections treated in Tokai University Hospital. The results were as follows. 1) The MICs of CMX, CTX, and CZX against most of clinically isolated strains of E. coli, K. pneumoniae, Indole (-) Proteus sp. were 0.1 microgram/ml and lower. And then CTM, LMOX and CPZ showed similar antibacterial activities. 2) LMOX and GM showed potent antibacterial activities against C. freundii which was considered to be causative organisms of infections in rare cases. 3) Against S. marcescens, CMX, CTX, CZX, and LMOX showed very potent antibacterial activities. 4) Against P. aeruginosa, CFS, GM and CPZ showed moderate antibacterial activities. 5) Against Enterobacter sp., GM and CMX showed potent antibacterial activities. PMID- 6285020 TI - [Studies on distribution of tissue of cefotiam (author's transl)]. PMID- 6285022 TI - [Concentration of cefotiam in the human bone marrow fluid and bone tissue (author's transl)]. AB - The preoperative administration of cefotiam (CTM) 1 g by intravenous drip infusion for 30 minutes was performed. And the concentration of CTM in peripheral blood, bone marrow fluid and bone tissue was investigated. CTM was smoothly transmigrated from blood to bone tissue, and kept on the concentration in bone tissue over MIC values for several hours. These preliminary results encourage us to think that CTM may be as useful in prophylaxis against postoperative infections. PMID- 6285021 TI - [Distribution of cefotiam dihydrochloride to the intraabdominal exudate (author's transl)]. AB - The concentration of serum and intraperitoneal exudate after dripping intravenous administration of CTM were measured. Four male cases of the emergent laparotomy, a Cushing's ulcer, a hemorrhagic gastric ulcer, an adhesive ileus and acute terminal ileitis were investigated on this study. Two grams of CTM dissolving into 100 ml of physiological saline solution were prepared and administered in a period of 1 hour by intravenous drip (i.v.d.) for all cases after operation. The measurement of CTM-concentration in serum and intraperitoneal exudate were made at 30 minutes, 60 minutes following administration by serum and intraperitoneal exudate were made at 30 minutes, 60 minutes following administration by i.v.d. and at 30 minutes, hourly intervals over a 6-hour period after total administration. The intraperitoneal exudate for this study were collected through a drain which provided at the operation. These samples were measured by Agar-well method at Takeda Central Research Laboratory. Takeda Co., Ltd. The mean value of serum CTM at 30 minutes after the beginning of i.v.d. administration was 65.7 micrograms/ml and the maximum value, 79 micrograms/ml was obtained at 60 minutes. The following mean values at 30 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours and 6 hours after total administration were 39.1, 26.4, 14.6, 8.9, 4.7 and 0.6 microgram/ml. On the other hand, the mean values of CTM intraperitineal exudate at the same time intervals were 4.9, 20.7, 32.0, 34.3, 22.4, 13.9, 11.2, 8.7 and 3.3 micrograms/ml. These results showed that there were much transfer of CTM between serum and intraperitoneal exudate corresponding to the amount of administration. PMID- 6285023 TI - [Biliary tract infections and antibiotics concentration of cefotiam in gallbladder tissue and bile (author's transl)]. AB - In 12 patients undergoing operation, 1 g of cefotiam (CTM) was administered intravenously and CTM levels in gallbladder tissue and gallbladder bile were examined. CTM concentration in gallbladder bile was high in patients with patent cystic duct, while very low in those with cystic duct obstruction. CTM concentration in gallbladder tissue was low in patients with chronic inflammatory gallbladder. CTM activities of T-tube bile were compared with those of CMZ by cross over method in 2 cases. CTM showed extremely higher concentration in bile than CMZ. In a case with excessive output of bile, CTM activity, total bile acids concentration and canalicular flow/ductal flow ratio were low. It can be presumed that CTM was diluted by increased ductal secretion. PMID- 6285024 TI - [Penetration of cefotiam dihydrochloride into cerebrospinal fluid (author's transl)]. AB - It is well recognized that only very low concentration of antibiotics is obtained from cerebrospinal fluid (CSF) despite its high blood concentration. It has been attributed to the blood-brain and blood-CSF barriers. Penetration of CTM into CSF was studied in 7 patients. Two of them were complicated with septic meningitis, and others were not infected. CTM was administered intravenously and samples were obtained from both serum and CSF from 15 minutes to 4 hours for determination of concentration of the antibiotics. In 2 patients with meningitis, the peak level of CTM in CSF after intravenous injection of 2 g and 1 g of CTM was 197 mcg/ml (46% of peak serum concentration), and 17.3 mcg/ml (38% of peak serum level), respectively. In noninfected patients the peak level of CTM in CSF after intravenous injection of 1g of CTM was from 0.3 mcg/ml to 1.9 mcg/ml (0.84% approximately 3.64% of peak serum concentration). We conclude that the percent penetration of CTM into CSF increases in the presence of the inflamed meninges and that prophylactic dosage of CTM for postoperative meningitis will be intravenous administration of 2 g of CTM in adults. PMID- 6285025 TI - [Transportation of cefotiam from serum to cerebrospinal fluid (author's transl)]. AB - Cefotiam (1 g) was administered by one-shot intravenous injection to the patients proceeding a brain surgery. The cerebrospinal fluid (CSF) and serum were taken as a specimen, and the concentration CTM was assayed by agar-well method using Proteus mirabilis ATCC 21100. The most high concentration of CTM was 2,273 micrograms/ml in serum, and 2.3 micrograms/ml in CSF, respectively. The concentration of CTM in serum, CSF, and CSF/serum ration were determined at the indicated time. It appears likely that CTM can pass into CSF more easily than other cephalosporins. PMID- 6285026 TI - [Cefotiam preparations]. PMID- 6285027 TI - [Review [New antibiotics series VI]: Ceftizoxime (author's transl)]. PMID- 6285028 TI - [Concentration of cefotiam in the human bone marrow blood (author's transl)]. PMID- 6285029 TI - [Antibacterial activities of cefotiam against various causative organisms isolated from clinical materials (author's transl)]. AB - Antibacterial activities of cefotiam (CTM), cefazolin (CEZ) and cefmetazole (CMZ) against various causative organisms isolated from clinical materials were investigated. CTM showed excellent antibacterial activities against Gram negative bacilli. Especially against E. coli and Klebsiella, CTM showed superior antibacterial activities to other antibiotics, CMZ and CEZ. The MICs of CTM against Serratia and Enterobacter dispersed very widely, but showed small values comparing with CMZ and CEZ. The antibacterial activity of CTM against P. mirabilis was slightly superior to that of CMZ, but against P. vulgaris was rather inferior to that of CMZ. However, these 3 antibiotics, CTM, CMZ and CEZ, showed slightly less active against non-glucose-fermentation bacilli. PMID- 6285030 TI - [Fate of Clostridium perfringens in intestine of guinea pigs after oral ingestion (author's transl)]. PMID- 6285031 TI - [A case of congenital pure red cell anemia, accompanied by thenar defect, syndactyly, and cardiovascular anomalies (author's transl)]. PMID- 6285032 TI - [Laparoscopy and biopsy]. PMID- 6285033 TI - [Metabolic system of blood platelets and thrombus formation]. PMID- 6285034 TI - [Chemistry of sodium potassium ATPase]. PMID- 6285036 TI - [Cytochemical bioassay (CBA) of hormones]. PMID- 6285035 TI - [Immunoglobulin genes and B cell differentiation, with special reference to neoplastic changes of B cells]. PMID- 6285037 TI - [The CDC survey of high density lipoprotein cholesterol measurement and our results (author's transl)]. PMID- 6285038 TI - [Ultrasonic diagnosis of tumor thrombosis of the portal vein and inferior vena cava (author's transl)]. PMID- 6285039 TI - [The clinical significance of computed tomographic arteriography for minute hepatocellular carcinoma (author's transl)]. PMID- 6285040 TI - [A case of liver cell adenoma (author's transl)]. PMID- 6285042 TI - [Various liver diseases and antibody responses to cytomegalovirus (author's transl)]. PMID- 6285041 TI - [An autopsy case of hepatic cell carcinoma surviving for two and a half years after "embolization therapy" (author's transl)]. PMID- 6285043 TI - [Clinicopathological evaluation of angiograms in pancreatic and hepatocellular carcinoma (author's transl)]. PMID- 6285044 TI - [Study on epoxide hydrolase in rat hyperplastic nodule and hepatoma during chemical hepatocarcinogenesis induced by 2-acetylaminofluorene (author's transl)]. PMID- 6285045 TI - [Effect of polymyxin B on endotoxemia in severe liver diseases (author's transl)]. PMID- 6285046 TI - [Ultrastructural differences in three types of the lymphocytic cytotoxicities against Chang liver cells and the human hepatoma cell line in vitro (author's transl)]. PMID- 6285047 TI - The effect of Bordetella pertussis lymphocytosis-promoting factor (LPF) on antibody response in mice: its enhancing and suppressive effects. AB - The effect of lymphocytosis-promoting factor (LPF) on antibody response in mice was estimated under different sets of experimental conditions. Four- and 6-week old mice were intravenously inoculated with LPF. Three days later these mice were inoculated either intraperitoneally or intravenously with sheep red blood cell (SRBC) or human serum albumin (HSA) as an antigen. The adjuvant effect of LPF was demonstrated on antibody response in 6-week old mice to intraperitoneally inoculated SRBC but not to intravenously-inoculated one. When 4-week-old mice were immunized, hemagglutinin production in response to intraperitoneally inoculated SRBC was not enhanced by LPF. In addition, a rather suppressive effect of LPF at a comparatively high dose was demonstrated on hemagglutinin production in response to intravenously inoculated SRBC. Anti-HSA production was enhanced by inoculation of LPF in any combination of the mouse age and the route of antigen administration. These findings indicate that the adjuvant effect of LPF on antibody response in mice depends upon experimental conditions: the age of mice, the quality of antigen and the route of antigen administration used for immunization. PMID- 6285048 TI - Effects of local anesthetics on monoamine oxidase, and their membrane effects. AB - The effects of various local anesthetics on rat brain and liver monoamine oxidase (MAO) and their antihemolytic and local anesthetic effects were studied. All local anesthetics tested at 1 x 10(-7) M to 1 x 10(-3) M inhibited MAO activity in rat liver mitochondria with 5-hydroxytryptamine (5-HT) as substrate. The order of potency was tetracaine>procaine>dibucaine>lidocaine>prilocaine. Tetracaine and procaine inhibited 5-HT oxidation much more than beta-phenylethylamine (PEA) oxidation. Dibucaine inhibited PEA oxidation as much as 5-HT oxidation. Inhibition of MAO by local anesthetics other than dibucaine was reversible. Tetracaine and procaine inhibited 5-HT oxidation competitively, whereas dibucaine inhibited it non-competitively. Antihemolytic effects were observed with dibucaine and tetracaine at concentrations of 6 x 10(-5) M and 1 x 10(-4), respectively. The order of surface anesthetic potencies was dibucaine>tetracaine>prilocaine>lidocaine>procaine. These results suggest that the inhibition of MAO activities by local anesthetics depends on both electrostatic and hydrophobic interactions between these drugs and enzyme associated phospholipids or the hydrophobic regions of proteins. PMID- 6285049 TI - Influence of hypothyroid status on dopamine-induced positive chronotropic and inotropic effects on isolated rat atria. AB - The involvement of alpha- and beta-adrenoceptors in dopamine (DA)-induced positive chronotropic and inotropic effects was investigated in isolated atria from euthyroid and hypothyroid rats. Propranolol at 3 x 10(-7) M remarkably inhibited the positive chronotropic effect of DA in both euthyroid and hypothyroid rats, and 1 x 10(-6) M phentolamine inhibited the effects of DA in the presence of propranolol in hypothyroid rats. Propranolol remarkably inhibited the positive inotropic effect of DA in both euthyroid and hypothyroid rats, while phentolamine was effective only in hypothyroid rats. In atria from reserpinized rats, the pD2-values for DA in both chronotropic and inotropic effects were reduced, but effectiveness of propranolol or phentolamine on DA-induced positive chronotropic and inotropic effects in euthyroid and hypothyroid rats was similar to that in non-reserpinized rats. In hypothyroid rats, DA increased the maximal rate of tension development, which was inhibited by both phentolamine and propranolol. DA shortened the duration of contraction. DA in the presence of phentolamine significantly shortened the duration of contraction but did not in the presence of propranolol. In conclusion the DA-induced positive chronotropic effect is mainly produced by beta-adrenoceptor stimulation in both euthyroid and hypothyroid rats, and also by a alpha-adrenoceptors to some extent in hypothyroid rats. The DA-induced positive inotropic effect is produced by alpha- as well as beta-adrenoceptor stimulation in both groups. However, alpha-adrenoceptors were involved to a greater extent in hypothyroid rats than in euthyroid rats. The stimulation of alpha-adrenoceptors by DA causes an increase in the maximal rate of tension development without a significant change in the duration of contraction, and the stimulation of beta-adrenoceptors by DA causes an increase in the maximal rate of tension development and shortening of the duration of contraction. PMID- 6285050 TI - Anti-asthmatic activity of BB-1502 by inhalation. AB - The anti-asthmatic activity, pulmonary mechanics and cardiovascular effects of BB 1502 (9-cyclohexyl-2-n-propoxy-9H-adenine) administered by aerosol were evaluated in guinea pigs and dogs and compared with the effects of salbutamol. BB-1502 protected guinea pigs from allergic asthma induced by aerosolized egg albumin antigen. The potency of BB-1502 was approx. 1/7 that of salbutamol when the test drugs and antigen were aerosolized simultaneously, whereas the two drugs were equipotent when administered 60 min prior to the antigen challenge. The duration of the anti-asthmatic action of BB-1502 appeared to be much longer than that of salbutamol. In dogs, BB-1502 and salbutamol showed comparable activity in preventing ascaris antigen-induced asthmatic symptoms. Disodium cromoglycate showed anti-asthmatic activity at high doses while aminophylline was inactive by inhalation. No cardiopulmonary side effects were observed in guinea pigs or dogs following the inhalation of BB-1502. The doses were 800-5,000 times higher than those needed to produce the anti-asthmatic activity. Salbutamol caused significant tachycardia, hypotension and tachypnea at doses 100-300 times its anti-asthmatic dose. PMID- 6285051 TI - Effects of anti-asthmatic drugs on airway resistance and plasma level of cyclic AMP in guinea pig. AB - Effects of anti-asthmatic drugs on airway resistance and plasma level of cyclic AMP were investigated in guinea pigs sensitized and non-sensitized with egg albumin. Histamine increased airway resistance in the both groups of guinea pigs, and guinea pigs sensitized with egg-albumin were more sensitive to histamine. Anti-asthmatic drugs inhibited dose-dependently the increase of airway resistance caused by histamine. Sensitization with egg-albumin decreased the potencies of the beta-adrenoceptor stimulants, salbutamol and isoprenaline, but not that of aminophylline. The plasma level of cyclic AMP was increased by salbutamol and isoprenaline, but not by aminophylline. The increased plasma level of cyclic AMP by beta-adrenoceptor stimulants was not attenuated by sensitization with egg albumin. PMID- 6285052 TI - [A case of allergic bronchopulmonary aspergillosis induced by bone powder, a fertilizer (author's transl)]. PMID- 6285053 TI - [Heterotransplantation of human malignant urogenital tumors. (3) Basic studies of transplantable human prostatic carcinoma in nude mice and susceptibility testing of several platinum compounds (author's transl)]. PMID- 6285054 TI - Spontaneous nephroblastoma in 3 Sprague-Dawley rats. PMID- 6285055 TI - Studies on Getah virus: some biological, physicochemical and antigenic properties of the MI-110 strain. PMID- 6285056 TI - Enteritis due to feline infectious peritonitis virus. PMID- 6285057 TI - T-lymphocyte and B-lymphocyte subpopulations infiltrating human mammary carcinomas. AB - Both T- and B-lymphocytes were found in human primary mammary carcinomas and were distributed in widely varying amounts, but in most tumors, T-lymphocytes predominated. A small percentage of the T-lymphocytes expressed receptors for the Fc portion of IgG (Fc gamma R), but very few had receptors for C3 (C3+) (comparable to the findings in blood). A prominent subset of lymphocytes had Fc gamma R and were C3+, and most of these were surface immunoglobulin (slg)-bearing cells. The majority of lymphocytes from this subset were Fc+ C3+, and only a small percentage were Fc+ C3- (in contrast to the findings in blood). IgD and IgM were the predominant classes of findings in blood). IgD and IgM were the predominant classes of immunoglobulin found on the B-lymphocytes. The different preparative techniques did not result in a selective loss of lymphocyte subsets, but collagenase digestion did lead to a loss of expression of the C3 receptor on the lymphocyte surface, which was recoverable when lymphocytes were reincubated at 37 degrees C. No evidence was found for blocking of the C3 receptor by immune complexes with activated complement. PMID- 6285059 TI - Tumor virus induction and host cell capacity inactivation: possible in vitro tests for photosensitizing chemicals. AB - The responses of two in vitro mammalian virus-host cell systems to the photosensitizing chemicals proflavine sulfate and 8-methoxypsoralen (8-MOP) in the presence of light are described. Infectious simian virus 40 (SV40) could be induced from SV40-transformed hamster cells by treatment with proflavine plus visible light or 8-MOP plus near UV radiation. The same photosensitizing treatments inactivated the capacity of monkey cells to support the growth of herpes simplex virus. SV40 induction and inactivation of host cell capacity for herpesvirus growth might be useful as screening systems for testing the photosensitizing potential of chemicals. Advantages and disadvantages associated with each system are discussed. PMID- 6285058 TI - Molecular aspects of drug photosensitivity with special emphasis on psoralen photosensitization reaction. AB - Photosensitization reactions involve phototoxic reactions and photoallergic reactions, which are less common. Common drugs and chemicals that photosensitize humans are listed. Phototoxic reactions include the following: 1) direct photosensitization (type I reactions), in which the reactions of the triplet state sensitizer are directly attributable to a component other than oxygen (e.g., DNA, proteins, and cell membranes), and 2) indirect photosensitization (type II reactions), in which the triplet state of a sensitizer reacts first with molecular oxygen, producing an "active oxygen" intermediate that subsequently reacts with the biologic system. The active oxygen intermediates are singlet oxygen (1O2)k, superoxide radical anions (O(2), and hydroxy radicals. Psoralen induced skin photosensitization appears to involve both type I and type II reactions. The formation of monofunctional bifunctional psoralen-DNA photoadducts (a type I reaction) is probably responsible for cell damage, cell death, mutation, and even skin carcinogenesis. The erythema response appears to be the result of a type II reaction. PMID- 6285060 TI - Interaction of oxygen and oxy-radicals with carotenoids. AB - Dye-sensitized photo-oxidations of liposomes are inhibited by the presence of carotenoids in the membrane. Some carotenoid bleaching occurs during the reaction. It was shown that carotenoids are capable of inhibiting free-radical induced lipid peroxidation in liposomes as well as singlet oxygen-induced lipid peroxidation. There is little direct effect of lipid peroxides on carotenoid bleaching during the photo-oxidative reactions. The carotenoid bleaching observed during the dye-photosensitized oxidations of the liposomes appears to be due to interactions between the carotenoid pigments and either radical intermediates from type I reactions or direct chemical reaction with singlet oxygen in a type II reaction. It was concluded that carotenoid pigments function as protective agents by quenching triplet sensitizers, singlet oxygen, and radical intermediates. PMID- 6285062 TI - Ocular phototoxicity and psoralen plus ultraviolet radiation (320-400 nm) therapy: an experimental and clinical evaluation. AB - The increase in fluorescent chromophores within the human ocular lens is age related, leading to an increasingly yellow core (nucleus) that presumably results from UV radiation exposure. In approximately 10% of our population this process progresses more rapidly, resulting in the formation of the brown (nuclear) cataract. Some lenticular discoloration may be beneficial, since it enables the mature lens to filter UV and short-wavelength visible radiation, thus protecting the retina from potential photodamage. Aphakic primate retinas can be irreversibly damaged by exposure to approximately 5 mW.cm-2 long-wavelength UV (greater than 325 nm) radiation. Photosensitized damage to the lens and retina with psoralen plus UV radiation (320-400 nm) (PUVA) has been demonstrated in experimental animals, and cataracts have recently been reported in patients given PUVA therapy. A new method to screen patients for lens damage is by enhanced fluorescence measurements. This method, UV slit-lamp densitography, permits detection of lenticular photodamage at a molecular level, years before visible opacities become manifest by conventional slit-lamp examination. This procedure has also demonstrated a significantly lower level of lens fluorescence (hence decreased filtering capacity) in patients with retinal degenerative diseases, suggesting UV photodamage as a factor in the progression and perhaps pathogenesis of these conditions. PMID- 6285061 TI - Circulating immune complex levels in patients with gestational trophoblastic neoplasia. AB - The clinical course, human chorionic gonadotropin (HCG) levels, and serial circulating immune complex (CIC) levels in 21 patients with gestational trophoblastic neoplasia (GTN) were correlated for the evaluation of the relationship between CIC levels and trophoblastic tumor burden. CIC levels were normal in 18 of 21 patients at the time of presentation, and 2 of 3 patients who presented with elevated CIC levels had significant comorbid disease (toxemia and hepatitis). Nine patients were followed into gonadotropin remission, and all 9 developed an increase in CIC levels at the time of remission. It was concluded that CIC, at least as measured by two antigen-nonspecific techniques, is generally not elevated at initial presentation in the patient with GTN; this lack of an elevation is probably due to marked tumor antigen excess. Thus the in vivo importance of CIC as a "blocker" of host antitumor response at this stage is doubtful. After effective treatment as HCG levels return to normal, the demonstrated elevation in serial levels of CIC may reflect a return of adequate host immune response at a time of minimal tumor burden. PMID- 6285063 TI - Steroid hormone receptors in the rat mammary adenocarcinoma induced by N-hydroxy N-2-fluorenylacetamide. AB - The steroid hormone receptors in N-2-fluorenylacetamide (2-FAA)- and N-hydroxy-2 FAA-induced mammary adenocarcinomas in outbred Sprague-Dawley rats were analyzed. Both 8S and 4S estrogen-binding components have been detected in cytosols of these tumors following sucrose gradient sedimentation in low salt. Competitive binding analyses of this binder indicated a specificity profile expected of an estrogen receptor. Both androgen and progesterone receptors were also present in the cytosols of these mammary tumors. While the androgen receptor sedimented in the 8S region of the gradient, the progestin binder appeared only as a 4S moiety under similar conditions. The relative concentrations of these receptors (expressed in fmol/mg protein +/- SE) were: 17 beta-estradiol (28.6 +/- 4.1) greater than 5 alpha-dihydrotestosterone (8.5 +/- 2.2) greater than progesterone (5.0 +/- 1.3). The progesterone receptor was increased at least eightfold in the mammary adenocarcinomas from ovariectomized rats that were treated with diethylstilbestrol for 6 days. Binding equilibrium data indicated Ka = 1.2-1.8 X 10(9) M-1 for the above cytoplasmic hormone receptor complexes (Ka, association constant). Although cytosols prepared from lactating mammary gland contained appreciable quantities of glucocorticoid receptor, only trace amounts were found in the mammary adenocarcinoma. PMID- 6285065 TI - [Role of stress in the pathogenesis of ischemic heart disease]. PMID- 6285064 TI - [Drugs blocking the calcium channel]. PMID- 6285066 TI - Renal concentration defect following nonoliguric acute renal failure in the rat. AB - The mechanism of impaired renal concentrating ability following nonoliguric ischemic acute renal failure was studied in the rat. Fifty min of complete occlusion of the renal artery and vein with contralateral nephrectomy resulted in reversible, nonoliguric acute renal failure. Eight days following induction of acute renal failure, a defect in 30 hr dehydration urine osmolality was present when experimental animals were compared with uninephrectomized controls (1,425 +/ 166 versus 2,267 +/- 127 mOsm/kg water respectively, P less than 0.001). Comparable postdehydration plasma vasopressin levels in experimental and control animals and an impaired hydro-osmotic response to exogenous vasopressin in experimental animals documented a nephrogenic origin of the defect in urine concentration. Lower urinary excretion of prostaglandin E2 in experimental animals and a failure of cyclo-oxygenase inhibition with 10 mg/kg of indomethacin to improve dehydration urine osmolality suggested that prostaglandin E2 antagonism of vasopressin action did not contribute to the concentration defect. Postdehydration inner medullary (papillary) interstitial tonicity was significantly reduced in experimental animals versus controls (870 +/- 85 versus 1,499 +/- 87 mOsm/kg water respectively, P less than 0.001). To determine if this decreased interstitial tonicity was due to vascular mechanisms, papillary plasma flow was measured and found to be equivalent in experimental and control animals. To examine a role for biochemical factors in the renal concentration defect, cyclic nucleotide levels were measured in cytosol and membrane fragments. A decrease in vasopressin and sodium fluoride-stimulated adenylate cyclase was found in outer medullary tissue of experimental animals. In contrast, vasopressin stimulated adenylate cyclase activity was comparable in the inner medullary tissue of control and experimental animals. Our study suggests a defect in generation of renal inner medullary interstitial solute as a mechanism of the impaired urinary concentration observed in this model of acute renal failure. PMID- 6285067 TI - [Effect of cryotherapy application to burned cornea on collagenase activity in organtypic culture (author's transl)]. PMID- 6285068 TI - [Cryotherapy effect on collagenase activity inhibition in experimental corneal penetrating wounds (author's transl)]. PMID- 6285069 TI - [Effect of more modern drugs on corneal epithelium regeneration (author's transl)]. PMID- 6285072 TI - An electronmicroscopic study on the malignant lymphomas of SL/K, SL/Ni and hybrid of SL/K and SL/Ni mice. PMID- 6285070 TI - [Physiological and pathophysiological significance of superoxide-radicals and the regulatory role of the enzyme superoxide dismutase (author's transl)]. AB - The monovalent reduction of molecular oxygen, resulting in the formation of superoxide radicals (O(2)) is regarded as to be an ongoing physiological process involved in the respiration and other biological processes of aerobic cells. These reactive oxygen species have been reported to function as cofactors in many biosynthetic reaction steps. Thus, deviations from cellular steady state concentrations may lead to a multiplicity of clinical symptoms or may to a great deal determine the characteristic of a distinct malady. Decrease of cellular O(2) concentration is discussed in connection with Trisomie 21 and various mental disorders. The role of O(2) in the biochemistry of inflammation, autoimmune diseases, various toxicological cases and the biological aging process is described. Hypothetical considerations concerning the involvement of O(2) in the pathogenetic mechanisms of Morbus Wilson, haemochromatosis, Parkinson syndrome, cataractogenesis and in carcinogenesis are presented. The physiological control of cellular O(2)-concentration is performed by formation rates of the various cellular O(2)-sources and the overall elimination rates of O(2)-consuming reaction steps. Superoxide dismutase (SOD) is of special interest within this cycle because it detoxifies O(2) radicals with velocity rates which are significantly faster than any other pathway involved in O(2) elimination. Thus attempts for a therapeutic interference on tissue levels of O(2)-radicals are mainly based on inhibition or activation of cellular SOD-activities depending on a supposed decrease or increase in cellular steady state concentrations of O(2). The availability of a drug version of SOD and of various synthetic SOD-active compounds allowing a therapeutic decrease of O(2)-tissue levels. Inhibition of cellular SOD is also possible, however, many still unknown toxic side effects should be expected because of unspecific action of the inhibitor available. PMID- 6285071 TI - Ferrokinetics after high-dose methotrexate therapy. AB - After high-dose methotrexate with doses ranging from 2-10 g/m2, ferrokinetics were performed in 11 patients with different tumors. Iron-III-citrate-59Fe was injected at methotrexate serum concentrations ranging between 2.7 x 10(-7) -1.3 x 10(-8) M. With MTX levels greater than or equal to 4.2 x 10(-8) M the plasma iron clearance was always retarded, and the plasma iron turnover was diminished in 5 of 6 patients with levels of 3.1-8.9 x 10(-8) M at the time of the injection. In all cases with MTX concentrations greater than or equal to 5 x 10(-8) M the 59Fe utilization as the measure of effective erythropoiesis was reduced pathologically below normal range. These results show that the erythropoiesis resumes its normal extent even in case of normal methotrexate clearance only when the methotrexate serum concentrations have fallen down below 5 x 10(-8) and that erythropoiesis is more sensitive against methotrexate toxicity than the granulocytopoiesis. PMID- 6285073 TI - Studies on glycogen phosphorylase kinase: catalytic properties and isozymes. PMID- 6285074 TI - Total and unbound bilirubin determination using an automated peroxidase micromethod. PMID- 6285075 TI - [Indices of natural body resistance in viral hepatitis]. PMID- 6285076 TI - [Role of hormonal compounds in regulating electrolyte metabolism in emotional stress]. AB - The emotional stress simulated by examinations taken by medical students led to a decrease of sodium concentration in blood. This was a result of a simultaneous increase of mineralocorticoids and a decrease of glucocorticoids responsible for sodium concentration in blood. Both inhibition of glucocorticoids and stimulation of mineralocorticoids were associated with a high activity of plasma renin and a low concentration of adrenocorticotropic hormone. PMID- 6285078 TI - Effects of experimental infection of the deer mouse (Peromyscus maniculatus) with mouse hepatitis virus. AB - Three deer mice (Peromyscus maniculatus) given 10(4) TCID50 of mouse hepatitis virus -S intranasally all had significant serum neutralization titers to mouse hepatitis virus -S 3 weeks later. Ten other deer mice were given 10(3) TCID50 of mouse hepatitis virus -3 intranasally and placed with sentinel laboratory mice (CF1, C3H/Rv, and nu/nu) and uninoculated deer mice. One inoculated deer mouse died, but mouse hepatitis virus was not isolated. No other inoculated or uninoculated animals exhibited signs of disease. No sentinel animals exhibited serum neutralization titers for mouse hepatitis virus -3. Three of the remaining nine inoculated deer mice had positive antibody titers. It was concluded that deer mice do not develop clinical manifestations of mouse hepatitis virus or transmit the virus to potentially susceptible laboratory mice. PMID- 6285077 TI - Implication of Clostridium difficile and Clostridium perfringens iota toxins in experimental lincomycin-associated colitis of rabbits. AB - Following oral administration of lincomycin, over 50% of two groups of 12 rabbits each died between 4 and 56 days with distended, non-hemorrhagic, fluid-filled ceca. Bacteria-free cecal filtrates from the rabbits that died were lethal for mice, cytopathic in Y-1 tissue culture monolayers, and caused increased vascular permeability in rabbit skin. Although the cecal filtrates of both groups had similar biological activity, the filtrate activity of one group was neutralized by Clostridium perfringens iota antitoxin, and the infiltrate activity of the other group was neutralized by Clostridium difficile antitoxin. Toxigenic Clostridium difficile also were isolated from the ceca of this group. A broth filtrate of a Clostridium difficile culture was lethal for rabbits when injected intravenously and intraperitoneally. The data indicated Clostridium difficile toxin or clostridium perfringens iota toxin may be associated with lincomycin associated colitis in rabbits. PMID- 6285080 TI - Rotavirus (SA11) antibody in nonhuman primates. PMID- 6285079 TI - Nephroblastoma with secondary polycythemia in a New Zealand white rabbit. PMID- 6285081 TI - Nerve abscess in lepromatous leprosy. PMID- 6285082 TI - Prostacyclin increases gastric mucosa blood flow via cyclic AMP. PMID- 6285083 TI - Dissociation of aldosterone response to angiotensin II and potassium in the presence of strontium. AB - the effect of Ca2+, Sr2+ and Ba2+ on aldosterone output was studied in basal conditions and under the stimulus of angiotensin II, potassium and ACTH. Aldosterone production was measured in glomerulosa cells isolated from bovine adrenal glands and incubated in Krebs-Ringer solution in which 1.25 mM Ca2+ was substituted by Sr2+ or Ba2+. Strontium and Ba2+ can substitute for Ca2+ in steroid production when angiotensin II or ACTH were the aldosterone-stimulating agents. The relative aldosterone production was Ca greater than Sr greater than Ba. However, when the steroidogenic effect of potassium was measured under similar conditions, neither Sr2+ nor Ba2+ were able to replace Ca2+ responses. The results suggest a dissociation between the calcium mediated stimulation of aldosterone production by angiotensin II and potassium. PMID- 6285084 TI - [Comparative effects of an extract of Ginkgo biloba and chlorpromazine on the in vitro osmotic fragility of rat erythrocytes (author's transl)]. AB - The effects of different concentrations of an extract of Ginkgo biloba (Gb) and of chlorpromazine (CPZ) on the osmotic lysis of rat erythrocytes (produced by dilution of the incubation medium by 30%-70%) were compared in vitro. The erythrocytes were incubated together with Gb or Cpz at the time of the hemolytic reaction at 25 degrees or 37 degrees C in either a simple phosphate buffer or in a more complete buffer containing inorganic salts at physiological concentrations. CPZ produced a dose-dependent increase in the membrane resistance, the maximum effect occurring at 10(-4) M regardless of the experimental conditions used. Gb also increased membrane resistance, but to a lesser extent than CPZ. In addition, Gb was more effective under conditions which increased erythrocyte membrane fragility; i.e., 37 degrees C, simple phosphate buffer. The mechanism of action of Gb seems to differ from that of CPZ. The activity of CPZ (as a membrane stabilizer) is, in part, due to its penetration into the membrane phospholipid layer. Gb might act in a similar manner, but could also modify Na+ transport across the cell membrane by an action on adrenergic receptors, or could stimulate Na+, K+-ATP-ase activity, as has been described for norepinephrine. As Gb contains many molecular constituents: its action on the membrane is likely to be quite complex; but this effect could be associate with its vascular therapeutic action. PMID- 6285085 TI - Pharmacological inhibition of calmodulin-sensitive phosphodiesterases. AB - Two cyclic nucleotide phosphodiesterases sensitive to calmodulin were partially purified from bovine brain and bovine aorta. The enzyme from bovine brain exhibited the properties of the "high Km phosphodiesterase", with a higher Km for cyclic AMP than for cyclic GMP. The enzyme from bovine aorta hydrolyzed both cyclic AMP and cyclic GMP with similar kinetics. The hydrolysis of cyclic AMP by the brain enzyme was selectively inhibited by papaverine and 2' deoxy cyclic AMP. The hydrolysis of cyclic GMP by this enzyme was selectively inhibited by cyclic IMP, 2' deoxy cyclic GMP, 1-methyl-3-isobutyl-xanthine, ICI 74917 and M & B 22948. All these inhibitors were identically potent on the hydrolysis of both substrates by the aorta enzyme. Ro-20 1724, an imidazolidinone, and ZK 62711, a pyrrolidone, two compounds known to be potent inhibitors of cyclic AMP phosphodiesterase insensitive to calmodulin, were inefficient inhibitors of both calmodulin-sensitive phosphodiesterases. PMID- 6285086 TI - Clonidine inhibits diuresis caused by negative pressure breathing and left atrial distension: role of central adrenergic alpha 2 receptors. AB - 1. Intravenous (10 micrograms/kg) or intracisternal (1 microgram/kg) clonidine inhibited the diuretic response elicited by negative pressure breathing (NPB) or left atrial distension (LAD) in chloralose anaesthetized dogs. The drug reduced the induced tachycardia, but not the increase in respiratory rate elicited by NPB. Blood pressure was unchanged. 2. Propranolol (1 mg/kg IV) did not change NPB induced diuresis. 3. Intravenous yohimbine (1 mg/kg IV) restored these two diuresis inhibited by intravenous or intracisternal clonidine, whereas prazosin (0.05 mg/kg IV) was without effect. 4. These results show that the adrenergic receptor implicated in the volumetric control of vasopressin secretion could be of the alpha 2 subtype. This alpha 2 adrenoceptor could be localized at a central level. 5. The possible clinical consequences (hydrosaline retention and antihypertensive drugs; use of clonidine in aerospace medicine) of these data are discussed. PMID- 6285087 TI - Quantification, smoothing, and confidence limits for single-units' histograms. AB - In this article the relationships among firing rate, probability of firing and counts per bin are examined. It is suggested that PSTHs, autocorrelations and crosscorrelations of neuronal activity should all be expressed in units of firing rates (spikes/s), since the values obtained by such scaling are independent of bin size and of total time of measurement. A simple method for these histograms is described. Methods to compute confidence limits for PSTHs, autocorrelations and crosscorrelations are suggested. The computations are based on the null hypothesis that the spike train(s) is (are) the realization of (independent) Poisson-point process(es). The validity and the limitations of these computations methods, when applied to spike trains, are discussed. Methods to smooth out random fluctuation with little distortion of the histogram's shape are described. It is suggested that one can minimize the distortion of the histogram in the time domain and in the frequency-domain by using a bell-shaped bin whose center point slides continuously along the histogram. The article aims at giving the potential user of the methods some insight for the meaning of the formulae. It describes in detail how the methods are applied in practice and illustrates each method by using real data from single-unit recordings. PMID- 6285088 TI - A micromethod for the determination of ceramide. AB - An enzymatic micromethod for the determination of ceramide is presented. The enzyme, E. coli diglyceride kinase was used to phosphorylate ceramide, as well as diglyceride with high specific activity gamma-[32P]ATP, and the two products are differentiated by their alkali stability. This method was applied to the detection of endogenous phospholipases and sphingolipases in several membrane systems and could have wide application. PMID- 6285090 TI - EB-virus and rheumatoid arthritis: new insights into their interrelation. PMID- 6285089 TI - The management of lung cancer. PMID- 6285091 TI - [Dietary fiber]. PMID- 6285092 TI - Therapy of superficial fungal infection. PMID- 6285093 TI - [Esophageal rupture after intake of natural products]. PMID- 6285094 TI - [Total stop in esophagus after intake of natural products]. PMID- 6285095 TI - [Therapy of beta-cell nesidioblastosis in childhood (author's transl)]. PMID- 6285096 TI - [On the viral etiology of Bell's palsy (An enzyme-linked immunosorbent assay study) (author's transl)]. AB - In a prospective study paired sera of 14 patients suffering from Bell's palsy were examined for antibodies against Varicella-zoster Virus (VZV), Herpes-simplex Virus (HSV), Cytomegalovirus (CMV) and Epstein-Barr Virus (EBV). For determination of antibodies against VZV, HSV and CMV an enzyme-linked immunosorbent assay was carried out. Indirect immunofluorescence was carried out to determine antibodies against EBV. Serum samples were taken within 5 days of Bell's palsy having been diagnosed and were compared with further serum samples taken 4 weeks later. No evidence of significant differences between the antibody titers of the paired sera was found. The viral etiology of Bell's palsy due to an exogenous infection or by activation of a latent infection seems unlikely. PMID- 6285097 TI - [Diagnostic problems concerning glomus jugulare-tumours (author's transl)]. PMID- 6285099 TI - [Postoperative care of patients with chronic pancreatitis (author's transl)]. AB - 137 patients, who had had surgery of the pancreas, were followed up postoperatively; 72 of these had had resection of pancreas, 65 patients had organ preserving surgery. Diabetes occurred in 44% of the total group. Control of diabetes was adequate only in 19%. 89 patients had an insufficient exocrine pancreatic function; substitution with pancreatic enzyme products was not adequate in 39 patients in this group. 65% took up work again, the working ability being reduced by less than 33%. 14% of the whole group retired immediately after operation from working life, 6% remained jobless for longer periods of time. After surgery of the pancreas patients should be encouraged to active work; granting pensions too generously will increase the relapse rate of alcoholism. In 6 cases of subtotal surgery and in 21 cases of organ preserving surgery operation had to be done for a second time. A classification is proposed for evaluation of postoperative results. PMID- 6285098 TI - [Problems after surgery of the colon (author's transl)]. AB - After surgery of the colon 3 types of sequelae may be observed: 1. Acute intra- and postoperative complications, as insufficiency of the anastomosis, ileus, and rebleeding; sequelae of these acute complications may be stenosis of the stenosis and intraabdominal adhesions. 2. Chronic postoperative syndromes like blind-loop syndrome, diarrhoea and/or continence problems, problems arising from an anus praeter, disturbances of sexual and bladder function. These symptoms occur rather rarely after partial resection of the rectum or amputation of the rectum. 3. Recurrence of the main disease, especially cancer disease, be it a local recurrence or metastasis. A well organized follow-up of cancer patients may increase survival time only in a few cases, but it definitely may improve quality of life by different means. PMID- 6285100 TI - [Piracetam and the activity of alpha- and beta-adrenergic, dopaminergic and muscarine cholinergic receptors (author's transl)]. PMID- 6285101 TI - Identification of gamma-aminobutyric acid and its binding sites in the rat ovary. AB - Gamma-aminobutyric acid (GABA), GABA synthesizing enzyme and GABA binding sites were measured in rat ovaries. The concentration of GABA in the ovary (0.56 microgram/mg protein) was less than that in the brain (1.2-3.4 microgram/mg protein), but was six-fold higher than any other non-neuronal tissue examined. Glutamate decarboxylase, the GABA synthesizing enzyme was also found in high concentrations in whole ovarian homogenate but not in enriched ovarian granulosa cells, testis, anterior pituitary or muscles. Furthermore, high affinity (Kd = 15 21 nM), specific GABA binding sites were identified in the ovaries by specific [3H]muscimol binding and the majority of GABA binding sites were associated with the granulosa cells. These data suggest a possible role of GABA in the regulation of ovarian functions. PMID- 6285102 TI - Neurochemical and behavioral correlates of antidepressant drug action. AB - Chronic (4 days or more) administration of imipramine or mianserin, but not atropine, leads to an extinction in muricidal behavior in the rat. Moreover, receptor binding assays revealed that there is a significant decline in the number of beta-adrenergic, but not serotonin2, receptors in the frontal cortex at the onset of the behavioral modification. While the antidepressants also induced receptor binding changes in nonmuricidal control animals, the pattern of these changes differed from that observed in the muricidal subjects suggesting that the receptor modification was, to some extent, trait-dependent. These findings indicate that, with the muricidal model, chronic rather than acute drug treatment may be a more selective test for antidepressant efficacy. In addition, the data suggest that a decline in brain beta-adrenergic receptors may be causily related to the behavioral modification. PMID- 6285104 TI - Effects of naloxone on basal and stress-induced prolactin secretion, in intact, hypothalamic deafferentated, adrenalectomized, and dexamethasone-pretreated male rats. AB - The effects of naloxone (Na1) on basal and stress-induced PRL secretion were investigated in intact (N) adult male rats, as were its effects in rats with complete hypothalamic deafferentation (CHD), in adrenalectomized (adrenX) rats, and in rats pretreated with dexamethasone (dex). Forty-five minutes subsequent to Na1 administration (5 mg/kg, BW, IP) basal serum levels of PRL were reduced by approximately 25% (p less than 0.05), in both N and CHD groups. PRL secretory responses to acute exposure to both photic and acoustic stress wee markedly attenuated in Na1-injected, as compared to vehicle-injected animals. Basal serum PRL concentrations were elevated by 40% in adrenX rats (p less than 0.05), as reduced by 25% (p less than 0.05) in dex-treated rats, as compared to controls. In both these experimental groups, Na1 administration caused significant reductions in serum PRL. This study demonstrates that stress-induced, as well as basal PRL secretion, is attenuated by Na1, and points to a hypothalamic site of action in this regard. Furthermore, these Na1 effects are independent of glucocorticoid interactions with the CNS. PMID- 6285103 TI - Benzodiazepine antagonists abolish electrophysiological effects of sodium valproate in the rat. AB - A hypothesis was considered that anti-epileptic potency of sodium valproate (VPA) may be associated with its action via the benzodiazepine system. The ability of anti-petit mal drugs to suppress the slow secondary negative wave (SNW) of the visually evoked potential was used as a sensitive electrophysiological "tag" for comparison of VPA (200 mg/kg, i.p.) and Diazepam (5 mg/kg, i.p.) effects. Both drugs induced a profound inhibition of the SNW. Benzodiazepine antagonists Ro 5 3663 (2 mg/kg, i.p.) and Ro 15-1788 (5 mg/kg, i.p.) caused recovery of the SNW amplitude within several minutes of injection. Both antagonists abolished immobility and sedation produced by VPA and Diazepam. The possibility should be considered that therapeutic effects of VPA are mediated through the benzodiazepine receptor coupled to GABA. PMID- 6285105 TI - The prevention of an artifact in receptor binding assay by an improved technique. AB - The intensity of vacuum suction used during filtration of cerebral membranes on glass fiber filters used in the assays of neurotransmitter receptors is a factor in determining the apparent extent of bound radioactivity. This is not related to protein loss during filtration at various speeds nor to rates of dissociation of various ligand-receptor complexes. It appears to be due to the failure of hydrophobic ligands on dry filters, to be readily removed by aqueous media. A means whereby the vacuum can be held constant is described and shown to eliminate this artifact. PMID- 6285106 TI - Oxaprotiline: induction of central noradrenergic subsensitivity of its (+) enantiomer. AB - The effects of the tetracyclic antidepressant oxaprotiline and its two optically active enantiomers on the norepinephrine (NE) receptor coupled adenylate cyclase system were determined in slices of the rat cerebral cortex. While oxaprotiline does not change the response of the cyclic AMP generating system to NE after a single dose, chronic administration of the drug for 3 to 14 days down-regulates the receptor system. The noradrenergic subsensitivity is linked to a reduction in the Bmax value of beta-adrenergic receptors as assessed by (3H)-dihydroalprenolol binding without changes in the Kd value. The action of oxaprotiline on the NE receptor coupled adenylate cyclase system resides entirely in the (+)-enantiomer which is a potent inhibitor of the neuronal uptake of NE. The (-)-enantiomer of oxaprotiline which is a weak inhibitor of NE reuptake, failed, even in high doses, to modify the noradrenergic receptor system. Though not excluding co regulatory factors in addition to NE, the studies support the view that an enhanced and persistent NE receptor interaction is one of the prerequisites for the in vivo down-regulation of central noradrenergic receptor function. The results also suggest that the therapeutic activity of oxaprotiline may reside in its (+)-enantiomer. PMID- 6285107 TI - Cycloheximide: an adrenergic agent. AB - Cycloheximide, a widely used inhibitor of protein synthesis, stimulates glycogenolysis, gluconeogenesis and ureogenesis in isolated rat hepatocytes. The effects of cycloheximide were compared to those of norepinephrine. Both agents, cycloheximide and norepinephrine, produced slight increases in the levels of cyclic AMP (30% increases) which were blocked by propranolol. Interestingly, it was found that the metabolic actions of norepinephrine and cycloheximide (stimulation of glycogenolysis, gluconeogenesis and ureogenesis) were only slightly diminished by the beta adrenergic antagonist propranolol but abolished by the selective alpha 1 adrenergic antagonist prazosin. The ability of cycloheximide to inhibit protein synthesis was not affected by either prazosin or propranolol. It is concluded that the stimulation of glycogenolysis, gluconeogenesis and ureogenesis by cycloheximide in rat hepatocytes, is an effect of the antibiotic independent of its ability to inhibit protein synthesis and that is mediated through activation of alpha 1 adrenoceptors. The adrenergic activity of cycloheximide should be considered when this drug is used as an inhibitor of protein synthesis. PMID- 6285108 TI - Minireview: quantitative autoradiography of neurochemicals. AB - Several new methods have been developed that apply quantitative autoradiography to neurochemistry. These methods are derived from the 2-deoxyglucose (2DG) technique of Sokoloff (1), which uses quantitative autoradiography to measure the rate of glucose utilization in brain structures. The new methods allow the measurement of the rate of cerebral protein synthesis and the levels of particular neurotransmitter receptors by quantitative autoradiography. As with the 2DG method, the new techniques can measure molecular levels in micron-sized brain structures; and can be used in conjunction with computerized systems of image processing. It is possible that many neurochemical measurements could be made by computerized analysis of quantitative autoradiograms. PMID- 6285109 TI - Involvement of endogenous digitalis-like substance in genesis of deoxycorticosterone-salt hypertension. AB - In an attempt to evaluate the role of endogenous digitalis-like substance in the genesis of deoxycorticosterone (DOCA) -salt hypertension, the effects of intravenous anti-digoxin antibody on the blood pressure response were observed in male Wistar rats that underwent heminephrectomy followed by treatment with DOCA and saline. Administration of anti-digoxin antibody caused a marked decrease in the blood pressure which continued for about an hour. Such a change in the blood pressure was not observed in pertinent control animals. Thus, it seems that endogenous digitalis-like substance plays an important role in the genesis of DOCA-salt hypertension. PMID- 6285110 TI - Characterization and distribution of 3H-(3MeHis2)thyrotropin releasing hormone receptors in rat brain. AB - The characteristics and distribution of putative thyrotropin releasing hormone (TRH) receptors were studied in rat central nervous system using the TRH analogue 3H-(3MeHis2)TRH as a radiolabeled ligand. The analogue had a dissociation constant of 2.3 +/- 0.2 nM and a receptor density of 34 +/- 2 fm/mg protein in whole brain, homogenates. An association rate constant ot 1.6 x 10(-3) min-1nM-1 and a biphasic dissociation with rate constants of 2.6 x 10(-3) min-1 and 1.3 x 10(-4) min-1 were observed. The brain was dissected into ten regions, and detectable levels of binding were found in all regions. The highest levels were found in amygdala/piriform cortex area and the septal region, and the lowest levels were found in the cerebellar and cerebral cortex. Competition curves showed the methylated analogue to have approximately 7-fold higher affinity for the receptor than TRH. The higher affinity, along with lower nonspecific binding, accounts for the much improved sensitivity of the binding assay of the methylated analogue (70-80% specific binding) as compared to 3H-TRH (15-20% specific binding) and enables one to work with much lower tissue amounts. Use of the tritiated analogue will greatly aid in further studies of TRH receptors. PMID- 6285111 TI - Catechol estradiol induced implantation in the mouse. AB - Induction of implantation is among the most sensitive responses to estrogens. The ability of catechol estradiols, 4-hydroxy-estradiol-17 beta (4-OH-E2) and 2 hydroxy-estradiol-17 beta (2-OH-E2), to induce implantation in ovariectomized pregnant mice was compared to that of estradiol-17 beta. Delayed implantation was maintained by the daily administration of 2 mg of progesterone. A single injection of 3 ng of estradiol-17 beta, 50 ng of 4-OH-E2, or 2,000 ng of 2-OH-32 consistently induce a full complement of implantation sites in all animals. Before the estrogenicity of the latter steroid can be established the lack of contaminating estrogens must be proved. PMID- 6285112 TI - Naloxone does not affect pain sensitivity, mood or cognition in patients with high levels of beta-endorphin in plasma. PMID- 6285114 TI - The dimethylheptyl derivative of (-)-delta 8-tetrahydrocannabinol reduces the turnover rate of gamma-aminobutyric acid in the septum and nucleus accumbens. AB - Accumulating evidence suggests that the cannabinoids exert their action to reduce the turnover rate of acetylcholine in the hippocampus by an action in the septum via inhibitory gamma-butyric acid (GABA) containing interneurons. In the studies presented here administration of the potent dimethylheptyl derivative of (-) delta-tetrahydrocannabinol, which has previously been shown to reduce the turnover rate of acetylcholine in the hippocampus, reduces the turnover rate of GABA in the septum. A simple model in which cannabinoids transsynaptically activate inhibitory GABAergic septal neurons impinging on cholinergic septal neurons does not explain the data. A more complex model suggesting that inhibitory GABAergic septal interneurons innervate other inhibitory GABAergic septal interneurons has been hypothesized. PMID- 6285113 TI - An inhibitory role of beta-endorphin in central cardiovascular regulation. AB - The cardiovascular effects of beta-endorphin after administration directly into the nucleus tractus solitarii (NTS) of urethane anaesthetised rats were investigated. Unilateral injection resulted in a dose related fall in mean arterial pressure and heart rate. No change in respiratory frequency was prevented and the bradycardia reduced by pretreatment with locally applied naloxone (10 ng). This dose of the opiate antagonist had no effect on mean arterial pressure or heart rate when administered alone. Antiserum to beta endorphin (1:50 dilution) caused a rise in pressure and a tendency towards tachycardia on injection into the NTS, while it completely blocked the depressor response and bradycardia induced by beta-endorphin. These results are consistent with the view that a beta-endorphin-like peptide has a depressor role in the central nervous system. The hypotension may result from an effect within the central connections of the baroreceptor reflex arc, probably at the level of the NTS. PMID- 6285115 TI - Fluorescent antibody to membrane antigen (FAMA) procedure for determining susceptibility to varicella: an evaluation. PMID- 6285116 TI - A modified immunofluorescent test for detection of antibody against human cytomegalovirus. PMID- 6285117 TI - [Diagnosis of breast diseases using thermography and hyperbaric oxygenation]. PMID- 6285118 TI - [Radiographic and radionuclide signs of solid and soft fibromas of soft tissue]. PMID- 6285119 TI - [Efficacy of the new cephalosporin antibiotics cefotaxime, cefoperazone and lamoxactam]. PMID- 6285120 TI - [Mitochondrial mass in the rat myocardium and liver in chronic hypobaric hypoxia]. PMID- 6285121 TI - Epstein-Barr virus, clinical symptomatology and immunodeficiency in non-Hodgkin's lymphoma patients. PMID- 6285122 TI - [Effect of diatom particles on the development of a syngeneic leukemia in mice]. PMID- 6285123 TI - [Molecular biology of the foot-and-mouth disease virus]. PMID- 6285125 TI - Purification of rhodopsin on hydroxyapatite columns, detergent exchange, and recombination with phospholipids. PMID- 6285124 TI - Cooling enhances adenosine 3':5' monophosphate accumulation in thyrotropin stimulated dog thyroid slices. AB - Dog thyroid slices have been stimulated in vitro by thyrotrophin (TSH) at 37 degrees C and 25 degrees C. Adenosine 3':5'-monophosphate (cyclic AMP) accumulation was enhanced by cooling to 25 degrees C. This observation has been extended to kidney cortex slices stimulated by parathyroid hormone (PTH). However, this phenomenon is not general: it does not apply to thyroid slices stimulated by prostaglandin E1 (PGE1) or adrenal cortex slices stimulated by adrenocorticotrophic hormone (ACTH). Slight cooling provides a useful tool to influence biochemical mechanisms in intact cells and therefore the mechanism of action of cooling on cyclic AMP was investigated in dog thyroid slices stimulated by TSH. Adenylate cyclase and cyclic nucleotide phosphodiesterase activities as measured in acellular preparations decreased in parallel with the temperature. A decrease of the cyclic AMP efflux from the cell at 25 degrees C, although not measurable in this preparation, did not seem to be responsible for the phenomenon. computer simulation of the kinetics of the cyclic AP accumulation curve is compatible with the hypothesis of decrease in the desensitization rate of adenylate cyclase at 25 degrees C. This was demonstrated using an experimental protocol in intact slices and by measurements of adenylate cyclase activities in particulate preparations of slices pretreated or not by the hormone. This decreased adenylate cyclase desensitization can explain the higher cyclic AMP levels in TSH stimulated thyroid slices incubated at 25 degrees C. However, this does not exclude complementary mechanisms. PMID- 6285126 TI - Purification and characterization of a light-activated rod outer segment phosphodiesterase. PMID- 6285127 TI - Real time assay of rod disk membrane cGMP phosphodiesterase and its controller enzymes. PMID- 6285128 TI - Isolation and assay of a phosphodiesterase inhibitor from retinal rod outer segments. PMID- 6285129 TI - Measurement of retinal cGMP changes with quick-freeze techniques. PMID- 6285130 TI - Measurement of cyclic nucleotides in retina. PMID- 6285131 TI - Lipid-protein interactions: saturation transfer electron paramagnetic resonance of spin-labeled rhodopsin. PMID- 6285133 TI - Microdissection of retina and measurement of cyclic nucleotides. PMID- 6285132 TI - Isolation of cellular retinoid-binding proteins from bovine retina with bound endogenous ligands. PMID- 6285134 TI - Isolation of the major envelope glycoprotein of avian myeloblastosis virus. PMID- 6285135 TI - Membrane receptor for phosphomannosyl residues. PMID- 6285136 TI - 3-Deoxy-D-manno-octulosonate-8-phosphate (KDO-8-P) phosphatase. PMID- 6285137 TI - CTP:CMP-3-deoxy-D-manno-octulosonate cytidylyltransferase (CMP-KDO synthetase). PMID- 6285138 TI - Isolation of Escherichia coli structural genes coding for the glycogen biosynthetic enzymes. PMID- 6285139 TI - Enzyme-linked immunosorbent assay for antibodies to native and denatured DNA. PMID- 6285140 TI - Analysis of the crosslinking components in collagen and elastin. PMID- 6285141 TI - A bifunctional plasmid carrying the recA gene of E. coli. PMID- 6285143 TI - Influence of phage T3 and T7 gene functions on a type III(EcoP1) DNA restriction modification system in vivo. AB - The ocr+ gene function (gp 0.3) of bacteriophages T3 and T7 not only counteracts type I (EcoB, EcoK) but also type III restriction endonucleases (EcoP1). Despite the presence of recognition sites, phage DNA as well as simultaneously introduced plasmid DNA are protected by ocr+ expression against both the endonucleolytic and the methylating activities of the EcoP1 enzyme. Nevertheless, the EcoP1 protein causes the exclusion of T3 and T7 in P1-lysogenic cells, apparently by exerting a repressor-like effect on phage gene expression. T3 which induces an S adenosylmethionine hydrolase is less susceptible to the repressor effect of the SAM-stimulated EcoP1 enzyme. The abundance of EcoP1 recognition sites in the T7 genome is explained by their near identity with the T7 DNA primase recognition site. PMID- 6285142 TI - Read-through transcription from a derepressed Tn3 promoter affects ColE1 functions on a ColE1::Tn3 composite plasmid. AB - Mutations in the repressor encoded by the transposon Tn3 tnpR gene lead to increased levels of expression of two gene products: the mutant repressor (TnpR-) and the Tn3 encoded transposase, TnpA (Heffron et al. 1978; Chou et al. 1979a). Derivatives of the ColE1::Tn3 composite plasmid, RSF2124, with mutant Tn3 repressor exhibited the expected elevated levels of transposition. Unexpectedly, hosts containing these tnpR- derivatives produced enhanced levels of the ColE1 encoded toxin, colicin E1. The gene for colicin E1 maps far (0.23-0.98 MU) from the Tn3 insertion point (0.73 MU) (Fig. 1). The colicin E1 overproduction phenotype, designated Eop-, was complemented in trans by wild type repressor gene product (TnpR+) to the wild type phenotype, Eop+. Hosts with RSF2124 derivatives which expressed high levels of both mutant repressor and mutant transposase (TnpR , TnpA-) were Eop-. Hosts containing plasmids deleted for both tnpA and tnpR promoters were Eop+, while hosts with plasmids carrying a lac promoter substitution for the tnpA promoter were Eop-. These data support the idea that a cis-acting effect of increased transcription from the tnpA promoter into adjacent ColE1 DNA was the cause of colicin overproduction. Increased transcription activated a putative colicin augmentation function (caf) whose presence was required for the Eop- phenotype. Deletion mapping established that one boundary of the caf locus lies within 52 bases of the junction of the left end of Tn3 and ColE1 DNA. ColE1 DNA in this area contains an open reading frame which could encode either a 74 or a 63 residue protein (B. Polisky, unpublished DNA sequence data). The presence of increased levels of an mRNA transcript from this region and/or the increased expression of protein(s) from this transcript could result in an Eop- phenotype. Expression of the Eop- phenotype requires the presence of the host recE gene. Evidence is presented which suggests that the recA repressor, lexA protein, controls expression of the recE gene product, ExoVIII. PMID- 6285144 TI - Transcriptional termination sites in the b2 region of bacteriophage lambda that are unresponsive to antitermination. AB - A bacteriophage lambda cloning vector carrying the trp/lacW205 substitution is described. The vector facilitates the fusion in vitro of genetic control signals to the lacZ structural gene of Escherichia coli. This system was used to define transcriptional termination sites in the lambda b2 region. This region contains termination sites that are unresponsive to the lambda antiterminating proteins pQ and pN. PMID- 6285145 TI - Cloning of mglB, the structural gene for the galactose-binding protein of Salmonella typhimurium and Escherichia coli. AB - From libraries of EcoRI fragments of Salmonella typhimurium and Escherichia coli DNA in lambda gt7, phages could be isolated that carry mglB, the structural gene of the galactose-binding protein as well as other mgl genes. Lysogenization of an E. coli mutant carrying a defective galactose-binding protein with lambda gt7 mglB (Salmonella) restores full galactose transport and galactose chemotaxis. Both the E. coli mutant protein as well as the wild-type Salmonella galactose binding protein are synthesized in this strain. The EcoR1 fragments of both organisms carrying the mgl genes were 6 Kb long. They were subcloned into the multicopy plasmid pACYC184. The hybrid plasmid containing the Salmonella mgl DNA gives rise to the synthesis of large amounts of galactose-binding protein in the periplasm of E. coli. The protein can be precipitated by antibodies against the E. coli binding protein and is identical to the fully processed protein isolated from Salmonella typhimurium LT2. In vitro protein synthesis (Zubay-system) with either lambda gt7 mgl phages as well as the hybrid plasmid as DNA matrix produces the galactose-binding protein mainly in precursor form that is precipitable by specific antibodies. PMID- 6285146 TI - A secondary promoter for elongation factor Tu synthesis in the str ribosomal protein operon of Escherichia coli. AB - The str operon of Escherichia coli contains genes for ribosomal proteins S12 and S7 and for elongation factors EF-G and EF-Tu (Jaskunas et al. 1975). We have subcloned various segments of DNA from this operon onto multicopy plasmids. We found that cells carrying a recombinant plasmid which lacks the major promoter for the str operon but contains the 5' portion of the EF-Tu gene synthesize a novel protein which we have identified as a truncated EF-Tu molecule. Moreover, cells carrying plasmids with an intact EF-Tu gene synthesize the elongation factor at a 3- to 5-fold higher rate than haploid cells. Thus the EF-Tu gene can be expressed in the absence of the major promoter for the str operon. This expression is not due to read-through from plasmid promoters, but it is dependent on the presence of the distal portion of the EF-G gene on the plasmids. These results indicate that there is a secondary promoter for EF-Tu expression, apparently located within the structural gene for elongation factor EF-G. PMID- 6285147 TI - Lysis of Escherichia coli by induction of cloned phi X174 genes. AB - The largest of the fragments produced by AluI digestion of phi X174 RFI DNA comprises genes E and J as well as parts of genes D and F. This DNA fragment (1007 bp) was cloned into the lac z' gene of plasmid pUR222. In the recombinant plasmid pUH12, transcription of the phi X174 genes is controlled by the lac p-o region. Induction of the cloned genes by addition of the lac inducer, IPTG, resulted in lysis of the bacteria. Cloning of the corresponding AluI-fragment from phi X174am3 DNA, carrying an amber mutation in gene E, showed that the expression of this gene alone is sufficient to trigger cell lysis. The time interval between the addition of IPTG and the onset of lysis depended on the concentration of the inducer, however, the rate of lysis was similar at all IPTG concentrations used. PMID- 6285148 TI - Transformation of Streptococcus sanguis (Challis) by linear plasmid molecules. AB - The streptococcal erythromycin resistance plasmid pSM9 was used to study the problem of how the transforming activity of mixtures of two unique linear products of restriction enzyme digestion depends on the distance between the cleavage sites. In transformation of the Challis strain of S. sanguis, the transforming activity of mixed digests increased with increasing relative distances (x) between the restriction sites, where 0 less than or equal to X less than or equal to 0.5. To explain the experimental results, a mathematical model was proposed according to which the overall probability (p) of transformation resulting in a functional replicon is the product of the partial probabilities of initial single-strand pairing, circularization, and stability of the paired intermediate, all of which were assumed to depend on x. A linear relationship found between transformation frequency and p was taken to support the model. Transformation of Challis by mixtures of two linearized plasmid molecules with regions of internal nonhomology resulting in paired intermediates with insertion or substitution loops allowed either donor molecule to contribute to the transformation yield. PMID- 6285149 TI - Recombinant plasmids with genes for the biosynthesis of alkaline phosphatase of Escherichia coli. AB - Restriction maps of several recombinant plasmids representing a section of the E. coli K12 chromosome 35,000 bp in size with the genes phoA, proC and phoB were prepared. The orientation of phoA and the exact position of its N-terminal end on this map were determined by identifying a subfragment which carried the phoA promoter and by determining the nucleotide sequence of a 160 bp portion of this subfragment comprising the codons for the N-terminal end of pre-alkaline phosphatase. From this DNA sequence the leader sequence of alkaline phosphatase which consists of 21 amino acids was derived. PMID- 6285150 TI - The use of the plasmid pHA10 in the isolation of lambda PL promoter mutations. AB - The plasmid pHA10 provides a good positive selection system for PL promoter mutations. It contains the lambda EcoR1-D fragment cloned into the pBR322 plasmid. Inactivation of the temperature sensitive lambda repressor at 42 degrees C leads to transcription of the kil gene and host death. One group of survivors at 42 degrees C is saved due to mutations in the PL promoter. After several screening procedures one such mutant was isolated. Restriction enzyme analysis of the plasmid DNA followed by sequence determination showed it to be a 24 base pair deletion beginning half-way through the "Pribnow box" continuing through part of the -35 region. It removes the OL1 section of the OL operator plus one base pair of OL2. This mutation (called dpl) may be useful to the study of the lambda OLPL regulation system. The effect of base changes within the PL promoter on the promoter activity are discussed. PMID- 6285151 TI - Tn10 insertions directed in the pyr D - ser C region and improved mapping of pep N in Escherichia coli K12. PMID- 6285152 TI - Cloning of bacteriophage T5 promoters. AB - Bacteriophage T5 was subjected to combined hydrolysis with the restriction endonucleases PstI and HindIII and the resulting fragments were inserted into the plasmid pBR322. Selection of transformants for Aps-Tcr-phenotype made it possible to screen the hybrid plasmids that contained promoter sequences in the cloned fragments. Two PstI/HindIII fragments, 720 bp (51% of the T5 DNA length) and 1,200 bp (70%) were cloned in this study. Tcr levels for these plasmids were as high as 18 micrograms/ml and 75 micrograms/ml, respectively. The presence of Escherichia coli RNA polymerase binding sites on both fragments was shown using the nitrocellulose filter assay. These binding sites are situated between 35 bp and 95 bp from the HindIII cleavage site on the 1,200 bp fragment; and within 420 bp from the HindIII site on the 720 bp fragment. PMID- 6285153 TI - Endocrinological features and endometrial morphology in climacteric women receiving hormone replacement therapy. AB - Prolactin (PRL), follicle stimulating hormone (FSH), luteinizing hormone (LH), oestrone (E1), and oestradiol (E2) levels were determined in 204 women who were receiving hormone replacement therapy for their climacteric symptoms. The changes in these hormone levels and the endometrial morphology were studied in order to determine the effects of the replacement therapy. The women were divided into two groups: the first group of 120 women was treated with conjugated oestrogens administered cyclically, plus norethisterone acetate. The second group of 84 women received oral oestriol succinate, also administered cyclically but without additional progestogens. The oestrogen-progestogen therapy resulted in a disappearance of the climacteric symptoms and a significant decrease of FSH and LH levels. Oestriol therapy was less effective than the conjugated oestrogens as a replacement therapy. Oestriol therapy also resulted in a less remarkable decrease of gonadotrophin levels. There were no significant changes in prolactin levels in either group of women. The endometrial histology did not change significantly after either of the two hormone replacement therapies. PMID- 6285154 TI - An intrauterine progesterone contraceptive system (52 mg) used in pre- and peri menopausal patients with endometrial hyperplasia. AB - An intrauterine progesterone contraceptive system (IPCS) (52 mg) was inserted in 25 women with cystic endometrial hyperplasia. Among these women, 11 complained of heavy climacteric symptoms and also received an oestrogen replacement therapy consisting of conjugated oestrogens (0.625 mg/day) administered cyclically for 3 out of 4 wk. Prior to the therapy and after 6 mth of treatment, follicle stimulating hormone (FSH), luteinizing hormone (LH), prolactin (PRL), oestrone (E1) and oestradiol (E2) plasma levels were measured and endometrial histology was evaluated. In the women receiving IPCS treatment alone, there were no significant changes in FSH, LH, PRL, E1 and E2 plasma levels. However, there were remarkable changes found in their endometrial histology. In the remaining women receiving both treatments there was a sharp decrease in FSH and LH plasma levels and an increase in E1 and E2 plasma levels, while the prolactin levels remained unchanged. With the exception of two of these women, the endometrial histology changed remarkably. The endometrial morphology of the two exceptions remained unchanged. All climacteric symptoms disappeared after the administration of both IPCS and the oestrogen replacement therapy. The remarkable changes which did occur in the endometrial histology resulted in a less active glandular epithelium and stromal decidual formation, thus proving a useful effect of treatment. PMID- 6285156 TI - Modulation of benzodiazepine receptor binding by antiepileptic drugs and seizures. PMID- 6285155 TI - Reactivation of Epstein-Barr virus in pregnant women: social factors, and immune competence as determinants of lymphoproliferative diseases-a hypothesis. AB - In normal pregnant women, reactivation of Epstein-Barr virus (EBV) frequently occurs. Cellular immune responses are apparently suppressed, but high titer EBV specific antibodies of IgG class may compensate. The antibodies cross the placenta and protect the infant against primary infection for many months. Reactivation of EBV in pregnancy and oral excretion of virus by normal pregnant women could be one explanation why young children in large sibships (among families of low social classes) become infected early in life. Infants are likely infected by EBV from their pregnant mothers who shed virus in saliva. Moreover, the natural protection for several months in the perinatal period against Burkitt lymphoma and fatal EBV-induced lymphoproliferative diseases in congenitally immune deficient children is explained. The maturity and immunocompetence of the immune system at the time of primary infection by EBV and the size of the sibship seem to determine whether infectious mononucleosis occurs. Gammaglobulin derived from human cord blood may be a valuable source of viral-specific antibodies for serotherapy in immune deficient patients. PMID- 6285157 TI - Typing of herpes simplex virus strains by analysis of the early proteins. AB - In Herpes simplex virus (HSV) infected cells treated with the arginine analogue canavanine, early (alpha -) viral proteins accumulate and show a typical pattern in the acrylamide gel. This paper investigates the possibility of using this pattern for typing of HSV strains. Twenty strains were analyzed by this method and results were compared with typing by neutralization test. For 18 strains both methods agreed. Two strains regarded as intermediary strains by neutralization test could be clearly typed by the analysis of the early proteins. PMID- 6285158 TI - Epstein-barr virus VCA IgM and EBNA IgG antibodies titered by immunofluorescence in microplates. A semi-automated method based on microtiter system. AB - We describe methods of immunofluorescence in microplates for titration of EBV VCA IgM and EBNA-IgG antibodies and compare the sensitivity of the methods with glass slides and with microplates, showing the reproducibility of the methods. Results of VCA-IgM and -IgG, EA and EBNA titers obtained in four groups of patients are given, comprising cases of infectious mononucleosis, renal graft recipients, lymphoproliferative diseases and controls. PMID- 6285161 TI - Pulmonary cancer of triple cell differentiation with sarcoid reaction. Case report. PMID- 6285160 TI - Clinicopathological conference: secondary amenorrhea. PMID- 6285159 TI - [Glomangioma with muscular localization. Description of a case with peculiar histological characteristics]. PMID- 6285162 TI - Seroimmunity to polioviruses and B. pertussis. Population data for residents of Kansas City. PMID- 6285163 TI - Tuberculosis aggressively refractory to the triple drug therapy: case report. PMID- 6285164 TI - Conformational requirements of alpha 2-adrenergic receptors. AB - The conformationally restrained trans-extended and cis-folded isomers of 2-(3,4 dihydroxyphenyl)cyclobutylamine were used to establish the conformational requirements of presynaptic alpha 2-adrenergic receptors in the field-stimulated guinea pig ileum. The trans-extended isomer produced a concentration-dependent inhibition of the twitch response with an EC50 of 34 microM. The cis-folded analogue failed to produce a significant inhibition of the twitch response at concentrations up to 1 mM. These results suggest that the presynaptic alpha 2 adrenergic receptor prefers phenethylamines in the trans-extended conformation over the cis-folded conformation. The conformational requirements of alpha 1- and alpha 2-adrenergic receptors appear to be similar. PMID- 6285165 TI - Production of hydroxyl radical by decomposition of superoxide spin-trapped adducts. AB - The spin trapping of superoxide by nitrone spin traps, in particular 5,5-dimethyl 1-pyrroline-1-oxide (DMPO), was found to lead to the de novo production of hydroxyl radical and a nonradical species. Hydroxyl radical is then spin-trapped by DMPO, producing DMPO-OH. It was determined that a background level of approximately 3% DMPO-OH relative to DMPO-OOH is produced by this mechanism. Thus, the detection of hydroxyl radical by spin trapping with DMPO must be used with caution if the level of hydroxyl radical is less than 3% of the rate of superoxide generation. Several examples are cited in which investigators suggested that hydroxyl radical is produced when in fact the spin trapping of this free radical may have been an artifact of the system. PMID- 6285166 TI - [3H]Etorphine receptor binding in vivo. Small fractional occupancy elicits analgesia. PMID- 6285167 TI - Differential stereoselectivity of methotrimeprazine enantiomers for selected central nervous system receptor types. AB - Optical isomers of methotrimeprazine, an analgesic/neuroleptic, were investigated with respect to their ability to interact with six receptor types or subtypes. Bovine caudate nucleus tissue homogenates provided the dopamine, opiate, and serotonin receptor populations studied in these experiments. The radioligands used in saturation and binding competition experiments were tritiated dopamine, spiperone, dihydromorphine, 5-L-methionine enkephalin, naloxone, and 5 hydroxytryptamine. Saturation experiments verified acceptable performance of these in vitro receptor assay systems and indicated that a one-site binding model was adequate for each of these ligands under the experimental conditions employed. The competition experiments exhibited statistically significant (p less than 0.05) differences in isomeric effects only for dopamine and 5 hydroxytryptamine receptors. The more active isomer, levorotatory methotrimeprazine, was shown to be pharmacodynamically equivalent to chlorpromazine at these receptor types. When the magnitude of receptor stereoselectivity is plotted against an estimate of the more active isomer's affinity for that particular receptor, an excellent correlation is observed. This suggests that a high degree of stereoselectivity characterizes a highly specific drug/receptor interaction. These findings are compatible with the conclusion that methotrimeprazine does not produce analgesia via a direct action upon opiate receptors. PMID- 6285168 TI - Interactions of barbiturates of various pharmacological categories with benzodiazepine receptors. PMID- 6285170 TI - Structure of the saxitoxin binding site at sodium channels in nerve membranes. Exchange of tritium from bound toxin molecules. AB - The exchange of tritium into water from saxitoxin molecules that were radiolabeled at the C-11 methylene position was measured at 37 degrees in solution and in suspensions of brain membranes. High concentrations of membrane receptors were used to assure that more than 80% of the total saxitoxin (STX) present was specifically bound. The amount of back-exchanged tritium was determined either by measuring the radioactivity remaining in the STX, using a second binding assay, or by measuring the tritium in water using ion-exchange chromatography. The results show that the back-exchange is accelerated in the presence of the membranes, and that this is attributable solely to the nonspecific toxin binding. Little change in the back-exchange rate over that in solution occurs in specifically bound toxin molecules. These results place certain restrictions on the possible bonds and configurations of receptor-toxin complexes. PMID- 6285169 TI - Guanine nucleotide and cation regulation of the binding of [3H]cyclohexyladenosine and [3H]diethylphenylxanthine to adenosine A1 receptors in brain membranes. AB - Guanine nucleotides, divalent cations, and sodium differentially regulate agonist and antagonist binding to adenosine A1 receptors in brain membranes. Guanine nucleotides decrease the binding of the adenosine A1 receptor agonist [3H]N6 cyclohexyladenosine ([3H]CHA) to guinea pig and bovine brain membranes by about 50% at 1--3 microM, while not affecting binding of the antagonist [3H]1,3-diethyl 8-phenylxanthine ([3H]DPX) to A1 receptors in bovine brain. GTP decreases the potency of agonists competing for [3H]DPX binding by 3--6 times, without altering the potency of antagonists. This effect can be used to grade experimental substances along an adenosine agonist-antagonist continuum. The 66% inhibition of [3H]CHA binding by 1 mM EDTA, with no change in [3H]DPX binding, suggests that endogenous divalent cations may regulate adenosine receptor interactions. Removal of endogenous divalent cations by EDTA treatment greatly increases the enhancement of [3H]CHA binding by divalent cations. Specific binding of [3H]CHA to guinea pig brain is increased 150--170% by 0.3--1.0 mM Mn2+, Mg2+, and Ca2+ following EDTA preincubation, secondary to an increase in apparent affinity and receptor number. Sodium ions also selectively regulate the binding of [3H]CHA. Sodium decreases [3H]CHA binding 40%, whereas lithium and potassium are ineffective. Sodium does not affect [3H]DPX binding. PMID- 6285172 TI - Differential phosphorylation of (E)-5-(2-bromovinyl)-2'-deoxyuridine monophosphate by thymidylate kinases from herpes simplex viruses types 1 and 2 and varicella zoster virus. AB - 5-(2-Bromovinyl)-2'-deoxyuridine (BrVdUrd) is a potent and selective inhibitor of herpes simplex virus Type I (HS-I) and varicella zoster (VZ) virus replication but is much less potent against herpes simplex virus Type II (HS-II) replication. A possible enzymatic basis for this difference is reported here to involve the virus-coded dThd-dTMP kinases (EC 2.7.1.75) from the three virus strains. The thymidine kinases from the three virus strains were purified by affinity chromatography. In addition to catalyzing the phosphorylation of nucleosides, each of the three purified enzymes catalyzed the phosphorylation of thymidylate to its diphosphate but at strikingly different rates. The relative amounts of extractable virus-coded thymidylate kinases were estimated to be 100/2/40 for cells infected with HS-I, HS-II, and VZ viruses, respectively. Extracts of cells infected with HS-I virus catalyzed the phosphorylation of the monophosphate of BrVdUrd to its diphosphate. In contrast, the product was not detected with extracts from cells infected with HS-II virus. THe ratios of rates with 0.5 mM BrVdUrd monophosphate versus 0.1 mM dTMP as substrates for each of the purified dThd-dTMP kinases from HS-I, HS-II, and VZ viruses and the dTMP kinase from host cells were, respectively, 0.09, less than 0.002, 0.03, and less than 0.0002. These observations correlate with the relative sensitivities of these viruses to BrVdUrd in cell culture and suggest that, if BrVdUrd exerts its effect as a triphosphate, the inefficient phosphorylation of the monophosphate contributes to the insensitivity of HS-II virus to this agent. PMID- 6285171 TI - Actions of the histrionicotoxins at the ion channel of the nicotinic acetylcholine receptor and at the voltage-sensitive ion channels of muscle membranes. AB - Various histrionicotoxins tested on frog nerve-muscle preparations showed a qualitative family resemblance to one another. They blocked the nerve-evoked muscle twitch and depressed both the peak amplitudes and the decay time constants of end-plate currents. During repetitive stimulation they progressively decreased the rate of rise and prolonged the falling phase of muscle action potentials, the latter resulting, at least in part, from blockade of voltage-sensitive potassium channels. These results indicated that the histrionicotoxins act at three membrane channels: the channel associated with the acetylcholine receptor, the sodium channel, and the potassium channel. Closer study of perhydrohistrionicotoxin suggested either two topographically distinct sites of action at the acetylcholine receptor-ion channel complex, or one site and two ion channel complex conformations. One site or conformation only alters the kinetics of channel closure. As these sites become saturated, the end-plate current decay time constant asymptotically approaches a limiting value. The other site or conformation prevents the channel from opening altogether. Further analysis indicated that the binding site for perhydrohistrionicotoxin that alters the kinetics of channel closure has an affinity constant of 0.1 microM-1 at -90 mV and that this affinity may be sensitive to the membrane potential. The lipid protein interface is a suggested site of histrionicotoxin action, common to the three channels studied here as well as to other intrinsic membrane proteins affected by histrionicotoxins. PMID- 6285173 TI - Comparison of opiate inhibition of adenylate cyclase activity in neuroblastoma N18tG2 and neuroblastoma x glioma NG108-15 hybrid cell lines. PMID- 6285174 TI - Potentiation of opiate action in neuroblastoma N18TG2 cells by lipid incorporation. AB - The effect of cerebroside sulfate, phosphatidylserine, and other phospholipids on opiate receptor function in neuroblastoma N18TG2 cells was studied by incorporation of lipids into the membrane bilayer of viable cells. A concentration- and time-dependent incorporation of sulfatide by N18TG2 cells was observed. The incorporated lipid was not metabolized during the incubation period of up to 48 hr at 37 degrees. Optimal conditions for lipid incorporation were determined to be 4 days after the cell seeding and in 1% fetal calf serum. The incorporated lipid was established to be associated with the plasma membrane fraction of the crude cell homogenate. Furthermore, increases in Vmax but not Km values of the adenylate cyclase for Mg2+, ATP, and prostaglandin E1 were observed in neuroblastoma N18TG2 cells exposed to cerebroside sulfate for 4--6 hr. The incorporation of cerebroside sulfate or phosphatidylserine by N18TG2 cells did not increase the number of opiate binding sites in this cell line as determined by [3H]naloxone, [3H]etorphine, or 3H-labeled D-Ala2-Met5-enkephalinamide binding. Although there was an increase in the affinity of [3H]naloxone binding, linear correlation between the amount of cerebroside sulfate incorporated and the quantity of binding increase was not observed. However, augmentation of both the potencies and the efficacies (maximal inhibitory level) of morphine and enkephalin to regulate adenylate cyclase activity was observed after sulfatide incorporation. At the maximal concentration of cerebroside sulfate used (67 microM) the opiate receptor activity in N18TG2 cells approached that of NG108-15 cells. Identical treatment of N18TG2 cells with cerebroside or psychosine sulfate did not produce any potentiation of the opiate inhibition of adenylate cyclase. Of all of the phospholipids tested--phosphatidylserine, phosphatidylinositol, and phosphatidylcholine--only phosphatidylcholine produced a potentiation of the opiate effect. Both synthetic dipalmitoyl phosphatidylcholine or brain phosphatidylcholine could elicit the potentiation. PMID- 6285175 TI - [Expression of cloned genes in pro- and eukaryotic cells]. AB - Expression of cloned genes in new environment is reviewed. Gene expression is possible under control of their own regulatory elements in the cells of related organisms. Genes may also function in cells of taxonomically remote organisms; for example, the genes of lower eukaryotes are active in bacterial cells, the Drosophila gene -- in yeast cells. The main principles of construction of pro- and eukaryotic vectore capable to provide the expression of DNA sequences in corresponding recipient cells are discussed. PMID- 6285176 TI - [Integrase of lambda phage inhibits the activity of promotor Patt under in vivo and in vitro conditions]. AB - In vitro and in vitro transcription of the region of lambda phage DNA containing promoter Patt has been studied. Active transcription was observed in the vicinity of the att site and at the promoter Patt in vivo during the first minutes after infection. This transcription considerably decreases at the 1th minute, and stops completely at the 15th minute after infection. Maximum synthesis of integrase also takes place at the 15th minute after infection. In vitro the partially purified int protein (integrase) completely inhibited transcription from the promoter Patt. Hence, integrase is most probably a repressor of Patt in vivo and in vitro. PMID- 6285178 TI - [Cloning of the promoter of the RNA polymerase operon located before the structural gene of the beta-subunit]. AB - An EcoRI-fragment of the rpoBC operon carrying the end of the rplL gene, the intercistron region and the beginning of the rpoB gene cloned on the pBRH4 plasmid makes it tetracyclin-resistant, i. e. possesses the properties of a promoter. There is no appreciable difference in the degree of antibiotic resistance of recombinant plasmids carrying the main PJ promoter and the additional promoter. PMID- 6285177 TI - [Effect of nuclease S1 on DNA of the highly oncogenic simian adenovirus SA7(C8)]. AB - The effect at specific nuclease S1 on DNA and the complex viral DNA-terminal protein of the highly oncogenic simian adenovirus SA7(C8) was studied. It was shown that nuclease S1 did not digest the bound between DNA and terminal protein in the complex but residual amino acid(s) was cleaved out after digestion with pronase. The DNA obtained after nuclease S1 action could be ligated and its 5' ends were phosphorylated by polynucleotide kinase. PMID- 6285179 TI - [Nuclease S1 for DNA molecular weight determination in multiplasmid Escherichia coli strains]. AB - DNase treatment used to convert supercoiled DNAs to the open form is suitable for molecular weight determination by electron microscopy or agarose gel electrophoresis. The comparative experiments showed that S1-nuclease has advantages over DNase for molecular weight determination in preparations containing many species of plasmids of significantly differing sizes, isolated from multiplasmid E. coli strains. PMID- 6285180 TI - Structural studies of immunoglobulins spin-labeled at the carbohydrate moiety. PMID- 6285181 TI - Characterization of the activation of the human C1r complement molecule. AB - The proenzyme form of C1r was isolated by sequential chromatography from the euglobulin fraction of human serum on DEAE-Sepharose 6B-CL, CM-Sepharose 6B-CL and Sepharose S-300-CL. This C1r had the tendency to spontaneously activate within 60-90 min of incubation at 37 degrees C in presence of EDTA and more slowly in the presence of Ca2+. The spontaneous activation of C1r was found to be a bimolecular process and could be completely inhibited by DFP in the pH range 6 9 and in the presence of Ca2+ without affecting the hemolytic C1r activity. [14C]DFP bound to trace proteins in the 60-90 kD range, but not to C1r proenzyme. The spontaneous activation of C1r was diminished in the presence of EDTA by DFP, but could not be completely suppressed. EDTA acts by removing Ca2+ from C1r, thereby changing the conformation of the protein and causing an increased digestibility of the C1r H-chain. At temperatures above 0-4 degrees C this influence destroyed the ability of C1r proenzyme and enzyme to form macromolecular C1 and thereby abolished its hemolytic activity. We conclude from these results that the spontaneous C1r activation in the pH range 6-9 and in the presence of Ca2+ is due to contaminant proteases. C1r activated also spontaneously at higher pH values between pH 9 and 13.2, but the spontaneous activation ceased abruptly at pH 13.4. An intramolecular process of activation cannot be excluded at these high pH values. It is, however, not clear, whether this activation is a suitable model for the C1r activation in the C1 molecule, because the hemolytic activity of C1r was substantially diminished under the high pH conditions. PMID- 6285182 TI - Iprindole: a cornerstone in the neurobiological investigation of antidepressant treatments. PMID- 6285183 TI - Ontogeny and phylogeny of hormone receptors. PMID- 6285184 TI - [Arrhenoblastoma in 2 sisters. Report of 2 cases, coincidence with pregnancy in 1 patient]. PMID- 6285185 TI - [Effect of piracetam on cyclic adenosine monophosphate levels. Animal experiment study of the brain in the guinea pig]. PMID- 6285186 TI - Differences in murine procarcinogen activation enzymes are not accompanied by parallel differences in procarcinogen-induced sister-chromatid exchange. AB - C57Bl/6 and DBA/2 mice, strains in which there is marked induction of hepatic monooxygenase activity by phenobarbital, were tested for in vivo sister-chromatid exchange (SCE) formation in response to cyclophosphamide, an agent metabolized by this inducible enzyme system. Baseline SCE frequencies were between 4 and 6 SCEs/cell in regenerating liver and bone marrow of both strains of mice. Administration of cyclophosphamide (5mg/kg) led to an increase of nearly 8 SCEs/cell in both tissues of C57Bl/6 mice and an increase of more than 10 SCEs/cell in DBA/2 mice. Prior exposure to phenobarbital induced p chloromethylaniline demethylase activity in regenerating liver of both mouse strains approx. 6-fold, but the changes in measured SCE frequencies were not significantly different from those obtained in the absence of enzyme induction. These results, together with our previous observation that induction by 3 methylcholanthrene of benzo[a]pyrene hydroxylase activity in the same mouse strains was not accompanied by a comparable change in benzo[a]pyrene-induced SCE formation, reinforce the impression that simple assays of differences in mixed function oxidase activities may not necessarily be good predictors of hereditary differences in the response to genetic damage by procarcinogens which are presumed to be metabolized by these enzymes. PMID- 6285187 TI - Lack of association between induction of dominant-lethal mutations and induction of heritable translocations with benzo[a]pyrene in postmeiotic germ cells of male mice. AB - Benzo[a]pyrene was tested for induction of dominant-lethal mutations in germ cells of male mice. Clear-cut dominant-lethal effects were induced in middle and early spermatozoa. In contrast to the dominant-lethal observed the study showed no detectable increase in heritable translocations for these stages over the spontaneous level. Thus, the results provide another example of a chemical mutagen that is effective in inducing dominant-lethal mutations but relatively ineffective in inducing heritable translocations in male postmeiotic germ cells. PMID- 6285188 TI - Microgravimetric analysis of nerve edema. AB - A technique for microgravimetric analysis of nerve edema was used to demonstrate increased water content in hexachlorophene neuropathy. The method was modified from one used previously to quantify brain edema by using density gradient columns prepared with two nonaqueous fluids of high and low specific gravity. This method revealed a wide separation between hexachlorophene-treated and control nerves. Gradient position was then related to specific gravity, which averaged 1.03426 in hexachlorophene-treated nerves versus 1.04921 for control nerves. This, in turn, was correlated with the percentage of water in the tissue. Water content of hexachlorophene-intoxicated nerves was approximately 10% greater than control nerves. A major advantage of this technique is its sensitivity in detecting edema in small tissue samples. PMID- 6285189 TI - Superficial mycoses: ketoconazole treatment. PMID- 6285190 TI - Enzymes of purine and pyrimidine metabolism from the human malaria parasite, Plasmodium falciparum. AB - Plasmodium falciparum trophozoites were isolated by mechanical rupture of infected human erythrocytes followed by a series of differential centrifugation steps. After lysis with sonication, the 100 000 x g supernatant of parasites and uninfected host cells was used to determine the specific activities of a number of enzymes involved in purine and pyrimidine metabolism. P. falciparum possessed the purine salvage enzymes: adenosine deaminase, purine nucleoside phosphorylase, hypoxanthine-guanine phosphoribosyltransferase (PRTase), xanthine PRTase, adenine PRTase, adenosine kinase. The last two enzymes, however, were present at much lower activity levels. Hypoxanthine was converted (presumably via IMP) into adenine and guanine nucleotides only in the presence both of supernatant and membrane fractions of P. falciparum. Two enzymes involved in the de novo synthesis of pyrimidines, orotic acid PRTase, and orotidine 5'-phosphate decarboxylase, were present in parasite extracts as were the enzymes for pyrimidine nucleotide phosphorylation: UMP-CMP kinase, dTMP kinase, nucleoside diphosphate kinase. Xanthine oxidase, CTP synthetase, cytidine deaminase and several kinases for the salvage of pyrimidine nucleosides were not detected in the parasites. Both phosphoribosyl pyrophosphate synthetase and uracil PRTase were present but at low activity levels. Human erythrocytes displayed similar but not identical enzyme patterns. Enzyme specific activities, however, were generally much lower than those of the corresponding parasite enzymes. PMID- 6285191 TI - Purification, characterization and inhibition by MK-401 of Fasciola hepatica phosphoglyceromutase. AB - Phosphoglyceromutase (EC 2.7.5.3) of Fasciola hepatica was purified 1390-fold to homogeneity. The enzyme had a molecular weight of 120 000 and was a tetramer composed of identical 30 000 molecular weight subunits. The enzyme was 2,3 dephosphoglyceric acid dependent, possessed reactive sulfhydryl groups and was inhibited irreversibly by iodoacetamide, and N-ethylmaleimide and reversibly by p chloromercuribenzoate and 5,5'-dithiobis(2-nitrobenzoic acid). Initial velocity studies suggest that reaction occurred via a sequential mechanism and that MK-401 was a competitive inhibitor versus both 3-phosphoglyceric acid and 2,3 diphosphoglyceric acid. PMID- 6285192 TI - Cytomegalovirus infection in day-care center. PMID- 6285193 TI - Biliary atresia and reovirus type 3 infection. PMID- 6285194 TI - The NK cell: a phagocyte is lymphocyte's clothing? PMID- 6285195 TI - Protein folding by numbers. PMID- 6285196 TI - Lysosome proton pump identified. PMID- 6285197 TI - Expression of murine H-2Kb histocompatibility antigen in cells transformed with cloned H-2 genes. AB - Cosmids containing H-2 histocompatibility antigen genes of the H-2b haplotype have been isolated. One of these genes expresses a 45,000 molecular weight protein, indistinguishable from H-2Kb when introduced into mouse L cells. These H 2Kb transformed L cells can be killed by allospecific anti-H-2Kb cytotoxic T cells. Moreover, when infected with influenza virus, they can be killed by an H 2Kb-restricted, influenza virus-specific cytotoxic T cell line. These results show that expression of the H-2Kb gene product on the L-cell surface is sufficient to make it a target for specific T-cell killing. PMID- 6285198 TI - Adenosine-induced slow ionic currents in the Xenopus oocyte. AB - Adenosine and its 5'-phosphorylated congeners evoke specific membrane-mediated responses in excitable tissues. Available data suggest that inhibition of the target cell occurs due to hyperpolarization, and in some preparations a compound effect of ATP (excitation and inhibition) has been found. However, the ionic mechanism of the purinergic-mediated response has not been studied by standard intracellular voltage-clamping techniques. Recently, we have discovered purinergic receptors in the Xenopus oocyte, a well defined giant cell amenable to rigorous electrophysiological and biochemical studies. We report here that in these cells, adenosine-induced slow membrane responses consisted of an early depolarizing (D) transient current carried by Cl ions, followed by a steady hyperpolarizing (H) current involving K+ ions. The relative potency sequence for the D current was ATP congruent to ADP greater than AMP congruent to adenosine; this order was reversed for the H current. PMID- 6285199 TI - Injection of subunits of cyclic AMP-dependent protein kinase into cardiac myocytes modulates Ca2+ current. AB - beta-Adrenergic stimulation of the heart is thought to increase cardiac muscle contractility by activation of cyclic AMP-dependent protein kinase and concomitant increase in the phosphorylation of certain proteins (for refs see refs 1-6). Electrophysiological studies have shown that the stimulation of cardiac beta-adrenoreceptors, the external application of cyclic AMP or its analogues to Purkinje fibres, or the injection of cyclic AMP into single myocytes can increase the slow inward current (Isi) during the plateau phase of the action potential (AP). In heart muscle this current is mainly carried by Ca2+ (refs 10, 11) and it has been suggested that cyclic AMP-dependent phosphorylation of some component of the calcium channel increases the amount of Ca2+ which enters the cell during depolarization. We have investigated this hypothesis by examining the electrical responses of isolated guinea pig ventricular myocytes to pressure injections of subunits of the cyclic AMP-dependent protein kinase. We report here that injection of the catalytic subunit (C) resulted in a lengthening of the action potential duration (APD) and an increase in the height of the plateau as well as the amplitude of Isi. By contrast, the injection of regulatory subunit (R) shortened the APD of fast and slow response APs, an effect which was reversed by adrenaline. PMID- 6285200 TI - The DNase I sensitivity of Xenopus laevis genes transcribed by RNA polymerase III. AB - Since the initial discovery that the DNase I sensitivity of the globin genes in different cell types correlates with globin gene expression, this relationship has been shown to hold true for a variety of genes, including the genes for ovalbumin, conalbumun, alpha- and beta-globin in chicken, several heat-shock proteins in Drosophila, the r-chromatin of Tetrahymena and the viral polyoma minichromosome. Although genes transcribed by RNA polymerases I and II have been studied extensively, the genes transcribed by RNA polymerase III have not. We have therefore investigated the DNase I sensitivity of transfer RNA (tRNA) and oogenetic 5S RNA genes in the liver and erythocyte nuclei of Xenopus laevis. The oogenetic 5S genes are not transcribed in any known somatic cell, and tRNA genes are transcribed in the hepatocyte but are inactive in the erythrocyte. We show here that, although in these two cell types the correspondence between DNase I sensitivity and gene transcription holds good for globin and the ribosomal genes, the tRNA and oogenetic 5S genes are DNase I sensitive in both liver and erythrocyte nuclei. Thus for the genes transcribed by polymerase III the correspondence of sensitivity and expression breaks down. PMID- 6285201 TI - Is avian adeno-associated virus an endogenous virus of chicken cells? AB - The adeno-associated viruses (AAV) are defective parvoviruses which produce infective progeny only in cells co-infected with a 'helper' adenovirus (Ad). Both human and simian AAV have been recovered from human and simian primary cell cultures following their inoculation with 'AAV-free' Ad. Whereas some studies have suggested that AAV exists in a latent state in these cells, others have indicated that the AAV genome is capable of establishing and maintaining a latent state in defined laboratory conditions which mimic the situation proposed for the 'latent' AAV recovered from human and simian tissues. Here, avian adeno associated virus (AAAV) was consistently recovered from limiting dilutions of purified and unpurified avian Ad stocks propagated in embryonating chicken eggs derived from two independently raised flocks of White Leghorn (WL) chickens but not when these Ad stocks were propagated in duck cells. These observations suggest that AAAV is a latent endogenous virus of at least some flocks of WL chickens. PMID- 6285202 TI - Warts and all. PMID- 6285203 TI - De novo methylation and expression of retroviral genomes during mouse embryogenesis. AB - Retrovirus genomes introduced into mouse zygotes by microinjection of cloned DNA, or into morula stage pre-implantation mouse embryos by infection with Moloney murine leukaemia virus (M-MuLV), became de novo methylated and were blocked in expression. No restriction of virus expression and no de novo methylation were observed when post-implantation mouse embryos were infected with virus. Efficient de novo methylation activity may be an important characteristic of gene regulation in early mouse embryos. PMID- 6285204 TI - Single base substitution in an intron of oxidase gene compensates splicing defects of the cytochrome b gene. AB - An extragenic suppressor mutation, mim2-1, which compensates yeast mitochondrial mutants deficient in splicing of the cytochrome b gene, has been mapped and sequenced. The mutation is due to a single G leads to A transition in the long open reading frame of the fourth intron of the oxidase subunit one gene. It causes the replacement of a glutamic codon by a lysine codon and the expression of a novel mRNA maturase active in splicing. Evolution and regulatory connections between homologous introns of nonhomologous genes are discussed. PMID- 6285205 TI - Glycine receptor alteration in the mutant mouse spastic. PMID- 6285208 TI - Maintenance of broad host range plasmid RK2 replicons in Pseudomonas aeruginosa. PMID- 6285206 TI - Origin of lignans in mammals and identification of a precursor from plants. PMID- 6285207 TI - Insertion of diphtheria toxin into and across membranes: role of phosphoinositide asymmetry. PMID- 6285209 TI - Amplification of endogenous myc-related DNA sequences in a human myeloid leukaemia cell line. PMID- 6285210 TI - [Endocrine pancreatic tumors]. PMID- 6285211 TI - [Chronic progressive external ophthalmoplegia (author's transl)]. PMID- 6285212 TI - The glucagonoma syndrome. PMID- 6285213 TI - Major determinants of plasma aldosterone levels in chronic uremia on dialytic treatment. PMID- 6285214 TI - [Glioblastoma multiforme developing after a gunshot injury of the brain (author's transl)]. AB - We report the case of a 28-year-old male who suffered a frontal penetrating gunshot injury with subsequent bifrontal brain abscess and subdural empyema, and five years later developed a large bifrontal tumour at the precise site of the meningo-cerebral scar and posttraumatic defect. Histological examination showed a glioblastoma multiforme adjacent to the dural scar and, in addition, old suture material was found within the glioma tissue. In spite of combined radiation and polychemotherapy the patient died eleven months after partial tumour resection. The temporal and local association of missile injury with subsequent recurred abscess and scar formation and the malignant glioma is highly suggestive of a causal relationship between trauma and the development of a brain tumour. PMID- 6285215 TI - [An anatomical and technical note for neurosurgery of the jugular foramen tumor [author's transl)]. AB - The basic anatomy of the jugular foramen, some diagnostic principles of the jugular foramen tumors with presentation of our experiences of the 13 cases (6 neurinomas, 6 chemodectomas and 1 meningioma), and the detailed surgical technique used in their removal and its result are described. According to the extension of the tumor, one of the following operative approaches can be selected. A) Transjugular approach consists of a retromastoideal craniectomy following a radical mastoidectomy. The posterior wall of the jugular foramen is scraped out and the sigmoid sinus and the internal jugular vein are resected with the tumor either confined in the jugular foramen or extending out of the skull. B) Transjugular-transpetrosal approach is suitable for the tumor in the jugular foramen extending out of the skull as well as into the CP angle. The petrosal bone is more scraped out for removal of the tumor extending into the CP angle than in the above transjugular approach. In the case of the tumor extending into the CP angle and the upper clivus, the upper petrosal bone medial to the C-5-C-6 segments of the internal carotid artery are also removed and an extensive exposure of the intracranial tumor is obtained. C) A suboccipital craniectomy with retrolabyrinthine removal of the petrosal bone is suitable for large glosso pharyngeal neurinomas extending into the CP angle but not out of the floor of the skull. In such cases, after removal of the CP angle tumor in the usual way, resection of the remaining jugular foramen tumor is easily performed through opening the petrosal bone forming the medial portion of the dome of the jugular fossa. PMID- 6285217 TI - Familial gliomas: report of four families, with chromosome studies. AB - Four families having multiple family members with cerebral gliomas are presented. Genetic studies were done in some, but no chromosomal abnormalities were found in this group of patients or their families. The authors recommend that careful attention be given to the family history of all glioma patients and that more extensive genetic studies be done. The formation of a registry to report cases of familial gliomas is also suggested. PMID- 6285216 TI - Melatonin does not affect luteinizing hormone-releasing hormone binding to neonatal rat anterior pituitary membranes. PMID- 6285218 TI - Long term survival among patients with malignant brain tumors. AB - Eight of 57 patients (14%) with malignant astrocytomas lived at least 2 years. The mean survival time was 143 weeks (range, 104 to 250 weeks). All of the patients were treated with operation, radiation, and chemotherapy. Four of the 8 patients died because of tumor recurrence, 1 died from a second primary tumor, 2 died of cases unrelated to the tumor, and 1 is still alive. Diffuse cortical dysfunction associated with cortical atrophy that could not be related to tumor regrowth and was not explained by focal deficits, psychotic of depressive thought disorders, metabolic or endocrine abnormalities, or hydrocephalus developed in the 3 longest-surviving patients. The diffuse dysfunction was initially apparent only through psychometric testing, but eventually became so disabling as to result in 2 of the 3 patients retiring from work. Although small, but gratifying, gains have been made in the treatment of patients with malignant brain tumors, accompanying these gains have been problems of a magnitude that is only now beginning to be appreciated. PMID- 6285219 TI - Effect of age and reoperation on survival in the combined modality treatment of malignant astrocytoma. AB - Before the advent of the operating microscope and the carbon dioxide laser, reoperations were performed in less than 5% of patients with malignant astrocytoma. Between 1978 and 1981, a consecutive series of 74 adult patients were prospectively treated therapy, all patients eligible for further treatment were offered reoperation with the microscope and/or laser before treatment with Phase I agents (microwave hyperthermia, dimethyl sulfoxide (DMSO)-Adriamycin, DMSO-Cytoxan, or azaridinylbenzoquinone (AZQ)). Forty-six per cent of the patients were referred from outside institutions for intensive treatment. In 36 months, 40 patients received second operations directed at their tumor and had a median calculated survival from the time of reoperation of 37 weeks. The length of survival after the second operation was independent of patient age, performance status, tumor grade, and interoperative interval. Sixty-five per cent of patients under 40 and 48% of patients over 40 underwent reoperation with minimal morbidity and no deaths. The single most important prognostic factor for survival in both reoperated and single-operated patients was age. Tumor grade had no influence on survival in the series as a whole, in patients under 40 years of age, or in reoperated patients. The calculated median survival for the entire series was 15 months, with a predicted 2-year survival rate of 0.25. These figures include all patients treated without exclusion for incomplete radiotherapy or chemotherapy. It is concluded that reoperation for malignant astrocytoma is safe, feasible, and of potential benefit in combination with other therapies. The routine use of reoperation to "set up" other treatment modalities deserves further study. PMID- 6285220 TI - Impaired immunocompetence in patients with malignant gliomas: the possible role of Tg-lymphocyte subpopulations. AB - T-lymphocyte subpopulations (Tg or "suppressor cells" and Tm or "helper cells") in the peripheral blood of 16 patients with malignant intracranial gliomas were analyzed. The number of Tm-lymphocytes was very close to normal in both pre- and postoperative samples, whereas Tg-lymphocytes were significantly increased preoperatively (18.55 +/- 1.11 vs. 12.72 +/- 1.19 p less than 0.001) and were still slightly higher than normal after operation and radiation treatment. These preliminary findings suggest a possible role of T-suppressor lymphocytes in the impairment of cell-mediated immunocompetence that has been observed repeatedly in patients with malignant gliomas. PMID- 6285221 TI - Ventricular catheter in the cistern of the traverse fissure: a cause of shunt malfunction. PMID- 6285222 TI - Role of viruses in the induction of primary intracranial tumors. AB - The role of viruses in the induction of primary intracranial tumors is reviewed. Papovaviruses of the simian virus 40 (SV40) group are used as a representative model, and a distillation of the literature on virus induction of intracranial tumors in laboratory animals and neoplastic transformation of cells in culture is presented. The molecular sequence of events during tumor induction and neoplastic cell transformation is also discussed. Recent evidence that the papovaviruses play a role in the formation of human brain tumors is summarized. PMID- 6285223 TI - Selective inhibition of 'motor endplate-specific' acetylcholinesterase by beta endorphin and related peptides. AB - The effects of beta-endorphin (lipotropin 61-91) and related naturally-occurring peptides upon acetylcholinesterase activity in rat hind-limb muscles was investigated. beta-endorphin weakly inhibited the activity in a plasma membrane enriched fraction. The inhibition by beta-endorphin of the membrane-associated acetylcholinesterase was less marked when the fractions were prepared from muscles which had been denervated 4-6 days previously. The membrane-associated acetylcholinesterase was solubilised from normal muscle preparations and separated by sucrose density gradient centrifugation into three major peaks (16S, 10S and 4S). beta-Endorphin inhibited the activity in the 16S peak but not that in the 10S and 4S peaks, whilst tensilon, a competitive inhibitor of acetylcholinesterase, inhibited the activity of all three peaks. beta-Endorphin inhibited the activity in the 16S peak but not that in the 10S and 4S peaks, whilst tensilon, a competitive inhibitor of acetylcholinesterase, inhibited the activity of all three peaks. beta-Endorphin inhibited the 16S activity in a concentration-dependent manner and its action was partly prevented if naloxone was added simultaneously. Purified natural porcine and bovine beta-endorphin were equipotent in terms of effective concentration range but the maximum inhibition was greater with the bovine peptide. beta-Lipotropin was approximately 4 times less potent than beta-endorphin, whilst C-fragment (lipotropin 61-87) was 100 times less potent. Prolonged treatment with collagenase did not reduce the catalytic activity of 16S acetylcholinesterase, but it was no longer susceptible to the inhibitory action of beta-endorphin. Kinetic studies indicated a complex type of inhibition by beta-endorphin (hyperbolic Lineweaver-Burke plot). Methionine enkephalin inhibited acetylcholinesterase in a weakly non-competitive manner and its action was not abolished if the enzyme was predigested with collagenase. beta-Endorphin produces a novel form of inhibition of acetylcholinesterase, acting only on the 16S (A12 or 'motor endplate-specific') form of the enzyme. The findings are discussed in the light of evidence that beta endorphin-related immunoreactivity is expressed in motor nerve axons in the immature rat. PMID- 6285224 TI - Signal transduction across cellular membranes. PMID- 6285225 TI - Immunocytochemical studies of human peripheral nerve with serum from patients with polyneuropathy and paraproteinemia. AB - Immunohistochemical binding of IgM paraproteins to nerve was studied using the immunoperoxidase technique with serum from 10 patients with benign plasma cell dyscrasia and neuropathy. We stained the myelin sheaths of peripheral nerves and roots fo five patients who had myelin-absorbable IgM paraproteins. Two patients with IgM paraproteins that did not react with myelin showed predominant staining of axons, while three were completely negative. Serum specimens from normal volunteers and patients with paraproteinemias or ALS were also unreactive. Immunocytochemical methods can detect IgM paraproteins with an affinity for nerve antigens and may assist in the diagnosis and classification of plasma cell dyscrasia associated neuropathy. PMID- 6285226 TI - Infantile glycogen storage myopathy in a girl with phosphorylase kinase deficiency. AB - A 19-month-old girl with moderate hypotonia was studied. Histochemical and electronmicroscopic findings revealed that many skeletal muscle fibers contained an excess amount of glycogen. The phosphorylase reaction was normalized only after activation with 5' AMP. Biochemical studies showed an increased glycogen content and decreased activities of phosphorylase "a" and an active form of phosphorylase kinase, whereas activities of total phosphorylase, total phosphorylase kinase, and cyclic AMP-dependent protein kinase were all in the normal range. Thus, phosphorylase kinase in the patient's muscle seemed to be a variant form, which was activated partially under the physiologic condition. This condition may be inherited as an X-linked recessive trait. PMID- 6285227 TI - Vasculitic neuropathy in rheumatoid disease and Sjogren syndrome. AB - Two patients with rheumatoid arthritis and one with Sjogren syndrome had a severe sensorimotor neuropathy preceding or up to 5 years after the onset of the disease. Electrophysiologic and sural nerve biopsy studies revealed an axonal neuropathy. Myelinated fibers were affected to a greater extent than unmyelinated axons. Peripheral nerve damage was related to occlusion of the vasa nervorum, since vasculitic involvement of epineurial vessels was observed in all patients. Despite the severity of the neuropathy, it may recover, because, compared with axons, Schwann cells are perhaps less vulnerable to ischemia. PMID- 6285228 TI - Cytochrome-c-oxidase deficiency in a floppy infant. PMID- 6285229 TI - [Update on antibiotic therapy. 7) Cefotiam]. PMID- 6285230 TI - Acute membrane responses to viral action. AB - The effects of Sendai, a paramyxovirus, on the functional activity of 3 cell types, have been studied in vitro to establish whether a virus alone can cause pathophysiological changes. Neuronal cells are depolarized and suffer a loss of excitability which was attributed to an increase in membrane conductance. Spontaneously beating cardiac cells initially stop beating and then beat more rapidly and asynchronously. Anterior pituitary cells release hormones. In all 3 cases the effects are transient and the cells recover completely. PMID- 6285231 TI - Selective destruction of the ventral noradrenergic bundle but not of the locus coeruleus elevates plasma levels of beta-endorphin immunoreactivity in rats. AB - The present study examines the role of brain networks of noradrenaline (NA) in the control of pituitary secretion of beta-endorphin into systemic plasma. The blocker of NA synthesis, FLA-63, elicited a pronounced elevation in levels of beta-endorphin immunoreactivity (beta-EI) in systemic plasma and depleted the anterior but not the neurointermediate lobe content of beta-EI. This finding suggests the existence of a noradrenergic mechanism tonically inhibiting the secretion of adenohypophyseal pools of beta-EI into plasma. Selective destruction of the ventral noradrenergic bundle diminished the NA content of the hypothalamus but not the cortex, and produced an increase in plasma levels of beta-EI. Lesions of the locus coeruleus, the primary origin of the dorsal noradrenergic bundle, in contrast, decreased NA levels in the cortex but not the hypothalamus and failed to modify beta-EI levels in plasma. the ventral bundle may, thus, via a hypothalamic site of action, be the noradrenergic pathway inhibiting the secretion of beta-EI into the circulation. PMID- 6285232 TI - Nutrition classics. The Journal of Nutrition. Volume 24, 1942. Pages 213-224. The effectiveness of linoleic, arachidonic, and linolenic acids in reproduction and lactation. F.W. Quackenbush, F.A. Kummerow and H. Steenbock. PMID- 6285233 TI - Gastrointestinal factors in hunger and satiety. AB - The evidence for the participation of signals from the gastrointestinal tract in hunger and satiety is reviewed. Various methods of subjecting only certain portions of the gastrointestinal tract to food are described including sham feeding, crossing of the intestines of two animals, and intragastric, intraduodenal and intravenous feeding. These methods have revealed that animals eat more when only the mouth is exposed to food, but consume less when food enters the stomach directly and still less when feeding themselves intravenously. The role of the stomach in satiety is discussed in relation to the results of the many experiments in which intragastric loads were administered. These experiments generally revealed that gastric loads suppressed intake in proportion to their caloric value, but that compensation was not precise. A similar analysis is made of the role of the intestines in satiety. These studies present evidence for osmoreceptors and glucoreceptors, distention, and pressure, all of which may participate in satiety. The numerous effects of vagotomy on food intake are reviewed and an attempt is made to separate afferent from efferent effects and vagal effects from nonspecific surgical injury. The role of other afferents is explored. A brief survey of the most recent studies on humoral factors indicates that cholecystokinin is probably acting as a satiety agent, but its mode of action remains unknown. The changes in the gastrointestinal tract that accompany an increase or decrease in food intake reveal adaptive changes that occur in the organ. The evidence for gastrointestinal factors in satiety of the suckling animal is presented. Finally, a hypothesis is presented in which the various factors that bring a meal to an end are included. Preabsorptive satiety may consist of multiple factors which probably include signals from osmoreceptors and possibly other chemoreceptors in the upper small intestine. The signals from these receptors are neural while other products of digestion stimulate the release of humoral agents, such as cholecystokinin, which may act locally, in the liver, or on the central nervous system. Stretch or distention of the intestines at various locations may also contribute to preabsorptive satiety. PMID- 6285234 TI - Habituation and sensitization in an aneural cell: some comparative and theoretical considerations. AB - Several shorter- and longer-term non-associative modifications in behavior are known to occur in neural as well as aneural systems. Thus neural investment is not essential for these phenomena to occur. Cellular studies of these behaviors in Protozoa, where a single cell is also a whole organism, may be useful in investigating the evolution of mechanisms underlying these plastic behavioral changes. PMID- 6285235 TI - The role of norepinephrine in feeding behavior. AB - When dopamine-beta-hydroxylase is inhibited with FLA-63 (10 mg/kg) free feeding behavior is disrupted in satiated rats. While the average number of meals taken was not different from vehicle injected controls, meal size was decreased 58% in the first 9 hr after treatment with FLA-63. In starved animals, FLA-63, when given alone, produced little effect on feeding behavior, even though norepinephrine depletion was in excess of 40%. When given in combination with RO4 1284 (5 mg/kg), a vesicular reuptake inhibitor, feeding was reduced to 16% of control intake and norepinephrine was specifically depleted 99%. Feeding was reliably reinstated in animals which received FLA-63 plus RO4-1284 with either dl threo-DOPs, a metabolic precursor to NE, or direct intrahypothalamic injections of NE. These findings suggest that the feeding inhibition observed after treatment with FLA-63 plus RO4-1284 is due to disruption of transmission in brain NE systems. A non-anorectic dosage of L110-140 (3.73 mg/kg), a specific FLA-63. Taken collectively, these findings suggest that the primary role of NE in feeding is maintenance of the consummatory response and that these effects are expressed in relation to activity in other neurochemical systems. PMID- 6285236 TI - Vasopressin: a homeostatic effector in the febrile process. AB - This review compares the physiological changes which accompany infection and fever with the effects of the peptide, arginine vasopressin (AVP). AVP may act as a neuromodulator, a releasing factor, or a hormone to induce responses which are opposite to those homeostatic changes accompanying fever. Since AVP is released into blood and brain during fever, it is hypothesized that AVP contributes to the maintenance of homeostasis in the infected organism. PMID- 6285237 TI - Cefotiam versus cefoxitin for treatment of severe or complicated urinary infections. PMID- 6285238 TI - [Virological studies of feces and sewage in Berlin]. PMID- 6285239 TI - [Multiple sclerosis--an enigmatic disease]. PMID- 6285240 TI - [Hepatitis A antibodies in young women in Lower Saxony. Results of a serologic study on dried blood samples from the neonatal screening program for inborn errors of metabolism]. PMID- 6285241 TI - [Ifosfamide versus ifosfamide + CCNU in the treatment of inoperable small cell carcinoma of the lung. A clinical study]. AB - 55 selected patients with inoperable small cell carcinoma of the lung have been induced in a randomized prospective trial in order to compare the efficacy of ifosfamide monotherapy with the combination of ifosfamid + CCNU. Initially a control group, treated with symptomatic therapy only, was also included but preliminary analysis revealing that results were unfavourable in comparison to those of the chemotherapy group; this treatment arm was omitted. 53 of 55 patients were evaluable. In group 1 (symptomatic therapy) results were better as to patients' quality of life, but no tumor remissions have been observed and survival times were significantly shorter (15 to 122 days; geometric mean 42.35 days). In group 2 (ifosfamide monotherapy) one total remission (lasting 15 months) and 2 partial remissions (lasting from 3 weeks to 4 months) have been achieved; in this group survival was 3 to 473 days (geometric mean 110.15 days). In group 3 (ifosfamide + CCNU) 7 partial remissions have been observed (3 weeks to 4 months), survival was 17 to 477 days (geometric mean 107.32 days). The combination of ifosfamide with CCNU did not reveal any advantage in comparison to ifosfamide monotherapy. PMID- 6285242 TI - [Improved prognosis in Wilms' tumor due to adjuvant combination drug therapy]. AB - 78 children with Wilms' tumor stage I--IV diagnosed since 1960 are presented. There were two groups of patients: one group consisting of 35 patients which received adjuvant chemotherapy for 1 year, the other group consisting of 43 patients which received no chemotherapy. Surgical excision and irradiation was identical in both groups. The prognosis was greatly improved by adjuvant chemotherapy: 4-year survival rates increased from 25 to 69%. Late side effects from radiation and chemotherapy were noted: 15/78 patients suffered from scoliosis and all patients treated by chemotherapy had a decreased lymphotoxin activity over years. The following factors appear to be related to prognosis: the extent of disease in patients determined by better techniques, histopathology of the tumor, and the nature of treatment. PMID- 6285243 TI - [Radionuclide therapy of malignant and benign extra-thyroid diseases]. AB - Basis, indications and results of the endolymphatic therapy with radionuclides, of the selective therapy with radiophosphorus and radiostrontium and of the intraarticular and endocavitary therapy with radiocolloids are described. Future scientific engagement and intensive basic research in radionuclide therapy is required. However, because of the expected therapeutic profit the efforts seem to be justified. PMID- 6285244 TI - [Primary liver carcinoma. Results of 268 autopsies]. AB - 268 patients with primary liver cancer have been autopsied: 81% had hepatomas, 14% cholangiocellular carcinomas, 5% mixed carcinomas (hepatocholangiocellular) and 1% had mixed tumors (hepatoma and angiosarcoma). In contrast to cholangiomas, hepatomas were found more often in males than in females. The age peak of primary liver cancer was between 61 and 70 years. 79% of hepatomas developed in a cirrhotic liver, 71% of cholangiomas were without cirrhosis of the liver. Metastases were found in 66-67% of hepatomas and cholangiocellular carcinomas, especially in blood vessels, lymph nodes, lungs and within the liver. PMID- 6285245 TI - Isolation and characterization of the growth-stimulating factor in supernatants of herpes simplex virus type 2 infected cells. AB - Lytic infection of CV1 cells with herpes simplex virus type 2 does not stimulate ornithine decarboxylase activity and there is no correlation between polyamines and a growth-stimulating factor (GSF) which is present in the supernatant of the cultures. The factor was partially purified by gel filtration on Sephadex G-50 and polyacrylamide gel electrophoresis. Gel filtration as well as dialysis through membranes with different pore diameters indicated a molecular weight between 3,500 and 10,000 daltons. GSF is not extracted from aqueous solutions by diethyl ether. The activity is partially sensitive to trypsin digestion and is completely destroyed by periodate oxidation. These results suggest that GSF is a glycoprotein or a glycopeptide whose mitogenic activity resides mainly in the sugar moiety. PMID- 6285246 TI - Diamond knife. AB - We present a new diamond knife which allows for reasonably precise incisions in cornea or sclera. The knife may be ultrasonically cleaned and is sharper than any metal knife whose edge we have examined to date by scanning electron microscopy. The edge is approximately 0.1 micron in width, compared to 1-5 micron width edges of most metal knives. We feel that this prototype will allow investigators to recommend special modifications to the manufacturer of their own choice. PMID- 6285247 TI - Investigations of the enzymes involved in the fructose breakdown in the cattle lens. AB - By introducing fructose into the glycolysis, it is possible to stimulate ATP formation. As is the case in animal experiments, in human lenses, too, the first step in the phosphorylation to fructose-1-phosphate via the enzyme ketohexokinase. The present investigation deals with the question whether enzymes present in the lens are responsible for the further steps in fructose degradation. Particularly the aldolase isoenzyme C splits fructose-1-phosphate into glyceraldehyde and dihydroxyacetone phosphate in the same way as in glucose catabolism. Dihydroxyacetone phosphate can further be directly degraded and thus utilized to ATP formation. From glyceraldehyde, glycerol (aldose reductase) or glycerate (aldehyde dehydrogenase) can be formed. The presence of triosekinase, which phosphorylates glyceraldehyde directly to glyceraldehyde-3-phosphate, could only be determined in the lens tissue of young animals. The presence of glycerokinase (glycerol leads to glycerophosphate) could not be verified. Thus, in the lens tissue 1 ATP molecule net per fructose molecule can be formed. In older age, the glucose breakdown is limited by hexokinase and phosphofructokinase, so that the glucose, after transformation via the sorbitol pathway to fructose, can also be utilized for the energy metabolism. PMID- 6285248 TI - Carotid sheath tumor--a diagnostic challenge. AB - The tumor within the carotid sheath described in this article is unusual in that its clinical presentation mimicked a deep-lobe parotid tumor. Despite extensive investigation with conventional sialography, technetium pertechnetate scintigraphy, and computerized axial tomography, the correct preoperative diagnosis was not established. The value of CAT scanning in the diagnosis of parapharyngeal tumors is demonstrated. PMID- 6285249 TI - [Adenoid cystic carcinoma of the salivary glands in patients in the Department of Otolaryngology, Medical Academy in Warsaw (author's transl)]. PMID- 6285250 TI - [25-hydroxy-vitamin-D in serum of newborns and infants during continuous oral vitamin D treatment (author's transl)]. AB - Using the Haddad modified method, 25-OH-D were measured in the blood of the umbilical cord of 29 infants and in peripheral serum after 6 weeks. 16 infants were given a daily dosage of 1000 I. E., 13 infants 500 I. E. vitamin D against rickets. Further they were fed with an adapted milk containing 400 I. E. vitamin D/1. The mean cord serum values were 13 and 15 ng/ml. After treatment with 1000 I. E., 25-OH-D values around 54 ng/ml were measured after 6 weeks and under 500 I. E. daily, values of 37 ng/ml, respectively. Treatment using a dosage of 500 I. E. vitamin D combined with feeding with vitamin D fortified milk seems adequate, to prevent vitamin D depletion. PMID- 6285251 TI - [Transient vitamin-D dependent rickets? a case report differential diagnosis (author's transl)]. AB - The clinical course of a 9-month-old boy with Vit. D resistant rickets receiving treatment course 1-Alpha-hydroxycholecalciferol is described. Patient's response to therapy was excellent, after regression of bone changes and normalization of serum parameters drug treatment was discontinued for differential diagnostic considerations. Even several months later no relapse occurred. Due to this fact and the high concentration of serum 1,25-dihydroxycholecalciferol concentration during therapy the very rare pseudovitamin-D-deficiency type II, which is characterized by a bone receptor defect, was assumed. In contrast, successful treatment with low-dose 1 Alpha-OHCCF indicates that the underlying defect is pseudovitamin-D-deficiency rickets type I, however this form would require lifelong therapy. The differential diagnosis based on recent knowledge of Vitamin D-metabolism is discussed. PMID- 6285252 TI - A bioenergetic basis for peripheral nerve fiber dissociation. AB - The selective vulnerability of myelinated axons in lesions of peripheral nerve is incompletely understood and appears somewhat at variance with the energy conservation attached to saltation in these fibers. We evaluated the relative energy requirement of resting A and C fibers in rabbit vagus nerve by measuring the amplitude of the components of the compound action potential at 5-10 min intervals during incubation in Ringer-bicarbonate solutions containing 0-20 mM glucose. In nerves in which the perineurial sheath was retained intact the A components remained at control amplitude with 20 mM glucose but, after a plateau period, declined increasingly rapidly with 5, 2, 1, 0.5 and 0 mM glucose. 2mM glucose sufficed to maintain control amplitude of the C fiber component. In desheathed nerves the A component remained at control amplitude with 5 mM glucose but declined increasingly rapidly with 2, 1, 0.5 and 0 mM glucose; 0.5 mM glucose sufficed to maintain control amplitude to C fibers. The depressed potentials generally recovered incompletely after transfer to glucose 5 mM (desheathed) or 20 mM (sheathed); however, the partial recovery was more rapid and more nearly complete in the C fiber group than in the A fiber group (P less than 0.05). The data demonstrate that resting A fibers are much more susceptible to energy lack in vitro than resting C fibers. This suggests that deprivation of energy may be a factor in the preferential destruction of large fibers, termed fiber dissociation, which characterized several syndromes of chronic pain. PMID- 6285253 TI - [Perchlorosoluble glycoproteins and myocardial infarct: modifications of the carbohydrate moiety (author's transl)]. AB - Protein, hexose, amino sugar and sialic acid levels were determined in the perchlorosoluble fraction of serum from patients with myocardial infarct. Patients have significantly elevated mean values for all components. The hexose/perchlorosoluble protein and amino sugar/perchlorosoluble protein ratios were not significantly different when compared in normal and patient sera but the sialic acid/perchlorosoluble protein ratio was higher in patients with myocardial infarct. This change in sialic acid content could be related to an increased sialylation of glycoproteins normally present in serum. Particularly, an orosomucoid fraction with a higher content in sialic acid than that of the normal glycoprotein has been identified. PMID- 6285254 TI - Collagenolytic activity by malignant tumours. AB - Five carcinomas and 5 sarcomas were investigated in relation to their production of neutral proteases capable of digesting polymeric collagen. The carcinomas were far more active than the sarcomas but all the malignant tumours produced enzymes which were capable of causing collagenolysis in vitro. collagenolytic enzymes were recovered from extracts of neoplastic cells from long-term culture, from the media in which these cells were cultured, from the media of mixed cell cultures (neoplastic, stromal and inflammatory cells from minced tumours), and from normal fibroblasts cultures. In contrast to the cultures of non-neoplastic fibroblasts, the tumour cells produced active enzymes, since limited proteolysis with trypsin or treatment with p-aminophenyl-mercuric acetate (APMA) caused no increase in enzyme activity. These tumours possess collagenolytic ability in vitro which may be partly responsible for their invasive nature in vivo. PMID- 6285255 TI - Primary colo-rectal linitis plastica type of carcinoma: report of two cases and review of the literature. AB - Linitis Plastica type of carcinoma can arise primarily in the colon or rectum. Fifty-two cases have been reported in the English medical literature over a 50-yr period. Two new cases are described. One patient had primary linitis plastica of the ascending colon, the other being in the rectum. Both patients died of widespread malignancy within 10 mth of diagnosis, the fate of most of the reported cases. This rare manifestation of colo-rectal carcinoma carries an ominous prognosis. PMID- 6285256 TI - Autopsy findings in bone marrow transplantation. AB - Autopsies were performed on 2 patients with aplastic anaemia and 7 with acute leukaemia dying after bone marrow transplantation. Neutropenic enterocolitis was found in 2 of the 3 early deaths occurring before marrow engraftment and was related to radiation or cytotoxic drug damage to the bowel mucosa in the presence of profound neutropenia, allowing infection by bowel organisms. Cytomegaloviral infection was universal in engrafted patients. One had cytomegaloviral (CMV) pneumonia, one CMV hepatitis and enteritis and one CMV enteritis. Three patients had occasional CMV inclusions in various organs without obvious harmful effects. One nonengrafted patient also had CMV pneumonia. Graft versus host disease (GVHD) was a significant finding in 4 engrafted patients. This was difficult to separate histologically from the effects of CMV in the bowel, but easier in liver and skin. The skin changes of GVHD were the most easily interpretable. Interstitial pneumonia was due to CMV in one nonengrafted and one engrafted patients and had no obvious infective cause in 2 engrafted patients. The presence of bizarre epithelial cells in the lungs of these patients suggested an aetiological role for radiation or cytotoxic drugs. Modification of the conditioning regimen may reduce tissue damage and lessen many of these side-effects. PMID- 6285257 TI - Wilms' tumor metastatic to the mediastinum. AB - We report two children with mediastinal metastasis of Wilm's tumor. One case presented with a right hilar mass and the other with a posterior mediastina mass. In each instance, no additional radiologic evidence of metastasis was found. Metastasis of Wilms' tumor to the mediastinum is uncommon, usually occurring with radiologic evidence of either additional metastases or direct cephalad growth of the primary tumor. PMID- 6285258 TI - CT localization of occult secretory tumors in children. AB - Three seemingly occult secretory tumors in children (pheochromocytoma, ganglioneuroblastoma and islet cell carcinoma) were localized within the abdomen by computed tomography after other diagnostic imaging procedures had failed. The superb density resolution and tomographic formating of CT images make CT uniquely suited for the demonstration of small abdominal lesions. CT is recommended as a primary imaging modality for secretory tumors in children. PMID- 6285259 TI - Echographic aspects of hepatic metastases of nephroblastomas. AB - We report a retrospective study of 293 cases of nephroblastoma treated at the Gustave Roussy Institute (I.G.R.) from 1971 to 1980, and 39 similar cases treated at the Curie Institute (I.C), from June 1977 to June 1981. Twenty-two cases of hepatic metastases were found. We were primarily interested in the echographic aspects of the lesions and particularly in their evolution. Despite the fact that the patients came from different centers with varying types of work-up, ten cases were fully documented and are presented in detail. They show the use to which ultrasound may be put. PMID- 6285260 TI - Renal oxygen consumption and sodium reabsorption during isotonic volume expansion in the developing rat. PMID- 6285261 TI - Superoxide generation by human fetal granulocytes. AB - Phagocytic leukocytes such as granulocytes and macrophages form an essential part of human host defense against infection by invading microorganisms. The present study demonstrates that normal fetal granulocytes undergo normal activation (i.e., the transition from the resting state to the respiratory burst) with a 75 +/- 23 sec lag time and generate superoxide at 6.7 +/- 2.3 nmole 02-/min/10(6) cells, a rate similar to that of adult granulocytes. Fetal granulocytes also resemble adult cells in the response of the superoxide generating system to temperature change and to the chemical inhibitors N-ethyl maleimide and 2 deoxyglucose. They show the same pattern of differential effects of these perturbations on the lag time and on the rate of superoxide generation as that previously described for adult granulocytes. The process of activation, expressed as the lag time, shows no inhibition at a high temperature that diminishes by 64% the rate of superoxide production. The metabolic inhibitor 2-deoxyglucose affects only the rate and not the lag time. Both cyanide (in the presence of 2 deoxyglucose) and the sulfhydryl reagent N-ethyl maleimide inhibit the rate equally when added before or after completion of the activation process. PMID- 6285262 TI - Ontogeny of beta-adrenergic receptors in the rat lung: effects of hypothyroidism. AB - Beta-Adrenergic receptors were identified in membrane fractions of rat lung with the beta-adrenergic antagonists (-)-[3H]-dihydroalprenolol (-)-[3H]DHA) and (+/-) [125I]-iodohydroxybenzylpindolol ((+/-)-[125I]HYP). Binding capacity (Bmax) for ( )-[3H]DHA increased progressively from 46 +/- 7 on day 18 of gestation to 510 +/- 70 femtomoles . mg-1 protein (mean +/- S.D.) on postnatal day 28, at which time adult Bmax was attained. An increase in (-)-[3H]DHA binding capacity of the lung was observed between postnatal days 15 and 28, during the known period of increased thyroid gland secretory activity, serum triiodothyronine (T3), and thyroxine (T4) concentrations in the rat. We therefore studied lung beta adrenergic receptors in rat pups made hypothyroid with propylthiouracil (PTU) (in utero and postnatally) compared to normal age-matched control pups and to euthyroid pups which were treated with PTU but were also injected daily with thyroxine (T4-treated). Hypothyroid pups grew nearly normally until postnatal day 15 but grew poorly thereafter; but day 28 somatic and lung weight, lung DNA, and protein were markedly decreased in hypothyroid pups as compared to controls. Pulmonary beta-adrenergic receptors were similar in hypothyroid pups and controls on day 15, but were markedly decreased in hypothyroid pups on day 28 (294 +/- 57 versus 489 +/- 82 femtomoles . mg-1 protein in T4 treated euthyroid controls). Treatment of the hypothyroid pups with T4 on day 25 significantly increased lung beta-adrenergic receptors to near normal concentrations by day 28. We conclude that thyroid hormones or thyroid dependent factors enhance pulmonary beta adrenergic receptor synthesis and that thyroid hormone is required for the normal postnatal maturation of the beta-adrenergic receptor system in the rat lung. PMID- 6285263 TI - Brain and visceral involvement during congenital cytomegalovirus infection of guinea pigs. AB - The virologic and histologic characteristics of congenital cytomegalovirus infection (CMV) were defined in 65 neonatal guinea pigs born from 27 mothers infected pregnancy. Infectious virus or tissue lesions were present in 54% of the neonates tested. Guinea pig CMV was detected most often in the salivary glands (72%) and spleen (33%) of infected guinea pigs. Less frequently, virus was also detected in the brain, lung, pancreas and liver. Tissue lesions were most frequently observed in the brain and kidney, but also occurred in the salivary glands, liver, pancreas, thymus and spleen. The histopathology was identical to that observed in infants with congenital CMV infection. Infectious virus and histopathology were present in newborn guinea pigs born from mothers infected at any time during gestation. Newborns from mothers infected during early stages of gestation and virus present most frequently in the salivary glands, whereas offspring of mothers infected in late pregnancy had virus present in several tissues. Acute maternal guinea pig CMV infection produced generalized CMV infection of the offspring which was followed by persistent infection in neonatal salivary glands. Lesions remained present in several neonatal tissues including the brain. The long term consequences of such lesions in affected guinea pigs remain to be determined. The results of the study emphasize the similarities between human congenital CMV infection and congenital guinea pig CMV infection, thereby underlining the utility of this animal model as a means of understanding human congenital CMV infection. PMID- 6285265 TI - [Electron microscopy in the diagnosis of congenital cytomegaly]. PMID- 6285266 TI - [Diagnostic efficacy of the determination of antibodies against herpes simplex virus type I in the cerebrospinal fluid and blood of children with meningoencephalitis]. PMID- 6285264 TI - beta-Adrenergic receptors and catecholamine sensitive adenylate cyclase in developing rat ventricular myocardium: effect of thyroid status. AB - The present study was designed to describe the relationships among thyroid status, myocardial growth and myocardial beta-adrenergic receptors in the developing rat ventricle. In normal rat myocardium the beta-adrenergic binding capacity (Bmax) for (-)-[3H] DHA decreased with increasing age and heart size. In order to determine the effect of thyroid status on ventricular growth characteristics and beta-adrenergic receptors, animals were rendered: (1) hypothyroid with propylthiouracil (PTU), (2) euthyroid with PTU and daily thyroxine (T4) replacement, (3) hyperthyroid for several days with daily thyroxine injections or (4) normal controls with sham saline injections. Growth characteristics were similar in euthyroid and normal rat myocardium; ventricular weight, protein and DNA content were similar at postnatal days 5, 15 and 28. Growth in hypothyroid pups was normal until postnatal day 14 at which time the heart weight and protein content were significantly lower than in normal or euthyroid pups, whereas the number of beta-adrenergic receptors was decreased in hypothyroid myocardium at all ages studied. On postnatal day 5 the (-)-[3H]DHA binding (Bmax) was 37 +/- 9 in hypothyroid myocardium compared to 63 +/- 8 fmole per mg protein mean +/- S.D. in euthyroid myocardium. The function of the beta adrenergic receptors was also decreased in hypothyroid as compared to euthyroid or normal myocardium as demonstrated by a decrease in maximal catecholamine sensitive adenylate cyclase activity in myocardial membranes at 28 days of age. Treatment of hypothyroid or normal pups with T4 resulted in an increase in heart size, protein content and beta-adrenergic receptors. Ventricular DNA content, which describes hyperplastic growth, was not decreased in hypothyroid rats demonstrating that postnatal hypertrophic but not hyperplastic ventricular growth is dependent on thyroid hormone. PMID- 6285267 TI - Parotid acinar cells: ionic dependence of isoprenaline-evoked membrane potential changes. AB - The effect of microionophoretic application of isoprenaline on membrane potential and resistance of mouse parotid acinar cells was investigated. For measurements of membrane resistance and the isoprenaline equilibrium potential (Eiso), two microelectrodes were inserted into neighbouring communicating cells. Passing direct current through one of these electrodes, the resting potential could be set at desired levels and Eiso was determined by plotting the relation between the size of the isoprenaline-evoked potential change and the resting potential. Simple depolarizations were found at relatively high resting potentials, while biphasic potential changes in response to isoprenaline (hyperpolarization followed by depolarization) were observed at low resting potentials. Both depolarizing and hyperpolarizing responses to isoprenaline were accompanied by a reduction of membrane resistance. The isoprenaline equilibrium potential in the initial phase of the response was about -53 mV, but had a value of about -24mV in the delayed phase. The initial isoprenaline-evoked potential change was sensitive to alterations in extracellular Na, K and Cl concentrations. The delayed depolarizing response to isoprenaline was markedly reduced by replacing extracellular Na by Tris or extracellular Cl by SO4. These results indicate that isoprenaline opens up conductance pathways permeable to Na and K. PMID- 6285268 TI - The effect of colchicine on neuromuscular transmission in the frog during repetitive stimulation. AB - When colchicine 10(-4) mol . l-1 was applied at the beginning of repetitive stimulation of the nerve at 10 s-1, the facilitation was markedly inhibited. The quantal content showed a very slight increase and its maximum value was reached later. The maximum frequency of spontaneous release was also reached later in the presence of colchicine than in the absence of the drug. In addition, the synaptic delay was much more pronounced in the presence of colchicine than in the control experiment. The results suggest that a partial block by colchicine of the release process in the nerve terminal occurs. This effect may be due to the action of the drug on the nerve terminal membrane. The results cannot exclude the possibility that colchicine interferes with the transport of vesicles towards the sites of release located on the membrane. PMID- 6285269 TI - Properties of a Ca2+ activated K+ conductance in Helix neurones investigated by intracellular Ca2+ ionophoresis. AB - 1. Membrane current responses produced by intracellular ionophoretic injection of Ca2+ ions into voltage clamped Helix neurones were investigated. 2. The Ca2+ activated current depends on the intensity and duration of the Ca2+ injection as well as on the membrane potential. The current-voltage relation is non-linear and shows strong outward rectification. 3. The Ca2+ activated current reverses near the equilibrium potential for K+ ions and the associated current fluctuations disappear at the K+ reversal potential, which indicates a single current source, probably activation of K+ channels. 4. The Ca2+ activated K+ conductance is voltage dependent and changes e-fold per 31 mV membrane depolarization. 5. Extracellular tetraethylammonium, gallamine and quinidine block the Ca2+ activated K4 current. Extracellular 4-aminopyridine has no blocking effect. 6. Sustained injection of high amounts of Ca2+ irreversibly suppresses the outward current. 7. The K+ outward current evoked by repeated Ca2+ injection exhibits summation and facilitation. PMID- 6285270 TI - Noise and relaxation measurements of the Ca2+ activated K+ current in Helix neurones. PMID- 6285271 TI - Effect of kidney denervation on cortical iso-converting enzyme activity in the rat. PMID- 6285272 TI - [Angiographic findings of small hepatocellular carcinoma (author's transl)]. PMID- 6285273 TI - [Studies of absorbed dose determinations and spatial dose distributions for high energy proton beams (author's transl)]. PMID- 6285274 TI - [Evaluation of arterial embolization therapy for hepatocellular carcinoma by ultrasonography (author's transl)]. PMID- 6285275 TI - [Peripheral neuropathies and isaxonine]. PMID- 6285276 TI - [Peripheral neuropathies. Current data concerning nerve regeneration (author's transl)]. AB - The peripheral neuropathies in humans could be classified both with clinical criteria and pathological aspects. Between these, only axonal neuropathies are subject to regeneration. Axonal neuropathies present two aspects: wallerian degeneration and dying-back neuropathy. Dying-back neuropathy is the commonest type of human neuropathy but wallerian degeneration has been the most studied model of axonal neuropathy. The regeneration in this case lead to two questions: what is the signal for regeneration and what are the factors which improve the nerve growth? PMID- 6285277 TI - [Pharmacological basis of the action of isaxonine (author's transl)]. AB - The author reviews previous therapeutic attempts to stimulate regeneration of peripheral nerves. Since 1971 he has been studying a synthetic compound, isaxonine, which in rats accelerates nerve regeneration and functional recovery. Stimulation of axonal regeneration and collateral sprouting has been demonstrated in tissue cultures. The therapeutic activity of the compound has been demonstrated in man in leprous neuropathy and in vincristine neuropathy. Isaxonine has specific affinity for peripheral nerves. Its acts directly on the neuron or indirectly by stimulating production of a growth factor remains unknown. In vitro and in vivo studies isaxonine have shown that isaxonine antagonizes the noxious effects of Vinca alkaloids on neurotubular structure. Research in progress suggest a possible action of this new compound in other diseases related to tubulin anomaly. PMID- 6285278 TI - [Effects of isaxonine on experimentally induced vincristine neuropathy in rabbits (author's transl)]. AB - The effect of isaxonine on neuropathy induced by vincristine in the rabbit was studied using clinical, electrophysiological and histological parameters. Isaxonine protects against early adverse reactions of vincristine administration such as decrease in food consumption and body weight, an alopecia. Motor disturbance was reduced. This protective effect is confirmed by electrophysiological tests (motor latency, muscular strength of tibialis anterior muscle). Isaxonine decrease the incidence of the neuropathy induced by vincristine in the rabbit. PMID- 6285279 TI - [Protective effect of isaxonine against vincristine-induced neuropathy (author's transl)]. AB - A double-blind survey versus a placebo was carried out with C.P.V. protocole on 25 patients treated for lymphosarcoma or reticulosarcoma; its aim was to appraise the preventive action of isaxonine on the appearance of neuropathy induced by vincristine. The frequency of such neuropathy was clearly lower in the isaxonine group (3/10) than in the placebo group (8/10) in a statistically significant manner. The protective effect of isaxonine was particularly striking on tendon reflexes and on the number of motor units recorded at musculus extensor digitirum pedis brevis; on the contrary, the patients treated with the placebo showed real denervation. PMID- 6285280 TI - [Controlled trial of isaxonine in traumatic or ischaemic (compression) neuropathies (author's transl)]. AB - Fifty patients showing one or more impairments to large nerve trunks due to trauma or ischaemia received, after drawing lots, either isaxonine or a placebo. The evaluation criteria of the recovery were both clinical (testing of muscles, sensory disorders, vegetative disorders, tendon reflexes, tonicity, amyotrophy) and electrophysiological (electromyogram, conduction velocity, distal latency time, H reflex and H/M index). This study shows two major elements: the action of isaxonine shortens healing time (45-90 days instead of 120-150 days) and isaxonine allows neurological recovery to resume at neurologic level whereas the situation was dating back more than three months. PMID- 6285282 TI - [Anaplastic small cell bronchial cancer]. PMID- 6285281 TI - [Acceleration by isaxonine of muscle reinnervation. Electrophysiological demonstration in leprotic neuropathy (author's transl)]. AB - On a model of compressive human neuropathy--leprous neuropathy occuring below an osteoligamentary canal following a process of sclerosis--a double-blind survey of isaxonine versus a placebo was undertaken after matching of patients to obtain similar pairs. After four months of treatment with isaxonine, the electrophysiological tests (EMG of the anterior tibialis and of the abductor digiti minimi muscle) showed a very significant improvement in the graphs of patients receiving the active agent, as compared to a lack of improvement, even an aggravation for those patients receiving the placebo. It may thus be concluded that muscular reinnervation has occurred, either by sprouting of the remaining healthy fibers or by regeneration of the damaged axons, or by both phenomena simultaneously. PMID- 6285283 TI - [Acute cytomegalovirus pancreatitis with a favorable outcome in a renal transplant patient]. PMID- 6285284 TI - [Immune deficiency and Kaposi's disease in 2 young male homosexuals]. PMID- 6285285 TI - The anticodon of the maize chloroplast gene for tRNA Leu UAA is split by a large intron. AB - The maize chloroplast gene encoding tRNA Leu UAA has been sequenced. It contains a 458 base pair intron between the first and second bases of the anticodon. The tRNA is 88 nucleotides long (the 3'-terminal CCA sequence included which, however, is not encoded by the gene) and differs in only four nucleotides (modified nucleotides are not considered) from the corresponding isoacceptor from bean chloroplasts. The unusual position of the intron in this maize chloroplast tRNA gene suggests a splicing model different from that generally accepted for eukaryotic split tRNA genes. PMID- 6285286 TI - Non-random localization of ribonucleoprotein (RNP) structures within an adenovirus mRNA precursor. AB - Heterogeneous nuclear protein complexes (hnRNP) containing the precursor RNA from the adenovirus early region 2 were analysed to determine the specificity of protein-RNA interaction. RNA precursor sequences were present in isolated hnRNP complexes and endogenous 30S particles. At least 20-40 bases long fragments were protected when RNase A was used to remove unprotected RNA sequences in hnRNA complexes. Similarly around 40 bases of RNA were protected in 30S particles. These sequences represent discrete regions of the adenovirus genome. Especially sequences complementary to the EcoRI-F fragment encoding the first leader and the major intron for the DNA binding protein (DBP) RNA precursor, were analysed in detail. Tentatively, sequences resistant to RNase A were located in the middle of the intron and at the splice-donor junction of the first leader of the DBP precursor RNA. The same sequences were identified irrespective whether hnRNP complexes or 30S particles were used suggesting that 30S particles originate from hnRNP complexes. A 38.000 dalton protein appears to be in direct contact with RNA sequences complementary to the EcoRI-F fragment. PMID- 6285287 TI - DNAase I sensitivity of genes expressed during myogenesis. AB - Cultures of a rat myogenic cell line were used to examine the question of whether in proliferating precursor cells genes which are programmed to be expressed later in development, in the same cell lineage, differ in DNAase I sensitivity from genes which are never expressed in these cells. Nuclei isolated from proliferating mononucleated myoblasts, differentiated cultures containing multinucleaged fibers, and rat brain, were treated with DNAase I. The sensitivity of the genes coding for the muscle-specific alpha-actin, myosin light chain 2 and the nonmuscle beta-actin was measured by blot hybridization of nuclear DNA with the corresponding cloned cDNA and genomic DNA probes. The sensitivity of these genes was compared to that of a gene not expressed in the muscle tissue. The results showed that in the muscle precursor cells, the potentiality of tissue specific genes to be expressed is not reflected in DNAase I sensitivity. The changes which render these genes preferentially sensitive to DNAase I take place during the transition to terminal differentiation. The results showed also that the region of DNAase I sensitivity of the alpha-actin gene in the differentiated cells ends between 40 to 700 bp 5' to the structural gene. No DNAase I hypersensitive site was detected 5' to the alpha-actin gene. PMID- 6285288 TI - Molecular cloning and analysis of yeast gene for cycloheximide resistance and ribosomal protein L29. AB - A cosmid clone bank of yeast DNA has been used to isolate the cycloheximide resistance gene cyh2 of Saccharomyces cerevisiae. A cosmid carrying this gene was identified by cross hybridization to another cloned gene, tsm437. The two genes, which are tightly linked genetically are both present on a 31 kb segment of cloned DNA. The cyh2 gene encodes ribosomal protein L29, a component of the large subunit. Blot hybridization analysis reveals that this gene is present as a single copy in the yeast genome, unlike many other yeast ribosomal protein genes which appear to be duplicated. The cyh2 gene also appears to contain an intervening sequence, a characteristic common to most yeast ribosomal protein genes that have been cloned. PMID- 6285289 TI - Structure and expression of a Trypanosoma brucei gambiense variant specific antigen gene. AB - The expression-linked copy of the T. b. gambiense variant specific antigen gene LiTat 1.6 is transposed in a 20 kb DNA region devoid of restriction sites, located near a chromosome end. This expression site is very similar to that of T. b. brucei variants 117, 118 (1) and AnTat 1.8. In the basic copy, the transposable element (TE) is flanked by repetitive sequences; it includes the gene copy as well as a sequence of 0.9 to 2.1 kb (probably around 1.1 kb) long, upstream from the gene. Probes derived from the 5' part of the TE specifically reveal three polyadenylated transcripts of 4.2, 1.45 and 0.85 kb, respectively, distinct from the 2.1 kb mRNA. The amount of the 4.2 kb sequence is probably less than 0.01% of total trypanosome RNA. Whereas the mRNAs coding for the three isotypic antigens AnTat 1.8 (T. b. brucei), 12.2 (T. b. rhodesiense) and 3.3 (T. evansi) are recognized by LiTat 1.6 probes extending into the 3' half of the transposed sequence, the 5' genomic probes do not hybridize with any of these RNAs. These observations suggest that the LiTat 1.6 gene could be first transcribed in a large precursor molecule. This precursor would be rapidly processed, loosing a large portion of less conserved sequence from its 5' half. Our data are compatible with a model in which the promoter would be provided by the expression site. PMID- 6285290 TI - KpnI families of long, interspersed repetitive DNAs in human and other primate genomes. AB - KpnI restriction of DNAs from all anthropoid primates studied releases a conspicuous series of segments representing families of long, interspersed repetitive DNAs termed here the KpnI 1.2, 1.5, 1.8 and 1.9 kb families. Human KpnI 1.2 to 1.9 kb segments representative of these families were isolated and separately cloned in the KpnI site of a plasmid pBK5, specially constructed for this purpose. The KpnI clones did not cross-hybridize with cloned, primate alphoid sequences, suggesting that the KpnI families represent sequences separate and distinct from the alphoid DNAs. Secondary restriction analyses of cloned KpnI segments demonstrated microheterogeneity among individual members within the same KpnI family. Autoradiograms of capuchin monkey, AGM and human DNA cleaved with HaeIII, AluI or RsaI and hybridized to various cloned human KpnI sequences demonstrated a remarkable conservatism and relative simplicity in the organization of the KpnI families in the genomes of these widely divergent primates. The KpnI 1.2 kb and 1.5 kb families occur in high frequency (15%) among all plaques in two recombinant human genome libraries. Evidence is presented suggesting that the bulk of the KpnI families occur in the genome as clusters or congeries of higher molecular weight segments (greater than 2 kb) containing sequences homologous to the low molecular weight segments (1.2 to 1.9 kb). PMID- 6285291 TI - In vitro transcription by purified yeast RNA polymerase II. Coarse promoter mapping on homologous cloned genes. AB - Clones of the yeast Tyl element and 2 microns plasmid have been selectively transcribed in vitro by partially or completely purified yeast RNA polymerase II. Electrophoretic analysis of whole and restricted ternary transcription complexes allows the localization of the in vitro actively transcribed regions of the analyzed genes. The DNA regions that actively promote in vitro transcription correspond to the nucleotide sequence that in the Tyl element encompasses the in vivo transcription initiation sites and that in the 2 micrometer plasmid encompasses the starting codons of two oppositely oriented potential protein coding frames. The transcription assay that we describe herein may be applied to analyze rapidly the in vitro transcription of clones genes, to localize transcription initiation sites on supercoiled templates and to evaluate the differential in vitro promoter strength in RNA polymerase II served genes. Data obtained with RNA polymerase II at two different stages of purification are presented in parallel. Studies with a completely purified enzyme should certainly be preferred although the use of a partially purified RNA polymerase II may be convenient and may reveal factors which affect specificity. PMID- 6285292 TI - Nucleotide sequence definition of a major human repeated DNA, the Hind III 1.9 kb family. AB - A human Hind III 1.9 kb repeated DNA fragment was isolated and cloned in pBR322. A cloned member that hybridized predominantly to the 1.9 kb Hind III band in a digest of whole human DNA was chosen for sequencing. It is an 1894 bp fragment that shows no significant internal repeats. Few pCG residues are observed in the sequence and there are numerous stop codons. Detailed sequence comparisons confirm this is a novel class of repeats that is not related to previously characterized human satellite DNAs or Alu sequences. At least a portion of the sequence described is conserved in evolution. PMID- 6285293 TI - Genomic representation of the Hind II 1.9 kb repeated DNA. AB - The genomic representation and organization of sequences homologous to a cloned Hind III 1.9 kb repeated DNA fragment were studied. Approximately 80% of homologous repeated DNA was contained in a genomic Hind III cleavage band of 1.9 kb. Double digestion studies indicated that the genomic family, in the majority, followed the arrangement of the sequenced clone, with minor restriction cleavage variations compatible with a few base changes. Common restriction sites external to the 1.9 kb sequence were mapped, and hybridization of segments of the cloned sequence indicated the 1.9 kb DNA was itself not tandemly repeated. Kpn I bands which were homologous to the sequence contained specific regions of the repeat, and the molecular weight of these larger fragments could be simply explained. Mapping of common external restriction sites indicated that in some but not all cases the repeat could be organized in larger defined blocks of greater than or equal to 5.5 kb. In some instances, flanking regions adjacent to the repeat may contain common DNA elements such as other repeated DNA sequences, or possibly rearranged segments of the 1.9 kb sequence. It is suggested that although the 1.9 kb sequence is not strictly contiguous, at least some of these repeated sequences in the human genome are arranged in clustered or intercalary arrays. A region of the 1.9 kb sequence hybridized to a mouse repeated DNA, indicating homology beyond the primates. PMID- 6285294 TI - Nucleotide sequence of the rrnG ribosomal RNA promoter region of Escherichia coli. AB - The primary structure of the promoter region for a ribosomal RNA transcription unit (rrnG) of Escherichia coli K12 has been determined. The sequence was obtained from 1 1.5 kbp EcoRI fragment derived from the hybrid plasmid pLC23-30. This fragment contains 455 bp preceding P1 of the rrnG promoter region and 674 bp of the rrnG 16S RNA gene. The sequence before the rrnG promoter region contains an open reading frame (ORF-BG) followed by a possible hairpin structure that resembles other known transcription terminators. The sequence of the rrnG promoter region is similar but not identical to that of rrnA and rrnB. Several minor differences between the sequences of the 16S RNA genes of rrnG and rrnB were also noted. In addition, sequences were found that could generate special structures involving the promoter regions of rrn loci. Such structures are described and their possible involvement in the regulation of ribosomal RNA synthesis is discussed. PMID- 6285295 TI - Nucleotide sequences of the murine retrovirus Friend SFFVp long terminal repeats: identification of a structure with extensive dyad symmetry 5' to the TATA box. AB - The two long terminal repeats (LTRs) of the integrated provirus of the polycythemia strain of the Friend spleen focus forming virus have been sequenced. Each of the identical LTRs is 514 nucleotides long and together confer on the provirus the features of a transposable element. They are terminated by perfect 11 base-pair inverted repeats, and the entire provirus is flanked by an apparent duplication of host DNA four nucleotides long. The assumed transcription regulatory sequences (Hogness-Goldberg box, CAAT box, polyadenylation signal) can be identified within the LTRs, as well as a region of an imperfect inverted repeat which extends from approximately 140 to 270 nucleotides 5' from the point of transcription initiation. This sequence may form a hairpin-type structure which might have some function in the promotion of transcription. PMID- 6285296 TI - Organization of the mitochondrial ribosomal RNA genes of maize. AB - The organisation of the mitochondrial ribosomal RNA genes in maize is described. Each of the rRNAs is encoded by a single gene. The 5S and 18S rRNA genes are close together, and separated from the 26S rRNA gene by 16 kb of DNA. There is no evidence of heterogeneity in this gene arrangement. PMID- 6285297 TI - Simple DNA sequences in homologous flanking regions near immunoglobulin VH genes: a role in gene interaction? AB - Five closely related immunoglobulin VH genes (subgroup II) were compared by sequencing of several kb of DNA. In three of the genes homology greater than 75% was found along an area of 4 kb that includes the coding region. The homology in flanking regions is only slightly lower than that in the coding sequences. Two other genes, which are located on the same EcoRI fragment, show high homology to the first three genes in the coding and immediately flanking regions. In more distant flanking regions no homology is found with the first three genes. This indicates that their evolutionary history differs from that of the other three genes. A region of simple DNA sequence composed of repetitive TCC and TCA elements was found at a distance of approximately 380 bp upstream from the initiator ATG of these VH genes. This region is the site where the two sets of genes abruptly start to diverge. The structure of the simple DNA sequence in the various VH genes suggests that it may be involved in gene interaction. We propose that both simple DNA sequences and homology in flanking regions serve a function in the correction of VH genes, which seem to be rather free to diverge and drift into pseudogenes. A correction mechanism may help this gene family to maintain its two major features, multiplicity and diversity. PMID- 6285298 TI - Multiple components in restriction enzyme digests of mammalian (insectivore), avian and reptilian genomic DNA hybridize with murine immunoglobulin VH probes. AB - High molecular weight genomic DNAs isolated from an insectivore, Tupaia, and a representative reptilian, Caiman, and avian, Gallus, were digested with restriction endonucleases transferred to nitrocellulose and hybridized with nick translated probes of murine VH genes. The derivations of the probes designated S107V (1) and mu 107V (2,3) have been described previously. Under conditions of reduced stringency, multiple hybridizing components were observed with Tupaia and Caiman; only mu mu 107V exhibited significant hybridization with the separated fragments of Gallus DNA. The nick-translated S107V probe was digested with Fnu4H1 and subinserts corresponding to the 5' and 3' regions both detected multiple hybridizing components in Tupaia and Caiman DNA. A 5' probe lacking the leader sequence identified the same components as the intact 5' probe, suggesting that VH coding regions distant as the reptilians may possess multiple genetic components which exhibit significant homology with murine immunoglobulin in VH regions. PMID- 6285299 TI - Accurate transcription of cloned Xenopus rRNA genes by RNA polymerase I: demonstration by S1 nuclease mapping. AB - We have demonstrated faithful transcriptional initiation of cloned Xenopus rRNA genes upon injection into Xenopus oocytes. This observation has been made possible by the use of an S1 nuclease assay that is both sensitive and quantitative. In order to detect rRNA synthesis from the injected template above the large background of rRNA endogenously present in oocytes, the divergence of ribosomal DNA sequences between two Xenopus species was utilized. Cloned X. laevis ribosomal DNA was injected into the nuclei of X. borealis oocytes. Total oocyte RNA was then isolated and hybridized to a radioactive DNA probe that over laps the 5' end of X. laevis rRNA; endogenous rRNA of the X. borealis oocytes does not hybridize to the probe. RNA/DNA hybrids were treated with S1 nuclease and protected fragments were sized by polyacrylamide gel electrophoresis. RNA made from the injected rDNA protects the same region of probe as does authentic X. laevis precursor rRNA. Thus, transcription appears to initiate on the cloned, microinjected X. laevis rDNA at the same site as is used in vivo. This synthesis is not impaired by coinjection of an amount of alpha-amanitin sufficient to inhibit RNA polymerase II and III; therefore the reaction is mediated by RNA polymerase I. The amount of transcription may be reproducibly quantitated and we have varied a number of parameters in order to maximize transcriptional expression of the injected rDNA. Eight independently isolated X. laevis rDNA clones as well as several subcloned initiation regions of these genes are all accurately transcribed at approximately equal efficiency. This assay should facilitate analysis of several aspects of rRNA transcription, including deleniation of the Xenopus RNA polymerase I promoter location. PMID- 6285300 TI - In vitro transcription of two Epstein-Barr virus specified small RNA molecules. AB - Cloned DNA from the EcoRI J fragment of EBV has been used as template for in vitro transcription experiments using cell-free extracts prepared from HeLa or KB cells. Two EBV specific RNAs each about 175 bases in length were synthesised and nuclease 51 mapping experiments determined that these in vitro products corresponded precisely to the in vivo species obtained from Raji cells. These two RNA molecules are transcribed by RNA polymerase III and in common with other pol III-synthesised RNAs the coding sequences contain intragenic control regions. The relative abundance of the two RNAs synthesised in vitro differs from that observed in vivo. PMID- 6285301 TI - Relative levels of methylation in human growth hormone and chorionic somatomammotropin genes in expressing and non-expressing tissues. AB - It has been shown that the extent of methylation of cytosine in vertebrate DNA is inversely correlated with gene expression. We studied cytosine methylation in and around the homologous human growth hormone (GH) and chorionic somatomammotropin (CS) genes to determine if these genes are undermethylated in DNA from tissues in which they are expressed (pituitary and placenta, respectively) compared to other tissues. Hpa II and Hha I (which cleave only unmethylated 5' CCGG 3' and 5' GCGC 3' respectively) and Msp I (which cleaves CCGG and CmeCGG) were used to digest DNA samples followed by gel electrophoresis, Southern transfer and hybridization with a GH cDNA probe. The extent of methylation of Hpa II and Hha I sites in the GH and CS genes was leukocyte much greater than pituitary greater than placenta = hydatidiform mole. Taken as a whole, our data support the hypothesis that undermethylation is a necessary but not sufficient condition for gene expression since placental and pituitary DNAs are less methylated than leukocyte DNA in this region. However, the correlation between gene expression and undermethylation is imperfect since (1) hydatiform mole DNA has a very similar methylation pattern compared to placental DNA even though moles make little or no CS and (2) the level of methylation of the GH gene compared to the CS gene does not vary in a tissue-specific manner. PMID- 6285302 TI - Effect of in vitro methylation at CpG sites on gene expression in a genome functioning autonomously in a vertebrate host. AB - The effect of in vitro methylation at the HpaII sites in polyoma DNA on viral gene expression and the maintenance of the methyl groups upon replication in vivo were examined. Most of the methylatable sites are located in the early region coding for the viral large T antigen which is essential for the replication and infectivity of the viral DNA. Methylated or mock-methylated polyoma DNA produced the same number of virus plaques appearing at the same time post-transfection in either case. The lack of effect on the infectivity of the viral DNA indicates that the expression of the T antigen gene was not inhibited by methylation. Replication in vivo of the DNA also resulted in a total loss of the methyl groups introduced in vitro. These results underscore basic differences between the behavior of an autonomously functioning papovavirus DNA and the animal cell DNA vis-a-vis methylation at CpG sites. These differences might be due to subtle variations in the mechanism of regulation of gene expression and replication in the two systems. PMID- 6285303 TI - The nucleotide sequences of two leghemoglobin genes from soybean. AB - We present the complete nucleotide sequences of two leghemoglobin genes isolated from soybean DNA. Both genes contain three intervening sequences in identical positions. Comparison of the coding sequences with known amino-acid sequences of soybean leghemoglobins suggest that the two genes correspond to leghemoglobin C2 and leghemoglobin C3, respectively. PMID- 6285304 TI - Chromatin diminution in Ascaris suum: nucleotide sequence of the eliminated satellite DNA. AB - We have determined the prototype sequence of the DNA which is eliminated in the course of chromatin diminution in Ascaris suum. This DNA which is virtually absent from somatic cells but retained in the germ line consists predominantly of highly repetitive sequences which are variants of an AT rich 123 base pair repeat unit. Both major and minor variants have been sequenced. The overall structure of this germ line limited DNA corresponds to the segmental organization characteristic of satellite DNAs. Possible correlations between the mechanism of chromatin diminution and some properties of the satellite sequence are discussed. PMID- 6285305 TI - Comparison of the nucleotide sequence of cloned human and guinea-pig pre-alpha lactalbumin cDNA with that of chick pre-lysozyme cDNA suggests evolution from a common ancestral gene. AB - Nucleotide sequence analyses of essentially full-length copies of human and guinea-pig pre-alpha-lactalbumin cDNAs contained within recombinant plasmids, (i) confirm the presence of 19 amino acid hydrophobic amino terminal peptide extensions encoded within each mRNA; and (ii) provides evidence for the existence of a minor variant of guinea-pig alpha-lactalbumin mRNA encoding a protein with a 36 residue carboxyl-terminal extension. Comparison of the nucleotide sequence within the coding region of the human, and the predominant guinea-pig pre-alpha lactalbumin mRNAs, with the analogous region of hen pre-lysozyme mRNA provides compelling evidence that all have evolved from a common ancestral gene. PMID- 6285307 TI - Primary and secondary structure specificity of the cleavage of 'single-stranded' DNA by endonuclease Hinf I. AB - The interaction of endonuclease Hinf I with single-stranded fd DNA was examined. The sizes of the cleavage products indicate that the enzyme cuts this substrate at the same sequences as double-stranded DNA (GANTC). To determine whether or not the recognition sites in single-stranded DNA have to be present in double stranded form in order to be cleaved, DNA fragments containing complementary or non-complementary Hinf I sequences were prepared and treated as substrates. The results suggest that completely base-paired recognition sites are necessary for cleavage. Sequences surrounding the Hinf I pentanucleotides significantly modulate the reaction rates. PMID- 6285306 TI - Nucleotide sequence of a region of the mitochondrial genome of Aspergillus nidulans including the gene for ATPase subunit 6. AB - A 1500 bp fragment of the Aspergillus nidulans mitochondrial genome contains genes for arginine and asparagine tRNAs, an unassigned reading frame, and the structural gene for ATPase subunit 6. The tRNA genes possess 66% nucleotide homology and possibly originated by a relatively recent duplication event. The unassigned reading frame displays a low level of homology with the human URF A6L. The predicted amino acid sequence of the A-nidulans ATPase subunit 6 gene is 40% homologous to the yeast polypeptide and includes the short, highly conserved regions also present in the equivalent subunits from other mitochondrial systems and from Escherichia coli. PMID- 6285308 TI - Effect of histone acetylation on the formation and removal of B(a)P chromatin adducts. AB - The modification of core histone proteins in mouse 10T1/2 cells and human lung epitheloid (A549) cells by B(a)PDE in vivo and in vitro was found to be similar. Only histones H2A and H3 were extensively modified. Also other proteins, possibly A24 protein and the minor histone H1 species seem to be binding relatively high levels of this ultimate carcinogen. Butyrate treatment which causes hyperacetylation of the core histones, did not change the specificity of B(a)PDE binding to core histones, nor did it affect the initial level of DNA modification. The acetylated species of histone H3 were all accessible to B(a)PDE, suggesting that these epsilon-amino-groups of the lysine residues are not the targets of the B(a)PDE. The rate of removal of B(a)P-DNA adducts was not affected by butyrate treatment in either normal human or XP fibroblasts. Furthermore the B(a)P-core histones were not preferentially removed from normal human fibroblast chromatin during a 24 h post-treatment incubation. PMID- 6285309 TI - Isolation and characterisation of the Xenopus laevis albumin genes: loss of 74K albumin gene sequences by library amplification. AB - The blood of the frog X.laevis contains 2 albumins of 68,000 and 74,000 daltons which are encoded in the liver by two related mRNAs. When an amplified X.laevis DNA library was screened with cloned albumin cDNA only 68,000 dalton albumin gene sequences were isolated. Hybridisation of the albumin cDNA to Southern-blots of Eco R1 digested X.laevis DNA showed that the sequences present in the recombinants did not account for all the fragments which hybridised on the Southern-blots. This indicated that 74K albumin gene sequences exist but that they are not present in the amplified DNA library. A X.laevis genomic library was therefore constructed and screened for albumin genes without amplification. Both 68K and 74K albumin gene sequences were isolated. Recombinants containing 74K albumin gene sequences grew extremely poorly and this probably explains why the 74K albumin sequences were ot isolated from the amplified library. Characterisation of the cloned DNA indicates that there is one sequence coding for the 68K albumin but two different sequences coding for the 75K albumin. PMID- 6285310 TI - The intervening sequence of the ribosomal RNA gene is highly conserved between two Tetrahymena species. AB - The entire intervening sequence of Tetrahymena thermophila ribosomal DNA has been determined. It is 413 nucleotides long and has the same splice junctions as those in T. pigmentosa. There is 93% homology between the intervening sequences in the two species, and 100% homology between their adjacent 26S RNA coding regions. PMID- 6285312 TI - The 5.8S RNA gene sequence and the ribosomal repeat of Schizosaccharomyces pombe. AB - We have characterized the rRNA gene repeat in Schizosaccharomyces pombe. This repeat, which does not contain the 5S RNA gene, is found in a 10.4 kb HindIII DNA fragment. We have determined the nucleotide sequences of the S. pombe 5.8S RNA gene and intergenic spacers from two different 10.4 kb DNA fragments. Analysis of isolated total cellular 5.8S RNA revealed the presence of eight species of 5.8S RNA, differing in the number of nucleotides at the 5'-end. The eight 4.8S RNA species vary in length from 158 to 165 nucleotides. Apart from the heterogeneity observed at the 5'-end, the sequence of the eight 5.8S RNA species appears to be identical and is the same sequence as coded for by the 5.8S genes. The gene sequence shows great homology to the 5.8S RNA genes or S. cerevisiae and N. crassa. Most of the base differences are confined to the highly variable stem though to be involved in co-axial helix stacking with the 25S RNA, where base pairing is nearly identical despite the sequence differences. Secondary structure models are examined in light of 5.8S RNA oligonucleotide conservation across species from yeasts to higher eukaryotes. PMID- 6285311 TI - Identification of a site of psoralen crosslinking in E. coli 16S ribosomal RNA. AB - We have developed a 2-dimensional gel method for identification of RNA sequences crosslinked by the intercalative drug 4'-hydroxymethyl-4,5',8-trimethylpsoralen (HMT). This method is being used to localize such sites in E. coli ribosomal RNA. We report here the identification of a site for HMT crosslinking within positions 434 and 497 of 16S rRNA. We suggest a likely site for HMT intercalation, in which residues U548 and U473 become crosslinked via the drug. PMID- 6285313 TI - Retrovirus-related sequences in human DNA: detection and cloning of sequences which hybridize with the long terminal repeat of baboon endogenous virus. AB - Human DNA sequences which hybridized with the long terminal repeats (LTR) of baboon type C virus M7 were detected by non-stringent blot hybridization. About 7 to 10 discrete bands of the LTR-related sequences were commonly observed in the DNAs from four independent human cell lines after digestion with either Eco RI, Hind III or Bam HI. The amounts of these sequences were more abundant in tumor cell lines than in a non-malignant cell line. The human sequences related to the M7 LTR seemed to be located at relatively specific sites on the cell DNA. The human DNA clones which hybridized with M7 LTR were detected in the human DNA library described by Lawn et al. (Cell 15, 1157-1174, 1978), at a frequency of about 300 per haploid genome. Five clones were isolated which shared different extent of homology with M7 LTR and whose restriction maps were totally different one another. The DNA structures of two of them resembled the genome of retroviruses. These results suggest the presence of various types of the LTR related sequences in human DNA: some of them might represent endogenous virus genomes of human cells. PMID- 6285315 TI - The effect of the delta subunit on the interaction of Bacillus subtilis RNA polymerase with bases in a SP82 early gene promoter. AB - We have examined the effect of the delta subunit on the interaction of the Bacillus subtilis RNA polymerase with an early gene promotor of phase SP82. Methylation by dimethyl sulfate, used to probe close approaches of polymerase to purines, revealed that noninitiated complexes formed by holo-enzyme (core-sigma delta) had significantly fewer contacts than complexes formed by core-sigma. The presence or absence of delta had little or no effect on close approaches to purines in initiated complexes. DNAase I footprinting indicated that core-sigma was bound to the same region regardless of whether delta, initiating nucleotides, or both, were present. These data support the conclusion that delta acts prior to initiation to enhance promoter selectivity by limiting the number of possible interactions that the polymerase can make with DNA. PMID- 6285314 TI - Organization of delta-crystallin genes in the chicken. AB - Double-stranded DNA was synthesized from delta-crystallin mRNA prepared from lens fibers of 15-day-old chick embryos and cloned at the Pst I site of the plasmid pBR322. Using the cloned cDNA and single-stranded cDNA as hybridization probes, a number of genomic DNA fragments containing delta-crystallin gene sequences have been cloned from the partial and complete EcoRI digests of chick brain DNA. One of the clones from the partial digests contains a DNA fragment that consists of four EcoRI fragments of 7.6 kb, 4.0 kb, 2.6 kb, and 0.8 kb. The gene sequences reside in the (5')7.6 kb - 0.8 kb - 4.0 kb (3') fragments. Electron microscopy has provided evidence that the cloned DNA fragment includes the entire gene sequences complementary to delta-crystallin mRNA except for the 3' terminal poly(A) tail, and that the delta-crystallin gene is interrupted by at least 13 intervening sequences. Another clone contains a genomic fragment that consists of two EcoRI fragments of 3.0 kb and 11 kb. The DNA fragment in the latter clone represents a different delta-crystallin gene, as judged by restriction endonuclease mapping and by electron microscopy. PMID- 6285316 TI - Antibody to poly(adenosine diphosphate-ribose) polymerase and its use in chromatin analysis. AB - To facilitate investigations on the organization of poly (ADP-Rib) polymerase in chromatin, and to elucidate its biological function, polymerase purified from HeLa nuclei was used to elicit antibodies in mice. The anti-polymerase sera was found to be specific by multiple criteria. The association of polymerase with oligonucleosomes of differing chain size was determined by the specific binding of polymerase antibody (and as control, anti-histone H3) to nitrocellulose transfers of native electrophoretic gels of these particles. By this technique, polymerase appears to be associated with a select subclass of nucleosomes. Further resolution of the polymerase binding sites within nucleosome classes was achieved with the antibody by two-dimensional native polyacrylamide gel electrophoresis and nitrocellulose transfer. PMID- 6285317 TI - In vitro potentiation of the activity of synthetic ovine corticotropin-releasing factor by arginine vasopressin. AB - The ability of arginine vasopressin (AVP) to potentiate the actions of synthetic ovine corticotropin-releasing factor (CRF) was examined using anterior pituitary fragments. Marked potentiation of ACTH release was observed upon incubating the fragments with a combination of 2 nM AVP and 1 nM CRF. Potentiation of CRF induced ACTH release was also observed when the fragments were incubated with a combination of 1 nM AVP and 0.5 nM CRF. These results suggest that AVP may play a role in the release of ACTH from the adenohypophysis. PMID- 6285320 TI - Effects of conditioned fear and environmental novelty on plasma beta-endorphin in the rat. AB - The levels of rat plasma beta-endorphin-like immunoreactivity following a 30 min exposure to (1) an unfamiliar operant chamber, (2) an unfamiliar operant chamber providing a VI-5 second schedule of 3 mA, 1 sec footshocks, or (3) a familiar chamber providing no shocks, but previously paired with unavoidable shocks were compared to control values from animals left undisturbed in their familiar home cages. The shocked group showed ten-fold elevations of beta-endorphin-like immunoreactivity compared to the undisturbed control animals, while conditioned rats showed a smaller two-fold elevation when re-exposed to a chamber in which they had previously been shocked. Two of five rats exposed merely to an unfamiliar chamber showed elevations, but there was no statistically reliable group effect. Such procedures may be useful for controlled and parametric studies of the mobilization of pituitary or brain pools of beta-endorphin under conditions involving merely anticipation of pain rather than the actual activation of ascending nervous pain pathways. PMID- 6285322 TI - Glomus jugulare tumor. AB - A 53-year-old woman with left vocal cord paralysis was seen for sharp shooting pains in the left side of the neck and the left shoulder and upper arm. Glomus jugulare tumor was diagnosed, based on clinical findings and results of angiography and computed tomography. Because surgery was contraindicated, radiotherapy was used. The patient's symptoms resolved within three months, and she had had no further problems. Glomus jugulare tumor should be considered in the differential diagnosis of adult chronic otitis, deafness, tinnitus, and dysfunction of the lower cranial nerves. PMID- 6285319 TI - Hypothalamic CRF-like immunoreactivity in the rat after hypophysectomy or adrenalectomy. AB - Hypothalamic CRF-like immunoreactivity was measured in normal, hypophysectomized or adrenalectomized adult male rats. As expected, adrenalectomy resulted in decreased levels in plasma corticosterone and increased plasma levels of ACTH; hypophysectomy resulted in decreased levels in both corticosterone and ACTH. The hypothalamic content of CRF-like immunoreactivity in animals two weeks post hypophysectomy or adrenalectomy was approximately seven times greater than that found in intact animals. At one week, post-surgery, small but statistically significant decreases in content of CRF-like immunoreactivity were observed. The results at one week are consistent with removal of feedback effects of ACTH and corticosterone causing increased release of CRF and decreased content. The increase in CRF-like immunoreactivity two weeks post-surgery is probably not related to direct feedback effects on release but may be due to increased synthesis secondary to long term removal of feedback inhibition. PMID- 6285321 TI - Epithelial and fibroblastoid cell lines derived from the normal canine prostate. I. Separation and characterization of epithelial and stromal components. AB - Cell cultures displaying exclusively epithelial or fibroblastoid morphology have been isolated by spillage and collagenase digest techniques, respectively. Primary cultures of both cell types have been readily subcultured. The use of a type I collagen substrate has been shown to be essential to the growth of normal prostatic epithelium in monolayer cultures. The ability to generate replicate subcultures of both cell types has allowed the quantitative characterization of the mitogenicity of fetal bovine serum and insulin in early subcultures. The control of culture conditions has permitted uniform cell population growth in early subculture with regular population doubling times in log phase of growth. Epithelial cultures have been shown to display many ultrastructural characteristics common to the normal epithelium of the canine prostate. PMID- 6285318 TI - Cholecystokinin octapeptide decreases intake of solid food in man. AB - Cholecystokinin octapeptide (CCK-OP) was reported to decrease the intake of liquid food in lean and in obese man. This study investigated the effect of CCK OP on the consumption of real life food, i.e., of standardized sandwiches. Sixteen young non-obese females and males participated, after an overnight fast, each in two experiments. After a basal 30 min, saline or CCK-OP, 1.5 or 3.0 Ivy Dog Units/kg body weight/15 min, was infused in random double blind fashion, while sandwiches were placed in front of the subjects. For the next three 15-min periods, the subjects were instructed to eat as much as they liked. In the first 15 min after 3.0 as well as 1.5 U CCK-OP/kg/15 min significantly fewer sandwiches (50 and 17 percent) were eaten than after saline (p less than 0.01 and p less than 0.05) and less hunger was reported (p less than 0.02 and p less than 0.05). Self-reported activation decreased only with 3.0 U CCK-OP (p less than 0.005). Reports of well-bring , electroencephalogram, heart rate, and respiration were not altered. The results support the notion that CCK is involved in the regulation of food intake. PMID- 6285323 TI - Nasal masses in children. PMID- 6285324 TI - Treatment of small cell lung carcinoma in association with cryptogenic fibrosing alveolitis. PMID- 6285327 TI - The science and art of healing. PMID- 6285326 TI - Heritabilities for regression and progression of rous sarcomas in the chicken. AB - Variance components for hatch, line, and sire expressed as percentages of the total variance for each parameter of regression and progression of Rous sarcomas in chickens were obtained. Some parameters measured specific stages of regression or progression, and other parameters measured the entire process of regression or progression of Rous sarcomas. Heritabilities of each parameter were estimated. Additive genetic variation is strongly implicated in the defense of the chicken against early tumor growth and during the entire process of regression. Environmental variation is important in influencing the point in time of tumor growth when the first reduction in size of tumor occurs. There is evidence of genetic variation exclusive of the R-Rs-1 gene locus that assists the chicken to resist tumor growth. The coefficient of variation for each of the parameters is very large as compared with the coefficient of variation for economic traits such as body weight at broiler age. PMID- 6285328 TI - The hand in practice. PMID- 6285325 TI - Major histocompatibility complex (B): effect on the response of chickens to a second challenge with rous sarcoma virus. AB - The influence of B genotype on the formation of a second RSV-induced tumor was studied. Line 6(3) (B2/B2) and (6(1) x 15(1))F3 and F4 B2/B2 and B5/B5 chickens were challenged with RSV in the left wingweb at 6 weeks of age. Tumor-bearing animals then were rechallenged with RSV in the right wingweb at either 10, 11, or 12 days post primary inoculation. Most B2/B2 hosts resisted development of a second tumor compared to none of the B5/B5 hosts. Failure of a second tumor to develop in B2/B2 hosts was not explained by humoral neutralization of RSV in vivo, since sera from 83% of B2/B2 and 100% of B5/B5 tumor bearing chickens failed to neutralize virus prior to 12 days post primary inoculation. Humoral antiviral immunity appeared later in B5/B5 than in B2/B2 hosts. Humoral antiviral immunity appears to play little if any role in resistance to early RSV-induced tumor development, but likely both humoral and cell-mediated antitumor immunity do. PMID- 6285329 TI - [Bronchogenic carcinoma--pathoanatomy. (Studies of 200 resection specimen)]. PMID- 6285330 TI - [The role of cytology in the diagnosis of bronchogenic carcinoma (author's transl)]. PMID- 6285331 TI - [Specific activity and molecular forms of NAD-kinase in Neurospora crassa ontogeny]. AB - The specific activity and molecular forms of NAD-kinase during ontogenesis of Neurospora crassa were investigated. The specific activity of the enzyme increased drastically at critical stages of fungal development, i.e. during conidia germination and during transition from the logarithmic to stationary growth stage, reaching 85 nmole NADP/hr/mg protein. By polyacrylamide gel electrophoresis four forms of NAD-kinase were revealed that had the following molecular masses: I-338,000, II-306,000, III-229,000, and IV-203,000. The vegetative mycelium contained predominantly form III, and conidia showed a high content of high-molecular-weight forms I and II. PMID- 6285332 TI - [Isolation and properties of an alpha-galactosidase preparation from Cephalosporium sp. 237]. AB - Different methods for preparing alpha-galactosidase from the culture fluid of the micromycete Cephalosporium sp. 237 were tested. They included precipitation with ethanol, isopropanol, and acetone. Precipitation with two volumes of acetone gave the best results with respect to specific activity (12 units) and yield of the enzyme (78%). Properties of alpha-galactosidase (optimum pH at 5.5, temperature optimum at 40 degrees C. thermolability, etc), when different substrates were used, were examined. PMID- 6285333 TI - [Changes in the function of the hypothalamo-hypophyseo-adrenal system and the thymus as affected by corticotropin in guinea pigs]. AB - Functional state of the hypothalamic-hypophyseal-adrenal system and the thymus was studied in thymic hypo- and hyperplasia of guinea pigs. It was detected that thymomegalia is accompanied by STH and prolactin hyperproduction in adeno hypophysis, intensified lymphopoiesis (at the expense of T-lymphocytes) and the adrenal glucocorticoid function inhibition, resultant of the lowered ACTH secretion, as well as by the enhanced catecholamine and serotonin content in the hypothalamus. Reverse changes are seen in thymic hypoplasia following intramuscular ACTH injection in a dose 1 U/kg/day within 5 to 10 days. PMID- 6285334 TI - [Nature of the intracellular thyroid hormone receptor]. AB - Thyroxine-binding proteins were isolated and purified by means of affine chromatography from the blood serum, 105 000 g of cytosol supernatant, non histone protein fractions of chromatin, extracted with 0.4 M KCl. Identity of their molecular weights and of end aminoacids was found. A comparative study of proteins above, using chromatoelectrophoresis, has shown identity of their primary structure. The data obtained indicate, that chromatin and cytosol contain universal, thyroxine-binding protein, which structure is identical with that of prealbumin thyroxine-binding protein. This protein is considered to be a receptor, realizing the control of genome expression by thyroid hormones. PMID- 6285335 TI - [Action of thyroid hormones and somatotropin on the free radical, iron-sulfur protein and cytochrome P-450 state in the liver]. AB - The state of free radicals, iron-sulphuric proteins and cytochrome P-450 was determined in hypo- and hyperthyroid rats by means of EPR method. It was shown that EPR signal of cytochrome P-450 was increased in the liver of thyroidectomized rats, characterized by somatotropin deficiency and delayed metabolism. It decreased under the action of exogenous somatotropin both in intact and in thyroidectomized animals. Cytochrome P-450 was lowered in hyperthyroid rats, characterized by intensified oxidative processes without signs of the enhanced hepatic growth. The feedback between the intensified hepatic oxidative processes and the activity of cytochrome P-450-dependent multipurposeful oxidases is suggested. PMID- 6285336 TI - [Effect of human serum lipoproteins on various indices of thyroid function in rats]. AB - It was shown that 7 to 10 days' injection of lipoproteins (LPs), isolated from the human blood serum according to Burstein and Scholnick method, induces significant inhibition of the rat thyroid iodine-absorbing function. Alpha-LPs exert more evident inhibiting effect than beta + pre- beta-LPs, and beta + pre- beta-LPs from the oncological patients' blood serum have more pronounced inhibiting effect comparatively to the same fraction of the donors; blood serum. 10-days' LP injection is accompanied by thyroxine inhibition and iodothyrozine/iodothyronine ratio rise in the thyroid tissue. In the experiments in vitro and in vivo several Lp fractions decreased thyrotropin-stimulating effect on AMP accumulation in the rat thyroid gland. Possible mechanisms of LP action on the thyroid function are discussed. The study of LP effect on the thyroid function in homologous system is necessary. It is suggested that hyperlipidemia (hyperlipoproteinemia) is one of the factors, contributing to the thyroid function decrease with age and in patients with malignant tumors. PMID- 6285337 TI - [Adaptation problems and solar activity]. PMID- 6285338 TI - Embryonic brain extract induces collagen biosynthesis in cultured muscle cells: involvement in acetylcholine receptor aggregation. AB - The involvement of extracellular matrix components in induction of the aggregation of acetylcholine (AcCho) receptors by factor(s) present in embryonic brain extract was investigated. Embryonic brain extract induced a three-fold increase in the number of AcCho receptor aggregates on the surface of cultured myotubes and a 5- to 10-fold increase in the synthesis of procollagen, which was secreted into the medium and converted to collagen. Adult brain extract, embryonic serum, and embryonic liver extract were less active in stimulating both collagen synthesis and AcCho receptor aggregation. A physiological connection between the two processes is suggested, since the number of AcCho receptor aggregates could be reduced to control levels by treating brain extract stimulated myotubes with purified bacterial collagenase. In addition, stimulation of collagen secretion by ascorbic acid (50 micrograms/ml) promoted a 1.6-fold increase in AcCho receptor aggregation. When ascorbic acid was added together with the brain extract, further increases in both collagen synthesis and AcCho receptor aggregation were observed. PMID- 6285339 TI - Anthrax toxin edema factor: a bacterial adenylate cyclase that increases cyclic AMP concentrations of eukaryotic cells. AB - Anthrax toxin is composed of three proteins: protective antigen (PA), lethal factor (LF), and edema factor (EF). These proteins individually cause no known physiological effects in animals but in pairs produce two toxic actions. Injection of PA with LF causes death of rats in 60 min, whereas PA with EF causes edema in the skin of rabbits and guinea pigs. The mechanisms of action of these proteins have not been determined. It is shown here that EF is an adenylate cyclase [ATP pyrophosphate-lyase (cyclizing), EC 4.6.1.1] produced by Bacillus anthracis in an inactive form. Activation occurs upon contact with a heat-stable eukaryotic cell material. The specific activity of the resulting adenylate cyclase nearly equals that of the most active known cyclase. In Chinese hamster ovary cells exposed to PA and EF, cAMP concentrations increase without a lag to values about 200-fold above normal, remain high in the continued presence of toxin, and decrease rapidly after its removal. The increase in cAMP is completely blocked by excess LF. It is suggested that PA interacts with cells to form a receptor system by which EF and perhaps LF gain access to the cytoplasm. PMID- 6285340 TI - Nucleotide sequence of the transforming gene of simian sarcoma virus. AB - The sequence of the transforming region of simian sarcoma virus (SSV) has been determined by using molecularly cloned viral DNA. This region encompassed the 1.0 kilobase pair woolly monkey cell-derived insertion sequence, v-sis, and flanking simian sarcoma-associated viral (SSAV) sequences. A 675-nucleotide-long open reading frame commenced 19 nucleotides within the SSAV sequences to the left of the v-sis helper viral junction and terminated within v-sis itself. Possible promoter and acceptor splice signals were detected in helper viral sequences upstream from this open reading frame, and potential polyadenylylation sites were identified downstream both within v-sis and in helper viral sequences beyond v sis. The recombinational event that led to the generation of SSV occurred in the middle of two functional codons, indicating that SSAV provided the regulatory elements for transcription as well as the initiation codon for translation of SSV cell-derived transforming sequences. PMID- 6285341 TI - Sodium channels induced by depolarization of the Xenopus laevis oocyte. AB - An electrically gated Na+ channel can be made to appear in the membrane of the Xenopus laevis oocyte by simple depolarization. This membrane normally responds passively to imposed transmembrane currents with resting potentials around -60 mV, but when it is held depolarized to more than about +30 mV it becomes possible to obtain long-lasting regenerative depolarizations up to +80 mV; these depolarizations can last as long as 20 min. This potential is due to an "induction" of a Na+-dependent channel that is electrically gated open and closed. Its threshold for opening is about -20 mV and it is selective for Na+ over Cs+ and choline+ but is blocked by relatively small quantities of Li+. When a long voltage clamp step to a positive potential under ENa (+70 to +90 mV) is applied, an inward current is observed for many minutes, implying that this channel does not have an inactivation mechanism. The inward Na+ current is blocked by 0.50 mM tetrodotoxin. When the membrane is held at or near resting potential, the excitability will disappear with time, but it can be made to reappear by again depolarizing the membrane. PMID- 6285342 TI - cAMP and spore differentiation in Dictyostelium discoideum. AB - The individual amoebae composing of Dictyostelium aggregate can differentiate into either stalk or spore cells according, it is believed, to the extracellular signals they receive. By inducing the differentiation of isolated cells we hope to identify these signals. It is shown here that wild-type cells can differentiate into prespore cells in a solution of cAMP plus salts supplemented by conditioned medium. Cell-to-cell contact is not required. More important, isolated cells of various sporogenous mutant strains form spores in similar conditions without needing conditioned medium at all. For these strains at least, cAMP is the sole inducer necessary for spore formation. Earlier work has shown that stalk cells are induced by a combination of cAMP and a low molecular weight factor, differentiation inducing factor (DIF). DIF now appears to be the only pathway-specific inducer involved in the differentiation of sporogenous amoebae and DIF levels in the aggregate may therefore determine whether an amoeba becomes a stalk or a spore cell. In suitable conditions some sporogenous mutants form migrating slugs having an anterior/posterior pattern of prestalk and prespore cells. This pattern could be generated by the localized activity of DIF. PMID- 6285343 TI - Human transferrin receptor: expression of the receptor is assigned to chromosome 3. AB - Human chromosome 3 has been identified as responsible for expression of the transferrin receptor in mouse-human lymphocyte hybrids. The receptor was detected by immunoprecipitation with anti-human receptor antibody of 125I-labeled cells. This method also detected a similar 94,000-dalton protein in mouse cells. A radioimmunoassay developed for the human transferrin receptor measured 10% crossreactivity with the mouse protein. The two proteins were distinguished by NaDodSO4/polyacrylamide gel patterns of partial proteolytic digests of the immunoprecipitated proteins. Mouse-human hybrids were generated by fusing a mouse thymoma (BW5147) cell line to either concanavalin A- or pokeweed mitogen activated human peripheral blood lymphocytes or a mouse myeloma (NS-1) to uncultured human peripheral blood lymphocytes. Each hybrid was karyotyped with respect to both mouse and human chromosomes. In every case, expression of the human transferrin receptor correlated only with human chromosome 3. PMID- 6285344 TI - Novel features of animal mtDNA evolution as shown by sequences of two rat cytochrome oxidase subunit II genes. AB - The sequence of the region of the mitochondrial genome that encodes cytochrome oxidase subunit II (COII) has been determined for each of two closely related rat species, Rattus norvegicus and R. rattus. Comparison of the two sequences shows that 94.4% of the nucleotide substitutions are silent. The occurrence of this high proportion of silent substitutions leads us to propose that the rapid evolution of mtDNA relative to nuclear DNA is due only to silent changes and that amino acid-altering substitutions accumulate in nuclear and mtDNA at comparable rates. Other novel features of the nucleotide substitution pattern in the rat COII gene are a high transition/transversion ratio (8.0:1) and a strong bias toward C in equilibrium T transitions in the light strand. Comparison of the R. norvegicus COII sequence with the bovine and human sequences show that there may be selective constraints on some silent positions within the gene and that its rate of evolution may be different in different mammalian lineages. PMID- 6285345 TI - Impaired receptor-mediated catabolism of low density lipoprotein in the WHHL rabbit, an animal model of familial hypercholesterolemia. AB - The homozygous WHHL (Watanabe heritable hyperlipidemic) rabbit displays either no or only minimal low density lipoprotein (LDL) receptor activity on cultured fibroblasts and liver membranes and has therefore been proposed as an animal model for human familial hypercholesterolemia. To assess the impact of this mutation on LDL metabolism in vivo, we performed lipoprotein turnover studies in normal and WHHL rabbits using both native rabbit LDL and chemically modified LDL (i.e., methyl-LDL) that does not bind to LDL receptors. The total fractional catabolic rate (FCR) for LDL in the normal rabbit was 3.5-fold greater than in the WHHL rabbit. Sixty-seven percent of the total FCR for LDL in the normal rabbit was due to LDL receptor-mediated clearance and 33% was attributable to receptor-independent processes; in the WHHL rabbit, essentially all of the LDL was catabolized via receptor-independent processes. Despite a 17.5-fold elevated plasma pool size of LDL apoprotein (apo-LDL) in WHHL as compared to normal rabbits, the receptor-independent FCR-as judged by the turnover of methyl-LDL-was similar in the two strains. Thus, the receptor-independent catabolic processes are not influenced by the mutation affecting the LDL receptor. The WHHL rabbits also exhibited a 5.6-fold increase in the absolute rate of apo-LDL synthesis and catabolism. In absolute terms, the WHHL rabbit cleared 19-fold more apo-LDL via receptor-independent processes than did the normal rabbit and cleared virtually none by the receptor-dependent pathway. These results indicate that the homozygous WHHL rabbit shares a number of metabolic features in common with human familial hypercholesterolemia and should serve as a useful model for the study of altered lipoprotein metabolism associated with receptor abnormalities. We also noted that the in vivo metabolic behavior of human and rabbit LDL in the normal rabbit differed such that the mean total FCR for human LDL was only 64% of the mean total FCR for rabbit LDL, whereas human and rabbit methyl-LDL were cleared at identical rates. Thus, if human LDL and methyl-LDL had been used in these studies, the magnitude of both the total and receptor-dependent FCR would have been underestimated. PMID- 6285346 TI - Identification of an antigen associated with transforming genes of human and mouse mammary carcinomas. AB - Sera from tumor-bearing mice immunoprecipitated a 86,000-dalton glycoprotein from extracts of NIH cells transformed by human mammary carcinoma DNA. This antigen was not immunoprecipitated from extracts of NIH 3T3 cells, spontaneously transformed NIH cells, NIH cells transformed by normal human DNA, NIH cells transformed by human bladder carcinoma DNA, or NIH cells transformed by Rous sarcoma virus DNA. In addition, sera from mice bearing tumors induced by NIH cells transformed by either normal human DNA or human bladder carcinoma DNA did not immunoprecipitate this antigen from extracts of NIH cells transformed by human mammary carcinoma DNA. However, this antigen was immunoprecipitated by sera from mice bearing tumors induced by NIH cells transformed by mouse mammary carcinoma DNAs and from mice bearing primary mammary carcinomas. These results indicate that this glycoprotein represents an antigen that is specifically associated with expression of the transmissible transforming genes of human and mouse mammary carcinomas. PMID- 6285347 TI - Interaction of the Escherichia coli dnaA initiation protein with the dnaZ polymerization protein in vivo. AB - To define in vivo interactions of Escherichia coli DNA replication components, extragenic suppressors of a dnaZ(TS) mutant were isolated. A temperature sensitive dnaZ mutant, which is defective in polymerization, was placed at 39 degrees C to select temperature-insensitive revertants. Some of these revertants also were cold sensitive, a phenotypic property that facilitated study of the suppressor. Mapping of the cold sensitivity indicated that some of the suppressor mutations are intragenic but others are located within the initiation gene, dnaA. The dnaA mutations that suppress the dnaZ(TS) defect are designated dnaA(SUZ, CS). The dnaA(SUZ, CS) strains have a defect in DNA synthesis at low temperature that is typical of an initiation defect. These data suggest that the dnaA product, an initiation factor, interacts in vivo with the dnaZ protein, a polymerization factor. PMID- 6285348 TI - Human papilloma viral DNA replicates as a stable episome in cultured epidermal keratinocytes. AB - Human papilloma virus (HPV) is poorly understood because systems for its growth in tissue culture have not been developed. We report here that cultured human epidermal keratinocytes could be infected with HPV from plantar warts and that the viral DNA persisted and replicated as a stable episome. There were 50-200 copies of viral DNA per cell and there was no evidence to indicate integration of viral DNA into the cellular genome. There was also no evidence to suggest that viral DNA underwent productive replication. We conclude that cultured human epidermal keratinocytes may be a model for the study of certain aspects of HPV biology. PMID- 6285349 TI - Functional exchange of components between light-activated photoreceptor phosphodiesterase and hormone-activated adenylate cyclase systems. AB - Previous studies have noted profound similarities between the regulation of light activated 3',5'-cyclic nucleotide phosphodiesterase (3',5'-cyclic-nucleotide 5' nucleotidohydrolase, EC 3.1.4.17) in retinal rods and hormone-activated adenylate cyclase [ATP pyrophosphate-lyase (cyclizing), EC 4.6.1.1] in a variety of tissues. We report here the functional exchange of components isolated from the photoreceptor system, which displayed predicted functional characteristics when incubated with recipient adenylate cyclase systems from rat cerebral cortical and hypothalamic synaptic membranes and frog erythrocyte ghosts. We demonstrate functional exchange of photoreceptor components at each of three loci: the hormone receptor, the GTP-binding protein (GBP), and the catalytic moiety of adenylate cyclase. Illuminated (but not unilluminated) rhodopsin was fund to mimic the hormone-receptor complex, causing GTP-dependent activation of adenylate cyclase. The photoreceptor GBP complexed with guanosine 5'-[beta, gamma)imidotriphosphate (p[NH]ppG) produced a marked activation of recipient adenylate cyclase systems. Much smaller activation was observed when GBP was not complexed with p[NH]ppG. A heat-stable photoreceptor phosphodiesterase inhibitor reduced both basal and Mn2+-activated adenylate cyclase activities and this inhibition was reversed by photoreceptor GBP.p[NH]ppG. These data demonstrate a remarkable functional compatibility between subunits of both systems and furthermore imply that specialized peptide domains responsible for protein protein interactions are highly conserved. PMID- 6285351 TI - Bacteriophage lambda DNA packaging: scanning for the terminal cohesive end site during packaging. AB - Bacteriophage lambda packages the DNA of the related phage 21 poorly [Hohn, B. (1975) J. Mol. Biol. 98, 93--106]. To understand the nature of the packaging defect, the interaction of the cohesive end site (cos) specific for phage 21 (cos phi 21) with phage lambda terminase has been investigated. The ability of lambda terminase to cleave cos phi 21 was studied in vitro; lambda terminase cleaved cos phi 21 only 1% as well as it cleaved the phage lambda cohesive end site (cos lambda). In vitro packaging experiments showed that the lambda and 21 packaging specificities observed in vivo are also found in vitro. The cos cleavage reaction was modified so that competition experiments could be performed; these experiments showed that cos phi 21 was unable to bind lambda terminase, thus identifying the nature of the defect. Previous work [Feiss, M., Fisher, R. A., Siegele, D. A., Nichols, B. P. & Donelson, J. E. (1979) Virology 92, 56--67] has shown that the base pairs giving lambda or 21 packaging specificity are at the left end of the chromosome, outside the 22-base-pair symmetry region that includes the annealed cohesive ends. Therefore, terminase binding to cos requires interactions with base pairs to the Nu1 side of the cohesive end symmetry segment. The evidence supports the proposition that cos consists of adjacent sites for binding of terminase and for nicking by terminase. Because cos phi 21 can be cut by lambda terminase to terminate DNA packaging, it is proposed that the terminase that binds and nicks at the initial cos site is brought into contact with the terminal cos site by the packaging process. Terminase recognizes and nicks the cohesive end sequence of the terminal cos without requiring the binding site. PMID- 6285350 TI - Stimulation of cyclic AMP accumulation and corticotropin release by synthetic ovine corticotropin-releasing factor in rat anterior pituitary cells: site of glucocorticoid action. AB - A 2.5-fold stimulation of cyclic AMP cellular content is measured 60 sec after addition of 100 nM synthetic ovine corticotropin-releasing factor (C-RF; corticoliberin) to rat anterior pituitary cells in culture. A maximal response of cyclic AMP content at 400% above control is observed between 2 and 30 min after addition of the peptide, whereas an 8-fold stimulation of cyclic AMP released into the incubation medium is measured between 10 and 180 min. A linear 7-fold increase of corticotropin release is observed for up to 3 hr. Preincubation from 18 hr with the potent glucocorticoid dexamethasone has no effect on C-RF-induced cyclic AMP accumulation. The same treatment with dexamethasone causes an 80% inhibition of corticotropin release induced by both C-RF and the cyclic AMP derivative 8-bromoadenosine 3',5'-cyclic monophosphate. The present data show that ovine C-RF is a potent stimulator of cyclic AMP accumulation in rat anterior pituitary cells and that the process is insensitive to the action of dexamethasone. The marked inhibition by dexamethasone of corticotropin secretion induced by a cyclic AMP derivative indicates that glucocorticoids exert their potent inhibitory effects on corticotropin secretion at a step distant to cyclic AMP formation. PMID- 6285352 TI - Estimation of phylogenetic relationships from DNA restriction patterns and selection of endonuclease cleavage sites. AB - The distribution of cleavage sites and their related sequences have been analyzed for 54 restriction endonucleases in the genome of human mitochondrial DNA; in three papova viruses, BK, simian virus 40, and polyoma; and in three bacteriophages, phi X174, fd, and G4. The results show that the cleavage sites and related sequences for most of the restriction enzymes tested are distributed nonrandomly. These results (i) constitute prima facie evidence for the action of natural selection, either direct or indirect on the restriction sites, and (ii) suggest that estimates of phylogenetic relationship, based on a phenetic approach using restriction enzyme data, will be biased. PMID- 6285353 TI - Hepatic uptake of chylomicron remnants in WHHL rabbits: a mechanism genetically distinct from the low density lipoprotein receptor. AB - Homozygous Watanabe hereditary hyperlipidemic (WHHL) rabbits have a near-complete deficiency of low density lipoprotein (LDL) receptors in liver and other tissues. As a result, these rabbits clear LDL from plasma at an abnormally slow rate. In the current studies we show that WHHL rabbits clear chylomicrons from plasma at a normal rate. Chylomicrons are cleared by a two-step process: (i) hydrolysis of triglycerides in extrahepatic tissues to yield cholesteryl ester-rich remnant particles and (ii) rapid uptake of the remnants by liver. Normal and WHHL rabbits were given intravenous injections of rat chylomicrons labeled either in the lipid portion with [3H]cholesterol and [14C]palmitate or in the protein portion with [125]iodine. All radiolabeled components were removed from plasma at comparable rates in normal and WHHL rabbits. Comparable amounts of radioactivity accumulated in livers of animals from both genotypes. In vitro assays showed that liver membranes from WHHL rabbits were markedly deficient in the binding of 125I labeled chylomicron remnants as well as 125I-labeled LDL, implying that chylomicron remnants can bind to the hepatic LDL receptor. We conclude that the rabbit liver normally has at least two genetically distinct lipoprotein uptake mechanisms, both of which recognize chylomicron remnants: (i) the LDL receptor and (ii) a specific chylomicron remnant uptake mechanism that is not measured adequately by current in vitro membrane binding assays. WHHL rabbits possess a normal chylomicron remnant uptake mechanism that allows them to clear chylomicrons from plasma at a rapid rate despite their genetic deficiency of LDL receptors. PMID- 6285354 TI - Use of restriction endonucleases for mapping the allele for beta s-globin. AB - We have reported the direct analysis of the allele for beta 2-globin by using restriction endonuclease Dde I coupled with blot-hybridization analysis. In that report we predicted that a major use of our analysis could be for the prenatal diagnosis of sickle cell anemia. Here we present such an analysis. In addition, this report also describes the use of a new enzyme Mst II, which also distinguish the beta s allele from the normal beta-globin allele. Blot-hybridization analysis with restriction endonuclease Mst II shows the 5' end of the normal beta-globin gene to reside on a fragment of approximately 1.14 kilobases, whereas the 5' end of the beta s-globin gene resides on a fragment of approximately 1.34 kilobases. Because the fragment sizes generated by Mst II are significantly larger than those generated by Dde I, one can easily perform a prenatal diagnosis for sickle cell by standard blot hybridizations onto nitrocellulose filters. PMID- 6285356 TI - Functional reconstitution of the purified sodium channel protein from rat sarcolemma. AB - The purified saxitoxin (STX) binding component of the rat sarcolemmal sodium channel (SBC) has been reconstituted into phospholipid vesicles. The reconstituted SBC displays the pharmacological properties and the ability to control sodium fluxes expected of a functional sodium channel. Batrachotoxin (BTX) increases 22Na+ influx into reconstituted SBC vesicles by greater than 100% over control at early time points. The BTX-stimulated 22Na+ influx is specifically and quantitatively blocked by STX. Veratridine and aconitine also stimulate Na+-flux--although less effectively than BTX--in the order: BTX greater than veratridine greater than aconitine. The logarithmic dose--response curves for BTX and veratridine are sigmoidal with a K0.5 of 1.5 microM and 35 microM, respectively. Vesicles containing the reconstituted SBC demonstrate 3H-labeled STX binding to a single class of high affinity sites witha Kd of 5--7 nM at 0 degrees C; the thermal stability of the STX receptor is markedly enhanced by reconstitution. Our results confirm that the purified STX binding component from rat sarcolemma constitutes the sodium channel itself and contains at least those components sufficient for channel activation, transmembrane ion movement, and inhibition by STX. PMID- 6285355 TI - Transforming genes of human bladder and lung carcinoma cell lines are homologous to the ras genes of Harvey and Kirsten sarcoma viruses. AB - Blot hybridization analysis indicated that NIH 3T3 mouse bladder transformed by high molecular weight DNAs of a human bladder and a human lung carcinoma cell line contained new sequences homologous, respectively, to the transforming genes of Harvey (rasH) and Kirsten (rasK) sarcoma viruses. The unique ras sequences were present in multiple independent NIH cell lines transformed in both primary and secondary transfection assays and corresponded to ras sequences normally present in human DNAs. The ras gene product was expressed in NIH cells transformed by bladder carcinoma DNAs and in the human bladder carcinoma cell lines at levels 2- to 4-fold greater than the level observed in nontransformed NIH 3T3 cells. These results indicate that the transforming genes of these human tumor cell lines are the cellular homologs of two retroviral transforming genes. PMID- 6285357 TI - [3H]nitrendipine-labeled calcium channels discriminate inorganic calcium agonists and antagonists. AB - [3H]Nitrendipine binds with high affinity to brain membranes with a drug specificity indicating association with sites mediating the pharmacologic actions of dihydropyridine slow-calcium-channel antagonist drugs. In brain membranes, [3H]nitrendipine binding is absolutely dependent on the presence of calcium ions. Interactions of cation with [3H]nitrendipine binding sites correlate with their physiologic actions at voltage-dependent calcium channels. Ions such as strontium and barium, which mimic calcium physiologically, share the action of calcium in enhancing [3H]nitrendipine binding. Ions such as lanthanum an cobalt, which block the effects of calcium, can inhibit [3H]nitrendipine binding and block the stimulating actions of calcium. The ability to monitor the influence of ions on an agonist-antagonist continuum at [3H]nitrendipine binding sites provides a molecular probe to explore the regulation of cellular function by calcium and other cations. PMID- 6285358 TI - Acute leukemia viruses E26 and avian myeloblastosis virus have related transformation-specific RNA sequences but different genetic structures, gene products, and oncogenic properties. AB - Replication-defective acute leukemia viruses E26 and myeloblastosis virus (AMV) cause distinct leukemias although they belong to the same subgroup of oncogenic avian tumor viruses based on shared transformation-specific (onc) RNA sequences. E26 causes predominantly erythroblastosis in chicken and in quail, whereas AMV induces a myeloid leukemia. However, upon cultivation in vitro for >1 month, a majority of surviving hemopoietic cells of E26-infected animals bear myeloid markers similar to those of AMV-transformed cells. We have analyzed the genetic structure and gene products of E26 virus for a comparison with those of AMV. An E26/helper virus complex was found to contain two RNA species: a 5.7-kilobase (kb) RNA that hybridizes with cloned AMV-specific proviral DNA and hence is probably the E26 genome; and an 8.5-kb RNA that is unrelated to AMV and represents helper virus RNA. Thus, E26 RNA is smaller than 7.5-kb AMV RNA. Hybridization of size-selected poly(A)-terminating E26 RNA fragments with AMV specific DNA indicated that the shared specific sequences are located in the 5' half of the E26 genome as opposed to a 3' location in AMV RNA. In nonproducer cells transformed in vitro by E26, a gag-related nonstructural 135,000-dalton protein (p135) was found. No gag(Pr76) or gag-pol (Pr180) precursors of essential virion proteins, which are present in AMV nonproducer cells, were observed. p135 was also found in cultured E26 virus producing cells of several leukemic chickens, and its intracellular concentration relative to that of the essential virion proteins encoded by the helper virus correlates with the ratio of E26 to helper RNA in virions released by these cells. p135 is phosphorylated but not glycosylated; antigenically it is not related to the pol or env gene products. It appears to be coded for by a partial gag gene and by E26-specific RNA sequences, presumably including those shared with AMV. Hence, AMV and E26 appear to use different strategies for the expression of related onc sequences: AMV is thought to encode a transforming protein via a subgenomic mRNA, whereas E26 codes for a gag-related polyprotein via genomic RNA. It is speculated that differences in the oncogenic properties of E26 and AMV are due to differences in their genetic structures and gene products. PMID- 6285359 TI - Alteration in phospholipid methylation and impairment of signal transmission in persistently paramyxovirus-infected C6 rat glioma cells. AB - The paramyxoviruses measles (subacute sclerosing panencephalitis, SSPE) virus and canine distemper virus (CDV) cause an impairment of the catecholamine-induced beta-adrenergic receptor-dependent cAMP generation in persistently infected C6 rat glioma cells. In C6 cells persistently infected with CDV the number of receptors is greatly reduced. Hirata and Axelrod have shown that the number of beta-adrenergic receptors could be regulated by methylation of phosphatidylethanolamine, resulting in lecithin synthesis [Hirata, F. & Axelrod, J. (1980) Science 209, 1082-1090]. We have therefore studied the methylation of phosphatidylethanolamine in persistently infected cells by the incorporation of [3H]methyl groups from [methyl-3H]methionine into phosphatidylethanolamine. In both infected systems, C6/ SSPE and C6/CDV, we observed a total loss of catecholamine-stimulated beta-adrenergic receptor-dependent methylation, whereas the beta-receptor-independent methylation of phospholipids was unchanged. PMID- 6285361 TI - Illegitimate recombination mediated in vitro by DNA gyrase of Escherichia coli: structure of recombinant DNA molecules. AB - We have developed a cell-free system from Escherichia coli for studying illegitimate recombination between nonhomologous DNA molecules. The recombination is stimulated by oxolinic acid, an inhibitor of DNA gyrase. The stimulation is abolished by coumermycin A1 and is not found in extracts of nalidixic acid resistant (gyrA) mutants. We therefore inferred that DNA gyrase directly participates in illegitimate recombination, at least in the presence of oxolinic acid [Ikeda, H., Moriya, K. & Matsumoto, T. (1981) Cold Spring Harbor Symp. Quant. Biol. 45, 399--408]. The structure of recombinant DNA molecules formed in the presence of oxolinic acid from a cross between phage lambda and plasmid pBR322 DNAs was analyzed by heteroduplex mapping. Among nine isolates tested, two recombinants were formed by the insertion of the plasmid into the lambda genome. The seven other recombinants had more complicated genome structures. Insertion of pBR322 was accompanied by a deletion on one of the genomes. In all cases, the end points of deletions coincided with one end of the pBR322 insertion. Recombination sites seemed to be distributed randomly on the lambda and pBR322 genomes. Analysis of nucleotide sequences of the recombination junctions proved that the crossover took place between nonhomologous DNA sequences. A model for DNA gyrase mediated illegitimate recombination is discussed. PMID- 6285360 TI - Reciprocal effects of an inhibitory factor on catalytic activity and noncatalytic cGMP binding sites of rod phosphodiesterase. AB - In illuminated rod outer segment membranes, GTP and guanosine 5'-[beta, gamma imido]triphosphate (p[NH]ppG) have reciprocal effects on cGMP phosphodiesterase (PDEase; 3':5'-cyclic-nucleotide 5'-nucleotidohydrolase, EC 3.1.4.17) activity and cGMP binding to noncatalytic sites on that enzyme. Two micromolar p[NH]ppG increased PDEase activity more than 2-fold while inhibiting cGMP binding more than 40%. Reduction of noncatalytic cGMP binding, which followed addition of p[NH]ppG, was not a result of PDEase activation. Both effects of p[NH]ppG were completely dependent on the presence of bleached rhodopsin. A heat-stable factor has been found to inhibit PDEase activity and also to stimulate cGMP binding to noncatalytic cGMP binding sites. Addition of p[NH]ppG reversed the effects of this factor on both PDEase activity and cGMP binding. During purification of this material, the activity peaks for both PDEase inhibition and activation of noncatalytic cGMP binding comigrated on both Blue Sepharose CL-6B column chromatography and sucrose density gradients centrifugation, suggesting that the same factor could be responsible for both inhibition of PDEase activity and enhancement of noncatalytic cGMP binding. Limited tryptic proteolysis of PDEase, which markedly reduced cGMP binding to the noncatalytic sites, and experiments using highly purified cAMP (free of cGMP) as substrate for PDEase showed that the binding of cGMP to noncatalytic sites was not required for the heat-stable inhibitory factor to inhibit PDEase activity. We discuss possible relationships between the regulation of PDEase and the binding of cGMP to noncatalytic sites. PMID- 6285362 TI - Covalent crosslinking of angiotensin II to its binding sites in rat adrenal membranes. AB - To identify the molecular components of the angiotensin II (AII) receptor in the adrenal cortex, we have covalently linked 125I-labeled AII (125I-AII) to its binding sites by using the chemical crosslinker disuccinimidyl suberate. Membrane fractions from rat adrenal tissue were incubated with 125I-AII, washed, and treated with disuccinimidyl suberate. NaDodSO4/polyacrylamide gel electrophoresis carried out under reducing conditions followed by autoradiography showed that a major polypeptide(s) with an average Mr of 116,000 was covalently tagged with 125I-AII. Proteins with Mr of 54,000 and 45,000 were also lightly labeled. Labeling of the Mr 116,000 species was specific in that it could be abolished by prior incubation of the membranes with [Sar1,Leu8]AII or unlabeled AII. Labeling of the smaller proteins was less affected by prior incubation with [Sar1,Leu8]AII. Experiments in which crosslinking was carried out in the presence of a variety of protease inhibitors indicated that the Mr 116,000 species is not an enzyme involved in AII degradation. Gel electrophoresis under nonreducing conditions showed that this component is not derived from larger disulfide-linked entities. We suggest that the Mr 116,000 moiety is a part of the receptor system for AII. The relationship, if any, of the Mr 54,000 and 45,000 peptides to the AII receptor remains unknown. PMID- 6285363 TI - Cytochrome c oxidase inhibition by anesthetics: thermodynamic analysis. AB - The thermodynamic parameters that characterize the inhibition of cytochrome c oxidase activity, in rat liver submitochondrial particles, by n-butanol, tetracaine, and dibucaine were obtained. Three equilibria were assumed in order to account for the data: for the interaction of inhibitor with the native state of the enzyme, for the interaction of inhibitor with the thermally (reversibly) denatured state, and for the change between the native and thermally denatured states. Inhibition results from interaction with both the native and denatured states but, because the interaction is stronger with the denatured than with the native state, the native/denatured equilibrium is shifted to the right by the anesthetics. The enthalpies of interaction are -2.3, -4.7, and 3.7 kcal/mol (1 cal = 4.18 J) for the native state and -10, -6, and -14 kcal/mol for the denatured state, for n-butanol, tetracaine, and dibucaine, respectively. These values are much smaller than the previous estimates obtained by using the assumption that anesthetics interact only with the thermally denatured state of enzymes (e.g., -81 kcal/mol for tetracaine inhibition of luciferase). Our results suggest that local anesthetics inhibit enzyme activity by causing a reversible perturbation of protein conformation. The magnitude of the perturbation is much smaller (in energetic terms) than that which accompanies thermal denaturation. PMID- 6285364 TI - Electrostatic potential molecular surfaces. AB - Color-coded computer graphics representations of the electrostatic potentials of trypsin, trypsin-inhibitor, prealbumin and its thyroxine complex, fragments of double-helical DNA, and a netropsin--DNA complex illustrate the electrostatic and topographic complementarity in macromolecule-ligand interactions. This approach is powerful in revealing intermolecular specificity and shows promise of having predictive value in drug design. PMID- 6285365 TI - Herpesvirus-specific RNA and protein in carcinoma of the uterine cervix. AB - Cloned probes of herpes simplex virus type 2 DNA were used in cytological hybridization experiments to detect herpesvirus RNA transcripts in the neoplastic cells of tumors of the uterine cervix. Virus-specific RNA was shown to represent transcription of limited regions of the genome, of which one is known to code for a DNA-binding protein that can be found by immunoperoxidase staining in the neoplastic cells of these tumors and has also been detected in cells transformed in vitro by this virus. PMID- 6285366 TI - Centruroides toxin, a selective blocker of surface Na+ channels in skeletal muscle: voltage-clamp analysis and biochemical characterization of the receptor. AB - This paper describes the effects of a toxin from the scorpion Centruroides suffusus suffusus on frog skeletal muscle. The main findings are the following, (i) Centruroides toxin (CssII) blocks the early phase of the inward sodium current in the muscle that arises from influx via Na+ channels in the surface membrane, but it does not affect the late phase of the inward current that represents flux through Na+ channels in the T-tubule membranes, (ii) CssII, in marked contrast to tetrodotoxin, does not affect contraction of the muscle, (iii) Measurements of the binding of 125I-labeled CssII to a partially purified membrane preparation from the muscle indicate that the Kd of the CssII--receptor complex is approximately 0.4 nM. The half-life for the dissociation of this complex is 3 min at 22 degrees C and 16 min at 2 degrees C. Binding of the radiolabeled toxin varies markedly with pH and becomes insignificant at pH greater than 8.5. Proteolytic digestion of the membrane preparation decreases its ability to bind CssII, suggesting that the receptor is a protein. (iv) The number of binding sites for a radiolabeled derivative of tetrodotoxin on the membrane preparation was similar to that for CssII. However, neither tetrodotoxin nor any of seven other neurotoxins and some local anesthetics that alter the functioning of the Na+ channel have any effect on the binding of CssII to the muscle membrane. These results therefore indicate that CssII belongs to a different class of neurotoxins that has a different receptor on the Na+ channel. PMID- 6285367 TI - Construction of plasmids for expression of Rous sarcoma virus transforming protein, p60src, in Escherichia coli. AB - We have constructed plasmids that direct the synthesis of the Rous sarcoma virus transforming gene (src) product (p60src) in Escherichia coli. A 203-base-pair lac promoter-operator DNA encoding the first eight amino acids of beta-galactosidase was ligated to the 5' end of the src gene from the Prague A strain of Rous sarcoma virus (PrA-RSV) which had been cloned in pBR325. Antiserum, from a tumor bearing rabbit, directed against pp60src was used to screen bacteria containing the recombinant plasmid for a protein of approximately 60,000 daltons, and several colonies producing a protein immunologically related to pp60src were detected. Partial proteolytic cleavage analysis revealed that the src-related protein produced in bacteria is structurally similar to pp60src immunoprecipitated from PrA-RSV-infected chicken cells. Partially purified src protein from E. coli can be phosphorylated in vitro by the catalytic subunit of cAMP-dependent protein kinase. Tryptic phosphopeptide analysis demonstrated that the catalytic subunit phosphorylated a serine-containing tryptic peptide in the bacterial src protein that comigrated with the phosphoserine-containing tryptic peptide of pp60src immunoprecipitated from 32P-labeled PrA-RSV-infected chicken cells. PMID- 6285369 TI - Phosphorus-containing inhibitors of angiotensin-converting enzyme. AB - Several phosphonamides, phosphoramides, and phosphates having the general structure R-Y-P(O)(OH)-X-CH(CH3)-CO-Pro have been synthesized and tested for inhibition of angiotensin-converting enzyme (dipeptidyl carboxypeptidase; peptidyl-dipeptide hydrolase, EC 3.4.15.1). Inhibition was found to depend on the nature of R, Y, and X such that the maximal effect was observed when X = NH, Y = CH2, and R = phi CH2 (50% inhibition at 7 nM). Substitution of CH2 or O at X and O at Y produced significantly less potent inhibitors. Groups shorter or longer than R = phi CH2 led to less active inhibitors, presumably due to nonoptimal interaction of the side chain with the S1 subsite. PMID- 6285368 TI - Surface receptors for pancreatic hormones in dog and rat hepatocytes: qualitative and quantitative differences in hormone-target cell interactions. AB - In order to evaluate potential differences in the kinetics of peptide hormone receptor interactions in hepatocytes of different species, we developed a simple procedure for the isolation of canine hepatocytes. Cells (obtained by collagenase perfusion of an extirpated dog liver lobe) were isolated with uniform high viability and yield. In addition, isolated dog hepatocytes tolerated incubation for at least 4 hr in defined medium with only a slight decrease in viability and with no change in the kinetics of [125I]iodoinsulin or [125I]iodoglucagon binding to cell-surface receptors. Comparisons of peptide hormone interactions with isolated dog and rat hepatocytes showed that (i) [125I]iodoglucagon associated with specific membrane receptors more rapidly than did [125I]iodoinsulin, for both rat and dog hepatocytes and at both 30 degrees C and 37 degrees C; (ii) the steady-state binding of [125I]iodoglucagon at 30 degrees C was greater than that of [125I]iodoinsulin in dog hepatocytes, but the reverse relationship held in rat hepatocytes; (iii) the rate of dissociation of [125I]iodoinsulin from hepatocytes of both species was enhanced by the presence of the unlabeled hormone, whereas the rate of dissociation of receptor-bound [125I]iodoglucagon was enhanced by the presence of unlabeled glucagon only in hepatocytes derived from the dog; and (iv) [125I]iodopancreatic polypeptide bound to neither rat nor dog hepatocytes, although the [125I]iodotyrosylated peptide bound to rat hepatocytes with an unusually high apparent dissociation constant. While confirming essential findings of pancreatic hormone binding to isolated hepatocytes, this comparison suggests that both qualitative and quantitative aspects of hormone-target cell interactions can show interspecies variability. PMID- 6285370 TI - Solubilization and purification of the alpha 1-adrenergic receptor using a novel affinity resin. AB - The highly selective alpha 1-adrenergic receptor antagonist prazosin was used to identify binding sites having alpha-adrenergic specificity in rat hepatic plasma membranes. Solubilization of the membrane-bound receptors was achieved by incubation with the nonionic detergent digitonin, and binding activity was assayed by using [3H]prazosin and a polyethylene glycol precipitation technique. Only 20-30% of the total receptor pool was released by the solubilization procedure. However, binding of [3H]prazosin was saturable [maximal value, 206 +/- 8 fmol/mg of protein (membrane) vs. 74 +/- 4 fmol/mg of protein (soluble)] and of high affinity [Kd, 0.6 +/- 0.2 nM (membrane) vs. 0.8 +/- 0.2 nM (soluble)]. To aid in purification of the receptors, an affinity resin was developed using an analog of prazosin, 2-(4-succinoylpiperazin-1-yl)-4-amino-6,7 dimethoxyquinazoline (CP 57,609; Kd 2.7 X 10(-7) M) immobilized via an amide linkage to agarose. The resulting resin demonstrated high affinity (Kd 3.2 X 10( 7) M) for the solubilized receptors, as determined by competitive inhibition assay. The degree of substitution to the resin was determined by a direct radioimmunoassay using antibodies against albumin-complexed CP 57,609 and found to be 0.1 to 0.2 mumol/ml of agarose. Affinity chromatography using the resin resulted in 513-fold purification in a single step. Moreover, the specificity of the purified binding sites was similar to that of membrane-bound receptors. This novel affinity resin should thus provide a powerful tool for isolating the receptor protein in quantities sufficient for detailed biochemical characterization. PMID- 6285371 TI - Beta-endorphin: opiate receptor binding activities of six naturally occurring beta-endorphin homologs studied by using tritiated human hormone and naloxone as primary ligands--effects of sodium ion. AB - Six naturally occurring homologs of beta-endorphin were tested for their potency in inhibiting the binding of [3H]naloxone and [3H]-beta h-endorphin either in the presence or in the absence of Na+. Sodium ion reduces inhibitory potency of these homologs against [3H]-beta h-endorphin and [3H]naloxone to about the same extent. The ratio of the IC50 obtained with Na+ to that obtained without Na+ is not highly correlated with analgesic potency. Very high (greater than 300) sodium ratios are observed with some homologs. A significant rank correlation was found between the ratio of the relative potency in displacing [3H]beta h-endorphin to the relative potency in analgesia and potency in inhibiting the binding of the pure antagonist naloxone. PMID- 6285372 TI - Separation of two thyrotropin binding components from porcine thyroid tissue by affinity chromatography: characterization of high and low affinity sites. AB - Two distinct thyrotropin (TSH) binding species have been separated from solubilized porcine thyroid membranes. Membranes was solubilized with 1% Triton X 100, and the supernatant was recovered by centrifugation at 105,000 X g. Scatchard analysis of thyrotropin binding to solubilized membranes (SM) yielded a nonlinear plot with Kd values for the high and low affinity components similar to those of intact membranes. Chromatography of the SM preparation on concanavalin A Sepharose 4B resulted in the retention of 10-20% of the binding activity. Upon elution of the column, a peak of binding material (5-7% of total activity) was eluted at 0.3 M alpha-methyl-D-mannoside. This concanavalin A (Con A) bound fraction exhibited a linear Scatchard plot with a Kd value similar to that of the high affinity component of the SM. The protein fraction that did not bind to Con A (Con A unbound) also exhibited a linear Scatchard plot, but with affinity similar to that of the low affinity component of SM. Discontinuous sucrose density gradient ultracentrifugation revealed the presence of two major binding peaks in the solubilized membrane preparation. The slowly sedimenting peak corresponded to that seen in the Con A bound fraction, whereas the rapidly sedimenting peak corresponded to that of the Con A unbound fraction. Sepharose 6B chromatography indicated that in the case of the Con A unbound fraction, a single peak of specific binding activity was eluted in the void volume, and in the case of the Con A bound fraction, one major peak with an approximate Stokes radius of 67 A and several other minor peaks were eluted. These results demonstrate the physical separation of two distinct TSH binding species from thyroid membranes and provide further support for the model of multiple classes of binding sites. PMID- 6285373 TI - Methylation of the viral genome in an in vitro model of herpes simplex virus latency. AB - An in vitro model of latency of herpes simplex virus type 1 (HSV-1) in a lymphoid cell line has been developed recently. CEM cells persistently infected with HSV-1 transiently ceased to produce virus for 24 days. This nonproductive state could either be reversed with phytohemagglutinin or maintained with concanavalin A. This system was used to study the relationship between DNA methylation and HSV-1 latency. DNA was probed for methylation by comparing the cleavage patterns generated by two pairs of restriction endonucleases (Sma I vs. Xma I and Hpa II vs. MspI); these enzymes show differential activity reflecting methylation of the recognition sequences. Viral DNA in the concanavalin A-treated cells (not producing virus) was found to be extensively methylated. By contrast, no methylated copies were detected in viral DNA from producer cells. About 800 days after the initial infection, the productive culture once again became nonproductive. Viral sequences in the latter cells were also methylated. Reconstitution experiments revealed 1-2 copies of viral DNA in cells from the latent stages and 40-80 copies in cells from productive stages. Most (if not all) of the viral genome is present in cells from various productive and latent stages. No differences in sequence arrangement were detected (although a terminal fragment of intracellular HSV-1 DNA appeared to be under-represented in latent cells). These results suggest a role for DNA methylation in the mechanism of HSV 1 latency in this system. PMID- 6285374 TI - Resistance to phosphatase of thiophosphorylated epidermal growth factor receptor in A431 membranes. AB - Epidermal growth factor (EGF) increases the phosphorylation of its receptor and other membrane proteins, and these proteins can be rapidly dephosphorylated by membrane-bound protein phosphatase [Carpenter, G., King L., Jr., & Cohen, S. (1979) J. Biol. Chem. 254, 4884]. We report that [35S]-adenosine 5'-[gamma thio]triphosphate is as effective as [gamma-32P]ATP as substrate for the EGF receptor-associated protein kinase in A431 membranes. Both the kinetics and the extent of the EGF-dependent thiophosphorylation at 0 degrees C are similar to those obtained with [gamma-32P]ATP, provided that ATP hydrolysis by the membrane preparation is inhibited by addition of adenosine 5'-[beta, gamma-imino] triphosphate. The thiophosphorylation reaction requires Mn2+ but differs from the phosphorylation reaction in the inability of Mg2+ to serve as cofactor. Both EGF dependent phosphorylated and thiophosphorylated membrane proteins yield the same two major bands of Mr 145,000-160,000 in autoradiograms of NaDodSO4/polyacrylamide gel electrophorograms. The rate of dephosphorylation of membrane proteins that have been thiophosphorylated in the presence of EGF is dramatically slower (factors of 1/20 to 1/40) than that of the phosphorylated proteins at both 0 degrees C and 32 degrees C. This increased metabolic stability of the thiophosphorylated proteins will be useful for investigation of the role of phosphorylation in the biological effects of EGF. PMID- 6285375 TI - Cloning of a yeast gene coding for arginine-specific carbamoyl-phosphate synthetase. AB - Several recombinant plasmids containing cpaII, the gene that encodes the large subunit of yeast arginine-specific carbamoyl-phosphate synthetase [carbamoyl phosphate synthetase (glutamine-hydrolyzing), carbon-dioxide: L-glutamine amido ligase (ADP-forming, carbamate-phosphorylating), EC 6.3.3.5], have been isolated. The plasmids were selected by transformation of a yeast strain with a mutation in the structural gene of the large subunit of carbamoyl-phosphate synthetase. By using a recombinant pool with inserts of yeast nuclear DNA of 5-20 kilobase pairs, we obtained 13 transformants. Of five transformants studied, three have been found to have stable plasmid inserts. These plasmids could be amplified in Escherichia coli and transferred back into the yeast carbamoyl-phosphate synthetase-deficient strains with concomitant complementation of the nuclear mutation. Plasmids pJL2/T1 and pJL2/T5 contain identical nuclear DNA inserts of 5.9 kilobase pairs. Although the insert of plasmid pJL2/T3 is also 5.9 kilobase pairs long, the sequence overlap with pJL2/T1 and pJL2/T5 is only 4.5 kilobase pairs long. The T3 insert has an orientation in the vector opposite to that of the T1 and T5 inserts. The recombinant plasmids with the yeast cpaII gene fail to cross-hybridize with a cloned fragment of E. coli DNA containing the carA and carB genes for the bacterial carbamoyl-phosphate synthetase. PMID- 6285376 TI - Use of monoclonal antibodies to subunits of human chorionic gonadotropin to examine the orientation of the hormone in its complex with receptor. AB - Monoclonal antibodies were prepared against the alpha and beta subunits of human chorionic gonadotropin (hCG). Although all were selected on the basis of their ability to bind the intact hormone, each also bound one of the two subunits but not both. Using a solid phase double antibody system to measure the relative binding to sites on the surface of hCG, we observed that four of the five antibodies bound to different sites on the molecule. This information was correlated with the ability of each antibody to inhibit the biological activity of hCG. Of the five antibodies tested for their ability to inhibit hCG-induced stimulation of rat testes steroidogenesis in vitro, two proved to be potent inhibitors, whereas the other three had almost no effect. This inhibition of steroidogenesis was highly correlated with the ability of the antibodies to inhibit hCG binding to testes homogenates. Thus, we have begun to derive a scheme that describes the relative binding positions of individual monoclonal antibodies and receptor on hCG. The purified monoclonal antibodies were iodinated and employed to evaluate which antigenic sites on hCG remained free in hCG-receptor complexes. The data indicated that portions of the beta subunit in hCG-receptor complexes were buried (i.e., failed to bind radiolabeled antibody), whereas other portions remained exposed (i.e., they bound radiolabeled antibody). Those antibodies that interacted with portions of hCG that became inaccessible in the receptor complex also blocked the biological actions of hCG, whereas those that interacted with exposed sites had little or no effect on activity. Although we did not find antibodies to the alpha subunit that would bind to the hormone receptor complex, we found that one of the two antibodies specific to alpha subunit epitopes blocked the actions of the hormone. Both antigenic determinants on the alpha subunit appeared to be lost after the hCG-receptor complex had formed. These studies suggest that each hCG subunit participates in the hormone receptor complex and that portions of the beta subunit project from the surface of the receptor. PMID- 6285377 TI - Introduction of rat growth hormone gene into mouse fibroblasts via a retroviral DNA vector: expression and regulation. AB - We have introduced the rat growth hormone gene into mouse fibroblasts via a retroviral DNA vector. The ability of the viral DNA to induce foci in the recipient cells was used as a dominant selection marker. Several copies of rat growth hormone DNA were integrated in the mouse cells. The transformed mouse cells expressed rat growth hormone-specific mRNA and secreted mature rat growth hormone. In rat cells, the expression of this gene is regulated by glucocorticoids. We demonstrate that hormone-dependent regulation transfers with the clone and thus appears to be an intrinsic property of the gene or its RNA products. PMID- 6285378 TI - Inheritance of DNA methylation in microinjected eggs of Xenopus laevis. AB - Plasmid DNA methylated at Hpa II sites was injected into eggs of Xenopus laevis. The methylated DNA replicates extrachromosomally, and the methylated state of the H pa II sites is inherited through replication. Unmethylated sites remain unmethylated in progeny molecules. To test whether replication is necessary for new methylation to occur, DNA methylated on one strand only was injected and unreplicated DNA was selected for analysis. Methylation was copied onto a previously unmethylated strand in the absence of replication but less efficiently than in its presence. These experiments show that the individual methylated site contains enough information for inheritance of the methylated state and that inheritance of methylation does not require integration of the DNA into the chromosome. PMID- 6285379 TI - Isolation and characterization of yeast mutants deficient in adenylate cyclase and cAMP-dependent protein kinase. AB - Mutants of Saccharomyces cerevisiae that require cAMP for growth have been isolated. Some of the mutants isolated were deficient in adenylate cyclase activity and mapped at a locus, cyr1, located near the centromere of chromosome X. Growth of cells carrying the cyr1 mutation was arrested at the G1 phase of the yeast cell cycle in the absence of cAMP. The cyr1 mutation was suppressed by a secondary mutation designated bcy1. The bcy1 mutation bypassed the need for cAMP for growth. The bcy1 mutants had extremely low levels of cAMP-binding protein and cAMP-dependent protein kinase but produced a high level of cAMP-independent protein kinase. The results indicate that cAMP is an essential factor for yeast cells to proceed through the cell cycle via the activation of protein kinase. PMID- 6285380 TI - Four small Drosophila heat shock proteins are related to each other and to mammalian alpha-crystallin. AB - The primary base sequence of the protein coding regions of the four small heat shock genes of Drosophila melanogaster present at cytological locus 67B has been determined. A single open reading frame large enough to encode a small heat shock protein is found for each gene. The molecular weights of the predicted proteins are in good agreement with experimentally determined values obtained from gel electrophoresis. The predicted amino acid sequences of the four small heat shock genes show striking homologies over approximately 50% of their lengths. This region of extensive homology extends from about amino acid 85 to amino acid 195 out of a total of approximately 200 amino acids. Comparison of the predicted sequence with the known sequences of other proteins revealed a remarkable similarity between this region of homology and the corresponding region of mammalian alpha-crystallin. The possible functional significance of this structural similarity is discussed. PMID- 6285381 TI - Human lung small-cell carcinoma contains bombesin. AB - The presence of immunoreactive bombesin in a human lung small-cell carcinoma grown in nude mice was established by several criteria: (i) Radioimmunoassay of tissue extracts for bombesin revealed approximately 6.5 pmol/g of tissue; (ii) bombesin was found in 12-14% of the tumor cells by immunohistochemical localization; (iii) gel filtration of small-cell carcinoma extract on Sephadex G 75 and Bio-Gel P-4 gave only a single peak of immunoreactivity, which occurred at the elution volume of bombesin; and (iv) reverse-phase HPLC of acid-solubilized extracts separated the immunoreactive material into three discrete peaks, one of which eluted with a retention time identical to that of synthetic bombesin. The presence of bombesin may represent the ectopic expression of this peptide in small-cell carcinoma, because immunoreactive bombesin was found in human fetal and neonatal lung but apparently not in adult lung tissue [Wharton, J., Polak, J. M., Bloom, S. R., Ghatei, M. A., Solcia, E., Brown, M. R. & Pearse, A. G. E. (1978) Nature (London) 273, 769-770]. The immunoreactive bombesin previously found in mammalian tissues is considerably larger than amphibian bombesin; these data substantiate the presence of a mammalian form of bombesin in a human tumor that may have a structure similar to that of the amphibian peptide. PMID- 6285383 TI - Triphenylmethylphosphonium is an ion channel ligand of the nicotinic acetylcholine receptor. AB - The lipophilic cation triphenylmethylphosphonium (Ph3MeP+), which is widely used as a sensor for membrane potential with cells, organelles, and membrane vesicles, is shown also to accumulate in membranes rich in nicotinic acetylcholine receptor in a voltage-independent way. Evidence is presented that Ph3MeP+ in this system is bound to a cation-binding site of the ion channel that is part of the acetylcholine receptor complex. Binding is stimulated by cholinergic effectors (Kd = 13 microM in the absence of carbamoylcholine; Kd = 1.5 microM in the presence of 10 microM carbamoylcholine), and this stimulation is blocked by alpha bungarotoxin. Ph3MeP+ blocks efflux of 22Na from receptor-rich microsacs and appears to compete with the channel ligand phencyclidine for a common binding site. In contrast to the binding of other proven channel ligands, Ph3MeP+-binding is not affected by desensitization. PMID- 6285382 TI - Construction of DNA recognition sites active in Haemophilus transformation. AB - Competent Haemophilus cells recognize and preferentially take up Haemophilus DNA during genetic transformation. This preferential uptake is correlated with the presence on incoming DNA of an 11-base-pair (bp) sequence, 5'-A-A-G-T-G-C-G-G-T-C A-3'. To prove that this sequence is the recognition site that identifies Haemophilus DNA to the competent cell, we have now constructed a series of plasmids, each of which contains the 11-bp sequence. Using two different assay systems we have tested the ability of fragments from these plasmids to compete with cloned Haemophilus DNA fragments that naturally contain the 11-bp sequence. We find that the addition of the 11-bp sequence to a DNA fragment is necessary and sufficient for preferential uptake of that fragment. However, plasmid DNAs containing this sequence may vary as much as 48-fold in uptake activity, and this variation correlates with the A+T-richness of the DNA flanking the 11-mer. PMID- 6285384 TI - Reactivation and microtubule sliding in rodent spermatozoa. PMID- 6285385 TI - Activation and reactivation of Ciona spermatozoa. PMID- 6285386 TI - Role of calmodulin in the flagellar axoneme: effect of phenothiazines on reactivated axonemes of Chlamydomonas. PMID- 6285387 TI - Palmar dermatoglyphics in Wilms' tumor. PMID- 6285388 TI - Prophylactic granulocyte transfusions. AB - Prophylactic granulocyte transfusions prevent infections in granulocytopenic marrow transplant recipients. In this setting the selection of a single "compatible" related donor for each recipient removes many of the immunologic problems inherent in utilizing multiple random volunteer donors. Graft-versus host disease is a real risk from granulocyte transfusions and can probably be prevented by irradiation prior to infusion. The problem of cytomegalovirus acquisition through granulocyte and other blood product donors remains to be solved. PMID- 6285389 TI - Selective stimulation of PGE1-synthesis by soterenol in the isolated rat vas deferens is mediated by alpha adrenoreceptors. AB - The effects of soterenol on prostaglandin (PG) release by the isolated rat vas deferens were studied. Soterenol increased the amount of PGE1 released into the medium without affecting that of PGE2 or PGF2 alpha. The effect of soterenol was inhibited by phentolamine but not by butoxamine. It is concluded that soterenol elicits the release of PGE1 by the isolated rat vas deferens via stimulation of alpha adrenoreceptors. PMID- 6285390 TI - Influences of tetrodotoxin, amiloride, phenytoin, aconitine, tetraethylammonium on ACTH-an isoproterenol-mediated lipolysis in isolated rat adipocytes. PMID- 6285391 TI - Dual effect of dihydroergotamine and dihydroergotoxin in isolated human femoral veins and arteries. AB - In human femoral vein strips, dihydroergotamine and dihydroergotoxin caused a slowly proceeding, long-lasting increase in tone. Dihydroergotamine had markedly higher affinity than noradrenaline, it possessed nearly the same intrinsic activity as noradrenaline. Dihydroergotoxin was less effective than dihydroergotamine. In isolated femoral artery strips, dihydroergotamine showed lower affinity and lower intrinsic activity in terms of noradrenaline. Dihydroergotoxin had negligible vasoconstrictor effects in arteries. In veins, dihydroergotoxin had a stronger alpha-adrenolytic effect than in arteries. In isolated arteries, dihydroergotoxin was equieffective to dihydroergotamine whose alpha-adrenoceptor blocking effect did not significantly differ in veins and arteries. Our results show that dihydroergotamine at very low concentrations exerted predominantly venoconstrictor effects and, therefore, it is more suited to increase the venous tone than dihydroergotoxin which at low concentrations showed alpha-adrenoceptor blocking activity. PMID- 6285392 TI - Effects of cannabinoids given orally and reduced appetite on the male rat reproductive system. AB - Chronic oral treatment of young adult male Fischer rats with delta 9 tetrahydrocannabinol (THC), 1, 5 and 25 mg/kg/day, or crude marihuana extract (CME), 3, 15 and 75 mg/kg/day, suppresses growth of accessory sex organs and body weight gain in a dose-related manner. Animals pair fed with the THC (25 mg/kg) group gained slightly more in body weight than the THC group, but their relative accessory sex organ weights were intermediate between THC and ad libitum-fed control group weights. These latter differences may be due to altered serum androgen levels since these levels 2-6 h after last treatment were 0.15, 0.77 and 3.33 ng/ml for THC, pair-fed and ad libitum-fed groups, respectively. 24 h after the last treatment all groups were within normal levels. Thus, chronic cannabinoid treatment suppresses accessory sex organ weights and serum androgen levels greater than the suppression caused by reduced food intake alone. PMID- 6285394 TI - Algebraic summation of the affective properties of a rewarding and an aversive stimulus in the rat. PMID- 6285393 TI - Neural plasticity: Part 2. Postnatal maturation and function-induced plasticity. AB - At birth, the central nervous system has completed most of its early stages of cell division, migration, and specialization. Much of the neural circuitry has been laid down. Neuroblasts are continuing to divide only in limited brain regions. Hence, at birth, most mammals have a nearly full complement of neurons. Nonetheless, the functional capabilities of the central nervous system of the newborn have little resemblance to those of the adult. Postnatal maturation must proceed in the proper sequence and at the proper rate if central nervous system deficits in the adult are to be avoided. Many of the prenatal maturational phenomena described in the previous paper continue well into the postnatal period. The purpose of this paper is to describe some postnatal maturational mechanisms and to show, by selected examples, the important role that function and experience play in central nervous system development. PMID- 6285395 TI - Kinetoplast DNA in the insect trypanosomes Crithidia luciliae and Crithidia fasciculata. I. Sequence evolution and transcription of the maxicircle. PMID- 6285396 TI - Kinetoplast DNA in the insect trypanosomes Crithidia luciliae and Crithidia fasciculata. II. Sequence evolution of the minicircles. PMID- 6285397 TI - Physical and genetic analysis of the Inc-W group plasmids R388, Sa, and R7K. PMID- 6285398 TI - A mutant of the plasmid R100.1 capable of producing autonomous circular forms of its resistance determinant. PMID- 6285399 TI - Selective trapping of circular double-stranded DNA molecules in solidifying agarose. PMID- 6285400 TI - Restriction endonuclease mapping of a Rhizobium leguminosarum Sym plasmid. PMID- 6285401 TI - Isolation and characterization of 2000-bp derivative of pBR322. PMID- 6285402 TI - A stable derivative of pBR322 conferring increased tetracycline resistance and increased sensitivity to fusaric acid. PMID- 6285403 TI - Inhibitors of cyclic AMP phosphodiesterase in medicinal plants. PMID- 6285404 TI - [Chemotherapy of bronchial carcinoma (author's transl)]. PMID- 6285406 TI - Action of cannabidiol on the anxiety and other effects produced by delta 9-THC in normal subjects. AB - The object of the experiment was to verify whether cannabidiol (CBD) reduces the anxiety provoked by delta 9-THC in normal volunteers, and whether this effect occurs by a general block of the action of delta 9-THC or by a specific anxiolytic effect. Appropriate measurements and scales were utilized and the eight volunteers received, the following treatments in a double-blind procedure: 0.5 mg/kg delta 9-THC, 1 mg/kg CBD, a mixture containing 0.5 mg/kg delta 9-THC and 1 mg/kg CBD and placebo and diazepam (10 mg) as controls. Each volunteer received the treatments in a different sequence. It was verified that CBD blocks the anxiety provoked by delta 9-THC, however this effect also extended to marihuana-like effects and to other subjective alterations induced by delta 9 THC. This antagonism does not appear to be caused by a general block of delta 9 THC effects, since no change was detected in the pulse-rate measurements. Several further effects were observed typical of CBD and of an opposite nature to those of delta 9-THC. These results suggest that the effects of CBD, as opposed to those of delta 9-THC, might be involved in the antagonism of effects between the two cannabinoids. PMID- 6285405 TI - The psychological experience of nursing mothers upon learning of a toxic substance in their breast milk. AB - This study explores the psychological reactions of a group of nursing mothers whose breast milk was inadvertently contaminated with a toxic fire-retardant chemical, polybrominated bihenyl. In particular, the extent to which the 97 mothers in the study denied the presence and possible effects of the chemical in their offspring is examined. Corresponding psychological defenses and reactions such as guilt or ambivalence in the nursing relationship and efforts at mastery of the situation are described with accompanying clinical illustrations. Insofar as the presence of a variety of toxic chemicals in the environment has been reported, the psychological reactions discussed here may have widespread application. PMID- 6285407 TI - Cellular lung dosimetry for inhaled radon decay products as a base for radiation induced lung cancer risk assessment. II. Microdosimetric calculations. AB - Lung dose calculations for inhaled radon decay products presented in part I have revealed that mean basal cell doses are significantly dependent on various personal and environmental factors. Whereas these macroscopic dosimetric methods have been applied with great success to radiation protection problems, the interpretation of radiobiological effects, such as lung cancer incidence, needs some refinement of these methods. Energy deposition at the microscopic level as the physical input quantity and radiation carcinogenesis as the biological endpoint are by nature stochastic processes. Therefore, a microdosimetric model was developed taking into consideration the randomness of physical and biological parameters involved, Part II of the paper presents results on specific energy distributions in lung cells, demonstrating that single event density distributions together with the number of cells receiving single hits represent more appropriate parameters than mean radiation doses. PMID- 6285408 TI - Cellular lung dosimetry for inhaled radon decay products as a base for radiation induced lung cancer risk assessment. I. Calculation of mean cellular doses. AB - Lung cancer induction is commonly regarded as the most important somatic risk arising from the inhalation of radon and its decay products. Relating carcinogenesis to radiation exposure needs a detailed knowledge of the cellular dose distribution in the human respiratory tract. Different dosimetric models have been developed for the determination of cellular doses, particularly for the basal cells of the bronchial epithelium which are considered as the critical cells for lung cancer induction. Part 1 of the paper describes the influence of various environmental as well as anatomical and physiological factors on the resulting dose. Significant inter- as well as intra-subject variabilities of structural components of the human lung, respiration characteristics and clearance mechanisms demonstrate the necessity of applying stochastic models in lung dosimetry. PMID- 6285409 TI - Biopsy measurement of DNA damage and repair in vivo: single-strand breaks, alkali labile bonds, and endonuclease-sensitive sites. PMID- 6285410 TI - Ultrasound in the first trimester of pregnancy. PMID- 6285411 TI - [Possibilities of rationalization in the field of lymphographic diagnosis. I. Reduction of the lymphographic exposure program for initial and control examination (author's transl)]. PMID- 6285412 TI - [Results of examinations on the improved resorption in fractionated oral cholangio-cholecystography with Falignost (author's transl)]. PMID- 6285413 TI - Angiographic classification of hepatic arterial collaterals. AB - Hepatic artery collaterals in 40 patients who had had hepatic artery occlusion following peripheral or central embolization, surgical ligation, intra-arterial chemotherapy, or intimal injury from catheterization were studied. The collaterals were classified as intrahepatic or extrahepatic collaterals. Intrahepatic arterial collaterals develop in the portal triads and subcapsular area between the lobes of the liver. Extrahepatic arterial collaterals develop in the ligaments that suspend the liver in the peritoneal cavity and through the structures that are closely attached to the liver. A simplified angiographic classification of hepatic arterial collaterals is presented. PMID- 6285414 TI - Dynamic, diagnostic, and pharmacological radionuclide studies of the esophagus in achalasia. AB - The esophagus was evaluated in 15 patients with achalasia by continuous gamma camera imaging following ingestion of a semi-solid meal labeled with 99mTc. The images were displayed and recorded on a simple computerized data processing/display system. Subsequent cine mode images of esophageal emptying demonstrated abnormalities of the body of the esophagus not reflected by the manometric examination. Computer-generated time-activity curves representing specific regions of interest were better than manometry in evaluating the results of myotomy, dilatation, and drug therapy. Isosorbide dinitrate significantly improved esophageal emptying. PMID- 6285415 TI - A noncomputerized count-based technique for the determination of resting left ventricular ejection fraction. AB - A noncomputerized, count-based technique for the determination of left ventricular ejection fraction (LVEF), which does not use geometric assumptions of left ventricular shape, was developed. The noncomputerized technique and computerized multigated ventriculography using both fixed and variable region-of interest (ROI) methods were performed on 16 patients. The LVEFs obtained with the noncomputerized technique correlated well with both the fixed ROI computerized technique (r = .87) and the variable ROI computerized technique (r = .86). It is concluded that when a computer is not available, the noncomputerized technique is a valid alternative for the determination of LVEF in resting patients in stable sinus rhythm. PMID- 6285416 TI - [An immunohistochemical attempt to demonstrate growth hormone or growth hormone binding sites in the human brain (author's transl)]. PMID- 6285417 TI - Poly(adenosine diphosphate ribose). PMID- 6285418 TI - Cyclic nucleotide control of protein kinases. PMID- 6285419 TI - tRNA-like structures in the genomes of RNA viruses. PMID- 6285420 TI - Dietary administration of eicosapentaenoic and linolenic acid increases arterial blood pressure and suppresses vascular prostacyclin synthesis in the rat. PMID- 6285421 TI - "Sand bar" of actinoid and neighboring elements in a chart of nuclides. AB - Inspection of a chart of nuclides reveals a "sand bar" of actinoid and neighboring elements when a suitable half-life level is chosen. A bay of alpha instability lying between the nuclides of neutron magic number 126 and the beta stability line in the actinoid group is a characteristic feature in this area. Calculation shows that fairly good agreement is obtained between the measured half-lives and those derived theoretically. PMID- 6285423 TI - Beam test results of small AVF-cyclotron for radioisotope production. PMID- 6285422 TI - Amogastrin for the 99mTc-pertechnetate imaging of the gastric mucosa. AB - Most of the gastric diseases seat in the mucosa, and the present investigation was undertaken in order to see if amogastrin, a synthetic tetrapeptide, was capable of enhancing gastric accretion of intravenous 99mTc pertechnetate in man as well as in the rabbit. Improvement of gastric scintigraphy was achievable with a pretreatment of the agent by increasing the gastric accretion and reducing the background radioactivity in the liver and small intestinum. PMID- 6285424 TI - [Incidence of the disease known as cystosarcoma phylloides in fibroadenomas of the breast]. PMID- 6285425 TI - [In vivo and in vitro effects of peptide extracts from Amanita virosa]. AB - Cyclic peptides from the deadly mushroom Amanita virosa has been separated by methanolic extraction and chromatography on Sephadex. Three groups of peptides have been obtained: virotoxins, amaninamide and phalloidin. Virotoxins has been separated in two fractions named virotoxins A and virotoxins B. We have studied the properties of these two fractions of F actin in vitro and on mice in vivo. Our results show that virotoxins A and B protect F actin in vitro against chaotropic ions, depolymerization by DNAse I or cytochalasin B and heat denaturation. Virotoxins A and B increase the rate of polymerization of F actin. Virotoxins A and B are toxic compounds which produce hemorrhagic necrosis of liver that have been observed in detail by electron microscopy. In general, our in vitro and in vivo results show that virotoxins exhibit the same effect as phalloidin on F actin. PMID- 6285427 TI - [Associated dermatomyositis and cholangiocarcinoma]. PMID- 6285426 TI - [Hormonal control of liver gluconeogenesis]. AB - The stimulation of gluconeogenesis by glucagon results from a concerted mechanism involving: 1) the stimulation of pyruvate transport and carboxylation in mitochondria; 2) the cyclic AMP dependent phosphorylation and inactivation of pyruvate kinase resulting in a re-routing of phosphoenolpyruvate towards glucose; 3) the inhibition of phosphofructokinase and the stimulation of fructose bisphosphatase resulting from the disappearance of fructose-2,6-bisphosphate. Catecholamines and vasopressin stimulate gluconeogenesis in starvation whereas in the fed state they promote glycogenolysis together with glycolysis. PMID- 6285428 TI - [Endocarditis caused by Serratia marcescens sterilized by cefotaxime and trimethoprim-sulfamethoxazole]. PMID- 6285429 TI - [Stimulation of gluconeogenic capacity in partially hepatectomized rats (author's transl)]. AB - Renal gluconeogenic capacity was enhanced to 150% 24 h after partial hepatectomy, remained increased at 48 h (144%) and returned to normal values at 72 h. Glucose production by renal cortical slices from hepatectomized rats was also enhanced 48 h after surgery when pyruvate, alpha-ketoglutarate and fructose were used as gluconeogenic precursors. The stimulation of renal gluconeogenic capacity seems to be due to the increase of phosphoenolpyruvate carboxykinase and glucose-6 phosphatase activities which behaved similarly to glucose production after hepatectomy. The renal metabolic response may be partially due to starvation in the first 24 h. Afterwards food intake became normalized and the acceleration of glucose production should be attributed to hepatectomy. Since there was no metabolic acidosis in our experimental conditions the involvement of glucocorticoids in the stimulation of renal phosphoenolpyruvate carboxykinase and glucose-6-phosphatase activities is suggested. PMID- 6285430 TI - [Small cell bronchial carcinoma. A general review (author's transl)]. PMID- 6285431 TI - Effect of exogenous adenosine 3':5'-cyclic monophosphate, parathyroid hormone and acetazolamide on electrolyte hormone and acetazolamide on electrolyte excretion by the isolated perfused rat kidney. AB - The effects of exogenous adenosine 3':5'-cyclic monophosphate (cyclic AMP), parathyroid hormone (PTH) and acetazolamide (Az) on renal calcium and phosphate excretion of the isolated perfused rat kidney were compared. Both PTH and Az evoked an early increase in urinary cyclic AMP excretion followed by a later more prolonged increase in phosphate excretion. All of the increased urinary cyclic AMP was derived from renal cells. Calcium excretion decreased with PTH but was unchanged with Az. Sodium and potassium excretion increased with Az but not PTH. Transitory urinary cyclic AMP excretion rates following bolus additions of exogenous cyclic AMP to perfusate were up to twentyfold greater than those evoked by PTH or Az but unassociated with changes in calcium, phosphate, sodium or potassium excretion. Sustained perfusate levels (above 0.5 microM) and excretion rates of cyclic AMP induced phosphaturia proportional to the perfusate cyclic AMP concentrations achieved up to 2.0 microM. Clearances of exogenous cyclic AMP exceeded inulin clearance at perfusate concentrations greater than 1.0 microM. Aminophylline evoked a small phosphaturia which increased further on addition of cyclic AMP to 5 microM. Glomerular filtration was not affected by any of the agents except Az. Since increased phosphate excretion could be evoked only at perfusate concentrations exceeding either plasma or intracellular concentrations observed in vivo, it is concluded that circulating cyclic AMP at levels in vivo is unlikely to mediate a significant fraction of the renal effects of PTH. PMID- 6285432 TI - Role of beta-adrenergic receptors in the antidepressant activity of alprazolam. AB - Chronic intravenous infusion of reserpine significantly increased beta-adrenergic receptors (beta-AR) in the cerebra cortex of rats. This increase in beta-AR is partially blocked by chronic treatment with alprazolam. However, similar effects were not observed with diazepam. These results indicate that beta-AR may be involved in the antidepressant activity of alprazolam. PMID- 6285433 TI - Immunosuppression in bovine trypanosomiasis: response of cattle infected with Trypanosoma congolense to foot-and-mouth disease vaccination and subsequent live virus challenge. AB - The primary and secondary antibody responses to foot-and-mouth disease virus vaccine were examined in cattle infected with Trypanosoma congolense and the response of some of these animals to live foot-and-mouth disease virus challenge was assessed. Infected groups of cattle had rather lower antibody responses than uninfected control cattle after primary vaccination but the antibody titres were not significantly depressed until after secondary vaccination. These levels remained depressed for the duration of the experiment, ie, 183 days. Trypanocidal therapy with diminazene aceturate of infected cattle at the time of vaccination did not significantly improve the antibody response to primary vaccination. Their subsequent response to live virus challenge was somewhat equivocal in that the number of animals protected was not significantly different in comparison to the untreated infected and uninfected controls. It was concluded that trypanosome infected cattle do not produce optimal responses to foot-and-mouth disease vaccination. Nevertheless, the antibody titres are generally above those considered adequate to confer 95 per cent protection against needle challenge. PMID- 6285434 TI - Immunoperoxidase study of Aujeszky's disease in pigs. AB - Viral antigen was detected by an immunoperoxidase technique in histological sections from pigs with Aujeszky's disease. The antigen was found mainly in association with focal necrosis in the cerebellum, tonsils, oral and nasal mucosa, salivary glands, lungs, liver, kidneys, pancreas, spleen and adrenal glands. Cells at the margin of the necrotic foci especially were strongly positive. Viral antigen was also demonstrated in the cerebral cortex and in the brain stem. Two types of intranuclear inclusion bodies were found to contain viral antigen and one type also contained viral nucleic acids. Inflammatory cells usually contained no viral antigen. The possible significance of some of these infected tissues in the excretion of the virus is discussed. PMID- 6285435 TI - Adrenocortical function and plasma biochemical values in dogs after subconjunctival treatment with methylprednisolone acetate. AB - The effects of subconjunctivally administered methylprednisolone acetate on adrenocortical function and blood composition were studied in five dogs. Two 10 mg doses of methylprednisolone acetate were injected at an interval of 21 days. Plasma cortisol concentrations (COR), ACTH-stimulated cortisol concentrations and other blood components were determined regularly. The normal plasma COR (5.7 +/- 2.62 ng/ml) was one of the lowest reported in the literature for the dog, possibly because of the specificity of the high performance liquid chromatographic assay and the experimental environment. Plasma COR was depressed only after the first administration of methylprednisolone acetate. ACTH stimulated COR was significantly depressed nine and 20 days after the first and second subconjunctival injection respectively. Otherwise blood composition was unchanged. PMID- 6285437 TI - Acute infection of a mediastinal cyst. PMID- 6285436 TI - Antibody response of pigs to foot-and-mouth disease oil emulsion vaccine: the antibody classes involved. AB - Groups of three pigs were vaccinated with water-in-oil emulsion vaccine and revaccinated either 21 and 148 or 106 days later. Sera were taken periodically for six months and fractionated into heavy and light elements on sucrose density gradients. The heavier fraction contained IgM and the lighter fraction IgG and IgA. Neutralising antibodies were first detectable eight days after initial vaccination (dpiv), rose to a peak between 14 and 21 dpiv and persisted at relatively high titres until the time of revaccination. Neutralising antibody at eight dpiv was attributed to IgM but by 10 to 14 dpiv both IgM and IgG were involved. Thirty-five days (and later) after a single vaccination all the neutralising activity in the sera was due to IgG. The revaccinations produced an increment in the whole serum neutralising titres and in each case both IgM and IgG class antibodies were involved. PMID- 6285438 TI - [Characterization of bovine herpesvirus type I Puente Alto Chile 1977 strain]. PMID- 6285441 TI - [Why does a limb "fall asleep'?]. PMID- 6285442 TI - [Physiological basis for acupuncture, electro-acupuncture and transcutaneous electrostimulation]. PMID- 6285439 TI - [Plasmacytosis with crystalline inclusions. Apropos of 5 cases]. PMID- 6285440 TI - [CCI4 - induced liver cirrhosis, in the rar: effects of colchicine in its induction and recovery]. PMID- 6285443 TI - [Irritable colon]. PMID- 6285444 TI - [Immunological and therapeutic aspects of myasthenia gravis (author's transl)]. AB - In the treatment of Myasthenia Gravis (M.G.) new therapeutic techniques have been gradually added to the usual anticholinesterase drugs. Steroids, immunosuppressive drugs, plasma exchange have been employed whereas thymectomy is still the most useful surgical treatment. On the basis of both personal experience and data from the literature the AA. reviewed the therapeutic techniques. Thymectomy is practically indicated for all patients particularly young ones. However there are some reserves in the presence of thymomas as sometimes deterioration may occur. Steroids are mostly employed in cases resistant to other therapies. Immunosuppressive drugs are usually associated with plasma-exchange. The latter has been recently preferred because of the absence of side effects; however its real therapeutic value has not yet been fully assessed. PMID- 6285445 TI - [Controlled study of nifedipine in the treatment of Raynaud's phenomenon]. AB - In each of the 16 patients included in our first study [6 idiopathic Raynaud's phenomenon (I), 4 associated with systemic lupus erythematosus (SLE) and 6 with progressive systemic sclerosis (PSS)] digital vasospasm could be reproduced by immersion of both hands in cold water (4 degree C). Each patient received in a double-blind manner and random order on two consecutive days, the calcium-channel blocking agent nifedipine (20 mg) and placebo. Nifedipine protection against vasospasm provoked by cold water (4 degrees C) was considered good or excellent in 14 of the 16 patients (p less than 0.001 versus placebo). In the second study, 30 patients [12 I, 10 PSS, 5 SLE and 3 rheumatoid arthritis (RA)] received in a double blind manner and random order, on two consecutive weeks, nifedipine (20 mg 3 time daily) and placebo. The improvement with nifedipine (in percentage of the decrease of the number of vasospastic attacks) was 90.95 in the 1 group, 78.63 SLE and RA and 64.02 in PSS (p less than 0.01). An open study during 3 months has confirmed the effectiveness of nifedipine (10 mg 3 times daily). The improvement was 88.92 in the 1 group, 76.33 in SLE and RA and 59.16 in PSS, 7 out of 30 patients stopped the treatment because of side effects (headache, flush, nausea, oedema of the ankles). Thus nifedipine appears to be extremely useful in the treatment of Raynaud's phenomenon. PMID- 6285446 TI - [Mandibular involvement in cancer. Isotope and radiologic evaluation]. PMID- 6285447 TI - Intrathecal immune responses in mumps meningitis patients. AB - The in vitro 3H-thymidine incorporation of peripheral blood (PB) and cerebrospinal fluid (CSF) lymphocytes from 11 mumps meningitis patients was studied after stimulation with non-specific mitogens and microbial antigens. Although corresponding viral antibodies were always found in the CSF of seropositive patients by a sensitive radioimmunoassay, their intrathecal synthesis was directed mainly against mumps virus. Most of the patients had PB lymphocytes that reacted on stimulation with phytohaemagglutinin (PHA) or pokeweed mitogen (PWM), but a smaller number of them had reactive CSF cells. Only four patients showed stronger responses to PHA in the CSF than in PB. Most patients had mumps-reactive lymphocytes in PB but only two of them in the CSF. In contrast, these patients more often had increased CSF cell reactivity when tested with purified protein derivative, measles, and herpes simplex virus antigens. The results could not be explained by a damaged blood-brain barrier alone but may reflect the immunological status of the brain compartment in these patients. PMID- 6285448 TI - Detection of antigen-specific cellular immune response by the in vitro production of T-cell growth factor. AB - The amount of T-cell growth factor (TCGF) produced in vitro in antigen-stimulated lymphocyte cultures was measured by induction of growth in cell lines dependent on the factor. The method distinguished eight herpes-simplex-seronegative subjects from ten seropositive ones, and these results correlated with those of the lymphocyte blast transformation (LBT) test. The LBT responses and TCGF amounts produced in cultures with purified tuberculin (PPD) were also in good correlation (r = 0.833, P less than 0.001). Considerable levels of the factor were already present 12 h after the beginning of the culture. The method offers rapid quantification of cell-mediated immunity to specific antigens. PMID- 6285449 TI - Serological diagnosis of pertussis: IgM, IgA and IgG antibodies against Bordetella pertussis measured by enzyme-linked immunosorbent assay (ELISA). AB - Igm, IgA and IgG antibodies against Bordetella pertussis were measured by enzyme linked immunosorbent assay (ELISA) with an ultrasonicate of formalin-killed bacteria (a mixture of strains 1, 2 and 1, 2, 3) as antigen and disposable polystyrene 9-cuvette blocks as the solid phase. The specificity properties of the assay were assessed by an inhibition technique. Of the microbes tested, only B. parapertussis was able to cause a significant inhibition. In addition, IgM and IgA antibodies against B. pertussis were only found in some sporadic cases of respiratory infections caused by other microbes. Sera, nasal swabs and cough plates were received from 198 patients with suspected whooping-cough. ELISA determinations were mostly made from only one serum sample of each patient. Paired sera were studied only from the culture-positive infants under 3 months of age. The number of positive cultures was highest in group under 3 months of age (41%), where the frequency of positive ELISA was lowest (20%). The use of paired sera strikingly increased the number of ELISA-positive individuals in this youngest patient group. In later life, the relationship between these tests changed: isolation was positive in only about 10% of the patients, whereas 29-64% yielded positive titres in ELISA. This study shows that pertussis ELISA is a valuable aid in the rapid diagnosis of pertussis, particularly of the atypical forms of the disease which mostly are culture-negative. PMID- 6285451 TI - Studies of cytomegalovirus infection in renal allograft recipients. AB - A prospective study of cytomegalovirus (CMV) infections has been carried out in 28 renal graft recipients. The protocol called for frequent blood and urine sampling during the first year after transplantation, but death or graft loss caused earlier termination in nearly half the patients. In this material 5/7 (71%) susceptible patients developed primary infections and 20/21 experienced a secondary infection (95%). Viruria was detected in 79% and viremia in 43%. The type of blood cell responsible for the viremic phase was studied by separating the blood cells on a density gradient. The polymorphonuclear cell fraction was the most common source of virus but virus could also be recovered from the mononuclear cell fraction. As some samples that were freeze-thawed repeatedly never yielded virus, it would appear that viable cells are needed for virus isolation. In both primary and secondary infections isolation of CMV from blood cells often preceded the isolation of CMV from urine. Among variables tested for a possible relationship to the occurrence of CMV viremia the only one to display such an association was the time at which rejection episodes occurred. In 19/28 such episodes recorded in 19 patients there was a temporal relationship to viremia (p less than 0.03). Seven of the patients experienced clinical symptoms suggestive of CMV infection as fever, cough, myalgia, arthralgia, chest pain and pneumonia. Laboratory signs included elevated amino acid transferase levels, leukopenia and thrombocytopenia and a specific anti-CMV antibody response. PMID- 6285450 TI - A fixed combination of pivmecillinam and pivampicillin in complicated urinary tract infections. A double-blind comparison with pivmecillinam. AB - 23 urological patients, primarily with underlying disease of the urinary tract, received a 10-day treatment with either pivmecillinam/pivampicillin or pivmecillinam alone. Combination therapy eradicated urinary pathogens in the urine at a 48 h control in 11/11 patients treated with the combination compared with 6/12 patients receiving pivmecillinam alone (p less than 0.05). Two of the treatment failures on pivmecillinam alone were caused by Enterobacteriaceae that were in vitro sensitive to mecillinam. A repeated course with the combination was successful in both of these patients. Apart from a rash seen in one patient in the combination group, no side effects were recorded. PMID- 6285453 TI - A prospective study of oropharyngeal excretion of Epstein-Barr virus in renal homograft recipients. AB - This prospective study was undertaken to examine the incidence of oropharyngeal excretions of Epstein-Barr virus (EBV) in the period after transplantation in human renal allograft recipients. Throat gargles from 23 recipients were tested for EBV. Five patients were already EBV excretors when the first specimens were tested. Of the remaining 18 patients 15 converted to positive within the first 7 months following transplantation. Three patients never excreted EBV in their throat gargles. This prospective study showed that 15/18 (83%) renal homograft recipients acquired active EBV infection (as indicated by excretion of EBV) after transplantation; 10/15 (66%) acquired active infection between 3-7 months after transplantation. We conclude that renal homograft recipients are highly susceptible to active EBV infection especially after the second month of transplantation. PMID- 6285452 TI - Studies of cytomegalovirus infection in renal allograft recipients. II. Serological response to various viral antigens. AB - In a prospective study the antibody response to various cytomegalovirus (CMV) antigens was examined in 28 renal allograft recipients. Both primary and secondary infections were investigated. Antibodies against immediate early (IEA) and early antigens (EA) were studied by anti-complement immunofluorescence; IgM and IgG antibodies to nuclear late antigens were differentiated by enzyme-linked immunosorbent assay (ELISA). The results of the tests were compared with each other and with those of the complement fixation (CF) test. 5/7 susceptible patients (71%) contracted primary infections. Both IgM and IgG antibodies developed and antibodies to IEA and EA appeared somewhat later. The antibodies to IEA and EA remained detectable throughout the observation period. Secondary infections developed in 20/21 (95%) patients. All initially had CMV antibody levels in ELISA and CF. Rising CMV titers of IgG antibodies were taken as a measure of secondary infection. IgM antibodies developed in only 10/20 (50%) patients. The highest titers of CMV IgM antibody levels were lower in secondary than in primary infections. Antibodies to IEA and EA were present prior to transplantation in some patients, but did not develop in all with secondary infections. The antibody titers were lower just after than before the transplantation in some patients. but subsequently increased again. It thus seems as if the humoral immune response to these CMV antigens differs in primary and secondary infections. PMID- 6285454 TI - Metronidazole in the treatment of anginose infectious mononucleosis. PMID- 6285455 TI - [Recurring tumors of the parotid gland with special reference to pleomorphic adenoma]. AB - 161 patients with parotid tumors seen between 1974 and 1980 are analyzed. A recurrence rate of 20% was found for both benign and malignant tumors. 76% of the recurrent tumors were judged to have been inadequately excised at the first operation. Pleomorphic adenomas had a recurrence rate of 25%. More than half these tumors exhibited multinodular patterns of recurrence. Therefore, a recurrent pleomorphic adenoma should not be reoperated upon for at least one year. PMID- 6285456 TI - [Detection of IBR/IPV antibodies in milk]. PMID- 6285457 TI - [Granulosa and theca cell tumors in cattle. (Observations in 13 cases)]. PMID- 6285458 TI - [Mechanisms of nerve impulses (author's transl)]. PMID- 6285460 TI - [Microwave metabostat and its application (author's transl)]. PMID- 6285459 TI - [Relationship of calcium ion with opiate and endorphins (author's transl)]. PMID- 6285461 TI - Calmodulin. PMID- 6285462 TI - Delineation of aspergilloma and granuloma with radiogallium and radiostrontium. PMID- 6285463 TI - E-rosette formation with SRBC and changes in cAMP level of hog lymphocytes--a study of certain affecting factors. AB - A further study of cAMP change in porcine lymphocytes after E-rosette formation is presented in the present report using peripheral blood lymphocytes of pig mixed with different concentrations of SRBC membrane as well as SRBC treated with different enzymes for modification of the rosette formation. It has been found that cAMP enhancement in lymphocyte follows the increase in the concentration of SRBC membrane used, and that a similar relation appeared between modification of cAMP and enzyme-treated SRBC in the rosette formation. It has also been found that after rosetting with SRBC, cAMP level of lymphocytes began to rise within 1 min, reached its peak at 10 min and then returned to control level in about 8 hr. PMID- 6285465 TI - Transfer of radiation-induced spins from deoxyribonucleic and thymidylic acids to propyl gallate. AB - When calf thymus deoxyribonucleic acid (DNA), propyl gallate (PG) and their five molecular mixtures (with PG content of 1, 2, 5, 10 and 20%) are irradiated with gamma-rays in dry state in vacuum at 296 degrees K, the ESR spectra of all molecular mixtures differ strikingly from those of DNA, but bear a close resemblance to those of PG. The spin yield in the PG contained in these mixtures is two to three orders of magnitude higher than that in the case of PG irradiated separately. Furthermore, on the basis of the relative saturation characteristics of ESR spectra, these molecular mixtures behave more like PG than like DNA. It may be inferred that the radiation-induced spins could be transferred from DNA to PG. With r representing the molar ratio of nucleotides to PG, we have found a good linear correlation between the transfer ratio (TR) and square root of r. One PG molecule could protect at least 68 nucleotides in the duplex DNA chain, and thereby the minimal range of spin transfer is estimated at 115 A. Results obtained from irradiation at 77 degrees K show that PG exerts no protective effect on DNA, so DNA sustains an irreversible damage. It is thought that the spin transfer from DNA to PG is exclusively due to a hydrogen transfer mechanism. We have also demonstrated the transfer of radiation-induced spins from both thermally denatured DNA and TMP to PG. The former process can be ascribed primarily to the hydrogen transfer mechanism, whereas the latter, as in the case of native DNA, exclusively to this mechanism. PMID- 6285464 TI - Constructions of gene libraries of Rana nigromaculata and Mus musculus with cosmid pHC79. AB - Gene libraries of Rana nigromaculata and Mus musculus (C57J6) have been constructed by the cosmid system developed by Collins using BamHl and Pstl as cloning restriction enzymes. A simplified formula, which is a modification of Clarke and Carbon derived from approximation, was used for calculating the number of colonies or recombinants necessary for the construction of a gene library. Pro+ and leu+ complementary to the recipient cells (HB101) with the genetic constitution of pro- and leu- have been selected from the BamH1 gene libraries of both Rana nigromaculata and the mouse in the present work. PMID- 6285466 TI - Studies on the incorporation of membrane proteins into liposomes: --effect of "boundary lipids" on reconstitution of pig heart mitochondrial cytochrome oxidase into liposomes. AB - The reactivation of delipidated cytochrome oxidase depends on the reformation of "annular lipids", which is tightly bounded to the enzyme molecule. In the restoration of oxidase activity, the efficiencies of phospholipids with different polar head groups decrease in this order: PS greater than DPG greater than PI greater than PA greater than PG greater than PC, PE and in the case of phosphatidylcholines with different acyl chain the order is DOPC greater than LPC greater than PC greater than DPPC, DSPC. Therefore both the polar head group and the acyl chain of phospholipids must be considered in the reactivation process. The existence and the specificity of "annular lipids" obviously influence the incorporation of cytochrome oxidase into liposomes. When acidic phospholipids are used as "annular lipids", the effectiveness of reconstitution decreases in this order: PI greater than PS greater than DPG greater than PA, PG. Divalent metallic cations would facilitate the cytochrome oxidase reconstitution, but their effects depend on the composition of "annular lipids". Using dialysis method Ca2+ and Mg2+ could facilitate the incorporation into liposomes of the enzymes having PS or DOPC as their "boundary lipids". A comparison between the effects of different metallic cations on incorporation of cytochrome oxidase also shows that, with PI as "annular lipids", the effectiveness of different cations on incorporation by incubation method decreases in this order: Ca2+ greater than Mg2+ greater than Mn2+, Sr2+ greater than La3+. Apparently, the effect of metallic cations on incorporation cannot be interpreted by considering only the neutralization of the negative charged groups on membrane protein and lipids. PMID- 6285467 TI - Sixty-three year old Caucasian female with cough and persistent pulmonary infiltrate. PMID- 6285468 TI - Opposite effects of inhaled cadmium microparticles on mouse susceptibility to an airborne bacterial and an airborne viral infection. AB - An experimental study on 489 mice is reported. The test animals were submitted to a single 15-mn exposure to atmosphere containing about 10 mg of cadmium microparticles (CdO) per m3 of air and the controls to an equivalent amount of aluminium microparticles (Al2o3). At the 48th hour after exposures, the test and control mice were submitted to a bacterial (Pasteurella multocida) or to a viral (Orthomyxovirus influenzae A) challenge, via the respiratory route. The exposure to cadmium significantly increased the death-rate of mice submitted to the bacterial challenge, but it significantly decreased the death-rate following the viral challenge. PMID- 6285469 TI - Mucus secretion-stimulating activity in human lymphoblastoid cells. AB - Two fractions isolated from cultured lymphoblastoid cells stimulated mucus secretion from the urn cell complex of the marine invertebrate Sipunculus nudus. The activity detected in the nuclear fraction was trypsin-sensitive, and it increased in response to specific nucleotides. PMID- 6285470 TI - Prolactin and growth hormone release by morphine in the rat: different receptor mechanisms. AB - Concentrations of prolactin and growth hormone in the serum of rats were significantly increased by morphine. Dose response studies demonstrated that maximum prolactin release required lower doses of morphine than those needed for the maximum growth hormone response. Selective blockade of mu 1 (high affinity) opiate receptor with the irreversible antagonist naloxazone reduced morphine induced peak concentrations of prolactin by 80 percent while increasing peak growth hormone levels by 250 percent. These results suggest different receptor mechanisms for the opiate modulation of the two hormones. The mu 1 (high affinity) receptor sites appear to mediate the morphine-induced release of prolactin but not growth hormone. PMID- 6285471 TI - Cellular transforming genes. AB - Cellular genes potentially capable of inducing oncogenic transformation have been identified by homology to the transforming genes of retroviruses and by the biological activity of cellular DNA's in transfection assays. DNA's of various tumors induce transformation with high efficiencies, indicating that oncogenesis can involve dominant genetic alterations resulting in activation of cellular transforming genes. The identification and characterization of cellular transforming genes and their possible involvement in naturally occurring cancers, is discussed. PMID- 6285472 TI - Phosphorylation of myosin light chains in mouse fast-twitch muscle associated with reduced actomyosin turnover rate. AB - Phosphorylation of the 18,000-dalton light chains of the fast-twitch myosin in mouse extensor digitorum longus muscles was correlated with reduction in the rate of the actomyosin adenosinetriphosphatase in vivo, but neither of these changes occurred in the soleus muscle. These results suggest that actomyosin interactions can be down-regulated by a reversible covalent modification of myosin light chains, that a mechanism for thick-filament regulation occurs in vertebrate skeletal muscle, and that the expression of this regulation may be limited to a specific fiber type. PMID- 6285473 TI - Endotoxin-stimulated opioid peptide secretion: two secretory pools and feedback control in vivo. AB - Small doses of endotoxin evoked a dramatic biphasic response of opioid peptide secretion into blood in sheep. The first phase began within minutes and coincided with a brief hypertensive response to endotoxin well before the appearance of fever or hypotension. The ratio of beta-endorphin to beta-lipotropin fell abruptly at the onset of the second phase of release, suggesting early depletion of a pool rich in beta-endorphin and subsequent emergence of a pool rich in unprocessed precursor. The concentration of cerebrospinal fluid opioids increased tenfold during the second phase. Naloxone administration augmented endotoxin induced opioid secretion in both early and late phases, suggesting a short-loop feedback regulation of stress-induced endorphin secretion. PMID- 6285475 TI - [Hepatic tuberculosis: anatomical and clinical findings. A review of forty-six cases (author's transl)]. AB - Over eight years the authors recorded 46 cases of hepatic tuberculosis. Most cases occurred in patients from black Africa. Forty-five patients had at least one extrahepatic tuberculous localization. Moderate cholestasis, without jaundice, found in 75% of patients, was the main biologic disorder. Upon histologic examination, five different types of granulomas were found. Each one has a specific predictive value concerning the outcome of the tuberculous infection. PMID- 6285474 TI - Significance of tissue myo-inositol concentrations in metabolic regulation in nerve. AB - Approximately 25 percent of resting energy utilization in isolated nerve endoneurium is inhibited by medium containing defatted albumin and selectively restored by arachidonic acid but is unaffected by indomethacin or nordihydroguaiaretic acid. The same component of energy utilization is inhibited by small decreases in endoneurial myo-inositol, which decrease incorporation of carbon-14-labeled arachidonic acid into phosphatidylinositol. The fraction of the resting oxygen uptake inhibited by ouabain is decreased 40 to 50 percent by a reduced tissue myo-inositol concentration or by defatted albumin. Metabolic regulation by rapid, basal phosphatidylinositol turnover is dependent on the maintenance of normal tissue myoinositol concentrations. PMID- 6285476 TI - [Colic carcinoma and basal cell epithelioma eleven years after Kaposi sarcoma with splenic and cutaneous involvement (author's transl)]. AB - A case of Kaposi sarcoma (KS) with cutaneous and splenic involvement is reported. Carcinoma of the left colon and basal cell epithelioma arose eleven years after KS. Genetic, viral, and immunologic factors which may promote initiation and development of Kaposi sarcoma are reviewed. The responsibility of immunodeficiency, whether resulting from therapy or from other causes, in carcinogenesis is discussed. Prolonged survival may be seen after KS. Another primary malignant disease (lymphoma or solid tumor) may arise. The authors suggest that the risk of immunologic disorders, occurring spontaneously or induced by therapy, should be considered specifically for each patient with KS. PMID- 6285477 TI - [Primary reticulum-cell sarcoma (Parker-Jackson sarcoma) of the rib. Report of a case with a five-year survival after surgery, radiation therapy and chemotherapy (author's transl)]. AB - A case of primary reticulum-cell sarcoma of the rib is reported. This rare tumor arose in an unusual site and was revealed by an exceptional clinical picture. Extensive hemothorax was the initial manifestation. Follow-up exceeds five years. Prognosis is not as poor for reticulum-cell sarcoma as for Ewing sarcoma. After radical surgery, radiation therapy and chemotherapy, the five-year total remission rate approximates 50%. PMID- 6285479 TI - [Osteo-onychodysplasia (nail-patella syndrome): a case-report (author's transl)]. AB - A case of osteo-onychodysplasia (nail-patella syndrome) is reported. Features of this inherited condition include hypoplastic nails, hypoplastic to absent patellae, subluxation of the patellae, elbow dysplasia, and spurs in the posterior ileum (this last feature was absent in our patient). The nail hypoplasia is the most suggestive abnormality. Patients should be investigated for renal involvement which is a poor prognosis factor. In patients without renal involvement, prognosis is good. PMID- 6285478 TI - [Stenosis of the right pulmonary artery secondary to carcinoma of the left lung. Effect of radiation therapy (author's transl)]. AB - A case of undifferenciated carcinoma of the left lung with stenosis of the right pulmonary artery is reported. The characteristic clinical features of the systolic ejection murmur in pulmonary artery stenosis are recalled as well as the data from phonocardiographic, angiographic, and hemodynamic investigations. After radiation therapy, the grade of the systolic murmur decreased frankly although transiently. Noticeable decrease of the stenosis was substantiated by a second angiography. This improvement was still apparent 16 months later during chemotherapy. PMID- 6285481 TI - [Two cases of pentastomiasis observed in Paris (author's transl)]. AB - The observations of two black African patients seen among 940 hospitalized patients are reported. Intraabdominal calcified larvas, probably of Porocephalus (Armillifer) armillatus were found in both patients. The cycle of this Pentostomida is recalled. Attention is drawn to the role of snake slime in accidental contamination of human beings. In man, this parasitic infection is almost always asymptomatic and detected fortuitously on roentgenograms, or during surgery or necropsy. PMID- 6285480 TI - [Toxopachyosteosis of the tibial and fibular diaphyses (Weismann-Netter and Stuhl syndrome) (author's transl)]. AB - Two cases of toxopachyosteosis involving the tibial and fibular diaphyses are reported. This rare condition, which is usually well tolerated, was first described by Weismann-Netter and Stuhl in 1954. This dysplasia affects both tibias and fibulas and usually results in symmetrical deformations. Characteristic features are anterior bowing of affected bones, with posterior thickening of the cortices. There are few clinical manifestations; small stature, often with delayed onset of walking. Biological investigations are negative. Roentgenograms often show embedding of the last lumbar vertebra with horizontalization of the sacrum. Scoliosis is a common finding. PMID- 6285482 TI - [Malignant histiocytosis. A histological and cyto-pathological study of twenty five cases in children (author's transl)]. PMID- 6285483 TI - [Melorheostosis, osteopoikilosis and linear scleroderma (author's transl)]. PMID- 6285484 TI - [Median nerve compression syndrome at the elbow (author's transl)]. AB - Twelve cases of median nerve compression at the elbow are presented. More than half of them showed only an anterior interosseous nerve syndrome (Kiloh-Nevin). Nevertheless, surgical exploration was performed from the brachial tunnel to the superficialis arch. When the neurolysis occurred reasonably early, uniform good results were obtained. PMID- 6285485 TI - [Clinical and radiological findings in 203 cases of delayed resorption of pulmonary fluid (author's transl)]. AB - Among 1 186 neonates admitted consecutively to the Post-Royal intensive care Unit, 203 had respiratory distress, a chest roentgenogram typical of delayed resorption of lung fluid, and no other findings. Mean gestational age (GA) was 33 weeks and mean birth-weight (BW) was 1 948 g. Hood oxygen alone was used in 129 cases and intubation-ventilation-PEEP in 74 cases (46 of which were intubated before admission). The occurrence of radiological features was similar in ventilated and non-ventilated neonates : ground-glass pattern (70-80% of the cases), increased vascular markings (70-76%) enlarged right fissure (45%), thin lateral pleural density (24%) and normal cardio-thoracic ratio. The analysis of other prognostic factors showed that ventilated neonates had a significantly lower birth weight (1 689 g versus 2 097 g) and a significantly younger gestational age (31.9 versus 33.6). The study of DAa02 in non-ventilated neonates demonstrated wide individual variations, and the possibility of an early normal DAa02 but also of a late abnormal DAa02, with the associated risks of early hyperoxia or prolonged oxygen needs. In ventilated neonates, the mean duration of intubation was 4 days 13 hours; this duration decreased as the birth-weight rose. In view of the major role of BW and GA, the Port-Royal team advocates the direct admission to a neonatal intensive care unit of neonates with delayed resorption of lung fluid and a BW under 2 000 g. In this way, Pa02 can be closely monitored and artificial ventilation is readily available. PMID- 6285486 TI - [Transbronchial biopsy of the lung : value for diagnosis of sarcoidosis. A study in 53 patients (author's transl)]. AB - Data yielded by bronchial and transbronchial biopsy specimens removed during fiberoptic bronchoscopy in 53 patients with sarcoidosis (24 stage I, 27 stage II, and 2 stage III) was analyzed. Bronchial biopsies (3 to 5 specimens) were positive in 49% of patients (41,6% stage I, 55,5% stage II, 1/2 stage III). Transbronchial biopsies removing bronchiolar and alveolar specimens yielded 88,6% positive results (91.6% stage I, 85,1% stage II, 2/2 stage III). Lymphocyte counts bronchioalveolar washings exceeded normal values by 15% in all 42 patients studied. We conclude : 1) that transbronchial lung biopsies yield significantly more information than proximal bronchial biopsies ; 2) that results of bronchioloalveolar washings reflect granulomatous immunologic changes in the lung and do not allow unequivocal diagnosis. PMID- 6285488 TI - [Semiology of spinal meningiomas and neurinomas (author's transl)]. AB - The last 60 patient who underwent surgery for neurinoma or meningioma of the spine provide the basis for this report. One-fourth of all cases of spinal cord compression are due to meningioma or neurinoma. Attention is drawn to the characteristic clinical features of these tumors, particularly their time-course, since diagnosis if often delayed. The sooner diagnosis is established, the better postoperative recovery will be, especially in elderly patients (60% of cases). Results of operative treatment are presented. PMID- 6285487 TI - [Neurosyphilitic arteritis. Clinical, paraclinical and therapeutic data. A review of six cases (author's transl)]. AB - Among forty-six patients with neurosyphilis, seen over a ten-year period, six had features highly suggestive of syphilitic cerebral vascularitis. The carotid distribution was involved in five patients and the posterior cerebral artery in one. As exemplified by these six observations, syphilitic arteritis should be considered when an ischemic stroke occurs with no apparent etiology. The fluorescent treponemal antibody adsorbed test (FTA-ABS) in the cerebrospinal fluid is the most reliable and the most specific laboratory test for the diagnosis of neurosyphilis. PMID- 6285489 TI - [Compressive pathology of ulnar nerve. Thirty seven cases reported (author's transl)]. AB - The authors have reviewed 37 operated cases of ulnar nerve compression, on the level of the elbow and the wrist, with a minimum follow-up of one year. Traumatic etiologies are more common on these two levels. The authors find 32% of pure entrapment syndromes at the elbow, whereas this phenomenon does not seem to exist at the Guyon's loge. Mac Gowan's classification was used to range the patients in three degrees, according to the importance of the palsy before operation. Surgical treatment was is all cases, at least decompression. At the elbow was added anterior transposition of ulnar nerve beneath epitrochleal muscles or beneath the skin. Considering the results, these lesions are all the more severe, as they are higher (elbow) and more advanced. PMID- 6285490 TI - [Radiological anomalies in the Noonan syndrome (author's transl)]. AB - In the Noonan syndrome various anomalies can be demonstrated radiologically. Of particular interest are cardiovascular lesions, such as atypical pulmonary stenosis, hypertrophic cardiomyopathy and the lymphangiectasias. Skeletal anomalies can be found, including retardation of bone maturation and sternal abnormalities. Other anomalies concern the urinary tract (pyeloureteral stenosis) and the digestive system. Radiological diagnostic procedures allow to disclose various elements of this polymalformative syndrome. They should be performed whenever suggestive clinical signs are present, e.g. hypertelorism, gonadal dysgenesis, down-slanting palpebral fissures and shield-like chest. PMID- 6285491 TI - [Spontaneous renal allograft rupture. Diagnosis and treatment. Report on five cases (author's transl)]. AB - The authors describe five cases of renal allograft rupture as a result of acute rejection. In one case of perineal hematoma, no operation was performed. The evolution of the hematoma led to a periureteral sclerosis, which was successfully operated. The authors advocate deferring the surgical management of allograft ruptures. Once the diagnosis has been established with the aid of physical examination, electrography and computerized scanography, the extra time can be devoted to following the evolution of the rejection. If the rejection is irreversible, a nephrectomy has to be performed, but if it is reversible, they favour conservative treatment followed by operative intervention around the tenth day. PMID- 6285492 TI - [Study of the spermatic bacterial flora in infertile males (author's transl)]. AB - Two groups of infertile males (65 and 132 patients) have been investigated in two different laboratories, with two different methods to obtain semen. The bacteriological results are quite similar in the two groups. The microorganisms which have been isolated are : beta-hemolytic Streptococcus, Proteus, Epidermidis staphylococcus, Micrococcus, Corynebacter, Viridans streptococcus, Klebsiella, Pseudomonas, Enterobacter, Bacillus, Neisseria, Escherichia coli, anaerobic Staphylococcus, anaerobic Streptococcus, anaerobic Corynebacter type IV. Fungus, Achromobacter, 20 p. cent of the patients are chronically infected without any clinical signs. This infection is probably of prostatic origin with an important number of bacteria in the semen (more than 3 000/ml). No relation has been shown between the bacteriological data and the physical and cytological characteristics of the sperm, except the pH : semens with a low pH are generally azoospermic and highly contaminated. PMID- 6285493 TI - [Bone and joint infection. Role of immune deficiences. Interest of serology and immunology in the diagnosis (author's transl)]. AB - Although bone and joint infections do not seem to be more frequent in patients with immune deficiences than in normal subjects, it seems paradoxically that an immune deficiency is relatively frequent during fully diagnosed bone and joint infections : the discrepancy between the two types of data is not easy to explain. Serology and immunology laboratories give little information in the etiological diagnosis of bone and joint infections, with the exception of perhaps gonococcal infections (search for anti-gonococcal antibodies by immunofluorescence). staphylococcal infections (pasteurella, yersinia, tularemia and brucella infections). In most cases, although the abnormalities observed are due to infection of the organism by the germ, they have nothing characteristic of the bone and joint localisation itself. PMID- 6285494 TI - [Aneurysmal cyst of the clavicle : an uncommon lesion and a difficult diagnosis (author's transl)]. AB - The authors describe a case of anevrysmal cyst of the clavicle seen in a five year-old girl. Both clinical and radiological diagnosis proved impossible. Complementary investigations did not settle the question. surgical excision was decided on. In spite of an unfavorable macroscopic aspect, histological examination determined the nature of the tumor. The authors emphasize the great value of resorting to all branches of medical investigation when dealing with bone tumors. Before affirming a good prognosis, or, on the opposite, suggesting extensive and mutilating surgery, the utmost caution is required. PMID- 6285496 TI - [Management of acute lymphoblastic leukemia in adults. Retrospective review of fifty case-reports (author's transl)]. AB - In adults, acute lymphoblastic leukemia (ALL) is less frequent and less clearly defined than in children. A retrospective study of fifty adult patients with ALL was carried out. Findings were similar to those reported in previous studies. Complete remission was achieved in 86% of patients. This rate is similar to that recorded in children with ALL. Median survival (twenty months) was shorter than in childhood because of frequent relapses. Various factors of prognostic value are discussed. Problems encountered in attempting immunological classification are studied. Prognosis seems to be related to the intensiveness of induction therapy and the choice of maintenance therapy regimen. PMID- 6285495 TI - [Inappropriate secretion of antidiuretic hormone (ISADH) after head injury (author's transl)]. AB - Two new observations of ISADH following head injury are described. A review of the medical literature is presented. Reports of ISADH after head injury are rare in comparison to the frequent occurrence of hydroelectrolytic disorders in the same situation. Attention is drawn to misleading clinical pictures, suggestive of neurosurgical conditions. Intracranial hematoma is frequently associated with ISADH and should be looked for in patients who fail to respond to therapy. PMID- 6285497 TI - [Immunofluorescence study of the distribution of ABH cell surface antigens in bladder tumours (author's transl)]. AB - ABH antigens are normally present at the surface of the healthy urothelium. In many cases of bladder tumours, the urothelium loses its antigens. While this loss would not seem to be associated with the grade or stage of the tumour, the persistence of the ABH antigens would seem to provide a more favourable prognosis. PMID- 6285498 TI - [Hemangiopericytoma. A review of eleven cases (author's transl)]. PMID- 6285499 TI - [Effects of local corticoids on the nerve fiber. Experimental study (author's transl)]. AB - The intra-operative establishment of some impairments of the median nerve, in patients who are operated for carpal tunnel syndrome and who previously received several local injections of corticoids, led the authors to try to see the respective responsibility of the compression and of the injection in the degenerative process. In 60- dog cubital nerves, it has been possible, to study different parameters by intra and perineural (recurrent or not injections: the puncture, the dilatation, the solvents, the benzylic alcool and two long action corticoids very often used (triamcinolone acetonide, cortivazol). In the first of our series, the injections were done around the nerve and inside the nerve under visual or blind control (as for an infiltration). The nerves are then cut off after 1, 2, or 3 months. In the second of our series, the injections were done in the same way, but recurrently on the first, second and third month, and the nerve was cut off within the fourth month. An electrophysiological study was performed before taking the samples; they were then prepared for reading on the optical and electron microscope. The results show lesions characterized by an epineural spumous sediment, a localized decrease in the large fibers under the perineurium in the first and second series. These impairments seem to be diminishing with time. However when the injections are done recurrently, the lesions are more important, showing a wallerian degeneration and regeneration. PMID- 6285500 TI - [Recurrent water intoxication during successive episodes of acute bronchitis: Schwartz-Bartter syndrome? (author's transl)]. AB - Mental confusion with hyponatremia and oliguria was recorded during each of three successive episodes of acute bronchitis. Antibiotics proved sufficient therapy and led to recovery. Thus, as in pneumonia, inappropriate secretion of antidiuretic hormone (ADH) may be seen in acute bronchitis. PMID- 6285501 TI - [Malignant systemic mastocytosis (author's transl)]. AB - The case of a 37 year old man with systemic malignant cutaneous mastocytosis is reported. The outcome was fatal after eight months. Death was due to a diffuse hemorrhagic syndrome which originated in an abnormal circulating heparinoid. Nosologically, this is a border-line case between mast cell leukemia and leukemic systemic mastocytosis. PMID- 6285503 TI - [Effects of gliclazide and glibenclamide on platelet function, fibrinolysis and metabolic control in diabetic patients with retinopathy (author's transl)]. AB - In order to compare the metabolic and hemobiological properties of two sulfonylureas, thirty-five non-insulin-dependent diabetics were randomly assigned to two groups. Each group was given either gliclazide (n = 20) or glibenclamide (n = 15) for six months. Metabolic control was improved in both groups, as evidenced by the decrease in HbA1 concentrations. A significant fall in ADP (1 and 4 microM)--induced platelet aggregation was recorded in the gliclazide group, while antithrombin III levels remained normal throughout the trial and plasminogen activator levels increased. These hemobiologic changes may be effective in preventing the vascular complications of diabetes. In contrast, glibenclamide did not alter platelet aggregation. Furthermore, a significant decrease in both antithrombin III levels and basal and venostasis-stimulated plasminogen activator levels were seen in glibenclamide patients. The beneficial changes in hemostasis seen in gliclazide patients probably result from a direct effect of the drug since hemobiological parameters and metabolic control parameters were not correlated. PMID- 6285504 TI - [Maternal glucose tolerance along pregnancy (author's transl)]. AB - The development of the foetal and placental unit induces large changes in maternal glucose tolerance along pregnancy. Oestrogen-induced hyperinsulinism is responsible for facilitated anabolism which take place during the first part of pregnancy. Accelerated catabolism occurring during the second part is due to the direct action of placental hormones, mainly of human placental lactogen. The latter is responsible for diminution of peripheral insulin activity. Hyperinsulinism, which is very important at this stage, facilitates an intense and rapid anabolism, mainly in the liver from where nutriments can be easily removed. Glucose and amino-acid uptake by placental and foetus are greatly increased by all these changes. PMID- 6285502 TI - [Cell mediated immunity in a mercury nephrotic syndrome (author's transl)]. AB - A case of extramembranous glomerulonephritis seen in a patient in contact with mercury diodide is reported. The responsibility of mercury in this occurrence of this nephropathy is confirmed by positive tests exploring cell mediated immunity. The other possible actions of mercury are described: modifications in renal tubular antigen, or in a tissue or serum protein, with antibody production and secondary apparition of immune complexes which deposit in the glomerules. PMID- 6285505 TI - [Thrombophlebitis of the superior sagittal sinus in patient with systemic lupus erythematosus (author's transl)]. AB - The case reported here is the first observation of thrombophlebitis of the superior sagittal and left transverse sinuses in a patient with systemic lupus erythematosus. Thrombophlebitis occurred thirty-six years after onset of SLE. The patient had neither nephrotic syndrome nor serum anti-coagulants. Various hypotheses concerning pathogenesis are suggested. The opportunity for heparin therapy is discussed. PMID- 6285506 TI - [Proliferative glomerulopathy in pneumococcal septicemia in a patient with lupus (author's transl)]. AB - Acute glomerulopathy due to pneumococcal infection occurred in a patient with chronic discoid lupus. The onset of renal involvement in this context suggested that cutaneous lupus may have developed into systemic lupus erythematosus. Renal biopsy established the correct diagnosis. The physiopathology of glomerular involvement in lupus and infections is discussed. Attention is drawn to the significance of complement activation, either by the classical or the alternative pathways according to the etiology. PMID- 6285507 TI - ["Deep scleroderma" or diffuse fibromatous scleroderma. A case-report of uncertain nosologic classification (author's transl)]. AB - An unusual form of scleroderma, seen in one patient, is described. Clinical and histological examinations demonstrated no dermal changes. Both the subcutaneous tissue and the fascias were involved. This case can be distinguished from Shulman fasciitis, on both histological and biological grounds, as well as from scleroderma, since Raynaud phenomena, sclerodactylia, and visceral involvement were lacking. PMID- 6285508 TI - [Value of superior vena cava angiography in Schwartz-Bartter syndrome (author's transl)]. AB - Schwartz-Bartter syndrome often results from lung cancer. With reference to two observations, attention is drawn to the value of opacifying the superior vena cava for demonstrating the tumor. This investigation also allows guided biopsy sampling as exemplified in the second observation. Vascular investigations seem all the more useful as small-cell carcinoma often develops outside the bronchi and are often inaccessible by endoscopy. PMID- 6285509 TI - [Xanthogranuloma of the lung: a tumor-like inflammatory lesion (author's transl)]. PMID- 6285510 TI - [Edematous forms of Horton arteritis. Two case-reports (author's transl)]. PMID- 6285511 TI - [Hypergammaglobulinemia and monoclonal gammopathy in Gougerot-Sjogren syndrome. Three case-reports. [author's transl)]. PMID- 6285512 TI - [Mediastinal involvement in primary amyloidosis with monoclonal gammopathy. Two case-reports (author's transl)]. PMID- 6285513 TI - [Serum immune complexes and cardiopulmonary bypass. A review of thirty-four cases (author's transl)]. AB - The immunologic status of patients undergoing cardiopulmonary bypass as investigated. Rheumatoid factor, cryoglobulinemia and serum immune complexes were looked for. Studies were performed before the operation and eight or fifteen days later. From the results, it is concluded that the immunologic changes that occur in the immediate postoperative period cannot be interpreted because of the profound modifications resulting from cardiopulmonary bypass. PMID- 6285515 TI - [Duodenal leiomyosarcoma with left hepatic metastasis treated by total exeresis (author's transl)]. AB - A case of duodenal leiomyosarcoma associated with a single liver metastasis successfully treated by gastro-duodenal resection and left lateral hepatic segmentectomy in one stage is reported. The result of hepatic resection for metastatic cancer depends on the location and the histological nature of the primary tumor, on the type of hepatic resection and on the number of metastatic deposits. The patient is alive 26 months postoperatively and there is no evidence of any recurrence. PMID- 6285514 TI - [Lathyrism and collagens I and III (author's transl)]. AB - The administration of lathyrogenic agents (BAPN) at weak and prolonged doses (9 weeks, 1 g/kg/day) induced in the rat simultaneous alterations of aortic and cutaneous connective tissues comparable to the modifications observed during ageing in man. The lathyrogens produced this effect of inhibiting lysine-oxydase, the enzyme responsible for cross-linking collagen and elastin which leads to an increase in extractable collagen in the tissues. But BAPN did not appear to affect synthesis of collagen either in amount or in types I and III collagen ratio. PMID- 6285516 TI - [An other case of Ogilvie's syndrome (author's transl)]. AB - The authors relate a case of acute functional obstruction, appearing after treatment of a cancer of rectum with extension to prevertebral plexus. The cause of the colic trouble appears to be neurological and due to a sympathetic and parasympathetic transmission block, while intrincic plexus was normal. Inefficacy of the contractions was established by radiology and endoscopy (electrical and pharmacodynamic stimulation and measure of pressure); it predominated in the left transverse colon. Caecal perforation has been avoided up till now after two years, without modification of clinical course, the patient evacuated his colon with the help of enemas. It seems probable that this case can be classified as Ogilvie's syndrome, the bibliography of which has been looked up and criticized. PMID- 6285517 TI - [The interest of CSF examinations in the etiological diagnosis of intracranial hypertension (author's transl)]. AB - The authors discuss the case of a 15-year-old girl suffering from intra-cranial hypertension. Neuroradiological examinations do not reveal any obstruction in the cerebral spinal fluid circulation. The CSF cylologic examination was the first to give etiological indications, showing the existence of malignant cells which the cerebral biopsy later enabled us to classify as glioblastoma. This case affords an opportunity to recall the three stages of the diagnosis: first, intra-cranial hypertension; secondly, chronic meningitis; thirdly, cytological difficulties in the examination of CSF in patients having undergone neuro-radiological examinations. PMID- 6285518 TI - Developmental aspects of alpha- and beta-adrenergic receptors. AB - Clearly a number of hormonal, pathologic, and maturation factors alter alpha- and beta-adrenergic receptors during development. Ontogenic changes in these receptors appear to be organ- and species-specific, occurring at distinct times during organ maturation. Developmental changes in adrenergic receptors are probably the result of a number of homologous and heterologous regulatory mechanisms. These are likely to include the effects of differentiation and aging themselves, and in fact some of the most dramatic changes in adrenergic receptor number and function occur during development. These changes are likely important to the adaptation of the developing infant and knowledge of their appearance, regulation, and function will be useful in predicting therapeutic or toxic effects of these treatments on the developing fetus and infant when exposed to adrenergic agonists or antagonists. PMID- 6285519 TI - Malignant fibrous histiocytoma of soft tissue with metaplastic bone and cartilage formation: a new radiologic sign. AB - The presence of bone and cartilage in some cases of malignant fibrous histiocytoma of the soft tissue as a microscopic finding has been reported previously but little note has been taken of the radiologic manifestations of these tumor elements. A series of five such cases with sufficient metaplastic osseous and cartilaginous elements to produce roentgenographic evidence of their presence is reported here. An additional two cases showed only histologic evidence of bone or cartilage formation. The reactive ossification tends to be peripheral in location, involving the pseudocapsule of the sarcoma or its fibrous septa. In three there was a zoning pattern with peripheral or polar orientation, strongly suggesting the diagnosis of myositis ossificans. The latter was the diagnosis considered radiologically in four of the five cases. Malignant fibrous histiocytoma with reactive bone and cartilage must be considered in the differential diagnosis of soft tissue masses with calcific densities, particularly when these occur in tumors of the extremities. PMID- 6285520 TI - Contamination of food plants and plant products with bacteria of public health significance. PMID- 6285521 TI - Dr. Moreno Canas: a symbolic bridge to the demedicalization of healing. AB - Costa Rica is a rapidly developing country with an expanding system of nationalized health care and a growing professional medical sector. This expansion affects the medicalization of health culture, including the symbolic and religious aspects of popular healing traditions. This study describes the myth and cult ot Dr Moreno Canas, a physician murdered in 1938, who is prayed to, or ritually invoked for cures and good fortune. The cult of Dr Morena Canas complements and modifies the healing role of the Costa Rican patron saint, the Virgen de Los Angeles, by idealizing modern science and technology. The cult of the doctor is a reaction of laypeople to the secularization of health problems, and to the medicalization of care. The myth and cult of Dr Morena Canas creates a popular image of secular medicine that laypeople sacrilize by treating it in a religious manner, and in doing so demedicalize control of their own care. PMID- 6285522 TI - [Functional tests in the diagnosis of thyroid cancer]. PMID- 6285523 TI - [Function of the pituitary-adrenal system in patients with brain injury subjected to conservative and surgical treatment]. PMID- 6285524 TI - [Role of the association of viruses and bacteria in the etiology of acute pneumonia]. PMID- 6285525 TI - [Clinico-morphological characteristics of primary liver tumors]. PMID- 6285527 TI - [Obstruction of the midportion of the respiratory tract in tracheal histiocytoma]. PMID- 6285526 TI - [Hepatitis A virus antibodies in children]. PMID- 6285528 TI - Defaecation frequencies in Black, Indian, Coloured and White populations - what do they signify? AB - Using questionnaires covering a 7-day period, frequencies of defaecation were determined in a total of 2937 Black, Indian. Coloured and White subjects aged from 10 years upwards. It transpired that the overall frequency in rural Blacks was roughly twice that of Whites. Among rural Blacks defaecation frequencies of 5 7 per week were recorded in 16%, and from 1-3 per day in 80%. The corresponding figures among urban Blacks were 25% and 71%, among Indians 58% and 34%, among Coloureds 46% and 39%, and among Whites 59% and 29%. The wide differential in frequencies between Blacks and Whites is consistent with the wide differential prevailing in proneness to noninfective bowel diseases. On the other hand, Indians and Coloureds had defaecation frequencies little different on average from those of Whites, yet the former two populations are far less prone to bowel diseases. Hence caution must be exercised in correlating defaecation frequency with proneness to bowel diseases. Since bowel behaviour includes numerous aspects, defaecation frequency should not be regarded in isolation. PMID- 6285529 TI - Kaposi's sarcoma in a renal allograft recipient with cytomegalovirus infection. A case report. AB - There are increasing reports of Kaposi's sarcoma arising in immunosuppressed patients, including renal allograft recipients. Furthermore, evidence is accumulating that cytomegalovirus infection may be an aetiological factor in Kaposi's sarcoma. We report an additional case in a renal allograft recipient treated with corticosteroids, azathioprine and niridazole, who also had active cytomegalovirus infection. PMID- 6285530 TI - Transfer of genes to Chinese hamster ovary cells by DNA-mediated transformation. AB - We have transferred DNa to Chinese hamster ovary (CHO) cells by DNA-mediated transformation. CHO tk- cells were transformed with the clones gene for herpes simplex virus thymidine kinase (HSV-tk) and were found to have a 50-fold lower frequency of transformation than mouse Ltk- cells at the same DNA dosage. By altering the amount of tk gene and carrier DNA present, frequencies of up to 5 x 10(-5) were obtained. CHO HSV-tk+ transformants were very stable, and in several clones the HSV-tk gene copies integrated in higher-molecular-weight DNA. These cells also exhibited cotransformation for unselected markers. CHO lines were also transformed at a frequency of 10(-4) with the bacterial gene Ecogpt in a SV40 pBR322 vector. CHO tk-cells could be transformed at a frequency of 10(-7) with cellular DNA isolated from CHO tk+ cells. CHO cells offer a well-defined genetic system within which to transfer either cloned or whole cellular DNAs. PMID- 6285531 TI - Mitochondrial DNA analysis of mouse-rat hybrid cells: effect of chloramphenicol selection on the relative amounts of parental mitochondrial DNAs. AB - Several mouse-rat somatic hybrid cell lines were isolated by fusing chloramphenicol-resistant (CAPr) and CAP-sensitive (CAPs) parent cells, and propagation of the parent mitochondrial DNA (mtDNA) species in the hybrid cells was studied. The restriction endonucleases EcoRI, HpaII, and HaeIII were used for identification of mtDNA species. Both mouse and rat mtDNAs were propagated in all the hybrid cells examined and maintained during long-term cultivation and repeated cell division. Moreover, in CAPr mouse-rat hybrid cells, selection and successive cultivation in the presence of CAP did not increase the relative amount of mtDNA species of CAPr parent cell origin, and when CAP was removed from the culture medium, mtDNA species of CAPr parent cell origin did not decrease appreciably. The amount of mouse mtDNAs was consistently 1-4 times that of rat mtDNAs inthe mouse-rat hybrid cells regardless of the species of parent cells from which the CAP resistance was derived. Thus mouse-rat hybrid cells have a stable mtDNA population in which the amount of mouse mtDNAs is larger than that of rat mtDNAs without any influence of CAP selection. PMID- 6285532 TI - Coronavirus 229E susceptibility in man-mouse hybrids is located on human chromosome 15. AB - Human coronavirus 229E, n enveloped, RNA-containing virus, causes respiratory illness in man and is serologically related to murine coronavirus JHM, which causes acute and chronic demyelination in rodents. 229E displays a species specific host range restriction whose genetic basis was studied in human-mouse hybrids. 229E replicated in human WI-38 cells but not in three mouse cell lines tested (RAG, LM/TK-, and A9). Human coronavirus sensitivity (HCVS) was expressed as a dominant phenotype in hybrids, indicating that mouse cells do not actively suppress 229E replication. HCVS segregated concordantly with the human chromosome 15 enzyme markers mannose phosphate isomerase (MPI) and the muscle form of pyruvate kinase (PKM2), and analysis of hybrids containing an X/15 translocation [t(X;15)(p11;q11)] localized HCVS to the q11 leads to qter region of chromosome 15. HCVS might code for a specific surface receptor, allowing 229E to be absorbed to and received within the host cell. PMID- 6285533 TI - Cyclosporin A in renal transplantation: a prospective randomized trial. AB - Cyclosporin A combined with prednisone was compared with standard immunosuppressive therapy (antilymphoblast globulin, prednisone, and azathioprine) in a prospective randomized trial of 100 mismatched, living related donor and cadaveric renal transplants. The results demonstrated cyclosporin A plus prednisone to be an effective immunosuppressive regimen for renal transplantation. The actuarial graft survival at 1 year was 93% for patients treated with cyclosporin A and 81% for patients treated with conventional immunosuppression. Patient survival was 98% for the cyclosporin A group and 100% for the conventional group. Cyclosporin A-treated patients had fewer rejection episodes and fewer infections complications including a marked decrease in the incidence of posttransplant cytomegalovirus infection. The side effects of cyclosporin A were mild, but nephrotoxicity caused by cyclosporin A was frequent and significant. Nephrotoxicity was reversible and managed by decreasing the daily cyclosporin A dose. It is concluded that the combination of cyclosporin A plus prednisone provides an excellent alternate immunosuppressive regimen for renal transplants as compared with conventional therapy. The consequences of long term administration of cyclosporin A are at present unknown. PMID- 6285534 TI - Aggressive multimodality therapy based on a multicompartmental model of glioblastoma. AB - Glioblastoma multiforme is composed of multiple cellular compartments with different morphologic, kinetic, metabolic, vascular, and genetic properties. Optimal therapy may consist of a variety of therapeutic strategies designed for individual compartments, administered in close temporal relation. These concepts may turn out to be valid for other solid tumors as well. Microwave-induced hyperthermia can be used to treat metabolically quiescent, relatively hypoxic, nondividing cells (Go) otherwise resistant to radiation and chemotherapy. Similarly, polychemotherapy can treat a broad spectrum of cell types if the blood brain barrier can be circumvented. Radical surgery, repetitively applied, can be safely used to "set up" experimental agents if the operation microscope and laser are employed. A consecutive series of 74 adult patients with malignant astrocytoma were treated with primary resection, radiation therapy, and 1,3, bis(2 chloroethyl) 1 nitrosourea chemotherapy. At recurrence, all patients were offered reoperation with the microscope and the laser prior to administration of phase-I agents--hyperthermia via an implantable miniature microwave antenna (6 cases); aziridinylbenzoquinone chemotherapy (13 cases); and blood-brain barrier reversal with dimethyl sulfoxide (DMSO) and polychemotherapy (9 cases). It was concluded that temperatures of 45 degrees C could be safely achieved and human tumors could not efficiently dissipate heat; that DMSO plus drug therapy could be tolerated but blood-brain barrier reversal demonstrated by us in animals could not be shown in humans; and that aggressive multimodality therapy and reoperation could produce a 40% 2-year survival rate for patients younger than 40 years. PMID- 6285535 TI - Long-term preservation of segmental pancreas autografts. AB - Clinical pancreas transplantation is logistically difficult because of uncertainty concerning how long pancreas grafts can be preserved. We studied the viability of canine segmental (tail) pancreatic autografts transplanted after 24 to 72 hours of hypothermic preservation in either modified Collins' solution or modified silica gel filtered dog plasma (SGF). All grafts stored for 24 to 48 hours functioned immediately (plasma glucose less than 140 mg/dl in recipients). Grafts that failed did so between 2 and 7 days, and five dogs died with functioning grafts. The long-term functional success rate was 80% for fresh transplants; 67% and 40% for grafts stored in Collins' solution for 24 and 48 hours, respectively; and 75%, 75%, and 30% for grafts stored in SGF for 24, 48, and 72 hours, respectively. If technical failures are excluded. 50% of grafts stored in Collins' solution and 100% of grafts stored in SGF for 48 hours functioned long term. K values for the intravenous glucose tolerance test at 2 weeks in dogs with functioning grafts ranged from -1.44% to -1.78% and were similar in all groups. In conclusion, pancreatic grafts can be preserved for 48 hours by simple cold storage, but SGF is more reliable than Collins' solution (P = 0.015). Four human pancreas grafts were stored in SGF for 7 to 22 hours, one with and three without prior warm ischemia, and all three of the latter functioned after transplantation. PMID- 6285536 TI - [Pharmacodynamics and therapeutic efficacy of cyto-poppy in patients with respiratory insufficiency]. PMID- 6285537 TI - [Sodium nitroprusside: mechanism of action and clinical application]. PMID- 6285538 TI - Infections in pregnancy: highlights from the collaborative perinatal project. AB - The longitudinal study of approximately 60,000 pregnant women was conducted at 14 major medical centers throughout the United States. Clinical data was obtained under prescribed protocols for each of the study women throughout their pregnancy, at delivery, and through the postpartum period. The children were followed with uniform examinations from birth through seven years of age. In addition blood samples were taken from the pregnant women at registration, every two months through pregnancy, at delivery, and six weeks postpartum. Cord blood samples were also taken from the children during the last half of the study. We have used this material from the Collaborative Perinatal Project for the investigation of the role of infections in the production of damage to the fetus and newborn. The frequency of clinically recognizing infection during pregnancy was determined and geographic variation was demonstrated. Serological tests were used to document certain diseases. The frequency of confirmed clinical cases per 10,000 were: rubella, 8; rubeola, 0.6; mumps, 10; varicella-zoster, 5. The epidemiology and clinical findings associated with infections were studied using serological methods. This has provided data on the frequencies of infections such as cytomegalovirus, herpes simplex, mumps, rubeola, respiratory syncytial virus, and others. The study of abnormal pregnancies and matched controls is in progress using serological techniques. A number of specific studies have been reported on infections including rubella, neonatal meningitis, cytomegalovirus, maternal urinary tract infections, and toxoplasmosis. Further testing is now in progress employing more sophisticated laboratory methods and more complete data analysis. PMID- 6285539 TI - Real-time scanning as an adjunct to conventional hepatic sonography. PMID- 6285540 TI - [Hormonal receptors of the prostate: the facts known today (author's transl)]. PMID- 6285541 TI - Effects of pulmonary function of whole lung irradiation for Wilm's tumour in children. AB - The effect of whole lung irradiation on lung function was investigated in 48 children treated for Wilm's tumour with pulmonary metastases. Lung function tests were performed before irradiation and were repeated annually for as long as possible, the length of follow-up varying from two to 17 years. A reduction in both lung volume and in dynamic compliance was clearly observed. In some patients these changes occurred in the early post-irradiation months, but in most the decrease observed progressed over longer periods of time. Static pressure volume curves, blood-gases, and carbon monoxide transfer were normal. These findings make it unlikely that post-irradiation pulmonary fibrosis was involved. Another explanation for the decreased lung volume and dynamic compliance might be failure of alveolar multiplication. Muscular injury is unlikely as the patients were able to produce normal transthoracic pressures. A failure of chest wall growth is also possible and would explain the progressive restrictive impairment but not the early lung function changes. It is suggested that the early effects detected in some patients were the result of lung injury and that later effects resulted from impaired chest wall growth. PMID- 6285542 TI - Lung angiotensin converting enzyme activity in rats with pulmonary hypertension. AB - We have studied serum and lung tissue angiotensin converting enzyme (ACE) activity in female Wistar rats with pulmonary hypertension induced by two different methods. Chronic pulmonary hypertension was produced in one group of 10 rats (CH) by confinement in a hypobaric chamber (380 mmHg) for three weeks, and in another group fo 10 rats (M) by a single subcutaneous injection of monocrotaline (60 mg/kg body weight). In these two groups of tests rats and in 20 untreated controls (C), we evaluated right ventricular mean systolic blood pressure (Prvs mmHg), right ventricular hypertrophy, and serum ACE (n mol/ml/min). In lung tissue homogenate, we measured the specific activity of ACE (n mol/mg protein/min), alkaline phosphatase (AP) (IU/mg protein) and lactic dehydrogenase (LDH) (IU/mg protein). The Prvs in groups, C, CH, and M was 25 +/- 7 SD, 41 +/- 7, and 51 +/- 5, respectively. The ratio of right ot left ventricular weight (RV/(LV + S)%) in groups, C, CH, and M was 29 +/- 4, 52 +/- 5, and 56 +/- 7, respectively. The lung tissue ACE in groups C, CH, and M was 85 +/- 11, 65 +/- 20, and 22 +/- 5, respectively. In groups CH, and M the Prvs and RV/(LV + S)% were significantly elevated above control values while lung ACE was significant decreased (p less than 0.05). There was a significant inverse relationship between lung ACE on one hand, and Prvs (r = - 0.73) and RV/(LV + S)% (r = - 0.71) on the other hand. Serum ACE and lung AP were unchanged. In group M there was a slight but significant reduction in lung LDH. Chronic pulmonary hypertension, irrespective of its method of production, is associated with decreased lung ACE. The reduction in lung ACE is inversely proportional to the severity of pulmonary hypertension and right ventricular hypertrophy. PMID- 6285543 TI - Effect of cAMP phosphodiesterase inhibitors on ADP-induced shape change, cAMP and nucleoside diphosphokinase activity of rabbit platelets. AB - The effects of cAMP phosphodiesterase inhibitors on ADP-induced shape change and cAMP concentrations have been studied. Caffeine (10 mM), theophylline (8 mM), dipyridamole (0.2 mM), or papaverine (0,05 mM) prevented the shape change of washed rabbit platelets induced by 0.4 microM ADP. At these concentrations, none of these cAMP phosphodiesterase inhibitors increased 14C-cAMP in platelets in which the cytoplasmic adenine nucleotides had been labelled with 14C-adenine. By a protein binding assay, only papaverine by itself increased platelet cAMP above its basal level. These results indicate that two pools of cAMP may exist in platelets. Both methods showed that stimulation of platelet adenylate cyclase with PGE1 (1 microM) resulted in an increase in platelet cAMP and all these cAMP phosphodiesterase inhibitors potentiated this increase caused by PGE1. By themselves, some of these compounds may act through mechanisms that do not involve platelet cAMP. The effects of these cAMP phosphodiesterase inhibitors on platelet nucleoside diphosphokinase (NDK) activity were also investigated. At concentrations that prevented ADP-induced shape change, papaverine and dipyridamole had no effect on the formation of 14C-ATP from 14C-ADP by washed rabbit platelets. The methylxanthines partially inhibited NDK activity of washed rabbit platelets and of isolated platelet membranes, probably due to the structural similarity between the adenine ring of ADP and these substances. However, adenine (8 mM) inhibited ADP-induced shape change and platelet NDK activity but was a less effective inhibitor of ADP-induced platelet aggregation. Thus it seems unlikely that interference with platelet NDK or the ADP receptor is the major mechanism by which the methylxanthines inhibit platelet functions. PMID- 6285544 TI - HLA antigens on glioma cells from short term cultures. AB - Biopsied tumor cells from astrocytoma-bearing patients were grown in primary culture for 3-5 days. Both low and high grade tumors were represented in the study. The cultured cells could be shown to express the HLA-A and -B antigens using a multispecific allo-antiserum and a rabbit anti-beta-2 microglobulin antibody. The tumor cells were negative for the HLA-DR determinants when tested with either a rabbit anti-Ia-like antisera or specific anti-HLA-DR allo-antisera. They also failed to stimulate allogeneic lymphocytes in primary mixed lymphocyte tumor cell cultures but moderately stimulated autologous lymphocytes from the tumor-bearing patient in most of the combinations tested. PMID- 6285545 TI - [Extramural care for the patient with cancer]. PMID- 6285546 TI - The follow-up study of swine and Hong Kong influenza virus infection among Japanese hogs. AB - Pigs in Miyagi Prefecture, Japan were examined for swine (Hssw1N1) and Hong Kong (H3N2) influenza virus infection by serological tests. The results obtained revealed that a swine influenza was prevalent with relatively high positive ratios throughout that time, and that the Hong Kong influenza virus closely related to a recent human epidemic strain, A/Yamanashi/2/77, also persisted, corresponding to a human endemic. These epidemiological findings strongly suggested the possibility of direct transmission of Hong Kong virus from humans to pigs and vice versa. PMID- 6285547 TI - Survival of and hepatoma development in patients with liver cirrhosis. AB - A follow-up study was made on 75 patients with liver cirrhosis. The patients were divided into 5 groups (discontinued alcoholic group, continued alcoholic group, hepatitis B group, male non-alcoholic non-hepatitis B group, and female non alcoholic non-hepatitis B group). Hepatoma developed in 50% of the hepatitis B group, but in 1-9% of the other 4 groups. Abstinence did not accelerate the development of hepatoma. As for survival after onset of liver cirrhosis, the 1-4 year survival of the female non-alcoholic non-hepatitis B group and the final (8 year) survival of the hepatitis B group appeared to be lower than that of the other groups. Though the survival of the discontinued alcoholic group was seemingly higher that that of the continued alcoholic group, the difference was small and not significant. PMID- 6285548 TI - Congenital secondary hypothyroidism with low serum GH and prolactin levels in a 27-day-old male infant. AB - A male infant with secondary hypothyroidism is described. Within the first month after birth, the patient manifested feeding difficulties, lethargy, persistent jaundice, umbilical hernia, and large anterior and open posterior fontanels. The roentgenogram of the knee joints at 27 days showed absence of the distal femoral epiphyses. His serum thyroid-stimulating hormone (TSH) level was low despite decreased levels of triiodothyronine (T3) and tetraiodothyronine (T4) in serum. Assessment of the hypothalamic-pituitary hormone (TRH) nor growth hormone (GH) responses to L-arginine and insulin, while responses of both luteinizing hormone (LH) and follicle-stimulating hormone (FSH) to luteinizing hormone-releasing hormone (LH-RH) and adrenocorticotropic hormone (ACTH) to insulin were within normal limits. The malady of the patient in this case was not detected by newborn screening for congenital hypothyroidism due to the fact that in the Aomori district of Japan thyroid screening involves only the measurement of TSH. Such measurement cannot detect cases of secondary or tertiary hypothyroidism such as our patient. Replacement therapy was initiated at 58 days and his physical and mental development has been regarded as normal since treatment. PMID- 6285549 TI - Plasma insulin response to various secretagogues in insulinoma. AB - Since the development of radioimmunoassay for insulin, the diagnosis of insulinoma has been made easily. However, it has been assumed that insulinoma is heterogenous in the histological structure as well as in clinical findings. Therefore, the present study was performed to investigate the insulin response to various stimuli and to evaluate the various insulin response tests in 19 patients with insulinoma. The fasting blood glucose was 19 to 90 mg/100 ml in insulinoma and 81 +/- 5 (mean +/- S.D.) mg/100 ml in normal controls. Plasma insulin (IRI) in insulinoma ranged from 10 to 255 microU/ml, while in the control it was 14 +/- 9 microU/ml. However, insulin/blood glucose ratio increased in insulinoma (0.2 11.2) compared with the normal control (0.18 +/- 0.11). In oral glucose tolerance tests, plasma IRI increased and reached peak levels of 48-244 microU/ml, remaining elevated in most cases. In the intravenous tolbutamide test, plasma IRI increased conspicuously to 82-1,330 microU/ml and hypoglycemic coma was provoked in 54%. Plasma IRI was elevated in the intravenous glucagon test and reached the peak levels of 85-400 microU/ml, which exceeded those of the control group. Plasma IRI increased to more than 100 microU/ml after arginine infusion and formed bizarre curves. There were no correlations between plasma IRI response to various stimuli and malignancy, type of B-granule or insulin content of insulinoma tumors. It is concluded that fasting plasma IRI, insulin/glucose ratio, tolbutamide test and glucagon test are highly valuable for the diagnosis of insulinoma. PMID- 6285550 TI - Acute effect of cigarette smoking on serum angiotensin-converting enzyme activity in normal man. AB - The acute effect of cigarette smoking on serum angiotensin-converting enzyme activity was evaluated in 6 healthy subjects consisting of 4 non-smokers and 2 habitual smokers. Cigarette smoking resulted in rapid increases in serum converting enzyme activity in 5 of 6 subjects within 5 min. and the converting enzyme activity remained above the control value at 30 min. The increase in the enzyme activity of non-smokers was higher than that of habitual smokers at any time when the enzyme activity was determined. It is therefore suggested that cigarette smoke (or smoking) can cause the secretion of angiotensin-converting enzyme from the pulmonary endothelial cell, in which the enzyme may be produced, to the systemic circulation. It is also speculated that the increase in the enzyme activity may contribute to the initiation of cardiovascular changes associated with cigarette smoking. PMID- 6285551 TI - The effect of diethyldithiocarbamate on brain uptake of cadmium. PMID- 6285552 TI - The effect of chelating agents on the excretion of endogenous metals. PMID- 6285553 TI - The cardiotoxicity of hydrogen cyanide as a component of polymer pyrolysis smokes. PMID- 6285554 TI - Isolation and identification of a collagenolytic enzyme from the venom of the western diamondback rattlesnake (Crotalus atrox). AB - A collagenolytic enzyme, with a molecular weight of 58,000 daltons and isoelectric point of 5.1, was isolated and purified from the venom of the rattlesnake Crotalus atrox by Sephadex G-100 gel filtration followed by chromatography on DEAE-Bio-gel A. The enzyme released alpha-chains from beta chains of the native collagen and cleaved in the helical region similar to other animal collagenases. The enzyme also hydrolyzed the PZ-peptide, however, it did not hydrolyze synthetic substrates for serine protease (such as TAME or ATEE). The enzyme had no hemorrhagic activity. Immunocross-reactivity suggested that only the venom from Crotalidae contain the enzyme. PMID- 6285555 TI - Dietary selenium, hepatic arylhydrocarbon hydroxylase and mutagenic activation of benzo(a)pyrene, 2-aminoanthracene and 2-aminofluorene. AB - The effect of dietary selenium (Se) on the hepatic aryl hydrocarbon hydroxylase (AHH) and mutagenic activation potential in young and adult rats was investigated. Animals were maintained on a low Se basal diet with or without 1 or 2 ppm Se supplementation (as sodium selenite) for 5 or 20 weeks. AHH inducibility was determined by an intraperitoneal injection of Aroclor 5 days prior to killing. A significant reduction in the AHH level was observed in low Se group of young untreated rats only. Dietary Se did not affect AHH inducibility by Aroclor treatment and the induced AHH levels were similar in both dietary groups. 5 weeks on low dietary Se did not alter the AHH levels in untreated or treated adult rats. The levels of Se-dependent glutathione peroxidase (GSH-P) were decreased more drastically in the young than adult rats by low dietary Se. Studies on the metabolic activation potential of liver enzymes from young rats on basal and Se supplemented diets showed that dietary Se did not alter the activation of benzo(a)pyrene (BP) to mutagens although differences in the activation of 2 aminofluorene (2AF) and 2-aminoanthracene (2AA) were observed. PMID- 6285556 TI - The epidemiology of Entamoeba histolytica in a rural and an urban area of Mexico. A pilot survey II. AB - Stocks of intestinal amoebae grown in monoxenic culture, were compared against each other and against those previously reported from Mexico City. These were isolated from subjects in San Cristobal de las Casas, Chiapas (rural area) and hospital patients in Merida, Yucatan (urban area). Electrophoretic patterns of the four enzymes: glucose phosphate isomerase (GPI), phosphoglucomutase (PGM), L malate: NADP+ oxido-reductase (oxalacetate-decarboxylating) (ME) and hexokinase (HK) demonstrated the presence of five groups (zymodemes) of Entamoeba histolytica already described from Mexico City, together with two new zymodemes, one of which gave a recognizable pathogenic pattern, whilst the other gave a contradictory pattern. Zymodemes of Entamoeba coli, Entamoeba hartmanni, Iodamoeba buetschlii and Dientamoeba fragilis, previously described were also isolated. One new zymodeme of D. fragilis was demonstrated. PMID- 6285557 TI - In vivo antileishmanial properties of pentavalent antimonial compounds. PMID- 6285558 TI - Fatal intravascular hemolysis associated with T-polyagglutination. PMID- 6285559 TI - Epidemiological studies on the animal reservoir of gambiense sleeping sickness. Part I. Review of literature and description of the study areas. AB - A review of literature of the previous and recent evidence of potential animal reservoir hosts of gambiense sleeping sickness is given. A highly endemic sleeping sickness area in the Ivory Coast is described together with an area of recent low endemicity in southwestern Upper Volta. These areas were surveyed as part of a study on the significance of any potential animal reservoir of the disease. The results are described in detail in subsequent papers. PMID- 6285560 TI - Epidemiological studies on the animal reservoir of gambiense sleeping sickness. Part II. Parasitological and immunodiagnostic examination of the human population. AB - The prevalence of the disease was determined in the population (n = 3402) of a highly endemic sleeping sickness area in the Ivory Coast and an area of recent low endemicity in southwestern Upper Volta by using parasitological techniques supplemented by the determination of trypanosome specific antibodies and serum macroglobulins (IgM). 62 cases of trypanosomiasis were diagnosed parasitologically in the sleeping sickness focus in the Ivory Coast. 30 Trypanozoon stocks were established for their behavioural and biochemical characterization. The percentage of specific antibody carriers. 26% and 10% obtained with the enzyme-linked immunosorbent assay (ELISA) in both areas, respectively, was compared to that obtained for raised IgM levels. False negative diagnosis resulted in 24% with the ELISA, 12% with the indirect immunofluorescent test (IFT) and 19% with the radial immunodiffusion (RID). Considering IFT and IgM levels together, the proportion of false negative results fell to 8%. Possible reasons for low sensitivities, especially of the ELISA are discussed. PMID- 6285561 TI - [The epidemiology of human trypanosomiasis due to Trypanosoma gambiense in a focus of the Ivory Coast. I. The distribution of the disease in the population (author's transl)]. AB - The authors have studied the distribution of trypanosomiasis in the population of the focus around Vavoua in the Ivory Coast. Because of numerous immigrants from Upper Volta who have settled in the area, the composition of the population differs from the general population of the Ivory Coast. The cases diagnosed from 1977 to 1979 were studied by village, ethnic group, sex and age. The study shows that the highest incidence of sleeping sickness occurs among men, in the age groups from 10 to 30 years and among immigrants from Upper Volta. PMID- 6285562 TI - Traps to control and estimate populations of Glossina species. AB - Biconical traps were used to capture continuously Glossina palpalis s.l. in Ivory Coast and G. morsitans centralis in Zambia for 19 and 20 days respectively. Both fly populations declined markedly during the continuous trapping period though populations subjected to intermittent trapping or where the traps were very widely spaced did not do so to the same extent. Fly density was estimated by applying the principles of removal trapping and were compared with other trapping studies. The results demonstrated that biconical traps removed 7% of the female component of the population each day and that this 7% removal value was itself a measure of absolute trapping efficiency; providing a novel means of population estimation. The potential of traps to control tsetse and thus break trypanosome transmission cycles or reduce 'challenge' on a self-help basis at the village level is discussed. PMID- 6285563 TI - [Immunodiagnostic findings in sera of patients with leishmaniasis, Chagas' disease, malaria and amebiasis in endemic regions of Venezuela (author's transl)]. AB - 66 serum samples from patients suffering from mucocutaneous leishmaniasis (20), Chagas' disease (12), malaria (16) or amebiasis (18) were collected and examined with five different protozoic antigens (L. donovani, T. cruzi, P. fieldi, P. falciparum, E. histolytica) by means of the indirect fluorescent antibody test, the complement fixation test, the indirect hemagglutination test and the latex agglutination. Cross-reactions were observed only between the sera from patients with leishmaniasis and Chagas' disease, both groups, however, showed stronger reactions with the homologous antigen. There were no cross-reactions among the other antigens and antibodies. The results obtained in this study together with facts already proved by literature show the usefulness of the employed antigens for epidemiological surveys. PMID- 6285564 TI - A nucleotidase from Leishmania tropica Promastigotes: partial purification and properties. AB - Nucleotidase activity which had been found in various subcellular fractions of Leishmania tropica has been partially purified from the supernatant of the Triton X-100 treated pellet of the 100 000 g centrifugation of the L. tropica homogenate by CM-cellulose column chromatography, Con A-Sepharose affinity chromatography, isoelectrofocusing and column chromatography on Sephacryl S-300. The enzyme showed the characteristics of a nucleotidase (EC 3.1.3.31) with a broad substrate specificity. It was inhibited by the products of the reaction and by alpha, beta methylene adenosine 5'diphosphate and adenosine 5'-9-thiomonophosphate. Nucleotidase activity of intact Leishmania cells and the inhibitory effect of Concanavalin A on the activity of the enzyme suggest also surface-associated properties of the enzyme. PMID- 6285565 TI - Further studies on the reinvasion of the onchocerciasis control programme by Simulium damnosum s.l.: the effects of an extension of control activities into southern Ivory Coast during 1979. AB - During 1979 the aerial larviciding operations of the Onchocerciasis Control Programme were extended southwestwards to cover those areas with Ivory Coast where the savanna cytospecies of the Simulium damnosum complex were known to breed. This resulted in a marked reduction of the reinvasion which threatened the Programme in Ivory Coast and Upper Volta in earlier years. Previous conclusions concerning the location of sources of invading populations and consequently the direction and distance of the movements of flies were substantiated. It was confirmed that S. damnosum s. str. and S. sirbanum were the main migrant species. In this area S. Squamosum was shown to be of much less importance as a migratory vector. PMID- 6285567 TI - Foot-and-mouth disease typing and serology in Zimbabwe. AB - A small foot-and-mouth disease diagnostic unit was established in Zimbabwe. Twenty-six out of 27 field specimens were successfully typed using the complement fixation technique. The antibody response in cattle to the SAT II vaccine was determined using a microtitre neutralisation method. Techniques are described which should be within the capacity of laboratories in developing countries. PMID- 6285566 TI - [Determination of the mutagenic potential of an environmental pollutant, mercuric chloride]. AB - The effect of mercury chloride on the culture of rat fibroblast cells was studied for the test of DNA-breaks formation, for the survival and mutability of pox virus vaccine and for the formation of dominant mutations in rats. The preparation had generally toxic, embryotoxic and mutagenic effects. To determine the genetic effects of environmental pollutants it was necessary to use some special methodological approaches. PMID- 6285568 TI - Goat pox in the Sudan. AB - Out outbreak of goat pox occurred among 60 adult and young goats in the Sudan. Among the findings of special interest was the extensive and wide distribution of pox lesions on the bodies of the animals. The severe involvement of the mucous membranes of the muzzle, eyes and nostrils gave rise to acute respiratory distress and systemic reaction. The suppression of secondary bacterial infection with local and systemic terramycin in addition to supportive therapy of affected animals alleviated the condition and none of the animals died. Typical pox lesions were produced experimentally on goats and sheep using infected material from goats. Human infections were not encountered. PMID- 6285569 TI - Congenital mesoblastic nephroma: comparative histological study of three cases at different ages. PMID- 6285570 TI - [Effect of vanadium compounds on the enzymic activity in sarcolemma of developing muscles]. AB - Sodium metavanadate (10(-4)-10(-6) M) stimulates the activity of adenylate cyclase and decreases the activity of Na+, K+-ATRase and 5'-nucleotidase in the sarcolemma fraction of chicken skeletal muscles at the embryonal and postembryonal developmental stages. Under conditions of a combined action of vanadate and guanylic nucleotides on the adenylate cyclase activity their effects are found to be potentiated. Epinephrine in vitro removes an inhibitory influence of vanadate on Na+, K+-ATPase from the third week of twe embryonal period. The restoring effect of epinephrine is blocked by propranol--a beta-adrenoblocker. PMID- 6285571 TI - [Triiodothyronine binding and norepinephrine uptake in the heart and liver]. AB - Administration of norepinephrine to thyroidectomized rats activates sharply the [125I]triiodothyronine binding by heart mitochondria and liver nuclei. Epinephrine stimulates the binding by the heart mitochondria and decreases the intensity of this process in the liver and heart nuclei and liver mitochondria As compared to norepinephrine, adrenoxyl is weaker in activation of [125I] triiodothyronine binding by the heart mitochondria and stronger in intensification of binding by the liver nuclei. Physiological concentrations of thyroxine like adrenoxyl administered to intact animals 2h before investigations intensify the uptake of [3H] norepinephrine by sections of the auricles, myocardium and liver. Hyperthyroidization induces contrary changes inthe uptake of [3H] norepinephrine. Norepinephrine administration decreases sharply the uptake of [3H] norepinephrine by sections of the auricles and myocardium. The blocking of beta-adrenoreceptors weakens the uptake. PMID- 6285572 TI - Unusual intranuclear inclusions in malignant fibrous histiocytoma: presence in primary tumor, metastases, and xenografts. AB - We describe previously unreported intranuclear inclusions in 2 cases of malignant fibrous histiocytoma. The inclusions were found in 2-10% of the tumor cells removed from the patients and in 2-10% of the cells examined in tumor tissue xenotransplanted in nude mice. By stereo electron microscopy the inclusions are closely packed undulating fibrils 18-23 nm in diameter. They are sometimes associated with fibrillary bodies. They closely resemble the inclusions reported in some animals inoculated with serum from patients with non-A non-B hepatitis; however, their nature at present is unknown. PMID- 6285573 TI - Intracellular structures in an astrocytoma. PMID- 6285574 TI - [Endorphins and opiated receptors]. PMID- 6285575 TI - Acute adrenal insufficiency: recognition, management, and prevention. AB - Acute adrenal insufficiency may present only with nonspecific symptoms and signs. Hyperpigmentation is not a feature of secondary adrenal insufficiency and is absent in patients with primary adrenal failure of recent or acute onset. Similarly, characteristic electrolyte disturbances may be obscured by concomitant vomiting and diarrhea as well as by parenteral electrolyte replacement. A high index of suspicion must therefore be maintained to make the diagnosis of acute adrenal insufficiency in patients without a recognized history of autoimmune adrenal insufficiency or of other diseases or therapeutic regimens known to result in pituitary-adrenal failure. Timely intervention with volume and glucocorticoid replacement rapidly reverses all symptoms and signs of adrenal insufficiency. Guidelines are presented for glucocorticoid replacement in the treatment of adrenal crisis as well as for the prevention of acute adrenal insufficiency in patients with known or suspected pituitary-adrenal disease. In addition, recommendations are given for the simultaneous diagnosis and treatment of adrenal insufficiency in patients without previously established disease. PMID- 6285576 TI - Complications of uremia. PMID- 6285577 TI - [Hormone content of the blood in Meniere's disease and cochleovestibulopathy]. PMID- 6285578 TI - [Patterns in the development of cancer of the gastric stump]. AB - The article analyzes results of the examination and surgical treatment of 24 patients with carcinoma of the gastric stump developing at different terms after resection of the stomach for gastroduodenal ulcers. Questions of pathogenesis, clinical picture and diagnosis of the disease are discussed. Special attention is called to low percentage of radical operations for gastric stump carcinoma. An important role of dispensary observation with the use of gastroscopy for patients subjected to resection of the stomach for peptic ulcers is emphasized. PMID- 6285579 TI - [Neurogenic stomach tumors]. AB - Preoperative diagnosis of neurogenic tumors of the stomach is known to be difficult and should be performed in complex with clinico-roentgenological and endoscopic methods. Among the most dangerous complications of neurogenic tumors of the stomach are hemorrhages, malignization, stenosis and abscesses. The neurogenic tumors of the stomach must be treated only surgically. The operation of choice is resection of the stomach. In cases with small benign tumors a resection of the tumor and part of the gastric wall is possible. PMID- 6285580 TI - [Radioisotope diagnosis of the hepatic changes in biliary tract diseases]. AB - The authors analyze the application of the radioisotopic method for examining the liver in 413 patients; with acute cholecystitis (97), chronic cholecystitis (195), chronic pancreatitis (12) and cirrhosis of the liver (109). The scanning and functional investigations of the liver allowed detection of substantial disorders in the functional state of the liver in patients with acute cholecystitis and obturation jaundice which suggests a necessary correction of the liver function in the pre- and postoperative periods. PMID- 6285581 TI - The use of corticosteriods for treatment of neurologic disease. PMID- 6285582 TI - [Analysis of the susceptibility of the Minor inbred chicken strain to Rous sarcoma virus infection]. AB - The sensitivity and resistance of the Minor hen inbred line to two antigenic subgroups of the Rous sarcoma virus were determined by the method of the infection of chorioallantoic membranes. The Minor line is sensitive to infection with the virus of the antigenic B subgroup and, as demonstrated by the testing of the interline hybrids of F1 and B1 generations, the birds of the Minor line are dominant homozygotes of genotype bsbs. The testing with the virus of antigenic A subgroup revealed a heterogeneity of the inbred population as to sensitivity and resistance. A formula was derived for the determination of the frequency of the dominant allele as determining sensitivity. PMID- 6285583 TI - [The effect of sodium nitrite on rats infected with avian sarcoma virus]. AB - A potential effect of sodium nitrite was studied as exerted on carcinogenesis elicited by the avian sarcoma virus in the body of rats. In the case of the infection of the rats with indefective avian sarcoma virus B77 (Bratislava strain; ID50 10-25/0.1 ml) and i. p. inoculation of sodium nitrite, great mortality occurred already on the 20th day p. i. On the 125th day p. i. the cumulative mortality in this group was 76.92%; on the other hand, in the control groups (rats inoculated with sodium nitrite or avian sarcoma virus) no deaths occurred before this period. The rats infected with a double dose of the virus were not observed to show pronounced changes in mortality. It was only on the 125th day that cumulative mortality in the group of rats infected with the avian sarcoma virus and rats inoculated s. c. with sodium nitrite was 87.5% and in the group of animals which had only been infected with the virus the mortality was 64.26%. The patho-morphological changes were in keeping with the hitherto described changes in rats infected with the avian sarcoma virus. In the trial no effect of sodium nitrite on the patho-morphological changes induced by the avian sarcoma virus was observed. PMID- 6285584 TI - [The effect of feeding clinoptilolite on the health status, blood picture and weight gain in pigs]. AB - Under experimental conditions, the effect of the administration of 5% the Nizny Hrabovec locality was studied as exerted on the health condition, blood picture, weight gains, faeces production and odour in pigs and on the course of gastroenterites of alimentary origin affecting these animals. The administration of mixture A1 with a 5% zeolite supplement did not exert any undesired influence on the white or red blood picture and on the general condition of health; on the contrary, weight gains increased by 23% as compared with the controls. The faeces of the treated animals were normally shaped and the odour was just slight. The administration of zeolite to the animals with gastroenteritis had a favourable influence on the health condition and weight gains. Clinoptilolite as a non traditional additive is a suitable means of improving nutrient conversion, of increasing the weight gains, of controlling diarrhoeas of alimentary origin and eliminating odour in large pig houses. PMID- 6285585 TI - Suspected ammonium nitrate fertiliser poisoning in cattle. AB - The occurrence of a suspected mixed ammonia and nitrate fertiliser poisoning that led to acute illness and fatalities in cattle is described. The circumstances leading to the incident included the method of application of the fertiliser, low rainfall and the absence of subsequent irrigation of the fertilised pastures. The clinical and gross post mortem findings, principally dehydration and fluid distension of the rumen, were not pathognomonic. The complex of nitrate, nitrite and ammonia toxicity is discussed. PMID- 6285586 TI - Interaction of rotavirus and enterotoxigenic Escherichia coli in conventionally reared dairy calves. AB - A study was made of the effects of rotavirus and/or enterotoxigenic Escherichia coli (ETEC) on dairy calves born and suckled on the farm and subsequently reared in isolation. Calves were orally inoculated at 6 days old with either rotavirus (5), ETEC (7), rotavirus and ETEC (5) or remained uninoculated controls (4), and their reactions were recorded by clinical, microbiological, and pathological observations. Rotavirus infection consistently produced diarrhoea, while ETEC inoculated alone did not colonise the intestine. In dual infections, both rotavirus and ETEC multiplied, although the severity of diarrhoea was not greater than that caused by rotavirus alone. Some ETEC-inoculated calves developed subsequent naturally-acquired rotavirus infections, but in these no ETEC multiplication occurred. The results suggest that prior or simultaneous rotavirus infection is necessary to enable ETEC colonisation of the intestine in conventional calves of this age. PMID- 6285588 TI - Rotavirus infections associated with diarrhoea in calves in Egypt. AB - The successful isolation and identification of rotavirus from newborn calves with diarrhoea is reported for the first time in Egypt. From 25 faecal samples taken from diarrhoeic calves, ten virus isolates were found to give cytopathogenic effects on bovine embryonic kidney cells. Three of the isolates were identified as rotavirus using fluorescent antibody staining, serum-neutralization, revealed the presence of rotavirus antibodies in 18 of 105 serum samples obtained from other calves slaughtered at Cairo abattoir. PMID- 6285587 TI - The zoonotic potential of bovine leukemia virus. AB - Many workers have investigated the possibility that bovine leukemia virus (BLV) might be transmissible to man. The epidemiological studies were designed to examine for associations between human leukemia and a rural environment, cattle farming, veterinary activities, or bovine leukosis. The serological studies were used to test serum samples from human cancer patients and from persons with potential occupational exposure to BLV, among others, for evidence of BLV antibodies. All these studies are critically reviewed. It is concluded that there is no epidemiological or serological evidence from human studies to indicate that BLV can infect man. PMID- 6285590 TI - Cytoplasmic inclusions in lymphocytes of chronic lymphocytic leukaemia. A report of 10 cases. AB - Peripheral blood from 90 CLL patients was examined by light-and electron microscopy for the occurrence of crystalline inclusions in lymphocytes. Inclusions were demonstrated in 10 patients (11%). In these patients the inclusions were present in 5-45% of peripheral blood lymphocytes. In the light microscope the inclusions appeared as rectangular, unstained structures in May Grunewald Giemsa and PAS stains. In the electron microscope the inclusions appeared as intracytoplasmic, completely partially membrane-bound bodies, which were often associated with dilated profiles of rough endoplasmic reticulum. The ultrastructure of the inclusions was granular. In immunofluorescence staining the inclusions were found to contain immunoglobulin of the same type and class as the surface membrane-bound immunoglobulin of the neoplastic lymphocytes, most frequently IgM-lambda. The lymphocytes of one case with kappa light chains at the cell surface membrane contained inclusions of the same ultrastructural morphology as those of the other cases with lambda light chains. The presence of inclusions was not associated with any specific clinical or prognostic features. the inclusions persisted during antileukaemic therapy. Their formation may be related to a dysfunction in the synthesis of surface membrane-bound immunoglobulins. PMID- 6285589 TI - [Comparative studies of plaque formation by strains of the mucosal disease-viral diarrhea virus (MD-VD)]. AB - Studied was the phenomenon of plaque forming with 2 local and 1 reference cytopathogenic strain of the MD-VD virus in cell cultures of calf kidney and testis. Studied was also the effect of trypsin in the preliminary treatment of the cell culture as well as the varying composition of the cover medium. The strains were found to form plaques that were varying in type and size. The ratio between the large and small plaques as well as the percent of plaques having unclear edges could serve as tentative strain features. It was established that at such experimental procedure the preliminary trypsin treatment of the cultures increased but slightly the plaque-forming capacity of the strains. The results obtained give grounds to the necessity of characterizing the vaccinal strains used through the type and ratio of the plaques formed. PMID- 6285592 TI - Cloning of uvsW and uvsY genes of bacteriophage T4. PMID- 6285591 TI - Association of SV40 large tumor antigen and cellular proteins on the surface of SV40-transformed mouse cells. PMID- 6285593 TI - Characterization of P1argF derivatives from Escherichia coli K12 transduction. II. Role of P1 in specialized transduction of argF. PMID- 6285594 TI - An examination of adenovirus type 5 mutants for their ability to induce group C adenovirus tumor-specific transplantation antigenicity in rats. PMID- 6285595 TI - Productive infection of embryonal carcinoma cells with ecotropic mouse type C viruses and subsequent arrest of differentiation. PMID- 6285596 TI - Infection of the human T-cell-derived leukemia line Molt-4 by Epstein-Barr virus (EBV): induction of EBV-determined antigens and virus reproduction. PMID- 6285597 TI - Properties of the genome of equine herpesvirus type 3. PMID- 6285598 TI - Characterization of monospecific antisera against all five vesicular stomatitis virus-specific proteins: anti-L and anti-NS inhibit transcription in vitro. PMID- 6285599 TI - Viral interference of HSV-1: properties of the intracellular viral progeny DNA. PMID- 6285600 TI - Topography of phosphate residues in Sendai virus proteins. PMID- 6285601 TI - Increase in the saturation of C18 fatty acids induced by coxsackie B6 virus in Vero cells. PMID- 6285602 TI - Interference grouping of murine leukemia viruses: a distinct receptor for the MCF recombinant viruses in mouse cells. PMID- 6285603 TI - Analysis of Yaba tumor poxvirus-induced proteins in infected cells by two dimensional gel electrophoresis. PMID- 6285604 TI - A novel cell transformation with DNA-containing cytoplasmic Yaba tumor poxvirus. PMID- 6285605 TI - Isolation and genetic characterization of temperature-sensitive mutants of simian rotavirus SA11. PMID- 6285606 TI - Persistent infection by a temperature-sensitive mutant isolated from a Sendai virus (HVJ) carrier culture: its initiation and maintenance without aid of defective interfering particles. PMID- 6285607 TI - Formation of a nucleosome-free region in SV40 minichromosomes is dependent upon a restricted segment of DNA. PMID- 6285608 TI - Microscopy of internal structures of Sendai virus associated with the cytoplasmic surface of host membranes. PMID- 6285609 TI - Superimmunity: characterization of a new gene in the immunity region of P1. PMID- 6285610 TI - Changes in the expression of Fc receptor produced by induction of Epstein-Barr virus in lymphoma cell lines. PMID- 6285611 TI - Class II defective avian sarcoma viruses: comparative analysis of genome structure. PMID- 6285612 TI - Tandem integration of complete viral genomes can occur in nonpermissive hamster cells transformed by linear BKV DNA with cohesive ends. PMID- 6285613 TI - Conserved polyadenylation signals in two negative-strand RNA virus families. PMID- 6285614 TI - [Effect of prostaglandin E2 on energy metabolism in isolated rat liver mitochondria]. AB - Prostaglandin E2 at concentrations of 5.6 mM and 56 mM, in presence of EDTA 2.2 mM, did not affect the patterns of respiration and coupling, registered by means of polarographic procedure, in isolated rat liver mitochondria. Activation of phosphorylating and uncoupled respiration of mitochondria was noted in incubation mixtures containing EDTA 2.2 microM, Ca2+ 0.2 microM, prostaglandin E2 5.6 microM. In absence of EDTA prostaglandin E2, at concentrations of 5.6 microM and 56 microM, caused uncoupling of the oxidative phosphorylation and in presence of Ca2+--it caused uncoupling and inhibition of mitochondrial respiration, decrease in the rate of phosphorylation and in the ATPase reaction was well as in the efficiency of mitochondrial proton pump, connected with Ca2+ transport. At concentrations 2.8 microM, 5.6 microM, 14 microM protaglandin E2 increased the proton conductivity of the mitochondrial membrane in presence of K+ and valinomycine. The data obtained suggest that the effect of prostaglandin E2 on the energy functions of mitochondria depends on presence of Ca2+ in the incubation mixture. PMID- 6285615 TI - [Regulatory enzymopathies]. AB - Hereditary regulatory enzymopathies are considered, molecular-genetic mechanism of which involved a mutation of a structural gene in the locus, coding the much less than regular much greater than amino acid sequence in the allosteric site of regulatory enzyme or the site of the enzyme polypeptide chain, which is responsible for allosteric conformational transition. Dissimilar manifestations of regulatory and classical enzymopathies are discussed. Estimation of kinetic patterns might be very useful in diagnosis of regulatory enzymopathies as shown during study of some molecular pathologies, which occurred, for example, due to molecular pathology of phosphofructokinase and phosphoribosyl pyrophosphate synthetase. Impairment of allosteric regulation of L-threonine-L-serine dehydratase is considered as a model of regulatory enzymopathy, found in spontaneous hepatomas of highly carcinogenous CBA mice strain. Treatment of regulatory enzymopathies is considered depending on the effect of well known chemotherapeutic drugs (mainly, acetyl salicylic acid) on allosteric functions of regulatory enzymes acetyl-CoA-carboxylase and fructose-1,6-bisphosphatase from animal liver tissue. PMID- 6285616 TI - [Activities of succinate dehydrogenase and ATPase in rat kidneys during transport of organic substances]. PMID- 6285617 TI - [Characteristics of 35S-lipoic acid absorption by the blood cells in breast cancer]. AB - The peculiar characteristics of 35S lipoic acid uptake by blood red cells in breast cancer patients were studied. The rate of 35S lipoic acid uptake by red cells in cancer patients was much higher than that in healthy female donors. The difference was less significant in cases of acute mastitis or benign tumors. The increased uptake of 35S lipoic acid by red cells may prove instrumental in the diagnosis and, probably, therapy of breast cancer. PMID- 6285618 TI - [Clinico-biochemical characteristics of chronic ischemic heart disease]. PMID- 6285619 TI - [Effect of preparations increasing the antiblastic action of antitumor substances on the results of postoperative chemotherapy in lung cancer]. PMID- 6285620 TI - [Prolactin secretion in diseases of the thyroid]. PMID- 6285621 TI - Mullerian adenosarcoma of the uterus. PMID- 6285623 TI - [Small cell carcinoma of the lung (author's transl)]. AB - Of the various types of bronchogenic carcinoma small cell carcinoma has the poorest prognosis. It is the most malignant of the histological subtypes of lung cancer owing to its rapid growth and early, widespread metastases. Small cell carcinomas are radiosensitive and respond to cytotoxic therapy. In the Second Surgical Department of the Vienna University Hospital, 70 "early-stage" small cell carcinomas were resected between 1955 and 1974. 27.5% of the patients with stage I and II carcinomas survived for more than 5 years without symptoms. "Curative" surgery is selected cases leads to prolonged survival in patients with localized lesions. PMID- 6285622 TI - [Prevention of recurrence of small-cell lung cancer by adjuvant polychemotherapy (author's transl)]. AB - No significant further improvement can be awaited in the results of purely surgical treatment of bronchial carcinoma. As from 1969--after preliminary studies--patients with bronchial carcinoma were treated by radical surgery and subsequently assigned within the framework of a randomized cooperative study, to a three-year course of intermittent high-dose polychemotherapy (cyclophosphamide, 5-FU, MTX and Velbe). A comparison according to the life-table method of the death rate of patients with small-cell bronchial carcinoma treated by combined polychemotherapy after surgery vis-a-vis those who received no polychemotherapy shows a rise of about 15% in the 4-year survival rate. This clear-cut improvement in survival rate emphasizes the fact that the radical operation is also indicated in these tumour forms and that the results of high-dose polychemotherapy are improved with this particular combination. With a view to confirmation and further improvement, a large international working group was established and has already submitted initial positive results concerning the applicability of a new alternating chemotherapeutic schedule. It is pointed out that the initial impetus towards undertaking such studies was given by Prof. Dr. Dr. h.c. Wolfgang Denk already in 1954 and that the consistent continuation of this combined inter disciplinary approach to the therapy of bronchial carcinoma is founded on his scientific foresight and organisational achievements. PMID- 6285624 TI - [Importance of steroid receptors for hormone therapy in advanced breast cancer]. PMID- 6285625 TI - [New findings on the metabolism and importance of the D vitamins, with special reference to the use of vitamin D]. AB - Animal-experimental examinations show that the peroral or intramuscular application of a high dose of vitamin D2 or of D3 leads to a toxic effect of these compounds on the osteocytes and that the hypercalcaemia evoked by this is mainly to be traced back to an increased deliberation of calcium from the bones. After application of a larger dose of vitamin D the activation mechanism in the liver and in the kidneys is much inhibited for several weeks so that no formation of 1,25-hydroxy-vitamin-D takes place; consequently, no furthering effect on the mineralisation of the bones is performed. Therefore, it is recommended to use physiological doses in the prevention of rachitis (500-1,000 IU a day). During the pregnancy the activity of the enzymes which participate in the activation of the D-vitamins increases in the liver and the kidneys. The kidneys of the fetuses are able to form 1,25-hydroxy-vitamin-D. Vitamin D and 25-hydroxy-vitamin-D transgress through the placenta into the fetuses. Due to the adaptation mentioned and the increased formation of 1,25-hydroxy-vitamin-D the absorption of calcium and phosphate increases during pregnancy. Recent pathobiochemical knowledge concerning the metabolism of the D-vitamins in several diseases are described. PMID- 6285626 TI - [Clinico-parasitologic basic diagnosis of extraintestinal amebiasis and evaluative comparison of different methods in the diagnosis and course of amebic liver abscess]. PMID- 6285627 TI - [Constipation in the aged--diagnosis and therapeutic aspects]. AB - The normal stool weight is 121 +/- 25 g per day with a stool frequency of 1.1 +/- 0.9 defecation per day. A normal transit time in man of 66 +/- 23 hours has been found. These data are necessary for the diagnosis of constipation. A recommendation of a fibre-rich diet or a purified plant fibre for therapy has been made. The dosage of bassorin or bran is 0.5 g per kg/day. In older people a drug therapy can often be necessary. PMID- 6285628 TI - [Diverticulosis-diverticulitis]. AB - Diverticulosis of the colon and its clinical sequelae--diverticulitis, peridiverticulitis, and pericolitis--are typical diseases of elderly people. The main causes are weakness of the connective tissue in the colonic wall increasing with age and pathologic motility pattern of the colon due to low dietary fibre consumption. In about 20% of all people with diverticula, acute or chronic recurrent diverticulitis develops, often with serious complications as perforation, abscess or fistula formation, obstruction, inflammatory pseudotumor and intestinal bleeding. Diagnosis is mainly based on clinical examination and barium enema (double contrast, maximal spasmolysis). Colonoscopy may be helpful in excluding carcinoma of the large bowel. Patients with diverticulosis and uncomplicated diverticulitis should be managed conservatively by medical treatment. The following measures proved to be successful: high-fibre diet, unprocessed wheat bran, and hydrophilic plant colloids to regulate the bowel movements, systemic or local antibiotics if signs of inflammation are present, and antispasmodics or analgesics against abdominal pain. Prognosis depends mainly on the duration of the disease, sufficient dietary-fibre intake, and elective or semi-elective surgical intervention before the development of life-threatening complications. The question as to whether diverticula or relapsing attacks of diverticulitis can be prevented with added dietary-fibre remains open for the time being. PMID- 6285629 TI - [Colonic polyps]. PMID- 6285630 TI - [Biochemical pharmacology of cardiac glycosides]. PMID- 6285631 TI - [Trends in experimental pharmacology]. PMID- 6285632 TI - [Pharmacology of purinergic systems]. PMID- 6285633 TI - [Current problems in pharmacology of the kidneys]. PMID- 6285635 TI - [Analysis of the inorganic part of tooth enamel using the electron paramagnetic resonance method]. PMID- 6285634 TI - [Electron paramagnetic resonance and its use in dental research]. PMID- 6285636 TI - [Surgical treatment of syndactylia in recessive dystrophic epidermolysis bullosa (author's transl)]. AB - The authors report on their experience with the surgical treatment of syndactylia and flexor contracture in recessive dystrophic epidermolysis bullosa in a 7 1/2 year-old boy. After separation of the fused fingers by excision of the epidermal sac, all sclerotic tissue and affected skin, the defects are covered by split autograft skin taken from clinically non-affected areas. Morphologic and functional results are very satisfactory. PMID- 6285637 TI - Demonstration of C1q - binding to collagen - anticollagen immune complexes in synovial fluids of RA-patients. AB - Collagen-anticollagen immune complexes were demonstrated indirectly in RA synovial fluids by digestion with collagenase and subsequent antibody determination. Elimination of C1q was performed by affinity chromatography using an anti-C1q column. Binding of C1q to collagen-anticollagen complexes was proven by demonstrating that collagenase-caused liberation of collagen antibodies was abolished or distinctly reduced after passage through the anti-C1q column. Binding of C1q to collagen-anticollagen immune complexes was proven via the fixation of these aggregates onto the anti-C1q column. This was achieved in those 7 RA synovial fluids which contained distinctly demonstrable collagen anticollagen complexes. PMID- 6285638 TI - Influence of a glycosaminoglycan polysulfate (Arteparon) on lysosomal enzyme release from human polymorphonuclear leukocytes. AB - In order to study the mechanism of action of a sulfated glycosaminoglycan (Arteparon) its influence upon the release of lysosomal enzymes from human polymorphonuclear leukocytes (PMNL) was investigated. Several fractions of tested glycosaminoglycan were used, differing in molecular weight (from 3,000 to 17,000). Human PMNL were incubated with zymosan particles opsonized with autologous serum. Preincubation with separate glycosaminoglycan fractions in concentrations 10(-3) M and 10(-4) M for 1 hr resulted in a significant activity decrease of released beta glucuronidase, neutral proteinase and myeloperoxidase. The concentration 10(-5) M was less active. Determination of total enzyme activities were made after cells (either incubated with tested glycosaminoglycan fractions for 1 hr or without incubation) were lysed by six freeze-thaw cycles. Sulfated glycosaminoglycan fractions inhibited all total enzyme activities. These results suggest direct inhibitory effect of tested drugs on lysosomal enzymes based on an interaction between inhibitor and enzyme. PMID- 6285639 TI - The effect of dietary composition on transit time in rats. PMID- 6285641 TI - Activity of enzymes in organic carbon-amended soils treated with varying levels of salts. AB - The effect of varying concentrations of either NaCl or Na2CO3 (0, 10, 20, and 25 meq/100 g soil) and organic carbon (0 and 2% starch) on the activity of dehydrogenase, urease, and phosphatase (nuclease) was studied in incubated samples of alluvial clay and calcareous sandy loam soils. Moisture content was kept at 60% W.H.C. The level of 10 meq/100 g soil of either sodium chloride or sodium carbonate was stimulatory for the activity of the three enzymes studied in both soils tested. The increasing concentrations of Na2CO3 showed greater changes in the enzymatic activity than the corresponding concentrations of NaCl in both soils. Application of starch reduced the inhibitory effect of the high levels of such salts on the enzymatic activities in both soils, except for phosphatase which was depressed by Na2SO3 in starch-amended soil samples. The calcareous soil responded to the starch addition less than the alluvial soil. PMID- 6285640 TI - [Effect of various rations on trace element metabolism in young fattening bulls]. PMID- 6285642 TI - Enzymatic activities during incubation of remoistened soil samples stored for different periods. AB - Periodical measurements for the activities of dehydrogenase, urease, and phosphatase (nuclease) were undertaken during incubation of remoistened alluvial clay and calcareous sandy loam soil samples stored (air-dried) over ten months. All enzyme activities showed fluctuating values within both incubation time and storage period. Dehydrogenase activity tended to increase by storage of samples of both soils. Urease revealed, generally, decreasing values, whilst phosphatase showed no definite trend with the lapse of storage period. PMID- 6285643 TI - [Colony morphology of Clostridium perfringens type A (author's transl)]. AB - At least 4 different colony form of Clostridium perfringens on the surface of yeast extract blood agar after overnight incubation were demonstrated. A: glossy colonies with entire margins, mostly surrounded by narrow zones of complete hemolysis and a much wider zone of incomplete hemolysis (colonies typical for C. perfringens (Fig. 1), B: dwarf colonies, similar to A, but very small (Fig. 2), C:rough colonies slightly raised with lobate margins (Fig. 3-4), D: flat colonies with irregular surface and filamentous margins (Fig. 5). Three strains out of 126 gave rise to glossy and rough colony forms (Fig. 6). PMID- 6285644 TI - [Participation of lipids (sulfocerebrosides) in neuropeptide reception]. PMID- 6285645 TI - [Development in ontogeny of the effect of ACTH fragment 4-7 on rat learning]. AB - Studies have been made of the effect of ACTH fragment 4-7 on learning in rats in early postnatal ontogenesis, as well as of the possibility of preservation of early learning during administration of this peptide to adult animals. It was shown that conditioned reaction of active avoidance practically cannot be formed in normal 13-15-day animals; however, administration of ACTH 4-7 increases the number of animals exhibiting adequate reactions. Weak effect of ACTH 4-7 in 12-15 day animals, in older ones (20-24 days) is changed by a significant stimulating effect. PMID- 6285646 TI - [Antibodies to the hepatitis A virus in the healthy population of Moscow]. AB - Serum specimens collected from 1002 persons in Moscow were tested for the presence of antibodies to hepatitis A virus (anti-HAV antibodies) by solid-phase enzyme immunoassay. The prevalence of these antibodies increased progressively with age from 10% in children aged 5-9 years to over 90% in the age groups of 40 49 years and over, the 50% immunity level being established at the age of 18 years. 79% of infants under 1 year were found to be immune, which was obviously due to the placental transfer of antibodies from mother to child. In a considerable part of seropositive persons over 30 years high or medium antibody titers were detected. These age groups showed a stable proportion of the low, medium and high level of anti-HAV antibodies. The prevalence of such antibodies was not related to sex. The presence of an ample amount of anti-HAV antibodies was determined in all of 18 tested lots of commercial serum immunoglobulin obtained from 3 different manufacturers. PMID- 6285647 TI - [Characteristics of the pKMR plasmids found in Shigella flexneri]. AB - The clinical isolate of Sh. flexneri 1b, resistant to 5 antibiotics and sulfonamides, has been studied by the method of conjugation and found to have a group of transfer-suppressed pKMR-plasmids: pKMR 204-1 (Ap Sm Tc Cm Km Su), pKMR 204-2 (Sm Km Su), pKMR 204-5 (Km Su) and pKMR 204-7 (Sm Tc Cm Km Su), whose molecular weight was 99, 71.2, 73.8 and 59.5 Md respectively. The treatment of the plasmids with restriction endonuclease BamHI has revealed that plasmids pKMR 204-2 and pKMR 204-5 are definitely related to plasmid KMR 204-1, being its deletion mutants. At the same time plasmids pKMR 204-1 and pKMR 204-7 differ in their sensitivity to endonuclease BamHI and stably coexist within the same cell, thus seeming to belong to different compatibility groups. PMID- 6285648 TI - [Relationship of the sensitizing and protective properties of pertussis antigens]. AB - When comparing delayed hypersensitivity (DN) to B. pertussis corpuscular antigens with the agglutinogenic composition of B. pertussis, as well as to its histamine sensitizing, leukocyte-sensitizing, adjuvant and hemagglutinating activity, no correlation was detected between the specific sensitizing properties of these antigens and the serotype and studied nonspecific properties of B. pertussis. The DH level correlated with the protective activity of B. pertussis corpuscular antigens and the ultrasonic fractions of B. pertussis. The close correlation of these two phenomena suggest that DH-inducing and protective B. pertussis antigens are identical, though their action has different manifestations, depending on the method of administration of the antigen preparation. On these grounds a new method for evaluating the immunogenicity of B. pertussis antigens is proposed. This method consists in comparing the sensitizing activity of the antigen under test and that of the national standard. PMID- 6285649 TI - [Mathematical modelling of glycolysis and adenine nucleotide metabolism of human erythrocytes. I. Reaction-kinetic statements, analysis of in vivo state and determination of starting conditions for in vitro experiments]. AB - A mathematical model of energy metabolism of human red cells is presented, which includes besides the glycolytic reactions the adenine nucleotide metabolism. The model is based on the network of chemical reactions, the thermodynamic equilibrium constants of fast reversible reactions and on the kinetic equations for irreversible enzyme reactions. The model consists of a system of 16 differential equations and allows the mathematical evaluation of metabolic levels in the steady state of energy metabolism corresponding to the in vivo state erythrocytes with the kinetic data for the enzymes derived from in vitro experiments. The dependence of the levels of metabolites in the steady state on the activity of some enzymes is analysed to characterize the regulatory properties of the system. The comparison of the steady state levels of the model with experimental data makes it possible to estimate values of some controversial enzyme parameters. Estimates of the kinetic parameters of the following intracellular processes are presented: 1) rate constant of AMP-phosphatase, 2) maximum rate of adenylate deaminase, 3) activity of adenine phosphoribosylpyrophosphate transferase and 4) adenosine transport through the cell membrane. The simulation of the preparatory phase before incubation of erythrocytes indicates, that the model also permits to compute the time course of changes of levels of metabolites. To solve the initial problem the stiff differential equation system is integrated numerically by an efficient program without the application of the quasi-steady-state approximation. PMID- 6285650 TI - [Age related alterations of motor behavior and release of dopaminergic supersensitivity in rats]. AB - Rats aged greater than or equal to 18 months show, aside from clearly diminished motor parameters (exploratory behavior, resting activity, nocturnal activity profile, rotation behavior), after intracerebral dopamine injection a considerably lower apomorphine hypermotility than young animals. The characteristic alteration of activity occurring in young rats during and following chronic administration of haloperidol (1 mg/kg . day, s.c. for 21 days) as an expression of developing dopaminergic supersensitivity does not appear in older animals. Repeated application of amphetamine (2 X 2 mg/kg daily) caused a significant increase in hypermotility. The results are interpreted as the consequence of a age-related reduction of the activity of central-dopaminergic transmission systems and are discussed with regard to possible differences in the realization of agonist- or antagonist-induced supersensitivity. PMID- 6285651 TI - Polyglycolic acid, Dexon subcuticular suture in paediatric orthopaedic surgery. AB - The present investigation was performed to compare the healing of skin incisions following running subcuticular suture using the absorbable material polyglycolic acid (Dexon) (DSS), and our standard procedure using polyamid (Supramid) interrupted percutaneous suture (SPS). In unilateral incisions one half of the wound closure was performed with DSS and the other with SPS. In bilateral incisions one method of closure was used for each side. A total of 100 scars were compared, 50 of each method. Less inflammatory reaction, a broader scar but slightly better cosmetic result and no increase in hypertrophic scar formation was found following Dexon intracuticular suture. Thus we recommend subcuticular suture with absorbable material in children to avoid the inconvenience with the removal of skin sutures. PMID- 6285652 TI - Cholangiocarcinoma in a patient with biliary cysts. AB - A 15-year-old girl with multiple intra- and extrahepatic biliary cysts is presented. At a primary laparotomy a massive dilatation of the choledochal duct was found and a choledochocysto-duodenostomy was made. Initially the patient recovered but 1 1/2 years later a mass reappeared under the right costal arch. At a second laparotomy multiple neoplastic changes were found on the right and left liver lobes. Histologic examination of biopsies from the liver showed the picture of a cholangiocarcinoma. PMID- 6285653 TI - Cytologic examination as an adjunct to laparoscopy and guided biopsy in the diagnosis of hepatic and gallbladder neoplasia. AB - Laparoscopic examination guided biopsy and cytologic examination of samples obtained by aspiration, brush or scrape from the liver or gallbladder were performed on 26 patients suspected of having hepatic or gallbladder neoplasia. Laparoscopic findings were diagnostic of hepatic neoplasia (primary or secondary) in 13 cases and gallbladder carcinoma in 9 cases. The yields of positive diagnoses on biopsy alone, cytology alone and combined biopsy and cytology were 11 (84.6%), 11 (84.6%) and 12 (92.3%), respectively, in cases of hepatic neoplasia and 4 (44.4%), 5 (55.5%) and 5 (55.5%), respectively, in cases of gallbladder carcinoma. The cytologic typing concurred with the histologic typing in six of ten cases (60%) of hepatic neoplasia and all four cases (100%) of gallbladder carcinoma. It is suggested that cytologic examination be used as an adjunct to histologic examination of guided biopsies in the diagnosis of hepatic neoplasia. It is also concluded that it is possible to make a tissue diagnosis in patients with gallbladder carcinoma by obtaining material for cytology and biopsy through laparoscopy. PMID- 6285654 TI - Uterine Mullerian adenosarcoma with psammoma bodies: cytologic, histologic and ultrastructural studies of a case. AB - Epithelial cells associated with psammoma bodies and undifferentiated sarcomatous tumor cells were found in peritoneal washings obtained during laparotomy for removal of metastatic lesions from a uterine stromal sarcoma. Histologic studies of the primary and recurrent tumors showed characteristics of Mullerian adenosarcoma. The cytologic, histologic and ultrastructural features are described. PMID- 6285655 TI - Psammoma bodies in small cell carcinoma of the lung: a case report. AB - Psammoma bodies are ovoid, laminated concretions that, when detected in malignant effusions, suggest the existence of an underlying papillary carcinoma, usually of the thyroid gland, ovary or endometrium. Although psammoma bodies have been reported to arise from a wide variety of papillary tumors, origin in a carcinoma of the lung is uncommon, and origin in small cell carcinoma has not been previously reported. This paper documents an unusual case in which psammoma bodies were detected in a malignant pleural effusion in a patient with small cell carcinoma of the lung. Autopsy demonstrated their site of origin to be visceral pleura of lung and a metastatic nodule on the pleural surface of the diaphragm. PMID- 6285656 TI - Cytopathologic aspects of pulmonary metastasis of malignant fibrous histiocytoma, myxoid variant: fine needle aspiration biopsy of a case. AB - The cytopathologic findings of a fine needle aspiration biopsy of pulmonary metastasis of a malignant fibrous histiocytoma, myxoid variant, are documented. Isolated and irregular loose clusters or bundles of spindle cells and pleomorphic multinucleated giant cells were found. No signet-ring lipoblastlike cells were identified. The cytologic findings were similar to the histology of the primary tumor removed from the patient's leg 18 months previously. PMID- 6285657 TI - Diagnosis of amebic vaginitis from cervicovaginal smears. PMID- 6285659 TI - Investigations of alkaline phosphatase Ca+2-ATPase and Na+, K+-ATPase during beta APN-induced initial bone mineralization inhibition. AB - Tibias of 6-day-old white Leghorn chick embryos treated with beta aminopropionitrile (beta-APN; 0.1 mg/egg/day) for 4 days and injected with 3H proline or 3H-tetracycline on the 11th day were analyzed for incorporation of 3H proline and 3H-tetracycline. The incorporation of 3H-proline was comparable in the controls and beta-APN-treated embryos. However, the incorporation of 3H tetracycline was significantly lower in beta-APN-treated embryos. The bone ash contents were also lower in the latter group. Alkaline phosphatase and Ca+2 ATPase were found to be significantly lower in beta-APN-treated embryonic bones. There was, however, no difference in the activity of Na+, K+-ATPase. The histochemical examination showed the alkaline phosphatase to be present on osteoblasts and matrix vesicle plasma membranes at the periosteal surface. The chick embryonic liver tissue showed no significant differences in the activities of any of the above enzymes. The results suggest that beta-APN-induced inhibition of the bone mineralization may be due to the bone-specific inhibition of alkaline phosphatase and Ca+2-ATPase. PMID- 6285658 TI - The distal nephron in the chick embryo as a target tissue for 1-alpha-25 dihydroxycholecalciferol. AB - A histological and ultrastructural study as well as an autoradiographic analysis after injection of tritiated 1,25-dihydroxycholecalciferol (1,25-DHCC) were conducted on the distal convoluted tubules of chick embryos. Distal tubules in the embryo were shown to have the same spatial distribution as described for the adult kidney; they presented a convoluted portion located in the vicinity of the central intralobular veins and straight portions irradiating from this region towards the periphery. The epithelium in these tubules was well differentiated; its cells had numerous interdigitating folds in their lateral boundaries which were especially numerous at the basal ends. This device greatly increased the membrane surface available for interchange and was interpreted as an expression of active water and/or mineral transport. Nuclear concentration of radioactivity was found 2 h after injection of tritiated 1,25-DHCC in both the pars convoluta and the pars recta of the distal tubules. This concentration could be blocked by the previous administration of large amounts of nonradioactive 1,25-DHCC. These facts were interpreted as indicating that distal convoluted tubules in the chick embryo are functionally differentiated and contain target cells for 1,25-DHCC. PMID- 6285661 TI - Tissue penetration of erythromycin in Waldeyer's ring and bacteriology of secretory otitis media. PMID- 6285660 TI - Ionic mechanisms sustaining activity in ampullar receptors of the frog. AB - Ionic mechanisms sustaining sensory transduction in crista ampullaris sensory cells have been investigated chiefly by replacing the endolymph with solutions with K+ and Ca++ chelators added. The effects of the modified solutions were evaluated by extracellular and intracellular recordings of both and the presynaptic and postsynaptic activity of ampullar receptors. The results strongly suggest that the receptor current in labyrinthine cells is carried exclusively by K+. Moreover evidence are reported indicating that the transducer membrane in ampullar receptors is provided with Ca++ sensitive potassium channels whose opening is depending on free Ca++ released from cupular structures during excitatory deflections. PMID- 6285662 TI - An experimental study on the origin of foam cells in glomerulonephritis. AB - Masugi nephritis induced in cholesterol-fed rabbits is characterized by accumulation of foam cells in glomeruli which vary in number almost in parallel with the natural course of the disease. From electron microscopic appearances these foam cells are apparently derived from migrant blood monocytes and mesangial cells, but not from endothelial cells. Location of monocytic foam cells is extremely variable, though most often observed in the subendothelial space. In addition, monocytes undergo transformation into multinucleated giant cells including Touton type's. Lipid deposits filling the cytoplasm of foam cells seem to be in two forms, free cytoplasmic droplets and membrane-bounded structures partly containing membranous debris or myelin figures. The latter accumulates preferentially in the cytocentrum and is considered to originate from lysosomes. It seems unlikely that glomerular lesions in Masugi nephritis are aggravated by foam cell accumulation. PMID- 6285663 TI - Histological and ultrastructural studies of nephroblastoma in rats induced transplacentally by ethylnitrosourea. AB - Histologic and ultrastructural features of nephroblastomas in Sprague-Dawley rats induced by a single transplacental injection of ethylnitrosourea (ENU) on the 15 16th day of gestation are described and their significance is discussed with respect to the histogenesis of the tumors. Many renal tumors were observed, in addition to predominant occurrence of tumors of the nervous system. Forty-two of 142 offsprings survived over 100 days developed renal tumors, of which 37 rats had nephroblastomas, three rats had cortical adenomas, and one rat had a cystadenoma. The nephroblastomas were found in 35 rats unilaterally and in two rats bilaterally. Most of these tumors had various amounts of hyalinized stroma. In the kidneys with nephroblastomas, isolated, small foci of renal blastema were also occasionally noted. Electron microscopy revealed that nephroblastomas consisted of two types of cells, namely, epithelial and mesenchymal cells. The epithelial cells showed cellular differentiation of varying degree; they are freely lying in the stroma, forming well differentiated tubules or immature tubule-like structures and solid cell nests. Most of the undifferentiated cells observed around the tubular structures and cell nests are demonstrated to have epithelial features. These observations seem to support the concept that nephroblastoma is originated from metanephric blastema. PMID- 6285664 TI - Transmission and scanning electron microscopic studies on the tumors of neuroblastoma group. AB - Ultrastructural study of neuroblastoma group tumors including 7 neuroblastomas, 4 ganglioneuroblastomas, and 2 ganglioneuromas was performed by using both TEM and SEM. Tumor cells showed a wide variation comparable to the developmental stages of nerve cells and were classified into four types according to the neuritic process projections; namely apolar, monopolar, bipolar, and multipolar. Neuroblastoma was composed of small cells of apolar, monopolar, and bipolar types with few multipolar cells. Tumor cells ganglioneuroblastoma and ganglioneuroma were multipolar type. SEM observation demonstrated characteristic varicosities in the elongated neuritis processes, and by TEM examination the dilated portions of these varicosities revealed disruption or disappearance of parallel runnings of the microtubules and microfilaments. TEM examination demonstrated presence of Schwann cells in ganglioneuroblastoma and ganglioneuroma, and satellite cells and perineurial cells in ganglioneuroma. The hitherto undescribed cells with caveolar structure which are thought to be a precursor of these stromal elements were disclosed in ganglioneuroblastoma. In order to evaluate the maturation and prognosis of tumors of this group, stromal differentiation seems to be equally important as ganglionic differentiation because both stromal elements and tumor cells have a common origin of neural crest. PMID- 6285665 TI - Establishment and characterization of two neoplastic cell lines (U-1285 and U 1568) derived from small cell carcinoma of the lung. AB - Two human cell lines have been established in vitro from patients with small cell carcinoma of the lung (SCC). The U-1285 line was derived from the classical small cell type of SCC while the U-1568 originated from a larger cell variant. The cell lines grow as suspension cultures and have been passaged continuously in vitro for 3 years (U-1285) and 2 years (U-1568), respectively. The malignant nature of the lines is suggested by their infinite growth potential, their capacity to form colonies in agarose and their aneuploidy. Both cell lines contain cytoplasmic electron dense particles indistinguishable from classical neurosecretory granules but only in U-1568 has hormone production (human chorion gonadotropin (alpha HCG)) been proven. The U-1568 line produces carcino-embryonic antigen (CEA) while both U-1285 and U-1568 produce alpha-feto-protein (AFP). Chromosome analysis reveal aneuploidy of both lines. Among structural aberrations, involvement of chromosome 14 in U-1285 and chromosomes 1, 7 and 12 in U-1568 is interesting since alterations of these chromosomes have been described previously in other malignant conditions. PMID- 6285667 TI - Male breast at autopsy. AB - In a consecutive and unselected series of 100 male autopsies, the breasts were totally extirpated and histologically investigated. Gynecomasty was found in 55 cases, of which 48 were in healed and seven in still active, intermediate phase. All but two cases were bilateral. No clinical evidence of gynecomasty was established in any case. Thus, gynecomasty seems to be very common in men. In seven cases, severe intraductal epithelial hyperplasia was found, three of pagetoid and four of cribriform type. They belonged to the atypical hyperplastic- early intraductal carcinoma type. All cases were incidentally found and without known clinical relevance. PMID- 6285666 TI - Gynecomasty: histological aspects in a surgical material. AB - In a consecutive and unselected series of 83 patients operated upon for gynecomasty, the histological and related clinical aspects were studied. It was convincingly demonstrated that gynecomasty begins in an active proliferating phase and ends in a fibrous inactive phase. Contrary to previous authors we found it justified that the formation of lobules, indistinguishable from what is seen in the reproduceable age in the female breast, is not necessarily related to the administration of exogenic hormones. In five (6.5%) cases, typical multicentric foci of intraductal epithelial hyperplasia were demonstrated--of the atypical hyperplastic or early intraductal carcinoma type. It was concluded that the clinical significance of such morphological changes must not be overestimated. PMID- 6285668 TI - Inhibition of adrenergic neuroeffector transmission in rabbit pulmonary artery and aorta by adenosine and adenine nucleotides. AB - The effect of adenosine and adenine nucleotides on sympathetic neuroeffector transmission in the rabbit isolated pulmonary artery and aorta was studied. Adenosine (10(-5)-3 x 10(-4)M) decreased the contractile response of pulmonary artery and aorta evoked by electrical-field stimulation. The decrease was reversible. No tachyphylaxis developed. Inhibition of either adenosine deaminase by deoxycoformycin (3.6 x 10(-6)M) or of adenosine transport by dilazep (3 x 10( 6)M) did not alter the inhibitory effect of adenosine on the neurogenic contractions in the pulmonary artery. However, deoxycoformycin plus dilazep markedly enhanced the inhibitory effect of adenosine. The calcium antagonists nifedipine (1.5 x 10(-8)M) and nimodipine (1.3 x 10(-8)M) had no effect on the adenosine-induced inhibition. This was also the case with theophylline (5 x 10( 5)M), atropine (10(-7)M), and the prostaglandin synthetase inhibitors indomethacin (5 x 10(-5)M and suprofen (3 x 10(-5)M). The contractile response of the pulmonary artery elicited by exogenous (-)-noradrenaline (NA; 10(-9)-3 x 10( 4)M) was essentially not altered by adenosine (10(-5)-3 x 10(-4)M). Adenosine (10(-4)M) did not alter the spontaneous 3H-outflow from rabbit aorta preloaded with 3H-(-)-noradrenaline (3H-NA). Adenosine (10(-5)-3 x 10(-4)M), ADP (10(-4)M), ATP (10(-5)M), and inosine (10(-4)M) diminished the overflow of tritium from pulmonary artery and aorta preloaded with 3H-NA. The spontaneous outflow of tritium from aorta preloaded with 3H-NA consisted of 3H-NA (17%), 3H dihydroxyphenylglycol (3H-DOPEG; 30%), 3H-dihydroxymandelic acid (3H-DOMA, 4%), 3H-O-methylated and deaminated metabolites (3H-OMDA; 42%), and 3H normethanephrine (3H-NMN; 2%). Adenosine (10(-5) and 10(-4)M) enhanced 3H-DOPEG and 2H-NMN, decreased 3H-NA, and did not alter 3H-DOMA and 3H-OMDA. The stimulation-evoked overflow of tritium for aorta preloaded with 3H-NA consisted of 3H-NA (31%), 3H-DOPEG (18%), 3H-DOMA (2%), 3H-ONDA (46%), and 3H-NMN (3%). Adenosine (10(-5) and 10(-4)M) enhanced 3H-NA and 3H-DOPEG, decreased 3H-OMDA and did not alter 3H-DOMA and 3H-NMN. Adeosine (10(-6)-10(-4)M) did not alter the accumulation of 3H-NA (10(-8)M) by aorta. It is concluded that adenosine inhibits vascular sympathetic neuroeffector transmission by diminishing the release of transmitter from the nerve terminals. PMID- 6285669 TI - Effect of ethanol and ethanol withdrawal on 3H-muscimol binding and behaviour in the rat: a pilot study. AB - Effect of acute and chronic exposure to and withdrawal from ethanol on 3H muscimol binding in the forebrains and cerebelli and on withdrawal behaviour was investigated in 30 rats. Chronic exposure to and withdrawal from ethanol produced a significant reduction in the number of muscimol binding sites in the cerebelli. A negative correlation was observed between the number of muscimol binding sites in the forebrain and cerebellum in the chronic ethanol and withdrawal groups. Relatively high correlations were found between the number of muscimol binding sites and audiogenic seizures scores during withdrawal from ethanol. Since impaired GABAergic function is associated with increased susceptibility to seizures, there may be a casual relationship between the observed changes in muscimol binding and ethanol-induced audiogenic withdrawal seizures. PMID- 6285670 TI - Dissociation of the effect of captopril on blood pressure and angiotensin converting enzyme in serum and lungs of spontaneously hypertensive rats. AB - Spontaneously hypertensive rats (SHR) of the Okamoto-Aoki strain (n = 40) were treated with captopril (SQ 14,225; D-3-mercapto-2-methylpropanoyl-L-proline) orally, dose 0.2 mg/ml in drinking water. The treatment was initiated early and later during the course of developing hypertension. Continuously treated rats did not develop hypertension. Rats receiving captopril for 12 weeks remained normotensive, whereas withdrawal of the drug resulted in hypertension. Captopril treatment was effective in the rats with established hypertension and decreased the blood pressures to nearly normal values. Serum angiotensin converting enzyme (ACE) activity rose 3-fold in captopril treated rats. ACE in lung plasma membranes increased during captopril treatment, indicating that captopril induced biosynthesis of pulmonary ACE. No qualitative differences were found in the ACE from treated and not treated animals. The dissociation of the antihypertensive effect of captopril and of increased ACE activity in serum and lungs reduce the value of relating blood pressure effects of the drug to measured enzyme activity in the SHR. PMID- 6285671 TI - Cytidine 3',5' Monophosphate (cCMP) is not an Endogenous nucleotide in normal or regenerating rat livers. PMID- 6285672 TI - Interindividual variation in benzo(a)pyrene metabolism and composition of isoenzymes of cytochrome P-450 as revealed by SDS-gel electrophoresis of human liver microsomal fractions. AB - Human liver microsomal fractions from 13 different individuals were characterized with respect to SDS-polyacrylamide gel electrophoretic profiles and regiospecificity in the metabolism of the polyaromatic hydrocarbon benzo(a)pyrene. Pronounced interindividual differences in the composition of microsomal proteins in the molecular weight region of Mr=49,000-60,000 were found. Furthermore, gel electrophoresis combined with staining for peroxidase activity indicated most of the variations among the profiles of microsomal proteins being attributed to interindividual differences in the composition of isoenzymes of cytochrome P-450. One type of human liver microsomal fraction was selectively induced in specimens obtained from patients with known regular drug intake before death and correlated well in molecular weight to the phenobarbital inducible form of cytochrome P-450 in the rabbit. Large variations among the human liver microsomal samples were also seen in the benzo(a)pyrene metabolism. The results indicate the presence of 7-8 different froms of cytochrome P-450 in human liver microsomes and that the interindividual variations seen in drug metabolism may at least in part be explained by interindividual variations in the distribution of these isoenzymes. PMID- 6285673 TI - The action of six lipid perturbing substances on hormone receptor adenylate cyclase complex in turkey erythrocytes and intestinal mucosa cells. AB - Adenylate cyclase and beta-receptor binding in turkey erythrocytes is studied under the influence of dioctyl sodium sulphosuccinate (DSS), sodium dodecyl sulphate (SDS), sodium deoxycholate (DOC), filipin, oxyphenisatine and ethanol. Intact and lysed erythrocytes and erythrocyte membranes were investigated. In general the fluoride stimulated enzyme was most resistant to the action of the substances followed by the ligand binding, the hormone stimulated enzyme being most sensitive to the action of the substances. However, in certain ranges of concentrations, DSS, SDS, filipin and oxyphenisatine could further stimulate the fluoride stimulated enzyme. The adenylate cyclase in intestinal mucosa cells was studied in the presence of DSS, SDS, DOC and oxyphenisatine. The fluoride stimulated enzyme was here less stable than the basal enzyme activity, and two of the substances (SDS and oxyphenisatine) stimulating the erythrocyte enzyme could not stimulate the fluoride stimulated intestinal mucosa cell enzyme. PMID- 6285674 TI - Effects of chlorpromazine on fluid transport across the intestinal mucosa of the rat. AB - The effect of chlorpromazine (CPZ) on net fluid transport in the small intestine of the rat was studied, using a ligated loop model. Given intramuscularly at 5 mg/kg, CPZ induced a 50% increase of net fluid absorption in jejunum, but had no effect in ileum. Similarly, the drug decreased net fluid accumulation induced by dibutyryl-cyclic AMP in jejunum, but did not influence net fluid accumulation in ileum. In contrast, net fluid accumulation induced by cholera toxin was decreased by CPZ in both jejunum and ileum. Inhibition of cyclic AMP (cAMP)-which probably mediates the hypersecretion by the toxin-was also noted. These results suggest that CPZ reverses fluid transport during cholera in two ways: by a cAMP independent enhancement of fluid absorption in jejunum and by inhibition of the toxin-induced cAMP-production in jejunum as well as ileum. Each of these processes may be independently affected by pharmacological substances and subjected to neural and hormonal regulation. PMID- 6285675 TI - Acid soluble and insoluble glycogen in human skeletal muscle. AB - Quantitative biochemical analysis of glycogen was performed with and without perchloric acid pretreatment on biopsy specimens of varying glycogen content, obtained at rest and immediately after exercise after different diets. If analysis was carried out after perchloric acid pretreatment, as is commonly done, lower values were obtained than without such pretreatment. This could be explained by the existence of glycogen in forms of greater and lesser solubility. Evidence for this was obtained by quantitative analysis of glycogen in the perchloric acid extract. The acid soluble form in the extract constituted a greater fraction of total glycogen at supra-normal than at low concentrations. This might also explain why supra-normal levels of glycogen cannot be detected by histochemical methods. The existence of soluble and insoluble glycogen may also be of physiological importance, e.g. by interacting differently with enzymes of glycogen metabolism. PMID- 6285676 TI - Monosynaptic transmission during epileptiform activity induced by penicillin in hippocampal slices in vitro. AB - Synaptic transmission was studied in the CA1 region of transverse hippocampal slices in vitro before and after the addition of the epileptogenic agent sodium benzyl penicillin. The presynaptic fibre volley and the field potential associated with the intracellular EPSP, 'field EPSP', were recorded from the layer of the activated synapses. Addition of penicillin did not change either response. The rising phase of the intracellularly recorded EPSP did not change. However, the peak amplitude and, particularly, the duration of the EPSP both increased. The prolongation of the EPSP may be of importance for the triggering of epileptiform bursts. PMID- 6285677 TI - The level of angiotensin-converting enzyme as indicator of 2-year prognosis in untreated pulmonary sarcoidosis without erythema nodosum. AB - In order to evaluate the prognostic significance of elevated serum angiotensin converting enzyme (SACE) in pulmonary sarcoidosis, the 2-year prognosis was studied in 33 untreated patients without erythema nodosum. Elevated SACE was found in 19 patients among whom total regression or improvement was noted in 42%, against 86% in 14 patients with normal SACE in serial measurements. The groups with and without elevated enzyme levels were not different with respect to sex, age, pulmonary changes, or extrathoracic manifestations. Thus, the findings of elevated SACE in this category of sarcoidosis patients may have some prognostic significance, pointing toward a greater risk of a chronic course. PMID- 6285678 TI - [Beta-lactam antibiotics and the immune system of the body]. PMID- 6285680 TI - [Clinical forms of cancer at onset and differential diagnosis]. PMID- 6285679 TI - [Anticomplement action of carbiphen, amopen and cephalothin in vitro and in vivo]. PMID- 6285682 TI - Mechanism of tumor cell lysis by natural killer cells. PMID- 6285681 TI - Molecular evolution in papova viruses and in bacteriophages. AB - By comparing the DNA sequences of three eukaryotic papova viral genomes, we attempt to show the very close relative phylogeny among the viral species and their host species, and that therefore the viral species appear to have evolved with their hosts. A comparison of the DNA data also reveals that the rate of nucleotide substitutions at the third positions of codons is much faster than at the first and second positions, though the rate varies depending upon the genes. The estimated rates of amino acid substitutions in homologous genes among the three virus species appear to be considerably faster than the rates known for various vertebrate genes. The comparison reveals further that the rate of silent substitutions is faster than that of replacement substitutions. The DNA sequence data on bacteriophages phi X174 and G4 enable us to examine the patterns of nucleotide substitutions in overlapping genes as well as in nonoverlapping genes. It then becomes evident that overlapping genes have a quite different substitutional pattern with respect to the position of nucleotides in codons than do nonoverlapping sequences. In nonoverlapping regions the third positions usually change the fastest among the three codon positions. This pattern does not apply to overlapping genes, which are coded in the same region but with different reading frames. It will be shown that the younger of the two overlapping genes appears to be very tolerant to nucleotide substitutions at any codon position. Further more, the rate of substitution at each nucleotide site appears to be determined by the rate of the corresponding site in the older of the two overlapping genes. PMID- 6285683 TI - Studies on the causes of asthmatic dyspnea. (The cybernetic theory of bronchial asthma). PMID- 6285685 TI - An accurate means of detecting and characterizing abnormal patterns of ventricular activation by phase image analysis. AB - The ability of scintigraphic phase image analysis to characterize patterns of abnormal ventricular activation was investigated. The pattern of phase distribution and sequential phase changes over both right and left ventricular regions of interest were evaluated in 16 patients with normal electrical activation and wall motion and compared with those in 8 patients with an artificial pacemaker and 4 patients with sinus rhythm with the Wolff-Parkinson White syndrome and delta waves. Normally, the site of earliest phase angle was seen at the base of the interventricular septum, with sequential change affecting the body of the septum and the cardiac apex and then spreading laterally to involve the body of both ventricles. The site of earliest phase angle was located at the apex of the right ventricle in seven patients with a right ventricular endocardial pacemaker and on the lateral left ventricular wall in one patient with a left ventricular epicardial pacemaker. In each case the site corresponded exactly to the position of the pacing electrode as seen on posteroanterior and left lateral chest X-ray films, and sequential phase changes spread from the initial focus to affect both ventricles. In each of the patients with the Wolff Parkinson-White syndrome, the site of earliest ventricular phase angle was located, and it corresponded exactly to the site of the bypass tract as determined by endocardial mapping. In this way, four bypass pathways, two posterior left paraseptal, one left lateral and one right lateral, were correctly localized scintigraphically. On the basis of the sequence of mechanical contraction, phase image analysis provides an accurate noninvasive method of detecting abnormal foci of ventricular activation. PMID- 6285684 TI - Hepatitis B viral infection: part I. Clinical features. AB - As a result of early basic biomedical research, the hepatitis B virus has become the best understood cause of viral liver infection in man. The virus has a unique antigenic structure and elicits specific antibody responses. A proper understanding of the serologic events during infection is essential for accurate diagnosis and management. Only 50 percent of infected individuals manifest clinical signs of disease, but 5 to 10 percent develop either a chronic asymptomatic antigen carrier state or chronic hepatitis. Some patients progress to cirrhosis and possibly to primary hepatocellular carcinoma. PMID- 6285686 TI - Task force V: physical interventions and adjunctive therapy. Emergency cardiac care. PMID- 6285688 TI - Inhibition of lectin-binding to saliva-treated hydroxyapatite, to buccal epithelial cells, and to erythrocytes by salivary components. AB - The influence of human saliva on the binding of several lectins to saliva-treated hydroxyapatite surfaces which mimic the teeth, to buccal epithelial cells, and to erythrocytes was determined. Clarified whole saliva inhibited binding of 3H labeled concanavalin A, wheat germ, soy bean, Ulex europaeus, and peanut agglutinins to saliva-treated hydroxyapatite by 75 to 95%, depending on lectin concentration used. Different fractions of saliva-inhibited binding of different lectins. For example, salivary mucins reduced the binding of wheat germ agglutinin to saliva-treated hydroxyapatite, but they did not significantly affect binding of other lectins studied. Components of saliva also inhibited lectin-mediated hemagglutination. Using an indirect enzyme-linked immunoassay, wheat germ and peanut agglutinins were found to bind in a specific manner to human buccal epithelial cells, and this was also greatly reduced by saliva. The data suggest that one of the major functions of salivary and other alimentary secretions may be to reduce the extent of interaction of dietary lectins with mucosal surfaces. PMID- 6285687 TI - Atrophy of the zona fasciculata in the adrenal cortex of thyroparathyroidectomized rats: a quantitative study. AB - Atrophy of the adrenal gland induced by thyroparathyroidectomy (TPX) is attributable primarily to narrowing of the zona fasciculata. But absolute volumes of the adrenal gland and zona fasciculata, not the zona glomerulosa and zona reticularis, were decreased significantly 7 weeks after TPX. Changes consistent with atrophy were also seen in the zona fasciculata by electron microscopy. A slight decrease in volume of cytoplasm in cells of the zona fasciculata was evident 2 weeks after TPX as shown by quantitative stereological techniques. Seven weeks after TPX, there was a significant decrease in average cytoplasmic volume as well as mitochondrial volume and surface density of mitochondrial membranes. A decrease in total volume of the zona fasciculata, as well as volume per cell of mitochondria, correlates directly with decreased production of corticosterone in response to stress in TPX rats. Parathyroidectomy (PX) had no effect on the volume of the adrenal gland, on the volume of the individual cortical zones, or on the volume per cell of cytoplasm or mitochondria; these observations indicate that changes in TPX rats are due primarily to ablation of the thyroid gland. PMID- 6285689 TI - A study of diet, serum lipids, and fecal constituents in spouses. AB - The diet, serum cholesterol, and fecal constituents of 23 healthy spouses have been compared. The pattern of food intake between couples was similar, but of dietary constituents, only dietary fat and fiber intake showed a significant correlation. Of fecal constituents, only fecal fat correlated significantly between couples. PMID- 6285690 TI - Well-differentiated (tubular) carcinoma of the breast. A clinicopathologic study of 145 pure and mixed cases. AB - In this study, 145 well-differentiated (tubular) carcinomas were divided into two groups: 90 pure tubular carcinomas and 55 which were mixtures in that they contained a component of infiltrating duct carcinoma occupying less than half the tumor. Axillary lymph node metastases developed in 29% of women in the mixed group, but occurred in only 6% of the pure group. The prognosis was good in both groups, with five-year-actuarial survival rates in the pure and mixed groups of 100% and 93%, respectively. Residual carcinoma was present in the mastectomy specimen in 28% of the pure group, and in 40% of the mixed carcinomas. In addition, there was a recurrence rate of 50% in patients with pure tubular carcinoma treated by excisional biopsy. These features indicate simple excision of tubular carcinoma is likely to be inadequate therapy and that a mastectomy is warranted. Axillary node dissections should be done when there is a component of infiltrating duct carcinoma because of the increased risk of axillary lymph node metastasis. PMID- 6285691 TI - Spurious rapid infections mononucleosis test results in non-infectious mononucleosis sera. The role of high-titer horse agglutinins. AB - The present report-describes serologic data from five individuals without infectious mononucleosis (IM) who had very high levels of horse red blood cell agglutinins (1:896-1:7168) that were unusual in that they were minimally absorbed by either guinea pig kidney or beef red blood cell suspensions. Agglutination of sheep red blood cells was negative, and Epstein-Barr virus (EBV)-specific serodiagnostic tests revealed evidence for long past primary infections. In one of the five cases, serologic findings suggested a more than usually active EBV carrier state, as evident from persistently high anti-VCA (IgG) levels of 1:1280 and the continual presence of anti-R at 1:40. Sera from all cases predictably interfered with several commercially available IM-specific rapid slide agglutination tests. The incidence of such findings was estimated to be no more than one per one thousand non-IM cases. In three patients adequately studied, the poorly absorbable horse agglutinins persisted in serum samples over periods of five to nine years. The significance of these findings is discussed. PMID- 6285692 TI - The significance of cytoplasmic chondrocyte inclusions in multiple osteochondromatosis, solitary osteochondromas, and chondrodysplasias. AB - Lesions from two patients with multiple osteochondromas and from three patients with solitary osteochondromas were studied. The histologic and ultrastructural features in both conditions were identical. The chondrocyte population in osteochondromas resembled those in normal hyaline cartilage but the cells in osteochondromas exhibited an accumulation of granular and filamentous materials within markedly dilated cisterns of ergastoplasm. These accumulations formed eosinophilic cytoplasmic inclusions which did not stain with Alcian blue, but were PAS positive after diastase digestion. Chondrocytic inclusions of similar or diverse morphology are found in a number of chondrodysplasias. The findings suggest that the solitary osteochondromas, like the lesions of multiple osteochondromatosis, share a common morphologic feature with the chondrodysplasias. PMID- 6285693 TI - Epstein-Barr virus antibody in childhood Hodgkin's disease. AB - Fifteen patients with childhood onset of Hodgkin's disease were studied for prevalence and quantity of Epstein-Barr virus (EBV) antibody to learn about the relationship between infection with EBV and Hodgkin's disease. Findings indicated that, compared with normal child control subjects, prevalence of EBV antibody is not increased in Hodgkin's disease, but the quantity of antibody increases as the duration of Hodgkin's disease increases. It seems that EBV plays no role in the cause of Hodgkin's disease and that production of greater amounts of antibody relates to immunoregulatory defects associated with Hodgkin's disease. PMID- 6285694 TI - Linitis plastica infiltrating the entire gut. AB - A case of extensive spread of linitis plastica, involving the entire gastrointestinal tract, is reported. The tumor probably originated in the stomach. Because the mucosal and serosal layers remained intact, diagnosis was markedly delayed, as is commonly seen in scirrhous carcinoma. The method of spread of the tumor (mainly through the submucosal layer) is unusual, and is probably related to the histological type of carcinoma. PMID- 6285695 TI - Seroepidemiology of infection with hepatitis A virus and hepatitis B virus in the Seychelles. AB - A batch of 417 serum samples obtained from native-born subjects were tested for the presence of hepatitis B surface antigen (HBsAg) and corresponding antibody (anti-HBs), by enzyme-linked immunosorbent assay (ELISA); and antibodies to hepatitis B core antigen (anti-HBc), e-antigen (anti-HBe), and hepatitis A virus (anti-HAV), by radioimmunoassay (RIA). HBsAg was found in only two of the 417 subjects studied. Anti-HBs was detected in 112 samples (26.8%), anti-HBc in 114 (27.3%) and anti-HBe in 31 samples (7.4%). Serologic evidence of a previous or present infection by hepatitis B virus (HBV) was found in 34.5% of the samples studied. Males showed a greater prevalence of anti-HBs and anti-HBc, while anti HBe was more common in females; however, these differences were not significant. With regard to age, a significantly higher prevalence of anti-HBs (p less than 0.05), anti-HBc (p less than 0.025) and anti-HBe (p less than 0.025) was found in the older age groups. Anti-HAV antibodies were present in 90% of the subjects studied, with no variation between the sexes. The anti-HAV rate in the group under 20 years was similar to that found in the older age groups. The total infection rate of hepatitis B virus in the Seychelles is lower than in other tropical areas, HBs antigen/antibody ratio approaching that in temperate areas. Elucidation of the reasons for the low prevalence of hepatitis B virus carriers among the Sevchelles population requires further investigation. PMID- 6285696 TI - Role of viruses in etiology of systemic lupus erythematosus. PMID- 6285697 TI - Endocytosis of lysosomal alpha-galactosidase A by cultured fibroblasts from patients with Fabry disease. AB - The endocytosis of alpha-galactosidase A was studied in cultured fibroblasts from patients with Fabry disease. Alpha-galactosidase A was purified from human placenta by chromatography on concanavalin A-Sepharose, DEAE-cellulose, and N epsilon-aminocaproyl-alpha-D-galactosylamine-Sepharose. Separation of the high uptake form of the enzyme from the low-uptake form was accomplished by chromatography on ECTEOLA-cellulose. With the high-uptake form of the enzyme, the uptake was linear at low concentrations of enzyme and had a Kuptake of 0.01 U/ml of medium that corresponds to a Km of 5.0 x 10(-9) M. At high concentrations of enzyme, it became saturated. The high-uptake form could be converted to the low uptake form by treatment with acid phosphatase. Mannose-6-P strongly inhibited the active uptake of the enzyme. Once taken up into the lysosomes of Fabry disease fibroblasts, alpha-galactosidase A activity was rapidly lost in the first 2 days of incubation at 37 degrees C, but was fairly stable for the next 6 days. The half-life of internalized alpha-galactosidase A activity was calculated to be 4 days. Crosslinking of the enzyme with hexamethylene diisocyanate did not increase the intracellular stability of alpha-galactosidase A activity. PMID- 6285698 TI - Occupational and environmental health problems of the developing oil shale industry: a review. AB - The American oil shale industry is on the threshold of commercial industrial development. Potential occupational hazards include shalosis or oil shale pneumoconiosis, dermatoses, cancer of the skin, lung, and possibly other sites, and accidents. Air, water, and solid waste pollution problems are complicated by the aridity of the Green River oil shale formation located in Utah, Colorado, and Wyoming. The region currently lacks the schools, health facilities, community services, and skilled labor required for large-scale development. The oil shale industry faces an opportunity and a challenge of prudently assessing and controlling exposures and contributing to the social development of the region. PMID- 6285699 TI - Acyclovir treatment of experimental genital herpes simplex virus infections. AB - Genital herpes simplex virus type 2 (HSV-2) infections in mice and guinea pigs were used to determine the effectiveness of acyclovir administered topically, orally, or parenterally. Topical treatment with 1 percent or 5 percent acyclovir begun six or 24 hours and 5 percent acyclovir begun 48 or 72 hours after intravaginal inoculation of mice with HSV-2 significantly inhibited viral replication in the genital tract. Oral administration of acyclovir resulted in a significant reduction in vaginal virus titers when therapy was begun as late as 72 hours after infection. Topical treatment with 5 percent acyclovir initiated 24, 48, or 96 hours after intravaginal inoculation of guinea pigs with HSV-2 significantly altered lesion severity and virus titers but not vaginal HSV titers. Oral acyclovir therapy begun 24 or 48 hours after infection also significantly reduced the mean lesion score and prevented their development in some animals. Lesion and vaginal virus titers were only partially reduced. Intramuscular administration of acyclovir begun on day 2 or day 4 of infection significantly reduced the mean lesion scores but failed to alter lesion or vaginal virus titers. PMID- 6285700 TI - Acyclovir in the treatment of simian varicella virus infection of the African green monkey. AB - Acyclovir (9-(2-hydroxyethoxymethyl)guanine) administered as an intramuscular formulation twice daily at a dosage of 100 mg/kg per day prevented the development of disease in African green monkeys inoculated with simian varicella virus. Viremia, appearance of a vesicular rash, and elevations of serum transaminases, each indicative of infection, were suppressed by acyclovir treatment. Plasma concentrations of acyclovir were measured and showed rising levels after repeated intramuscular injection with a prolonged period of absorption of acyclovir into the plasma circulation. Investigation of antiviral efficacy after either intramuscular or intravenous acyclovir treatment showed both routes of administration to be effective in inhibiting simian varicella virus infection at the 100 mg/kg per day level. However, intravenous acyclovir 50 mg/kg twice daily did result in elevated values of blood urea nitrogen, creatinine, and serum transaminases. PMID- 6285701 TI - Acyclovir in mouse cytomegalovirus infections. AB - A virus-specified thymidine kinase appears to be a general requirement for herpes virus susceptibility to the antiviral effect of acyclovir. Surprisingly, mouse cytomegalovirus (MCMV), which does not encode for a thymidine kinase, is exquisitely sensitive to the drug both in vitro and in vivo. The drug is active against the virus in the absence of a cellular thymidine kinase and the antiviral activity is not diminished in the presence of excess thymidine or a variety of nucleosides and deoxynucleosides. Thus, a thymidine phosphorylation pathway is not required for the drug's activation of this infection. The enzyme system responsible for phosphorylation of the drug has not been identified. Mouse cytomegalovirus mutants resistant to the drug have been isolated, indicating that the antiMCMV effect results from selective inhibition of viral replication rather than indirectly through toxicity to the host cell. Eight resistant mutants appear to be in the same complementation group and seven of the mutants demonstrate coresistance to phosphonoacetic acid, a marker for the DNA polymerase locus of herpes viruses. The evidence to date indicates that the MCMV DNA polymerases is the final site of action of the drug. Investigations of the antiMCMV activity of acyclovir should provide insights into the antiviral effects of this drug and other nucleoside analogs in other herpes virus infections in which the virus does not code for a thymidine kinase (for example, human cytomegalovirus and Epstein Barr virus). PMID- 6285702 TI - Effect of acyclovir treatment on acute and chronic murine cytomegalovirus infection. AB - Murine cytomegalovirus (MCMV) is inhibited in vitro by 1 to 2 microM acyclovir. Therapy of a systemic MCMV infection in weanling mice with acyclovir was only minimally effective when drug was administered intraperitoneally, while oral administration by addition of acyclovir to the drinking water was highly efficacious in mice with disseminated MCMV. Effective therapy was characterized by reduction of virus titers in lung, liver, spleen, and kidney. In mice chronically infected with MCMV, treatment for 30 days with oral acyclovir eliminated or reduced virus titers in all target organs except the salivary gland. Therapeutic efficacy in this model infection using oral administration of acyclovir could be correlated with the achievement of acyclovir levels in the plasma of experimental animals two to 10 times greater than the mean inhibitory concentration for MCMV in vitro throughout treatment. The lack of efficacy observed when drug was administered intraperitoneally was associated with acyclovir levels exceeding 1 microM for one to three hours after each dose. PMID- 6285704 TI - Effect of acyclovir on the deoxyribonucleoside triphosphate pool levels in Vero cells infected with herpes simplex virus type 1. AB - The effect of acyclovir on the deoxyribonucleoside triphosphate pools of Vero cells infected with herpes simplex virus type 1 was examined. Deoxyguanosine triphosphate and deoxyadenosine triphosphate pool levels in infected cells treated with acyclovir increased dramatically compared with pool levels in untreated infected cels. The increases were due, at least in part, to inhibition of viral DNA polymerase activity which resulted in reduced utilization of the deoxyribonucleoside triphosphates. Differences of as much as 26 times were detected in the sensitivity of herpes simplex virus type 1 to inhibition by acyclovir with different Vero cell cultures. These results were due to differences in acyclovir triphosphate levels, not to differences in deoxyguanosine triphosphate levels. PMID- 6285703 TI - Treatment of experimental latent herpes simplex virus infections with acyclovir and other antiviral compounds. AB - Acyclovir can prevent the establishment of latent herpes simplex virus (HSV) in sensory ganglia by early treatment of the primary infection in experimental animals. A delayed treatment may reduce the number of neurons that eventually become latently infected. Acyclovir can prevent virus reactivation in explant cultures of latently infected ganglia, experimentally induced recurrences of latently infected animals, and immunosuppressed patients. This effect is dependent on the continuous presence of the drug in the explant culture medium or organism. The treatment of recurrent lesions with acyclovir reduces the severity of the episode and may also prevent reinfection of ganglia, if this should be common in the maintenance of latency. It might be possible to eradicate a latent HSV, if conditions were created in which drugs such as acyclovir, activated by the virus-induced thymidine kinase, could interact with the enzyme before the assembly of mature virions. PMID- 6285707 TI - Evaluation of the antiherpetic activity of acyclovir in rabbits. AB - Acyclovir (3 percent ointment) used topically one to five times a day on acute ocular herpes simplex virus (HSV) infection gave beneficial results as measured by a reduction in corneal involvement, conjunctivitis, iritis, and corneal clouding. Early intensive acyclovir topical treatment every two hours beginning 24 hours after inoculation prevented all but conjunctivitis. Topical treatments did not prevent the establishment of latent HSV infection. Effects of topical acyclovir treatment on rabbit eyes infected with parent McKrae, idoxuridine resistant, and vidarabine-resistant strains of HSV-1 were studied. Topical acyclovir therapy given four times a day was significantly more effective than idoxuridine and vidarabine in suppressing acute herpetic ocular disease induced by either sensitive or drug-resistant strains of HSV-1. Virus isolation from neural tissues indicated that none of the therapy prevented viral infection in the nervous system. Intravenous acyclovir used twice a day (50 mg/kg) on rabbits with latent HSV infection appeared to suppress HSV in the nervous system but did not eradicate established latent HSV infection. PMID- 6285706 TI - Effect of acyclovir, bromovinyldeoxyuridine, vidarabine, and L-lysine on latent ganglionic herpes simplex virus in vitro. AB - Vidarabine and L-lysine did not prevent the in vitro reactivation of latent herpes simplex virus or block the further multiplication of reactivated latent virus. Acyclovir and bromovinyldeoxyuridine both blocked the reaction and the multiplication of reactivated latent virus, and transiently suppressed but did not eliminate latent virus from the sensory ganglia. PMID- 6285705 TI - Effect of acyclovir on genital infection with herpes simplex virus types 1 and 2 in the guinea pig. AB - The pathogenesis of genital infection with three different strains of herpes simplex virus type 1 (HSV-1) and three strains of herpes simplex virus type 2 (HSV-2) was compared in the guinea pig. Strain differences in severity of clinical disease and mortality were noted. HSV-1 strains generally produced milder disease than HSV-2. Both HSV-1 and HSV-2 infections resulted in acute and chronic changes in the cervix. Virus recovery during latent infection was more frequently obtained from the spinal cord in HSV-1-infected animals and from lumbosacral ganglia in HSV-2-infected animals. Systemic treatment with acyclovir, after the onset of clinical disease, had minimal, if any, effect on genital infection with HSV-1 (NYU-78), but similar treatment of HSV-2 (WT-186) infection resulted in decreased lesion scores, paralysis, and mortality during acute infection. A reduction in virus isolations from lumbosacral ganglia was noted during both acute and latent infection with HSV-2 (WT-186) in the acyclovir treated groups. PMID- 6285708 TI - Preventive and curative effects of acyclovir on central nervous system infections in hamsters inoculated with herpes simplex virus. AB - Hamsters are very susceptible to infections with herpes simplex virus. Inoculation of the skin of the lower back region leads to an ascending involvement of the central nervous system with fatal outcome. This model was used for testing the activity of antiviral substances in herpes infections of the nervous system. Acyclovir given in optimal dosages prevented infection of the central nervous system completely even when therapy was started 48 hours after inoculation. Therapy with acyclovir proved to be successful in animals showing paresis after infection even when the drug was given in lower than optimal dosages. PMID- 6285709 TI - Acyclovir tolerance in humans. AB - Acyclovir tolerance has been explored in a broad range of human populations and dosage regimens with intravenous, topical, and oral formulations. Phase I pharmacokinetic/tolerance studies assured safety in special populations at unique risk of complicated herpes infections who were simultaneously at increased risk of toxicity to anti-DNA chemotherapeutic agents. Further safety evaluations accompanied placebo-controlled Phase II studies in infected patients who represent future users of acyclovir. These studies confirm acyclovir as the safest antiherpes agent to be explored in clinical studies to date. PMID- 6285710 TI - Perspectives on interactions of acyclovir with Epstein-Barr and other herpes viruses. AB - Acyclovir [9-(2-hydroxyethoxymethyl)guanine] inhibits Epstein-Barr virus (EBV) replication in lymphoblastoid cells at concentrations nontoxic to cellular growth. The mode of action of the drug against EBV differs from the mechanism described in herpes simplex virus systems. Due to the absence of virus-specified thymidine kinase, the drug is poorly phosphorylated in EBV-infected cells. The extent of monophosphorylation is similar both in mock-infected and EBV-infected cells. Despite weak phosphorylation of the drug, the replication of linear EBV DNA is inhibited due to exquisite sensitivity of the viral DNA polymerase. Activation of acyclovir does not require phosphorylation by virus-specified thymidine kinase, inhibition of different herpes-group viruses depends on three variable factors: degree of phosphorylation, cellular metabolism of the drug, and degree of sensitivity of the viral polymerase. Interaction of acyclovir triphosphate with EBV DNA polymerase is reversible. Cells infected with EBV and treated with acyclovir resume virus replication following removal of the drug even after long exposure. Acyclovir inhibits replication of linear genomes and stops production of virus, but has no effect on latent cellular infection. These results lead us to predict that acyclovir will suppress, but not cure, EBV infection. PMID- 6285711 TI - Renal function after acyclovir intravenous injection. AB - Plasma urea or creatinine was noted to be raised in 58 of 354 patients treated with intravenous acyclovir. This occurred after intravenous bolus injection of high dosages of acyclovir but the risk was considerably reduced by slow intravenous infusion of the same dosage over one hour, with adequate hydration of the patient and adjustment of dosage in patients with preexisting impaired renal function. Animal studies indicate that the impairment of renal function associated with high bolus injections of acyclovir is due to crystal formation in the renal tubules and/or the collecting ducts, and that the crystals are removed after cessation of treatment. The special problems involved in the treatment of patients with herpes encephalitis necessitating limited fluid intake and the possible interaction with other nephrotoxic drugs are discussed. PMID- 6285712 TI - Use of acyclovir in premature and term neonates. AB - Nine infants with symptomatic infections caused by herpes simplex virus or cytomegalovirus were treated with acyclovir. At the onset of therapy, the infants ranged in weight from 880 to 4550 gm. Five were premature. Acyclovir was administered intravenously in a dosage of 5 to 15 mg/kg every eight hours for five to 10 days. The peak serum acyclovir levels ranged from 20 to 163 microM and the trough levels ranged from 1 to 129 microM. The variation in peak serum acyclovir levels in different infants receiving the same dosage on a weight basis was large but correlated with the expected renal maturity of the individual infant. Hematologic values improved during therapy. No renal toxicity was noted. All of the infants survived, including the five with herpes simplex infections. PMID- 6285713 TI - Neonatal acyclovir pharmacokinetics in patients with herpes virus infections. AB - A preliminary analysis is presented of the pharmacokinetics of acyclovir in neonatal patients with herpes simplex virus infections. Mean peak acyclovir levels (microM +/- SD) at 5, 10, and 15 mg/kg per dose were 30.0 +/- 9.9, 61.2 +/ 18.3, and 86.1 +/- 23.5, with corresponding mean trough levels (microM +/- SD) of 5.3 +/- 3.4, 10.1 +/- 8.4, and 13.8 +/- 11.1, respectively. The mean half-life (t 1/2 beta) of acyclovir was 3.78 +/- 1.21 hours. The mean percent urinary recovery of acyclovir (+/- SD) at each dosage level was similar, with an overall mean recovery of 65 percent. The mean acyclovir concentration in urine did not exceed the solubility of acyclovir in bladder urine (1,300 micrograms/ml). Generally, neonatal acyclovir pharmacokinetics was consistent with previous reports from studies of adults. PMID- 6285714 TI - Acyclovir treatment of herpes simplex virus infections in immunocompromised humans. An overview. AB - As reflected in the preclinical and early clinical data, acyclovir seems destined to have a very useful role in the treatment and/or prophylaxis of herpes virus (HSV) infections in immunosuppressed humans. If the preclinical predictions can be extrapolated to varicella-zoster (V-Z) infections, acyclovir could well also play a meaningful role in therapy of V-Z infections in the immunosuppressed host; however, this conjecture awaits proof of controlled studies in humans. Clearly the usefulness of acyclovir for treatment of V-Z infections should be compared with that of adenine arabinoside (ara-A) to put into proper perspective the relative efficacies of the two drugs for future therapeutic regimens. The need for comparative studies is most important at this early stage of antiviral drug development, to avoid ethical problems that will cloud the knowledge needed to move forward in a positive way. In any event, the development of acyclovir, with its targeted approach, represents a real fundamental advance in antiviral drug development. Together with the development and deployment of ara-A, it should provide the needed impetus for a surge in the creation of new antiviral compounds for tomorrow. PMID- 6285715 TI - Acyclovir in immunocompromised patients with cytomegalovirus disease. A controlled trial at one institution. PMID- 6285716 TI - Treatment of cytomegalovirus pneumonia with high-dose acyclovir. AB - Cytomegalovirus pneumonia is a serious complication of marrow transplantation, with a 90 percent fatality rate. Acyclovir, a new antiviral agent with variable in vitro activity against cytomegalovirus, was administered to eight marrow transplant patients with biopsy-proven cytomegalovirus pneumonia; one patient survived. Doses were between 400 and 1200 mg/m2 and peak plasma levels between 47 and 316 microM were attained. Possible marrow toxicity occurred in three patients, and mild neurotoxicity occurred in one. High-dose acyclovir had mild toxicity but was not effective as treatment for cytomegalovirus pneumonia after marrow transplantation. PMID- 6285717 TI - In vitro and in vivo responses of cytomegalovirus to acyclovir. AB - Laboratory strains and fresh isolates of human cytomegalovirus (HCMV) were tested for sensitivity to acyclovir. Fifty percent inhibition was achieved at about 10 micrograms/ml, but 25 to 100 micrograms/ml was necessary for 90 percent or greater inhibition. Acyclovir was administered intravenously to four patients with congenital cytomegalovirus (CMV) infection. There was only a temporary diminution of virus titer in the urine and no obvious clinical improvement. One child experienced hematuria. Pharmacokinetic studies showed a large volume of distribution and slow excretion of acyclovir with a half-life of approximately three hours. PMID- 6285718 TI - Treatment of life-threatening Epstein-Barr virus infection with acyclovir. AB - Two pediatric patients with life-threatening Epstein-Barr virus infections were studied immunologically and treated with acyclovir [9-(2-hydroxyethoxymethyl) guanine]. The patient with chronic active Epstein-Barr virus infection who experienced massive hepatosplenomegaly, pancytopenia, and failure to thrive demonstrated abnormalities of T and B lymphocytes. A second patient, with the X linked lymphoproliferative syndrome, experienced a rapidly fatal course of acute Epstein-Barr virus infection which typifies this yet undefined immunodeficiency to Epstein-Barr virus. In each case, objective evidence for clinical improvement or antiviral effect of acyclovir treatment was not apparent. Abnormally productive Epstein-Barr virus infections did not appear to play a major role in the clinical syndromes observed. Current studies are focused on treatment of immunologically normal patients with early complicated Epstein-Barr virus infection. PMID- 6285719 TI - Treatment of herpes virus infections in immunocompromised patients with acyclovir by continuous intravenous infusion. AB - Sixteen immunocompromised patients with herpes virus infections were treated for three to five days with continuously administered intravenous acyclovir. Patients received initial acyclovir infusions over 5 minutes in dosages ranging from 1.5 to 5.0 mg/kg followed by continuously infused acyclovir at 7.2, 14.4, 21.6, 28.8, 36.0, or 43.2 mg/kg per day. The mean serum plateau levels of acyclovir determined by radioimmunoassay ranged from 4.1 microM for the 7.2 mg/kg per day dosage to 36.6 microM for the 43.2 mg/kg per day dose. A mean of 75 percent of acyclovir administered was recovered in the urine of patients treated. Eleven of 13 patients with varicella-zoster virus (VZV) infections had no new vesicle formation after three days of acyclovir treatment and all patients ceased to have new vesicles after five days of therapy. For the nine patients from whom complete viral cultures were available, six ceased to shed virus at three days, and viral shedding ceased by five days in all patients treated with acyclovir. No clinical or laboratory adverse reactions were associated with acyclovir therapy. These data suggest that acyclovir given by continuous intravenous infusion may be useful in the treatment of herpes virus infections in immunocompromised patients. PMID- 6285721 TI - Topical 5 percent acyclovir in polyethylene glycol for herpes simplex labialis. Antiviral effect without clinical benefit. AB - A double-blind, placebo-controlled trial of topical 5 percent acyclovir in polyethylene glycol was conducted among 208 patients with herpes simplex labialis. Reduced excretion of virus from lesions was seen in the subgroup of patients who entered the study within eight hours of lesion onset, but no differences were noted in the patients who began treatment nine to 25 hours after onset. No clinical benefit from treatment with acyclovir was observed. PMID- 6285722 TI - Biochemical and genetic analysis of acyclovir-resistant mutants of herpes simplex virus type 1. PMID- 6285720 TI - Chemotherapy of herpetic keratitis induced by acyclovir-resistant strains of herpes simplex virus type 1. AB - The severity of herpetic keratitis induced by 9-(2-hydroxyethoxymethyl) guanine resistant strains of herpes simplex virus was significantly reduced by cotherapy with 9-beta-D-arabinofuranosyladenine (ara-A) and 2-deoxycoformycin. Therapy with 5-trifluoromethyl-2'-deoxyuridine (F3TdR) significantly reduced the severity of keratitis induced by an acyclovir-resistant strain with a defective DNA polymerase. Therapy with 3 percent acyclovir ointment slightly reduced the number of herpetic lesions produced by either deoxypyrimidine kinase or DNA polymerase defective viruses, despite these viruses being 100 to 1000 times more resistant to acyclovir than the wildtype strain. Therapy with 3 percent ara-A ointment alone significantly reduced the severity of lesions produced by the wildtype herpes strain. Therapy with ara-A alone did not reduce the severity of disease induced by any of the acyclovir-resistant mutants. The sensitivity of the wildtype and mutant viruses to nucleoside analogs was confirmed by yield reduction assays conducted with Vero cells. These studies indicate that cotherapy with ara-A and an adenosine deaminase inhibitor was a reasonable alternative therapy for keratitis due to mutants resistant to therapy with nucleoside analogs which require the virus-specified deoxypyrimidine kinase or DNA polymerase, while ara-A alone was not an effective alternative. PMID- 6285723 TI - Genetic mechanisms of resistance to acyclovir in herpes simplex virus. PMID- 6285724 TI - Isolation and characterization of acyclovir-resistant strains of herpes simplex virus. AB - A number of clinical studies have documented herpes simplex infections which appear to be resistant to nucleoside analogs; these include idoxuridine [1,2] and acyclovir [3]. Few, if any of the viruses isolated from such patients have yet been thoroughly characterized. We have isolated a number of acyclovir-resistant mutants by selection for resistance in tissue culture. The study of the biochemical and biological properties of these mutants has given some insight into the likely nature of resistant clinical strains. We have devised a number of simple tests to allow classification of laboratory mutants. We also draw attention to some of the difficulties the clinical virologist may encounter when analyzing putative resistant virus isolated from treated patients. PMID- 6285725 TI - Disease and latency characteristics of clinical herpes virus isolated after acyclovir therapy. AB - Herpes simplex virus (HSV) type 1 from a bone marrow transplant recipient and HSV type 2 from a patient with genital herpes infection were examined for sensitivity to acyclovir after both patients received therapy with the drug. A 38- and 83 fold shift in sensitivity was detected in association with a marked decrease in viral thymidine kinase activity in isolates from both patients. The resistant HSV 1 isolate was approximately 900 times less neurovirulent to Balb/C mice but had similar cutaneous virulence in hairless mice compared with the patient's sensitive strain. In contrast, there was no difference in pathogenicity between the sensitive and resistant HSV-2 isolates. Latency was detected in the trigeminal ganglia of mice after snout inoculation with both the sensitive and resistant HSV-1 isolates. The ganglion isolate from the resistant HSV-inoculated mouse was found to be sensitive to acyclovir, implying a selection for or reversion of the sensitive phenotype. No trigeminal ganglion latency was detected after inoculation with either HSV-2 isolate. Resistance to acyclovir can arise during therapy as a result of diminished viral thymidine kinase activity but does not appear to be associated with increased virulence. PMID- 6285726 TI - Spectrum of sensitivity of acyclovir of herpes simplex virus clinical isolates. AB - HSV-1 and HSV-2 clinical isolates were tested for their in vitro sensitivity to acyclovir. The median ID50 for 32 genital HSV-2 isolates was 0.215 micrograms/ml and this was not significantly higher than a median value of 0.125 micrograms/ml for 28 oral HSV-1 isolates (p = 0.08). A wider range of ID50s was observed with the HSV-2 isolates compared with the HSV-1 isolates. The concentration of drug required to inhibit virus replication by 90 and 99 percent was approximately 10- and 100-fold greater than that producing 50 percent inhibition. PMID- 6285727 TI - Sensitivity of herpes virus isolates from acyclovir clinical trials. AB - Clinical isolates of herpes simplex virus and varicella zoster virus have been examined for changes in drug sensitivity after treatment of patients with acyclovir. Some preliminary sensitivity results determined by plaque reduction assays in Vero cells for herpes simplex virus and MRC-5 cells for varicella zoster virus are presented. Where both pretreatment and posttreatment virus isolates were available, no significant changes in sensitivity to acyclovir were observed. Similarly, isolates taken after treatment was begun, without a pretreatment sample, gave IC50 values no greater than expected when compared with other pretreatment isolates. PMID- 6285728 TI - Selection and preliminary characterization of acyclovir-resistant mutants of varicella zoster virus. AB - A series of acyclovir-resistant mutants of varicella zoster virus (VZV) were selected in vitro by serial passage of VZV-infected human fibroblasts in increasing drug concentrations, or by continuous exposure of cultures infected at high multiplicity to 100 microM acyclovir. The in vitro susceptibility of these mutants to several antiherpetic agents was measured by the plaque-reduction assay. The capacity of extracts of cells infected with these mutants to phosphorylate acyclovir was examined and compared with that of their acyclovir sensitive parent strains. Based on these studies, VZV could be shown to acquire resistance to acyclovir through diminished acyclovir phosphorylation. This was presumable due to loss of viral specific thymidine kinase (TK) function. Two acyclovir-resistant mutants remained TK competent but demonstrated phenotypic changes in sensitivity to antiviral agents known to act at the herpes simplex virus (HSV)-specific DNA polymerase level. These results suggest that the resistance of VZV to acyclovir results from qualitative or quantitative alterations in the virus-specified TK or DNA polymerase. PMID- 6285730 TI - Acyclovir chemistry and spectrum of activity. AB - Most antiviral agents with activity against the herpes group of viruses belong to a class of compounds known as nucleosides. Recently, an acyclic analog of a natural nucleoside has been found to have high activity against the herpes virus. This compound, called generically acyclovir, is 9-(hydroxymethyl)guanine. Its spectrum of antiviral activity is described. PMID- 6285729 TI - Drug-resistant herpes simplex virus in vitro and after acyclovir treatment in an immunocompromised patient. AB - An acyclovir-resistant herpes simplex virus (HSV) has been isolated from an immunocompromised patient during treatment for severe orofacial HSV infections with acyclovir. The resistant virus is deficient in expression of the HSV thymidine kinase (TK). Emergence of acyclovir resistance during clinical use appears to parallel the in vitro observations of selection for TK-deficient, acyclovir-resistant viruses following serial passage in the presence of the drug. The clinical importance of drug-resistant HSV in unclear, and further investigations are required to determine whether it will be an impediment to successful therapy of HSV infections. PMID- 6285731 TI - Different in vitro effects of dual combinations of anti-herpes simplex virus compounds. AB - Five promising antivirals have been tested individually and in pairs on herpes simplex virus (HSV) types 1 (Strain F) and 2 (Strain G) in Vero cells. These are: 9-(2-hydroxyethoxymethyl)guanine (acyclovir, ACV), 9-beta-D arabinofuranosyladenine (ara-A), 5-trifluorothymidine (TFT), E-5-(2-bromovinyl) 2'-deoxyuridine (BVDU), and phosphonoformate (PFA). Various types of interactions depending on virus type were observed: synergistic [ara-A/TFT; ara-A/BVDU (G); BVDU/PFA (F)]; additive [ACV/ara-A; ACV/TFT; ACV/BVDU; ACV/PFA (G); BVDU/TFT; PFA/ara-A; PFA/TFT]; and sub-additive [ACV/PFA (F); ara-A/BVDU (F) and BVDU/PFA (G)]. Neither antagonism nor interference was noted for any combinations. PMID- 6285732 TI - Effectiveness of acycloguanosine and trifluorothymidine as inhibitors of cytomegalovirus infection in vitro. AB - The effectiveness of acycloguanosine (acyclovir) and trifluorothymidine (TFT) as inhibitors of cytomegalovirus (CMV) infection was tested using a plaque-reduction assay. Two laboratory strains and seven clinical isolates were employed. The results indicated that both acyclovir and TFT were ineffective in inhibiting viral cytopathic effect (CPE) below the concentration of 50 microM. At concentrations of 100 microM or more, greater than a 50 percent inhibition was achieved by both acyclovir and TFT on all clinical isolates. Although complete inhibition of plaque formation was seldom achieved even at a drug concentration as high as 200 microM, a reduction of plaque size was consistently observed at drug concentrations above 100 microM. When a low dose of virus was used for infection, ED50 could be achieved at an acyclovir concentration of 50 microM. Laboratory strains of CMV were found to be more resistant than the clinical isolates to inhibition of acyclovir. PMID- 6285733 TI - Effect of acyclovir combined with other antiherpetic agents on varicella zoster virus in vitro. AB - The sensitivity of varicella zoster virus (VZV) strain Ellen to acyclovir in combination with other antiherpetic agents in vitro has been examined by the plaque-reduction and infectious center assay methods. (E)-5-(2-bromovinyl)-2' deoxyuridine (BVDU), trifluorothymidine (TFT), or 5-iodo-2'-deoxyuridine (IdUrd) produced, in general, additive antiviral activity when examined by either assay method. Acyclovir in combination with phosphonoacetate (PAA) resulted in additive effects as measured by the plaque-reduction assay. The combination of acyclovir with 9-beta-D-arabinofuranosyladenine (ara-A) gave generally additive or occasionally synergistic antiviral activity when tested in MRC-5 or WI-38 cells respectively, regardless of assay method. PMID- 6285734 TI - Activation and antiviral effect of acyclovir in cells infected with a varicella like simian virus. AB - Acyclovir inhibited the replication of a varicella-like simian virus (DHV-1) in cell culture (Vero cells) with an ED50 of 38 +/- 2 microM. The activation of acyclovir in this cell culture system was compared with that in the cell system with human varicella zoster virus (VZV). Extracts of cells infected with DHV-1 catalyzed the phosphorylation of acyclovir. The phosphorylation was inhibited by dThd, suggesting the catalyst was a dThd kinase. Electrophoresis of cytosol fractions on polyacrylamide gels corroborated the existence of a virus-associated dThd kinase. This enzyme copurified with an acyclovir-phosphorylating activity. The enzyme catalyzed the phosphorylation of acyclovir at a greater relative rate than that with the VZV enzyme, but with a higher apparent Km value for acyclovir. The relative efficiencies for the two enzymes with acyclovir were similar. Anabolic studies with cells infected with DHV-1 and incubated with [14C]acyclovir indicated that triphosphate of acyclovir did accumulate. The results indicate that acyclovir is activated in cells infected with DHV-1 in a manner similar to that in cells infected with VZV. PMID- 6285735 TI - Acyclovir inhibition of viral DNA chain elongation in herpes simplex virus infected cells. AB - The effect of acyclovir on DNA synthesized in HSV-1-infected cells was examined. Viral DNA synthesis was significantly reduced in the presence of 10 microM acyclovir monitored by cRNA-DNA hybridization and isopycnic centrifugation in cesium chloride. DNA synthesized in infected cells in the presence of acyclovir appeared to be chain-terminated when evaluated by rate zonal centrifugation in alkaline sucrose. Cellular DNA synthesis in actively replicating uninfected cells in the presence of acyclovir was evaluated by isopycnic centrifugation. Cellular DNA synthesis appeared unaffected at concentrations of acyclovir up to 100 microM. PMID- 6285736 TI - Mechanism of action and selectivity of acyclovir. AB - Acyclovir, an acrylic purine nucleoside analog, is a highly potent inhibitor of herpes simplex virus (HSV), types 1 and 2, and varicella zoster virus, and has extremely low toxicity for the normal host cells. This selectivity is due to the ability of these viruses to code for a viral thymidine kinase capable of phosphorylating acyclovir to a monophosphate; this capability is essentially absent in uninfected cells. The acyclovir monophosphate (acyclo-GMP) is subsequently converted to acyclovir triphosphate (acyclo-GTP) by cellular enzymes. Acyclo-GTP persists in HSV-infected cells for many hours after acyclovir is removed from the medium. The amounts of acyclo-GTP formed in HSV-infected cells are 40 to 100 times greater than in uninfected Vero cells. Acyclo-GTP acts as a more potent inhibitor of the viral DNA polymerases than of the cellular polymerases. The DNA polymerases of HSV-1 and HSV-2 also use acyclo-GTP as a substrate and incorporate acyclo-GMP into the DNA primer-template to a much greater extent than do the cellular enzymes. The viral DNA polymerase binds strongly to the acyclo-GMP-terminated template, and in thereby inactivated. PMID- 6285737 TI - Effect of acyclovir on herpes simplex virus replication in a persistently infected human lymphoblastoid cell line (P3HR-1). PMID- 6285738 TI - Replication of Epstein-Barr virus DNA in lymphoblastoid cells treated for extended periods with acyclovir. PMID- 6285739 TI - Effect of 9-(2-hydroxyethoxymethyl)guanine on viral-specific polypeptide synthesis in human cytomegalovirus-infected cells. AB - Plaque-forming assay resulted in a 50 percent inhibitory dose by 9-(2 hydroxyethoxymethyl)guanine (acyclovir) against Towne strain human cytomegalovirus (HCMV) of approximately 98 mumol. At the drug concentration of 200 mumol, we did not detect any significant inhibition of viral DNA synthesis by cRNA-DNA hybridization. However, at this drug concentration, the synthesis of at least two viral-specific late polypeptides (150K and 67K) was significantly retarded up to 48 hours after infection, but resumed at 72 hours. These data suggest that acyclovir or its in vivo transformed derivative had a transient effect on viral-specific polypeptide synthesis in HCMV-infected human fibroblasts at a high drug concentration. PMID- 6285740 TI - Lymphocyte responses to herpes simplex virus isolates from patients with primary and recurrent herpes simplex genitalis. AB - The ability of herpes simplex (HSV) isolates from patients with herpes simplex genitalis (HSG) to induce lymphocyte transformation in cells from unrelated, healthy, seropositive donors was examined in a standard lymphocyte transformation assay. All HSV isolates were obtained, before the initiation of therapy, from patients who were enrolled in a placebo-controlled trial of acyclovir in the treatment of HSG. Isolates from patients with primary infections were used, as well as those from patients with frequent (eight or more episodes per year) or infrequent (two or fewer per year) recurrent disease. Blastogenic responses to isolates from patients with infrequent HSG recurrences were significantly less than those to isolates from patients either primary infections or frequent recurrences. No differences between the latter two groups of isolates were seen. These observations demonstrate that differences among naturally occurring HSV isolates exist as determined by this in vitro assay of host-virus interactions. Differences among strains may be important in the pathogenesis of HSV infections, particularly with respect to latency. PMID- 6285741 TI - The use of restriction endonucleases in the prenatal diagnosis of hemoglobinopathies. AB - Enzymes which fragment DNA at specific sites have been used to prenatally detect sickle cell anemia, alpha-thalassemia, and beta-thalassemia from amniotic fluid aspirates. Within the last few years, the use of combined restriction endonuclease systems (EcoRI and HindIII) for the antenatal detection of sickle cell anemia was found to provide an exact diagnosis of sickle cell status in 80% of couples at risk. Similar systems have been developed for homozygous alpha- and beta-thalassemia. Clinical applications of this technique and future implications are discussed. PMID- 6285742 TI - Early invasive and in situ warty carcinoma of the vulva: clinical, histologic, and electron microscopic study with particular reference to viral association. AB - Cases of carcinoma of the vulva with a warty appearance were reviewed. Altogether, 27 cases of warty carcinoma of the vulva were treated at the University of Minnesota Hospitals between 1976 and 1980, which accounted for 22.4% of all vulvar epithelial malignancies. As compared to the previous experience between 1951 and 1970, this relative frequency increased fourfold. Clinical history indicated that warty carcinoma of the vulva was not necessarily preceded by long lead periods of in situ lesions. The lesions seemed to occur in all ages after adolescence, were multifocal in one third of the cases, were frequently locally recurring, and were relatively benign despite their often large size. Many of them were originally diagnosed as condyloma acuminatum. Virus like particles were seen in eight of the 12 cases (67%) examined with transmission electron microscopy, which included six cases of invasive lesions. These observations suggest that human papillomavirus (HPV) may be the important etiologic agent of this group of tumors. Warty carcinoma of the vulva must be clearly separated from conventional invasive squamous cell carcinoma of the vulva, as its clinical behavior and a possible etiology appear unique. PMID- 6285743 TI - Latency of herpes simplex virus in uterosacral ligaments. PMID- 6285745 TI - Myeloperoxidase staining by the reagents of an automated differential analyzer (Hemalog-D) PMID- 6285744 TI - Effects of percutaneous estradiol and conjugated estrogens on the level of plasma proteins and triglycerides in postmenopausal women. AB - Percutaneous administration of estradiol (E2) is a new substitutive treatment for postmenopausal women. In order to compare hepatic action of percutaneous E2 with that of conjugated estrogens, 18 postmenopausal women were allocated at random to receive one of these two types of natural estrogens for 21 days. Eight patients (group I) received conjugated estrogens orally, 1.25 mg daily. Ten patients (group II) were told to apply percutaneous E2 ointment, 5 gm (i.e., 3 mg of E2), each evening on the abdominal skin. E2, estrone (E1), follicle-stimulating hormone (FSH), and luteinizing hormone (LH), and several markers of estrogen action were evaluated before and after treatment. Both types of treatment were biologically effective, as indicated by the decrease in plasma gonadotropins and the increase in estrogen levels. However, conjugated estrogens produced a greater increase in E1 than in E2; hence, the E2/E1 ratio was 0.57 in group I, whereas it was approximately 1 in group II. Plasma renin substrate increased significantly (by 180%) in group I but not in group II. In the same way, conjugated estrogens produced a modest (12%) but significant decrease in antithrombin III, whereas there was no variation with percutaneous E2. Sex steroid-binding protein was the most sensitive parameter for the hepatic action of estrogen, and increased by 18.66% with percutaneous E2 and by 150% with conjugated estrogens. Plasma triglycerides tended to increase in group I and to decrease in group II, but not significantly. Therefore, percutaneous administration of of E2, in contrast to conjugated estrogens, can produce plasma levels of estrogens closer to those observed in the follicular phase and less alterations in protein synthesis. This lesser toxicity may be explained partially by the route of administration, since with percutaneous administration of E2, the steroid bypasses the liver. PMID- 6285746 TI - Lymphocyte cyclic AMP levels antepartum and postpartum as an indirect assessment of the immune system during pregnancy. AB - The literature dealing with the immunological state of the pregnant woman has been conflicting. The concentrations and activity of a number of hormones and proteins which modify lymphocytic activity have been measured both in vivo and in vitro during pregnancy. Most of the differences between reported studies can be reconciled to technical or experimental variations. In some instances, the purported suppressive effects of embryonic proteins such as HCG have actually been caused by the impurity of the preparation studied. We have attempted to approach the question of whether or not the pregnant woman is immunosuppressed by studying a regulatory material in the lymphocytes. It is known that cAMP is a mirror of lymphocytic activity and that low levels of cAMP may indicate a high degree of reactivity, while high levels are present when lymphocyte reactivity is low. In an initial study, ten women volunteered to have blood drawn in the last trimester and two months postpartum. Cyclic AMP was extracted from lymphocyte enriched leukocytes and stored until all samples were available from all patients so that the analysis could be made simultaneously. Five samples were obtained from healthy nonpregnant women in the same age range. Postpartum and nonpregnant women were found to have significantly elevated levels of cAMP as compared to the lymphocytes obtained in the third trimester of pregnancy. The experiments were then repeated using seven more patients. The same significant increase in postpartum lymphocyte cAMP concentrations were found. The precise reason(s) for this is not known, but may be due to increased suppressor cell activity. PMID- 6285747 TI - Primary mediastinal carcinoid tumors. AB - A study of 15 cases of carcinoid tumor of the thymus, diagnosed and treated at the Mayo Clinic, revealed histopathologic features of the neoplasm that might cause it to be confused with other mediastinal tumors, both primary and secondary. The tumor was associated with ectopic ACTH production in six patients, of whom five had Cushing's syndrome. Metastasis, which occurred in 11 patients (73%), was delayed for as long as 8 years after initial diagnosis. Eleven patients underwent surgical resection of their primary thymic neoplasms, and nine of these received postoperative radiotherapy or chemotherapy or both. One patient was treated with chemotherapy alone, and three had no treatment for their neoplasms, which were discovered at autopsy. Eight patients are still alive. Of these, five have developed metastases, and only one is free of metastatic disease more than 5 years after diagnosis. In the two remaining cases, the discovery of the thymic tumor was recent and follow-up is not yet meaningful. Overall, four of the seven patients who died had proven metastatic disease. The mean survival after the appearance of extrathymic tumor in this group was 3 years. Of the five patients who are still alive with metastasis, all have survived at least 2 years since that spread of their disease. PMID- 6285748 TI - Primary nonlymphoreticular malignant neoplasms of the spleen. AB - Primary sarcomas of the spleen are exceedingly uncommon neoplasms, approximately 90 substantiated cases having been reported. We report on six cases of primary splenic angiosarcoma and three cases of primary malignant fibrous histiocytoma of the spleen which have been seen at the Mayo Clinic during the last 51 years. In one of the cases of splenic angiosarcoma, the lesion was manifested by spontaneous rupture and hemoperitoneum; interestingly, two cases of splenic malignant fibrous histiocytoma were of the inflammatory type and showed corresponding clinical features of a systemic nature. Five patients with angiosarcoma of the spleen died of metastatic tumor within 3 years after diagnosis; one patient with splenic malignant fibrous histiocytoma is alive with metastasis 11/2 years after splenectomy. One case of splenic angiosarcoma and two of malignant fibrous histiocytoma of the spleen have been diagnosed only recently. All three patients are alive and currently tumor-free, from 3 to 7 months after diagnosis. PMID- 6285750 TI - Vector competence of Culex pipiens quinquefasciatus for Murray Valley encephalitis, Kunjin, and Ross River viruses from Australia. AB - Australian populations of Culex pipiens quinquefasciatus Say from Brisbane, Charleville, Cairns, Kowanyama (Queensland), Darwin (Northern Territory), Mildura (Victoria), and Port Hedland (Western Australia) proved to be either poorly susceptible or refractory to oral infection with low passage level Murray Valley encephalitis, Kunjin, and Ross River viruses. With respect to past or future epidemics of Murray Valley encephalitis or Kunjin (both known to cause clinical encephalitis in man), and of epidemic polyarthritis, this mosquito is viewed as an unlikely vector. PMID- 6285749 TI - Dengue virus-specific and flavivirus group determinants identified with monoclonal antibodies by indirect immunofluorescence. AB - Monoclonal antibodies produced against the four dengue virus serotypes identified four categories of reactions by immunofluorescence: flavivirus group reactive, dengue complex specific, dengue subcomplex specific (DEN-1, DEN-3), and dengue serotype specific. This is the first time that monospecific antibodies have been available for all of these unique antigenic determinants. Hybridoma cell lines producing dengue type-specific antibodies have been deposited in the Hybridoma Cell Bank of the American Type Culture Collection (Rockville, MD) for general distribution. PMID- 6285751 TI - [Blood antidiuretic hormone, ACTH and prolactin during the administration of fentanyl in man]. PMID- 6285752 TI - [Studies on prolyl hydroxylase of bovine dental pulp (author's transl)]. PMID- 6285753 TI - [Neutrophil enzyme activity and their glycogen content in inflammatory diseases of the internal female genitalia]. PMID- 6285754 TI - Mechanism of ethanol-induced adrenal stimulation. AB - A historical review of studies addressing the adrenal cortical response to ethanol and the mechanisms thought to contribute to such responses is presented. Studies reviewed range from the earliest based upon crude bioassay techniques to more modern papers which have utilized highly specific radioimmunoassay techniques and isolated perfused rat adrenal glands. Evidence for both hypothalamic-pituitary as well as direct adrenal stimulation is presented. Available evidence, however, would suggest that acetaldehyde rather than ethanol per se, is responsible for direct adrenal stimulation when it occurs. PMID- 6285755 TI - The effect of alcohol ingestion on the susceptibility of mice to viral infections. AB - The influence of acute alcoholization on the evolution of different viral diseases was studied in orally alcoholized mice. Ethanol increased mice susceptibility to encephalomyocarditis and influenza only when it was administered after virus infection. There was no dose-effect correlation. Vaccinia and Herpes virus infections were not modified by alcoholization. Hypotheses concerning the mechanisms of alcohol action were put forward. PMID- 6285756 TI - 4-aminopyridine-a review. AB - 4-aminopyridine is the first of the aminopyridines to be used in clinical practice. It blocks potassium channels and thereby increases acetylcholine, and possibly noradrenaline, release at nerve terminals. In man the drug has a significant action at the neuromuscular junction, but has little effect on the autonomic nervous system or muscle (smooth, skeletal, or cardiac) although such actions have been demonstrated in animals. It may be clinically useful in the reversal of nondepolariser blockade and a role in antibiotic associated block has been proposed. It may be used effectively as an analeptic agent. It appears to be a useful therapeutic agent in myasthenia gravis and Eaton Lambert syndrome, although of limited use in botulism. It effects on the central nervous system are considerable. These account for the major side effects of the drug which include tremor, excitability and convulsions. PMID- 6285757 TI - Bronchial adenoma mimicking complication of endotracheal intubation. AB - A 39-year-old Chinese lady was admitted to hospital for an emergency caesarian section under general anaesthesia. After intubation it was discovered that there was diminished air entry to the left side. The cause could not be elicited during anaesthesia and a postoperative chest radiograph revealed no abnormality. Subsequent tomography revealed a mass in the lumen of the left main bronchus. Biopsy of the lesion revealed an adenoid cystic carcinoma (bronchial adenoma cylindroma type). By causing airway obstruction with gas trapping this rare neoplasm may present as the cause of a problem mimicking more common complications of endotracheal intubation. PMID- 6285758 TI - Assay of prolyl hydroxylase in cultured fibroblast monolayers. PMID- 6285759 TI - Synthesis and characterization of N-(4-azidophenylthio)phthalimide: A cleavable, photoactivable crosslinking reagent that reacts with sulfhydryl groups. PMID- 6285761 TI - Use of a novel H tube in transport studies with ionophores. PMID- 6285762 TI - Use of Percoll density gradient centrifugation for preparing isolated rat hepatocytes having long-term viability. PMID- 6285760 TI - Use of a new retrieving adaptor in the cloning of a synthetic human insulin A chain gene. PMID- 6285763 TI - Incorporation of lipids into cellular membranes--a spin-label assay. PMID- 6285764 TI - Rapid identification of Escherichia coli transformed by pBR322 carrying inserts at the PstI Site. PMID- 6285765 TI - Enzymatic conversion of deoxycytidine 5'-monophosphate to 5-methyldeoxycytidine 5'-triphosphate. PMID- 6285766 TI - Determination of polybrominated biphenyls in serum by negative chemical ionization mass spectrometry. PMID- 6285767 TI - Neuromuscular effects of atracurium in man. AB - The neuromuscular effects of atracurium were studied in 25 A.S.A. class I or II patients anesthetized by a N2O-O2 narcotic technique. In five patients incremental doses of 0.05 to 0.1 mg/kg of atracurium were given intravenously every 3 minutes until approximately 95% depression of the evoked electromyographic (EMG) response of the adductor policus muscle was produced. This required 0.25 to 0.35 mg/kg of atracurium. The duration of block (return to 95% of control) was 25 to 50 minutes. In addition, four groups of five patients each received 0.15, 0.25, 0.375, or 0.6mg/kg of atracurium. The block produced by 0.15 mg/kg was 10% to 92% and lasted 8 to 55 minutes. The block produced by 0.25, 0.375, and 0.6 mg/kg was 95% or greater with a duration of action of 30 to 68 minutes, 52 to 70 minutes, and 65 to 95 minutes, respectively. Tracheal intubation was easily carried out in all patients in whom there was a block of 90% or greater. The block could be antagonized by the common clinical combination of atropine and neostigmine. Changes in heart rate and blood pressure following atracurium were less than 5%. PMID- 6285768 TI - [Fine needle aspiration biopsy of lung cancers: a cytological and histological correlation]. PMID- 6285769 TI - [ACTH and cortisol content of the plasma and urinary excretion of corticosteroids before and after treatment of hyperthyroidism]. PMID- 6285770 TI - Frequency of bluetongue and bovine parvovirus infection in cattle in South Carolina dairy herds. AB - A survey of South Carolina diary cattle in 12 commercial herds was conducted, using serologic testing to determine the frequency and impact on reproduction of bluetongue (BT) and bovine parvovirus (BFV) infections. Results of the study of serum antibodies titers and dairy cattle health records indicated that the majority of the cattle surveyed was adequately protected against the major reproductive tract diseases through vaccination programs. The frequency of reproductive dysfunction was common in vaccinated herds, however. Six of the 12 herds in the survey had cattle which were BT serotest reactors, although total numbers of these reactor cattle were relatively small. Further, reproductive performance of reactor cows indicated that BT was of little consequence and was not associated with reproductive problems. All herds contained BPV serotest reactors and the latter comprised 59.7% of the total number of cows surveyed. The BPV seroreactor cows were commonly associated with the group of reproductive problem cows which experienced higher rates of embryonic mortality and more services per conception than did nonreactor cattle. The results of the survey emphasize the need for continued investigation and efforts to control BT, and especially BPV infection, in dairy cattle. PMID- 6285771 TI - Virologic study of the reproductive tract in dairy cows. AB - From Aug 1, 1979 to Aug 15, 1980, a total of 624 samples were collected from the vaginas, cervices, and uteri of 208 dairy cows 30 to 37 days after parturition. An additional 90 samples were collected from the reproductive tracts from 30 of the 208 cows. The 30 cows had aborted or were repeat breeders. Samples from cows that had aborted were collected within a week after clinical abortion or immediately after rectal palpation revealed subclinical abortion. All samples were tested for the presence of cytopathogenic viruses. All samples were negative. PMID- 6285772 TI - Demonstration of rotavirus antigen in trypsin-digested paraffin tissue sections by immunofluorescence. AB - Trypsin-digested paraffin tissue sections were used to demonstrate rotavirus particles in small intestinal epithelial cells of dirrheic neonatal calves, using direct fluorescent-antibody assay. The results were compared with other techniques to demonstrate rotavirus particles in tissue sections by immunofluorescence. Enzyme treatment of deparaffinized tissue sections gave excellent results with distinct specific fluorescence and minimal background. Prior staining of the tissue section with Mayer's hematoxylin made simultaneous observation of the same tissue section by light and ultraviolet microscopies possible. PMID- 6285774 TI - Immunocytochemical labeling of Anaplasma marginale Theiler in Dermacentor andersoni Stiles with peroxidase- antiperoxidase technique. AB - Colonies of Anaplasma marginale in midgut epithelial cells of adult ticks that had been infected as nymphs were specifically labeled, using the unlabeled antibody peroxidase-antiperoxidase method of immunocytochemistry. Visual comparison of infected and control tissue sections with the electron microscope demonstrated deposition of ring-like peroxidase-antiperoxidase complexes over organisms within the colonies. The intensity of labeling differed among organisms within a single colony, possibly as a result of varying antigenicity. The labeling observed on organisms in the colonies was similar to that seen on anaplasmal initial bodies in inclusions of infected bovine erythrocytes examined concurrently. PMID- 6285773 TI - Comparison of the enzyme-linked immunosorbent assay with an early polykaryocytosis inhibition assay and the agar-gel immunodiffusion test for the detection of antibodies to bovine leukemia virus. PMID- 6285775 TI - Decay of colostral antibodies to bovine leukemia virus with application to detection of calfhood infection. AB - An estimated weighted-regression method was used to describe the decay of colostral bovine leukemia virus (BLV) antibodies in the calf, as measured by agar gel immunodiffusion with glycoprotein antigen. The prediction equation, based on 473 observations from 130 animals, was log10 inverse titer = 1.29 -0.012 age (days). The half-life of BLV antibodies was estimated to be 25.8 days. Ages at which colostral antibodies were last detected were between 51 and 187 days. Normal limits of antibody decay were estimated and used to identify virus-induced active antibodies in calves during the colostral antibody period. Calves known to be infected were identified between 2 and 180 days of age, using 95% limits. Application of this procedure for the early serologic detection of BLV-infected calves in eradication or control programs is discussed. PMID- 6285776 TI - Validation of a rapid solid-phase radioimmunoassay for canine, bovine, and equine insulin. AB - A rapid and convenient commercial radioimmunoassay kit, developed for quantifying hormones in specimens from human beings, was validated for use in measuring insulin in serum of dogs, cattle, and horses. The procedure uses polypropylene assay tubes treated with rabbit anti-porcine insulin serum and porcine [125I]iodoinsulin. Specificity was proven by demonstrating that standard solutions of porcine insulin and serial dilutions of canine, bovine, and equine sera and pancreatic extracts inhibited binding of [125I]iodoinsulin to the antibody in a parallel manner. Gel-filtration chromatography of pancreatic extracts yielded a major peak of immunoreactive material that eluted identically with [125I]iodoinsulin. Immunoreactivity was not associated with fractions that contain larger and smaller molecular weight peptides (eg, proinsulin and C peptide, respectively). Biological specificity of the assay was shown by demonstrating increased insulin in serum after injection of glucose into heifers and glucagon into dogs and horses. Purified insulin and insulin in pancreatic extracts could be quantitatively recovered from serum, thereby demonstrating accuracy of the assay. Interassay precision of 5 control specimens run in 20 consecutive assays ranged from 6.7% to 20.1% (coefficient of variation) and intra assay precision of 6 control specimens each assayed 10 times ranged from 4.4% to 10.7% (coefficient of variation). Sensitivity of the assay was 3.2 microIU/ml. This radioimmunoassay for insulin is ideal for veterinary research and diagnosis, because a single set of reagents and procedures can be used for at least 3 species. PMID- 6285777 TI - Host defense systems in cattle exposed to polybrominated biphenyl. AB - In vivo and in vitro immunologic studies were undertaken in 114 Holstein cattle, 60 of which had detectable polybrominated biphenyls (PBB) in adipose tissue. The tissue PBB concentrations were between 0.02 and at least 1,000 mg/kg. An immunologic test profile was used to evaluate the influence of PBB on bovine immune competence. The profile included serum immunoglobulin determinations, autoantibody studies, lymphocyte subpopulation determinations, lymphoblastogenesis assays, delayed hypersensitivity skin testing, and neutrophil function studies. The results of the evaluation indicated that the host defenses are intact, both in quantity and function, in cattle with PBB body burdens ranging from 0.02 and 24.0 mg/kg for at least 2 years. Cattle fed 25 g of PBB daily during the acute toxicity study exhibited usual immune competence until they accumulated 500 g of PBB (tissue concentrations exceeding 1,000 mg/kg). Cattle that were fed more than 500 g of PBB became moribund and exhibited changes in neutrophil function tests and serum antibody titers. PMID- 6285779 TI - Effect of levamisole in immune responses to bovine herpesvirus-1. AB - The effect of levamisole on bovine immune responses to infectious bovine rhinotracheitis virus was assessed under laboratory and commercial feedlot situations. In all instances, levamisole appeared to have a beneficial effect on antibody responses of the cattle after vaccination. In the smaller scale pilot trials, levamisole appeared to be more efficacious when given 7 days after vaccination, presumably when a large amount of viral antigens was present as a result of viral replication. In the larger feedlot trial, however, response to administration of levamisole at the time of vaccination appeared to be slightly better than if given 7 days later. In all instances that animals had an antibody response before they were challenge-exposed with virulent virus, rectal temperature responses remained below 40 C, indicating that a threshold level of immunity may be acquired after the vaccination and that elevation of this threshold level does not necessarily alter the clinical disease. However, the amount of virus replication and shedding after challenge exposure seemed to be correlated with the level of immunity. These results are discussed in relationship to the role of immunity levels to spread of virus within a feedlot. PMID- 6285778 TI - Lymphocytes from ponies experimentally infected with equine herpesvirus 1: subpopulation dynamics and their response to mitogens. AB - Six pony foals, free of detectable serum neutralization (SN) antibody against equine herpesvirus type 1 by the standard virus-neutralization (VN) test, were inoculated with equine herpesvirus type 1. The ponies showed typical clinical signs of respiratory tract disease and developed a transient leukopenia, involving lymphocytes as well as neutrophils. The leukopenia reached its lowest point on postinoculation days (PID) 3 to 5 and then returned to base-line values by PID 8 to 10. On quantitation of lymphocyte subpopulations, T and B lymphocytes were decreased during the onset of leukopenia and then recouped during the recovery from leukopenia. However, the proportions of the T and B lymphocytes remained constant during the lymphopenia, ranging from 70% to 80% and 20% to 30%, respectively. The lymphocyte blastogenic response to mitogens increased to peak by PID 2 to 5 and then decreased to base line values or below by PID 7. Mitogen responses of T lymphocyte and mixed lymphocyte preparations were nearly similar. However, the responses of 2 ponies wee somewhat different from the responses of others in that there was an increase in the B lymphocytes in the range of 40% to 50% during the recovery phase of lymphopenia. Also, the 2 ponies' mixed lymphocyte response to mitogens was considerably higher and the T lymphocyte response to mitogen was lower as compared with that of mixed lymphocyte preparation. The importance of these 2 types of responses is discussed. PMID- 6285780 TI - Association between route of inoculation with infectious bovine rhinotracheitis virus and site of recrudescence after dexamethasone treatment. AB - Four calves latently infected with infectious bovine rhinotracheitis virus (IBRV) were used to compare the ease of isolation of virus from neuronal ganglia and from mucosal surfaces. Two calves were slaughtered, and neuronal ganglia (cranial cervical, trigeminal, and 3rd and 4th sacral) were cocultivated on bovine fetal kidney cells. Virus was not isolated. Two calves given dexamethasone for 4 days were slaughtered on the 5th day. Virus was not isolated from cocultivated or macerated neuronal ganglia, but virus was isolated from nasal secretions taken from both calves on the day of slaughter. Eleven calves were inoculated with IBRV via different routes and were treated with dexamethasone 3 to 4 months after inoculation. virus was isolated from the nasal cavities, but not the vaginas of 6 heifers inoculated intranasally, and was isolated from the vaginas, but not the nasal cavities of 2 heifers inoculated intravaginally. Of 3 calves inoculated IV, virus was isolated from the nasal cavities of 3, from the oropharynxes of 2, and from the prepuce of 1. PMID- 6285781 TI - Prevalence of bovine herpesvirus-1, bovine oral diarrhea, parainfluenza-3, bovine adenoviruses-3 and -7, and goat respiratory syncytial viral antibodies in goats. AB - Sera from healthy goats were collected during October 1979 through October 1980. These sera were tested for bovine herpesvirus-1 (BHV-1), bovine viral diarrhea virus (BVDV), parainfluenza-3 (PI-3) virus, bovine adenoviruses (BAV) -3 and -7, and goat respiratory syncytial virus (GRSV) antibodies by microtitration virus neutralization test. The number of herds with seropositive goats for each virus were: 5/38 (13.2%) for BHV-1; 9/38 (23.7%) for BVDV; 8/38 (21.1%) for PI-3 virus; 1/38 (2.6%) for BAV-3; 15/38 (39.5%) for BAV-7; and 26/34 (76.5%) for GRSV. Seropositive rates for each virus for the individual goats tested were: 6/502 (1.2%) for BHV-1; 9/498 (1.8%) for BVDV; 49/458 (10.75) for PI-3 virus; 1/487 (0.025) for BAV-3; 40/448 (8.9%) for BAV-7; and 166/332 (50.0%) for GRSV. PMID- 6285782 TI - Role of insects in the transmission of bovine leukosis virus: potential for transmission by mosquitoes. AB - Bovine leukosis virus (BLV) was transmitted to sheep in a simulated mechanical transmission experiment, using the following species of mosquitoes; Anopheles freeborni, A stephensi, A quadrimaculatus, and A albimanus. Mosquitoes were fed on blood taken from a BLV-infected cow with persistent lymphocytosis. Mouthparts and heads of mosquitoes were removed immediately after feeding, placed in RPMI 1640 medium, and inoculated subcutaneously into sheep. Nine sheep were inoculated with mouthparts and heads from 37 to 122 mosquitoes. Infection was determined serologically. Three monthly serum samples were collected from the sheep and were tested for the presence of antibodies to BLV, using the agar-gel immunodiffusion (AGID) test. Sera that were negative by AGID at 3 months were tested by radioimmunoassay. Results from radioimmunoassay agreed with those obtained by AGID. Four of the 9 sheep developed antibody to BLV. Sheep that seroconverted were inoculated with mouthparts and heads from as few as 54 mosquitoes. PMID- 6285784 TI - Border disease: experimental reproduction in sheep, using a virus replicated in tissue culture. PMID- 6285783 TI - Border disease: tissue culture studies of the virus in sheep. AB - A California strain of border disease (BD) virus was isolated from the CNS and spleen of an infected lamb and was propagated without cytopathic effect on 2 tissue culture systems. Viral antigen was detected in the cytoplasm of infected cells by immunofluorescence. The growth curve of this BD virus in primary sheep choroid plexus cells revealed a latent period of 12 hours and peak viral production by 36 hours after inoculation. The growth curve in porcine kidney cells (PK-15) demonstrated a latent period of 8 hours and peak viral production by 54 hours after inoculation. The primary sheep choroid plexus cell tissue culture isolate of BD virus was then used to develop hyperimmune sera in 2 adult sheep. Neutralizing antibody was not detected until 43 days after primary inoculation when a slow antibody increase occurred in both sheep. PMID- 6285785 TI - The release of elastase, myeloperoxidase, and lysozyme from human alveolar macrophages. AB - Human alveolar macrophages from lungs of cigarette smokers were retrieved by lavage of surgical specimens. The macrophage secretions were harvested after 18 h of incubation. The medium contained at least 2 acid-stable factors that could release enzymes from cytochalasin-B-treated human neutrophils. Our study focused on the largest of these factors, which had an apparent mass ratio of 5,400 by gel filtration chromatography in 10% acetic acid. The high molecular weight (HMW) factor was partially degraded by trypsin. Chymotrypsin completely destroyed the factor, but human neutrophil elastase did not affect it. The factor is partially extractable into chloroform indicating that it is very hydrophobic and may contain a lipid. High concentrations of the HMW factor inhibited the release of lysozyme and myeloperoxidase. Because elastases can cause emphysema when introduced into alveoli of animals, the most important observation may be that the HMW factor was able to release elastase from human neutrophils attached to Millipore membranes in the absence of cytochalasin B. The enzyme-releasing factors may be identical to neutrophil chemotactic factors recently described by others. The contribution of the released elastase to the protease load in the lung may be augmented by the simultaneous release from neutrophils of myeloperoxidase, which can inactivate alpha 1-antitrypsin. This interaction between alveolar macrophages and neutrophils may have importance in the pathogenesis of emphysema. PMID- 6285787 TI - [Acute gastroenteritis caused by rotavirus (author's transl)]. AB - A prospective study of 87 children hospital admitted for acute gastroenteritis [AGE] with ages between 24 days and three years and a control group of 32 children with ages between six months and three years, is presented. In all cases a virological study of stools is made by electron microscopy [E/M]. In 63 percent of the 87 patients with AGE, viruses were isolated, being rotavirus the most common, 45.9 percent, a similar incidence as that found in developed countries. The clinical course and analytical findings within the groups with viral and non viral AGE do not show significant differences. Special stress is set upon the reliability of the E/M technique on the stools for rotavirus induced AGE diagnosis, due to the good correlation with clinical data, presence of seroconversion and the finding of virus in duodenal juice. Results of intestinal biopsy in a certain group of children are shown. PMID- 6285786 TI - Demonstration of a formyl peptide receptor on lung macrophages: correlation of binding properties with chemotaxis and release of superoxide anion. PMID- 6285788 TI - Late-onset 21-hydroxylase deficiency. PMID- 6285789 TI - Ketoconazole effects. PMID- 6285790 TI - [ACTH, beta-endorphin, lipotropins, and other pro-opiocortin-derived peptides: a new family of hormones (author's transl)]. AB - beta-Endorphin, a pituitary morphino-mimetic peptide, was identified in a culture medium derived from a human corticotropic adenoma. Secretion products from cultured human cells derived from a small-cell carcinoma of the lung were shown to contain a high molecular weight precursor analagous to pro-opiocortin: this molecule is a polypeptide of the order of 28,000 daltons the enzymatic processing of which leads to the coordinated and simultaneous release of different peptide fragments: ACTH, beta- and gamma-lipotropins, beta-endorphin, fragment 16 K and gamma 3-MSH. All these peptides have been identified in human plasma, and pituitary and non-pituitary tumor extracts. Their plasma concentrations vary in a parallel manner, beta-endorphin and a peptide very similar to beta-MSH have been detected in human hypothalamus. PMID- 6285791 TI - [ACTH, beta-endorphin and lipotropins: physiopathological studies in man (author's transl)]. AB - ACTH and lipotropins (beta- and gamma-LPH) are synthesized from a common precursor by the pituitary corticotropic cell. We have measured LPH plasma levels under physiological and pathological conditions and we have compared them with ACTH plasma levels in the same circumstances. Spontaneous variations (nycthemeral rhythm) in LPH, ACTH and cortisol plasma levels were parallel, while responses to Dexamethasone freination test and stress (Insulin induced hypoglycemia) or more specific stimulation (Metopirone, lysine-vasopressin) were parallel and superimposable. LPH levels were always higher than ACTH levels in two pathological circumstances: chronic renal failure and Cushing's syndromes with ectopic ACTH producing tumors. The determination of both ACTH and LPH levels assists the diagnosis of corticotropic insufficiency and etiologic investigation of Cushing's syndrome, after hypercorticolism had been established. Although unable to confirm the presence of corticotropic adenoma in patients with Cushing's disease, or the predict effectiveness of pituitary surgery, these determination bring good arguments for treated Cushing's diseases follow up. PMID- 6285792 TI - [Isolated ACTH deficiency associated with leukoneutropenia (author's transl)]. AB - The paper describes the case of a 38 year's old alcoholic patient. The investigation of his severe asthenia led to the discovery of an isolated ACTH deficiency: plasma cortisol was less than 1 microgram 100 ml reactivable by tetracosa-peptide beta 1-24. Plasma ACTH was low and remained so after LVD and metyrapone. Persistent leuconeutropenia without myelogram and agar cultures abnormalities was noted. Etiocholanolone test was negative and epinephrine induced a significant rise in blood leucocytes. The authors review the mechanisms of corticoid action on leucopoiesis. After 10 months corticoid treatment, leuconeutropenia improved but was still present suggesting an other underlying mechanism than cortisol insufficiency per se. PMID- 6285793 TI - [Antagonistic effect against "Clostridium perfringens" exerted in the digestive tract of gnotobiotic mice by "Clostridium" strains isolated from the microflora of conventional mice (author's transl)]. AB - Bacterial strains isolated from the digestive tract of conventional mice produced a barrier effect against a strain of Clostridium perfringens type A (strain CP) in the gastrointestinal tract of gnotobiotic mice. This barrier effect was observed when mice, monoassociated with Escherichia coli K12, were inoculated with a mixed culture of fusiform-shaped clostridia. The collection of fusiforms (collection D) was obtained from a single colony chosen from among the 26 types studied. Collection D and E. coli K12 together exerted a barrier effect against all strains of C. perfringens type A tested. No effect was observed against a C. perfringens type C strain. The expression of the barrier effect depended on the order in which the strains were used to inoculate the mice. Thus strain CP was eliminated from the digestive tract when the mice had previously been associated with collection D and E. coli K12. If the mice were inoculated with strain CP first, however, the barrier effect was only partial. We have also been able to use two strains of clostridia (C1, C3) instead of E. coli K12 to produce a drastic barrier effect against strain CP. The order in which the strains were used to inoculate the mice did not, in this case, influence the production of a barrier effect. Three strains (Da, Db, Dc) isolated from collection D, produced only a partial barrier effect against strain CP in mice previously associated with E. coli K12. These results illustrate the difficulties encountered in determining the minimal number of bacterial strains involved in the production of a barrier effect. PMID- 6285794 TI - [Two further cases of rare laryngeal tumors. Lipoma and chemodectoma (author's transl)]. AB - The cases of laryngeal tumors requiring surgical excision are presented. The first case was that of patient with a laryngeal lipoma of atypical appearance, histological findings suggesting a hamartoma. The second patient had a chemodectoma, diagnosis being difficult but facilitated by super-selective arteriography, requiring delicate excision which was almost bloodless as a result of previous embolization. Clinical and pathological features of these tumors reviewed, and problems related to their treatment discussed. The authors have previously reported 4 other rare types of laryngeal tumor. PMID- 6285795 TI - [Meningioma of the ear simulating a glomus tumor: report on two cases (author's transl)]. AB - Two further cases of a meningioma of the ear simulating a glomus tumor are reported, pathological features in the first case leading to intensive discussion. Two such cases had been previously described in great detail in a paper published in 1977. In those patients, the differential diagnosis froma jugular glomus tumor was only established two and a half years later in one case, when excision was necessary because of an intracranial recurrence, and following re-examination of all sections of jugular glomus tumors previously operated upon, in the other case. Factors enabling detection of meningiomas simulating glomus tumors were also discussed in the original report. Theses included discordance of clinical finding, -minimal signs on arteriography, -the primordial value of contrast computed tomography, -systematic alerting of the pathologist. PMID- 6285796 TI - [Antiviral activity of culture filtrates of some Agaricales on the experimental infection with Coxsackievirus B (author's transl)]. PMID- 6285797 TI - [Dopamine and pituitary hormones. Physiology and pathology (author's transl)]. AB - Dopamine regulates the secretion of pituitary hormones: it inhibits permanently the production of prolactin and blocks the gonadotrophins and the thyroid stimulating hormone. It stimulates inconstantly the secretion of growth hormone and it does not control corticotropin. Dopamine agonists and antagonists are currently used in the investigation and the treatment of hyper-prolactinemia and acromegaly. PMID- 6285799 TI - Histological types of nasopharyngeal carcinoma as compared to EBV serology. AB - One hundred and thirteen cases of nasopharyngeal carcinoma (NPC) were classified histologically according to the WHO-, French and Cologne systems. The various histological tumor types were then correlated with data on Epstein-Barr virus (EBV) serology (EA, VCA, EBNA, CF, IgA-VCA). The results showed non-keratinizing carcinomas with lymphoid infiltration, and undifferentiated carcinomas to be associated with significantly elevated anti-EBV titers. These two tumor types can be easily grouped together according to the French classification scheme as "undifferentiated carcinoma of the nasopharyngeal type (UCNT)". Such a procedure, which may have therapeutic implication, simplifies the rapid diagnostic screening of NPC patients and also may enhance the reproducibility of histological tumor typing. PMID- 6285798 TI - [Clinical and laboratory features of endemic goiter in the Man region (Ivory Coast) (author's transl)]. AB - An epidemiological study involving 616 subjects carried out in the Man region, Cote d'Ivoire, showed endemic goiter in 54,5% of the group overall and in 80% of females. Thyroid function of subjects chosen at random in the goitrous and non goitrous group (105 G and 71 NG respectively) was identical. Compared with a French control group T4 was significantly lower but T3 and TSH were higher. TBG level is high with low iodine elimination. High levels of TSH are not necessarily associated with low T3 or T4 levels, or with clinical signs of a hypothyroid condition. They are probably the result of a transient reaction. T4-TSH and T3 TSH correlations for NG subjects were (-0,40) but the T3-TSH correlation for the G subjects was significantly lower (-0,23). These differences could be explained by the simultaneous high levels of T3 and TSH more frequently rencountered in G subjects and by the fact that serious hypothyroid conditions are more frequent in the NG group (6 out of 9). The percentage of hypothyroid conditions is high in both clinical and biological investigations. No anti-thyroglobulin antibodies were found in any of the hypothyroid cases with goitre. The histograms illustrate the clinical and biological heterogeneity of a homogeneous non-medicalised population. PMID- 6285800 TI - Acute conduction block associated with experimental antiserum-mediated demyelination of peripheral nerve. AB - Intraneural injection of antisera from rabbits with high antigalactocerebroside antibody levels into rat sciatic nerve produced acute nerve conduction block. This was first apparent in some motor axons between 30 and 60 minutes after injection and progressed to completion within 2 to 4 hours. Concurrent morphological evidence of demyelination was present, but structural changes at the time of onset of block were mild and were restricted to the myelin and Schwann cell, particularly at the paranodal areas and Schmidt-Lanterman clefts. It is suggested that paranodal lesions could account for the observed conduction block. PMID- 6285802 TI - Myelin and Schwann cell abnormalities in Isaac's syndrome. PMID- 6285803 TI - Proton beam therapy. PMID- 6285805 TI - [Long-term decomposition of bones. 1. Mineral phase]. AB - Based on the alteration of microstructure and gross features caused by long term fate of bony substance a model of decomposition of bone mineral is outlined. Destruction of the corps by microorganisms leads on the whole to an acid milieu. Hence hydroxyapatite is turned into brushite, which is more soluble in acid media. As this mineral is formed in a space consuming way, the expanding brushite masses support mechanical destruction of bony substance by cracking the lamellary systems. Both mechanical stress by formation of brushite and transformation of hydroxyapatite to this mineral compound are main features for dead bone decomposition. PMID- 6285801 TI - Macroglobulinemia with peripheral neuropathy simulating motor neuron disease. AB - A 48-year-old man with an IgM plasma cell dyscrasia died after 14 months of symptoms and signs typical of motor neuron disease, including widespread fasciculation and normal sensation. Two laboratory results were atypical: cerebrospinal fluid protein content of 132 mg/dl and slow motor nerve conduction. At autopsy, no loss or atrophy of anterior horn neurons was found; instead, degeneration of ventral and dorsal roots and retrograde changes of chromatolysis in motor neurons implied peripheral neuropathy. Most reported cases of neuropathy associated with plasma cell dyscrasias have been sensorimotor or purely sensory, but there have been 14 previous cases of motor disorders. PMID- 6285804 TI - The role of microwave frequency in EPR spectroscopy of copper complexes. PMID- 6285806 TI - Endogenous opiates and pain. PMID- 6285807 TI - [Experimental study of the endolymphatic use of gentamycin and klaforan (cefotaxime)]. AB - Distribution of gentamycin and klaforan in the lymphatic system, blood and cerebrospinal fluid, as well as their effect on lymph node morphology was studied experimentally on 46 dogs. The antibiotics were administered endolymphatically in doses of 1 and 30 mg/kg respectively. The highest levels of gentamycin (200-250 micrograms/ml) and klaforan (up to 1600 micrograms/ml) were detected in the central lymph. The therapeutic levels of the antibiotics in the central persisted for 24 and 72 hours respectively. The antibiotics accumulated mainly in the regional (inguinal and pelvic) lymph nodes, where their high levels were determined for 72 hours. The levels of the antibiotics in the distant (cervical and tracheobronchial) lymph nodes were lower and did not exceed the therapeutic ones. They persisted for 6 and 24 hours respectively. No unfavourable effect of the antibiotics on the structure and cell composition of the lymph nodes was noted. After endolymphatic administration in the above doses the antibiotic levels in the blood serum were close to those observed after administration by the routine routes. After endolymphatic administration in the above doses the antibiotics penetrated into the cerebrospinal fluid. PMID- 6285809 TI - Efficacy of ceftriaxone in serious bacterial infections. AB - Ceftriaxone is a new semisynthetic cephalosporin with broad-spectrum in vitro activity and an unusually long serum half-life. The clinical efficacy of ceftriaxone was evaluated in 35 infections in 34 patients; 12 of these patients had skin and soft tissue infections, 10 had infections of the urinary tract, 8 had pneumonia, 2 had biliary tract infections, 1 had sinusitis, 1 had diverticulitis, and 1 had a retroperitoneal abscess. Of the 35 infections, 9 were bacteremic. The bacteria isolated included Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus pneumoniae, Streptococcus faecalis, other streptococcal species, Escherichia coli, Proteus mirabilis, Klebsiella pneumoniae, Serratia marcescens, Enterobacter cloacae, Haemophilus influenzae, Pseudomonas aeruginosa, Bacteroides fragilis, other Bacteroides species, and anaerobic cocci. Improvement or cure occurred in 32 episodes, for a response rate of 91%. There were three treatment failures in patients with soft tissue infections. No serious drug toxicities were observed. At a dosage regimen of 1 g every 12 h the peak and trough serum antibiotic concentrations were well above the minimal inhibitory concentrations of most pathogens. Our findings suggest that ceftriaxone is a safe and effective antibiotic for therapy of serious bacterial infections. PMID- 6285808 TI - Effect of systemic antimicrobial prophylaxis on microbial flora. AB - Fifteen patients undergoing intensive chemotherapy for oat cell carcinoma of the lung in a protected environmental unit received antimicrobial prophylaxis with oral trimethoprim-sulfamethoxazole and short courses of parenteral ticarcillin, tobramycin, and miconazole. Altogether, 58 (65%) of 89 strains of aerobic bacteria and 28 (60%) of 47 strains of anaerobic bacteria present before prophylaxis were no longer cultured from stool specimens during prophylaxis. Ten strains of bacteria and four fungi were acquired in the stools during prophylaxis. Most fungi persisted in the throat and stools during prophylaxis. Bacterial infections occurred infrequently, but three patients developed Candida esophagitis. The regimen was well tolerated with minimal side effects. PMID- 6285810 TI - Cefotaxime therapy of serious bacterial infection in adults. AB - We evaluated the efficacy, safety, and tolerance of cefotaxime in 35 adults (25 with pleuropulmonary infections, 7 with genitourinary tract infections, and 3 with soft tissue infections). Of these 35 patients, 18 (51.4%) were seriously or critically ill. In vitro susceptibility testing revealed that 90.4% of the pathogens isolated were susceptible to cefotaxime (minimal inhibitory concentration, less than 8 micrograms/ml), 4.8% were intermediately susceptible (minimal inhibitory concentration, 8 to 32 micrograms/ml), and 4.8% were resistant (minimal inhibitory concentration, greater than 32 micrograms/ml). A total of 34 of the 35 patients (97%) were clinically and bacteriologically cured of their infections. Adverse reactions occurred in two patients, but these reactions did not require interruption of therapy. PMID- 6285811 TI - Inhibition of herpes simplex virus replication by succinyl concanavalin A. AB - Incubation of herpes simplex virus type 1-infected cells with succinyl concanavalin A, a derivative of the jack bean lectin concanavalin A, resulted in the decreased production of virus. The mode of inhibition by the lectin was unclear. No effect was apparent on the level of viral DNA synthesis. However, incubation of infected cells with increasing concentrations of the lectin appeared to result in a decrease in the quantity of viral protein produced within the cell. A reduction in the virus titer of 57 to 64% was observed upon direct incubation of extracellular virus in the presence of 50 to 100 micrograms of succinyl-concanavalin A per ml. PMID- 6285812 TI - Treatment of infections caused by ampicillin-resistant pathogens with a combination of ampicillin and CP-45,899. AB - Eleven patients infected with ampicillin-resistant organisms were treated with a combination of CP-45,899, a beta-lactamase inhibitor, and ampicillin. All infections were controlled within 7 days without any signs of toxicity. PMID- 6285813 TI - Conjugative R plasmids in Streptococcus faecium (group D). AB - Ten isolates of Streptococcus faecium were found to be resistant to penicillin, tetracycline, macrolides and related drugs, streptomycin, and kanamycin, and four strains were resistant to chloramphenicol. Six of these 10 strains transferred all their resistance markers (except penicillin) by conjugation at a low frequency (10(-7) to 10(-9)). Several plasmids of different molecular weights were found in each of the wild-type strains. In 5 of 11 transconjugant strains, R plasmids were detected which had molecular weights identical to those of the plasmids found in the corresponding donor strain. Each of the six other transconjugants harbored one plasmid with a size different from those found in the corresponding donor strain, suggesting the occurrence of molecular events during or after conjugative transfer. None of the five tetracycline-resistant transconjugants contained detectable satellite DNA, HindIII restriction enzyme fingerprints of S. faecium resistance plasmids were different from the HindIII patterns of macrolide, aminoglycoside, and tetracycline resistance plasmids from other strains of streptococci. PMID- 6285814 TI - Impairing effect of food on ketoconazole absorption. AB - Single oral doses of ketoconazole (200 mg) or miconazole (250 mg) were given in a randomized cross-over study to 10 healthy volunteers. Ketoconazole with administered (i) after fasting (both brand 1 [Orion Pharmaceutical Co.] and brand 2 [Janssen Pharmaceutica] were tested), (ii) after a standardized meal (660 kilocalories; 2,772 kJ) (brand 1), and (iii) with 300 ml of orange juice (pH 3.8) (brand 1). Miconazole was administered after fasting. Venous blood samples for high-performance liquid chromatography determinations of ketoconazole and gas chromatographic analyses of miconazole were drawn periodically up to 24 h. The concentrations of ketoconazole in sera attained with the two brands were not statistically different. The peak concentrations of ketoconazole attained with brand 1 were 4.1 +/- 0.3 micrograms/ml (mean +/- standard error of the mean) after fasting, 2.3 +/- 0.3 micrograms/ml after the standardized meal (P less than 0.01), and 3.6 +/- 0.2 micrograms/ml with orange juice. The peak concentrations were reached in 1.4, 2.3 (P less than 0.05), and 1.8 h, respectively, whereas the areas under the serum concentration-time curves were 14.4 +/- 2.21, 8.6 +/- 1.33 (P less than 0.05), and 13.4 +/- 1.30 micrograms.h/ml, respectively. The half lives (1.7 to 2 h) did not vary significantly among the different regimens. Compared with ketoconazole, oral absorption of miconazole was poor (peak concentration, 0.47 +/- 0.7 micrograms/ml; time to reach the peak concentration, 2.6 h; area under the serum concentration-time curve, 1.10 +/- 0.20 micrograms.h/ml). PMID- 6285815 TI - Effects of moxalactam and cefotaxime on rabbit renal tissue. AB - To determine and compare the effects of moxalactam and cefotaxime on kidneys, we gave these drugs in doses of 750 and 1,500 mg/kg to rabbits for 7 days. Cephaloridine was included as a positive control. Neither moxalactam nor cefotaxime at either dose caused lysosomal enzymuria, changes visible by light microscopy or increased plasma creatine. Both drugs caused minor alterations in glomerular ultrastructure at the higher dose. Cephaloridine, on the other hand, caused widespread renal functional and morphological damage. We conclude that in rabbits, both moxalactam and cefotaxime are remarkably nonnephrotoxic. PMID- 6285816 TI - Capillary agar diffusion assay for measuring metronidazole in human gingival crevice fluid. AB - An agar diffusion assay in capillaries, with Fusobacterium nucleatum M2 as the indicator strain, has been developed to measure metronidazole concentrations in gingival crevice fluid. The assay allows the measurement of 1.9 micrograms of metronidazole per ml in sample amounts of 0.6 microliters. PMID- 6285817 TI - In vitro susceptibility of Clostridium difficile isolates to cefotaxime, moxalactam, and cefoperazone. AB - The in vitro susceptibility of 20 isolates of Clostridium difficile to cefotaxime, moxalactam, and cefoperazone was determined by a standard agar dilution method. The median minimal inhibitory concentrations were 64, 32, and 32 mug/ml for cefotaxime, moxalactam, and cefoperazone, respectively. PMID- 6285819 TI - Pathological findings of adenine arabinoside (ARA-A) and cytarabine (ARA-C) in the treatment of herpes simplex encephalitis in rabbit model. AB - The injection of herpes simplex virus type 1 (HSV-1) into the vitreous body of the eye in the 18 day old albino rabbits consistently induced herpes encephalitis with 90% survival. In the untreated rabbits the lesions follow a defined anatomical pathway producing a progressive disease not dissimilar to the natural human disease in that HSV travels slowly by cell-to-cell infection of neuroglia. The effects of adenine arabinoside (ara-A) and cytarabine (ara-C) on HSV encephalitis in rabbit model were studied by starting the treatment on 4th day post-inoculation of HSV. Deaths due to toxic side effects were caused by ara-A and ara-C in 30% and 50% of animals respectively, compared with 10% in untreated animals. Neurological signs, such as head jerking, ataxia and frequent epileptiform fits, occurred in ara-A, and ara-C and untreated rabbits. Comparative histological studies of optic nerves and brains showed that ara-A and ara-C had no beneficial effect, but surprisingly enhanced the disease. PMID- 6285818 TI - Efficiency and selectivity of (E)-5-(2-bromovinyl)-2'-deoxyuridine and some other 5-substituted 2'-deoxypyrimidine nucleosides as anti-herpes agents. AB - A number of novel 5-substituted 2'deoxypyrimidine nucleosides exhibited antiviral activity against herpes simplex virus type 1 strain V3 (HSV-1-V3) when assayed under one-step conditions in primary human lung fibroblast j(PHLF) cell cultures, and compared with the reference compounds cytosine arabinoside (ara-C), 5-iodo-2' deoxyuridine (IUdR), and 5-iodo-5'amino-2',5'-dideoxyuridine (AIU). The most effective of these were (in order of decreasing activity): (E)-5-(2-bromovinyl) UdR (BrVUdR) greater than ara-C greater than IUdR greater than 5-azidomethyl-UdR (AMeUdR) greater than 5-formyl-UdR (fUdR) greater than 5-hydroxymethyl-UdR (HMeUdR) greater than AIU greater than 5-mercaptomethyl-UdR (MMeUdR) = 5 hydroxymethyl-2'-deoxy-cytidine (HMeCdR) greater than 5-benzyloxymethyl-UdR (BOMeUdR). In a multistep virus replication experiment (plaque reduction assay on Vero cells) the order of decreasing activity was as follows: BrVUdR = ara-C greater than HMeUdR greater than fUdR IUdR greater than HMeCdR greater than BOMeUdR greater than AMeUdR greater than AIU greater than MMeUdR. BrVUdR effected a 50% reduction in plaque formation of different strains of HSV-1 at a concentration of 0.06-0.22 microM, of pseudorabies virus (PRV) at 0.02-0.23 microM, and of herpes simplex virus type 2 (HSV-2) at 8 microM, whereas the ID50 values for adenovirus type 2 and type 5 were 100 and 50-100 microM, respectively. The growth of synchronied baby hamster kidney cells in suspension cultures was inhibited by 50% at concentrations of 100, 70, 20, 4, 8, and 0.2 microM for BrVUdR, HMeCdR, IUdR, fUdR, BOMeUdR, and HMeUdR, respectively. PMID- 6285820 TI - Restriction endonuclease analysis of the lactose plasmid in Streptococcus lactis ML3 and two recombinant lactose plasmids. AB - We investigated the molecular relationship between the 60-megadalton (Mdal) recombinant lactose plasmids in ML 3 x LM2301 lactose-positive (Lac+) transconjugants and the genetic material of Streptococcus lactis ML3. Lactose metabolism is linked to the 33-Mdal plasmid pSK08 in ML3, and the recipient LM2301 is cured of plasmid DNA. The plasmids were analyzed with a series of restriction enzymes. We found that the 60-Mdal plasmids of Lac+ transconjugants contained pSK08 DNA, but were not simply dimers of pSK08. The 60-Mdal plasmids contained a segment of DNA not apparent in pSK08. The restriction patterns of the 60-Mdal plasmid in a Lac+ nonclumping transconjugant and that in a Lac+ clumping transconjugant were different. This suggested that there was a molecular differences between these two recombinant plasmids. We conclude that the segment of DNA in the 60-Mdal plasmids that was not present in pSK08 was the proposed transfer factor responsible for cell aggregation and high-frequency conjugation. PMID- 6285821 TI - Effect of free-radical scavengers on enumeration of thermally stressed cells of Staphylococcus aureus MF-31. AB - Exposure of crude cell lysates of Staphylococcus aureus MF-31 to 5 or 10 mM hydrogen peroxide resulted in a linear decrease in superoxide dismutase activity. Approximately 13% of the superoxide dismutase activity was lost after 16 min. Thermally stressed and nonstressed cells were exposed to a photochemically generated exogenous flux of superoxide radicals (O2.-). The death of thermally stressed cells was linear with time. Addition of superoxide dismutase or catalase to the O2.- generating system resulted in protection of thermally stressed and nonstressed cells, with the protective effect being greater for thermally stressed cells. Incorporation of O2-, hydroxyl radical, or singlet oxygen scavengers or antioxidants to tryptic soy agar containing 7.5% NaCl did not increase the enumeration of thermally stressed cells. PMID- 6285822 TI - Inactivation by gamma irradiation of animal viruses in simulated laboratory effluent. AB - Several animal viruses were treated with gamma radiation from a 60Co source under conditions which might be found in effluent from an animal disease laboratory. Swine vesicular disease virus, vesicular stomatitis virus, and blue-tongue virus were irradiated in tissues from experimentally infected animals. Pseudorabies virus, fowl plague virus, swine vesicular disease virus, and vesicular stomatitis virus were irradiated in liquid animal feces. All were tested in animals and in vitro. The D10 values, that is, the doses required to reduce infectivity by 1 log10, were not apparently different from those expected from predictions based on other data and theoretical considerations. The existence of the viruses in pieces of tissue or in liquid feces made no difference in the efficacy of the gamma radiation for inactivating them. Under the "worst case" conditions (most protective for virus) simulated in this study, no infectious agents would survive 4.0 Mrads. PMID- 6285823 TI - Evidence that microorganisms cause inactivation of viruses in activated sludge. AB - Virus loss in activated sludge appeared to be caused by microorganisms. This conclusion is supported by the finding that poliovirus infectivity decreased during incubation in mixed-liquor suspended solids, primarily because of a sedimentable, heat-sensitive component. Furthermore, broth spiked with mixed liquor suspended solids acquired antiviral activity during incubation. PMID- 6285824 TI - Wet and dry bacterial spore densities determined by buoyant sedimentation. AB - The wet densities of various types of dormant bacterial spores and reference particles were determined by centrifugal buoyant sedimentation in density gradient solutions of three commercial media of high chemical density. With Metrizamide or Renografin, the wet density values for the spores and permeable Sephadex beads were higher than those obtained by a reference direct mass method, and some spore populations were separated into several density bands. With Percoll, all of the wet density values were about the same as those obtained by the direct mass method, and only single density bands resulted. The differences were due to the partial permeation of Metrizamide and Renografin, but not Percoll, into the spores and the permeable Sephadex beads. Consequently, the wet density of the entire spore was accurately represented only by the values obtained with the Percoll gradient and the direct mass method. The dry densities of the spores and particles were determined by gravity buoyant sedimentation in a gradient of two organic solvents, one of high and the other of low chemical density. All of the dry density values obtained by this method were about the same as those obtained by the direct mass method. PMID- 6285825 TI - Development of a method for detection of human rotavirus in water and sewage. AB - The simian rotavirus SA11 was used to develop a simple, reliable, and efficient method to concentrate rotavirus from tap water, treated sewage, and raw sewage by absorption to and elution from Filterite fiberglass-epoxy filters. SA11 adsorbed optimally to Filterite filters from water containing 0.5 mM AlCl3 at pH 3.5. Filter-bound virus was eluted with 0.05 M glycine-NaOH supplemented with 10% tryptose phosphate broth at pH 10. SA11 was quantitated by plaque assay, whereas human rotavirus was detected by immunofluorescence. The method was applied to detect rotavirus in raw and treated sewage at two Houston, Tex., sewage treatment plants. The sewage isolates were identified as rotavirus, probably a human strain, based on several criteria. The sewage isolates were detectable by an immunofluorescence test, using anti-SA11 serum which would detect the simian, human bovine, and porcine rotaviruses. No reaction was noted by immunofluorescence with the reoviruses or several common enteroviruses. The sewage isolates were neutralized by convalescent sera from a human adult and infant who had been infected by rotavirus as well as by a hyperimmune serum prepared in guinea pigs against purified human rotavirus. Preimmune or preillness sera did not react with the isolates by neutralization or immunofluorescence. The natural isolates were sensitive to pH 11 and other inactivating agents, similar to SA11. The buoyant density of the sewage isolates in CsCl gradients was 1.36 g/cm3, which is the value usually reported for complete, infectious rotavirus particles. The double-shelled particle diameter was 67.1 +/- 2.4 nm. Finally, electron micrographs of cell lysates inoculated with the sewage isolate showed particles displaying characteristic rotavirus morphology. PMID- 6285827 TI - Physical and kinetic properties of sheep liver pyridoxine kinase. PMID- 6285828 TI - DT-diaphorase as a quinone reductase: a cellular control device against semiquinone and superoxide radical formation. PMID- 6285829 TI - Nuclease I from suspension-cultured Nicotiana tabacum: purification and properties of the extracellular enzyme. PMID- 6285826 TI - An investigation into the role of hydroxyl radical in xanthine oxidase-dependent lipid peroxidation. PMID- 6285830 TI - A rapid procedure for the isolation and purification of photosynthetic reaction centers from Rhodopseudomonas sphaeroides R-26. PMID- 6285831 TI - Mechanism of action of gastric secretory inhibitors: effects of SCN- OCN-, NO-2, and NH+4 on (H+ + K+)-ATPase-mediated transport of H+ inside gastric microsomal vesicles. PMID- 6285833 TI - Rotational diffusion of cytochrome P-450 in the microsomal membrane-evidence for a clusterlike organization from saturation transfer electron paramagnetic resonance spectroscopy. PMID- 6285834 TI - The effect of chronic ethanol consumption on temperature-dependent physical properties of liver mitochondrial membranes. PMID- 6285832 TI - Irreversible inhibition of folate transport in Lactobacillus casei by covalent modification of the binding protein with carbodiimide-activated folate. PMID- 6285835 TI - Effects of nucleotides on activity of a purified ADP-ribosyltransferase from turkey erythrocytes. PMID- 6285836 TI - Tb3+ as a luminescent probe of actin structure: effects of polymerization, KI, and the binding of deoxyribonuclease I. PMID- 6285837 TI - Low-dose radioisotope scanning and quantitative analysis in the diagnosis of congenital hypothyroidism. AB - Quantitative thyroid scanning using low doses of 99mTc sodium pertechnetate (1.85 3.7 MBq) was performed in 38 cases of congenital hypothyroidism. Of these 38 cases, 29 were scanned at 14 +/- 6 days old, and 9 at 1 year old. The scans show the full range of gland anatomy from athyreotic to normal. All morphologically normal scans had grossly increased uptakes of 99mTc. The incidence of the various thyroid anatomies was different in each age group. The average radiation dose to the thyroid was 2.29 mGy, with at least 70% of patients receiving a dose of 3.0 mGy or less. Such low doses of 99mTc should allow further scanning in later life. Neonatal thyroid scanning reveals the aetiology of congenital hypothyroidism and enables the clinician to assess the short- and long-term needs of the child. PMID- 6285838 TI - [Non-functioning retroperitoneal paraganglioma. Report of a case]. PMID- 6285840 TI - [Multiple paraganglioma of the thyroid region of the neck. Papillary cancer associated with parathyroid adenoma (author's transl)]. PMID- 6285839 TI - [Fibrolamellar carcinoma of the liver. A case report (author's transl)]. PMID- 6285841 TI - [An outbreak of rotavirus gastroenteritis in a home for the elderly (author's transl)]. PMID- 6285842 TI - Upper extremity neuropathies following median sternotomy. AB - The status of 958 patients who underwent median sternotomy between January, 1978, and May, 1981, was analyzed. Fifty-four patients had an upper extremity neuropathy. Among 38 patients who underwent further evaluation, motor and sensory nerve conduction studies localized the injury to the level of the elbow in 13, to the brachial plexus in 10, and to both locations in 6. Ninety-two percent of these 38 patients were asymptomatic 3 months after operation. PMID- 6285843 TI - Inhibin interaction with LHRH receptors at the pituitary level. AB - In order to investigate whether inhibin could modulate the action of luteinizing hormone releasing hormone (LHRH), the in vitro effect of inhibin of LHRH bindings to the pituitaries from intact adult male rats was studied. The inhibin preparations suppressed the binding of labeled LHRH to pituitary receptors in a dose-related manner. In vivo administration of inhibin decreased the pituitary LHRH receptor concentration, with an apparent increase in the affinity of these receptors. A dose-related decrease was observed in serum follicle stimulating hormone (FSH) levels in the same group whereas the serum luteinizing hormone (LH) levels remained unaltered. There may be a direct action of inhibin at the pituitary level to suppress FSH levels specifically. PMID- 6285844 TI - Dose-dependent stimulation of antroduodenal coordination by domperidone via specific dopamine-receptors. PMID- 6285845 TI - Evaluation of the hypothalamic hypophyseal adrenal axis in patients receiving long-term hemodialysis. AB - Several alterations are present in the hypothalamic hypophyseal regulation of many hormones in patients with chronic renal failure. Evaluation of the hypothalamic hypophyseal adrenal axis in these groups of patients demonstrated normal levels of plasma cortisol. Dexamethasone suppression is abnormal after administration of 1 mg of oral dexamethasone, but normal after 3 mg. Dexamethasone blood levels were lower than the control after administration of 1 mg of oral dexamethasone. A dexamethasone metabolic clearance showed a similar half-life between the patients and controls. Oral absorption study showed poor absorption of the drug. Therefore, there is a problem of gastrointestinal absorption producing the abnormal dexamethasone suppression test in patients with renal failure. Results of metyrapone tests were normal. Corticotropin stimulation tests elicited a normal response. Insulin-induced hypoglycemia does not produce an increment in plasma cortisol or adrenocorticotropic hormone levels. PMID- 6285846 TI - Mutagenesis of Alcaligenes eutrophus by insertion of the drug-resistance transposon Tn5. AB - Drug-resistance element Tn5 coding for kanamycin resistance was used for mutagenesis of Alcaligenes eutrophus strain H16. The vehicle for introducing Tn5 into A. eutrophus was plasmid pJB4JI harboured by Escherichia coli. Kanamycin resistant transconjugants occurred at a frequency of approximately 5 x 10(-8). One third of the transconjugants exhibited other plasmid-coded resistances such as gentamycin and spectinomycin. However, the latter markers were not stably maintained in the new host. Among the kanamycin-resistant transconjugants three classes of mutants were found: (i) Auxotrophic mutants occurred at a frequency of 0.8% and showed requirements for histidine, methionine, aspartate or isoleucine. Out of eleven auxotrophic mutants examined eight reverted to prototrophy. However, none of the revertants was kanamycin-sensitive. (ii) Mutants unable to grow with fructose as the carbon source occurred at a frequency of almost 10%. (iii) Mutants which had lost the ability to grow autotrophically with hydrogen and carbon dioxide were found at a frequency of 1%. Further analyses revealed that this class of mutants was either defective in carbon dioxide fixation or impaired in hydrogen metabolism. PMID- 6285847 TI - Do cyanobacteria contain "mammalian-type" cytochrome oxidase? AB - The cytochrome content of membranes isolated from seven species of cyanobacteria was investigated in terms of conventional difference spectra, carbon monoxide difference spectra, photoaction spectra and photodissociation spectra, and by extraction of acid-labile heme followed by spectral identification. In addition, the effect of various inhibitors and activators on the oxidation of horse heart cytochrome c by the membrane was studied. Both the spectral features and the properties of the cytochrome oxidase reaction catalysed by the membranes suggested the presence of a terminal oxidase strikingly similar to mitochondrial ferrocytochrome c:oxygen oxidoreductase (EC. 1.9.3.1). PMID- 6285848 TI - Spontaneous rat nephroblastoma: ultrastructure of a transplant line. AB - Tumor tissue derived from a nephroblastoma arising spontaneously in an Nb hooded rat and carried for 56 generations as a subcutaneous syngeneic transplant was examined by electron microscopy. Histologically, the transplant showed the typical pattern of dense clusters or sheets of basophilic epithelioid cells set within ramifying tracts of fibrous mesenchyme. At the ultrastructural level, the tumor cells within dense clusters were arranged in anastomotic cords that circumscribed clear intervening spaces. The tumor cells were a monomorphic population possessing the morphologic characteristics of blast cells, including polyribosomes as the predominant cytoplasmic organelle. Furthermore, the close contiguity of cells within the cords, their polyhedral form, and their connection by intercellular junctions established the tumor cell type as epithelial. Consistent with benign stroma, mesenchymal tracts contained highly differentiated, mature fibroblasts, collagen, phagocytes, cells of the lymphoid series, and normal blood vessels. No transitional forms between epithelium and mesenchyme suggestive of bipotential differentiation were seen. Although the data represent only a single transplantation line, the observations support the contention (based on light microscopic appraisal of a number of primary tumors) that nephroblastoma in te rat can be a purely epithelial neoplasm. PMID- 6285849 TI - [A case of intraneural cystic ganglion of the external sciatico-popliteal nerve]. PMID- 6285850 TI - [Spinal deformity after surgery and radiotherapy of patients treated for Wilms' tumor, with an observation period of over 5 years]. PMID- 6285851 TI - [A case of ovarian arrhenoblastoma]. PMID- 6285853 TI - Endorphins in septic shock: hemodynamic and endocrine effects of an opiate receptor antagonist and agonist. AB - The pathophysiological role of endorphins in septic shock was studied in a porcine model. Septic shock was induced by the intravenous infusion of live Escherichia coli. Naloxone hydrochloride, an opiate receptor blocker, given during profound septic shock, increased blood concentrations of glucagon and cyclic adenosine monophosphate (cAMP), while BP and cardiac output increased transiently. Heart rate and hepatic glycogen value decreased, but insulin and cortisol levels remained unchanged. In contrast, exogenous morphine injection produced further reduction of BP, increased pulmonary wedge pressure, and increased substance P, while growth hormone level and cardiac output remained unchanged. Neither hormonal nor hemodynamic changes were noted in saline controls. Thus, the endogenous opiates appear partly responsible for the hemodynamic derangements during septic shock, and naloxone is able to reverse such depression, even though the effects are transient and relatively minor when naloxone is given late in the course of septic shock. Endogenous opiates also affect the hormonal homeostasis in shock, and there are indications that this may be mediated by the adenylate cyclase-cAMP system. PMID- 6285854 TI - Pertussis antigens--screening models on toxicity. AB - From the same batch of B. pertussis bacteria two types of DPT-vaccines were produced after harvest of the inactivated organisms by centrifugation or acid precipitation. The first vaccine contained whole pertussis bacteria and the second an extracted antigen complex. In a screening programme with reliable animal tests, such as a modified mouse-weight-gain assay, mouse leukocytosis test, histamine sensitization in mice, allergic encephalitis in rats, and the limulus test it could be demonstrated that the vaccine with whole inactivated bacteria was more toxic than that with extracted antigens. Cell material harvested by centrifugation showed a lower rate of toxicity than that containing acid-precipitated pertussis organisms. PMID- 6285855 TI - Toxicity testing in relation to aging. AB - Clinical evaluation of drugs is usually carried out in young adult subjects or patients and the data obtained is extrapolated to other drug users, e.g., the very old. Similarly, preclinical toxicity testing is invariably done in young animals. However, the main brunt of adverse drug reactions is borne by the elderly age group. Also many determinants of drug response - pathophysiology, pharmacokinetics and pharmacodynamics are different in the elderly. It is important to evaluate drug response both wanted and unwanted with this population in mind. Thus, it is appropriate that clinical effects be assessed in elderly populations if the drug is intended for use in this group. PMID- 6285852 TI - [Hormone receptors and cancer of the breast]. PMID- 6285856 TI - The induction of murine cytotoxic T lymphocytes by bluetongue virus. AB - After inoculation with live bluetongue virus, mice produced cytotoxic T lymphocytes (CTL) which showed virus and H-2 restriction. Inactivated preparations failed to induce CTLs. On secondary in vitro stimulation, specifically sensitised memory cells also produced high numbers of CTLs. The need for replicating virus to induce primary CTLs, evidence for partial type specificity and the role which cell-mediated immunity might play in the early stages of a bluetongue virus infection are discussed. PMID- 6285857 TI - Localization of foot-and-mouth disease--RNA synthesis on newly formed cellular smooth membranous vacuoles. AB - Viral RNA synthesis in foot-and-mouth disease infected bovine kidney cell cultures was associated throughout the infectious period with newly formed smooth membranous vacuoles. Membrane formation was measured by choline uptake. The site of RNA synthesis was determined by electron microscopic examination of autoradiograms of incorporated [3H] uridine. Both membrane formation and RNA synthesis became significant at 2.5 hours postinfection, but membrane formation increased steadily to 4.5 hours while RNA synthesis peaked at 3.5 hours. Percent density distributions of developed silver grains on autoradiograms showed that almost all RNA synthesis was concentrated on the smooth vacuoles of infected cells. Histogram analysis of grain density distributions established that the sites of RNA synthesis was the vacuolar membrane. The newly formed smooth membrane-bound vacuoles were not seen to coalesce into the large vacuolated areas typical of poliovirus cytopathogenicity. PMID- 6285858 TI - Virological diagnosis of enterovirus type 71 infections: experiences gained during an epidemic of acute CNS diseases in Hungary in 1978. PMID- 6285859 TI - Persistent asymptomatic infection of the laboratory mouse by simian foamy virus type 6: a new model of retrovirus latency. AB - Simian foamy virus (SFV) type 6, originally isolated from the kidney of a kuru inoculated chimpanzee, has been adapted to produce an asymptomatic infection in Swiss-Webster white mice, with virus detected in the kidney and spleen for up to 10 months after intra-peritoneal inoculation, and the presence in some animals of complement-fixing, but not neutralizing, serum antibody. This first successful experimental infection of the laboratory mouse by a representative of the foamy virus group has special interest as a convenient model in which to study pathogenesis and viral latency, particularly in view of the isolation and recent characterization of a foamy virus from human tissue which is virtually indistinguishable from the SFV type 6 used in the present study. PMID- 6285860 TI - Analysis of the TK enzyme complex induced by HSV types 1 and 2 by means of isoelectric focusing and polyacrylamide gel electrophoresis. AB - Recently we have described that the Herpes simplex virus (HSV)-induced thymidine kinase (TK) induces AMP- and ADP-dThd-5'-phosphotransferase activities. We now demonstrate the heterogeneity of the described activities in isoelectric focusing experiments and polyacrylamide gel electrophoresis. A TK--mutant of HSV type 1 fails to induce these activities. The activities of the type 1 enzyme complex was neutralized by an anti-HSV-serum. The TK-enzyme complex expressed in LTK--cells transformed to a TK+-phenotype by sheared HSV-1 DNA was compared with the wild type TK complex in isoelectric focusing experiments. Additionally we demonstrate that the HSV type 1 enzyme complex has thymidylate kinase activity, while the type 2 TK complex did not exhibit thymidylate kinase activity. Feedback regulation mechanisms by dTMP, dTDP and dTTP were investigated using partially purified enzyme preparations of HSV types 1 and 2 infected TK--cells. PMID- 6285861 TI - Isolation of lapine rotavirus in cell cultures. Brief report. AB - Two cytopathic rotavirus strains were isolated in MA 104 cells from diarrheal rabbits. The isolates showed marked cross reactions with strain Lincoln of bovine rotavirus in immunofluorescence, but no cross reaction in neutralization. PMID- 6285862 TI - Productive and abortive replication of human cytomegalovirus at different environmental pH values. PMID- 6285863 TI - Application of different strains for determining IgM antibodies against cytomegalovirus-induced late antigen. PMID- 6285864 TI - The virion RNA species of the Kirsten murine sarcoma-leukemia virus complex released from a clonally related series of mouse cells. AB - We have characterized the virion RNA species of Kirsten sarcoma (KiSV) and Kirsten leukemia (KiLV) viruses released from a clonally related series of mouse cells (14). We have identified the KiLV and KiSV genome RNAs. In addition to the viral RNA species we find large amounts of a virus-like RNA (VL30 RNA), which is heterogeneous and shows variability in its expression. The amount of VL30 RNA in virions does not correlate with the state of transformation of the cells releasing the virus or the ability of the virus to transform other cells. Characterization of RNA rescued from non-producer cells has revealed a sarcoma virus (KiSV CB3) with an oligonucleotide fingerprint different from that of a standard KiSV RNA, suggesting that it has lost some viral sequences. The oligonucleotide fingerprints of KiLV and VL30 RNAs are distinct from each other and from those reported for other murine leukemia virus RNAs. PMID- 6285865 TI - CV-1 cells release proteins that facilitate infection by simian virus 40 and echovirus 6. AB - Uninfected simian cells (CV-1) produce two different proteins, one of which enhances the infectivity of echovirus 6 and the other enhances the infectivity of SV40 in less susceptible cells. The enhancers are released by metabolizing cells into the culture medium. The SV40 enhancer protein is larger and less acidic than the echovirus enhancer. The SV40 enhancer protein can be dissociated into 2 subunits with apparent molecular weights of 42,000 and 24,000. The echovirus enhancer protein consists of 2 subunits with apparent molecular weights of 28,000 and 31,000. Electrophoretically purified enhancer proteins interact with virions and retain their biological activities and viral specificities. The SV40 enhancer stimulates expression of SV40-T antigen under conditions in which untreated virus does not initiate expression of T antigen. This result and the observation that the enhancers do not increase the infectivity of the nucleic acids of their respective viruses suggest that the enhancers act either at the stage of penetration or uncoating. PMID- 6285867 TI - The preparation of purified bluetongue virus group antigen for use as a diagnostic reagent. AB - Soluble group antigen isolated from bluetongue virus infected BHK cells has been shown to be a single protein with a molecular weight of 38,000 and to comigrate on polyacrylamide gels with protein 7 isolated from virus particles. Purification by exclusion chromatography and isoelectric focussing yields a protein that appears to be antigenicaly identical to the native group antigen found in virus harvests; it is non infectious for sheep and is thought to be largely free from host cell and other virus specified proteins. It is suggested that this antigen could be used in diagnostic procedures with safety. PMID- 6285866 TI - Reactivation in calves of Bovid herpesvirus 2 latent infection. AB - Seven calves which had recovered from an experimentally induced infection with Bovid herpesvirus 2 (BHV 2) were subjected to a course of dexamethasone (DMS) treatment six months later. BHV 2 was recovered from nasal swabs obtained from each calf 11 days after the start of DMS treatment. The virus was also isolated from the skin, nervous system tissue and lymph nodes of a calf killed 11 days after the start of DMS treatment in which a concurrent piroplasma infection was also, unexpectedly, discovered. It is suggested that the skin and nervous system are possible anatomical sites for the latency of BHV 2 in the host. PMID- 6285868 TI - Distribution of actin and myosin in a rat neuronal cell line infected with herpes simplex virus. AB - The indirect immunofluorescence technique was used to study alterations in the distribution of actin and myosin filaments in a rat B 103 neuronal cell line infected with herpes simplex virus type 1 (HSV-1). In uninfected cells, actin filaments were arranged in parallel and ran lengthwise from one end of the cell to the other; although myosin filaments were closely associated with actin filaments, additional myosin formed a netlike distribution which did not stain for actin. In infected cells, actin filaments became more randomly aligned and were concentrated along with myosin in close association with rosette-like formations of nuclei in syncytial cells; structural organization of actin and myosin within these intensely staining areas was no longer evident. The possible role of contractile proteins (actin and myosin) in viral infections of neural tissue is raised. PMID- 6285869 TI - The anatomical distribution of ADPase activity in the rabbit aorta. AB - The anatomical distribution of ADPase activity in the rabbit aorta was investigated. The aortic arch and upper thoracic regions of the rabbit aorta were found to have a reduced capacity to break down ADP and also unable to further metabolise the AMP thus formed. ADPase activity progressively increased down the aorta to the abdominal regions where it was highest. The abdominal regions of the aorta together with the lower thoracic region were able to produce adenosine from ADP. These results suggest a connection between ADPase activity and the incidence of atherosclerosis in rabbits. Thus in the aortic arch and upper thoracic regions of the aorta where the incidence of the disease is higher, the ability of the vascular tissue to break down ADP is low; therefore platelet aggregation is more likely to occur in response to minimal wear and tear. Conversely, in the abdominal regions where ADPase activity is highest and the incidence of the disease is lower ADPase may play a protective role in limiting ADP-induced thrombotic response to vascular trauma. PMID- 6285870 TI - [Ultracytochemistry of phosphohydrolases]. PMID- 6285871 TI - Oculomotor effects of intermittent conduction block in myasthenia gravis and Guillain-Barre syndrome. An oculographic study with computer simulations. AB - Five abnormal oculographic patterns were identified in eight patients with either myasthenia gravis or Guillain-Barre syndrome (GBS). These could be differentiated into three intrasaccadic and two postsaccadic abnormalities. From our studies of computer simulations, and considering the established pathophysiology of myasthenia gravis and GBS, we believe that our oculographic findings were a consequence of defects in peripheral neural and neuromuscular conduction, together with a simple adaptive increase in duration of the saccadic burst of central innervation. We conclude that the eye movement abnormalities we observed are explained by intermittent block of peripheral conduction, and suggest that any disease causing intermittent blockage of neural signals to extraocular muscles will produce similar abnormalities of eye movement. PMID- 6285872 TI - Effects of timoptic on corneal reepithelialization. AB - The effects of topical 0.5% Timoptic on the reepithelialization of rabbit corneas were compared to the effects observed in rabbits receiving either vehicle, phosphate-buffered saline (PBS), PBS containing 0.5% timolol maleate, or no treatment. The rate of epithelial wound closure was slowed, and epithelial DNA content was less in both Timoptic- and vehicle-treated corneas. Light microscopy showed that the epithelium of animals treated with either Timoptic or vehicle processed fewer cellular layers in the wounded areas as well as considerable disruption of the epithelial layer. The rate of DNA synthesis and the cyclic adenosine monophosphate content of the epithelial cells from all treatment groups were statistically the same. The impaired wound closure observed during Timoptic and vehicle treatment was probably due to the sloughing of cells caused by the 0.01% benzalkonium chloride in the vehicle and not to impaired healing caused by the metabolic effects of timolol meleate. PMID- 6285873 TI - Metastasis to the choroid complicating adenoid cystic carcinoma. AB - Two months after a complete response to doxorubicin hydrochloride and cyclophosphamide administered for systemic effects, with the disappearance of pulmonary and pleural metastases from adenoid cystic carcinoma of salivary origin, a patient experienced symptomatic choroidal metastasis with no other evidence of active disease. Irradiation produced complete disappearance of the choroidal metastasis and restoration of visual acuity. The literature is reviewed, and the unusual nature of this metastasis is discussed. PMID- 6285874 TI - Small cell epidermoid carcinoma of salivary glands. 'Pseudo'-oat cell carcinoma. AB - Two patients had small cell carcinomas of the salivary glands, with pathological features indicating squamous differentiation, heretofore not described. One is free of disease at seven years, and the second is alive, with regional metastases at four years. Sections from one tumor were studied by electron microscopy and revealed tonofilaments and desmosomes. Most cases of small cell carcinomas of the salivary glands have been considered akin to bronchogenic oat cell carcinoma. Their less aggressive behavior, however, suggests that at least some of these tumors were not true oat cell carcinomas. Our findings, and those of others, indicate that small cell carcinomas of the salivary glands (or head and neck) represent a heterogeneous group. Electron microscopy should be used to determine the exact nature of these neoplasms. If an oat cell nature is ruled out, local and regional treatment should be aggressive, since small cell carcinomas other than oat cell appear not to have a dismal prognosis. PMID- 6285875 TI - Synovial cell sarcoma of the temporomandibular joint. Computed tomographic findings. PMID- 6285876 TI - Cyclic AMP and gastric acid secretion in the dog. AB - Canine gastric secretions were collected for acid and cyclic AMP assay at zero, 15, 30, 45 and 60 minutes during intravenous infusion of zero, 2.5 and 5.0 micrograms/kg/hr of pentagastrin. A resulting brisk acid response was not accompanied by an elevation of cyclic AMP content. It is thereby suggested that cyclic AMP may not be an obligatory intermediary for gastric acid secretion. PMID- 6285877 TI - The early pathogenesis of turkey herpesvirus infection in chickens and turkeys. AB - The early responses of chickens and turkeys to infection with two strains of turkey herpesvirus (FC 126 and a recent field isolate, T-5A) were compared. All birds had similar virus rescue patterns from the spleen, regardless of virus strain used for infection. Productive-restrictive infections as indicated by the presence of viral antigens were found only in the spleens of infected turkeys. Viral antigens were not found in bursa or thymus but were found intermittently and at low levels in the skin. Small focal accumulations of lymphocytic cells were found in the nerves of all groups of birds 7 and 14 days postinoculation. PMID- 6285878 TI - Effect of endogenous leukosis virus genes on response to infection with avian leukosis and reticuloendotheliosis viruses. AB - We examined the effect of the presence or absence of endogenous viral gene (ev) 3, which controls expression of group-specific viral and envelope antigens (gs+chf+ phenotype), and ev2, which controls the production of a complete subgroup E virus (V-E+ phenotype), on the response of chickens to RAV-1, an exogenous avian leukosis virus (ALV) with an antigenic relationship to endogenous virus. After inoculation at one day of age, the chickens lacking either ev gene expression had a lower frequency of virus isolations and higher frequency and titer of neutralizing antibodies than those expressing ev genes. This relationship was not seen in groups inoculated with chick syncytial virus (CSV), a reticuloendotheliosis-associated virus with no relationship to endogenous virus, but the ev2+ birds tended to yield more CSV isolations than the ev2- birds. We suggest that chickens expressing ev genes may be immunologically tolerant to antigens common to exogenous and endogenous viruses. In addition, ev3 birds inoculated with RAV-1 at one day of age or as embryos died at a high rate between 6 and 12 weeks of age with a non-neoplastic syndrome characterized by severe atrophy of lymphoid organs, an inflammatory reaction in the liver, and a lower immune response to particulate antigens. PMID- 6285879 TI - Infectivity for chicken embryos of tissue-culture-modified infectious laryngotracheitis virus. AB - The present study shows the infectivity for chicken embryos of the SPL strain of tissue-culture-modified infectious laryngotracheitis (ILT) virus. The modified virus formed small pocks on the chorioallantoic membrane (CAM) and gave no adverse reaction after administration to chickens. For 1- and 6-day-old embryonated specific-pathogen-free (SPF) eggs, the SPL strain showed higher infectivity than virulent strain NS-175 or wild strains of ILT virus. A high percentage of embryos of day-old eggs inoculated with strains SPL via the yolk sac died within several days, and virus was recovered from yolk and embryos with high titer. In the embryos inoculated at 6 days of age, histopathological lesions characterized by syncytium formation with intranuclear inclusion bodies were observed mainly in the CAM, liver cells, epithelial cells of the oral cavity, and esophagus and rarely in epithelial cells of the uriniferous tubule, bronchus, trachea, choroid plexus, or feather follicles. In the 10-day-old embryonated SPF eggs inoculated via the chorioallantoic cavity, however, the SPL strain demonstrated low pathogenicity compared with strain NS-175 and the two wild strains. PMID- 6285880 TI - Dose-dependent inhibition of virus rescue from lymphocytes latently infected with turkey herpesvirus or Marek's disease virus. AB - The number of plaque-forming units (PFU) of turkey herpesvirus (HVT) isolated per 10(6) latently infected splenic lymphocytes was determined by co-cultivation on permissive monolayer cultures in 35-mm-diameter Petri dishes. Doses of 1 x 10(6) spleen cells or less per culture gave uniform dose-related titers, whereas doses of 8 x 10(6) cells often yielded less than 1-2% of the expected number of PFU. Intermediate doses gave proportionally reduced virus yields. This dose-dependent inhibition was observed with spleen cells from birds within a week after infection and became more marked with time. A similar phenomenon occurred with a non-oncogenic Marek's disease virus (MDV) isolate (SB-1) but not with oncogenic MDV isolates (CU-2, JM-10, GA-5), except in genetically resistant birds. High numbers of uninfected spleen cells mixed with low numbers of HVT-infected cells during assay reduced titers only slightly. Immunosuppression by combined neonatal thymectomy and cyclophosphamide treatment before HVT infection prevented the inhibition, but embryonal bursectomy had no effect. PMID- 6285881 TI - The effect of incubation temperature on the propagation of duck adenovirus (virus 127-like) in duck and chicken cells. AB - Duck adenovirus (Cornell strain) was propagated in duck and chicken embryo cells at 37.5 C and at 40 C. In duck cells, virus levels, as indicated by HA titers, peaked earlier at 40 C than at 37.5 C. High titers were eventually observed in duck cells at both temperatures. In chicken embryo fibroblasts, no titers were observed at 37.5 C, whereas low titers were observed at 40 C. Evidence of virus propagation was not detected in chicken embryo liver and kidney cells. PMID- 6285882 TI - Infectious bursal disease virus and Alcaligenes faecalis infections in turkeys. AB - To determine if infectious bursal disease virus (IBDV) augments alcaligenes rhinotracheitis (ART), turkey poults were exposed to IBDV, Alcaligenes faecalis, or both IBDV and A. faecalis. In five experiments, poults exposed to IBDV alone exhibited neither signs of disease nor histopathologic lesions. Serum antibodies to IBDV were detected in poults exposed to this virus by inoculation and by direct contact with inoculated birds. Signs of ART were observed 4 to 6 days following exposure to A. faecalis. Clinical signs of ART and histopathologic lesions in the upper respiratory tract of poults exposed to both IBDV and A. faecalis were similar to those observed in poults exposed to A. faecalis alone. PMID- 6285883 TI - Infectious laryngotracheitis in peafowl and pheasants. PMID- 6285886 TI - Symposium on: recent developments concerning the interdisciplinary research on biomembranes. PMID- 6285884 TI - Psittacine pox virus: virus isolation and identification, transmission, and cross challenge studies in parrots and chickens. AB - An avian pox virus was isolated from Amazon parrots dying with severe diphtheritic oral, esophageal, and crop lesions. The virus was propagated on chorioallantoic membranes (CAM) of 10-day-old chicken embryos, and a homogenate of the infected CAM was rubbed vigorously onto the conjunctiva, oral mucosa, and defeathered follicles of two healthy Amazon parrots and three conures. All experimental birds developed cutaneous and ocular pox lesions, and one parrot developed oral pox lesions. Specific-pathogen-free chicks inoculated with the virus isolate developed skin lesions identical to those of the parrots. Chickens vaccinated with fowl and pigeon pox vaccines and inoculated with the psittacine isolate developed lesions typical of avian pox. Chickens vaccinated with the psittacine virus were susceptible to fowl and pigeon pox virus infection. This pox virus isolate may thus be regarded as a potential pathogen for chickens. PMID- 6285885 TI - Pharmacological generality of the Avfail effect. PMID- 6285887 TI - Concomitant appearance of antibodies to two viruses in cerebrospinal fluid of an infant with acute encephalitis. AB - Acute encephalitis developed in a 5 1/2-month-old infant with a previous history of uncomplicated measles at 4 months of age. The patient survived his acute illness with resultant severe neurological sequelae. The serum complement fixing antibody titer for herpes simplex virus in the early phase of the illness was less than 1:4, and increased significantly to 1:64 in the convalescent phase. Antibody was also demonstrated in the cerebrospinal fluid. Antimeasles antibody was found in the cerebrospinal fluid in the early convalescent stage and remained at a detectable level for at least 1 month. Moreover, at least 71% of the total protein in the cerebrospinal fluid was IgG. Although brain biopsy for viral isolation was not performed, the most probable diagnosis was herpes simplex virus encephalitis, and the concomitant appearance of measles antibody in the cerebrospinal fluid may have been due to leakage of measles antibody from the blood into the cerebrospinal fluid due to disturbance of the blood-brain barrier. PMID- 6285888 TI - Purification of total cellular DNA from a single plant. AB - A procedure is outlined for purifying DNA from a single plant. A crude organelle pellet consisting of nuclei, chromatin, chloroplasts, and mitochondria is prepared, suspended, and immediately lysed with detergents. The DNA is separated from RNA, protein, and polysaccharides by banding it in CsCl density equilibrium gradients. Ethidium bromide is included in all buffers to act as an inhibitor of DNAase activity. The DNA prepared in this manner can be digested with restriction endonucleases, separated by gel electrophoresis, and used to identify specific genes by hybridization of cloned DNA sequences. PMID- 6285889 TI - Threonine accumulation in the seeds of a barley mutant with an altered aspartate kinase. AB - Barley (Hordeum vulgare L.) mutants altered in the regulation of synthesis of aspartate-derived amino acids were sought by screening embryos for growth on a medium containing lysine plus threonine. One mutant, Rothamsted 2501, was selected with good growth. From the segregation of resistance in the following generations, it was concluded that the resistance was conferred by a dominant gene, Lt1. No homozygous Lt1/Lt1 fertile plants have been recovered. Partially purified aspartate kinase preparations from resistant and sensitive plants were separated on DEAE-cellulose chromatography into three peaks of activity (I, II, III) and the feedback regulatory properties of these peaks determined. These peaks are considered to be three isozymic forms of aspartate kinase, one predominantly sensitive to threonine and two sensitive to lysine or lysine plus S adenosyl methionine. The feedback characteristics of one of the peaks of aspartate kinase activity from resistant plants were changed such that lysine was half-maximally inhibitory at 10 rather than 0.4 mM. Increases in te concentrations of the free pools of threonine (4x) and methionine (2x) were measured in young plants grown on a basal medium. Threonine in the soluble fraction of mature seeds from resistant plants was increased from 0.8 to 9.6% of the total threonine content. The total content of both threonine and methionine of the seeds was increased by 6% compared with grain of similar nitrogen content. PMID- 6285890 TI - Reactions of Adriamycin with haemoglobin. Superoxide dismutase indirectly inhibits reactions of the Adriamycin semiquinone. AB - The Adriamycin semiquinone produced by the reaction of xanthine oxidase and xanthine with Adriamycin has been shown to reduce both methaemoglobin and cytochrome c. In air, but not N2, both reactions were inhibited by superoxide dismutase. With cytochrome c, superoxide formed by the rapid reaction of the semiquinone with O2, was responsible for the reduction. However, even in air, methaemoglobin was reduced directly by the Adriamycin semiquinone. Superoxide dismutase inhibited this reaction by removing superoxide and hence the semiquinone by displacing the equilibrium: Semiquinone + O2 in equilibrium or formed from quinone + O2-. to the right. This ability to inhibit indirectly reactions of the semiquinone could have wider implications for the protection given by superoxide dismutase against the cytotoxicity of Adriamycin. Oxidation of haemoglobin by Adriamycin has been shown to be initiated by a reversible reaction between the drug and oxyhaemoglobin, producing methaemoglobin and the Adriamycin semiquinone. Reaction of the semiquinone with O2 gives superoxide and H2O2, which can also react with haemoglobin. Catalase, by preventing this reaction of H2O2, inhibits oxidation of oxyhaemoglobin. Superoxide dismutase, however, accelerates oxidation, by inhibiting the reaction of the semiquinone with methaemoglobin by the mechanism described above. Although superoxide dismutase has a detrimental effect on haemoglobin oxidation, it may protect the red cell against more damaging reactions of the Adriamycin semiquinone. PMID- 6285891 TI - The inositol trisphosphate phosphomonoesterase of the human erythrocyte membrane. AB - Human erythrocyte ghosts exhibit an inositol trisphosphate phosphomonoesterase activity that rapidly converts inositol 1,4,5-trisphosphate into inositol 1,4 bisphosphate and Pi. Degradation of the released inositol 1,4-bisphosphate is not observed. This activity is dependent on Mg2+ (or Mn2+) and it is not activated by Ca2+. Optimum activity is around pH 7 and activity is abolished by heat denaturation. The Km for inositol trisphosphate is approx. 25 microM. 2,3 bisphosphoglycerate is a competitive inhibitor, with a Ki of approx. 0.35 mM. Glycerophosphoinositol 4,5-bisphosphate is attacked at about one-eighth of the rate for inositol trisphosphate, but glycerophosphoinositol 4-phosphate is not a substrate. Incubation of 32P-labelled erythrocyte membranes with Mg2+ causes little breakdown of phosphatidylinositol 4,5-bisphosphate, the parent compound from which both glycerophosphoinositol 4,5-bisphosphate and inositol 1,4,5 trisphosphate are derived. On the basis of its substrate specificity and the inhibition by 2,3-bisphosphoglycerate, we suggest that this enzyme is selective for the 5-phosphate in those water-soluble phosphate esters of inositol that possess the vicinal pair of 4,5-phosphates but that it may also interact less strongly with other water-soluble compounds that have pairs of vicinal phosphates. PMID- 6285892 TI - Structural features of the first component of human complement, C1, as revealed by surface iodination. AB - Lactoperoxidase-catalysed surface iodination and sucrose-gradient ultracentrifugation were used to investigate the structure of human complement component C1. 1. Proenzymic subcomponents C1r and C1s associated to form a trimeric C1r2-C1s complex (7.6 S) in the presence of EDTA, and a tetrameric Clr2 C1s2 complex (9.1 S) in the presence of Ca2+. Iodination of the 9.1 S complex led to a predominant labelling of C1r (70%) over C1s (30%), essentially located in the b-chain moiety of C1r and in the a-chain moiety of C1s. 2. Reconstruction of proenzymic soluble C1 (15.2 S) from C1q, C1r and C1s was partially inhibited when C1s labelled in its monomeric form was used and almost abolished when iodinated C1r was used. Reconstruction of fully activated C1 was not possible, whereas hybrid C1q-C1r2-C1s2 complex was obtained. 3. Iodination of proenzymic or activated C1 bound to IgG-ovalbumin aggregates led to an equal distribution of the radioactivity between C1q and C1r2-C1s2. With regard to C1q, the label distribution between the three chains was similar whether C1 was in its proenzymic or activated form. Label distribution in the C1r2-C1s2 moiety of C1 was the same as that obtained for isolated C1r2-C1s2, and this was also true for the corresponding activated components. However, two different labelling patterns were found, corresponding to the proenzyme and the activated states. PMID- 6285894 TI - Solubilization and purification of rat liver 5'-nucleotidase by use of a zwitterionic detergent and a monoclonal-antibody immunoadsorbent. AB - 1. A variety of detergents were used to solubilize 5'-nucleotidase from rat liver plasma membranes. 2. The zwitterionic detergent Sulphobetaine 14 gave optimal solubilization by the criteria of release into a high-speed-centrifugation supernatant and the formation of the smallest and least polydisperse active enzyme observed on polyacrylamide-gel electrophoresis. 3. The Sulphobetaine 14 solubilized enzyme from rat liver was purified by using the conventional techniques of ion-exchange chromatography and gel filtration, or by an immunoaffinity step with a monoclonal antibody immunoadsorbent. 4. 5' Nucleotidase was purified at least 12 000-fold relative to liver homogenate by the immunoaffinity purification scheme and had a specific activity in the range 285-340 mumol/min per mg of protein. The yield was in the range 9-16%. 5. The purified enzyme shows a major polypeptide band of apparent Mr 70 000 on sodium dodecyl sulphate/polyacrylamide-gel electrophoresis and a minor band of apparent Mr 38 000. 6. A rational approach to the general problem of the purification of minor intrinsic membrane proteins is discussed, with the use of polyacrylamide gel electrophoresis to determine the most appropriate detergent and monoclonal antibodies in subsequent immunoaffinity purification. PMID- 6285893 TI - Partial purification of collagenase and gelatinase from human polymorphonuclear leucocytes. Analysis of their actions on soluble and insoluble collagens. AB - The separation and further purification of human polymorphonuclear-leucocyte collagenase and gelatinase, using modifications of the method of Cawston & Tyler [(1979) Biochem J. 183, 647-656], are described. The final preparations yielded collagenase of specific activity 260 units/mg and gelatinase of specific activity 13 000 units/mg. Gelatinase was purified to apparent homogeneity in a latent form, and analysis of the activation of 125I-labelled latent enzyme by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis and gel-filtration techniques suggested that no peptide material was lost on conversion into the active form. The purified natural inhibitors alpha 2-macroglobulin, tissue inhibitor of metalloproteinases ('TIMP') and amniotic-fluid inhibitor of metalloproteinases all inhibited the two polymorphonuclear-leucocyte metalloproteinases, but the last two inhibitors were slow to act and complete inhibition was difficult to attain. Collagenase degraded soluble types I and III collagen equally efficiently, but soluble type II collagen less well. Gelatinase alone had little activity on these substrates, although it enhanced the action of collagenase. Gelatinase was capable of degrading soluble types IV and V collagen at 25 degrees C, whereas collagenase was only active at higher temperatures when the collagens were susceptible to trypsin activity. By using tissue preparations of insoluble collagens (type I, II or IV) the activity of leucocyte collagenase was low and gelatinase activity was negligible, as measured by the solubilization of hydroxyproline-containing material. The two enzymes together were two or three times more effective in the degradation of these insoluble collagens. PMID- 6285895 TI - Studies by electron-paramagnetic-resonance spectroscopy of the molybdenum centre of aldehyde oxidase. AB - Molybdenum(V) e.p.r. spectra from reduced forms of aldehyde oxidase were obtained and compared with those from xanthine oxidase. Inhibited and Desulpho Inhibited signals from aldehyde oxidase were fully characterized, and parameters were obtained with the help of computer simulations. These differ slightly but significantly from the corresponding parameters for the xanthine oxidase signals. Rapid type 1 and type 2 and Slow signals were obtained from aldehyde oxidase, but were not fully characterized. From the general similarities of the signals from the two enzymes, it is concluded that the ligands of molybdenum must be identical and that the overall co-ordination geometries must be closely similar in the enzymes. The striking differences in substrate specificity must relate primarily to structural differences in a part of the active centre concerned with substrate binding and not involving the catalytically important molybdenum site. PMID- 6285896 TI - A method for determining kinetic parameters at high enzyme concentrations. AB - A graphical method is described which allows determination of kinetic parameters when substrate, inhibitor or activator concentrations must be in the vicinity of the enzyme concentration and a significant fraction of ligand is bound. Velocity is measured at several ligand: enzyme ratios at two or more enzyme concentrations. Results are obtained in terms of free and bound ligand corresponding to particular velocities. The relationship between velocity and bound and free ligand may then be analysed by any desired plotting technique. Preknowledge of the reaction mechanism or experimental determination of Vmax. is not required. The relationship between ligand bound and enzyme activity need not be linear and the method is equally suitable for analysing co-operative as well as simple kinetics. Application of the method is demonstrated by analysis of the inhibition of fructose, 1,6-bisphosphatase by AMP. PMID- 6285897 TI - Odorous chemical perturbations of (Na+ + K+)-dependent ATPase activities. Effects on native and lipid-substituted preparations from individual turbinals from dog olfactory tissue. AB - Individual turbinals from the right and left sides of dog olfactory tissue were removed and nerve-ending-particle preparations were prepared. (Na+ + K+) dependent ATPase activities of the individual preparations, and the effect of several odorous compounds [including (+)- and (-)-carvone] on the (Na+ + K+) dependent ATPase activities, were determined. The maximally stimulatory odorant concentration in the reaction mixture for the majority of odorants was found to be 1.0 mM. Matched pairs of left/right turbinals showed a lack of bilateral symmetry of response. (Na+ + K+)-dependent ATPase activities of various dog brain nerve-ending particle preparations responded only slightly to 1.0 mM odorants. The role of phospholipids in the (Na+ + K+)-dependent ATPase activity was found to be critical. Partial replacement of endogenous lipid with either synthetic phospholipids or extracted lipids resulted in changes in stimulation obtained with endogenous lipids alone. PMID- 6285898 TI - The SalGI restriction endonuclease. Purification and properties. AB - The type II restriction endonuclease SalGI has been purified to near homogeneity. At least 80% of the protein remaining after the final stage of the preparation is SalGI restriction endonuclease; no contaminating nucleases remain detectable. The principal form of the protein under both native and denaturing conditions is a monomer of M(r) about 29000. The optimal conditions for both enzyme stability and enzyme activity have been determined. PMID- 6285899 TI - The SalGI restriction endonuclease. Mechanism of DNA cleavage. AB - The cleavage of supercoiled DNA of plasmid pMB9 by restriction endonuclease SalGI has been studied. Under the optimal conditions for this reaction, the only product is the linear form of the DNA, in which both strands of the duplex have been cleaved at the SalGI recognition site. DNA molecules cleaved in one strand at this site were found to be poor substrates for the SalGI enzyme. Thus, both strands of the DNA appear to be cleaved in a concerted reaction. However, under other conditions, the enzyme cleaves either one or both strands of the DNA; the supercoiled substrate is then converted to either open-circle or linear forms, the two being produced simultaneously rather than consecutively. We propose a mechanism for the SalGI restriction endonuclease which accounts for the reactions of this enzyme under both optimal and other conditions. These reactions were unaffected by the tertiary structure of the DNA. PMID- 6285900 TI - The SalGI restriction endonuclease. Enzyme specificity. AB - We have analysed the kinetics of DNA cleavage in the reaction between the SalGI restriction endonuclease and plasmid pMB9. This reaction is subject to competitive inhibition by DNA sequences outside the SalGI recognition site; we have determined the Km and Vmax. for the reaction of this enzyme at its recognition site and the KI for its interaction at other DNA sequences. We conclude that the specificity of DNA cleavage by the enzyme is only partly determined by the discrimination it shows for binding at its recognition sequence compared with binding to other DNA sequences. PMID- 6285901 TI - Nucleotide sequence of the genes for beta and epsilon subunits of proton translocating ATPase from Escherichia coli. PMID- 6285902 TI - Activation of the virion-associated RNA polymerase of vesicular stomatitis virus by melittin. PMID- 6285903 TI - A collagen-like glycoprotein from elastin-rich tissues. PMID- 6285904 TI - Linked transcripts of the genes for leader and N message are synthesized in vitro by vesicular stomatitis virus. PMID- 6285905 TI - Gangliosides do not raise cyclic AMP levels during inhibition of lymphocyte mitogenesis. PMID- 6285906 TI - Changes in cyclic nucleotide levels correlated with growth, division, and morphology in Chlamydomonas chemostat culture. PMID- 6285907 TI - Internalization of beta-adrenergic receptor binding sites: involvements of lysosomal enzymes. PMID- 6285909 TI - A ganglioside receptor on lymphocytes mediates recognition of self. PMID- 6285908 TI - beta-Endorphin inhibits met-enkephalin breakdown by a brain aminopeptidase: structure-activity relationships. PMID- 6285910 TI - The effect of oxygen on fructose-2, 6-bisphosphate concentration and fructose-6 phosphate, 2-kinase activity in yeast cells. PMID- 6285911 TI - Glucocorticoids down-regulate the number of 1, 25-dihydroxyvitamin D3 receptors in mouse intestine. PMID- 6285912 TI - Photoaffinity labeling of beta-adrenergic receptors: identification of the beta receptor binding site(s) from turkey, pigeon, and frog erythrocyte. PMID- 6285913 TI - Cytochalasin E diminishes the lag phase in the release of superoxide by human neutrophils. PMID- 6285914 TI - Recombinant DNA-derived methionyl human growth hormone is similar in membrane binding properties to human pituitary growth hormone. PMID- 6285915 TI - Stimulatory effects of prolactin and anti-prolactin receptor serum on prolactin binding sites in rat liver cells in suspension culture. PMID- 6285916 TI - Hydrolysis of phosphoric acid diesters by snake venom phosphodiesterase and 5' nucleotide diesters of sinapis alba germs via and covalently enzyme-bound intermediate. PMID- 6285917 TI - In vitro maturation of ovine fetal adrenal cells adenylate cyclase: corticotropin dependent and independent development of the response to corticotropin. PMID- 6285918 TI - The conversion of leukotriene C4 to isomers of leukotriene B4 by human eosinophil peroxidase. PMID- 6285919 TI - Inhibition of protein synthesis on homologous and heterologous desensitization in the rat adipocyte. PMID- 6285920 TI - Interaction of manganese (II) with valinomycin: observation of mixed complexes. PMID- 6285921 TI - The oligopeptide chemotactic factor receptor on human polymorphonuclear leukocyte membranes exists in two affinity states. PMID- 6285922 TI - [D-Pen2, L-Cys5]enkephalinamide and[D-Pen2, D-Cys5] enkephalinamide, conformationally constrained cyclic enkephalinamide analogs with delta receptor specificity. PMID- 6285923 TI - Isolation of the GDP binding protein from brown adipose tissue mitochondria of several animals and amino acid composition study in rat. PMID- 6285925 TI - Aniline is hydroxylated by the cytochrome P-450-dependent hydroxyl radical mediated oxygenation mechanism. PMID- 6285924 TI - The presence of redox-sensitive nickel in the periplasmic hydrogenase from Desulfovibrio gigas. PMID- 6285926 TI - The influence of Mg2+ on the phosphorylation and dephosphorylation of myosin by an actomyosin preparation from vascular smooth muscle. PMID- 6285927 TI - Altered distribution and properties of cAMP-dependent protein kinase isozymes in spontaneously hypertensive rat aorta. AB - cAMP-dependent protein kinase activity was reduced in the cytosol fraction of spontaneously hypertensive rat (SHR) aorta compared to that of the Kyoto Wistar control rat (WKY). Two major peaks, isozymes I and II, of soluble cyclic AMP dependent protein kinase activity could be separated by DEAE-cellulose chromatography. The distributions of isozymes I and II were 40 and 60%, respectively, in WKY compared to 26 and 74% in SHR. Isozyme I of SHR eluted at a conductance of 2-3 mmhos compared to 5-6 mmhos in WKY. In addition, activity under the peak of isozyme I of SHR was reduced by approximately 55% compared to WKY. The half-life of thermal denaturation of isozyme I at 50 degrees was 21 min in WKY compared to 84 min in SHR. On the other hand, for isozyme II no significant differences were observed between WKY and SHR in elution pattern, total activity under the peak, or thermal denaturation of enzyme activity. These data suggest that specific changes had occurred in isozyme I of SHR. PMID- 6285929 TI - Inactivation of angiotensin II receptors in bovine adrenal cortex by dithiothreitol: further evidence for the essential nature of disulfide bonds. AB - Dithiothreitol (DTT), a disulfide reducing agent, significantly decreased specific (( 3H]angiotensin II binding to membrane-bound and solubilized bovine adrenal cortical receptors. Scatchard analysis indicated a reduction in the maximum number of membrane-bound binding sites without change in affinity. The effect of DTT on membrane-bound receptors was readily reversed by the sulfhydryl oxidizing agent, 5,5'-dithiobis-(2-nitrobenzoic acid) (DTNB), indicating that its action upon specific[3H]angiotensin II binding is mediated via reduction of disulfide bonds rather than other properties of this agent. Preincubation of the membranes with unlabeled angiotensin II, but not its biologically inactive precursor angiotensin I, protected the receptor from deactivation by DTT. The data provide evidence that disulfide bonds essential for[3H]angiotensin II receptor binding in bovine adrenal cortex are located at, or near, the active sites of the receptor. PMID- 6285928 TI - Maternal methadone administration: deficit in development of alpha-noradrenergic responses in developing rat brain as assessed by norepinephrine stimulation of 33Pi incorporation into phospholipids in vivo. AB - The effects of perinatal methadone exposure on the development of noradrenergic responses in the brain were examined by assessing the ability of intracisternally administered norepinephrine to stimulate 33Pi incorporation into phospholipids in vivo; the effect of norepinephrine is mediated by alpha1-receptors juxtaposed to noradrenergic nerve terminals. Although there was no difference in basal (unstimulated) incorporation of 33Pi, a deficit in norepinephrine-induced stimulation of incorporation was found throughout the preweaning period in offspring of dams treated daily with methadone beginning in midgestation. This effect was not seen when methadone was given during the postnatal period. Since perinatal methadone exposure also delays development of presynaptic catecholaminergic nerve terminals in the brain, these results support the view that perinatal exposure to methadone depresses overall central noradrenergic synaptic function; however, the effects on presynaptic development and on receptor-mediated responses appear to be separable in that they display differences in the critical age periods of sensitivity to perturbation by the drug. PMID- 6285930 TI - Effects of propranolol and a number of its analogues on sodium channels. AB - To assess the relative contributions that the sodium channel blocking activity of propranolol may play in a variety of its therapeutic applications, its effects were examined in vitro with a sodium channel specific 22Na+ uptake system, using rat brain membranes. Propranolol inhibited 22Na+ uptake in the rat brain membrane preparation by acting as a competitive inhibitor of the binding of the sodium channel opening agent veratridine, with an IC50 for this action of 6.5 microM. This is approximately one order of magnitude higher in concentration than that necessary for expression of the beta-adrenergic antagonism of propranolol. The binding of propranolol and its action to block sodium channels were demonstrably different from those of the neurotoxins tetrodotoxin and saxitoxin. Propranolol had effects on sodium channels that are similar, although not identical to those of the local anesthetics procaine and lidocaine. The concentrations of propranolol and a number of its analogues which produced 50% inhibition of 22Na+ uptake (IC50 values ranging from 4 to greater than 100 microM) were similar to the concentrations of these same analogues which were required to produce negative inotropic and antiarrythmic effects (ED40) on isolated rabbit atria [D. O. Rauls and J. K. Baker, J. med. Chem, 22, 81 (1979)]. These effects showed correlations of 0.945 and 0.936, respectively, with the 22Na+ uptake inhibition. It is concluded from this information that a substantial proportion of the negative inotropic and antiarrythmic effects of propranolol is due to its action on sodium channels. PMID- 6285931 TI - Regulation of phosphoribosylpyrophosphate synthetase by endogenous purine and pyrimidine compounds and synthetic analogs in normal and leukemic white blood cells. AB - Phosphoribosylpyrophosphate (PRPP) is essential for the formation of both purine and pyrimidine nucleotides as well as for the active nucleotide form of some chemotherapeutic agents. The formation of PRPP is catalyzed by by enzyme PRPP synthetase, and many different compounds are known to affect the activity of this enzyme. This report examines the effects of endogenous purine and pyrimidine nucleotides, nucleosides, and several analogs of these compounds on the activity of PRPP synthetase from different types of normal and leukemic white blood cells (i.e. normal lymphocytes, normal granulocytes, phytohemagglutinin-stimulated lymphocytes, and acute and chronic leukemic cells). Our results show that the effect varied with each individual compound, and the magnitude of the effect was dependent on the source of the enzyme. Since it appears possible to differentially affect PRPP synthetase activity from the different types of leukemic cells, this enzyme may be a potential target site in the chemotherapy of leukemia. PMID- 6285934 TI - Interleukin-2 deficiency, genes, and systemic lupus erythematosus. PMID- 6285933 TI - Binding of depolarizing drugs to the ionic channel sites of the nicotinic acetylcholine receptor. PMID- 6285935 TI - Studies regarding a possible function for viruses in the pathogenesis of systemic lupus erythematosus. PMID- 6285936 TI - A quantitative structure-activity relationship study of N-alkylnorketobemidones and triazinones using structural information content. PMID- 6285932 TI - Homogeneous class of beta-1 adrenergic receptors in rat kidney. Identification by (+/-)-125 iodocyanopindolol binding. AB - The highly specific beta-adrenergic radio-ligand (+/-)-125 iodocyanopindolol (ICYP) was used to characterize the beta-adrenergic receptor subtype present in rat kidney. Binding of ICYP to membranes from rat kidney was of high affinity (KD = 68.9 pM) and saturable with 1.06 pmoles ICYP bound/g tissue wet wt at maximal occupancy of the sites. Analysis of inhibition of ICYP binding by beta 1- and beta 2-selective adrenergic drugs via pseudo-Scatchard ("Hoifstee') plots resulted in linear plots indicating the existence of a homogeneous population of beta-adrenergic receptors. From the resulting KD-values for practolol (2.2 microM), metoprolol (0.21 microM), zinterol (0.4 microM) and IPS 339 (0.046 microM) it is concluded that the beta-adrenergic receptor in rat kidney is of the beta 1-subtype. This subclassification is further supported by the fact that (-) noradrenaline and (-)-adrenaline were equipotent in inhibiting ICYP binding. The beta-adrenergic agonists (-)-isoprenaline and zinterol bind to two distinct states of this beta 1-receptor, a high and low affinity state. GTP (10(-4) M) converts this heterogeneous binding into a homogeneous low affinity binding. PMID- 6285937 TI - [Pharmacokinetics of ceftizoxime with and without probenecid]. AB - Serum concentrations and urinary recovery of ceftizoxime were investigated in 18 healthy volunteers. The substance was given as a bolus (2 min) i.v. injection. In volunteers the injection was preceded by orally administered probenecid, 500 mg, 7 and 1 h before (Group II); the remaining 12 volunteers received medication. 5 min after the injection, the serum concentration in Group I was 107 +/- 22 micrograms/ml and in II 93 +/- 18 micrograms/ml in the mean; 30 min after the injection in I 45 +/- 8 micrograms/ml and 44 +/- 10 micrograms/ml in II. The further of the serum concentration was slower in II than in I: serum concentration 6 h after the injection was 2.6 +/- 0.9 micrograms/ml in I and 4.4 +/- 2 micrograms/ml in II. During the first 8-h period 81% of the administered cephalosporin was recovered in the urine of I and 58% in the urine of II. t/2 beta was calculated for I with 1.76 h and 2.13 h for II. The distribution volume was equal, so that the differences in serum concentrations could be explained by the reduced renal clearance: from 2.1 to 1.4 ml/s. Because the comparison was done with two independent groups and furthermore only 6 volunteers participated in the probenecid group, only the difference in the urine in the first 2 h was statistically significant. PMID- 6285938 TI - Examination of newly synthesized 2-chloroethylnitrosoureas in preterminal leukemia L 5222 and in two transplanted neurogenic tumors. AB - The chemotherapeutic activities of 11 chloroethylnitrosoureas, among them 10 newly synthesized compounds, were investigated in rat leukemia L 5222 and in two neurogenic rat tumors. 1-(4-Amino-2-methyl-5-pyrimidinyl)-methyl-3-(2 chloroethyl)-3-nitrosourea hydrochloride (ACNU, compound 1) and cyclophosphamide (12) served as reference substances. The newly synthesized compounds were 1-(2 chloroethyl)-1-nitroso-3-(2-carboxyethyl)-urea (2-carboxyethyl-CNU, 2), 1-(2 chloroethyl)-1-nitroso-4-methyl-4-formyl-semicarbazide (methyl-formyl-amino-CNU, 3), 1-(2-chloroethyl)-1-nitroso-3-(methylene-2-pyridyl)-urea(methylene-2-pyridyl- CNU, 4), 1-(2-chloroethyl)-1-nitroso-3-(methylene-2-pyridyl)-urea hydrochloride (methylene-2-pyridyl-CNU . HCl, 5) 1-(2-chloroethyl)-1-nitroso-3-(methylene-4 pyridyl)-urea hydrochloride (methylene-4-pyridyl-CNU . HCl, 6), 1-(2-chloroethyl) 1-nitroso-3-(3-pyridino)-urea (pyridyl-3-CNU, 7), 1-(2-chloroethyl)-1-nitroso-3 (3-pyridyl)-urea hydrochloride (pyridyl-3-CNU . HCl, 8), 1-(2-chloroethyl)-1 nitroso-3-[4(2,6-dimethyl-morpholino)]-urea (dimethyl-morpholino-CNU, 9), 1-(2 chloroethyl)-1-nitrosocarbamoyl-morpholine (chloroethyl-nitroso-carbamoyl morpholine, 10), 1-(2-chloroethyl)-1-nitroso-carbamoyl-2,6-dimethyl-morpholine (chloroethyl-nitroso-carbamoyl-dimethylmorpholine, 11). Against both neurogenic tumors cyclophosphamide was distinctly superior to all nitrosoureas. In leukemia L 5222 all nitrosoureas except compounds 7, 8, 11 effected cures. Remarkable differences in toxicity could be observed between the nitroso compounds investigated. PMID- 6285939 TI - [Comparative study on clinical effectiveness of mecillinam and pivmecillinam versus ampicillin in acute urinary infections (author's transl)]. AB - 30 outpatients suffering from acute urinary infections underwent oral treatment with pivaloyloxymethyl-(2S,5R,6R)-6-(perhydroazepin-1 ylmethylenamino)penicillinate (pivmecillinam) (n = 15) or ampicillin (n = 15) in a first study. In a second study 6-beta-(hexahydro-1H-azepin-1-yl)-methylenamino penicillanic acid (mecillinam) (n = 16) or ampicillin (n = 14) were applied i.v. to 30 patients with pretreated urinary infections. The tolerance of the drugs proved to be comparatively good in both studies. The clinical and antibacterial effectiveness were equally good in the first study. However, in the second study the effectiveness of mecillinam However, in the second study the effectiveness of mecillinam seemed to be comparatively inferior. PMID- 6285940 TI - [Familial malignant carotid body chemodectoma with lymph node metastases. Light and electron microscopy study (author's transl)]. AB - A case of malignant carotid body chemodectoma with neighbouring lymph nodes metastases is reported. The case is noteworthy as it was accompanied by another jugulotympanic chemodectoma. There was also hereditary component as four of the patients forebears had been operated for cervical chemodectomas. An ultrastructural study showed the characteristic secretory granules. 32 other cases of tumors of carotid body and jugular glomus with metastases are reviewed. PMID- 6285941 TI - [Rotavirus and prolonged diarrhea syndrome of infants]. PMID- 6285942 TI - Diminished lymphocyte responses to phytohemagglutinin in lung cancer. AB - The PHA response of blood lymphocytes from lung cancer patients was found to be diminished in comparison to normal. Sera from these patients inhibited the blastogenic response of blood lymphocytes from normal subjects. Normal sera could restore to various levels the diminished PHA response of lymphocytes from lung cancer patients. The results suggest that the immunosuppression seen in lung cancer may be mediated by a factor (s) in the serum which might bound reversibly to a certain subpopulation of T-cells and permanently to another and or some other inhibitory mechanism does exist. PMID- 6285943 TI - Fructose 1:6 bisphosphatase activity in the liver and testis of rats and goats. AB - 1. The activity of the enzyme Fructose 1:6 Bisphosphatase (FDPase) was studied in the liver and testis of adult goats and rats. No significant difference in the enzyme activity was observed between liver and testis of rats. Highly significant differences (P less than 0.01) were observed between the activity of goat liver and goat testis, goat liver and rat liver, goat testis and rat testis; the activities being higher in goat tissues. 2. Homogenization of the tissues with water, 0.05 M lactate buffer (pH 3.5), 150 mM KCl and 0.34 M sucrose yielded highest activity with water and lactate buffer followed by Sucrose and KCl. 3. 10 microM of Fe2+ and 45 microM of Zn2+ decreased the enzyme activity of rat testis by 39% and 93% respectively. 4. The rate of hydrolysis of FDP with respect to time in rat liver and testis was a first order reaction. Linear kinetics of the substrate hydrolysis was observed up to 90 min. No substrate inhibition of the enzyme activity was observed up to 50 microM of the substrate. Km of rat liver and testis FDPase were 8.3 microM and 10.5 microM respectively. PMID- 6285944 TI - Sodium nitroprusside and cyanide release. PMID- 6285945 TI - Effects of pindolol and propranolol on beta-adrenergic receptors on human lymphocytes. PMID- 6285946 TI - Biliary excretion of amoxycillin and ceftriaxone after intravenous administration in man. AB - 1 Plasma and biliary concentrations of amoxycillin and ceftriaxone were measured after bolus intravenous administration (500 mg) in four subjects with normal hepato-biliary and renal function. 2 The mean plasma elimination half-life for ceftriaxone (t 1/2 = 330 +/- 30 min) was considerably longer than that for amoxycillin (t 1/2 = 60 +/- 9 min). 3 The biliary concentration of ceftriaxone was above plasma concentration of the drug throughout the study period, whereas amoxycillin concentration in the bile was lower than that in plasma. 4 Both plasma and biliary concentrations of ceftriaxone were substantially higher than previously determined minimum inhibitory concentration (MIC) values for E. coli (and several other common biliary tract pathogens) for over 6 h following drug administration. Amoxycillin concentration in plasma fell below MIC by 2 h, and did not reach inhibitory concentrations in bile. PMID- 6285947 TI - Radiation and nitroimidazoles in supratentorial high grade gliomas: a second clinical trial. AB - As a continuation of a previous controlled trial using "high-dose" metronidazole as a specific sensitizer of hypoxic cells, we used a more efficient nitroimidazole derivative (misonidazole, MISO) in combination with higher doses of radiation in patients with supratentorial high-grade astrocytomas. Sixty-six patients were stratified according to functional level and histological grading, and randomly allocated within 2 weeks of operation of 1 of 3 therapeutic groups: 1, conventional radiation alone; 2, large fractions of radiation with high-dose metronidazole; and 3, radiation as in Group 2 but with equitoxic doses of MISO. We examined survival as the principal end-point of the study. Neither by increasing the dose of radiation over the previous study, nor by using a more efficient sensitizer, were we able to improve survival over the current conventional daily fractionated radiation. PMID- 6285948 TI - Experimental combination and single-agent chemotherapy in human lung-tumour xenografts. AB - A series of human bronchial-carcinoma xenografts (3 small-cell anaplastic, 2 large-cell anaplastic and 3 adenocarcinomas) established in immune-suppressed mice were treated with combination chemotherapy based on clinical regimes. Xenograft response was assessed by the in situ endpoint of growth delay in s.c. tumours. Dose-response relationships of 3 triple-drug combinations and their component agents were explored, allowing the relative contributions of single agents in each combination to be assessed. The results demonstrate that the effects produced in the xenografts were generally consistent with clinical experience. Procarbazine, cyclophosphamide and CCNU stood out as the most effective drugs in small cell carcinoma, but were ineffective in the other histological types. These was some evidence for individuality of therapeutic response among the grafts, supporting the case for incorporating panels of histologically similar xenografts into primary drug-screening programmes to complement existing syngeneic rodent tumour systems. PMID- 6285949 TI - Modulation of adenosine 3', 5'-cyclic monophosphate-dependent protein kinase in skin. AB - Incubation in vitro of full-thickness guinea-pig skin with epinephrine or histamine resulted in the formation of adenosine 3', 5'-cyclic monophosphate (cyclic AMP) and increase in the cyclic AMP-dependent protein kinase activity ratio. The effect was dose-dependent between 10(-5) and 10(-3) M concentration of the agents. Histamine antagonists, diphenhydramine (H1) and cimetidine (H2) prevented stimulation by histamine in a dose-dependent manner suggesting the presence of both types of receptors in full-thickness skin. Cyclic AMP accumulation with a concomitant increase in protein kinase activity denotes that phosphoproteins may have an important role in skin biology. PMID- 6285950 TI - The role of the cellular sodium pump in human granulopoiesis in vitro. AB - The specific sodium pump antagonist ouabain was used to study the effect of sodium pump inhibition on granulopoietic colony formation by normal human bone marrow cells cultured in soft agar for 7 d. Suppression of colony formation was dose-dependent and occurred at a low and reproducible concentration. The use of an extended range of concentrations (1 X 10(-19) M to 1 X 10(-4) M) revealed no additional effect. Inhibition occurred when colony formation was stimulated by either normal human white blood cells in a soft agar underlayer, or the inclusion of pre-formed colony-stimulating activity. Sensitivity to inhibition was similar under either set of conditions, the 50% inhibitory concentrations being 2.70 +/- 0.43 (SE) X 10(-8) M and 2.83 +/- 0.21 (SE) X 10(-8) M respectively. This is interpreted as showing an effect primarily on the colony-forming cells rather than on the cellular production of colony-stimulating activity. Inhibition of colony formation by ouabain was opposed by the addition of extra potassium to the culture medium, confirming that the inhibition was mediated via a perturbation of monovalent cation exchange. Inhibition by 2 X 10(-7) M ouabain appeared to be reversible by simple washing after exposure for up to 24 h, suggesting that the inhibitory effect of ouabain is not primarily cytotoxic. PMID- 6285951 TI - Lithium enhances growth of human leukaemia cells in vitro. AB - Lithium is known to cause leucocytosis in normal humans, and lithium salts have been used therapeutically in attenuating leucopenia in patients undergoing chemotherapy. Recent reports also described leukaemia development during lithium treatment. We have investigated the effect of lithium chloride on the proliferation of human myeloid, erythroblastic, and T- and B-lymphoblast leukaemia cells in vitro. Colony formation by cells of the myeloid leukaemia lines HL-60 and KG-1 was enhanced by lithium chloride, and maximal stimulation was seen at 5 X 10(-4) M. Lithium also increased the proliferation of KG-1a cells, a subline of KG-1 cells that does not respond to colony-stimulating factor, indicating a direct growth-promoting effect on myeloid leukaemia cells. Lithium was found to enhance colony formation by the T-lymphoblast cell line MOLT 4 and the B-lymphoblast line IM-9 at concentrations between 10(-6) and 10(-3) M. The addition of lithium chloride to murine Friend or human K-562 erythroleukaemia cells also caused an augmentation in colony formation. These observations may have relevance to the therapeutic use of lithium in patients with haematological malignancies. PMID- 6285952 TI - Benign monoclonal gammopathy and peripheral neuropathy. AB - Peripheral neuropathy has been described in malignant plasma cell dyscrasias such as multiple myeloma and Waldenstom's macroglobulinaemia. Since it is not known whether the neuropathy is related to the plasma cell disorder or is a paramalignant phenomenon, 21 consecutive out-patients with benign monoclonal gammopathy (BMG) were analysed for peripheral neuropathies. Eleven patients had noticed slight motor and/or sensory extremity symptoms. Clinical examination, electromyographic and electroneurographic studies of the upper and lower extremities were performed. In five patients all results indicated a neuropathy, six other patients had clinical signs of neuropathy and four additional patients had positive electromyographic and/or electroneurographic results compatible with neuropathy. There were no significant differences in haematological parameters between the group where all results indicated a neuropathy and the totally negative group or between the two groups with and without clinical neuropathy. Thus, the benign form of plasma cell dyscrasias seems also to be associated with mild clinical or subclinical peripheral neuropathy. PMID- 6285953 TI - In vitro energy costs of Na+, K+ATPase activity and protein synthesis in muscle from calves differing in age and breed. AB - 1. An in vitro preparation was used to measure rates of oxygen consumption, Na+, K+-ATPase-dependent respiration. [14C]phenylalanine incorporation and tyrosine release of skeletal (sternomandibularis) muscle from 10-21-d-old (three) and 7 month dairy (three) calves and control (CDM; four) and extreme double-muscled (EDM; two) calves. 2. Rates of oxygen consumption was greatest (P less than 0.001) for muscle from 10-21-d-old dairy calves and lowest (P less than 0.05) for CDM calves. 3. Ouabain (10(-6) M) caused a 40% inhibition of muscle respiration. 4. Na+, K+-ATPase-independent respiration was similar for muscle from all calf groups except 10-21-d-old dairy calves which had a value 26% greater (P less than 0.001) than than of older dairy calves. 5. Na+, K+-ATPase-independent respiration was 16% greater (P less than 0.001) for muscle from 10-21-d-old than that of older dairy calves while muscle from EDM calves had a value 11% greater than that of CDM calves. 6. The rate of [14C]phenylalanine incorporation was greater (P less than 0.05) for muscle from 10-21-d-old dairy than from older dairy calves, similar between older dairy and CDM calves, and decreased (P less than 0.05) for EDM calves. 7. Rate of tyrosine release was greatest (P less than 0.05) for muscle from CDM and EDM calves; both dairy groups had similarly low rates of muscle tyrosine release. 8. The energy estimated to be required for peptide bond synthesis accounted for 2.0-3.3% of the O2 consumption of the muscle preparations. PMID- 6285954 TI - THe effects of the gut microflora and dietary fibre on energy utilization by the chick. AB - 1. Groups of adult colostimized chickens were given diets with and without dietary fibre in the form of bagasse or wheat straw. The fibrous materials were analysed for their contents of cellulose, hemicellulose and lignin. The digestible energies (DE) and metabolizable energies (ME) of these diets were measured by a balance method. 2. Groups of germ-free and conventional chicks aged 4 weeks were given diets with and without penicillin or with and without graded levels of wheat straw as a source of fibre and the ME of these diets were determined. 3. The incorporation of the forms of dietary fibre tested reduced the DE and the ME of the diets in adult conventional chickens and the ME in young germ-free and conventional chicks. Both DE and ME were reduced in proportions that suggested that the dietary fibre present was acting largely as an inactive diluent of the dietary energy and was not affecting the absorption of other nutrients. In the young germ-free and conventional chicks, there was a linear relationship between the proportion of wheat straw in one diet and the reduction ME. 4. Dietary penicillin did not alter the ME of the diet in either the germ free or the conventional environments. 5. The reduction of ME with incorporation of wheat straw was less in conventional than in germ-free chicks. It is suggested that this may be due to the ability of chicks with a gut flora to obtain a small amount of energy from wheat straw. PMID- 6285955 TI - A theory of effect of protons and divalent cations on phase equilibria in charged bilayer membranes: comparison with experiment. AB - We summarize the concepts in the recently developed statistical mechanical theory of the effects of proton binding and divalent cation binding on phase equilibria in bilayer membrane composed of acidic phospholipids. The theory is used to calculate membrane phase transition temperatures for different aqueous concentrations of protons, divalent cations and monovalent salt. We discuss methods for calculating transition temperatures even for systems in which there is not an excess of protons or divalent cations relative to lipids. The results are compared with existing experimental data for a number of lipids. There is good agreement between calculated transition temperature vs. pH curves and experimental data for dimyristoylmethylphosphatidic acid, dimyristoylphosphatidylglycerol, dipalmitoylphosphatidylglycerol, dipalmitoylphosphatidylserine, and dimyristoylphosphatidic acid. General thermodynamic considerations are used to derive in Clapeyron-like equation for the rate of variation in membrane transition temperature with divalent cation concentration. This equation and some available experimental data are used to argue that the large increase in solid to fluid phase transition temperature that is observed experimentally as the divalent cation concentration is increased is the result of the metastable solid phase that exists at low but not high divalent cation concentration. A calculated coexistence diagram is compared with existing experimental data for transition temperatures of dimyristoylphosphatidylglycerol membranes at different total calcium concentrations. Good agreement is obtained when the existence of a metastable solid phase is assumed. PMID- 6285957 TI - Hydrodynamic characterization of the Triton X-100 solubilized lactogenic hormone receptor of rat liver. AB - Rat liver plasma membranes, prelabeled with radioactive human growth hormone, were extracted with Triton X-100. The solubilized lactogenic hormone receptor had a Stokes radius of 54.2 A on Sepharose 6B chromatography. Numerical integration of the results of sedimentation experiments on the solubilized hormone-receptor Triton X-100 complex in sucrose/H2O and sucrose/2H2O density gradients produced an s20,w = 5.05 X 10(-13) S and a partial specific volume of 0.791 cm3 g-1. From these data a molecular weight of 148 000 and frictional ratio of 1.40 for the hormone-receptor-Triton X-100 complex were calculated. Triton X-100 was calculated to comprise 32.4% of the complex, and thus, the hormone-receptor complex has a molecular weight of 99 800. These results indicate that the lactogenic hormone receptor, as would be expected of an integral membrane protein, has the capacity to bind a large amount of detergent. PMID- 6285956 TI - Evidence for siroheme-Fe4S4 interaction in spinach ferredoxin-sulfite reductase. AB - Spinach ferredoxin-sulfite reductase (SiR) contains one siroheme and one Fe4S4 center per polypeptide subunit. The heme is entirely in the high-spin Fe3+ state in the oxidized enzyme. When SiR is photochemically reduced with ethylenediaminetetraacetate (EDTA)-deazaflavin, the free enzyme and its CN- and CO complexes show changes in absorption spectra associated with the heme even after the heme has been reduced from the Fe3+ to the Fe2+ state. With CO- or CN- SiR, these spectral changes are associated with the appearance of a classical "g = 1.94" type of EPR spectrum characteristic of reduced Fe4S4 centers. The line shapes and exact g values of the g = 1.94 EPR spectra vary with the nature of the ligand bound to the heme Fe. Photoreduction of free SiR results in production of a novel type of EPR signal, with g = 2.48, 2.34, and 2.08 in the fully reduced enzyme; this signal accounts for 0.6 spin per heme. (A small g = 1.94 type EPR signal, representing 0.2 spin per heme, is also found.) These data suggest the presence of a strong magnetic interaction between the siroheme and Fe4S4 centers in spinach SiR, this interaction giving rise to different EPR signals depending on the spin state of the heme Fe in the reduced enzyme. PMID- 6285958 TI - Stability and components of mature simian virus 40. PMID- 6285959 TI - Laser flash photolysis studies of electron transfer between semiquinone and fully reduced free flavins and the cytochrome c-cytochrome oxidase complex. AB - Laser flash photolysis has been used to determine the rate constants for the reduction of bovine cytochrome oxidase and the cytochrome c-cytochrome oxidase complex by the semiquinone and fully reduced forms of various flavin analogues (FH. and FH-, respectively). Under the condition used, the reaction of FH. with free cytochrome oxidase is too slow to compete with FH. disproportionation whereas FH- reacts measurably. Both FH. and FH- are effective in reducing the complex. The reduction of heme a in the complex is shown to proceed via cytochrome c, and a limiting first-order rate is observed in the case of FH- at high complex concentrations. The data indicate that the interaction site for electron transfer to cytochrome c is the same in the complex as with the free protein, and although a tight complex exists, at least small reactants like the flavins are not sterically hindered in their access to the bound cytochrome c. Moreover, the results also establish that intramolecular electron transfer between cytochrome c and cytochrome oxidase within the complex occurs with a first-order rate constant of greater than 700 s-1. Thus, the presence of cytochrome c greatly enhances electron transfer from reduced flavins to cytochrome oxidase. PMID- 6285960 TI - High-resolution electrophoretic fractionation and partial characterization of the mitochondrial translation products from HeLa cells. PMID- 6285961 TI - Cytochalasin D and platelet gelsolin accelerate actin polymer formation. A model for regulation of the extent of actin polymer formation in vivo. PMID- 6285962 TI - The pathogenesis of Abelson virus lymphomas of the mouse. PMID- 6285963 TI - Avian leukemia viruses. Oncogenes and genome structure. PMID- 6285964 TI - Coulometric and potentiometric evaluation of the redox components of cytochrome c oxidase in situ. AB - A new coulometric-potentiometric titration cuvette is described which permits accurate measurements of oxidation-reduction components in membranous systems. This cuvette has been utilized to measure the properties of cytochrome c oxidase in intact membranes of pigeon breast muscle mitochondria. The reducing equivalents accepted and donated by the portion of the respiratory chain with half-reduction potentials greater than 200 mV are equal to those required for the known components (cytochrome a3 and the high-potential copper plus cytochrome a, 'visible copper', cytochrome c1, cytochrome c, and the Rieske iron-sulfur protein). Titrations in the presence of CO show that formation of the reduced cytochrome a3-CO complex requires two reducing equivalents per cytochrome a3 (coulometric titration). Potentiometric titrations indicate (Lindsay, J.G., Owen, C.S. and Wilson, D.F. (1975) Arch. Biochem. Biophys. 169, 492--505) that both cytochromes a3 and the high-potential copper must be reduced in order to form the CO complex (n = 2.0 with a CO concentration-dependent half-reduction potential, Em). By contrast, titrations in the presence of azide show that the Em value of the high-potential copper is unchanged by the presence of azide and thus azide binds with nearly equal affinity whether the copper is reduced or oxidized. PMID- 6285965 TI - The effect of pH, of ATP and of modification with pyridoxal 5-phosphate on the conformational transition between the Na+-form and the K+-form of the (Na+ +K+) ATPase. AB - An increase in pH decreases the Na+ concentration (Na+ +K+ = 150 mM) necessary for half-maximum activation of the (Na+ +K+)-ATPase at non-saturating concentrations of ATP just as an increase in the concentration of ATP at a given pH. It also decreases the concentration of Na+ necessary for transformation from the K+-form to the Na+-form at equilibrium conditions (Na+ +K+ = 150 mM). An increase in pH increases the rate of the transformation from the K+-form to the Na+-form of the system and decreases the rate of the reverse reaction. The pH effect on the conformation suggests that the K+-form is a protonated form and the Na+-form a deprotonated one. The similarity between the effect of an increase in pH with non-saturating concentrations of ATP and that of an increase in ATP at a given pH suggests that ATP exerts its effect on the transformation from the K+ - to the Na+-form by a decrease in pK values of the system, i.e., by releasing protons, a Bohr effect. Enzyme modified by reaction with pyridoxal 5-phosphate terminated by NaBH4 behaves at a given pH as if it were non-modified enzyme but at a higher pH. The 'pH effect' is seen after modification by pyridoxal 5 phosphate in the presence of ATP, of Na+ without and with ATP, of K+ with ATP but not in the presence of K+ alone. The modification has also a 'pH effect' on the rate of the transformation from the K+ -form to the Na+ -form and on the reverse reaction. There are at least two different pyridoxal 5-phosphate-reactive groups (amino groups), one which can be protected by ATP and which is of importance for activity and another which is not protected by ATP and which is of importance for the pH effect on the conformation. The effect of a protonation-deprotonation of amino groups on the conformation is explained by an involvement of the amino groups in salt bridge formation in between and inside the polypeptide chains, a hemoglobin-like situation. The protonated K+ -form is then a tense T-structure with a high K+, low Na+ affinity and the deprotonated Na+ -form a relaxed, R structure with high Na+, low K+ affinity. ATP facilitates deprotonation by decreasing pK values. Oligomycin has 'pH effect' on the K0.5 for Na+ under equilibrium and steady-state conditions, but oligomycin has no effect on the rate of the transformation from the K+ -form to the Na+ -form, but gives a pronounced decrease of the rate of the reverse reaction, indicating that oligomycin does not react with the K+ -form but with the Na+ -form of the system and prevents the protonation, the E1 to E2 transformation. PMID- 6285966 TI - Identification and partial characterization of plasma membrane polypeptides of Trypanosoma brucei. AB - A plasma membrane-enriched vesicle fraction has been prepared from Trypanosoma brucei by sonication and differential centrifugation on sucrose gradients. This fraction is enriched 5-fold in the plasma membrane marker enzymes adenyl cyclase (EC 4.6.1.1) and ouabain-inhibitable, (Na+ +K+)-dependent adenosine triphosphatase (EC 3.6.1.3). It is also enriched up to 14-fold in iodinated surface proteins, and up to 4-fold in (3H-mannose-labeled glycoproteins, of which the major variable surface coat glycoprotein is the main constituent. Proteins of the plasma membrane fraction and other subcellular fractions have been identified by electrophoretic analysis in sodium dodecyl sulfate-polyacrylamide gradient slab gels. Several high molecular weight surface glycopeptides have been selectively investigated and partially characterized by a combination of metabolic labeling with [3H]mannose, lactoperoxidase-catalyzed surface iodination, and affinity chromatography on Con A-Sepharose. In addition to the major variable surface coat glycoprotein (estimated Mr = 58000), there are several minor surface glycopeptides (Mr = 76000, 86000 and 92000-100000) which are apparent extrinsic membrane components, and two surface glycopeptides (Mr = 42000 and 130000) which are intrinsic membrane components. PMID- 6285967 TI - Evidence for identity between the hexokinase-binding protein and the mitochondrial porin in the outer membrane of rat liver mitochondria. AB - Hexokinase-binding protein and mitochondrial porin were isolated from rat liver mitochondria by different procedures. It was found that the hexokinase-binding protein made lipid vesicles permeable to ADP and formed asymmetric pores in lipid bilayer membranes identical to those obtained from the mitochondrial porin. On the other hand, the mitochondrial porin confers the ability to bind hexokinase. In addition, evidence is presented that both hexokinase-binding protein and mitochondrial porin bind glycerol kinase. PMID- 6285968 TI - Reactivity of glycoconjugates in membranes. I. Determination of transbilayer distribution of gangliosides in lipid vesicles by chemical methods. AB - Two simple chemical methods are described for the determination of the transbilayer distribution of gangliosides GD1a and GM1 in phosphatidylcholine vesicles. The data presented here show an increase in the percentage of GD1a exposed on the outer surface of vesicles with increasing mole fraction of GD1a. The percentage of GD1a exposed on 1:50 and 1:5 GD1a dipalmitoylphosphatidylcholine vesicles were found to be 67% and 83%, respectively. The same trend is seen for vesicles with dimyristoylphosphatidylcholine and distearoylphosphatidylcholine. Extrapolation of the data to infinite dilution gives 65% of GD1a exposed on the surface of GD1a dipalmitoylphosphatidylcholine vesicles. The results indicate that composition dependent changes in transbilayer distribution of GD1a can only partly account for the observed increase in the reactivity of GD1a in vesicles towards neuraminidase from clostridium perfringens as the ratio of GD1a to phosphatidylcholine increases. PMID- 6285969 TI - Regulation of rat brain (Na+ +K+)-ATPase activity by cyclic AMP. AB - The interaction between the (Na+ +K+)-ATPase and the adenylate cyclase enzyme systems was examined. Cyclic AMP, but not 5'-AMP, cyclic GMP or 5'-GMP, could inhibit the (Na+ +K+)-ATPase enzyme present in crude rat brain plasma membranes. On the other hand, the cyclic AMP inhibition could not be observed with purified preparations of (Na+ +K+)-ATPase enzyme. Rat brain synaptosomal membranes were prepared and treated with either NaCl or cyclic AMP plus NaCl as described by Corbin, J., Sugden, P., Lincoln, T. and Keely, S. ((1977) J. Biol. Chem. 252, 3854-3861). This resulted in the dissociation and removal of the catalytic subunit of a membrane-bound cyclic AMP-dependent protein kinase. The decrease in cyclic AMP-dependent protein kinase activity was accompanied by an increase in (Na+ +K+)-ATPase activity. Exposure of synaptosomal membranes containing the cyclic AMP-dependent protein kinase holoenzyme to a specific cyclic AMP-dependent protein kinase inhibitor resulted in an increase in (Na+ +K+)-ATPase enzyme activity. Synaptosomal membranes lacking the catalytic subunit of the cyclic-AMP dependent protein kinase did not show this effect. Reconstitution of the solubilized membrane-bound cyclic AMP-dependent protein kinase, in the presence of a neuronal membrane substrate protein for the activated protein kinase, with a purified preparation of (Na+ +K+)-ATPase, resulted in a decrease in overall (Na+ +K+)-ATPase activity in the presence of cyclic AMP. Reconstitution of the protein kinase alone or the substrate protein alone, with the (Na+ +K+)-ATPase has no effect on (Na+ +K+)-ATPase activity in the absence or presence of cyclic AMP. Preliminary experiments indicate that, when the activated protein kinase and the substrate protein were reconstituted with the (Na+ +K+)-ATPase enzyme, there appeared to be a decrease in the Na+-dependent phosphorylation of the Na+-ATPase enzyme, while the K+-dependent dephosphorylation of the (Na+ +K+)-ATPase was unaffected. PMID- 6285970 TI - ATP/ADP exchange activity of gastric (H+ +K+)-ATPase. AB - The ATP/ADP exchange is shown to be a partial reaction of the (H+ +K+)-ATPase by the absence of measurable nucleoside diphosphokinase activity and the insensitivity of the reaction to P1, P5-di(adenosine-5') pentaphosphate, a myokinase inhibitor. The exchange demonstrates an absolute requirement for Mg2+ and is optimal at an ADP/ATP ratio of 2. The high ATP concentration (K0.5=116 microM) required for maximal exchange is interpreted as evidence for the involvement of a low affinity form of nucleotide site. The ATP/ADP exchange is regarded as evidence for an ADP-sensitive form of the phosphoenzyme. In native enzyme, pre-steady state kinetics show that the formation of the phosphoenzyme is partially sensitive to ADP while modification of the enzyme by pretreatment with 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB) in the absence of Mg2+ results in a steady-state phosphoenzyme population, a component of which is ADP sensitive. The ATP/ADP exchange reaction can be either stimulated or inhibited by the presence of K+ as a function of pH and Mg2+. PMID- 6285971 TI - Absence of 5'-nucleotidase in porcine polymorphonuclear leucocyte membranes. AB - A fraction enriched in plasma membranes from porcine polymorphonuclear leucocytes, isolated by sucrose density centrifugation was shown to possess considerable AMP hydrolysing activity (150 nmol/min per mg protein). However all of this activity could be inhibited using excess p-nitrophenyl phosphate in the incubation medium. Furthermore the hydrolysis of AMP by the membrane was unaffected by the 5'-nucleotidase inhibitor alpha, beta-methyleneadenosine diphosphate and by the lectin concanavalin A, another potent inhibitor of 5' nucleotidase. An antibody against mouse liver 5'-nucleotidase also did not inhibit the activity. These results suggest that the hydrolysis of AMP by porcine polymorph membranes is not accomplished by a specific 5'-nucleotidase and the necessity for distinguishing between true 5'-nucleotidase and non-specific phosphatase activity is discussed. PMID- 6285972 TI - Ion selectivity of the nerve membrane sodium channel incorporated into liposomes. AB - Tetrodotoxin-sensitive sodium channels of lobster nerve membranes were incorporated into soybean liposomes by the freeze-thaw-sonication procedure and their ionic selectivity was studied. Veratridine and grayanotoxin-I were used to activate the sodium channels and the increment of the ionic flux through them was specifically abolished by tetrodotoxin. The drug-sensitive 22Na+, 42K+, 86Rb+ and 137Cs+ influxes were measured. The permeability ratios calculated directly from ion fluxes showed that the channels preferably allow the passage of Na+. No anion influx ([32P]phosphate, [35S]sulfate, 36Cl) sensitive to the drugs was observed. The data reveal that the sodium channels incorporated into liposomes remain cation-selective and discriminate among different cations. PMID- 6285973 TI - Phospholipid transfer between vesicles. Dependence on presence of cytochrome P 450 and phosphatidylcholine-phosphatidylethanolamine ratio. AB - The rate of transfer of spin-labeled phospholipid from donor vesicles of sonicated 1-acyl-2-(10-doxylstearoyl)-sn-glycero-3-phosphocholine to other vesicle was determined as a function of content of cytochrome P-450 and the phosphatidylcholine/phosphatidylethanolamine ratio in the acceptor vesicles. The transfer rate was measured as an increase in intensity that resulted from a decrease in the line width in the EPR spectrum of the spin-labeled phospholipids as they was transferred to the nonspin-labeled acceptor vesicles. A lower transfer rate was observed for acceptor vesicles of pure egg phosphatidylcholine vesicles than for vesicles for a mixture of phosphatidylcholine and phosphatidylethanolamine. The presence of cytochrome P-450 in the acceptor vesicles further increased the transfer rate. Those alterations in the mole ratios of the protein and the two phospholipids that made the bilayer of the reconstituted vesicles more like the membrane of the endoplasmic reticulum resulted in an increase in phospholipid-transfer rate. The mole ratios of components that produce high phospholipid-transfer rates were similar to those that in an earlier study produced a 31P-NMR spectrum characteristic of a nonbilayer phase. These findings suggest that, in the membrane of the endoplasmic reticulum, phospholipid exchange may be an important element in function and interaction with other intracellular organelles. PMID- 6285974 TI - 1 alpha, 25-Dihydroxy-vitamin D-3 regulates ATP-dependent calcium transport in basolateral plasma membranes of rat enterocytes. AB - Basolateral plasma membrane vesicles of rat small intestinal epithelium accumulate calcium through an ATP-dependent pumping system. The activity of this system is highest in duodenum and decreases towards the ileum. This distribution along the intestinal tract is similar as the active calcium absorption capacity of intact intestinal epithelial segments. ATP-dependent calcium uptake in basolateral membrane vesicles from duodenum and ileum increased significantly after repletion of young vitamin D-3-deficient rats with 1 alpha,25-dihydroxy vitamin D-3. Ca2+ -ATPase activity in duodenal basolateral membranes increased to the same extend as ATP-dependent calcium transport, but (Na+ + K+)-ATPase activity remained unaltered. PMID- 6285975 TI - Stimulation of Na+ -dependent amino acid uptake by activation of the Ca2+ dependent K+ channel in the Ehrlich ascites tumor cell. AB - The activation of Ca2+ -dependent K+ channel by propranolol or by ascorbate phenazine methosulphate stimulates Na+ -dependent transport of alpha aminoisobutyric acid. This stimulation arises from a membrane hyperpolarization due to the specific increase of membrane K+ conductance. The same treatment does not modify the Na+ -independent uptake of the norbornane amino acid. PMID- 6285977 TI - A sequence specific endonuclease from Micrococcus radiodurans. AB - A new sequence specific endonuclease, Mra I has been purified from Micrococcus radiodurans. This enzyme cleaves bacteriophage lambda DNA at three sites, adenovirus type 2 DNA at more than 12 sites and has a unique site on phi X174 DNA. It has no sites on SV40, PM2 and pBR322 DNA. The three sites on phage lambda DNA are different from those cleaved by Sma I, Xma I and Xor II. The sites of cleavage are located at 0.424, 0.447 and 0.834 fractional lengths on the physical map of lambda DNA. Mra I is shown to be an isoschizomer of Sac II and Sst II recognizing the palindromic nucleotide sequence '5-CCGC reduced GG-3'. The enzyme shows an absolute requirement of Mg2+, but is active in the absence of added 2 mercaptoethanol. The enzyme shows activity at a broad range of temperature and pH with an optimum at 45 degrees C and pH 7.0. Mra I represents the first restriction enzyme from a bacterium whose DNA lacks modified methylated bases. PMID- 6285976 TI - Molecular cloning and characterization of ribosomal RNA genes from the brine shrimp. AB - A library of genomic DNA from the brine shrimp, Artemia, has been constructed with the Charon 4A phage vector, utilizing EcoRI passenger fragments. Screening this library with purified Xenopus laevis cloned rDNA genes has resulted in the identification and plaque purification of a recombinant containing a complete Artemia (18 S + 26 S) rDNA repeat unit. A physical map derived from the analysis of restriction endonuclease digests of the repeat unit, which measures 13.9 kilobase pairs, is similar to the map derived from genomic DNA. In common with several other species, the 26 S rRNA gene terminates with a HindIII recognition site. PMID- 6285978 TI - Thymidine nucleotide synthesis and catabolism by CHO cells and their changes during the cell cycle. AB - At 0 degrees C, CHO cells efficiently incorporated [3H]thymidine into the nucleotide fraction, but not into DNA. Upon reincubation of asynchronous cultures at 37 degrees C, 15-25% of the radioactivity contained in the cellular nucleotide fraction was released, in the form of thymidine, into the culture medium. At 0 degrees C, however, radioactivity of the nucleotide fraction was retained within the cells. Similarly, dTMP phosphatase (EC 3.1.3.35) in cell extracts was active at 37 degrees C, but not at 0 degrees C, whereas thymidine kinase (EC 2.7.1.21) was active at both temperatures. If synchronous cultures in Gl phase were prelabeled at 0 degrees C and reincubated at 37 degrees C, almost all radioactivity in the nucleotide fraction was released into the medium, whereas in S-phase cultures nearly all radioactivity of the nucleotide fraction was incorporated into DNA. In synchronous S-phase cultures treated with hydroxyurea, radioactivity in the nucleotide fraction was released into the medium at a rate considerably lower than that observed for Gl-phase cells. Rates of endogenous synthesis of thymidine nucleotides were calculated from changes of cellular thymidine nucleotide content, incorporation of thymidine nucleotides into DNA and release of thymidine into the medium during reincubation of prelabeled cultures in thymidine-free medium. The results obtained (see Table III) reveal marked differences between Gl and S phases with respect to the determinants of thymidine nucleotide metabolism. PMID- 6285979 TI - Cloning of genes for bacteriophage T5 stable RNAs. AB - One EcoRI-generated fragment (440 basepairs) and two EcoRI/HindIII fragments (220 and 960 basepairs) from the deletion region of T5 phage have been inserted into the phage lambda XIII and the plasmid pBR322 as vectors. Recombinant DNA molecules were studied by hybridization with in vivo 32P-labeled T5 4-5 S RNAs on nitrocellulose filters. Two-dimensional polyacrylamide gel electrophoretic fractionation and fingerprint analysis of the RNAs eluted from the filters were carried out to identify RNAs coded by cloned fragments. For the accurate localization of the genes for these RNAs, RNA-DNA hybrids were treated with T1 and pancreatic RNAases, and the eluted RNA fragments stable against RNAase action were electrophoresed. It was shown that the EcoRI 440 fragment contains the gene for tRNA 10 (tRNAAsp), the EcoRI/HindIII 220 fragment contains the gene for RNA III (107 bases) and parts of the genes for RNA I (107 bases) and tRNA 12 (tRNAHis), and the EcoRI/HindIII 960 fragment contains only a part of the gene for tRNA 9 (tRNAGln). The arrangement of these genes on the physical map of T5 phage was as follows: -tRNAGln-tRNAHis-RNA III-RNA I-...-tRNAAsp. PMID- 6285980 TI - Restriction enzyme cleavage of ultraviolet-damaged DNA. AB - SV40 and pBR322 DNAs damaged by ultraviolet light were cleaved abnormally by several restriction enzymes because of damage to pyrimidines in the recognition sequences. The use of a tandemly duplicated plasmid provided a particularly sensitive target molecule for detecting pyrimidine dimers and other possible photoproducts. The relative efficiency with which cleavage was blocked (HindIII greater than TaqI greater than EcoRI greater than BamI greater than SalI much greater than Hha I, Hae III) corresponds approximately to the relative frequency of pyrimidine dimer formation in the recognition sequences, but at a slightly higher frequency in potential sites for the non-cyclobutane T-C product. The pyrimidine dimers appear to have a range of influence that extends 1 to 3 basepairs along the DNA molecule. These effects provide clues to the way DNA damage from mutagens and carcinogens can interfere with specific enzyme-DNA interactions. PMID- 6285981 TI - Solution and ion-complexed conformations of beauvericin determined by proton magnetic resonance spectroscopy. AB - The conformational properties of the cyclohexadepsipeptide antibiotic Beauvericin have been investigated by 1H-NMR spectroscopy in polar (C2H3O2H) and non-polar (CCl4, C62H6, C2HCl3) solvents and in two solvent mixtures; one a mixture of a polar and non-polar solvent (C2H3O2H/CCl4) and the other an aromatic solvent in a non-polar environment (C62H6/CCl4). The ion-complexation properties of Beauvericin with alkali metal halides (Li+, Na+, K+, Cs+) have also been studied. It is demonstrated that changes in chemical shifts of Beauvericin with concentration, with polarity of solvent or with added alkali metal ion reflect changes not only in the solvent properties but also changes in backbone conformation and changes due to ion-complexation, where appropriate, and therefore cannot be used, by themselves, to determine the conformation of the molecule, its self-aggregation properties, or the stoichiometry of the metal ion complex. The backbone conformations of Beauvericin in different environments are determined by methods that are independent of chemical shift analysis; i.e., by measurements of 5J(HH) magnitudes observed between the alpha-CH protons of the L phenylalanine and D-hydroxyisovaleric acid (DHyIv) residues and by nuclear Overhauser effect measurements observed between alpha-CH(HyIv) and (N)-CH3(Phe) proton signals. In the knowledge of these results the chemical shifts of Beauvericin in different environments can then be rationalised. It is found that the conformation of Beauvericin in a polar solvent is different from that found in a non-polar solvent and from that found for the in the ion-complexed form is similar to that found in non-polar solvents. By taking into account the conformational properties of the L-phenylalanine and DHyIv side-chains, it is possible to assign unambiguously the magnetically non-equivalent beta-CH2(Phe) and gamma Me(HyIv) proton signals and so elucidate the complete conformational behaviour of the uncomplexed forms of Beauvericin in a polar and a non-polar environment, and of the ion-complexed form of Beauvericin in a polar solvent. PMID- 6285982 TI - Studies on Scapharca hemoglobins. Properties of the dimeric protein reconstituted with Fe- or Co-porphyrin. AB - A native globin from the dimeric hemoglobin, hemoglobin I, of the mollusc Scapharca inaequivalvis has been obtained with the acid-acetone method. The globin has a lower sedimentation coefficient than the native protein at neutral pH; its reconstitution product with natural heme has the same physicochemical and functional properties as the native protein. proto- and meso-cobalt hemoglobin I have been prepared and characterized. proto-Cobalt hemoglobin I binds oxygen reversibly with a lower affinity and a lower cooperativity than native hemoglobin I; thus, the changes in the functional properties brought about by substitution of iron with cobalt are similar to those observed in human hemoglobin A. The EPR spectra of deoxy-proto-cobalt hemoglobin I and of the photolysis product of oxy meso-cobalt hemoglobin I indicate that two histidine residues are the apical heme ligands. The broad signal at g = 2.38 in deoxy-proto-cobalt hemoglobin I points to a constrained structure of the heme site in this derivative which results from a distorted coordination of the hindered proximal histidine. A similar structure has been proposed previously for the alpha chains in deoxy-cobalt hemoglobin A. PMID- 6285983 TI - The influence of 2-oxoglutarate on the activity of prolyl 4-hydroxylase. AB - 1. Treatment of prolyl 4-hydroxylase (prolyl-glycyl-peptide, 2 oxoglutarate:oxygen oxidoreductase (4-hydroxylating), EC 1.14.11.2) with 2 oxoglutarate in the absence of added Fe2+ for 10 s causes partial inactivation of the enzyme which is not reversed by subsequent addition of Fe2+. It appears that 2-oxoglutarate prevents loss of enzyme-bound iron and prevents access of added iron to its binding site. 2. For optimal enzyme activity the enzyme should be preincubated for 15 s with Fe2+ (5 microM). 3. Under turnover conditions prolyl 4 hydroxylase does not release iron. 4. The inactivation brought about by pre incubation with 2-oxoglutarate and O2 in the absence of ascorbate is partly reversed by removal of 2-oxoglutarate. 5. It is proposed that dead-end complex formation with 2-oxoglutarate is responsible for the inactivation of the enzyme by 2-oxoglutarate in the absence of either ascorbate or Fe2+. 6. Optimal enzyme activity is obtained if the reactants are added to the reaction medium in the following order: enzyme, iron and ascorbate in any order, then after at least 15 s 2-oxoglutarate and finally (Pro-Pro-Gly)5 to start the reaction. PMID- 6285984 TI - Prolyl 4-hydroxylase activity in relation to the oxidation state of enzyme-bound iron. The role of ascorbate in peptidyl proline hydroxylation. AB - In agreement with others (Myllyla, R., Kuutti-Savolainen, E.-R. and Kivirikko, K.I. (1978) Biochem. Biophys. Res. Commun. 83, 441-448), it was found that, in the absence of ascorbate, prolyl 4-hydroxylase (prolyl-glycyl-peptide, 2 oxoglutarate:oxygen oxidoreductase (4-hydroxylating), EC 1.14.11.2) catalyses the hydroxylation of peptidyl proline, stoicheiometrically coupled to the oxidative decarboxylation of 2-oxoglutarate, at a high initial rate. Under these conditions the enzyme becomes inactivated by at least 90% within 1 min in the presence of 400 microM 2-oxoglutarate, in the presence or absence of the peptide substrate (Pro-Pro-Gly)10. The enzyme can be partly reactivated by ascorbate, but not by Fe2+. Addition of a stoicheiometric amount of iron to the enzyme gives rise to a small EPR signal at g = 4.3, which is typical of a high-spin d 5 ion in a rhombic environment. After subsequent incubation for 30 s at 37 degrees C in the presence of 2-oxoglutarate, the amplitude of the EPR signal at g = 4.3 increases 3-4-fold and corresponds to virtually all of the iron added. In addition, an EPR signal at g = 2.0 is formed under these conditions. The signal at g = 4.3 decreases after subsequent addition of ascorbate. It is concluded that in the presence of 2 oxoglutarate enzyme-bound Fe2+ is rapidly converted to Fe3+, leading to inactivation of the enzyme. Enzyme-bound Fe3+ can be reduced again by ascorbate, thus reactivating the enzyme, or, in the absence of 2-oxoglutarate, by Fe2+. PMID- 6285985 TI - Acetyl glycerylphosphorylcholine inhibition of prostaglandin I2-stimulated adenosine 3',5'-cyclic monophosphate levels in human platelets. Evidence for thromboxane A2 dependence. AB - Previous studies with AGEPC (1-O-hexadecyl/octadecyl-2-acetyl-sn-glyceryl-3 phosphorylcholine) stress the independence of the proaggregatory activity of AGEPC from the platelet cyclooxygenase. However, our dose response analyses in human platelet-rich plasma show distinct primary and secondary waves of aggregation in response to AGEPC. Second wave aggregation is inhibited completely by either 10 micro M indomethacin, a cyclooxygenase inhibitor, or 5.6 micro M 9,11-azoprosta-5,13-dienoic acid, a thromboxane A2 synthetase inhibitor. Simultaneous addition of AGEPC and prostaglandin I2 to platelet-rich plasma results in a marked increase in platelet cyclic AMP, which is not different from the prostaglandin I2 response alone. However, if prostaglandin I2 is added to AGEPC-stimulated platelets at a point where secondary aggregation is just beginning, AGEPC can attenuate prostaglandin I2-stimulated cyclic AMP accumulation. The inhibition by AGEPC is blocked by either cyclooxygenase or thromboxane A2 synthetase inhibitors, and radioimmunoassay of thromboxane B2 confirmed that the inhibition of prostaglandin I2-stimulated cyclic AMP accumulation is due to thromboxane A2 synthesis, and that AGEPC-stimulated secondary aggregation does not start until thromboxane A2 is synthesized. These data suggest that much of the bioactivity of AGEPC is attributable to thromboxane A2. PMID- 6285986 TI - Effect of salts on membrane binding and activity of adipocyte phosphatidate phosphohydrolase. AB - Isosmotic replacement of sucrose in a low ionic strength homogenizing buffer (0.25 M sucrose/1 mM EDTA/1 mM Tris-HCl, pH 7.4) with KCl increased the microsomal and decreased the soluble phosphatidate phosphohydrolase (EC 3.1.3.4) activity of isolated rat fat cells. At 54 mM KCl the microsomal specific activity was increased 6-fold and the soluble activity was decreased to less than one third. Binding of enzyme was promoted by KCl and NaCl when the once isolated soluble and microsomal fractions were recombined and incubated at 37 degrees C. Half-maximal binding occurred at about 17 mM salt and maximal binding at about 50 mM. The pH optimum of binding was 7.8 in 15 mM Hepes. MgCl2, CaCl2 and spermine prevented desorption of microsomal enzyme at mu molar levels and maximal effects were observed at concentrations below the 1 mM level. At maximum, however, the prevention of desorption was less by these salts than it was by KCl. MgCl2 and spermine also interfered with the effect of KCl. Moderate salt-induced loading of microsomes with the phosphohydrolase (specific activity increased 3.5-fold) increased their ability to incorporate 14C into triacylglycerol from sn-[U 14C]glycerol 3-phosphate while a high loading (specific activity increased 6 fold) had no effect or even suppressed it. The results are discussed in relation to a role of translocation of phosphatidate phosphohydrolase in glyceride biosynthesis and its control. PMID- 6285987 TI - Evidence for a relative excess of lysyl hydroxylase in chick embryo tendon and cartilage compared with bone and skin. AB - Various chick embryo tissues were incubated in vitro with a range of Zn2+ concentrations, and inhibition of the hydroxylations of collagen proline and lysine residues was studied in the intact tissues. At an constant inhibition level of proline hydroxylation, lysine hydroxylation proved to be inhibited more in the skin and bone than in the tendon and cartilage. The ratios of lysyl hydroxylase to prolyl hydroxylase activity were also distinctly lower in the former than in the latter. The variations observed in the reduction of lysine hydroxylation by Zn2+ thus correlate well with the differences seen in the enzyme activity ratios. No differences are found in the Ki values for Zn2+ between purified prolyl and lysyl hydroxylases or between the same enzymes from different tissues, and consequently the differences in lysine hydroxylation inhibition between the various tissues cannot be explained by differences in the kinetic constants. Recent studies also suggest that the existence of tissue-specific lysyl hydroxylase isoenzymes is improbable. The data thus suggest that there is a relative excess of lysyl hydroxylase activity in tissues such as tendon and cartilage, in which the lysine hydroxylation was less sensitive to Zn2+ inhibition, compared with skin and bone, where lysine hydroxylation was inhibited to a greater extent. These data are in a good agreement with the findings concerning variation in the reduction in lysine hydroxylation in different tissues with age or in the Ehlers-Danlos Syndrome Type VI. PMID- 6285988 TI - Effects of carbohydrate removal on the structure and activity of bovine lutropin. AB - Bovine lutropin was successfully deglycosylated by treatment with anhydrous HF at 0 degrees C for 60 min. The overall loss of carbohydrate residues was about 70%. Fucose, mannose and galactosamine residues were removed completely. About six residues of N-acetylglucosamine were left in the deglycosylated hormone. This degree of deglycosylation did not reduce receptor binding ability but in vitro biological activity was completely lost. Consistent with these properties, the deglycosylated lutropin inhibited the in vitro biological action of the native lutropin. The results demonstrate that the full complement of carbohydrate residues including the sulfated hexosamines are not required for receptor binding but they are necessary to impart appropriate conformational features necessary for activation of the target cells. PMID- 6285989 TI - Lack of stimulation of renal (Na+ +K+)-ATPase by thyroid hormones in the rabbit. AB - The effect of 1-3,5,3'-triiodothyronine (T3) and thyroxine (T4) on (Na+ +K+) ATPase activities was examined in rabbit kidneys because in this tissue almost 80% of the metabolism is connected to active sodium transport. T3-receptor concentrations were estimated as 0.62 and 0.80 pmol/mg per DNA in the cortex and outer medulla, respectively. A dose of 0.5 mg T3/kg body weight for 3 days increased basal metabolic rate by almost 60%, and the mitochondrial 1-alpha glycerophosphate dehydrogenase activity was increased by 50% in both the cortex and medulla. (Na+ +K+)-ATPase activity in the liver was raised by almost 50%. However, no changes in (Na+ +K+)-ATPase activities or binding sites for [3H]ouabain in either the kidney cortex or medulla could be observed. T4 at 16 mg/kg daily for 14 days was also without effect on renal (Na+ +K+)-ATPase activities. Furthermore, the response to T3 was absent at high sodium excretion rates induced by unilateral nephrectomy and extracellular volume expansion. Thus, despite stimulation of basal metabolic rate and renal 1-alpha-glycerophosphate dehydrogenase activity by T3 and T4, the (Na+ +K+)-ATPase activity in the rabbit kidney is identical in euthyroid and hyperthyroid states. However, thyroid hormones prevent the normal natriuretic response to extracellular volume expansion. PMID- 6285990 TI - Further purification and some properties of a gelatin-specific proteinase of human leucocytes. AB - A latent gelatin-specific proteinase (gelatinase) was isolated from the crude extract of human leukocytes. The enzyme was purified about 180-fold (7040 units/mg) with an overall yield of 23%. The isolated protein migrated as a single band on sodium dodecyl sulfate polyacrylamide-gel electrophoresis. Its mobility was unaffected by reducing agent. The protein band corresponded to approx. 90-94 kDa. Gelatinase activity was strongly inhibited by chelating agents, such as EDTA and 1,10-phenanthroline. This inhibition was reversed by Zn2+ and Co2+; other metal ions were less or not at all effective in reversing the inhibition. Moreover, Co2+ stimulated gelatinase activity. These results indicate that Zn2+ and/or Co2+ are essential for the activity of this gelatinase. PMID- 6285991 TI - Evidence for a defect in the number of beta-adrenergic receptors and in the adenylate cyclase responsiveness to guanine nucleotides in fat cells after adrenalectomy. PMID- 6285992 TI - Effect of external ATP on the plasma membrane permeability and (Na+ +K+)-ATPase activity of mouse neuroblastoma cells. AB - 1. Addition of 3.5 mM ATP to mouse neuroblastoma Neuro-2A cells results in a selective enhancement of the plasma membrane permeability for Na+ relative to K+, as measured by cation flux measurements and electro-physiological techniques. 2. Addition of 3.5 mM ATP to Neuro-2A cells results in a 70% stimulation of the rate of active K+ -uptake by these cells, partly because of the enhanced plasma membrane permeability for Na+. Under these conditions the pumping activity of the Neuro-2A (Na+ +K+)-ATPase is optimally stimulated with respect to its various substrate ions. 3. External ATP significantly enhances the affinity of the Neuro 2A (Na+ +K+)-ATPase for ouabain, as measured by direct [3H]ouabain-binding studies and by inhibition studies of active K+ uptake. In the presence of 3.5 mM ATP and the absence of external K+ both techniques indicate an apparent dissociation constant for ouabain of 2 X 10(-6)M. Neuro-2A cells contain (3.5 +/- 0.7) X 10(5) ouabain-binding sites per cell, giving rise to an optimal pumping activity of (1.7 +/- 0.4) X 10(-20) mol K+/min per copy of (Na+ +K+)-ATPase at room temperature. PMID- 6285993 TI - Alamethicin or detergent permeabilization of the cell membrane as a tool for adenylate cyclase determination. AB - Treatment of mouse lymphocytes with very low concentrations of alamethicin or Lubrol PX induces spontaneous permeabilization of the plasma membrane to ATP and allows determination of adenylate cyclase activity in whole cells. The permeabilized cells retain responsiveness to hormones (isoproterenol, adenosine analogs) and to fluoride. The main advantage of this new method is that it does not require any homogenization step, and thus adenylate cyclase activities can be accurately and reproducibly measured with very low amounts of cells. It should be especially useful for the study of purified lymphocyte subpopulations. PMID- 6285994 TI - The periodic change in adenosine 3',5'-monophosphate concentration in sea-urchin eggs. AB - The level of adenosine 3',5'-monophosphate (cyclic AMP) in the eggs of the sea urchin, Anthocidaris crassispina, was found to change periodically after fertilization. The minimum and maximum levels of cyclic AMP were 1.0 X 10(-7)M and 1.5 X 10(-6)M, respectively. The activity of adenylate cyclase in a 105 000 X g precipitate reached a plateau at 20 min after fertilization and stayed constant for at least 2 h. It was also found that 1.0 mM CaCl2 increased the activity of adenylate cyclase in the same precipitate from unfertilized eggs. In contrast, phosphodiesterase activity changed periodically and correlated with cyclic AMP levels in the eggs. Up to a concentration of 1.5 X 10(-6)M cyclic AMP, phosphodiesterase activity was low, but it became activated when the level of cyclic AMP rose beyond this level. These results indicate that the change in the intracellular level of cyclic AMP is regulated mainly by the change in phosphodiesterase activity. PMID- 6285995 TI - Stimulation of the insulin secretory mechanism following barium accumulation in pancreatic beta-cells. AB - Electrothermal atomic absorption spectroscopy was employed for measuring barium in beta-cell-rich pancreatic islets microdissected from ob/ob-mice. Both the uptake and efflux of barium displayed two distinct phases. There was a 4-fold accumulation of barium into intracellular stores when its extracellular concentration was 0.26 mM. Unlike divalent cations with more extensive intracellular accumulation, the washout of Ba2+ was not inhibited by D-glucose. Ba2+ served as a substitute for Ca2+ both in maintaining the glucose metabolism after removal of extracellular Ca2+ and making it possible for glucose to stimulate insulin release. Furthermore, Ba2+ elicited insulin release in the absence of glucose and other secretagogues. The latter effect was reversible and was markedly potentiated under conditions known to increase the beta-cell content of cyclic AMP. It is likely that the observed actions of Ba2+ are mediated by Ca2+, since Ca2+ -dependent regulatory proteins, such as calmodulin, apparently cannot bind Ba2+ specifically. PMID- 6285996 TI - Clinical response to clonidine and imipramine, and platelet alpha 2 adrenoreceptors in a case of agoraphobia with panic attacks. PMID- 6285997 TI - Tricyclic antidepressants induce sphingomyelinase deficiency in fibroblast and neuroblastoma cell cultures. AB - Tricyclic antidepressants (imipramine and desipramine) gave rise to an important decrease of sphingomyelinase activity in murine neuroblastoma and human fibroblast cell cultures. It occurred within 1 to 2 hours at a final concentration of 1 or 2 X 10(-5) M in cell culture medium. Other lysosomal enzymes such as acid lipase, arylsulfatases A and B and hexosaminidases were not modified. Low level of sphingomyelinase activity may be related to the amphiphilic characteristics of the drugs: iminodibenzyle which has the same tricyclic core but is devoid of the side chain necessary for amphiphilic properties had no effect. As iminodibenzyle has no therapeutic action, amphiphilic may be requisite to antidepressant properties of tricyclic drugs. PMID- 6285998 TI - Coupling of water and ion fluxes in a K+-selective channel of sarcoplasmic reticulum. AB - Streaming potentials arising across a K+-selective channel from fragmented sarcoplasmic reticulum were measured by incorporating the channel into planar bilayer membranes and imposing osmotic gradients across the membranes by addition of sorbitol or urea to only one side. Single-channel zero-current potentials were determined, and dilution artifacts were corrected for by addition of valinomycin to the bilayer. The streaming potentials were found to be unusually small, 1.1 mV per osmolal. The potentials were linearly related to the osmotic gradient across the bilayer, and were identical for sorbitol and urea. The results imply that the channel cannot be envisioned as a long tube, like gramicidin, but rather as a short constriction of less than 10 A in length opening out into wider mouths on either side of the membrane. PMID- 6285999 TI - Protein conformation from electron spin relaxation data. AB - Electron spin relaxation data from five ferric proteins are analyzed in terms of the fractal model of protein structures. Details of this model are presented. The results lead to a characterization of protein structures by a single parameter, the fractal dimension, d. This structural parameter is shown to determine the temperature dependence of the Raman electron spin relaxation rate, which varies as T3 + 2d. Computations of d are made using x-ray data for 17 proteins. The results range from d = 1.76 for lysozyme to d = 1.34 for ferredoxin. These values are compared with values of d obtained from the present electron spin relaxation data on five ferric proteins. Typical results are d = 1.34 +/- 0.06 from relaxation data and 1.34 +/- 0.05 from x-ray data for ferredoxin; d = 1.67 +/- 0.03 from relaxation data and 1.66 +/- 0.05 from x-ray data for ferricytochrome c. The data thus support the theoretical model. Applications of this spin resonance technique to the study of changes in protein conformation are discussed. PMID- 6286000 TI - Stability constant of the 1:1 complex of sodium with guanosine 5'-monophosphate. AB - The stability of the 1:1 complex of sodium ion with the dianion of guanosine 5' monophosphate has been determined by means of a potentiometric titration employing a specific ion electrode. The stability constant for the reaction Na(+) + 5'-GMP(2-) Na(5'-GMP)(-) was found to be 2.85 +/- 0.36 M(-1) at 5 degrees C and an ionic strength of 1.1 +/- 0.1 M. Although 5'-GMP forms ordered self-structures at high concentration in the presence of sodium ions, in dilute solution and at low sodium ion concentrations the Na(+) binding is weak and typical of that for other nucleotides. PMID- 6286001 TI - Selectivity of the Ca2+-activated and light-dependent K+ channels for monovalent cations. AB - The ionic selectivity of the Ca(2+)-activated K(+) channel of Aplysia neurons and of the light-dependent K(+) channel of Pecten photoreceptors to metal and organic cations was studied. The selectivity sequence determined from reversal potential measurements is T1(+) K(+) > Rb(+) > NH(+) (4) > Cs(+) > Na(+), Li(+) and is identical to the sequence determined previously for voltage-dependent K(+) channels in a variety of tissues. Our results suggest that some physical aspect of the K(+) channel is conserved in phyllogenetically different tissues and cells. PMID- 6286003 TI - Radical scavenging and electron-transfer reactions in Polyporus versicolor laccase a pulse radiolysis study. AB - The interaction of the radicals OH, t-BuO, Eaq, CO2 and O2 with the copper oxidase, laccase, from Polyporus, has been studied by the pulse-radiolysis technique. Each of these radicals formed transient adducts with a broad absorption maximum around 310 nm. Analysis of the optical properties and of the very fast rates of formation of these compounds shows that each radical interacts with a limited number of sites on the polypeptide part of the protein amongst R-S S-R, histidine and aromatic residues. Interaction with the carbonyl group of some of the peptides bonds is also possible. The few target sites are probably hit simultaneously and electron transfer between these sites may also occur. In all cases, ina subsequent step, intramolecular electron transfer from the polypeptide radical adducts leads to a partial reduction of the blue type-1 Cu2+ with rates varying between 10(3) adn 10(4) s-1. Further reduction of the type-1 CU2+ occurs through a slow intermolecular reaction between two laccase radical transient adducts. In the case of CO2 and O2, this slow reduction could alternatively be due to an intermolecular reaction between laccase and CO2 or O2. The oxidation radicals OH, Br2 and (SCN)2, which formed radical adducts with fully ascorbate reduced laccase, did not induce any type-1 copper reoxidation. PMID- 6286002 TI - Dependence of ion flow through channels on the density of fixed charges at the channel opening. Voltage control of inverse titration curves. AB - Model calculations were done to investigate the effect of titratable fixed charges at channel openings on ion flow through open channels. The current titration curves (channel current vs. bulk pH) can assume the shape expected from the change of the ionic surface concentration with pH (c-control), or be inverted, i.e., follow the change of the electrical field within the membrane (V control). The relationships were explored pars pro toto for Goldman-Hodgkin-Katz channels, two-barrier one-site channels and six-barrier five-site channels. With net current flowing in the direction of the concentration gradient and from the titrated fixed charge layer into the channel, c-control is the sign of low channel occupancy (entrance-step limitation) and V-control the sign of high channel occupancy (exit-step limitation). At intermediate occupancy, the current titration curve can be nearly invariant to pH. PMID- 6286004 TI - Fluorescence quenching and spin label electron spin resonance studies of stacking self-association in aqueous solutions of 2-aminopurine riboside and its 5'-mono- and -diphosphate. AB - The autoassociation of 2-aminopurine riboside (rn2Pur) and its 5'-mono- (P rn2Pur) and 5'-diphosphate (PP-rn2Pur) in neutral aqueous solutions was investigated using fluorescence quenching and ESR spin-label methods within the range 276-358 K. Respective equilibrium constants and thermodynamic functions were derived therefrom assuming two models of infinite autoassociation: (i) an isodesmic one (K2 = K3 = ... Kp), and (ii) one in which K2 no equal to K2 = K4 ... Kp. Comparative analysis of these data and that of the parent 2-aminopurine, obtained previously, allowed us to formulate the following conclusions: (1) the mechanism of autoassociation of rn2Pur varies with temperature in such a way that a T = 318 K the isodesmic model is fulfilled (K2 = Kp); at high temperatures Kp/K2 greater than 1, i.e. the process is cooperative, while at lower temperatures it becomes anticooperative (Kp/K2 greater less than 1); (2) at 298 K the tendency to autoassociation decreases in the order; rn2Pur greater than P rn2Pur greater than PP-rn2Pur; (3) rn2Pur forms highly packed complexes with the bases stacked and the ribofuranose residues interacting via hydrogen bonds or water bridges; (4) autoassociation of P-rn2Pur and PP-rn2Pur is mainly governed by stacking of the bases, while the ribose phosphate residues attain a trans configuration corresponding to the lowest electrostatic repulsion between charged phosphate groups; even at high ionic strength (I = 0.8), a positive electrostatic contribution to the free enthalpy of autoassociation is observed; (5) the two methods employed gave similar results for P-rn2Pur, but somewhat different ones for rn2Pur because the presence of the spin label (nitroxide stable radical) at the 2'(3')-OH group of the ribose residues prevents its interaction via hydrogen bonding with an unlabeled one of an adjacent nucleoside. PMID- 6286005 TI - Measurement of intracellular pH. PMID- 6286007 TI - Simultaneous presence of terminal adenylyl, cytidylyl, guanylyl, and uridylyl transferase in healthy tomato leaf tissue: separation from RNA-dependent RNA polymerase and characterization of the terminal transferases. AB - The presence of terminal nucleotidyl transferase activities catalyzing the addition of AMP, CMP, GMP, and UMP residues to the 3' ends of oligonucleotide primers was detected in healthy tomato plants. These enzyme activities copurify with RNA-dependent RNA polymerase during the initial stages of purification. Their separation from RNA-dependent RNA polymerase is finally achieved by DEAE chromatography: terminal transferase activities are retained on DEAE while RNA dependent RNA polymerase does not bind in the presence of 20 mM MgCl2. Elution by a linear gradient of 0 to 400 mM NH4Cl releases all four terminal transferase activities from the DEAE column at a concentration of 270 mM NH4Cl, thus suggesting that they may belong to one enzyme molecule; this question, however, needs further clarification. The enzyme activities are completely dependent on the presence of an RNA primer and are strongly influenced by its base composition as well as its chain length. Characterization of the respective reaction products by electrophoresis on 15% polyacrylamide sequencing gels reveals striking differences as to the number of nucleotides added to a given primer. In the case of UMP transfer to U8 or A8 and in the case of GMP transfer to A8 only 1 to 6 nucleoside monophosphates are added to the 3' terminus of the oligonucleotide primer, whereas in the case of AMP transfer to A8 or U8, the CMP transfer to A8, and the GMP transfer to U8, longer chains of minimally 30 nucleotides are added to the respective primer. After gradient elution from DEAE the transferase preparation displays no nucleolytic activity when incubated in the presence of 3H labelled ribosomal RNA or [3H]poly(A) X poly(U). Only in the case of [3H]poly(A) and [3H]poly(U) or [3H]poly(C) 10 to 15% of the radioactivity is transferred to acid-soluble counts. PMID- 6286006 TI - Effect of manganese ions on the interaction between ribosomes and endoplasmic reticulum membranes isolated from rat liver. PMID- 6286008 TI - Microinjection of cyclic nucleotides provides evidence for a diffusional mechanism of intraneuronal control. AB - Cyclic 3',5'-AMP and cyclic 3',5'-GMP injected into large neurons of the snail Helix lucorum altered neuron activity. The effect of cAMP is usually depolarizing and that of cGMP hyperpolarizing. The results are specific for 3',5'-cyclic nucleotides. The experiments support the hypothesis that reaction-diffusion processes involving cyclic nucleotides form the basis of an intraneuronal system of information processing. PMID- 6286010 TI - Retroviruses as etiologic agents of some animal and human leukemias and lymphomas and as tools for elucidating the molecular mechanism of leukemogenesis. PMID- 6286009 TI - On fundamental and phenomenological models, structure and mechanical properties of collagen, elastin and glycosaminoglycan complexes. AB - This paper reviews the structure (from molecular to macroscopic level) of collagen, elastin and glycosaminoglycans, with special reference to their functional properties and to how their behavior can be elucidated from in vitro manipulations of the tissues by biochemical means. Modes to analyse the mechanical behavior are briefly discussed and exemplified by data from reconstituted collagenous "tissue". It is concluded that the basic equation for the "non-linear elasticity" component in Fung's law (and for the reconstituted "tissue" a modification of it) is a powerful tool for analysis of the physiological range of the stress-strain curve. Further, enzymatic "dissection" of one tissue element at a time provides a valuable method for the analysis of tissue element interactions. This approach is illustrated with data from aortic tissue. It is shown that the mechanical properties of the aorta depend on the interaction between elasticity and collagenous elements and that the strength of the tissue is not derived from its collagen fibers per se. PMID- 6286011 TI - Analysis of T lymphocytes after bone marrow transplantation using monoclonal antibodies. AB - Using monoclonal antibodies to cell surface antigens and fluorescent cell sorter analysis, we studied peripheral blood lymphocyte subsets after bone marrow transplantation (BMT). In 13 patients studied 3 mo or more after BMT, the ratio of T-cell subsets defined by antibodies OKT4 and OKT8 was reversed (OKT4/OK%8 = 0.7 +/- 0.3) in comparison to normal volunteers or bone marrow donors (ratio OKT4/OKT8 = 1.7 +/- 0.4) (p less than 0.001). This reversed ratio persisted for up to 3 yr after BMT. In contrast to a previous report, presence of an abnormal ratio of T-cell subsets did not correlate with clinically significant graft versus-host disease (GVHD). In agreement with a previous study, (26% +/- 8%; less than 4% in normals (p less than 0.001) and antibody OKT10 reactive cells (39% +/- 20% versus 10% +/- 4%) (p less than 0.01), suggesting in vivo activation. However, their PBL did not react with antibody B3/25 (antitransferrin receptor), a marker found on normal PBL after in vitro activation by mitogens (BMT patients less than 5%; normal PBL T cells plus PHA 45% +/- 11%). These results demonstrate that BMT patients have: (A) an abnormal ratio of T-cell subsets in the presence or absence of clinically significant GVDH disease so that these measurements were not useful in monitoring patients; (B) an increased number of T cells with cell surface phenotype (OKT8+, Ia+, OKT10+, B3/25-) that is distinct from normals but similar to patients with infectious mononucleosis or acquired hypogammaglobulinemia. PMID- 6286012 TI - Cellular and extracellular myeloperoxidase in pyogenic inflammation. AB - We explored the effect of in vitro phagocytosis and in vivo inflammation on the MPO content of functioning neutrophils and on the ability of these cells to export active MPO into the extracellular environment. After ingestion of staphylococci, neutrophils retained 52% of their MPO and released 8% into the medium in active form; the remaining 40% of their MPO could no longer be detected. During bacterial infection induced by intradermally injecting staphylococci, neutrophils harvested from minced infected lesions contained 52% of the MPO of circulating neutrophils that had not reached the lesions. Extracellular fluid from the lesions contained active MPO secreted by the neutrophils, and concentrations of 10-45 U/ml were detected. These data demonstrate that functioning neutrophils can lose approximately half of their MPO. In vitro, 4%-8% of neutrophilic MPO appears in the extracellular space and 40% is inactivated. In vivo, the MPO content of inflammatory neutrophils also decreases, and MPO appears in the extracellular fluid in active form where it is available to participate in a variety of physiologic processes. PMID- 6286013 TI - Fibrinogen-binding properties of the human platelet glycoprotein IIb-=IIIa complex: a study using crossed-radioimmunoelectrophoresis. PMID- 6286014 TI - Changes in cell surface antigen expression during hemopoietic differentiation. AB - Human bone marrow cells were separated on a fluorescence activated cell sorter (FACS) according to their binding of a series of monoclonal antibodies; the positive and negative fractions were cloned for erythroid burst and colony forming units (BFU-E and CFU-E) and myeloid colony-forming units (CFU-GM), and cytocentrifuge slides were prepared for microscopy of maturing precursors. The pattern of antigen expression on hemopoietic progenitor and precursor populations has been established using antibodies defining blood group (A, I/i), HLA associated (*A, B, C, DR, DC1), lineage specific, and transferrin receptor antigens. Like monomorphic HLA-DR, the antigen defined by monoclonal antibody OKT10 is expressed on the earliest progenitors and lost during differentiation, suggesting a role in interactions regulating the differentiation of these cells. The HLA-linked DC1 determinant, in contrast to HLA-DR, is not expressed at a detectable level on progenitor cells. Although a lineage-specific early antigen has not been identified, the transferrin receptor is expressed on the majority of erythroid progenitors, but only weakly on myeloid progenitors, and may provide an approach to isolating erythroid progenitors. These and earlier studies with monoclonal antibodies against HLA-DR and glycophorin now provide a detailed "map" of antigen expression during hemopoietic differentiation. PMID- 6286015 TI - Platelet-type von Willebrand's disease: characterization of a new bleeding disorder. AB - An autosomally transmitted bleeding diathesis sharing some, but not all, features previously described in von Willebrand's disease (vWd) was studied in five patients representing three generations of a single family. Bleeding times in the upper normal range in conjunction with low-normal platelet counts, normal factor VIII coagulant activity and VIII-related antigen, decreased VIII-ristocetin cofactor activity, selective decrease of the higher molecule weight factor VIII/von Willebrand factor (VIII/vWF) multimers, and increased ristocetin-induced platelet agglutination at low ristocetin concentrations were characteristic. Binding of patient VIII/vWF to washed normal platelets was within normal limits, whereas binding of normal VIII/vWF to patient platelets was significantly increased (p less than 0.001 at 0.6 mg/ml ristocetin). This disorder accordingly appears to involve an intrinsic platelet abnormality affecting platelet-VIII/vWF interactions. It is proposed that the concept of vWD be broadened to include patients with this abnormality, which may appropriately be called "Platelet-type von Willebrand's disease." PMID- 6286016 TI - [Laboratory diagnosis of infectious bursal disease (IBD) in northern Nigeria (1975-1979)]. PMID- 6286017 TI - Bovine leukosis in Nigeria--a preliminary study. PMID- 6286018 TI - A serological survey of bluetongue in Mozambique. PMID- 6286019 TI - [Demonstration of local production of antiherpetic antibodies in tears and its development over the years]. PMID- 6286020 TI - [Conjunctivitis caused by adenovirus: epidemiologic and therapeutic aspects]. PMID- 6286021 TI - Evidence for an A2/Ra adenosine receptor in the guinea-pig trachea. AB - 1 An attempt was made to determine whether the extracellular adenosine receptor that mediates relaxation in the guinea-pig trachea is of the A(1)/R(i) or A(2)/R(a) subtype.2 Dose-response curves to adenosine and a number of 5'- and N(6)-substituted analogues were constructed for the isolated guinea-pig trachea, contracted with carbachol.3 The 5'-substituted analogues of adenosine were the most potent compounds tested, the order of potency being 5'-N cyclopropylcarboxamide adenosine (NCPCA) > 5'-N-ethylcarboxamide adenosine (NECA) > 2-chloroadenosine > L-N(6)-phenylisopropyladenosine (L-PIA) > adenosine > D N(6)-phenylisopropyladenosine (D-PIA).4 The difference in potency between the stereoisomers D- and L-PIA on the isolated trachea was at the most five fold.5 Responses to low doses of adenosine and its analogues were attenuated after treatment with either theophylline or 8-phenyltheophylline. The responses to 2 chloroadenosine were affected to a lesser extent than were those to the other purines.6 Adenosine transport inhibitors, dipyridamole and dilazep, potentiated responses to adenosine, did not affect those to NCPCA, NECA, L-PIA and D-PIA but significantly reduced the responses to high doses of 2-chloroadenosine.7 Relaxations evoked by 9-beta-D-xylofuranosyladenosine which can activate intracellular but not extracellular adenosine receptors, were attenuated by dipyridamole but unaffected by 8-phenyltheophylline.8 The results support the existence of an extracellular A(2)/R(a) subtype of adenosine receptor and an intracellular purine-sensitive site, both of which mediate relaxation. PMID- 6286023 TI - Antagonism of the thromboxane-sensitive contractile systems of the rabbit aorta, dog saphenous vein and guinea-pig trachea. AB - 1 The thromboxane-sensitive contractile systems in spirally-cut preparations of the rabbit aorta, dog saphenous vein and guinea-pig trachea have been compared. The full or partial agonist activities of a range of bicyclic ring analogues were found to be remarkably similar on the three preparations. In addition, EP 045, a prostanoid with a phenylsemicarbazone omega-chain, blocked the action of both thromboxane A(2) (TXA(2)) and the bicyclic ring analogues. Using 11,9 epoxymethano prostaglandin H(2) as the agonist, linear Schild plots with slopes close to unity were obtained on each preparation; this suggests a competitive type of antagonism.2 Analogues of prostaglandin D(2) (PGD(2)), PGE(2) and PGF(2alpha) also contracted the three smooth muscle preparations; those analogues containing a 16-p-halophenoxy residue were highly active. On the rabbit aorta, EP 045 completely blocked the contractile actions of these agonists, perhaps indicating a single type of prostanoid receptor in this tissue. On the dog saphenous vein PGD(2), PGE(2) and 15-methyl PGE(2) exhibited relaxant activity when the tissue was partially contracted with either a thromboxane agonist or noradrenaline. On the guinea-pig trachea 16,16-dimethyl PGE(2) and the 16-p chlorophenoxy analogue of PGE(2) were potent contractile agents whose action was not blocked by EP 045. PGE(2) and 15-methyl PGE(2) showed similar properties but exhibited relaxant activity with increasing concentrations in the organ bath. Our results indicate the presence of three types of prostanoid receptors in the guinea-pig trachea: thromboxane- and PGE-sensitive systems mediating contraction and a PGE-sensitive system mediating relaxation.3 The similarity of the thromboxane-sensitive systems in the three smooth muscle preparations is discussed with particular reference to the differences in the equilibrium dissociation constants for EP 045. PMID- 6286022 TI - Reversible effects of tetanus toxin on striatal-evoked responses and [3H]-gamma aminobutyric acid release in the rat substantia nigra. AB - 1 The effects of sublethal doses of tetanus toxin on gamma-aminobutyric acid (GABA)-mediated synaptic transmission and [3H]-GABA release were studied in the rat substantia nigra. 2 Intranigral injections of tetanus toxin at 1-5 times the mouse LD50 dose produced ipsiversive circling behaviour which was maximal after 1 week and lasted 2-3 weeks. Rats then displayed normal behavior suggesting that the effects of the toxin were fully reversible. 3 In the treated nigra of circling rats there was a reduction in the striatal-evoked inhibition of compacta and reticulata neurones, but no change in their spontaneous firing rates. Some forms of striatal-evoked excitation were also reduced. Once rats had recovered from circling no alterations in the synaptic responses were detected. 4 In circling rats there were no differences in the sensitivities of neurones in the treated and untreated nigra to GABA or to other inhibitory neurotransmitters. 5 The Ca2+-dependent, K+-evoked release of [3H]-GABA from slices prepared from the treated nigra of circling rats was less than that from the untreated nigra of circling rats. No differences in nigral [3H]-GABA release were observed once rats had recovered from the circling behaviour. 6 The results demonstrate that doses of tetanus toxin which produce reversible behavioural effects can interfere reversibly with GABA-mediated synaptic transmission by a presynaptic mechanism which probably involves a reduction in transmitter release. PMID- 6286024 TI - Drugs that increase gamma-aminobutyric acid transmission protect against the high pressure neurological syndrome. AB - 1 The effects on the high pressure neurological syndrome (HPNS) of drugs which facilitate gamma-aminobutyric acid (GABA) transmission were investigated. Threshold pressures for the onset of the behavioural signs of the HPNS in mice- tremors and convulsions were established. 2 Flurazepam hydrochloride 20 and 10 mg/kg and sodium valproate 800 and 400 mg/kg substantially raised the threshold pressures for both tremor and convulsions. 3 Amino-oxyacetic acid 35 and 25 mg/kg and diaminobutyric acid 600 mg/kg also significantly increased the thresholds. Muscimol 1 mg/kg (and 150 ng i.c.v.) was ineffective at non-toxic doses. 4 These effects paralleled the drugs' ability to raise the convulsion threshold to intravenous infusion of bicuculline in mice. 5 These results demonstrate that drugs with actions more selective than those of the general anaesthetics are effective against the HPNS. It is also possible that there is a GABAergic component to the effects of general anaesthetics on the HPNS. PMID- 6286025 TI - Effects of ritodrine, a beta 2-adrenoceptor agonist, on smooth muscle cells of the myometrium of pregnant rats. AB - 1 Effects of ritodrine on the electrical and mechanical activities of the myometrium of pregnant rats were investigated in relation to cyclic adenosine 3',5'-monophosphate (cyclic AMP) production and the actions of isoprenaline. 2 In the longitudinal muscle cells of the 22nd day of gestation, ritodrine (greater than 10(-8) M) hyperpolarized the membrane, reduced the membrane resistance and suppressed the amplitude of contraction. Increased concentrations of ritodrine (greater than 10(-7) M) suppressed the generation of spikes and increased the amount of cyclic AMP produced. These actions of ritodrine were much the same as those of isoprenaline but the dose-response curves of all parameters were shifted to the right. 3 The effects of ritodrine on the circular muscle cells at the 22nd day of gestation were similar to those observed in the longitudinal muscle cells. 4 In the circular muscle cells at the 17-19th days of gestation, ritodrine produced neither a hyperpolarization nor an increase in cyclic AMP production (up to 10(-5) M) but did reduce the plateau potential and did relax the tissue (greater than 10(-7) M). 5 In the longitudinal muscle cells, during the 17-19th days of gestation, the responses of the tissues were similar to those observed in circular muscles on the 22nd day of gestation. These actions of ritodrine were suppressed by propranolol (10(-6) M). 6 These results indicate that ritodrine is a beta-adrenoceptor agonist and its mechanism of action similar to that of isoprenaline. However, the potency of ritodrine was approximately 100 times less than that of isoprenaline. PMID- 6286026 TI - Selective depression of synaptic transmission of spinal neurones in the cat by a new centrally acting muscle relaxant, 5-chloro-4-(2-imidazolin-2-yl-amino)-2, 1, 3-benzothiodazole (DS103-282). AB - 1 The effects of a new muscle relaxant, 5-chloro-(2-imidazolin-2-yl-amino)-2, 1, 3-benzothiodazole (DS103-282) have been examined on segmental reflexes and responses of single neurones in the spinal cord of the anaesthetized cat to stimulation of peripheral afferents, ventral roots, acetylcholine and various amino acids. Drugs were administered intravenously and/or iontophoretically.2 Polysynaptic reflexes were depressed in a dose-dependent manner by 0.01-0.1 mg/kg DS103-282 whereas monosynaptic reflexes were relatively insensitive to this agent.3 In studies on single dorsal horn neurones, iontophoretically and intravenously administered DS103-282 depressed synaptic excitatory responses, polysynaptic responses being much more sensitive to this agent than monosynaptic responses. In contrast (-)-baclofen preferentially reduced monosynaptic excitation.4 Doses or ejecting currents of DS103-282 which greatly depressed polysynaptic excitatory responses also reduced spontaneous firing of neurones, but either had no effect or minimal depressant effects on responses to iotophoretically administered excitant amino acids. Acetylcholine-induced excitation of Renshaw cells was depressed by iontophoretically (but not intravenously) administered DS103-282, although ventral root-evoked responses of these cells were insensitive to this agent.5 Inhibition of spinal neurones by stimulation of peripheral nerves or by iontophoresis of gamma-aminobutyric acid or glycine was unaffected by DS103-282.6 These results indicate that DS103-282 preferentially depresses peripherally evoked polysynaptic excitation of spinal neurones probably by an action on the terminals of excitatory interneurones. PMID- 6286027 TI - Naloxone inhibits early arrhythmias resulting from acute coronary ligation. AB - The intravenous administration of naloxone 15 min before acute coronary artery ligation in both anesthetized and conscious male rats markedly reduced the incidence and severity of the ventricular arrhythmias that occur within 30 min of the onset of myocardial ischaemia. The incidence of ventricular fibrillation was especially reduced and, in conscious rats, the survival 16 h after ligation was increased from 27% (in the controls) to 58 and 73% after 2 and 4 mg/kg naloxone respectively. One possible explanation of these results implies a detrimental effect of released endorphin in the early stages of myocardial ischaemia. PMID- 6286028 TI - A comparative study of beta-adrenoceptors in human and porcine lung parenchyma strip. AB - 1. Responses to (+/-)-isoprenaline (Iso), (-)-adrenaline (Adr) and (-) noradrenaline (NA) were compared in isolated preparations of human and porcine lung parenchyma strip. 2. The order of relaxant potencies of these catecholamines in both human and porcine lung parenchyma was Iso greater than Adr greater than NA (1:0.24:0.01, human; 1:0.21:0.01.pig). These results suggest that beta 2 adrenoceptors predominate in both types of lung parenchyma strip. 3. pA2 values for the beta-adrenoceptor antagonist, propranolol (non-selective), with Iso as the agonist, in human and porcine lung strips were 7.84 and 7.83 respectively and for atenolol were 6.50 and 5.35 respectively. Taken as a whole results indicate the existence of an apparently homogeneous population of beta 2-adrenoceptors in porcine parenchyma strip, while both beta 1 and beta 2-adrenoceptors were revealed in human lung parenchyma. PMID- 6286030 TI - The resistance of polyglycolic acid sutures to attack by infected human urine. AB - Polyglycolic acid sutures were incubated at 37 degrees C in broth, and in 0.22 mu millipore filtered human urine infected with Escherichia coli, Str. faecalis, Proteus mirabilis and Pseudomonas aeruginosa. At 3 days the Proteus had destroyed the polyglycolic acid sutures in the urine but not in the broth. None of the other organisms had any effect on the tensile strength of the suture material. Under the scanning electron microscope the Proteus-destroyed polyglycolic acid sutures showed multiple transverse microfractures. Polyglycolic acid sutures should not be used for closing urothelium in patients with a proven Proteus infection. PMID- 6286032 TI - Collapse of cervical spine treated by Down's Ace mark III halo assembly. PMID- 6286029 TI - Release of beta-lipotropin- and beta-endorphin-like material induced by angiotensin in the conscious rat. AB - 1 The influence of the renin-angiotensin system on plasma beta-endorphin-like immunoreactivity (beta-EI) was investigated in the conscious rat by use of a radioimmunoassay for beta-endorphin without prior extraction.2 Intravenous infusion of angiotensin I, II or (des-1-Asp)angiotensin II (angiotensin III) caused a dose-dependent increase in plasma beta-EI, angiotensin III infusion being less effective than angiotensin I or II. The plasma adrenocorticotrophin (ACTH) levels too were elevated by angiotensin II. The receptor antagonist, saralasin, prevented the angiotensin II-induced beta-EI release as did dexamethasone pretreatment.3 Both the release of beta-EI and the pressor response to angiotensin I were abolished by the converting enzyme inhibitor, captopril (SQ 14225). In contrast, captopril did not affect the action of angiotensin II.4 In view of the appreciable cross-reactivity of beta-lipotropin (beta-LPH) in our assay, plasma beta-EI was analysed by Sephadex G-50 chromatography. In plasma extracts of angiotensin II-infused rats, immunoreactivity corresponding to human beta-endorphin comprised about 49% of the total immunoreactivity, whereas 51% co migrated with human beta-LPH.5 The increase in plasma levels of beta-EI elicited by angiotensin II was diminished by about 35% in rats with a hereditary absolute lack of vasopressin (Brattleboro rats), when compared to normal rats.6 These results suggest that the renin-angiotensin system can stimulate the secretion of beta-LPH and beta-endorphin with ACTH from rat anterior pituitary. One link in mediating the response appears to be vasopressin. The physiological function remains to be defined. PMID- 6286031 TI - An assessment of dynamic gastric scanning with 99Tcm-pertechnetate. AB - Following intravenous injection, technetium-99m (99Tcm) is selectively taken up by the stomach, and this uptake, which is stimulated by pentagastrin, may be quantified using a gamma camera. Using this property of 99Tcm as a means of assessing gastric function, we found that the mean rate of 99Tcm uptake was significantly greater in 24 duodenal ulcer patients than in 10 healthy controls (P less than 0.01). Scanning values did not correlate with BAO of MAO which were measured on a separate day. Both vagotomy (n = 10) and cimetidine infusion (n = 5) significantly reduced the rate of uptake of 99Tcm (P less than 0.01). Thus, although 99Tcm reflects a function of the stomach which is similar to acid secretion, scanning did not give a good estimate of acid secretion in this group of subjects. PMID- 6286033 TI - Ranitidine in acute upper gastrointestinal haemorrhage. PMID- 6286035 TI - Manual motor performance in a deafferented man. AB - We have studied manual motor function in a man deafferented by a severe peripheral sensory neuropathy. Motor power was almost unaffected. Our patients could produce a very wide range of preprogrammed finger movements with remarkable accuracy, involving complex muscle synergies of the hand and forearm muscles. He could perform individual finger movements and outline figures in the air with high eyes closed. He had normal pre- and postmovement EEG potentials, and showed the normal bi/triphasic pattern of muscle activation in agonist and antagonist muscles during fast limb movements. He could also move his thumb accurately through three different distances at three different speeds, and could produce three different levels of force at his thumb pad when required. Although he could not judge the weights of objects placed in his hands without vision, he was able to match forces applied by the experimenter to the pad of each thumb if he was given a minimal indication of thumb movement. Despite his success with these laboratory tasks, his hands were relatively useless to him in daily life. He was unable to grasp a pen and write, to fasten his shirt buttons or to hold a cup in one hand. Part of hist difficulty lay in the absence of any automatic reflex correction in his voluntary movements, and also to an inability to sustain constant levels of muscle contraction without visual feedback over periods of more than one or two seconds. He was also unable to maintain long sequences of simple motor programmes without vision. PMID- 6286034 TI - Comparative in-vitro activity of selected new beta-lactam antimicrobials against Neisseria gonorrhoeae. AB - Four new beta-lactam antimicrobials, ceftriaxone, cefotiam, cefonicid, and mecillinam, were evaluated in vitro against 72 beta-lactamase-negative and 26 beta-lactamase-positive isolates of Neisseria gonorrhoeae. Ceftriaxone was the most active of the antimicrobials tested. It inhibited all isolates, regardless of beta-lactamase activity, at a concentration of less than or equal to 0.015 microgram/ml. Cefotiam and cefonicid were also active against both groups but not as active as ceftriaxone. Both groups of N gonorrhoeae showed a high degree of resistance against mecillinam. PMID- 6286037 TI - Electrical signs of an oligosynaptic visual projection to the rat hippocampus. AB - In rats with pons transection photic or optic nerve stimulation elicits a response in dorsal hippocampus with approximately with same latency, amplitude and time course as the response in striate cortex. After optic nerve stimulation a positive polyphasic deflection with the last peak at 10 ms is followed by a larger biphasic major deflection with peaks at 15 and 25 ms and later, slower deflections at intervals of 125 ms. The polyphasic deflection is maximal in the inferior part of the hippocampus but does not reverse polarity; the other deflections reverse above and below an isoelectric point in the hilum of the dentate gyrus, a distribution attributable to depolarization in the molecular layers, perhaps also cell bodies, of that structure. The major deflection is more sensitive to changes in optic nerve stimulus strength than the response in striate cortex and is more resistant to reduction in amplitude during repetitive stimulation, following frequencies up to 50/s. The pathway between retina and hippocampus for all parts of the response is interrupted by lesions in the tectum. The major deflection is abolished by lesions in the posterior cingulum. In the posterior cingulum the pathway has fast and slow components in the lower and upper portions, respectively, associated with the first and second parts, respectively, of the major deflection. The pathway is not interrupted by lesions in the fornix, septal nuclei, anterior cingulum, anterior medial thalamus or medial midbrain ventral to the superficial tectum. There are complex interactions, up to several hundred milliseconds in duration, between the response to optic nerve stimulation and those elicited by stimulation in the cingulum, midbrain and thalamus. Tonic influences on the dentate gyrus from cingulum and tectum are described. PMID- 6286036 TI - Methacholine pupillary responses in third nerve palsy and Adie's syndrome. AB - In order to examine the clinical usefulness of methacholine in assessing the site of ocular parasympathetic lesions, pupillary responses in man were measured in postganglionic (Adie's syndrome) and preganglionic third nerve lesions involving the pupil and in controls. From previous work with methacholine it might have been expected that greater constriction would occur in the postganglionic lesions but similar responses were found in both. Corneal hypoxia due to ptosis appeared unlikely to affect corneal permeability significantly and it is probable that these results reflect an increase responsiveness of the iris at, or distal to, the site of muscarinic acetylcholine receptors. Pupils contralateral to third nerve palsy, when tested on a separate occasion, also constricted by an amount approximately proportional to that of the clinically abnormal pupil. The possibilities that this may result in some way from reduction in total retinal illumination, or from retrograde changes in preganglionic pupilloconstrictor neurons affecting contralateral pupilloconstrictor neurons via central pathways, are discussed. It is concluded that supersensitivity to methacholine, tested carefully in the manner described, is a useful guide to the presence of parasympathetic denervation or decentralization, but that it is not reliable in distinguishing between the two sites. PMID- 6286038 TI - Effects of short-term exposure to catecholestrogens on catecholamine turnover in the preoptic-hypothalamic brain of ovariectomized rats. AB - The effects of a series of catecholestrogens (2-hydroxyestrogens and 4 hydroxyestrogens) were compared to those of a primary estrogen, ethynylestradiol (EE2), on the catecholaminergic system in the preoptic-hypothalamic rat brain. Adult ovariectomized rats received single injections (100 microgram s.c.) of EE2, 2-hydroxyestradiol (2-OHE2), 2-hydroxyethynylestradiol (2-OHEE2), 4 hydroxyestradiol (4-OHE2) or 4-hydroxyethynylestradiol (4-OHEE2). Eight hours after estrogen administration the animals were killed, and the concentrations of dopamine (DA), noradrenaline (NA) and adrenaline (A) were determined radioenzymatically in the preoptic area (POA) and the mediobasal hypothalamus (MBH). EE2 and all the catecholestrogens tested uniformly suppressed (P less than 0.01) serum LH levels. This correlated well with the decreased turnover rate of A in the POA. In both the POA and the MBH of EE2-treated animals the turnover rate of NA was markedly decreased whereas the concentrations of catecholamines remained unaffected. The catecholethynylestrogens, potent inhibitors of catechol O-methyltransferase, caused two-fold increases of NA concentrations in the POA (2 OHEE2 and 4-OHEE2) and MBH (2-OHEE2) without affecting the turnover rate. Thus, although EE2 and all the catecholestrogens tested uniformly suppressed LH release they induced highly different effects on the noradrenergic system In the preoptic hypothalamic brain. PMID- 6286039 TI - Clorgyline delays the phase-position of circadian neurotransmitter receptor rhythms. AB - The number of alpha- and beta-adrenergic, muscarinic cholinergic, opiate, and benzodiazepine receptors in rat forebrain, and dopamine and benzodiazepine receptors in striatum, change throughout the day. The diurnal rhythms of these receptors were altered by treatment with the monoamine-oxidase inhibitor clorgyline: following treatment some or all rhythm characteristics of wave form, amplitude, 24-h mean, and phase, were affected. One common effect of treatment was a delay in phase-position of binding to alpha- and beta-adrenergic, opiate and benzodiazepine receptors. Additionally, the nocturnal elevation in pineal melatonin which normally returns to baseline at light onset, persisted 3 h into the light period after clorgyline administration. These biochemical observations extend behavioural findings that clorgyline can delay the phase-position of rodent nocturnal activity onset, and does so by slowing the central circadian pacemaker. PMID- 6286040 TI - A role for striatal beta-adrenergic receptors in the regulation of dopamine release. PMID- 6286041 TI - Localization of opiate and histamine H1-receptors in the primate sensory ganglia and spinal cord. AB - The autoradiographic localizations of opiate and histamine H1-receptors were studied quantitatively and compared following dorsal root section in the primate spinal cord. High densities of opiate and H1-receptors were found in the superficial layers of the dorsal horn. Dorsal root section produced a 40-50% loss of both receptors types throughout layers I and II with no loss in deeper layers of the dorsal horn. "Bursts' of opiate and H1-receptor labelling was observed over a population of small diameter cells (35-45 micron or micrometer) in the primate dorsal root ganglion. These cells represented approximately 8% of the total cells present. Serial consecutive sections demonstrated a population of cells bearing both opiate and histamine H1-receptors. PMID- 6286042 TI - Studies on the intrathecal effect of beta-endorphin in primate. AB - The intrathecal administration of beta-endorphin in the primate through an indwelling spinal catheter, produced a significant elevation in the nociceptive threshold as measured by the discrete trial shock titration task. The time of onset, duration of effect and magnitude of effect were all dose-dependent over a range of 150-750 micrograms. The effects were antagonized in a dose-dependent fashion by the systemic administration of naloxone. Aside from the elevations in the shock titration threshold produced by intrathecal beta-endorphin, no untoward effects on the animal's motor function or behavioral reactivity was noted. Significantly, unlike morphine, intrathecal beta-endorphin failed to produce any signs of scratching behavior at the doses used in these experiments. Once daily administration of intrathecal beta-endorphin (500 micrograms) showed a significant progressive decline in the antinociceptive effect over an 8-day period. Animals made tolerant to beta-endorphin in this fashion showed a significantly reduced response to an otherwise active dose of intrathecal morphine, indicating evidence for cross tolerance. PMID- 6286043 TI - Flurazepam interaction with sodium and potassium channels in squid giant axon. AB - External application of 0.05-1.0 mM flurazepam was found to partially block both sodium and potassium currents in voltage-clamped squid giant axons. At the same concentration the fractional block of the potassium current was found to be 3 times greater than that of the sodium current. In the presence of the drug the potassium current appeared to "inactivate', as flurazepam block became more profound during the course of the depolarization. The decay of the potassium current can be explained by a model in which flurazepam enters and blocks the potassium channels only after they have opened. Once bound in the potassium channel, removal of flurazepam from its binding site develops slowly (tau = 48 ms). Thus repetitive stimulation of the nerve produced a cumulative block. When applied inside the axon flurazepam was found to be 1.5 (n = 4) times more potent blocker of potassium channels than following external application. This result suggests that when applied externally, a neutral form of the drug diffuses across the membrane and blocks occurs from the inner end of the channel. PMID- 6286044 TI - Rapid appearance of intraventricularly administered neuropeptides in the peripheral circulation. AB - 125I-Labeled cholecystokinin octapeptide ([125I]CCK-OP) diffuses rapidly by a non carrier-mediated mechanism into the peripheral blood following injection into the lateral ventricle of a rabbit. In contrast, no [125I]CCK-OP was observed in the CSF following intravenous injection. This unidirectional free transport mechanism from CSF to blood may apply to other neuropeptides as well. PMID- 6286045 TI - Opiate radioreceptor assay distinguishes between anesthetics and convulsants: the effect of pH. AB - The specific binding of [3H]naloxone to brain tissue is perturbed by anesthetics and convulsants in a rank order of potency related to lipophilicity of the drugs. Neurodepressants can be distinguished from neuroexcitants in the assay, because sodium ion enhances the action of the former and antagonizes the action of the latter. As the pH of the assay medium is varied from 6.7 to 7.7 to 8.4, perturbation of [3H]naloxone binding by the neurodepressants, diethyl ether and Isoindoklon, is unaffected either in the presence or absence of sodium. In contrast, the antagonism by sodium of the neuroexcitants, DDT and Indoklon (the convulsant isomer of Isoindoklon), is progressively enhanced with increasing pH. These observations on the effects of pH extend earlier ones on ion effects and should contribute to the delineation of specific membrane loci that mediate the actions of neurodepressants as distinct from sites that mediate the actions of neuroexcitants. PMID- 6286046 TI - The relationship between alpha 2-adrenoceptor selectivity and anticonvulsant effect in a series of clonidine-like drugs. AB - Clonidine and the 4 clonidine-like drugs: 44-549, lofexidine, guanfacine, and CP 14,304-18, had anticonvulsant activity against pentylenetetrazole-induced convulsions in rats. The magnitude and dose range over which anticonvulsant effects were observed was related to the selectivity of the compounds for alpha 2 and alpha 1 adrenoceptors. Selective alpha 2 adrenoceptor agonists may form a new group of anticonvulsants. PMID- 6286047 TI - The influence of selective adeno- and neurointermedio-hypophysectomy upon plasma and brain levels of beta-endorphin and their response to stress in rats. AB - Selective ablation of the anterior lobe (AL) of the pituitary led to a fall in basal plasma levels of beta-endorphin immunoreactivity (beta-EI) at 3 and 20 weeks post-surgery (p.s.). Further, the stress-evoked rise in circulating levels of beta-EI was abolished. This operation did, however, severely deplete the beta EI content of the neurointermediate lobe (NIL). Removal of the NIL did not, in contrast, decrease the beta-EI content of the AL but depressed basal plasma levels of beta-EI at 3 weeks p.s. and attenuated, but did not abolish, the increase in these elicited by stress at both 3 and 20 weeks p.s. In rats not possessing a NIL, a secretion of beta-EI into plasma can thus occur. The possibility that NIL pools of beta-EI contribute to circulating levels of beta-EI is discussed. Removal of the AL depressed the beta-EI content of the hypothalamus and periventricular tissue at 3 and 20 weeks p.s. The Met-enkephalin immunoreactivity (ME-I) content of the hypothalamus was, in contrast, unaffected. These animals still responded to stress at 20 weeks p.s. with a significant fall in hypothalamic levels of beta-EI. Extirpation of the NIL did not, in contrast, change brain levels of either beta-EI or ME-I. The presence of the AL, but not the NIL, is thus essential for the maintenance of usual levels of beta-EI and ME I in the brain. PMID- 6286048 TI - Site of action potential generation in a giant neuron of Aplysia californica. PMID- 6286049 TI - Cytochemical localization of ouabain-sensitive, K+-dependent p-nitro phenylphosphatase (transport ATPase) in the mouse central and peripheral nervous systems. AB - Enzyme activity, representing the sites of K+-dependent p-nitrophenylphosphatase (K+-pNPPase), a component of the transport adenosine triphosphatase (Na+,K+ ATPase) system, has been localized at the ultrastructural in both cerebral cortex and in sciatic nerve of the mouse. Normal mice and animals with mechanically injured blood-brain barrier (BBB) were used. In the cerebral cortex, positive reaction was found in synapses, plasmalemma of neurites (axons and dendrites), in endothelial cell of microblood vessels and in the plasmalemma of mesothelial cells of the pia mater. In the sciatic nerve, a strong reaction was present in the nodes of Ranvier, with weaker reaction in the internodal areas of the axolemma. In the endothelial cells of normal blood vessels, the reaction product was localized on the luminal and abluminal, or only on the abluminal plasmalemma. After damage of BBB, numerous invaginations, pits and pinocytic vesicles showing positive reaction in their limiting membrane appeared in the endothelial cells. PMID- 6286050 TI - Identification of gK systems activated by [Ca2+]. AB - Rhythmic caffeine hyperpolarizations generated in bullfrog sympathetic ganglion cells are assumed to be caused by periodic increase in gK due to rise in [Ca2+]i7 -9,13. Caffeine-induced outward currents seem to be composed of two different components, which show different pharmacological natures and also different dependencies on membrane potential changes. These two components may be generated by activation of two voltage-dependent K+ currents, namely IK1 (the delayed rectifier K+ current) and IK2 (IM) of ganglion cells. These results suggested that at least two different gK systems were activated by [Ca2+]i in sympathetic ganglion cells. PMID- 6286051 TI - Differential effects of chronic morphine and naloxone on opiate receptors, monoamines, and morphine-induced behaviors in preweanling rats. AB - Rats were administered either chronic morphine, naloxone or saline from 1 to 21 days of age. At 22 days of age, animals were sacrificed and various CNS areas were assayed for specific binding of [3H]naloxone and steady-state levels of norepinephrine, dopamine and serotonin, as well as turnover of norepinephrine and dopamine. In addition, some animals at 22 days of age were assessed for morphine induced changes in activity, hot-plate paw-lick latency, and rectal body temperature. Chronic naloxone treatment produced an increase in the number of ligand binding sites in hypothalamus, striatum and cortex, but did not alter monoamine systems or the efficacy of morphine. In contrast, chronic morphine treatment produced tolerance to the hypoactive and antinociceptive effects of morphine, but did not alter ligand binding or monoamine systems. These results demonstrate that developing opiate receptor systems in brain are more responsive to chronic receptor blockade than to chronic receptor activation and that an alteration in the development of opiate receptor systems does not necessarily produce a concomitant alteration in either monoamine systems or the behavioral efficacy of morphine. PMID- 6286052 TI - An evaluation of adrenal mass and adrenal cholesterol as measures of adrenal activity. AB - 1. The effects of twice daily injections of corticotrophin (1 IU/kg body weight) or restriction of food intake to 75% of normal on body mass, adrenal mass and adrenal cholesterol were determined on chicks from 1 to 21 d of age. 2. Only the birds subjected to restricted feeding showed a reduced growth rate. 3. There was no adrenal hypertrophy in birds receiving corticotrophin but in the restricted group there was transient hypertrophy at 2 weeks. 4. Depletion of adrenal cholesterol was noted only in the birds receiving corticotrophin. 5. It is concluded that neither depletion of cholesterol nor hypertrophy is an inevitable consequence of enhanced adrenal cortical activity. PMID- 6286053 TI - [Detection and mapping of conserved nucleotidic sequences between the genomes of human papillomavirus 1 a and bovine papillomavirus 1 by electron microscope heteroduplex analysis (author's transl)]. AB - Three regions of partial homology have been detected between the genomes of human papillomavirus 1 a (HPV-1 a) and bovine papillomavirus 1 (BPV-1) by electron microscope analysis of heteroduplex molecules. These regions contain about 25% of non-homologous bases and represent 13% of the genome length. They have been mapped on the viral DNAs. This allowed a reciprocal orientation of the physical maps of the two genomes. Two regions are located in the middle of the transforming fragment of BPV-1 DNA and the third, at at diametrically opposite position, falls into the region of the genome coding most probably for a viral structural protein. PMID- 6286054 TI - Gestational trophoblastic neoplasia. AB - As a result of modern diagnostic and therapeutic techniques, nearly 100 percent of patients with GTN can be cured of a disease thay only a few years ago had a high death rate. To achieve this, skillful mixing of chemotherapy, surgery, and irradiation, as well as supportive therapy, are necessary. Understanding and utilizing the HCG assay to monitor disease status is vital. With this approach, essentially all of these patients should survive, many to have successful future pregnancies. PMID- 6286055 TI - Neurofibrosarcoma at irradiation site in a patient with neurofibromatosis and Wilms' tumor. AB - A female patient with neurofibromatosis had nephrectomy performed because of Wilms' tumor at the age of four and a half. She received radiation therapy and chemotherapy (actinomycin D) after surgery. She had subsequent local recurrence and lung metastasis, which were surgically excised and successfully treated with additional radiation therapy and chemotherapy (vincristine and actinomycin D). However, neurofibrosarcoma at the irradiation site developed seven years after radiation therapy. She died 22 months later because of recurrence and metastasis of neurofibrosarcoma. Radiation therapy's association with malignant transformation of neurofibroma is discussed. PMID- 6286056 TI - Hodgkin's disease and infectious mononucleosis: is there a causal association? PMID- 6286057 TI - Taking laetrile: conversion to medial deviance. AB - For members of the CCS, laetrile use occurs in a self-help social context where users derive substantial social and emotional support from fellow members. We are convinced that much of the appeal of laetrile results from this social context in which it is frequently used. The CCS and the extensive network of similar organizations are fulfilling unmet needs in cancer patients and those fearful of cancer. Until conventional cancer therapy deals more successfully with the social and psychological needs of cancer patients, promoters of laetrile will till fertile ground. PMID- 6286058 TI - [Isolation and identification of two reovirus strains (author's transl)]. PMID- 6286059 TI - [Detection of IgA serum antibodies to EB virus VCA in patients having survived nasopharyngeal cancer for over 15 years (author's transl)]. PMID- 6286060 TI - [Human cytomegalovirus and disease (author's transl)]. PMID- 6286061 TI - Inhibition of parathyroid hormone-stimulated bone resorption by monovalent cation ionophores. PMID- 6286062 TI - Biochemical characterization of 1,25(OH)2D3 receptors in chick embryonal duodenal cytosol. AB - This study presents measurements of serum vitamin D metabolites, calcium and phosphorus as well as measurements of the equilibrium dissociation constant for duodenal 1,25(OH)2D3 receptor in 15-, 18-, 19-, and 20-day chick embryos in comparison to that in 1- and 118-day-old chicks and to vitamin D-deficient chicks. The present results showed that: (a) serum 1,25(OH)2D and 24,25(OH)2D levels rise from 15 and 18 to days 19 and 20 of embryonic development while serum phosphate levels are stable; (b) serum calcium levels rise at hatching to adult levels; (c) the duodenal 1,25(OH)2D3 receptor is detectable in 15-day-old embryo and has a Kd similar to that of 118-day-old vitamin D-replete chicks; and (d) the activity of 1,25(OH)2D3 receptor in chick duodenal cytosol is maximal at hatching. PMID- 6286063 TI - Suppressive effect of chronic glucocorticoid treatment on circulating calcitonin in man. PMID- 6286064 TI - The role of free calcium ion in calcium release in skinned muscle fibers. AB - Major questions in excitation--contraction coupling of fast skeletal muscle concern the mechanism of signal transmission between sarcolemma and sarcoplasmic reticulum (SR), the mechanism of SR Ca release, and operation of the SR active transport system during excitation. Intracellular Ca movement can be studied in skinned muscle fibers with more direct control, analysis of 45Ca flux, and simultaneous isometric force measurements. Ca release can be stimulated by bath Ca2+ itself, ionic "depolarization," Mg2+ reduction, or caffeine. The effectiveness of bath Ca2+ has suggested a possible role for Ca2+ in physiological release, but this response is difficult to analyze and evaluate. Related evidence emerged from analysis of other responses: with all agents studied, stimulation of 45Ca efflux is highly Ca2+-dependent. The presence of a Ca chelator prevents detectable stimulation by ionic "depolarization" or Mg2+ reduction and inhibits the potent caffeine stimulus; inhibition is graded with chelator concentration and caffeine concentration, and is synergistic with inhibition by increased Mg2+. The results indicate that a Ca2+-dependent pathway mediates most or all of stimulated 45Ca efflux in skinned fibers, and has properties compatible with a function in physiological Ca release. PMID- 6286065 TI - Roles of extracellular and "trigger" calcium ions in excitation--contraction coupling in skeletal muscle. AB - The experimental observations leading to the development of the "trigger" calcium hypothesis of excitation--contraction (E--C) coupling in skeletal muscle are discussed. Also considered in some detail are the experimental technique problems which interfere with the demonstration of this role for calcium. New findings reported are observations showing that in a zero Ca2+ solution after a delay of about 6--10 min, there is a stimulation of Ca2+ efflux. This is of sufficient size, even in very small toe muscles, to restore the twitch which previously had been reduced in size in the zero Ca2+. In studies with isolated fibre preparations it was demonstrated that depolarization contractures required extracellular Ca2+ ions for E--C coupling whereas twitches could use membrane bound "trigger" calcium ions. Thus in zero Ca2+ the contractures were eliminated in a few seconds but twitch elimination took a few minutes. Finally, the roles in E--C coupling played by "trigger" and extracellular Ca2+ ions are summarized and discussed. PMID- 6286066 TI - Extracellular divalent cations in excitation--contraction coupling: hypertonic solution effects. PMID- 6286067 TI - The role of calcium in excitation--contraction coupling in crustacean muscle fibers. AB - The evidence that calcium (Ca) plays an important role in electrical activity and an essential role in excitation--contraction (E--C) coupling in crustacean muscles is reviewed. These muscles produce graded electrical and mechanical responses to applied depolarizations. Removal of Ca from the bath solution eliminates both responses. Addition of Ba2+ or Sr2+ to Ca-free saline restores membrane electrogenesis, and all-or-none action potentials can be induced. With Sr2+ vigorous contractions are produced, whereas Ba action potentials evoke minimal or no tension, showing that rapid depolarization of the membrane potential is not sufficient per se for E--C coupling in crab and barnacle muscle. Several inorganic (e.g., multivalent cations) and organic (e.g., aminoglycoside antibiotics) which block membrane Ca channels block electrogenesis and contraction. However, the "Ca antagonists" verapamil and D600 also block Ca uptake at intracellular storage sites, resulting in spontaneous contractions and the delayed relaxation of small contractions associated with residual Ca currents. The evidence that the Ca which enters the fibres needs to release Ca from intracellular storage sites to produce contractions is detailed and discussed. Finally, a model for E--C coupling is discussed. This model includes the sites and mechanisms of action for several chemicals which modify E--C coupling in crustacean muscle fibres. PMID- 6286068 TI - Ca2+ coupling in vascular smooth muscle: Mg2+ and buffer effects on contractility and membrane Ca2+ movements. AB - An examination of the literature, over the past two decades, reveals that (1) in studies of different types of vascular smooth muscles, Mg2+ is often either left out of physiological salt solutions or reduced in concentration compared with that in blood; and (2) when excitation--contraction coupling processes have been examined in isolated vascular tissues and cells, a number of artificial (synthetic) amine and organic zwitterion buffers have often been substituted for the naturally occurring bicarbonate and phosphate anions found in the blood and in cells. The influence of extracellular magnesium ions ([Mg2+]0) on tone, contractility, reactivity, and divalent cation movements in vascular smooth muscles, and how they may relate to certain vascular disease states, is reviewed. Data are presented and reviewed which indicate that many of the most commonly used artificial buffers (e.g., Tris, HEPES, MOPS, Bicine, PIPES, imidazole) can exert adverse effects on contractility and reactivity of certain arterial and venous smooth muscles. The data reviewed herein suggest that [Mg2+]0 and membrane Mg are important in the regulation of vascular tone, vascular reactivity, and in control of Ca uptake, content, and distribution in smooth muscle cells. [HCO3-]0 and (or) PO4(2-) anions may be important for normal maintenance of excitability and reactivity and in the control of Ca uptake, content, and distribution in smooth muscle cells. PMID- 6286069 TI - Calcium and excitation--contraction coupling in vascular smooth muscles. AB - The roles of Ca2+ in excitation--contraction coupling in vascular smooth muscle have been difficult to delineate, primarily because unambiguous association of specific Ca2+ components with morphologically defined cellular structures could not be attained. More recent use of washouts in La3+-substituted solutions at low temperature (to remove superficial Ca2+ and retain cellular Ca2+), Scatchard coordinate plots (to identify incubation conditions appropriate for examining predominantly high or low affinity Ca2+ components), and high concentrations of Sr2+ (to remove high but not low affinity Ca2+) have facilitated qualitative and quantitative separation of different Ca2+ fractions. The release of high affinity Ca2+ elicited with norepinephrine and the increase in uptake of low affinity Ca2+ obtained with high K+ have been clearly demonstrated, and may directly measure or indirectly reflect changes in the level of intracellular free Ca2+. In other types of vascular smooth muscle (e.g., renal vessels, coronary arteries), similar Ca2+ components also appear to be present, but their relative size and functional importance for regulation of contractile responsiveness can differ. PMID- 6286070 TI - Calcium in excitation--contraction coupling of frog skeletal muscle. AB - A principal step in the process leading to muscle contraction is the intracellular release of Ca2+. We have detected and compared some physical and chemical events that reflect Ca2+ release in contracting frog skeletal muscle cells, described the effects of some agents that are believed to alter intracellular Ca2+ release during contraction, and speculated about the role of Ca2+ release in influencing some of the mechanical properties of frog muscle. The specific physical features recorded were changes in striation spacing, myofibrillar orientation, and force development. The chemical feature was the relative change in intracellular [Ca2+] recorded as light emission from cells microinjected with the Ca2+-sensitive protein aequorin. The presence or absence of a correlation among these variables has been used (i) to evaluate the action of some agents thought to change intracellular Ca2+ release in excitation- contraction (E--C) coupling, (ii) to further substantiate the effects of cell length on Ca2+ release, and (iii) to examine some details of models for E--C coupling. The results showed that potentiating agents enhance and prolong intracellular Ca2+ release without changing the rate of Ca2+ removal during E--C coupling. This extra Ca2+ does not produce the same effect on contractions at all lengths. Contractility is inversely related to cell length, and Ca2+-induced activation is normally less than maximum not only at short lengths but also at optimal striation spacings. PMID- 6286071 TI - Possible role of the transverse tubules in accumulating calcium released from the terminal cisternae by stimulation and drugs. AB - The size of the rapidly exchanging and slowly exchanging Ca2+ pools were estimated in frog sartorius muscles. A new technique using Sr2+ to extract the rapidly exchanging pool was used. The method avoids problems of kinetic analysis. The results showed that stimulation causes Ca2+ to be translocated from a compartment which exchanges with a time constant of 800 min to a compartment that can be washed out in 15 min. This is likely a transfer from the terminal cisternae to the transverse tubule. Calculations show that this would represent 0.9% of the Ca2+ released in each twitch. After 300 twitches produced by a 1-Hz stimulation, this accumulation could have increased the Ca concentration in the transverse tubules to 70 mM. A marked increase of Ca2+ concentration of this magnitude in the transverse tubules would raise the mechanical threshold for excitation--contraction coupling and would decrease the efficiency of coupling between contraction and excitation. This could be the explanation of the fatigue observed during this kind of stimulation. PMID- 6286072 TI - Calcium currents of frog and insect skeletal muscle fibres measured during voltage clamp. AB - Both vertebrate and invertebrate skeletal muscle fibres have Ca2+ permeability mechanisms which are turned on by depolarization of the surface membrane. In frog muscle, Ca currents are extremely slow and will be scarcely activated during the action potential that normally elicits a twitch. This Ca permeability cannot therefore play any substantial, direct role in excitation--contraction coupling. In insect (Carausius morosus) muscle, Ca currents activate within milliseconds of depolarization, even at low temperature, and may well play at least a triggering role in excitation--contraction coupling. These Ca currents show saturation with increasing [Ca]0, while the instantaneous current--voltage relation rectifies inwards, as expected from a very low [Ca]i. The Ca channel is permeable to Sr2+ and Ba2+. Inactivation of Ca currents under a maintained depolarization depends on Ca2+ carrying inward current, however, rather than on the depolarization itself. PMID- 6286073 TI - Intracellular Ca2+ transients in the cat papillary muscle. PMID- 6286074 TI - Activation heat and latency relaxation in relation to calcium movement in skeletal and cardiac muscle. AB - Measurements of activation heat, initial heat, twitch tension, and latency relaxation were made using thin-layer, vacuum-deposited thermopiles and isometric force transducers, respectively. Experiments were performed on frog skeletal muscle fiber bundles and on rabbit right ventricular papillary muscles at 0, 15, ans 21 degrees C in normal and 1.75X to 2.5X mannitol hyperosmotic bathing solutions. In skeletal muscle, activation heat, obtained by stretching to zero overlap, was only slightly affected by 1.75X hyperosmotic solution and consisted of a fast and a slow component. Both components have a refractory period and a relatively refractory period which can be demonstrated by double pulse stimulation. The twitch potentiators Zn2+ and caffeine increase the total activation heat and the magnitude and rate of the fast component. The temporal relation between the latency relaxation and activation heat is demonstrated. The latency relaxation is independent of the number of sarcomeres in series in a muscle. Activation heat and latency relaxation records from heart muscle are obtained in 2.5X hyperosmotic bathing solution. A model of excitation- contraction coupling is presented which indicates that (1) the downstroke of the latency relaxation monitors the functioning of the Ca2+-permeability or debinding mechanism in the terminal cisternae, (2) the fast component of activation heat monitors the amount of Ca2+ bound to troponin C, and (3) the total amplitude of activation heat is a measure of the total quantity of Ca2+ cycled in a twitch. PMID- 6286075 TI - Ouabain potentiation and Ca release from sarcoplasmic reticulum in cardiac and skeletal muscle cells. AB - In this article, we describe a possible mechanism of ouabain potentiation in heart based on the following findings in cardiac and skeletal muscles of various species. (1) In heart ventricle muscles of frog and guinea pig, the ouabain potentiation is produced without an effect on Ca influx. In both frog and cat heart ventricle muscles, ouabain potentiates the rapid cooling contracture with or without caffeine in a Ca-deprived medium. It follows, therefore, that the ouabain potentiation is produced by an "intracellular" mechanism. (2) In crab single muscle fibers, contractile responses such as twitch, potassium-induced contracture, caffeine-induced contracture, and water-induced contracture are remarkably potentiated if ouabain is present within the fibers by microinjection, whereas the situation is reversed if the drug is given extracellularly. (3) The ouabain potentiated the Ca release from fragmented sarcoplasmic reticulum (FSR) isolated from cat, guinea pig, and frog heart and from skeletal muscles as a result of the procedures used, such as changing the ionic environment. (4) In frog, cat, and guinea pig heart ventricle muscles, a reduction of contractility as a result of pretreatment with urea--Ringer's was completely cancelled by ouabain almost without influencing the membrane depolarization. Based on these findings and others, the deduction was made that the positive inotropic effect of cardiac glycosides on the heart is brought about by potentiation of contraction - Ca release from the intracellular store sites, namely the sarcoplasmic reticulum. PMID- 6286076 TI - Fluorescence and differential light absorption recordings with calcium probes and potential-sensitive dyes in skinned cardiac cells. AB - This report describes an optical system for microspectrophotometry in a single cardiac cell from which the sarcolemma has been removed by microdissection (skinned cardiac cell). This system is attached to the high power inverted microscope used for the microdissection and includes (a) a single variable wavelength microspectrophotometer used to define the spectrum of a given dye or Ca2+ probe; and (b) a dual wavelength, differential microspectrophotometer used to record differentially between the optimum wavelength and a wavelength separated by 25--30 nm. Results are presented using the following optical methods: (a) fluorescence measurements with chlorotetracycline to monitor the amount of Ca2+ bound to the inner face of the sarcoplasmic reticulum (SR) membrane; (b) differential absorption measurements with arsenazo III to measure changes of myoplasmic [Ca2+]free resulting from Ca2+ release from the SR; (c)fluorescence and (or) differential absorption measurements with the potential sensitive dyes merocyanine 540, NK 2367, and di-S-C3(5) to monitor changes of charge distribution on the SR membrane during Ca2+ accumulation in the SR, as well as before and during Ca2+-induced release of Ca2+ from the SR. A small and rapid signal is observed which precedes the Ca2+-induced release of Ca2+ from the SR. It is detected as an increase of CA2+ binding inside the SR with chlorotetracycline and as a "hyperpolarization" with potential-sensitive dyes, while no transient change of myoplasmic [Ca2+]free is detected with arsenazo III. This small and rapid signal preceding the Ca2+ release may be a first hint to an understanding of the mechanism whereby a small increase of [Ca2+]free outside the SR triggers Ca2+ release from the SR. PMID- 6286077 TI - The role of superficially bound calcium in the regulation of myocardial contractility and the positive inotropic response to ouabain. AB - Positive inotropic effects induced in hearts by paired pacing and ouabain treatment were found to be dependent on the content of calcium in a superficial pool, Ca1. Two possible mechanisms for the expansion of this pool were investigated in isolated cardiac myocytes in which the cell surface structures were preserved from proteolytic digestion. Guinea pig ventricles were disrupted mechanically to obtain intact myocardial cells. The cell yield from this preparation was enriched by incubation with Cytodex beads and the characteristics of calcium binding to the cell-bed suspension were determined by continuous flow equilibrium dialysis. Scatchard plots revealed two components for calcium binding; a high affinity (Ka approximately 65 mM-1) and a low affinity (Ka approximately 140 M-1) pool. Ouabain (10(-9) to 10(-6)M) increased, in a dose related manner, the KA for calcium binding to the low affinity sites without affecting the total number of binding sites. The calculated amount of calcium bound to the low affinity pool at [Ca]e = 1.8 mM was closely correlated with the positive inotropic effect (PIE) measured in guinea pig hearts perfused with the same calcium concentration. At [Ca]e = 0.45, 0.9, or 3.6 mM, the response to ouabain (1 X 10(-7) and 5 X 10(-7)M) was greater than expected on the basis of the increase in calcium binding to the low affinity sites. The results suggest that at least one event in the inotropic response to ouabain involves increased affinity for calcium binding to low affinity sites on the cell surface, but that the increase in calcium bound at any [Ca]e is insufficient to produce the PIE directly. PMID- 6286078 TI - Relationship between plasma corticosterone and adrenal epinephrine after diazepam treatment in rats. AB - Following recent observations that diazepam treatment increases adrenal epinephrine in rats, we were interested in studying the possible mechanisms of this action of diazepam on rat adrenal glands. All diazepam treatments studied (1 25 mg . kg-1.day-1 for 10 days) led to an increase in adrenal epinephrine following a linear dose-effect relationship. Since epinephrine synthesis is under neuronal and humoral controls, we investigated their respective importance in the effect of diazepam on the adrenal gland. The denervation of the adrenal gland did not prevent the increase in adrenal epinephrine by diazepam treatment. On the hand, diazepam treatment was shown to cause an increase in plasma corticosterone in parallel with an increase in adrenal epinephrine. Administration of dexamethasone (a synthetic corticoid) and hypophysectomy prevented the increase in adrenal epinephrine and plasma corticosterone resulting from diazepam treatment. We thus conclude that the increase in adrenal epinephrine seen after diazepam treatment is parallel to the increase in plasma corticosterone. Moreover, since the action of diazepam on adrenal epinephrine is prevented by dexamethasone or hypophysectomy, we hypothesize that diazepam is acting on the adrenal cortex via the release of adrenocorticotropic hormone (ACTH). ACTH and corticosterone would be responsible for the increased activity of epinephrine synthesizing enzymes in adrenal medulla. PMID- 6286080 TI - The effect of decomplementation on the infectious course of Sendai virus in mice. AB - The course of intranasal infection of Sendai virus in CBA and DBA mice was investigated in animals decomplemented with purified cobra venom factor. The mice were decomplemented either immediately before inoculation or at 4 days postinfection. Depletion of complement after the infection had been established had no apparent effect on the course of the viral infection in the two strains of mice. In contrast, both strains of mice were protected completely from the lethal effects of an infectious dose of 1 LD50 of virus when the serum C3 levels were depressed by more than 80% during the early stages of infection. The symptoms of morbidity were less pronounced in these animals and there was a delay in the production of hemagglutination-inhibiting antibody. There was no apparent effect on the growth of the virus in lung tissue. The results suggest that the complement system plays a significant pathogenic role during the course of Sendai virus infections in CBA and DBA mice. PMID- 6286081 TI - The somatosensory evoked potential. PMID- 6286079 TI - Central injections of spiperone and GABA: attenuation of angiotensin II stimulated thirst. AB - This study investigated the effects of injecting spiperone or gamma-aminobutyric acid (GABA) into the brain on drinking induced by angiotensin II injected into a lateral cerebral ventricle of the rat. Injections of 12.5 pM of angiotensin II elicited water intakes averaging 10 mL in 15 min with latencies of less than 3 min. Spiperone, a dopamine antagonist injected into the nucleus accumbens produced dose-dependent reductions in water intake and number of laps while increasing the latency to drink. Injections of GABA into the ventral tegmental area produced dose-dependent reductions in water intake and number of laps without altering latency to drink or lap volume. GABA injected into the globus pallidus reduced drinking in a manner similar to that described for spiperone injected into the nucleus accumbens when angiotensin II was the dipsogenic agent but was totally without effect on drinking induced by carbachol injections. These results indicate that a proposed neuronal circuit composed of GABA input to ventral tegmental area neurons, dopamine input from the ventral tegmental area to nucleus accumbens neurons, and subsequent GABA input from the nucleus accumbens to globus pallidus neurons may interact with the goal-directed behaviour, drinking, elicited by central angiotensin II administration. Further, they provide evidence that this neuronal circuit may be part of a functional interface for response initiation. PMID- 6286083 TI - Role of chemotherapy and surgery in the treatment of thoracic metastases from nonseminomatous germ cell testis tumor. PMID- 6286082 TI - Gas gangrene following perforation of the colon. PMID- 6286084 TI - Preoperative chemotherapy for unresectable primary hepatic malignancies in children. AB - Eight children presenting with unresectable primary hepatic malignancies were treated with chemotherapy in an attempt to decrease the size of the tumor. Adriamycin was used in all drug regimens, usually in combination with cyclophosphamide, vincristine, and 5-fluorouracil. Seven children exhibited a pronounced, clinical response with marked reduction in the size of the primary tumor as well as any pulmonary metastases present. Four children were able to have complete, uncomplicated surgical excision of residual disease, and three are alive and well off therapy. One patient with hepatocellular carcinoma had compete disappearance of all disease with chemotherapy alone. An approach utilizing preoperative chemotherapy for extensive hepatic malignancies may permit eventual resection of initially inoperable lesions, with long-term survival for these highly lethal malignancies. PMID- 6286085 TI - Clinical and chemotherapeutic study of hepatocellular carcinoma in Malaysia: a comparison with African and American patients. AB - Twenty Malaysian patients with unresectable primary liver cell cancer were prospectively studied at the General Hospital, Kuala Lampur, and were compared for clinical features with an equal number each of African and American patients being studied by the Eastern Cooperative Oncology Group. The patients received intravenous 5-FU and oral MeCCNU which was used for the first time in an Asian country. Most of the Malaysian patients were Chinese, belonged to younger age groups, and presented with massive hepatomegaly, jaundice, and fever. Toxicity to MeCCNU invariably occurred in the form of leukopenia or thrombocytopenia, but none life threatening. Partial response was seen in 20% of Malaysians as compared to 16% in Americans and none in Africans. Malaysians achieved a median survival of 16 weeks compared to 28 weeks in Americans and only eight weeks in Africans. Malaysian Chinese patients were all HBc Ab + ve. Other factors which may have played an etiologic role in the induction of primary liver cancer included alcohol, Chinese herbal medicines, aflatoxin and habitual use of medicated rubbing oils. PMID- 6286086 TI - The influence of radiation therapy quality control on survival, response and sites of relapse in oat cell carcinoma of the lung: preliminary report of a Southwest Oncology Group study. AB - Two hundred and ninety-eight patients with limited (confined to chest and supraclavicular area, encompassable by a single radiation portal) small cell carcinoma of the lung were entered on Southwest Oncology Group Protocol 7628. Patients were treated with multi-agent chemotherapy and radiation therapy with or without BCG. Radiation therapy quality control analysis, including dosimetric reconstruction and port film review was introduced after the protocol was activated and was retrospectively applied. Patients who were considered major protocol variations had statistically worse survival (40 weeks versus 60 weeks; P = .002), a lesser improvement in response rate after induction chemotherapy (27 versus 48%; P = .05) and a higher chest failure rate (77 versus 55%; P = .047) than evaluable patients. Five patients relapsed in the brain, all associated with chest failure. Quality control is essential in cooperative group studies. PMID- 6286087 TI - Impact of irradiation technique and tumor extent in tumor control and survival of patients with unresectable non-oat cell carcinoma of the lung: report by the Radiation Therapy Oncology Group. PMID- 6286088 TI - Cancer chemotherapy associated symptomatic stomatitis: role of Herpes simplex virus (HSV). AB - The relationship between oral shedding of Herpes simplex virus (HSV) and cytotoxic cancer chemotherapy was studied. HSV seropositive outpatients receiving cytotoxic chemotherapy had HSV recovered from throat washings in eight of 114 patients (7.0%), significantly more often than HSV seropositive outpatients with malignancy who were not receiving cytotoxic chemotherapy (one of 91 patients; 1.1%; P = 0.04). Twenty-eight HSV seropositive chemotherapy patients and 11 HSV seropositive healthy hospital personnel were studied serially 2-3 times per week over a 3-4 week period for oral HSV shedding. Although a comparable percentage of each group shed HSV at least once (57.1% of chemotherapy patients versus 36.4% of controls), chemotherapy patients had a strikingly higher incidence of multiple positive cultures: 40/218 attempts (18.7%) versus 4/156 attempts (2.6%) for controls (P less than 10(-5)). Among chemotherapy patients who developed clinically evident stomatitis, 12 of 14 (85.7%) had HSV recovered compared to four of 14 (28.6%) without lesions (P = 0.004). We conclude that while oral mucosal HSV infection is associated with symptomatic stomatitis following chemotherapy, HSV does not account for all mucosal lesions in chemotherapy patients. PMID- 6286089 TI - Influence of chest radiotherapy in frequency and patterns of chest relapse in disseminated small cell lung carcinoma. A Southwest Oncology Group Study. AB - The value of radiotherapy to the chest (RC) in disseminated small cell lung carcinoma (SCLC) has been questioned. Two protocols for disseminated SCLC from the Southwest Oncology Group (SWOG) have been compared. They were developed four years apart. The first one included radiotherapy (RT), 3000 rad in two weeks, to the primary tumor, mediastinum and supraclavicular areas, while the second one deleted any RC. Multidrug chemotherapy (CT) and brain RT were used in both protocols. Nonresponders to CT were removed from the study. Our main findings are as follows: (1) Initial chest relapses (patients with no initial extrathoracic relapse) have increased from 24-55% when RC is not given (P = 0.0001). Overall chest relapse (adding those patients that relapsed simultaneously in the chest plus other sites) in the second protocol was 73%. (2) Amount of response to CT does not influence the chances for relapse. Even complete responders to CT have a high chance for chest relapse. (3) Sites of relapse without RC are mainly in the primary tumor, ipsilateral hilus and mediastinum. (4) With RC, relapses shift to the chest periphery, mostly to the lung outside the radiotherapy field and to the pleura. (5) The two very different CT regimens have produced similar percentages and duration of response. (6) CT schema with periodic reinductions prolongs duration of response and survival over schema with continuous maintenance. Hence, interruption of CT to allow RC does not seem to adversely influence CT efficacy. From our results and the review of the literature we conclude that: (1) patients with disseminated SCLC that respond to CT should be given RC to decrease chest relapses. (2) A dose of 3000 rad in two weeks seems to be enough to produce a low percentage of chest relapse in disseminated SCLC, as survival of these patients is short and many will die prior to developing chest relapse. However, according to the literature, 4000-4800 rad is probably a more effective dose. (3) More studies and guidelines are needed to outline proper boundaries of radiotherapy fields, to decrease chances of peripheral chest relapses. (4) Median survival might not be a good parameter to evaluate the impact of RC in disseminated SCLC. The study of long-term survivors seems to be more important. PMID- 6286090 TI - Occurrence of anti-retinal ganglion cell antibodies in patients with small cell carcinoma of the lung. AB - This report describes a patient with small-cell carcinoma of the lung associated with blindness. The serum of this patient was tested for immunoreactivity with retinal sections because there was no evidence of tumor metastasis to the central nervous system and because neuroendocrine cells of the lung share common antigens with retinal neurons. The immunoglobulins in the sera from this patient, from two other patients with small-cell carcinoma, from two patients with multiple sclerosis and from three controls were reacted with 8 micrometers thick sections of retina. Dog, cat and human retinal sections were used. Antihuman immunoglobulins that were conjugated to horseradish peroxidase were used as the second antibody to identify the cells which bound the patients' immunoglobulins. A high titer (1:500) antibody level against retinal ganglion cells was found in the patient with small-cell carcinoma and blindness; the antibody reaction was similar with retina from the three species. The large ganglion cells bound the immunoglobulins of the patients with small-cell carcinoma and blindness while the immunoglobulins from the control and multiple sclerosis subjects did not bind these cells selectively at high dilutions. It remains to be shown whether these antibodies have an etiologic significance in the development of blindness. PMID- 6286091 TI - Duct carcinoma in situ. Relationship of extent of noninvasive disease to the frequency of occult invasion, multicentricity, lymph node metastases, and short term treatment failures. AB - Fifty-three breasts resected for a biopsy diagnosis of duct carcinoma in situ were studied with a serial subgross and correlated radiographic method of examination designed to permit quantitation of the extent of the noninvasive lesion in the breast. Overall frequencies of occult invasion and multicentricity were 21 and 32%, respectively. Among 24 lesions 25 mm or larger in extent (average, 63; median, 56 mm) 11 showed occult foci of invasion, 13 had multicentric foci and six had nipple involvement. Among 29 lesions less than 25 mm in extent (average, 10; median, 8 mm) there were no instances of occult invasion, four were multicentric and two had nipple involvement (P = less than 0.05 for multicentricity and occult invasion). Twenty patients with lesions averaging 8 mm in extent are being followed after excision only in an experimental program. There have been three local recurrences at an average follow-up of 44 months. All recurrences occurred ipsilaterally, two were within the prior biopsy site. All patients with recurrence are free of disease following local resection in two and modified radical mastectomy in one. For lesions with associated microcalcifications, the distribution of the mammographic microcalcifications closely approximates the extent of disease as confirmed histologically. These findings suggest that an important predictive factor for the presence of occult invasion and multicentricity in the resected breast is the extent of the noninvasive lesion. PMID- 6286092 TI - Histopathologic classification of small cell carcinoma of the lung: comments based on an interobserver examination. AB - In order to evaluate the consistency in diagnosing the morphologic subtypes of small cell carcinoma of the lung (SCCL), 93 microscopic slides were blindly classified by three panelists according to the World Health Organization's (WHO) Lung Tumor Classifications of 1967 and 1981. Unanimity in the diagnosis of SCCL as the main cell type was obtained in 91 and 94% respectively. With regard to the diagnosis of the various morphologic subtypes unanimity among the three panelists was achieved in 38% according to the WHO 1967 classification and in 54% using the 1981 classification. It is concluded that the criteria used for SCCL as a distinct histopathologic entity were reproducible among the different panelists. However, different criteria were applied in the diagnosis of the morphologic subtypes of SCCL. This fact could explain the contradictory results of clinical studies reported in the literature concerning subtyping of SCCL. PMID- 6286093 TI - The effect of cell size on the pathologic diagnosis of small and large cell carcinomas of the lung. PMID- 6286094 TI - Thorotrast-associated cholangiocarcinoma: an epidemiologic and clinicopathologic study. AB - Six cases of Thorotrast-associated cholangiocarcinoma (TACC) are studied and compared with eight previously reported cases as well as with cases of non-TACC. TACC shares some epidemiologic features both with non-TACC as well as Thorotrast associated angiosarcoma (TAAS) but appears to occur in younger patients than non TACC. There is an apparent increased prevalence of both benign and malignant thyroid disease in the current group of patients with TACC; previous epidemiologic studies have not identified this phenomenon. Thorotrast-associated hepatic fibrosis was present in all 14 TACC cases; otherwise, both non-TACC and TACC are clinically and morphologically similar. Bile duct dysplasia and/or transition to adenocarcinoma, which occurred in all the AFIP cases, are seldom described in case reports of either TACC or non-TACC; in the current study they served to validate the intrahepatic origin of the tumors. PMID- 6286095 TI - Appearance of small cell anaplastic carcinoma of the lung in two patients apparently cured of squamous cell carcinoma of the lung. PMID- 6286096 TI - The significance of the internal mammary lymph nodes in medially located breast cancer. AB - The efficacy of adjuvant radiotherapy for medial breast cancers in Stage I or II disease was studied with a retrospective clinical analysis. All cases of breast cancer free of axillary node metastases at the time of standard or modified radical mastectomy were included. These were grouped by location and size of the primary tumor. There were 76 medial lesions, including those in the 6:00 or 12:00 axis. Seventeen of these had received postoperative radiation. Lateral tumors included 59. The size was based on the largest linear dimension and grouped less than or equal to 2 cm or greater than 2 cm. At the time of initial recurrence, all areas of metastases were determined. The data suggests that: (1) medial breast cancers have a greater rate of recurrence than lateral ones; (2) adjuvant radiation for medial lesions decreases that recurrence rate to the range of lateral tumors; (3) large lesions may have a greater chance for local recurrence as well as medial lesions of all sizes; and (4) the potential problems with local recurrence may be diminished with adequate radiotherapy as an adjunctive measure. When recurrence does occur, prior radiotherapy seems to delay the appearance of that recurrence. PMID- 6286097 TI - Clonal origin of the Philadelphia translocation in chronic myeloid leukemia demonstrated in somatic cell hybrids using an adenylate kinase-1 polymorphism. AB - Hybrid cell lines were derived from fusion of rodent cells with leukocytes from a t(9q+; 22q-)-positive chronic myeloid leukemia (CML) patient carrying a chromosome No. 9-linked adenylate kinase-1 (AK1) polymorphism (AK1 1-2). The AK1*2 allele was consistently expressed when 9q+ was present, whereas the AK1*1 coded isozyme was formed when the normal chromosome No. 9 was present. These results provide additional data confirming the clonal origin of the Ph1 translocation in CML. PMID- 6286098 TI - A nonrandom chromosomal abnormality, del 3p(14-23), in human small cell lung cancer (SCLC). AB - In order to determine whether or not there are specific chromosomal changes in small cell lung cancer (SCLC), karyotypic analyses of 16 continuous SCLC tissue culture lines, three fresh tumor specimens (bone marrow), one direct preparation of bone marrow involved with SCLC, and two lymphoblastoid lines derived from SCLC patients were studied. Cell lines were derived from primary tumor, or metastases to bone marrow, subcutaneous nodules, or pleural fluid; all 16 lines had biochemical and histologic properties characteristic of SCLC. Of the 15 males and 3 females, 6 patients had no prior treatment. All of the 16 cell lines, the 3 fresh specimens, and the direct bone marrow preparation had a common deletion of the short arm of chromosome #3. Use of the shortest region of overlap analysis showed the common deletion was of the short arm in the regions p(14-23). This specific chromosomal abnormality, del 3p, was not found in five non-SCLC cell lines studied and is of major potential biological and diagnostic importance. PMID- 6286099 TI - Burkitt's lymphoma with 2/8 translocation: a case report with special reference to the clinical features. AB - A case of Japanese Burkitt's lymphoma (BL) with a t(2; 8) (p11; q24), a variant translocation in BL, is reported. The patient was a 45-year-old woman in whom a subcutaneous right axillary tumor first occurred. Remission was not achieved despite extensive chemotherapy. Of the four nonendemic BL, two endemic BL, and one nonendemic Burkitt-type acute lymphocytic leukemia (ALL-L3) cases with a t(2; 8) reported so far, including the present case, four (two nonendemic and two endemic) were positive for Epstein-Barr virus-determined nuclear antigen (EBNA) and two revealed extremely high antibody titers against Epstein-Barr virus (EBV), the remaining one not having been tested for EBV. Thus, a possible close relationship between the t(2; 8) and EBV infection has to be considered. The t(2; 8) in nonendemic BL seems to occur more often in adults than in children. PMID- 6286101 TI - Priming of human myeloid leukemic cell lines HL-60 and U-937 with retinoic acid for differentiation effects of cyclic adenosine 3':5'-monophosphate-inducing agents and a T-lymphocyte-derived differentiation factor. AB - The human cell lines HL-60 (acute promyelocytic leukemia) and U-937 (histiocytic monoblast-like lymphoma) differentiate to functionally mature cells by incubation with retinoic acid (RA). This differentiation is potentiated by agents known to increase intracellular cyclic adenosine 3':5'-monophosphate (cAMP) levels. The present study shows that these cells can be primed for differentiation by treatment for approximately one day with RA followed by exposure to a cAMP inducing agent. The reverse sequence was ineffective. Thus, HL-60 could be primed by incubation for less than 20 hr with 10 nM RA to respond by differentiation to the addition of 10 nM prostaglandin E2 or 1 nM cholera toxin, whereas 10 nM RA alone was almost inactive. RA-primed HL-60 also responded with differentiation to a concentration of T-lymphocyte-derived differentiation-inducing factor which alone was inactive. U-937 primed by incubation for 24 hr with 100 nM RA responded to cAMP-inducing agents and differentiation-inducing factor, which alone were inactive on this cell line. Priming of these cell lines does not depend on the normal rate of protein synthesis, as it occurs even better in the presence of a concentration of cycloheximide that inhibits growth completely, suggesting that a decrease in synthesis of some protein(s) favors RA-induced differentiation. Cycloheximide alone produced some priming of HL-60 but not of U-937. HL-60, but not U-937, primed with RA responded to adenosine triphosphate and other nucleoside triphosphates, consistent with the notion that modulation of the RA effect may be mediated through protein kinase activity at the plasma membrane. The finding that myeloid cell lines, like HL-60 and U-937, blocked at a late stage of maturation can be primed by RA to respond to cAMP-inducing agents and differentiation-inducing factor may improve our understanding of the arrest in maturation typical of some forms of leukemia. PMID- 6286100 TI - Induction of differentiation of the human histiocytic lymphoma cell line U-937 by retinoic acid and cyclic adenosine 3':5'-monophosphate-inducing agents. AB - The monoblast-like human histiocytic lymphoma cell line, U-937, is induced to differentiate into monocyte-like cells by incubation with 0.1 to 1.0 microM retinoic acid (RA). These induced cells are phagocytic, reduce nitroblue tetrazolium, and show an increased hexose monophosphate shunt activity, consistent with monocyte-like cells. Prostaglandin E, cholera toxin, or dibutyryl cyclic adenosine 3':5'-monophosphate, all inactive alone, increased markedly the extent of RA-induced differentiation of U-937. These data suggest that the intracellular level of cyclic adenosine 3':5'-monophosphate is of importance for expression of the RA-induced effect. Responsiveness to RA seems to be limited to those leukemic myeloid cells blocked at a relatively late stage of maturation, like the promyelocytic HL-60 and the monoblast-like U-937 cell lines. PMID- 6286102 TI - Neutralization of feline leukemia virus with feline antisera to leukocyte alloantigens. AB - The possibility that normal cellular antigens might serve as targets for antibody neutralization of the feline leukemia virus was investigated. Xenospecific antiserum directed to normal feline cells was shown to inactivate feline leukemia virus grown in fibroblasts. Cat antisera to normal feline leukocyte alloantigens were then prepared, after which persistent viremia was induced in the donor cats. Such alloantisera neutralized virus taken from plasma of the appropriate cat but did not neutralize virus from a different cat. The virus neutralization was dependent on the presence of complement. These results indicate that alloantigens are present at the virus surface and raise the possibility that such antigens may play a role in the natural immune response directed to retrovirus infections. PMID- 6286104 TI - Benzo(a)pyrene metabolism by rat liver microsomes: effects of adding purified glutathione S-transferases A, B, and C. AB - We have examined the effects of adding glutathione and isolated cytosolic glutathione S-transferases A, B, and C to rat liver microsomes metabolizing benzo(a)pyrene. Addition of glutathione alone resulted in the conjugation of 15 to 20% of the total metabolites of benzo(a)pyrene, and this conjugation could be inhibited almost entirely by bromosulfophthalein (an inhibitor of glutathione S transferases), indicating that it is catalyzed by the glutathione S-transferase present in microsomes. Addition of purified cytosolic glutathione S-transferases A, B, and C yielded about 30 to 40% conjugate formation. Analysis of metabolites by high-pressure liquid chromatography demonstrated that the formation of 4,5 diol of benzo(a)pyrene was decreased by at least 80% by conjugation and that the 7,8-diol was also decreased significantly (40 to 60%). In addition, it was found that glutathione S-transferase B is capable of conjugating benzo(a)pyrene 1,6- and 3,6-quinones. PMID- 6286103 TI - Temporal dynamics of cortisol and dexamethasone prevention of benzo(a)pyrene induced morphological transformation of Syrian hamster cells. AB - The temporal aspects of the anticarcinogenic action of cortisol were investigated by measurement of the frequency of carcinogen-induced morphological transformation of Syrian hamster embryo cells relative to treatment regimen. Treatment of hamster cells with 10(-5) to 10(-11) M cortisol, dexamethasone, corticosterone, 17 beta-estradiol, progesterone, or testosterone did not result in morphological transformation. However, cortisol or dexamethasone treatment beginning 48 hr before the carcinogen caused a dose-dependent reduction in the transformation frequency associated with benzo(a)pyrene [B(a)P] or ultraviolet irradiation treatment. The frequency of B(a)P transformation was reduced 54, 66, and 84% by 10(-11), 10(-10), and 10(-9) M cortisol, respectively, without altering colony formation efficiency. Dexamethasone was equipotent as cortisol as an inhibitor of B(s)P, N-methyl-N'-nitro-N-nitrosoguanidine, or ultraviolet irradiation-induced transformation. Sex steroids, non-steroid antiinflammatory compounds, and steroids without antiinflammatory activity also reduced the transformation frequency but only at concentrations that reduced colony formation. Cortisol inhibition of transformation increased as steroid exposure occurred closer (48, 24, and 8 hr) yet prior to B(a)P treatment. In contrast, if cortisol was added simultaneously with or up to 72 hr after B(a)P, the frequency of transformation was unaffected. Dexamethasone addition after B(a)P, however, reduced the transformation frequency. Thus, physiological concentrations of cortisol when present prior to carcinogen exposure can prevent carcinogen-induced morphological transformation. The ability of cortisol to inhibit ultraviolet irradiation-induced transformation indicates that the anticarcinogenic activity may be mediated by steroid-induced cellular changes which are independent to metabolic activation of the carcinogen. PMID- 6286105 TI - Correlations of inheritance and expression between a tumor gene and the cellular homolog of the Rous sarcoma virus-transforming gene in Xiphophorus. AB - Neoplastic transformation of pigment cells in the teleostean fish Xiphophorus is mediated by a cellular oncogene (Tu). Normally. Tu is suppressed by multiple regulating genes (R). Depending on impairment and loss of R genes, Tu is permitted to express itself phenotypically. In the pigment cell system, different degrees of Tu expression lead to small spots of transformed cells or to benign or malignant melanoma. All neoplastic and nonneoplastic cells of all Xiphophorus genotypes tested thus far appear to contain the cellular homolog (c-src) of the avian sarcoma virus oncogene (v-src). The evidence for this stems from the detectability of a Mr 60,000 phosphoprotein with associated kinase activity (pp60c-src) that reacts with antiserum against viral pp60src. We followed the inheritance of Tu (identified by spots and melanomas) compared to the expression of c-src identified by the pp60c-src-associated protein kinase). By quantitative determination of kinase activity in immunoprecipitated pp60c-src from fish showing different degrees of Tu expression, we have investigated whether there exists a correlation between the expression of c-src and Tu. In genotypes with the same genetic background, cells from Tu-containing fish express more pp60c-src than do cells from fish lacking Tu. In genotypes carrying a Tu gene and which show differences in the amount of gene expression due to a different extent of repression by regulating genes, analysis of kinase activity revealed that an increase of Tu expression is correlated with an elevated level of pp60c-src associated kinase activity. Our findings may indicate that c-src activity in Xiphophorus is modulated by the Tu gene product or that Tu and c-src are regulated coordinately. PMID- 6286106 TI - Autologous bone marrow transplantation in the therapy of small cell carcinoma of the lung. AB - Ten patients with small cell carcinoma of the lung were entered into a chemotherapeutic treatment program consisting of cyclophosphamide, vincristine, Adriamycin, and 1-(2-chloroethyl)-3-cyclohexyl-1-nitrosourea. Two courses of combination chemotherapy were administered to each patient followed by a third course with the same doses of drugs used on Course 2 but with autologous bone marrow transplantation given 24 to 48 hr after drug infusion. No differences could be detected between Courses 2 and 3 in terms of the magnitude, timing, or degree of myelosuppression. Serial bone marrow biopsies documented a progressive decline in granulocyte-macrophage colony-forming units in culture per mg bone marrow medullary core from 138 +/- 179 (S.D.) prior to chemotherapy to 7 +/- 11 after the marrow transplant recovery (p = 0.05). These data suggest that autologous bone marrow transplantation does not reduce the myelosuppression seen following the drugs used in this study at the dosages used. Autologous bone marrow transplantation may be useful only in the setting of marrow lethal therapy. Its usefulness in shortening recovery time from nonlethal therapy appears questionable. PMID- 6286108 TI - Specific high-affinity binding of the phorbol ester tumor promoter 12-O tetradecanoylphorbol-13-acetate to isolated nuclei and nuclear macromolecules in mouse epidermis. AB - The tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA) binds reversibly and with high affinity and specificity to nuclear macromolecules in mouse epidermis. The dissociation constants determined from Scatchard analysis of epidermal nuclei and nuclear macromolecules are 3.58 +/- 0.66 (S.E.) and 2.18 +/- 0.54 nM, respectively. The solubilization of TPA receptors from epidermal nuclei by DNase I was examined. Following a 20-min digestion at 22 degrees, more than a 2-fold increase in specific TPA binding was observed in the supernatant relative to non-nuclease-treated nuclei (0.71 versus 0.32 pmol/mg protein, respectively). Our data indicate that epidermal nuclei contain saturable and specific TPA binding macromolecules and that these binding components may be associated with regions of chromatin that are preferentially susceptible to nucleolytic cleavage. These data suggest the existence of nuclear receptors for the phorbol ester tumor promoters. These observations may necessitate a more critical assessment of plasma membrane binding as the sole binding site responsible for triggering the multistep process of tumor promotion in mouse epidermis. PMID- 6286107 TI - Role of estrogen and prolactin in the growth and receptor levels of N nitrosomethylurea-induced rat mammary tumors. AB - Forty-eight of 81 (59%) of N-nitrosomethylurea-induced rat mammary tumors regressed in average to almost one-half of the original size 10 days after ovariectomy (ovax) (hormone responsive), while 33 remained essentially unchanged (hormone resistant). AT 20 days after ovax, further decline in hormone-responsive tumors was observed when the rats were treated daily with 0.9% NaCl solution on the tenth day after ovax. Treatment for the same length of time with estrogen either alone or in combination with bromocryptine (to effectively suppress serum prolactin level) prevented tumor regression in hormone-responsive tumors. A similar effect was observed when rats were treated with perphenazine (to stimulate endogenous prolactin secretion) either alone or in combination with the antiestrogen tamoxifen. Estrogen receptors (ERs) significantly declined after ovax. Treatment with estrogen or perphenazine did not have any significant effect on ER level. Progesterone receptors (PGRs) became virtually undetectable after ovax. Treatments with estrogen, estrogen plus bromocryptine, and perphenazine plus tamoxifen but not perphenazine alone were able to partially restore PGRs although this effect was of borderline statistical significance. ER and PGR levels were not significantly different between hormone-responsive and -resistant tumors within each group. We conclude that both estrogen and prolactin play a role in the growth of the N-nitrosomethylurea-induced rat mammary tumor. Changes in ER and PGR levels did not correlate with tumor growth under the present experimental conditions. PMID- 6286109 TI - Differential effects of retinoic acid and 7,8-benzoflavone on the induction of mouse skin tumors by the complete carcinogenesis process and by the initiation promotion regimen. PMID- 6286110 TI - Actions of cis-diamminedichloroplatinum on cell surface nucleic acids in cancer cells as determined by cell electrophoresis techniques. PMID- 6286111 TI - Induction of ornithine decarboxylase in hamster tracheal epithelial cells exposed to asbestos and 12-O-tetradecanoylphorbol-13-acetate. PMID- 6286112 TI - Effects of cyclophosphamide and polycyclic aromatic hydrocarbons on cell growth and mixed-function oxidase activity in a human colon tumor cell line. AB - Human colon tumor cells (cell line LS174T) retain a cytochrome P-450-containing drug metabolism system capable of hydroxylating polycyclic hydrocarbons and the anticancer drug cyclophosphamide. The hydroxylation of benzo(a)pyrene by human colon tumor cells is highly inducible. Phenobarbital plus hydrocortisone induce benzo(a)pyrene hydroxylation activities 10-fold, while benz(a)anthracene induces the rate of hydroxylation 30-fold. Cytochrome P-450 specific content is increased 2- to 3-fold by treatment with phenobarbital plus hydrocortisone and benz(a)anthracene, respectively. Addition of cyclophosphamide alone results in no increase in hydroxylation activities but causes a decrease in cell growth rate. The combination of cyclophosphamide with either of the inducers phenobarbital plus hydrocortisone or benz(a)anthracene results in markedly enhanced inhibition of cell growth as judged both by a decrease in the number of cells per plate and in the incorporation of [3H]thymidine into DNA. Thus, these data show that cyclophosphamide is cytotoxic to human colon tumor cells and that the cytotoxicity is enhanced by simultaneous administration of benz(a)anthracene or phenobarbital plus hydrocortisone to the tissue cultures. PMID- 6286113 TI - Expression of type C viral glycoproteins on P815 cells: higher expression of Mr 70,000 glycoprotein-containing glycoprotein on immunogenic variants. AB - Cells from P815 mastocytoma, a methylcholanthrene-induced tumor, were investigated for the presence of type C viral proteins on their surface. Little or no precipitation of purified Rauscher murine leukemia virus Mr 70,000 glycoprotein was observed with sera of mice immunized with P815 cells. However, a significant precipitation was obtained with sera from syngeneic or allogeneic mice immunized with P815 tumor cell variants obtained by mutagenesis. This difference was not detected when the sera were tested for precipitation of a pure Rauscher murine leukemia virus Mr 30,000 protein. The positive sera also precipitated a Mr 90,000 glycoprotein present on the membrane of all P815 cells. This glycoprotein was shown to contain endogenous ecotropic murine leukemia virus Mr 70,000 glycoprotein. On the membrane, a separate glycoprotein with a molecular weight of 90,000 containing gag gene products was also detected with a panel of reference antibodies. The level of glycoprotein containing Mr 70,000 glycoprotein on the cell membrane of the mutagenized cell variants was higher compared to the original P815 cells. The level of expression of the glycosylated gag precursor was the same on the original P815 cells and on the mutagenized cell variants. PMID- 6286114 TI - Tumoricidal effect of macrophages exposed to adriamycin in vivo or in vitro. AB - Peritoneal macrophages from BD IX rats collected 24 hr after an i.p. injection of ADriamycin (10 mg/kg) were cytotoxic to syngeneic cancer cells in culture. In contrast, incubation in vitro in Adriamycin solutions did not evoke tumoricidal activity in peritoneal macrophages, whatever the incubation time (from 1 to 24 hr) and the Adriamycin concentration (from 1 ng to 100 micrograms/ml). Macrophages incubated with Adriamycin in vitro accumulated the drug in their nuclei, whereas macrophages from animals receiving Adriamycin in vivo accumulated it is cytoplasmic vacuoles. Early observation of peritoneal cells after in vivo exposure to Adriamycin shows that Adriamycin is concentrated in mast cell granules which are released and then phagocytosed by peritoneal macrophages. Mast cells exposed to Adriamycin in vitro can induce macrophages to become cytotoxic. These facts explain the difference between macrophages exposed to Adriamycin in vivo and in vitro. Adriamycin fluorescence appears in nuclei of cancer cells incubated with in vivo-labeled macrophages, suggesting that macrophages can directly transfer the drug into cancer cells and therefore play a role in the Adriamycin antitumor effect. PMID- 6286115 TI - Growth of human hepatoma cells lines with differentiated functions in chemically defined medium. AB - A human hepatoma cell line, HuH-7, which was established from a hepatocellular carcinoma, was found to replicate continuously in a chemically defined medium when the medium was supplemented with Na2SeO3. The cells grew better in this medium than in serum-containing medium without any adaptation period. Other established human hepatoma and hepatoblastoma cell lines, HuH-6 cl-5, PLC/PRF/5, huH-1, and huH-4, also grew in the defined medium. Although HLEC-1 cells failed to proliferate continuously with Na2SeO3 alone, they grew if a cell-free conditioned medium from HuH-7 cells was added to the medium. These cell lines, except the HLEC-1 cell line, produced the following human plasma proteins among those examined: albumin, prealbumin, alpha 1-antitrypsin, ceruloplasmin, fibrinogen, fibronectin, haptoglobin, hemopexin, beta-lipoprotein, alpha 2 macroglobulin, beta 2-microglobulin, transferrin, lipoprotein, alpha 2 macroglobulin, beta 2-microglobulin, transferrin, Complement Components 3 and 4, and alpha 1-fetoprotein. Beside plasma proteins, the media from HuH-7, HuH-6 cl 5, PLC/PRF/5, and huH-1 contained anti-carcinoembryonic antigen-reactive proteins, and those from PLC/PRF/5, huH-1, and huH-4 medium contained hepatitis B surface antigen. These proteins were detected during periods of serial cultivation over 9 months under the above culture conditions. The hepatoma cell lines grown in the fully defined synthetic medium may provide a new approach for investigating the growth and metabolism of human hepatoma cells in vitro. PMID- 6286116 TI - Stereotactic interstitial irradiation for the treatment of brain tumors. AB - Eighteen patients harboring untreated, inaccessible, slow-growing tumors or recurrent malignant primary or metastatic brain tumors were treated by the stereotactic implantation of permanent or removable radioactive seeds (192Ir, 198Au, 125I). Seed strength was selected to deliver 10,000-12,000 rad to the tumor's periphery for 192Ir, 9000-12,000 rad for 125I, and 4000-7500 rad for 198Au. Three of six patients implanted with 192Ir seeds had objective responses for 2, 8, and 12 months, and a fourth stabilized for 20 months. Five of 9 patients treated with 198Au were evaluable, three of whom responded for 3, 5, and 20 months. Two patients who received 125I have been evaluated to date and are early failures. Two patients who received 125I have been evaluated to date and are early failures. Because of its higher dose rate 198Au is superior for the faster-growing malignant tumors; but because 125I is easier to shield, 125I seeds with greater activity are being developed for evaluation. PMID- 6286117 TI - How heterogeneous are gliomas? PMID- 6286118 TI - Parenteral intravenous nutrition (PIVN) as an adjunct to chemotherapy in small cell anaplastic lung carcinoma. AB - A randomized trial was initiated to compare the effects of PIVN-associated chemotherapy (adriamycin, vincristine, VP16-213 and cyclophosphamide) versus as chemotherapy control group in patients with small-cell lung neoplasias. The results obtained are preliminary. The test group included ten patients whereas nine were followed in the control group. PIVN was scheduled each day the patient underwent chemotherapy. Each patient received 1550 kcal/day, which included 10% glucose, 20% lipids, and amino acids which may or may not be mixed in the same infusion bottle. The results show three PIVN patients presently in complete remission at the end of three courses of treatment compared to two over the same time period in the chemotherapy control group. Of all patients who can be evaluated at this time, five out of six PIVN patients are in complete remission compared to four out of five in the control group. There was no significant difference in either general health or side effects, such as nausea and vomiting. The median length of time for which a low white cell or platelet count was noted was the same for both groups. PMID- 6286119 TI - Upper hemibody and local chest irradiation as consolidation following response to high-dose induction chemotherapy for small cell bronchogenic carcinoma--a pilot study. AB - Encouraging results of the combination of upper hemibody irradiation (UHBI) and local chest irradiation (LCI) combined withh standard-dose chemotherapy in patients with extensive small cell bronchogenic carcinoma led us to a second pilot study utilizing the same radiation program combined wit high-dose induction chemotherapy. Fourteen patients with small cell bronchogenic carcinoma, five with extensive disease and nine with localized disease, were treated with cyclophosphamide (1.5 g/m2 iv, Days 1 and 22), lomustine (70 mg/m2 orally, Day 1), and methotrexate (15 mg/m2 twice weekly during Weeks 2, 3, 5, and 6). UHBI (600 rads) was given during Week 6 in a single dose and LCI was given during Week 7 (2000 rads/five fractions) to the tumor and mediastinum. Maintenance chemotherapy began in Week 12 with cyclophosphamide (700 mg/m2 iv every 3 weeks) and lomustine (70 mg/m2 orally every 6 weeks). Twelve patients were evaluable for response and toxicity (eight with limited disease). There were three complete response and seven partial responses after induction chemotherapy. After completion of the consolidation radiation therapy, all 12 patients had a response: six complete responses and six partial responses. Acute toxic effects included nausea and vomiting in eight patients, fever in five, and hypotension and angina in one. Subacute toxic effects included nausea, vomiting, and dehydration in two patients who required hospitalization, prolonged aplasia in one, reversible radiation esophagitis in three. Three patients had radiation pneumonitis including one with bilateral diffuse disease that led to death from respiratory failure. Only two of 12 patients received their maintenance therapy on schedule. Treatment failures occurred within the LCI field in seven patients and in distant metastatic sites in six. The median time to first relapse was 7 months and the median survival was 9 months. Because of toxicity, treatment delays, and poor survival in this group of patients, we cannot recommend this combined modality approach. PMID- 6286121 TI - Platinum concentrations in human glioblastoma multiforme following the use of cisplatin. AB - A 24-year-old female with glioblastoma multiforme was not given a nitrosourea because of interstitial lung disease. Cisplatin was administered monthly for five cycles until disease progression. A second debulking procedure was performed and tumor tissue assayed for the presence of platinum. Levels were measurable 7 weeks after the course of treatment. PMID- 6286120 TI - Phase II trial of m-AMSA in hepatocellular carcinoma: a Southwest Oncology Group Study. AB - Twenty-three patients with heptocellular carcinoma were treated with m-AMSA at a dose of 120 mg/m2 iv repeated at 4-week intervals. Toxicity was primarily hematologic. Partial responses occurred in two of 14 previously treated patients and in one of nine previously untreated patients. The overall activity of m-AMSA in patients with hepatocellular carcinoma appears minimal. PMID- 6286122 TI - Phase II study on DON in patients with previously treated advanced lung cancer. PMID- 6286123 TI - Leukemia occurring after treatment of small cell lung cancer. PMID- 6286125 TI - Characterization of the dopamine stimulated adenylate cyclase in the pedal ganglia of Mytilus edulis: interactions with etorphine, beta-endorphin, DALA, and methionine enkephalin. AB - The dopamine-stimulated adenylate cyclase activity was studied both in vivo and in vitro in the central nervous system of the bivalve mollusc Mytilus edulis. Dopamine, epinine, and apomorphine stimulated the enzyme system. Fluphenazine, haloperidol, chlorpromaxine, and to a lesser extent BOL inhibited the dopamine stimulated adenylate cyclase. Etorphine, beta-endorphine, DALA, and methionine enkephalin depressed cyclic AMP levels. This phenomena was naloxone reversible. In addition, the opioids inhibited the stimulation of adenylate cyclase by dopamine. This phenomena was also naloxone reversible. The study demonstrates an interaction among dopamine, the opioids, and cyclic AMP. PMID- 6286124 TI - Batrachotoxinin-A 20-alpha-benzoate: a new radioactive ligand for voltage sensitive sodium channels. AB - Batrachotoxinin-A 20-alpha-benzoate (BTX-B), an analog of the potent depolarizing agent batrachotoxin (BTX), was prepared by selective esterification of naturally occurring batrachotoxin-A with benzoic acid. BTX-B depolarized rat phrenic nerve diaphragm preparations with a time course and concentration dependence virtually indistinguishable from that of BTX. A specific, saturable component of equilibrium binding of [3H]BTX-B to mouse cerebral cortex homogenates was measured, described by an equilibrium dissociation constant of 0.7 microM and a maximum number of binding sites of 90 pmol per gram of tissue (wet weight). Specific binding is inhibited by BTX and other BTX analogs, veratridine, and grayanotoxin but is unaffected by tetrodotoxin and cevine. Under conditions of this assay, neither crude Leiurus quinquestriatus scorpion venom nor purified sea anemone toxin have any effect of specific binding. The data support the conclusion that BTX-B interacts with a recognition site associated with voltage sensitive sodium channels which is identical to the recognition site for BTX. PMID- 6286126 TI - Ionic mechanism of a voltage-dependent current elicited by cyclic AMP. AB - Intracellular pressure injection of cyclic AMP induces a slow voltage-dependent inward current in some neurons of Aplysia californica. The time course, voltage dependence, and ionic sensitivities of this response are nearly identical to those of the voltage-dependent calcium current induced by serotonin in the same preparation. The response to cyclic AMP is unaffected by changes in the extracellular concentration of chloride or potassium. The current is slowly but minimally reduced by a sodium-free solution. The calcium channel blocker, cadmium, blocks the current elicited by injection of cyclic AMP. The data presented here suggest that cyclic AMP can induce a voltage-dependent calcium current. PMID- 6286128 TI - The effect of S100 protein on the plasma membrane function of neurons. AB - The protein S100 markedly increases the net intake of GABA across the plasma membrane of Deiters' neurons which have GABA receptors on their surfaces. This membrane function of S100 was found by using a new microtechnique. Plasma membranes of such cells have been freshly prepared by freehand microsurgery and are tightly fixed over a 30-micrometers phi hole between two compartments of a microchamber containing 2.0 mM GABA in 7.5 microliters and 0.2 mM GABA in 75 microliters, respectively. The transport of GABA has been determined after incubation of the membrane for from 30 sec to 10 min at 29 degrees C. GABA is transported at a rate of 145 ng in 3 min over a 700-micrometers2 membrane area. S100 in its calcium form reacts with the membrane and increases GABA transport by 20% which is ATP dependent and inhibited by ouabain and ruthenium red. The kinetics of the transport furthermore prove that GABA transport across the plasma membrane is an active process. PMID- 6286127 TI - Aminergic neurons: state control and plasticity in three model systems. AB - Aminergic neurons have particular functions in many systems, and in this review their role is discussed and compared in three systems: those parts of the central nervous system controlling sleep and waking in the cut; the superior cervical ganglion: and the isolated nervous system of Aplysia. In the cat the aminergic neurons are most important in a waking state during which time external information is received, processed, and can be retrieved, and during which time habituation and sensitization occur. Aminergic neurons appear to have similar roles in state control in plasticity in both the Aplysia nervous system and the superior cervical ganglion. The striking similarities in the role of aminergic neurons in these three systems support the speculation that aminergic neurons have uniquely important roles in regulation of the plastic properties of neurons. PMID- 6286129 TI - Immunoreactive ACTH/beta-endorphin neurons in the tubero-infundibular hypothalamus of rats. AB - In the preinfundibular portion of male rat hypothalamus, the arcuate-median eminence region was examined after staining with anti-porcine ACTH 1-39 serum. In several cases, anti-beta-endorphin serum was also employed. Both sera stain the same cell bodies. The cell bodies of the immunoreactive neurons are scattered in the subependymal layer, arcuate nucleus and lateral tuberal region. Fibers originating from these cell bodies are distributed extensively throughout these regions. They make synaptic contacts on immunonegative fibers in the arcuate nuclei, or terminate directly in the perivascular space in the internal layer of the median eminence. The findings suggest that the ACTH/beta-endorphin neuronal system may serve neuronally and humorally as an intrahypothalamic constituent of the hypothalamic pituitary regulation system. PMID- 6286130 TI - Degranulation and appearance of vesicular inclusions in canine C cells after administration of antithyroid drug. AB - When stimulated by the feeding of ethylenethiourea for a period of 6 months, dog thyroid glands increased to approximately 30 times their normal size. Not only follicular cells but also C cells had hyperplastic and hypertrophic features, compared with normal controls there was in the C cells a marked decrease in secretory granules immunoreactive for calcitonin. Furthermore, vesicular inclusions of various sizes and dilated nuclear enveloped, which showed a positive immunoreaction for calcitonin, were observed in the C cells. These findings indicate that the antithyroid drug interferes with synthesis of calcitonin by C cells probably by inhibiting the conversion of a larger precursor to calcitonin by C cells, probably by inhibiting the conversion of a larger precursor to calcitonin. PMID- 6286131 TI - Ca entry at rest and during prolonged depolarization in dialyzed squid axons. AB - Ca influx has been studied in squid axons under internal dialysis control. In axons dialyzed with "normal" physiological conditions (Nai = 40-50 mM, Cai2+ = 0.06-0.1 microM, ATP = 2 mM, Ki = 310 mM), 70% of the resting Ca influx is sensitive to external TTX (K0.5 congruent to 5 nM), 20% of it can be accounted by the reversal of the Na-Ca exchange, and the remaining fraction (10%) is insensitive to TTX, D-600, and Nai. The Ca antagonic drug D-600 (50-100 microM) has an inhibitory effect on the resting Ca influx. This compound was found to affect both the TTX sensitive and the Nai-dependent Ca influx components. In the presence of Nai and ATP, Cai2+ activates the carrier mediated Ca entry (Nai dependent Ca influx). Most of the activation occurs in the submicromolar range of Cai2+ concentrations (K0.5 congruent to 0.6 microM). In the absence of Nai and/or ATP, no activation of Ca influx by Cai2+ was found up to about 5 microM Cai2+. Prolonged depolarization with high Ko causes an increase in Ca influx sustained for long time (minutes). Depolarizing the axons by removing Ki causes the same effect. This depolarization-induced Ca entry was only observed in axons containing Nai. In the absence of Nai, Ca influx decreases with increasing Ko. The activation of the carrier mediated Ca entry (electrogenic Na/Ca exchange) by membrane depolarization was found to be markedly dependent on the magnitude of Ca2+ i. Increasing the magnitude of Ca2+ i from 0.1 to 0.6 microM causes a ten fold increase in the extra Ca influx induced by a K-depolarization. PMID- 6286133 TI - Opioid receptor impairment--underlying mechanism in "pain diseases"? AB - The scarce or absent analgesic effect exhibited by morphine on pain from migraine attack and the poor inhibition of the spasmogenic effect of 5-HT (tested on the hand dorsal vein, computerized venotest) suggest the hypothesis of an opioid receptor deficiency in headache sufferers. Since endogenous opioids control the nociception, the sense of well being, and the vegetative balance, an opioid receptoral hypofunction could be the background of the headache and central panalgia, where the trinity pain, anhedonia, and dysautonomia are the characteristic features. PMID- 6286132 TI - Oxalate transfer across the membranes of sarcoplasmic reticulum during the uptake of Ca++. AB - The kinetic profile of Ca++ uptake in the presence of oxalate is biphasic. An initial phase independent on oxalate is followed by an oxalate-dependent phase delayed in time. The ionophore X-537A only abolishes the net Ca++ uptake if added before the onset of the oxalate phase. However, during this phase, X-537A suddenly releases an amount of Ca++ similar in quantity to that released in the initial phase. The delay of the oxalate-dependent phase is a function of pH. At pH of about 5.5, the oxalate phase and simultaneous calcium oxalate precipitation would theoretically start at the beginning, with no delay. Ejection of protons during Ca++ uptake is strongly depressed by oxalate, but not by other organic anions which do not trap Ca++. It is suggested that oxalate is transferred to the inside of the vesicles as a monoprotonated species at expense of protons ejected by the Ca++-pump during the uptake of Ca++. PMID- 6286134 TI - Biochemical indices of sympathetic activity in migraine. AB - Plasma levels of adrenaline, noradrenaline, cAMP, free fatty acids and serum DBH, were estimated in a group of ten migrainous patients before, during and after a migraine attack. All parameters, apart from adrenaline, showed a statistically significant elevation during the headache period. As the blood concentration of the above substances taken in combination is the most reliable index of sympathetic activity available at present, these findings indicate that there is considerable sympathetic dysfunction during the migraine attack. PMID- 6286135 TI - Molecular genetics of symbiotic nitrogen fixation. PMID- 6286136 TI - Rat kappa-chain J-segment genes: two recent gene duplications separate rat and mouse. AB - We have cloned DNA segments containing the Jk genes from LOUVAIN rat liver, and have determined their nucleotide sequence. Seven readily identifiable Jk-coding regions (six expressible) are evident in the rat, compared with five in the mouse (four expressible). The two additional J segments in the rat appear to be the result of two sequential gene duplications occurring since the divergence of rats and mice. The first involved a homologous but unequal crossing-over in a 14 bp region spanning the 3' end of the coding region of J1 and J2. The second involved a crossing-over following unequal pairing of the two newly duplicated regions. We propose that the probability of a second duplication was greatly increased following the first as a result of the increased target for unequal pairing (370 bp of good homology versus 27 bp in the original pairing). Comparisons of rat and mouse J genes show a surprisingly high degree of sequence conservation, both inside and outside the coding regions, similar to the pattern we reported previously for the kappa constant-region gene. This provides additional evidence that constraints exist on the nucleotide sequences of these genes independent of the function of the encoded proteins. PMID- 6286137 TI - Evidence for supercoiled hepatitis B virus DNA in chimpanzee liver and serum Dane particles: possible implications in persistent HBV infection. AB - In chimpanzee hepatitis B virus (HBV) carriers, the mechanism of viral persistence has been examined by analyzing viral DNA molecules in liver and serum. Chimpanzee liver DNA contained two extrachromosomal HBV DNA molecules migrating on hybridization blots at 4.0 kb and 2.3 kb. There was no evidence for integration of HBV DNA into the host genome. The extrachromosomal molecules were distinct from Dane particle DNA and were converted to linear 3.25 kb full-length double-stranded HBV DNA on digestion with Eco RI. Nucleases S1 and Bal 31 converted "2.3 kb" HBV DNA to 3.25 kb via an intermediate of "4.0 kb" apparent length. The HBV DNA molecule that migrated at 2.3 kb represents a supercoiled form I of the HBV genome, and the molecule that migrated at 4.0 kb represents a full-length "nicked," relaxed circular form II. Evidence for supercoiled HBV DNA in serum Dane particles was obtained by production of form II molecules upon digestion with nuclease S1 or Bal 31. It is proposed that most Dane particles represent interfering noninfectious virus containing partially double-stranded DNA circles and that particles containing supercoiled HBV DNA may represent infectious hepatitis B virus. PMID- 6286139 TI - Attenuation in the control of SV40 gene expression. AB - Nuclei and viral transcriptional complexes were prepared from cells infected with simian virus 40 and incubated in vitro in the presence of alpha- 32P-UTP. The in vitro elongated viral RNA appeared with a peak of 5S in sucrose gradients and hybridized preferentially to a promoter-proximal region of SV40 DNA. Treatment of infected cells with proflavine led to transcription of elongated RNA, while treatment of cells with 5,6-dichloro-1-beta-d-ribofuranosylbenzimidazole, a drug known to enhance premature termination, augmented accumulation of the promoter proximal RNA. The in vitro elongated RNA produced a major band of 93-95 nucleotides in length in acrylamide gel. This RNA was found to map between the major initiation site at nucleotide 243 and nucleotides 335-337. The significance of these observations with respect to the transcription termination signal and the control of SV40 gene expression is discussed. PMID- 6286138 TI - Isolation of a transforming sequence from a human bladder carcinoma cell line. AB - We have isolated the component of human bladder carcinoma cell DNA that is able to transform mouse fibroblasts. The oncogenic sequence was isolated initially from a lambda phage genomic library made from DNA of a transfected mouse cell carrying the human oncogene. A subcloned insert of 6.6 kb that carried transforming activity was amplified in the plasmid vector pBR322. The subcloned oncogene has been used as a sequence probe in Southern blot analyses. The oncogene appears to derive from sequences present in normal cellular DNA. Structural analysis has failed so far to reveal differences between the oncogene and its normal cellular homolog. The oncogene is unrelated to transforming sequences detected in a variety of other types of human tumor cell lines derived from colonic and lung carcinoma and from neuroblastoma. In contrast, the EJ bladder oncogene appears closely related to one that is active in the human T24 bladder carcinoma cell line. The oncogene appears to have undergone little, if any, amplification in several bladder carcinoma cell lines. PMID- 6286140 TI - DNAase I sensitivity and methylation of active versus inactive rRNA genes in xenopus species hybrids. AB - We studied the chromatin structure and methylation of ribosomal RNA genes (rDNA) in hybrids between Xenopus laevis and Xenopus borealis. S1-nuclease protection experiments showed that 97%-98% of the rRNA precursor in hybrid tadpoles was of the X. laevis type. Preferential expression of the laevis rDNA was correlated with its hypersensitivity to DNAase I compared to borealis rDNA. Borealis and laevis rDNAs gave equivalent methylation patterns, however. The results show that hypomethylated sites in the nontranscribed spacer are not sufficient to ensure DNAase I hypersensitivity or transcription of the borealis rDNA. Also, heavy methylation of the transcribed region of laevis rDNA is compatible with its hypersensitivity to DNAase I. The absence of coupling between hypomethylation and DNAase I sensitivity argues against the view that the methylation pattern directly triggers the active chromatin structure, though it does not exclude a less intimate relationship between transcription and DNA hypomethylation. Examination of borealis sperm rDNA showed that hypomethylated sites were present at the same spacer locations as in somatic cells. This contrasts with X. laevis, where hypomethylated sites are detectable in the spacer of somatic rDNA, but not in sperm. Thus the loss of spacer methylation that is seen in early development of X. laevis does not occur in X. borealis. PMID- 6286141 TI - Instability of transposase activity: evidence from bacteriophage mu DNA replication. AB - Transposition of genetic elements involves coupled replication and integration events catalyzed in part by a class of proteins called transposases. We have asked whether the transposase activity of bacteriophage Mu (the Mu A protein) is stable and capable of catalyzing multiple rounds of coupled replication/integration, or whether its continued synthesis is required to maintain Mu DNA replication. Inhibition of protein synthesis during the lytic cycle with chloramphenicol inhibited Mu DNA synthesis with a half-life of approximately 3 min, demonstrating a need for continued protein synthesis to maintain Mu DNA replication. Synthesis of specific Mu-encoded proteins was inhibited by infecting a host carrying a temperature-sensitive suppressor, at permissive temperature, with Mu amber phages, then shifting to nonpermissive temperature. When Aam phages were used, Mu DNA replication was inhibited with kinetics essentially identical to those with chloramphenicol addition; hence, it is likely that continued synthesis of the Mu A protein is required to maintain Mu DNA replication. The data suggest that the activity of the Mu A protein is unstable, and raise the possibility that the Mu A protein and other transposases may be used stoichiometrically rather than catalytically. PMID- 6286143 TI - Cloning yeast telomeres on linear plasmid vectors. AB - We have constructed a linear yeast plasmid by joining fragments from the termini of Tetrahymena ribosomal DNA to a yeast vector. Structural features of the terminus region of the Tetrahymena rDNA plasmid maintained in the yeast linear plasmid include a set of specifically placed single-strand interruptions within the cluster of hexanucleotide (C4A2) repeat units. An artificially constructed hairpin terminus was unable to stabilize a linear plasmid in yeast. The fact that yeast can recognize and use DNA ends from the distantly related organism Tetrahymena suggests that the structural features required for telomere replication and resolution have been highly conserved in evolution. The linear plasmid was used as a vector to clone chromosomal telomeres from yeast. One Tetrahymena end was removed by restriction digestion, and yeast fragments that could function as an end on a linear plasmid were selected. Restriction mapping and hybridization analysis demonstrated that these fragments were yeast telomeres, and suggested that all yeast chromosomes might have a common telomere sequence. Yeast telomeres appear to be similar in structure to the rDNA of Tetrahymena, in which specific nicks or gaps are present within a simple repeated sequence near the terminus of the DNA. PMID- 6286144 TI - An alteration in the phosphorylation of vimentin-type intermediate filaments is associated with mitosis in cultured mammalian cells. AB - Analysis of cultured Chinese hamster ovary (CHO) cells has shown that vimentin exists primarily as two 57,000 dalton isoelectric variants, a nonphosphorylated form and a slightly more acidic phosphorylated form. Similar analyses of CHO cells that were treated with colcemid show the presence of at least two to three additional, more acidic, phosphorylated vimentin isoelectric variants. An increasing 32P-specific activity of these variants suggests that this alteration involves increased phosphorylation. Analysis of 32P-labeled vimentin from colcemid-treated cells indicates that the amount of the additional phosphorylated variants correlates with the accumulation of cells in mitosis. CHO cells enriched in mitotic cells without antimitotic drugs demonstrate the same alteration in the isoelectric focusing pattern of phosphorylated vimentin. When mitotic cells are replated, the amount of additional phosphorylated variants is reduced within 30 min. The data suggest that an alteration in phosphorylated vimentin is temporally related to the alteration in the organization of intermediate filaments in mitotic cells. PMID- 6286142 TI - Recombination within the yeast plasmid 2mu circle is site-specific. AB - The multicopy yeast plasmid, 2mu circle, encodes a specialized recombination system. It contains two regions, each 599 bp in length, that are precise inverted repeats of each other and between which recombination occurs readily. In addition, this recombination requires the product of a 2mu circle gene, designated FLP. By examining the products of FLP-mediated recombination of plasmids containing single insertions within one of the repeated regions, we show that this recombination occurs only at a specific site within the repeat. This result was confirmed from analysis of the ability of plasmids containing various deletions within one of the repeated regions to serve as substrates for FLP mediated recombination. These experiments limit the recombination site to a sequence of less than 65 bp. In addition, by mutational analysis of the recombination potential of a hybrid plasmid containing the entire 2mu circle genome, we have shown that FLP is only the 2mu circle gene necessary for this site-specific recombination. Finally, we describe a sensitive assay for recombination between the repeated sequences of 2mu circle; using it, we demonstrate that even in the absence of FLP gene product, recombination between the repeats occurs at a low but detectable level during meiosis. PMID- 6286145 TI - Association of the synaptic form of acetylcholinesterase with extracellular matrix in cultured mouse muscle cells. AB - Myotubes of a mouse muscle-cell line (C2) synthesize in culture a 16S form of acetylcholinesterase that is normally found only in regions of adult mouse muscle that contain endplates. The 16S enzyme in C2 cell extracts has the properties expected of acetylcholinesterase forms that have a collagen-like tail. In intact cells, the active site of the 16S acetylcholinesterase is protected by a membrane impermeable inhibitor, and this form of the enzyme can be removed by treatment of the cells with collagenase. Thus the enzyme is extracellular. Its extraction by high ionic strength solutions lacking detergent suggests that the 16S form is associated with the extracellular matrix by ionic interactions. Histochemical staining shows focal patches of acetylcholinesterase activity on the cell surface. Collagenase treatment, which removes only the 16S form, abolishes this staining pattern, indicating that the patches consist of the 16S enzyme. We conclude that the 16S enzyme in C2 myotubes occurs in focal patches on the cell surface, where it is associated with the extracellular matrix. PMID- 6286146 TI - A topological model for transcription based on unwinding angle analysis of E. coli RNA polymerase binary, initiation and ternary complexes. AB - DNA unwinding induced by Escherichia coli RNA polymerase is measured for binary, initiation and ternary complexes formed from a unique promoter sequence on simian virus 40 DNA. At 37 degrees C the complexes all have an unwinding angle of 17 +/- 1 base pairs (580 degrees +/- 30 degrees). This unwinding is attributed to an enzyme-stabilized separation of the double helix at the promoter site, which is maintained throughout initiation and elongation. There is no heterogeneity in the unwinding angle of the ternary complex as it progresses down the helical template. The constant DNA unwinding during all phases of transcription leads us to propose the existence of unwindase and rewindase activities on the enzyme that allow it to travel down the helix like a nut on a DNA bolt. During elongation, the unwindase unwinds the DNA helix while the rewindase, lagging by 17 base pairs, displaces the RNA transcript and reseals the helix. Both activities induce a rotation in the DNA double helix relative to the polymerase. The RNA-DNA hybrid also rotates, maintaining both ends of that helix fixed relative to the catalytic and windase sites. Formation of an RNA-DNA hybrid which spans the distal end of the DNA unwound region is proposed as a possible mechanism for polymerase pausing and termination. This model requires that the polymerase direct the transcript past the noncoding DNA strand. Pausing occurs 16-20 nucleotides downstream from the centers of appropriately sized dyad symmetry elements. PMID- 6286147 TI - Molecular interactions in human T-cell-mediated cytotoxicity to Epstein-Barr virus. I. Blocking of effector cell function by monoclonal antibody OKT3. PMID- 6286148 TI - Serum and cell-mediated viral-specific delayed cutaneous basophil reactions during cytomegalovirus infection of guinea pigs. PMID- 6286149 TI - Kinetic analysis of superoxide anion production by activated and resident murine peritoneal macrophages. PMID- 6286150 TI - Suppressor T cells activated by autologous B-cell derived lines inhibit blastogenic responses by peripheral lymphocytes to mitogens or autologous lymphoblastoid cell lines. PMID- 6286151 TI - Effect of human adenovirus on antibody-dependent cellular cytotoxicity (ADCC) in chickens. PMID- 6286152 TI - Production of superoxide, hydrogen peroxide and hydroxyl radicals by phagocytic cells: a cause of chronic inflammatory disease? PMID- 6286153 TI - High yield preparation of isolated human adult hepatocytes by enzymatic perfusion of the liver. PMID- 6286154 TI - [Oxytocin receptors and their importance in the regulation of labor]. PMID- 6286155 TI - Structure-activity relationships of chlorinated benzenes as inducers of different forms of cytochrome P-450 in rat liver. AB - Hexachlorobenzene (HCB) produced increases in ethoxyresorufin (ERR) O-deethylase, aryl hydrocarbon hydroxylase (AHH) and aminopyrine N-demethylase activities in rat liver phenobarbital which were intermediate between those produced by phenobarbital and 3,4-benzpyrene (BP), alpha-Naphthoflavone (ANF) selectivity inhibited ERR activity in BP and HCB-induced microsomes (94% and 88%). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of liver microsomes indicated that HCB did not produce a detectable increase in a polypeptide with electrophoretic properties similar to those of purified cytochrome P-448 (Mr = 56,000). However, HCB did induce a polypeptide with Mr = 53,000 corresponding to one of two polypeptide bands induced by BP. This polypeptide may represent a second form of cytochrome P-448. Purification of HCB to remove possible dibenzo-p dioxin impurities did not alter the 'mixed-type' induction produced by HCB. In contrast to HCB, all other chlorinated benzenes tested resembled phenobarbital as inducers. PMID- 6286156 TI - Induction and repair of DNA strand breaks in cultured human fibroblasts exposed to various phenols and dihydrodiols of benzo[a]pyrene. AB - Cultured human fibroblasts from healthy donors were incubated for 30 min with nine different benzo[a]pyrene (BP) derivatives in the presence or absence of liver microsomes from 3-methylcholanthrene treated rats. The induction and repair of DNA strand breaks were analysed by alkaline unwinding and separation of double and single stranded DNA (SS-DNA) by hydroxylapatite chromatography immediately after the incubation or at various times after the treatment. In the absence of microsomes DNA stand breaks were detected in fibroblasts exposed to 30 microM of each of the six BP phenols (1-, 2-, 3-, 7-, 9- or 11-OH-BP) and the three BP dihydrodiols (BP-4,5-, BP-7,8- or BP-9,10-dihydrodiol). After removal of the BP derivatives from the medium the DNA strand breaks disappeared within 24 h. alpha Naphthoflavone (alpha-NF) caused a decrease in the induction of strand breaks by 1-, 3- and 9-OH-BP but did not affect the induction of strand breaks in cells exposed to BP-7,8-dihydrodiol. In the presence of microsomes DNA strand breaks were found after exposure to 30 microM of each of the six BP phenols (1-, 2-, 3-, 7-, 9- or 11-OH-BP), as well as BP-7,8- and 9,10-dihydrodiol. In contrast BP-4,5 dihydrodiol did not induce strand breaks under these conditions. The induction of strand breaks by BP-7,8-dihydrodiol was enhanced in the presence of cytosine-1 beta-D-arabinofuranoside (AraC). In all cases the DNA strand breaks had disappeared 24 h after removal of the BP derivatives and microsomes except after treatment with BP-7,8-dihydrodiol. PMID- 6286158 TI - Studies on the mechanism of toxicity of the mycotoxin, sporidesmin. I. Generation of superoxide radical by sporidesmin. AB - Sporidesmin (SDMS2), the mycotoxin responsible for 'facial eczema' in ruminants, contains a disulphide group which appears to be intimately involved in its toxic action. The reduced (dithiol) form of sporidesmin has been shown readily to undergo autoxidation in vitro in a reaction which generates superoxide radical (O2-). The autoxidation reaction, which takes place over a wide pH range, is strongly catalysed by trace amounts of copper, although the reaction was inhibited at high concentrations of this metal. Inhibition of the autooxidation of reduced sporidesmin (SDM(SH)2) was also observed in the presence of nickel, cobalt and manganese. Superoxide radical is also generated from SDMS2 itself in a cyclic reduction/autoxidation reaction with glutathione and other thiols; in view of the known toxicity of superoxide and its derivatives, it is suggested that oxygen-free-radicals may be involved in the initiation of the deleterious effects of the mycotoxin. PMID- 6286159 TI - Studies on deoxynucleic acids and related compounds. VI. Synthesis of oligodeoxyribonucleotides containing recognition sequences for restriction endonucleases. PMID- 6286157 TI - Species differences in the biochemical properties of liver microsomal arylamine and arylamide n-hydroxylases. AB - Cytochrome P-448 dependent microsomal N-hydroxylases are key enzymes in the metabolic activation of both arylamides and arylamines. Using 2 acetylaminofluorene (2-AAF) and 2-aminofluorene (2-AF) as substrates, the present report compares the biochemical properties of rat, hamster and mouse liver N hydroxylases. There are marked species differences both in terms of the affinity for the two substrates and in terms of maximum velocity of the enzymes. The rat and hamster liver arylamide N-hydroxylases are induced by pretreatment with 2-AAF which also significantly increases their affinity for the substrate. In mouse liver neither arylamide nor arylamine N-hydroxylases are modified or induced. With 2-AF as substrate, arylamide treatment never enhances N-hydroxylation but it reduces the Km-value of the rat and hamster liver enzymes. Among the effectors tested in vitro, 3-methylcholanthrene (3-MC), 7,8-benzoflavone (BF), benzo[a]pyrene (B[a]P) and miconazole (MN) inhibit hepatic arylamide N hydroxylase in the submicromolar range. Harman (H) and paraoxon (PX) act in a dose-dependent manner in the micromolar range and metyrapone (MP) is not an inhibitor even at 50-microM concentration. Among the position isomers, 1- and 3 AAF are inhibitors of the N-hydroxylating enzymes whereas 4-AAF is not. PMID- 6286160 TI - [Endemic goiter in the Man region (Ivory Coast)]. AB - An epidemiological study involving 616 subjects carried out in the Man region, Cote-d'Ivoire, showed endemic goitre in 54,4% of the group overall and in 80 % of females. Thyroid function of subjects chosen at random in the goitrous and nongoitrous group (105 G and 71 NG respectively) was identical. Compared with a French control group T4 was significantly lower but T3 and TSH were higher, which represents a high TBG level with low iodine elimination. High levels of TSH are not necessarily associated with low T3 or T4 levels, or with clinical signs of a hypothyroid condition. They are probably the result of a transient reaction. T4 TSH and T3-TSH correlations for NG subjects were (--0,40) but the T3-TSH correlation for the G subjects was significantly lower (--0,23). These differences could be explained by the simultaneous high levels of T3 and TSH more frequently encountered in G subjects and by the fact that serious hypothyroid conditions are more frequent in the NG group (6 out of 9). The percentage of hypothyroid conditions is high in both clinical and biological investigations. No anti-thyroglobulin antibodies were found in any of the hypothyroid cases with goitre. The histograms illustrate the clinical and biological heterogeneity of a homogeneous non-medicalised population. PMID- 6286161 TI - [Discussion of bioclinical data obtained in C.H.U. of Abidjan for marasmic children]. PMID- 6286162 TI - [Control of the quality of water in the framework of a national rural water program on the Ivory Coast. Preliminary results]. AB - In the framework of a National Hydraulic Village Program (NHVP) which aim is to equip each village of the Ivory Coast with wells or drillings, a water quality control service was designed, directly on the field, at the Health Rural Sector Bases. The standards of drinkability were revised according to the NHVP constraints. The technics used were adapted to local conditions. Sanitary education was part of the technics used by the field agents. 1,553 wells were analysed by the end of 1978, 17.7% of which were "polluted". Pollution was transient without treatment in 68% of the above cases. The most polluted cases had a rate of fecal coliforms a 1,000 time inferior to the average traditional wells. The drillings were less polluted (10%) than the large diameter wells (24%): in the latter there was a direct correlation between rain fall and pollution. The technical visits by the agents in the villages represented privileged moments of sensibilisation and education of the villagers. PMID- 6286163 TI - Intracellular injection of calcium ion and EGTA into the hamster submandibular ganglion cell. PMID- 6286164 TI - The spread of hepatitis A virus in connection with hepatitis cases in children's communities. AB - In order to investigate the characteristics of hepatitis A virus excretion, 372 children in semi-closed day-care centres were examined at the time of the seasonal increase in hepatitis incidence. Enzyme immunoassay and immune electron microscopy were used to detect virus in faeces. Ten communities were surveyed and, for the purposes of this paper, were divided into four groups on the basis of the observed pattern of virus excretion. The first group consisted of 4 communities with one hepatitis case in each; in this group, there was a relatively low rate of virus excretion throughout the study. The second group comprised 2 communities with one or two hepatitis cases; this group was characterized by an increase in the rate of virus excretion some time after isolation of the patient. In the communities of the third group, there were numerous hepatitis cases and a high rate of virus excretion during the first weeks of observation, followed by a significant decrease. There were no hepatitis cases in the fourth group, but some hepatitis A virus shedding was detected.The ratio of the number of cases to the number of virus carriers was 1:11.4 in the first two groups, and 1:3.4 in the third. Administration of normal serum immunoglobulin did not significantly affect the rate of HAV excretion. The cause of the differences in shedding of hepatitis A virus in the communities is unknown, but may possibly be related to the size of the immune section of the population. PMID- 6286165 TI - Primary hepatocellular carcinoma. Present state of the disease and prospects for the future. AB - Primary hepatocellular cancer is a common and virulent malignancy found all over the world. New insights into this disease are being provided by evaluation of the impact of epidemiologic data (hepatitis viral exposure, race, sex, HLA typing, and environmental factors). An enhanced appreciation of the biochemical modulators of hepatocellular cancer metabolism is now also possible. The limited success of current therapies is far from satisfactory, but prospects for the future seem more varied and promising than ever before. PMID- 6286166 TI - Beta-endorphins in labor. PMID- 6286167 TI - Measurement of circulating vitamin D in man. AB - An assay for vitamin D consisting of high-pressure liquid chromatography (HPLC) and ultraviolet (UV) absorbance detection has been developed and used to measure circulating vitamin D concentrations in human subjects during summer and winter and after deliberate exposure to ultraviolet radiation. Extracts of 2-4 ml of serum were initially fractionated on silica Sep-Pak cartridges followed by reverse-phase HPLC and finally quantitated by UV-absorbance during straight-phase HPLC. Using these methods, we determined the normal range for circulating vitamin D in Boston subjects to be less than 0.5 ng/ml to 25 ng/ml (n = 30); subjects sampled during summer months had higher concentrations of vitamin D than those sampled during winter months. In subjects exposed to a single quantitative dose of ultraviolet radiation (UVR), large transient increases in circulating vitamin D3 were observed. Concentrations rose 30-50 fold over the first days after exposure before returning to basal levels by one week. PMID- 6286168 TI - Capillary gas chromatographic method for the analysis of lignans in human urine. AB - We describe a capillary column gas chromatographic (GC) method for the analysis of lignans in urine. Lignans are excreted as mono-glucuronides which are first extracted on a small reversed-phase cartridge of octadecylsilane bonded silica (Sep-Pak C18) and thereafter isolated by anion exchange chromatography on DEAE Sephadex A-25, prepared in the acetate form. Lignan mono-glucuronides are enzymatically hydrolysed and re-extracted on a Sep-Pak C18 cartridge. Quantification is carried out by capillary column GC of the trimethylsilyl ether derivatives. The specificity of the method was checked by GC/MS and the intra assay coefficient of variation (CV) for the two lignans, enterolactone (trans-2,3 bis(3-hydroxybenzyl)butyrolactone) and enterodiol (2.3-bis(3-hydroxybenzyl)butane 1,4-diol) varied between 5 and 8%. Some values for the excretion of these lignans by normal men and women are presented. PMID- 6286170 TI - Effect of halothane on adenylate kinase in porcine malignant hyperpyrexia. AB - The effects of halothane on adenylate kinase activity in porcine muscle have been examined. No abnormality in malignant hyperpyrexia susceptible muscle was found. At clinical concentrations of halothane only slight inhibition of adenylate kinase activity was observed. The inhibition increased with increasing concentrations of halothane and with decreasing concentrations of the substrates AMP or ADP. The inhibition was similar in both malignant hyperpyrexia susceptible and control muscle. It seems unlikely that adenylate kinase is involved directly in triggering malignant hyperpyrexia. PMID- 6286169 TI - Immunological similarity of NCA (non-specific cross-reacting antigen) in feces with alpha 1-acid glycoprotein. AB - We have recently suggested that carcinoembryonic antigen (CEA) may contain alpha 1-acid glycoprotein (AG) antigenic determinant. In the present work we examined a protein with CEA-like activity in the feces of healthy subjects (NCA) for immunological cross-reactivity with AG. When the perchloric acid extract of feces was fractionated on a Sephadex G-200, two fractions (large and small molecular weight) were obtained. The large molecular weight fraction had higher CEA activity than the small one. The perchloric acid (PCA) extract of feces was subjected to affinity chromatography using anti-CEA bound to Sepharose, and the bound protein was labelled with 125I, and then fractionated on a Sephadex G-200 column. Two radioactive peaks, Peak 1 corresponding to an approximate Mr of 180 000 and Peak 2, corresponding to an approximate Mr of 60 000 were found. Both peaks showed immunoreactivity with either anti-CEA or anti-AG. This experiment suggests the presence of two kinds of CEA-reactive proteins in feces: one which may be a big protein with immunological similarity to AG and a second which appears to be a hydrolysed fragment of this protein. PMID- 6286171 TI - A simple, sensitive and rapid method for the estimation of cyclic AMP in urine using luciferase luminescence. PMID- 6286172 TI - Failure of FSH to influence aromatization in human adipose tissue. AB - Since gonadotrophins are known to influence aromatase activity in gonadal tissue, an attempt was made to correlate serum gonadotrophin concentrations with the activity of aromatase enzyme in adipose tissue. No relationship between in-vitro measurements of adipose tissue aromatase activity and FSH or LH levels in serum was found. Four of five women studied with premature ovarian failure had low levels of aromatase activity despite having postmenopausal levels of serum gonadotrophins. In addition, FSH receptors could not be demonstrated on while adipose tissue, isolated fat cells or stromal vascular tissue. PMID- 6286173 TI - Functional roles of monoaminergic pathways to sympathetic preganglionic neurons. AB - Despite considerable progress in mapping the central monoaminergic pathways to sympathetic preganglionic neurons in the spinal cord, the respective functional roles of these pathways have not been resolved. Evidence for both excitation and inhibition has been advanced for each of the three monoamines, serotonin, norepinephrine, and epinephrine. Our previous studies on spinal sympathetic pathways to sympathetic preganglionic neurons support the prevailing opinion that serotonin pathways are inhibitory but did not satisfactorily resolve the functional role of the norepinephrine pathways. However, more recent studies showing that intraspinally evoked sympathetic discharges were rapidly and markedly enhanced by phosphodiesterase inhibitors and that this effect was prevented by clonidine have led to formulation of a coherent hypothesis which accommodates much of the conflicting evidence regarding norepinephrine. In addition, evidence for the role of the epinephrine pathways has been obtained by using a selective inhibitor of epinephrine synthesis. The results of these recent studies complement our previous results and suggest that the excitability of sympathetic preganglionic neurons is regulated by excitatory norepinephrine pathways and inhibitory epinephrine pathways that activate or suppress adenylate cyclase to control intraneuronal levels of cyclic AMP. PMID- 6286174 TI - Studies of the functional significance of angiotensin II receptors in the rat olfactory bulb: evidence for alterations in normal fluid intake. AB - The effects of continuous infusion of angiotensin II (ANG II, 1 microgram hr-1) into the olfactory bulb (O.B.) were studied on rats with chronically indwelling intracerebral cannulae. Chronic infusion (ALZA minipump system) of ANG II into the O.B. elicited a moderate dipsogenesis as compared to saline infused animals. The increased drinking occurred only during the dark phase (6 pm - 6 am) and appeared to be associated with food intake, which also occurred during this time. Removal of the food or disconnection of the minipump reduced water consumption to comparable levels in both groups. Acute injection of ANG II (250 ng) into the O.B. failed to elicit drinking. Since the olfactory bulb has a high concentration of ANG II receptors, these experiments suggest that ANG II may interact with the O.B. to facilitate the drinking associated with food intake. PMID- 6286175 TI - Alpha and beta adrenergic receptor activity in circulating blood cells of patients with idiopathic orthostatic hypotension and pheochromocytoma. PMID- 6286177 TI - Adrenergic receptors in human and experimental pheochromocytoma. AB - Radioligand binding to adrenergic receptors has become an important technique for identification and quantitation of these receptors. In this article we review our recent findings using subtype-selective radioligands for studying alpha 1 (e.g., [3H]prazosin) and alpha 2 (e.g. [3H]yohimbine) receptors in human platelets and rat renal cortical membranes, and we report new results on changes in adrenergic receptors of patients and animals with pheochromocytoma. Five patients with pheochromocytoma were tested prior to surgery. The number of alpha 2-adrenergic receptors on platelets from these patients and the affinity of these receptors for [3H]yohimbine were similar to values for control subjects. In rats with a transplantable pheochromocytoma we found a 50-fold increase in plasma norepinephrine levels. The number of renal cortical beta-adrenergic receptors (identified using [125I]iodohydroxybenzylpindolol) and alpha 1-adrenergic receptors were decreased while the number of alpha 2-adrenergic receptors did not change. Receptor affinities for radioligands were unaltered in animals with pheochromocytoma. These findings indicate that pheochromocytoma is associated with selective decreases in the number of renal beta- and alpha 1-adrenergic receptors without changing alpha 2-adrenergic receptor number or affinity. Furthermore, the number and affinity of alpha 2-adrenergic receptors are not altered in platelets of patients with pheochromocytoma. We conclude that down regulation (an agonist-mediated decrease in receptor number) occurs in vivo for some, but not all, types of adrenergic receptors. PMID- 6286178 TI - [Gn . RH receptors in the pituitary and ovaries (author's transl)]. PMID- 6286176 TI - Dietary sodium and renal alpha 2-adrenergic receptors in Dahl hypertensive rats. AB - Dahl sodium-sensitive and resistant rats were fed low and high sodium diets. Renal plasma membrane alpha 2-adrenergic receptor concentrations were greater (p less than .05) in sensitive than in resistant rats while ingesting a low sodium diet. On a high sodium diet the alpha 2 receptor concentration increased (p less than .01) in sensitive but not in resistant rats. Since alpha 2 receptors are located in proximal tubules at sites where norepinephrine induces sodium reabsorption, we postulate that this genetically determined abnormal receptor regulation could lead to exaggerated sodium retention and possibly high blood pressure. PMID- 6286179 TI - [Endorphins and gonadotropin secretion (author's transl)]. PMID- 6286182 TI - [LH receptor binding inhibitor (LHRBI) (author's transl)]. PMID- 6286180 TI - [The central action of catecholestrogens (author's transl)]. PMID- 6286181 TI - [Gonadotropin receptor (author's transl)]. PMID- 6286183 TI - [Significance of the urinary 3',5'-cyclic adenosine monophosphate (cAMP) measurement for the diagnosis of parathyroid disorders (author's transl)]. PMID- 6286184 TI - Pagetoid reticulosis. PMID- 6286185 TI - Pox virus infection in Darier's disease. PMID- 6286186 TI - Transcutaneous sympathetic stimulation: effects of autonomic nervous function. AB - In recent years the use of transcutaneous electrical nerve stimulation (TENS) for the treatment of painful disorders has been widely adopted. Minor side effects such as skin irritation are recognised but with appropriate safeguards the technique is considered free from permanent sequelae. This study examines the effects of TENS on hand skin temperature in 6 normal volunteers and 3 patients with alcoholic peripheral neuropathy. Changes of skin temperature were registered using thermocouples and a thermovision camera. Digital artery pulsation was recorded using the Doppler ultrasound technique. Stimulation at 5 to 10 per second of the ulnar nerve at the elbow and digital nerves in the fingers was maintained for 1 to 4 minutes. Profound cooling of the skin and decrease in arterial pulsation was observed and usually persisted for several minutes after cessation of stimulation. Digital arterial changes were not seen in the 3 patients with neuropathy. The possible physiological basis of the observations is discussed and the implications for TENS therapy considered. PMID- 6286187 TI - Neuropathology of the cortical lesions of the Parkinsonian-dementia (PD) complex of Guam. AB - The Guam amyotrophic lateral sclerosis and Parkinsonian-dementia (ALS and PD) complex still presents a challenge to research. Previous papers have highlighted the lesions in the hippocampus and basal nuclei to explain the bizarre clinical syndrome. In our study of 20 to 62 cases of PD with or without ALS, significant widespread and often severe lesions were found throughout the entire cerebral cortex. These changes included variable degrees of nerve cell loss, spongy change, gliosis, increased lipofuscin content of nerve cells, the presence of granulovacuolar bodies and neurofibrillary tangles within neurones and, less frequently, Hirano bodies. The lesions show a characteristic topographical distribution. The global involvement of the cerebral cortex correlated well with the loss of higher mental function which was present in the patients. PMID- 6286189 TI - Effect of prostaglandins and aspirin on active E-rosette formation in patients with multiple sclerosis. PMID- 6286188 TI - Detection of virus-specific IgA antibodies in serum of kidney transplant patients with recurrent cytomegalovirus infection by enzymeimmuno and radioimmunoassay techniques. AB - The feasibility of using human cytomegalovirus (CMV)-specific IgA antibody determinations as a signal for early detection of recurrent CMV infections in eight renal transplant recipients was analyzed. Solid phase radioimmunoassay (RIA), enzyme-liked immunosorbent assay (ELISA) and immunoperoxidase assay (IPA) techniques were used for IgA antibody determinations. In parallel, IgG antibodies to CMV were studied by immunoperoxidase assay. A significant rise of CMV-specific IgG antibody titre was observed in all of these patients between 5 and 53 weeks post-transplantation. CMV-specific IgA antibody production was detected close to the time a rise in CMV IgG antibody was observed in seven out of eight patients studied by RIA and ELISA, and in six out of eight patients studied by IPA. In two patients specific CMV IgA antibodies were detected by all three methods before a significant rise of CMV IgG antibody titre was demonstrated. In these patients CMV IgA was detected by RIA earlier than by ELISA and IPA. The potential application of CMV-specific IgA antibody determination for early detection of recurrent CMV infection in renal transplant patients is discussed. PMID- 6286190 TI - Association of neutrophil chemiluminescence with microbicidal activity. PMID- 6286191 TI - Restoration of impaired immune functions in aging animals. V. Long-term immunopotentiating effects of combined young bone marrow and newborn thymus grafts. PMID- 6286192 TI - The enhancement of immunoglobulin synthesis by human lymphocytes with lithium. PMID- 6286193 TI - Suppressor T-cell function in man: suppression of immunoglobulin production by the direct action of immunoregulatory T cells on the B cell in four separate, distinct systems. PMID- 6286194 TI - Natural killer activity against retrovirus infected cells: dichotomy in NK sensitivity of infected T and B cells. PMID- 6286195 TI - Detection and analysis of inborn and acquired complement abnormalities. PMID- 6286196 TI - Sera from breast cancer patients contain an IgA antibody to a breast cyst fluid component. PMID- 6286198 TI - Detection of immune complexes in polymorphonuclear leukocytes by double immunofluorescence in patients with IgA nephropathy. PMID- 6286197 TI - Immunopathology of mouse hepatitis virus type 3 infection. IV. MHV3-induced immunodepression. PMID- 6286199 TI - Benign neoplasms and "inflammations" of the breast. PMID- 6286200 TI - Ultrasonic detection of breast cancer. PMID- 6286201 TI - Primary treatment of breast cancer. PMID- 6286202 TI - Metastatic pancreatic vipoma: a case report of clinical response following treatment with corticosteroids and actinomycin D. PMID- 6286203 TI - Myositis ossificans: radiologic evaluation of two cases with diagnostic computed tomograms. AB - Although most physicians associated myositis ossificans with recent, acute trauma, only 40%-60% of patients give such a history. The appearance of a soft tissue mass without a clear history of trauma may suggest a diagnosis of sarcoma, especially because results of a biopsy of the central portion of an area of myositis ossificans may yield immature, undifferentiated tissue resembling a sarcoma. Pain and rapid growth of a mass are more usual in myositis ossificans than in sarcomas, and careful inquiry may reveal stretching injury or chronic trauma associated with normal, vigorous, physical activities. Recognizing the characteristic histologic zoning phenomenon (immature tissue centrally surrounded by more mature tissue and a peripheral shell of benign bone) during the biopsy procedure permits the correct diagnosis of myositis ossificans. Plain radiographs or conventional tomograms may reflect this histologic zoning by demonstrating the typical, mature, outer shell of bone. Although additional radiographic studies are not usually necessary, they may be obtained when the mass is suspected to be a sarcoma. In two patients computed tomographic scans clearly demonstrated well defined, peripheral shells of mature bone, diagnostic of myositis ossificans. PMID- 6286204 TI - Giant cell tumor of tendon sheath developing at the site of tendon laceration. AB - A 32-year-old man suffered sharp trauma to the volar aspect of the wrist. A lacerated tendon found on exploration had been left unrepaired and the skin closed primarily. A few months later the patient presented with a nodule at the same site. Reoperation demonstrated a 2 X 1 cm nodule bridging the gap between the stumps of the previously severed palmaris longus tendon. Pathologic examination of the nodule revealed the typical microscopic appearance of giant cell tumor of tendon sheath (GCTTS) in continuity with the lacerated tendon. The literature regarding the relationship of trauma to the development of GCTTS presents interesting controversy about whether this lesion is reparative or neoplastic. PMID- 6286205 TI - Metastatic disease of bone in orthopedic surgery. PMID- 6286206 TI - Salivary gland imaging. PMID- 6286208 TI - Optimal conditions for assay of cytochrome-c-oxidase activity in human skeletal muscle tissue. AB - Conditions for the assay of cytochrome-c-oxidase in human skeletal muscle tissue were studied, including an evaluation of the optimal conditions during the in vitro assay as well as the optimal conditions for storage and treatment of the tissue before the assay. The activity of cytochrome-c-oxidase was assayed polarographically with a Clark oxygen electrode. Optimal oxygen consumption rates were obtained at a cytochrome-c-concentration above 0.2 mmol/l, in the presence of TMPD, 2.2 mmol/l and ascorbic acid, 4.4 mmol/l as reducing agents. Enzyme proportionality was obtained after correction of the oxygen consumption rates for a blank reaction according to one of three alternatives presented. Variations in the homogenizing technique and the degree of dilution of the homogenate (1:11 1:88) had only moderate effects on the enzyme activity. Optimal storage conditions were evaluated by comparing the enzyme activities measured in fresh muscles, frozen muscles and homogenates prepared from fresh muscles. During all storage conditions significantly higher activities were obtained when sucrose buffer (0.25 mol/l) was used as homogenizing medium as compared to phosphate buffer (0.1 mol/l). Freezing and thawing of the muscle tissue before the assay caused an average decrease in the enzyme activity of 50% (P less than 0.005) as compared to the activity obtained in fresh tissue. This untoward effect of the freezing and thawing was reduced when the enzyme activity was analysed in frozen homogenates prepared from fresh muscles. Thus, an average decrease in the enzyme activity of 15% (P less than 0.05) was found in frozen homogenates prepared in sucrose buffer as compared to the activity obtained in the fresh tissue. These findings emphasize the importance of evaluating the effects of the tissue treatment for comparative studies of maximum enzyme activities in vitro. PMID- 6286207 TI - The response of plasma immunoreactive adrenocorticotropin, beta-endorphin/beta lipotropin, gamma-lipotropin and cortisol to experimentally induced pain in normal subjects. AB - 1. We examined the effect of ischaemic pain and sustained isometric muscle contraction on plasma immunoreactive gamma-lipotropin (gamma LPH), beta endorphin/beta-lipotropin (beta END/beta LPH) and corticotropin (ACTH), which are all synthesized from a common precursor (pro-opiocortin), and plasma cortisol in 10 normal subjects. 2. Experimental pain was produced by inflation to 250 mmHg of a sphygmomanometer cuff, placed above the elbow of the 'dominant' arm, after which the subject squeezed a hand dynamometer, loaded to 12 kg, 20 times at 2 s intervals. Blood was drawn before, after 5 and 10 min of pain, and 30 min after release of the cuff. In a control session, the subjects were asked to squeeze the handgrip alone for 5 min at 30% of their maximum strength, a procedure which elevates the blood pressure without causing pain. 3. One subject had unexplained high (30--71 pmol/l) baseline peptide concentrations. Baseline values for the nine other subjects were: ACTH, 7.3 +/- 1.9 pmol/l (mean +/- SEM); gamma LPH, 18.6 +/- 1.0 pmol/l; beta END/beta LPH, 10.0 +/- 1.1 pmol/l; cortisol, 599 +/- 55 nmol/l. Neither procedure significantly increased the plasma concentration of ACTH or any other peptide, whereas plasma cortisol was significantly increased at both 5 min and 10 min. Plasma ACTH was positively correlated with plasma gamma LPH (r = 0.701; P less than 0.001), beta END/beta LPH (r = 0.970; P less than 0.001) and plasma cortisol (r = 0.758; P less than 0.05). 4. The present study demonstrates that, in normal man, plasma endorphins do not change with experimental ischaemic pain. The rise in plasma cortisol without concomitant rise in ACTH is not explained, but suggests the action of some other agent at the level of the adrenal cortex. PMID- 6286209 TI - Endocrine assays in the monitoring of pregnancy. PMID- 6286210 TI - Perinatal nonbacterial infections. PMID- 6286211 TI - Cellular and molecular aspects of amoeboid movement. PMID- 6286212 TI - Autoregulatory control of the expression of alpha- and beta-tubulins: implications for microtubule assembly. PMID- 6286213 TI - Immunolabeling studies of cytoskeletal association in cultured cells. PMID- 6286214 TI - Actin multigene family of Dictyostelium. PMID- 6286215 TI - Approaches for isolating genes encoding the cytoskeletal proteins and for reintroducing the genes into cultured cells. PMID- 6286216 TI - Actin gelation and structure of cortical cytoplasm. PMID- 6286217 TI - Actinogelin: a Ca++-sensitive regulatory protein of microfilament organization. PMID- 6286218 TI - Partial sequence and turnover of rat liver gap junction protein. PMID- 6286219 TI - Involvement of vinculin in contact-induced cytoskeletal interactions. PMID- 6286220 TI - Recycling of cell-surface receptors: observations from the LDL receptor system. PMID- 6286221 TI - Synthesis and assembly of the vesicular stomatitis virus glycoprotein. PMID- 6286223 TI - Are lysosomes a site of enveloped-virus penetration? PMID- 6286222 TI - Mechanisms for the incorporation of proteins into the plasma membrane. PMID- 6286225 TI - Molecular events in cAMP-mediated reverse transformation. PMID- 6286224 TI - Multifunctional role of calmodulin in biologic processes. PMID- 6286226 TI - Mechanism of transformation by Rous sarcoma virus: events within adhesion plaques. PMID- 6286227 TI - Production of collagenase inhibitor by the growth cartilage of embryonic chick bone: isolation and partial characterization. AB - Production of collagenase and collagenase inhibitors by the explants of epiphyseal, metaphyseal and diaphyseal regions of embryonic chick limbs during development has been investigated. Collagenase-inhibitory activity was first detected in the culture medium of the diaphyseal region of limbs at stage 36 where cartilage matrix erosion began to occur. However, neither active nor latent collagenase was detected in the media of any regions examined. At stage 38 and later, the maximum production of the inhibitory activity was observed in the explants of metaphyseal region (growth cartilage), while collagenase production was only in the diaphyseal region. Two collagenase inhibitors were isolated and purified approximately 150 fold from the culture medium of stage 38 and 43 metaphyseal regions of limbs by anion- and cation-exchange chromatography followed by gel filtration. The inhibitors are cationic proteins with the same molecular weight of approximately 25,000, but slight difference in molecular charge. They are heat-stable and inhibit collagenases from tadpole skin, chick bone and skin and human granulocytes and gelatinases from human granulocytes and chick skin as well as trypsin, but not Clostridium histolyticum collagenase. A possible function of the inhibitors in multistep regulations of the collagen degrading enzyme system at the region of osteo-chondral transition is discussed. PMID- 6286228 TI - Dual effects of type II collagen on the degradation of type I collagen by tadpole collagenase. AB - Kinetic analysis of the degradation of mixtures of type I and type II collagens by tadpole collagenase was performed, using [14C]glycine-labeled type I collagen and [3H]acetylated type II collagen as substrates. The degradation rate of type I collagen was decreased in the initial stage of the reaction, then further slowed down giving a constant rate independent of both the amounts of enzyme and type II collagen. The effect of type II collagen on the degradation of type I collagen is two-fold. First, competitive inhibition manifested only in the initial stage of the reaction due to higher (about 4-fold) affinity of type II collagen for the enzyme than type I collagen. Second, interference in step(s) following the formation of enzyme-substrate complex. PMID- 6286229 TI - Collagen synthesis and degradation in the uterine deciduoma: regulation of collagenase activity by progesterone. PMID- 6286230 TI - Basement membrane research in diabetes mellitus. PMID- 6286231 TI - The connective tissue in scleroderma. PMID- 6286232 TI - Cell-free synthesis of putative type V procollagen chains programmed by Chinese hamster lung cell mRNA. AB - A messenger RNA fraction isolated from cultured Chinese hamster lung (CHL) cells programs in a cell-free system prepared from wheat germ the efficient incorporation of [14C] proline into newly synthesized protein with a significant fraction of the incorporated substrate being digestible with bacterial collagenase. This reaction requires both subcellular fractions, an energy source, and is inhibited by the antibiotic puromycin. The relative amount of collagenase digestible to non-digestible cell-free product depends upon the ratio of CHL mRNA to wheat germ lysate, is not affected by either the Mg2+ or K+ concentrations employed, and under optimal condition, approximately 38% of the total incorporated substrate is collagenase-sensitive. Electrophoresis on SDS polyacrylamide gels of the products programmed by CHL mRNA indicates that the collagenase-digestible material corresponds in size to a procollagen chain with an apparent molecular mass of approximately 170,000 daltons. These studies suggest that the collagen alpha 1 (V) chain is initially synthesized as a precursor procollagen chain and demonstrate that a significant amount of the mRNA in Chinese hamster lung cells codes for this protein. PMID- 6286233 TI - Characterization of the cell free translation products from types I and II procollagen mRNAs. PMID- 6286234 TI - An improved method for the purification of vertebrate prolyl hydroxylase by affinity chromatography. AB - Prolyl hydroxylase was purified from extracts of 13-day old chick embryos by an improved affinity column technique. Prolyl hydroxylase was released from the affinity column by poly-L-proline and was separated from the other proteins and from the poly-L-proline by ion exchange chromatography. This improvement allowed eluates from up to six affinity columns to be pooled and chromatographed in a single step. Furthermore, a major contaminating protein which was difficult to remove from the eluted enzyme using the previous procedure was easily separated by the ion exchange column. The overall advantage of the new technique allows much larger amounts of prolyl hydroxylase to be prepared at a single time than was previously possible. PMID- 6286235 TI - Comparable content of hydroxylysine-linked glycosides in subcomponents C1q of the first component of human, bovine and mouse complement. AB - The hydroxylysine-glycosides in bovine and mouse C1q are directly quantified in parallel with those in human C1q after the alkaline hydrolysis of these molecules. Human, bovine and mouse C1q contain 68.3, 66.3 and 64.0 hydroxylysine galactosylglucose residues in each of these molecules respectively. Only human C1q contains 2.5 residues of hydroxylysine-galactose per molecule, and both of bovine and mouse C1q contain no detectable hydroxylysine-monosaccharides in their molecules. The percentage of hydroxylysine residues glycosylated to total hydroxylysine residues in each of these molecules is calculated to be 86.4, 92.0 and 95.1% for human, bovine and mouse C1q respectively and is comparable with each other. The percentages of hydroxylysine residues resistant to periodate oxidation to total hydroxylysine residues in these molecules were 61.1, 65.3 and 74.3% for human, bovine and mouse C1q respectively and were significantly lower than those estimated by the direct quantification of hydroxylysine-glycosides after the alkaline hydrolysis of these molecules. PMID- 6286237 TI - Preparation of a monospecific antibody to purified rabbit synovial fibroblast collagenase. AB - Collagenase produced by monolayer cultures of rabbit synovial fibroblasts was purified from serum-free culture medium. Antiserum to the pure preparation was raised in sheep. The IgG fraction has been isolated and characterized and shown to be specific for collagenase. The antibody is capable of precipitating and inhibiting the activity of both latent and activated rabbit synovial fibroblast collagenase. Cross-reactivity with human antigen from rheumatoid synovial cell culture medium is described. PMID- 6286236 TI - Collagenase production by cloned populations of rabbit synovial fibroblasts. AB - Monolayers of rabbit synovial fibroblasts treated experimentally with phorbol myristate acetate produce large amounts of collagenase and prostaglandin E2 and have been a suitable experimental model for the proliferative/destructive lesion of rheumatoid arthritis. We used X-irradiation to prolong the in vitro lifespan of these cells so that cloned populations could be studied. By a number of criteria, X-irradiation did not alter the cells to make them unrepresentative of synovial fibroblasts. With limiting dilution techniques, we simultaneously isolated three clones. These clones were shown to have different growth rates and to produce different amounts of collagenase and prostaglandin E2. Rates of protein synthesis, measured by incorporation of 3H-leucine, were similar for all three clones. Our data support the concept that particular populations of synovial cells may contribute selectively to the joint destruction seen in rheumatoid disease. PMID- 6286238 TI - Type IV collagens' isolation and characterization of 7S collagen from human kidney, liver and lung. AB - 7S collagens were isolated after bacterial collagenase treatment of basement membrane material prepared from the pepsin digest of human kidney, liver and lung. The 7S collagens were purified by combination of molecular sieve and ion exchange chromatography. 7S collagen from each of these sources showed similar amino acid composition, electrophoretic patterns of reduced and unreduced samples. Antibodies raised against human placental 7S collagen in rabbits completely cross-reacted with the 7S collagen preparation obtained from kidney, liver and lung in enzyme-immunoassays. Since 7S collagen in known to be a major cross-linked segment of type IV collagens, the data indicate that collagenous proteins of basement membranes are organized in similar networks in a variety of organs. PMID- 6286239 TI - Segment-long-spacing crystallites, a powerful tool in collagen research. PMID- 6286240 TI - Assay of collagenase activity by a rapid, sensitive, and specific method. AB - A new method for the assay of collagenase activity has been developed, whereby the collagen cleavage products, after initial collagenase digestion, are degraded further by a mixture of trypsin and alpha-chymotrypsin. The degradation products are soluble in TCA and can be conveniently separated from the remaining uncleaved collagen substrate by rapid filtration. The enzyme assay is shown to be reproducible and sensitive, and it lends itself to a convenient and rapid determination of collagenase activity in relatively large numbers of samples. The applicability of this method is demonstrated by the detection of increased collagenase activity in skin fibroblast cultures derived from a patient with recessive dystrophic epidermolysis bullosa. PMID- 6286241 TI - Stimulation of retinoic acid of synthesis and turnover of basement membrane in mouse embryonal carcinoma-derived endoderm cells. AB - The effect of retinoic acid on the synthesis and degradation of basement membrane components by endoderm cells derived from mouse embryonal carcinoma (EC) cells was studied in a serum-free, defined medium. By immunofluorescence these cells accumulate type IV collagen, laminin, and fibronectin after growth in media containing epidermal growth factor (EGF), fibroblast growth factor (FGF), insulin, transferrin, and Pedersen fetuin. Collagen accounted for 2 to 4% of the newly synthesized proteins, of which 90% were found in the culture media. This collagen was identified as Pro-type IV be gel electrophoresis and enzymatic susceptibility. The EC cells preferentially attached to type IV collagen in vitro and such attachment was mediated by laminin. Treatment of EC cells with retinoic acid caused an increased accumulation of collagen (10 to 15% of secreted proteins) and also stimulated the elaboration of latent protease which degraded laminin and type IV collagen. The laminin-degrading activity was plasminogen dependent. The type IV collagen-degrading activity was a metal protease which could be activated by trypsin or plasmin. It is likely that at least part of the laminin degrading activity is plasmin (mediated through plasminogen activator), since highly purified plasmin is shown to degrade native laminin. PMID- 6286243 TI - Elimination. Faecal incontinence. PMID- 6286242 TI - Metabolism of glycerate-2,3-P2--II. Enzymes involved in the glycerate-2,3-P2 metabolism in chicken skeletal muscle. AB - 1. Four enzyme fractions which may be involved in the synthesis and breakdown of glycerate-2,3-P2 have been isolated from extracted skeletal muscle by gel filtration and ion-exchange chromatography. 2. One of the fractions, corresponding to the glycerate-2,3-P2 dependent phosphoglycerate mutase, has been purified to homogeneity. In addition to the main enzymatic activity, it shows intrinsic glycerate-2,3-P2 synthase activity and glycerate-2,3-P2 phosphatase activity stimulable by glycolate-2-P. Its synthase activity represents about 10% of the total synthase activity of the tissue, and its phosphatase activity corresponds to about 60% of the total phosphatase activity. 3. Two of the fractions have glycerate-2,3-P2 synthase, glycerate-2,3-P2 phosphatase and phosphoglycerate mutase activities in a ratio similar to that of the glycerate 2,3-P2 synthase described in mammalian skeletal muscle. Their synthase activity corresponds to about 90% of the total synthase activity, and their phosphatase activity represents about 1% of the total phosphatase activity of the tissue. 4. The fourth fraction shows only glycerate-2,3-P2 phosphatase activity and represents about 40% of the total activity of the tissue. 5. It is suggested that in chicken skeletal muscle the metabolism of the glycerate-2,3-P2 is regulated in a way similar to that described in mammalian skeletal muscle. PMID- 6286244 TI - Detection of acute myocardial infarction by pyrophosphate 99m Tc scintigraphy. PMID- 6286245 TI - The cytomegalovirus: when and how to look for it. PMID- 6286246 TI - Collagenase in keloid biopsies and fibroblasts. AB - Trypsin-activated collagenase activity was analyzed in media from keloid biopsies as well as keloid fibroblasts using radiolabeled guinea pig skin collagen as a substrate for collagenase digestion. Collagenase activity per tissue dry weight and DNA was similar in keloid and normal dermal biopsies; however, it was significantly elevated in keloid compared to normal dermal fibroblast media. In two paired cell lines where keloid and normal dermal fibroblasts were isolated from the same patient, relative collagen synthesis and trypsin-activated collagenase were increased to a similar extent in the keloid cell lines compared to normals. The biopsy data are in agreement with other findings, and further confirm the hypothesis that increased collagen deposition in keloids does not result from decreased collagen degradation. In vitro, however, the data presented here indicate that in some cell lines increased collagen synthesis may be associated with increased trypsin-activated collagenase. PMID- 6286247 TI - Factors increasing glutaminase flux in hepatocytes. PMID- 6286248 TI - Accelerated recovery from metabolic acidosis. PMID- 6286249 TI - A seroepidemiologic survey of antibody to bluetongue virus in Alabama cattle. AB - Bluetongue (BT) is an insect transmitted viral disease of sheep that often causes mild or inapparent disease but rarely causes severe disease in cattle. Until recently, bluetongue viral infection was believed to be more prevalent in the Western United States, as compared with other regions of the country. However, a national survey for bluetongue antibody and clinical evidence of the disease in the Southeastern United States prompted the present investigation that was designed to determine the serological prevalence of BT virus in Alabama cattle. Results of the study demonstrated that 16% of the samples collected from 1,500 cattle in 64 of the 67 counties were positive. The prevalence of positive cattle in the western part of the State was significantly higher (P less than .001) than the prevalence in the eastern half of the State. On a herd basis, 52% of all herds tested had positive animals. Results of this study suggest that bluetongue infection is more common in the Southeastern United States than previously suspected. PMID- 6286250 TI - Diarrheal condition in dogs associated with viruses antigenically related to feline herpesvirus. AB - Viruses with properties consistent with herpesvirus were isolated from dogs with diarrhea. The viruses were shown to be antigenically related to feline herpesvirus-1 (FHV-1) by virus neutralization tests. It was also observed that a canine herpesvirus (CHV) prototype, D004, and two field isolates from fatal CHV infections in 2-week-old and 6-week-old puppies were neutralized at a low level by antiserum to FHV-1. Reciprocal neutralization tests with CHV antiserum against FHV-1 were negative. These results indicated that viruses related to FHV-1 can infect the dog and that there appears to be uni-directional virus neutralization of CHV by FHV-1 antibody. PMID- 6286251 TI - Electrical network modeling of active membranes of nerves. PMID- 6286252 TI - Narcotic antagonist therapy of the obesity hypoventilation syndrome. AB - A child with respiratory failure and cor pulmonale secondary to the obesity hypoventilation syndrome (OHS) was found to have abnormal beta-endorphin levels in cerebrospinal fluid (CSF) and serum. A single iv dose of 10 microgram/kg of naloxone early in the course of respiratory failure resulted in dramatic improvement which lasted approximately 3 to 4 h. The patient failed to response to progesterone, and because of deteriorating respiratory status a low-dose continuous infusion of naloxone, 2 microgram/kg.h, was begun and gradually increased to 10 microgram/kg.h, during which time there was a dramatic improvement in respiratory status and clinical condition. After 5 days, naloxone infusion was discontinued and progressive respiratory deterioration recurred. The child died of over-whelming sepsis and disseminated intravascular coagulation. PMID- 6286254 TI - Cefotaxime ("Claforan') in routine hospital use. AB - A multi-centre study was carried out in 327 patients with a wide variety of moderate to severe infections. Patients were treated with cefotaxime at the recommended dosages and, in most cases, as the sole antibiotic. The bacteriological eradication rate was 83% while 92% of clinically assessable cases were successful; 3% of cases relapsed and 5% failed to respond. There was a low incidence of side-effects. PMID- 6286253 TI - Microbial ecology and activities in the rumen: part 1. AB - This review describes the progress which has been made during the last 10 to 15 years in the field of rumen microbiology. It is basically an account of new discoveries in the bacteriology, protozoology, biochemistry, and ecology of the rumen microbial population. As such it covers a wide range of subjects including the isolation and properties of methanogenic bacteria, the role of rumen phycomycete fungi, anaerobic energy conservation, and general metabolic aspects of rumen microorganisms. It also attempts, however, to describe and develop new concepts in rumen microbiology. These consist principally of interactions of the microbemicrobe, microbe-food and microbe-host types, and represent the main areas of recent advance in our understanding of the rumen ecosystem. The development of experimental techniques such as chemostat culture and scanning electron microscopy are shown to have been instrumental in progress in these areas. The paper is concluded with an assessment of our present knowledge of the rumen fermentation, based on the degree of success of experiments with gnotobiotic ruminants inoculated with defined flora and in mathematical modeling of the fermentation. The efficacy of chemical manipulation of the fermentation in ruminant is also discussed in this light. PMID- 6286256 TI - Transformation of chick embryo fibroblasts by rous sarcoma virus does not inhibit the assembly of fibronectin into reduction-sensitive dimer and high molecular weight complex. AB - Chick embryo fibroblasts (CEFs) transformed by Rous sarcoma virus synthesize reduced amounts of fibronectin and also shed this protein into the medium more rapidly than do uninfected cells. We wanted to know whether or not the increased fibronectin shedding rate observed in RSV-transformed CEFs could be explained by an inability of fibronectin to form dimers and/or HMW complex. Our studies on normal and RSV-transformed fibroblasts labelled either metabolically or by lactoperoxidase-catalysed iodination indicate that RSV-induced transformation does not alter the subunit structure of either cell-bound or shed fibronectin, nor does it appear to alter the kinetics of conversion of dimeric fibronectin into HMW complex. We conclude that transformation of CEFs by Rous sarcoma virus does not prevent the assembly of fibronectin into dimeric and HMW complex forms and we offer alternative hypotheses for the more rapid shedding of fibronectin protein by these cells. PMID- 6286255 TI - Long-term, low-dose treatment with pivmecillinam alone and in combination with pivampicillin in patients prone to recurrent bacteriuria. AB - The results of three separate studies aimed at evaluating the efficacy of long term, low-dose treatment with pivmecillinam alone, and in combination with pivampicillin, in patients prone to recurrent bacteriuria, are presented. Overall, pivmecillinam and pivmecillinam/pivampicillin appeared to be equally effective, both clinically and bacteriologically, in a group of elderly subjects treated for 6 months. Pivmecillinam was also used successfully for up to 1 year in 10 out of 12 children in whom prophylactic therapy was indicated. The various regimens employed were without any significant toxic effects and were well tolerated. PMID- 6286257 TI - Studies on the biological balance between thromboxanes and prostacyclins in relation to the platelet-vessel wall interaction. PMID- 6286258 TI - [The application of plasma ACTH radioimmunoassay in patients with Shen Yang Xu (author's transl)]. PMID- 6286259 TI - Plant fiber content of selected breakfast cereals. PMID- 6286260 TI - [Activation of energy-independent liver mitochondrial NAD(P)-transhydrogenase by catecholamines, glucagon and cAMP]. PMID- 6286261 TI - [Molecular mechanisms of unbalanced cell death and metabolic mutagenesis]. PMID- 6286262 TI - [Participation of the lymphoid system in the body's resistance to hypoxia]. PMID- 6286263 TI - [DNA-like duplexes containing repetitions. V. Chemical template synthesis of polynucleotides containing 3',5'-pyrophosphate internucleotide bonds]. PMID- 6286264 TI - Endorphinlike drugs. PMID- 6286265 TI - [Virus etiology of Hodgkin's disease]. PMID- 6286266 TI - [Antibiotic treatment of human actinomykoses (author's transl)]. AB - In the complex aetiology of human actinomykoses, various Actinomyces species, especially A. israelii, play the pathogenetically leading role in the mixed flora so typical for these diseases. A number of other microaerophilic and anaerobic bacteria are also part of such mixed flora. Since these "concomitant" bacteria are themselves potentially pathogenic, it is important to have data on antibiotic sensitivity for the entire spectrum of microorganisms, to achieve effective and economic chemotherapy of human actinomykoses. Minimal inhibitory concentrations of antibiotics were determined for Actinomyces and the most important concomitant bacteria from actinomykotic processes: Actinobacillus actinomycetem-commitants, Bacteroides of the Melaninogenicus group, B. fragilis, B. theta-iotaomicron, Fusobacterium nucleatum, Fusobacterium spp. and Propionibacteria. The results indicate that aminopenicillines are the drug of first choice, if in the actinomykoses there are no Bacteroides strains resistant to beta-lactam antibiotics. These tests further indicate that clinically the combination of aminopenicillin and clindamycin, as well as aminopenicillin and metronidazol, are indicated. Minocycline and especially cefoxitin, a beta-lactamase resistant cephalosporin, inhibit with few exceptions all bacteria which are potentially involved in actinomykotic processes. PMID- 6286267 TI - [Beta lactamase inhibitors, a new start of antibiotic therapy]. PMID- 6286268 TI - Morphological and biochemical effects of gangliosides in neuroblastoma cells. PMID- 6286269 TI - Uptake and binding of serotonin by primary cultures of mouse astrocytes. AB - Primary cultures of glia contain a high glial fibrillary acidic protein level and exhibit important glutamine synthetase activity. They take up serotonin via a high affinity carrier-mediated system with a Km of micromolar range. The Km of this transport process does not vary during cell growth or maturation; however, during the last period of morphological change induced by dibutyryl cyclic AMP, an increase in Vmax is observed. Chlorimipramine, fluoxetine and fenfluramine at 10-4 M inhibit this uptake. 3H-5-HT still binds to partially purified astrocytic membranes on a single type of site. During growth, neither KD nor Bmax were modified. During the maturation period, KD decreased to about 50% of its control level. Methysergide inhibits that binding. PMID- 6286271 TI - [Herpes simplex virus and cancer]. PMID- 6286270 TI - Control of cation transport in cultured glial cells by external Ca++: a possible signal in glial-neuronal interaction. AB - In cultured glial cells from chick embryonic brain, both influx and efflux of 42K+ and 22Na+ are dependent on the external Ca++ and concentration ([Ca++]0) between 2 and 0.1 mM although intracellular concentrations of K+ ([K+]i) and Na+ ([Na+]i) do not change. Only a reduction of [Ca++]0 below 0.1 mM results in both a decrease of [K+]i and an increase of [Na+]i. Ouabain significantly decreases the [Ca++]0 sensitivity of uphill cation movements (K+ influx and Na+ efflux), while the [Ca++]0 sensitivity of downhill cation movements (K+ efflux and Na+ influx) is almost not affected by the presence of ouabain. Additionally, a decrease in [Ca++]0 triggers an increase in intracellular concentration of adenosine 3':5'-cyclic monophosphoric acid (cAMP). These findings suggest that changes of [Ca++]0, which take place in vivo in the microenvironment of the glia after neuronal firing, represent a signal in the glial-neuronal interaction controlling cation transport and that this control is achieved by a co-operation between the cAMP-generating and the cation transport system. PMID- 6286272 TI - [The role of the fetus in labor]. PMID- 6286273 TI - Effects of hexavalent chromium in rainbow trout (Salmo gairdneri) after prolonged exposure at two different pH levels. PMID- 6286274 TI - The treatment of duodenal and gastric ulcer with ranitidine--a controlled, multicentre clinical trial. AB - One hundred and seventy seven duodenal ulcer and eighty-one gastric ulcer patients were treated with either ranitidine or placebo in a prospective double blind study. Groups treated with active drug and placebo were comparable and ulcer healing was assessed by weekly endoscopic examinations. Ranitidine was shown to accelerate the rate of spontaneous ulcer healing in both duodenal and gastric ulcer. This acceleration is significant after one week of treatment in duodenal ulcer and from three weeks onwards in gastric ulcer. The ulcer healing rate after three weeks treatment with ranitidine was 85.5% in duodenal ulcer, and 72.5% in gastric ulcer which was significantly higher than in the placebo group where it was 51.7% and 41.5% respectively (P less than 0.005 and P less than 0.01). Treatment with ranitidine for a maximum of six weeks healed 97.1% of duodenal ulcer patients (172 out of 177) and 96.5% of gastric ulcer patients (77 out of 81). Out of total of 258 patients only 9 failed to heal. No adverse events were seen with ranitidine and it would appear free of the side effects that have been seen with cimetidine. In addition there were no significant changes in blood haematology or clinical chemistry parameters. Ranitidine is also given in a more convenient regimen, 150 mg daily as compared to 200 mg three times daily and 400 mg nocte with cimetidine. PMID- 6286275 TI - [Recent trends in psychophysiology of anxiety (author's transl)]. AB - Current views on anxiety and emotions have been profoundly influenced by research data coming from the field of psychoneuroendocrinology. While early work concentrated on identification of hormonal correlates of fear and anxiety and the question of their specificity, most authors now agree that the physiological phenomena are more a manifestation of increased activation of the arousal system than of any emotional state per se, so that the interest has shifted to the identification of the factors responsible for physiological activation. Over the past few years there has been an increasing awareness of the extreme sensitivity of hormonal systems to psychological and social factors. Pituitary-adrenal hormones for instance are maxillary released under conditions of high novelty and uncertainty, with little control ability and no feedback information following attempts to control the situation. Many experimental studies have attempted to clarify the contribution of peripheral hormonal responses to the cognitive evaluation of emotions and to the storage and retrieval of emotional experiences. This field is now rapidly growing with the increasing number of neuropeptides shown to modulate adaptive behaviour. Most significant in terms of pathophysiology is the present concern for understanding the fundamental psychological processes (defense and coping) which enable the subject to dampen the physiological activation normally resulting from internal and external arousing stimuli. Such a knowledge should help to clarify the mechanisms leading from a failure of adaptation to mental and somatic diseases. PMID- 6286276 TI - [Pharmacology of central noradrenergic receptors (author's transl)]. AB - The pharmacological characteristics of adrenergic receptors in the central nervous system (CNS) can be studied using micro-iontophoretic electrophysiological approaches and binding of radiolabeled specific adrenergic receptors ligands. These approaches allowed the identification of different sub classes of adrenergic receptors (alpha 1, alpha 2; beta 1, beta 2) in the CNS. Some of these receptors have a noradrenergic transmission dependent regulation. At the level of cerebral cortex, regulation mechanisms were particularly well studied. Interruption of noradrenergic transmission induces an hypersensitivity of alpha and beta adrenergic receptors. The hypersensitivity is linked to an increase of receptor density, and this has been shown for alpha 1, alpha 2 and beta 1 adrenergic receptors. After repeated injections of antidepressant drugs an hyposensitivity of beta receptors was observed. More precisely these treatments induced a reduction of beta 1 binding sites, without modification of beta 2 binding sites. On the other hand, the alpha 2 binding sites were increased. These regulation processes of alpha 2 receptors appear earlier than those of beta receptors, and this phenomenon could contribute to the late appearance of beta receptors hyposensitivity. In favor of this hypothesis, the reduction of beta binding sites is induced more rapidly after coadministration of alpha 2 receptor antagonist (such as yohimbine) and antidepressant drugs. The regulation mechanisms of these different types of adrenergic receptors could contribute to the delayed action of antidepressant drugs. PMID- 6286277 TI - [Proserotoninergic agents and depression (author's transl)]. AB - The proserotoninergic hypothesis of depression has been for a long time founded on biochemical parameters (brain from suicide patients, CSF and plasma from depressed patients) or more recently on measurement of platelets 5-HT uptake or imipramine binding. New specific proserotoninergic agents confirm this hypothesis since indalpine, a specific 5-HT uptake inhibitor, has a neurochemical and pharmacological profile characteristics of proserotoninergic agents and is effectively antidepressant in human. 5-HT uptake inhibition seems an important property because when we compared two isomers one, a 5-HT uptake inhibitor and at the same time 5-HT releaser, and the other only releaser, we observed that the first has experimental properties similar to indalpine whereas the second has not the classical spectrum of a proserotoninergic agent. PMID- 6286278 TI - Vasopressin resistance induced by low sodium and high mannitol in toad bladder. AB - The natriferic and hydroosmotic responses to vasopressin (ADH) of the isolated toad bladder were studied as a function of the serosal bath NaCl concentration. Bladders were preincubated in Ringer's solution containing equiosmotic substitutions of mannitol, choline chloride, or choline sulfate for NaCl. Their hydroosmotic and natriferic responses to ADH were measured in NaCl Ringer's solution. Bladders previously bathed on their serosal surface with mannitol had a markedly diminished hydroosmotic response, whereas the natriferic response of these bladders was essentially normal. Similarly, preincubation with either choline chloride or sulfate also inhibited the hydroosmotic response, although the decline was substantially less than that with mannitol. Preincubation with mannitol also inhibited the hydroosmotic response to dibutyryl cAMP. Therefore, the isosmotic substitution of serosal bath NaCl with mannitol decreases primarily the hydroosmotic response to ADH at a post-AMP step. The natriferic effect of the hormone remains essentially intact under these conditions. Different compartments (or cell types) appear to regulate the action of ADH on sodium and on water transport in this tissue because of the selective effects of preincubation with low NaCl Ringer's solution on the hydroosmotic and natriferic responses to ADH. PMID- 6286279 TI - The effect of hypoxia on insulin and glucagon secretion in the perfused pancreas of the rat. AB - It has been reported that insulin secretion decreases during hypoxia both in vitro and in vitro, while an increase in glucagon secretion is found only in vivo. The effect of acute hypoxia on the secretion of glucagon and insulin was studied in the perfused rat pancreas. Phentolamine, an alpha-adrenergic blocker, was perfused during the period of hypoxia to elucidate the role of alpha adrenergic stimulation. Sodium ATP and dibutyryl cAMP were also administered to study their effects on insulin and glucagon responses during hypoxia. In the present experiments, insulin secretion was suppressed while glucagon secretion was increased during hypoxia. Phentolamine did not cause any change in insulin of glucagon secretion. When dibutyryl cAMP was added, the increased glucagon secretion was reduced to the basal level, whereas the decreases in insulin secretion were not altered. The addition of sodium ATP reversed the hypoxia induced decrease in insulin and the increase in glucagon secretion. These results suggest that a decrease in ATP production, which leads to impaired cAMP generation, pays a role in, and that alpha-adrenergic stimulation does not participate in the changes in, insulin and glucagon secretion during hypoxia in vitro. PMID- 6286280 TI - Induction of somatogenic receptors in livers of hypersomatotropic rats. AB - Male and female Wistar-Furth rats bearing the pituitary tumor MtT/W15 had serum GH and PRL levels several hundred-fold higher, and immunoreactive somatomedin levels 3-fold higher, than those of controls. The presence of tumor appeared to have no effect on specific binding of radioiodinated bovine GH to liver microsomal membranes in male animals, and to decrease specific binding by 78% in females. However, after desaturating receptors by treatment with 3.2 M MgCl2 for 5 min, bovine GH binding to membranes from tumor-bearing males was 3-fold higher than in normal males, while in tumor-bearing females binding was more than twice as high as in normal females. Competitive binding curves showed the induced receptors to be specific for growth hormones, and thus somatogenic in nature. This study indicates that a high degree of occupancy of GH receptors in hypersomatotropic rat liver does not cause their down-regulation. The mechanism of receptor induction is unknown. PMID- 6286281 TI - Pregnenolone biosynthesis by cultured rat granulosa cells: modulation by follicle stimulating hormone and gonadotropin-releasing hormone. AB - The mechanism by which GnRH inhibits ovarian progesterone production was investigated by studying the GnRH modulation of pregnenolone biosynthesis in cultured rat granulosa cells. Granulosa cells from hypophysectomized, estrogen treated rats were incubated for 2 days with various hormones in vitro. Pregnenolone production was measured in the presence of cyanoketone, which inhibits the conversion of pregnenolone to progesterone. FSH stimulated pregnenolone production in a dose-dependent manner (ED50, 4.47 ng/ml). Concomitant treatment with GnRH resulted in a dose-dependent decrease in FSH stimulated pregnenolone production (ID50, 6.3 x 10(-9) M; maximal decrease, approximately 50%). In contrast, treatment with high doses of GnRH alone stimulated pregnenolone production (ED50, 2.95 x 10(-8) M) reaching a maximal level of about 10% that induced by FSH. Treatment with a GnRH antagonist, [Ac-D Phe1, D-pCl-Phe2, D-Trp3,6]GnRH, did not affect either basal or FSH-stimulated pregnenolone production, but blocked both inhibitory and stimulatory effects of GnRH. The addition of 25-hydroxycholesterol, a soluble substrate for side-chain cleavage enzymes, enhanced FSH-stimulated pregnenolone production, but failed to overcome the inhibitory action of GnRH. GnRH also inhibited progesterone production stimulated by 8-bromo-cAMP and cholera toxin. This action of GnRH, however, was not associated with an inhibition of pregnenolone biosynthesis, but appeared to be due to a preferential increase in the metabolism of progesterone to 20 alpha-hydroxypregn-4-en-3-one. Thus, in addition to the reported GnRH stimulation of progesterone metabolism to 20 alpha-hydroxypregn-4-en-3-one and the GnRH inhibition of FSH-stimulated 3 beta-hydroxysteroid dehydrogenase activity, the present results demonstrate that GnRH also inhibits FSH-stimulated pregnenolone biosynthesis, probably at the side-chain cleavage enzyme step. PMID- 6286282 TI - Binding affinity and biological activity of gonadotropin releasing hormone agonists in isolated pituitary cells. AB - The relationships between binding affinity and the biological potency of eight GnRH agonist analogs were evaluated in isolated rat pituitary cells. For this purpose, binding affinity and biological activity were assayed under similar physiological conditions in medium 199, pH 7.4, and binding affinity was also measured under the standard conditions in hypotonic buffer at low temperature. Under physiological conditions, receptor binding affinity was consistently lower than when measured in the hypotonic Tris buffer usually employed for GnRH receptor studies. In the low temperature binding assay at 0 C, which provided a measure of the affinity constant without degradation, a difference of 20- to 30 fold was observed between native GnRH and its most potent analog, [D-Ser(t Bu)6]des-Gly10-GnRH N-ethylamide. Modifications of the amino acid residues at both positions 6 and 10 of the decapeptide increased the binding affinity of GnRH analogs. When the receptor binding assay was performed at 37 C, the range of the apparent affinity constants was extended up to 60-fold. The affinity constants derived at 37 C were closely correlated with the biological potencies of the individual analogs measured in the same cell system. The effect of temperature on binding affinity was not significantly influenced by peptide metabolism, which was minor in the absence of horse serum from the incubation medium. At the pituitary level, the biological potency of the GnRH agonist analogs is predominantly determined by their higher receptor affinity, and reduced degradation is a less important aspect of the high biological activity of the superagonist analogs. PMID- 6286283 TI - Isoelectric heterogeneity of secreted renin differs after beta-adrenergic stimulation. AB - Renal cortical slices from rat kidney were found to contain and secrete six forms of renin (EC 3.4.00.19). The multiple forms were demonstrated by an isoelectric focusing technique utilizing a stable, steady state pH gradient. The isoelectric points of the six secreted forms had the following pH values: 5.70, 5.55, 5.30, 5.15, 4.9, and 4.8, and represent, respectively, 1%, 62%, 17.5%, 8.5%, 8%, and 3% of the secreted renin. In the presence of isoproterenol (10(-6) M), renin secretion from the renal cortical slices was increased significantly (87%) over control (P less than 0.05). The increased renin secretion was accompanied by a significant change in the relative proportions of the six secreted forms of renin. These results indicate that multiple renin forms are stored and secreted by the rat kidney and that stimuli that increase renin secretion can preferentially increase the secretion of some of the multiple forms. It is hypothesized that isoproterenol stimulates the secretion of stored renin, which appears to be composed of a subset of the multiple renin forms. PMID- 6286284 TI - Forskolin stimulation of thyroid adenylate cyclase and cyclic 3',5'-adenosine monophosphate accumulation. AB - The diterpene forskolin is a potent (100-fold) stimulator of guinea pig thyroid cAMP accumulation with half-maximal activation occurring at 40 microM. Forskolin stimulation is more rapid than that of TSH, attaining a 5-fold increase within 1 min of exposure. The stimulation is also rapidly reversible. The diterpene does not sensitize thyroid cAMP accumulation to TSH, and the concentration yielding half-maximal response is not altered by the presence of low levels of forskolin. At maximally stimulating concentrations, the effects of TSH and forskolin on cAMP accumulation are additive. Forskolin stimulates thyroid adenylate cyclase approximately 10-fold in membranes from several species with half-maximal effects occurring at 3--9 microM through an action on the maximum velocity and not the Km for ATP. The activation of thyroid membranes is readily reversible. Guanyl nucleotides are not required for stimulation by forskolin, and they do not sensitize to forskolin. Moreover, the drug did not sensitize the membrane adenylate cyclase to guanosine 5'-[beta, gamma-imido]triphosphate or to isoproterenol and was equally effective with either Mg++ or Mn++ as the divalent cation. Forskolin stimulation is additive with that of guanyl nucleotides and F-. The site of action of forskolin in the adenylate cyclase complex is uncertain. Data from Bordetella pertussis, testicular, and S-49 lymphoma mutant cyclases suggest that one of the guanyl nucleotide regulatory proteins may be required to promote the forskolin effect. We conclude that forskolin is a useful activator of thyroid adenylate cyclase both in vitro and in intact tissue, which will be useful in elucidating the coupling process of the adenylate cyclase system and in differentiating cAMP-mediated from other forms of activation of the thyroid. PMID- 6286285 TI - beta-Endorphin in hypophyseal portal blood: variations throughout the menstrual cycle. AB - Concentrations of beta-endorphin were measured in the venous effluent of the hypothalamus (hypophyseal portal blood) at various phases of the menstrual cycle and after ovariectomy in rhesus and pigtailed monkeys. In the rhesus, beta endorphin concentrations were high during the mid- to late follicular phase [737 +/- 256 pg/ml (mean +/- SE)] and the luteal phase (1675 +/- 1108) of the menstrual cycle, but were undetectable (less than 133) at menstruation. Concentrations were also high in pigtailed monkeys during stages of the menstrual cycle other than at menstruation (4870 +/- 1090 pg/ml), but undetectable (less than 133) 4--12 months after ovariectomy. These results indicate that beta endorphin concentrations in hypophyseal portal blood are related to menstrual cycle events, probably changes in ovarian steroids; this in turn suggests that beta-endorphin may participate in the ovarian feedback regulation of gonadotropin secretion. PMID- 6286286 TI - The effects of adrenocorticotropin, guanylylimidodiphosphate, dibutyryl adenosine 3',5'-monophosphate, and exogenous substrates on corticosteroid output by ovine fetal adrenal cells at different times in pregnancy. AB - To examine the factors responsible for the changing responsiveness of ovine fetal adrenals during pregnancy, we incubated dispersed adrenal cells obtained from fetal sheep on days 50, 100, and 130 of pregnancy and at term with ACTH (1--24), a GTP analog [guanosine 5(1)-(beta, gamma-imido)triphosphate], dibutyryl cAMP, and potential substrates for corticosteroid production, pregnenolone, progesterone, 17 alpha-hydroxypregnenolone, and 17 alpha-hydroxyprogesterone (17OHP4). We determined the output of cortisol (F), corticosterone (B) and progesterone (P4) by these cells during in vitro incubation. ACTH stimulated F and B output on day 50 and at term, but not in midpregnancy. The mean F to B output ratio for all concentrations of ACTH rose from 3.3 on day 50 to 11.3 at term. The GTP analog stimulated F, B, and P4 output at term, but not on day 100 or 130. Dibutyryl cAMP stimulated F and B output on day 50 and at term and stimulated P4 output at all stages of pregnancy examined. Exogenous 17OHP4 was converted to F at all times in pregnancy. The relative conversion of P4 to F and 17OHP4 to F (conversion quotient) rose from 0.09 on day 100 to 0.80 at term. P4 was converted to B at all times in gestation. At term, formation of F from delta 4 substrates (P4, and 17OHP4) was greater than that from delta 5 substrates (pregnenolone and 17 alpha OH-pregnenolone). We conclude that (1) the output of F in response to ACTH was highest on day 50 and at term pregnancy; 2) the loss of response in midpregnancy may be associated with a defect in GTP and cAMP generation and with reduced 17 alpha-hydroxylase activity; and 3) a term, 17 alpha-hydroxylase appears to be less limiting to F production than in midgestation, but delta 5, 3 beta-hydroxysteroid dehydrogenase may be more so. PMID- 6286287 TI - Estrogenic potency, receptor interactions, and metabolism of catechol estrogens in the immature rat uterus in vitro. PMID- 6286288 TI - Age-related differences in the response of hepatic microsomal glucose-6 phosphatase to triiodothyronine and triamcinolone in the rat. AB - A previous report from this laboratory demonstrated age-related differences in the hormonal regulation of hepatic glucose-6-phosphatase (G-6-Pase). A detailed kinetic analysis of G-6-Pase activity has been performed to distinguish between effects on the microsomal carrier for glucose-6-phosphate and those on the enzyme itself. The maximum velocity (Vmax) was determined in untreated microsomes and microsomes treated with sodium deoxycholate (DOC); the Vmax of the enzyme (VE) was equal to the Vmax in the presence of DOC, and the Vmax of the carrier (VT) was calculated from the Vmax of untreated microsomes and the latency (the activity of DOC-treated microsomes not expressed by the intact preparation). The age-related decrease in G-6-Pase activity was caused by a decrease in the VE. In 3-month-old rats, the VE was increased 2,5-fold by treatment with T3, whereas triamcinolone or the two hormones in combination caused little effect; in 24 month-old animals, the VE was increased 10-fold by T3 and 2-fold by either triamcinolone or the two hormones in combination. In contrast, in 3-month-old animals, the VT was increased 2-fold by triamcinolone, 1.5-fold by T3, and 2-fold by the two hormones in combination; in 24-month-old animals, the VT was increased 3.5-fold by triamcinolone, was not affected by T3, and was increased 1.5-fold by the two hormones in combination. These differences could not be explained by changes in the response of isolated microsomes to sodium deoxycholate or by effects on the energy of activation of G-6-Pase. The results provide detailed evidence for an altered response to either T3 or triamcinolone in hepatocytes from old animals. PMID- 6286289 TI - Temporal characteristics of gonadotropin interaction with rabbit luteal receptors and activation of adenylyl cyclase: comparison to the mode of action of catecholamine receptors. AB - The temporal characteristics of gonadotropin (LH and hCG) and catecholamine interaction with their luteal receptors and activation of adenylyl cyclase were studied in rabbit luteal membranes. studies on hormone-receptor interaction showed that, once bound, [125I]iodo-hCG dissociated from its receptor very slowly. If excess LH was added 30 min after the initiation of [125I]iodo-hCG binding, 85% of the [125I]iodo-hCG bound at 30 min was still bound to the luteal receptors 4.5 h later. The rate of dissociation of [125I]iodo-hCG from its receptor was not altered by 100 microM GTP, 2 mM MgCl2, or GTP plus MgCl2. The slow rate of [125I]iodo-hCG dissociation observed at 30 min was not due to a time dependent change in the hormone-receptor complex, as the dissociation of [125I]iodo-hCG was equally slow 5 min after the initiation of the binding reaction. Studies on the activation of luteal adenylyl cyclase by LH showed that stimulation by 1 microgram/ml ovine LH (oLH) could be prevented but, once initiated, could not be reversed by antiserum to oLH. This indicates that once bound to the rabbit luteal LH receptor, oLH causes persistent activation of rabbit luteal adenylyl cyclase. In contrast, the activation of luteal adenylyl cyclase by 1 microM isoproterenol could be completely reversed by the addition of 50 nM propranolol 5 min after the initiation of the reaction. The inhibitory effect of the propranolol could be completely overcome by the addition of excess isoproterenol, indicating that catecholamine binding to its luteal beta-receptor is readily reversible. Thus, there appears to be a basic difference in the mechanism by which the gonadotropins and catecholamines interact with their receptors and activate the rabbit luteal adenylyl cyclase. PMID- 6286290 TI - Gonadotropin binding and Leydig cell activation in the rat testis in vivo. AB - Testicular LH receptor occupancy and steroidogenic responses were measured in adult male rats after intracardiac injections of [125I]iodo-hCG (0.5--5 x 10(6) cpm) mixed with known amounts of nonradioactive hCG to yield doses ranging from 10 ng to 300 micrograms. Uptake of the hormone by the testis was measured in the whole tissue or the 20,000 x g homogenate, with correction for nonspecific binding in animals injected with a 100-fold excess of unlabeled hCG. The steroidogenic response to hCG was followed by measurements of serum and testicular testosterone. Maximum specific uptake of [125I]iodo-hCG by the testes was observed 4--6 h after hormone injection. Of the specific counts, 80% were recovered in the 20,000 x g pellet of the tissue homogenate. The testicular contents of hCG-binding sites were similar when measured by in vivo occupancy of the receptors and by the in vitro receptor assay, indicating the physiological validity of the receptor measurements in tissue homogenates. Serum and testicular testosterone levels reached a maximum at 1 h, independent of the hCG dose used. When receptor occupancy in vitro after injection of hCG was compared with stimulation of steroidogenesis, a significant (P less than 0.05) 3-fold elevation of serum testosterone was seen when only 0.05% of the receptors were occupied. The maximal testosterone response was reached with 0.8% receptor occupancy. It is concluded that the same number of testicular LH receptors can be occupied by the circulating hormone in vivo and in tissue homogenates in vitro. The spare receptor concept also applied to the in vivo situation, since stimulation of steroidogenesis in the intact animal requires occupancy of only a few receptors per Leydig cell. This may be a general feature of hormonal activation of endocrine target cells in vivo. PMID- 6286291 TI - Effect of prolonged low dose infusion of angiotensin II and aldosterone on rat smooth muscle and adrenal angiotensin II receptors. PMID- 6286292 TI - Specific cytosol binding proteins for 25(OH)D3 and 1.25(OH)2D3 in human breast cancers. AB - Binding proteins for 1.25 (OH) 2D3 were investigated in thirty breast cancers. Human breast cancer was shown to contain specific, high affinity cytosol binding proteins for 1.25 (OH) 2D3 and 25 (OH) D3. The binding protein for 1.25 (OH) 2D3 sedimented at 3.7 S and the binding protein for 25 (OH) D3 at about 6.0 S on sucrose density gradient analysis containing 0.3 M KCl and 1 mM dithiothreitol in buffer. Kd for 1.25 (OH) 2D3 were from 0.1 x 10(-11) M to 7.1 x 10(-11) M measured by Scatchard plots. Competition binding studies indicated that the relative specificity of the binding protein for 1.25 (OH) 2D3 much greater than 25 (OH) D3 greater than 1 alpha (OH) D3, 24,25 (OH)2D3 greater than D3 much greater than Estradiol-17 beta. 1.25 (OH) 2D3 receptor-positive was detected in twenty-eight out of thirty breast cancers. PMID- 6286293 TI - Release of human chorionic gonadotropin (hCG) and its alpha-subunit (hCG-alpha) from perifused human placenta. AB - The release of human chorionic gonadotropin (hCG) and its alpha-subunit (hCG alpha) from the normal human placenta and the effect of some stimulatory agents on their release were studied in vitro using a perfusion system. Each perfusate was assayed for hCG and hCG-alpha in its own homologous radioimmunoassay systems. Both hCG and hCG-alpha were released from the placenta at any stage of gestation in our perfusion system. Much more hCG than hCG-alpha was released from the placenta in early gestation. By comparison, however, hCG-alpha increased gradually with the gestational age. The amount of hCG-alpha released was almost equal to that of hCG in the placenta in the 17th gestational week. After the 22nd gestational week, hCG-alpha was released in larger quantities than hCG, and about 10 times more hCG-alpha than hCG was released from the term placenta. These results were also confirmed by gel filtration of perfusates on a Sephadex G-100 column. hCG-alpha, compared with hCG, was present in excess in gel filtrated perfusates in the last two trimesters. By adding 1 mM dibutyryl cyclic AMP to the perifusion medium, the release of both hCG and hCG-alpha was stimulated significantly. Synthetic luteinizing hormone releasing hormone (LH-RH) at concentrations of 10 ng/ml and 100 ng/ml had no effect, but at a high concentration (1 microgram/ml), LH-RH stimulated the release of them. Moreover, mouse epidermal growth factor (EGF) stimulated not only the release of hCG and hCG-alpha but also their production, because both hCG and hCG-alpha levels rose progressively with the time course in the presence of EGF. The present studies demonstrate that the perifusion system of chorionic tissues is a useful method for investigating the release of hCG and its subunits in vitro. PMID- 6286294 TI - Participation of the lower brain stem in induction of preovulatory gonadotropin surges in female rats. AB - The role of the lower brain stem in controlling preovulatory gonadotropin surges was investigated in female rats under acute experimental conditions. Electrolytic lesions or diethyldithiocarbamate implantations in the ventrolateral part of the medulla oblongata (VLMO), which were carried out at 1100-1330 h on the day of proestrus, resulted in a blockade of the preovulatory surges of LH, FSH and PRL as well as subsequent ovulation. Such treatments in the dorsomedial part of the medulla oblongata did not affect gonadotropin surges or ovulation. By means of electrolytic lesions in the VLMO, norepinephrine concentrations were significantly reduced in the preoptic-anterior hypothalamic area at 1700-1800 h on proestrus, though they did not change in the mid-posterior hypothalamus. Electrochemical stimulations of the suprachiasmatic part of the preoptic area or norepinephrine injections into the third ventricle at 1400-1500 h on proestrus in animals with VLMO lesions succeeded in induce gonadotropin surges and ovulation. These results suggest that the lower brain stem is involved in the induction of preovulatory gonadotropin surges and that the process may be mediated by the ascending noradrenergic system which originates in the VLMO. PMID- 6286295 TI - Improvement in abnormal secretion of thyrotropin and gonadotropin after restoration of serum calcium is pseudohypoparathyroidism. AB - A 25 yr-old woman patient was admitted because of convulsion. The diagnosis of pseudohypoparathyroidism was made on the basis of typical stigmata, lowered serum calcium, increased serum phosphorus and parathyroid hormone levels, and defective response in urinary excretion of cyclic AMP and phosphorus to exogenous parathyroid extract. Endocrine studies performed in the hypocalcemic state revealed several abnormalities of the pituitary gland such as an exaggerated response of TSH to TRH, high basal levels and exaggerated responses of LH/FSH and a blunted GH response to arginine-HCl, while there was no clinical evidence of hypothyroidism and hypogonadism. These abnormalities of anterior pituitary function were normalized after the restoration of normocalcemia by using 1 alpha hydroxy-cholecalciferol. These results suggest that some endocrine abnormalities observed in pseudohypoparathyroidism might be functional and reversible disorders secondary to hypocalcemia rather than genetic ones. PMID- 6286296 TI - Various phenotypes of diabetes mellitus at ultimate outcome of acutely developed diabetic state induced by viral infection. AB - For 4 to 8 years we followed up 3 diabetic patients in whom the onset of diabetes seemed to be closely related to the well-documented Epstein-Barr virus infection (Case 1) or Coxsackie B4 virus infection (Case 2, 3). Although all developed acute ketosis-prone diabetes in the convalescent stage of the viral infections, the subsequent clinical courses were quite different from each other. Case 1 has remained consistently insulin-dependent and associated with positive islet cell antibody, gastric parietal cell antibody, thyroglobulin hemoagglutinating antibody and thyroidal microsomal hemoagglutinating antibody. Case 2 restored normal glucose tolerance. Case 3 has become noninsulin-dependent diabetes mellitus after a 6 year interval. Thus, it is reasonably presumed that virus could be responsible for the occurrence of different phenotypes of diabetes. PMID- 6286297 TI - Effect of drug treatment and aerosoled antigen sensitization on cyclic AMP and beta adrenergic receptors of guinea pig lung. AB - The interaction between the number of beta adrenergic binding sites and the ability of a beta adrenergic agonist, isoproterenol, to increase cyclic AMP content of guinea pig lung slices was studied. A complex relationship was found. Chronic sensitization of the guinea pig to an aerosol of ovalbumin resulted in lung slices which were hyporesponsive to isoproterenol in vitro, yet possessed an unchanged number of beta adrenergic binding sites. Chronic exposure of guinea pigs to aerosoled isoproterenol or acute treatment with hydrocortisone did not change the number of beta adrenergic binding sites or the responsiveness of the tissue to isoproterenol in vitro. However, chronic hydrocortisone treatment increased the number of binding sites found on the lung slices by 44%, yet there was no change in the responsiveness of the tissue to isoproterenol in vitro. These data suggest that drugs and disease may change the relationship between the various components of the beta adrenergic binding-adenylate cyclase complex of lung. PMID- 6286298 TI - Iprindole-induced phospholipidosis in rat alveolar macrophages: alterations in oxygen consumption and release of oxidants. AB - Rats chronically treated with the cationic amphipilic drug iprindole developed a phospholipid storage disorder in their pulmonary alveolar macrophages (AMs). AMs from these iprindole-treated rats (IP-AMs) were compared to AMs from control rats (C-AMs) regarding oxygen consumption and the release of two reactive oxygen species, superoxide anion and hydrogen peroxide. Responses of C-AMs and IP-AMs were compared at rest and when stimulated by unopsonized or opsonized zymosan. Opsonization was not necessary in order to induce respiratory burst-associated phenomena in either cell type; in fact in all cases, for given cell type responses to unopsonized zymosan were virtually identical to those of opsonized zymosan. When at rest, IP-AMs consumed oxygen at a rate nearly identical to that of C-AMs. When stimulated with zymosan particles, IP-AMs consumed more oxygen than controls. However, when superoxide anion and hydrogen peroxide, two products of the respiratory burst were measured, IP-AMs released less of these species than C-AMs when at rest and when particle stimulated. Despite the lower total release of these species by the IP-AMs, the zymosan-induced release (stimulated minus resting levels) was greater for these cells than C-AMs. Therefore, the IP AMs were found to be more responsive to the zymosan particle than C-AMs. The results indicate marked changes in the release of reactive oxygen species from the AMs following induction of phospholipidosis. PMID- 6286299 TI - The effect of oxygen-derived free radicals on pulmonary endothelial cell function in the isolated perfused rat lung. AB - Oxygen-derived free radicals have been shown to damage endothelial cells in tissue culture. We have extended these observations to demonstrate a direct toxic action of oxy radicals on endothelial cell function in the isolated perfused rat lung model. Using an enzymatic system of hypoxanthine and xanthine oxidase, we generated a superoxide flux of 5-7 nmol/min within the pulmonary circulation. This resulted in alteration of endothelial cell function, including a progressive fall in the cellular transport of (14C) 5-hydroxytryptamine, edema formation manifested by a falling dry weight ratio and increasing mean transit times appearance of thiobarbituric acid reactive material in the perfusate indicating lipid peroxidation, and histologic evidence of edema formation with disruption of the endothelial cell plasma membrane. Inhibitor studies confirmed that the observed effects were due to superoxide radical generation. Oxygen derived free radicals may play a role in the pulmonary endothelial cell damages seen in inflammatory disorders and oxidant lung injury. PMID- 6286300 TI - The sensitivities of creatine and adenylate kinases to the neurotoxins acrylamide and methyl n-butyl ketone. PMID- 6286301 TI - DNA fragment conformations. 1H-NMR studies of helix-coil transition, conformations and dynamic structures of the self-complementary deoxyhexanucleotide d(A-C-A-T-G-T) in aqueous solution. PMID- 6286302 TI - Base-excision repair in carrot cells. Partial purification and characterization of uracil-DNA glycosylase and apurinic/apyrimidinic endodeoxyribonuclease. AB - Uracil-DNA glycosylase and apurinic/apyrimidinic (AP) endodeoxyribonuclease have been purified from cultured carrot cells. The two enzymes, separated by affinity chromatography on Sepharose-poly(rU), were found to have properties similar to those of the homologous bacterial and mammalian enzymes. The action of AP endodeoxyribonuclease on (dA)230 . (dT, dU)230 partially depyrimidinated by uracil-DNA glycosylase suggests that these two enzymes might act successively to initiate the repair of uracil-containing DNA. PMID- 6286303 TI - Substrate specificity studies on a calf thymus nuclear phosphoprotein kinase. AB - 1. The protein kinase activity associated with the phosphoprotein fraction of calf thymus nuclei has been examined for its ability to phosphorylate exogenous phosphoprotein substrates. beta-Casein and phosvitin are both efficient phosphate acceptors. Phosphorylation of alpha s1 and beta-caseins was shown, directly, to occur at threonyl-49 and threonyl-41 respectively. Both threonyl residues occur within the sequence Thr-Glu-Asp. 2. alpha s1-Casein becomes a much better substrate for the kinase when partially dephosphorylated. A similar response is shown by a phosphopeptide containing the alpha s1-casein phosphate cluster and is due to a new phosphorylation site becoming available. Efficient phosphorylation of beta-casein requires that the phosphate cluster (residues 15-19) be intact and results are presented which are consistent with there being a similar requirement for phosphorylation of the site created in alpha s1-casein by partial dephosphorylation. 3. Comparison of genetic variants of beta-casein as phosphate acceptors for the kinase shows that the presence of lysyl residues close to the phosphorylation site markedly depresses the rate of phosphorylation. Maleylation of beta-casein increases the rate of phosphorylation for all of the variants tested, although to varying extents. Treatment of maleylated beta-casein with trypsin to remove the N-terminal phosphopeptide inhibits phosphorylation by the kinase. 4. The structural determinants of beta-casein allowing it to be efficiently phosphorylated by the kinase are concluded to be the presence of a sequence surrounding the phosphorylation site, which is rich in acidic amino acid residues and from which basic residues are absent. The acidic phosphate cluster of beta-casein also promotes phosphorylation either by interacting directly with the enzyme or through its influence on the conformation of beta-casein. PMID- 6286304 TI - Partial P1 nuclease digestion as a probe of tRNA structure. AB - Partial P1 nuclease digestion of 5'-32P-labeled tRNAs, followed by polyacrylamide gel electrophoresis and autoradiography, results in a series of bands of unequal intensities. Comparison of the digestion profiles of two conformers of yeast tRNA3Leu shows that P1 nuclease is sensitive to structural features of its substrate. Examination of the digestion pattern of yeast tRNAPhe in light of its known three-dimensional conformation shows that the enzyme's activity reflects the primary, secondary and tertiary levels of tRNA structure. PMID- 6286305 TI - Macromolecular complex of aminoacyl-tRNA synthetases from sheep liver. Identification of the methionyl-tRNA synthetase component by affinity labeling. AB - Both the tRNA aminoacylation and amino-acid-dependent ATP-PPi exchange activities of monomeric trypsin-modified methionyl-tRNA synthetase from sheep liver are lost upon incubation with oxidized initiator tRNAMet. The inactivation, which reflects the formation of a Schiff's base between the 5'-terminal adenosine of tRNA and a lysine within the catalytic site of the enzyme, is accompanied by the covalent attachment of one tRNA molecule per enzyme molecule. The affinity labeling method is applied to the sheep liver complex of Mr 10(6) carrying seven aminoacyl-tRNA synthetase activities, from which the monomeric trypsin-modified methionyl-tRNA synthetase (Mr 68 000) was derived. Upon incubation with oxidized initiator tRNAMet, the methionyl-tRNA synthetase activity of the complex is lost. Of the eleven polypeptide chains composing the high-molecular-weight complex, only one polypeptide chain with Mr 103 000 reacts with the modified tRNAMet. The blocking by periodate-treated tRNA of the methionyl-tRNA synthetase activity in the complex has no effect on the other aminoacyl-tRNA synthetase activities. This strongly argues in favor of the independent parallel functioning of the seven aminoacyl-tRNA synthetases associated in a high-molecular-weight complex. PMID- 6286306 TI - Maturation of exported proteins in Escherichia coli. Requirement for energy, site and kinetics of processing. AB - Precursor forms of periplasmic and outer membrane proteins were accumulated in phenethyl-alcohol-treated cells in membrane fractions. After removal of phenethyl alcohol, maturation occurred in the absence but not in the presence of carbonylcyanide m-chlorophenylhydrazone (30 microM). The site and kinetics of processing were investigated for OmpA, LamB and OmpF proteins and for maltose binding protein and TEM beta-lactamase. With regard to sites of processing, no fundamental difference between outer membrane and periplasmic proteins was observed. For maltose binding protein and TEM beta-lactamase, maturation, like that of outer membrane protein precursors, occurred in membrane fractions. Processing of pro-OmpA protein was about as fast as that of pro-LamB protein whereas pro-OmpF protein appeared to mature more slowly. While carbonylcyanide m chlorophenylhydrazone (30 microM) prevented processing of all precursor forms, arsenate, which alters formation of ATP even when it was used at 1 mM, did not totally prevent maturation occurring. These results are discussed with regard to the biosynthesis and assembly of exported proteins. PMID- 6286307 TI - Involvement of prostaglandin E and adenosine 3', 5'-monophosphate in lipopolysaccharide-stimulated collagenase release by rat Kupffer cells. AB - Kupffer cells exposed to bacterial lipopolysaccharide in vitro synthesized collagenase and released the major portion of it into the extracellular space while the intracellular level of enzyme was not altered significantly. Cycloheximide prevented the appearance of collagenase in the medium indicating de novo synthesis. Indomethacin, an inhibitor of cyclooxygenase, also blocked collagenase synthesis. In line with this observation. Kupffer cells were found to synthesize substantial amounts of prostaglandin E2 when exposed to lipopolysaccharide; concomitantly, cellular cAMP levels were increased. Indomethacin was shown to abolish the stimulated cAMP formation. Addition to the culture medium of cAMP or dibutyryladenosine 3', 5'-monophosphate as well as of prostaglandin E2 or, to a lesser extent, prostaglandin E1 allowed indomethacin inhibited cells to resume the production of collagenase. It is proposed that in rat Kupffer cells lipopolysaccharide-elicited collagenase synthesis and excretion is mediated sequentially by stimulated production of prostaglandin E2, enhanced adenylate cyclase activity and increased intracellular cAMP levels. PMID- 6286308 TI - Direct measurement of proton transfer as a probing reaction for the microenvironment of the apomyoglobin heme-binding site. AB - Aromatic alcohols fluoresce at different wavelengths in their neutral (phiOH*) and anionic (phiO-*) excited states. Consequently, time-resolved fluorescence measurements, at the respective wavelengths, can be used for measuring the rates of proton dissociation and recombination of the excited molecule. As the lifetime of the excited state is very short (a few nanoseconds), the measured reaction is that which takes place in a volume corresponding to the diffusion distance of the proton during the lifetime of the excited state. 8-Hydroxypyrene 1,3,6 trisulfonate (pK = 7.7, pK* = 0.5) is bound to apomyoglobin with a stoichiometry of 1:1. In the bound state its neutral form fluorescence increase 20-fold. The binding affinity is pH-dependent. Two protonatable groups, with pK = 6.5, participate in the stabilization of the negatively charged ligand in the binding site. The ligand is bound only to the apoprotein and is displaced from its site by hemin. Thus we suggest that the ligand is bound to the heme binding site of apomyoglobin. Time-resolved fluorescence of the bound ligand yields the rate constants of proton dissociation and recombination as taking place within the heme binding cavity of apomyoglobin. The rate of proton dissociation is slowed to 7% of the rate measured for the free ligand. Such a slow dissociation indicates a strong interaction of the water in the cavity with the walls [Gutman, M., Huppert, D., and Nachliel, E. (1982) Eur. J. Biochem. 121, 637-642]. The water activity in the site is equivalent to alpha (H2O) = 0.67. PMID- 6286310 TI - Interactions of ellipticine with model or natural membranes. A spectrophotometric study. AB - Ellipticine is the type of a group of substances used against human cancer. It was shown that ellipticine and 9-methoxyellipticine strongly interact with monolayers of negatively charged phospholipids. In the present paper it is shown by spectrophotometric methods that ellipticine interacts with suspensions of a model membrane with a 1/1 charge neutralization, and with a natural membrane, both negatively charged. At a physiological pH, ellipticine undergoes protonation in the presence of these membranes. Its apparent pK is increased by 1, and its spectral behaviour, when the pH is changed, indicates that the drug is in a less polar environment, suggesting that hydrophobic bonds link the drug and the lipids. The existence of such bonds is suggested by the fact that the drug is not significantly released from anionic membranes when the pH is increased to 10, at which value ellipticine is not ionized. The consequences of these ionic and hydrophobic interactions are briefly discussed. PMID- 6286309 TI - Purification and characterization of the cytochrome c oxidase from Rhodopseudomonas sphaeroides. AB - When grown aerobically in the dark, Rhodopseudomonas sphaeroides develops a respiratory chain similar to that in mitochondria and the photosynthetic apparatus is suppressed. The aa3-type cytochrome c oxidase from Rps. sphaeroides has been purified in Triton X-100 by affinity chromatography with Sepharose 4B coupled to yeast cytochrome c. The oxidase contains 14 nmol heme a/mg protein and is composed of three polypeptide subunits with relative molecular masses of 45000, 37000 and 35000. The enzyme is highly active in the presence of detergents, with a maximal velocity of 300 s-1/mol oxidase using either yeast or horse-heart cytochrome c. The Rps. sphaeroides oxidase is cross-reactive with antibodies directed against the oxidases from Paracoccus denitrificans and Saccharomyces cerevisiae. A particularly close relationship is indicated in the case of P. denitrificans. The Rps. sphaeroides oxidase has been incorporated into phospholipid vesicles. The resulting oxidase in these vesicles demonstrates high enzymatic activity and a respiratory control ratio of 5. Using these vesicles, no evidence for proton extrusion accompanying cytochrome c oxidation was observed. The data suggest that the Rps. sphaeroides oxidase does not function as a proton pump. PMID- 6286311 TI - The reaction of bovine glutamate dehydrogenase with periodate-oxidised ADP. AB - 1. The reactive analogue oADP produced by periodate oxidation of ADP has been studied as a potential affinity label for the enzyme bovine glutamate dehydrogenase, using circular dichroism (CD) difference spectroscopy to monitor specific binding. 2. The analogue binds stoichiometrically, rapidly and reversibly to the adenine nucleotide binding site with Kd approximately equal to 12 microM (20 degrees C, pH 7) with characteristic intensification of the adenine nucleotide CD at 260 nm. 3. This complex is unstable and decays with a half-life of about 1.5 h; the analogue then becomes attached as a Schiff base to a number of subsidiary sites, including the enzyme active site, with partial inactivation of the enzyme. 4. Depending upon initial concentration of oADP, the enzyme activity is progressively lost during the slow reaction; following borohydride reduction, up to four molecules of analogue are bound/subunit. 5. Protection against loss of enzyme activity is afforded by the coenzyme NAD+ plus glutarate or L-hydroxyglutarate (an effective inhibitor), or by glutarate alone, but not by NAD+ alone. 6. Spectroscopic and protection studies indicate that after the decay of the specific CD signal, the enzyme retains the capacity to bind ADP, but that this is progressively lost in parallel with decay of enzymic activity. 7. The results are consistent with proximity or functional interaction between the adenine nucleotide site and the coenzyme binding portion of the active site. 8. Thus oADP does not act as a true affinity label for the adenine nucleotide binding site, but the reaction subsequent to binding at that site shows some specificity directed towards the active site. PMID- 6286312 TI - Kinetic and structural differences between cytochrome c oxidases from beef liver and heart. AB - 1. The cytochrome content of beef liver mitochondria differs from that of beef heart mitochondria by an eightfold lower cytochrome aa3 and a twofold lower cytochrome b and c + c1 content. 2. The kinetic properties of cytochrome c oxidases from beef liver and heart were measured with intact cytochrome c depleted membranes, deoxycholate-dissolved membranes, and with the isolated enzymes at various cytochrome c concentrations with an oxygen electrode. Under all conditions a higher V was found for the liver enzyme, both for the low affinity and for the high-affinity binding site for cytochrome c. Differences were also found for the Km of the two enzymes. 3. Isolated beef heart mitochondria contained about twice as much cardiolipin than beef liver mitochondria. The isolated enzymes contained one mole cardiolipin per mole of the heart enzyme, but 2 moles cardiolipin per mole of the liver enzyme. 4. By application of a high performance sodium dodecylsulfate gel electrophoretic system the two isolated enzymes could be separated into 13 different protein components, three of which (polypeptides VIa, VIIa and VIII) were found to differ in their apparent molecular weights. The functional meaning of cytochrome c oxidase isoenzymes in liver and heart is discussed. PMID- 6286313 TI - Induction of ornithine decarboxylase in guinea pig lymphocytes by the divalent cation ionophore A 23187. Effect of dibutyryladenosine 3',5'-monophosphate. AB - The divalent cation ionophore, A23187, at a concentration of 0.25 microgram/ml, enhanced influx of Ca2+, activity of ornithine decarboxylase and incorporation of [3H]thymidine into DNA of guinea pig lymphocytes. Combined treatment of cells with A23187 and dibutyryladenosine 3',5'-monophosphate (Bt2cAMP) augmented these three events. A23187 at a concentration of 0.06 microgram/ml was insufficient for induction of ornithine decarboxylase stimulated neither Ca2+ influx nor [3H]thymidine incorporation, but stimulated Ca2+ efflux. A23187 (0.06 microgram/ml) in combination with Bt2cAMP caused a marked induction of ornithine decarboxylase and stimulation of [3H]thymidine incorporation into DNA. When the time of Bt2cAMP addition was delayed after A23187, the stimulation of ornithine decarboxylase activity decreased. Washout of Bt2cAMP from cell culture earlier than 4 h of incubation caused a reduction in the stimulatory effect of Bt2cAMP. These results suggest that raising concentrations of cytoplasmic Ca2+ and cellular cAMP are important to some initial events leading to induction of ornithine decarboxylase and these biochemical changes are obligatory sequential steps for stimulation of DNA synthesis. PMID- 6286314 TI - Grey-scale ultrasonography in the evaluation of primary carcinomas of the bile ducts. AB - The authors report the echographic findings observed in 24 proven cases of primary carcinoma of the bile ducts. Most tumours, either vegetating or infiltrating with extraductile extension, were recognized on ultrasound. This technique turned out to be accurate in detecting peripheral, hilar and distal ductal carcinomas. Ultrasonography complements PTC and ERCP. Their combined use allows the surgeon a better selection of the cases and of the proper operative procedure. PMID- 6286315 TI - Haemodynamics and myocardial metabolism of phosphorus depleted dogs: effects of catecholamines and angiotensin II. AB - The responses of arterial pressure and myocardial contractile force (VPM) to infusion of angiotensin II, noradrenaline and orciprenaline were examined in twelve dogs during a control phase, after 30 days of dietary phosphorus deprivation and after 21 days of phosphorus repletion. In the phosphorus depletion period, animals had low skeletal and heart muscle Pi content, low magnesium, ATP and creatine phosphate in skeletal and heart muscle with no change of ADP, AMP or energy charge. In the basal state, VPM was diminished with no change of end-diastolic and systolic pressure. Infusion of angiotensin II caused a significantly smaller rise of arterial pressure (angiotensin II resistance), and the stimulatory effect of noradrenaline and orciprenaline on VPM was diminished (catecholamine resistance). These effects were reversible with Pi repletion. In phosphorus depletion, arterial concentrations were increased for lactate, unchanged for FFA and decreased for acetoacetate/beta-hydroxybutyrate. Unchanged myocardial extraction of lactate or beta-hydroxybutyrate and preserved cell Pi uptake for glycogenolysis were observed. The initial rate of uptake of calcium and concentrating ability of myocardial sarcoplasmic reticulum were unchanged. PMID- 6286316 TI - The effect of chlormethiazole on plasma adrenocorticotrophic hormone concentrations in chronic alcoholics. AB - Plasma adrenocorticotrophic hormone (A.C.T.H.) levels were estimated by radioimmunoassay in 20 male chronic alcoholics (mean age +/- SD - 40.55 +/- 7.83) both before and during treatment with chlormethiazole. There was no statistically significant difference between the pre-treatment (mean +/- SD =25.25 +/- 14.87 ng/1) and post-treatment levels (mean +/- SD = 26.45 + 15.32) of plasma A.C.T.H. (paired "t" = 1.83; N.S; log transform "t" = 2.79; N.S; p less than 0.05). So, chlormethiazole has not been found to influence plasma A.C.T.H. levels adversely but because of its sedative effect, it may eliminate the ethanol-induced stress thereby helping indirectly in reducing the elevated levels. Plasma A.C.T.H. levels seem to be raised in some patients due to chronic ethanol ingestion and total abstinence for more than a week might be needed for the raised levels to return towards normal diurnal pattern. PMID- 6286317 TI - Effect of urinary pH on the disposition of methadone in man. AB - The influence of urinary pH on the acute disposition of methadone in man was studied in five healthy volunteers. A cross-over experiment was performed in each subject. In the first experiment the subjects were treated with ammonium chloride (urinary pH approximately 5.2) and in the other the urine was made alkaline (pH approximately 7.8) by treatment with sodium hydrogen carbonate. d, l-Methadone HCl 10 mg (M) was administered intramuscularly on each occasion and blood, saliva and urine levels of M were determined by mass fragmentography. Plasma half-lives, volumes of distribution and body clearances of M were calculated in both experiments. The plasma half-lives in the beta-phase were 19.5 +/- 3.6 h (acidic urine) and 42.1 +/- 8.8h (alkaline urine), respectively (p less than 0.001). The volumes of distribution were increased when the pretreatment was changed from ammonium chloride to sodium bicarbonate, namely from 3.51 +/- 0.41 l/kg to 5.24 +/- 0.83 l/kg (p less than 0.01). The body clearance decreased from 134 +/- 21 ml/min (acidic) to 91.9 +/- 9.1 ml/min (alkaline urine) (p less than 0.01). The ration M plasma/M RBC was about 2.3 and the elimination of M from RBCs was good agreement with the plasma kinetics of M under both experimental conditions. The salivary levels of M did not reflect the plasma kinetics and considerable variation was seen in the ratio M saliva/ M plasma (0.26-2.98). Thus, the present experiments demonstrate that pretreatment either with ammonium chloride or bicarbonate had profound effects on both the distribution and elimination kinetics of methadone. PMID- 6286318 TI - Cerebellar cyclic GMP and behavioral effects after acute and repeated administration of benzodiazepines in mice. AB - The previous finding that benzodiazepine-induced changes in cyclic GMP in the cerebellum were poorly correlated with the impairment of motor function has now been extended further. The effects following acute, and during repeated administration of diazepam, quazepam and flurazepam were studied in mice. A 10 day period of daily treatment with 5 mg/kg p.o. was used in these studies. The content of cGMP was significantly reduced (P less than 0.05) during the chronic administration of quazepam and flurazepam except after 10 days for flurazepam. After diazepam, cGMP levels were lower than in control animals but a significant difference was observed only at the sixth and tenth day because a minimal threshold dose was used. Antagonism of pentylenetetrazol-induced convulsions was significantly (P less than 0.05) effective over the 10 day treatment for all drugs. Spontaneous motor activity was reduced by quazepam given for two days and by diazepam given for one day while this measure was unaffected by flurazepam given at the same dose which was effective in the other tests. These results further supported a lack of association between changes of cGMP content in the cerebellum and sedative or muscle relaxant effects of benzodiazepines, whereas the lowering of cGMP levels seemed to parallel effects that are not subject to tolerance such as the antagonism of pentylenetetrazol-induced convulsions. PMID- 6286319 TI - The action of barbiturates on contractile responses of canine and feline bronchial smooth muscle. AB - Pentobarbitone, thiopentone and phenobarbitone in concentrations ranging from 10( 4) M to 10(-3) M reduced the contractile responses of isolated canine and feline trachealis muscle to electrical field stimulation without affecting contractile responses to acetylcholine. The order of potency was thiopentone, pentobarbitone and phenobarbitone. Experiments using carbachol and neostigmine or physostigmine suggested that the barbiturates act by preventing the release of acetylcholine from nerve endings. In vivo, pentobarbitone (2.5-20 mg/kg i.v.) produced dose related reductions in the increase in total lung resistance induced by vagal stimulation in chloralose-anaesthetised cats. PMID- 6286320 TI - Ethyl beta-carboline-3-carboxylate reverses the diazepam effect on cerebellar cyclic GMP. AB - The administration of ethyl beta-carboline-3-carboxylate (beta-CCE), a ligand for benzodiazepine receptors, did not affect the cerebellar cyclic GMP level in mice, but giving beta-CCE together with diazepam significantly inhibited the diazepam induced decrease in cyclic GMP. The fact that no antagonism was observed when beta-CCE was given 15 min before the diazepam treatment indicates that beta-CCE is short-acting. These biochemical observations support the conclusions from behavioral and electrophysiological studies which indicated that beta-CCE is a short-acting antagonist of benzodiazepines. PMID- 6286322 TI - Measurement of alpha-adrenoceptor "turnover' using phenoxybenzamine. PMID- 6286321 TI - Prostaglandins A as possible endogenous ligands of benzodiazepine receptor. AB - We carried out biochemical studies on several prostaglandins to determine if whey would displace [3H]diazepam binding to the membranes of bovine cerebral cortex. Prostaglandins A1 and A2 were the most potent inhibitors of this binding and all the others tested were either less potent or not effective. As prostaglandins A1 and A2 competitively inhibited [3H]diazepam binding with Ki values of 7.1 +/- 0.1 and 15 +/- 1 microM, respectively, their possible function as endogenous ligands of benzodiazepine receptors warrants further attention. PMID- 6286323 TI - Classification of some GABA antagonists with regard to site of action and potency in slices of rat cuneate nucleus. AB - Compounds reported to be GABA antagonists have been studied quantitatively on dorsal funiculus fibres and terminals in the rat cuneate nucleus in vitro. The potencies of the antagonists against the GABA analogue muscimol were determined as pA2 values. Distinction was made between three different sites of antagonist action within the GABA receptor and ionophore complex. Competitive antagonists, presumed to act at the GABA receptor, and their pA2 values were bicuculline (5.98), bicuculline methochloride (5.88), strychnine (5.29) and tubocurarine (4.95). Antagonists which were not competitive and acted predominantly at the 'picrotoxin site' on the ionophore were picrotoxin (6.19), picrotoxinin (6.03), isopropylbicyclophosphate (5.82) and leptazol (2.89). A third type of antagonism was shown by frusemide. Attention is drawn to the picrotoxin site and its likely importance in the regulation of GABA-mediated inhibition by drugs. PMID- 6286324 TI - Chlorpromazine inhibition of enterotoxin-induced fluid secretion and cAMP production in rat ileum. AB - A heat-labile enterotoxin prepared from E. coli (EcLT) increased fluid secretion and cAMP production by segments of rat ileum in vivo and in vitro. The effect of this toxin was compared to that of cholera toxin (VcLT). The increase of cAMP occurred more rapidly after EcLT than after VcLT indicating a difference in the kinetics of uptake or action of the two toxins. Chlorpromazine (CPZ) 5 mg/kg given by intramuscular injection 1 h before application of the toxins inhibited the increase in cAMP levels and the increase in fluid secretion in vivo. CPZ 10( 4) M given together with the toxins to intestinal loops in vitro inhibited the increase in cAMP levels and fluid secretion by this preparation. Scanning electron microscopy revealed that CPZ caused extensive shedding of the fluid producing mucosal cells. PMID- 6286325 TI - Pharmacological profiles of beta-funaltrexamine (beta-FNA) and beta chlornaltrexamine (beta-CNA) on the mouse vas deferens preparation. AB - The profiles of action of beta-funaltrexamine (beta-FNA) and beta chlornaltrexamine (beta-CNA) have been assessed in the mouse vas deferens preparation. beta-FNA, but not beta-CNA, demonstrated a reversible agonist action that appeared to be mediated via kappa-receptor interaction. beta-CNA produced an irreversible antagonism of mu-, kappa- and delta-mediated agonist actions, whereas beta-FNA irreversibly antagonized mu-mediated agonist effects only. This selective action of beta-FNA could also be seen following administration in vivo. beta-CNA and particularly beta-FNA should prove valuable in the elucidation of multiple opioid receptors. PMID- 6286326 TI - Effects of ethidium bromide on the nicotinic acetylcholine receptor. AB - Ethidium bromide was tested for electrophysiological effects at the postsynaptic membrane of the frog muscle endplate. At low concentrations ethidium bromide blocked the open ion channel of the nicotinic acetylcholine receptor and reduced its open time. The rates for channel blocking and unblocking were calculated giving a dissociation constant of 139 nM at -80 mV membrane potential. PMID- 6286327 TI - Caerulein and cholecystokinin suppress beta-endorphin-induced analgesia in the rat. AB - The analgesic action of beta-endorphin, as observed in the hot plate test with rats, was effectively suppressed by intracerebroventricular (i.c.v.) injection of caerulein and cholecystokinin octapeptide (CCK-8). The effect of caerulein was particularly striking; this peptide in doses of more than 0.09 nM lessened or abolished the analgesic effect of beta-endorphin in a dose of 0.7 nM. On the other hand, non sulfated CCK-8 had no significant effect on beta-endorphin induced analgesia. PMID- 6286328 TI - The effects of receptor selective opioid peptides on morphine-induced analgesia. AB - Utilizing the mouse tail-flick assay, four opioid peptides, which have been reported to be selective for either mu- or delta-opioid receptors, were examined for their analgesic potency and for their ability to modify morphine-induced analgesia. [D-Ala2,D-Leu5]enkephalin and [D-Ser2,Thr6]leucine-enkephalin, putative delta-receptor selective peptides, produced a potent analgesic response and at subanalgesic doses potentiated morphine-induced analgesia. Morphiceptin and [D-Ala2,Pro5]enkephalinamide, putative mu-receptor selective peptides, were similarly found to produce analgesia. However, in contrast to the delta-receptor selective peptides, three mu-receptor selective peptides were unable to alter the potency of morphine. Thus, it would appear that the potentiation of morphine analgesia is a unique property of delta-receptor selective peptides. PMID- 6286329 TI - Inactivation of a basement membrane component responsible for cell proliferation but not for cell attachment. PMID- 6286330 TI - Morphological changes in Ehrlich ascites tumor cells during the cell fusion reaction with HVJ (Sendai virus). III. Morphological characterization of HVJ glycoproteins integrated into the plasma membrane and their internalization by coated vesicles. PMID- 6286331 TI - Endotoxin-induced degranulation of the Limulus amebocyte. PMID- 6286332 TI - Improved system for capillary microinjection into living cells. PMID- 6286333 TI - Early initiation of bovine satellite I DNA replication. PMID- 6286334 TI - Expansion of chicken erythrocyte nuclei upon limited micrococcal nuclease digestion. Correlation with higher order chromatin structure. PMID- 6286335 TI - The influence of aging on intestinal absorption of vitamins A and D by the rat. AB - The absorption of vitamins A and D was examined in 6, 12, and 24 months old female Wistar rats. Animals were dosed by stomach tube with radioactive forms of vitamins A and D and killed 18 hours later. Isotope in feces collected during this time and isotope remaining in the gastrointestinal tract were determined and subtracted from total dose to determine percentage of absorption. Under the conditions of this study, age had no significant effect on the absorption of vitamins A and D. PMID- 6286336 TI - Altered activation of rat hepatic cyclic AMP-dependent protein kinases with increasing age. AB - The activity ratio of hepatic cyclic AMP-dependent protein kinase has been found to change with increasing age in male, Long-Evans rats. In rats 35 days old (135 g), the activity ratio (cyclic AMP-dependent protein kinase activity measured in the absence and presence of exogenous cyclic AMP) was 0.76, it increased to 0.92 in rats 75 days old (315 g) and to 0.98 in rats 305 days old (646 g). This change in the activity ratio seems to correlate with the ability of aging animals to respond to a stimulus, and may be a primary mechanism in the biochemistry of aging. PMID- 6286338 TI - Mianserin reduces plasma levels of beta-endorphin immunoreactivity in depressed patients. PMID- 6286337 TI - Cholinergic neurons in the basal forebrain of the cat have direct projections to the sensorimotor cortex. PMID- 6286340 TI - Effect of cuprizone feeding on hepatic superoxide dismutase and cytochrome oxidase activities in mice. PMID- 6286339 TI - AMP deaminase, 5'-nucleotidase and adenosine deaminase in rat myocardial tissue in myocardial infarction and hypothermia. AB - AMP deaminase, 5'-nucleotidase and adenosine deaminase have been estimated in skeletal muscle and myocardial tissue in normal rats and in rats subjected to experimental myocardial infarction or hypothermia. A difference in the enzyme distribution was found between the right and left ventricles in the normal rat. A decrease in the activity of 5'-nucleotidase and an increase in the activity of adenosine deaminase were observed in infarcted myocardial tissue. The activity of all 3 enzymes was found to be depressed in the myocardium in rats subjected to hypothermia. These results are discussed in relation to adenosine production and its beneficial effects. PMID- 6286343 TI - Mammalian DNA-repair endonuclease acts only on supercoiled DNA. PMID- 6286341 TI - Effects of polycyclic hydrocarbons on the induction of chromosomal aberrations in absence of an exogenous metabolic activation system in cultured hamster cells. PMID- 6286342 TI - Effects of zinc and lithium ions on the strengthening cell adhesion in sea urchin blastulae. PMID- 6286344 TI - ATP-synthetase complex (F1F0) from Escherichia coli. Purification and characterization of subunits A and B of the F0 part. PMID- 6286345 TI - Superoxide-dependent formation of hydroxyl radicals from NADH and NADPH in the presence of iron salts. PMID- 6286346 TI - Adipocytes do not falsify the superoxide theory of oxygen toxicity. PMID- 6286347 TI - Formation of conjugates by 125I-labelled ubiquitin microinjected into cultured hepatoma cells. PMID- 6286348 TI - Purine nucleoside production in mature and immature human lymphocytes. PMID- 6286349 TI - Induction of cyclic AMP-dependent phosphorylation in cytosol of differentiating mouse erythroleukemia cell line 179. PMID- 6286350 TI - Demonstration of a receptor-like binding protein for 1,25-(OH)2-D3 in cultured intestinal epithelial cells from the adult rat. PMID- 6286351 TI - Adenylate cyclase activity in permeabilised yeast. PMID- 6286352 TI - Turtle hemoglobin: evidence of a T-state oxyhemoglobin. PMID- 6286354 TI - Bilayer thickness and enzymatic activity in the mitochondrial cytochrome c oxidase and ATPase complex. PMID- 6286353 TI - Accessibility of glucose 6-phosphate: phosphohydrolase to antibody attack in modified microsomal vesicles. PMID- 6286355 TI - Oxidative inactivation of Escherichia coli by hypochlorous acid. Rates and differentiation of respiratory from other reaction sites. PMID- 6286356 TI - The Mg2+-activated phosphatidylinositol 4,5-bisphosphate-specific phosphomonoesterase of erythrocyte membrane. PMID- 6286357 TI - Parallel electrostatic effects in the interactions of superoxide with cytochrome c and with superoxide dismutase. PMID- 6286358 TI - Dissociation of CNPase dimer into catalytically active monomers by 1,4 dithiothreitol and urea. Protein-promoted reassembly. PMID- 6286359 TI - Effects of novel leukotrienes on neutrophil migration. PMID- 6286360 TI - Role of hormones and protein phosphorylation in metabolic regulation: introduction. PMID- 6286361 TI - Hormonal regulation of skeletal muscle glycogen synthase through covalent phosphorylation. AB - Studies have been initiated to determine the hormonal regulation of glycogen synthase in rabbit skeletal muscle. It was found that glycogen synthase purified from control animals was quite highly phosphorylated (2.35 mol phosphate/mol synthase subunit) with 40% of the phosphate in the trypsin-sensitive or COOH terminal domain, and 60% in the trypsin-insensitive or NH2-terminal domain. The phosphorylation state of synthase was elevated (3.9 mol/mol) by epinephrine injection and in the diabetic condition. With epinephrine, about 76% of the additional phosphate was incorporated in the trypsin-sensitive domain, which strongly supports the contention that this hormone acts through the cyclic AMP (cAMP)-dependent protein kinase. In the synthase purified from diabetic rabbits, 90% of the additional phosphate was in the trypsin-insensitive domain. Insulin treatment of the diabetics resulted in specific dephosphorylation of the trypsin insensitive domain. These results indicate that in this system insulin is not acting by inhibition of the cAMP-dependent protein kinase. PMID- 6286362 TI - Regulation by glucagon of hepatic pyruvate kinase, 6-phosphofructo 1-kinase, and fructose-1,6-bisphosphatase. AB - Glucagon stimulates gluconeogenesis in part by decreasing the rate of phosphoenolpyruvate disposal by pyruvate kinase. Glucagon, via cyclic AMP (cAMP) and the cAMP-dependent protein kinase, enhances phosphorylation of pyruvate kinase, phosphofructokinase, and fructose-1,6-bisphosphatase. Phosphorylation of pyruvate kinase results in enzyme inhibition and decreased recycling of phosphoenolpyruvate to pyruvate and enhanced glucose synthesis. Although phosphorylation of 6-phosphofructo 1-kinase and fructose-1,6-bisphosphatase is catalyzed in vitro by the cAMP-dependent protein kinase, the role of phosphorylation in regulating the activity of and flux through these enzymes in intact cells is uncertain. Glucagon regulation of these two enzyme activities is brought about primarily by changes in the level of a novel sugar diphosphate, fructose 2,6-bisphosphate. This compound is an activator of phosphofructokinase and an inhibitor of fructose-1,6-bisphosphatase; it also potentiates the effect of AMP on both enzymes. Glucagon addition to isolated liver systems results in a greater than 90% decrease in the level of this compound. This effect explains in large part the effect of glucagon to enhance flux through fructose-1,6 bisphosphatase and to suppress flux through phosphofructokinase. The discovery of fructose 2,6-bisphosphate has greatly furthered our understanding of regulation at the fructose 6-phosphate/fructose 1,6-bisphosphate substrate cycle. PMID- 6286364 TI - Transposon Tn10: genetic organization, regulation, and insertion specificity. AB - Transposon Tn10 is a composite element in which two individual insertion sequence (IS)-like sequences cooperate to mediate transposition of the intervening material. The two flanking IS10 elements are not identical; IS10-right is responsible for functions required to promote transposition, and IS10-left is defective in transposition functions. We suggest that the two IS10 elements were originally identical in sequence and have subsequently diverged. IS10-right is compactly organized with structural gene(s), promoters, and sites important for transposition and (presumably) its regulation all closely linked and, in some cases, overlapping. IS10 has a single major coding region that almost certainly encodes an essential transposition function. A pair of opposing promoters flank the start of this coding region. One of these promoters is responsible for expression in vivo of transposon-encoded transposition functions. We propose that the second promoter is involved in modulation of Tn10 transposition. Genetic analysis suggests that transposon-encoded function(s) may be preferentially cis acting. Insertion of Tn10 into particular preferred target sites is due primarily to the occurrence of a particular six-base pair target DNA sequence. The properties of this sequence suggest that symmetrically disposed subunits of a single protein may be responsible for both recognition and cleavage of target DNA during insertion. PMID- 6286363 TI - Modulation of rat liver 3-hydroxy-3-methylglutaryl-CoA reductase activity by reversible phosphorylation. AB - We have previously reported that the enzymic activity of rat liver-3-hydroxy-3 methyl-glutaryl-CoA reductase (NADPH) (HMG-CoA reductase) is modulated in vitro by a phosphorylation-dephosphorylation reaction sequence. The in vitro phosphorylation of HMG-CoA reductase was further studied by utilizing purified HMG-CoA reductase and reductase kinase. Analysis of 32P-labeled HMG-CoA reductase revealed 1 mol of phosphate per subunit. Purified [32P]HMG-CoA reductase could be dephosphorylated with phosphoprotein phosphatase. To demonstrate the in vivo phosphorylation, rats were injected with 32P and hepatic HMG-CoA reductase was isolated by immunoprecipitation and also by purification of the enzyme to homogeneity. Analysis of [32P]HMG-CoA reductase by sodium dodecyl sulfate gel electrophoresis revealed a single peak of radioactivity comigrating with HMG-CoA reductase. Administration of glucagon enhances the in vivo phosphorylation of both HMG-CoA reductase and reductase kinase. In response to glucagon, HMG-CoA reductase activity is decreased whereas reductase kinase activity is increased. These results support our concept that the enzymic activity of HMG-CoA reductase is modulated by a bicyclic cascade system involving phosphorylation dephosphorylation. The enzymic activity of HMG-CoA reductase has also been shown to be modulated by cholesterol and mevalonolactone by both short-term and long term mechanisms. The effects of cholesterol and mevalonolactone are twofold. Rapid inhibition of HMG-CoA reductase activity is due to increased phosphorylation of the enzyme; the long-term effect of HMG-CoA reductase is achieved by reduction in enzyme concentration by modulation of enzyme synthesis and/or degradation. Regulation of HMG-CoA reductase by mevalonolactone is of major importance in cellular metabolism because mevalonate serves as precursor for four separate metabolic pathways, including the formation of cholesterol, ubiquinone, dolichols, and isopentenyl tRNA. PMID- 6286365 TI - Construction of yeast strains containing genetically marked transposons. AB - Haploid yeast cells contain approximately 35 Ty (transposon yeast) elements. To facilitate the study of these elements, we have constructed yeast strains in which one of these transposons carries a genetic marker. The elements that we have modified are Ty912 and Ty917, elements originally detected at the HIS4 locus in spontaneously occurring his4- mutants. The strain constructions took place in three stages: 1) cloning of the mutant HIS4 genes containing the Ty elements; 2) introduction of a HindIII restriction fragment containing the yeast URA3 gene into the cloned transposons; and 3) replacement of the chromosomal HIS4 gene with the modified genes constructed in vitro. These strains will be extremely useful in the study of Ty transposition and other Ty-promoted DNA rearrangements. PMID- 6286366 TI - Mobile dispersed repeated DNA elements in the Drosophila genome. AB - The molecular and cytogenetic organizations of 19 nonhomologous dispersed repeated sequence families were studied in 15 different laboratory strains of Drosophila melanogaster. Elements from each of the families appear to undergo transposition within the Drosophila genome, because there were striking differences in both the number and chromosomal locations of these elements between strains. A significant fraction (greater than 1%) of Drosophila DNA therefore has an unstable genomic organization. Each middle repetitive family exhibited similar variations in the chromosomal distribution of elements between the strains. Although the movements of these elements are not limited to a small number of genomic sites, there are chromosomal regions where elements from the different dispersed repeated DNA families appear to be clustered. The locations of such preferred integration sites are different in each of the D. melanogaster strains examined. PMID- 6286367 TI - Nucleotide sequence analysis of avian retroviruses: structural similarities with transposable elements. AB - Integrated retroviral genomes are flanked by direct repeats of sequences derived from the termini of the viral RNA genome. These sequences are designated long terminal repeats (LTRs). We have determined and analyzed the nucleotide sequence of the LTRs from several exogenous and endogenous avian retroviruses. These LTRs possess several structural similarities with eukaryotic and prokaryotic transposable elements: 1) inverted complementary repeats at the termini, 2) deletions of sequences adjacent to the LTR, 3) small duplications of host sequences flanking the integrated provirus, and 4) sequence homologies with transposable and other genetic elements. These observations suggest that LTRs function in the integration and perhaps transposition of retrovirus genomes. Evidence exists for the presence of a strong promoter sequence within the LTR. The retroviral LTR also contains a "Hogness box" up-stream of the capping site and a poly(A) signal. These features suggest an additional role for the LTR in the regulation of gene expression. PMID- 6286368 TI - Biochemical mechanisms of hormone receptor-effector coupling. Introduction. PMID- 6286369 TI - Thyroid hormone-carbohydrate interaction at the hepatic nuclear level. AB - We have studied the interaction between triiodothyronine (T3) and carbohydrate (CHO) in the induction of hepatic lipogenic enzymes under both in vivo and in vitro conditions. Our studies demonstrate a synergistic relationship between T3 administration and CHO feeding in the induction of these enzymes. Likewise, in states characterized by CHO deprivation such as starvation and diabetes, the response to T3 is also inhibited. Studies in the aging animal have documented a diminished response both to CHO and to T3. Our studies suggest that T3 multiplies a primary CHO-generated signal by a constant factor, and that this signal declines with age. Additional studies with primary hepatocyte cultures provide evidence that glucose is the main factor responsible for the induction of hepatic malate dehydrogenase: decarboxylating (EC 1.1.1.40) (ME). Glucose induces ME in the absence of changes in extrahepatic hormones or metabolites and in the complete absence of T3. In the cultured hepatocyte system, T3 also acts as a constant multiplier of the primary glucose-derived signal. Our results provide further support for the thesis that the primary action of T3 at the molecular level is a multiplication of other nuclear signals. The complexity of response pattern to both T3 and CHO administration, however, is illustrated by recent studies in which we have analyzed the translated products of total poly(A+) RNA extracted from livers of rats subjected to various physiological stimuli. PMID- 6286371 TI - Response of the adrenal to adrenocorticotropic hormone (ACTH) in hyperandrogenic women treated chronically with low doses of prednisone. AB - A group of 43 women of reproductive age with clinical signs of hyperandrogenism was treated with a low dose (7.5 mg daily) of prednisone. An acute dexamethasone (DEX) suppression-adrenocorticotropic hormone (ACTH) stimulation test was performed before and after a minimum of 2 months of prednisone administration. The therapy produced a significant decrease of plasma testosterone (T) levels in a majority of patients. During the treatment period DEX failed to induce further suppression, while ACTH caused a significant rise in plasma T levels. The cortisol (F) levels were suppressed during treatment, but their response to ACTH stimulation was similar to that observed prior to initiation of therapy. PMID- 6286370 TI - Human ovarian receptors to human prolactin: implications in infertility. AB - In the human, hyperprolactinemia may interfere with fertility. To find out whether or not prolactin (PRL) can act directly on human ovarian tissues, in vitro studies involving specific binding of human PRL to various human ovarian elements were carried out. Human PRL was isolated from amniotic fluid, and its intact monomeric iodinated isohormone B was prepared. Labeled PRL was incubated with plasma membranes of either granulosa or whole follicular homogenates. Relatively high specific binding sites were obtained. Saturation studies and Scatchard analysis showed a single class of binding sites with high binding affinity (Kd = 1.8 x 10(8) M) and a concentration of 7.9 x 10(-15) moles/mg protein. These results clearly demonstrate the existence of specific receptors to isoprolactin B in ovarian elements with binding capacity and concentration at least equal to that found in other targets for PRL. It is reasonable to assume that hyperactivation of these receptors in states of hyperprolactinemia constitutes the cause for the disturbances in ovarian functions that lead to infertility. PMID- 6286372 TI - In vitro processing of avian oncovirus precursor polypeptide Pr76gag by virion protein p15. AB - In vitro cleavage by p15 virion protein of the primary translation product of the avian oncovirus gag gene, the precursor polypeptide Pr76gag, was studied in kinetic experiments. Nondenatured 35S-methionine-labelled Pr76gag, which was synthetized in reticulocyte lysate programmed by genomic AMV-RNA was used as a substrate, pure native p15 (AMV) as a protease. Reaction conditions were optimal for in vitro protein synthesis. Composition of the cleavage products was estimated by immune precipitation with monospecific antisera against internal structural virion (AMV) proteins p27, p19, p15, and p12, and their size by SDS PAGE. Monospecificity of each of the antisera was assessed by adsorption with the three respective heterologous gag proteins, followed by immune precipitation of 35S-methionine-labelled proteins of the virion (AMV) lysate. The p15-mediated in vitro cleavage proceeds rapidly and specifically. In the early stages (10 min incubation with p15) the Pr76gag was absent, and an optimum amount of six cleavage intermediates with the following size, composition (shown in parentheses) and orientation (determined form the presence or absence of antigenic determinants of p19 or p15 proteins as N- or C-terminal moieties of the precursor) was found: N-terminal fragments of 66K (p19, p27, p12) and 60K (p19, p27); internal fragments of 37K (p27, p12); C-terminal fragments of 51K (p27, p12, p15), 32K (p12, p15) and 21K (p12, p15). These intermediates were converted into the following four final cleavage products, as the only polypeptides detected after prolonged (5 h) incubations: mature p27 and mature p15 proteins and 38K and 34K polypeptides containing only p19 antigenic determinants. Mature p12 an p19 proteins were not found among cleavage products. Autocatalytic cleavage of Pr76gag was not observed. The results hav allowed us to conclude that the arrangement of the gag proteins in the Pr76gag is N-p19-(p10?)-p27-p12-p15-C and that under in vitro conditions p15 recognizes three correct cleavage sites on native Pr76gag: one located at p12-p15 junction and two on both sides of the p27 moiety, as well as aberrant cleavage sites located inside p12 and possibly also p10 moieties. PMID- 6286373 TI - Characterization of a mutant of the Prague strain of Rous sarcoma virus inducing fusiform transformation of avian fibroblasts in vitro. AB - The DNA as isolated from duck fibroblasts transformed by a duck-adapted Prague strain of Rous sarcoma virus and used for transfection. Transformed recipient BLEF and DEF cultures exhibited considerable morphological variability. The virus designated daPR-RSV-C morphf was obtained from the culture with fusiform transformation and cloned. The virus retained the ability to induce fusiform transformation, even after 20 passages on chicken fibroblasts. There was a good correlation between focus forming activity of the virus and its tumorigenicity in chickens. The frequency of morphf mutation to another phenotype was less than 10( 3) in cloned virus. Foreign avian embryonic cells transformed by this virus clone had a similar morphological appearance as transformed chicken cells. The clone also retained two additional non-conditional markers - subgroup C specificity and the ability to replicate efficiently in duck cells ("duck adaptation"). Freshly obtained cloned virus was found not to contain a transformation-defective mutant. Such a mutant occurred in the second passage of the virus of DEF where the mutant was isolated. Inoculation of the td mutant into Brown Leghorn embryos gave rise to a sarcoma in one of the 36 examined chickens. However, no transforming virus was detected in the sarcoma. SDS-polyacrylamide gel electrophoresis showed that cloned daPR-RSV-C morphf contained only genomic RNA; its molecular weight 3.08 X 10(6) daltons corresponded to the molecular weight of a non-defective PR-RSV-C used as control. PMID- 6286374 TI - Cytoskeleton of rat neoplastic cells of spontaneous origin and their derivatives after supertransformation with avian sarcoma virus B77 in vitro. AB - Changes in the distribution of actin-containing cables (microfilament bundles), microtubules and fibronectin were studied by immunofluorescence in four rat neoplastic cell lines and compared with those of normal rat fibroblasts. No changes in the organization of microtubules were seen. A network of fibronectin filaments was observed only in normal fibroblasts whereas a few short fibronectin fibres were observed in cultures of all tumour cells. The actin-containing microfilament bundles in LW13K2 fibroblasts transformed spontaneously in vitro were shorter and differed in their orientation from those of normal fibroblasts. Almost no changes in action distribution in the supertransformed K2B773 cells, compared to LW13K2 cells, were observed. Absence of the actin-containing microfilament bundles, or only a few thin bundles were seen in cells of a spontaneous mammary carcinosarcoma SAM-LEW. Actin cables increased in thickness and number in the B77-supertransformed SAMB774 cells. The effect of pp60src on the organization of actin in neoplastic cells as compared to normal cells is discussed. PMID- 6286375 TI - Alternation in malignancy of rat spontaneous tumour cells after in vitro supertransformation by ASV. AB - Neoplastic cells from a spontaneous mammary carcinosarcoma (SAM-LEW) of LEW/CUB rats and spontaneously in vitro transformed Lewis rat embryo fibroblasts (LW13K2) were infected in vitro and transformed with avian sarcoma virus B77. Clones of B77 super-transformants (SAMB774, K2B773) were isolated and tested in vitro and in vivo for the rescue of infections B77 virus with positive results. These virogenic B77 supertransformants were then tested in the immunogenetic test of malignancy established in the LEW rat congenic system. The supertransformants exhibited restricted growth on RT1-different congenic recipients without killing them. It was in sharp contrast to parental tumours of spontaneous origin which were able to kill about one half of the RT1-different congenic recipients. This indicated a decrease in malignancy from grade IV (spontaneous parental) to grade III (ASV supertransformants) as expressed in the ITM. PMID- 6286376 TI - Effect of aflatoxin B1 on growth of normal and malignant rat fibroblasts in tissue culture (DNA synthesis). AB - The effect of aflatoxin B1 on rat embryo fibroblasts WREF, the transformed line of rat fibroblasts AWRF-1 derived from WREF and the XC line of rat fibroblasts transformed by Rous sarcoma virus has been compared. By determining growth inhibition of cultured cells and inhibition of colony formation WREF have been shown to have the sensitivity to AFB1 about 33 times greater than XC cells and 7 times greater than AWRF-1 cells. In accordance with these results, it has been found that the inhibition of DNA synthesis and the binding of 3H-AFB1 to the TCA insoluble cell fraction proceeds much more rapidly in WREF than in XC and AWRF-1 cells. These results suggest that WREF cells are capable of a more rapid metabolic activation of AFB1 to toxic metabolite AFB1-8, 9-oxide, probably owing to higher levels of cytochrome P-450-dependent microsomal oxidases. The resistance of cells to AFB1 fairly correlates with the ability of cells to grow in semisolid agar medium and the culture time. PMID- 6286377 TI - [A study on the mechanism of control of the luteal function in the pseudopregnant rat (author's transl)]. PMID- 6286378 TI - [Studies on the regulatory mechanism of ovarian hCG receptors and placental luteotropin in rats (author's transl)]. AB - The influences of exogenous hCG on rat (pseudopregnant and pregnant) ovarian hCG receptors and the presence of hCG-like material in the rat placenta were investigated. Ovarian hCG receptors in immature rats induced by PMS-hCG priming maintained a binding activity for about 20 days after hCG injection. The administration of hCG to pseudopregnant rats caused time and dose related changes in ovarian 125I-hCG binding percentage and the concentration of serum progesterone. The injection of hCG produced a slight decrease in the binding percentage and was followed by a marked decrease. The measurement of ovarian tissue hCG concentration by radioimmunoassay suggested that the initial loss of binding percentage was associated with the occupancy of receptors by the exogenous hCG, but the major loss of binding percentage might not be attributed to the receptor occupancy. The active process might be involved in the mechanism of receptor loss. The progesterone concentration in sera had a tendency to increase after the injection of 25IU hCG, while it decreased after the injection of 125IU hCG. The 125I-hCG binding percentage to ovaries and the serum concentration of progesterone reached their maximal values from mid to late pregnancy and declined as parturition approached. After the injection of hCG into pregnant rats, the binding percentage rapidly decreased and the serum concentration of progesterone increased. Thereafter the binding percentage recovered on the 4th day after the injection of hCG. On that day, though the concentration of hCG in ovarian tissue was barely detectable, the binding percentage was significantly low compared to the control level. The rat placenta (8th to 21st day of pregnancy) contained hCG-like activities, but they could not be detected in peripheral blood. In the treatment of hCG-antibody in pregnant rats, the binding percentage did not change but the serum concentration of progesterone showed a tendency to decrease to the control level. These studies show that hCG receptors exist as free sites in pregnant and pseudopregnant rat ovaries and may be regulated by down regulation after the treatment of large amounts of exogenous hCG. We found that hCG-like material in rat placenta determined by radioimmunoassay might serve as a luteotropin during pregnancy. PMID- 6286379 TI - [Studies on the measurement of vitamin D derivatives in human plasma. III. Plasma concentrations of vitamin D derivatives in various diseases (author's transl)]. AB - Plasma concentrations of vitamin D3, 25-OH-D3, 24, 25(OH)2D3 and 1, 25(OH)2D3 were measured in patients with various diseases using the multiple assay system previously reported. In patients with hyperparathyroidism, the plasma levels of 1, 25(OH)2D3 tended to increase, while the levels of 24, 25(OH)2D3 tended to decrease in many cases. On the other hand, plasma 1, 25(OH)2D3 levels were low, while 24, 25(OH)2D3 levels were high in patients with hypoparathyroidism. No significant differences in the levels of plasma D3 derivatives among idiopathic-, postoperative- and pseudo- hypoparathyroidism were observed. In a majority of hemodialyzed patients with advanced renal failure who showed no overt bone changes on X-ray films, both the plasma 1, 25(OH)2D3 and 24, 25(OPH)2D3 levels were distributed from normal to very low. In a majority of patients with osteoporosis and hypoparathyroidism, plasma 1, 25(OH)2D3 levels rose quickly after the administration of 1 alpha-OH-D3. In hypophosphatemic vitamin D resistant rickets, however, the response to the relatively large amounts of 1 alpha-OH-D2 was poor, although the concentrations of plasma 24, 25(OH)2D3 were elevated by the 1 alpha-OH-D3 administration in most of these cases. The plasma 1, 25(OH)2D3 and 24, 25(OH)2D3 concentrations in patients with senile osteoporosis were distributed from lower to higher range than those of normal adults. There was a relatively good correlation between plasma levels of 25-OH-D3 and D3 only in cases with plasma D3 levels higher than 5 ng/ml. In addition, a hyperbolic regression curve was obtained between the plasma D3 and 25-OH-D3 ratio. These results may indicate that a possible negative feedback homeostatic mechanism exists between plasma levels of D3 and 25-OH-D3, but only with low plasma levels of D3. PMID- 6286380 TI - [Antibody to 1-24 ACTH and hypofunction of the adrenal cortex in a patient who was unilaterally adrenalectomized for Cushing's syndrome (author's transl)]. AB - We report here on a patient who was unilaterally adrenalectomized for Cushing's syndrome, and who developed antibodies to 1-24 ACTH. A 49-year-old nurse had undergone right adrenalectomy for adrenal adenoma. After surgery, she was treated with 0.5 mg of 1-24 ACTH-Z together with glucocorticoid replacement therapy for 40 days. Thereafter she was given 0.25-0.5 mg of ACTH-Z every other day for 4 months. ACTH-Z was then stopped for a year but glucocorticoid therapy was continued. About one year prior to this admission, 1 mg of ACTH-Z was again initiated 1 to 2 times a week. Glucocorticoid therapy was not withdrawn during the four years after adrenalectomy. She was admitted for the purpose of withdrawal of glucocorticoids. Her serum was found to bind labeled ACTH. This labeled ACTH was competitively displaced from binding by unlabeled hormones. Finally, reaction with specific antihuman Ig demonstrated an antibody of the IgG class. The titer of the antibodies gradually decreased after the discontinuation of ACTH-Z, but it is still present in measurable quantity in her serum. The clinical significance of the circulating anti-ACTH antibody in her serum is discussed. PMID- 6286381 TI - Characterization and localization of the transferrin receptor on rat reticulocytes. AB - 1. A further characterization and localization of the membrane receptor for transferrin on rat reticulocytes is described. PAGE studies with a purified membrane complex B2, from which the functional role in transferrin binding and iron uptake has been shown previously, showed that the transferrin receptor is localized on a membrane protein with a mol. wt of approximately 70-80.10(3). 2. Selective solubilization of the rat reticulocyte membrane has shown that this receptor protein belongs to one of the minor integral membrane polypeptides, embedded in the lipid bilayer of the membrane. 3. Proteolipid complexes, glycolipids and sialoglycoproteins of the rat reticulocyte membrane play no direct role in the binding capacity of the receptor. PMID- 6286382 TI - Identification and solubilization of a cAMP-independent protein kinase from mouse L-cell smooth membranes. AB - 1. Smooth membranes have been prepared from mouse L-cells and found to contain an endogenous protein kinase activity. 2. The enzyme(s) responsible for this activity use ATP, but no other nucleoside triphosphates, to phosphorylate endogenous membrane proteins as well as exogenously-added protein substrates such as phosvitin and casein. 3. Mg2+ is required for enzyme activity, maximal activity is observed at pH 7.5-8.0 and the kinase is not dependent on, or stimulated by, cyclic 3'-5' AMP. 4. The kinase activity is not decreased by the Walsh heat-stable inhibitor of cyclic 3'-5' AMP-dependent protein kinases. 5. Fifty percent or more of the membrane-associated kinase activity can be solubilized by extracting membranes with buffer containing 0.6 M NaCl. 6. The solubilized enzyme resembles the membrane-associated activity in its Mg2+ requirement, pH optimum and independence of cyclic 3'-5' AMP. 7. Phosvitin and casein are better exogenous substrates than histones or protamine for phosphorylation by the enzyme in either the membrane-associated or solubilized state. PMID- 6286383 TI - Influence of DL-ethionine on the NAD content and on the activity of the poly(ADPR)synthetase in the rat liver. AB - 1. Ethionine, an analogue of methionine, is known to cause a number of serious changes. In this study, its influence on the NAD + NADH2 content and on the activity of the poly(ADPR)synthetase in rat liver has been investigated. 2. DL Ethionine decreases the content of NAD + NADH2 in adrenalectomized, but not in normal animals. 3. After injection of nicotinamide the NAD + NADH2 content in the liver of adrenalectomized rats is much higher than in that of normal ones. 4. DL Ethionine diminishes the NAD synthesis caused by nicotinamide in adrenalectomized animals and increases it in normal ones. DL-Tryptophan induces only a slight NAD synthesis, which is reduced in adrenalectomized animals by thionine. 5. 1 Methylnicotinamide slightly decreases the NAD concentration in normal animals, but increases it considerably in adrenalectomized ones. DL-ethionine reduces the NAD + NADH content in adrenalectomized animals in the presence of 1 methylnicotinamide. 6. The activity of poly(ADPR)synthetase of in-vitro tested nuclei is slightly increased after treatment with DL-ethionine. PMID- 6286384 TI - Effects of fluoride on sugar phosphate phosphohydrolase from Escherichia coli and Neisseria meningitidis. AB - The purified sugar phosphate phosphohydrolase from E. coli and N. meningitidis were used for our study and biphasic responses were observed in the presence of fluoride. 2. From the kinetic studies of fluoride inhibition, the enzymes from both bacteria appear to catalyze the hydrolysis of alpha-D-aldohexose 1-phosphate and alpha-D-aldohexose 6-phosphate at two distinct catalytic sites. PMID- 6286385 TI - Dinucleosidetetraphosphatase from Ehrlich ascites tumour cells: inhibition by adenosine, guanosine and uridine 5'-tetraphosphates. AB - 1. An enzyme has been partially purified from Ehrlich ascites tumour cells which specifically hydrolyses dinucleosidetetraphosphates, with Km values of around 2 microM. The products of the hydrolysis are the corresponding nucleoside tri- and monophosphates. Dinucleoside Tri- and diphosphates were not substrates of the reaction. 2. The enzyme requires Mg2+ or Mn2+, is maximally active at a pH value of approx. 7.5 and has a mol, wt of 19,800 as estimated by filtration on Sephadex G-75. Nucleoside mono-, di- and triphosphates were competitive inhibitors of the reaction with Ki values in the 0.1 mM range. 3. Particularly relevant is the inhibition of this enzyme by adenosine and guanosine 5'tetraphosphates. In the course of this investigation, the presence of uridine 5'-tetraphosphate was detected in a commercial preparation of UTP. Adenosine, guanosine and uridine 5' tetraphosphates were very strong inhibitors of the reaction with Ki values in the nM range. PMID- 6286386 TI - Nucleoside triphosphate syntheses by chromatin bound enzymes from Morris hepatoma 9121 nuclei. AB - 1. Enzymatic syntheses of deoxyribo- and ribonucleoside triphosphates were observed with purified, chromatin bound enzyme C from Morris Hepatoma 9121 nuclei. 2. Intermediate substrates were nucleoside mono- and diphosphates for enzyme C. 3. The three phosphate residues of ATP serving as energy-rich phosphate donor were successively transferred to a nucleoside substrate by enzyme C. PMID- 6286387 TI - Studies on the mechanisms of LH receptor control by FSH. PMID- 6286389 TI - Modulation of prolactin receptors in cultured rat granulosa cells by FSH, LH and GnRH. AB - The hormonal modulation of prolactin (PRL)-binding capacity of rat granulosa cells was studied. Granulosa cells obtained from immature, hypophysectomized, estrogen-treated rats were cultured for 2 days in a serum-free medium in the presence of various hormones. FSH treatment in vitro stimulated granulosa cell PRL-binding capacity by approximately 4-6-fold in a dose-dependent manner. Concomitant treatment with 10-8 M GnRH inhibited the FSH-induced increase in PRL binding capacity by 64%. In contrast, the inhibitory effect of GnRH was blocked by concomitant treatment with 10-6 M of a GNRH antagonists, [D-pGlu1, D-Phe2, D Trp3,6]GnRH. PRL-binding capacity was also increased (approximately 2-fold) by in vitro treatment with cholera toxin (10 microgram/ml). In granulosa cells pre treated with FSH in vitro for 2 days, hCG treatment for 2 additional days stimulated PRL-binding capacity in a dose-dependent manner (approximately 2 fold). Likewise, treatment with LH (100 ng/ml) also stimulated PRL-binding capacity by approximately 2-fold. These in vitro studies demonstrated that gonadotropins (FSH, LH and hCG) directly enhanced PRL binding by granulosa cells, whereas GnRH inhibited FSH action. PMID- 6286388 TI - Stimulation and inhibition by LHRH analogues of cultured rat Leydig cell function and lack of effect on mouse Leydig cells. AB - The effect of 2 luteinizing hormone-releasing hormone (LHRH) analogues(10-8-10-6 M) on the functional activity (testosterone and cyclic AMP production and [125I]hCG binding) of purified mouse Leydig cells in culture was examined. The analogues were found to have no significant effect on the cells over a period of 3 days. No specific binding of a labelled analogue to impure or pure mouse Leydig cells could be detected. In contrast high levels of specific binding to impure rat interstitial cells occurred. Centrifugation of the rat interstitial cells on 0-90% Percoll gradients showed that the LHRH analogue bound specifically to the active lutropin-responsive Leydig cells. The purified rat Leydig cells were cultured in the presence of LHRH analogue (ICI 118630) (10-7 M) and after an initial lag period (2h) a marked stimulation of testosterone production occurred over a 32-h period (up to 400 ng/10(6) cells). The response to LH alone increased with time in culture up to 10 h, and the LHRH analogue enhanced this LH stimulated testosterone production. When the cells were cultured for longer time periods (24 h) the LHRH analogue was found to inhibit LH-stimulated testosterone production at all concentrations of LH used (p less than 0.01). The LHRH analogue had no consistent effect on LH-stimulated cyclic AMP production, although when added alone, cyclic AMP production was increased. These results show that LHRH analogues do not bind to or have any detectable effect on mouse Leydig cells in vitro. However, LHRH analogue does bind specifically to purified rat Leydig cells. After a short lag period the analogue stimulates testosterone production which turns to inhibition after 20 h in culture. PMID- 6286390 TI - Studies on the dynamics of adrenocortical responses to ACTH in the rat. PMID- 6286391 TI - Developmental regulation of multiple forms of UDPglucose pyrophosphorylase of Dictyostelium. PMID- 6286392 TI - Effects of betazole hydrochloride and cyclic AMP on the pepsinogen secretion by rabbit gastric mucosa in organ culture. AB - The effects of betazole hydrochloride, dibutyryl cyclic AMP (DB-cyclic AMP) and betazole hydrochloride plus aminophylline on pepsinogen secretion by rabbit gastric mucosa were studied in organ culture. Betazole hydrochloride alone did not stimulate pepsinogen secretion at the concentrations of 10(-8), 10(-6), 10( 5) and 10(-4) M. However, 10(-3) M DB-cyclic AMP produced a significant stimulation of pepsinogen secretion into the culture medium when compared with the control. In the presence of 3 x 10(-3) M aminophylline, betazole hydrochloride in the concentration of 10(-5) and 10(-4) M stimulated pepsinogen secretion into the culture medium, and the magnitude of this increase was 2.0- and 2.8-fold, respectively, compared with the control. Aminophylline alone could not change pepsinogen secretion into the culture medium. These results suggested that the pepsinogen secretion, stimulated by betazole hydrochloride, was mediated by cyclic AMP in the chief cells. PMID- 6286393 TI - Characterization of secretin and vasoactive intestinal peptide receptors in rat pancreatic plasma membranes using the native peptides, secretin-(7-27) and five secretin analogues. AB - A comparison has been made of the ability of vasoactive intestinal peptide (VIP), secretin, secretin analogues, and secretin-(7-27) to stimulate adenylate cyclase in rat pancreatic plasma membranes. A parallel study of the capacity of peptides of the VIP-secretin family to compete with 125I-VIP for binding to the same plasma membranes was conducted. This allowed a classification of VIP-secretin receptors into three subtypes: (1) VIP-preferring receptors; (2) high-affinity secretin receptors, and (3) low-affinity secretin receptors. The properties of secretin at high-affinity secretin receptors were likely to reflect a contribution of membranes from centroacinar and duct cells. PMID- 6286394 TI - Reduced Coxsackie antibody titres in type 1 (insulin-dependent) diabetic patients presenting during an outbreak of Coxsackie B3 and B4 infection. AB - The potential role of antecedent viral infection in the pathogenesis of Type 1 (insulin-dependent) diabetes was investigated by measuring antibody titres to several viruses in serum obtained at the time of diagnosis of diabetes. An outbreak of Coxsackie B4 infection followed by a wave of Coxsackie B3 and B5 infections occurred in Seattle during the time viral serology was obtained in the diabetic patients. Antibody titres to Cocksackie B5 and Influenza A and B viruses were comparable in diabetics and matched control subjects, but antibody titres to Cocksackie B3 and B4 were lower in the diabetics and a low antibody titre to Coxsackie B3/B4 was associated with a significantly increased relative risk of diabetes. PMID- 6286395 TI - Relationship between rate of digestion of foods and post-prandial glycaemia. AB - The amount of carbohydrate released at 1 and 5 h by digestion in vitro of 2 g carbohydrate portions of 14 foods by human digestive juices was compared with the area under the 2-h blood glucose response curve when 50 g carbohydrate portions were fed to groups of five to ten healthy volunteers. A significant relationship was found between the amounts of sugars and oligosaccharides liberated at 1 and 5 h and the food blood glucose area expressed as a percentage of the blood glucose area for 50 g glucose (r = 0.8627 and 0.8618, p less than 0.001). A significant relationship was also found between the glycaemic index and the food fibre content (p less than 0.02) and between the glycaemic index and the glucose trapping capacity of the foods (p less than 0.05). Legumes as a group liberated 56% less sugars and oligosaccharides (p less than 0.01) than the eight cereal foods over 5 h. It is suggested that such studies in vitro may help to identify food of use for diabetic patients, and at the same time throw further light on factors which affect post-prandial glycaemia. PMID- 6286397 TI - Differentiated mouse epithelial cell line with hepatocyte characteristics. AB - An epithelial cell line, 3105, with an unusual growth pattern has been derived from the liver of an (NZB x NZW)F1 mouse. When confluent, it forms a monolayer of closely packed cells interspersed with holes that do not fill in during cultivation. By electron microscopy, the line has tight and intermediate junctions as well as desmosomes typical of epithelial cells. It produces several enzymes normally present in liver including hydroxymethylglutaryl-coenzyme A (HMG CoA) reductase, glucose-6-phosphatase, and alkaline phosphatase; has cytochromes P-450 and b5; and spontaneously release xenotropic but not ecotropic endogenous mouse type C viruses. Inoculation of the cell line into athymic nude mice gives rise to benign cysts in 2-3 months. This mouse epithelial line with hepatocyte characteristics should be helpful to investigators as a cell model of normal liver cell differentiation. PMID- 6286396 TI - Increased dose-response relationship of liver plasma membrane adenylate cyclase to glucagon stimulation in diabetic rats. A possible role of the guanyl nucleotide-binding regulatory protein. AB - In view of controversial findings regarding the mechanism for the increased intracellular hepatic cyclic 3':5' adenosine monophosphate levels in diabetic rats, we studied the dose-response relationship of the adenylate cyclase to glucagon stimulation in severely diabetic and in diabetic, insulin-treated rats. An enhanced response to glucagon and an additional augmenting effect of guanosine triphosphate on hormonal stimulation of the adenylate cyclase activity were found in diabetes which were reversible with insulin treatment. The results suggest a role of the regulatory guanyl nucleotide-binding protein in diabetes leading to an increased dose response relationship of the hepatic adenylate cyclase system to glucagon. PMID- 6286398 TI - Herpesviruses in metastatic Lucke renal adenocarcinoma. AB - The etiologic agent of the renal adenocarcinoma of leopard frogs, Rana pipiens, is the Lucke tumor herpesvirus (LTHV). The virus is easily detected with thin section electron microscopy in primary tumors of frogs which have been exposed to a cold environment. Several spontaneous metastatic nodules and a large primary tumor were detected at autopsy of a frog which had been maintained at 4 degrees C for 73 days. LTHV was found not only in the primary tumor, as previously reported, but also was present in metastatic tumor cells in the liver, fat body, and bladder. The presence of LTHV in metastatic cells demonstrates that the differentiated state of primary Lucke tumor cells is retained in its metastatic colonies even at the fine structure level revealed by electron microscopy. PMID- 6286399 TI - Polyoma virus mutants as probes of variety among mouse embryonal carcinoma cell lines. AB - Embryonal carcinoma (EC) cells are resistant to infection with polyoma virus and become permissive when allowed to differentiate. Polyoma host-range (PyEC) mutants have been selected on two embryonal carcinoma cell lines, PCC4 and F9, which differ in their differentiation potential, both in vitro and in vivo. Since PyEC mutants selected on one line failed to develop, or developed only poorly on the other, we used these two classes of mutants as probes towards several EC lines which differed in their origin and differentiation properties. From their susceptibility to either mutant, and from the effect of temperature upon the efficiency of infection, we inferred a classification of these teratocarcinoma cell lines, which is an agreement with a previous one based upon metabolic coupling. We also discussed results indicating that different EC cell lines might present steps in the process of cell determination at the embryonic level. PMID- 6286401 TI - Measurement of gallbladder emptying sequentially using a single dose of 99mTc labeled hepatobiliary agent. AB - The gallbladder emptying response to a single large (20 ng/kg) and four equally divided sequential small (5 ng/kg each) doses of octapeptide of cholecystokinin at 20-min intervals was measured in a paired study in six normal subjects noninvasively and quantitatively by a nongeometric method using a gamma camera, computer, and 99mTc-labeled hepatobiliary agent. The gallbladder mean (+/- SD) ejection fraction after a single large dose (20 ng/kg) of octapeptide of cholecystokinin was 34 +/- 24% and after four sequential small (5 ng/kg each) doses was 42 +/- 14%, 45 +/- 28%, 37 +/- 22%, and 32 +/- 27%, respectively (p greater than 0.05). The results of this study indicate that the first part of the 5 ng/kg sequential octapeptide of cholecystokinin dose is as effective as the single 20-ng/kg dose. The gallbladder emptying requires continued presence of high levels of octapeptide of cholecystokinin in the serum closer to levels that initiate contraction and in the absence of which the gallbladder ceases to empty further despite the fact it has inherent capacity to do so. Each of four equal octapeptide of cholecystokinin doses given sequentially elicits, on the average, an equal degree of emptying response. The method has potential for application in the study of the pharmacological effects of drugs on the biliary dynamics using a single dose of 99mTc-labeled hepatobiliary radiopharmaceutical. PMID- 6286400 TI - Impaired metabolism of methionine in severe liver diseases. I. Clinical and pathophysiological significance of elevated serum methionine levels. AB - Serum methionine levels increased to a greater extent in patients with severe liver diseases such as fulminant hepatitis and liver cirrhosis with and without hepatic encephalopathy. However, the concentrations remained unchanged in non encephalopathic cirrhotic cases associated with hepatocellular carcinoma, and their serum methionine levels increased only moderately even at the time of encephalopathy. At least two different mechanisms of serum methionine elevations, possibly due to release from injured hepatocytes or diminished catabolisms of this amino acid in the damaged liver, could be differentiated; the former would be involved mainly in fulminant hepatitis and the latter in liver cirrhosis. A methionine-loading test performed in cirrhotic patients supported the validity of these considerations. No significant increase of serum methionine levels in cirrhotic patients with hepatocellular carcinoma was observed, possibly by remarkable consumption of this amino acid in hepatoma tissues. During the clinical course of several patients, serial determinations of serum methionine concentrations indicated that the levels varied depending upon alterations in the pathophysiological state of the damaged liver; much higher levels were observed concomitantly with decompensated signs such as ascites, jaundice and hepatic encephalopathy. These results suggest that monitoring of serum methionine levels would be very valuable, especially for judging prognosis and predicting hepatic encephalopathy in severe liver disease. PMID- 6286402 TI - Sustained pectin ingestion delays gastric emptying. AB - The effects of sustained fiber ingestion on gastric emptying glucose tolerance, hormone responses, and jejunal absorption of glucose and lysine were studied in healthy volunteers. Subjects were placed on a low-fiber (3 g) diet for 2 wk, followed by 4 wk of an isocaloric diet supplemented with 20 g/day of either apple pectin (7 subjects) or alpha-cellulose (6 subjects). At the conclusion of each dietary period subjects ingested a low-fiber breakfast surface-labeled with 99mtechnetium sulfur-colloid. Gastric emptying half-time, plasma glucose, calcium, phosphorus, insulin, glucagon, gastrin, human pancreatic polypeptide, and motilin were determined. Gastric emptying half-time was prolonged approximately twofold after pectin supplementation (p less than 0.005) and returned to normal 3 wk after discontinuing pectin supplementation. Cellulose supplementation did not alter the gastric emptying rate. Plasma glucose, calcium, phosphorus, and hormonal responses to the meal were unchanged after either pectin or cellulose supplementation. Pectin ingestion did not impair intestinal absorption of glucose or lysine. In contrast to sustained cellulose ingestion, sustained pectin ingestion slows the gastric emptying rate; the mechanism underlying this adaptive effect is unknown. PMID- 6286404 TI - [Results of the treatment of trophoblast tumours (author's transl)]. AB - Within 10 years 24 patients were treated for choriocarcinoma. Experience with new methods of diagnosis, treatment and followup was gained. 11 patients were considered to be in a low risk group and 13 patients were categorized as high risk patients. The treatment was primarily conservative. Patients with low risk received single treatment with methotrexate. In some cases in the high-risk group combination chemotherapy or polychemotherapy was used. In addition to the clinical findings, the results of the treatment are monitored with serial beta HCG determinations. Of the 11 patients in the low-risk group all well and free of recurrence. Of the high-risk group 6 patients died. Of the total of 24 patients with choriocarcinoma 18 (75%) are well and free of recurrence. PMID- 6286405 TI - Effects of alpha and beta adrenoceptors on branchial cAMP level in seawater mullet, Mugil capito. PMID- 6286406 TI - Testosterone inhibition of FSH action on testicular 32P uptake in white leghorn cockerels. PMID- 6286407 TI - Vitamin D3-induced hypercalcaemia in male catfish, Clarias batrachus. PMID- 6286403 TI - [Determination of cytomegalo-, herpes simplex- and rubellavirus-antibodies resp. during perinatal period by Elisa-technique (author's transl)]. AB - Cytomegalovirus (CMV)-infections attract increased attention among viral infections in the pre- und perinatal period resp. The decreasing cell mediated immunity by the end of pregnancy may favour conditions for primary infections or reactivations with CMV. With the Elisa-technique we looked for CMV-specific antibody status in the perinatal period with special respect to striking titers. The same serum samples were screened for antibody status to Herpes simplex virus (HSV) type 1 specific IgG and rubellavirus specific IgG resp. 67% were positive for CMV-IgG, 90% for HSV-type 1-IgG and 93.5% for rubellavirus IgG. CMV-specific IgM antibodies were found in 3.6% but only 1.5% were suspicious for recent CMV infections according to higher CMV specific IgM titers. PMID- 6286408 TI - ACTH activity in the pituitary and brain of the least brook lamprey, Lampetra aepyptera. PMID- 6286409 TI - Isolation and characterization of Streptomyces fradiae plasmids which are prophage of the actinophage phiSF1. AB - Two plasmids, designated pUC1 and pUC13, have identified as different prophage states of the Streptomyces fradiae actinophage phiSF1. Plasmids pUC1 and pUC13, 51.1 Md, are identical as determined by restriction endonuclease analysis and other criteria. They differ in that while pUC1 specifies "type A" plaques, characterized by a narrow zone of growth inhibition around a transformant colony, pUC13 specifies "type B" plaques, characterized by concentric rings of growth and growth inhibition around a transformant colony. Additionally cultures carrying pUC13 release antinophage phiSF1 at a frequency about 10 3-10 4 times that pUC1 containing cultures. Restriction enzyme digestion, transfection, and transformation analyses show that phiSF1 DNA, 55.7 Md, is about 8-10% larger than pUC13 and appears to be circularly permuted and terminally redundant. The pUC1 and pUC13 prophage are interconvertible. PMID- 6286410 TI - The cloning of the Escherichia coli K-12 deoxyribonucleoside operon. AB - A 6.1-kb EcoRI DNA fragment containing the four structural genes (deoC, deoA, deoB, deoD) of the deoxyribonucleoside operon has been cloned into the plasmid pMFS53. By use of a unique, asymmetrically positioned HindIII site on the 6.1 kb insert, plasmids containing the deoC,deoA genes (pMFS50) or the deoB,deoD genes (pMFS55) have been constructed. Enzyme assays performed on extracts prepared from clones harboring pMFS53, pMFS50 or pMFS55 revealed that each clone possessed amplified deo enzyme levels and that the spectrum of enzyme amplification corresponded to the genetic composition of the plasmids carried by each clone. A plasmid (pMFS50l) having functional deoA, deoB and deoD genes but devoid of the deo regulatory region and a portion of the deoC structural gene has been isolated following treatment of BamHI cleaved pMFS53 and BAL31 nuclease. Comparison of the deo enzyme levels for clones harboring pMFS53 and pMFS501 suggest that plasmid pMFS53 possesses a functional deo regulatory region in addition to the four structural genes of the operon. PMID- 6286412 TI - Molecular cloning of AKR xenotropic murine leukemia virus unintegrated proviral DNA. AB - Suprahelical proviral DNA of AKR xenotropic murine leukemia virus was purified from agarose gels and cloned in lambda Charon 28 DNA (BamHI sites). Nine viral DNA recombinants were identified and mapped with 12 restriction endonucleases. Three calsses of cloned viral DNA inserts were found: (1) Six inserts were apparently full-length 9.0-kb DNA with tandem long terminal repeat (LTR) elements; (2) two inserts contained DNAs with deletions in or adjacent to the LTR regions; (3) a single isolate contained an inversion of 2.3 kb around the LTR in the envelope gene. PMID- 6286411 TI - Analysis of plasmid genome evolution based on nucleotide-sequence comparison of two related plasmids of Escherichia coli. AB - Plasmid Rsc13, a small derivative of the plasmid R1, contains a region necessary for replication as well as a complete copy (4957 bp) of the ampicillin resistance transposon, Tn3. We determined the nucleotide sequence of the replication region of Rsc13 to be 2937 bp and then compared this region (designated the 2.9-kb region) to the analogous region of pSM1, a small derivative of the plasmid R100 which has common ancestry with R1. Rsc13 and pSM1 were 96% homologous in this 2.9 kb region except for a discrete region of about 250 bp which showed only 44% homology. The sequence and distribution of nucleotide substitutions between Rsc13 and pSM1 supported a map of possible genes and sites which have previously been seen in the replication region of Rsc13 and pSM1 which showed only 44% homology. Analysis of the amino acid sequence and predicted conformation of the two RepA2 polypeptides, however, suggested that they were very similar. We proposed that the repA2 region of R1 and R100 was replaced by a substitution of a short DNA segment from another plasmid which was evolutionarily related to R1 and R100 but had more divergence. This event may have been mediated by a mechanism similar to that of gene conversion as described in eukaryotic systems. PMID- 6286413 TI - Restriction map of bacteriophage SP02: ordering the SacI fragments and identification of the DNA termini. PMID- 6286414 TI - Plasmid cloning vectors resistant to ampicillin and tetracycline which can replicate in both E. coli and Haemophilus cells. AB - We have constructed two plasmid vectors, pHCV5 and pHVTl, which will replicate both in Haemophilus and in Escherichia coli. Both contain the ampicillin resistance gene and the replication origin from a Haemophilus plasmid, pRSF0885. Both also contain the pBR322 origin and therefore can be amplified in E. coli by chloramphenicol treatment. The plasmid pHCV5 contains the tetracycline-resistance gene of pBR322, and pHVT1 contains the analogous region from the transposon Tn10. PMID- 6286415 TI - Partial characterization of a small, multiple-copy plasmid from Streptomyces espinosus and the derivation of a high copy-number deletion mutant. AB - An organism classified as Streptomyces espinosus was found to carry an approx. 9.2-kb plasmid. This plasmid, designated pUC6, has a copy number of 30-40 per host genome equivalent. Plasmid pUC1061, a copy-number mutant of pUC6, was isolated after in vitro deletion of an approx. 2.0-kb XhoI restriction fragment. Plasmid pUC1061 has a copy number of 500-600. Plasmid pUC1061 appears to be incompatible with pUC6 and will transform a pUC6-containing culture at a frequency of approx. 1%. The sizes, restriction maps and copy numbers of plasmids pUC6 and pUC1061 indicate these may be valuable vectors for gene cloning Streptomyces. PMID- 6286416 TI - The site-specific deletion in plasmid pBR322. AB - The formation of a deletion derivative of plasmid pBR322, designated pBR322 delta 1, was observed during cloning of various eukaryotic DNAs, when the BamHI site of the plasmid vector was used for construction of the recombinant molecules. The restriction analysis of six independently isolated pBR322 delta 1 plasmids allowed establishment of their complete identity. Similar deletion derivatives were also formed as a result of transformation of Escherichia coli cells by the linear form of vector pBR322 produced by BamHI cleavage, but not by SalI or HindIII. The endpoints of the deletion in one of the pBR322 delta 1 plasmids occurred at positions 375 and 16666 bp from the EcoRI site, as determined by sequence analysis. Formation of pBR322 delta 1 is most probably due to site specific recombination between the sequence in the 1666-1670 bp region and the BamHI end of the linear pBR322 molecule. THe deletion was not controlled by the recA system of the host bacteria. PMID- 6286417 TI - Cloning of the complete human cytomegalovirus genome in cosmids. AB - Purified virion DNA (155 X 10(6) Mr) of human cytomegalovirus (CMV) strain Ad169 was partially cleaved with restriction endonucleases HindIII and EcoRI and cloned in the respective cleavage sites of cosmid pHC79. A complete gene library was established in a set of clones containing the viral DNA in long overlapping segments. Restriction maps for HindIII (29 fragments) and EcoRI (36 fragments) were constructed from the linkage of cosmid-cloned fragments, from double digestions of cloned DNA, and from blot hybridization of labeled cloned viral DNA with restriction fragments of virion DNA and singly or doubly cleaved cosmid clones. PMID- 6286418 TI - Bvui: a site-specific endonuclease from Bacillus vulgatis. AB - A site-specific endodeoxyribonuclease was partially purified from an extract of osmotically shocked spheroplasts of a Bacillus vulgatis strain; the enzyme has been designated BvuI and its activity was characterized. The locations of BvuI generated cleavages on bacteriophage lambda and M13 derivatives mp7, mp8 and mp9, SV40 and PBR322 DNA molecules were determined. BvuI was shown to recognize the DNA sequence decreases 5'-G-Pu-G-C-Py-C-3' 3'-C-Py-C-G-Pu-G-5' increases and cleaves it at the positions indicated by arrows. Two identical BvuI recognition sites separated by fourteen nucleotide pairs were shown to occur within the tetracycline resistance gene of PBR322; BvuI should be useful for molecular cloning in that plasmid vector. PMID- 6286419 TI - Mapping of the plasmid (pLQ3) from Achromobacter and cloning of its cephalosporinase gene in Escherichia coli. AB - We have constructed a physical map of the plasmid pLQ3 which was originally isolated from Achromobacter and which codes for a beta-lactamase. The enzyme specified by pLQ3 is expressed in Escherichia coli and is unusual in that it is a cephalosporinase, an enzyme usually coded for by chromosome. Plasmid pLQ3 is 12.4 kb in length and has a unique Bam HI site and two BglII sites. From a BamHI + BglII double digest of pLQ3, we have constructed a "shortened" plasmid, pLQ10, in which a 2.96-kb fragment is deleted. We have constructed a clone, pLQ22, in which a 3.27-kb fragment of pLQ3, carrying the beta-lactamase gene, is inserted into the BamHI site of pACYC184. By "comparative mapping" of single and multiple digests of each of these plasmids, we have been able to locate the cleavage sites for PstI, which makes seven cuts in pLQ3. PMID- 6286420 TI - Effect of lexA and ssb genes, present on a uvrA recombinant plasmid, on the UV survival of Escherichia coli K-12. AB - The recombinant plasmid pJA01 contains, besides the uvrA gene, the genes lexA, ubiA and ssb. This plasmid does not fully complement a uvrA mutation in a Rec+ background. Plasmids which contain the uvrA and ssg genes, but not the lexA gene, show a higher but still only partial complementation. Full complementation achieved when the ssb gene us inactivated by insertion of Tn5. Furthermore, it appears that the presence of the ssb gene on a multicopy plasmid sensitizes wild type cells to UV light. The effect of Ssb (single-strand DNA binding protein) overproduction on UV survival is discussed. PMID- 6286421 TI - Isolation of two restriction endonucleases from Chloroflexus aurantiacus (CauI, CauII). PMID- 6286422 TI - The sequence specificity of endonucleases CauI and CauII isolated from chloroflexus aurantiacus. AB - The type II restriction enzymes CauI and CauII, isolated from Chloroflexus aurantiacus, recognize and cleave (at the position indicated by an arrow) the sequences G decreases G A/T CC and CC decreases G/C GG, respectively. These conclusions are supported by the results from restriction site mapping, sequence analysis by partial chemical degradation, end-group analysis after lambda exonuclease treatment and computer-assisted comparison of DNA sequence data. PMID- 6286423 TI - Cloning SV40 HindIII restriction fragments into M13 bacteriophage. PMID- 6286424 TI - Identifying and treating psychiatric disorders in cancer patients. PMID- 6286425 TI - [Tasks and methodological approaches to the hygienic study of carcinogens in the atmosphere]. PMID- 6286426 TI - [Possible benzo(a)pyrene pollution of the soil and air in an area of agricultural machinery operation]. PMID- 6286427 TI - [Staging and grading experiences in ovarian carcinoma and their prognostic value]. PMID- 6286428 TI - [Detailed description of the primary flexor tendon suture in the digital canal]. PMID- 6286429 TI - [Glomus tumor, mistaken for an inflammation for 4 years]. AB - The difficulties encountered in diagnosing a glomus tumor are demonstrated by means of the case of a 34 year old female patient. The available literature specifies that the course of the disease may extend over a time of up to 40 years. Troubles over a period of time of only a few months are rather exceptional. In the present case, taking a sample excision from a distinctly clubbed finger helped to clarify the diagnosis. In view of the long course of the symptoms as well as the gross clinical and suspect radiological findins, and eventually malignant disease could not be excluded from the differential diagnosis. PMID- 6286430 TI - [Scinti-angiographic investigation of aortic dissection (author's transl)]. AB - We perform scinti-angiography regularly in suspected cases of dissecting aneurysms. In our experience, the dissection is well demonstrated by this method. In correctly selected cases, scinti-angiography is a method which is well tolerated by the patient and which carries very little risk, since the complications due to contrast media are absent. PMID- 6286431 TI - [Percutaneous transhepatic portal vein catheterisation in the diagnosis of hormone-producing tumours in the splanchnic area (author's transl)]. AB - During 1980, percutaneous transhepatic portal vein catheterisation was carried out in eight patients with suspected hormone-producing tumours in an attempt to localise these. Seven patients with hyperinsulinism were suspected of having an insulinoma and one a gastrinoma. There were no complications following the catheterisation, and difficulties were reduced by the use of a special catheter. The blood samples were examined by radio-immunological methods for insulin or gastrin. On the basis of localised rise in hormone level, it was possible to diagnose three insulinomas and one gastrinoma in the pancreas. The beta-cell tumours were removed by enucleation or segmental resection, the gastrinoma by a Whipple's operation. Three patients showed an insulin gradient and sub-total pancreatectomies were performed. In one patient there was no definite abnormality in the insulin level and an operation has so far not been performed. Our experience indicates that this technique is difficult, bu is nevertheless a reliable method for localising hormone-producing tumours and surgery should not be carried out without it. PMID- 6286432 TI - [Lipiodol embolism of the liver on a computer tomogram]. PMID- 6286433 TI - Carcinoma of the gall bladder: a CT diagnosis. PMID- 6286434 TI - [Ultrasonic mammography: its importance in integrated diagnosis of breast diseases (author's transl)]. AB - Sonography of the breast is gaining in importance with the spread of diagnostic methods based on ultrasound in general and the increasing technical perfection of the equipment in particular. Over and above this, the increase in incidence of carcinoma of the breast at an increasingly early age represents a permanent challenge to achieve further optimisation of diagnostic methods. The following conclusions have been arrived at after evaluating 493 ultrasonic examinations of the breast, 212 of which were verified histologically (90 carcinomas): 1. If the indication is precise, sonography possesses a high degree of sensitivity in diagnosing any changes occurring in the breast. 2. The method offers particular advantages in patients with manifest signs having a dense parenchyma; in fact, with such patients sonography is often superior to mammography. 3. The additional use of sonography in diagnosis of diseases of the breast improves the results and often facilitates a less aggressive procedure (insofar as biopsies can be omitted). The main drawbacks are the great amount of time required for performing the examination, and the problem of documentation. This has been the major obstacle to using sonography in screening. The article discusses the indications, the typical sonographic criteria and problems of differential diagnosis in relation to the most frequently encountered diseases of the breast. PMID- 6286435 TI - [Measurements of the intracranial CSF spaces by computer tomography (author's transl)]. AB - A method for quantifying the CSF compartments in cranial computer tomograms is described. The method is based on an analysis of the histogram curve, the proportion of area and thickness of the liquor spaces being related to the total area below the curve. In order to isolate the skull contents, a CT masking technique is used. The practicability and usefulness of the method in clinical practice was tested in 59 normal patients and in eleven patients with rapidly progressing brain atrophy. PMID- 6286436 TI - [Daily menus for diet therapy of liver cirrhosis]. PMID- 6286438 TI - [The renaissance of theophylline therapy]. AB - Deeper insight into the pharmacodynamics and pharmacokinetics of theophylline during the past 10 years led to the wide-spread use of the substance in the treatment of obstructive airway diseases. The effect of theophylline can be explained by different mechanisms of action, namely increase in cAMP and prostaglandins, redistribution of calcium, and blockade of purinergic receptors. Besides the known antiobstructive activity of theophylline ethylenediamine, its influence on the respiratory muscles is an essential clinical quality. The results of pharmacokinetic studies and routine determination of theophylline levels contributed to greatly improve the efficacy of theophylline therapy. At adequate doses, theophylline or theophylline ethylenediamine in sustained-release form constitutes the most effective long-term treatment of chronic asthma today. The pharmacological properties of ethylenediamine reinforce some of the more important effects of theophylline and improve tolerance. Combining the substance with beta-sympathomimetics elicits additive effects, while side effects are kept low. PMID- 6286437 TI - [Tumor-associated antigens in bronchial cancer]. PMID- 6286439 TI - [Amebiasis]. AB - Infections with Entamoeba histolytica do not necessarily cause disease in those infected. The parasite may act as commensal (cysts living in the bowel) or it may cause a broad spectrum of clinical illness. Some of the factors causing overt disease are poorly understood. An acute amebic dysentery is accompanied by bloody stools, abdominal pain and indigestion. The most important extraintestinal complication of an amebic infection is a liver abscess causing severe pain, fever, nausea and vomiting. The diagnosis of an amebic infection is based upon isolation of the parasite from the stools. Extraintestinal amebiasis is diagnosed - apart from the clinical picture - by serology. For treatment of intestinal amebiasis so-called contact-amebicides can be recommended. An amebic abscess of the liver usually responds well to dehydroemetine, metronidazole or any other derivative or imidazole and chloroquine. Surgical treatment of amebic liver abscess is only required if complications arise. PMID- 6286440 TI - [Immunological response of patients with herpes zoster (author's transl)]. PMID- 6286441 TI - Plasma androgens in children and adolescents. Part II. A longitudinal study in patients with hypopituitarism. AB - In this prospective longitudinal study, plasma androgen levels were determined during 1-7 years in 45 patients aged 5.6-23.8 years with either isolated growth hormone (GH) deficiency or multiple pituitary hormone deficiencies. Dehydroepiandrosterone sulfate, dehydroepiandrosterone, delta 4-androstenedione and testosterone were measured by RIA in 339 blood samples collected during the study period. Mean plasma androgen levels are normal in isolated GH deficiency. Patients with multiple pituitary hormone deficiencies, but normal ACTH reserve, have mean levels lower than normal. Patients with multiple deficiencies including ACTH deficiency have still lower plasma androgen levels. Longitudinal analysis of the data, however, shows that patients with either isolated GH deficiency or multiple pituitary deficiencies without ACTH deficiency constitute a heterogeneous population, with either normal or low to very low plasma androgen levels. Treatment with human GH as such does not have any effect on the adrenal androgen secretion. A dissociation is found in some patients between adrenarche and gonadarche, which indicates that the two events are not controlled by the same mechanisms. Our results support the existence of a specific hypothalamic pituitary adrenal androgen-stimulating hormone (AASH). ACTH, although not identical to AASH, is essential for normal adrenarche. Induced puberty with estrogens in girls does not influence plasma androgen levels, and pubic and axillary hair growth is not achieved. It is suggested that replacement treatment with dehydroepiandrosterone sulfate should be administered to girls with hypopituitarism and very low plasma androgen levels at the time of the induction of puberty. Finally, it appears from this study that, to interpret the plasma androgens in hypopituitarism, body surface is as good as bone age. PMID- 6286442 TI - Heterozygotes and cryptic patients in families of patients with congenital adrenal hyperplasia (21-hydroxylase deficiency). HLA and glyoxalase I typing and hormonal studies. AB - In a group of 18 unrelated Danish children with 21-hydroxylase deficiency (21-OH def.), human leukocyte antigen (HLA) typing revealed a significant increase of Bw47 and a significant decrease of B8. HLA studies of the families of 14 probands predicted among the siblings 11 heterozygote carriers and 3 genetically unaffected. Glyoxalase studies showed a recombination fraction of 8%. ACTH stimulated 17-OH progesterone is the only hormone value useful in the discrimination between heterozygotes and normals. Two families are described in detail. In one family, one of two HLA-identical brothers had classical virilizing congenital adrenal hyperplasia (CAH), while the other was a normal boy without 21 OH def. In another family with 3 girls, one had classical, salt-wasting CAH, one had "late onset' CAH, and the third sister and the father shared the HLA-B14 antigen and were shown to have "cryptic' 21-OH def. PMID- 6286444 TI - [Isolation and crystallization of the Clr subunit of the first component of human complement (author's transl)]. AB - Isolation of Clr from Cohn-Fraction I of human plasma is accomplished by a series of steps which include affinity chromatography on IgG-p azobenzyloxyethylsulfonylethoxy-Sepharose CL-4B, ion exchange chromatography and preparative zone electrophoresis. The protein is easily soluble in 0.1 M Tris/HCl buffer, pH 10.0 and is crystallized readily by means of dialysis against 0.15 M saline, pH 7.0 in the form of hexagonal dipyramides. The structural elucidation using X-ray analysis is now possible. PMID- 6286443 TI - Ectopic production of ACTH by Wilms' tumor. AB - The authors present the 2nd documented case of Wilms' tumor associated with the "ectopic ACTH syndrome'. This is a 7 1/2-year-old girl who, on examination at the time of admission, had the classical cushingoid appearance. A large hard mass was palpable in the right side of the abdomen. Hormonal assays were consistent with Cushing's syndrome; the serum ACTH levels were extremely high. After surgical removal of the mass, we suspected a stage I Wilms' tumor; this was confirmed by histopathological studies. After surgery, the girl quickly lost her cushingoid appearance and weight excess. Postoperative serum ACTH levels were normal. Ectopic hormone syndromes associated with tumors in childhood are discussed as well as the possible mechanism involved in the ectopic production of ACTH. PMID- 6286446 TI - Formation of ether lipids from isomeric cis-octadecen-1-ols in normal and neoplastic cells: substrate specificity of enzymes with regard to position of double bonds. AB - The pattern of incorporation of isomeric cis-[1-14C]octadecen-1-ols into alkyl moieties of glycerophospholipids of rat brain as well as of L 1210 and S 180 murine ascites cells revealed the following: 1) The enzymes involved in the biosynthesis of acylalkylglycerophospholamines have no substrate specificity with regard to position of the double bond of either cis-oectadecen-1-ols or of intermediate ether lipids derived therefrom. 2) CDPcholine: diradylglycerol cholinephosphotransferases exhibit a strong preference for acylalkylglycerols with cis-8, cis-9 and cis-10-octadecenyl moieties as substrates. PMID- 6286445 TI - Direct reciprocal allosteric interaction of oxygen and hydrogen carbonate sequence of the haemoglobins of the Caiman (Caiman crocodylus), the Nile crocodile (Crocodylus niloticus) and the Mississippi crocodile (Alligator mississippiensis). PMID- 6286447 TI - Striking structural similarity of pig pancreatic kallikrein, mouse nerve growth factor gamma-subunit and rat tonin. PMID- 6286448 TI - Studies on enzymatic methods for processing hybrid proteins produced by recombinant DNA technology. PMID- 6286449 TI - Mitochondrial formation of OH Radicals by an ubisemiquinone-dependent reaction an alternative pathway to the iron-catalysed Haber-Weiss cycle. PMID- 6286450 TI - Budget, policy changes: NIMH in transition. PMID- 6286451 TI - Cholera and cell regulation. AB - The cholera vibrio, unlike most pathogens, neither invades the body's cells nor causes them direct physical damage and thus does not engender inflammation or fever. Rather, the organism attaches itself to the intestinal epithelium, where it secretes a toxin that deranges cellular function. Study of the toxin's structure and mechanism of action is helping to elucidate normal enzymatic processes. PMID- 6286452 TI - Lung infiltrates following removal of a neck mass. PMID- 6286453 TI - Persistent pneumonia and cough in an afebrile woman. PMID- 6286454 TI - NMR imaging: its potential clinical impact. AB - Nuclear magnetic resonance, a developing technique for visualizing soft tissue pathology that is inaccessible to any modality utilizing ionizing radiation, seems likely to eventually outdistance CT scanning in clinical usefulness. In the near future, NMR, now in its infancy, should complement CT scanning and be increasingly used in probing hematologic and metabolic processes. PMID- 6286455 TI - Amenorrhea, depression and hypercortisolism. PMID- 6286456 TI - Concomitant herpes-monilial esophagitis: case report with ultrastructural study. AB - Herpesvirus and Candida albicans are each well-known pathogens associated with esophagitis, and concomitant infections by both agents are occasionally observed. The case of a 58-year-old man who had been treated for carcinoma of the tonsil and died of confluent bronchopneumonia is presented. Autopsy revealed an esophagitis in which cytologic changes of viral infection were seen in the intact esophageal epithelium along with pseudomycelia of Candida albicans within the ulcer bed. In addition, ultrastructural study showed dual infection by Candida and herpesvirus within individual esophageal epithelial cells at the ulcer edge, a unique demonstration of coexistent intracellular infection. PMID- 6286458 TI - Viruses in the pathogenesis of cervical neoplasia: an update. PMID- 6286457 TI - Asteroid bodies in fibrocystic mastopathy. PMID- 6286459 TI - Immunohistochemical investigations of tumors of supposed fibroblastic-histiocytic origin. AB - The aim of this study was to localize alpha 1-antitrypsin, ferritin, and lysozyme by means of the indirect immunoperoxidase technique and to evaluate the significance of these antigens as markers of histiocytic differentiation in tumors of a supposed dual fibroblastic-histiocytic origin. The series comprised 31 malignant fibrous histiocytomas (MFH) of the pleomorphic, spindle cell, and myxoid types, four cutaneous fibrous histiocytomas, and four atypical fibroxanthomas, four dermatofibrosarcoma protuberans, and two osteoclastomas of bone. For comparison, 15 soft tissue sarcomas of various other types were examined. Of the MFHs of the pleomorphic type, 18 of 22 (82 per cent) were positively stained for alpha 1-antitrypsin and 12 of 22 (54 per cent) were positively stained for ferritin. Of the five MFHs of the spindle cell type, none was positively stained for alpha 1-antitrypsin, three were positive for ferritin, and one was positive for lysozyme. None of the myxoid variants (corresponding to grade I-II myxofibrosarcoma) was positively stained for either of the antigens. These results and the observations made on the cutaneous fibrous histiocytomas, atypical fibroxanthomas, dermatofibrosarcoma protuberans, and the various soft tissue sarcomas indicated that 1) alpha 1-antitrypsin is a valuable marker of histiocytic differentiation in both benign and malignant fibrous histiocytomas, 2) ferritin can be visualized in more than half of these fibroblastic-histiocytic tumors, and the presence of ferritin distinguishes the spindle cells of these tumors from fibroblasts of connective tissue and most fibrosarcomas, and 3) lysozyme, although a good marker of histiocytic differentiation in ordinary histiocytes and benign fibrous histiocytomas, is a poor marker of neoplastic histiocytes of malignant tumors. The results further support the concept that MFH is a tumor of a dual fibroblastic-histiocytic origin. PMID- 6286460 TI - A glomus tumor of the jejunum masquerading as a carcinoid tumor. PMID- 6286461 TI - The Mendelian inheritance of a human X chromosome-specific DNA sequence polymorphism and its use in linkage studies of genetic disease. AB - A recombinant DNA sequence, lambda RB6, was isolated from a human X chromosome library and shown to be X-specific by hybridisation to DNA from a human-mouse somatic cell hybrid containing X as the only human chromosome. The cloned sequence was located on the long arm distal to Xq13 using a human-mouse somatic cell hybrid containing a partial human X chromosome. DNA samples isolated from control human females were digested with the restriction enzyme MspI, and analysed by "blotting" and hybridisation to the radioactive cloned DNA. Eight of 14 individuals from a random population showed a single hybridising band 7.5 kilobase pairs (kb) in length, but six showed an additional band 10.1 kb in length. DNA from 12 members of a family with X-linked thyroxine-binding globulin deficiency was analysed for the segregation of this polymorphism. The results show that the polymorphism is inherited in a Mendelian fashion, and that the disease locus is not closely linked to the polymorphic site. Such polymorphisms will be useful as markers for chromosome mapping and for the antenatal diagnosis of genetic diseases. PMID- 6286462 TI - Genetic aspects of nonchromaffin paraganglioma. PMID- 6286464 TI - A comparison of some biochemical properties of liver thiosulphate sulphurtransferase from guinea pig (Lepus caniculus) & albino rat. PMID- 6286463 TI - Effect of prolonged antiorthostatic hypokinesia on adrenocortical and renal function in healthy individuals. PMID- 6286465 TI - Effect of some radiosensitizing drugs on human erythrocyte membrane--spin label study. PMID- 6286466 TI - Coloured derivatives of plastoquinone-A & dibromothymoquinone: possible role in photosynthesis. PMID- 6286467 TI - Macrophage migration inhibition as a correlate of cell-mediated immunity to herpes simplex virus type 2 in mice. AB - Mice inoculated intraperitoneally or intravenously with herpes simplex virus type 2 (HSV-2) develop a focal necrotizing hepatitis. The livers show expanding foci of necrosis and increasing virus content during the first days of the infection with maximal titers achieved on day 3. The clearance of virus from the organ is manifest from day 4 onward with the most dramatic fall in virus content occurring between days 4 and 5. The development of immunity during the course of infection was assessed by adoptive transfer experiments and by measuring macrophage migration inhibition factor (MIF) production of spleen cells in an indirect agarose microdroplet assay. Antiviral activity of adoptively transferred spleen cells was demonstrable from day 4 of the infection when 50 X 10(6) spleen cells were transferred into recipient mice infected 24 h previously. MIF production in spleen cell cultures stimulated with antigen was found to be specific in that activity was only detected in cultures derived from immune mice and stimulated with the virus antigen. The response was found to be antigen-dose and cell-number dependent. Significant MIF production was demonstrable in spleen cell cultures derived from mice 3 days after the infection, i.e. concomitant with the initiation of recovery and before antiviral activity can be detected in transfer experiments. It is suggested that a delayed type hypersensitivity reaction with lymphokine production leading to recruitment of macrophages and their retention and activation in the foci of infection may be a major factor in the recovery from the infection. PMID- 6286468 TI - Immunoglobulin secretion by human B lymphocytes exposed to herpes simplex type I antigen. AB - Herpes simplex type I (HSV I) antigen was studied for its capacity to induce immunoglobulin(Ig)-secreting cells in human peripheral-blood lymphocytes. Ig secreting cells were detected using an indirect hemolytic plaque assay. These results were compared to those obtained by using pokeweed mitogen (T-dependent) and Epstein-Barr virus (T-independent) to induce Ig-secreting cells. The HSV I induction of Ig synthesis was T-dependent, similar to the pokeweed-mitogen system. Peripheral-blood mononuclear cells depleted of T cells did not produce immunoglobulin. In contrast, the same T-depleted cell populations stimulated with Epstein-Barr virus (another member of the Herpes group) produced Ig. Lymphocytes in the HSV-I-stimulated cultures were found to secrete immunoglobulins of the IgA, IgG, and IgM classes. It was the lymphocytes from seropositive individuals that made Ig after HSV I stimulation. Such responses suggest that this system may be antigen-specific. This T-dependent virally induced Ig synthesis system may be useful in studying immunodeficient and immunocompromised individuals. PMID- 6286469 TI - Mitogen-induced changes in glycolytic enzymes of mouse lymphocytes: influence of insulin on cell activation in vitro. AB - Activated mouse spleen lymphocytes have increased amounts of the glycolytic enzymes pyruvate kinase and lactate dehydrogenase. No changes were found in hexokinase and in the gluconeogenic enzyme fructose 1,6-diphosphatase. Concanavalin A-activated T cells give higher activities of those enzymes than lipopolysaccharide-activated B lymphoblasts. Insulin treatment results in a stronger increment of the enzyme activity of mitogen-activated cells. Insulin inhibits the initial proliferation induced by either Con A or LPS, but a 50% increase in antibody-forming cells was found in LPS-treated cultures. Insulin may favour the differentiation of activated cells by increasing the rate of the glycolytic pathway. PMID- 6286471 TI - Radioimmunoassay for major external glycoprotein of murine mammary tumor virus. PMID- 6286472 TI - Effect of T & B cell immunosuppression on the release of Marek's disease virus from the feather follicles of chickens. PMID- 6286470 TI - Enhanced production of IgE and IgG antibodies associated with a diet enriched in eicosapentaenoic acid. PMID- 6286473 TI - Coronavirus in respiratory infection. PMID- 6286474 TI - Prevalence of antibodies against herpes simplex and adenovirus in oral and cervical cancer patients--a preliminary report. PMID- 6286475 TI - Surgical management of unilateral proptosis. PMID- 6286476 TI - Effect of AV3V lesions on development of DOCA-salt hypertension and vascular Na+ pump activity. AB - We studied the effects of anteroventral third ventricle (AV3V) lesions on the vascular Na+-pump activity of deoxycorticosterone acetate-salt (DOCA-salt) treated rats. Blood pressures and Na+-pump activity of the isolated tail arteries, measured as ouabain-sensitive 86Rb-uptake, were determined in untreated control rats, DOCA-salt treated rats, rats with AV3V lesions, and rats with AV3V lesions which were treated with DOCA-salt. Control rats receiving DOCA treatment developed higher blood pressures than rats receiving no DOCA treatment. Placement of AV3V lesions prior to administration of DOCA prevented the increase in blood pressure. Vascular Na+-pump activity in the DOCA-treated group was reduced by 20% compared to all other groups. The AV3V lesions prevented the suppression of Na+ pump activity caused by DOCA treatment. Suppression of vascular Na+-pump activity was due to a humoral substance since Na+-pump activity of tail arteries from control rats incubated in plasma from DOCA-salt treated rats was suppressed by 25% when compared to those incubated in control plasma. Our findings support the hypothesis that a circulating pressor substance is at least partially responsible for the development of DOCA-salt hypertension and that the mechanism by which AV3V lesions prevent DOCA hypertension may be through the interruption of secretion, transport, or synthesis of this factor. PMID- 6286477 TI - Modulation of brain angiotensin-converting enzyme by dietary sodium and chronic intravenous and intracerebroventricular fusion of angiotensin II. AB - Angiotensin-converting enzyme (ACE) in rat brain closely resembled that in lung in its kinetics with the substrate Hip-His Leu, the inhibitors SQ 20,881 and SQ 14,225, and iun its Cl- activation profile. Modification of dietary NaCl intake was associated with marked changes in brain ACE activity. Sodium-loaded rats had lower activity of ACE in hypothalamus, striatum, and midbrain, and higher activity in spinal cord compared to controls. In sodium-restricted rats, ACE was elevated in pituitary and depressed in spinal cord. Chronic intravenous infusion of angiotensin (AII) was associated with a pattern of changes partly resembling sodium loading: ACE was depressed in hypothalamus and striatum but elevated in midbrain. After chronic intracerebroventricular infusion of AII, ACE was elevated in striatum and hippocampus, and depressed in spinal cord; a pattern of changes quite different from those associated with intravenous AII. These results show that ACE in several brain regions is sensitive to dietary sodium intake and support the hypothesis that angiotensin-containing neurons in these areas might be responsive to NaCl status of the animal. The observed changes in brain ACE do not seem to be explained in any simple manner by changes in circulating or central angiotensin II. PMID- 6286478 TI - Potassium relaxation of vascular smooth muscle from DOCA hypertensive pigs. AB - This study was designed to characterize potassium-induced relaxation in vascular smooth muscle during the development of deoxycorticosterone acetate (DOCA) hypertension. Pigs were implanted subcutaneously with 100 mg/kg DOCA. Mean arterial pressure in the DOCA-treated pigs reached levels approximately 37% greater than controls. In some pigs, the left hindlimb vascular bed was "protected" from the rise in arterial pressure by ligation of the iliac artery. Arterial strips from DOCA hypertensive and normotensive pigs relaxed in response to potassium after contraction induced by norepinephrine in potassium-free solution. Arterial strips from DOCA hypertensive pigs showed greater relaxation than did those from normotensive pigs. The magnitude of relaxation in femoral arteries from "protected" hindlimbs was similar to that in arteries from the contralateral unoccluded limb. Potassium-induced relaxation in tail arteries from DOCA hypertensive pigs was more sensitive to ouabain inhibition than that from normotensive pigs. Relaxation induced by potassium varied with: 1) length of incubation in potassium-free solution; 2) concentration of added potassium; and 3) concentration of norepinephrine added during the potassium-free interval. The amplitude of potassium-induced relaxation is believed to be a functional index of the activity of the electrogenic sodium-potassium transport system. These experiments support the hypothesis that vascular smooth muscle from DOCA hypertensive animals has increased electrogenic sodium pump activity. The development of this vascular change parallels the increase in blood pressure induced by mineralocorticoid excess. PMID- 6286479 TI - Aldosterone binding sites in aortic cell cultures from spontaneously hypertensive rats. AB - Spontaneously hypertensive rats and rats made hypertensive by deoxycorticosterone salt treatment have in common increased Na+ and K+ permeability and transport in their aortic cells. These changes may be important factors in the development of the hypertensive state and may be mediated by mineralocorticoid binding to intracellular sites in the aorta. Therefore, we examined 3H-aldosterone binding in aortic cell cultures from spontaneously hypertensive rats and normotensive Wistar-Kyoto rats. Vascular corticoid binding sites in the two strains were compared by Scatchard analysis of Kd and Bmax, pH and temperature stability, and subcellular binding. By all of these criteria normotensive rats. These results indicate that the underlying genetic defect in spontaneous hypertension is not an intrinsic cellular defect which alters mineralocorticoid binding in the aorta. PMID- 6286481 TI - Methionine enkephalinergic neuronal activity in cerebral nuclei of spontaneously hypertensive rats. AB - The role of methionine enkephalin (ME) neurons in the development of genetic hypertension in SHR is the subject of this study. Methionine enkephalin-like immunoreactivity (MELI) and ME receptor binding (MERB) levels were assayed quantitatively by microdensitometry of fluorescence micrographs and autoradiographs of 85 cerebral nuclei and areas of both young and adult spontaneously hypertensive rats (SHR). Normotensive Wistar Kyoto rats (WKY) were used as controls. In young SHR, both MELI and MERB levels were markedly higher in the n. dorsalis nervi vagi, n. amygdaloideus medialis, and group of stria terminal nuclei than in those of young WKY, while both levels were lower in the n. reticularis lateralis, n. corporis mamillaris lateralis, and n. arcuatus. MELI levels in the tractus spinalis nervi trigemini and MERB in the n. tractus spinalis nervi trigemini and median eminence were also lower in young SHR, whereas MERB in the n. amygdaloideus centralis was higher. Alteration in these nuclei was no longer detectable in adult SHR. Whereas in adult SHR, both MELI and MERB levels in the n. reticularis medialis were higher than those of adult WKY, and MELI in the n. accumbens septi and MERB in the n. caudatus were also higher, while MELI in the area lateralis hypothalami was lower than that in adult WKY. The findings indicate that activation of ME neurons in the n. dorsalis nervi vagi and limbic area and also a decrease in ME neuronal activity in the area spinalis nervi trigemini, n. reticularis lateralis, and n. arcuatus may be casually related to the development of hypertension and hyperreactivity in SHR. PMID- 6286480 TI - Chronic effects of ACTH and cortisol excess on arterial pressure in normotensive and hypertensive dogs. PMID- 6286482 TI - Mechanisms of hypertension in the overweight. PMID- 6286483 TI - Physiologic regulation of arterial pressure. An overview. AB - Research of the past 30 years has produced information showing the close interrelationships of neural mechanisms, the renin-angiotensin-aldosterone system, and salt balance as determinants of arterial pressure, both normal and elevated. Contemporary emphases are on central sites of neural regulation of arterial pressure, and an interesting sidelight of the conventional approaches is the possibility that the endorphin-enkephalin system may have a role in hypertension. Salt balance is critically important for people with salt-dependent hypertension; mechanisms of this dependency have not been defined but possible candidates are activation of the sympathetic nervous system, release of a natriuretic factor that causes vasoconstriction as well as salt loss, and renal neural interactions. PMID- 6286485 TI - [3 tumors of the cheek]. PMID- 6286484 TI - Influence of cations on kinetics of angiotensin II binding to adrenal, renal, and smooth muscle receptors. AB - A number of biological responses to angiotensin II have been demonstrated to be modulated acutely by cations, but the exact mechanism has not been elucidated. We have utilized a radioreceptor assay for angiotensin II to determine whether this acute regulatory mechanism could be related to a change in either number or affinity of angiotensin II binding to receptors. Three target tissues were used (adrenal glomerulosa, glomeruli, and uterine smooth muscle). Both mono- and divalent cations influence the kinetics of angiotensin II binding in a similar manner in all tissues. Divalent cations increase the number of receptor sites to a greater extent than did monovalent cations, while monovalent cations changed binding affinity to a greater extent than did divalent cations. These studies demonstrate that both the number and affinity to a greater extent than did monovalent cations, while monovalent cations changed binding affinity to a greater extent than did divalent cations. These studies demonstrate that both the number and affinity of angiotensin receptors can be rapidly modulated by a variety of cations. PMID- 6286486 TI - Rescue and serotypic characterization of noncultivable human rotavirus by gene reassortment. AB - Thirty-three of 50 noncultivable human rotavirus strains from a variety of locations were successfully rescued by gene reassortment. The serotype of each of the 33 strains was investigated by a qualitative cytopathic effect neutralization assay. Nineteen strains resembled the previously characterized human rotavirus serotype Wa, whereas three strains were serologically related to the DS-1 strain. Eleven strains appeared to be serotypically distinct from the Wa and DS-1 strains and thus apparently represent one or more new human rotavirus serotypes. PMID- 6286487 TI - Definition of human rotavirus serotypes by plaque reduction assay. AB - Twenty different human rotavirus reassortants were characterized serologically by a plaque reduction assay as belonging to one of three distinct serotypes. Fourteen were similar if not identical to our prototype Wa strain; two were like the prototype DS-1 strain, and four belonged to a third serotype for which a prototype has not yet been selected. Hyperimmune sera raised against the three serotypes were required to distinguish among them, since postinfection sera had lower titers and were more cross-reactive than hyperimmune sera. These results confirmed the ability of a qualitative cytopathic neutralization test to predict correctly the Wa or DS-1 serotype. A strain of rhesus rotavirus (MMU 18006) was identified as belonging to the newly defined third serotype. Finally, an attempt was made to correlate previously published serotype analysis by neutralization of fluorescent cell-forming units with the results determined by the plaque reduction neutralization assay. PMID- 6286489 TI - Defective parvoviruses acquired via the transplacental route protect mice against lethal adenovirus infection. AB - Adeno-associated virus type 1 (AAV-1) interfered with the replication of its murine adenovirus (MAV) helper in primary mouse kidney cells and in 1-day-old ICR mice. Mice carrying AAV-1 acquired via the transplacental route were protected against lethal infection with MAV. The replication of AAV-1 in these mice could be triggered by multiple challenges with MAV, and antibodies to AAV-1 were subsequently detected. PMID- 6286488 TI - Intratypic and intertypic specificity of lymphocytes involved in the recognition of herpes simplex virus glycoproteins. AB - Cytotoxic T lymphocytes (CTL) were generated in C57BL/6 mice with herpes simplex virus type 1 (HSV-1) (strains KOS, 17, HFEM, and mP) and HSV-2 (strains 186, G, and GP6). Effector lymphocytes were tested for cytotoxicity against syngeneic HSV 1- and HSV-2-infected cells in a 5-h 51Cr release assay. HSV-1 strain HFEM was found to induce CTL efficiently only when 100-fold more virus was used as compared with HSV-1 strains KOS, 17, and mP. All HSV-1 and HSV-2 strains induced cross-reactive populations of CTL. CTL generated by HSV-1 KOS and HSV-2 186 also demonstrated cross-reactivity in an ear-swelling model for delayed-type hypersensitivity. Lymphocytes generated by all HSV-2 strains were highly efficient at lysing HSV-1-infected target cells. However, HSV-2-infected target cells were found to be less susceptible to lysis by either HSV-1 or HSV-2 CTL than were HSV-1-infected target cells. The lowered susceptibility of HSV-2 infected cells was not due to an inefficient infection of BL/6 WT-3 cells as measured by standard growth assays and infectious center assays. Varying the multiplicity of infection or the time of infection did not increase the susceptibility of HSV-2-infected target cells to lysis by CTL. Increasing the effector-to-target-cell ratio resulted in an increased lysis of both HSV-1- and HSV-2-infected target cells by CTL, but the level of HSV-2-infected target cell lysis still did not approach the level of HSV-1-infected target cell lysis. HSV-2 infected cells were as efficient as HSV-1-infected cells in the cold cell competition assay employed in reducing the lysis of 51Cr-labeled, HSV-1-infected target cells. In addition, HSV-2-infected cells were susceptible to lysis by HSV immune serum and complement. PMID- 6286490 TI - Coxsackievirus B infection in pregnant mice and transplacental infection of the fetus. AB - Direct instillation of coxsackievirus B1 into the gastrointestinal tracts of albino mice caused viremia in more than 85% of the animals within 1 day. In pregnant mice infected early in gestation (7 days), the geometric mean titer of virus in the blood was lower (P = 0.02) and the duration of viremia was shorter (P = 0.07) than in nonpregnant female mice, but infection of the heart, liver, and uterus did not differ on each of 5 days after infection. Although transplacental infection of the placenta or fetus or both occurred, the high spontaneous abortion rate (48%) obviated comparison of transplacental infection in these mice with mice infected later in gestation. Pregnant mice infected in the third trimester had significantly greater geometric mean titers of virus in the blood, heart, liver, and uterus, and infection persisted longer than in nonpregnant mice (P = 0.04). A very high geometric mean titer of virus was recovered from the uteri of these mice for 3 days after infection, whereas simultaneous geometric mean titers of virus in the placentas and fetuses were lower. In the majority of third trimester pregnant mice, virus was found in low titers in the fetuses at 2 and 3 days after maternal infection, and virus was not detected after day 3. We conclude that coxsackievirus B1 infection in late gestational pregnant mice is more severe than in mice at earlier gestational stages and in nonpregnant mice and that transplacental infection of the fetus occurs transiently during maternal infection. This model will prove useful in the study of perinatal enterovirus infection and in examination of the numerous factors that may influence outcome of infection of perinatally infected newborn infants. PMID- 6286491 TI - Adhesion of Entamoeba histolytica trophozoites to human erythrocytes. AB - To understand the mechanism of Entamoeba histolytica adhesion, we characterized the binding of trophozoites to human erythrocytes (RBC) in suspension by measuring the kinetics of amoeba-RBC complex formation. Adhesion was very efficient, since most of the amoebae were complexed with RBC after only 5 min at 37 degrees C in mixtures containing 10(4) amoebae and 10(6) RBC per ml; the adhesion rate depended on amoeba and RBC concentrations, but not on the A, B, and O human blood groups, and was maximal at 37 degrees C and pH 7.3 in the presence of 4 mM Ca2+ and 1 mM Mg2+. Adhesion was prevented if trophozoites were fixed with glutaraldehyde, but only decreased slightly if RBC were previously fixed; it decreased in the absence of glucose and was inhibited as a function of the concentration of cytochalasin B and of the metabolic inhibitors bathophenanthroline and 8-hydroxyquinoline. From these results we conclude that E. histolytica adhesion is an active process that depends on the amoebal cytoskeleton and metabolic energy and on the mobility of both amoebal and RBC surface ligands. PMID- 6286492 TI - Dietary hepatic cholesterol elevation: effects on coxsackievirus B infection and inflammation. AB - Mice made hypercholesterolemic (HC) by diet are highly susceptible to coxsackievirus (CV) B5, whereas normal adult animals remain resistant. In attempting to define those dietary-induced physiological changes which contribute to altered resistance, a strong association between accumulation of intrahepatic cholesterol and increased CV B5-induced mortality was demonstrated, with maximum susceptibility to CV coinciding with a 2.5-fold increase in the ratio of hepatic cholesterol to protein. This metabolic imbalance was associated with a lower clearance rate of CV from the blood and liver of C57BL/6 mice, although virus specific neutralizing antibody production was unaltered. In addition to CV, HC mice were more susceptible to an intravenous inoculation of Listeria monocytogenes in comparison to controls. The macrophage stimulant Corynebacterium parvum failed to increase resistance of HC mice to a high dose of CV B4 and L. monocytogenes and failed to induce the hepatomegaly, splenomegaly, and cellular infiltrate seen in the liver and spleen of normal animals. Furthermore, the peritoneal monocytic infiltrate induced by thioglycolate in normal animals was absent in HC mice. Results from these experiments suggest that decreased resistance to CV in the HC host is attributed to a defect in the nonspecific immune responses of macrophages and monocytes which are of primary importance in resistance to this virus and other infectious agents. PMID- 6286494 TI - Effect of in vitro preincubation of polymorphonuclear leukocytes on formylmethionyl-leucyl-phenylalanine-induced chemiluminescence. AB - When polymorphonuclear leukocytes (PMNL) and soluble or particulate matter interact, the cells produce chemiluminescence. The chemotactic peptide formylmethionyl-leucyl-phenylalanine induced a two-peak chemiluminescence response in PMNL. The response was modified, both in magnitude and in the time course of the response, when PMNL were incubated at 22 degrees C for 15 to 120 min before the addition of stimulus. The cellular response to formylmethionyl leucyl-phenylalanine was also changed by the addition of cell-free supernatants from stored PMNL suspensions to freshly prepared PMNL before the addition of formylmethionyl-leucyl-phenylalanine. This indicated that PMNL during storage released substances that changed the chemiluminescence response. A similar change could be obtained by addition of purified myeloperoxidase to freshly prepared PMNL before the addition of stimulus, indicating that the myeloperoxidase hydrogen peroxide system can function as a modulator of the cellular response to chemotactic factors. PMID- 6286495 TI - Persistent Varicella-Zoster virus infection in a human rhabdomyosarcoma cell line and recovery of a plaque variant. AB - Varicella-zoster virus (VZV) has been found to persistently infect the human rhabdomyosarcoma cell line A204. Infectious center assays and fluorescent antibody staining demonstrated continuous production of infectious VZV and viral antigen. The level of infection determined by fluorescent antibody staining was variable, and usually only a small percentage of the cells were capable of producing plaques in permissive fibroblasts. The extent of infection was similar in cell cultures passaged at split ratios of 1:2 or 1:10 and grown at 33 or 37 degrees C. VZV recovered from A204 cells several months after establishment of the persistent infection had markedly increased syncytia-forming activity as compared with the parental VZV grown in human diploid fibroblast cells and the three monkey kidney-derived cell lines Vero, CV-1, and MA104. The expression of this altered phenotype continued after serial passage of the cell-associated virus in human diploid fibroblast and Vero cells. Consequently, we designated the reisolated VZV as plaque variant A. The buoyant densities of VZV plaque variant A and VZV DNAs in CsCl gradients were indistinguishable. PMID- 6286493 TI - Gram-positive bacteria-induced granulocytopenia and pulmonary leukostasis in rabbits. AB - Pneumococci have been shown to induce granulocytopenia and pulmonary leukostasis which might contribute to morbidity and mortality in pneumococcal sepsis. We studied whether other gram-positive species (groups A and B streptococci, Staphylococcus aureus, Bacillus cereus, and Clostridium perfringens) could also induce these phenomena. Rabbits were challenged with nonviable preparations of each species, and all five species induced profound granulocytopenia (mean decrease of 90%) and pulmonary leukostasis. In vitro studies of serum incubated with these species of bacteria showed a mean consumption of total hemolytic complement of 90%, a mean increase of chemotactic activity for granulocytes of 204%, and a mean augmentation of granulocyte adherence of 45% (compared with 18% for the control). Infusion of sonicate-exposed sera induced granulocytopenia in recipient rabbits. Thus, several nonviable gram-positive species can interact with serum to activate the complement system, generate C5a bioactivity, augment granulocyte adherence, and generate a neutropenia-inducing factor. These alterations may contribute to granulocytopenia or pulmonary leukostasis, which may play a role in the morbidity and mortality associated with gram-positive bacterial infections. PMID- 6286496 TI - Herpes simplex virus-specific serum immunoglobulin a: detection in patients with primary or recurrent herpes infections and in healthy adults. AB - A sensitive radioimmunoassay was used to determine levels of herpes simplex virus (HSV)-specific immunoglobulin A (IgA) in serial serum samples drawn from patients with primary HSV infections and from persons with recurrent HSV infections, and in single samples from 90 healthy adults. Significantly rising HSV IgA titers were detected in patients with primary infections, whereas those with recurrent infections had nonfluctuating titers. Sera of IgG-seropositive healthy adults were all positive for HSV-specific IgA without special pretreatment. PMID- 6286498 TI - Delayed hypersensitivity to murine cytomegalovirus and its depression during pregnancy. AB - A delayed hypersensitivity skin test for murine cytomegalovirus is described, in which ear swelling is measured after injection of heat-killed virus. The response appeared within 4 days of infection, peaked at 8 days, and remained at high levels for at least 100 days. When live virus was inoculated into the ear of previously uninfected mice, a much greater degree of ear swelling was seen, maximal at 7 days, but without growth of virus in the inoculated ear. Mice infected with avirulent (cell culture-passaged) virus gave greater delayed hypersensitivity responses than with those infected with virulent (salivary gland passaged) virus. One of the strains of mice that is susceptible to murine cytomegalovirus (C57BL) developed greater delayed hypersensitivity responses than did the resistant strain (C3H). The delayed hypersensitivity response of neonatally infected mice was greatly depressed during pregnancy and lactation. PMID- 6286497 TI - Inhibition of herpes simplex virus multiplication by activated macrophages: a role for arginase? AB - Proteose-peptone-activated mouse macrophages can prevent productive infection by herpes simplex virus in neighboring cells in vitro whether or not those cells belong to the same animal species. The effect does not require contact between the macrophages and the infected cells, may be prevented by adding extra arginine to the medium, and may be reversed when extra arginine is added 24 h after the macrophages. Arginase activity was found both intracellularly and released from the macrophages. The extracellular enzyme is quite stable; 64% activity was found after 48 h of incubation at 37 degrees C in tissue culture medium. No evidence was found that the inefficiency of virus replication in macrophages was due to self-starvation by arginase. As might be predicted macrophages can, by the same mechanism, limit productive infection by vaccinia virus. PMID- 6286499 TI - Non-specific influence of antibiotics on the course of infectious processes. AB - The clinical significance of a concomitant, non-specific influence of antibiotics on immune defence mechanisms was studied by evaluating the death rate in mice experimentally infected with highly resistant or primarily resistant microorganisms. It could be shown that the mortality rate of mice infected with Enterobacter cloacae or Candida albicans significantly increased under treatment with cefoxitin, whereas treatment with cefotaxine or lamoxactam either had no effect, or even resulted in a better survival rate in comparison to controls. These results run parallel to an inhibition (cefoxitin) or stimulation (cefotaxime and lamoxactam) of antibody production. The effect of cefoxitin on the course of experimental infections could be compensated for by the concomitant application of sodium-8-chlorotheophyllinate which promotes antibody formation. None of these antibiotics showed any additional effect in animals treated with cyclophosphamide. From these observations it was concluded that the influence of antibiotics on certain immunological parameters assayed in vitro may be reflected in comparable effects on the course of infections in vivo; this implies that under certain clinical conditions, the immunological side-effects of antibiotics may be of practical therapeutic significance. PMID- 6286500 TI - A prospective study on infectious mononucleosis in childhood--symptoms, serology, Epstein-Barr-Virus specific leukocyte migration inhibition. AB - The clinical course, serological changes and the development of the specific cell mediated immune response to Epstein-Barr-Virus (EBV), measured in terms of leukocyte migration inhibition, were followed in 40 children suffering from an EBV infection. The patients were followed for between six and 24 months. Although the majority of the children were under six years of age, they presented a typical clinical course; heterophil antibodies could only be demonstrated in 60% of the cases. Anti-VAC-IgM and IgG antibodies were found in all patients during the acute phase, but no anti-EBNA could be demonstrated. In children under three years of age, no antibodies against the D component of the early antigen were found; this antibody was found in 50% of the adolescents. An antibody against the R component of the early antigen could be demonstrated in 73% of the children five to six months after the onset of the disease. Specific leukocyte migration inhibition was present only during convalescence or later. A relationship between the appearance of anti-EBNA and the development of specific leukocyte migration inhibition has been established. PMID- 6286501 TI - Pivmecillinam in complicated urinary infections failing to respond to conventional therapy. AB - Forty-three patients with a history of recurrent urinary infections, many of whom had renal involvement and had recently failed treatment, were then treated with pivmecillinam (400 mg 8-hourly for seven days). One week after the end of treatment the bacteriological cure rate was 72.1%. During the following four weeks, however, several of the cured patients relapsed, so that the cure rate five weeks after the end of treatment was only 46%. Thus, it is recommended that such patients should begin prophylactic treatment if found to be abacteriuric after the one week follow-up. Pivmecillinam was extremely well tolerated. PMID- 6286502 TI - [Therapy with cefotaxime-cefotaxime/ticarcillin for bronchopulmonary infections in patients under intensive care. (author's transl)]. AB - 55 intensive care patients with an internal underlying disease were treated with cefotaxime, and 12 patients with bronchopulmonary infections were treated with the combination cefotaxime/ticarcillin. The following aspects were evaluated: clinical success, antimicrobial activity of bacteria in evidence before and after therapy, persistence of pathogens, and resistance. Renal function was monitored continuously in 21 patients over a period of 20 days in order to demonstrate nephrotoxic side-effects of cefotaxime when administered simultaneously with furosemide (in doses of mre than 1 g/day). In view of the clinical results and of the fact that the cefotaxime/ticarcillin combination is well tolerated by the kidneys, the possibility of efficient chemotherapy without aminoglycosides is being discussed for treatment of bronchopulmonary infections in patients under intensive care. PMID- 6286503 TI - [Antibiotic resistance of anaerobic bacteria (author's transl)]. AB - Reference data on the overall resistance patterns of anaerobic bacteria are needed since in individual clinical cases susceptibility testing is complicated by the long generation time and difficulties of test conditions. Benzylpenicillin and tetracyclines are the drugs of choice against Clostridium perfringens. Newer cephalosporins are also active, whereas other Clostridium species have been found resistant. Beta-lactam antibiotics show high activity against Peptococcaceae; resistance to clindamycin should be noted, however. The good in vitro activity of tetracyclines, clindamycin and erythromycin against Propionibacterium acnes could be confirmed in the systemic and local therapy of acne vulgaris. In contrast to gram-positive anaerobic bacteria, only some penicillin and cephalosporin derivatives are sufficiently active against gram-negative rods of the Bacteroides group. Sporadic resistance is seen to the newer tetrayclines, nitroimidazole compounds and to lincomycins. Nevertheless, these substances must be considered as standard therapeutics for infections caused by anaerobic gram-negative bacteria. It should be mentioned that cefoxitin may offer new therapeutic possibilities. PMID- 6286504 TI - Diarrhoea in foals. PMID- 6286505 TI - Effect of Bordetella pertussis on alpha 1 and beta-adrenergic and cholinergic muscarinic receptors in guinea pig lung membranes. AB - After intraperitoneal injection of Bordetella pertussis vaccine to guinea pigs, the alpha 1- and beta-adrenergic and cholinergic muscarinic receptors of the whole lung were measured by binding assays with the radioisotope-labeled antagonists, 3H-prazosin, 1-(3)H-dihydroalprenolol and 1-(3)H-quinuclidinyl benzilate, respectively. Injection of guinea pigs with pertussis vaccine resulted in an increase in the maximum binding (Bmax) of 3H-prazosin, while there was about a 30% reduction in 1-(3)H-dihydroalprenolol binding to beta-adrenergic receptors. No difference was observed in the dissociation constant (KD) for binding of each ligand to alpha 1- and beta-adrenergic receptors. 1-(3)H quinuclidinyl benzilate binding indicated that the Bmax and KD for cholinergic muscarinic receptor in pertussis-sensitized guinea pigs were not significantly different from those normal control animals. PMID- 6286506 TI - Diurnal variation of gonadotrophin receptors in the rat testis. AB - Groups of 6 adult male rats were killed at 4-h intervals over 24 h periods. The serum samples were assayed for LH, FSH, prolactin (Prl) and testosterone (T). LH, FSH and lactogen receptors were measured in the testis homogenates. In the 2 experiments performed, statistically significant (P less than 0.01-0.05) positive correlation was observed between serum LH and T levels, as well as between serum LH and testis LH receptor concentrations. Significant (P less than 0.05) diurnal variation was seen in peripheral serum T (high levels at 08.00-12.00 h, low levels at 16.00-20.00 h) and LH (high at 24.00-08.00 h, low at 12.00-20.00 h), whereas FSH and Prl levels did not display significant variations over the period of 24 h. The testicular content of LH receptors showed significant (P less than 0.05) diurnal periodicity in the 2 experiments performed. Maximal binding measured in each experiment was between 04.00 and 08.00 h. The low binding was in one experiment between 12.00 and 20.00 h, and in the other between 20.00 and 24.00 h. No consistent variation was measured in testicular FSH or lactogen receptors. These results suggest that, in addition to differential stimulation of steroidogenesis, physiological variations in gonadotrophin levels can induce autoregulatory changes in the testicular gonadotrophin receptors. PMID- 6286507 TI - Human testicular LH receptors: correlations with circulating gonadotrophins and testicular steroid secretion. AB - Testicular androgen production is regulated by circulating LH, the effect of which is mediated by its testicular membrane receptors. In the present study, testicular [125I]hCG binding in man was investigated and found to be characterized by saturability and high affinity (mean KD in testes of 21 patients with prostatic carcinoma = 1.64 X 10(-10) M), and stimulated testosterone production in dispersed interstitial cells in vitro. The concentrations of the LH receptors correlated with serum FSH concentrations (r = 0.52, P less than 0.05) but not with circulating LH levels. Neither had they any correlation with intratesticular testosterone, 5 alpha-dihydrotestosterone, androstenedione, progesterone, 17-hydroxyprogesterone or pregnenolone concentrations, which may be due to the fact that LH receptors are confined to Leydig cells, whereas steroids may be unevenly distributed in the different cells of the testis. In contrast, the concentrations of the LH receptor displayed positive correlations with the concentrations of testosterone (r = 0.81, P less than 0.001), androstenedione (r = 0.54, P less than 0.05), 17-hydroxyprogesterone (r = 0.76, P less than 0.001) and progesterone (r = 0.60, P less than 0.05) in the spermatic vein serum of the patients. Our data suggest that the Leydig cells mainly responsible for steroid secretion into the blood are under gonadotrophic control exerted via their receptors, whereas Leydig cell function is not rapidly reflected in steroid concentrations within the testis itself. PMID- 6286508 TI - Direct inhibition of testicular steroidogenesis and gonadotrophin receptor levels by [(imBzl)-D-His6, Pro9-NEt]GnRH and [D-Trp6, Pro9-NEt]GnRH, potent agonists of GnRH. AB - The effects of [(imBzl)-D-His6, Pro9-NEt]GnRH and [D-Trp6, Pro9-NEt]GnRH on testicular function in rats was evaluated. In adult rats the administration of 0.01, 0.1 or 10 micrograms of either agonist induced rapid increases in serum LH, FSH and testosterone (T) levels which started to decline within several hours. Both agonists caused a decrease in testicular LH and FSH receptor concentrations. The testicular FSH receptor concentration started to decline earlier than the LH receptor concentration but, both reached their lowest levels by day 2 after the administration of the agonists. The recovery of FSH receptor content was slower than that of LH. The extrapituitary effects of the 2 agonists were investigated in immature hypophysectomized animals. Administration of hCG (5 IU daily) to hypophysectomized rats for 5 days caused an increase in serum T levels. Concomitant administration of either of the agonists (10 micrograms) inhibited the steroidogenic action of hCG. Administration of the agonists alone caused a reduction in testicular LH receptor concentration in hypophysectomized rats. Treatment of the hypophysectomized rats for 0-4 days suggested that the direct antitesticular action of the agonists requires 1-2 days to become evident. PMID- 6286509 TI - The deleterious effect of mechanical dissociation of rat testes on the functional activity and purification of Leydig cells using Percoll gradients. AB - The effect of mechanically dispersing rat testes prior to collagenase treatment on the activity of the subsequently isolated Leydig cells has been investigated. The latter were obtained by centrifugation of the testes cells on Percoll density gradients. It was found that the lutropin-stimulated testosterone and cyclic AMP production were 5 and 3 times higher, respectively, in the purified Leydig cells when the mechanical dispersion was omitted, whereas the specific binding of human choriogonadotrophin was unchanged. It is concluded that mechanical treatment of rat testes, unlike mouse testes, decrease the activity of the Leydig cells. PMID- 6286510 TI - Reaction of 99mTc-diphosphonate with N-donor ligands. PMID- 6286511 TI - Synthesis of 18F-6-fluoropurine and 18F-6-fluoro-9-beta-d-ribofuranosylpurine. AB - Introduction of fluorine-18 into the 6-position of purines is described. 18F-6 fluoropurine and 18F-6-fluoro-9-beta-D-ribofuranosylpurine were prepared with high radiochemical yields by nucleophilic displacement of the trimethylammonio group of purine with 18F-fluoride under mild conditions. 18F-labeling conditions such as reaction temperature and time, addition of crown ether and dose of a substrate have been studied. Under adequate conditions, yields of about 38% in the case of free base and about 63% in the case of ribofuranosyl derivative have been obtained. The latter compound has also been prepared in a non-carrier-added state. The stability of 18F-purines was examined in Tris buffer at 37 degrees. PMID- 6286512 TI - Non-invasive assessment of regional myocardial function at rest and during exercise after aorto-coronary bypass operation. PMID- 6286513 TI - Valve replacement for chronic mitral insufficiency; long-term follow up using technetium pertechnetate scintigraphy. AB - Technetium Pertechnetate Scintigraphy was performed in 12 patients (9 men and 3 women, average age 48 +/- 13 years) on an average 19.8 +/- 11 months after valve replacement for chronic mitral insufficiency. Preoperative conventional ventriculography had revealed increased enddiastolic and endsystolic volumes (230.9 +/- 66 resp. 73.5 +/- 26 ml). 59.4 +/- 9% of the stroke volume (157.5 +/- 47 ml) regurgitated into the left atrium. Ejection-fraction (67.7 +/- 6%) was normal in all cases. Postoperative first pass technique was employed using a fast multicrystal camera (Baird Atomic, system 77). Volumes were assessed measuring normalized total count rate (count integral of the left ventricle divided by the maximum count density). 19.8 +/- 11 months after valve replacement enddiastolic and endsystolic volumes revealed normal at rest (121.8 +/- 38 resp. 52.8 +/- 35 ml) and after maximum exercise (122.0 +/- 29 resp. 37.6 +/- 22 ml). Ejection fraction responded physiologically to exercise by rising from 57.7 +/- 12 to 69.7 +/- 10%. PMID- 6286514 TI - A monoclonal antibody with anti-Burkitt lymphoma specificity. I. Analysis of human haematopoietic and lymphoid cell lines. AB - 38-13 is a hybridoma-produced monoclonal rat IgM which appears to define a Burkitt's lymphoma-associated antigen (BLA). In this paper, we described the reactivity of 38-13 with a panel of human haematopoietic and lymphoid cell lines. In indirect immunofluorescence (IF) assays, 15 of 26 Burkitt's lymphoma (BL) lines studied were clearly stained with 38-13 (from 13 to 100% positive cells) by microscope, with varying numbers of heavily labelled cells. In these positive cell lines, fluorescence-activated cell-sorter (FACS) analysis demonstrated that BLA was actually present on all the cells. Positive BL included Epstein-Barr virus (EBV) genome-carrying lines and EBV-negative ones; thus, BLA is not related to the presence of EBV. Most of the 15 BL cells that reacted with 38-13 contained a typical t(8;14) translocation, but had variant translocations such as t(2;8) and t(8;22). The cells were derived from BL patients of different geographical origins and clinical features. Four BL lines were poorly stained and seven were negative with 38-13 in IF assays. The 32 EBV-positive lymphoblastoid cell-lines (LCL) studied were negative. In three line pairs, consisting of a tumor line and an LCL from the same patient, only the BL line was demonstrated to react with 38 13. A series of non-BL cells, including haematopoietic, lymphoid and solid tumor lines, all failed to react with 38-13. Various attempts to modulate the expression of BLA on BL cells were unsuccessful. However, it cannot be ruled out that BLA is actually a transient B-cell differentiation marker. PMID- 6286515 TI - The successful treatment of mycotic infections in immunosuppressed renal transplant recipients with ketoconazole. AB - Ketoconazole, a new broad-spectrum antimycotic drug, was administered to six renal transplant recipients with mucocutaneous and/or systemic candidosis. A beneficial clinical and microbiologic effect was seen in the treated patients. This orally administered drug was well tolerated, and side effects were not evident. Our results indicate that good treatment of mycotic infections can be expected even in patients with impaired graft function, since ketoconazole metabolism occurs mainly in the liver. PMID- 6286516 TI - Correction of leg inequality in the Klippel-Trenaunay-Weber syndrome. AB - Eight patients with Klippel-Trenaunay-Weber syndrome were treated for leg length inequality of as much as 10 cm in one ten year old, and one thirteen year old. In most cases adequate correction was achieved with epiphyseal stapling during active growth or definitive epiphysiodesis in older children. Bone shortening procedures in full-grown individuals may be indicated but the basic condition may make healing difficult. Ligation of the femoral artery had no effect on the growth of the affected leg. PMID- 6286517 TI - Independence of physical working capacity from increased glucocorticoid level during short-term exercises. AB - To investigate the influence of glucocorticoid levels on the physical working capacity, 18 trained and 18 untrained male subjects were examined before and after a short-term graded exercise test. By administration of dexamethasone during 3 days prior to the test or by injecting 25 U of corticotropin 30 min pre exercise, the blood cortisol response to exercise was altered. It was found that the physical working capacity, as evaluated by PWC170, maximal oxygen deficit, and maximal pedaling rate were not influenced by the glucocorticoid response to the test. It is suggested that the effects of metabolic processes induced by adrenocortical response to exercise are seen only after a lag period. PMID- 6286518 TI - Compulsory admissions to mental hospitals in six countries. PMID- 6286520 TI - Adrenal responsiveness in children with steroid responsive idiopathic nephrotic syndrome. AB - Impaired adrenal cortical responsiveness to ACTH has been reported in children with steroid responsive nephrotic syndrome. Plasma cortisol levels at 8 AM and 2 hours post ACTH stimulation were measured on 42 occasions in 33 children with steroid-responsive nephrotic syndrome. Twenty-two of 42 fasting 8 AM plasma cortisol and 39 of 42 post-ACTH plasma cortisol values were subnormal. Plasma cortisol values were similar in children evaluated prior to, immediately following or 2-24 moths after prednisone therapy. No relationship was observed between 8 AM or post-ACTH plasma cortisol values and serum albumin concentration or the presence of edema. Impaired adrenal responsiveness to ACTH was not helpful in predicting the length of clinical remission. Hydrocortisone replacement therapy given to five children with impaired ACTH-responsiveness did not alter the rate of relapses observed prior to this treatment. PMID- 6286519 TI - The melanotic neuroectodermal tumor of infancy. Report of two cases and review of the literature. AB - The melanotic neuroectodermal tumor of infancy (MNTI) is a rare childhood neoplasm with an alarming but classical clinical presentation. It appears as a rapidly enlarging mass in the jaws of skull of infants and unless MNTI is considered in the differential diagnosis, the lesion can easily be mistaken for a malignant neoplasm. Although possessing an aggressive growth rate and radiographic appearance, the MNTI almost always behaves in a benign fashion and can be treated with local excision. However, recent reports of malignant behavior as well as of occasional recurrences make close follow-up important. Approximately 139 cases of the MNTI have been reviewed and tabulated with respect to age at discovery, sex, tumor location, length of follow-up, and whether recurrence occurred, 2 additional cases, 1 of which was in the mandible, are presented along with theories of origin and recommended therapy. PMID- 6286521 TI - Effect of dilution on thermal neutron induced inactivation of deoxyribonuclease I. PMID- 6286522 TI - In vitro uptake of radiocolloids by rat peritoneal macrophages and human tumor cells. PMID- 6286523 TI - Aortic flux of radiopharmaceuticals in atherosclerotic rabbits. PMID- 6286525 TI - Mechanism of contraction in cardiac muscle. PMID- 6286524 TI - Tumors metastatic to the eye and adnexa. PMID- 6286526 TI - Lithium pharmacokinetics, duration of therapy, and the adenylate cyclase system. AB - The elimination half-life of lithium carbonate in red blood cells, plasma, and urine was measured in 30 patients hospitalized for primary affective disorders. The duration of lithium treatment at the time of sampling was found to have a direct effect on lengthening time-course. Those on their initial course of lithium had the lowest half-lives (in days): 1.12 (urine), 1.28 (plasma), and 1.22 (red blood cells); those with less than 1 year of lithium had intermediate values: 1.85, 1.65, and 1.75, and those with more than 1 year of continuous lithium administration had the highest mean half-lives: 2.40, 2.43, and 2.24. The significance of these results for urine (p less than 0.01) and plasma (p less than 0.05) indicates further evidence that lithium may stimulate the production of an endogenous regulator of lithium efflux. 8 of these patients had lumbar punctures before lithium administration: those with previous treatment with lithium had higher mean cyclic adenosine monophosphate levels than those taking it for the first time. These results are discussed in the context of the possible mechanisms of lithium action. PMID- 6286528 TI - Metabolism of cholecalciferol in vitamin D intoxicated chicks. AB - Vitamin D intoxication was induced in chicks by treatment with large amounts of radioactive cholecalciferol (vitamin D3) either by s.c. injections or by stomach tube. Hypercalcemia and nephrocalcinosis were present, confirming toxicity. The distribution of cholecalciferol and its metabolites in the tissues of the intoxicated birds was compared with that in birds that were treated with physiological amounts of radioactive cholecalciferol. Treatment with pharmacological doses resulted in marked elevation of cholecalciferol and its metabolites in all tissues examined, including elevation of 1 alpha,25 dihydroxycholecalciferol in the intestine. The predominant form of cholecalciferol in these birds was found to be the unchanged vitamin, whereas in birds treated with physiological doses 25-hydroxycholecalciferol was the predominant metabolite. The route of vitamin administration was found to be of importance only when pharmacological doses were given: generally, higher levels were noted when administered via s.c. injections than via stomach tube, except in the arteries. It is suggested that in vitamin D intoxication, the factor responsible for the pathological changes in soft tissues is cholecalciferol itself. High levels of 1 alpha,25-dihydroxycholecalciferol may be responsible for the hypercalcemia. PMID- 6286527 TI - Thyroid pertechnetate scan in neonatal hyperthyroidism. PMID- 6286529 TI - Parotitis and pancreatitis complicating infectious mononucleosis. PMID- 6286530 TI - Clinical features of viral and bacterial gastroenteritis in hospitalized children. AB - The clinical course of 119 children, hospitalized with acute gastroenteritis of known etiology, was studied. The severity of disease was measured by a scoring system that showed that the disease caused by Escherichia coli was the most severe, followed in decreasing order by Salmonella, human reovirus-like agent (HRLA) and Shigella. The clinical features of salmonella and shigella infections differed from those reported in other countries. Fifty percent of all patients recovered after 5 days of treatment, but 20% of the patients with bacterial infections continued to have symptoms for 10 days or longer. The relationships between etiological agents and various symptoms were measured by odds ratios. These associations may provide a basis for clinical prediction of the etiological agent. PMID- 6286531 TI - alpha 1-antitrypsin deficiency in Israeli children: a five-year survey. AB - In order to elucidate the importance of alpha 1-antitrypsin (AAT) deficiency as a cause of liver disease in children in Israel, we conducted a retrospective 5-yr study. The screening of 51 liver biopsies for the presence of AAT inclusion bodies was performed using the immunoperoxidase technique. Serum AAT concentrations of 300 pediatric patients determined during the same period were reviewed. No case of AAT deficiency was detected. We conclude that AAT deficiency is rare in Israel. PMID- 6286532 TI - The laboratory approach to the diagnosis of infectious mononucleosis. PMID- 6286533 TI - In-vitro-induced human T cells cytotoxic against herpes-simplex-virus-infected targets: modulation by theophylline of the induction and effector stages. AB - We have shown in previous studies that the cytotoxic effector cells in in-vitro induced cytotoxicity against herpes-simplex-virus-infected targets are principally T gamma cells and that they derive from theophylline-resistant T mu cells. In the present work, we studied the effect of theophylline on the induction and expression of cytotoxic activity in T-cell subsets. When T cells were prefractionated before the induction phase, the theophylline-resistant (Thr) subset, as determined by E-rosetting, was unaffected by the presence of theophylline during sensitization, whereas T mu and unfractionated T cells were inhibited. On the other hand, when theophylline was present during the cytotoxicity assay, all T-cell subsets were similarly affected. Studies of kinetics revealed that a short preincubation with the drug had an enhancing rather than an inhibitory effect on both the proportions of T gamma and Thr and on their cytotoxic activity at 3 days. PMID- 6286534 TI - [Multiple atypical milker's nodes in scalded areas]. AB - This case report describes the occurrence of multiple atypical milker's nodules in scalded skin. This hitherto unknown manifestation of paravaccinia virus infection is transmitted indirectly by virus contaminated water. PMID- 6286535 TI - Environmental radon daughter alpha dose factors in a five-lobed human lung. AB - The alpha dose from 222Rn daughters per unit exposure (rad per WLM or rad per year per pCi 222Rn/m3) to basal cells in bronchial epithelium in a five-lobed human male lung is calculated. These factors are then derived for women, children and infants by scaling the dimensions of the male lung. The 222Rn daughter characteristics chosen are those typical of environmental atmospheres. Both active and resting breathing patterns are utilized in the calculations to yield the overall dose per unit exposure in typical population exposures. PMID- 6286537 TI - An optimized scheme for measurement of the concentrations of the decay products of radon and thoron. AB - Radon daughter measurement methods have not previously been fully optimized in the presence of thoron daughters, the presence of which leads to an increase in the total potential alpha energy. Taking into account the fluctuations associated with flow rate and concentration as well as the statistics of decay, optimized counting schemes were determined for the daughters of radon and thoron for different sampling times over a wide range of counting intervals. The counting schemes examined were optimized for maximum precision. Counting schemes corresponding to impractically long counting intervals, and counting times which required measurements beyond an 8 hr working shift were rejected. Under these constraints the most appropriate sampling time was found to be 10 min. PMID- 6286536 TI - An evaluation of unattached radon (and thoron) daughter measurement techniques. AB - The collection efficiencies of parallel plate, inertial impactor and wire screen devices reported in the literature for measurement of the unattached fraction of radon (or thoron) daughters are calculated for collection of the attached fraction of the aerosol, using real uranium mine aerosol activity-size distributions. The attached activity collection efficiencies may be as much as 4.3%, which, given the typically low values of the unattached fraction (a few per cent), results in a very substantial error in the measurement of the latter. Published unattached fraction measurements in the environment may therefore be high and should be interpreted with care. PMID- 6286538 TI - Localization of SCEs and their possible relationship to dA . dT- or dG . dC clusters, respectively, in Chinese hamster V79-E chromosomes. PMID- 6286539 TI - Local authority psychiatric hostel catering and the health of residents--a case report. PMID- 6286540 TI - Breast-milk intake measurement in mixed-fed infants by administration of deuterium oxide to their mothers. AB - Breast-milk intakes in infants can be estimated from measurements of water turnover rates after the oral administration of small doses of deuterium oxide (2H2O) to the infant or its mother. However, when the former method is used it is not possible to distinguish water intakes from milk and those from other sources. Giving a 2H2O dose to the mother avoids this problem. The theoretical basis for these techniques is examined and useful compromises between the requirements of theory and practice are suggested. Results obtained for breast-milk intakes of Gambian and Papua New Guinean children are used to illustrate the potential value of the procedures. PMID- 6286541 TI - Heterogeneity of concanavalin A binding by mouse peritoneal macrophages. AB - The binding of the lectin concanavalin A (Con A) to the cell surface of monocytes and macrophages collected from the stimulated peritoneal cavity of mice was investigated electron microscopically with horseradish peroxidase-gold as an indirect marker. Individual cells were identified by the cytochemical localization of peroxidatic (PO) activity. In monocytes and monocyte-derived macrophages with PO activity in cytoplasmic granules, the degree of Con A binding was lower than in resident macrophages with PO activity in the rough endoplasmic reticulum and nuclear envelope. An even higher degree of Con A labelling was found on the surface of cells devoid of PO activity. Since the above-mentioned cell populations show a different degree of lectin binding, it is suggested that lectin labelling methods might offer a new tool for quantitative investigation of the differentiation of monocytes and resident macrophages. PMID- 6286542 TI - Enzymatic cleavage prior to antibody incubation as a method for neuropeptide immunocytochemistry. AB - When deplasticized Epon sections were treated with endo- and/or exopeptidases prior to incubation with antibodies, the neuropeptide immuno-reactivity of secretory nerves was often altered in a predictable way. Cleavage of neurosecretory material in octopus nerves by trypsin and carboxypeptidase-B enhanced enkephalin-like immunoreactivity, while Molluscan neuropeptide-like immunoreactivity was prevented by tryptic cleavage. The enzyme effects indicated the occurrence of a heptapeptide (Tyr-Gly-Gly-Phe-Met/Leu-Arg-Phe) that contains both the enkephalin and the Molluscan neuropeptide sequence. Vasopressin terminals of the rat neurohypophysis, which presumably contain enkephalin precursor sequences, exhibited enkephalin-like immunostaining after tryptic cleavage. ACTH/beta-endorphin cells of the rat intermediate pituitary, which synthesize the enkephalin sequence at the N-terminus of Beta-endorphin, exhibited enkephalin=like immunoreactivity when sections were treated with alpha chymotrypsin or trypsin, but not after incubation with leucine-aminopeptidase or carboxypeptidase-B. Enkephalin-like immunostaining could not be induced in any way in ACTH/beta-endorphin cells of the anterior pituitary. Enzymatic cleavage may give additional information in immunocytochemical localization studies on neuropeptide sequences in secretory nerves and hormonal granules. PMID- 6286543 TI - Glioblastoma multiforme: treatment by large dose fraction irradiation and metronidazole. AB - In an attempt to overcome the possible radioresistance of glioblastoma multiforme related to the large shoulder on the in vitro survival curves and to sensitize hypoxic tumor cells, a treatment protocol was instituted at Yale University Medical Center and affiliated hospitals, using large dose fraction irradiation therapy in conjunction with the hypoxic cell sensitizer metronidazole. Nineteen patients with biopsy-confirmed, previously untreated, cerebral grade IV glioblastoma multiforme were, following surgery, irradiated once a week at 600 rad per fraction, 3.5 to 4 hours after ingestion of metronidazole, 6 gm/m2. A total of 7 treatments were employed, with all patients maintained on antiseizure medications and corticosteroids. Metronidazole levels were determined prior to each treatment and patients were followed closely clinically and with serial computerized tomography (CT) scans. The treatment was well tolerated, in general, with no untoward side effects related to the high dose fraction irradiation. The majority of the patients experienced varying degrees of gastrointestinal upset lasting up to several hours following metronidazole administration. Three patients died of pulmonary emboli. One patient experienced moderately severe ototoxicity. A median survival of 9.4 months was obtained for all 19 patients, suggestive of a prolongation of survival compared to historical controls treated with conventionally fractionated radiation or with unconventional radiation fractionation schemes and metronidazole or misonidazole. PMID- 6286544 TI - Protective drugs in cancer therapy: optimal clinical testing and future directions. PMID- 6286545 TI - Circumvention of the tumor membrane barrier to WR-2721 absorption by reduction of drug hydrophilicity. AB - In attempting to account for the ability of most solid tumors to restrict the absorption of WR-2721, aminopropyl-aminoethylphosphorothioate, we examined a number of drug characteristics which might allow for this restriction, and observed that drug hydrophilicity was a major contributing factor. When the highly hydrophilic WR-2721 was dephosphorylated, the drug became less hydrophilic and could readily cross tumor cell membranes. In addition, conventional radioprotectants, such as cysteine and mercaptoethylamine, were shown to be less hydrophilic than WR-2721 and also to cross tumor membranes readily. Therefore, drug hydrophilicity would appear to be the factor underlying the ability of WR 2721 to selectively protect normal tissues while most other protectors alter the radiation resistance of normal and tumor tissue alike. A red blood cell model for studying this problem in greater detail is described. PMID- 6286546 TI - In vitro studies on the absorption of WR-2721 by tumors and normal tissues. PMID- 6286547 TI - Experimental radiotherapy with WR-2721 and misonidazole. AB - The data from a large series of experiments on mouse skin and tumors are summarized. Radioprotection with WR-2721 has been observed in both tumors and normal tissues. The protection factors are generally, but not always, higher for skin than for tumors. The protection observed in skin is greater for mice irradiated breathing air than for those irradiated in oxygen. It is postulated that the different protection factors observed in different normal tissues and tumors may reflect differences in tissue oxygenation levels. The combination of misonidazole and WR-2721 has been studied in terms of the modification of radiosensitivity and also as the modification of lethal toxicity. An interaction has been observed in all aspects. The toxicity of WR-2721 increases in the presence of misonidazole. The WR-2721 radioprotection of both skin and tumors decreases if the sensitizer is added. Likewise the radiosensitization with misonidazole is diminished when WR-2721 is present. These results indicate an interaction at the site of radiation injury and they also demonstrate that WR 2721 can adequately penetrate into hypoxic tumor cells. PMID- 6286548 TI - Radioprotection of mouse skin by WR-2721: the critical influence of oxygen tension. AB - The epidermal clone assay has been used to study the radioprotective effect of WR 2721 on mouse skin under different conditions of oxygenation and under anoxia. The skin has shown a progressive decrease in sensitivity as the inspired gas was changed from 100% oxygen towards 0% oxygen. Compared with mice breathing 100% oxygen, those breathing air are partially protected. The inspired oxygen concentration to given half the full oxygen effect is 10--12%. The radioprotection observed with 400 mg/kg WR-2721 is markedly dependent on the ambient oxygen concentration. The protection factor is 1.1 or less in mice breathing 5%, 1% of 0% oxygen. Protection is maximal (1.95) in air and in 50% oxygen and diminishes to 1.6 at higher oxygen tensions. PMID- 6286549 TI - Protective effects of WR-2721 against radiation-induced injury of murine gut, testis, lung, and lung tumor nodules. AB - WR-2721 (S-2-(3 aminopropylamino) ethylphosphorothioic acid) has been investigated for its ability to protect gut, lung, and testis, as well as fibrosarcoma (FSa) tumor nodules, in the lungs of mice from gamma-radiation injury. This compound greatly protected jejunum and testis epithelial cells. FSa micrometastases in the lung were protected to a lesser extent than jejunum and testis. Conversely, WR-2721 was not able to protect the lung against radiation induced enhancement of tumor metastases formation generated by intravenously injected FSa cells. PMID- 6286550 TI - Influence of WR 2721 on the efficacy of radiotherapy and chemotherapy of murine tumors. AB - The effect of WR 2721 on the response of tumors to radiation, antineoplastic alkylating drugs, and DNA binding agents was evaluated and compared to the degree of normal tissue protection provided by WR 2721 against these agents. WR 2721 administered to mice bearing P388 leukemia or Lewis lung carcinoma was found to reduce the radiosensitivity of the leukemia and lung tumor by dose modifying factors of 1.4 and 1.3, respectively. WR 2721 protected bone marrow, intestine, and skin from radiation by factors of 1.9, 1.5, and 1.8. WR 2721 protected mice from the lethality of cyclophosphamide by a factor of only 1.2 whereas protection from melphalan toxicity was more dramatic with a dose modifying factor of 1.6. In chemotherapy studies of established M5076 ovarian tumor, the combination of WR 2721 plus cyclophosphamide was equivalent in activity to cyclophosphamide alone. WR 2721 did not modify the antitumor activity of melphalan in early Lewis lung carcinoma but did decrease the antileukemic effects of this agent by a factor of 2.6 indicating tumor protection greater than host protection in the leukemia. The antitumor activity of the DNA binding agents etoposide (VP16--213) and mitoxantrone against systemic P388 leukemia was not diminished by WR 2721, while a substantial increase in host toxicity was noted for the combinations. The protective effects of WR 2721 against radiation and drug damage were, therefore, not entirely selective for normal tissues. In some cases the degree of tumor protection can be similar to, or greater than, normal tissue protection. PMID- 6286551 TI - Effects of WR 2721 on cyclic nucleotide levels and lysosomal enzyme activities. AB - The radioprotectant WR 2721 administered intraperitoneally to rats at 30--40 min before exposure to 1000 rad of 60Co caused a reduction in spleen lysosomal enzyme activity and fluctuations in cyclic nucleotide levels. Liver and spleen cAMP levels from drug-treated animals were higher at 3--6 hr postirradiation whereas spleen cGMP levels were lower at 1--4 hr postirradiation than irradiated controls not given the radioprotectant. At 3 days after radiation exposure, spleen cyclic nucleotide levels and lysosomal enzyme activities in rats given WR 2721 were closer to the level of nonirradiated control rats than of irradiated rats receiving saline. PMID- 6286552 TI - Sodium hydrogen-S-(3-amino-2-hydroxypropyl) phosphorothioate (WR-77913): toxicity and bone marrow radioprotection. AB - Experiments have been carried out to compare the toxicity and radioprotective effect of sodium hydrogen-S-(3-amino-2-hydroxypropyl) phosphorothioate (WR-77913) with those of S-2-(3-aminopropylamino)ethyl phosphorothioic acid (WR-2721). The drugs were given intraperitoneally to 12 week-old female BALB/c mice 30 minutes before whole body irradiation. Lethality at 30 days was the endpoint used. The drug LD50/30 was 678 mg/kg for WR-2721 and 3574 mg/kg for WR-77913. The LD50/30 for WR-77913 combined with 500 mg/kg of WR-2721 was 3328 mg/kg. The LD50/30 for misonidazole was 380 mg/kg when given in combination with 500 mg/kg of WR-2721 and 801 mg/kg when combined with 2200 mg/kg of WR-77913. Protection of bone marrow by WR-77913 and WR-2721 was comparable at doses close to the maximum tolerable dose (MTD, drug dose lethal to 10% of the animals at 30 days), but WR 77913 gave better protection at 35% of the MTD. These characteristics of low toxicity, non-additive toxicity with WR-2721, less toxicity in combination with misonidazole and adequate bone marrow protection at 25% of the MTD, make WR-77913 a protector worthy of further investigation. PMID- 6286553 TI - Combined radiosensitization and radioprotection for oral cavity tumors: study with an oral cavity tumor model. AB - Adjuvant therapy with combined radioprotectors and radiosensitizers has not been extensively examined. We investigated the use of combined S-2-(3 aminopropylamino) ethylphosphorothioic acid (WR-2721), misonidazole (Ro-07-0582) and single dose radiotherapy in treating the EMT 6/Ky tumor implanted in the buccal mucosa of Balb/c mice. Our results demonstrate that single dose irradiation plus WR-2721 or misonidazole prolonged the mean survival time (MST) of EMT 6/Ky bearing Balb/c mice significantly compared to either controls or single dose irradiation plus WR-2721 and misonidazole. PMID- 6286554 TI - Clinical trials of WR-2721 with radiation therapy. AB - The radioprotector with clinical potential, S-2-(3 aminopropylamino) ethylphosphorothioic acid (WR-2721) is undergoing two Phase I trials. The objectives of these trials are 1) to determine the maximum tolerated dose (MTD) of WR-2721 in a single dose and 2) to determine the highest dose of WR-2721 that can be tolerated daily in the greatest number of fractions per week. A total of 65 patients have been treated. The single maximum tolerated dose has not yet been reached, though 740 mg/m2 is well tolerated. A single dose of 910 mg/m2 has been successfully administered to one patient. The multiple dose MTD is at an early stage with patients currently receiving 170 mg/m2 four times a week. Among the toxicities noted in both trials are hypotension, hypertension, emesis and somnolence. In addition, in the multiple dose trial there have been three patients who have had allergic reactions including one which was life threatening. Phase II studies are planned and will begin when the maximum tolerated dose is established from each Phase I trial. PMID- 6286555 TI - WR-2721: enhanced chemoprotection by increasing cyclophosphamide activation with phenobarbitol. PMID- 6286556 TI - Phase I clinical trials of WR-2721 with alkylating agent chemotherapy. PMID- 6286557 TI - The time course of radioprotection by WR 2721 in mouse skin. AB - The radioprotective effect of 400 mg/kg of WR 2721 on mouse skin has been investigated over a series of times from 5 to 60 minutes after intravenous or intraperitoneal injection of the drug. The acute desquamation reaction on the hind leg of white mice was studied. Dose response curves were obtained for the average reaction over 15 to 25 days after single-dose irradiation. A high dose rate electron beam (13--17 Gy/min) was used to minimize the irradiation time. Single doses of 20 to 60 grays were given. Dose modifying factors (DMFs) of 1.7- 2.1 were obtained at 30 to 60 minutes, but only 1.1 to 1.3 at 5 minutes and intermediate values at 10 and 15 minutes. DMFs rose slightly faster and to slightly higher values after i.v. than after i.p. injection of WR 2721. We conclude that 30 minutes is the shortest interval which should be used between injection of Wr 2721 and irradiation with this normal tissue. PMID- 6286558 TI - High-dose radiation therapy with low-dose (pulsed) BCNU in malignant gliomas: an Eastern Cooperative Oncology Group (ECOG) report. AB - Based on preliminary favorable results with the use of postoperative high-dose radiation therapy for malignant gliomas and encouraging experimental studies which indicated that fractionated does of BCNU given 16 hours before irradiation significantly enhanced the effect of radiation both in vitro and in vivo, the Eastern Cooperative Oncology Group (ECOG) decided to explore the use of high-dose radiation therapy in combination with pulsed low doses of Group (ECOG) decided to explore the use of high-dose radiation therapy in combination with pulsed low doses of BCNU in a high-risk Phase I-II pilot study. The radiation therapy consisted of delivering 6000 rad to the whole brain plus a boost of 1000 rad to the tumor. BCNU was given i.v. 16 hours before the next radiation fraction with doses of 20 mg/M2 twice weekly for six weeks beginning on the second day of irradiation. Twelve patients were treated (5 with Grade III and 7 with Grade IV Astrocytomas). The median survival for the entire group was 80 weeks (30 weeks for Grade IV and 210 weeks for Grade III tumors). These results are not difficult from what was achieved by high-dose radiation alone. Subsequent experimental data seem to indicate that probably the smallest dose of BCNU that should be used in pulsed BCNU combined modality therapy is 80 mg/M2. In general, the treatment as offered in this study was well tolerated. There were no fatal or life-threatening hematologic toxicities and only one patient exhibited a transitory leukopenia of 1600. PMID- 6286559 TI - Enhanced radiation reaction following combination chemotherapy for small cell carcinoma of the lung, possibly secondary to VP16-213. PMID- 6286560 TI - Improved quality of life of patients with small-cell carcinoma of the lung by elective irradiation of the brain. AB - About 30% of all patients with small cell undifferentiated carcinoma of the lung will ultimately develop metastases in the brain. It is now well established that elective whole brain irradiation can prevent such metastases, although it does not increase survival time. However, we have found that such treatment does increase the quality of life for these patients. Those with limited disease can survive with a Karnofsky scale of more than 60 for a mean time of 10 months compared with 6 months for an equivalent group treated only when these brain metastases occur. We have also found that treating patients therapeutically does not insure success as 6/16 eventually relapsed in the brain. We conclude that elective brain irradiation in patients with small cell undifferentiated carcinoma of the lung who present with a limited disease is of value because the quality of life is improved significantly. PMID- 6286562 TI - Increasing spatial awareness in undergraduate nursing students: a viable concept. PMID- 6286561 TI - Fetal lung in organ culture. IV. Supra-additive hormone interactions. AB - Corticosteroids, thyroid hormones, and theophylline have previously been shown to accelerate fetal lung maturation. We have examined the interactions between these agents in relation to phospholipid synthesis in explants of 18-day fetal rat lung in organ culture. Maximal stimulation of the rate of incorporation of choline into phosphatidylcholine, the most abundant phospholipid in pulmonary surfactant, was observed at a dexamethasone concentration of 100 nM. Exposure to 100 nM dexamethasone, 1.0 mM theophylline, or a combination of the two agents for 48 h resulted, respectively, in 144, 157, and 508% stimulation of the rate of incorporation of choline into disaturated phosphatidylcholine. Similar supra additive interactions between dexamethasone and dibutyryl adenosine 3',5'-cyclic monophosphate (cAMP) were observed, but the effects with caffeine were less striking. The increase in the rate of precursor incorporation was associated with a significant increase in the disaturated phosphatidylcholine content of the cultures. Combination of dexamethasone with 100 nM triiodothyronine (the concn producing maximal effects) also resulted in supra-additive stimulation but to a smaller degree. These findings of interactions in vitro suggest that the agents act on the lung at different biochemical sites, but the mechanisms whereby they interact at the cellular level have yet to be established. The data provide a rationale for in vivo animal studies of the effects of combined hormone administration on fetal lung maturation. PMID- 6286564 TI - Teaching psychiatric/mental health nursing via the contract for learning activities. PMID- 6286563 TI - One approach to teaching cultural similarities and differences. PMID- 6286565 TI - Burnout in nursing education. PMID- 6286566 TI - Faculty development in nursing education: a teaching consultation service. PMID- 6286567 TI - Too much instruction--too little learning: an experience in teaching community health nursing. PMID- 6286568 TI - Development and validation of a trigger tape on therapeutic communication. PMID- 6286569 TI - Hospital social workers and nurses: interprofessional perceptions and experiences. PMID- 6286570 TI - Strategies for faculty: teaching the RN student in a BSN program. PMID- 6286571 TI - Strategies for dealing with students whose clinical performance is unsatisfactory. PMID- 6286573 TI - Teaching biomedical science. PMID- 6286572 TI - Development and implementation of a contract grading system. PMID- 6286574 TI - Moral development of health care professionals begins with sensitizing: thirty three sample encounters. PMID- 6286575 TI - Female gender as a variable in the educational process: a review. PMID- 6286576 TI - Moral development: a prerequisite. PMID- 6286577 TI - Geriatric and gerontology nursing curricular trends. AB - The amount of geriatric and gerontology content in a nursing curriculum does not appear to influence students' attitudes about aging. However, based on the findings of this study, specific content areas should be addressed in nursing curricula. These content areas include: 1) social changes as well as physical changes that occur to elderly persons and adaptation patterns of these individuals; 2) usefulness of aged persons and the contributions they can and do make to society; 3) characteristics of the well elderly as opposed to total concentration on the sick elderly. The inclusion of these content areas might assist the nursing student in developing a more positive attitude about aging. PMID- 6286578 TI - Associate degree nursing: implications for the 1980s and beyond. PMID- 6286579 TI - Student grade appeals- procedure and process. PMID- 6286580 TI - Health fair: providing a learning experience through a community service project. PMID- 6286581 TI - Future trends in post-basic nursing education. PMID- 6286582 TI - Insulin-secreting islet cell tumors: establishing a diagnosis and the clinical course for 25 dogs. AB - Twenty-five dogs with insulin-secreting neoplasms of the pancreas were studied. The diagnosis in each case was determined by histologic evaluation of pancreatic tissue obtained at surgery. The breed distribution revealed that German Shepherd Dogs, Irish Setters, and Collies were most commonly represented. Physical examination, complete blood counts, serum biochemical analysis, and urinalysis were of little diagnostic value, aside from the finding of hypoglycemia in 21 of 25 dogs. Radiographs of the thorax and abdomen were noncontributory to the ultimate diagnosis. Prior to surgery, fasting immunoreactive insulin concentrations and blood glucose concentrations were studied. Insulin:glucose ratios, glucose:insulin ratios, and amended insulin:glucose ratios were determined from the insulin and glucose concentrations in a single blood sample in each of 28 trials. In addition, glucagon tolerance tests were performed on 12 dogs. The amended insulin:glucose ratios proved to be the most reliable for diagnosis. Pancreatic masses were evident at surgery in 23 of 25 dogs; the remaining 2 dogs had microscopic evidence of an islet cell tumor. Nineteen of the islet cell tumors were carcinomas and 6 were simply described as "islet cell tumors." The mean life expectancy after surgery was 12.3 months. Treatment for malignant islet cell tumours included frequent feeding glucocorticoids, and diazoxide. PMID- 6286583 TI - Distal polyneuropathy after canine heartworm disease therapy complicated by disseminated intravascular coagulation. AB - A 3-year-old male Setter-type dog had a progressive distal sensorimotor polyneuropathy characterized by weakness, bilateral atrophy of distal appendicular musculature, and reduced response to tactile stimuli. The diagnosis of a distal axonopathy was supported by electromyographic findings of fibrillation potentials, positive sharp waves in distal limb muscles, and absence of evoked action potentials, myopathic changes of atrophic angular fibers, and myelinated nerve fiber depletion in distal parts of peripheral nerves. The neuropathy appeared 5 weeks after 38 days of heparin therapy for disseminated intravascular coagulation. The disseminated intravascular coagulation, a complication of thiacetarsamide therapy for heartworm disease, had resolved 40 days after the end of heparin therapy. The cause of the neuropathy was not determined. PMID- 6286585 TI - Effect of furosemide upon endolymph potassium concentration. AB - Chinchillas were anesthetized with ketamine (40 mg/kg i.m.) and endocochlear potential (EP) and potassium concentration in endolymph (Ke+) were determined in control animals and in animals injected with various doses of furosemide (25, 50 or 100 mg/kg i.v.) by means of microelectrodes inserted into scala media. Control EP and Ke+ in the chinchilla were 81.3 +/- 3.8 mV and 158.5 +/- 3.2 mequiv./l, respectively. Following injection of furosemide, a dose-related fall in EP and Ke+ was observed. However, the EP declined much more rapidly than the Ke+, and recovered more quickly than the latter. The recovery of Ke+ tended to lag behind the EP recovery. The debate over whether potassium transport into endolymph and endocochlear potential generation are related or independent events is discussed in the light of recent literature and the present study. PMID- 6286584 TI - Changes in craniofacial suture metabolism in rats fed a low calcium and vitamin D deficient diet. PMID- 6286586 TI - Pituitary content of gonadotropins and GnRH-receptors in pregnant, postpartum and steroid-treated OVX ewes. PMID- 6286587 TI - A three-way crossover study to compare the pharmacokinetics and acceptability of sultamicillin at two dose levels with that of ampicillin. PMID- 6286588 TI - Transport and catabolism of D-mannose in Rhizobium meliloti. AB - Rhizobium meliloti L5-30 grows on D-mannose as the sole carbon source. The catabolic pathway of D-mannose was characterized. The following activities were present: mannose transport system, mannokinase, and mannosephosphate isomerase. Several mannose-negative mutants were selected; they were classified into three functional groups: group I, mannokinase and mannosephosphate isomerase defective: group II, mannokinase defective; and group III, mannosephosphate isomerase defective. Mannose uptake was an active process, since it was inhibited by azide, dinitrophenol, and cyanide, but not by fluoride or arsenate. Growth on succinate repressed mannose uptake activity. The mannose transport system was present in all the mutants. Uptake studies showed that mannose-negative mutants did not metabolize this sugar. PMID- 6286589 TI - Cointegrate formation between homologous plasmids in Escherichia coli. AB - Conjugation experiments were performed in which the donor was Escherichia coli K 12 strain KP245 containing either R plasmid NR1 plus an ampicillin-resistant derivative of ColE1 (*ColE1::Tn3, called RSF2124) or NR1 plus RSF2124 carrying a cloned EcoRI fragment of NR1. The recipient was the polA amber mutant JG112, in which RSF2124 cannot replicate. Ampicillin-resistant transconjugants can arise only when the genes for ampicillin resistance are linked to NR1 or are transposed to the host chromosome. When EcoRI fragment A of NR1 (20.5 kilobases) was cloned to RSF2124, the frequency of cotransfer of ampicillin resistance with tetracycline resistance was 25 to 60%. Plasmid DNA from these ampicillin resistant transconjugant cells was analyzed by gel electrophoresis and was shown to be a cointegrate of NR1 and the RSF2124 derivative. Analysis of plasmid DNA isolated from donor cultures showed that the cointegrates were present before conjugation, which indicates that the mating does not stimulate cointegrate formation. When the cloned fragment was EcoRI fragment H of NR1 (4.8 kilobases), the frequency of cotransfer of ampicillin resistance with tetracycline resistance was about 4%, and the majority of the ampicillin-resistant transconjugants were found to contain cointegrate plasmids. When the donor contained NR1 and RSF2124, the frequency of cotransfer of ampicillin resistance was less than 0.1%, and analysis of plasmid DNA from the ampicillin-resistant transconjugants showed that Tn3 had been transposed onto NR1. These data suggest that plasmids which share homology may exist in cointegrate form to a high degree within a host cell. PMID- 6286590 TI - Purification and properties of phytate-specific phosphatase from Bacillus subtilis. AB - An enzyme which liberates Pi from myo-inositol hexaphosphate (phytic acid) was shown to be present in culture filtrates of Bacillus subtilis. It was purified until it was homogeneous by ultracentrifugation, but it still showed two isozymes on polyacrylamide gel electrophoresis. The enzyme differed from other previously known phytases in its metal requirement and in its specificity for phytate. It had a specific requirement for Ca2+ for its activity. The enzyme hydrolyzed only phytate and had no action on other phosphate esters tested. This B. subtilis phytase is the only known phytate-specific phosphatase. The products of hydrolysis of phytate by this enzyme were Pi and myo-inositol monophosphate. The enzyme showed optimum activity at pH 7.5. It was inhibited by Ba2+, Sr2+, Hg2+, Cd2+, and borate. Its activity was unaffected by urea, diisopropylfluorophosphate, arsenate, fluoride, mercaptoethanol, trypsin, papain, and elastase. PMID- 6286591 TI - Gratuitous induction by N-acetylmannosamine of germ tube formation and enzymes for N-acetylglucosamine utilization in Candida albicans. AB - N-Acetylmannosamine did not support the growth of Candida albicans, and this sugar was not accumulated by cells. Incubation of starved yeast cells at 37 degrees C with N-acetylmannosamine plus glucose resulted in germ tube formation. Furthermore, N-acetylmannosamine alone induced the uptake system for N acetylglucosamine and the enzymes of the N-acetylglucosamine catabolic pathway to the same extent as the natural substrate. Induction of the uptake system and the enzymes was observed at 28 degrees C without germ tube formation and at 37 degrees C with germ tube formation. N-Acetylmannosamine is thus a gratuitous inducer for enzymes of the N-acetylglucosamine pathway and germ tube formation in C. albicans. PMID- 6286592 TI - Penicillinase-producing Neisseria gonorrhoeae: a molecular comparison of 5.3-kb and 7.4-kb beta-lactamase plasmids. AB - Restriction endonuclease analysis and heteroduplex studies indicate that the only difference between the 5.3-kilobase (kb) and 7.4-kb plasmids from beta-lactamase producing Neisseria gonorrhoeae is that the latter is the 5.3-kb plasmid with a 2.1-kb insertion. The insertion is bounded by inverted repeats of approximately 300 base pairs. Several plasmids from beta-lactamase-producing N. gonorrhoeae isolated at different times and in different countries were compared. Nine 5.3-kb plasmids were examined and found to be indistinguishable, as were sixteen 7.4-kb plasmids. PMID- 6286593 TI - Exonuclease activity from Pseudomonas aeruginosa which is missing in phenotypically restrictionless mutants. AB - A phenotypically restrictionless strain of Pseudomonas aeruginosa was found to lack a deoxyribonuclease specific for linear duplex DNA. The purified enzyme had an optimum pH of 8.5, required MgCl2 (10 mM) for maximum activity, and did not require ATP. Neither the degradation of heat-denatured DNA nor the degradation of bacteriophage F116 DNA was detected. The genome of bacteriophage F116 was shown to possess single-stranded terminal regions, which account for the resistance to degradation and for the ability of the phage to transfect restriction-proficient strains. PMID- 6286594 TI - Mercuric reductase enzyme from a mercury-volatilizing strain of Thiobacillus ferrooxidans. AB - Cell-free mercury volatilization activity (mercuric reductase) was obtained from a mercury-volatilizing Thiobacillus ferrooxidans strain, and the properties of intact-cell and cell-free activities were compared with those determined by plasmid R100 in Escherichia coli. Intact cells of T. ferrooxidans volatilized mercury at pH 2.5, whereas cells of E. coli did not. Cell-free enzyme preparations from both bacteria functioned best at or above neutral pH and not at all at pH 2.5. The T. ferrooxidans mercuric reductase was a soluble enzyme that was dependent upon added NAD(P)H. The enzyme activity was stable at 80 degrees C, required an added thiol compound, and was stimulated by EDTA. Antisera against purified mercuric reductases from transposon Tn501 and plasmid R831 (which inactivated mercuric reductases from a wide range of enteric and pseudomonad strains) did not inactivate the enzyme from T. ferrooxidans. PMID- 6286595 TI - Mapping nonselectable genes of Escherichia coli by using transposon Tn10: location of a gene affecting pyruvate oxidase. AB - Mutants of Escherichia coli K-12 deficient in pyruvate oxidase were isolated by screening for the production of 14CO2 from [1-14C]pyruvate by the method of Tabor et al. (J. Bacteriol. 128:485-486, 1976). One of these lesions (designated poxA) decreased the pyruvate oxidase activity to 10 to 15% of the normal level but grew well. To map this nonselectable mutation, we isolated strains having transposon Tn10 inserted into the chromosome close to the poxA locus and mapped the transposon. These insertions were isolated by the following procedure: (i) pools of Tn10 insertions into the chromosomes of two different Hfr strains were prepared by transposition from a lambda::Tn10 vector; (ii) these Tn10-carrying strains were then mated with a poxA recipient strain, and tetracycline-resistant (Tetr) recombinants were selected; (iii) the Tetr recombinants were then screened for 14CO2 production from [1-14C]pyruvate. This method was shown to give a greater than 40-fold enrichment of insertions of Tn10 near the poxA gene as compared with transduction. Calculations indicate that a similar enrichment should be expected for other genes. The enrichment is due to the much greater map interval over which strong linkage between selected and unselected markers is found in conjugational crosses as compared with transductional crosses. The use of Hfr conjugative transfer allows isolation of transposon insertions closely linked to a nonselectable gene by scoring hundreds rather than thousands of colonies. Using a Tn10 insertion greater than 98% cotransduced with the poxA locus, we mapped the poxA gene on the E. coli genetic map. The poxA locus is located at 94 min, close to the psd locus. The clockwise gene order is ampA, poxA, psd, purA. The poxA mutation is recessive and appears to be a regulatory gene. PMID- 6286596 TI - Regulation of adenylate cyclase synthesis in Escherichia coli: studies with cya lac operon and protein fusion strains. AB - We have isolated cya-lac operon and protein fusions in Escherichia coli K-12, and we used these to study the regulation of cya, the structural gene for adenylate cyclase. Data obtained from these fusion strains suggest that neither cyclic AMP (cAMP) nor the cAMP receptor protein plays a major role in transcriptional or translational regulation of cya expression. Modulation of intracellular cAMP concentrations elicited only weak repression of cya-lac fusion activity under conditions of high intracellular cAMP, relative to fusion activity under conditions of low intracellular cAMP. The functional cAMP receptor protein was required for this effect. Incorporation of delta crp into cya-lac fusion strains did not affect fusion expression in glucose-grown cells as compared with similarly cultured isogenic crp+ strains. Furthermore, 20 independently obtained mutants derived from a cya-lacZ protein fusion strain exhibiting a weak Lac+ phenotype were isolated, and it was determined that the mutants had beta galactosidase activities ranging from 2- to 77-fold greater than those of the parental strain. None of the mutations responsible for this increase in fusion activity map in the crp locus. We used these mutants to aid in the identification of a 160,000-dalton cya-lacZ hybrid protein. Finally, chromosome mobilization experiments, using cya-lac fusion strains, allowed us to infer a clockwise direction of transcription for the cya gene relative to the standard E. coli genetic map. PMID- 6286597 TI - Partial purification and properties of phosphatidylserine synthase from Clostridium perfringens. AB - The membrane-associated phospholipid biosynthetic enzyme cytidine 5'-diphospho 1,2-diacyl-sn-glycerol:L-serine O-phosphatidyltransferase (phosphatidylserine synthase; EC 2.7.8.8) was partially purified 337-fold from a cell-free extract of the gram-positive pathogenic anaerobe Clostridium perfringens (ATCC 3624). The purification procedure included extraction from the cell envelope with the nonionic detergent Triton X-100, followed by affinity chromatography on cytidine 5'-diphosphate-diacylglycerol-Sepharose. When the partially purified enzyme was subjected to polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate, two major bands were evident with apparent minimum molecular weights of 39,000 and 31,000. Activity of phosphatidylserine synthase was dependent on the addition of manganese ions (3 mM) and Triton X-100 (2.7 mM) for maximum activity. The rate of catalysis was maximal at 40 degrees C (with rapid thermal inactivation above this temperature), and the pH optimum was 8.5. The apparent Km values for cytidine 5'-diphosphate-diacylglycerol and L-serine were 0.24 and 0.26 mM, respectively. The synthetic (forward) reaction was favored, as indicated by an equilibrium constant of 82, and the energy of activation was found to be 18 kcal/mol (75,362 J/mol). PMID- 6286598 TI - 2-micrometers DNA-like plasmids in the osmophilic haploid yeast Saccharomyces rouxii. AB - DNA plasmids were detected in two independent strains of Saccharomyces rouxii among 100 yeast strains other than Saccharomyces cerevisiae tested. The plasmids, pSR1 and pSR2, had almost the same mass (approximately 4 X 10(6) daltons) as 2 micrometers DNA of S. cerevisiae. pSR1 and pSR2 gave identical restriction maps with restriction endonucleases BamHI, EcoRI, HincII, HindIII, and XhoI, and both lacked restriction sites for PstI, SalI, and SmaI. These maps, however, differed significantly from that of S. cerevisiae 2-micrometers DNA. Restriction analysis also revealed two isomeric forms of each plasmid and suggested the presence of a pair of inverted repeat sequences in the molecules where intramolecular recombination took place. DNA-DNA hybridization between the pSR1 and pSR2 DNAs indicated significant homology between their base sequences, whereas no homology was detected between pSR1 and pJDB219, a chimeric plasmid constructed from a whole molecule of 2-micrometers DNA, plasmid pMB9, and a 1.2-kilobase DNA fragment of S. cerevisiae bearing the LEU2 gene. A chimeric plasmid constructed with pSR1 and YIp1, the larger EcoRI-SalI fragment of pBR322 ligated with a 6.1 kilobase DNA fragment of S. cerevisiae bearing the HIS3 gene, could replicate autonomously in an S. cerevisiae host and produced isomers, presumably by intramolecular recombination at the inverted repeats. PMID- 6286599 TI - Carbamate kinase from Pseudomonas aeruginosa: purification, characterization, physiological role, and regulation. AB - Pseudomonas aeruginosa PAO1 possessed a carbamate kinase (CKase) distinct from carbamoylphosphate synthetase as well as from a constitutive acetate kinase which also catalyzes the phosphorylation of ADP by carbamoylphosphate. CKase was purified to homogeneity. Polyacrylamide gel electrophoresis of cross-linked CKase in the presence of sodium dodecyl sulfate showed that the enzyme consists of two subunits with identical molecular weights (37,000). The optimal pH of enzyme activity is 7.0. The double-reciprocal plot for carbamoylphosphate was linear at 2 mM ADP, yielding an apparent Km of 5 mM. However, at 0.25 mM ADP, the plot was concave upward, and a Hill plot of the data yielded a coefficient of 1.4. This apparent cooperativity at low ADP concentrations might serve to reduce the extent of catabolism of carbamoylphosphate under growth conditions yielding high energy charge. Experiments on the regulation of synthesis under various growth conditions showed a response to three regulatory signals: CKase was induced to high levels by anaerobiosis, induced to moderate levels by arginine, and repressed by ammonia. Thus, CKase expression is regulated in a manner that allows the enzyme to function as a provider of ammonia under aerobic conditions and of ATP under anaerobic conditions. ATP was an effective inhibitor of CKase activity; this inhibition provides the cell with an effective mechanism for avoiding a futile cycle resulting from the simultaneous operation of CKase and carbamoylphosphate synthetase when cells are grown in the presence of exogenous arginine. PMID- 6286600 TI - Cytosine methylation of the sequence GATC in a mycoplasma. AB - Mycoplasma virus L2 is subject to host-specific restriction and modification in Acholeplasma laidlawii strains JA1 and K2. We have examined the DNAs from both host cells and viruses propagated on these strains with respect to susceptibility to cleavage by restriction endonucleases and for DNA base modifications. We show that, in strain K2 and L2 virus grown on K2 cells, cytosine in the sequence GATC is methylated to 5-methylcytosine and, although strain K2 and L2 viruses grown on K2 contain N6-methyladenine in their DNA, adenine in the sequence GATC is not methylated. In contrast to K2, strain JA1 and L2 virus grown on JA1 cells contain no detectable methylated bases. It is not known which of the methylated bases in K2 is the basis for the K2 restriction-modification system operative on L2 virus. PMID- 6286601 TI - Novel phosphoenolpyruvate-dependent futile cycle in Streptococcus lactis: 2-deoxy D-glucose uncouples energy production from growth. AB - The addition of 2-deoxy-D-glucose to cultures of Streptococcus lactis 133 that were growing exponentially on sucrose or lactose reduced the growth rate by ca. 95%. Inhibition did not occur with glucose or mannose as the growth sugar. The reduction in growth rate was concomitant with rapid accumulation of the analog in phosphorylated form (2-deoxy-D-glucose 6-phosphate) via the phosphoenolpyruvate dependent mannose:phosphotransferase system. Within 5 min the intracellular 2 deoxy-D-glucose 6-phosphate concentration reached a steady-state level of greater than 100 mM. After maximum accumulation of the sugar phosphate, the rate of sucrose metabolism (glycolysis) decreased by only 30%, but the cells were depleted of fructose-1,6-diphosphate. The addition of glucose to 2-deoxy-D glucose 6-phosphate preloaded cells caused expulsion of 2-deoxy-D-glucose and a resumption of normal growth. S. lactis 133 contained an intracellular Mg2+ dependent, fluoride-sensitive phosphatase which hydrolyzed 2-deoxy-D-glucose 6 phosphate (and glucose 6-phosphate) to free sugar and inorganic phosphate. Because of continued dephosphorylation and efflux of the non-metabolizable analog, the maintenance of the intracellular 2-deoxy-D-glucose 6-phosphate pool during growth stasis was dependent upon continued glycolysis. This steady-state condition represented a dynamic equilibrium of: (i) phosphoenolpyruvate-dependent accumulation of 2-deoxy-D-glucose 6-phosphate, (ii) intracellular dephosphorylation, and (iii) efflux of free 2-deoxy-D-glucose. This sequence of events constitutes a futile cycle which promotes the dissipation of phosphoenolpyruvate. We conclude that 2-deoxy-D-glucose functions as an uncoupler by dissociating energy production from growth in S. lactis 133. PMID- 6286602 TI - Box-shaped halophilic bacteria. AB - Three morphologically similar strains of halophilic, box-shaped procaryotes have been isolated from brines collected in the Sinai, Baja California (Mexico), and southern California (United States). Although the isolates in their morphology resemble Walsby's square bacteria, which are a dominant morphological type in the Red Sea and Baja California brines, they are probably not identical to them. The cells show the general characteristics of extreme halophiles and archaebacteria. They contain pigments similar to bacteriorhodopsin which apparently mediate light driven ion translocation and photophosphorylation. PMID- 6286603 TI - A plasmid cloning vehicle for Haemophilus influenzae and Escherichia coli. AB - A new plasmid cloning vehicle (pDM2) was used to introduce a library of Haemophilus influenzae chromosomal fragments into H. influenzae. Transformants of the highly recombination-defective rec-1 mutant were more likely to contain exclusively recombinant plasmids after exposure to ligated DNA mixtures than was the wild type. pDM2 could replicate in Escherichia coli K-12. PMID- 6286606 TI - The proton translocating ATPase responsible for urinary acidification. AB - H+ secretion by the turtle urinary bladder is produced by a proton pump located in the luminal membrane. We show that a microsomal fraction of these cells contains an electrogenic proton translocating ATPase that is inhibited by dicyclohexylcarbodiimide but not oligomycin or vanadate. On a sucrose density gradient, this ATPase co-migrated with luminal membranes labeled with concanavalin A, but was separate from a lysosomal marker. This enzyme is therefore the H+ ATPase that causes urinary acidification. PMID- 6286605 TI - Inhibition of the phosphorylation of a 1000,000-dalton soluble protein in whole cells and cell-free extracts of PC12 pheochromocytoma cells following treatment with nerve growth factor. PMID- 6286604 TI - Relative map location of the rep and rho genes of Escherichia coli. AB - The rep gene of Escherichia coli was mapped between ilvC and rho by three-factor P1 transductional crosses and also by complementation with a set of lambda transducing phages that contain known amounts of bacterial DNA linked to ilvC. The physical distance between ilvC and rep and between rep and rho were calculated with an accuracy of +/- 0.4 kilobase to be 0 less than or equal to ilvC-rep less than or equal to 3.4 kilobases and 2.0 less than or equal to rep rho less than or equal to 6.0 kilobases. It was shown that rho-15 is Gro+ for phage ST-1. An ilv::Tn10 mutation was located in ilvY. PMID- 6286607 TI - Nerve growth factor induces specific enkephalin binding sites in a nerve cell line. AB - Specific enkephalin binding was found in a rat pheochromocytoma cell line, PC12 (subclone h), when cultured in the presence of nerve growth factor (NGF; 50 ng/ml). Specific binding of [3H][D-Ala2]Met-enkephalinamide was saturable with a KD=2.4 nM and Bmax = 866 fmol/mg of protein. The binding was displaced completely by morphine and naloxone at a concentration of 10(-4) and 10(-6) M, respectively. beta-Endorphin and dynorphin [1-13] also displaced the binding; KI = 9.6 and 81.6 nM, respectively. Enkephalin binding in PC12h cells cultured in the absence of NGF was very low. Binding was dramatically increased when cultured in the presence of NGF for 7 to 10 days. The enkephalin binding was induced by NGF in the cells cultured in a hormone-supplemented serum-free medium. In contrast, epidermal growth factor was of two orders lower potency, and insulin had no effect on the expression of enkephalin binding sites. Since the differentiation effects of NGF on PC12h cells are though to offer a model system to investigate the effect of NGH on neurons, the present observations may suggest a physiological role for NGF in the induction and maintenance of specific opiate receptors during development of the autonomic nervous system. PMID- 6286608 TI - The cytochrome c peroxidase activity of cytochrome oxidase. AB - I have found that mammalian cytochrome oxidase catalyzes the peroxidatic oxidation of ferrocytochrome c under strictly anaerobic conditions. An apparent Km value for ferrocytochrome c was 2 microM, and a second order rate constant, estimated as an extrapolated value, was 1.4 X 10(6) M-1 s-1 at pH 7.4 at 25 degrees C. These values were quite similar to the corresponding values of 6.4 microM and 1.9 X 10(6) M-1 s-1 determined for the intrinsic oxidase activity. The rate of the peroxidatic oxidation showed a hyperbolic dependence on the concentration of hydrogen peroxide, and the apparent Km value ws 0.18 mM. Cyanide and azide at 0.1 mM inhibited the peroxidase activity by 100 and 98%, respectively, whereas, under carbon monoxide at 750 mm Hg, 10% of the activity still remained. Under air, cytochrome oxidase acted simultaneously as oxidase and peroxidase. PMID- 6286609 TI - Quinine inhibition of Na+ and K+ transport provides evidence for two cation/H+ exchangers in rat liver mitochondria. AB - K+ efflux from respiring mitochondria via the K+/H+ exchanger is inhibited by quinine. In contrast, swelling in fresh mitochondria suspended in Na+ acetate, reflecting activity of the Na+/H+ exchanger, is unaffected by quinine. Swelling studies were also carried out in K+ and Na+ acetate in mitochondria loaded with tetraethylammonium and depleted of Mg2+ and K+. From the effects of pH and quinine on this preparation, we conclude that there are two distinct electroneutral pathways for monovalent cations. The Na+/H+ exchanger is highly specific for Na+ over K+ and is neither regulated by Mg2+ ions nor inhibited by quinine. The K+/H+ exchanger transports both Na+ and K+ and is regulated by Mg2+ ions and inhibited by quinine. PMID- 6286610 TI - Activation of Pseudomonas cytochrome oxidase by limited proteolysis with subtilisin. AB - Oxidized Pseudomonas cytochrome oxidase (ferrocytochrome c2: oxygen oxidoreductase; E.C.1.9.3.2) can be digested with subtilisin under controlled conditions that convert the original parent polypeptide chain (Mr on SDS gels approximately equal to 60,000) to a slightly smaller species (Mr on SDS gels approximately equal to 58,000). Under the conditions used (0.33% subtilisin, w/w, pH 7.4), the product formed from the oxidase was relatively stable to further digestion. Cytochrome oxidase activity was assayed at intervals during proteolysis by following the rate of oxidation of Pseudomonas ferrocytochrome c 551 by the enzyme in the presence of oxygen. The activity increased to a plateau that was more than two times the value for an untreated control. These observations suggest that clipping a small peptide from Pseudomonas cytochrome oxidase either facilitates the rate-limiting electron transfer between the intraprotein heme c and heme d1, enhances the interaction of the enzyme with ferrocytochrome c-551, or both. PMID- 6286611 TI - Determination of free and bound manganese(II) in hepatocytes from fed and fasted rats. PMID- 6286612 TI - Influence of the carboxyl terminus of luteinizing hormone-releasing hormone and bradykinin on hydrolysis by brain endo-oligopeptidases. AB - A homogeneous preparation of endo-oligopeptidase A from rabbit brain cleaves luteinizing hormone-releasing hormone (less than Glu-His-Trp-Ser-Tyr-Gly-Leu-Arg Pro-Gly-NH2) at the Tyr-Gly bond only after the removal of Gly-NH2 from the COOH terminal position of the molecule. The influence of the carboxyl terminus on hydrolysis by brain endo-oligopeptidases was studied using bradykinin as a model substrate. The substitution of the carboxyl group of bradykinin by the amide reduces by 2.5-fold the rate of Phe-Ser bond hydrolysis by endo-oligopeptidase A but has no effect on the rate of hydrolysis of the Pro-Phe bond by endo oligopeptidase B. On the other hand, the deletion of Phe-Arg from the COOH terminal portion of bradykinin makes the peptide resistant to hydrolysis by endo oligopeptidase A whereas it increases by 5-fold the rate of hydrolysis of the Pro Gly bond by endo-oligopeptidase B. PMID- 6286613 TI - Spectra of intermediates in oxidation and reduction of cytochrome c oxidase. AB - Two kinetic components with distinct difference spectra occur during reduction of cytochrome c oxidase by ruthenium hexamine. They are attributed to reduction of heme a (fast phase) and heme a3 (slow phase) (Scott, R. A., and Gray, H. B. (1980) J. Am. Chem. Soc. 102, 3219-3774). Two spectra seen during oxidation of cytochrome c oxidase by molecular oxygen have also been attributed to oxidation of hemes a3 and a (Greenwood, C., and Gibson, Q. H. (1967) J. Biol. Chem. 242, 1782-1787). We now report that spectra for the reductive and oxidative reactions obtained with the same preparations and the same apparatus under similar conditions are significantly different. The reactions appear to populate different reaction intermediates. Reconstitution into phospholipid vesicles does not affect these two spectra significantly. During turnover, the chief intermediates are those of the reductive pathway (Scott and Gray type intermediates). Reduction of heme a3 occurs approximately 70 times faster after turnover than the reduction of the resting enzyme. This is probably a dramatic "pulsing" effect (Wilson, M. T., Peterson, J., Antonini, E., Brunori, M., Colosimo, A., and Wyman, J. (1981) Proc. Natl. Acad. Sci. U.S.A. 7115-7118). PMID- 6286614 TI - Prolactin receptors in cultured rat mammary tumor cells. Energy-dependent uptake and degradation of hormone receptors. AB - After energy depletion by uncouplers of oxidative phosphorylation or inhibitors of electron transport, primary cultures of carcinogen-induced rat mammary tumors have a 2- to 20-fold increase in the number of cell surface prolactin receptors. When energy-depleted cells were treated with 0.15 M NaCl plus 50 mM glycine pH 3, for 1 min at 4 degrees C, 75% of the specific surface-bound 125I-labeled ovine prolactin was removed, but prolactin and its receptor were not destroyed. Using this technique, we found that receptor-bound prolactin can be internalized (becomes resistant to pH 3.0 treatment) and then degraded. The internalization of occupied receptors required energy, was completed 30-60 min before degradation, and was independent of protein synthesis. Hormone degradation (t1/2, 42 min) but not uptake was prevented by NH4Cl or lysosomotropic amines. In the presence of cycloheximide, receptors were lost (t1/2, 62 min) unless such loss was prevented by KCN. After unoccupied receptors were activated by energy depletion, surface receptors were lost when inhibitor was removed and glucose was added. Thus, both occupied and unoccupied prolactin receptors are constantly removed from the cell surface via an energy-dependent uptake mechanism. If the receptor levels are first increased by energy depletion (with or without bound ligand) or if protein synthesis is inhibited, there is a net loss of surface binding sites. Since the receptors reappeared with 15 h after cycloheximide removal, some of the receptors probably are recycled under normal steady state conditions. PMID- 6286615 TI - Structural studies of bovine heart cytochrome c1. AB - The complete primary structure of bovine heart cytochrome c1 was established by analyses of peptide fragments prepared by digestion using trypsin, staphylococcal protease, and chymotrypsin and by cyanogen bromide cleavage of cytochrome c1 and its derivatives. The total number of amino acid residues is 241, giving a molecular weight of 27,924 including the heme group. The NH2- and COOH-terminal residues are serine and lysine, respectively. One characteristic of the protein is that cytochrome c1 contains 43.6% hydrophobic residues and the polarity is estimated to be 41.1%. No clear homology was found between cytochrome c1 and other membranous proteins such as cytochrome b5 or the subunits of cytochrome oxidase for which sequences have been reported. Cytochrome c1 is predicted to have a high content of alpha-helix (46%). Partial sequence studies were also carried out on cytochrome c1 preparations obtained by different procedures and showed that there is no difference among the sequences of various preparations of cytochrome c1. The presence of a hydrophobic cluster near the COOH-terminal region indicates that the COOH-terminal region of cytochrome C1 associates with, or is buried in, the phospholipid bilayer of the mitochondrial membrane. PMID- 6286616 TI - Increased cyclic nucleotide phosphodiesterase activity in a mutant S49 lymphoma cell. Characterization and comparison with wild type enzyme activity. PMID- 6286617 TI - Structure and heme environment of ferrocytochrome c553 from 1H NMR studies. AB - Cytochrome c553 is a photosynthetic electron transport protein found in algae and cyanobacteria. We have purified cytochromes c553 from five cyanobacteria and studied the structures of the ferrocytochromes by 1H NMR spectroscopy at 360 and 470 MHz. Using standard NMR techniques and by comparing the amino acid sequences of four cytochromes c553 with their 1H NMR spectra, we have assigned in the spectrum of the Aphanizomenon flos-aquae protein 18 resonances to specific amino acid residues and 12 resonances to specific heme protons. Steady state and truncated driven nuclear Overhauser enhancement experiments indicate that a tyrosine and methionine are located near pyrrole ring IV of the heme and that a phenylalanine ring is near the heme alpha-mesoproton. The general folding of the cytochrome c553 protein backbone appears to resemble that of Pseudomonas aeruginosa cytochrome c551, but the chirality of the cytochrome c553 axial methine sulfur is R, the same as that of horse heart cytochrome c. PMID- 6286618 TI - Properties of proteoliposomes reconstituted with acetylcholine receptor from Torpedo californica. PMID- 6286619 TI - Reversible inhibition of the proton pump bacteriorhodopsin by modification of tyrosine 64. AB - Five mol of lysine per mol of bacteriorhodopsin were modified with methylacetimidate. This treatment did not inactivate bacteriorhodopsin but prevented all lysines from subsequent reaction with diazotized sulfanilic acid. This reaction predominantly modified tyrosine 64 and light-induced proton translocation was abolished. Reduction of the mono(p-azobenzene sulfonic acid) tyrosine 64 to the corresponding 3-amino derivative with sodium dithionite led to complete reactivation of the proton translocation activity of bacteriorhodopsin. The relative location of tyrosines 26 and 64 and the COOH terminus on the two surfaces of the purple membrane was determined by incorporation into phospholipid vesicles, subsequent modification, and proteolytic treatment. The results obtained support the models proposed by Engelmann et al. (Engelman, D. M., Henderson, R. McLauchlan, A. D., and Wallace, B. A. (1980) Proc. Natl. Acad. Sci. U. S. A. 77, 2023-2027) and by Ovchinnikov et al. (Ovchinnikov, Yu. A., Abdulaev, N. G., Feigina M. Yu., Kiselev A. V., and Lobanov, N. A. (1979) FEBS Lett. 100, 219-224). Tyrosine 64 is located on the extracellular side of the membrane, whereas tyrosine 26 and the COOH terminus are located on the cytoplasmic side. Because specific nitration of tyrosine 26 also leads to inactivation of bacteriorhodopsin (Lemke, H. D., and Oesterhelt, D. (1981) Eur. J. Biochem. 115, 595-604), the results obtained demonstrate that amino acid residues located on both surfaces of the purple membrane are involved in proton translocation. PMID- 6286620 TI - Purification by affinity chromatography of the glycine receptor of rat spinal cord. AB - The glycine receptor of rat spinal cord was solubilized with the nonionic detergent Triton X-100 and subsequently purified by affinity chromatography on aminostrychnine-agarose and wheat germ agglutinin-Sepharose. An overall purification of 1950-fold was achieved. Polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate and mercaptoethanol revealed three glycine receptor-associated polypeptides of Mr = 48,000, 58,000, and 93,000. [3H]Strychnine was incorporated irreversibly into the Mr = 48,000 polypeptide upon UV-illumination. The dissociation constant (KD) of [3H]strychnine binding to the purified glycine receptor was 9.3 +/- 0.6 nM. The glycine receptor agonists glycine, beta-alanine, and taurine inhibited the binding of [3H]strychnine to the purified receptor. Gel filtration and sedimentation in sucrose/H2O and sucrose/D2O gradients gave a Stokes radius of 7.7 nm, a partial specific volume of 0.780 +/- 0.005 ml/g and a sedimentation coefficient s20,w of 8.2 +/- 0.2 S for the purified glycine receptor. From these data, a molecular weight of 246,000 +/- 6,000 was calculated for the glycine receptor protein. PMID- 6286621 TI - The amiloride-sensitive Na+/H+ exchange system in skeletal muscle cells in culture. AB - Chick skeletal muscle cells in culture have an amiloride-sensitive Na+ transporting system that has the following properties. Na+ uptake is dependent on the extracellular Na+ concentration. The Km value for Na+ is 25 mM and remains constant between pH 7.5 and 8.5. The maximal rate of Na+ transport is higher at alkaline pH. An ionizable group with a pK of 7.6 is essential for the system to be functional. The activity of the amiloride-sensitive Na+ uptake system is controlled by internal Na+ and H+ concentrations. Amiloride inhibition of Na+ uptake is competitively antagonized by increasing Na+ concentration. The dissociation constant for amiloride is 5 microM in Na+-free conditions and is constant between pH 7.5 and 8.5. The Km value for Na+ found from competition experiments is 13 mM. The amiloride-sensitive Na+ influx occurs in parallel with an amiloride-sensitive H+ efflux. This H+ efflux is stimulated by increasing external Na+ concentrations, the Km for Na+ being 15 mM. It is inhibited by amiloride with the same concentration dependence as Na+ influx. PMID- 6286622 TI - Solubilization of active prolactin receptors by a nondenaturing zwitterionic detergent. PMID- 6286623 TI - Epinephrine regulation of phosphofructokinase in perfused rat heart. A calcium ion-dependent mechanism mediated via alpha-receptors. PMID- 6286624 TI - Structure of catabolite gene activator protein at 2.9-A resolution. Incorporation of amino acid sequence and interactions with cyclic AMP. AB - The amino acid sequence of the Escherichia coli catabolite gene activator protein has been fit into a 2.9-A resolution electron density map. Each subunit of the dimer consists of two structurally distinct domains. The larger NH2-terminal domain is seen to bind cyclic AMP and forms all of the contacts between the subunits. The cyclic AMP is completely buried between the interior of the "beta roll" structure of the large domain and a long alpha helix; it makes important hydrogen-bonding interactions with residues from both subunits. The guanidinium group of a buried Arg makes an internal salt link with the phosphate of cyclic AMP. The 6-amino group of adenine interacts simultaneously with both subunits. This interaction with both subunits and the fact that cyclic GMP and cyclic IMP do not activate catabolite gene activator protein suggest that the binding of cyclic AMP may alter the relative orientation of the two subunits, which in turn would change the structure of a DNA binding site that is presumed to span the two smaller domains. The distribution and nature of side chains in the small domain do not rule out the possibility that catabolite gene activator protein binds to left-handed B-DNA. PMID- 6286625 TI - Modulation of rat parotid cell alpha-adrenergic responsiveness at a step subsequent to receptor activation. AB - Parotid gland acinar cells, prepared from 12- and 24-month-old rats, show decreased physiological responsiveness to alpha-adrenergic stimulation in vitro compared to cells from 3-month-old rats. K+ efflux, an index of water and electrolyte secretion, was approximately 35% lower with 12- and 24-month-old parotid cells. No loss of alpha-adrenergic receptors, their binding affinity for specific alpha-adrenergic ligands, or their relative subtype distribution, accompanied the diminished exocrine function. Conversely, a significant reduction in alpha-adrenergic-mediated phospholipid turnover in, and 45Ca2+ efflux from, parotid cells of older rats was observed. These changes in phospholipid metabolism and Ca2+ flux were parallel to changes seen in K+ efflux as judged by dose-response studies. When the alpha-adrenergic receptor was by-passed by using the Ca2+-ionophore A-23187 to elicit K+ efflux, young and old parotid cells were equally responsive. In aggregate the findings suggest that parotid gland cells from older rats display an altered alpha-adrenergic signal transduction mechanism at a site between the receptor and phospholipid turnover/Ca2+ mobilization. PMID- 6286626 TI - Mossbauer and EPR studies on nitrite reductase from Thiobacillus denitrificans. PMID- 6286627 TI - Cyclic AMP-dependent phosphorylation of terminal deoxynucleotidyl transferase. AB - Incubation of human lymphoblastoid cell extracts in the presence of cAMP and ATP produces changes in the chromatographic pattern of terminal transferase activity separated on phosphocellulose columns. Incubation of high molecular weight and low molecular weight preparations of calf thymus terminal transferase with the catalytic subunit of beef cardiac muscle cAMP-dependent protein kinase and [gamma 32P]ATP result in phosphorylation of the 58,000-dalton form of the enzyme and no other lower molecular weight terminal peptides. These results, taken with our earlier results on tissue proteolysis of terminal transferase to lower molecular weight, active forms (Chang, L. M. S., Plevani, P., and Bollum, F. J. (1982) J. Biol. Chem. 257, 5700-5706), resolve the heterogeneity observed with various preparations of the enzyme. PMID- 6286628 TI - Role of the divalent metal cation in the pyruvate oxidase reaction. AB - Purified pyruvate oxidase requires a divalent metal cation for enzymatic activity. The function of the divalent metal cation was studied for unactivated, dodecyl sulfate-activated, and phosphatidylglycerol-activated oxidase. Assays performed in the presence of Mg2+, CA2+, Zn2+, Mn2+, Ba2+, Ni2+, Co2+, Cu2+, and Cr3+ in each of four different buffers, phosphate, 1,4-piperazinediethanesulfonic acid, imidazole, and citrate, indicate that any of these metal cations will fulfill the pyruvate oxidase requirement. Extensive steady state kinetics data were obtained with both Mg2+ and Mn2+. All the data are consistent with the proposition that the only role of the metal is to bind to the cofactor thiamin pyrophosphate (TPP) and that it is the Me2+-TPP complex which is the true cofactor. Values of the Mg2+ and Mn2+ dissociation constants with TPP were determined by EPR spectroscopy and these data were used to calculate the Michaelis constant for the Me2+-TPP complexes. The results show that the Michaelis constants for the Me2+-TPP complexes are independent of the metal cation in the complex. Fluorescence quenching experiments show that the Michaelis constant is equal to the dissociation constant of the Mn2+-TPP complex with the enzyme. It was also shown that Mn2+ will only bind to the enzyme in the presence of TPP and that one Mn2+ binds per subunit. Steady state kinetics experiments with Mn2+ were more complicated than those obtained with Mg2+ because of the formation of an abortive Mn2+-pyruvate complex. Both EPR and steady state kinetics data indicated complex formation with a dissociation constant of about 70 mM. PMID- 6286629 TI - 23S,25-dihydroxyvitamin D3 as a circulating metabolite of vitamin D3. Its role in 25-hydroxyvitamin D3-26,23-lactone biosynthesis. AB - 23,25-Dihydroxyvitamin D3 was isolated from vitamin D-toxic pig plasma by sequential chromatography through two gravity columns and three high performance liquid chromatography systems. Two of the high performance liquid chromatography systems separated the R and S diastereomers of 23,25-dihydroxyvitamin D3 and demonstrated that the metabolite has the 23S configuration. Ultraviolet absorbance and mass spectroscopy of the pure metabolite and mass spectroscopy of its trisilylated derivative confirmed the structural assignment. 23S,25 Dihydroxyvitamin D3, 23R,25-dihydroxyvitamin D3, 25S,26-dihydroxyvitamin D3, and 25R,26-dihydroxyvitamin D3 were assessed for their ability to produce 25 hydroxyvitamin D3-26,23-lactone in vitamin D2-toxic rats. On 23S,25 dihydroxyvitamin D3, of the naturally occurring compounds, was able to increase the plasma lactone concentration. This metabolite was a more efficient precursor than 25-hydroxyvitamin D3, suggesting that 23S-hydroxylation is a rate-limiting step in 25-hydroxyvitamin D3-26,23-lactone formation. 23S,25-Dihydroxyvitamin D3 was not detected in 25-hydroxyvitamin D3-dosed rats, indicating that the former is rapidly metabolized. Nephrectomized rats had a diminished but significant ability to synthesize 25-hydroxyvitamin D3-26,23-lactone from 25-hydroxyvitamin D3. Nephrectomy did not affect synthesis of 25-hydroxyvitamin D3-26,23-lactone from 23S,25-dihydroxyvitamin D3. These results demonstrate that vitamin D3 23S hydroxylase(s) are also located extrarenally and that extrarenal tissues are quantitatively important to 25-hydroxyvitamin D3-26,23-lactone synthesis. PMID- 6286630 TI - Photoaffinity labeling of thyroid hormone nuclear receptors in intact cells. PMID- 6286631 TI - Cytochalasin B enhances hormone and cholera toxin-stimulated cyclic AMP accumulation in S49 lymphoma cells. PMID- 6286632 TI - Complete analysis of cellular nucleotides by two-dimensional thin layer chromatography. AB - We describe methods for the complete analysis of cellular nucleotides from as few as 10(6) 32Pi-labeled cells in a simple 2-day experiment. Nucleotides are extracted with acid, neutralized, and resolved by two-dimensional thin layer chromatography on polyethyleneimine cellulose. In the first dimension the nucleotides are separated based on the negative charge of their phosphate groups (i.e. cyclic, mono-, di, and triphosphates) and in the second dimension on their content of nucleobases (i.e. Ura, Cyt, Thy, Gua, and Ade). Because the separation is logical, one can predict the chromatographic migration of most nucleotides. By running standards we have determined the chromatographic location of over 90 biologically important nucleotides, nucleotide derivatives, and modified nucleotides from tRNA. We also developed a set of enzymatic and chemical methods to be used in conjunction with the chromatographic separations for verifying the identity of nucleotides and characterizing novel nucleotides. In this paper we use these methods to analyze and inventory the nucleotide content of Salmonella typhimurium in balanced log phase growth. Other potential uses of the method are also described. PMID- 6286634 TI - Differential accumulation of two apo-iso-cytochromes c in processing mutants of yeast. PMID- 6286633 TI - Biochemical and genetic studies of the apoprotein E secreted by mouse macrophages and human monocytes. PMID- 6286635 TI - Proteolytic cleavage of epidermal growth factor receptor. A Ca2+-dependent, sulfhydryl-sensitive proteolytic system in A431 cells. AB - The Mr = 160,000 epidermal growth factor (EGF) receptor in A431 cells is partially cleaved during membrane isolation to a Mr = 145,000 polypeptide containing both EGF binding and phosphate acceptor sites. We show that the proteolytic degradation of the EGF receptor depends upon the presence of Ca2+ in the medium used to scrape the cells from the substratum. Only the high molecular weight form of the receptor is detected in membranes prepared in the absence of Ca2+. Ca2+-dependent proteolysis occurs rapidly (t1/2 approximately 5 min) following cell scraping. Proteolysis results in a decrease in EGF-dependent phosphorylation of the receptor while retaining EGF binding capacity. In addition, membranes containing the uncleaved form of the receptor reveal a substantial increase in EGF-dependent phosphorylation of proteins with Mr approximately 80, 89, and 185 X 10(3). In the presence of Ca2+, addition of iodoacetic acid to the scraping medium strongly inhibits receptor fragmentation, whereas other inhibitors (phenylmethylsulfonyl fluoride, leupeptin, and pepstatin) have no effect. The results implicate a role for a Ca2+-dependent, SH sensitive protease in EGF receptor degradation. Prevention of proteolysis yields membrane preparations with highly active EGF-dependent kinase system. PMID- 6286636 TI - Interactions between the receptors for insulin and the insulin-like growth factors on adipocytes. PMID- 6286637 TI - Isolation, characterization, and comparison of the solubilized particulate and soluble folate binding proteins from human milk. PMID- 6286638 TI - Characterization of eight forms of corticotropin-like intermediary lobe peptide from the rat intermediary pituitary. AB - The isolation and characterization of eight forms of corticotropin-like intermediary lobe peptide (CLIP, adrenocorticotropin18-39) from the intermediary lobe of the rat pituitary has been accomplished by using reversed phase high performance liquid chromatography. The eight forms are the result of all combinations of the presence or absence of three post-translational modifications. These are glycosylation, phosphorylation, and removal of the carboxyl-terminal amino acid. The sites of phosphorylation and glycosylation are at serine 31 and asparagine 29, respectively. The eight forms (in order of elution from the reversed high performance liquid chromatography column) are glycosylated, phosphorylated CLIP18-38; glycosylated, nonphosphorylated CLIP18 38; nonglycosylated, phosphorylated CLIP18-38; nonglycosylated, nonphosphorylated CLIP18-38; glycosylated, phosphorylated CLIP18-39; glycosylated, nonphosphorylated CLIP18-39; nonglycosylated, phosphorylated CLIP18-39; and nonglycosylated, nonphosphorylated CLIP18-39. PMID- 6286639 TI - A thermodynamic study of ordered complexes of cytochrome c fragments. Kinetic and equilibrium measurements and comparison with the folding of the intact protein. PMID- 6286640 TI - Effect of tunicamycin on the synthesis, processing, and secretion of pro opiomelanocortin peptides in mouse pituitary cells. AB - Pro-opiomelanocortin (POMC) is glycosylated and proteolytically cleaved to produce a number of smaller peptide hormones including adrenocorticotropic hormone (ACTH) and endorphin in mammalian pituitary and the mouse pituitary cell line AtT-20/D16v. When glycosylation of POMC is inhibited in AtT-20 cells with the drug tunicamycin, a 26,000-dalton protein appears in place of the glycosylated 29,000- and 32,000-dalton forms of POMC. The 26,000-dalton form found in tunicamycin-treated cells has the same [35S]methionine tryptic peptides as 29,000- and 32,000-dalton POMC, indicating that the decrease in apparent mass is most likely due to loss of carbohydrate and not to changes in the peptide backbone. The 4,500-dalton form of alpha(1-39)ACTH and the 3,000- and 11,000 dalton forms of endorphin are all present in tunicamycin-treated cells. The glycosylated form of alpha(1-39)ACTH, however, is missing and the glycosylated ACTH intermediates are replaced by unglycosylated ACTH intermediates. Pulse-chase studies demonstrate that the 26,000-dalton unglycosylated POMC is the precursor of the smaller ACTH and endorphin molecules in tunicamycin-treated cells. Furthermore, all of the forms of ACTH and endorphin found in tunicamycin-treated cells are secreted. Thus, it appears that glycosylation is not an essential step for correct cleavage or secretion of POMC or its products. PMID- 6286641 TI - Left-handed DNA. Cloning, characterization, and instability of inserts containing different lengths of (dC-dG) in Escherichia coli. AB - Recombinant pBR322 derivatives were constructed containing tracts of (dC-dG) sequences which are 58, 32, 26, and 10 base pairs (bp) in length in conjunction with a 95-bp fragment containing the Escherichia coli lac operator-promoter. The biological properties of these plasmids were unusual since deletions in the (dC dG) regions, but not in the pBR322 nor the lac segments, were frequently observed; segments of (dC-dG) longer than approximately 50 bp were not stable but suffered deletions. Segments of approximately 30 bp or shorter were stable in most cases. The (dC-dG) tracts seemed to enhance recA-mediated recombination when they were of suitable length and were cloned into certain sites in the recombinants. Also the (dC-dG)-containing plasmids were less supercoiled (by 6-12 turns) than expected, relative to control plasmids, after isolation from E. coli hosts. These recombinant plasmids were used in the following paper to evaluate the properties of segments containing the unorthodox left-handed conformations. PMID- 6286642 TI - Energetic and structural inter-relationship between DNA supercoiling and the right- to left-handed Z helix transitions in recombinant plasmids. AB - We have evaluated the B to Z conformational transitions in supercoiled recombinant plasmids containing different lengths of (dC-dG) described in the preceding paper. The sodium chloride-induced right- to left-handed transition in a small segment of the plasmids caused a relaxation of (-) supercoils which was monitored by electrophoretic mobility changes of individual topoisomers on agarose gels containing NaCl at concentrations up to 5.0 M. The number of supercoils relaxed was proportional to the length of the (dC-dG) segment in the plasmid in good agreement with theoretical values. A short B/Z junction region (less than 5 base pairs) was inferred. The stability of the Z conformation in (dC dG) segments of the plasmids had a strong length dependency; shorter lengths were less stable. Ten base pairs of (dC-dG) was insufficient to allow a Z conformation under the conditions studied. Supercoiling imparts a substantial favorable free energy to the Z conformation, reducing the NaCl concentration necessary to cause the transition. The relationship of supercoiling with the NaCl concentration necessary to cause a B leads to Z transition suggests that supercoiling alone is sufficient to stabilize the Z conformation at physiological salt concentrations. These results support the notion that left-handed DNA has an important biological role. PMID- 6286643 TI - Temporal aspects of the N- and O-glycosylation of human chorionic gonadotropin. AB - The glycoprotein hormone, human chorionic gonadotropin (hCG), contains both N- and O-linked oligosaccharide chains linked to its beta-subunit. Using the human choriocarcinoma cell line, BeWo, we have examined the temporal relationship between N- and O-glycosylation of hCG and the subsequent processing of both types of oligosaccharide chains. The results indicate that, as observed in related cell lines, mature, completely glycosylated forms of the subunits of hCG cannot be detected intracellularly in BeWo cells during pulse-chase experiments with [35S]methionine. To more directly study the temporal relationship between N- and O-glycosylation of hCG in BeWo cells, 14C-amino acids and [3H]glucosamine (which also serves as a precursor to N-acetylgalactosamine) were used to label hCG. The results of these studies are consistent with a model for the N- and O glycosylation of hCG in which 1) N-glycosylation of hCG occurs co-translationally or very shortly after translation, and 2) the addition of O-linked GalNAc residues to the polypeptide and the addition of peripheral GlcNAc residues to the N-linked oligosaccharide chains occur just prior to secretion, presumably in the Golgi complex. PMID- 6286644 TI - The interaction of arylazido ubiquinone derivative with mitochondrial ubiquinol cytochrome c reductase. PMID- 6286645 TI - Characterization of the DNA polymerases induced by a group of herpes simplex virus type I variants selected for growth in the presence of phosphonoformic acid. AB - Five independently derived variants of a herpes simplex virus type I (HSV-1) strain were plaque purified from a virus population passaged in 1 mM phosphonoformic acid (PFA). The DNA polymerase induced by the parent and PFA resistant viruses were purified and characterized. No differences were observed among the enzymes with respect to their chromatographic properties, specific activities, or polypeptides resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The variant enzymes exhibited levels of PFA resistance which ranged from 15- to 25-fold. Resistance to PFA was always associated with a similar degree of resistance to its congener phosphonoacetic acid, but cross resistance to beta-phenylphosphonoacetic acid was only seen with two of the five variant enzymes. PFA and pyrophosphate were mutually competitive in PPi exchange reactions, but in DNA synthetic reactions the levels of resistance to PFA and PPi were not equal. The apparent affinities of the enzymes for Mg2+ did parallel their affinities for PFA. Km values of dNTPs were about 2-fold higher than the parent virus enzyme for all of the variant enzymes except one which was 4-fold higher. The processivity of polymerization was apparently unaffected by the enzyme changes related to PFA resistance although one variant enzyme had a lower value. Resistance among the variant enzymes to the triphosphates of 9-(2 hydroxyethoxymethyl)guanine and 2',3'-dideoxyguanosine was directly related to the level of resistance to PFA. The data presented here indicated that (i) PFA resistance may result from several types of active site alterations, since the PFA-resistant enzymes were of three kinetically distinct types. Also, additional enzyme alterations, probably unrelated to PFA resistance, were detected in one enzyme. (ii) PFA and PPi possess some different binding determinants within the active center of herpes simplex virus type I DNA polymerase. (iii) PFA and the triphosphates of 9-(2-hydroxyethoxymethyl)guanine and 2',3'-dideoxyguanosine may have a common ultimate inhibitory mechanism. PMID- 6286647 TI - Isolation, characterization, and amino acid sequence of a ubiquitin-like protein from insect eggs. AB - A protein with a primary structure identical to that of human and bovine ubiquitin has been purified from insect eggs. The isolation, secondary structure, and amino acid sequence of this ubiquitin-like protein are reported. The sequence was determined by automatic Edman degradation of the intact molecule as well as by the manual sequence analysis of the enzymatic cleavage products. The polypeptide has 74 amino acid residues and internal homology regions. Interactions of the protein with peptides results in protective effects against proteolysis. This paper reports for the first time the presence of the ubiquitin molecule in invertebrates. PMID- 6286646 TI - Selective antiviral agents. The metabolism of 5-propyl-2'-deoxyuridine and effects on DNA synthesis in herpes simplex virus type 1 infections. AB - The metabolism and disposition of 5-propyl-2'-deoxyuridine (Pr-dUrd) in herpes simplex virus type 1 infections were investigated in cell culture using [14C]Pr dUrd, [32P]orthophosphate, and several methods including high pressure liquid chromatography and isopycnic centrifugation. Results in infected cells indicate Pr-dUrd 1) is taken up and phosphorylated to mono-, di-, and triphosphates; 2) is incorporated into DNA; 3) preferentially inhibits synthesis of viral DNA; 4) blocks re-initiation of viral DNA synthesis even after removal of the nucleoside from the culture; and 5) exerts these effects early in the course of infection (before 6 h postinfection). Pr-dUrd was not phosphorylated in uninfected cells, and had little or no effect on apparent cellular DNA synthesis in infected or uninfected cells. Present evidence suggests one possible antiviral event could be the lethal effect of Pr-dUrd after incorporation into viral DNA by alteration of DNA template-directed functions such as replication. PMID- 6286648 TI - Electron spin echo studies of the copper complexes of conalbumin. AB - Electron spin echo envelope spectroscopy was used to probe the two metal binding sites of Cu(II)-conalbumin. The echo envelope spectrum of Cu(II)-conalbumin oxalate, with metal ion at either one or both of the binding sites, contains lines arising from the interaction of the electron spin of Cu(II) with bound imidazole, demonstrating histidine ligation to the metal ion. The 13C superhyperfine interaction of bound [13C]oxalate, obtained from the ratio of the electron spin echo envelopes of Cu(II)-conalbumin-[13C]oxalate to that of Cu(II) conalbumin-[12C]oxalate, is about twice the free precession frequency and indicates a contact interaction between 13C and Cu(II). This study indicates that oxalate is directly coordinated to the metal ion. Over the pH range 7.0 to 10.0, where Cu(II)-conalbumin binds carbonate as an associated anion, the echo envelope spectrum indicates that at least one imidazole ligand is coordinated to Cu(II). Below pH 6.0 and above pH 11.0, imidazole coordination is not observed. PMID- 6286649 TI - Regulation of HeLa cell transferrin receptors. AB - HeLa cells were found to have a single class of non-interacting receptors specific for transferrin. Both apotransferrin and diferric transferrin competed equally with 125I-diferric transferrin for receptor binding. Transferrin binding was temperature-dependent and reversible. Binding of transferrin to cells exhibited a KD of 27 nM with a maximum binding capacity of 1.8-3.7 x 10(6) molecules/cell. Cells grown in the presence of diferric transferrin or in the presence of ferric ammonium citrate exhibited a concentration- and time-dependent decrease in 125I-diferric transferrin binding. The decrease in binding activity reflected a reduction in receptor number rather than an alteration in ligand receptor affinity. Growth of cells in saturating concentrations of apotransferrin did not cause a decrease in receptor number. When iron-treated cells were removed to media free of ferric ammonium citrate, the receptor number returned to control values by 40 h. When receptors were removed with trypsin, cells grown and maintained in ferric ammonium citrate-supplemented media demonstrated a rate of receptor reappearance 47% that of control cells grown in ferric ammonium citrate free media. Cells grown in media supplemented with diferric transferrin or ferric ammonium citrate exhibited an increase in cytosolic iron content. The transferrin receptor number returned to normal after cells were removed to unsupplemented media, despite persistent elevation of cytosolic iron content. Increased iron content did not appear to be the sole factor determining receptor number. PMID- 6286650 TI - The mechanism of ubiquitin activating enzyme. A kinetic and equilibrium analysis. PMID- 6286651 TI - Hormonal regulation of (Na+,K+)-ATPase biosynthesis in the toad bladder. Effect of aldosterone and 3,5,3'-triiodo-L-thyronine. AB - Aldosterone stimulates transepithelial Na+ transport in the toad bladder, and thyroid hormone antagonizes this mineralocorticoid action. In the present study, we assessed the influence of these two hormones on the biosynthesis of (Na+,K+)ATPase, the major driving force of Na+ transport. Rates of enzyme synthesis were estimated by immunoprecipitation with monospecific alpha (96,000 daltons) and beta (60,000 daltons) subunit antibodies. After a 30-min pulse of intact tissue with [35S]methionine, the anti-alpha-serum recognized the 96,000 dalton alpha subunit and the anti-beta-serum, a 42,000-dalton protein, in total cell extracts. The biosynthesis rates of both these proteins were increased 2.8- and 2.4-fold respectively, over controls by 80 nM aldosterone after 18 h of hormone treatment. The hormonal effect was not apparent up to 3 h of incubation and was dose dependent between 0.2 and 20 nM aldosterone. The hormonal induction was antagonized by spironolactone (500-fold excess) but not by amiloride. The action of aldosterone thus seems to be a receptor-mediated process and a primary event independent of the Na+ permeability of the apical membrane. Thyroid hormone, on the other hand, had no effect on either basal or aldosterone stimulated synthesis rates of both enzyme proteins. The results demonstrate a direct effect of aldosterone on gene expression of the (Na+,K+)-ATPase. Ultimately, this phenomenon could be linked to the late mineralocorticoid action of this hormone. On the other hand, thyroid hormone, in contrast to the situation in mammals, does not stimulate de novo enzyme synthesis in amphibia. Neither can the antimineralocorticoid action of thyroid hormone in the toad bladder be explained by an inhibition of the (Na+,K+)-ATPase synthesis. PMID- 6286652 TI - Biogenesis of mitochondrial ubiquinol:cytochrome c reductase (cytochrome bc1 complex). Precursor proteins and their transfer into mitochondria. AB - The precursor proteins to the subunits of ubiquinol:cytochrome c reductase (cytochrome bc1 complex) of Neurospora crassa were synthesized in a reticulocyte lysate. These precursors were immunoprecipitated with antibodies prepared against the individual subunits and compared to the mature subunits immunoprecipitated or isolated from mitochondria. Most subunits were synthesized as precursors with larger apparent molecular weights (subunits I, 51,500 versus 50,000; subunit II, 47,500 versus 45,000; subunit IV (cytochrome c1), 38,000 versus 31,000; subunit V (Fe-S protein), 28,000 versus 25,000; subunit VII, 12,000 versus 11,500; subunit VIII, 11,600 versus 11,200). Subunit VI (14,000) was synthesized with the same apparent molecular weight. The post-translational transfer of subunits I, IV, V, and VII was studied in an in vitro system employing reticulocyte lysate and isolated mitochondria. The transfer and proteolytic processing of these precursors was found to be dependent on the mitochondrial membrane potential. In the transfer of cytochrome c1, the proteolytic processing appears to take place in two separate steps via an intermediate both in vivo and in vitro. In vivo, the intermediate form accumulated when cells were kept at 8 degrees C and was chased into mature cytochrome c1 at 25 degrees C. Both processing steps were energy dependent. PMID- 6286653 TI - Type II regulatory subunit of cAMP-dependent protein kinase. Phosphorylation by casein kinase II at a site that is also phosphorylated in vivo. AB - A study of the phosphorylated sites on the regulatory subunit of type II cyclic AMP-dependent protein kinase is described. The site of autophosphorylation, previously reported (Takio, K., Walsh, K. A., Neurath, H., Smith, S. B., Krebs, E. G., and Titani, K. (1980) FEBS Lett. 114, 83-88), is found to contain the only residue phosphorylated by the catalytic subunit. Endogenous protein-bound phosphate, present at a second site, is observed in the NH2-terminal region of purified type II regulatory subunit. A serine residue in this same domain of type II regulatory subunit is shown to be phosphorylated in vitro by casein kinase II. The stoichiometry of phosphate incorporation by casein kinase II indicates a single site of phosphorylation that is equivalent to the site containing the endogenous phosphate. The apparent Km and Vmax of casein kinase II for the phosphorylation of purified type II regulatory subunit containing 0.55 molar equivalents of endogenous phosphate are 36 microM and 3.5 mumol/min/mg, respectively. Phosphatase treatment of type II regulatory subunit decreased the determined Km to 13 microM but does not change the Vmax consistent with the suggestion that casein kinase II is phosphorylating a site partially occupied by endogenous phosphate. Comparison of peptide maps of type II regulatory subunit phosphorylated in vitro by casein kinase II with in vivo modified type II regulatory subunit indicates that the site modified in vitro is also phosphorylated in vivo. PMID- 6286655 TI - Characterization of reversible, physical binding of benzo[a]pyrene derivatives to DNA. AB - The carcinogen, 7r,8t-dihydroxy-9t,10t-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene, and a number of its noncarcinogenic derivatives form a reversible physical complex with DNA in vitro. The absorbance spectrum of the hydrocarbon in the complex shifts approximately 10 nm to the red. Fluorescence from the physically bound hydrocarbon is significantly quenched, and binding levels are decreased by the addition of 1) spermine, 2) MgCl2, and 3) NaCl, with their relative effectiveness in reducing complex formation following the same order. Equilibrium data and the change in superhelicity of simian virus 40 DNA as a result of physical binding were used to calculate an unwinding angle for each of the benzo[a]pyrene derivatives of 13 degrees, while under the same conditions ethidium bromide unwound the duplex approximately 30 degrees. The weight of this evidence indicates that the carcinogen and its noncarcinogenic derivatives reversibly bind to DNA by intercalation of their planar aromatic system into the stacked base pairs of the duplex. The results of this and previous studies suggest that covalent binding of the carcinogenic derivative of benzo[a]pyrene to DNA is preceded by a physical intercalation step. PMID- 6286654 TI - Evidence that human platelet alpha-adrenergic receptors coupled to inhibition of adenylate cyclase are not associated with the subunit of adenylate cyclase ADP ribosylated by cholera toxin. AB - Exposure of the alpha-adrenergic receptor of the human platelet to agonist prior to solubilization stabilizes a receptor complex of the alpha-adrenergic receptor with the GTP-binding protein(s) which modulates receptor affinity for agonists (Smith, S. K., and Limbird, L. E. (1981) Proc. Natl. Acad. Sci. U. S. A. 78, 4026 4030). The soluble alpha-adrenergic receptor is characterized by retention of sensitivity to GTP and a faster rate of sedimentation in sucrose gradients than antagonist-occupied or unoccupied receptors. The present studies were undertaken to determine whether the alpha-adrenergic receptor, which is coupled to inhibition of adenylate cyclase, contains the same GTP-binding protein that is involved in activation of adenylate cyclase. The GTP-binding protein that is coupled to activation of adenylate cyclase was labeled with [32P]ADP-ribose using cholera toxin. Incorporation of [32]ADP-ribose into a Mr = 42,000 peptide in human platelet membranes was paralleled by an enhancement of GTP-sensitive catalytic activity in the membranes. However, cholera toxin treatment did not modify alpha-receptor-mediated inhibition of adenylate cyclase or interaction of the alpha-receptor with agonist agents. Moreover, sucrose gradient centrifugation revealed that the [32P]ADP-ribosylated Mr = 42,000 subunit of the stimulatory GTP binding protein did not appear to associate with the agonist-alpha-receptor complex. These data suggest that the GTP-binding protein that mediates GTP activation of adenylate cyclase in the human platelet membrane is distinct from the GTP-binding protein that modulates alpha-adrenergic receptor affinity for agonist agents and which associates with the receptor in the presence of agonists. PMID- 6286656 TI - Acetylation of alpha-melanotropin and beta-endorphin in the rat intermediate pituitary. Subcellular localization. AB - The subcellular site of the further processing (NH2-terminal acetylation and COOH terminal proteolysis) of beta-endorphin-sized molecules in the rat intermediate pituitary has been studied. Rat intermediate pituitary primary cultures that had been incubated in radioactively labeled amino acids were homogenized and the secretory granule fraction was separated from the rough endoplasmic reticulum/Golgi apparatus fraction. The labeled beta-endorphin-sized molecules in each subcellular fraction were analyzed by immunoprecipitation, gel filtration chromatography, and ion exchange chromatography. A large percentage of the labeled beta-endorphin-sized molecules became NH2 terminally acetylated after becoming associated with the secretory granule fraction; most of the further COOH terminal proteolytic processing of alpha-N-acetyl-beta-endorphin(1-31) to form alpha-N-acetyl-beta-endorphin(1-27) and alpha-N-acetyl-beta-endorphin(1-26) also occurred when the labeled beta-endorphin-sized molecules were associated with the secretory granule fraction. The acetylation of alpha-melanocyte-stimulating hormone (alpha MSH)-sized molecules was also investigated in rat intermediate pituitary primary cell cultures by immunoprecipitation, gel filtration chromatography, and reverse-phase high pressure liquid chromatography. Pulse chase labeling experiments showed that newly synthesized molecules co-migrating with adrenocorticotropic hormone ((ACTH)(1-13)NH2) were converted first to molecules similar to alpha MSH (alpha-N-acetyl-ACTH(1-13)NH2) and then to molecules similar to alpha-N,O-diacetyl-alpha MSH. These results demonstrate that the enzyme activity(s) responsible for the NH2-terminal acetylation of beta endorphin alpha MSH-sized molecules is located in the secretory granules of the rat intermediate pituitary. PMID- 6286657 TI - Characterization of the peptide acetyltransferase activity in bovine and rat intermediate pituitaries responsible for the acetylation of beta-endorphin and alpha-melanotropin. PMID- 6286658 TI - Cerulenin blocks fatty acid acylation of glycoproteins and inhibits vesicular stomatitis and Sindbis virus particle formation. AB - Cerulenin, an antibiotic that inhibits de novo fatty acid and cholesterol biosynthesis, effectively inhibited the formation and release of virus particles from chicken embryo fibroblasts infected with Sindbis or vesicular stomatitis virus (VSV). When added for 1 h at 3 h postinfection, the antibiotic blocked VSV particle production by 80 to 90% and inhibited incorporation of [3H]palmitic acid into the VSV glycoprotein by an equivalent amount. The effect of this antibiotic on virus protein and RNA biosynthesis was significantly less than that on fatty acid acylation. Nonacylated virus glycoproteins accumulated inside and on the surface of cerulenin-treated cells. These data indicate that fatty acid acylation is not essential for intracellular transport of these membrane proteins, but it may have an important role in the interaction of glycoproteins with membranes during virus assembly and budding. PMID- 6286659 TI - Binding of phosphorylated oligosaccharides to immobilized phosphomannosyl receptors. AB - We have purified phosphomannosyl-enzyme receptors from bovine liver on an affinity column composed of glycoproteins isolated from Dictyostelium discoideum secretions. Binding of human fibroblast beta-hexosaminidase B to receptors reconstituted into phosphatidylcholine liposomes was 1) specifically inhibited by mannose 6-phosphate, but not mannose 1-phosphate or glucose 6-phosphate, and 2) had properties similar to the previously reported binding of enzyme to receptors on cell surfaces and isolated membranes. In order to determine the structural features of the phosphomannosyl recognition marker required for receptor recognition, we covalently coupled purified receptor to an agarose gel bead support for affinity chromatography of phosphorylated, high mannose-type oligosaccharides isolated from fibroblast secretions radiolabeled with [2 3H]mannose. Neutral oligosaccharides and oligosaccharides containing one or two phosphates in phosphodiester linkage were not retained by the receptor column. By contrast, oligosaccharides bearing one phosphomonoester moiety were retarded on the column; those bearing two phosphomonoesters were bound to the column and were eluted with 10 mM mannose 6-phosphate. The binding of the oligosaccharides to the immobilized receptor correlates with their ability to be pinocytosed by fibroblasts and shows that the preferred recognition marker for the phosphomannosyl-enzyme receptor is a high mannose-type oligosaccharide chain bearing two uncovered phosphomannosyl groups. PMID- 6286660 TI - Analysis of sequence microheterogeneity among zein messenger RNAs. AB - We have synthesized cDNA clones for maize zein proteins using mRNAs purified from developing endosperm. Analysis of these clones by in vitro translation of hybrid selected mRNAs suggested differences in sequence homology among the mRNAs for the different molecular weight zein polypeptides. These differences were also apparent in restriction maps of clones corresponding to the Mr = 22,000, 19,000, and 15,000 zeins. Using radioactive cDNA inserts as probes, we measured the extent of sequence homology among zein clones with a sensitive dot hybridization procedure. By this analysis, it was possible to distinguish clones corresponding to the different molecular weight zeins at low (Tm - 49 degrees C) to moderate (Tm - 35 degrees C) criteria, while under more stringent conditions (Tm - 20 degrees C), distinctions could be made between zein sequences within a molecular weight group. This analysis distinguish three different mRNAs for each of the Mr = 22,000 and Mr = 19,000 zeins, but only one was detected for the Mr = 15,000 zein. Comparison of the nucleotide sequences of clones for the Mr = 22,000 and Mr = 19,000 zeins showed about 60% homology throughout the coding regions. This analysis also revealed the presence of short repetitive nucleotide sequences corresponding to tandem repeats of approximately 20 amino acids in both groups of proteins. PMID- 6286661 TI - 1H electron-nuclear double resonance of cobalt hemoglobin. AB - Proton 1H electron-nuclear double resonance spectra were obtained from cobalt substituted hemoglobin A. For oxy cobalt-substituted hemoglobin (CoHb), two major couplings were found. One is exchangeable by deuterium and has a coupling of approximately 5.6 MHz which is assigned to the N epsilon 2-proton of the distal histidine (E7), interacting with the ligand. The other splitting (approximately 2.4-3.2 MHz) is attributed to a methyl-proton on Val (E11). No coupling of protons on the proximal side is resolved. Deoxy CoHb exhibits only one prominent interaction (approximately 1.3 MHz) which is assigned to one of the protons on the proximal histidine (F8). The influence of the distal amino acids His (E7) and Val (E11) on the ligand affinity in oxy CoHb is discussed. PMID- 6286662 TI - Modification of the catalytic subunit of bovine heart cAMP-dependent protein kinase with affinity labels related to peptide substrates. AB - The modification and concomitant inactivation of the catalytic subunit of bovine heart cAMP-dependent protein kinase with affinity analogs of peptide substrates potentially capable of undergoing disulfide interchange with enzyme-bound sulfhydryl groups have been used to probe the active site associated with peptide binding. The regeneration of catalytic activity on treatment of the modified enzymes with dithiothreitol and the observation that prior reaction with 5,5' dithiobis-(2-nitrobenzoic acid) blocks the modification of the kinase by these reagents are consistent with the proposal that only thiol residues are reacting. The affinity analog Leu-Arg-Arg-Ala-Cys(3-nitro-2-pyridinesulfenyl)-Leu-Gly, 1, and the closely related peptide AcLeu-Arg-Arg-Ala-Cys(3-nitro-2-pyridinesulfenyl) Leu-Gly-OEt, 3, react with a single sulfhydryl as shown by the stoichiometry of the release of the 3-nitro-2-pyridinesulfenyl group and the amount of label incorporated in the enzyme when the radioactively labeled peptide analog of 3 (peptide 4) is employed as the modifying agent. The kinetics of the reaction of 1 with 4.3 microM catalytic subunit was monophasic (employing substrate in excess conditions), yielding an apparent value of KI of approximately 40 microM and a k2 value of approximately 0.25 s-1. The low value of the observed KI, together with the observation that protein kinase substrates inhibit the modification reactions, suggest strongly that the cysteine residue undergoing reaction is in the vicinity of the active site. By trypsin-catalyzed degradation and identification of the peptide segment modified by covalent attachment of the peptide portion of the radioactive analog 4, the single cysteine modified was identified as cysteine-198. PMID- 6286663 TI - Identification of a cardiac carnitine binding protein. AB - The existence of a cardiac carnitine binding protein was demonstrated using an in vitro binding assay. The binding activity was solubilized with Triton X-100 from the pellet following a 59,000 X g centrifugation of rat ventricular homogenates. Preincubation at temperatures above 40 degrees C or treatment with pronase significantly reduced the binding activity, suggesting that the activity was that of a protein. Cell fractionation studies suggested that the cardiac carnitine binding protein was associated with the plasma membrane fraction and that its activity was distinct from carnitine palmitoyl-transferase, carnitine acetyltransferase, and carnitine translocase. Optimal binding required 60 min of incubation at 25 degrees C. The binding of carnitine to the cardiac carnitine binding protein was saturable, and a dissociation constant of 0.7 microM for DL carnitine was measured. L-Carnitine competed with DL-[methyl-3H]carnitine for competition by D-carnitine was much less effective. Binding was significantly inhibited when N-ethylmaleimide, iodoacetic acid, or mercuric chloride was present. Once DL-[3H]carnitine was bound to the cardiac carnitine binding protein, radioactivity could be dissociated by a variety of mild treatments including dialysis, overnight incubation at 4 degrees C, and application to a gel filtration column. PMID- 6286664 TI - The role of nonspecific hydrophobic interactions in the biological activity of N epsilon-acyl derivatives of glucagon. Studies of conformation, receptor binding, and adenylate cyclase activation. AB - Glucagon was acylated at position 12 using conditions favoring reaction with the epsilon-amino group of lysine. The N epsilon-acetyl, N epsilon-hexanoyl, and N epsilon-decanoyl derivatives were prepared and purified. Secondary structure as measured by circular dichroism was lower in all derivatives than in glucagon, both in 95% methanol and in 25 mM sodium dodecyl sulfate at pH 2 and pH 12. N epsilon-Acetyl glucagon was less active than the native hormone in a radioreceptor assay and higher concentrations of this derivative were required to stimulate the adenylate cyclase activity of rat liver plasma membranes. The maximal extent of cyclase activation by this derivative was less than that found with the native hormone. N epsilon-Hexanoyl glucagon and N epsilon-decanoyl glucagon had greater activity than N epsilon-acetyl glucagon in receptor binding as well as in adenylate cyclase activation, although these two derivatives were not as active as the native hormone. N epsilon-hexanoyl glucagon and N epsilon decanoyl glucagon were more potent in receptor binding than in adenylate cyclase activation. From these results it appears that the positive charge of the epsilon amino groups may have a specific role in obtaining maximal biological activity, while the acyl groups contribute to the nonspecific hydrophobic interactions between the hormone and its receptor. In addition, a possible relationship between stabilization of the amphipathic helix in solution and the activity of these and other N epsilon-derivatives of glucagon is discussed. PMID- 6286665 TI - Ca2+-dependent interaction of 5-dimethylaminonaphthalene-1-sulfonyl-calmodulin with cyclic nucleotide phosphodiesterase, calcineurin, and troponin I. PMID- 6286666 TI - AMP deaminase as a control system of glycolysis in yeast. Mechanism of the inhibition of glycolysis by fatty acid and citrate. AB - The role of fatty acid and citrate on the interaction of the AMP deaminase (EC 3.5.4.6) reaction with glycolysis was investigated using permeabilized yeast cells. (a) Linolenate and citrate inhibited glycolytic flux and the recovery of the adenylate energy charge; however, linolenate remarkably retarded the depletion of the total adenylate pool, which was not at all affected by the addition of citrate. (b) Linolenate inhibited AMP deaminase activity in situ, resulting in the subsequent decrease in ammonium production, which reduced the activity of 6-phosphofructokinase (EC 2.7.1.11), whereas linolenate itself had no ability to inhibit the phosphofructokinase activity in the presence of excess ammonium concentration. (c) Citrate inhibited the activity of phosphofructokinase in situ in the presence and absence of ammonium ion, followed by an inhibition of glycolysis; however, AMP deaminase activity was not inhibited by citrate. The inhibition of glycolysis by fatty acids can be accounted for by the lowered activity of phosphofructokinase as a result of the decreased level of ammonium ion through the inhibition of the AMP deaminase reaction by these ligands, whereas the effect of citrate on glycolysis is a direct inhibition of phosphofructokinase without affecting the activity of AMP deaminase. Fatty acid and citrate, a principal metabolic product of fatty acid oxidation, can be responsible for the control of glycolysis in two different manners. PMID- 6286667 TI - Conformational change accompanying transition of ADP-sensitive phosphoenzyme to potassium-sensitive phosphoenzyme of (Na+,K+)-ATPase modified with N-[p-(2 benzimidazolyl)phenyl]maleimide. AB - The addition of Mg2+ or ATP to (Na+,K+)-ATPase (EC 3.6.1.3) of pig kidney modified with a sulfhydryl fluorescent reagent N-[p-(2 benzimidazolyl)phenyl]maleimide simply reduced fluorescence in the presence of Na+; however, the addition of both ligands to the enzyme induced a reversible dynamic change. The direction of the change was dependent on the concentration of Na+ present. These dynamic changes in fluorescence intensity both in the presence of low and high concentrations of Na+ can be repeated by the re-addition of ATP but not by ADP. Addition of ouabain under the former condition stabilized the fluorescence at the highest level, but the addition of ouabain under the latter condition increased the fluorescence from the lowest to the highest level. The phosphoenzyme formed under the former condition was sensitive to K+ and insensitive to ADP while the phosphoenzyme formed under the latter condition was sensitive to ADP and insensitive to K+. The data indicate that the positive and negative fluorescence changes were induced by the formation of K+-sensitive phosphoenzyme and ADP-sensitive phosphoenzyme, respectively. N-Ethylmaleimide treatment partially inhibited the positive change without affecting the negative change. These data also indicate that the transition of ADP-sensitive phosphoenzyme to K+-sensitive phosphoenzyme accompanied the largest fluorescence intensity change which was examined during the hydrolysis of ATP. The data obtained from the tryptophan fluorescence of both the native and the modified enzyme suggest that the micro-environments of the tryptophan and the sulfhydryl residues are similar in the state of K+-sensitive phosphoenzyme but different in the state of ADP-sensitive phosphoenzyme. PMID- 6286668 TI - Linkage arrangement of human placental lactogen and growth hormone genes. AB - Human placental lactogen (hPL) and growth hormone (hGH) are two hormones thought to have evolved from a common ancestral gene (along with prolactin), yet they have quite different functions and specificities. The nucleic acid sequences of the respective cDNAs of the two genes share considerable homology, as well as the existence of multiple forms of each gene within the genome. In this study we report on the linkage arrangement of several genes from this group. Two hPL-like genes as well as an hGH gene are shown to be linked within a 38-kilobase pair region of DNA. Linkage between a variant hGH gene and an hPL gene is also shown. The orientation and structural organization of these genes was previously established using 5'- and 3'-specific probes from a placental lactogen cDNA clone and detailed restriction endonuclease mapping. Restriction fragments from the overlapping clones were verified by comparison to digests of high molecular weight genomic DNA. In addition, the location of a specific class of repetitive DNA sequences, the Alu family, was mapped on these clones using the recombinant clone BLUR 8. All members of this multigene family have Alu repeat sequences either immediately flanking their 3' or 5' untranslated regions or within their intervening sequences. PMID- 6286670 TI - Different classes of nucleotide binding sites in the (Na+ + K+)-ATPase studied by affinity labeling and nucleotide-dependent SH-group modifications. AB - The ATP analog 6-[(3-carboxy-4-nitrophenyl)thiol]-9-beta-D-ribofuranosylpurine 5' triphosphate (Nbs6ITP) is slowly hydrolyzed at pH 7.4 by the (Na+ + K+)-ATPase, whereas it binds covalently at pH 8.5 and inhibits the enzyme irreversibly. Time courses of irreversible inhibition could only be fitted to a model in which the enzyme can exist in two slowly interchangeable states, one of which is enzymatically active and binds Nbs6ITP first reversibly and then covalently. Arguments that the covalent binding occurs at a low affinity nucleotide binding site are: (a) similarity of the Ki Nbs6ITP for the reversible and the irreversible inhibition and of K0.5 for ATP protection; (b) stoichiometry of covalent Nbs6ITP binding per alpha subunit of 0.8; and (c) change of complex substrate dependence of the enzyme to a Michaelis-Menten type after Nbs6ITP modification. This change in kinetics and the finding that the Nbs6ITP inactivation at a low affinity nucleotide binding site is increased by micromolar ADP concentrations indicates that the (Na+ + K+)-ATPase contains two different nucleotide binding sites. Since studies of nucleotide effects on enzyme inactivation by 5,5'-dithiobis(2-nitrobenzoic acid) did not confirm the hypothesis of an SH-group in a nucleotide binding site, Nbs6ITP may bind to another functional group, e.g. to an OH-group of tyrosine. PMID- 6286669 TI - The insulin-directed phosphorylation site on ATP-citrate lyase is identical with the site phosphorylated by the cAMP-dependent protein kinase in vitro. AB - 32P-labeled ATP-citrate lyase isolated from 32P-labeled hepatocytes treated with insulin contained 1.6-1.8-fold greater 32P-radioactivity per mg protein than control enzyme. Both enzyme preparations were digested in parallel with trypsin until 94% of all 32P-radioactivity was rendered acid soluble. Quantitative high performance liquid chromatographic peptide mapping of the tryptic digests revealed a principal 32P-peptide which accounted for at least 80% of the insulin induced increment in 32P-radioactivity of native lyase. This peptide was purified, sequenced, and the site of 32P-phosphorylation assigned by two methods: electrophoresis (pH 6.5) of residual peptide after each step of Edman degradation and solid phase sequencing. The site of insulin-directed phosphorylation of ATP citrate lyase (Thr-Ala-Ser(32P)-Phe-Ser-Glu-Ser-Arg) is the same as that directed by glucagon, and, in turn, identical with that phosphorylated by the cAMP dependent protein kinase in vitro. PMID- 6286671 TI - Kinetics and mechanism of the reduction of ferricytochrome c by the superoxide anion. AB - The temperature and pH dependence of the reaction of the superoxide radical anion with ferricytochrome c have been measured using the pulse-radiolysis technique. The temperature dependence of the reaction at low ionic strength yields an activation energy of 31 +/- 5 kJ/mol as compared to 14 +/- 3 kJ/mol for the reaction of CO2.(-) under the same conditions. The pH dependence fits the single pK'a of ferricytochrome c of 9.1. The bimolecular rate constant for the reaction of the superoxide anion with ferricytochrome c at pH 7.8, 21 +/- 2 degrees C, in the presence of 50 mM phosphate and 0.1 mM EDTA is (2.6 +/- 0.1) X 10(5) M-1 s-1. Using this value, 1 unit of superoxide dismutase activity (McCord, J. M., and Fridovich, I. (1969) J. Biol. Chem. 244, 6049-6055) is calculated to be 3.6 +/- 0.3 pmol of enzyme if the assay is performed in a total volume of 3.0 ml. Copper ions reduce the yield of the reaction of ferricytochrome c with CO2.(-). The reactivities of native and singly modified 4-carboxy-2,4-dinitrophenyllysine cytochromes c towards the superoxide anion radical are in the order native greater than 4-carboxy-2,4-dinitrophenyllysine 60 greater than lysine 13 greater than lysine 87 greater than lysine 27 greater than lysine 86 greater than lysine 72, indicating that electron transfer takes place at or close to the solvent accessible heme edge. The mechanism of the reaction is discussed in terms of the approach of superoxide anion radicals to the heme edge and the available molecular orbitals of both heme and free radicals. PMID- 6286672 TI - Phosphatidylcholine-promoted interaction of the catalytic and regulatory proteins of adenylate cyclase. AB - The catalytic protein of rabbit hepatic adenylate cyclase, after chromatographic separation from the GTP-binding regulatory protein (G/F) (Ross, E. M. (1981) J. Biol. Chem. 256, 1949-1953), is essentially free of endogenous phospholipids. This preparation is active in the presence of Mn2+ and is markedly stimulated by forskolin, but it is stimulated only slightly by the addition of purified G/F plus an activator (GTP gamma S or NaF). The ability of activator-liganded G/F to stimulate the activity of the resolved catalyst is increased up to 8-fold by the addition of either dimyristoylphosphatidylcholine (0.3-1.5 mM optimal concentration) or several other phosphatidylcholines. Phosphatidylcholine stabilizes the catalyst to denaturation but has little effect on its basal activity or on its stimulation by Mn2+ or forskolin. It also had no stimulating effect on the activation of G/F by GTP gamma S. These data are interpreted as showing that phosphatidylcholine promotes or is required for the stimulatory interaction of activator-liganded G/F with the catalytic protein of adenylate cyclase. Lubrol 12A9, Triton X-100, cholate, lysophosphatidylcholine, digitonin, and phosphatidylserine could not substitute for phosphatidylcholine. The detergents inhibited stimulation by liganded G/F even in the presence of phosphatidylcholine. Removal of cholate from a mixture of soluble catalytic protein and phosphatidylcholine by dialysis and sucrose density gradient centrifugation caused the binding of catalytic protein to large unilamellar vesicles. This preparation was further reconstituted with increasing amounts of G/F to yield vesicles with varied G/F:catalyst ratios and similarly varied responses to G/F-mediated activating ligands. PMID- 6286673 TI - Studies of the Ca2+ transport mechanism of human erythrocyte inside-out membrane vesicles. Evidence for the development of a positive interior membrane potential. AB - Previous observations on the effects of permeant anions on ATP-dependent calcium transport in inside-out vesicles prepared from human erythrocytes suggested that the calcium pump is electrogenic, generating a positive interior membrane potential. The present work demonstrates the development of a positive interior membrane potential across inside-out vesicle membranes during calcium transport in the absence of permeant anions. Several membrane potential probes, 1-anilino-8 naphthalenesulfonate, 3,3'-dipropylthiodicarbocyanine iodide, and an electron paramagnetic resonant triphenylphosphonium derivative, provide qualitative evidence for the development of a membrane potential. Moreover, a number of parallels are observed between the changes in the membrane potential measured by the probes and calcium transport. These include enhancement by calmodulin, time course of change, similar kinetic properties, and the requirement for intact vesicle membranes. Quantitative measurements of the membrane potential shows a positive interior membrane potential of 26-37 mV using radiolabeled permeant anion distribution and 38-57 mV using 3,3'-dipropylthiodicarbocyanine iodide fluorescence changes. These membrane potentials are of a similar magnitude to those reported for the sarcoplasmic reticulum calcium pump (Zimniak, P., and Racker, E. (1978). J. Biol. Chem. 253, 4631-4637). PMID- 6286674 TI - Lateral diffusion of ubiquinone during electron transfer in phospholipid- and ubiquinone-enriched mitochondrial membranes. AB - After fusion of small unilamellar phospholipid liposomes with mitochondrial inner membranes, the rate of electron transfer between membrane dehydrogenases and cytochrome c decreases as the average distance between integral membrane proteins increases, suggesting that electron transfer is mediated through a diffusional process in the membrane plane (Schneider, H., Lemasters, J. J., Hochli, M., and Hackenbrock, C. R. (1980)., J. Biol. Chem. 255, 3748-3756). The role of ubiquinone in this process was evaluated by fusing liposomes containing ubiquinone-10 or ubiquinone-6, with inner membranes. In control membranes enriched with phospholipid only, ubiquinol-cytochrome c reductase and NADH- and succinate-cytochrome c reductase activities decreased proportionally to the increase in bilayer lipid. These decreases were restored substantially in phospholipid plus ubiquinone-supplemented membranes. The degree to which restoration occurred was dependent upon the length of the isoprenoid side chain of the ubiquinone with the shorter chain length ubiquinone-6, always giving greater restoration than ubiquinone-10. It is concluded that electron transfer between flavin-linked dehydrogenases (Complexes I and II) and cytochrome bc1 (Complex III) occurs by independent, lateral diffusion of ubiquinone as well as independent, lateral diffusion of ubiquinone as well as the protein complexes within the plane of the membrane. PMID- 6286675 TI - Radiation inactivation studies of the renal brush-border membrane phlorizin binding protein. AB - Renal brush-border membrane vesicles were irradiated in the frozen state with a high energy electron beam. The integral membrane proteins, alkaline phosphatase and 5'-nucleotidase, each showed a single exponential loss of activity with radiation dose, indicating target sizes of 67,000 and 58,000 daltons, respectively. Inactivation of sodium-dependent phlorizin binding to the brush border membrane D-glucose transporter was more complex. One-half of the phlorizin binding sites were lost after even the smallest doses of radiation suggestive of large functional units (greater than 4 X 10(6) daltons) for a subpopulation of phlorizin binding proteins. The remaining sites behaved as a single radiation target of 110,000 +/- 8,000 daltons and retained the kinetic characteristics commonly associated with phlorizin binding to the glucose transporter. Thus, the data are consistent with the assignment of a molecular weight of 110,000 to the phlorizin binding moiety of the brush-border membrane D-glucose transport protein. PMID- 6286676 TI - Proteolytic solubilization of adenylate cyclase from membranes deficient in regulatory component. Properties of the solubilized enzyme. PMID- 6286677 TI - Sodium channel, sodium pump, and sodium-calcium exchange activities in synaptosomal plasma membrane vesicles. AB - Two mechanisms of Na+ influx have been observed using synaptosomal plasma membrane vesicles purified by density gradient centrifugation from a synaptosomal hypotonic lysate. First, a 5-fold increase in uptake over basal Na+ entry occurs with 50 microM veratridine. The veratridine-dependent Na+ uptake is partially inhibited by 2 microM tetrodotoxin with an apparent time dependency of action (half-maximal inhibition in approximately 20 min). Second, a larger Na+ accumulation (approximately 15-fold above basal) was observed with 2.5 mM ATP, this effect being dependent on internal K+ loading of vesicles although inhibited by high external K+. The two uptake processes are believed to represent operation of the plasma membrane voltage-sensitive Na+ channel, and the Na+-pumping (Na+ + K+)-ATPase, respectively. Both Na+ flux mechanisms appear to operate in a single population of vesicles since opening of the Na+ channel with veratridine diminishes the ATP-dependent accumulation of Na+ by over 75%. An inverted orientation of the plasma membrane vesicles is likely to account for the functioning of the ATP-dependent Na+ pump and may also account for the low sensitivity and time dependency of the inhibitory action of tetrodotoxin on Na+ channel-opening. Na+ accumulated by the Na+ pump was rapidly effluxed by 10 mM external Ca2+ via the Na+-Ca2+ exchange mechanism which (together with an ATP dependent Ca2+-accumulating mechanism) was recently characterized in the vesicles (Gill, D. L., Grollman, E. F., and Kohn L. D. (1981) J. Biol. Chem. 256, 184 192). This result, together with the observed inhibition of Ca2+ influx via (Na+ Ca2+)-exchange due to veratridine-mediated Na+ flux, strongly suggests that the Na+ pump, Na+ channel, and both Ca2+ transport mechanisms function in a single population of inverted plasma membrane vesicles. PMID- 6286678 TI - Transcriptional properties of chick embryonic erythroid nuclei in vitro. AB - Chick embryo red blood cells express the embryonic globin and the rRNA genes before 5 days of development but only the adult globin genes and no rRNA after 12 days of development. We have isolated nuclei from the red blood cells of these developmental stages and allowed them to transcribe in vitro. We have analyzed the overall transcriptional properties of these nuclei, the overall activities of RNA polymerases I and II and the sequence-specific activity of RNA polymerase II in the beta-globin domain using cloned genomic hybridization probes. Among our findings are the following. 1) Erythroid nuclei of 5-day embryos are more transcriptionally active than those at 12 days. 2) RNA polymerase I is a very active at 5 days but is off by 12 days. 3) A template-independent activity which yields labeled RNA is present in the red cell nuclei of 12-day but not 5-day embryos. 4) Between 5 and 12 days of development transcriptional modulation delineates embryonic and adult beta-globin domains. 5) These domains exceed the boundaries of the genes themselves by several kilobases. 6) All transcripts which hybridize to sequences in the beta-globin gene region, including repeat sequence transcripts, are transcribed by RNA polymerase II. PMID- 6286679 TI - The chicken beta globin gene region. Delineation of transcription units and developmental regulation of interspersed DNA repeats. PMID- 6286680 TI - Point mutagenesis of the ovalbumin gene promoter sequence and its effect on in vitro transcription. AB - The role of the eucaryotic T-A-T-A box (Hogness box) homology sequence located approximately 30 base pairs upstream from the RNA initiation site has been further examined by oligodeoxynucleotide-directed site-specific mutagenesis. A method was employed which allows insertion of nucleotide-specific mutations into virtually any double-stranded, recombinant plasmid DNA. A synthetic mixed oligonucleotide, bearing defined multiple nucleotide substitutions at a single site, was used both as a specific mutagen during primed DNA repair synthesis in vitro, as well as a highly sensitive hybridization probe for the identification of the mutated cloned DNA. Using this methodology, an A leads to G transition mutation was introduced into the second position of the ovalbumin gene T-[A]-T-A box, and the effect of modifying this highly conserved nucleotide on the expression of the mutant DNA was analyzed in a cell-free transcription system. Comparison of two allelic ovalbumin genes, slightly divergent upstream from the TATA box, resulted in identical in vitro transcription efficiencies. While correct initiation of ovalbumin RNA transcripts was not affected, the efficiency of gene expression using the mutant template, compared to either the corresponding wild-type sequence or the allelic gene, was markedly reduced. These results suggest that it is the T-A-T-A box sequence which plays a role in the efficient initiation of RNA transcription in vitro and further supports the implication that this region may serve a promoter-related function in eucaryotic transcription. PMID- 6286681 TI - Purification and characterization of the gamma regulatory subunit of the cyclic GMP phosphodiesterase from retinal rod outer segments. AB - Retinal rod outer segments contain a phosphodiesterase specific for cyclic GMP. This enzyme is virtually inactive in the dark. Photoexcitation of rhodopsin results in the formation of hundreds of molecules of GTP-transducin, which in turn activate many molecules of phosphodiesterase. The phosphodiesterase is also known to be activated by the proteolytic action of trypsin. We have investigated the nature of the inhibitory constraint on the catalytic activity of the phosphodiesterase in the dark state. Phosphodiesterase purified by hexylagarose chromatography followed by gel filtration high pressure liquid chromatography consists of three kinds of subunits: alpha (88 kilodaltons), beta (84 kilodaltons), and gamma (11 kilodaltons). Three lines of evidence show that the phosphodiesterase in the dark state is inhibited by its gamma subunit. First, inhibitor activity copurifies with the catalytic activity of this enzyme. Second, trypsin degrades the gamma subunit, resulting in a concomitant increase in catalytic activity. The high pressure liquid chromatography elution position of trypsin-activated phosphodiesterase suggests that it is an alpha beta complex. Third, nearly all of the catalytic activity of trypsin-activated phosphodiesterase can be inhibited by the addition of gamma subunit purified either by heat treatment or by gel filtration at pH 2.1. The addition of gamma subunit to trypsin-activated phosphodiesterase decreases its Vmax from 1.2 mmol of cyclic GMP hydrolyzed/min/mg to less than 1% of this value with relatively little change in the value of Km. The gamma subunit has high affinity for trypsin activated phosphodiesterase. The dissociation constant of this complex is 0.13 nM. These experiments show that the phosphodiesterase in the dark state has very little catalytic activity because of the inhibitory constraint imposed by its gamma subunit. PMID- 6286682 TI - In vitro transcription of the supB-E tRNA operon of Escherichia coli. Characterization of transcription products. AB - The seven tRNA genes clustered in the supB-E region of the Escherichia coli chromosome were transcribed in vitro with purified RNA polymerase, using a restriction fragment from lambda psu degrees 2, a transducing phage carrying the chromosome region, as template. A single major transcript was synthesized, which was about 770 nucleotides long and contained all seven tRNA sequences. The terminal sequences of the transcript were determined and mapped on the DNA sequence of the supB-E region previously determined. The transcription start site is seven base pairs downstream from the Pribnow box sequence, as expected from the DNA sequence analysis and consistent with the findings on the trimeric tRNA precursor (pppG--tRNAMETM-tRNALeu-tRNAGln1) which was detected in an RNase P mutant and shown to be coded for by the supB-E region. Cleavage of the restriction fragment at the -35 region with another restriction endonuclease abolished the template activity of the fragment. Transcription of the supB-E tRNA operon was relatively unaffected by the presence of rho factor. Transcription termination occurs within a region of three bases between positions 770 and 772 from the transcription start site. Immediately upstream from the termination sites, there is a region of 26 nucleotides that could form a stem structure, thereby consistent with the general feature of rho-independent termination sites. PMID- 6286683 TI - The predominant secreted protein of transformed murine fibroblasts carries the lysosomal mannose 6-phosphate recognition marker. AB - We have found that the major excreted protein (MEP) of transformed mouse fibroblasts, a phosphoglycoprotein of Mr = 35,000, carries the mannose 6 phosphate recognition marker. MEP secreted by Kirsten virus-transformed NIH 3T3 cells binds to a purified preparation of lysosomal enzyme phosphomannosyl receptor, and this binding is specifically inhibited by mannose 6-phosphate. 32Pi introduced into MEP by metabolic labeling of intact cells is exclusively associated with asparagine-linked oligosaccharides as indicated by sensitivity to endohexosaminidase H. Labeling studies utilizing [2-3H]mannose indicate that approximately one-fifth of the mannose residues of MEP are phosphorylated. Comparative studies of the synthesis, secretion, and uptake of MEP and the lysosomal enzyme beta-galactosidase indicate that MEP made by Kirsten virus transformed NIH 3T3 cells is not handled in the same manner as are other lysosomal enzymes. MEP may be an unusual lysosomal protein, or both. PMID- 6286685 TI - Scanning electron microscopic study of the hydrolytic degradation of poly(glycolic acid) suture. AB - This article reports the morphological observations on the surface changes of poly-(glycolic acid) sutures which have been exposed to various dosages of gamma irradiation (0, 2.5, 5.0, 10, 20 and 40 Mrad) and duration of immersion (0, 7, 14, 28, 48, 60, and 90 days) in a physiological saline buffer. The most important gross morphological characteristics of PGA suture hydrolytic degradation is the formation of surface cracks on the filaments. The regularity of the surface cracks increased with an increase in the gamma irradiation and the duration of hydrolysis. Surface cracks were not observed in irradiated sutures that had not been subjected to hydrolytic degradation. The arrangement of the surface cracks, their orientation on the filaments, and the direction of crack propagation provide very useful information for depicting the mechanism of hydrolytic degradation in this class of fibrous material. The microfibrillar model of fiber structure has been used as the basis for the proposed degradation mechanism of PGA in vitro. It is believed that hydrolysis occurs initially in the amorphous regions sandwiched between two crystalline zones, as tie-chain segments, free chain ends, and chain folds in these regions degrade into fragments. As degradation proceeds, the size of the fragments reaches the stage at which they can be dissolved into the buffer medium. This dissolution removes the fragments from the amorphous regions, and surface cracks appeared. PMID- 6286684 TI - Isolation of the yeast nuclear gene encoding the mitochondrial protein, cytochrome c peroxidase. PMID- 6286686 TI - Adsorbed behavior of spin-labeled poly(methyl methacrylate) on human tooth and hydroxyapatite studied by electron spin resonance. PMID- 6286687 TI - The distal femoral defect: technetium-99m pyrophosphate bone scan results. AB - To determine the scintigraphic findings in children with distal femoral defects (or subperiosteal desmoids) the authors retrospectively examined 94 sets of radiographs and technetium-99m pyrophosphate bone scans of 54 children. Twenty four examples of distal femoral defects were identified in 19 children. Only two children demonstrate focally increased uptake of radionuclide in the distal femur. The positive bone scan was caused by osteomyelitis in one of these patients and by metastatic lymphoma in the other. None of the 17 other children with distal femoral defects on radiography had a corresponding focal increase in uptake of radionuclide. The bone scan appears to be a useful method for confirming that a distal femoral defect is benign and, when positive, indicates the presence of other disease. PMID- 6286688 TI - Guinea pig adrenocortical cells: in vitro characterization of separated zonal cell types. AB - This paper reports a quick, relatively simple and reproducible technique for obtaining populations of zona fasciculata and zona glomerulosa cells up to 80-90% pure, which can be maintained in vitro for study of adrenocortical cell function. Isolated guinea pig adrenocortical cells were separated on a 1-28% bovine serum albumin/Ca++, Mg++-free buffer gradient (wt/vol at 4% increments) using equilibrium density centrifugation (570 g, 30 min). Over 60% of the 8 x 10(5) viable cells/adrenal obtained in the total isolate were recovered after separation. 80% of the zona glomerulosa cells were found in the lower three bands of the gradient. 78% of the zona fasciculata cells were found in the top three bands. Of the cells in the first two bands, 78-91% were zona fasciculata cells, whereas of the cells in the bottom two bands 92-95% were zona glomerulosa cells. The cells retained the morphological characteristics of cells in situ and could be maintained in vitro for periods up to 11 d. They produced a wide variety of steroids, cortisol, corticosterone, aldosterone, 11-beta-hydroxyandrostenedione, deoxycortisol, deoxycorticosterone, cortisone, 18-hydroxycorticosterone, and a product tentatively identified as dehydroepiandrosterone, and they responded to ACTH in a dose-responsive manner with enhanced levels of steroid output. Zona glomerulosa-enriched populations differed from zona fasciculata-enriched populations in their abundant production of aldosterone and in the pattern of steroid production. None of the cultures responded to angiotensin II (100 pg/ml) with increased steroid production. PMID- 6286689 TI - Nonequilibrium kinetics of a cyclic GMP-binding protein in Dictyostelium discoideum. AB - Chemoattractants added to cells of the cellular slime mold dictyostelium discoideum induce a transient elevation of cyclic GMP levels, with a maximum at 10 s and a recovery of basal levels at approximately 25 s after stimulation. We analyzed the kinetics of an intracellular cGMP binding protein in vitro and in vivo. The cyclic GMP binding protein in vitro at 0 degrees C can be described by its kinetic constants K(1)=2.5 x 10(6) M(- 1)s(-1), k(-1)=3.5 x 10(-3)s(-1), K(d)=1.4 x 10(-9) M, and 3,000 binding sites/cell. In computer simulation experiments the occupancy of the cGMP binding protein was calculated under nonequilibrium conditions by making use of the kinetic constants of the binding protein and of the shape of the cGMP accumulations. These experiments show that under nonequilibrium conditions by making use of the kinetic constants of the binding protein and the shape of the cGMP accumulations. These experiments show that under nonequilibrium conditions the affinity of the binding protein for cGMP is determined by the rate constant of association (k(1)) and not by the dissociation constant (k(d)). Experiments in vivo were performed by stimulation of aggregative cells with the chemoattractant cAMP, which results in a transient cGMP accumulation. At different times after stimulation with various cAMP concentrations, the cells were homogenized and immediately thereafter the number of binding proteins which were not occupied with native cGMP were determined. The results of these experiments in vivo are in good agreement with the results of the computer experiments. This may indicate that: (a) The cGMP binding protein in vivo at 22 degrees C can be described by its kinetic constants: K(1)=4x10(6)M( 1)s(-1) and K(-1)=6x10(-3)s(-1). (b) Binding the cGMP to its binding protein is transient with a maximum at about 20-30 s after chemotactic stimulation, followed by a decay to basal levels, with a half-life of approximately 2 min. (c) The cGMP to its binding proteins get half maximally occupied at a cGMP accumulation of delta[cGMP](10)=2x10(-8) M, which corresponds to an extracellular stimulation of aggregative cells by 10(-10) M cAMP. (d) Since the mean basal cGMP concentration is approximately 2x10(-7) M, the small increase of cGMP cannot be detected accurately. Therefore the absence of a measurable cGMP accumulation does not argue against a cGMP function. (e) There may exist two compartments of cGMP: one contains almost all the cGMP of unstimulated cells, and the other contains cGMP binding proteins and the cGMP which accumulates after chemotactic stimulation. (f) From the kinetics of binding, the cellular responses to the chemoattractant can be divided into two classes: responses which can be mediated by this binding protein (such as light scattering, proton extrusion, PDE induction, and chemotaxis) and responses which cannot be (solely) mediated by this binding protein such as rlay, refractoriness, phospholipids methylation, and protein methylation. PMID- 6286690 TI - Loss of covalently labeled glycoproteins and glycolipids from the surface of newly transformed schistosomula of Schistosoma mansoni. AB - Schistosomula of Schistosoma mansoni were labeled by oxidation with galactose oxidase or with periodate followed by reduction with NaB3H4 to study the loss of the surface membrane of these parasites in vitro. Grain counts of light microscope autoradiographs (LMARG) of radiolabeled schistosomula show that both galactose oxidase and periodate specifically label the surface of the organisms. Galactose oxidase labels 11 glycoproteins on the surface of skin and mechanical schistosomula, ranging in apparent molecular weight from 17,000 to greater than 105,000. These glycoproteins are lost from the surface of schistosomula with a halftime of 10-15 h in culture in defined medium. Most of these glycoproteins appear to be shed intact from the surface of the schistosomula rather than endocytosed and degraded, because greater than 50% of each of the lost proteins can be recovered by trichloroacetic acid precipitation of the culture medium and because there is no internalization of the radiolabels into cultured schistosomula examined by LMARG. In addition to glycoproteins, periodate labels at least seven glycolipids on the surface of mechanical schistosomula. After culture for 15 h, more than half of each of these periodate-labeled proteins and lipids are lost from the schistosomula, and their abundance relative to each other remains similar to that of freshly labeled organisms. Since both proteins and lipids are lost from the surface of the schistosomula at the same rate, we believe that we are observing a general loss of the parasite surface membrane. PMID- 6286692 TI - Modulations of Na+ transport during the cell cycle of neuroblastoma cells. PMID- 6286691 TI - Parathyroid hormone receptor in intact embryonic chicken bone: characterization and cellular localization. AB - The specific localization and the characterization of the parathyroid hormone (PTH) receptor in bone have been studied using 18-d embryonic chick calvariae and biologically active, electrolytically labeled [125I] bovine PTH(1-34). Binding was initiated by adding [125I]-bPTH(1-34) to bisected calvariae at 30 degrees C. Steady state binding was achieved at 90 min at which time 10 mg drg wt of calvaria specifically bound 17% of the added [125I]bPTH(1-34). Nonspecific binding in the presence of 244 nM unlabeled bPTH(1-34) was less than 2%. Insulin, glucagon, and calcitonin (1 microgram/ml) did not compete for PTH binding sites. Half-maximal inhibition of binding was achieved at concentrations of unlabeled bPTH(1-34) or bPTH(1-84) of about 10 nM. The range of concentration (2-100 nM) over which bPTH(1-34) and bPTH(1-84) stimulated cyclic 3'5'adenosine monophosphate (cAMP) production was similar to that which inhibited the binding of [125I]bPTH(1-34). Light microscope autoradiograms showed that grains were concentrated over cells (osteoblasts and progenitor cells) at the external surface of the calvariae and in trabeculae. In the presence of excess unlabeled PTH, labeling of control autoradiograms was reduced to near background levels. No labeling of osteocytes or osteoclasts was observed. At the electron microscopic level, grains were localized primarily over cell membranes. A quantitative analysis of grain distribution suggested that cellular internalization of PTH occurred. PMID- 6286693 TI - Binding of phorbol esters to high-affinity sites on murine fibroblastic cells elicits a mitogenic response. AB - The level of occupancy of a single class of high-affinity (3H)-phorbol 12, 13 dibutyrate (PDBu) binding sites (Kd = 26 nM) in quiescent Swiss 3T3 cells was correlated with the dose of PDBu required to stimulate rapid (increase in 86Rb uptake, decrease in epidermal growth factor receptor affinity) and long-term (induction of 2-deoxyglucose uptake, initiation of DNA synthesis) events of the proliferative response. Further, structural analogues of PDBu showed identical relative potencies in the inhibition of (3H)-PDBu binding and stimulation of DNA synthesis. Finally, prolonged (24-hour) incubation of Swiss 3T3 or whole mouse embryo fibroblasts with 400 nM PDBu led to a decrease in the number of (3H)-PDBu binding sites (down-regulation) with a parallel loss of rapid and long-term responses of the cells to PDBu (desensitization). These findings indicate that the high-affinity (3H)-PDBu binding sites mediate the mitogenic effects of phorbol esters in fibroblastic cells. PMID- 6286694 TI - Electron microscopic observations on the development and cytochemistry of the large granule complexes in chicken erythrocyte nuclei. AB - Large granule complexes are structures found in a small percentage of chicken erythrocyte nuclei when observed in ultra-thin sections in the electron microscope. They consist of an amorphous region associated with a number of large (approximately 30 min) granules. We have shown, by a novel use of phenylhydrazine to synchronize populations of chicken erythrocytes in vivo, that large granule complexes do not occur in the nuclei until the cells have reached one-third to one-half of their normal intravascular lifespan. The mature large granule complexes are formed by aggregation of pre-existing fibrillar, granular and amorphous material, and their presence is correlated with the presence of another ultrastructural feature of the nucleus, the so-called "filled cavities' in the chromatin. Digestion of ultra-thin sections of erythrocytes embedded in the hydrophilic resin glycol methacrylate (GMA) has shown that the major component of the amorphous region is a rather acidic protein that is not haemoglobin, the most abundant protein in the erythrocyte. The large granules also contain protein and, almost certainly, RNA. The problems encountered in reaching this conclusion have emphasized the lack of unambiguous cytochemical tests for use on ultra-thin sections. We have shown that the large granule complex differs in many respects from the nucleolus in the erythrocyte series, even though the two organelles have certain superficial similarities such as their overall dimensions and the presence of granular and fibrillar regions. The most likely function of the large granule complex is as a repository for material, including RNA, the processing of which has ceased in the inactivated erythrocyte nucleus. PMID- 6286695 TI - Effects of dexamethasone on antigen expression and virus production in avian sarcoma virus-transformed cells. AB - We studied the effects of dexamethasone on virus production and antigen expression in avian retrovirus-infected chick embryo fibroblast (CEF) cells. The presence of specific receptors for this hormone in both normal and infected CEF cells could only be demonstrated using cultures that were at or near confluence. Dexamethasone, while not toxic to the cells, exerted a growth-retarding influence when employed at concentrations of 10(-8) or 10(-9) M. These same concentrations of hormone were inhibitory to virus production, elaboration of plasminogen activator activity, and antigen expression as determined in a sensitized lymphocyte stimulation assay. In contrast, infected cells that had been treated with hormone displayed enhanced reactivity in a specific immunofluorescence test. PMID- 6286696 TI - Macrocyst development in Dictyostelium discoideum. I. Induction of synchronous development by giant cells and biochemical analysis. AB - In Dictyostelium discoideum, cytological and physiological studies on macrocyst formation revealed that this process consists of at least two steps: the production of giant cells, which are believed to be formed from the fusion of cells of two opposite mating types, and the subsequent induction of macrocyst development by the giant cells. The conditions that had been considered formerly to be required for macrocyst formation, such as darkness at the presence of two cells of complementary mating types in heterothallic strains, were actually required only for the production of the giant cells. Once giant cells are produced, the surrounding cells can aggregate and form macrocysts even in the light. Furthermore, it was demonstrated that giant cells can switch the developmental mode of the surrounding cells to macrocyst formation. That is, if a critical number of the isolated giant cells are introduced into a cell population of a single strain of NC4, which normally would produce only fruiting-bodies, macrocysts are formed instead. When in the presence of giant cells, the development of macrocysts may be initiated by starvation. Therefore, if all cells are made to starve simultaneously development begins and proceeds synchronously. Using this technique of synchronous development, the developmental kinetics of enzyme activities were assayed during macrocyst and fruiting-body formation. Considerable differences in the patterns of those enzyme activities were demonstrated between the two developmental modes of D. discoideum. PMID- 6286697 TI - [Haemoperitoneum due to ruptured hepatoma in a cirrhotic patient with porphyria cutanea tarda. Three years' survival (author's transl)]. AB - A case of ruptured hepatocarcinoma of the left lobe in a cirrhotic patient with a 7 years' history of porphyria cutanea tarda (PCT) is reported. Emergency left lobectomy resulted in 3 years' survival with no sign of recurrence at present. Comments are made concerning the clinical association PCT-hepatoma: (1) either PCT is associated with a hypersecretory hepatoma and behaves as a neoplastic syndrome following the same course as the tumour; (2) or PCT is associated with a cirrhosis (usually alcoholic) which facilitates the development of a hepatoma. In such cases, PCT precedes the tumour and follows a separate course. PMID- 6286698 TI - Enzymatic detection of urinary conjugated steroids after gel chromatography. AB - An enzymatic detection method is described for urinary conjugated steroids after chromatographic fractionation with Sephadex G-25. The principle of the method is as follows. Part of a 24-h urine sample, (1-2 ml of urine) is applied directly, to a short column of Sephadex G-25 and eluted with acetate buffer solution. Steroid conjugates in each fraction are hydrolyzed with steroid sulfatase--beta glucuronidase. After enzymatic hydrolysis, an enzymatic color development reagent for steroids, either 3 alpha-hydroxysteroid dehydrogenase or 3 beta hydroxysteroid oxidase, are added and the dye formed is measured spectrophotometrically. Excretion patterns of steroid-3 beta-sulfates, and steroid-3 alpha-glucuronides and steroid-3 alpha-sulfates ae shown with some patients' samples. A precision of the assay values for steroid-3 alpha glucuronide, steroid-3 alpha-sulfate and steroid-3 beta-sulfates in urine samples and assay values for normal subjects are also studied. This simple enzymatic method for detecting the excretion patterns of urinary conjugated steroids may have a diagnostic value for clinical tests. PMID- 6286699 TI - Rapid and sensitive sialidase assay by high-performance liquid chromatography and its application to detection of sialidase in human urine. PMID- 6286701 TI - A rapid bioassay to monitor murine leukemia virus infection in mice using cellular gp 71 binding. AB - A rapid and sensitive bioassay based on the availability of cell surface receptors for the binding of purified envelope glycoprotein, gp71, of Rauscher murine leukemia virus (R-MuLV) was developed to serially monitor viral-induced leukemogenesis in individual BALB/cAnN mice. The specificity of the bioassay was demonstrated by the competition of [125I]gp71 cellular binding with murine ecotropic viruses, purified unlabelled R-MuLV envelope glycoprotein and by antiserum to R-MuLV gp71. In contrast, there was no effect on the [125I]gp71 binding level with the addition of murine xenotropic viruses, R-MuLV p30, or several other proteins. The [125I]gp71 binding level of circulating leukocytes was significantly (P less than 0.05) reduced in mice after R-MuLV infection. The reduction of cellular gp71 binding developed in two stages and the latter stage was highly dependent (P less than 0.05) on circulating infectious virus titer. Using this technique, the gp71 cellular binding levels of 48-60 individual mice can be assayed in a 4 h period. The advantages of this bioassay compared to standard immunological and tissue culture techniques used in studying retrovirus expression and viral-cell interactions are discussed. PMID- 6286700 TI - Chemical inhibition of foamyvirus in primary baboon (Papio cynocephalus) kidney cells. AB - This report describes the conditions for the use of aluminum chloride (AlCl3) in growth and maintenance media for the suppression or inhibition of simian foamyviruses (SFV) in primary baboon kidney (BAK) and rabbit kidney (RK) cell cultures. When RK cells were planted in medium containing AlCl3, infected with SFV, and passaged, the growth of SFV was suppressed or inhibited by the presence of AlCl3. With this method, BAK cells yielded higher viral titers after infection with various viruses, thus making these cells more suitable for virological applications. PMID- 6286702 TI - Enzyme-linked immunosorbent assay (ELISA) for detection of herpes simplex virus specific IgM antibodies. AB - Herpes simplex virus (HSV)-specific IgM in human serum could be detected by a microplate enzyme-linked immunosorbent assay, using extracts of HSV-infected cells as antigen. Peroxidase-conjugated anti-human IgM was used to detect human IgM bound to viral antigen. Pretreatment of sera with protein A-bearing staphylococcus or with aggregated human IgG was necessary to eliminate false positive results caused by the presence of rheumatoid factor. Specificity controls included sera of patients with other herpes group virus infections. PMID- 6286703 TI - Detection and characterization of EBV antigens by micro-ELISA and chromatofocusing. AB - A micro-ELISA technique was developed for the detection of Epstein-Barr virus (EBV)-determined antigens. The enzyme-linked immunosorbent assay (ELISA) was applied with peroxidase-protein A to detect the antigens adsorbed to micro-ELISA plates. Human and rabbit antisera containing antibodies to known EBV components were used as reagents. The early antigen (EA) complex, associated with the viral cycle, was readily detected in extracts of n-butyrate- or n-butyrate + TPA induced cells. The nuclear antigen, EBNA, could be unequivocally detected only after the partial purification of the antigen by DNA cellulose chromatography. EA (and VCA) could be separated by chromatofocusing of induced cell extracts into several fractions detected by the micro-ELISA technique. This indicates that the purification of individual antigens of the EA complex can be monitored by ELISA. PMID- 6286704 TI - Decline in specificity of the ELISA due to storage of serum, and its recovery by adsorption with kaolin. AB - Several laboratories have experienced problems with the enzyme-linked immunosorbent assay (ELISA) due to non-specific background binding of human sera, but no general solution has been introduced. In this study the ELISA was used to measure antibody to herpes simplex virus in human sera which had been stored at 20 degrees C for up to 3 years, and the specificity was assessed by comparing the absorbance achieved with the virus neutralization titer. The severity of background binding was minimal in sera which had been obtained recently, but increased with storage time and gradually eliminated the specificity of the assay. When sera were adsorbed with kaolin, background binding of all sera was markedly reduced and the specificity was restored. PMID- 6286706 TI - Egg globulins in rapid virus diagnosis. AB - Egg-derived antibodies specific for a range of human viruses are now available commercially. These globulins are prepared by inoculating chickens with the relevant virus then harvesting antibodies from egg yolks. Reagents for influenza A and B, parainfluenza 1 and 3, adenovirus group antigen and rotavirus were tested. The successful use of these reagents on tissue culture and clinical material is described as well as their quality assessment. The advantages of this type of antibody are discussed. PMID- 6286705 TI - Determination of IgG- and IgM-class antibodies to mumps virus by solid-phase enzyme immunoassay. AB - An indirect enzyme immunoassay (EIA) for the determination of IgG and IgM antibodies to mumps virus is described. Viral antigens and control antigens were adsorbed onto polystyrene microtiter plates, and antibodies attached to the antigens were detected by subsequent binding of commercial peroxidase-labeled antibodies to the heavy chains of human IgG and IgM immunoglobulins. A comparison of antibody titers obtained by the EIA and by indirect immunofluorescence test showed a close concordance between these two tests, with EIA, however, being more sensitive. Occasional cross-reactions between mumps and parainfluenza antibodies were detected in the IgG antibody test but not in the IgM antibody test. In sera from 84 patients with mumps infection, all cases were diagnosed by the EIA IgM antibody assay, 96% from the first serum specimen. Mumps was diagnosed by complement fixation (CF) in 71% of these cases: unclear or erroneous results with parainfluenza titer increases in 10% and no diagnosis in 18% of the cases. The EIA IgM antibody assay was thus better than the CF test for the diagnosis of acute mumps infection. PMID- 6286707 TI - Extraction of cell-associated varicella-zoster virus DNA with triton X-100-NaCl. AB - Varicella-zoster virus (VZV) DNA was extracted from infected cells with 0.25% Triton X-100-0.2 M NaCl and purified by isopycnic centrifugation in CsCl. In each of eight experiments, 1.8-9.8 micrograms VZV DNA was obtained from 107 infected cells. The VZV DNA obtained by this procedure had a molecular weight of 88-100 x 106 as determined by sucrose gradient sedimentation and electron microscopy, and cleavage patterns after digestion with four restriction enzymes that corresponded to patterns previously described with six strains of VZV; the pattern of BamHI cleaved Triton-NaCl-extracted VZV DNA was identical to the pattern seen after DNA extraction from virions. These studies expand the usefulness of Triton X-100-NaCl for extraction of large molecular weight viral DNA from a system where considerable cell-free virus is produced (Pignatti et al., 1979, Virology 93, 260) to a system known for its marked cell association. PMID- 6286708 TI - Lectin affinity chromatography of Sindbis and Rous sarcoma virus glycopeptides and oligosaccharides. AB - Glycopeptides and endogly cosidase-digested oligosaccharides from [3H]mannose labeled Rous sarcoma virus and Sinbis virus have been fractionated by lentil lectin-Sepharose and concanavalin A-agarose affinity chromatography and subsequently analyzed by BioGel P-4 gel filtration. Only a specific subset of the Con A-bound asparaginly-oligosaccharides from he two viruses was also bound to lentil lectin, and this freaction apparently represented fucose-containing, diantennary acidic-type structures ((NeuNAc +/- Gal-GlcNAc)2 Man3 -GlcNAc2 (fucose)-ASN). The largest glycopeptides from Rous sarcoma virus were unbound to either Con A or lentil lectin and presumably contained tri- and/or tetra antennary acidic-type structures ((NeuNAc +/- Gal-GlcNAc)3--4 -Man 3GlcNAc2 (+/- fucose)-ASN). In contrast, the majority of 'hybrid'-type oligosaccharides and essentially all of the neutral oligomannosyl core structures (Man5--9 GlcNAc1 and Man3 GlcNAc1) from the endoglycosidase-digested glycopeptides of both viruses were specifically bound to Con A-agarose, with the largest neutral oligosaccharides (Man7--9GlcNAc1) bound more tightly and less efficiently eluted by alpha-methyl mannoside. PMID- 6286709 TI - Fibroblasts from a patient with leprechaunism are resistant to insulin, epidermal growth factor, and somatomedin C. AB - Leprechaunism is a rare inherited disorder characterized by severe intrauterine growth retardation and insulin resistance. Cultured skin fibroblasts from an infant with Leprechaunism were previously reported to show decreased stimulation of DNA synthesis by insulin despite apparently normal binding of [125I]insulin and [125I]somatomedin C. We have now further investigated the growth of this patient's fibroblasts and compared their metabolic responses to insulin and various peptide growth factors with responses in normal foreskin-derived fibroblasts. The doubling time of Leprechaun fibroblasts was prolonged (90 vs. 29 h), and their morphology was abnormal. Stimulation of [3H]glucose uptake was minimal with low insulin levels (1--10 ng/ml) relative to controls, but was comparable at higher insulin concentrations (1--10 micrograms/ml). Stimulation of [3H] aminoisobutyric acid uptake by insulin, epidermal growth factor (EGF), multiplication-stimulating activity, and somatomedin C (Sm-C) in Leprechaun cells was diminished relative to control cells at all concentrations tested. Furthermore, stimulation of [3H]thymidine incorporation in Leprechaum cells by EGF, Sm-C, and fibroblast growth factor was also subnormal. Binding of [125I]EGF to Leprechaun fibroblasts was not decreased. It is concluded that fibroblasts from this patient are resistant to the metabolic effects of insulin, EGF, Sm-C, and fibroblast growth factor. Since receptors for three of these peptides are apparently normal, it is likely that the defect in these cells is at the postreceptor level, perhaps involving a metabolic pathway common to the action of multiple growth factors. PMID- 6286710 TI - Induction of angiotensin I-converting enzyme by captopril in cultured human endothelial cells. AB - Captopril (2.0 microgram/ml) increased angiotensin-converting enzyme (ACE, kininase II) activity from 6- to 16-fold in culture medium of human endothelial cells from umbilical cord artery. Immunohistochemically detectable ACE was markedly increased in these cells when using rabbit antihuman lung ACE antiserum. This accords with either observations of increased ACE activity in serum and lungs from rats treated with captopril and shows induction of ACE biosynthesis in human vascular endothelial cells in culture. This observation offers a tool for studying the mechanism of ACE induction. PMID- 6286711 TI - Human chorionic gonadotropin binding to human fetal testes as a function of gestational age. AB - The characteristics of binding of hCG to testicular tissue obtained from human abortuses of 10--24 weeks gestational age were studied. Specific, saturable binding of [125I]hCG was demonstrated using homogenates of human fetal testicular tissue. The equilibrium dissociation constant ranged from 0.4 x 10(-10) M to 5.5 x 10(-10) M, a finding that is indicative of a high affinity receptor. The capacity to bind hCG was low, but varied strikingly with gestational age. The binding capacity for hCG of tissues from abortuses of gestational age less than 15 weeks and greater than 22 weeks was consistently less than 10.0 pg x mg-1 tissue (2.2 fmol x mg-1 tissue). The binding capacity for hCG of tissues from abortuses of gestational age between 15--20 weeks ranged from 2.4--29.8 pg x mg-1 tissue (0.5--6.5 fmol x mg-1 tissue) with the majority of values being greater than 10 pg x mg-1 tissue (2.2 fmol x mg-1 tissue). On the other hand, receptors for hCG in human fetal ovarian tissue were undetectable, irrespective of gestational age. It is concluded that specific high affinity binding sites for hCG are present in human fetal testes and that the binding capacity is maximum between gestational ages of 15--20 weeks. This increase in binding capacity parallels the surge in testosterone production known to occur during the same period of development. These results suggest that the increase in fetal plasma levels of testosterone during this time in gestation is the result of an increase in the sensitivity of the fetal testis to hCG caused by an increase in the number of hCG receptors, and that hCG most likely is responsible for stimulation of fetal testicular steroidogenesis in utero at this time of gestation. PMID- 6286713 TI - [The mitochondriogenesis and the turnover of cytochrome c oxidase (author's transl)]. PMID- 6286714 TI - Isopycnic separation of Escherichia coli cultures possessing colonization factor antigen I. AB - A culture of Escherichia coli possessing colonization factor antigen I was subjected to isopycnic separation on Percoll gradients. The results demonstrated successful division of the culture into two populations: (i) bacteria which cause mannose-resistant hemagglutination and (ii) bacteria which lack the ability to hemagglutinate in the presence of mannose. PMID- 6286712 TI - Isolated adrenocorticotropin deficiency associated with an empty sella. PMID- 6286715 TI - Comparison of paired whole milk and dried filter paper samples for anti enterotoxin and anti-rotavirus activities. AB - Milk specimens, 75 from cows immunized against cholera toxin and 35 from a human population in which enterotoxigenic Escherichia coli and rotaviral infections are endemic, were collected as paired filter paper and frozen whole milk samples. Each pair was tested for antibody activity against heat-labile E. coli and Vibrio cholerae enterotoxins. Additionally, 12 of the 35 paired human milk samples stored as frozen whole milk and dried on filter paper were tested for anti rotavirus immunoglobulin A. Anti-enterotoxin and anti-rotavirus immunoglobulin A titers in milk dried on filter paper compared favorably with those of their frozen whole milk pairs. Filter paper samples offered considerable advantages for field collection, transportation, and storage over frozen liquid samples. PMID- 6286716 TI - Comparative evaluation of supplemented peptone broth with sodium polyanetholesulfonate and trypticase soy broth with sodium amylosulfate for detection of septicemia. AB - We compared the yield and speed of detection of clinically important microorganisms from 10,156 paired 5-ml samples of blood cultured in supplemented peptone broth (SPB) with 0.03% sodium polyanetholesulfonate (SPS) or Trypticase soy broth (TSB) with 0.5% sodium amylosulfate (SAS). The atmosphere of incubation (open venting units) and ratio of blood to broth (1:10) were the same for both samples. Only cultures with adequate blood samples (greater than or equal to 80% of stated volume) were compared statistically. Overall, SPB/SPS outperformed TSB/SAS. Bacteroidaceae and Eubacterium were found more often (P less than 0.05) and viridans streptococci were found sooner (P less than 10(-4)) in SPB/SPS than in TSB/SAS. Most importantly, staphylococci were found both more often (P less than 0.03) and sooner (P less than 10(-7)) in SPB/SPS than in TSB/SAS. In a separate experiment, SAS slowed the growth of a clinical strain of Staphylococcus aureus in TSB. Unless important advantages can be confirmed for SAS in controlled clinical trials, SAS cannot be recommended for routine use as an anticoagulant in blood culture media. PMID- 6286717 TI - Rapid diagnosis of parainfluenza virus infection in children. AB - The indirect immunofluorescent-antibody (IFA) assay for detection of parainfluenza virus antigen was performed on samples of nasopharyngeal secretions obtained from infants and children with various respiratory illnesses to determine the usefulness of the IFA assay for rapid diagnosis of parainfluenza virus infection. Ninety-four samples of nasopharyngeal secretions were obtained from 78 children during a community outbreak of parainfluenza virus infection. Application of the IFA assay revealed the presence of parainfluenza antigen in 67 of the 94 samples. When these same specimens were inoculated into tissue culture, the presence of parainfluenza virus was confirmed in 62 (93%) of the 67 IFA positive specimens by hemadsorption on tissue culture monolayers (31 cases), presence of parainfluenza hemagglutinin in infected tissue culture fluids (23 cases), or by other methods (8 cases). Only four IFA-negative specimens were subsequently shown to be positive for parainfluenza virus by tissue culture infectivity. The IFA assay provided a more rapid and more accurate method for diagnosis of parainfluenza infection in children than did routine tissue culture methods employed currently. PMID- 6286718 TI - Shedding of rotavirus in feces of sows before and after farrowing. AB - The purpose of this study was to determine whether sows shed rotavirus near the time of farrowing. Twelve sows purchased from a common source and seropositive for rotavirus were housed in isolation in farrowing crates from 5 days before to 3 weeks after farrowing. Fecal samples were collected at 3- to 4-day intervals and examined for the presence of rotavirus by direct electron microscopy. Samples were also treated with pancreatin and inoculated onto monkey kidney cells. Rotaviral antigens were detected by a direct immunofluorescence technique, and selected positive cultures were examined by immunoelectron microscopy. Rotavirus was detected in the feces of 5 of 12 sows as early as 5 days before to 2 weeks after farrowing. Diarrhea related to rotavirus developed in 4 of 12 litters. Two of these four litters were farrowed by sows which shed rotavirus at 7 and 10 and 14 days after farrowing. The results of this study indicate that sows immune to rotavirus can shed virus in their feces at a time when piglets are particularly susceptible to infection and that adult swine are of primary importance in the epidemiology of rotavirus as initiators of infection. PMID- 6286719 TI - Identification and typing of herpes simplex viruses with monoclonal antibodies. AB - Monoclonal antibodies which reacted with type-specific antigens of herpes simplex virus type 2 or with antigens shared by herpes simplex virus types 1 and 2 were used in an indirect immunofluorescence assay to type virus isolates and to detect viral antigens in cells obtained from herpetic lesions. Complete concordance was obtained for 42 isolates typed by endonuclease restriction analysis of viral DNA and by indirect immunofluorescence with monoclonal antibodies. Examination of a limited number of ulcerative lesions revealed that indirect immunofluorescence and virus isolation were comparable in detecting herpes simplex virus. The results indicate that monoclonal antibodies can be used to accurately identify and type isolates of herpes simplex virus. PMID- 6286720 TI - Comparison of direct and indirect enzyme immunoassays with direct ultracentrifugation before electron microscopy for detection of rotaviruses. AB - A direct and an indirect enzyme immunoassay (EIA) were evaluated against a standard of electron microscopy after direct ultracentrifugation of the specimen for their performances in detecting rotaviruses. The indirect EIA had variable background activity which influenced test specificity. The indirect EIA control (test system without the detector antibody) plus a regression line (which reflected background noise) improved test specificity. However, the results of direct EIA (Rotazyme; Abbott Laboratories, North Chicago, Ill.) sensitivity (86%) and specificity (96%) were better than those of the indirect EIA in tests on 73 rotavirus-positive and 78 rotavirus-negative specimens. Endpoint titrations of purified SA-11 rotavirus showed greater sensitivity of the direct EIA test. Electron microscopy, performed after direct ultracentrifugation, and direct EIA were approximately 2 log10 more sensitive in the detection of purified SA-11 rotavirus than was electron microscopy with standard methods of unconcentrated specimen preparation. Direct EIA test are potentially sensitive, specific, and practical for the rapid detection of rotaviruses from human clinical specimens. Further studies are needed before EIA methods for detection of human rotaviruses can be equated with the level of reliability of results obtainable with sensitive electron microscopy techniques. PMID- 6286722 TI - Detection of peripheral soft tissue hemangioma with 99mTc-labeled red blood cell pool imaging. PMID- 6286721 TI - Transitions in membrane composition during postnatal development of rabbit fast muscle. AB - Early postnatal changes (4-5 days to 15 days after birth) in the biochemical composition of microsomes were investigated in rabbit skeletal muscles destined to become fast-twitch muscles. During this period, a steady decrease in the microsomal content of cholesterol and of ouabain-sensitive Na + /K + -ATPase activity, as well as a decrease in protein electrophoretic components in the 80 000-70 000 molecular weight range, were observed. These changes are probably due to a diminishing yield of microsomal membranes derived from T-tubules, as the age of the animals increases, and are indicated from a knowledge of the mixed composition of muscle microsomes and previous biochemical data on isolated T tubules. The content of cytochrome b5, which was found to be high in muscle microsomes of newborn animals, decreased strikingly as the amount of membrane bound Ca2 + -ATPase protein increased, with a crossing-over point at about 7-10 days after birth. These changes, possibly corresponding to a transition from precursor sarcoplasmic reticulum (SR) to mature SR, were found to be temporally correlated with changes in [3H] alpha-tocopherol binding ability of the microsomes and in the mitochondrial content of glycerol phosphate dehydrogenase. At the same critical periods, coincident with the onset of motile activity, the immunological cross-reactivity of the Ca2 + -ATPase protein of microsomal vesicles, with antibody specific for the Ca2 + -ATPase of adult fast SR, was found to increase markedly, as tested by competitive enzyme-linked immunosorbent assay (ELISA). The immunological data are consistent with data in the literature demonstrating an increase in the concentration of Ca2 + -ATPase molecules in the SR membranes during ontogenic development. Both these data and catalytic data, however, suggest that the Ca2 + -ATPase protein is present in the same form in the SR of immature and of adult fast muscle and, in an antigenically different form, in slow muscle SR. PMID- 6286724 TI - Genetic analysis of familial isolated growth hormone deficiency type I. AB - Nuclear DNA from individuals belonging to nine different families in which two sibs were affected with isolated growth hormone deficiency type I were studied by restriction endonuclease analysis. By using 32P-labeled human growth hormone or the homologous human chorionic somatomammotropin complementary DNA (cDNA) sequences as a probe, the growth hormone genes of affected individuals from all families yielded normal restriction patterns. Polymorphic restriction endonuclease sites (HincII and MspI), which are closely linked to the structural gene for growth hormone on chromosome 17, were used as markers in linkage analysis of DNA of family members. Of the nine affected sib pairs two were concordant, three were possibly concordant, and four were discordant for both linked markers. Since only concordant sib pairs would have inherited the same growth hormone alleles, further studies to identify mutations of the growth hormone genes should be limited to this subgroup. It is unlikely that the discordance observed in four of the sib pairs is due to recombination, because the polymorphic HincII site is only 116 base-pairs from the -26 codon of the growth hormone gene. Thus, in at least four of the nine families, the mutation responsible for isolated growth hormone deficiency is not within or near the structural gene for growth hormone on chromosome 17. PMID- 6286723 TI - Prostacyclin modulates cholesteryl ester hydrolytic activity by its effect on cyclic adenosine monophosphate in rabbit aortic smooth muscle cells. AB - We tested the hypothesis that prostacyclin (PGI2), 6-keto-prostaglandinF1 alpha(6 keto-PGF1 alpha), and several E series prostaglandins (PG) may affect the activity of cholesteryl ester (CE) hydrolase since our previous experiments indicated that smooth muscle cells (SMC) in neointima of injured rabbit aorta (a) acquire the capacity to produce PGI2 and (b) have increased lysosomal CE hydrolytic (acid cholesteryl ester hydrolase [ACEH])activity. Using cultured SMC from rabbit thoracic aorta, we demonstrated that PGI2, 6-keto-PGF1 alpha, and 6 keto-PGE1 enhanced ACEH activity fourfold. No significant effects on ACEH activity were observed with PGE1 or PGE2. Preincubation of SMC with an inhibitor of adenylate cyclase activity (dideoxyadenosine) abolished the effect of these PG on CE hydrolytic activity. Addition of dibutyryl cAMP to these SMC significantly increased ACEH activity. Although concentrations of PGI2 used significantly increased cAMP levels, proliferation of these SMC was not observed. In related experiments, we determined if the addition of PGI2, 6-keto-PGF1 alpha, or 6-keto PGE1 to cultured aortic SMC would enhance the egress of unesterified cholesterol and CE from these SMC. A significant loss of total cholesterol from PG-treated SMC was observed at the end of 14 d. Results suggest that increased synthesis of PGI2 by neointimal SMC in the injured aortic wall may, at least in part, explain the changes in CE catabolism and accumulation following injury. These PG may also be important in CE metabolism and accumulation in human arteries. PMID- 6286726 TI - Release of gelatinase from a novel secretory compartment of human neutrophils. AB - Gelatinase is a metallo-proteinase that acts specifically on denatured collagen. In human neutrophils, this enzyme is localized in small, morphologically still unidentified storage organelles that are resolved from the specific and the azurophil granules upon subcellular fractionation by differential sedimentation. When neutrophils isolated from freshly drawn blood are exposed to soluble stimuli such as N-formyl-methionyl-leucyl-phenylalanine, zymosan-activated serum, phorbol myristate acetate, or the calcium ionophore A 23187, or are induced to phagocytose opsonized zymosan, they rapidly release gelatinase in large amounts (30-70% of the cellular content in 10 min). When neutrophils from donor blood, which had been stored for 24 h at 4 degrees C are used, extensive release even occurs without added stimuli by simply warming to 37 degrees C. Gelatinase release appears to occur by secretion because it is not dependent on phagocytosis. It is paralelled by the release of specific granule contents (vitamin B(12)-binding protein), but is more rapid and much more extensive. It is, however, dissociated from the discharge of azurophil granules (as assessed by beta-glucuronidase). In addition, it was found that gelatinase release does not depend on the activation of the respiratory burst, although the two responses are often observed in parallel. Release is not due to cell damage as the cytoplasmic enzyme lactate dehydrogenase is fully retained. The distinct subcellular distribution and kinetics of release of gelatinase reported in this paper uncover a novel, truly secretory compartment of human neutrophils, which is highly responsive to stimulation. Gelatinase and possibly other enzymes stored in this secretory organelle may be involved in the early events of neutrophil mobilization, the response to chemotactic signals and diapedesis. PMID- 6286725 TI - Effects of ion substitution on bile acid-dependent and -independent bile formation by rat liver. AB - To characterize the transport mechanisms responsible for formation of canalicular bile, we have examined the effects of ion substitution on bile acid-dependent and bile acid-independent bile formation by the isolated perfused rat liver. Complete replacement of perfusate sodium with choline and lithium abolished taurocholate induced choleresis and reduced biliary taurocholate output by greater than 70%. Partial replacement of perfusate sodium (25 of 128 mM) by choline reduced bile acid-independent bile formation by 30% and replacement of the remaining sodium (103 mM) by choline reduced bile acid-independent bile formation by an additional 64%. In contrast, replacement of the remaining sodium (103 mM) by lithium reduced bile acid-independent bile formation by only an additional 20%, while complete replacement of sodium (128 mM) by lithium reduced bile formation by only 17%, and lithium replaced sodium as the predominant biliary cation. Replacement of perfusate bicarbonate by Tricine, a zwitterionic amino acid buffer, decreased bile acid-independent bile formation by greater than or equal to 50% and decreased biliary bicarbonate output by approximately 60%, regardless of the accompanying cation. In separate experiments, replacement of sodium by lithium essentially abolished Na,K-ATPase activity measured either as ouabain suppressible ATP hydrolysis in rat liver or kidney homogenates, or as ouabain suppressible 86Rb uptake by cultured rat hepatocytes. These studies indicate that bile acid(taurocholate)-dependent bile formation by rat liver exhibits a specific requirement for sodium, a finding probably attributable to the role(s) of sodium in hepatic sodium-coupled taurocholate uptake and/or in maintenance of Na,K ATPase activity. The surprising finding that bile acid-independent bile formation was substantially unaltered by complete replacement of sodium with the permeant cation lithium does not appear to be explained by Na,K-ATPase-mediated lithium transport. Although alternative interpretations exist, this observation is consistent with the hypothesis that much of basal bile acid-independent bile formation is attributable to an ion pump other than Na,K-ATPase, which directly or indirectly mediates bicarbonate transport. PMID- 6286727 TI - Association between colony forming units-granulocyte macrophage expression of Ia like (HLA-DR) antigen and control of granulocyte and macrophage production. A new role for prostaglandin E. AB - The expression of Ia-like antigens on human colony forming units-granulocyte macrophage (CFU-GM) is related to S-phase of the cell cycle, and associated with the control of normal granulocyte and macrophage production by prostaglandin E and acidic isoferritins in vitro. Ia-antigen expression by CFU-GM is lost within 3-6 h of culture at 37 degrees C and occurs simultaneously with loss of responsiveness to inhibition by these factors. Culture of bone marrow CFU-GM in a limited exposure suspension culture with 1 microM-1pM prostaglandin E (PGE1 or PGE2), but not prostaglandin F2 alpha or dibutyryl-cyclic-3'-5'-AMP results in the detection of CFU-GM Ia-antigen after 24 h. Antigen expression is associated with an absolute increase in total and S-phase CFU-GM, and restoration of responsiveness to inhibition by prostaglandin E and acidic isoferritins. The detection of Ia-antigen on CFU-GM after suspension culture with prostaglandin E results both from Ia-antigen reexpression as well as stimulation of noncycling cells to enter S-phase, express Ia-antigen and give rise to CFU-GM sensitive to inhibition by prostaglandin E and acidic isoferritins. The sensitivity of CFU-GM to inhibition by these factors after suspension culture with prostaglandin E is identical to that of the same cells tested prior to the suspension culture. These studies provide evidence for a direct regulatory association between Ia-antigen expression and control of myeloid progenitor cell differentiation, and suggest a role for prostaglandin E in the control of CFU-GM cell cycle, Ia-antigen expression, and growth regulation. PMID- 6286728 TI - Chlorination of taurine by human neutrophils. Evidence for hypochlorous acid generation. AB - The model hydrogen peroxide-myeloperoxidase-chloride system is capable of generating the powerful oxidant hypochlorous acid, which can be quantitated by trapping the generated species with the beta-amino acid, taurine. The resultant stable product, taurine chloramine, can be quantitated by its ability to oxidize the sulfhydryl compound, 5-thio-2-nitro-benzoic acid to the disulfide, 5,5' dithiobis(2-nitroben-zoic acid) or to oxidize iodide to iodine. Using this system, purified myeloperoxidase in the presence of chloride and taurine converted stoichiometric quantities of hydrogen peroxide to taurine chloramine. Chloramine generation was absolutely dependent on hydrogen peroxide, myeloperoxidase, and chloride and could be inhibited by catalase, myeloperoxidase inhibitors, or chloride-free conditions. In the presence of taurine, intact human neutrophils stimulated with either phorbol myristate acetate or opsonized zymosan particles generated a stable species capable of oxidizing 5-thio-2-nitrobenzoic acid or iodide. Resting cells did not form this species. The oxidant formed by the stimulated neutrophils was identified as taurine chloramine by both ultraviolet spectrophotometry and electrophoresis. Taurine chloramine formation by the neutrophil was dependent on the taurine concentration, time, and cell number. Neutrophil-dependent chloramine generation was inhibited by catalase, the myeloperoxidase inhibitors, azide, cyanide, or aminotriazole and by chloride-free conditions, but not by superoxide dismutase or hydroxyl radical scavengers. Thus, it appears that stimulated human neutrophils can utilize the hydrogen peroxide myeloperoxidase-chloride system to generate taurine chloramine. Based on the demonstrated ability of the myeloperoxidase system to generate free hypochlorous acid we conclude that neutrophils chlorinate taurine by producing this powerful oxidant. The biologic reactivity and cytotoxic potential of hypochlorous acid and its chloramine derivatives suggest that these oxidants play an important role in the inflammatory response and host defense. PMID- 6286729 TI - Role of angiotensin II in potassium-mediated stimulation of aldosterone secretion in the dog. AB - Potassium is known to enhance the aldosterone-stimulating action of angiotensin II. Such a synergistic interaction of potassium with angiotensin II could represent an action by angiotensin II to potentiate potassium as a stimulus. To examine for this effect of angiotensin II on potassium, plasma aldosterone levels were measured before and after an infusion of potassium chloride (15 meq i.v.) into dogs without and with prevention of angiotensin II formation by captopril, an angiotensin converting-enzyme inhibitor. In addition, responses to potassium were measured in a group of dogs receiving angiotensin II plus captopril. After potassium infusion, control dogs showed an increase of 7.7 +/- 1.9 (SEM) ng/dl (P less than 0.001) in the level of plasma aldosterone. In contrast, captopril treated dogs showed no change in plasma aldosterone concentration in response to potassium. When angiotensin II was administered to captopril-treated dogs responsiveness to potassium administration was restored (plasma aldosterone concentration increased by 7.4 +/- 2.1 ng/dl, P less than 0.002). ACTH stimulated aldosterone secretion despite captopril treatment (P less than 0.001), however, ACTH produced a greater increase in the plasma aldosterone concentration in controls than in captopril-treated animals. It is evident from these results that stimulation of aldosterone secretion by potassium is considerably enhanced by angiotensin II. There appears to exist an important interdependence of these stimuli in the regulation of aldosterone secretion. PMID- 6286730 TI - Induction by glucocorticoids of angiotensin converting enzyme production from bovine endothelial cells in culture and rat lung in vivo. AB - The effect of corticosteroids on angiotensin converting enzyme was investigated in endothelial cell cultures and intact rat lung. Cultured endothelial cells from bovine aorta showed net production of angiotensin converting enzyme (ACE) over 2 d culture in serum-free medium. Dexamethasone (DM) increased cell ACE activity six- to sevenfold at 100 nM with a threshold effect at 0.3 nM. The effect of DM on ACE production was completely inhibited by actinomycin D or cycloheximide. Deoxycorticosterone (DOC) and aldosterone were markedly less active, with a threshold near 100 nM and significant (two to threefold) stimulation of ACE activity at 1 muM. In cells incubated in the presence of 10 nM DM, DOC (10 muM) significantly inhibited ACE production compared with 10 nM DM alone, suggesting that DOC is a partial agonist/partial antagonist in this enzyme system. Protein content of cells or medium was unchanged by steroids at all doses used. In vivo, adrenalectomized rats showed lower pulmonary ACE compared with intact controls, and when injected with DM (40 mug/d for 4 d) showed a significant (twofold, P < 0.002) increase in lung ACE over oil-injected, adrenalectomized controls; serum ACE did not change. Injection with DOC (40 mug/d) or aldosterone (10 mug/d) had no effect on lung or serum ACE. Over a range (0.6 to 2,000 mug) of concentrations of DM administered daily for 7 d, the dose-response curve of DM for induction of pulmonary ACE mirrored that for thymolysis; for both, half-maximal effects were seen at approximately 6 mug DM/d, and plateau levels at 60 mug/d. We conclude that glucocorticoids are potent inducers of ACE activity in endothelial cells in culture and in rat lung in vivo, and that the action of aldosterone and DOC reflects occupancy of glucocorticoid receptors. This effect may be of (patho)physiological relevance in regulating levels of ACE in local vascular beds, and thereby modulating local levels of the vasoactive peptides angiotensin II and bradykinin. PMID- 6286731 TI - Arachnoidal cap cell hyperplasia of the intracanalicular optic nerve sheath. AB - Fifty-two intracanalicular optic nerves from 26 consecutive cadavers at the Dade County Medical Examiner's Office were examined for arachnoidal cap cells three cells thick or greater. Twenty specimens (77%) fulfilled these criteria with predominant involvement of the inferior and lateral aspects of the optic nerve sheath. Twenty-four specimens (92%) had psammoma bodies, and 17 specimens (65%) demonstrated corpora amylacea. Psammoma bodies as well as corpora amylacea were found with increasing frequency with age. However, our study did not show that age was an absolute factor toward the development of arachnoidal cap cell hyperplasia. The factors that incite foci of arachnoidal cap cell hyperplasia to greater activity leading in occasional individuals to the development of a meningioma remain to be determined. PMID- 6286732 TI - Urine cyclic nucleotide concentrations in cancer and other conditions; cyclic GMP: a potential marker for cancer treatment. AB - Cyclic guanosine 3',5' monophosphate (cyclic GMP) and cyclic adenosine 3',5' monophosphate (cyclic AMP) have been determined in random urine specimens from 95 healthy individuals, 60 patients with non-cancerous conditions, 52 patients with benign tumours, and 74 patients with malignant tumours. Concentrations of cyclic GMP have also been determined in a number of other groups, including some undergoing cancer treatment. Ninety-three per cent of cancer patients had raised urinary cyclic GMP concentrations compared to the reference range for healthy subjects. For the non-cancerous and benign groups, 33% and 42% respectively had raised concentrations. The urine cyclic AMP concentrations were similar in all groups. Urine cyclic GMP appeared to rise early in the onset of malignant growth. Successful cancer treatment was accompanied by a dramatic fall in the urine cyclic GMP concentrations, whereas if the treatment was unsuccessful the level did not change. It is concluded that urine cyclic GMP may have important applications in the monitoring of cancer treatment. PMID- 6286733 TI - Recurring digital fibroma. AB - Seven cases of recurring digital fibroma were seen over a 35-year period. All demonstrated the classical clinical, macroscopic, and microscopic features of this distinct tumour, including the pathognomonic round, eosinophilic cytoplasmic inclusion bodies. Ultrastructurally, all seven cases were confirmed to be myofibroblastic in nature, and the morphology and intracellular topography of the inclusion bodies suggested their derivation from contractile protein. These findings establish recurring digital fibroma as a neoplastic lesion of the myofibroblast. PMID- 6286734 TI - A comparative pilot study of enalapril, a new converting enzyme inhibitor, and hydrochlorothiazide in essential hypertension. AB - Eight patients with essential hypertension completed a double-blind, randomly allocated crossover comparison of either 5 or 10 mg enalapril maleate, 50 mg hydrochlorothiazide, or their combination administered once daily during sequential two-week periods. Blood pressure, pulse rate, plasma renin activity, angiotensin-converting enzyme activity, plasma aldosterone concentration, and urinary electrolytes were monitored for 24 hours after placebo and on days 1 and 14 of each treatment period. After two weeks of each treatment, only the combination of enalapril and hydrochlorothiazide significantly lowered the mean seated diastolic blood pressure (SDBP). Likewise, SDBP control (less than or equal to 90 mm Hg) was achieved only after combination therapy; six of the eight patients were controlled by the combination for up to 24 hours. The initial SDBP response to combination therapy differed with the sequence of drug addition; however, by day 14 the responses were comparable, regardless of whether hydrochlorothiazide or enalapril was first given. Mean converting enzyme activity was suppressed by enalapril in all patients, though it did not always correlate with changes in SDBP or plasma aldosterone. Mean plasma renin activity increased, but the increase was significant only on the combination. There were no serious adverse effects. PMID- 6286735 TI - Hypothesis: cyclic AMP turnover in S49 cells. AB - The fractional turnover constant (ke) of cAMP in S49 lymphoma cells stimulated by epinephrine was essentially identical to the decay constant for cAMP in these cells. This was in sharp distinction to the situation in the human diploid lung fibroblast WI-38, in which ke was much lower in hormone stimulated cells. In this study we report a new method for the determination of cAMP turnover. The method was based on the assumption that for tritium label to be incorporated into cAMP on treatment of cells with [3H]-adenine, the label must first pass through ATP. An equation relating the rate of change in cAMP specific radioactivity with cAMP and ATP specific radioactivities was thereby determined. The equation was expressed in a form that gave a linear graphical plot with the fractional turnover constant as the slope of the line. PMID- 6286737 TI - Neutralization of acid in the rumen by magnesium oxide and magnesium carbonate. AB - Two reagent and two feed grade magnesium oxides and reagent grade magnesium carbonate, sodium bicarbonate, and calcium carbonate were evaluated to ascertain their ability to neutralize acid in the rumen. Rumen fluid pH was increased in vitro, compared to the control, by antacid compounds, and their increased ranked: calcium carbonate less than feed grade magnesium oxide A less than light magnesium oxide and feed grade magnesium oxide B less than heavy magnesium oxide less than magnesium carbonate less than sodium bicarbonate. Titrations at constant pH's ranging from 3.0 to 7.5 indicated that these magnesium compounds were reactive at pH's normally in the rumen although reactivity decreased with increasing pH. Intraruminal doses of feed grade magnesium oxide A and calcium carbonate did not change rumen fluid pH for other compounds ranked: feed grade magnesium oxide B less than magnesium carbonate less than heavy magnesium oxide. Feeding of heavy magnesium oxide or magnesium carbonate increased rumen fluid pH as compared to the control diet. Feeding feed grade magnesium oxide B in three quantities to cattle resulted in proportional increased in fecal pH and fluidity of feces. Two feed grade magnesium oxides differed in their ability to neutralize acid in the rumen. PMID- 6286736 TI - Dietary sodium bicarbonate and magnesium oxide for early postpartum lactating dairy cows: effects on production, acid-base metabolism, and digestion. AB - In two experiments with cows in early lactation fed 40% corn silage and 60% concentrate diets (dry basis), we examined effects of 1.0% dietary sodium bicarbonate and .8% magnesium oxide in a 2 x 2 factorial arrangement on performance, acid-base metabolism, and digestion. In Experiment 1 with 20 Holstein and 4 Jersey cows, intake and milk production were not affected by treatment through 12 wk postpartum. In digestion trials at 3, 6, 9, and 12 wk postpartum, addition of magnesium oxide increased digestibility of dry matter 1.8% units by sodium bicarbonate whereas acid detergent fiber digestion increased 9 to 12% units with addition of either buffer. In Experiment 2, with four fistulated Holstein cows in a Latin square design, intake per unit body weight was increased .18% units by dietary sodium bicarbonate while milk production was unaffected. Dry matter and acid detergent fiber digestion were increased slightly by sodium bicarbonate plus magnesium oxide addition whereas nitrogen balance increased 23 g/day in cows fed magnesium oxide alone. Because of the low fiber content of the ration, milk fat percent increased .5 to .9% by addition of either buffer in both experiments and resulted in corresponding increased in fat corrected milk. Sodium bicarbonate and magnesium oxide prevented rapid declines in rumen pH associated with feeding in Experiment 2, which related to changes in acid detergent fiber digestion. Blood acid-base metabolites were relatively unchanged by treatment in either experiment. Urinary bicarbonate and net acid excretion rates were changed on the average by +41 and +43 meq/h by sodium bicarbonate or magnesium oxide but could not be related to changes in intake and production. PMID- 6286739 TI - Bovine leukemia virus infection and mastitis. PMID- 6286738 TI - Vitamin D3 toxicity in dairy cows. AB - Large parenteral doses of vitamin D3 (15 to 17.5 x 10(6) IU vitamin D3) were associated with prolonged hypercalcemia, hyperphosphatemia, and large increases of vitamin D3 and its metabolites in the blood plasma of nonlactating nonpregnant and pregnant Jersey cows. Calcium concentrations 1 day postpartum were higher in cows treated with vitamin D3 about 32 days prepartum (8.8 mg/100 ml) than in control cows (5.5 mg/100 ml). None of the cows treated with vitamin D3 showed signs of milk fever during the peripartal period; however, 22% of the control cows developed clinical signs of milk fever during this period. Signs of vitamin D3 toxicity were not observed in nonlactating nonpregnant cows; however, pregnant cows commonly developed severe signs of vitamin D3 toxicity and 10 of 17 cows died. There was widespread metastatic calcification in the cows that died. Because of the extreme toxicity of vitamin D3 in pregnant Jersey cows and the low margin of safety between doses of vitamin D3 that prevent milk fever and doses that induce milk fever, we concluded that vitamin D3 cannot be used practically to prevent milk fever when injected several weeks prepartum. PMID- 6286740 TI - Effect of adrenocorticotropin and cortisol on luteinizing hormone surge and estrous behavior of cows. AB - In six cows, twice daily administration of 100 IU corticotropin for 3.5 days during the follicular phase delayed the preovulatory luteinizing hormone surge and onset of behavioral estrus. Corticotropin increased progesterone and decreased estradiol and basal luteinizing hormone concentrations of blood. Following corticotropin withdrawal, a shortened period of behavioral estrus (50% of control) was accompanied by an apparent luteinizing hormone surge and ovulation. To ascertain if these effects were caused by the elevated corticosteroid concentrations induced by corticotropin, we infused intravenously four heifers with cortisol sodium succinate. During this infusion, luteinizing hormone surge and estrous behavior were inhibited; however, estradiol and basal luteinizing hormone concentrations were not affected. Furthermore, there was no luteinizing hormone surge, ovulation, or behavioral estrus after cessation of cortisol treatment. These results are consistent with a role for corticosteroids in mediating inhibitory effects on reproduction produced by corticotropin administration. PMID- 6286741 TI - Induction of lactation: comparison of injections of estradiol-17 beta and progesterone for 7 or 21 days on prolactin response to thyrotropin releasing hormone and milk yield in dairy cattle. AB - Subcutaneous injections of estradiol-17 beta and progesterone (.10 and .25 mg/kg of body weight) for 7 (group I) or 21 (II) days were used. Dexamethasone (.028 mg/kg of body weight per day) or adrenocorticotropin (200 IU per day) was injected into cows in each group on days 18 to 20 (I) or 32 to 34 (II). Additionally, 100 mug of thyrotropin releasing hormone was injected intravenously on days 1, 7, 17 (I) or 1, 7, and 31 (II). Milking was initiated on days 21 (I) or 35 (II). Overall 13 of 14 cows had mean daily yields of milk greater than 5 kg; 12 had 305-day lactations. Yields of milk in cows injected for 21 days were greater on day 1 and increased more rapidly until peak was reached at 10 wk; daily mean production throughout lactation was greater (14.3 versus 10.1 kg) than for cows injected for 7 days. Lactation curves pooled within cow within treatment differed. Concentrations of estradiol, estrone and progesterone increased during steroid injections and were 2- to 3-fold higher on day 21 in II than on day 7 (I or II), but concentrations of prolactin and total glucocorticoids in plasma did not differ during this time. The quantity of prolactin released in response to injection of thyrotropin releasing hormone was greater 10 days after steroid injections than before or during steroid injections. Preinjection concentrations of prolactin were correlated with magnitude of postinjection response to thyrotropin releasing hormone, but response was not correlated with concentrations of steroids in plasma on day of injection. PMID- 6286742 TI - Rumen volatile fatty acid production and nutrient utilization in steers fed a diet supplemented with sodium bicarbonate and monensin. AB - Effects of feeding dietary supplements of monensin and sodium bicarbonate singly or in combination on production of rumen volatile fatty acids, nitrogen balance, and rumen water kinetics were studied. Four rumen fistulated steers were fed a diet (50% concentrate mix and 50% corn silage) ad libitum in a 4 X 4 Latin square design (21-day periods) with a 2 X 2 factorial arrangement of treatments. Sodium bicarbonate increased feed intake, water intake, rumen pH, fluid dilution rate, and decreased both molar proportion and production rate of propionate in the rumen. Alteration of the ratio of acetate to propionate reflects the large decrease in propionate production relative to the small increase in acetate production. In contrast, monensin did not alter significantly rumen fluid dilution rate or ruminal pH but did decrease the molar proportion of acetate and increase that of propionate. Monensin increased production of both acetate and propionate in the rumen; however, the large increase in propionate production appears to account for more of the increase in molar proportion of propionate in the rumen. Increases in total volatile fatty acid production per kilogram of dry matter consumed with monensin supplementation appears to result from decrease in feed intake, thereby increasing ruminal retention time of dry matter and potentially the extent of digestion. Efficiency of nitrogen utilization was not altered by either sodium bicarbonate or monensin. PMID- 6286743 TI - Effects of intraruminal infusions of mineral salts on volatile fatty acid production in steers fed high-grain and high-roughage diets. AB - Four rumen fistulated Holstein steers were used to assess the effects of intraruminal salt infusions on various rumen characteristics including volatile fatty acid production rates. In the first experiment, the basal diet consisted of 75% concentrate and 25% corn silage (high-grain diet). The experimental design was a 4 X 4 Latin square in which the four treatments were: 1) intraruminal infusion of 8 liters of water (control) or 8 liters of water plus 2) 288 g sodium bicarbonate, 3) 200 g sodium chloride, or 4) 600 g sodium chloride. In the second experiment, procedures were the same except the basal diet consisted of 64% alfalfa hay and 18% each of corn silage and concentrate (high-roughage diet). When the high-grain basal diet was fed, intraruminal infusions of mineral salts increased dilution rate of rumen fluid. Total fluid flow from the rumen also was increased by salt infusions with sodium bicarbonate and the most sodium chloride from (600 g/day) exerting greatest effects for both dietary regimens. Salt infusions reduced the molar percentage of rumen propionate and increased the molar percentage of acetate when the high-grain diet was fed. These same treatments were without effect on molar percentages of rumen acids when the high roughage diet was fed. The change in the molar percentages of acetate and propionate on the high-grain ration was solely from reduction in propionate production. The lower production of propionate, from salt infusions, may have occurred because of the washout of readily fermentable materials. Dry matter disappearance from feeds placed in dacron bags and suspended in the rumen was unaffected by infusion of mineral salts. PMID- 6286744 TI - A comparative study on ultrastructure and immunological properties of human papilloma virus before and after fixation. PMID- 6286746 TI - Rationale for changes in the dietary management of diabetes. Fat, carbohydrate, and fiber. AB - Roughly three-quarters of Americans with diabetes die from atherosclerosis. Although the pathogenesis of cardiovascular disease in diabetes is not completely understood, diabetes is frequently associated with hyperlipidemia, often considered a major determinant of atherosclerosis, and with hyperglycemia, which may function as an independent risk factor. The new higher carbohydrate diets for management of diabetes facilitate reduction in the proportion of fat kilocalories. When total kilocalories are controlled, improvement in glucose tolerance also occurs in individuals with diabetes who have available endogenous or exogenous insulin. It has recently been demonstrated in subjects with diabetes that a mixture of carbohydrate and fiber and a high, rather than low, level of carbohydrate facilitate glycemic control. Inclusion of fiber-rich foods in meal plans for patients with diabetes augments established modes of therapy, which focus on weight control for Type II diabetes while synchronizing food intake and insulin for Type I diabetes. PMID- 6286745 TI - Regulation of fluid and mucous secretions in airways. PMID- 6286747 TI - A new method that investigates superoxide versus respiration endomitochondrially with a site-specific chemiluminescent probe. PMID- 6286748 TI - A new method that investigates superoxide versus respiration in vitro using bioluminescence and Sepharose-bound adenosine derivatives. AB - In vitro reactions between superoxide and phosphate derivatives of adenosine yielded quantitative amounts of stable ozonide-products. ADP-ozonide was formed in an optimized in vitro synthesis from ADP + O-(2) and affected by inhibitors and uncouplers in a similar manner to in vivo, oxidative-phosphorylation results. ADP-ozonide was further reacted with phosphoric acid to form ATP. Superoxide and ADP-ozonide may be important carrier-intermediates between respiration's electron transport chain and nodule ATP formation in vivo. PMID- 6286749 TI - Graves' IgG stimulation of continuously cultured rat thyroid cells: a sensitive and potentially useful clinical assay. AB - A continuously cultured line of normal rat thyroid (FRTL) cells can be stimulated by immunoglobulin preparations from patients with Graves' disease as measured by increases in intracellular cAMP levels. Responsiveness is concentration-dependent but is delayed in time relative to thyrotropin. Additionally, the cells respond to Graves' immunoglobulins which have no long-acting thyroid stimulator (LATS) activity and are negative when adenylate cyclase stimulation in human thyroid membrane preparations is assayed. No correlation exists between the stimulation activity and the ability of a Graves' immunoglobulin preparation to inhibit thyrotropin binding; cells are responsive even in the presence of such inhibitor activity. PMID- 6286751 TI - [Characteristics of alpha-adrenergic receptors on the rat submandibular glands during post-natal development (author's transl)]. PMID- 6286752 TI - [Flat dysplatic condyloma of the cervix: a model for the study of the early stages of viral carcinogenesis? Four cases (author's transl)]. PMID- 6286753 TI - Application of the avidin-biotin-peroxidase complex (ABC) method to the light microscopic localization of pituitary hormones. AB - The avidin-biotin-peroxidase complex (ABC) method was applied to semithin (0.5-1 micron) plastic-embedded sections of intact male rat pituitaries with the use of techniques previously developed for the peroxidase-antiperoxidase complex (PAP) method. Stains for adrenocorticotropin (ACTH), thyroid stimulating hormone (TSH), luteinizing hormone (LH), and follicle stimulating hormone (FSH) were cleaner, more reliable, and more efficient. The ABC method allowed the use of the same high dilutions of primary antisera used with the PAP method. Incubation time was cut to a third of the time used for the PAP stain. Furthermore, if the incubation time matched that used with the PAP method, (24-48 hr), the antisera could be diluted 2- to 4-fold further. This enhanced specific staining and allowed the use of dilutions similar to those used in the radioimmunoassay. In agreement with Hsu and Raine (J Histochem Cytochem 29:1349, 1981), the ABC method produced staining after only a 1-4 hr incubation in primary antibody that was diluted optimally for the PAP complex method. The stain was weak, however, and cell counts showed that it was restricted to the fraction of the specific cell population which stored the most hormone. Our tests showed that the most convenient incubation times for optimal staining were 12-16 hr. Furthermore, the ABC method appeared to stabilize greatly the reaction for FSH and thus improved its precision and reliability. PMID- 6286754 TI - The internal reticular apparatus of Camillo Golgi: a complex, heterogeneous organelle, enriched in acid, neutral, and alkaline phosphatases, and involved in glycosylation, secretion, membrane flow, lysosome formation, and intracellular digestion. AB - Its topography is one of the most characteristic features of the Golgi apparatus and the reticular nature of this organelle is evident in Golgi's first drawings, in light microscopic enzyme cytochemical preparations, and in high voltage electron micrographs of thick sections. Although individual components of the Golgi apparatus may differ in staining characteristics, morphology, contents, and enzymatic activities, they are integrated into a dynamic topographical and functional unit that is closely associated with the endoplasmic reticulum. Modulation of enzymatic activities and morphological and enzymatic heterogeneity are not surprising in an organelle that is the site of both synthetic and digestive events, including glycosylation, sulfation, formation of secretory granules and lysosomes, and the degradation of endocytized material. PMID- 6286750 TI - The mouse submaxillary gland: a model for the study of hormonally dependent growth factors. PMID- 6286756 TI - The accessory function of phagocytic cells in human T cell and B cell responses. PMID- 6286757 TI - Size of the transmembrane channels produced by complement proteins C5b-8. AB - It has been shown previously that erythrocytes can be lysed by complement proteins C5b-8, albeit at a much lower rate than C5b-9. We have now performed kinetic sieving experiments with resealed erythrocyte ghosts using sucrose (0.9 nm molecular diameter) and inulin (3.0 nm molecular diameter) as markers. We found that treatment of the ghosts with C5b-8 released sucrose, but not inulin. Addition of C9 to ghosts carrying C5b-8 dramatically increased the rate of sucrose flux and, in addition, caused release of inulin. Hence, unlike C5b-9 channels, those formed by C5b-8 measure less than 3 nm in diameter. Formation of C5b-8 channels was very slow compared with that of C5b-9 channels. Also, we found that about two-thirds of the C5b-8 ghosts did not have sucrose-releasing channels, but such channels were formed on reaction with C9. PMID- 6286755 TI - Inhibition of antibody synthesis by histamine in concanavalin A-treated mice: the possible role of glucocorticosteroids. AB - Administration of histamine (50 mg/kg) to BALB/C mice injected with concanavalin A (Con A) (100 micrograms, i.v.) 24 hr previously, results in a marked decrease in antibody synthesis to sheep red blood cells (SRBC) injected 2 hr later. This phenomenon occurs with nonimmunosuppressive doses of Con A and is strain specific. It does not take place in the response to the T-independent antigen polyvinylpyrrolidone (PVP) or if histamine is administered after the antigen. Adoptive transfer of normal syngeneic cells at the same time as antigen does not reverse this effect. Excess suppressor cell generation was excluded by co cultivation of treated spleen cells with normal cells in vitro and by determining their antibody response to SRBC 5 days later. 2-Methylhistamine, a histamine type 1 (H1) receptor agonist, mimicks the effect of histamine whereas dimaprit, a histamine type 2 (H2) receptor agonist, does not. Because histamine interaction with H1 receptors causes the release of adrenocorticotropic hormone (ACTH), we examined the effects of ACTH and corticosterone in this system and found that both could mimick the effect of histamine. These results suggest that the interaction of histamine with H1 receptors causes the release of glucocorticosteroids that may interfere with either Con A-activated T helper cell function or macrophage processing of T-dependent antigen. PMID- 6286760 TI - Immunization with one alphavirus cross-primes cellular and humoral immune responses to a second alphavirus. PMID- 6286759 TI - Arachidonic acid metabolism of the murine eosinophil. II. Involvement of the lipoxygenase pathway in the response to the lymphokine eosinophil stimulation promoter. AB - Eosinophil stimulation promoter (ESP) is a murine lymphokine that enhances the migration of eosinophils. Exogenous arachidonic acid between 0.5 and 2 micrograms/ml potentiated the activity of ESP on murine eosinophil migration, whereas such concentrations did not affect migration in the absence of ESP. Among the lipoxygenase products identified from an enriched population of murine eosinophils, leukotriene B4 (optimal activity at 100 ng/ml) and 12-HETE (optimal activity at 2 micrograms/ml) stimulated migration of these cells. Another lipoxygenase product from these cells 15-HETE inhibited ESP-induced migration; between 5 and 10 micrograms/ml 15-HETE decreased by one-half both stimulated migration and 12-HETE biosynthesis. Structurally diverse drugs at concentrations that inhibited HETE biosynthesis inhibited ESP-induced migration. The concentrations that decreased migration activity by one-half were 5 microM NDGA, 10 microM ETYA, and 150 microM BW755C. Aspirin and indomethacin at concentrations reported to inhibit prostaglandin biosynthesis did not substantially inhibit ESP activity, but concentrations of indomethacin above 20 microM caused concentration dependent inhibition of migration. The selective lipoxygenases inhibitor 134,7,10,13-eicosatetraynoic acid was more potent than ETYA in inhibition of ESP induced migration, and the selective cyclooxygenase inhibitor 6,9,12 octadecatriynoic acid did not effect inhibition. These results are consistent with the hypothesis that stimulation of eosinophils by the lymphokine ESP involves the generation of lipoxygenase products from arachidonic acid, which positively and negatively regulate the migratory activities of these cells. PMID- 6286761 TI - Adult mouse hepatocytes in primary monolayer culture express genetic resistance to mouse hepatitis virus type 3. PMID- 6286758 TI - Microinjection of macromolecules into normal murine lymphocytes by means of cell fusion. II. Enhancement and suppression of mitogenic responses by microinjection of monoclonal anti-cyclic AMP into B lymphocytes. AB - Reproducible methods are now available for introducing protein molecules such as antibodies into normal murine lymphocytes by fusion with protein molecule containing erythrocyte ghosts. Monoclonal antibodies against cyclic AMP were raised by hybridoma technique and packed into erythrocyte ghosts. Then, monoclonal anti-cyclic AMP containing ghosts were fused with splenic B lymphocytes by polyethylene glycol-mediated fusion at various intervals after LPS stimulation. This method made it possible for us to quantitatively microinject antibodies into B lymphocytes. Microinjection of anti-cyclic AMP antibody molecules into lymphocytes at a very early stage of LPS stimulation resulted in a marked enhancement of DNA synthetic responses as well as increased numbers of plaque-forming cells. Intracellular cyclic AMP levels were found to be markedly decreased after microinjection of monoclonal anti-cyclic AMP, suggesting that lowering the intracellular cyclic-AMP level in the B lymphocytes at an early stage of stimulation might have induced the enhanced proliferative as well as differentiative responses to LPS. Similar enhancing effects on cell proliferation were obtained when antibodies were injected 18 hr after stimulation. Microinjection of anti-cyclic AMP at 12 hr after culture, however, inhibited the DNA synthetic responses, and induction of plaque-forming cells was suppressed when anti-cyclic AMP was injected 6 hr after LPS stimulation. The present data suggest the biphasic regulatory roles of cyclic AMP at the early stage of B lymphocyte activation. This approach may be useful in identifying regulatory molecules in B lymphocyte induced by mitogenic or antigenic stimulation. PMID- 6286762 TI - Characterization of a progressive tumor from C3H fibroblasts transformed in vitro with SV40 virus. Immunoresistance in vivo correlates with phenotypic loss of H 2Kk. AB - Cultured SV40-transformed fibroblasts from C3H mice (SV-C3H) were "adapted" to in vivo growth by serial passage through sublethally irradiated, syngeneic recipients. After four in vivo passages, a population of cells was obtained (V4) that was weakly oncogenic in nonirradiated mice. Cells isolated from large V4 tumors (V5) were found to be highly oncogenic, producing lethal tumors at doses of less than 10(3) cells. V5 is insensitive to SV40-specific transplantation immunity in syngeneic animals but can be rejected completely by H-2 allogeneic mice. In vitro studies revealed that although V4 and the parent SV-C3H cells can induce SV40-specific cytotoxic T cells (CTL) in vitro and are lysed by these CTL, V5 does neither. The failure of V5 to interact with CTL was traced to the loss of H-2Kk antigen expression on these cells. The correlation between H-2Kk loss and immunoresistance in vivo suggests a central role for the cytotoxic T cell in in vivo tumor elimination in this system. PMID- 6286763 TI - Immunoglobulin secretion by cell lines derived from African and American undifferentiated lymphomas of Burkitt's and non-Burkitt's type. AB - As part of an investigation of the cellular origins of undifferentiated lymphomas of the Burkitt's and non-Burkitt's types, we have examined immunoglobulin secretion by cell lines and biopsy samples from these tumors and have compared it with other lymphoblastoid cell lines. The majority of American lymphoma cell lines and a smaller fraction of African lymphoma cell lines secreted monoclonal IgM. No other Ig classes were secreted. Ig secretion by the tumor samples was also found to be exclusively IgM. The secreted IgM was polymeric and associated with J chain. Secretion of free light chain by most of the tumor cell lines was observed. Quantitation by immunoassay demonstrated that the American lymphoma cell lines secreted significantly more IgM than the African lymphoma cell lines. We have proposed that the lymphoma cells represent an early stage of B cell differentiation, during which antigen-independent secretion occurs. PMID- 6286765 TI - Molecular interactions in human T cell-mediated cytotoxicity to EBV II. Monoclonal antibody OKT3 inhibits a post-killer-target recognition/adhesion step. PMID- 6286764 TI - Regulation of macrophage populations. V. Evaluation of the control of macrophage Ia expression in vitro. AB - In response to a lymphokine (LK) produced by activated T cells, macrophages can be induced to express Ia in vitro. This appears to be a complex process comprised of a number of discernible events. Peritoneal macrophages elicited by different means, and unstimulated macrophage and monocyte populations, each had a distinct kinetic profile of Ia induction. This response was characterized most noticeably by a latent period before actual Ia expression. The latent period varied from 3 to 7 days, depending on the target population, and was correlated with the state of activation of the macrophage as reflected by 5' nucleotidase activity. In spite of the protracted time course for Ia expression, all macrophage populations could be "triggered" (committed to a subsequent program of Ia expression) by exposure to the LK for as little as 2 hr. To be triggered, however, macrophages first had to go through a period of culture as adherent cells. Both the spontaneous loss and LK-dependent acquisition of Ia correlated with the functional capacity of these macrophages as antigen-presenting cells, indicating that these events are functionally significant. PMID- 6286767 TI - Antigen presentation by human B cells: T cell proliferation induced by Epstein Barr virus B lymphoblastoid cells. AB - T cell proliferation in response to antigen requires the presence of an antigen presenting accessory cell. The accessory cell most commonly associated with antigen presentation has been the macrophage (Mo). This study demonstrates that B cells in the form of Epstein Barr virus-transformed B lymphoblasts (EBV-B) are able to present tetanus toxoid (TT) to human T cells and induce proliferation of these cells. T cells freshly isolated from peripheral blood were shown to undergo blast transformation and proliferation in response to TT and irradiated EBV-B cells. Furthermore, TT-reactive T cell blasts were shown to proliferate in the presence of EBV-B cells and TT, but not with other antigens. There was a progressive increase to a plateau in T blast proliferation with increasing numbers of EBV-B cells added to the culture. The concentration of TT required for optimal antigen presentation was similar for EBV-B cells and for Mo. TT-specific cloned T cells also proliferated in response to TT and EBV-B cells and could be continuously grown in culture with TT, interleukin 2-containing supernatant, and EBV-B cells in place of autologous Mo. EBV-B cells pulsed with TT could also act as antigen-presenting cells. The proliferative response of T cell clones to TT in the presence of EBV-B cells was inhibited by antiserum to human p29,34 glycoprotein but not by anti-beta 2-microglobulin. This inhibition was shown to result from interaction with Ia-like determinants on EBV-B cells. These results indicate that B lymphoblastoid cells in man are able, like Mo, to present antigen to T cells in the context of Ia-like determinants. PMID- 6286766 TI - Distribution of terminal deoxynucleotidyl transferase and purine degradative and synthetic enzymes in subpopulations of human thymocytes. AB - Terminal deoxynucleotidyl transferase (TdT) and purine metabolic enzymes were examined in subsets of human infant thymocytes (defined by surface cell antigens) and normal peripheral T lymphocytes. Putative prothymocytes (RFB-1+, HTA-1+/- large blast-like cells), medium and high density cortical thymocytes (RFB-1+, HTA 1+), and medullary thymocytes (RFB-1-, HTA-1-, OKT3+) were isolated by density gradient centrifugation, monoclonal antibody and complement-mediated cytolysis, and cell-antibody affinity chromatography. Peripheral T lymphocytes were isolated from normal adult mononuclear cells using nylon fiber technique. Adenosine deaminase (ADA) and TdT were highest in prothymocytes 48.8 +/- 14.7 mumol/hr/10(8) cells (mean +/- SE) and 22.9 +/- 1.4 U/10(8) cells, respectively. Both enzymes decreased progressively down the maturation pathway. In peripheral T lymphocytes, ADA was 3.9 +/- 1.5 mumol/hr/10(8) cells, and TdT was undetectable. Purine nucleoside phosphorylase (PNP) and ecto-5'nucleotidase (5'NT) were lowest in cortical thymocytes (27.5 +/- 11.0 nmol/hr/10(6) cells and 2.8 +/- 1.3 nmol/hr/10(6) cells, respectively) and increased with T cell maturation. The PNP level was 124.9 +/- 17.2 nmol/hr/10(6) cells and 5'NT was 30.1 +/- 3.9 nmol/hr/10(6) cells in peripheral T lymphocytes. The deoxynucleoside kinases (deoxyguanosine, deoxyadenosine, and deoxycytidine kinases) paralleled the changes in ADA and TdT activity among the different T subsets. The proliferative activity (labeling index) was highest in the prothymocyte fraction and lowest in peripheral T cells. Variation in the distribution of these enzymes in T cell subsets may explain their different sensitivities to deoxyadenosine and deoxyguanosine toxicity and the different effects on T cell development of ADA or PNP deficiency. PMID- 6286768 TI - Mechanism of the luminol-dependent chemiluminescence of human neutrophils. PMID- 6286769 TI - Amphotericin B alters the affinity and functional activity of the oligopeptide chemotactic factor receptor on human polymorphonuclear leukocytes. AB - Leukocyte chemotaxis is initiated by the binding of chemotactic factors to specific, high-affinity receptors. Amphotericin B, a polyene antibiotic that binds to membrane cholesterol, inhibits human neutrophil (PMN) chemotaxis. We examined the effects of this drug on PMN functions mediated by the oligopeptide chemotactic factor receptor. The antibiotic irreversibly inhibited chemotaxis and depressed the binding of the radiolabeled chemoattractant, fMet-Leu-[3H]Phe, to its receptor without affecting the receptor's specificity. The drug lowered the binding affinity of the receptor by up to fivefold and slightly increased its number. Doses of amphotericin B that depressed receptor affinity and inhibited chemotaxis did not diminish lysosomal enzyme secretion or superoxide anion production. Nystatin, a less potent polyene antibiotic, also diminished chemotactic factor binding, but to a lesser degree than amphotericin B did. A chemically unrelated antifungal agent had no effect on either binding or chemotaxis. Thus, pharmacologic manipulation can alter the affinity of the chemotactic factor receptor on human PMN; this alteration is associated with a change in receptor function. The data suggest that receptor affinity regulates or at least reflects its functional state, and that the transduction mechanisms for various biologic responses mediated by the chemoattractant receptor are heterogeneous. By pharmacologic alterations of receptor affinity, one may be able to modulate specific biologic responses elicited by chemoattractant receptor ligand interactions. PMID- 6286770 TI - Specific binding of leukotriene B4 to receptors on human polymorphonuclear leukocytes. AB - Leukotriene B4 (5(S),12(R)-di-hydroxy-eicosa-6,14-cis-8,10-trans-tetraenoic acid [LTB4]) is a product of the 5-lipoxygenation of arachidonic acid, which elicits human PMN leukocyte chemotactic responses in vitro that are 50% of the maximal level at concentrations of 3 X 10(-9) M to 10(-8) M and are maximal at 2 X 10(-8) M to 10(-7) M. The specific binding of highly purified [3H]LTB4 to human PMN leukocytes was assessed both by extracting the unbound and weakly bound [3H]LTB4 with acetone at -78 degrees C and by centrifuging the PMN leukocytes through cushions of phthalate oil to separate the unbound from bound [3H]LTB4. The levels of total binding of [3H]LTB4 and of nonspecific binding of [3H]LTB4, in the presence of a 1500-fold molar excess of nonradioactive LTB4, were approximately two times higher with the phthalate oil method. Scatchard plots of the concentration dependence of the specific binding (total - nonspecific binding) of [3H]LTB4 to PMN leukocytes were linear for the acetone extraction and phthalate oil methods and revealed dissociation constants of 10.8 X 10(-9) M and 13.9 X 10( 9) M, respectively, and mean of 2.6 X 10(4) and 4.0 X 10(4) receptors per PMN leukocyte. The 5(S),12(S)-all-trans-di-HETE analog of LTB4 and 5-HETE competitively inhibited by 50% the binding of [3H]LTB4 to PMN leukocytes at respective concentrations that evoked half-maximal chemotactic responses, whereas neither N-formyl-methionyl-leucyl-phenylalanine nor chemotactic fragments of C5 inhibited the binding. Human erythrocytes exhibited no specific binding sites for [3H]LTB4. Human PMN leukocytes possess a subset of receptors for LTB4 that are distinct from those specific for peptide chemotactic factors. PMID- 6286771 TI - Actin polymerization induced by chemotactic peptide and concanavalin A in rat neutrophils. AB - Changes in the state of actin in rat neutrophils were studied after chemotactic peptide and concanavalin A stimulation by using the DNase I inhibition assay. Actin polymerization occurred within seconds after stimulation with F-Met-Leu-Phe and concanavalin A. Pretreatment of cells with cytochalasin D prevented chemotactic peptide-induced actin polymerization. The addition of F-Met-Leu-Phe to lysed cells did not produce any change in actin state. These data offer strong evidence for receptor-induced actin polymerization and support the models implicating actin microfilament formation as a crucial event in cell activation. The observations on platelets, lymphocytes, neutrophils, and islets of Langerhans from different species suggest that actin polymerization might be a universal intracellular event accompanying cell surface receptor perturbation in eukaryotic cells. PMID- 6286772 TI - The formylpeptide chemotactic receptor on rabbit peritoneal neutrophils. I. Evidence for two binding sites with different affinities. AB - F Met-Leu-[3H]Phe and f Nle-Leu-[3H]Phe binding to rabbit peritoneal neutrophils and purified membranes were measured at 4 degrees C silicone oil centrifugation assays, and the results were analyzed by the LIGAND computer program, which permits analysis of ligand binding to multiple classes of binding sites. LIGAND analysis of peptide binding to intact neutrophil indicated that both f Met-Leu [3H]Phe and f Nle-Leu-[3H]Phe detected two population of binding sites. The apparent Kd values for f Met-Leu-[3H]Phe binding were 1.6 +/- 1.0 X 10(-9) M and 2.2 +/- 0.9 X 10(-8) M, respectively, and 3.1 +/- 0.2 X 10(-9) M and 1.2 +/- 0.6 X 10(-7) M for f Nle-Leu-[3H]Phe. Furthermore, the higher affinity sites detected on whole cells comprised approximately 15 to 30% of the total sites. Two populations of binding sites were also detected on purified neutrophil plasma membranes by both radiolabeled chemotactic peptides. LIGAND analysis of peptide binding to purified membranes yielded apparent Kd values of 5.0 +/- 2.5 X 10(-10) M and 4.8 +/- 0.6 X 10(-8) M for f Met-Leu-[3H]Phe binding, and 4.7 +/- 4.2 X 10( 10) M and 3.0 +/- 1.3 X 10(-8) M for f Nle-Leu-[3H]Phe. The percentage of higher affinity sites detected by f Met-Leu-[3H]Phe and f Nle-Leu-[3H]Phe on purified membranes were 1 to 5% of the total sites detected. These data are consistent either with the existence of two independent binding sites for formylpeptides on rabbit neutrophils or receptor negative cooperativity. PMID- 6286773 TI - Specific binding of phospholipid platelet-activating factor by human platelets. AB - The binding of the phospholipid platelet-activating factor 1-O-hexadecyl-2-acetyl sn-glycero-3-phosphorylcholine (AGEPC) to washed human platelets was more than 80% complete within 2 min, which coincided with the time of initiation of platelet aggregation by AGEPC. Scatchard plot analysis of the binding of [3H]AGEPC to platelets without and with an excess of unlabeled AGEPC revealed two distinct types of binding sites. One platelet site for AGEPC exhibited a high affinity (KD = 37 +/- 13 nM, mean +/- SD), was saturable, and had a low maximal capacity of 1399 +/- 498 (mean +/- SD) molecules of AGEPC/platelet. The other platelet site demonstrated a nearly infinite binding capacity, consistent with nonreceptor uptake of AGEPC into cellular structures. The specificity of the high affinity binding site for AGEPC was assessed by comparing the capacity of several analogues of AGEPC to inhibit the binding of [3H]AGEPC to platelets and to induce platelet aggregation. An ether linkage in position 1, a short-chain fatty acid in position 2, and a choline moiety in the polar head group proved to be critical both for the binding of [3H]AGEPC to platelets and for the initiation of platelet aggregation. Exposure of platelets to AGEPC for 5 min at 37 degrees C functionally deactivated the exposed platelets to subsequent stimulation by AGEPC, as assessed by diminished aggregation, and concomitantly reduced the specific binding of [3H]AGEPC. Evaluation of the time course of the events of deactivation revealed the loss of an aggregation response to AGEPC after 90 sec at 37 degrees C, despite the retention of up to 50% of the specific binding sites for AGEPC. PMID- 6286775 TI - Generation of long-term human cytolytic cell lines with persistent natural killer activity. AB - Human cell lines maintained by in vitro stimulation with the HLA-A, B-negative, DR-positive, Epstein Barr virus-transformed, lymphoblastoid cell line Daudi in the presence of conditioned medium demonstrated significant NK activity for over 6 wk in continuous culture. These cells lyse K562 and a broad panel of lymphoblastoid cell lines but do not lyse normal peripheral blood lymphocytes or pokeweed mitogen blasts. They possess the sheep red blood cell receptor but lack other T cell markers (Lyt-3+, OKT3-). Natural killer activity correlated with the presence of a Mac 1-positive subpopulation of cells present in these long-term lines. PMID- 6286774 TI - B cell expression of endogenous retroviral envelope antigen: biochemical and immunofluorescence characterization. AB - Plasma cells of line 15I5 chickens have been shown to express antigen that is cross-reactive with endogenous retroviral envelope glycoprotein. In the present study, we isolated this antigen and showed that it is structurally homologous to envelope glycoprotein. As assayed by immunofluorescence, most, if not all, of the plasma cell-associated envelope glycoprotein is present on the cell surface although it is not bound to surface Ig or Ia antigen. Although envelope antigen was not detectable by immunofluorescence on the surface of 15I5 bursal cells or peripheral B lymphocytes, immunochemical assays established that retroviral envelope glycoprotein is present on the surface of the bursal cells but at much lower levels than on the surface of the plasma cells. Both bursal and plasma cells synthesize envelope glycoprotein. These findings indicate that an increased level of expression of endogenous retroviral envelope glycoprotein is a concomitant of the differentiation of B lymphocytes to plasma cells in 15I5 chickens. PMID- 6286776 TI - Purification of acute phase anti-hepatitis A virus (HAV) IgM and development of an IgM solid-phase radioimmunoassay for the detection of HAV. AB - A simple two-step procedure for the purification of IgM from acute phase hepatitis A infected chimpanzee serum has been developed. The procedure involves the precipitation of the euglobulin fraction with boric acid and the fractionation of the resuspended material on Sephacryl S-300 (Pharmacia Fine Chemicals, Piscataway, NJ). The purified IgM was used to develop a solid-phase radioimmunoassay (SPRIA) in which IgM was used as capture antibody and iodinated IgM was used as a probe. This assay was compared with a conventional IgG-based SPRIA. The IgM-based assay resulted in a superior response (P/N) for the detection of antigen in crude preparations; however, endpoint analyses demonstrated similar sensitivities. PMID- 6286778 TI - A simplified method for assessing PHA induced stimulation of rat peripheral blood lymphocytes. PMID- 6286777 TI - Low binding of Bolton-Hunter-labelled cholecystokinin-33 to carboxyl-terminal CCK antibodies. AB - Labelling of cholecystokinin-33 (CCK33) with [125I]hydroxyphenylpropionic acid succinimide ester (Bolton-Hunter-reagent) results in labels with excellent immunoreactivity when tested with non-carboxyl-terminal CCK-antibodies but extremely low binding to carboxyl-terminal CCK-antibodies. This finding might explain some of the difficulties experienced with production of immunoreactive CCK33 labels prepared by the Bolton-Hunter method. PMID- 6286779 TI - Evaluation of synthetic absorbable sutures in abdominal closure. PMID- 6286780 TI - [The place of conservative surgery in the treatment of nephroblastomas (author's transl)]. AB - The authors report their experience with partial nephrectomy for nephroblastoma in children. In a series of 83 cases, 10 had this operation in order to avoid chronic renal insufficiency: six for nephroblastomas which were bilateral from the onset; two for nephroblastoma in a single remaining kidney; one for nephroblastomatosis, and one operated with an erroneous diagnosis. In bilateral nephroblastomas they advise two stage operations with extemporaneous histologic control. The easiest side to operate is operated the first, and a urographic examination is done prior to the second operation. The location of the tumor, more than its size, is a determining factor for this conservative intervention. One must be prepared to do minute dissection of the pedicle. However, if there remains too little renal parenchyma, it is not worthwhile to preserve the pedicle. Contraindications for conservative surgery are: thrombosis of the renal vein, central location of the tumor, fragility of the tumor risking rupture during surgical manipulation, and spread of the tumor beyond the capsule. In 13 partial nephrectomies there was a single failure due to thrombosis of a renal vein. Ten children are alive with a follow up of nine months to eight years. None have a progressive tumor. It is certain that with chemiotherapy tumorectomies are reasonable in certain cases of nephroblastoma. PMID- 6286781 TI - [Complete retention of urine of sudden onset secondary to herpes genitalis infection (author's transl)]. AB - The authors report three cases of complete urinary retention of sudden onset during a herpes genitalis infection secondary to traumatic sexual relations. In practice, the diagnosis was made in the absence of any urological cause, the presence of local signs indicative of herpetic infection of the genital organs and lumbar puncture. Mictional disturbances disappeared in 2 to 3 weeks without leaving any sequelae. The possibility of nervous system complications in genital herpes infections has been know since 1904. By contrast, disturbances in micturition have been reported much more recently, in 1953. Complete urinary retention of sudden onset is not rare: 17 cases out of a series of 486 cases of genital infection. This may be the only clinical sign of nervous system involvement. Urinary retention develops between a few days and 3 weeks after the onset of genital herpes. Neurological examination of the lower limbs is normal. There may be transient impotence in the male and loss of the bulbo-cavernosus reflexes and impaired sensation in an S2-S3 distribution. Bladder tone is spontaneously hypoactive and increases following the injection of Urecholine. The herpes virus (HSV) is found only during the acute phase of the genital infection but almost never a few days later, when urinary retention develops. PMID- 6286782 TI - Measurement of adenosine 3',5'-monophosphate-dependent protein kinase and phosphorylase activities in in vivo conditions. AB - Microassay procedures for cAMP-dependent protein kinase and phosphorylase were developed which detected these activities in less than 25 micrograms of frozen dried epidermis from a punch biopsy of skin without homogenization. Using these procedures, the activation of cAMP-dependent protein kinase and phosphorylase by beta-adrenergic stimulation in mouse skin was studied in vivo. Cyclic AMP dependent protein kinase was stimulated by isoproterenol and inhibited by propranolol. Isoproterenol stimulation also activated phosphorylase a in mouse skin. In normal epidermis and uninvolved and involved epidermis from psoriatic patients no significant differences were found in the activities of cAMP dependent kinase and phosphorylase a. In all experiments we observed that the unstimulated activity ratios of phosphorylase a/total phosphorylase were around 20-30%; these values were much lower than those hitherto reported and show a preponderance of phosphorylase b rather than a. We suggest that in previous reports where phosphorylase a domination was found, phosphorylase b to a activation occurred during homogenization. The data also suggest that in the steady state no obvious defect in basic activities of cAMP-dependent protein kinase and phosphorylase is observed in psoriatic skin. PMID- 6286783 TI - Enhanced cell-free translation of human skin collagenase in recessive dystrophic epidermolysis bullosa. AB - The regulatory mechanisms for collagenase synthesis in recessive dystrophic epidermolysis bullosa (RDEB) have been studied using messenger RNA (mRNA) harvested from normal and RDEB skin fibroblasts to direct protein synthesis in a rabbit reticulocyte lysate translation system. Fibroblast mRNA encoded the synthesis of approximately 60,000 and approximately 55,000 dalton forms of procollagenase in cell-free translation. In contrast to biosynthesis by intact cells, there was preferential translation of the approximately 60,000 dalton specie. For quantitative comparisons with mRNA from normal cells, mRNA was harvested from fibroblasts of 2 RDEB patents whose intact cells have been documented to have increased synthesis of collagenase. Although total translational activity was equal in normal and RDEB mRNA preparations, translatable collagenase mRNA was increased 3.5- to 10-fold, suggesting that the enhanced collagenase synthesis characteristic of RDEB is due to increased concentrations or preferential translation of collagenase mRNA. PMID- 6286784 TI - Antifungal action of amphotericin B in combination with other polyene or imidazole antibiotics. AB - We compared the in vitro antifungal action of amphotericin B (AmB) used alone or in combination with a second polyene antibiotic or with miconazole or ketoconazole. When AmB was used in combination with either filipin or Etruscomycin (Farmitalia, Milan, Italy), antagonism or potentiation of the antifungal effect against Candida albicans resulted. Addition of AmB to Etruscomycin- or filipin-treated cultures resulted in antagonism. In contrast, potentiation occurred when Etruscomycin or filipin was added to cultures treated with AmB. The outcome of incubating C. albicans with combinations of AmB and either miconazole or ketoconazole depended on the duration of exposure of the cells to the drugs. Short-term incubations resulted in antagonism, whereas potentiation of antifungal effects occurred after prolonged exposure of cells to the antibiotics. In addition, supplementation of cultures with serum protein potentiated AmB induced k+ leakage at low protein concentrations and inhibited K+ leakage at high protein concentrations. PMID- 6286785 TI - Failure of 2-deoxy-D-glucose in the treatment of experimental cutaneous and genital infections due to herpes simplex virus. AB - The effectiveness of 2-deoxy-D-glucose (2-DG) was evaluated in the treatment of cutaneous infections with herpes simplex virus type 1 (HSV-1) in mice and genital infections with HSV type 2 (HSV-2) in mice and guinea pigs. Groups of mice were inoculated in the lumbosacral or orofacial area with HSV-1 and treated topically three times a day with 0.2% or 0.5% 2-DG solution beginning 3 hr after inoculation. No effect on skin lesions, mortality, or latency was observed. Mice were inoculated intravaginally with HSV-2 and treated intravaginally three times a day with 0.2%-5% 2-DG in solution or suspended in miconazole nitrate cream beginning 6 hr, 24 hr, or 48 hr after inoculation. Replication of HSV-2 in the vagina and final mortality were not affected. Guinea pigs were infected intravaginally with HSV-2 and treated both intravaginally and topically on the external genitalia four times a day with 1% or 5% 2-DG in miconazole nitrate cream. Treatment initiated just prior to development of lesions (on day 3 after inoculation) did not alter vaginal virus replication, lesion development and severity, or virus titers in lesions. PMID- 6286788 TI - Gastroenteritis due to Norwalk virus: an outbreak associated with a municipal water system. AB - An outbreak of gastroenteritis lasting for one week in August 1980 affected approximately 1,500 persons in a community in northern Georgia. Investigation included a telephone survey of the community, a survey of textile plant employees and junior high and high school students and staff, and a neighborhood door-to door survey. An association between gastrointestinal illness and consumption of drinking water was shown for community residents, students, and school staff. Attack rates (0-68%) determined in 10 neighborhoods increased significantly (P less than 0.001) with proximity to a textile plant, the site of one of two known cross-connections between an industrial water system (which contained fecal coliform bacteria) and the community water system. A fourfold rise in titer of antibody to Norwalk virus was found in 12 of 19 serum pairs from patients. Norwalk virus illness associated with drinking water from a large municipal water system has not been documented previously. Norwalk virus may be an important cause of waterborne morbidity in the United States. PMID- 6286787 TI - Detection by immune electron microscopy of the Snow Mountain agent of acute viral gastroenteritis. AB - An extensive outbreak of acute gastroenteritis of unknown etiology occurred at Snow Mountain, Colorado, in December 1976. Virus-like particles, 27 nm in diameter, were observed by electron microscopy in two of five stool specimens from individuals in the outbreak. Oral administration of a filtrate from one of the specimens induced disease in nine of 12 normal volunteers. Experimentally induced illness was similar to that observed during the outbreak. Stool specimens examined by immune electron microscopy revealed 27-nm virus-like particles in three of nine ill volunteers at the onset of illness. When the particles were used as a source of antigen, increases in levels of serum antibody were demonstrated in persons with either experimentally induced or naturally occurring illness. Therefore, it is likely that this virus-like particle is the etiologic agent of the Snow Mountain outbreak. The Snow Mountain agent appears to be morphologically similar to, but antigenically distinct from, the Norwalk and Hawaii agents by immune electron microscopy and may represent an additional antigenic type among the agents that resemble the Norwalk particle. PMID- 6286786 TI - Resistance of herpes simplex virus to adenine arabinoside and E-5-(2-bromovinyl) 2'-deoxyuridine: a physical analysis. AB - The physical map limits of DNA sequences within the DNA polymerase locus of herpes simplex virus (HSV) type 1 that define resistance mutations to adenine arabinoside (ara-A) and E-5-(2-bromovinyl)-2'-deoxyuridine (BVDU) were determined. Intertypic recombinants between HSV types 1 and 2, generated by marker rescue of temperature-sensitive mutations, had genome structures determined by restriction endonuclease analysis and were used to show that the resistance mutation for ara-Ar-1 is closely linked to the resistance mutations for phosphonoacetic acid (paar-1) and acycloguanosine (acvr-1) within a region of 2.6 kilobase pairs (kbp) in the HSV DNA polymerase locus. The resistance mutation for bvdur-1 is defined by a 2.9-kbp region that overlaps with the region defining resistance to the other three drugs but that is transferred separately. The DNA polymerase locus also contains a 2.2-kbp region that maps adjacent to and to the left of the region defining the bvdur-1 mutation which can be transferred separately and defines a region determining the HSV-1-specific sensitivity to BVDU in a manner analogous to that to acyclovir. PMID- 6286789 TI - Detection of antibody to cytomegalovirus early antigen in vaccinated, normal volunteers and renal transplant candidates. AB - Antibody to cytomegalovirus early antigen (CMV-EA) has been demonstrated in acute CMV infection. Other investigators have suggested that antibody to CMV-EA would distinguish acute CMV infection from past infection and could therefore be diagnostically useful. Antibody to CMV-EA has been demonstrated in reactivated CMV infection, in infants with congenital infection, and in pregnant women. Antibody to CMV-EA was studied in renal transplant candidates receiving Towne strain CMV vaccine, healthy adults receiving vaccine, and in asymptomatic, seropositive adults. Most healthy volunteers and renal transplant candidates receiving vaccine developed antibody to CMV-EA; it persisted in some of these adults many months after immunization. Antibody to CMV-EA was also demonstrated in asymptomatic adults with serologic evidence of a past infection. Antibody to CMV-EA is not solely an acute-phase reactant but, at least in some patients, is present in the absence of viral excretion or clinical disease. PMID- 6286791 TI - Effect of indomethacin on the in vitro lymphocyte transformation response to herpes simplex virus antigen and phytohemagglutinin after marrow transplant. PMID- 6286790 TI - Antibody response to varicella-zoster virus after natural or vaccine-induced infection. AB - The development of serum and nasopharyngeal antibody responses to varicella zoster virus (VZV) was studied in groups of children after naturally acquired varicella or after immunization with the Oka strain of live attenuated VZV vaccine administered in varying doses via respiratory inhalation or subcutaneous injection. Natural infection, subcutaneous immunization, and respiratory inhalation of large doses of VZV vaccine consistently resulted in the development of VZV-specific IgG antibody responses in serum. Although the serum IgG antibody responses persisted for at least eight to 12 months (to date) after either form of infection, the antibody activity appeared to be four- to eight-fold higher after natural infection than after immunization. Transient IgG antibody responses were observed in serum after respiratory inhalation of smaller doses of VZV vaccine. Natural infection, but not VZV vaccine, was associated with the development of serum and nasopharyngeal IgA responses to VZV in most subjects. PMID- 6286792 TI - Human immune response to herpes simplex virus. PMID- 6286793 TI - The role of lymphocytes in infections due to Epstein-Barr virus and cytomegalovirus. PMID- 6286794 TI - Effect of ion channel inhibitors on the cytopathogenicity of Entamoeba histolytica. AB - Entamoeba histolytica (axenic strain HM1-IMSS), a cytolytic enteric pathogen, kills target Chinese hamster ovary cells in two discrete steps: (1) carbohydrate specific adherence of amoebae to target cells, followed by (2) cytolysis of adherent target cells. Both steps require intact amoebic microfilament function. The effects of the fast Na+ channel blocker tetrodotoxin and the slow Na+-Ca++ channel blockers verapamil (10(-5) M) and bepridil (10(-5) M) on amoebic killing were evaluated. Verapamil and bepridil both decreased amoebic killing (P less than 0.001); tetrodotoxin had no effect. Bepridil, but not verapamil, inhibited amoebic adherence at 37 C (P less than 0.001). Both verapamil and bepridil inhibited amoebic lysis of cells after adherence occurred (P less than 0.001). Verapamil protected target cells from lysis by amoebae (P less than 0.005), whereas bepridil reduced the capacity of amoebae to kill Chinese hamster ovary cells (P less than 0.001). These findings suggest that changes in transmembrane ion flux in both the amoeba and the target cell are involved in amoebic killing of target cells. PMID- 6286796 TI - The pathogenesis of pneumonitis due to murine cytomegalovirus. AB - Virus replication and interstitial pneumonia were studied in BALB/c mice inoculated intranasally with murine cytomegalovirus (MCMV). In normal mice MCMV replicated in lung tissue but did not produce any consistent histologic abnormalities through day 21. The addition of a single dose of cyclophosphamide (0.20 mg/g) 24 hr after virus inoculation resulted in a slight increase in lung virus titers on day 10. No histologic abnormalities were noted before day 10. However, 22 or 32 virus-infected mice treated with a single dose of cyclophosphamide but only two of 26 simultaneously mock-infected, cyclophosphamide-treated mice developed severe interstitial pneumonia 10-14 days after inoculation. In contrast, animals given cyclophosphamide (0.05 mg/g) every five days after the initial dose developed a 10-fold increase in lung virus titer, but interstitial pneumonia was not observed. These data suggest that the interstitial pneumonitis seen with MCMV pulmonary infection may be immunologically mediated rather than the result of direct viral damage to the lung. PMID- 6286795 TI - Experimental infection of rhesus monkeys with Lassa virus and a closely related arenavirus, Mozambique virus. AB - As a model for the pathogenesis of Lassa fever in humans, nine rhesus monkeys were inoculated with Lassa virus. Three monkeys had had a previous asymptomatic experimental infection with Mozambique virus, a closely related arenavirus; these monkeys were protected from illness and viremia and manifested only mild pathologic lesions. The other animals developed severe disease and viremia. At necropsy, hepatocellular necrosis, interstitial pneumonia, a unique pulmonary arteritis, adrenal gland necrosis, encephalitis, and uveitis were prominent pathologic lesions which correlated with the organ titers of virus. One animal infected with Lassa virus developed prolonged viremia, a typical immune response, and sudden onset of lower limb paralysis after recovery; at necropsy chronic proliferative arteritis of the spinal cord, brain, and heart was evident. Similarities and differences in the pathologic lesions in this model and Lassa fever in humans indicate that care must be taken in interpreting the results of experiments concerning immune prophylaxis, pathogenesis, and treatment in rhesus monkeys. PMID- 6286797 TI - Genital herpes in guinea pigs: pathogenesis of the primary infection and description of recurrent disease. AB - Guinea pigs inoculated intravaginally with herpes simplex virus type 2 (HSV-2) developed a self-limiting infection characterized by vesiculo-ulcerative lesions on the external genital skin, urinary retention, and hindlimb paralysis. Infection rarely resulted in death. Virologic, histologic, and immunoperoxidase data suggested the following scheme for viral pathogenesis: initial replication in the introitus, vagina, and bladder; spread via sensory nerves to the lumbosacral dorsal root ganglia and spinal cord, and transmission via peripheral nerves to the external genital skin to produce the characteristic lesions. After recovery from primary infection, animals developed recurrent vesicular lesions, shed virus from genital sites in the absence of lesions, and harbored latent HSV 2 in dorsal root ganglia. Genital infection in the guinea pig shares many features with genital herpes in humans and provides a model to explore mechanisms of latency and reactivation and to evaluate several methods for control of recurrent disease. PMID- 6286798 TI - The effect of defective-interfering Semliki Forest virus on the histopathology of infection with virulent Semliki Forest virus in mice. AB - The majority of mice inoculated with a mixture of a lethal dose of virulent Semliki Forest virus (SFV) strain ts+ and defective-interfering (DI) SFV remained completely health, with virus infectivity levels in brain tissue reduced by 99.9%. The results of previous studies had suggested that these effects were primarily the result of the intrinsic interfering capacity of DI virus rather than of host defense responses. Because SFV strain ts+ and an avirulent strain of SFV have clearly distinguishable histopathologic effects in brain tissue, the capability of DI virus to change the virulent into the avirulent form of the disease was examined. Modulation of strain ts+ virus infection by DI virus was accompanied by a complete absence of histopathologic changes despite significant levels of infectious virus and thus differed qualitatively from infection with avirulent SFV. These results provide further evidence that the interference is not mediated through stimulation of an immune cell infiltration. PMID- 6286800 TI - Serum amyloid A levels in patients with infections due to cytomegalovirus, varicella-zoster virus, and herpes simplex virus. PMID- 6286799 TI - Treatment of experimental cryptococcal meningitis with amphotericin B, 5 fluorocytosine, and ketoconazole. AB - Ketoconazole, amphotericin B, 5-fluorocytosine, and combinations of these drugs were compared as therapy for chronic cryptococcal meningitis in steroid-treated rabbits. Two hours after treatment of rabbits with meningitis with single drugs the mean cerebrospinal fluid concentration of 5-fluorocytosine was 4.4 microgram/ml, that of amphotericin B was less than 0.3 microgram/ml, and that of ketoconazole ranged from less than 0.2 to 0.8 microgram/ml. Serial quantitative cultures indicated that amphotericin B was the best single-drug regimen. Ketoconazole provided little or no additive effect when used in combination with fluorocytosine or therapeutic doses of amphotericin for two weeks. However, the combination of ketoconazole plus amphotericin B was at least as effective as amphotericin B plus 5-fluorocytosine over a two-week treatment regimen. The addition of ketoconazole to subtherapeutic dose of amphotericin B significantly increased the killing of cryptococci in cerebrospinal fluid. PMID- 6286801 TI - Klippel-Trenaunay syndrome: a typical case. PMID- 6286802 TI - Interferon induction by 5-halo-6-phenyl pyrimidinones. AB - The interferon inducing characteristics of a new series of 6-phenyl pyrimidinol compounds are described and compared against a previously identified pyrimidine, 2-amino-5-bromo-6-methyl-4-pyrimidinol (ABMP). Interestingly, a split in ability to induce interferon but not in vivo antiviral activity was observed in the newest compounds. One representative compound, 2-amino-5-bromo-6-phenyl-4 pyrimidinol (ABPP) induced high levels of serum interferon in mice, cats and cattle in vivo and human lymphoid tissue in vitro and was consistently more active than ABMP. Another representative compound, 2-amino-5-iodo-6-phenyl-4 pyrimidinol (AIPP) was a poor interferon inducer in every system evaluated yet was as active as an in vivo antiviral agent as ABPP or ABMP. The serum interferon response induced by both ABMP and ABPP appeared to originate from an antilymphocyte serum resistant but radiosensitive cell population in the thymus and spleen. These results suggest that the antiviral activity of this group of agents is mediated by both interferon and interferon independent mechanisms. PMID- 6286803 TI - Lack of systematic correlation between the interferon mediated antiviral state and the levels of 2-5A synthetase and protein kinase in three different types of murine cells. AB - The levels of two dsRNA-dependent enzyme activities, the pppA(2'p5'A)n synthetase (2-5A synthetase) and protein kinase were investigated in control and interferon treated murine cells: L-929, K/Balb and NIH/3T3. Treatment of these cells with interferon resulted both in the establishment of the antiviral response and the development of anticellular effects. This latter was observed 3 days after treatment with interferon. The levels of 2-5A synthetase and protein kinase in control and interferon-treated cells seemed to vary from one cell type to the other. In L-929 cells, both the 2-5A synthetase and protein kinase were induced by interferon as has been shown previously. On the other hand, treatment of NIH/3T3 cells with interferon resulted in the induction of 2-5A synthetase in the absence of any enhanced protein kinase activity. This lack of protein kinase in interferon-treated NIH/3T3 cells was not due to the presence of high levels of protein phosphatases. A third type of mouse cells, K/Balb cells, contained very high levels of 2-5A synthetase in the absence of any apparent resistance to virus infection. On treatment with interferon the level of 2-5A synthetase in K/Balb cells remained constant while the protein kinase activity was enhanced by several fold. Both control and interferon-treated K/Balb cells showed similar sensitivity to the action of exogenous 2-5A thus suggesting that the 2-5A system (the 2-5A dependent nuclease and the phosphodiesterase which degrades 2-5A) was not altered on treatment with interferon. The significance of these results in relation to the mechanism of action of interferon is discussed. PMID- 6286804 TI - Inhibition of virus-induced murine diabetes by an interferon inducer. PMID- 6286805 TI - [A seroepidemiological survey of herpes simplex virus in pregnant women by microneutralization test and ELISA (author's transl)]. PMID- 6286806 TI - [Physical properties of boundary lipids (author's transl)]. PMID- 6286807 TI - [Gonadotropic regulation of estrogen secretion by rat ovarian follicles (author's transl)]. AB - The present studies were carried out to clarify the mechanisms of estrogen secretion by ovarian follicles. Rats of the Wistar-Imamichi strain at 27 days of age were treated subcutaneously with 75 IU of PMS. Ovaries were resected 24 or 48 hours after treatment and incubated for 2 hours or perifused for 5 hours using medium-199 containing ovine NIH-LH and NIH-FSH at 37 degrees C. Concentration of steroids in medium and tissue was determined by RIA. Estradiol 17-beta (E2) concentration in the perifusion effluent rose markedly following LH stimulation and rose slightly following FSH or dibutyryl cyclic-AMP stimulation. Prolactin, ACTH, PG E2 and PG F2 alpha were noneffective. LH appeared to stimulate slightly progesterone and androgen secretion by ovaries perifused. The stimulatory effects of LH on E2 secretion disappeared in ovaries from which follicular fluids were removed by punctuation with a small needle. E2 concentration in incubation medium and the total amount of E2 concentration in medium and tissue were significantly increased by treatment of LH, while the concentration in tissue was decreased by the treatment. However, LH showed no effect on E2 concentration in incubation medium and tissue of ovaries with punctured follicles. These results suggest that in the fully developed follicles LH stimulates the release of E2 stored in follicular fluids, in addition to E2 biosynthesis. E2 granules were stained by an enzyme-labeled antibody technique. They were observed only in the thecal or interstitial cells of ovaries 24 hours after PMS injection and then 48 hours after injection they also appeared in the granulosa cells either. These results suggest that in the fully-developed follicles E2 is produced in the thecal and granulosa cells and stored in follicular fluids. PMID- 6286808 TI - [The changes of human plasma ACTH, immunoreactive-beta-endorphin and cortisol during pregnancy, labor and delivery (author's transl)]. AB - Immunoreactive (IR)-beta-endorphin, ACTH and cortisol from the plasma of 88 pregnant women during gestation, and 28 women with spontaneous labor were measured. The concentrations of ACTH, IR-beta-endorphin and cortisol gradually increased during gestation. There were significant correlation between ACTH and IR-beta-endorphin (r = 0.79, p less than 0.0001), as well as between ACTH and cortisol (r = 0.54, p less than 0.0001) during gestation. Maternal ACTH and IR beta-endorphin concentrations rapidly increased during the course of labor, peaked immediately after delivery at 671.3 +/- 136.1 and 513.9 +/- 85.5 pg/ml, respectively, and then decreased two hours later. There was a highly significant correlation (r = 0.89, p less than 0.0001) between ACTH and IR-beta-endorphin during labor. The levels of cortisol were elevated during labor and showed no difference during labor, or two hours later. However, there was no significant relationship between ACTH and cortisol (r = 0.24) during labor. Chromatographic analysis was performed on the pooled plasma extracts from pregnant women at the various stage of labor. In ACTH analysis, a single peak was seen at the position of native ACTH in each case. All detectable immunoreactivities eluted in two peaks at the position of beta-LPH and beta-endorphin. However, beta-LPH to beta endorphin ratio was almost the same in each case. These results suggest that 1) ACTH and IR-beta-endorphin are secreted concomitantly, probably by the maternal pituitary gland during gestation as well as in response to the stress of labor; 2) the lack of correlation between ACTH and cortisol is due to the difference in metabolic clearance rate of these hormones during labor; 3) neither peripheral nor intrapituitary conversion of beta-LPH to beta-endorphin is seen during gestation and labor. PMID- 6286809 TI - [Effect of human chorionic gonadotropin and dibutyryl cyclic AMP on steroidogenesis in human corpora lutea (author's transl)]. AB - The effects of human chorionic gonadotropin (HCG), dibutyryl cyclic adenosin 3',5'-monophosphate (d-cAMP) on cyclic AMP (cAMP) accumulation and estradiol (E2), progesterone (P) productions in human corpora lutea were investigated by the method of short term incubation of human corpora lutea of menstrual cycle and pregnancy (6th-12th week). Thus the luteotropic effects of administered HCG on human corpora lutea were studied. (1) Elevated E2, P and cAMP concentrations in corpora lutea tissues of the menstrual cycle and comparable their levels of pregnancy were observed in the incubation with HCG. (2) In the incubation with d cAMP, the concentrations of E2, P in luteal tissue were significantly elevated in menstrual cycle and pregnancy. (3) High doses of HCG were administered to patients during luteal phase. In the incubation of their luteal tissue, E2, P and cAMP concentrations were elevated by exogenous d-cAMP, but not HCG. These results indicate that cAMP accumulation and steroidogenesis in the corpora lutea were not activated by exogenous HCG. Furthermore, the behaviour of the intra-cellular cAMP in the corpora lutea was not different between menstrual cycle and pregnancy. PMID- 6286810 TI - [The changes of human plasma ACTH and immunoreactive beta-endorphin during spontaneous or oxytocics induced/maintained labor and delivery (author's transl)]. AB - To evaluate the effects of oxytocics on ACTH and Immunoreactive (IR)-beta endorphin release, concentrations of these hormones in the plasma of 22 women during spontaneous labor, 22 women in labor who had received oxytocin and 19 women in labor who had received PG.F2 alpha at the time of vaginal delivery, were measured. Maternal ACTH and IR-beta-endorphin concentrations were elevated during spontaneous or oxytocics induced/maintained labor and peaked immediately after delivery and decreased two hours later. There were significant correlations between ACTH and IR-beta-endorphin concentrations during spontaneous labor and delivery (r = 0.921, p less than 0.0001), as well as in oxytocin or PG.F2 alpha induced/maintained labor and delivery (r = 0.833, r = 0.916, p less than 0.0001, respectively). The concentrations of ACTH and IR-beta-endorphin during PG-F2 alpha infusion were slightly higher than those seen in the other two types of labor. However, there was no significant difference in the levels of ACTH and IR beta-endorphin among the three types of labor. Simultaneous maternal and umbilical cord plasma samples were obtained in 25 cases at delivery. However, there was no significant correlation between ACTH or IR-beta-endorphin concentrations in paired samples. These results suggest that 1) IR-beta-endorphin is secreted in parallel with ACTH, into the peripheral blood by the maternal pituitary gland in response to the stress of labor during all types of labor and delivery; 2) there is no significant difference in the response of ACTH and IR beta-endorphin between spontaneous and oxytocics induced/maintained labor and delivery: 3) ACTH and IR-beta-endorphin in cord blood are not of maternal origin. PMID- 6286811 TI - [Endocrine studies in a case of arrhenoblastoma (author's transl)]. PMID- 6286812 TI - [beta-endorphin secretion and its clinical significance in a patient with Nelson's syndrome (author's transl)]. PMID- 6286813 TI - Acyclovir. PMID- 6286814 TI - Slow channel inhibitors. PMID- 6286815 TI - Induction of a chemotactic factor from human neutrophils by diverse crystals. AB - The purpose of this study was to isolate and compare the chemotactic factor generated by human neutrophils after phagocytosis of three structurally different crystals. An apparently identical chemotactic factor for neutrophils was isolated from the granular fraction of human neutrophils allowed to phagocytose MSU, CPPD, or amorphous DC. The isolated chemotactic factors migrated identically on SDS polyacrylamide gel electrophoresis, with a relative motility of 8.05, and competed with MSU-induced chemotactic factor for binding sites on the human neutrophils in an identical fashion. The data support the concept that the generation of the chemotactic factor represents a general response of the neutrophil to the phagocytosis of foreign particles. PMID- 6286816 TI - Histoplasmosis in Huntsville, Alabama. PMID- 6286817 TI - The gene encoding the common alpha subunit of the four human glycoprotein hormones. AB - The gene encoding the common alpha subunit of the four human glycoprotein hormones, chorionic gonadotropin (CG), luteinizing hormone (LH), follicle stimulating hormone (FSH), and thyroid stimulating hormone (TSH), has been cloned in a bacteriophage lambda vector. Restriction endonuclease digestion of total human DNA suggests that the common alpha subunit is coded for by a single gene. Three distinct polymorphic hybridization patterns have been observed for this gene in the human population. The cloned gene encompasses a total of 9.4 kilobases (kb) and contains three intervening sequences whose locations have been established by restriction enzyme mapping and by DNA sequencing. One of the intervening sequences is located in the 5' untranslated region, generating a leader sequence that is separated from the rest of the gene by 6.4 kb. The other two intervening sequences are 1.7 and 0.4 kb long and are located within codon number 6, and between codons 67 and 68, respectively. The location of the 5' end of the mature transcript has been established by priming placental mRNA with a restriction fragment obtained from the cloned cDNA. A transcript of similar size for the alpha subunit gene has been detected in both the pituitary, where the gene is expressed for the synthesis of LH, FSH, and TSH, and the placenta, where the gene is expressed for the synthesis of CG. When parts of the 5' untranslated nucleotide sequences of the alpha subunit and the human growth hormone genes are compared a highly homologous region is observed. These otherwise unrelated genes share the common feature that they encode a secreted pituitary polypeptide hormone. PMID- 6286818 TI - Sets of immunoglobulin V kappa genes homologous to ten cloned V kappa sequences: implications for the number of germline V kappa genes. AB - To count V kappa genes homologous to particular V kappa nucleotide sequences, thereby permitting estimates for the total V kappa repertoire, we hybridized 10 cloned V kappa cDNA sequences to restriction fragments of mouse embryo DNA (Southern blots). Particular probes labeled up to 17 fragments, of which up to 8 were strongly labeled. This indicates that the germline contains sets of related V kappa genes. Only 4 nonoverlapping gene sets of 16-22 genes each were found. Assuming that the probes used are a random sample of the V kappa pool, the pattern repetitions suggest that the germline contains a total of about 5 distinct V kappa gene sets and about 90 V kappa genes. Correlating sets of strongly labeled genes with V kappa groups having similar N-terminal sequences led to an estimate of about 300 germline V kappa genes, while extrapolation from the number of genes in the VK-21 group gave an estimate of 90-140 genes. Since the three independent estimates fell between 90 and 320 genes, the germline V kappa repertoire, at least as expressed in myelomas, probably lies in that range. The V kappa fragment patterns given by three probes did not differ detectably between BALB/c, NZB, and A/J DNA but all three patterns differed in AKR DNA, indicating that there is limited polymorphism within the mouse V kappa locus. Somatic mutation must expand the number of expressed V kappa sequences, since the 8-12 VK-21 genes estimated for BALB/c and for NZB is significantly less than the 22 known NZB VK-21 polypeptides. PMID- 6286819 TI - Construction of overproducers of the bacteriophage 434 repressor and cro proteins. PMID- 6286820 TI - The structures of the spliced mRNAs encoding polyoma virus early region proteins. AB - The polyoma virus early region mRNAs synthesized during productive infection of mouse cells have been characterized at the nucleotide level. One- and two dimensional agarose gel fractionation of nuclease S1-resistant RNA-DNA hybrids was used to establish basic structures. The two splice donors and the two splice acceptors were positioned more precisely by high resolution S1-gel mapping with terminally labeled DNA probes and polyacrylamide gels. The nucleotide sequences across the three splice joints were established by cloning and sequencing partial cDNA copies of the mRNAs. In combination with data on the polyadenylated 3'-end previously published, and the detailed analysis of the capped 5'-ends presented elsewhere, the present data complete the description of a family of differentially spliced mRNAs able to encode the known early region gene products, small-T, middle-T, and large-T proteins. PMID- 6286822 TI - Construction and characterization of SV40 recombinants with beta-globin cDNA substitutions in their early regions. AB - A cDNA segment coding for rabbit beta-globin has been inserted at different locations in the early region of simian virus 40 (SV40). The inserted sequences in these recombinants are transcribed from the SV40 early region promoter, and the primary transcripts are processed to mature mRNAs using viral intervening sequences and the early region polyadenylation site. After infection with various recombinants, beta-globin polypeptide is synthesized only when the beta-globin translation initiation codon is the first AUG in the messenger RNA sequence. When the early region transcripts contain the beta-globin cDNA sequence 3-proximal to the small t antigen coding sequence, beta-globin synthesis is not detectable. However, these recombinants produce small t antigen and abbreviated forms of large T antigen. PMID- 6286821 TI - An immunoglobulin VH pseudogene. AB - In the course of studying the members of the T15 group of VH gene segments, some of which participate in the immune response to phosphorylcholine in the mouse, we identified a VH gene segment that contains three mutations preventing its expression. The mutations are an in-frame stop codon, a 4-base insertion which causes a termination codon to be shifted into the reading frame, and a modification of the recognition elements involved in the joining of VH and D gene segments during variable region formation. This pseudogene, which is 88-96% homologous to the other members of the T15 VH gene group, is probably of relatively recent origin and will presumably be deleted from the VH gene family eventually. We suggest that pseudogenes can only arise in multigene families and that the occurrence of pseudogenes will be a relatively frequent phenomenon in these families. Because the antibody gene families are made up of multiple gene elements, undergo two types of DNA rearrangements during differentiation, and employ several different RNA splicing mechanisms for expression, there are many different ways a particular antibody gene segment may become a pseudogene. PMID- 6286823 TI - Chromosome structure and DNA sequence alterations associated with mutation of transformed genes. AB - We have constructed a series of tk+ cell lines by DNA-mediated gene transfer to correlate chromosomal behavior and DNA sequence alterations associated with reversion to the tk- phenotype. Tk- revertants were selected from each of four well-characterized transformed cell lines containing the viral tk gene and multiple human growth hormone genes (HGH). Tk- colonies were analyzed for the presence of tk and HGH sequences by blot hybridization to restriction endonuclease cleaved DNA. Revertants were further characterized by detailed karyotype analysis and hybridization in situ. Blot hybridization of forty tk- revertants indicates that over half of the revertants delete all of the transforming DNA from the recipient chromosome. In fifteen additional revertants, significant deletion has occurred, although transforming DNA is retained. The analysis of chromosomes by Giemsa banding together with hybridization in situ reveals that the deletion of transforming DNA is never associated with loss of an entire chromosome. Reversion to the tk- phenotype, therefore, seems to involve discrete deletions of transforming DNA without apparent chromosome loss. In this restricted set of mutants, it thus seems crucial to maintain the diploid chromosomal complement. PMID- 6286824 TI - Interspecific plasmid and genomic DNA sequence homologies and localization of nif genes in effective and ineffective strains of Rhizobium japonicum. AB - Three strains of Rhizobium japonicum were examined for the presence of interspecific conserved plasmid-borne DNA sequences and the location of their nif DNA sequences. Strains 61A76 and 110, which both form effective (nitrogen fixing) nodules on soybeans show very low (24%) total DNA sequence homology with each other; strain 61A76 contains one plasmid, and strain 110 contains no identifiable plasmids. Strain 61A24 which forms ineffective nodules on soybeans shows relatively high (50%) sequence homology to strain 110 and contains two plasmids. While the total sequence homology between purified plasmid DNA isolated from strains 61A76 and 61A24 appears limited, heterologous hybridizations between plasmids and total DNAs from these strains and DNA from strain 110 suggest that several sequences which are plasmid-borne in one strain can occur in the chromosome or in very large plasmid(s) in the other strains. The plasmid isolated from strain 61A24 exhibits some homology with several SmaI fragments of an octopine Ti plasmid from Agrobacterium tumifaciens, whereas the 61A76 plasmid shows no homology to this Ti plasmid. DNA sequences from five strains of Rhizobium japonicum were found to hybridize with pSA30, a plasmid carrying the Klebsiella pneumoniae nif-KDH genes, but these sequences were not located on the isolated plasmids. The organization of these genes appears to be similar in strains 110 and 61A24, but is different in strain 61A76. PMID- 6286825 TI - The nucleotide sequences of the actin genes from Saccharomyces carlsbergensis and Saccharomyces cerevisiae are identical except for their introns. AB - The actin gene from yeast Saccharomyces carlsbergensis was cloned in Escherichia coli and its complete nucleotide structure was determined. A comparison of its DNA sequence with that of the related yeast species Saccharomyces cerevisiae revealed that the coding as well as the 5'- and 3'-untranslated regions are identical. The intron, although at the same location in the two genes, differs in three positions. There is one deletion (or insertion), one transition, and one transversion. These are clustered within and around an oligo(dA) stretch of 14 (or 13) residues. Our observations identify the intron as the fastest evolving segment of this eukaryotic gene. PMID- 6286826 TI - Organization of genes and spacers within the mouse immunoglobulin VH locus. AB - The germline organization of mouse immunoglobulin VH genes has been investigated using cloned VH sequences. Hybridization studies with VH probes from plasmacytomas HPC76 (H76), S107, HOPC1 (H1), and lymphoma ABLS-8 (A8) demonstrated that the VH locus contains at least three distinct VH gene families. Comparison with other results suggests a total of about 10 such families, and of the order of 160 germline VH genes. Nucleotide sequencing revealed that the H76 family includes anti-inulin VH sequences, and the S107 family is known to encode antiphosphorylcholine sequences. The three VH gene families studied were mapped in the order H76-S107-A8/H1-CH by determining which VH genes had been deleted from several plasmacytomas by VH rearrangement events. Nine genomic clones from athe H76 family, and one each from the S107 and A8/H1 families, were characterized; collectively they span 103 kilobases (kb). Two clones from the H76 family and one from the S107 family each bore a pair of VH genes separated by approximately 14 kb, suggesting that related VH genes in these families are clustered with a typical spacing of approximately 14 kb. No other VH genes were detected within the spacers, arguing against intermingling of different families. Within the VH76 family cluster, however, two closely homologous VH genes were shown not to be adjacent. While spacer sequences were strongly conserved in the A8/H1 family, the H76 family had minimal conservation and that was restricted to regions immediately surrounding the genes. Hence conservation of spacer sequences cannot be essential for VH gene function, nor for maintenance of a VN family. Spacers in both the H76 and S108 family contained small repeat elements, some of which behaved like mobile DNA sequences. PMID- 6286827 TI - Genetic evidence for separate functional domains on the human adenovirus specified, 72 kd, DNA binding protein. AB - Seven mutations that affect various activities of the multifunctional DNA binding protein (DBP) encoded by human adenovirus have been physically mapped to different locations within DBP gene by marker rescue experiments. Two of these mutants (Ad5ts107 and Ad5ts125) contain a lesion which, under nonpermissive temperatures, decreases the capacity of the protein to bind single-strand DNA, blocks DNA replication, and prevents normal turn-off of viral early genes. In addition, the efficiency of transformation by the ts viruses compared to wild type virus is increased at the nonpermissive temperature. Both ts mutations are located in the 3' half of the DBP gene (C-terminal half of DBP). In contrast, the alterations in the five host range mutants (Ad2hr400-Ad2hr403, Ad5hr404) which overcome the block to viral late mRNA synthesis in monkey cells, but have no marked effect on DNA replication or early gene expression map in the 5' half of the DBP gene. These results suggest that the 72 kd DBP of adenovirus contains at least two functionally separable domains. PMID- 6286828 TI - Regulation in vivo of a cloned mammalian gene: cadmium induces the transcription of a mouse metallothionein gene in SV40 vectors. AB - Cadmium and other heavy metals induce metallothionein synthesis in mammalian cells. We have cloned a mouse metallothionein-I gene from a cadmium-resistant line of Ltk- cells and inserted it into viral and plasmid SV40 vectors. These recombinant molecules were introduced into cultured monkey cells and the expression of the foreign metallothionein gene was analyzed, at the RNA level, in the presence and absence of cadmium. The results show that the cloned gene retains its cadmium inducibility and that the regulation of this gene occurs primarily at the transcriptional level. PMID- 6286829 TI - Use of a lac promoter-operator fragment as a transcriptional control switch for expression of the constitutive lpp gene in Escherichia coli. AB - We constructed hybrid plasmids to allow controlled expression of the lpp gene coding for the outer membrane lipoprotein of Escherichia coli, which is otherwise expressed constitutively. This was achieved by the insertion of a DNA fragment carrying the lacUV5 promoter-operator region as a transcriptional control switch into the 5'-untranslated region of the lpp gene. When fully induced, the production of the lipoprotein, controlled under the tandem promoters of lppp lacpo-lpp, increased approximately 3-fold compared to that under lacpo-lpp control. However, it was still only one-third of the lipoprotein production under the constitutive lpp expression. One such plasmid, pKEN125, carrying lppp-lacpo lpp in pBR322 produced only a trace amount of the lipoprotein without induction in an E. coli lpp- cell. Upon the addition of isopropyl-beta-d-thiogalactoside, however, the amount of the lipoprotein reached almost 40% of the total membrane proteins. Cells carrying pKEN125 grew normally in the presence of the inducer, whereas cells carrying plasmid pKEN126 with tandem duplication of lppp-lacpo-lpp sequences in pBR322 lysed upon induction at high temperature. In cells with pKEN126 induced at high temperature, at least three new bands which were cross reactive with antilipoprotein serum in addition to the mature lipoprotein were detected by pulse-labeling cells with [35S]methionine. PMID- 6286830 TI - Molecular cloning of Rhizobium trifolii genes involved in symbiotic nitrogen fixation. AB - DNA sequences responsible for the development and maintenance of symbiotic nitrogen fixation have been identified and isolated from Rhizobium trifolii. Symbiotically-defective strains were generated by random mutagenesis with the transposon Tn5. The defective genes which give rise to the mutant phenotype have been cloned into bacterial plasmids and used as hybridization probes to isolate the corresponding wild-type genes from a clone bank of R. trifolii DNA. Symbiotic genes cloned in this manner are able to correct the lesion caused by the insertion of the transposon in their respective mutants and so restore the nitrogen fixation phenotype. The correction of the mutation is shown to occur by two distinguishable mechanisms--either by complementation or by homologous recombination. This approach provides a reliable method for isolation and mapping of bacterial DNA sequences involved in symbiotic nitrogen fixation. PMID- 6286831 TI - Transformation of mammalian cells to antibiotic resistance with a bacterial gene under control of the SV40 early region promoter. AB - A bacterial gene (neo) conferring resistance to neomycin-kanamycin antibiotics has been inserted into SV40 hybrid plasmid vectors and introduced into cultured mammalian cells by DNA transfusion. Whereas normal cells are killed by the antibiotic G418, those that acquire and express neo continue to grow in the presence of G418. In the course of the selection, neo DNA becomes associated with high molecular weight cellular DNA and is retained even when cells are grown in the absence of G418 for extended periods. Since neo provides a marker for dominant selections, cell transformation to G418 resistance is an efficient means for cotransformation of nonselected genes. PMID- 6286832 TI - The organization, structure, and in vitro transcription of Alu family RNA polymerase III transcription units in the human alpha-like globin gene cluster: precipitation of in vitro transcripts by lupus anti-La antibodies. AB - We have studied the location, structure, and in vitro transcription of repetitive DNA sequences within the human alpha-like globin gene cluster. At least eight different Alu family repeats were identified, each of which is transcribed in vitro to produce discrete RNA transcripts. The nucleotide sequence of one Alu repeat sequence, located on the 3' side of the alpha l globin gene (3'-alpha l), was determined and compared to published Alu repeat sequences. In vitro transcription of this repeat sequence generates RNA fragments of approximately 410, 260, 160, and 86 nucleotides. To determine whether these transcripts associate with specific proteins in vitro, we carried out immunoprecipitation experiments using an antiserum from systemic lupus erythematosus (SLE) patients. We find that the antiserum anti-La, which was shown to precipitate ribonucleoproteins (RNPs) containing the adenovirus VAI RNA from virus infected cells, preferentially precipitates the smallest two in vitro transcripts of the 3'-alpha l Alu repeat. These results suggest that the RNAs interact with specific factors in the in vitro transcription reaction mix to form RNP. PMID- 6286833 TI - Influence of glucocorticoids on mammary prolactin receptors in pregnant mice after ovariectomy. AB - Regulation of mammary prolactin receptors by steroid hormones was investigated in ovariectomized mid-pregnant mice. Ovariectomy increased the number of mammary prolactin receptors per cell with no effect or a slight decrease in dissociation constant (Kd). The simultaneous removal of adrenals prevented this increase in numbers. A single injection of glucocorticoid (corticosterone or cortisol) in ovariectomized-adrenalectomized mice restored the number of prolactin receptors in mammary glands to the same level as that in ovariectomized controls without changing the Kd. Aldosterone, deoxycorticosterone and oestradiol did not affect the number of mammary prolactin receptors after ovariectomy-adrenalectomy. Serum concentration of prolactin was not influenced by the hormone manipulation except with injections of oestradiol or cortisol and apparently did not correlate with the number of prolactin receptors. These results indicated that glucocorticoids are required for the increase in the number of mammary prolactin receptors induced by ovariectomy in mid-pregnant mice. PMID- 6286834 TI - A procedure for the purification of somatotrophs isolated from rat anterior pituitary glands using Percoll density gradients. AB - We have used fractionation on density gradients of Percoll to separate the cell types in the rat anterior pituitary gland and to produce a purified preparation of somatotrophs. The method differs from those described previously which used, for example, albumin or Ficoll gradients, in being more rapid and avoiding low temperatures, and therefore gives cells with improved viability. Anterior pituitary glands from male rats were dispersed with trypsin to produce 1.5 x 10 (6) -2.0 x 10 (6) cells/gland. These were fractionated on hyperbolic density gradients of Percoll. Two bands of cells containing somatotrophs were detected, one of which (band A; density 1.075-1.082 g/cm3) contained approximately 90% somatotrophs, whereas the other (band B; density 1.055-1.068 g/cm3) contained about 70% somatotrophs mixed with other cells, especially lactotrophs. Cells in band A appeared more responsive to secretagogues than those in band B; growth hormone secretion was stimulated markedly by cyclic AMP derivatives and prostaglandin E2, and inhibited by somatostatin. Such purified somatotrophs are well suited to biochemical studies on the mechanism of the control of growth hormone secretion. PMID- 6286835 TI - A dual effect of cobalt ions on the spontaneous release of transmitter at insect motor nerve terminals. AB - 1. The effect of extracellular cobalt on the frequency of miniature excitatory post-synaptic potentials (MEPSPs) was studied in cockroach leg muscle fibres that had been depolarized with 20.8 mM-K saline. 2. Cobalt ions had a dual effect on the spontaneous release of transmitter, an inhibitory action being followed by an acceleratory. A reciprocal relationship between Ca2+ and Co2+ was found for both the inhibitory and acceleratory effects. 3. The equilibrium dissociation constant for Co2+ as a competitive antagonist of spontaneous release ranged from 0.4 to 0.65 mM. It is concluded that Co2+ is much more potent than Mg2+ in suppressing spontaneous transmitter release at the insect neuromuscular junction. The antagonism by extracellular Co2+ appears to occur only at the external surface site on the terminal membrane. PMID- 6286836 TI - H+ excretion in the marine teleost Parophrys vetulus. AB - This study examined the branchial, renal and blood acid-base responses of a marine teleost to intravascular infusion of mineral acid (HCl at 1000 mu equiv/kg). In all animals the renal response was insignificant for at least 3 days following infusion. Two types of branchial and blood acid-base response were seen. Either the infused load disappeared rapidly from the extracellular into the intracellular compartment and was slowly cleared from the body (+ 16 h following infusion) or, more commonly, remained in the extracellular compartment until rapidly cleared by a pronounced increase in branchial excretion. In the latter the amount excreted exceeded that infused by about twofold on average. These responses are discussed in relation to those of freshwater teleosts. PMID- 6286837 TI - H-2L-restricted recognition of viral antigens In the H-2d haplotype, anti vesicular stomatitis virus cytotoxic T cells are restricted solely by H-2L. AB - H-2d-encoded gene products were analyzed as restriction antigens for anti vesicular stomatitis virus (VSV) cytotoxic T lymphocytes (CTL). Cold target competition experiments revealed that VSV recognition was H-2D region-restricted; H-2K-end-restricted recognition of VSV could not be demonstrated. That VSV is not recognized in the context of K-region-encoded gene products is also supported by the observation that H-2dm1 and H-2dm2 mice, strains that contain H-2Kd but have an alteration in H-2L and/or H-2D/L, are nonresponders in the CTL assay. Two different lines of evidence eliminated H-2Dd, H-2Md, and H-2Rd as the restriction antigens: (a) H-2dm2-VSV inhibitors that express H-2Dd and H-2Md did not block the lysis of P815-VSV targets by Balb/c anti-VSV killer cells, and (b) a hybridoma specific for H-2Dd failed to inhibit killer cell activity in this same effector/target combination. However, two monoclonal antibodies specific for H 2Ld but not H-2Rd completely blocked anti-VSV cytotoxic activity. Taken together, in the H-2d haplotype, anti-VSV CTL recognize VSV solely in the context of the H 2Ld molecule. This is the first demonstration of the exclusive use by a mouse stain of the H-2L molecule only for H-2-restricted recognition, and thus supports the notion that H-2L plays a major role in restricting antigen specific recognition. Finally, the fact that an anti-H-2Ld monoclone completely blocked an H-2dm2 anti-BALB/c CTL response indicates that H-2R, a molecule absent in H-2dm2 anti-BALB/c CTL response indicates that H-2R, a molecule absent in H-2dm2 but not BALB/c, does not sensitize H-2 alloreactive CTL. PMID- 6286838 TI - BALB- and Harvey-murine sarcoma virus transformation of a novel lymphoid progenitor cell. AB - BALB- and Harvey-murine sarcoma viruses (MSV) comprise a family of retroviruses whose mouse- and rat-derived onc genes are closely related. These viruses induce sarcomas and erythroleukemias in susceptible animals. An in vitro colony assay that detects transformation of lymphoid cells by Abelson-murine leukemia virus was used to demonstrate that BALB- and Harvey-MSV transform a novel hematopoietic cell both in culture and in vivo. Bone marrow colony formation was sarcoma virus dependent, followed single-hit kinetics, and required the presence of mercaptoethanol in the agar medium. BALB- and Harvey-MSV-induced colonies could be established in culture as continuous cell lines that demonstrated unrestricted self-renewal capacity and leukemogenicity in vivo. The cells had a blast cell morphology and lacked detectable markers of mature cells within the myeloid or erythroid series. They also lacked detectable immunoglobulin mu chain or Thy-1 antigen, markers normally associated with committed cells of the B and T lymphoid lineages, respectively. However, the transformants contained very high levels of terminal deoxynucleotidyl transferase (TdT), an enzyme believed to be specific to early stages within the lymphoid differentiation pathway. This phenotype distinguishes these BALB- and Harvey-MSV transformants from any previously reported hematopoietic targets of transforming retroviruses, including the pre-B lymphoid cell transformed by Abelson-MuLV under identical assay conditions. These newly identified lymphoid progenitor cell transformants may provide an important means of studying early stages of lymphoid ontogeny and the possible role of TdT in lymphoid development. PMID- 6286839 TI - Use of Epstein-Barr virus-transformed B cell lines for the generation of immunoglobulin-producing human B cell hybridomas. AB - HGPRTase-deficient EBV-transformed B cell lines were shown to be effective fusion partners with mitogen-activated human B cells for the construction of Ig producing human B cell hybridomas. In a series of experiments using these lines and B cells from several tissue sources, approximatley 20% of the cultures plated were consistently positive for growth after hypoxanthine-aminopterin-thymidine selection and approximatley 30% of these synthesized significant new Ig. A marked increase in Ig secretion was observed after hybridization, which was due to new Ig; Ig from the parental lime was shown to disappear in several instances. Special analyses were carried out on a human hybridoma secreting antibody specific for tetanus toxoid and tetanus toxin and stable subclones were derived. These studies suggest that EBV-transformed lines will prove useful in human hybridization studies, thus making a large library of B cell lines available for the generation of human monoclonal antibodies. PMID- 6286840 TI - Structural and biosynthetic studies on the two molecular forms of the (Na+ + K+) activated adenosine triphosphatase large subunit in Artemia salina Nauplii. AB - The large subunit of (Na+ + K+)-activated ATPase from brine shrimp, Artemia salina, migrates as two bands in sodium dodecyl sulfate-polyacrylamide gels. The slower migrating band, as observed in neutral or alkaline gel systems, is designated alpha 1 and the faster, alpha 2. Structural and biosynthetic studies have been performed to determine if these two bands represent independent molecular forms or precursor products. Peptide mapping of partial proteolytic digests of alpha 1 and alpha 2 showed no distinguishable difference between them whereas this technique produced very distinct differences in the large subunit derived from three different species. The two large subunit bands also behaved identically when cross linked with cupric phenanthroline either in the presence or absence of digitonin, whereas other proteins in these preparations were unaffected. The peptide mapping and cross-linking experiments demonstrate that alpha 1 and alpha 2 have identical or nearly identical primary and probably higher order structure. Their different mobilities may be due to post translational modification leading, for example, to different oligosaccharide composition. During development of the brine shrimp nauplius, alpha 1 increases in relative abundance while alpha 2 decreases. NaH14CO3 incorporation and pulse chase experiments indicate that alpha 1 and alpha 2, as well as the small subunit of the brine shrimp (Na+ + K+)-activated ATPase, are synthesized at the same time during development and that all changes in the rates of synthesis of these subunits occur at the same time. The apparent rates of degradation of the subunits are also similar. These results are inconsistent with a precursor product relationship between alpha 1 and alpha 2. PMID- 6286841 TI - Changes in mouse oocyte membrane potential and permeability during meiotic maturation. AB - Mouse oocytes spontaneously undergo meiotic maturation when removed from the ovary and cultured in vitro. This paper describes the changes in cell surface membrane potential from the germinal vesicle (GV) stage (- 46 mV, inside negative), through germinal vesicle breakdown (GVBD) (- 31 mV), to the polar body (PB) stage (- 17 mV). Radioisotope tracer flux experiments established the total exchangeable internal K at the GV stage in 190 mM, the K efflux rate coefficient as 6.7 X 10(-5) sec-1) and the K permeability as 46.7 X 10(-8) cm sec-1. In comparison with previous results with PB stage oocytes, this suggests that the membrane potential depolarization is caused largely by a decrease in K permeability. When GVBD is blocked with 0.11 mM dibutyryl cyclic AMP, the membrane potential does not depolarize. This suggests a linkage between depolarization and GVBD. However, incubation in 80 mM K medium, which depolarizes the oocyte, does not trigger GVBD. It is concluded that membrane depolarization by itself is not sufficient to cause GVBD. PMID- 6286842 TI - Block of outward current in cardiac Purkinje fibers by injection of quaternary ammonium ions. AB - We have studied the effects of iontophoretic injection of the quaternary ammonium compounds tetraethylammonium (TEA) and tetrabutylammonium (TBA) in cardiac purkinje fibers. We find that TBA(+) is a more effective blocker than TEA(+), but injection of either compound reduces the time-dependent outward plateau currents, transient outward current (I(to)), and the delayed rectifier (I(x)). Our findings provide evidence that these outward cardiac currents are carried by channels that in some respects are pharmacologically similar to squid axon potassium channels. We demonstrate that this procedure is a new tool that can be useful in the analysis of membrane currents in the heart. PMID- 6286843 TI - Influx of calcium, strontium, and barium in presynaptic nerve endings. AB - Depolarization-induced (potassium-stimulated) influx of 45Ca, 85Sr, and 133Ba was measured in synaptosomes prepared from rat brain. There are two phases of divalent cation entry, "fast" and "slow;" each phase is mediated by channels with distinctive characteristics. The fast channels inactivate (within 1 s) and are blocked by low concentrations (less than 1 micro M) of La. The slow channels do not inactivate (within 10 s), and are blocked by high concentrations (greater than 50 micro M) of La. Divalent cation influx through both channels saturates with increasing concentrations of permeant divalent cation; in addition, each permeant divalent cation species competitively blocks the influx of other permeant species. These results are consistent with the presence of "binding sites" for divalent cations in the fast and slow channels. The Ca:Sr:Ba permeability ratio, determined by measuring the influx of all three species in triple-label experiments, was 6:3:2 for the fast channel and 6:3:1 for the slow channel. A simple model for ion selectivity, based on the presence of a binding site in the channel, could account well for slow and, to some extent, for fast, channel selectivity data. PMID- 6286844 TI - Kinetics of the sodium pump in red cells of different temperature sensitivity. AB - Ouabain-sensitive K influx into ground squirrel and guinea pig red cells was measured at 5 and 37 degrees C as a function of external K and internal Na. In both species the external K affinity increases on cooling, being three- and fivefold higher in guinea pig and ground squirrel, respectively, at 5 than at 37 degrees C. Internal Na affinity also increased on cooling, by about the same extent. The effect of internal Na on ouabain-sensitive K influx in guinea pig cells fits a cubic Michaelis-Menten-type equation, but in ground squirrel cells this was true only at high [Na]i. There was still significant ouabain-sensitive K influx at low [Na]i. Ouabain-binding experiments indicated around 800 sites/cell for guinea pig and Columbian ground squirrel erythrocytes, and 280 sites/cell for thirteen-lined ground squirrel cells. There was no significant difference in ouabain bound per cell at 37 and 5 degrees C. Calculated turnover numbers for Columbian and thirteen-lined ground squirrel and guinea pig red cell sodium pumps at 37 degrees C were about equal, being 77-100 and 100-129 s-1, respectively. At 5 degrees C red cells from ground squirrels performed significantly better, the turnover numbers being 1.0-2.3 s-1 compared with 0.42-0.47 s-1 for erythrocytes of guinea pig. The results do not accord with a hypothesis that cold-sensitive Na pumps are blocked in one predominant form. PMID- 6286845 TI - Slowing of sodium channel opening kinetics in squid axon by extracellular zinc. AB - The interaction of Zn ion on Na channels was studied in squid giant axons. At a concentration of 30 mM Zn2+ slows opening kinetics of Na channels with almost no alteration of closing kinetics. The effects of Zn2+ can be expressed as a "shift" of the gating parameters along the voltage axis, i.e., the amount of additional depolarization required to overcome the Zn2+ effect. In these terms the mean shifts caused by 30 mM Zn2+ were +29.5 mV for Na channel opening (on) kinetics (t1/2 on), +2 mV for closing (off) kinetics (tau off), and +8.4 mV for the gNa-V curve. Zn2+ does not change the shape of the instantaneous I-V curve for inward current, but reduces it in amplitude by a factor of or approximately 0.67. Outward current is unaffected. Effects of Zn2+ on gating current (measured in the absence of TTX) closely parallel its actions on gNa. On gating current kinetics are shifted by +27.5 mV, off kinetics by +6 mV, and the Q-V distribution by +6.5 mV. Kinetic modeling shows that Zn2+ slows the forward rate constants in activation without affecting backward rate constants. More than one of the several steps in activation must be affected. The results are not compatible with the usual simple theory of uniform fixed surface charge. They suggest instead that Zn2+ is attracted by a negatively charged element of the gating apparatus that is present at the outer membrane surface at rest, and migrates inward on activation. PMID- 6286846 TI - Divalent cations and the activation kinetics of potassium channels in squid giant axons. AB - The effects of external Zn+2 and other divalent cations on K channels in squid giant axons were studied. At low concentration (2 mM) Zn+2 slows opening kinetics without affecting closing kinetics. Higher concentrations (5-40 mM) progressively slow opening and speed channel closing to a lesser degree. In terms of "shifts," opening kinetics are strongly shifted to the right on the voltage axis, and off kinetics much less so. The shift of the conductance-voltage relation along the axis is intermediate. Zinc's kinetic effects show little sign of saturation at the highest concentration attainable. Zn does not alter the shape of the instantaneous current-voltage relation of open channels. Some other divalent cations have effects similar to Zn+2, Hg2+ being the most potent and Ca+2 the least. After treatment with Hg+2, which is irreversible, Zn+2 still slows opening kinetics, which suggests that each channel has at least two sites for divalent cation action. The results are not compatible with a simple theory of fixed, uniform surface charges. They suggest that external cations interact directly with a negatively charged element of the gating apparatus that moves inward from the membrane's outer surface during activation. Examination of normal kinetics shows that there is a slow step somewhere in the chain leading to channel opening. But the slowest step must not be the last one. PMID- 6286847 TI - Sodium and potassium transport in herpes simplex virus-infected cells. AB - Sodium (Na+) and potassium (K+) flux in African green monkey kidney cells (Vero) was examined following infection by herpes simplex virus type 1 (HSV-1). A decline in the rate of K+ uptake at 5 h post-infection was shown using 86Rb+ as a K+ tracer. In contrast, host protein synthesis was inhibited by 3 h post infection. The decrease in rate of K+ transport to levels 70 to 90% of that of mock-infected cells did not, however, reflect an inability of HSV-1-infected cells to maintain normal intracellular concentrations of Na+ and K+. At 7 h post infection, intracellular Na+ and K+ concentrations were determined to be 26.6 +/- 9.4 mM- and 33.3 +/- 10.3 mM-Na+ and 130.1 +/- 4.7 mM- and 137.1 +/- 3.2 mM-K+ in mock-infected and HSV-1-infected cells respectively. Intracellular Na+ did not increase above control levels over at least a 9 h period following HSV-1 infection. The Michaelis constant (Km) of K+ transport in HSV-1-infected or mock infected Vero cells at 6 h post-infection was determined to be the same with calculated values of 1.38 +/- 0.51 mM and 1.79 +/- 0.42 mM respectively. A virus induced alteration of intracellular Na+ and K+ concentrations cannot, therefore, account for the HSV-1 induced inhibition of host protein synthesis at 3 h post infection as has been suggested in other virus systems. PMID- 6286848 TI - Transfection with the isolated herpes simplex virus thymidine kinase genes. II. Evidence for amplification of viral and adjacent cellular DNA sequences. AB - We have previously described several series of biochemical transformation experiments in which small defined portions of herpes simplex virus (HSV-1 and HSV-2) DNA encompassing the thymidine kinase (TK) gene were introduced into Ltk- cells by the calcium transfection procedure. The presence of authentic virus TK enzyme in several subcloned cell lines derived from these experiments was confirmed by either the specific incorporation of [125I]iododeoxycytidine into their nuclei or the inhibition of cell growth by the antiviral drug arabinosyl thymine. A panel of 24 independent Ltk+ cell lines receiving either isolated virus DNA fragments or cleaved plasmid DNAs was examined by blot hybridization for both the presence and copy number of virus TK DNA sequences. Most cell lines contained a single virus DNA fragment covalently joined to host (or carrier Ltk-) mouse DNA sequences, but several contained multiple copies of the TK gene. Examination of the structural arrangement of the virus DNA in two early passage multicopy cell lines indicated that the TK gene had integrated into Ltk- cell DNA and then subsequently both viral and flanking cellular sequences were amplified to create up to 20 tandem duplications. In one case, mapping of the adjacent cellular sequences has revealed that the total repeat unit is greater that 23 kilobases (kb) in size. On subsequent passaging, even in HAT medium, the amplified repeat units were not stable and fell to only three to four copies per haploid cell genome. These cell lines should prove useful for additional studies to examine the expression of co-selected non-TK virus sequences and the influence of adjacent cellular DNA sequences on transcription and retransfection of the resident TK gene. PMID- 6286849 TI - Pathogenesis of herpes simplex virus in congenitally athymic mice: the relative roles of cell-mediated and humoral immunity. AB - Athymic nude (nu/nu) mice were inoculated in the ear pinna with 10(4) p.f.u. herpes simplex virus type 1 (strain SC 16). Initially, the virus was observed to replicate in the pinna, spreading via a neurological route to the dorsal root ganglia, spinal cord, brain and adrenal glands. Following the transfer of lymphoid cells from day 7 herpesvirus-infected hairy immunocompetent donors into infected nude mice, virus was not isolated from the pinna and nervous system of the majority of the mice. The passive transfer of neutralizing polyclonal anti herpesvirus serum or neutralizing monoclonal anti-gp D serum did not reduce infectivity in the pinna, but markedly reduced the amount of virus in the ganglia and spinal cord. These data suggest that neutralizing antibodies play an important role in restricting the movement of virus to the nervous system, whereas cell-mediated immune (CMI) mechanisms are essential for eliminating virus from the pinna. PMID- 6286851 TI - Long-term mouse cytomegalovirus infection of tracheal organ culture: relation to host cell replication. AB - Tracheal organ cultures infected with mouse cytomegalovirus (MCMV) were shown to produce virus for up to 180 days with virus replication primarily in epithelial cells. Persistent virus infections were established in tracheal organ cultures from both MCMV-susceptible and MCMV-resistant strains of mice. In acutely infected tracheal organ cultures, cellular DNA synthesis appeared to precede the production of significant amounts of virus antigens and the release of virus into culture fluids. Since persistently infected tracheal organ cultures continued to synthesize cellular DNA, the results suggest that host cell turnover may continually renew the population of MCMV-susceptible cells. The results suggest a possible mechanism for virus persistence in tracheal organ culture based on continuing serial infection of newly susceptible cells combined with prolonged release of virus infected cells. PMID- 6286850 TI - Ultrastructural localization of cytomegalovirus DNA synthesis in infected guinea pig cells. AB - Using electron microscopic autoradiographic techniques, the uptake of [methyl-3H] thymidine into uninfected and guinea-pig cytomegalovirus (GPCMV)-infected guinea pig embryo (GPE) fibroblast cells was investigated. In GPCMV-infected GPE cells, [methyl-3H]thymidine uptake into cellular DNA was inhibited prior to the onset of virus DNA synthesis as well as during the entire period of virus replication. Virus DNA synthesis increased slowly during the period of active virus replication, which usually lasted from 18 to 48 h post-infection. Only the electron-dense amorphous matrices and fibrillar structures in the nuclear inclusions were associated with the tritiated labels and thus contained virus DNA. PMID- 6286852 TI - Integrated shope virus DNA is present and transcribed in the transplantable rabbit tumour Vx-7. AB - To facilitate a molecular analysis of Shope papilloma virus-induced neoplastic cells, we have established a cell line from Vx-7, a transplantable tumour originally induced by the Shope virus. Single phase molecular hybridization and Southern transfer methods were employed to assess copy number and physical state of the DNA, and the extent of transcription. Both tumour and cell line were found to contain multiple copies of the virus genome and these were all integrated into the host cell DNA. Transcripts corresponding to a complexity of approx. 1% of the virus genome were detected at low abundance. These results are discussed relative to our earlier findings with tumours induced directly by virus, and to requirements for maintenance of the Vx-7 tumour over the 30 years that it has been in existence. PMID- 6286853 TI - Cloning of DNA fragments from the left end of the adenovirus type 12 genome: transformation by cloned early region 1. AB - The human adenovirus serotype 12 (Ad-12) EcoRI-C DNA fragment (0 to 16.5 map units) was cloned in the plasmid vector pAT153. This cloned Ad-12 EcoRI-C DNA fragment was subcloned generating recombinant plasmids which contained the Ad-12 SalI-C fragment (0 to 10.3 map units), the Ad-12 HindIII-G fragment (0 to 6 X 8 map units) and the Ad-12 AccI-H fragment (0 to 4 X 7 map units). Thus, we constructed recombinant plasmids which contain Ad-12 DNA sequences which represent all or part of the virus transforming gene region. The capacity of the cloned Ad-12 EcoRI-C DNA fragment to transform rat cells in vitro was assessed using the focus assay on primary cultures of rat cells. The specific transforming activity of this recombinant plasmid was in the same range as that found for intact Ad-12 DNA. Transformed foci which were induced by the cloned Ad-12 EcoRI-C DNA fragment were established as cell lines and the presence of Ad-12 DNA in these lines was demonstrated using the Southern blotting technique. PMID- 6286854 TI - Translation of capped virus mRNA in encephalomyocarditis virus-infected cells. AB - The pattern of protein synthesis in HeLa cells simultaneously infected with encephalomyocarditis virus (EMCV) and Semliki Forest virus (SFV) has been analysed throughout the time course of the infection. The ratio of the picornavirus protein gamma versus the togavirus late protein C increased when the m.o.i. of EMCV was raised, and the ratio of C/gamma increased with higher multiplicities of SFV. Under some conditions, the co-infected cells simultaneously synthesized the picornavirus and togavirus proteins, and the cells exclusively translated the capped 26S mRNA from SFV at the end of the co infection experiment. The influence of the time of addition of the second virus on the relative translation of the capped and uncapped mRNAs was also studied. When HeLa cells were co-infected with 5 p.f.u./cell of EMCV and 200 p.f.u./cell of SFV, only the synthesis of SFV proteins was apparent, whereas if SFV was added 1 to 3 h later during the course of EMCV infection the cells synthesized picornavirus and togavirus proteins. If the cells were superinfected with SFV 1 h after EMCV addition, host protein synthesis was drastically inhibited after 3 h of EMCV infection. By 5 h post-infection both kinds of virus proteins were synthesized and at 7 h post-infection the cells preferentially translated the capped 26S mRNA from SFV. If cells were first infected with SFV (10 p.f.u./cell) and co-infected or superinfected at 1 h with EMCV (50 p.f.u./cell), shut-off of host protein synthesis occurred 3 h after infection and the cells synthesized both kinds of virus proteins. However, 9 h after infection the cells synthesized SFV proteins only. When double-infected HeLa cells were placed in a hypotonic medium, they mainly synthesized the togavirus late proteins, whereas under hypertonic conditions, they translated the picornavirus RNA exclusively. These results suggest that the two kinds of mRNAs (SFV 26S mRNA and EMCV 35S mRNA) are present in the infected cells and that the relative translation of each of them depends on the external ionic conditions. PMID- 6286855 TI - Ultraviolet-irradiated vesicular stomatitis virus and defective-interfering particles are similar non-specific inhibitors of virus infection. AB - The way in which ultraviolet-irradiated vesicular stomatitis virus (VSV) inhibits the early events in VSV infection has been further characterized. Comparison of several different u.v.-irradiated thermolabile, temperature-sensitive mutants before and after heat inactivation established a requirement for inhibitory activity of functional G, N and L proteins, but not M protein. Defective interfering (DI) particles, whether irradiated or not, inhibited VSV primary transcription as efficiently as UV-VSV, suggesting that virus proteins rather than transcription products are responsible for inhibition. Addition of inhibitory UV-VSV at different times after infection established that inhibition results from an action at an intracellular site, rather than at the cell surface or in the process of internalization. A similar inhibition by UV-VSV of infection by Sendai virus, Semliki Forest virus, Sindbis virus and influenza virus suggests that UV-VSV is acting by inducing a general change in the intracellular environment. PMID- 6286856 TI - The internal organization of the varicella-zoster virus genome. AB - DNA was extracted from varicella-zoster (VZ) virions prepared in sucrose gradients. Thirty-eight molecules examined by electron microscopy were found to have a mean length of 46.7 micrometers. Examination of self-annealed VZV DNA molecules revealed that the virus genome was composed of a unique linear large sequence with a mol. wt. of 74.4 X 10(6) to 78.4 X 10(6), and a unique short sequence of mol. wt. approx. 9.8 X 10(6) flanked by inverted repeat sequences of 4.7 X 10(6) mol. wt. PMID- 6286859 TI - Selection of temperature-sensitive mutants in persistent infection by parainfluenza virus type 3. AB - Vero cells persistently infected with parainfluenza virus type 3 (para 3) were examined for the production of temperature-sensitive (ts) mutants. After 6 months and 33 passages, ts mutants formed the great majority of the virus being shed into the supernatant fluid. Complementation studies gave evidence that the mutants shared a common lesion. Although ts mutants may play a role in the maintenance of persistent infection in this system, the mutants proved to be unstable when removed from the milieu of persistent infection. It is thus possible that their selection may be secondary to (an) unidentified factor(s) which play(s) a more primary role in the maintenance process. PMID- 6286857 TI - Characterization of an antigen associated with the Marek's disease lymphoblastoid cell line MSB-1. AB - A Marek's disease lymphoblastoid cell line (MSB-1) has been analysed by immunoprecipitation for expression of tumour-associated antigen, Marek's disease virus (MDV)-specific antigens and antigens specific to avian leukosis-sarcoma viruses. Rabbit antisera raised against two independently derived cell lines after extensive absorption with normal chick cells reacted with a polypeptide of mol. wt. 40 000 (40K) in extracts of MSB-1 cells. The 40K polypeptide was not present in myeloblasts or in chick embryo fibroblasts (CEF) infected with MDV and did not react with antiserum raised against normal chicken thymus antigens. The possibility that the 40K polypeptide is a tumour-associated antigen is discussed. Seven MDV-specific antigens were noted in infected CEF (mol. wt. 110K, 100K, 80K, 70K, 50K, 35K and 32K) but none of these was detected in MSB-1 cells. The avian leukosis-sarcoma group-specific antigen P27gag and its precursor Pr76gag were not found in MSB-1 cells, confirming that expression of mature gag protein is not required for transformation by MDV. However, two polypeptides of unknown origin and function (mol. wt. 180K and 110K) were precipitated from MSB-1 cells with a rabbit anti-Rous sarcoma (Schmidt-Rupin, subgroup D) antiserum. PMID- 6286858 TI - Immune stimulation of sensitized chicken lymphocytes by avian retrovirus proteins. AB - Peripheral blood lymphocytes of chickens bearing tumours induced by avian sarcoma virus can be specifically stimulated to divide by the crude culture fluids of virus-infected cells. In this communication, we show that relevant antigenic activity apparently resides in each of the internal virus proteins p15 and p27. The ability of infectious culture fluids to be mitogenic for sensitized lymphocytes is greatly reduced following treatment with antibodies specific for either total avian myeloblastosis virus (AMV) protein or for p27. PMID- 6286860 TI - The electron microscopical and physical characteristics of small round human fecal viruses: an interim scheme for classification. AB - Many of the small round human fecal viruses implicated in outbreaks of nonbacterial gastroenteritis have been collected together and examined under the electron microscope. Negatively stained preparations without the addition of antibody were used so that the surface morphology of the virus particles remained clearly visible. It was apparent that several viruses, previously thought to be simply antigenic variants within the Norwalk group of viruses, show distinct morphological differences and quite clearly belong to other virus groups. By comparing the features of all the viruses examined in this study, both with each other and with standard cell culture strains of enterovirus, parvovirus, and calicivirus, it has been possible to propose an interim classification scheme, based primarily on the morphological appearance of the particles and supported by estimations of size and buoyant density. PMID- 6286861 TI - A one-year virological survey of acute intussusception in childhood. AB - A prospective study was carried out during 1 year in order to correlate intussusceptions in childhood with a viral infection. A virus was incriminated in 50% of the 64 patients in this survey. Adenovirus infections remain predominant as compared to rotavirus infections. PMID- 6286862 TI - The virus susceptibility of skin-derived fibroblasts from families with insulin dependent diabetes mellitus. AB - We have studied the growth of eight different viruses on skin fibroblasts from three families each having one or more diabetic members and appropriate controls. The haplotypes of all of the family members had been previously characterized. In addition, we have investigated the growth of mumps virus on the lymphoblast cultures from four families of the same type. Our results show no difference between the growth of these viruses in cells derived from juvenile diabetics and cells derived from nondiabetic siblings and parents even when the haplotypes were identical. However, we noted a striking resistance of human skin fibroblast cultures from both normal and diabetic individuals to Coxsackie B virus infection. PMID- 6286863 TI - Production of hepatitis B e antibody in Epstein-Barr virus-induced B lymphocyte cell lines. AB - B lymphocytes separated from anti-HBe-positive donors were established as lymphoblastoid cell lines by infection with EBV, but anti-HBe in the culture supernatant from such lymphoblastoid cell lines could not be detected. The lymphoblastoid cell lines were rosetted with HBe antigen-coupled SRBC to prepare cells for the production of specific anti-HBe. Antibody activity in the culture supernatant against rosette forming cells was detected by RIA, ID, and R-PHI tests during the first 1 to 4 wk, but not after 5 wk. The activity in the supernatant was not destroyed by treatment with 2-mercaptoethanol, indicating that the antibody might be IgG. PMID- 6286864 TI - A rapid enzyme immunofiltration technique using monoclonal antibodies to serotype herpes simplex virus. AB - A rapid enzyme immunofiltration technique using monoclonal antibodies to serotype herpes simplex virus is described. It requires only a single tube culture showing viral cytopathology and can accommodate multiple specimens in a single assay. The monoclonal antibodies confer absolute specificity and the use of horseradish peroxidase-conjugated staphylococcal protein A or antiglobulin permits easy visual interpretation of the results following the 2-3 hour assay. Although it is possible that an occasional wild strain of HSV might escape recognition by monoclonal antibody, this potential problem has not been observed among the more than 500 clinical isolates tested to date, all of which have yielded an unequivocal result. PMID- 6286865 TI - Enhancement of human rotavirus infectivity in a monkey kidney cell line by human expressed breast milk. AB - Eight of forty (20%) human expressed breast milks enhanced the infectivity of human rotavirus (HRV) when assayed by immunofluorescence in a monkey kidney cell line (LLC-MK2). This enhancement was demonstrated with two HRV strains, one derived from a fecal specimen of a child with acute gastroenteritis, the other a tissue culture adapted strain. The phenomenon could have important implications in the pathogenesis of HRV infections and in future vaccine programs. PMID- 6286866 TI - Dissociation of hepatitis A virus antigen-anti-HAV antibody complexes by 2 mercaptoethanol and dithiothreitol. AB - Intravenous inoculation of two marmosets and one chimpanzee with hepatitis A virus (HAV) resulted in the replication of virus in liver, excretion of HAV particles in stool, and the appearance of circulating antibodies specific for hepatitis A. The development of an early antibody response in the chimpanzee and in one of the two infected marmosets was shown to interfere with the serologic detection of HAV antigen (HAV Ag) in homogenates of acute phase liver tissue obtained from these animals. Treatment of HAV Ag-positive and IgM anti-HAV positive liver homogenates with thiol reducing compounds was shown to release HAV Ag from in vitro formed immune complexes. The increased RIA response for HAV Ag in homogenates treated with 2-mercaptoethanol (2-ME) or dithiothreitol (DTT) was further shown not to be due to activation of HAV Ag itself or to a nonspecific effect on the RIA coating antibody, radiolabeled probe, or homogenized liver tissue. IgG and IgM double-antibody sandwich RIAs for HAV Ag were also compared for their ability to detect HAV Ag under reducing and nonreducing conditions. Application of the 2-ME or DTT treatment procedure to the serologic detection of other viral antigens or viruses whose presence in blood, stool, tissue macerate, or other milieu may be masked by specific antibody appears to be feasible. PMID- 6286868 TI - Monoaminergic mechanisms in stress-induced analgesia. AB - Male albino rats given brief transcutaneous electric footshock showed an elevation in tail-flick response latency--an effect called stress-induced analgesia (SIA). The influence of pretreatments altering monoaminergic neurotransmission on SIA were studied. Inhibition of monoamine storage in presynaptic vesicles reduced SIA, while additional inhibition of serotonin synthesis abolished the SIA response. In contrast, inhibition of serotonin synthesis alone failed to influence SIA. Blockade of dopaminergic receptors reduced SIA, while induction of dopaminergic receptor supersensitivity failed to affect footshock-induced analgesia. Neither inhibition of monoamine oxidase nor blockade of noradrenergic neurotransmission affected SIA significantly. The results suggest that serotonergic and dopaminergic rather than noradrenergic mechanisms contribute to SIA. PMID- 6286867 TI - A behavioural study of the changes in the central nervous system of mice after subchronic treatment with the selective dopamine autoreceptor agonist 3-PPP (dl-3 [3-hydroxyphenyl]-N-n-propylpiperidine). AB - In naive mice the selective dopamine (DA) autoreceptor agonist 3-PPP (dl-3-[3 hydroxyphenyl]-N-n-propylpiperidine) produced a dose-dependent depression of locomotor activity. The duration of action of the depression was short, with no significant depression being noted one or more hours after a dose of 23.47 mg/kg (expressed as the base). Mice, administered the drug twice daily (23.47 mg/kg, in the morning and the evening, i.p.) for 5 days, were, 15 to 25 hours after the last dose, marginally less sensitive to the locomotor depressant effects of a challenge with the same drug. There was no change in the sensitivity of postsynaptic DA and alpha-adrenergic receptors, as assessed by the locomotor stimulant effects of apomorphine and apomorphine plus clonidine, respectively, in reserpine and alpha-methyltyrosine pretreated animals. However, 3-PPP-pretreated mice were most sensitive to the activating effects of d-amphetamine, and this increased sensitivity was blocked by pretreatment with reserpine. In naive mice, a low, DA autoreceptor selective dose of haloperidol (25 micrograms/kg) potentiated the locomotor stimulant effects of d-amphetamine. One explanation for the data obtained is that subchronic pretreatment with 3-PPP produced DA autoreceptor subsensitivity with no concomitant change in postsynaptic DA or alpha-adrenergic receptor sensitivity. The increased sensitivity to d-amphetamine in the 3-PPP pretreated mice may be due to a reduction in the feedback control exerted by the DA released by the d-amphetamine due to the DA autoreceptors having become subsensitive. PMID- 6286869 TI - Alterations in the spontaneous activity of cells in the guinea pig pineal gland and visual system produced by pineal indoles. AB - The indoles serotonin (SER), melatonin (MEL), 5-methoxytryptophol (5-MTL) and 5 hydroxytryptophol (5-HTL) were administered during daytime microelectrophoretically to 240 cells in the pineal gland of the guniea-pig. The action of SER and 5-HTL was predominantly depressant on the electrical activity, MEL and 5-MTL caused an excitation in most of the units. Although MEL and 5-MTL caused fairly similar reactions on average, they appear to act on different cells. The effects of microelectrophoretically applied MEL and 5-MTL on the spontaneous or evoked activity in the visual system (retinal ganglion cells, optic tract, lateral lateral geniculate body, superior colliculus) of the guinea pig were also investigated. Of the 76 cells tested in the visual system 25 of the ON- and OFF-cells increased the rate of discharge when the two indoles were applied. Cells in the optic layer of the superior colliculus showed no measurable response to the application of the two substances. 5-HTL caused no effect on cells in the visual system. PMID- 6286871 TI - Muscarinic cholinoceptors mediate neurally evoked pigment aggregation in glass catfish melanophores. AB - Sympathetic innervation to the melanophores of a siluroid Parasilurus has been the sole instance of such innervation among lower vertebrates, in which the peripheral transmission to the effector cells is peculiarly cholinergic (Fujii and Miyashita, 1976). In an effort to find a similar case, we studied the nature of transmission of melanophores of a glass catfish Kryptopterus. Electrical nervous stimulation brought about melanosome aggregation in the melanophores. While catecholamines were found ineffective, acetylcholine and its analogues were potently active in aggregating pigment. Atropine or scopolamine interferred with the action of both nervous stimulation and acetylcholine. Physostigmine, on the other hand, augmented the cholinergic effects. The conclusion was that the transmission was cholinergic, being mediated by cholinoceptors of muscarinic type, as in the case of Parasilurus. PMID- 6286870 TI - (3H)-muscimol, (3H)-nipecotic acid and (3H)-isoguvacine as autoradiographic markers for GABA neurotransmission. AB - Retinas from rabbit, goldfish and guinea-pig were exposed to (3H) GABA, (3H) nipecotic acid and (3H)-isoguvacine either by intravitreal injection in vivo or by incubations in a balanced salt solution and the distribution of radioactivity was then studied with autoradiography. All substances labelled a similar set of presumed amacrine cells. Incubating at 0 degrees C, in 10-(5)M ouabain, or in 10 (3)M GABA inhibited the labelling by (3H)-muscimol whereas bicuculline (10-(4)M), and glycine (10-(3)M) were less efficient blockers. The result is interpreted as a neuronal uptake of (3H)-muscimol rather than as a GABA receptor binding. All the substances except (3H)-isoguvacine also labelled glia to such a degree that neuronal labelling was often disguised in rabbits and goldfish. Glial labelling by muscimol was less pronounced in guinea-pig. (3H)-isoguvacine (tested only in rabbits) gave a strong labelling of cells with the distribution of GABA neurons and only little glial labelling. PMID- 6286872 TI - Sensitivity in vivo of central alpha 2- and opiate receptors after chronic treatment with various antidepressants. AB - Recent studies suggest that the antidepressant activity could be explained by gradually developing modifications in the sensitivity of some central monoaminergic receptors. As concerns alpha 2-receptors, some largely indirect informations suggest that a subsensitivity develops following chronic treatment with desipramine or imipramine. However it is not clear whether this effect is shared by other antidepressants. In view of the important role of these receptors in the regulation of noradrenergic neuro-transmission, single cell recording and microiontophoretic techniques were used in this work to systematically assess locus coeruleus (L.C.) alpha 2-receptor sensitivity following acute and chronic administration of various antidepressants. The responsiveness of L.C. alpha 2 receptors to iontophoretically applied clonidine was studied simultaneously with the responsiveness of L.C. opiate receptors to iontophoretically applied morphine in order to test the specificity in the putative modifications induced by the antidepressants. The compounds studied were desipramine (DMI), imipramine (IMI), clomipramine (CIMI), zimelidine (ZIM), mianserin (MIAN), iprindole (IPR) and chlorpromazine (CPZ). Long-term treatment with DMI, IMI and ZIM but not with the other clinically effective antidepressant drugs induced a decrease in the responsiveness of L.C. neurons to iontophoretically applied clonidine. None of the drugs tested altered the responsiveness of these neurons to iontophoretically applied morphine. Consequently the therapeutic effectiveness of antidepressant drugs can not be generally related to modulation of the sensitivity of central alpha 2- or opiate receptors. PMID- 6286873 TI - Variability and frequent failure of lucifer yellow to pass between two electrically coupled neurons in Lymnaea stagnalis. AB - The electrically coupled giant neurosecretory neurons VD1 and RPD2 of Lymnaea stagnalis were found to have coupling coefficients ranging from ca. 0.1-0.6. When the fluoroescent dye Lucifer Yellow was injected intracellularly into one of the neurons, in most preparations no dye was observed to pass through into the coupled cell body or the process leading to it. There was no apparent correlation between the amount of dye coupling and the length of time allowed for diffusion of the dye in the cells. In eight preparations, the electrical coupling coefficient was measured before dye was injected. There was no correlation between dye coupling and the electrical coupling coefficient. PMID- 6286874 TI - Complexes of iron and cobalt tetrasulfonated phthalocyanines with apocytochrome c. AB - Artificial cytochromes c have been prepared with Fe(III) and Co(III) tetrasulfonated phthalocyanines in place of heme. Their structure and properties have been investigated by difference spectroscopy, CD, epr, electrophoresis, molecular weight estimation, and potentiometric measurements. The visible absorption spectra show the main peak at 650 nm for the iron compound 685 nm for the cobalt one. It is shown by CD experiments that incorporation of Fe(III)L or Co(III)L into apocytochrome c markedly increases helical content of the protein. Its conformation is, however, significantly altered as compared with the native cytochrome c. The epr and spectroscopic data show that the iron and cobalt phthalocyanine models represent the low spin species with the metal ions in trivalent state. Electrophoresis and molecular weight estimation indicate these complexes to be monomers. Both phthalocyanine complexes have not affinity for additional ligands characteristic for hemoglobin. They react, however, with CO, NO, and CN- when they are reduced with dithionite. Moreover, Co(II)L-apocyt c is able to combine with oxygen suggesting a structural feature in common with the oxygen-carrying heme proteins. Iron(II) complex in the same conditions is oxidized directly to the ferric state. The half-reduction potentials of Fe(III)L apocyt c and Co(III)L-apocyt c are +374 mV and +320 mV, respectively. These complexes are reduced by cytochrome c and cytochrome c reductase (cytochrome bc1). PMID- 6286876 TI - Development of ion metabolism in reaggregated brain cell cultures. AB - Mouse brain cell reaggregates have been used to study changes in sodium- and potassium-dependent ouabain-sensitive adenosine phosphohydrolase (Na+, K+-ATPase) activity and in 86Rb+ uptake and exit during development. Na+, K+-ATPase activity in these cultures has two ouabain-inhibitable components, both of which increased severalfold between day 3 and day 17 in culture. This increase, however, was less than that in developing brain. Little change in either total or extracellular water or in the equilibrium levels of Na+ and K+ occurred during development. The uptake of 86Rb+ measured a 10-min incubation showed only a modest increase during culture, whereas the exit of 86Rb+ from reaggregates preloaded with the tracer increased approximately fourfold. The exit consisted of both K+-independent and K+-stimulated components and the K+-stimulated fraction contributed most of the developmental change. When uptake rates were corrected for the contribution of the developmental changes in exit, these rates were found to increase as well. The 86Rb+ uptake correlated closely with the activity of the Na+,K+-ATPase during development. The pattern of developmental changes in enzyme activity and 86Rb+ uptake and exit suggest that, while little change in the steady-state levels of the ions occurred, the rates of ion movement increase markedly. PMID- 6286875 TI - Eicosanoids: prostaglandins, thromboxanes, leukotrienes, and other derivatives of carbon-20 unsaturated fatty acids. AB - In recent years, knowledge of the biochemistry of oxygenated metabolites of arachidonic acid has greatly increased. Their biological functions in acceleration and prevention of platelet aggregation and in inflammatory and immune reactions are becoming much clearer. The therapeutic value, particularly of PGI2 as well as selective inhibitors of synthesis, is also rapidly advancing. Despite much effort, the functional importance of prostaglandins and thromboxanes in the cNS in normal ongoing physiological processes is still quite uncertain. However, when parenchymal or vascular elements are damaged or invaded by extraneural cells, the synthesis of one or the other member of the eicosanoids is greatly increased and contributes significantly to pathophysiological reactions. Thus, prevention of synthesis is likely to have increasing importance in clinical neurology, particularly in cerebrovascular diseases. PMID- 6286877 TI - Isolation of putative benzodiazepine receptors from rat brain membranes by affinity chromatography. AB - The benzodiazepine receptor from rat brain was solubilised and purified 5200-fold by affinity chromatography. The affinity column contained an immobilized benzodiazepine (delorazepam) and biospecific elution with 6 mM-chlorazepate was achieved. The purified receptor is apparently homogeneous in SDS-polyacrylamide gel electrophoresis. The native protein had a molecular weight of 240,000, and the subunit one of 60,000. The dissociation constant (KD) is 8 nM for [3H]diazepam. A correlation exists between the value of affinity obtained for benzodiazepine derivatives and their known pharmacological effectiveness. PMID- 6286878 TI - Postnatal action of growth and thyroid hormones on the retarded cerebral myelinogenesis of Snell dwarf mice (dw). AB - Snell dwarf mice (dw) showed a lower CNPase activity (59% of the normal controls) only in the cerebrum among different parts of the CNS, and a strikingly reduced level of spontaneous locomotion activity with an indistinct diurnal periodicity in a 24-h record at 40 days of age. Daily administration of bGH and T4 to the dwarfs during the first 40 days of postnatal life restored CNPase activity to the level of the normal controls, and was accompanied by normalization of the pattern of spontaneous locomotion activity. Daily administration of bGH alone also restored CNPase activity and spontaneous locomotion, but to a lesser extent. The daily administration of thyroid stimulating hormone (TSH) alone, however, failed to restore CNPase activity, in spite of the fact that the thyroid glands of the TSH-treated dwarfs were indistinguishable from the normal controls in organization and appearance. These results indicate that the restoration of both the retarded myelinogenesis and abnormal behavior of the Snell dwarf mice might essentially depend upon GH levels and the synergistic effects of T4. PMID- 6286879 TI - GABA autoreceptors are not coupled to benzodiazepine receptors in rat cerebral cortex. AB - Depolarization-induced release of [3H]gamma-aminobutyric acid ([3H]-GABA) from preloaded slices of rat cerebral cortex was inhibited by muscimol and THIP in a dose-dependent fashion. This inhibition of release was prevented by the GABA antagonists bicuculline and picrotoxin. These results confirm previous reports postulating the existence of GABA autoreceptors on GABAergic terminals. Since benzodiazepines are known to facilitate post-synaptic GABA actions, the effect of flunitrazepam on the inhibition of GABA release mediated through the autoreceptors has been examined. At a concentration of 1 microM or 10 microM, flunitrazepam had no effect on the IC50 values for muscimol or THIP in inhibiting stimulated GABA release. It thus seems that GABA autoreceptors are not functionally coupled to benzodiazepine receptors in rat cerebral cortex. PMID- 6286881 TI - Distribution of angiotensin-converting enzyme activity in specific areas of the rat brain stem. AB - Angiotensin-converting enzyme (ACE) activity was measured by a radiochemical assay in 30 specific areas of the rat brain stem. ACE activity is unevenly distributed, with a 60-fold difference between the lowest and the highest activity. The area postrema exhibits the highest activity. The substantia nigra (pars reticulata), the locus coeruleus, the areas A1 and A2, the nuclei commissuralis, and tractus solitarii have a substantial ACE activity, whereas the lowest activity is found in the raphe nuclei and the nuclei of the reticular formation. PMID- 6286880 TI - Endogenous peptides that inhibit brain cyclic AMP phosphodiesterase. AB - Peptide extracts of rat brain powerfully inhibited the cyclic AMP phosphodiesterase activity of rat brain homogenate. Similar extracts of ox brain showed comparable although less potent activity. Preliminary investigation of the physicochemical properties of brain extracts indicated that the rat brain extract contained an active peptide of low molecular weight (about 1400), whereas ox brain contained two such peptides (about 1400 and 900). These studies indicate that endogenous oligopeptides that inhibit cyclic AMP phosphodiesterase are present in brain. Experiments on several pure peptides known to be present in brain. Experiments on several pure peptides known to be present in the CNS showed that the majority were inactive against brain phosphodiesterase, but ACTH(1-24), somatostatin, substance P and Lys8-vasopressin, in descending order of potency, were active. To help distinguish the peptides found in rat and ox brain extracts from known peptides, preliminary analyses of amino acid composition were performed. These suggested that the peptides found in brain extracts were distinct from known peptides having the ability to inhibit cyclic AMP phosphodiesterase. PMID- 6286882 TI - "On" and "off" responses of K+-induced synaptosomal proline release: involvement of the sodium pump. AB - The K+-induced release of the putative neurotransmitter proline in rat cerebral cortex synaptosomes was studied by measuring [5-3H]proline released upon exposure to high-K+ media. In contrast to previous experiments, K+ was not substituted for Na+, which was held constant. Under the resulting conditions of increased osmolarity, two peaks of proline release were obtained, one upon exposure to increased [K+], another upon restoring normal [K+]. The dependence of the latter peak on ionic composition of the medium, and its sensitivity to ouabain are discussed. PMID- 6286883 TI - Effects of cap analogue or cap removal on the translation of rat brain mRNA in vitro. AB - The role of cap structures in the translation of brain mRNA was examined by measuring protein biosynthesis in vitro in wheat germ and reticulocyte systems programmed by mRNA that was either untreated or oxidized by periodate or from which 5'-terminal 7-methylguanosine (m7G) was removed by oxidation and beta elimination. In another series of reactions, amino acid incorporation into polypeptides was measured in the absence and in the presence of varying concentrations of the cap analogue 7-methylguanosine 5'-triphosphate (pppm7G). The results indicated that any of the above treatments interfered with brain mRNA translation, the degree of inhibition depending on the translation system used, the concentration of mRNA, and the source of initiation factors. Homologous brain initiation factors were superior to reticulocyte factors in providing a partial relief from inhibition of translation caused by these treatments. It was also found that synthesis of the brain-specific protein S-100 was inhibited by beta elimination of mRNA, by pppm7G, or by the presence of capped globin mRNA, indicating that the mRNA for this protein was probably capped. PMID- 6286884 TI - Acidosis-induced enhanced activity of the Na-K exchange pump in the in vivo choroid plexus: an ontogenetic analysis of possible role in cerebrospinal fluid pH homeostasis. PMID- 6286885 TI - Avermectin B1a modulation of gamma-aminobutyric acid receptors in rat brain membranes. AB - Avermectin B1a stimulates high-affinity binding of [3H]-gamma-aminobutyric acid (GABA) to receptors in washed rat brain membranes. Scatchard analysis of the data indicates that the drug does not significantly alter the apparent dissociation constant of GABA binding, but increases the detectable number of binding sites from 3.2 to 5.1 pmol/mg protein, (+)-Bicuculline completely blocks control and avermectin B1a-stimulated GABA binding, whereas picrotoxin antagonizes specifically the avermectin B1a-stimulated GABA binding. The avermectin B1a stimulated GABA binding is also chloride ion-dependent, whereas GABA binding in the control is not. These observations suggest that the mechanism of avermectin B1a stimulation of GABA binding may involve the chloride ion channel. PMID- 6286886 TI - Neuroblastoma cell membranes: specificity for cell fusion mediated by a temperature-sensitive mutant of vesicular stomatitis virus. AB - Temperature-sensitive mutant G31 of vesicular stomatitis virus induces mouse neuroblastoma N-18 cells to fuse during infections that are nonpermissive for virus replication, but BHK-21 cells do not undergo the viral glycoprotein mediated cell fusion. The viral glycoprotein was expressed at the cell surface of both N-18 and BHK-21 cells; therefore, the host cell specificity did not stem from an absence of the viral glycoprotein at the surface of BHK-21 cells. Cell fusion readily occurred between infected and uninfected N-18 cells in mixed cultures, demonstrating that the viral glycoprotein was interacting with an uninfected cell for the initial cell-cell interaction of the cell fusion. Mixing infected BHK-21 cells with uninfected N-18 cells resulted in cell fusion initiated by BHK-21 cell-synthesized viral glycoprotein, but 88% of the nuclei in polykaryocytes were N-18 nuclei. The N-18 cell fusion specificity was readily apparent when infected N-18 cells were mixed with uninfected BHK-21 cells; 98% of the nuclei in polykaryocytes were N-18 nuclei. Similar results also were obtained with mixed cultures of N-18 cells and primary astroglial cells. Thus, the viral glycoprotein synthesized in any of the cell types could initiate cell fusion, but the properties of plasma membranes of neuroblastoma cells appeared to be much more suitable for cell-cell fusion. PMID- 6286888 TI - Presence of two benzodiazepine binding sites in the rat hippocampus. AB - Characteristics of receptor binding of diazepam and flunitrazepam in three brain areas were compared. It was found that in the cerebral cortex and cerebellum the number of sites was similar for both ligands and that the affinity of diazepam was four times lower than the affinity of flunitrazepam. In contrast, when binding in the hippocampus was analyzed (assuming the presence of homogeneous binding sites), it was found that the number of binding sites was higher and that the affinity was 17 times lower for diazepam than for flunitrazepam. This difference is due to the presence of two diazepam binding sites in this brain area, as demonstrated by a Scatchard analysis. PMID- 6286887 TI - Alterations in neurotransmitter receptor binding in discrete areas of the copper deficient rat brain. AB - Neonatal copper deficiency produced alterations in central neurotransmitter receptors that were selective with respect both to brain region and to neurotransmitter receptor type. Both high- and low-affinity dopamine receptor densities in the corpus striatum were significantly lowered, 55% and 29%, respectively, when expressed on a wet weight basis. There was a significant decrease in the level of muscarinic receptors in the striatum whether expressed on the basis of wet weight (50%) or protein (27%). A smaller reduction in muscarinic receptor density was observed in the cortex, whereas there was no effect of copper deficiency in the cerebellum. The treatment did not change beta adrenergic receptor binding in either the cortex or cerebellum. The affinities of the receptors for the ligands was not affected by the low-copper diet. It was previously reported that copper deficiency produces regionally specific decreases in the concentrations of dopamine and norepinephrine. The greatest reduction occurred in the concentration of dopamine in the corpus striatum. The results from both studies suggest that copper deficiency in post-weanling rats may induce a selective morphological lesion. PMID- 6286889 TI - Inclusion body myositis. Clinical, biological and ultrastructural study. AB - A 48-year-old patient presented for the past 4 years an amyotrophy of the quadriceps and moderate involvement of the truncal and pelvic girdle muscles. The CK level was elevated (10 times the normal rate) and the EMG revealed a fibrillation pattern on relaxation, myotonic bursts on needle insertion and reduced activity during contraction. The histological study of the muscle biopsy showed nuclear cytoplasmic inclusion bodies and pseudo-myelinic membranes. The case was classified in the inclusion body myositis group. Analysis of the other published cases underlines the variety of the clinical, biological and electromyographical aspects and abnormalities. PMID- 6286890 TI - Spontaneous and evoked electrical discharges from a central demyelinating lesion. AB - Recordings have been made from afferent fibres shown to traverse a focal demyelinating lesion induced in the dorsal columns of the cat by the direct micro injection of lysophosphatidyl choline. Many fibres were spontaneously active, discharging with steady frequencies between 15 and 45 impulses per second or discharging in bursts, for many hours. Small deformations (less than 1 mm) of the spinal cord at the site of the lesion both increased the level of sustained activity, and transiently induced activity in fibres previously electrically silent. The spontaneous and mechanically-induced discharges were shown to proceed both rostrally and caudally from the lesion. The relationship between the experimental observations and the spontaneous and movement-induced sensations experienced by patients with demyelinating lesions of the central nervous system is discussed. PMID- 6286892 TI - Refractory Cushing's disease caused by multinodular ACTH-cell hyperplasia. AB - A patient with pituitary-dependent hypercortisolism, unresponsive to resection of nodules in the anterior lobe, is described. Histochemical stains of the nodules showed multiple, focal, cellular expansions of the fibrovascular stroma. Transitions between normal and expanded adenohypophysial acini were present. Immunoperoxidase stains for ACTH and other pituitary hormones revealed that these multiple foci contained an excess of ACTH-positive cells. Less than 10% of the cells in these foci were negative for ACTH and positive for other hormones. Serial sections showed that these foci of predominantly ACTH-producing acini were not connected. Clinical, morphological, and immunohistochemical data indicated that ACTH-cell hyperplasia caused Crushing's disease in this patient. Pathologic study of individual cases should concentrate on determining whether hyperplasia or adenoma exist at the time of surgical exploration of the pituitary gland, since this determination is important to proper treatment. Tentative criteria to recognize ACTH-cell hyperplasia are: 1. Multiple foci of ACTH laden cells. 2. A minor subpopulation of cells of alternate hormone series. 3. Expansion without destruction of acini in the adenohypophysis. PMID- 6286891 TI - Immunohistochemical detection of factor VIII/von Willebrand factor in hyperplastic endothelial cells in glioblastoma multiforme and mixed glioma sarcoma. AB - The sarcomatous components of most glioma-sarcomas are thought to arise from the neoplastic transformation of hyperplastic endothelial and adventitial vascular cells in a preexisting glioblastoma multiforme. The expression of factor VIII/von Willebrand factor (FVIII/vWF), a marker for endothelial cells, and of glial fibrillary acidic protein (GFAP), a marker for glial cells, was examined in 10 glioblastomas and seven mixed glioma-sarcomas using the peroxidase-antiperoxidase immunohistochemical technique. Hyperplasia of small blood vessels was observed in all 10 glioblastomas; in five, the vascular proliferation had resulted in the formation of prominent glomeruloid structures. FVIII/vWF was detected in the endothelial cells in these vascular structures, but not in the adventitial cells. In the mixed glioma-sarcomas. FVIII/vWF was detected only in endothelial cells; there was no staining for FVIII/vWF in the neoplastic mesenchymal cells. The gliomatous components of the mixed tumors stained intensely for GFAP. These observations indicate that both endothelial and nonendothelial cell types contribute to the small vessel hyperplasia in glioblastomas, and that the sarcomatous components of mixed glioma-sarcomas are derived from either non endothelial cells or endothelial cells that have undergone antigenic loss following transformation. PMID- 6286893 TI - Cyclic AMP-generating systems: regional differences in activation by adrenergic receptors in rat brain. AB - Catecholamine, histamine, and adenosine-mediated accumulations of radioactive cyclic AMP were assessed in adenine-labeled slices from eight rat brain regions. 2-Fluoronorepinephrine, a selective beta-adrenergic agonist, elicited an an accumulation of cyclic AMP in cerebral cortex, cerebellum, hippocampus, striatum, superior colliculi, thalamus, hypothalamus, and medulla-pons. In cerebral cortex and most other brain regions, the beta-adrenergic-mediated response appeared to involve primarily beta 1-adrenergic receptors, while in cerebellum, there was a significant involvement of beta 2-adrenergic receptors. 6-Fluoronorepinephrine, a selective alpha-adrenergic agonist, elicited accumulations of cyclic AMP in all regions except cerebellum. Combinations of the two fluoro derivatives afforded in all brain regions an accumulation of cyclic AMP identical with that elicited by norepinephrine. In hypothalamus, the alpha- and beta-adrenergic responses were significantly greater than additive. In cerebral cortex, the alpha-adrenergic receptor-mediated response appeared to involve alpha 1-adrenergic receptors and to be nearly completely dependent on adenosine, while in other brain regions, the dependence of the alpha-adrenergic response on adenosine was less or absent. Combinations of 6-fluoronorepinephrine and histamine had greater than additive effects in cortex and hippocampus. The results indicate that the interactive control of cyclic AMP-generating systems by alpha-adrenergic, beta-adrenergic, adenosine, and histamine receptors differs significantly among rat brain regions. PMID- 6286894 TI - Differentiation of autonomic neuron precursors in vitro: cholinergic and adrenergic traits in cultured neural crest cells. AB - The development of autonomic neuronal precursors was studied in cultures of microsurgically excised quail neural crest grown alone and associated with other young embryonic tissues. Biochemical differentiation in the cultures was followed by measuring their ability to synthesize acetylcholine (ACh) and catecholamines (CA) from radioactive precursors; cytochemical aspects of their differentiation were examined by techniques including electron microscopy, cholinesterase histochemistry, and CA cytofluorescence. Mesencephalic crest, which can make ACh before explantation, always synthesized ACh after 7 d in culture and often, but not invariably, elaborated small quantities of CA as well. Association with 2-d somite and notochord, 2-d heart, or 4-d hindgut, in medium supplemented with horse serum, resulted in the synthesis of increased amounts of both transmitters. ACh-synthesizing activity was lower and the cholinergic-stimulating effects of somite and heart were abolished in the presence of fetal calf serum. Cervicothoracic (trunk) crest, taken from the level where the dorsal mesoderm is still unsegmented, always produced ACh after culture, but CA was detectable only when the cultures were obtained by initially explanting the entire neural primordium. Co-culture of trunk crest with young embryonic tissue increased ACh synthesizing ability and initiated CA production. Despite their capacity to elaborate neurotransmitter, cultures of either type of neural crest, alone or in association with the above-mentioned tissues, contained very few cells resembling neurons in their phase contrast appearance and none that reacted positively to any of the cytological tests applied. On the other hand, when the sclerotomic moiety of 3-d somite was cultured, trunk neural crest cells that had already migrated into the rudiment in vivo but which had not yet begun to produce detectable amounts of CA underwent rapid differentiation into neurons that synthesized and accumulated large quantities of CA. Stores of CA were detectable cytochemically as early as 24 hr after explantation and the presence of many small, dense core vesicles in neurons and processes was revealed by electron microscopy. ACh-synthesizing activity, demonstrable in freshly dissected sclerotomes, was also present in all of the cultures examined. These results show that (1) during ontogeny, cholinergic traits appear earlier than adrenergic ones in the neuronal precursors contained in the neural crest; (2) some decisive step in the differentiation of the precursor cells of the sympathetic ganglia takes place in vivo within a few hours of the onset of trunk neural crest migration. This coincides with a maturation of the somitic mesenchyme. A similar developmental process does not occur in vitro when 2-d somites and neural crest are associated in histiotypic cultures. PMID- 6286895 TI - Glycine receptor: light microscopic autoradiographic localization with [3H]strychnine. AB - Glycine receptors have been localized by autoradiography in the rat central nervous system (CNS) using [3H]strychnine. The gross distribution of receptors is in excellent accord with the distribution determined by filtration binding assays. Specifically, the density of glycine receptors is greatest in the gray matter of the spinal cord and decreases progressively in regions more rostral in the neuraxis. Glycine receptors were found to be associated with both sensory and motor systems in the CNS. Moreover, there is a striking correlation between areas of high strychnine binding site density and areas in which glycine has been found to be electrophysiologically active. Finally, the anatomic localization of strychnine binding sites may help explain many of the signs and symptoms of strychnine ingestion. For example, individuals consuming subconvulsive doses of strychnine frequently experience altered cutaneous and auditory sensation. We have localized strychnine receptors in areas of the acoustic system known to influence discriminative aspects of audition and in areas of the spinal cord and trigeminal nuclei which modulate discriminative aspects of cutaneous sensation. The alteration of visceral functions (e.g., blood pressure and respiratory rate) associated with strychnine ingestion may be accounted for in a similar manner. PMID- 6286896 TI - Ultrastructural localization and biochemical features of immunoreactive LEU enkephalin in monkey dorsal horn. AB - Leu-enkephalin is an opioid peptide that has been found to modulate nociception in the spinal cord. Both pre- and postsynaptic interactions by enkephalins have been proposed. By the peroxidase . anti-peroxidase immunocytochemical method, we studied the distribution and ultrastructure of neuronal elements in the monkey dorsal horn to elucidate possible morphological substrates for postulated opioid actions. Biochemical analysis of immunoreactive Leu-enkephalin-like peptides in the cord was performed to characterize the forms present in labeled neurons and terminals. At the light microscopic level, fiber immunostaining was found in most areas of gray matter, especially in laminae I to V, and in the dorsolateral funiculus. Cell bodies were located in laminae I, II, III, and V. At the ultrastructural level, in the superficial dorsal horn, we found that neurons with Leu-enkephalin receive numerous types of axon inputs, some of which have been identified previously as originating from the dorsal root. Leu-enkephalin terminals formed primarily axosomatic and axodendritic synapses and less frequently synapsed with other axons. With the same Leu-enkephalin antiserum as used in the immunocytochemistry, a peptide physicochemically similar to intact Leu-enkephalin and two larger Leu-enkephalin-like peptides were identified in monkey spinal cord extracts. It is likely that a family of Leu-enkephalin-like peptides is present in monkey spinal cord and that the labeled elements may contain any or all of these substances. It is concluded that both pre- and postsynaptic physiologic effects of Leu-enkephalin are possible, although the preponderance of axodendritic synapses favors a principal postsynaptic site of action. The anatomical results suggest that neurons containing immunoreactive Leu enkephalin in the dorsal horn, some of which may receive input from primary afferents, modulate nociception by directly synapsing with cells of origin of the spinothalamic tract and also by interacting with primary afferent terminals. PMID- 6286897 TI - Biosynthesis and release of beta-endorphin-, N-acetyl beta-endorphin-, beta endorphin-(1-27)-, and N-acetyl beta-endorphin-(1-27)-like peptides by rat pituitary neurointermediate lobe: beta-endorphin is not further processed by anterior lobe. AB - Continuous labeling and pulse-chase techniques were employed to study the synthesis and secretion of multiple forms of immunoreactive beta-endorphin by cultured dispersed rat anterior lobe cells and intact neurointermediate pituitary lobe. Cell and medium extract immunoreactive beta-endorphin (specific immunoprecipitation and radioimmunoassay) exhibiting a Kav similar to authentic beta-endorphin upon gel filtration was characterized further by nonequilibrium isoelectric focusing, cation exchange chromatography, reverse phase high pressure liquid chromatography, and partial tryptic and chymotryptic mapping. Intact neurointermediate lobes incorporated radiolabeled amino acids into four to six forms of immunoreactive beta-endorphin. Four of these forms were physicochemically similar to authentic beta-endorphin, N-acetylated beta endorphin, beta-endorphin-(1-27), and N-acetylated beta-endorphin-(1-27). Pulse chase studies indicated that a beta-lipotropin-like molecule served as a metabolic intermediate for a beta-endorphin-like molecule. As beta-endorphin-like material accumulated in the cell, some of it was N-acetylated (approximately 18% at 2 hr chase and approximately 65% at 18 hr chase). At later chase times, beta endorphin-(1-27)- and N-acetylated beta-endorphin-(1-27)-like peptides were the predominant molecular species detected. All endorphin forms were detected in unlabeled tissue maintained in culture or tissue continuously labeled for 72 hr and were released into the medium under basal, stimulatory (10(-8) M norepinephrine), or inhibitory (10(-7) M dopamine) incubation conditions. In all cases, beta-endorphin-(1-27)-like species were the predominant forms (more than 70% of total) present in the cells and released into the medium. In contrast, approximately 90% of radiolabeled immunoreactive beta-endorphin extracted from anterior lobe cells and medium similarly incubated appeared to represent the authentic beta-endorphin molecule. Continuous labeling (72 hr) revealed the beta lipotropin/beta-endorphin molar ratio to be approximately 4. We conclude that, in anterior lobe, most of the beta-endorphin is not processed further and is released intact, while in neurointermediate lobe, it serves as a biosynthetic intermediate. PMID- 6286898 TI - The giant serotonergic neuron of Aplysia: a multi-targeted nerve cell. AB - We have examined the various classes of cells that can be innervated by the giant cerebral neuron (GCN), an identified serotonergic cell that functions in arousal and maintenance of feeding behavior. We have found that this single neuron innervates a remarkable variety of postsynaptic targets by means of varicosities bearing active zones. The neuron's presynaptic terminals were identified by electron microscopic radioautography after intrasomatic injection of a tritiated amino sugar precursor of membrane glycoproteins; these are moved to nerve endings by fast axonal transport. In addition to endings on buccal muscle, we have found that GCN forms appositions with the morphological characteristics of synapses on axonal processes and cell bodies of neurons in the buccal ganglion and, unexpectedly, it forms appositions most often with glial cells which form the lining of intraganglionic hemal sinuses. Thus, GCN, through contacts on a variety of postsynaptic targets, has the potential of mediating several different functions, each of which is usually associated with a specific specialized type of neuron. In random electron micrographs, approximately 14% of GCN's varicosities had membrane specializations presumed to be the sites where transmitter is released. In these sections, GCN's active zones were quite small, 0.25 micrometer or approximately five vesicle diameters long. One of GCN's terminals was reconstructed completely from a series of thin sections. It had a single, flat ovoid active zone with an area of 17 micrometers2. We suggest that active zones often are overlooked in random sections of monoaminergic terminals because they are small. PMID- 6286899 TI - Glutamate and aspartate binding sites are enriched in synaptic junctions isolated from rat brain. AB - The binding of the putative excitatory transmitters glutamate (Glu) and aspartate (Asp) was measured in various subcellular fractions in order to assess their degree of localization in synaptic junctions (SJs). For both ligands, specific binding levels increased in the order, whole particulate membranes--crude mitochondrial pellet membranes (P2)--synaptic plasma membranes (SPM), and were highly enriched in SJs, with values approximately 9 times greater than the values in whole particulate membranes. The recovery of binding sites in SJs suggested that the majority of sites in SPMs were junctional in nature. Specific binding sites were found also in other subcellular fractions, such as microsomal membranes, "light" SPMs, and mitochondrial membranes. Sodium ions were able to stimulate the specific binding of both ligands (Asp greater than Glu), the magnitude of the effect between subcellular fractions being in the order, whole particulate membranes greater than P2 greater than SPM. This effect was absent in SJs. Calcium and magnesium ions also enhanced the binding (Glu greater than Asp) in the order, whole particulate membranes greater than or equal to P2 greater than or equal to SPM greater than SJ. The results indicate that Glu and Asp binding sites have a specific synaptic localization and support a role for Glu and Asp receptors in synaptic transmission. PMID- 6286900 TI - The expression of long lasting afterdischarge by isolated Aplysia bag cell neurons. PMID- 6286901 TI - Localization of sodium channels in cultured neural cells. AB - Sodium channels in cultured neural cells were localized by light microscopic autoradiography of specifically bound 125I-scorpion toxin. Ninety percent of the cell-bound 125I-scorpion toxin was associated specifically with sodium channels as assessed by the blocking of autoradiographic labeling by unlabeled scorpion toxin and by depolarization. Sodium channels were distributed uniformly in the surface membrane of neurites and cell bodies of both morphologically differentiated and undifferentiated cells of clone N18 of mouse neuroblastoma C1300. Sodium channels were distributed nonuniformly in many cultured spinal cord neurons. Visual observation indicated that 37 +/- 5% of cultured spinal cord neurons had a higher sodium channel density on the initial segment of one or more neurites than on the cell body. For these neurons, the density on one neurite initial segment averaged 7.4 +/- 1.9-fold greater than on the adjacent cell body. This increased sodium channel density may be the basis of the lower threshold for action potential generation at the axon initial segment of motor neurons and some spinal interneurons in vivo. PMID- 6286902 TI - Presynaptic inhibitory effect of acetylcholine in the hippocampus. AB - (1) In order to investigate the effects of acetylcholine (ACh) on synaptic transmission in the rat hippocampus, extracellular and intracellular recordings were made from pyramidal neurons in an in vitro slice preparation while synaptic inputs to the cell population were stimulated. ACh was applied ionophoretically into somatic and dendritic layers of the slice. (2) ACh applied into the apical dendritic layer of the CA1 region reduced the size of the locally evoked field excitatory postsynaptic potential (EPSP) without altering the size of the afferent fiber volley. Likewise, dendritically applied ACh reduced the size of intracellularly recorded EPSPs. This effect of ACh appeared to be muscarinic since it was not affected by hexamethonium (up to 3 X 10-5 M) but was antagonized by atropine in a dose-dependent manner. (3) The distribution of Ach-sensitive sites matched closely the spatial distribution of activated synapses on the pyramidal cell dendrites as shown by ionophoretic mapping experiments. (4) In contrast to the effects of dendritic applications of ACh, ionophoresis of ACh into the cell layer resulted in an increase and prolongation of EPSPs and a transient decrease in the size of recurrent somatic inhibitory postsynaptic potentials (IPSPs). These effects on synaptic potentials could not be explained by the observed changes in membrane potential and input resistance following somatic application of ACh. (5) Short dendritic applications of ACh had no consistent effect on the membrane potential or slope conductance of pyramidal neurons and did not attenuate the depolarization evoked by brief dendritic applications of glutamate. In addition, the time course of ACh-reduced EPSPs was not different from control. (6) We conclude that ACh exerts a presynaptic inhibitory effect on both excitatory and inhibitory afferents to hippocampal pyramidal neurons. This effect of ACh is widespread, occurring in all regions of Ammon's horn tested as well as in stratum moleculare of fascia dentata. PMID- 6286903 TI - Inhibition of sympathetic preganglionic neurons by catecholamines and clonidine: mediation by an alpha-adrenergic receptor. AB - The neuropil surrounding sympathetic preganglionic neurons (SPNs) receives an abundant catecholaminergic innervation originating from the brain stem. The effect of catecholamines (CA) released at this spinal level on the activity of SPNs is still controversial as is the extent to which this particular CA transmission is affected by central antihypertensive drugs, such as clonidine and alpha-methyldopa. The present study was initiated, therefore, to determine the effects of iontophoretic applications of CAs and clonidine on the discharges of identified SPNs and to determine the type of receptor mediating the action of these compounds. Extracellular recordings were made with five- or six-barrel electrodes in 20 pigeons anesthetized with urethane, artificially ventilated, and immobilized. Data were obtained on 83 SPNs localized in the three first thoracic segments and identified on the basis of constancy of antidromic activation latency and collision. All of the cells sampled were inhibited by the application of low iontophoretic currents of clonidine and by a series of CAs, including alpha-methylnorepinephrine, epinephrine, and phenylephrine. For each compound, the amount of charge necessary to decrease the level of cell firing to 50% of control was calculated. Using this value as an index of drug potency, the following rank order could be determined: clonidine greater than alpha methylnorepinephrine greater than epinephrine greater than norepinephrine greater than phenylephrine. The inhibitory effects of both clonidine and norepinephrine were antagonized by iontophoretic applications of the alpha antagonists, yohimbine, piperoxan, and phentolamine. In contrast, the beta antagonist, sotalol, and the alpha 1 antagonist, prazosin, were found ineffective when similarly applied. It is concluded that CAs and clonidine are inhibitory to the maintained activity of SPNs and that an alpha2-adrenergic receptor may be involved in the action of these compounds. PMID- 6286904 TI - Light microscopic localization of brain opiate receptors: a general autoradiographic method which preserves tissue quality. AB - A general technique is described for using slide-mounted unfixed tissue sections to characterize and visualize drug and neurotransmitter receptors in brain or other tissues. The preparation of material, from fresh frozen, unfixed brain to dried sections securely attached to slides, is described in detail. The tissue can be kept intact during incubation at varying temperatures in solutions containing radiolabeled ligand, ions, buffers, and allosteric effectors. Strategies are described for determining optimal stereospecific binding with highest signal-to-noise ratios and for determining that a "meaningful" receptor is being studied. Dry formaldehyde fixation by vapors from heated paraformaldehyde preserves the tissue quality and traps the ligand near its site on the receptor, permitting subsequent histological processing through alcohols, solvents, and aqueous media, including liquid nuclear track emulsion. Visualization of [3H]naloxone- or [3H]enkephalin-labeled opiate receptor distributions in rat and human brains is achieved by tritium-sensitive film or by classical "wet" emulsion autoradiography. The advantages of the film include its ease of use and the ability to quantify receptor density by densitometry which can be computer-assisted. The advantage of the emulsion is the greater resolution and the concomitant appearance of morphology in cell-stained sections. Examples of correlations of opiate receptor distributions which underlying cytoarchitecture illustrate the potential for receptor localization studies. PMID- 6286905 TI - Spinal cord traumas. Neurobiologic contributions for rehabilitation. PMID- 6286906 TI - Comparison of cold pressor and exercise radionuclide angiocardiography in coronary artery disease. AB - To investigate the role of the cold pressor test (CPT) with radionuclide angiocardiography in the diagnosis of coronary artery disease (CAD), we performed angiocardiography in 52 patients (18 with angiographically normal coronary arteries and 34 with CAD) during the resting state, CPT, and supine bicycle exercise (EX). In normal subjects, left ventricular ejection fraction (EF) was unchanged between rest (58 +/- 9%) and CPT (59 +/- 9%, p = ns), but increased during maximal EX (69 +/- 9%, p less than 0.01). In CAD patients, EF fell from 55 +/- 9% at rest to 49 +/- 9% during CPT (p less than 0.01), and to 53 +/- 11% during EX (p = ns vs. rest). Twenty-seven CAD patients (79%) developed new or worsening areas of dyssynergy during CPT, vs. 25 patients (73%) during EX. Thus, the cold pressor test with radiocardiography appears to be a useful noninvasive test for the diagnosis and functional evaluation of CAD, particularly in patients unable to perform a satisfactory exercise test. PMID- 6286907 TI - The value of 99mTc-pertechnetate brain scan as a screening for chronic subdural hematoma in elderly patients. PMID- 6286908 TI - Effect of dietary or genetic copper deficiency on brain catecholamines, trace metals and enzymes in mice and rats. AB - Previous studies by others indicated that alterations in brain catecholamines were different for perinatal copper deficiency produced by diet in rats and that resulting from a genetic mutation of the X-chromosome, Menkes' syndrome in humans and brindled mice. Thus, copper deficiency was studied in a model in which dietary and genetic deficiency (brindled mice) were compared in two strains of the same species. C57BL and C3H/HeJ mice. Dietary copper deficiency was also produced in rats for comparison. In brain, both dietary and genetic copper deficiency resulted in impaired growth, low brain copper levels, greatly decreased norepinephrine concentrations but normal dopamine levels. The activity of brain cytochrome oxidase was greatly depressed following both dietary and genetic copper deficiency, suggesting a functional deficit of copper. However, the activity of another cuproenzyme, dopamine-beta-hydroxylase, was significantly elevated in deficient animals. The elevation was observed when either copper or N ethylmaleimide was added to inactivate an endogenous inhibitor. The cause of low brain norepinephrine remains unknown; however, depressed brain norepinephrine may be partly responsible for functional changes in the deficient animals, such as hypomyelination, since the activity of the myelin protein, 2',3'-cyclic nucleotide 3'-phosphodiesterase, was lower in the most deficient animals. PMID- 6286909 TI - Effects of fiber on digestibility and transit time in dogs. AB - This study examines effects of variations in fiber content on nutrient assimilation, fecal output, and gastrointestinal transit time in the dog. Four normal Beagles were fed four diets in a randomized block design. The basal diet was a canned, balanced, meat-based dog food (Alpo Trio) to which added 3, 6, and 9% by weight of alpha cellulose (Solka Flok). Food intake and fecal outputs were recorded for 5-day periods. Samples of diets and fecal collections were analyzed for dry matter, nitrogen, fat, carbohydrate and ash; digestibilities were calculated. Transit times were measured by a radiographic marker technique. Fecal weight and water increased linearly; digestibility of dry matter decreased from 90 to 70% and ash from 43 to 32% with added fiber. Responses of protein, carbohydrate and fat were less pronounced but were regular; regressions of their digestibilities on added fiber were significant. Regression estimate of true digestibility for alpha-cellulose was 6%. Intestinal transit time decreased from a mean of 37.4 to 28.7 hours with added fiber. Decreased intestinal time would contribute to depression of fry matter digestibility. Increased fecal water output probably also reflected retention by fiber. PMID- 6286911 TI - NADPH-dependent reduction of cytochrome P-450 in liver microsomes from vitamin C deficient guinea pigs: effect of benzphetamine. AB - The effect of dietary vitamin C on the reduction of cytochrome c and cytochrome P 450 by NADPH-cytochrome P-450 reductase was determined in guinea pig liver microsomes. Acute vitamin C deficiency decreased hepatic microsomal cytochrome P 450 content 21% and the rate of cytochrome P-450 reduction 66% without affecting cytochrome c reduction. However, the vitamin status of the animals did not affect the enhancement in cytochrome P-450 reduction produced by the addition of a type I substrate to the reaction mixture. The results suggest that vitamin C deficiency selectivity affects the transfer of electron from NADPH to cytochrome P-450 and that this effect does not result from a change in the spin state of cytochrome P-450. PMID- 6286910 TI - Cholecalciferol requirements of young turkeys under normal conditions and during recovery from rickets. AB - Day-old turkeys fed vitamin D-deficient diets became rachitic within 17-24 days. The symptoms included reductions in body weight, plasma calcium and inorganic phosphorus, plasma and intestinal calcium-binding protein (CaBP), plasma 25 hydroxycholecalciferol [25(OH)D3], bone ash, and kidney 25(OH)D3-24-hydroxylase and a rise in kidney 25(OH)D3-1-hydroxylase activity. Supplementation of the diet with 12.5 micrograms cholecalciferol per kilogram was sufficient to promote maximal body weight and normal plasma calcium, Plasma calcium, plasma phosphorus and bone ash. Feeding diets containing 250 or 1250 micrograms cholecalciferol per kilogram resulted in a reduced body weight. An increased in the concentration of plasma 25(OH)D3 with increasing dietary cholecalciferol concentration was observed. Feeding vitamin D-deficient rachitic birds for 4 days a diet containing 50 micrograms cholecalciferol per kilogram restored plasma calcium and phosphorus and bone ash. Body weight remained lower than that of the control for an additional 6-day period. Additional cholecalciferol, 25(OH)D3 or 1 alpha hydroxycholecalciferol in the diet, intramuscular injection of the vitamin D derivatives, or a high-calcium, high-phosphorus diet did not accelerate the recovery from the rachitic state. PMID- 6286912 TI - Evidence for production of an inactive collagenase by fibroblasts from phenytoin enlarged human gingivae. AB - When measured by radioimmunoassay, fibroblasts derived from the overgrown gingivae of phenytoin-treated epileptic individuals synthesize and release elevated amounts of collagenase in vitro, as compared to similar-appearing fibroblasts from normal, non-phenytoin-treated persons. However, it appears that much of the immunoreactive enzyme is unable to degrade reconstituted collagen in culture. This preliminary finding in 9 different strains of cells indicates that reduced collagenase activity by a subpopulation of cells may contribute to the development of phenytoin-induced gingival overgrowth. PMID- 6286913 TI - Coxsackie B4 infection and islet cell antibodies three years before overt diabetes. PMID- 6286914 TI - Persistence of cytomegalovirus in human milk after storage. PMID- 6286915 TI - Human beta-endorphin-like immunoreactivity in the perinatal/neonatal period. PMID- 6286917 TI - Bilateral Wilms' tumor and secondary malignancies. AB - A series of six patients with bilateral Wilms' tumor (nephroblastoma) is presented. Multimodal therapy yielded a survival rate of 83% at 2 yr of follow up. However, at the end of a later follow-up period only two patients (33%) were alive. of the 4 patients who died. Only 1 died of Wilms' tumor. One died of complications of aggressive chemotherapy and two patients died after 12 and 16 yr following treatment of secondary malignant tumors arising in the irradiated region. Patients with bilateral tumor should be followed at regular intervals for the duration of life for the occurrence of secondary malignant tumors. PMID- 6286916 TI - Treatment of bilateral Wilms' tumors--a 22-yr experience. AB - A total of 157 children with Wilms' tumors were treated at the Children's Memorial Hospital during the years 1956-1978. Fourteen (or 9%) of these children had bilateral lesions. During the early years, treatment consisted of surgical excision of one kidney with radiation to the second tumor. During the second decade of this study a more aggressive approach to bilateral tumors was used consisting of nephrectomy on the side of the large tumor, with a heminephrectomy on the contralateral side and chemotherapy. More recently, we have attempted bilateral heminephrectomy when possible, together with increasingly aggressive chemotherapy. This series of patients allows us to compare the results of various treatment modalities among our own patients as well as those reported from other pediatric surgical centers. PMID- 6286918 TI - [Adsorption of cetylpyridinium chloride to hydroxyapatite (author's transl)]. PMID- 6286919 TI - [Regulation of chronic proliferative inflammation by anti-inflammatory drugs (author's transl)]. PMID- 6286920 TI - Inhibition of calmodulin by phenothiazines and related drugs: structure-activity relationships. PMID- 6286921 TI - Pharmacological studies on sympathoinhibition produced by the medial medullary depressor region: proposed gamma-aminobutyric acid involvement in inhibition of somatosympathetic reflexes. PMID- 6286923 TI - Characteristics of adrenoceptors in a nociceptive pathway in the mesencephalic reticular formation of the rat. AB - Norepinephrine (NE) may be a neurotransmitter in a nociceptive pathway in the mesencephalic reticular formation (MRF) of the rat. When NE is applied microiontophoretically to certain neurons in rat MRF, it produces a change in neuronal firing rate similar to that produced by a noxious stimulus (foot pinch) in these neurons. If NE is a neurotransmitter involved in the transmission of nociceptive information in the MRF, specific adrenergic receptor antagonists, administered microiontophoretically, should block both the firing evoked by NE, administered microiontophoretically, and the firing evoked by foot pinch in these neurons. When phentolamine (an alpha adrenoceptor antagonist) was administered microiontophoretically it blocked the increases in neuronal firing rate evoked either by foot pinch or by NE. In contrast, propranolol (a beta adrenoceptor antagonist) rarely blocked the firing evoked by either foot pinch or by the microiontophoretic administration of NE. Thus, it appears that if NE is involved in the transmission of nociceptive information in the MRF of the rat, this information transfer is mediated by alpha adrenergic receptors. PMID- 6286922 TI - Effect of treatment of rats with antidepressants on melatonin concentrations in the pineal gland and serum. AB - The effect of administration of antidepressant drugs to rats on catecholamine induced rises in the concentration of melatonin in the pineal gland and serum was studied. Increases in melatonin levels were produced by either the exogenous administration of different doses of L-isoproterenol or by exposure of rats to darkness. Repeated administration of either desmethylimipramine or nialamide reduced significantly the elevation in melatonin concentrations in the pineal gland produced by either L-isoproterenol or darkness. The rise in serum melatonin seen at night was also blunted significantly in rats treated repeatedly with these drugs. By contrast, the concentration of melatonin in either the pineal gland or serum was not reduced in rats given desmethylimipramine or nialamide acutely. In fact, daytime concentrations of melatonin both in the gland and in serum were significantly increased after either acute or repeated treatment with nialamide. The nialamide-induced increase in daytime concentrations of melatonin in the pineal gland was prevented if the rats were treated with propranolol. The diminished hormonal responsiveness produced by the repeated administration of antidepressants occurred at a time when the binding of [3H]dihydroalprenolol to pineal gland homogenates was reduced. Thus, a hormonal response to catecholamines regulated via beta adrenergic receptor activation is reduced by repeated antidepressant treatments in conjunction with the development of beta adrenergic receptor subsensitivity. PMID- 6286924 TI - Influence of adrenocorticotropin hormone and yohimbine on antidepressant-induced declines in rat brain neurotransmitter receptor binding and function. PMID- 6286925 TI - 3,4-diaminopyridine alters acetylcholine metabolism and behavior during hypoxia. AB - Low oxygen (hypoxia) decreases the release and synthesis of acetylcholine (ACh) as well as behavioral performance (i.e., tight rope test scores). These hypoxic induced impairments are partially reversed by 3,4-diaminopyridine (3,4-DAP). With mouse brain slices, the Ca++-dependent-K+-stimulated synthesis of ACh from [U 14C]glucose decreased 45% under 2.5% oxygen and 80% under 0% oxygen. During hypoxia, 3,4-DAP (10 nM) increased in vitro ACh synthesis 67% (2.5% oxygen) and 63% (0% oxygen). The Ca++-dependent-K+-stimulated release of ACh declined by 55 or 67% under 2.5 or 0% oxygen, respectively.3,4-DAP partially reversed this hypoxic impaired release of ACh under 2.5 (+69%) or 0% (+226%) oxygen. In vivo, chemical hypoxia (i.e., NaNO2-induced methemoglobinemia) reduced ACh synthesis from [U-14C]glucose in striatum (-82%), cortex (-68%) and hippocampus (-55%). During hypoxia, 3,4-DAP (10 pmol/kg) increased [U-14C]glucose incorporation into ACh in the hippocampus (+62%) and striatum (+36%). Tight rope test performance decreased 89% during hypoxia and 3,4-DAP diminished this decline to only 55%. These findings demonstrate that the hypoxic-induced deficits in ACh metabolism and behavior can be partially reversed by interaction with Ca++ homeostasis. The therapeutic usefulness of 3,4-DAP in the treatment of human metabolic encephalopathies remains to be evaluated. PMID- 6286926 TI - Mechanism of the clonidine-induced protection against acetylcholinesterase inhibitor toxicity. AB - Clonidine, an alpha-2 adrenergic agonist can inhibit the release of acetylcholine from central and peripheral cholinergic neurons. This study was designed to examine the ability of clonidine to protect animals from the toxic manifestations of cholinesterase poisoning. Physostigmine, a central and peripheral cholinesterase inhibitor produced tremors, and at high doses death, by respiratory paralysis. Mice were injected with physostigmine at a dose (0.75 mg/kg) which evoked tremors in 100% and death in 90% of the animals. Clonidine pretreatment (0.3 mg/kg) increased the onset latency to tremor from 5 to 20 min, increased the onset latency to death from 12 to 24 min and increased the percentage of survivors to 50%. Yohimbine (1 mg/kg) reversed these protective effects of clonidine. The physostigmine-induced accumulation of forebrain and hindbrain acetylcholine also was reduced by 50% in both brain regions in clonidine-pretreated mice. Neostigmine, a selective peripheral cholinesterase inhibitor, induced respiratory paralysis which was not affected by clonidine pretreatment. These findings indicate that central cholinergic neurons involved in the regulation of respiration and fine motor control, but not peripheral motor neurons, are inhibited by clonidine acting on alpha receptors. PMID- 6286927 TI - Solubilization and characterization of active opiate binding sites from mammalian brain. AB - Active binding sites have been solubilized from cell membranes derived from mammalian brain. High affinity, stereospecific binding to soluble sites was demonstrable when membranes from rat brain, human frontal cortex and bovine corpus striatum were treated with digitonin or glycodeoxycholate, provided that the binding assay was conducted at 25 degrees C or below and in the presence of 50 to 100 mM NaCl. The yield of solubilized binding sites extracted from brain cell membranes was increased substantially (up to 43% yield from bovine striatum) when membranes were treated with detergent solutions containing 0.5 to 1.0 M NaCl. This effect was not observed when LiCl, KCl or (NH4)2SO4 were substituted for NaCl. Evidence for the solubility of the binding sites was provided by two criteria: nonsedimentation after 2 hr of centrifugation at 10(5) X g and an apparent molecular weight of 3 to 4 X 10(5) as determined by gel filtration. PMID- 6286928 TI - Biochemical evidence for pulmonary endothelial cell injury after carbon tetrachloride administration in mice. AB - Angiotensin converting enzyme (ACE) is concentrated in pulmonary endothelial cells. Its release into the general circulation after lung damage may provide an index of pulmonary microvascular injury. In the present study, we have examined the effect of the volatile solvent carbon tetrachloride on serum levels of ACE. In mice, a dose-response relationship was obtained in serum ACE 24 hr after acute oral doses of carbon tetrachloride ranging from 0.1 to 2.8 ml/kg. Serum levels of ACE returned to normal by 48 hr after treatment. The increase in serum ACE after carbon tetrachloride treatment correlates with a significant reduction in lung ACE, whereas liver and kidney levels are unaffected. The elevation in serum ACE which occurs after carbon tetrachloride would appear to be, therefore, the result of release from the lungs. Because tissue injury is associated with inflammation, we investigated what effect an anti-inflammatory drug has on the elevation of serum ACE by carbon tetrachloride. Pretreatment of mice with 100 mg/kg of aspirin, orally, produced a 53% reduction in the elevation of serum ACE produced by carbon tetrachloride. These studies strongly suggest that carbon tetrachloride can produce damage to pulmonary endothelial cells. A model to explain the transient nature of elevated serum ACE based on current knowledge of glycoprotein structure and metabolism is proposed. PMID- 6286929 TI - Stereochemical requirements of alpha-2 adrenergic receptors. PMID- 6286930 TI - Effect of increments in norepinephrine concentrations on seizure intensity in the genetically epilepsy-prone rat. PMID- 6286932 TI - Mechanism of action of quinidine on heart rate in the dog. AB - The known vagolytic effect of quinidine has been termed "atropine-like." This effect was studied in vitro and in vivo. Unlike atropine, quinidine was not found to exert any competitive antagonism to acetylcholine at the muscarine-sensitive cholinoceptors of isolated guinea-pig atria. In the narcosed dog subjected to vagal stimulation, the effects of slow perfusion of quinidine (0.5 mg/kg/min for 120 min) were also different in several ways from those of atropine. The tachycardia induced by quinidine was not concomitant with an inhibition of the cardiomoderation caused by vagal stimulation. Tachycardia appeared at low plasma levels, whereas reduction of vagal influence only occurred at high concentrations. Even then, the effects of vagal stimulation were never totally suppressed. After propranolol, quinidine had no tachycardiac action. This could not be attributed to any difference in metabolism because plasma levels of quinidine were the same in dogs pretreated or not with propranolol. Evidence reported here suggests that the cardioacceleration induced by quinidine is not due to a direct vagolytic action. The slight lowering of vascular resistance and arterial pressure observed cannot fully account for this effect either. An adrenergic mechanism involving beta receptors seems likely. In any case, to describe the effects of quinidine on heart rate as atropine-like is evidently inappropriate and misleading. PMID- 6286931 TI - Actions of GABAergic agents on dopamine metabolism in the nigrostriatal pathway of the rat. AB - The gamma-aminobutyric acid (GABA) transaminase inhibitor aminoxyacetic acid, diazepam and the GABA agonists muscimol, 4,5,6,7-tetrahydroisoxazolo-[5,4-C] pyridin-3-ol, kojic amine and SL-76002 all depressed nigrostriatal dopamine release as measured by decreased 3-methoxytyramine after parenteral administration. Experiments with intrastriatal and intranigral injections as well as parenteral injections in acutely hemisected and kainate lesioned animals demonstrated that parenteral muscimol inhibits the nigrostriatal pathway at the level of the substantia nigra. In contrast, parenteral 4,5,6,7 tetrahydroisoxazolo-[5,4-C]-pyridin-3-ol, kojic amine and aminoxyacetic acid inhibit the nigrostriatal pathway acting both within the substantia nigra and on presynaptic nigrostriatal GABA receptors in the striatum. A unique interneuronal GABA receptor also appears to be present in the striatum which is activated by intrastriatal muscimol but not 4,5,6,7-tetrahydroisoxazolo-[5,4-C]-pyridin-3-ol or kojic amine. Activation of these receptors leads to increased dopamine release in the striatum; however, this receptor population does not appear to be stimulated after parenteral administration of GABAergic agents. In summary, multiple GABA receptors appear to be present within the central nervous system which possess differential sensitivities to the GABA agonists examined; however, after parenteral administration, all GABAergics appear to depress nigrostriatal dopaminergic transmission. PMID- 6286933 TI - Effect of diltiazem, a calcium antagonist, on myocardial pH in ischemic canine heart. AB - The effect of diltiazem on ischemic myocardial acidosis was studied in the canine heart whose left anterior descending coronary artery was partially occluded to reduce coronary flow to about one-third (partial occlusion). Myocardial pH was measured by use of a micro glass pH electrode. The pH decreased from 7.53 to 6.93 by partial occlusion and appeared to reach a steady state within 30 min. Saline or drugs were injected i.v. 30 min after partial occlusion. The decreased pH increased spontaneously by 34% of the total reduction of pH in the next 60 min after saline injection. Diltiazem (100 micrograms/kg) potentiated the increase in pH; the pH increased by 76% 60 min after the injection. Propranolol (1 mg/kg) also potentiated the increase in pH that had been decreased by partial occlusion. The relation between pH decrease and the tissue levels of metabolites was also studied. The reduction of myocardial pH from 7.5 to 6.8 was accompanied by a 2 fold increase in the tissue lactate content and by decreases in the ATP and creatine phosphate contents. There was a significant correlation between the hydrogen ion concentration calculated from the pH and each of the lactate, ATP and creatine phosphate contents. The present study indicates that diltiazem attenuates ischemic myocardial acidosis. PMID- 6286934 TI - Central vasopressor responses to conjugated estrogens in rats may be mediated via a renin-angiotensin system in the brain. AB - Central effects of sex steroids on cardiovascular regulation and on behavior were investigated by injecting conjugated estrogens i.c.v. in conscious or anesthetized rats. Conjugated estrogens (0.1-10 micrograms) produced dose-related increases in pressure and behavioral excitation in all rats and drinking in some rats. Marked increases ii pressure and excitation with tonic convulsions resulting in death were produced by similar injections of conjugated estrogens (100 micrograms). Pretreatment with angiotensin I converting enzyme inhibitor or angiotensin II analog abolished both vasopressor responses and behavioral excitation in conscious rats. When rats were later anesthetized with urethane to allow recording of sympathetic nerve activity, 10 and 100 micrograms of conjugated estrogens produced dose-related vasopressor responses accompanied by corresponding increases in peripheral sympathetic nerve activity. Again, converting enzyme inhibitor or angiotensin II analog abolished the increases in both blood pressure and sympathetic nerve activity. By contrast, when conjugated estrogens were injected i.v. up to 2 mg/kg, neither blood pressure nor sympathetic outflow was affected. Collectively, our results suggest that conjugated estrogens infused centrally activate a renin-angiotensin system in the brain, which results in sympathetic hyperactivity leading to blood pressure elevation. PMID- 6286935 TI - Termination of responses to sympathetic nerve stimulation and to noradrenaline in a perfused arterial preparation: the role of neuronal and extraneuronal uptake. AB - Perfused branch vessels of bovine radial artery in vitro responded to sympathetic nerve stimulation and to noradrenaline added extralumenally and intralumenally with constrictions monitored as flow reductions. Cocaine and 17 beta-estradiol, inhibitors of neuronal and extraneuronal uptake, respectively, enhanced responses to exogenous noradrenaline given by either route. An analysis of response recovery rates, namely termination of action, revealed that the extraneuronal and neuronal uptake process each accounted for between 35 to 50% of the total inactivation capacity of the perfused neuroeffector system for noradrenaline. Responses to periarterial nerve stimulation over a 0.5 to 10 Hz frequency range, involving minimal to moderate flow reductions, were substantially magnified by cocaine and by 17 beta-estradiol. An analysis of response duration and recovery rates showed that extraneuronal processes were more significant than neuronal uptake in terminating noradrenergic transmitter action over the frequency band of 0.5 to 10 Hz. PMID- 6286936 TI - Evidence for the presynaptic localization of a high affinity opiate binding site on dopamine neurons in the pedal ganglia of Mytilus edulis (Bivalvia). AB - Considerable evidence has demonstrated an interrelationship between the enkephalinergic and dopaminergic systems in both the mammalian and invertebrate nervous systems. We have described recently the presence of two classes of high affinity opiate binding sites in the nervous tissue of the marine mollusc Mytilus edulis. In order to examine the physiological role of these high affinity opiate sites, M. edulis pedal ganglia (Pg) were treated with the selective neurotoxin 6 hydroxydopamine (1 micrograms/animal, applied topically to the intact Pg); animals were sacrificed 5 days after treatment. The dopamine content of the Pg from lesioned animals was reduced to 33% relative to that of Pg from control animals. Neither serotonin nor norepinephrine levels were reduced. Fluorescent micrographs of formaldehyde-treated Pg from both lesioned and control animals revealed that the neurotoxic substance accumulates in the synaptically rich neuropil and not in the cortex of the Pg. Thus, the partial reduction in dopamine levels may reflect nearly total loss of dopamine in terminals with essentially no change in the nerve cell bodies. High affinity binding of the potent opioid peptide 125I-labeled FK 33-824 (2 nM) was reduced by 81% and low affinity binding (10 nM peptide) by 43% in Pg from lesioned animals relative to that in control tissue. In addition, D-Ala2-Met5-enkephalin, beta-endorphin and etorphine failed to change dopamine levels in lesioned animals. Together, these results suggest that the high affinity opiate binding sites that mediate alteration in dopamine levels are on dopaminergic presynaptic terminals. PMID- 6286937 TI - Inhibition of respiratory neural discharges by clonidine and 5-hydroxytryptophan. AB - Activation of receptors for norepinephrine or serotonin in the central nervous system by i.v. injection of clonidine (10-50 micrograms/kg) or 5 hydroxytryptophan (20-40 mg/kg) inhibits phrenic neural discharges in anesthetized, artificially ventilated cats. Clonidine induces a rapid and complete inhibition of phrenic nerve activity which lasts for 1 to 3.2 hr. The inhibition is prevented by prior administration of phenoxybenzamine (10 mg/kg) or tolazoline (3 mg/kg). 5-Hydroxytryptophan, injected after inhibition of peripheral amino acid decarboxylase (carbidopa, 30-50 mg/kg), elicits a gradual but complete inhibition of phrenic nerve discharges which persists for 1 to 10 hr and is unaltered by alpha or beta adrenoceptor blocking agents. The inhibitions produced by clonidine and 5-hydroxytryptophan are overcome transiently during hypercapnia. Stimulation of carotid body chemoreceptors by i.a. injections of lobeline, doxapram or 0.015 N HCl in saline also briefly reinstates phrenic nerve discharges after inhibition by clonidine. Inhibition is also overcome during electrical stimulation of the carotid sinus nerve. PMID- 6286938 TI - The relation between tonicity and impulse-evoked transmitter release in the frog. AB - 1. The increase in miniature end-plate potential (m.e.p.p.) frequency in response to tetanic stimulation of the motor nerve at frog neuromuscular junctions in Ca(2+)-free, Mg(2+) EGTA-containing (0 Ca(2+)-Mg(2+) EGTA) solutions of varying tonicity has been studied. The response to stimulation is markedly increased in hypertonic solutions and is decreased in hypotonic solutions. Under these conditions changes in tonicity have comparable effects on stimulated and spontaneous quantal release.2. The tonicity was raised by adding sucrose, NaCl or glycine to the extracellular solution. The effects of the addition depended primarily on the increase in osmolarity of the solution, not on the chemical species producing it.3. The tonicity was decreased by lowering [NaCl](o). The hypotonic solution decreased the response to tetanic stimulation. When the tonicity of the solution with the low [NaCl](o) was restored to normal by adding sucrose, the response was restored to its usual level. These results suggest that in 0 Ca(2+)-Mg(2+) EGTA solutions stimulation does not enhance the probability of quantal release by raising [Na(+)](i).4. Repeated bouts of tetanic stimulation produced almost identical responses. In some instances the frequency continued to rise after the end of the tetanic stimulation, as reported by Erulkar & Rahamimoff (1978). This suggests that the stimulation of the nerve leads to the elevation within the terminal of a substance that in turn liberates an activator for quantal release.5. The Q(10) for the increase in probability of quantal release is as high as 7. High Q(10) values have also been reported for spontaneous m.e.p.p. frequencies. Tonicity and temperature appear to affect spontaneous and stimulated quantal release similarly. PMID- 6286939 TI - Mechanisms of transmembrane calcium movement in cultured chick embryo ventricular cells. AB - 1. Uptake of calcium was studied in spontaneously contracting monolayers of cultured chick embryo ventricular cells. Ca exchange could be separated into two components: a rapid phase with a rate constant of 3.91/min, accounting for 1.6 nmol/mg protein; and a slower phase with a rate constant of 0.069/min, accounting for 2.7 nmol/mg protein.2. Negatively inotropic concentrations of the slow Ca channel blocker verapamil inhibited the rapid phase of Ca uptake partially, with a maximum inhibition of 30-40% observed at concentrations of verapamil which completely inhibited contraction.3. The component of Ca uptake not inhibited by verapamil could be stimulated up to 25-fold by elevation of intracellular Na concentration and reduction of extracellular Na concentration, and thus appeared to represent at least in part Ca uptake via Na-Ca exchange.4. Ca uptake by cultured cells could be almost completely inhibited by exposure to LaCl(3), 1 mmol/l, within 5 s. This same concentration of La completely inhibited contraction within 5 s.5. During efflux of (45)Ca from cells, exposure to La (1 mmol/l) slightly inhibited efflux initially with more marked inhibition of Ca influx after 3 min of La exposure. There was no evidence for a component of superficial La-displaceable Ca, and thus the rapid phase of Ca uptake probably is due to an intracellular rapidly exchanging Ca pool. Efflux of Ca from this rapidly exchanging intracellular Ca pool was not significantly altered by exposure to Na-free choline chloride solutions.6. We conclude that rapid Ca uptake in these cultured myocardial cells is due primarily to Ca influx via the slow Ca channel and via Na-Ca exchange. In the presence of physiological [Ca(2+)](i), efflux of Ca from this intracellular Ca pool does not appear to be due to Na-Ca exchange. PMID- 6286940 TI - Fluoride blocks an inactivation step of transduction in a locust photoreceptor. AB - 1. Photoreceptors in a superfused retina of a locust compound eye are treated with saline containing 10 mM-NaF, while their intracellular resting potential and responses are recorded using glass micropipettes.2. Treatment for two minutes with 10 mM-NaF, followed by a series of brief, bright flashes of light, results in an irreversible, noisy depolarization of approximately 10 mV. The final, stable level of depolarization is reached through the summed effect of each of the noisy, depolarizing afterpotentials that follow every response of the cell to a light flash. If kept in darkness after treatment with NaF, the noisy depolarization still develops, but more gradually, over a period of 5 min.3. The voltage noise induced by NaF mimics light-induced voltage noise when the two are compared at mean depolarizations of more than 15 mV. At very small depolarizations, however, fluoride-induced noise cannot be resolved into the large discrete events (bumps) that are typical of the response of a dark-adapted photoreceptor to a single photon.4. The complete replacement of the superfusate sodium by choline reversibly reduces the fluoride-induced noise and depolarization to the same extent as it does the light-induced noise and depolarization of an illuminated cell.5. Increasing the superfusate calcium concentration from 0.5 to 10 mM also reversibly reduces fluoride-induced noise and depolarization to the same extent as it does light-induced noise and depolarization. This action of calcium is accompanied by an increase in a cell's input resistance which opposes the reduction caused by light or fluoride treatment.6. The results confirm the proposal (Payne, 1981) that anionic metabolic inhibitors cause spontaneous activity in sodium channels that are normally opened by light. A model is proposed in which fluoride acts by blocking the inactivation of a late stage in the transduction mechanism. PMID- 6286941 TI - Acetylcholine measured at short time intervals during transmission of nerve impulses in the electric organ of Torpedo. AB - 1. The amounts of total acetylcholine (ACh) and ATP, and of vesicle-bound ACh were measured at short time intervals in the electrogenic tissue of Torpedo marmorata. The aim of this study is to approach with biochemical analysis the speed of electrophysiological phenomena.2. A stimulator coupled to a rapid freezer device was used to quench a number of tissue samples simultaneously, at different time intervals during transmission of a brief train of impulses at 100 Hz.3. The level of total ACh decreased significantly with the first ten impulses. Then a rapid but transient increase in total ACh occurred, reaching a maximum value by the fifteenth to sixteenth impulse.4. Vesicle-bound ACh did not exhibit any changes parallel to those of total ACh, and did not decrease beyond the control level during transmission of twenty impulses at 100 Hz.5. The amount of ATP in the tissue varied in close relation to that of total ACh. No significant phase shift was observed between the transmitter and the nucleotide and the ACh/ATP molar ratio was not significantly different from 1.6. The shortest time interval investigated in this work was 10 ms. The rate at which the pieces of tissue are quenched for biochemical measurements when plunged into a liquid at low temperature has been estimated. It has also been evaluated to what extent the freezing rate may distort measurements of the biochemical changes occurring in the tissue.7. It is concluded that fast freezing appears to be a valuable approach for investigating the rapid biochemical changes underlying cholinergic transmission; a better time resolution might be reached at the price, however, of greatly reducing the size of the samples. The second conclusion is that transmission of a brief train of impulses is accompanied by significant changes in the amount of extravesicular ACh. PMID- 6286942 TI - Acetylcholine changes underlying transmission of a single nerve impulse in the presence of 4-aminopyridine in Torpedo. AB - 1. Transmission of a single nerve impulse has been investigated at the nerve electroplaque junction of Torpedo marmorata in the presence of 4-aminopyridine (4 AP), a drug which powerfully potentiates evoked transmitter release.2. Three methodological approaches were used conjointly. These were (i) electrophysiological recording of the compound electroplaque potential (e.p.p.), (ii) radiochemical measurement of evoked acetylcholine (ACh) release and (iii) analysis of the content of ACh and ATP in the tissue at brief time intervals during the course of the e.p.p. and soon after. The last was achieved by using a stimulator coupled to a rapid tissue freezer.3. In the response to a single stimulus, 4-AP enhanced in a dose-dependent manner the size of the e.p.p., increasing the duration much more than the amplitude. At 10(-4) M-4-AP, this resulted in the generation of a characteristic ;giant e.p.p.' whose area (in V x ms) was approximately 120 times greater than that of a normal e.p.p.4. The giant e.p.p. consisted of an initial peak, lasting for some 100 ms, a late rebound at about 300 ms, and finished between 500 and 1000 ms after the stimulus. Temperature changes greatly affected the shape of the giant e.p.p., modifying particularly the amplitude and time course of the late rebound.5. The amount of ACh released in response to a single stimulus was measured radiochemically and was found to greatly increase in the presence of 4-AP, explaining the potentiation of the e.p.p. With 4-AP concentrations ranging from 10(-6) M to 10( 4) M, the augmentation of ACh release showed a close correlation with increase of the e.p.p. area.6. The large potentiation of evoked transmitter release occurred in spite of a reduction of ACh stores. After treatment with 10(-4) M-4-AP, the total ACh content was reduced by 30-40% in the absence of any electrical stimulation. The reduction affected to a similar extent the vesicular and extravesicular compartments of ACh. This was accompanied by a general increase in the resting rate of ACh turnover.7. Synaptic vesicles were isolated from small fragments of electric organ, rapidly frozen with our device. Compartmental analysis was carried out by labelling the transmitter pools with a radioactive precursor and it was confirmed that vesicular ACh has a relatively low metabolic rate, whereas free ACh (most probably cytoplasmic ACh) turns over more rapidly. The same finding was obtained after treatment with 4-AP, but the starting levels of ACh and the yield of synaptic vesicles were lower.8. The total ACh content was measured at 30 and 100 ms intervals during the course of the giant e.p.p., and soon after. We found characteristic and significant changes which were (i) an initial fall of total ACh occurring within 100-150 ms, (ii) a transient ACh increase which occurred later and seemed to correspond to the late rebound of the giant e.p.p. and (iii) a steady 20% lowering of total ACh, observed from the end of the giant e.p.p. and lasting for more than 1 s.9. The ATP content of the tissue, during and after the giant e.p.p., followed a time course which was remarkably similar to that of total ACh. A significant ATP/ACh relationship was found in most experiments separately, and in the pooled results with a higher degree of significance.10. Vesicular ACh did not exhibit any significant change during and after the giant e.p.p. Neither the transient initial variations of total ACh nor its later lowering were reflected in similar changes of vesicular ACh. It was therefore the extravesicular pool of ACh which was concerned in the characteristic pattern of changes of total ACh.11. Compartmental analysis of transmitter stores was performed during the course of transmission, after labelling ACh in the tissue with a radioactive precursor. It was found that no detectable transfer of ACh occurred from cytoplasm to vesicles, either during the giant e.p.p., or within the following second.12. The following conclusions were reached. The effect of 4-AP is to cause a very strong and long-lasting potentiation of ACh release, resulting in a giant and complex electrical discharge. Transmitter release under these conditions was not only due to sudden liberation of the preformed, available ACh but also to a marked contribution of new ACh made during the giant e.p.p. These changes in ACh content were very significant and took place exclusively in the extravesicular pool of transmitter. PMID- 6286943 TI - A pharmacological analysis of chloride transport across the amphibian cornea. AB - 1. Active Cl, but not Na, transport across the toad cornea was inhibited by mepacrine, which is a phospholipase A2 inhibitor; trifluoperazine, which blocks the action of calmodulin; and meclofenamic acid, which inhibits synthesis of prostaglandins. Bumetanide and DIDS (4,4'-diisothiocyano-2,2'-stilbene disulphonic acid) have previously been shown to inhibit this Cl transport. The interactions of these antagonists with several agonists that increase Cl transport were studied. 2. The effects of adrenaline, prostaglandin E2, dibutyryl cyclic AMP (DBcAMP) and the Ca ionophore A23187 were inhibited by mepacrine, trifluoperazine and bumetanide. 3. The inhibitory effects of high concentrations of DIDS, however, could be overcome by all of the agonists except DBcAMP. 4. Meclofenamic acid only blocked the effects of A23187. 5. A model is proposed to account for the observed actions and interactions of the various antagonists and agonists on Cl transport. This involves possible roles for Ca, calmodulin and phospholipase A2. PMID- 6286945 TI - Inhibition of the sodium pump by inorganic phosphate in resealed red cell ghosts. AB - 1. Ouabain-sensitive Rb influx was measured into K-free resealed red cell ghosts. The effects of inorganic phosphate (Pi) were examined. 2. Phosphate decreased the magnitude of the influx. Increasing Pi lowered the apparent affinity for both ATP and external rubidium ions. The effects of Pi on the affinity for external Rb were greatest at low ATP concentrations. 3. In the nominal absence of phosphate, increasing ATP from 1 to 100 microM had little effect on the Rb influx from solutions of low (1 microM) rubidium concentrations. In the presence of Pi, ATP increased Rb uptake markedly, even from solutions of low Rb concentration. 4. The above interactions between Pi, ATP and external Rb are consistent with a consecutive scheme for the Na pump in which phosphate is released after potassium binds at the external surface and before ATP binds to release potassium ions to the internal solution. 5. Previous failures to find an effect of phosphate on either the affinity for ATP or that for external potassium (rubidium) ions are shown to be equally consistent with the model. The lack of change of apparent affinity is shown to result from the restricted range of concentrations used in these previous experiments. PMID- 6286944 TI - Reduced ATP concentration as a basis for synaptic transmission failure during hypoxia in the in vitro guinea-pig hippocampus. AB - 1. Experiments were performed to determine whether a decrease in tissue ATP contributes to the rapid failure of cerebral synaptic transmission during hypoxia. Transmission between the perforant path and the dentate granule cells in the in vitro hippocampus was studied.2. Hippocampal slice ATP is decreased by approximately 15% at the time that the evoked response begins to diminish in standard Krebs bicarbonate buffer. This is about 2 min after the onset of hypoxia.3. When transmission failure is accelerated by increasing extracellular K(+) from 4.4 to 13.4 mM, the evoked response begins to decay about 30 sec after exposure to hypoxia. There is no decrease in hippocampal slice ATP at this time.4. However, ATP in the molecular layer (the synaptic region of the tissue) is decreased by approximately 15% at the time the evoked response begins to decay in the slices exposed to elevated K(+) concentration.5. Exposing the hippocampal slice to 25 mM-creatine for 3 hr elevates molecular layer phosphocreatine fourfold. Synaptic transmission during hypoxia survives three times as long as it does in the absence of creatine.6. In the creatine fortified medium, molecular layer ATP no longer declines within 30 sec of hypoxia. However the molecular layer ATP does decline within 90 sec of hypoxia, the time at which the evoked response begins to decay in this creatine-fortified buffer.7. The results establish that ATP in the region of the active synapses is lowered when the first signs of electrophysiological failure appear during hypoxia. They also show that maintaining ATP for longer than normal during hypoxia is associated with a prolonged maintenance of the evoked response. They thus suggest that a decline in ATP is one factor causing hypoxic block of synaptic transmission.8. It is further suggested that the very rapid failure of the electroencephalogram during anoxia may also result from a decline in ATP. PMID- 6286946 TI - Enhancement of synaptic transmission by 4-aminopyridine in hippocampal slices of the rat. AB - 1. The effects of 4-aminopyridine (4-AP) on neurotransmission in hippocampal slices of the rat were studied in vitro. 2. Extracellular recordings of field potentials and intracellular recordings of excitatory and inhibitory post synaptic potentials, action potentials and TTX-resistant spikes were obtained from the somatic and dendritic fields of CA 1 hippocampal neurones. 3. Constant perfusion with, or bolus injection of 4-AP in micromolar concentrations resulted in substantial and persistent enhancement of the amplitude of synaptic field potentials and population spikes with no alteration of the afferent input. Somatic positive waves (p-waves) were increased with local micro-application of 4 AP. 4. Intracellular recordings revealed an increase in the amplitude of e.p.s.p.s. and i.p.s.p.s. I.p.s.p.s were prolonged by 4-AP. 5. The pyramidal cells were, after an initial hyperpolarization, depolarized by up to 10 mV during exposure to 4-AP; the membrane conductance was not consistently changed. 6. Spontaneous shifts of the membrane potential were described as giant e.p.s.p.s and i.p.s.p.s. Spontaneous i.p.s.p.s were increased in frequency and amplitude. 7. Paired-pulse facilitation of e.p.s.p.s was reduced by 4-AP suggesting a presynaptic modulation of transmitter output. 8. It is concluded that 4-AP enhances transmitter release at both excitatory and inhibitory synapses in the hippocampus. PMID- 6286947 TI - Reinnervation of denervated parasympathetic neurones in cardiac ganglia from Rana pipiens. AB - 1. The sequence of events during reinnervation of the cardiac ganglion in the frog following interruption of the vagosympathetic nerve supply was studied with both electrophysiological and morphological techniques. 2. When cardiac ganglia were denervated by crushing the vagosympathetic nerve supply to the heart all synaptic endings on parasympathetic ganglion cells degenerated. Vacated post synaptic densities were detected on denervated neurones for periods of at least 7 weeks. 3. The earliest signs of reinnervation were subthreshold responses evoked by stimulating the regenerating vagosympathetic trunks 2 1/2-3 weeks after crushing the cardiac branches of the vagus nerves. Analysis of the reversal potentials of these responses indicated that these synapses were distant from the cell body. 4. At slightly longer times (4-5 weeks), regenerating synapses could be recognized on post-ganglionic axons; no synapses were detected on the neuronal perikarya at these times. 5. By 6-7 weeks following denervation, vagal synapses reinnervated neuronal perikarya as well as post-ganglionic axons. At the same time, vacated post-synaptic densities declined in number. Furthermore, vagal stimulation at this stage evoked large, suprathreshold post-synaptic potentials. 6. These studies indicate that post-ganglionic axons are the initial sites for reinnervation of parasympathetic neurones in the heart. Only some time later are neuronal perikarya reinnervated and ganglionic transmission completely restored. PMID- 6286949 TI - Amiloride-sensitive sodium channels in rabbit and guinea-pig gall-bladder. AB - 1. The effects of low amiloride concentrations (less than 2.5 x 10(-5) M) on the apical membrane (Vm) and transepithelial (Vms) potential differences, transepithelial resistance (Rep) and apical/basolateral membrane resistance ratio (Rm/Rs) have been determined in the gall-bladders of the rabbit and guinea-pig. 2. Vm was hyperpolarized, Vms decreased towards negative values, Rm/Rs increased, but Rep remained unchanged. 3. K+ selectivity of the apical membrane was also checked by increasing luminal K+ concentration from 5.9 to 29.4 mM; the corresponding change in the apical electromotive force was much higher in the rabbit than in the guinea-pig. 4. Pre-incubations with Cl--and HCO3--free salines did not modify K+ selectivity in the rabbit, but nearly doubled it in the guinea pig. 5. Pre-incubations with control solutions containing 10(-5) M-amiloride in the lumen converted the apical membrane of rabbit gall-bladder into a perfectly K+-perm-selective membrane; the same results were obtained in the guinea-pig only on bathing the tissue with Cl-- and HCO3--free salines together with 10(-5) M amiloride. 6. It is suggested that in the apical membrane of rabbit gall-bladder K+ and Na+ conductances exist, whereas in guinea-pig gall-bladder both K+, Na+ and anion (Cl- and/or HCO3-) conductances are present; in both species conductive Na+ pathways are inhibited by amiloride. . PMID- 6286950 TI - Mechanisms of relaxation induced by activation of beta-adrenoceptors in smooth muscle cells of the guinea-pig mesenteric artery. AB - Relaxation of smooth muscle cells induced by activation of beta-adrenoceptors was investigated in intact and skinned muscles of the guinea-pig mesenteric artery.1. In concentrations over 10(-7) M, isoprenaline reduced the resting tone of intact preparations and also the amplitude of K contractions. When Ca was applied after previous superfusion with Ca-free solution, the amount of Ca accumulated into storage sites was increased by isoprenaline in polarized and depolarized ([K](o) 128 mM) muscles. The amount of Ca stored increased even further when procaine and isoprenaline were applied simultaneously during store loading.2. Isoprenaline increased the concentration of cyclic AMP as determined by radioimmunoassay. Application of isoprenaline at a concentration of 10(-7) M increased cyclic AMP from 2.2+/-0.3 to 2.8+/-0.6 p-mole/mg wet weight and at 10(-6) M increased it to 4.5+/-0.8 p-mole/mg wet weight after 5 min incubation (n = 4).3. Application of cyclic AMP (3 x 10(-6) M) with cyclic AMP-dependent protein kinase (50 mug/ml.) had no effect on the pCa-tension relationship in the skinned muscles. However, an increased concentration of cyclic AMP (> 10(-5) M) suppressed the Ca-induced concentration only in the presence of protein kinase. This protein kinase (50 mug/ml.) alone had no effect on the Ca-induced contraction.4. In skinned fibres, the Ca store could be loaded by applying low concentrations of Ca. If cyclic AMP (3 x 10(-6) M) with protein kinase (50 mug/ml.) was applied during the loading procedure, the amount of Ca accumulated by the store increased if the loading solution contained 10(-6) M-Ca applied for 2 min or less, but if the loading solution was applied for 3 min, or if higher Ca concentrations were used, the presence of cyclic AMP with protein kinase decreased the store size, suggesting that a Ca-induced Ca-release mechanism was also being activated.5. In skinned muscles, accumulation of Ca into the store site in the presence of cyclic AMP (3 x 10(-6) M) with protein kinase (50 mug/ml.) was further accelerated by simultaneous applications of procaine (5 mM), as here the Ca-induced Ca-release mechanism was suppressed.6. These results indicate that activation of beta adrenoceptors by isoprenaline increases the amount of cyclic AMP in the intact muscles, and leads to an increase in Ca accumulation into the store site. In the skinned muscles, the Ca-induced Ca-release mechanism is activated by cyclic AMP and the Ca receptor for contraction (leiotonin C or calmodulin) is somewhat suppressed. These effects of exogenously applied cyclic AMP require the presence of protein kinase. The relaxation following beta-adrenoceptor activation is more likely to involve Ca extrusion from the cell and accumulation of Ca in internal storage sites than suppression of the binding of calmodulin with the myosin light chain kinase. PMID- 6286951 TI - Effects of goniopora toxin on non-adrenergic, non-cholinergic response and purine nucleotide release in guinea-pig taenia coli. AB - 1. Effects of goniopora toxin (GPT) on non-adrenergic, non-cholinergic inhibitory responses were examined in isolated guinea-pig taenia coli and the transmission mechanism was analysed.2. GPT (20-50 nM) gradually augmented the relaxation and rebound contraction induced by electrical transmural stimulation in the presence of guanethidine and atropine. These effects were abolished by tetrodotoxin.3. The resting tension and the response to exogenously applied ATP were little affected by GPT.4. The action potentials and the electrotonic potentials recorded by a double sucrose-gap method were little affected by GPT.5. These results indicate that the augmentation is elicited by a direct action of GPT on the non adrenergic, non-cholinergic neurones, rather than on the smooth muscles per se.6. GPT augmented the amplitude of the inhibitory junction potential (i.j.p.) recorded in the presence of guanethidine and atropine. GPT also enhanced the (3)H efflux in response to electrical transmural stimulation of preparations pre loaded with [(3)H] adenosine. Tetrodotoxin markedly inhibited or abolished the i.j.p. and (3)H efflux.7. These results would suggest that augmentation of the non-adrenergic, non-cholinergic responses is due to increased transmitter release from the nerve. Under conditions in which the transmitter release was augmented, the non-adrenergic, non-cholinergic response showed an essentially similar pattern to that obtained under normal conditions; therefore the inhibitory response is probably due to a putative transmitter, possibly ATP or a related nucleotide. PMID- 6286948 TI - Slowing effects of dopamine and calcium-channel blockers on frequency of sodium spikes in rat pars intermedia cells. AB - 1. Spontaneous discharge of action potentials (Na spikes) in cells isolated from rat pars intermedia was slowed or arrested by Co(2+), Ni(2+) or Mn(2+), which block voltage-dependent Ca channels in these cells. The amplitude of persisting spikes was undiminished. The effects resembled those of dopamine.2. Action potential frequency decreased when the Ca(2+) concentration was lowered to 0.1 mM and increased when the Ca(2+) concentration was raised from this level to 1 mM or 2 mM or when Ba(2+) (2 mM) was introduced. These effects, together with those of Co(2+), Ni(2+) and Mn(2+), are consistent with the possibility that Ca(2+) participates in the regulation of spike discharge.3. Verapamil, methoxyverapamil (D600), and nifedipine reduced the amplitude of the individual Na spikes in concentrations that had little effect on voltage-dependent Ca channels. Action potential frequency was comparatively little affected by these drugs.4. K(+) (15 mM) stimulated action potential frequency and this effect too was suppressed by dopamine or Co(2+).5. The effect which dopamine had of slowing spontaneous discharge, like the inhibitory effect on secretion, was blocked by metoclopramide. But otherwise the mechanism is unclear: dopamine blocked voltage dependent Ca channels in some cells but not in most others.6. The effects of K(+) and Ba(2+) of eliciting spikes, the suppression of Na-spike discharge by Co(2+) and related Ca-channel blocking cations, and the unspecific effects of the organic ;Ca channel blockers', all have implications for the use of these substances as tools to analyse stimulus-secretion coupling. PMID- 6286953 TI - Ions and excitable membrane. 5th GIF Lectures in Neurobiology. PMID- 6286954 TI - Transmembrane electrical potential of excitable membranes: a pore analysis influence of surface charges and surface dipoles. AB - A treatment is proposed in order to establish the general expression of the zero current transmembrane potential of excitable membranes. The membrane model considered here is that of a hydrocarbon layer which is impermeable to ions and which represents the lipid bilayer matrix. In this matrix are incorporated ionic channels. The ion transport process through the channels is described by the absolute-rate theory applied to pores which are seen as chains of potential energy maxima and minima. Only one of the energy barriers corresponds to the gate step, and it is strongly dependent on the transmembrane potential. The kinetic equation is related to the zero current, to electrostatic boundary conditions and to the Gouy-Chapman equation for the aqueous diffuse layer. PMID- 6286952 TI - Properties of miniature excitatory junctional currents at the locust nerve-muscle junction. AB - 1. Miniature excitatory junctional currents (m.e.j.c.s) were examined in conditions where inward current was carried mainly by Na(+) (i.e. in normal medium, Ca(2+)-free medium and Cl(-)-free medium). M.e.j.c.s were also examined in isotonic Ca(2+) where the inward post-synaptic current was carried mainly by Ca(2+).2. In normal medium, mean m.e.j.c. amplitude = 2.34+/-0.05 nA. The decay time constant of m.e.j.c.s (excluding a small percentage with abnormal shapes) was tau(m.e.j.c.) = 2.62+/-0.11 msec (V(m) = -80 mV, T = 22 degrees C). Decay time was not markedly changed in Ca(2+)-free or Cl(-)-free medium. tau(m.e.j.c.) approaches the life-time of glutamate activated junctional channels.3. Excitatory junctional currents, evoked by nerve impulses, decayed slightly faster than m.e.j.c.s obtained in the same fibres. Extracellularly recorded m.e.j.c.s and voltage-clamped m.e.j.c.s were similar in time course.4. tau(m.e.j.c.) decreased exponentially with membrane hyperpolarization. An e-fold change was produced by 182.+/-24.8 mV change in V(m).5. The dependence of mean m.e.j.c. amplitude on clamp potential showed a slight non-linearity at hyperpolarized levels. The equilibrium potential for transmitter action was close to 0 mV in normal solution as well as in Ca(2+)-free and Cl(-)-free solutions.6. The kinetics of junctional channels are altered in isotonic Ca(2+). M.e.j.c. amplitude was reduced to about one-third normal size; mean m.e.j.c. = 0.74+/-0.03 nA. The decay time becomes markedly briefer, tau(m.e.j.c.) = 1.01+/-0.08 msec, indicating a reduction in mean channel life-time (V(m) = -80 mV, T = 22 degrees C).7. A population of slow time course and composite m.e.j.c.s appear when muscle fibres are hyperpolarized in isotonic Ca(2+), thus producing a prolongation in mean tau(m.e.j.c.). This results from an influence of post-synaptic membrane potential on presynaptic transmitter release. If such m.e.j.c.s are ignored the voltage dependence of tau(m.e.j.c.) of the remaining events is abolished or even reversed indicating that voltage sensitivity of channel life-time is altered in isotonic Ca(2+). The equilibrium potential for transmitter action may be slightly more positive than normal.8. We estimate that a single packet of neurally released transmitter normally opens, on average, 250 ion channels at these junctions. PMID- 6286955 TI - Voltage modulation of Na+/K+ transport in human erythrocytes. AB - For a cell to perform certain functions it presumably must maintain a steady state transmembrane potential. This potential differential can take several forms, e.g. a proton gradient, an ionic concentration gradient, or an asymmetric distribution of charges in membraneous proteins and lipids. We report here a study which indicates that an externally applied electric field can induce opening/closing of membrane channels, including Na+/K+ ATPase channels. The method utilizes a continuous AC stimulation of cell suspensions. Reversible channel opening/closing of human erythrocytes in an isotonic suspension can be demonstrated by directly measuring the conductivity of the bulk suspension. Channel opening occurs at a field intensity of 10 V/cm, which corresponds to a maximum transmembrane potential of about 6 mV, when the frequency of AC field is maintained below 1 kHZ. The apparent half-time of the channel opening decreases with an increase in the stimulating voltage, and reaches a plateau value of 2 sec beyond 24 V/cm (15 mV of transmembrane potential). When the AC field is removed, these channels close with a half-time of 10.2 sec. Of the channels opened, roughly 20% belongs to the Na+/K+ ATPase, as this fraction of conductance signals can be quantitatively blocked by a specific inhibitor ouabain in a low ionic medium. The AC field appears to stimulate active Na+/K+ transport against a concentration gradient in erythrocytes. At a field strength of 24 V/cm, the net transport against a controlled sample is about 1 mM 42K+ ion per hour under our experimental condition. This translates into a rate of 60 K+ ions per channel per second. The AC field also increases the K+/Na+ ratio of cytoplasmic concentration in the stimulated sample, indicating an active extrusion of Na+ ion from the cells. Higher consumption of ATP is not detected in the stimulated cells as compared with unstimulated cells. As expected, an addition of ouabain in the external medium abolishes the above effects. Experiments described here would demonstrate that the phenomena observed are due to the effect of the field induced transmembrane potential. The significance of these observations are discussed in the light of the recent discovery that transmembrane potential is an important driving force for certain cellular functions. PMID- 6286956 TI - Ion flow gating by the acetylcholine system: kinetics of isolated receptor and esterase and of receptor-mediated ion flux. AB - 1. Basic features of the elementary bioelectric signals such as miniature endplate-potentials are molecularly interpreted on the basis of relaxation kinetic data of isolated acetylcholine receptor and acetylcholinesterase. Electrophysiological and molecular data suggest an essentially sequential processing of acetylcholine by receptor and esterase. 2. Flux measurements with sealed biomembrane fragments containing acetylcholine receptor show that the ion transporting conformation of the receptor-channel is a short-lived metastable state. In the presence of neuroactivators the receptors inactivate. The description of the flux-inactivation requires a cyclic reaction scheme similar to the desensitization scheme of KATZ and THESLEFF (1957). 3. The recently introduced concept of integrated flux rate coefficients permits us to derive gating mechanisms from flux data under well-defined experimental conditions: sealed biomembrane vesicles, activator concentration, type of transported ion. 4. With respect to activation and inactivation and the metastability of the ion conducting conformation, there are fundamental similarities between the axonal Na+ ion channel and the acetylcholine receptor-channel. PMID- 6286957 TI - Ionic mechanism underlying muscarinic acetylcholine response in the rabbit sinoatrial node. AB - 1. The clusters composed of 10-20 cells of the rabbit sinoatrial node were prepared by coronary perfusion of collagenase and their response to ionophoretic application of ACh was recorded. The hyperpolarizing response showed a sigmoidal onset without any clear latency when the ACh pipette was positioned close to the cell surface. 2. Equations describing the kinetics and amplitude of the ACh induced K current were determined based on the reported experimental results obtained by relaxation and noise analysis. 3. The kinetics of the ACh-induced K current simulated well the time course of the ACh response and also the effect on the spontaneous action potential when data on the kinetics was incorporated into the S-A node pacemaker mathematical model. PMID- 6286958 TI - Reconstitution of the sodium channel with partially solubilized lobster nerve membrane. AB - Reconstitution experiments were carried out with particles obtained from lobster nerve plasma membrane preparations by detergent treatment, differential centrifugation and ammonium sulfate fractionation. The NA channel activity of the three fractions obtained, which have different amounts of the same peptides present in the original membrane, appears related to their content in a large component which does not enter the 9% polyacrylamide gel and in peptides with 220,000 and 110,000 apparent molecular weight. Other reconstitution experiments made with two fractions obtained by detergent treatment, differential centrifugation and gel exclusion chromatography, revealed that the Na channel active fraction contains the material which does not enter the gel in addition to the 220,000 and 110,000 molecular weight peptides. The other fraction was inactive and does not contain those components. The 220,000 dalton peptide has a molecular weight similar to those determined for the tetrodotoxin-saxitoxin receptor and the scorpion toxin receptor of the Na channel. Whether any of the other peptides is a Na channel constituent is unknown at present. PMID- 6286959 TI - Inactivation of the Na permeability in squid giant axons. AB - Inactivation of the Na permeability has been studied in intact and perfused squid giant axons with the voltage clamp method. The main results are: 1. Upon depolarization inactivation develops along an exponential time course; the upper limit for an initial delay in the development of inactivation is 50-100 musec. 2. Adding 20-40 mM KCl to K-free external solution accelerates the development of inactivation and slows its removal. 3. Scorpion venoms increase the maintained conductance, i.e. make inactivation less complete; the voltage dependence of the maintained conductance is different from that of the peak conductance. PMID- 6286960 TI - Modulation of nerve membrane sodium channels by chemicals. AB - 1. Modulations of sodium channel kinetics by grayanotoxins and pyrethroids have been studied using voltage clamped, internally perfused giant axons from crayfish and squid. 2. Grayanotoxin I and alpha-dihydrograyanotoxin II greatly depolarize the nerve membrane through an increase in resting sodium channel permeability to sodium ions. 3. Grayanotoxins modify a fraction of sodium channel population to give rise to a slow conductance increase with little or no inactivation, and the slow conductance-membrane potential curve is shifted toward hyperpolarization. This accounts for the depolarization. 4. The tail current associated with step repolarization during the slow current in grayanotoxins decays with a dual exponential time course. 5. (+)-trans tetramethrin and (+)-trans allethrin also modify a fraction of sodium channel population in generating a slow current, which attains a maximum slowly and decays very slowly during a maintained depolarizing step. The membrane is depolarized only slightly. 6. The tail current associated with step repolarization during the slow current in the pyrethroids is very large in initial amplitude and decays very slowly. 7. The rate at which the sodium channel arrives at the modified open state in the presence of pyrethroids is expressed by a dual exponential function, and the slow phase disappears following removal of the sodium inactivation mechanism by internal perfusion of pronase. 8. A kinetic model is proposed to account for the actions of both grayanotoxins and pyrethroids on sodium channels. Both chemicals interact with the channel at both open and closed states to yield a modified open state which results in a slow sodium current. PMID- 6286961 TI - The rise and fall of electrical excitability in the oocyte of Xenopus laevis. AB - 1. An electrically excited (gated) sodium selective channel has been found in the Xenopus laevis oocyte, a cell membrane previously considered non-excitable. 2. The channel is produced by prolonged depolarization of the membrane and is removed by prolonged repolarization. Both processes are very dependent on temperature and potential. 3. Once produced, the channel can be opened and closed electrically, but does not show inactivation as is found in other sodium selective channels. 4. The sodium selectivity and the electrical gating properties of this channel make it a potentially useful candidate for the study of these general channel characteristics. The fact that this membrane can be made to show these properties reversibly offers the possibility of the studying the origins of this channels. PMID- 6286962 TI - Pharmacological characterization of ionic channels in unmyelinated axons. PMID- 6286963 TI - Properties of the slow K+ current of the nodal membrane. AB - 1. In voltage clamped myelinated nerve fibres, some properties of the slow K+ current (Dubois, 1981 b) were analysed under different experimental conditions. 2. The instantaneous slow current-voltage relationships are linear in low and high K+ solutions, and the variation of the reversal potential with external K+ concentration agrees with the predicted Nernst changes for the K+ equilibrium potential. 3. Li+, Na+ and Cs+ are virtually impermeant in the slow channels. Rb+ is permeant and the permeability ratio PRb/PK (calculated from the reversal potential and the Goldman-Hodgkin-Katz equation) is a function of the external K+ and Rb+ concentrations. 4. External and internal Cs+ block respectively the slow inward and outward K+ current and the block is voltage dependent. 5. The closing kinetics of the slow channels are independent of the external K+ concentration but depend on the conditioning pulse duration. PMID- 6286964 TI - Differences in electrophysiological properties of motor and sensory nerve fibres. AB - Differences in the electrophysiological properties of amphibian motor and sensory nerve fibres are reviewed. It is concluded that the differences are mainly due to an altered K-conduction system which suggests the presence of different K channels in the two types of nerve fibres. PMID- 6286965 TI - Ca entry upon depolarization of nerve. AB - The data reviewed and the experiments presented show that a steady depolarization of a squid giant axon membrane leads to a large but transient Ca entry that is virtually totally dependent upon [Na]i. Since conventional membrane channels do not behave in this way whereas a countertransport reaction is defined as one requiring the participation of ions on opposite sides of the membrane, it is considered reasonable to identify the Ca entry that has been measured as a Na/Ca exchange. Since the enhanced Ca entry is initiated by a reduction in membrane potential, a further conclusion is that the Na/Ca exchange reaction involves a non-electroneutral exchange of changes. The physiological importance of these findings of a Nai-dependent Ca influx in nerve may be that it assures larger Ca entries following rapid tetanic stimulation (because of the increase in Nai) and this may be necessary to produce a Cai signal that is detectable over the increased Cai background as a result of the tetanic stimulation. In smooth and cardiac muscle, increases in Nai surely play a regulatory role in increasing Ca entry with depolarization and hence in increasing tension. PMID- 6286967 TI - Primary malignant fibrous histiocytoma of bone. PMID- 6286966 TI - Ionic channels of Paramecium: from genetics and electrophysiology to biochemistry. AB - This paper reviews the combined genetical, electrophysiological and biochemical analysis of excitation that has been carried out in Paramecium. Paramecium cells display graded Ca++ action potentials in response to a variety of stimuli. These action potentials regulate the orientation of the ciliary beat hence the cell's swimming behaviour. A large array of mutants displaying altered behaviour have been isolated and mapped to over 20 loci. Detailed electrophysiological analyses have been carried out on several classes of mutants revealing defects in specific ion channels in some cases. Mutants have proven very useful to analyze channel properties, to unravel interactions between channels and to discover the function of these channels in a variety of cellular processes. Some important channels are located in the ciliary membranes and cilia as well as ciliary membranes can now be purified in high purity and reasonable yield. These fractions have been used recently in a variety of biochemical approaches to gain insight into the molecular components of the excitation machinery. Specific alterations in some minor membrane proteins have been found in two mutants as well as a specific defect in sphingolipids in a third mutant. Those alterations had to be distinguished from large scale variations in membrane proteins and lipids that occur in this organism in response to modifications in growth conditions. Several other recent biochemical developments are reviewed and the advantages as well as the difficulties of the genetic approach to the molecular study of biological processes are discussed. PMID- 6286968 TI - [The liver masses in ultrasonography. Report on 400 cases (author's transl)]. AB - On the basis of 400 cases of liver masses, the authors have drawn conclusions that can be used to orient etiological investigations: --one or more well demarcated transsonic formations always correspond to a cystic etiology: --a hyperechoic metastatic liver points towards a primary cancer of the digestive tract whereas a hypoechoic metastatic liver is only rarely caused by such a cancer. The authors also propose a strategy for the exploration and surveillance of isolated hyperechoic or transsonic nodules of no more than 3 cm. PMID- 6286969 TI - Oxidation of glutathione by the myeloperoxidase system. AB - Oxidation of glutathione (GSH) by the myeloperoxidase (MPO) system was studied. The combination of MPO, H2O2, and a halide ion oxidized GSH. This occurred at a H2O2 concentration too low to oxidize GSH by itself. The MPO-mediated oxidation of GSH required the simultaneous presence of MPO, H2O2, and a halide ion. The system had an acid pH optimum of pH 5.5-6.0. Iodide was more effective than bromide which in turn was more effective than chloride. The oxidative product was shown to be GSSG, since it could be reduced back to GSH by glutathione reductase and NADPH. The MPO-mediated oxidation of GSH may be one mechanism by which this system damages microorganisms. PMID- 6286970 TI - Species- or isozyme-specific enzyme inhibitors. 7. Selective effects in inhibitions of rat adenylate kinase isozymes by adenosine 5'-phosphate derivatives. AB - Monosubstituted derivatives of adenosine 5'-phosphate (AMP) with substituents of 1-3 atoms or group replacements at any of 11 positions have been synthesized and examined as substrates and inhibitors of the rat muscle adenylate kinase isozyme (AK-M), and the rat AK II and III isozymes predominant in poorly differentiated hepatoma tissue and normal liver tissue, respectively. Inhibition indexes of the compounds were expressed as KM (AMP)/Ki for competitive inhibition or as KM (AMP)/KM when only KM was available. Substituents at N(1), N6, or C(8) or on ionizable phosphate oxygen reduced inhibition below measurable levels; 2'-deoxy AMP and adenosine 5'-sulfate had identical inhibition indexes with all three isozymes; compounds with substituents at C(2), O(2'), O(3'), C(4'), C(5'), or O(5') had higher inhibition indexes with AK-M than with AK II or III and the same or similar indexes for AK II and III. The most effective and/or selective inhibitors were 2-NHMe-AMP (index with AK-M, 0.2; index ratio, AK-M/AK III, 9.1), 2'-O-Me-AMP (index with AK-M, 0.14; index ratio, AK-M/AK III, 8.2), 2',3'-O-CMe2 AMP (index with AK-M, 0.25; index ratio, AK-M/AK II, 6.6), 4'-allyl-AMP (index with AK-M, 0.97; index ratio, AK-M/AK III, 8.1), and 5'(S)-Et-AMP (index with AK M, 0.64; index ratio, AK-M/AK II, 11.2). The study provides additional evidence that the attachment of simple substituents to various atoms in turn of a substrate is a potentially useful approach in early stages of the attempted design of isozyme-selective inhibitors. PMID- 6286972 TI - Leser-Trelat sign in mother and daughter with breast cancer. AB - The Leser-Trelat sign is the sudden appearance and rapid increase in size and number of seborrhoeic keratoses in association with cancer. Twenty cases of this unusual phenomenon have so far been reported in the world literature. More than half involve adenocarcinomas and none was shown to be familial. We report a unique example of a 41-year-old black female and her 74-year-old black mother, both of whom have demonstrated classical clinical-pathological evidence of the Leser-Trelat sign and adenocarcinoma of the breast. The remainder of the family history was negative for cutaneous lesions and cancer. The aetiological and pathogenetic significance of the Leser-Trelat sign in association with carcinoma of the breast in this mother and daughter remains obscure. PMID- 6286971 TI - Isozyme patterns and protein profiles in neuromuscular disorders. AB - The isozyme patterns of six different enzymes and the polypeptide profiles of soluble proteins have been examined in muscle biopsy specimens from 74 patients with a wide variety of neuromuscular disorders. About half of the samples showed unusual features in at least one, and often several, of the enzymes and proteins tested. The extent of the biochemical abnormalities was roughly proportional to the severity of the disorders. In all cases the unusual isozymes and polypeptide profiles seemed to reflect a reversion to the fetal pattern of gene expression. However, this change appeared to occur in extant muscle and was not dependent on the appearance of new muscle fibres. Among the enzymes, phosphoglycerate mutase followed by creatine kinase appeared to be the most sensitive index of muscle disorder. The extent of the change in the muscle creatine kinase isozyme pattern was not correlated with the levels of serum creatine kinase activity. PMID- 6286973 TI - Active and passive Na+ fluxes across the basolateral membrane of rabbit urinary bladder. AB - The apical membrane of rabbit urinary bladder can be functionally removed by application of nystatin at high concentration if the mucosal surface of the tissue is bathed in a saline which mimics intracellular ion concentrations. Under these conditions, the tissue is as far as the movement of univalent ions no more than a sheet of basolateral membrane with some tight junctional membrane in parallel. In this manner the Na+ concentration at the inner surface of the basolateral membrane can be varied by altering the concentration in the mucosal bulk solution. When this was done both mucosal-to-serosal 22Na flux and net change in basolateral current were measured. The flux and the current could be further divided into the components of each that were either blocked by ouabain or insensitive to ouabain. Ouabain-insensitive mucosal-to-serosal Na+ flux was a linear function of mucosal Na+ concentration. Ouabain-sensitive Na+ flux and ouabain-sensitive, Na+-induced current both display a saturating relationship which cannot be accounted for by the presence of unstirred layers. If the interaction of Na+ with the basolateral transport process is assumed to involve the interaction of some number of Na+ ions, n, with a maximal flux, MMAX, then the data can be fit by assuming 3.2 equivalent sites for interaction and a value for MMAX of 287.8 pM cm-2 sec-1 with an intracellular Na concentration of 2.0 mM Na+ at half-maximal saturation. By comparing these values with the ouabain sensitive, Na+-induced current, we calculate a Na+ to K+ coupling ratio of 1.40 +/- 0.07 for the transport process. PMID- 6286974 TI - Triphenylmethylphosphonium cation distribution as a measure of hormone-induced alterations in white adipocyte membrane potential. AB - Triphenylmethylphosphonium (TPMP+) partitions into the mitochondrial and cytosolic compartments in the rat white adipocyte in a potential-dependent fashion. The relationship between [3H]TPMP+ distribution, intracellular cAMP generation and lipolysis in response to hormones and cAMP-mimetic compounds was examined. Half-maximal [3H]TPMP+ efflux and glycerol release were produced by 15 and 9 nM adrenocorticotropin, 170 and 110 nM 1-epinephrine, 70 and 27 microM isobutylmethylxanthine and 800 and 750 microM dibutyryl cAMP, respectively. Hormone-stimulated cAMP generation was also correlated with [3H]TPMP+ efflux and lipolysis in terms of concentration dependency. In kinetic experiments, glycerol release and [3H]TPMP+ efflux in response to adrenocorticotropin or cholera toxin proceeded over a similar time course, whereas an earlier rise in cAMP generation was detected. The depolarizing effect of lipolytic compounds was localized to the mitochondrial compartment. When cells were incubated in elevated-[K+]0 buffer, the stimulatory effect of dibutyryl cAMP on [3H]TPMP+ efflux and lipolysis persisted, suggesting that maintenance of the plasma membrane potential is not critical for demonstration of these responses. When the extracellular concentration of serum albumin, which provides binding sites for free fatty acids, was increased from 1 to 3%, an increase in glycerol release and a decrease in [3H]TPMP+ efflux was observed. We suggest that intracellular free fatty acid accumulation in response to lipolytic agents causes dissipation of the mitochondrial membrane potential and efflux of [3H]TPMP+ from the organelle and cell. PMID- 6286975 TI - Properties of sodium and potassium channels of the squid giant axon far below 0 degrees C. AB - Squid giant axon could be excited in concentrated glycerol solutions containing normal concentrations of electrolytes, when osmolalities of solutions inside and outside the axon were matched. These glycerol solutions did not freeze at the temperature as low as -19 degrees C. The nerve excitation in these solutions were observed at this low temperature. The excitation process at this low temperature was slowed down and time constants of the excitation kinetics were several hundredfold larger than those in normal seawater at 10 degrees C, under which temperature the squid habituated. The temperature coefficients for the electrophysiological membrane parameters under this condition were larger than those in normal seawater above 0 degrees C. The Q10 value for the conduction velocity was 2.0 and that of the duration of the action potential was around 8.5. The time course of the membrane currents was also slowed with the Q10 value of around 5 and the magnitude decreased with the Q10 value of around 2 as the temperature was lowered. The Q10 values for the kinetics of the on process of the Na-channel were around 4.5 and were almost the same as those of the off process of the Na-channel in the wide range of the temperature below 0 degrees C. The Q10 value of the on process of K-channel was around 6.5 and was larger than those for Na-channel. The Q10 values increased gradually as the temperature was lowered. PMID- 6286976 TI - Dopaminergic inhibition of vasopressin-stimulated water flow in the toad bladder: evidence for local formation of dopamine. PMID- 6286978 TI - Initiation of sequential packaging of bacteriophage P22 DNA. PMID- 6286977 TI - Novel mutations of Escherichia coli that produce recombinogenic lesions in DNA. V. Recombinogenic plasmids from arl mutants of Escherichia coli are unusually sensitive to nuclease S1 and partially deficient in cytosine methylation at C-C (A/T)-G-G sequences. PMID- 6286979 TI - Secondary structure in ribonuclease. I. Equilibrium folding transitions seen by amide circular dichroism. PMID- 6286980 TI - Secondary structure in ribonuclease. II. Relations between folding kinetics and secondary structure elements. PMID- 6286982 TI - Histological and enzyme histochemical studies on the nephrons of the freshwater fishes, Cyprinus carpio and Carassius auratus. AB - The nephrons of carp (Cyprinus carpio) and goldfish (Carassius auratus) were examined histologically and also histochemically for enzymes. In both species the distal and collecting tubules have much wider lumens than do the other renal tubules; thus urine probably flows more slowly in these larger tubules. Enzyme histochemistry shows that epithelium of the neck and proximal and intermediate tubules respires anaerobically, whereas that of the distal and collecting tubules respires aerobically. The distribution of Na-K-ATPase in the distal and collecting tubules indicates that they also transport sodium actively. The slow flow of urine and the energy produced by aerobic metabolism probably increase the efficiency of active transport. PMID- 6286981 TI - HineI is an isoschizomer of HinfIII restriction endonuclease. PMID- 6286983 TI - Increase in myocardial cell cGMP concentration in pressure-induced myocardial hypertrophy. PMID- 6286984 TI - The interaction of calcium antagonists (slow channel blockers) with myocardial alpha adrenoceptors. PMID- 6286985 TI - Effect of mineral oil and/or cholestyramine in the diet on biliary and intestinal elimination of 2,4,5,2',4',5'-hexabromobiphenyl in the rhesus monkey. AB - Addition of mineral oil to the diet (5%) of two rhesus monkeys that were dosed 29 wk earlier with 2,4,5,2',4',5'-hexabromobiphenyl (HBB) produced a 175% increase in fecal excretion of HBB. A third rhesus monkey was provided with a complete biliary bypass, which permitted the reintroduction of the monkey's own bile or the introduction of the same amount of exogenous bile, which was obtained from control donor monkeys. This experimental design made it possible to measure the portion of fecal excretion that was due to biliary elimination and the portion that was due to intestinal elimination. The effect of mineral oil and/or cholestyramine (CSA) on biliary and intestinal elimination of HBB in the rhesus monkey was then investigated. The results show that (1) fecal excretion of HBB and/or metabolites is due to both biliary and intestinal elimination; (2) during the first 2 wk after completion of dosing, mineral oil does not influence fecal excretion of HBB significantly; (3) 6-7 wk after completion of dosing, mineral oil enhances fecal excretion of HBB by 50%; (4) 8 wk after the last dose of HBB, 4% CSA in the diet increases fecal excretion of HBB and/or metabolites by about 50%; (5) mineral oil specifically stimulates intestinal elimination of HBB; and (6) combined administration of mineral oil and CSA results in an additive effect. PMID- 6286987 TI - Surgery for tumors of the deep lobe of the parotid gland. PMID- 6286986 TI - Effect of the insecticides toxaphene and carbaryl on induction of lung tumors by benzo[a]pyrene in the mouse. AB - The insecticides toxaphene and carbaryl, when fed in the diet alone for 20 wk, were not tumorigenic to female A/J mice. Dietary levels of these insecticides were investigated for their effects on the incidence of lung tumors induced by oral administration of benzo[a]pyrene (BP). A significant reduction in BP-induced lung tumors was found after feeding 100 ppm toxaphene for 12 wk or 200 ppm for 20 wk. In contrast, 1000 ppm carbaryl fed for 20 wk caused a significant enhancement of BP-induced lung tumors. Mice that received toxaphene in the diet alone, or toxaphene and BP, showed an increase in BP hydroxylase activity in the liver and a decrease in enzyme activity in the lung. Carbaryl and BP increased BP hydroxylase activity in the lung without altering enzyme activity in the liver. Inhibition of lung BP hydroxylase activity was paralleled by a reduction in BP induced lung tumors in mice fed toxaphene. Conversely, increased lung BP hydroxylase activity was associated with an enhancement of BP-induced lung tumors in animals fed carbaryl. The metabolism of BP by organs susceptible to BP-induced tumors and possible mechanisms for interactions with the insecticides are discussed. PMID- 6286988 TI - Pharmacokinetics of polymyxin B administered via the bovine mammary gland. AB - Polymyxin B was infused into normal, chronically inflamed, and acutely inflamed quarters of the mammary gland of lactating cows at dosages ranging between 1 and 2 million units (100-200 mg) per quarter. Samples of milk from treated and non treated quarters, jugular venous blood, subcutaneous abdominal (mammary) venous blood, and urine were collected at intervals after treatment and were assayed using microbiological test methods for polymyxin B concentrations. The drug was not absorbed from normal and chronically inflamed quarters; more than 90% of the infused dose was recovered in milk within 24 h after treatment, and drug residues were detected up to the ninth milking. Drug concentrations in milk from acutely inflamed quarters were significantly lower than in milk from normal quarters; 55% of the infused dose was recovered in the milk within 24 h after treatment. The drug was detected in milk from non-treated quarters, in blood from the subcutaneous abdominal vein, and in the urine during 36-48 h after acutely inflamed quarters were infused with the drug. These data indicate that polymyxin B is well distributed throughout, and is absorbed to a significant degree into the systemic circulation from the acutely inflamed udder. PMID- 6286989 TI - Unique features of retrovirus expression in F/St mice. AB - F/St mice are unique in producing high levels of both ecotropic and xenotropic murine leukemia virus. The high ecotropic virus phenotype is determined by three or more V (virus-inducing) loci. A single locus for inducibility of xenotropic murine leukemia virus was mapped to chromosome 1 close to, but possibly not allelic to, Bxv-1. Although the high ecotropic virus phenotype is phenotypically dominant, the high xenotropic virus phenotype was recessive in all crosses tested. Suppression of xenotropic murine leukemia virus is governed by a single gene which is not linked to the xenotropic V locus. PMID- 6286990 TI - Site-specific phosphorylation regulates the transcriptive activity of vesicular stomatitis virus NS protein. AB - In vitro transcription by vesicular stomatitis virus nucleocapsids is inhibited by enzymatic dephosphorylation of the NS protein. We provide evidence that specific, partial dephosphorylation of NS molecules is the only detectable change in nucleocapsids treated with bacterial alkaline phosphatase under conditions that prevent the action of adventitious protease. Dephosphorylation appeared to affect only the rate of transcription; there were no changes in sedimentation rates of transcripts. To identify the sites of phosphorylation required for NS activity in transcription, we examined phosphopeptides produced by chymotrypsin digestion of the two electrophoretic classes of NS molecules found in virions and infected cells. The electrophoretically slower class, NS1, abundant in the intracellular soluble pool, has a lower activity in transcription; it contained six chymotryptic phosphopeptides. Five of these peptides contained both phosphoserine and phosphothreonine, indicating that this peptide cluster represents at least 11 separate sites of phosphorylation. In the electrophoretically faster nucleocapsid-associated NS2 class of molecules, which support a higher rate of transcription, another group of eight phosphopeptides was superimposed on this pattern. Two of these peptides contained both phosphoserine and phosphothreonine, so this cluster of peptides represents at least 10 additional phosphorylation sites. These sites were especially sensitive to dephosphorylation by bacterial alkaline phosphatase. One or more of them appears to be responsible for the higher transcription rates medicated by NS2 molecules. PMID- 6286991 TI - Molecular cloning and characterization of murine leukemia virus-related DNA sequences from C3H/HeN mouse DNA. AB - Ten murine leukemia virus (MuLV)-related DNA sequences were isolated from C3H/HeN mouse genomic DNA by cloning of EcoRI fragments in a Charon 4A vector. Detailed restriction endonuclease maps of four of the clones were developed by using AKR MuLV [32P]cDNA as a probe. C3H clone 14-9 contains approximately 7 kilobase pairs of MuLV-related DNA, one copy of an MuLV long terminal repeat-like sequence, and a region of flanking mouse DNA. C3H clones 34.2 and 36.1 contain approximately 2 kilobase pairs of MuLV-related DNA, one copy of a MuLV LTR-like sequence, and differing lengths of flanking mouse DNA sequences. C3H clone 8.13 was found to contain an insert of 5.7 kilobase pairs of MuLV-related DNA with two long terminal repeat-like regions and sequences which are partially homologous to AKv 1. Comparison fo the restriction endonuclease cleavage maps of these C3H clones with maps recently developed for ecotropic and xenotropic MuLV DNAs indicates that C3H clone 14-9 corresponds to the 5'-terminal portion of a genomic DNA sequence related to xenotropic MuLVs, whereas C3H clones 34.2 and 36.1 correspond to the 3' terminal portions of genomic DNA sequences related to xenotropic MuLVs. Clone 8.13 represents a deleted, xenotropic MuLV-related provirus. C3H clones 14 9, 34.2, 36.1, and 8.13 provide defined DNA sequence probes with which to characterize the organization and expression of endogenous MuLV-related DNA sequences in the mouse genome. PMID- 6286992 TI - Most of the murine leukemia virus sequences in the DNA of NIH/swiss mice consist of two closely related proviruses, each repeated several times. AB - The structure of the endogenous murine leukemia virus (MuLV) sequences of NIH/Swiss mice was analyzed by restriction endonuclease digestion, gel electrophoresis, and hybridization to an MuLV nucleic acid probe. Digestion of mouse DNA with certain restriction endonucleases revealed two classes of fragments. A large number of fragments (about 30) were present at a relatively low concentration, indicating that each derived from a sequence present once in the mouse genome. A smaller number of fragments (one to five) were present at a much higher concentration and must have resulted from sequences present multiple times in the mouse genome. These results indicated that the endogenous MuLV sequences represent a family of dispersed repetitive sequences. Hybridization of these same digested mouse DNAs to nucleic acid probes representing different portions of the MuLV genome allowed construction of a map of the sites where restriction endonucleases cleave the endogenous MuLV sequences. Several independent recombinant DNA clones of endogenous MuLV sequences have been isolated from C3H mice (Roblin et al., J. Virol. 43:113-126, 1982). Analysis of these sequences shows that they have the structure of MuLV proviruses. The sites at which restriction endonucleases cleave within these proviruses appeared to be similar or identical to the sites at which these nucleases cleaved within the MuLV sequences of NIH/Swiss mice. This identity was confirmed by parallel electrophoresis. We conclude that the apparently complex pattern of endogenous MuLV sequences of NIH/Swiss mice consists largely of only two kinds of provirus, each repeated multiple times at dispersed sites in the mouse genome. PMID- 6286993 TI - Restriction enzyme mapping of vaccinia virus DNA. AB - The cleavage sites for the restriction enzymes Bg/I, HindIII, KpnI, SalI, SmaI, and XhoI were located, from primary data, on the DNA isolated from the WR strain of vaccinia virus. Bg/I and SmaI divide the DNA into five segments which can be isolated by sucrose gradient centrifugation. These large segments provide a convenient means to group segments produced by other enzymes. The construction of physical maps was initiated by identifying the segments at each end of the DNA and then finding segments which were adjacent to these terminal sections. This was accomplished by isolating large shear fragments which contained the covalently linked termini of the DNA. Most of the data needed to derive the maps were obtained by isolating segments produce by one enzyme and then cleaving these individual segments with a second enzyme. PMID- 6286994 TI - Identification of a unique mouse mammary tumor virus in the BALB/cNIV mouse strain. AB - We examined the genetic structure, in terms of restriction endonuclease recognition sites, of the milk-transmitted, low-oncogenic mouse mammary tumor virus (MuMTV) of the BALB/cNIV mouse strain. An analysis with EcoRI documented the presence of acquired cNIV proviruses in the mammary tumor DNAs of BALB/cNIV animals. A comparison of tumor DNAs digested with PstI showed that both the cNIV MuMTV and C3Hf MuMTV proviruses lacked the 4.3- and 1.1-kilobase pair fragments characteristic of C3H MuMTV patterns. An examination of mammary tumor and normal, nonmammary tissue DNAs with BamHI supported the idea that the cNIV MuMTV is identical to the C3Hf MuMTV and demonstrated that these two low-oncogenic proviruses are identical to the high-oncogenic C3H MuMTV provirus with respect to a pair of BamHI sites which define a 1.3-kilobase pair fragment. For each of the three MuMTV strains, we also mapped DNAs generated in isolated virions by reverse transcription of their genomic RNAs. Our results showed that cNIV and C3Hf MuMTV are distinct entities by virtue of an additional PstI site within the cNIV long terminal repeat sequence. Another unique feature of cNIV MuMTV revealed by the analysis of virion-generated DNAs was the existence of a family of genomes within the cNIV population. We concluded that cNIV is distinct from its presumptive C3Hf MuMTV predecessor. PMID- 6286995 TI - In vitro transcription of vesicular stomatitis virus: initiation with GTP at a specific site within the N cistron. AB - In vitro transcripts of vesicular stomatitis virus (VSV) were either 5' terminally labeled by incorporation of [beta-(32)P]GTP or were selected on Hg agarose after incorporation of gamma-thio-GTP. Capped RNAs ranged in size from 23 nucleotides, the shortest capped RNA detected, to full-length message size. The 5'-terminal sequences corresponded to those of N message and to a small amount of NS message. Approximately 14% of the capped N gene transcripts were terminated at positions 86 to 90 of the VSV genome, giving rise to specific, 36 to 40 nucleotide-long, capped RNA species. The GTP-initiated RNAs were short with a predominant 28-nucleotide-long RNA species. A minor portion was as large as mRNAs. Nucleotide sequence analyses of the short RNA revealed that it was specifically initiated at positon 91 of the VSV genome, 41 nucleotides within the N cistron. This corresponds exactly to the site where transcription of the 40 nucleotide-long, capped RNA terminated. Initiation with GTP at position 91 occurred at approximately the same frequency as termination of the capped RNA at position 90, suggesting that intracistronic initiation at position 91 may depend upon termination of transcription of the 5'-proximal region and therefore may be sequential. This unique RNA represents the first transcript of VSV which was initiated at an intracistronic site with GTP, and may also represent the first example of a transcript derived from a stop/start mechanism of VSV transcription in vitro. Although initiation occurred frequently at the beginning of the N cistron yielding 11 to 14-nucleotide-long, [beta-(32)P]ATP-labeled transcripts (D. F. Pinney and S. U. Emerson, J. Virol. 42:889-896, 1982), capping of these short RNAs was not detected. This suggests that transcripts may have to be 15 to 23 nucleotides long to be accepted as substrates by the guanyltransferase. PMID- 6286996 TI - Isolation and characterization of defective simian virus 40 genomes which complement for infectivity. AB - A new variant of simian virus 40 (EL SV40), containing the complete viral DNA separated into two molecules, was isolated. One DNA species contains nearly all of the early (E) SV40 sequences, and the other DNA contains nearly all of the late (L) viral sequences. Each genome was encircled by reiterated viral origins and termini and migrated in agarose gels as covalently closed supercoiled circles. EL SV40 or its progenitor appears to have been generated in human A172 glioblastoma cells, as defective interfering genomes during acute lytic infections, but was selected during the establishment of persistently infected (PI) green monkey cells (TC-7). PI TC-7/SV40 cells contained EL SV40 as the predominant SV40 species. EL SV40 propagated efficiently and rapidly in BSC-1, another line of green monkey cells, where it also formed plaques. EL SV40 stocks generated in BSC-1 cells were shown to be free of wild-type SV40 by a number of criteria. E and L SV40 genomes were also cloned in the bacterial plasmid pBR322. When transfected into BSC-1 cell monolayers, only the combination of E and L genomes produced a lytic infection, followed by the synthesis of EL SV40. However, transfection with E SV40 DNA alone did produce T-antigen, although at reduced frequency. PMID- 6286997 TI - Sequence of the long terminal repeat and adjacent segments of the endogenous avian virus Rous-associated virus 0. AB - Rous-associated virus 0 (RAV-0), an endogenous chicken virus, does not cause disease when inoculated into susceptible domestic chickens. An infectious unintegrated circular RAV-0 DNA was molecularly cloned, and the sequence of the long terminal repeat (LTR) and adjacent segments was determined. The sequence of the LTR was found to be very similar to that of replication-defective endogenous virus EV-1. Like the EV-1 LTR, the RAV-0 LTR is smaller (278 base pairs instead of 330) than the LTRs of the oncogenic members of the avian sarcoma virus-avian leukosis virus group. There is, however, significant homology. The most striking differences are in the U(3) region of the LTR, and in this region there are a series of small segments present in the oncogenic viruses which are absent in RAV 0. These differences in the U(3) region of the LTR could account for the differences in the oncogenic potential of RAV-0 and the avian leukosis viruses. I also compared the regions adjacent to the RAV-0 LTR with the available avian sarcoma virus sequences. A segment of approximately 200 bases to the right of the LTR (toward gag) is almost identical in RAV-0 and the Prague C strain of Rous sarcoma virus. The segment of RAV-0 which lies between the end of the env gene and U(3) is approximately 190 bases in length. Essentially this entire segment is present between env and src in the Schmidt-Ruppin A strain of Rous sarcoma virus. Most of this segment is also present between env and src in Prague C; however, in Prague C there is an apparent deletion of 40 bases in the region adjacent to env. In Schmidt-Ruppin A, but not in Prague C, about half of this segment is also present between src and the LTR. This arrangement has implications for the mechanism by which src was acquired. The region which encoded the gp37 portion of env appears to be very similar in RAV-0 and the Rous sarcoma viruses. However, differences at the very end of env imply that the carboxy termini of RAV-0, Schmidt-Ruppin A, and Prague C gp37s are significantly different. The implications of these observations are considered. PMID- 6286998 TI - Organization of the Epstein-Barr virus DNA molecule. II. Fine mapping of the boundaries of the internal repeat cluster of B95-8 and identification of additional small tandem repeats adjacent to the HR-1 deletion. AB - We used cloned BamHI fragments from Epstein-Barr virus strain B95-8 [EBV(B95 8)]DNA to obtain detailed restriction maps of the region of the genome adjacent to the large internal repeat cluster. These maps together with the results of hybridization experiments using a 3.1-kilobase repeat probe defined more precisely the location of the injection between the internal repeat cluster and the flanking unique-sequence DNA. On one side (UL), the repeat sequences extended 600 +/- 80 base pairs (bp) into BamHI-Y; on the other side (US), they extended 1,300 +/- 200 bp into BamHI-C. Therefore, EBV(B95-8) DNA contained a nonintegral number of 3.1-kilobase repeat units, namely, 12.6 copies. The mapping studies also revealed a second series of internal tandem repetitions in EBV(B95-8) DNA located within the BamHI-H fragment. This cluster comprised 11 copies of a 135-bp repeat unit which contained a single site for the NotI restriction endonuclease. Hybridization to these cloned EBV(B95-8) fragments using total EBV(HR-1) DNA as probe indicated that the deletion in EBV(HR-1) removed all 3,000 bp of unique sequence DNA which lay between the large 3.1-kilobase and the small 135-bp repeat clusters. Thus, the deletion which destroyed the transforming ability in the EBV(HR-1) virus was bounded on either side by tandem repetitions. PMID- 6286999 TI - RNA polymerase-associated interactions near template promoter sequences of defective interfering particles of vesicular stomatitis virus. AB - Methylation protection studies suggested that the NS protein component of the RNA polymerase of vesicular stomatitis virus contacts the RNA templates of defective interfering (DI) particles at the sequence 3'...GUCUAUUUUUAUUUUUGGUG...5',17 to 37 nucleotides downstream from the site of initiation of in vitro transcription. The data indicated that vesicular stomatitis virus and DI particle RNAs contain different polymerase binding sequences and that NS may function as a transcription initiator protein for template recognition at both sequences. These results are thus compatible with the hypothesis that differences in the rate of defective and nondefective viral particle replication and autointerference are due to higher-affinity binding sites for polymerase at the 3' end of DI particle RNAs. In addition, a unique DI particle (DI-LT2) RNA that contains a transcriptionally inactive vesicular stomatitis virus leader gene 72 to 118 nucleotides from its 3' end showed interactions with the viral polymerase similar to those reported previously for the 3'-terminal vesicular stomatitis virus leader gene (Keene et al., Proc. Natl. Acad. Sci, U.S.A. 78:6191--6195, 1981). The interaction of polymerase with the internal leader gene of DI-LT2 RNA suggested that the lack of leader RNA and mRNA production by this particle is not due to the inability of polymerase to bind to internal sites along the template. Instead, the initiation of transcription is more likely influenced by the position of the polymerase binding site relative to the 3' end or by requisite interactions between the catalytic polymerase component (L) and the proposed initiator protein (NS). PMID- 6287000 TI - Inhibition of host translation in encephalomyocarditis virus-infected L cells: a novel mechanism. AB - Encephalomyocarditis virus induced a rapid shutoff of host translation in mouse L cells shortly after infection and before viral proteins were made in detectable amounts. This kinetic pattern is similar to that seen in poliovirus-infected HeLa cells. However, the mechanisms of host shutoff are different in these two cases, for no reduction in the ability of lysates from encephalomyocarditis virus infected L cells to translate capped mRNAs was observed. Instead, a change in the subcellular distribution of one or more initiation factors was seen. In particular, cap recognition activity in the high-speed supernatant fraction (S200) prepared from cell lysates increased threefold as a result of virus infection. The significance of this observation in terms of possible shutoff mechanisms is discussed. Inasmuch as the rapid host shutoff is not induced in at least four other cell types by encephalomyocarditis virus infection, it may be concluded that host shutoff mechanisms not only vary within the picornavirus group, but also depend upon the particular cell type employed. PMID- 6287001 TI - Organization, distribution, and stability of endogenous ecotropic murine leukemia virus DNA sequences in chromosomes of Mus musculus. AB - The endogenous ecotropic murine leukemia virus DNA content and integration sites were characterized for 54 inbred strains and substrains of mice by restriction enzyme digestion, Southern blotting, and hybridization with an ecotropic murine leukemia virus DNA-specific probe. More than 75% of these strains carried endogenous ecotropic proviruses which were located in at least 29 distinct integration sites in chromosomes of Mus musculus. Fourteen of these proviruses have been assigned specific locus designations. Most, but not all, of the endogenous ecotropic proviruses were structurally indistinguishable by this analysis from the prototype AKR ecotropic virus, and the distribution of these proviruses followed known relationships among the inbred strains and substrains of mice. These results suggest that, in general, viral DNA integration preceded the establishment of inbred mouse strains and that these integrations are relatively stable. PMID- 6287002 TI - Synthesis of virus-specific polypeptides and genomic RNA during the replicative cycle of Pichinde virus. AB - A stock of plaque-purified Pichinde virus, prepared under conditions designed to limit the amounts of defective interfering virus, was used to infect BHK cells. At daily intervals after infection, cells were examined for infectious and radiolabeled virus particle production and for the synthesis of virus-specific polypeptides. Quantitative comparisons were also made of the concentrations of genomic Pichinde virus L and S RNAs in the cytoplasm of infected cells on different days after infection. Our results showed that virus particle production, rates of protein synthesis, and the intracellular levels of viral genomic RNAs all increased and decreased with similar kinetics, and that this regulation was independent of the cell growth cycle. We were unable to relate these changes in viral macromolecule and virus production to the appearance of readily identifiable defective interfering particles. Our findings suggest that regulation of virus replication early during the replicative cycle of Pichinde virus may not be dependent upon the generation of defective interfering virus. PMID- 6287003 TI - Monoclonal antibodies to the p21 products of the transforming gene of Harvey murine sarcoma virus and of the cellular ras gene family. AB - We have isolated eight rat lymphocyte-myeloma hybrid cell lines producing monoclonal antibodies that react with the 21,000-dalton transforming protein (p21) encoded by the v-ras gene of Harvey murine sarcoma virus (Ha-MuSV). These antibodies specifically immunoprecipitate both phosphorylated and non phosphorylated forms of p21 from lysates of cells transformed by Ha-MuSV. All eight react with the products of closely related ras genes expressed in cells transformed by two additional sarcoma viruses (rat sarcoma virus and BALB sarcoma virus) or by a cellular Harvey-ras gene placed under the control of a viral promoter. Three of the antibodies also react strongly with the p21 encoded by the v-ras gene of Kirsten MuSV. These same three antibodies immunoprecipitate the predominant p21 species synthesized normally in a variety of rodent cell lines, including the p21 produced at high levels in 416B murine hemopoietic cells. This suggests that an endogenous gene closely related to Kirsten-ras is expressed in these cells. The monoclonal antibodies have been used to confirm two properties associated with p21; localization at the inner surface of the membrane of Ha-MuSV transformed cells, assayed by immunofluorescence microscopy, and binding of guanine nucleotides. PMID- 6287004 TI - Filamentous structures associated with Epstein-Barr virus-infected cells. AB - After the onset of Epstein-Barr virus DNA and protein synthesis 10 h after superinfection of Raji cells (a cell line containing Epstein-Barr virus DNA but not producing virus), filamentous structures 25 nm in diameter and 0.2 to 1.4 micrometers in length could be detected in the cell cytoplasm by electron microscopy. These structures banded in metrizamide gradients with viral DNA and proteins, but at a density different from that of virions or nucleocapsids. These filaments, enriched in a 155,000-dalton protein similar in size to a major nucleocapsid protein of Epstein-Barr virus, may represent intermediates in viral nucleocapsid assembly. PMID- 6287005 TI - Definition of a series of stages in the association of two herpesviral proteins with the cell nucleus. AB - We examined the kinetics and the nature of the association of two herpes simplex virus proteins, the major DNA-binding protein (ICP8) and the major capsid protein (ICP5), with the nuclei of infected cells. We defined a series of stages in the association of the ICP8 protein with the cell nucleus. (i) Immediately after synthesis, the protein was found in the cytoplasmic fraction but associated rapidly with the crude nuclear fraction. (ii) The initial association of ICP8 with the crude nuclear fraction was detergent sensitive but DNase resistant, and, thus, the protein was either bound to structures attached to the outside of the nucleus and had not penetrated the nuclear envelope or was loosely bound in the nucleus, (iii) At intermediate times, a low level of an intermediate form was observed in which the association of ICP8 with the nuclear fraction was resistant to both detergent and DNase treatment. The protein may be bound to the nuclear matrix at this stage. Inhibition of viral DNA synthesis caused the DNA-binding protein to accumulate in this form. (iv) At late times during the chase period, the association of ICP8 with the cell nucleus was resistant to detergent treatment but sensitive to DNase treatment. our results argue that at this stage ICP8 was bound to viral DNA. Thus, nuclear association of the DNA-binding protein did not require viral DNA replication. More important is the observation that there is a series of stages in the nuclear association of this protein, and, thus, there may be a succession of binding sites for this protein in the cell during its movement to its final site of action in the nucleus. The major capsid protein showed some similar stages of association with the cell nucleus but the initial association with the nucleus followed a lag period. Its early association with the crude nuclear fraction was also detergent sensitive but was resistant to detergent treatment at later times. Its association with the cell nucleus was almost completely resistant to DNase treatment at all times. Inhibition of viral DNA replication blocked the nuclear transport of this protein. Thus, these two viral proteins share some stages in nuclear transport, although their requirements for nuclear association are different. PMID- 6287006 TI - Differential effect of nucleoside analog triphosphates on ribonucleotide reductases from uninfected and herpes simplex virus-infected HeLa cells. AB - Effects of the triphosphates of eight pyrimidine nucleoside analogs (5 substituted, 2'-fluoroara-, and acyclonucleosides) and acycloguanosine were examined on the ribonucleotide reductases prepared from uninfected and herpes simplex virus (types 1 and 2)-infected HeLa cells. Of the analogs tested, E-5 propenyl- and E-5-(2-bromovinyl)-dUTP were more potent inhibitors than dTTP of the enzymes from virus-infected cells, whereas only the former compound showed this effect on the uninfected HeLa enzyme. PMID- 6287007 TI - Isolation and characterization of simian virus 40 early region deletion mutants. AB - We constructed a tsB4/dl884 double-mutant helper virus and used it to isolate two simian virus 40 early region deletion mutants that lack about half of the DNA sequences normally used to encode the large tumor antigen (T). Both mutants make a normal-sized small t antigen, but neither mutant can replicate its DNA in the absence of a T+ helper. PMID- 6287009 TI - Improved infectivity of reassembled polyoma virus. AB - Polyoma virus was dissociated into capsomeres (18, 12, and 5S) and a DNA-protein complex (48S) with the Ca2+ chelator, ethyleneglycol-bis-N,N'-tetraacetic acid, and the reducing agent, 2-mercaptoethanol. The reaction was maintained at pH 5.0. Reassembly of the dissociated components to complete virions was accomplished by dialyzing these components overnight at 4 degrees C against the reassembly buffer containing CaCl2, dimethylsulfoxide, Triton X-100, and 0.01 M Tris-acetic acid (pH 5.0). Reconstructed particles ranged from 240S complete virions to lighter intermediate species. Approximately 25% of the dissociated particles could be physically reassembled to complete virions. These virions regained 12.5% of their hemagglutination ability and as much as 6.7% of their original infectivity. The infectivity of these reassembled particles represented a 100-fold increase in infectivity compared with that of the particles that were dissociated and reassembled at pH 7.4. Biochemical analysis showed that the polyoma viral receptor of the virions reassembled at pH 7.4 was greatly reduced, whereas virions reassembled at pH 5.0 retained their receptor. Reassembly could be further improved by additions of either exogenous capsomeres or DNA-protein complex to the reassembly reaction mixture. PMID- 6287008 TI - Inhibitory effect of E-5-(2-bromovinyl)-1-beta-D-arabinofuranosyluracil on herpes simplex virus replication and DNA synthesis. AB - The effect of E-5-(2-bromovinyl)-1-beta-D-arabinofuranosyluracil (BVaraU) on herpes simplex virus (HSV) replication was examined and compared with that of E-5 (2-bromovinyl)-2'-deoxyuridine (BVdUrd). The 50% inhibitory dose against HSV type 1 (HSV-1) was 0.1 microgram/ml compared with 0.008 microgram/ml for BVdUrd; the antimetabolic 50% inhibitory dose of BVaraU ranged from 20 to 95 micrograms/ml. The addition of 50 micrograms of BVaraU per ml to HSV-1-infected Vero cells decreased the synthesis of viral and cellular DNA by 37 and 28%, respectively. The 5'-triphosphate (BVaraUTP) competed with dTTP in DNA synthesis by the herpes viral and cellular DNA polymerases; the apparent Ki values of HSV-1 DNA polymerase, DNA polymerase alpha, and DNA polymerase beta were 0.14, 0.32, and 5 microM, respectively. Thus, BVaraU was a less effective antiherpesvirus agent than BVdUrd; unlike BVdUrd, it did not appear to be internally incorporated into replicating DNA in virus-infected cells. PMID- 6287010 TI - Identification of a DNA fragment from a molecularly cloned mink cell focus inducing murine leukemia virus specific for xenotropic virus-related sequences. AB - Molecular clones of closed circular DNA molecules of a mink cell focus-inducing murine leukemia virus (MCF-13 MuLV) were generated. Closed circular DNA molecules isolated from a Hirt extraction of recently infected NIH/3T3 cells were inserted at their unique EcoRI site into lambda gtWES.lambda B. Restriction endonuclease analysis of inserts of two clones indicated that they represented intact MCF-13 MuLV genomes. One viral insert contained two large terminal repeat sequences, and the other contained only one. A 300-base-pair DNA fragment located in the envelope region of the MCF-13 MuLV genome was determined to be related to xenotropic MuLV sequences. PMID- 6287011 TI - Herpesvirus saimiri strain variability. AB - Herpesvirus saimiri was isolated from 22 squirrel monkeys by cocultivation of peripheral lymphocytes with permissive owl monkey kidney monolayer cells. Comparison of virion DNA fragments produced from restriction endonuclease digestion was used as a sensitive measure of strain variability. Although all isolates contained similarities and common features, 19 of the 22 were readily distinguished. Three of the isolates, however, were indistinguishable and possibly were related epidemiologically. Distinct subtypes of H. saimiri were not evident by these criteria; Peruvian, Colombian, Guyanan, and Bolivian squirrel monkeys yielded isolates without characteristic features peculiar to the geographic region. Three of three colony-born squirrel monkeys that were tested yielded a strain of virus distinct from that obtained from the mother. In separate experiments, two of three animals chosen at random yielded a strain of virus different from that originally obtained 16 and 22 months previously; only one of the three animals examined yielded the same strain of virus 22 months after the original isolation. The degree of restriction endonuclease fragment variability among H. saimiri strains appeared to be greater than previously observed for other herpesviruses. PMID- 6287013 TI - Homology between the glycoproteins of vesicular stomatitis virus and rabies virus. AB - We compared the predicted amino acid sequences of the vesicular stomatitis virus and rabies virus glycoproteins by using a computer program which provides an optimal alignment and a statistical significance for the match. Highly significant homology between these two proteins was detected, including identical positioning of one glycosylation site. A significant homology between the predicted amino acid sequences of vesicular stomatitis virus and influenza virus matrix proteins was also found. PMID- 6287012 TI - Polysome-associated proteins in herpes simplex virus-infected cells. AB - In the cytoplasm of eucaryotic cells, mRNA is associated with proteins. These mRNA-protein complexes, termed messenger ribonucleoprotein (mRNP) particles, are divided into two functional classes. The first class contains free (non-ribosome associated) mRNPs which have been termed informosomes by others. The second class of mRNPs, those associated with polysomes, are actively engaged in protein synthesis and are termed polysomal mRNPs. The experiments described in this paper examined the proteins associated with polyribosomes in uninfected and herpes simplex virus type 1-infected cells. The data indicate that after infection with herpes simplex virus type 1, specific changes occur in the proteins which normally are found associated with these polysomal mRNPs. These changes include both the appearance of new and possibly virus-specific proteins and the loss of normal host-specific proteins. The relationship of these changes to the patterns of protein synthesis in these cells is also discussed. PMID- 6287014 TI - Use of a hybrid infectivity assay to analyze primary transcription of temperature sensitive mutants of the New Jersey serotype of vesicular stomatitis virus. AB - A hybrid infectivity assay specific for primary transcription was developed to analyze the production of functional mRNAs by vesicular stomatitis virus. A template prepared from wild-type virions of the New Jersey serotype of vesicular stomatitis virus was reconstituted with RNA polymerase proteins from the wild type or temperature-sensitive mutants, and the in vivo temperature sensitivity of the polymerase was determined by infectivity assay. The data demonstrate that the New Jersey temperature-sensitive mutants A1 and E1 have non-temperature-sensitive transcriptases, whereas the B1 and F1 mutants both have temperature-sensitive L proteins which are defective in primary transcription. PMID- 6287016 TI - Patterns of genomic distribution and sequence heterogeneity of a murine "retrovirus-like" multigene family. AB - The mouse genome contains over 100 copies of a dispersed gene family known as "virus-like" genes encoding 30S RNA (VL30). Although they do not share nucleotide sequence homology with known retroviruses, these genetic elements are distinguished by several "retrovirus-like" features, notably, the capacity of the 30S RNA transcripts of these genes to be encapsidated by c-type virions and the transmissibility of VL30 information to other cells via pseudovirion infection. Using VL30 DNA units, cloned from the BALB/c mouse embryonic gene library, we have recently shown that VL30 DNA units share basic structural features with retrovirus proviruses. To shed light on the relatedness of VL30 information to endogenous proviruses and possibly other genetic elements, we extended our previous studies concerning genomic distribution patterns of VL30 elements and patterns of sequence heterogeneity among VL30 units. The following observations were made: (i) VL30 units were distributed among different mouse chromosomes; (ii) distribution patterns of VL30 units markedly differed among mouse strains; (iii) there was constancy of VL30 restriction patterns in different tissues; (iv) a high degree of sequence divergence existed among different VL30 units cloned from the same embryo; and (v) VL30 units were heterogeneous with respect to the state of DNA methylation. The results are discussed in terms of the possible modes of evolution of this multigene family. PMID- 6287015 TI - Location of the structural genes for glycoproteins gD and gE and for other polypeptides in the S component of herpes simplex virus type 1 DNA. AB - To map the structural genes for the gD and gE polypeptides and for other viral products encoded in the S component of herpes simplex virus type 1 DNA, we selected mRNAs capable of hybridizing to cloned viral DNA fragments and translated the mRNAs in vitro to determine which polypeptides were encoded therein. The gD and gE polypeptides were identified by immunoprecipitation with appropriate monoclonal and monospecific antibodies, whereas the other polypeptides were characterized only by their electrophoretic mobilities in polyacrylamide gels. We found that gD mRNA hybridized to a single SacI subfragment of BamHI fragment J, whereas gE mRNA hybridized to an adjacent SacI subfragment of BamHI fragment J and also to BamHI fragment X. These and other results permit the conclusion that the structural gene for gD is located between map coordinates 0.911 and 0.924, and the gene for gE is between map coordinates 0.924 and 0.951. We also found that mRNAs for polypeptides of 55,000, 42,000, 33,000, and 22,000 molecular weight hybridized to DNA fragments spanning the regions from map coordinates 0.911 to 0.924, 0.897 to 0.911, 0.939 to 0.965, and 0.939 to 0.965, respectively. Finally, in accord with the results of others, we found that mRNA for a 68,000-molecular-weight polypeptide hybridized to the two noncontiguous BamHI fragments N and Z, which share a reiterated DNA sequence. PMID- 6287017 TI - Discrete regions of simian virus 40 large T antigen are required for nonspecific and viral origin-specific DNA binding. AB - The nondefective adenovirus type 2 (Ad2)-simian virus 40 (SV40) hybrid viruses, Ad2+ND2 and Ad2+ND4, have been used to determine which regions of the SV40 genome coding for the large tumor (T) antigen are involved in specific and nonspecific DNA binding. Ad2+ND2 encodes 45,000 M4 (45K) and 56,000 Mr (56K) T antigen related polypeptides. The 45K polypeptide did not bind to DNA, but the 56K polypeptide bound nonspecifically to calf thymus DNA, Ad2+ND4 encodes 50,000 Mr (60K), 66,000 Mr (66K), 70,000 Mr (70K), 74,000 Mr (74K), and 90,000 Mr (90K) T antigen-related polypeptides, all of which bound nonspecifically to calf thymus DNA. However, in more stringent assays, where tight binding to viral origin sequences was tested, only the 90K protein specified by Ad2A+ND4 showed specific high affinity for sequences at the viral origin of replication. From these results and previously published experiments describing the SV40 DNA integrated into these hybrid viruses, it was concluded that SV40 early gene sequences located between 0.39 and 0.44 SV40 map units contribute to nonspecific DNA binding, whereas sequences located between 0.50 and 0.63 SV40 map units are necessary for specific binding to the viral origin of replication. PMID- 6287018 TI - Internal organization of endogenous proviral DNAs of xenotropic murine leukemia viruses. AB - The internal organization of endogenous xenotropic murine leukemia virus proviruses was determined in a series of blot hybridization experiments in which DNA from several different inbred mouse strains, digested with restriction enzymes known to cleave xenotropic proviral DNAs at least twice, was annealed to generalized murine leukemia virus or xenotropic env-specific DNA probes. Comigrating bands of variable intensity which hybridized to the xenotropic env probe were identified in all inbred mouse DNA preparations. At least seven classes of endogenous xenotropic proviral DNA with respect to SacI cleavage maps were detected in mouse DNA. Two of the seven classes were indistinguishable from proviruses associated with known infectious xenotropic murine leukemia viruses. These results are consistent with the existence of related but organizationally distinct families of endogenous xenotropic proviral DNA that are present in different relative abundances in mouse genomic DNA. PMID- 6287021 TI - Sequence diversity of human rotavirus strains investigated by northern blot hybridization analysis. AB - Rotavirus genomic RNAs, derived from a series of human isolates that exhibit variability in the pattern of migration of the double-stranded RNA on polyacrylamide gels, were transferred to diazobenzyloxymethyl paper, and their sequence diversity was investigated. Hybridization of cDNA probes prepared from the 11 segments of rotavirus RNA indicated that considerable sequence diversity exists among these viruses. Under conditions of both low and high stringency, hybridization analysis of virus collected between 1975 and 1980 suggested that the variation among rotavirus strains may have occurred by a process involving both "drift" and "shift" in the sequence of the rotavirus genomic segments. PMID- 6287019 TI - Transformation of NIH 3T3 cells with cloned fragments of human cytomegalovirus strain AD169. AB - NIH 3T3 cells were transfected with restriction endonuclease and cloned human cytomegalovirus DNA fragments to identify the transforming region(s). Cleavage of human cytomegalovirus strain AD169 DNA with XbaI and HindIII left a transforming region intact whereas EcoRI inactivated this function. Transfection of cells with cosmids containing human cytomegalovirus DNA spanning the entire genome resulted in transformation by one cosmid, pCM1058, with the AD169 HindIII DNA fragments E, R, T, and a'. Cells were selected for their growth in 1.2% methylcellulose. The clones isolated had a significant replating efficiency and were oncogenic in BALB/c nu/nu mice. Transfection of cosmids and plasmids containing subsets of the viral sequences in pCM1058 identified a common region possessed by all of the transforming recombinant molecules. This region was in the HindIII E fragment with the left boundary defined by the EcoRI d-R junction and the right boundary defined by the HindIII E-T junction. Further mapping and transfection experiments determined that the transforming region was contained without a 2.9-kilobase fragment between map units 0.123 and 0.14 on the prototype molecule of the AD169 strain. PMID- 6287020 TI - Loss of viral gene expression and retention of tumorigenicity by Abelson lymphoma cells. AB - Lymphomas induced by the Abelson murine leukemia virus (A-MuLV) were examined for the expression of biochemical and biological markers associated with A-MuLV transformation before and after in vivo growth in genetically distinguishable host mice. Although all tumors and clonal lines derived from them initially expressed the A-MuLV-encoded gag fusion protein p160, they ceased synthesis of this molecule after several weeks of growth in vivo as ascites tumors. Transplanted clonal lines continued to express the alloantigenic marker H-2b and the isoenzyme marker Gpi-1b of the donor tumor cells, indicating that the cells were of donor and not host origin. Examination of cellular DNA obtained from p160 positive and derivative p160-negative lines indicated that p160-negative clones had lost A-MuLV-specific proviral sequences as detected by hybridization with several probes. Although the clonal lines no longer expressed p160, they retained their malignant phenotype and continued to express the Abelson antigen, a cell surface marker associated with A-MuLV lymphomagenesis. Continued expression of the A-MuLV genome was not required for maintenance of oncogenic potential under these conditions of in vivo tumor growth. PMID- 6287022 TI - Sequence relationships between the genome segments of human and animal rotavirus strains. AB - The sequence relationships of a range of cultivable and noncultivable human and animal rotaviruses were investigated by hybridization of rotavirus cDNA probes to genomic RNAs immobilized on diazobenzyloxymethyl paper. Under conditions of low stringency (34% base mismatch tolerated) most genome segments exhibited partial homology except for genes 4 and 5. In contrast, under more stringent conditions of hybridization in which no more than 8% base mismatch was tolerated, few segments exhibited homology. Generally the human and animal rotaviruses were found to possess distinct nucleic acid sequences that exhibit only a low order of sequence relatedness. These results are consistent with the notion that both cumulative changes in nucleic acid sequences and the interchange of segments may be involved in the evolution of distinct rotavirus strains. PMID- 6287023 TI - mRNA- and DNA-directed synthesis of herpes simplex virus-coded exonuclease in Xenopus laevis oocytes. AB - Microinjection of herpes simplex virus (HSV)-infected cell mRNA into Xenopus laevis oocytes resulted in the production of a new exonuclease activity. This enzyme strongly resembled the HSV alkaline exonuclease in many biochemical properties, and hybrid-arrested translation studies showed that it was virus coded, mapping at 0.080 to 0.185 genome map units. Exonuclease mRNA had a size and genome location equivalent to the mRNA encoding V185 in reticulocyte lysates, suggesting that V185 is the exonuclease. The enzyme synthesized in oocytes was found to act as an exonuclease in vivo. Two plasmids containing HSV DNA fragments directed the synthesis of exonuclease when microinjected into oocyte nuclei, and this finding enabled the coding and control sequences for this gene to be localized to 0.155 to 0.185 genome map units. PMID- 6287024 TI - Mapping and identification of the vaccinia virus thymidine kinase gene. AB - The thymidine kinase gene of vaccinia virus (VV) was mapped on the viral genome by using cloned fragments of the viral DNA to hybridize to early viral mRNA. Individual DNA fragments that represented about half of the viral genome were assayed, both for their ability to arrest the cell-free synthesis of active VV thymidine kinase and for their ability to select functional mRNA for the viral enzyme. Both activities were located in HindIII fragment J, which maps near the middle of VV DNA and contains about 2.6% of the genome (4,800 base pairs). This DNA fragment encodes four known early polypeptides, and to determine which of these was thymidine kinase, early VV mRNA was fractionated by sucrose gradient centrifugation and used to direct cell-free synthesis of the active enzyme. The thymidine kinase mRNA cosedimented with several species that encoded polypeptides in the molecular weight range 15,000 to 25,000. Hybridization of these mRNAs to HindIII-J DNA selected a message that directed the synthesis of thymidine kinase and a single polypeptide with an apparent molecular weight of 19,000. The native molecular weight of VV thymidine kinase is about 80,000, so these data indicate that, unlike thymidine kinase from several other sources, the active VV enzyme is probably a tetramer of 19,000-molecular-weight subunits. PMID- 6287025 TI - Genetic alterations of RNA leukemia viruses associated with the development of spontaneous thymic leukemia in AKR/J mice. AB - T1-oligonucleotide fingerprinting and mapping were used to study the expression of RNA leukemia viruses in leukemic and preleukemic AKR/J mice, with techniques designed to minimize the loss or inadvertent selection of viruses in vitro before biochemical analysis. In leukemic animals, complex mixtures of ecotropic and mink tropic viruses were expressed. Unique but similar polytropic virus-like genomes were present in each tumor isolate. In preleukemic mice, viral isolates from the thymus that were grown on NIH3T3 fibroblasts contained genomes with non-Akv polytropic virus-related oligonucleotides. This phenomenon was not evident in fingerprints of viruses from the spleen and bone marrow of the same animals. Remarkably, the non-Akv oligonucleotides located in the 3' portion of the P15E gene, the U3 noncoding region, and the 5' part of the gp70 gene were often expressed independently. Our results suggest the following. (i) Recombinant viruses can be detected in the thymuses of young preleukemic AKR mice and increase in relative abundance with age. (ii) During in vivo generation of the recombinant leukemogenic viruses, the selection of polytropic virus-related sequences in the 3' part of p15E and the U3 region and the 5' portion of gp70 occurs independently. (iii) Independent biological properties encoded in the gp70 and p15E regions of env of the recombinant viruses may mediate viral selection or leukemogenicity. (iv) The leukemogenic polytropic viruses of AKR/J mice arise via genetic recombination involving at least three endogenous viral sequences. PMID- 6287026 TI - Biochemical characterization of two types of human papillomaviruses associated with epidermodysplasia verruciformis. AB - The DNAs of the human papillomaviruses (HPVs) associated with the benign lesions of two patients suffering from epidermodysplasia verruciformis (patients JD and JK) were analyzed by using 12 restriction endonucleases. None of the restriction endonucleases were one-cut enzymes for the HPV DNA obtained from patient JD, referred to as the prototypical HPV-5, whereas five of them were one-cut enzymes for the DNA of the major virus found in patient JK, referred to as HPV-9. The molecular cloning of the two fragments resulting from the cleavage of HPV-5 DNA by endonuclease HindIII and of the single fragment obtained after treatment of HPV-9 DNA with endonuclease BamHI was performed in Escherichia coli after the fragments were inserted in plasmid pBR322. A cleavage map of the two cloned genomes was constructed. Little sequence homology (4 to 5%) was detected between HPV-5 and HPV-9 DNAs by DNA-DNA hybridization experiments in liquid phase at saturation; this homology was reproducibly higher than that (2 to 3%) detected under the same conditions between these DNAs and HPV-1a DNA. In addition, blot hybridization experiments performed under stringent conditions showed no or little sequence homology between the DNAs of HPV-5 and HPV-9 and those of HPV prototypes of types 1, 2, 3, 4, and 7 associated with skin warts. These results confirm that HPV-5 and HPV-9 are two distinct HPV types. PMID- 6287027 TI - Charge microheterogeneity of the major capsid protein of polyoma virus. AB - The behavior in isoelectric focusing of the major capsid polypeptide VPI of several strains of polyoma virus was studied. Two previously recognized phenomena were reexamined, namely, (i) the separation of the VP1 polypeptide into multiple subspecies differing only slightly from each other in apparent isoelectric point and (ii) strain differences in the overall apparent net charge of the family of VP1 subspecies. It was found that the pattern of subspecies was reproducible when focusing was initiated from either the basic or acidic region of the gel, keeping the ampholyte mixture constant. However, individual subspecies were unstable, and labeled polypeptide could be shifted dramatically by either refocusing of separated subspecies or by altering the concentration of ampholytes. These findings suggest that protein-protein and protein-ampholyte interactions play an important role in the generation of this charge heterogeneity. The basis for the overall charge difference between the VP1 of 3049 virus and several other strains (lpD, lpS, ts59, and A2) was studied, using recombinant viruses constructed of specific sequences derived from 3049 and lpD genomes. The portion of the VP1 polypeptide carrying the altered charge could be mapped to the body of the molecule 3' to the HindIII site at 45.0 map units (3,918 base pairs). This clearly segregates the VP1 charge phenotype from the cyc phenotype of 3049 in which capsid proteins are overproduced and accumulate in the cytoplasm of infected cells. PMID- 6287028 TI - Polymorphism of the migration of double-stranded RNA genome segments of avian reoviruses. AB - A number of field isolates of avian reovirus were characterized by analysis of the migration pattern of their genomic double-stranded RNA (dsRNA) segments upon polyacrylamide gel electrophoresis. Comparison of the various isolates has demonstrated (i) no relationship between serotype and migration of any individual dsRNA segment, (ii) marked polymorphism of migration patterns of all dsRNA segments among isolates of the same serotype as well as among different serotypes, (iii) no correlation between genotype and disease state, (iv) less marked variability in migration pattern from isolates within a restricted geographic locale compared to isolates from distant locales, (v) the presence of a single genotype in local outbreaks of disease, and (vi) the relative invariant migration of several dsRNA segments among the avian reoviruses, one of which (S1) may serve to distinguish the avian from the mammalian reoviruses. PMID- 6287029 TI - Construction and characterization of viable deletion mutants of simian virus 40 lacking sequences near the 3' end of the early region. AB - Five viable deletion mutants of simian virus 40 (SV40) were prepared and characterized. These mutants lack 15 to 60 base pairs between map positions 0.198 and 0.218, near the 3' end of the early region of SV40 and extend further into the body of the A gene, encoding the large T antigen, than previously described deletion mutants. These mutants were isolated after transfection of monkey kidney CV-1p cells with full-sized linear DNA prepared by partial digestion of form I SV40 DNA with restriction endonucleases HinfI or MboII, followed by removal of approximately 25 base pairs of DNA from the 5' termini using lambda-5' exonuclease and purification of the DNA in agarose gels. Based on camparisons of the DNA sequence of SV40 and polyoma virus, these mutations map in the 19% of the SV40 A gene that shares no homology with the A gene of polyoma virus. The mutations exist in two different genetic backgrounds: the original set of mutants (dl2401 through dl2405) was prepared, using as a parent SV40 mutant dl862, which has a deletion at the single HpaII site (0.725 map unit). A second set (dl2491 through dl2495) contains the same deletions in a wild-type SV40 (strain SV-S) background. Relative to wild-type SV40, the original mutants showed reduced rates of growth, lower yields of progeny virus and viral DNA, and smaller plaque size; in these properties the mutants resembled parental dl862, although mutant progeny yields were usually lower than yields of dl862, suggesting a possible interaction between the two deletions. The second set of mutants had growth properties and progeny yields similar to those of wild-type SV40; however, Southern blotting experiments indicated that viral DNA replication proceeds at a slightly reduced rate. All of the mutants transformed mouse NIH/3T3 cells and mouse embryo fibroblasts at the same frequency as wild-type SV40. Mutants dl2402, dl2492, and dl2405 consistently produced denser and larger foci in both types of cells. All mutants directed the synthesis of shortened large T antigens. Adenovirus helper function was retained by all mutants. PMID- 6287031 TI - New physical map of bacteriophage T5 DNA. AB - The locations of 103 cleavage sites, produced by 13 restriction endonucleases, were mapped on the DNA of bacteriophage T5. Single- and double-digest fragment sizes were determined by agarose gel electrophoresis, using restriction fragments of phi X174 DNA and lambda DNA as molecular weight standards. Map coordinates were determined by a computer-based least-squares procedures (J. Schroeder and F. Blattner, Gene [Amst] 4:167-174, 1978). The fragment sizes predicted by the final map are all within 2% of the measured values. Based on this analysis, T5st(+) DNA contains 121,300 base pairs (Mr, 80.3 X 10(6) and has a terminal repetition of 10,160 base pairs (Mr, 6.7 X 10(6)). Restriction endonuclease analysis after treatment with exonuclease III and a single-strand-specific endonuclease allowed precise localization of five of the natural single-chain interruptions in T5 DNA. Revised locations for several T5 deletions were also determined. PMID- 6287030 TI - Analysis by microinjection of the biological effects of site-directed mutagenesis in cloned avian leukosis viral DNAs. AB - Cloned avian leukosis viral DNAs were mutagenized in the long terminal repeat, in the leader sequence for env mRNA, and at the poly-env junction. The effect of these mutations in the viral DNA upon its ability to direct virus production or env mRNA synthesis was analyzed by microinjecting the mutant DNAs into chicken embryo fibroblasts and into chicken cells transformed by the env-deficient Bryan strain of Rous sarcoma virus, respectively. The results indicated that: (i) addition of up to 8 base pairs 19 nucleotides upstream of the Hogness box did not block transcription; (ii) deletion of 26 base pairs, including the tRNA primer binding site, allowed synthesis of all viral products and participation in recombination, but replication was blocked; (iii) deletion of fewer than 50 base pairs 250 bases downstream of the long terminal repeat depressed expression of all viral genes; and (iv) deletion of most of the gag and pol genes did not inhibit env mRNA synthesis, but virion packaging of the unspliced transcript was inefficient. PMID- 6287032 TI - Structure and expression of class II defective herpes simplex virus genomes encoding infected cell polypeptide number 8. AB - Defective genomes present in serially passaged virus stocks derived from the tsLB2 mutant of herpes simplex virus type 1 were found to consist of repeat units in which sequences from the U(L) region, within map coordinates 0.356 and 0.429 of standard herpes simplex virus DNA, were covalently linked to sequences from the end of the S component. The major defective genome species consisted of repeat units which were 4.9 x 10(6) in molecular weight and contained a specific deletion within the U(L) segment. These tsLB2 defective genomes were stable through more than 35 sequential virus passages. The ratios of defective virus genomes to helper virus genomes present in different passages fluctuated in synchrony with the capacity of the passages to interfere with standard virus replication. Cells infected with passages enriched for defective genomes overproduced the infected cell polypeptide number 8, which had previously been mapped within the U(L) sequences present in the tsLB2 defective genomes. In contrast, the synthesis of most other infected cell polypeptides was delayed and reduced. The abundant synthesis of infected cell polypeptide number 8 followed the beta regulatory pattern, as evident from kinetic studies and from experiments in which cycloheximide, canavanine, and phosphonoacetate were used. However, in contrast to many beta (early) and gamma (late) viral polypeptides, the synthesis of infected cell polypeptide number 8 was only minimally reduced when cells infected with serially passaged tsLB2 were incubated at 39 degrees C. The tsLB2 mutation had previously been mapped within the domains of the gene encoding infected cell polypeptide number 4, the function of which was shown to be required for beta and gamma viral gene expression. It is thus possible that the tsLB2 mutation affects the synthesis of only a subset of the beta and gamma viral polypeptides. An additional polypeptide, 74.5 x 10(3) in molecular weight, was abundantly produced in cells infected with a number of tsLB2 passages. This polypeptide was most likely expressed from truncated gene templates within the most abundant, deleted repeats of tsLB2 defective virus DNA. PMID- 6287033 TI - Herpes simplex virus mRNA species mapping in EcoRI fragment I. AB - We described the detailed characterization and high-resolution mapping of nine herpes simplex virus type 1 mRNAs encoded in EcoRI fragment I. Four of these mRNAs are partially colinear and encode the same sized polypeptide in vitro. Nucleotide sequence analysis of the DNA around the 5' ends of these mRNAs suggested that the larger may encode a small (ca. 100-dalton) polypeptide not resolvable by in vitro translation. PMID- 6287034 TI - Identification of a nonvirion protein of Aleutian disease virus: mink with Aleutian disease have antibody to both virion and nonvirion proteins. AB - We studied Aleutian disease virus polypeptides in Crandall feline kidney (CRFK) cells. When CRFK cells labeled with [35S]methionine at 60 h postinfection were studied by immunoprecipitation with sera from infected mink, the major Aleutian disease virus virion polypeptides (p85 and p75) were consistently identified, as was a 71,000-dalton nonvirion protein (p71). The peptide maps of p85 and p75 were similar, but the map of p71 was different. p85, p75, and p71 were all precipitated by sera from Aleutian disease virus-infected mink, including those with signs of progressive disease, but heterologous sera raised against purified Aleutian disease virus did not precipitate the nonvirion p71. These results indicated that the nonvirion p71 was unrelated to p85 and p75 and further suggested that mink infected with Aleutian disease virus develop antibody to nonvirion, as well as structural, viral proteins. PMID- 6287036 TI - Variation in the number of copies and in the genomic organization of ecotropic murine leukemia virus proviral sequences in sublines of AKR mice. AB - DNAs isolated from individual mice of four AKR sublines (AKR/J, AKR/N, AKR/Cum, and AKR/Boy) were examined by hybridization of electrophoretically separated restriction enzyme fragments to a 500-base pair, 32P-labeled probe specific for env sequences of ecotropic murine leukemia virus. Variation in the number of proviral DNA copies and in their genomic organization, as reflected by the location of restriction enzyme sites in flanking cellular sequences, was observed both between and within AKR sublines. Evidence is presented for the continual acquisition of new proviruses in the four sublines studied. The ecotropic proviral DNA copies present in the four AKR sublines can be related to their genealogy; each subline contains two or three copies of proviral DNA in common with other sublines and from one to six unique ecotropic proviruses. Overall, a new copy appears about every 12 generations of inbreeding. Some of the unique proviral DNA copies contain internal alterations, as reflected by restriction enzyme maps that differ from those of prototype ecotropic proviruses. PMID- 6287035 TI - Requirements for excision and amplification of integrated viral DNA molecules in polyoma virus-transformed cells. AB - The integration of polyoma virus DNA into the genome of transformed rat cells generally takes place in a tandem head-to-tail arrangement. A functional viral large tumor antigen (T-Ag) renders this structure unstable, as manifested by free DNA production and excision or amplification of the integrated viral DNA. All of these phenomena involve the mobilization of precise genomic "units," suggesting that they result from intramolecular homologous recombination events occurring in the repeated viral DNA sequences within the integrated structures. We studied polyoma ts-a-transformed rat cell lines, which produced large T-Ag but contained less than a single copy of integrated viral DNA. In all of these lines, reversion to a normal phenotype (indicative of excision) was extremely low and independent of the presence of a functional large T-Ag. The revertants were either phenotypic or had undergone variable rearrangements of the integrated sequences that seemed to involve flanking host DNA. In two of these cell lines (ts-a 4A and ts-a 3B), we could not detect any evidence of amplification even after 2 months of propagation under conditions permissive for large T-Ag. An amplification event was detected in a small subpopulation of the ts-a R5-1 line after 2 months of growth at 33 degrees C. This involved a DNA fragment of 5.1 kilobases, consisting of the left portion of the viral insertion and about 2.5 kilobases of adjacent host DNA sequences. None of these lines spontaneously produced free viral DNA, but after fusion with 3T3 mouse fibroblasts, R5-1 and 4A produced a low level of heterogeneous free DNA molecules, which contained both viral and flanking host DNA. In contrast, the ts-a 9 cell line, whose viral insertion consists of a partial tandem of approximately 1.2 viral genomes, underwent a high rate of excision or amplification when propagated at temperatures permissive for large T Ag function. These results indicate that the high rate of excision and amplification of integrated viral genomes observed in polyoma-transformed rat cells requires the presence of regions of homology (i.e., repeats) in the integrated viral sequences. Therefore, these events occur via homologous intramolecular recombination, which is promoted directly or indirectly by the large viral T-Ag. PMID- 6287037 TI - Monoclonal antibodies against baboon endogenous virus and against host cell antigens. AB - Monoclonal antibodies were produced by murine hybridomas after immunization with semipurified baboon endogenous virus. In a solid-phase radioimmunoassay, two antibodies (F12-9 and B9-18) reacted with viral antigen only. The antibodies A6-8 and C9-12 also reacted with virus-producing cells but not with control cells, whereas antibodies E4-6 and D12-2 bound to virus-free cells as well. The cytofluorometry technique confirmed these results and showed a competition between antibodies A6-8 and C9-12 for binding to virus-producing cells as well as a competition between antibodies D12-2 and E4-6 for binding to virus-free human cells. An immune precipitation assay with disrupted virions indicated that antibodies A6-8, B9-18, and C9-12 were directed against the gp70 glycoprotein, and that antibody F12-9 reacted with a viral antigen with a molecular weight of 18,000. The syncytia induced in RSa cells by baboon molecular weight of 18,000. The syncytia induced in RSa cells by baboon endogenous virus could be inhibited either when antibody A6-8 or C9-12 was combined to the virus or when the RSa cells were treated with the anticellular antibody D12-2 or E4-6. These two effects were not observed with Mason-Pfizer virus. Thus, of three antibodies with specificities for viral gp70, two (A6-8 and C9-12) were directed at viral sites responsible for syncytium formation. Another antiviral antibody (F12-9) reacted with a protein of unknown function with a molecular weight of 18,000. The two anticellular antibodies were directed at similar or neighboring epitopes, which may be situated within the receptor to the virus. PMID- 6287038 TI - Adenovirus DNA is associated with the nuclear matrix of infected cells. AB - Viral DNA was found to be tightly associated with the nuclear matrix from HeLa cells lytically infected with human adenovirus type 5. The bound viral DNA, like cell DNA, was resistant to nonionic detergent and to extraction with high-salt (2 M NaCl) solution. However, whereas over 95% of the cell DNA was recovered in the matrix fraction, the amount of associated viral DNA varied during infection. Throughout the lytic cycle, the amount of matrix-associated adenovirus type 5 DNA increased until it reached a plateau level at 20 to 24 h after infection. At this stage, the matrix-bound DNA represented 87% of the total viral DNA; after this stage, additional newly synthesized viral DNA accumulated as non-matrix associated DNA. DNase digestion studies revealed that all viral DNA sequences were equally represented in the matrix-bound DNA both early and late in infection; thus, unlike cell DNA, there seem to be no preferred attachment sites on the viral genome. An enrichment of viral DNA relative to cell DNA was found in the matrix-associated DNA after extensive DNase I digestion. This finding, together with an in situ hybridization study, suggests that the viral DNA is more intimately associated with the nuclear matrix than is cell DNA and probably does not exist in extended loops. PMID- 6287039 TI - Identification and isolation of the main component (gp350-gp220) of Epstein-Barr virus responsible for generating neutralizing antibodies in vivo. AB - The majority of hybridomas we have characterized against Epstein-Barr virions react with the major glycoproteins gp350 and gp220 (gp350/220). One of these antibodies, ID4C-1, neutralizes virus infection in vitro. The presence of gp350/220 on the viral envelope could be confirmed directly by immunoelectron microscopy. We used lectin affinity (ricin) and immunoaffinity (ID4C-1) to purify gp350/220 and show that this material is able to induce potent virus-neutralizing antibodies. Absorption of four human and one rabbit anti-Epstein-Barr virus sera with purified gp350/220 suggests that this is the primary component responsible for generating neutralizing antibodies in vivo. PMID- 6287040 TI - Effects of fowlpox virus infection on lipid metabolism in cultured chicken embryo cells. AB - Lipid biosynthesis was measured in cultured chicken embryo cells after infection with fowlpox virus. Between 24 and 72 h postinfection, fowlpox virus-infected cells incorporated less [14C]acetate and 3H2O into fatty acids and sterols than did mock-infected cells, demonstrating a virus-dependent inhibition of general lipid metabolism. Two specific effects of fowlpox virus infection were an accumulation of C-4 alkylated sterol intermediates and inhibition of monounsaturated fatty acid biosynthesis. PMID- 6287041 TI - Cross-sectional imaging of multilocular cystic nephroma. PMID- 6287042 TI - Re: defining renal anatomy and function with 99m Technetium dimercaptosuccinic acid: clinical and renographic correlation. PMID- 6287043 TI - Ultrastructure of vesicourethral innervation. I. Neuroeffector and cell junctions in the male internal sphincter. AB - The ultrastructure of neuroeffector and cell junctions in smooth muscle of the internal sphincter was studied in the male cat and rat. Muscle cells of the sphincter have the same features of smooth muscle elsewhere, with frequent adherens-type junctions. Neuroeffector junctions are established with sphincter muscle cells by cholinergic and/or adrenergic axons, which probably are not distributed on a 1:1 nerve:muscle basis. The neuroeffector junctions are classified according to their cleft width, their overall morphology, the functional class of their axonal component, and the number of axonal and muscle cell elements involved. These observations unequivocally confirm the principle of dual cholinergic-adrenergic control of the male internal sphincter, and indicate that its mechanism of neuromuscular transmission is both direct via neuroeffector and indirect via muscle cell junctions. PMID- 6287044 TI - Active sodium transport across the epithelium of the human kidney pelvis. Part 1. AB - To clarify whether the epithelium of the human urinary tract possesses an active transepithelial sodium transport, the unidirectional sodium fluxes and the electrical parameters of the epithelium were determined on isolated pieces of renal pelvis. The epithelium was incubated in isotonic Ringer's solution at 37.3C in Ussing-type chambers. The transepithelial potential difference was found to be 15.8 +/- 3.0 mV and the transepithelial electrical resistance 760 +/- 48 omega . cm.-2. The transepithelial, unidirectional sodium fluxes were measured under short-circuit conditions. From both the unidirectional sodium influx from the luminal to the serosal side (7.01 +/- 2.17) and from the efflux in the opposite direction (0.45 +/- 0.07) a transepithelial sodium net flux of 6.56 +/- 2.20 nmol. . minutes-1. cm.-2 was calculated. Since there are no passive forces across the epithelium under short circuited conditions to explain the net flux, an active sodium transport has to be postulated. In all experiments, the simultaneously measured short circuit current (6.43 +/- 2.18 nmol. . minutes-1 . cm-2) was not significantly different from the net sodium flux, and can therefore be taken as a measure of the net flux. PMID- 6287045 TI - A clinical study of 99mTechnetium dimercaptosuccinic acid uptake in obstructed kidneys: comparison with the creatinine clearance. AB - We studied 17 hydronephrotic kidneys owing to stenosis of the pyeloureteral junction. Preoperative uptake of 99mtechnetium dimercaptosuccinic acid was compared to early postoperative unilateral creatinine clearance, measured by urine collection from the nephrostomy tube. An excellent correlation was found. Therefore, uptake of 99mtechnetium dimercaptosuccinic acid represents a reliable parameter of renal function even in the presence of severe urinary tract obstruction. However, 99mtechnetium dimercaptosuccinic acid uptake should be measured more than 24 hours after injection of the tracer. PMID- 6287046 TI - Testicular rupture in blunt scrotal trauma: review of 15 cases with recent application of testicular scanning. PMID- 6287047 TI - Intraluminal renal vein thrombus secondary to metastatic disease to the kidney: a case report. PMID- 6287048 TI - Relapse of gram-negative bacillary meningitis after cefotaxime therapy. PMID- 6287049 TI - Medicine in the USA: historical vignettes. III. Medical sects and their influence. PMID- 6287050 TI - Successful arterial embolization of an insulinoma. AB - A 38-year-old woman had fasting hypoglycemia, hyperinsulinemia, and an arteriographically demonstrable insulinoma. One year after surgical resection, however, hypoglycemia recurred and an arteriogram demonstrated a recurrent intrapancreatic vascular mass. Because of anticipated complications of another surgical approach, we elected to embolize the tumor arteriographically, using microfibrillar collagen. The patient's fasting blood glucose and insulin levels improved rapidly. Eleven months after embolization, the patient remains well, with normal blood glucose levels. PMID- 6287051 TI - Cytomegalovirus vaccine work progressing. PMID- 6287052 TI - Diagnosis of Herpes simplex virus encephalitis. PMID- 6287053 TI - Mechanisms of gene evolution. PMID- 6287054 TI - [Update of pain pharmacology (author's transl)]. PMID- 6287055 TI - [Plasma beta-endorphin and beta-lipotropin levels in patients with trigeminal neuralgia (author's transl)]. PMID- 6287057 TI - [The evaluation and the results of cooperative cholesterol and triglyceride standardization program by WHO-CDC (author's transl)]. PMID- 6287056 TI - [Histopathological and differential diagnosis of testicular tumors, with special reference to WHO's classification (author's transl)]. PMID- 6287058 TI - [Clinical usefulness of acute myocardial infarction imaging by 99mTc pyrophosphate (author's transl)]. PMID- 6287060 TI - [Studies on the characteristic changes in human fibroblasts induced by the proliferation of gastric cancer cells in vitro (author's transl)]. PMID- 6287059 TI - [A case of nodular hidradenoma and freeze-fracture study (author's transl)]. PMID- 6287061 TI - [A case of scirrhous carcinoma of the colon which contained a markedly large amount of glycosaminoglycans (author's transl)]. PMID- 6287062 TI - [An autopsy case of gastric carcinoma, followed by acute gastric dilatation after a biopsy, with past history of gastroparesis diabeticorum (author's transl)]. PMID- 6287063 TI - Enterovirus type 70-neutralizing IgM in animal sera. AB - The neutralizing activity against human enterovirus type 70 was found in serum samples from normal cattle, horses, sheep, goats, swine and chickens raised in Japan. The frequency was variable depending upon animal species and the year of bleeding. The neutralizing activity in bovine sera was shown to reside in IgM by sucrose gradient centrifugation and immune gel electrophoresis. These findings suggested that the neutralizing substance in domestic animal sera is the antibody of IgM class elicited by unidentified viruses antigenically related to human enterovirus type 70. PMID- 6287064 TI - Immunofluorescence for detection of viral antigen in mice infected with Sendai virus. AB - Sendai virus infection transmitted by contact from cagemates was followed by virus titration and immunofluorescence. The virus grew in the respiratory tract and caused macroscopic lesions in all contact mice. The virus grew to a higher titer in the lung than in the trachea. Tracheal smears, however, were found to be the most suitable for the diagnosis of Sendai virus infection by immunofluorescence, since they contained a large number of cells with intense fluorescence. Diagnosis of Sendai virus infection was made by immunofluorescence within a few hours after autopsy made at early stages of infection. PMID- 6287065 TI - [Clinical significance of Tc99m colloid liver angiography for the diagnosis of hepatocellular carcinoma (author's transl)]. PMID- 6287066 TI - Effect of experimental herpes simplex virus vaccine on established ganglionic latency. AB - Mice were inoculated with herpes simplex virus type 1 by the corneal route beforehand. Ten weeks after the inoculation, one group of mice was repeatedly treated with infectious virus-free, detergent-soluble extract of virus-infected cells (DSE) by the intraperitoneal route and the other group served as the controls. The virus recovery rate from the trigeminal ganglia of DES-treated mice was markedly reduced as compared with that of untreated mice, when a small amount of inocula was used. However, no significant decrease in the virus recovery rate was obtained when a large amount of inocula was used, suggesting that treatment with DSE had a limited effect on established ganglionic latency. The level of neutralizing antibody titers in the serum did not seem to be correlated with the efficacy of DSE in mice infected with a small amount of inocula. PMID- 6287067 TI - Responses of salivatory neurons in the medulla oblongata of rabbit. PMID- 6287068 TI - Absorption loss of peptides to the plastic tube in radioreceptor assay of calcitonin and beta-endorphin: protection by detergents. AB - During development of the calcitonin binding assay using rat brain particulate fraction, it was noticed that 125l-salmon calcitonin-l (125-SCT) was partly absorbed by the polypropylene tube surface in an irreversible manner. Absorption was partially prevented by cold SCT and complete prevention was achieved by bacitracin. Triton X-100, Brij 36T, and some Zwittergents in such concentration ranges which appeared not to disrupt the 125l-SCT binding ability of brain tissue. The brain fraction itself exhibited a similar preventive effect. The anti absorbing effect of Brij 36T was also observed in an opiate receptor binding assay for beta-endorphin. These results led us to recommend that the binding assay for these peptides should be done only in the presence of an appropriate anti-absorbant. PMID- 6287069 TI - Effects of estrogen and progesterone on adrenoceptors and cyclic nucleotides in rat uterus. AB - The effects of estrogen and progesterone on adrenoceptors and cyclic nucleotides were studied in ovariectomized rat uterus. The effects on adrenoceptors were examined by measuring mechanical responses to noradrenaline and by binding site determinations with [3H]-dihydroergocryptine and [3H]-dihydroalprenolol. After acute administration of estradiol-17 beta, uterine cyclic GMP was progressively and significantly elevated, but cyclic AMP declined through it was not significant. Phentolamine suppressed this cyclic GMP elevation. Both acute and chronic treatments with estrogen increased the number of alpha-receptors. The increase in uterine cyclic GMP was related to the increase in alpha-stimulation as a result of increased alpha-receptors. Chronic treatment with estrogen increased the number of alpha-receptors. The alpha-effect induced by noradrenaline was bidirectional in the uterus treated with estrogen chronically; there was contraction in normal Tyrode's solution and relaxation in high K+ Tyrode's solution. In other hormonal group, either alpha- or beta-effects produced relaxation. Enhancement of the alpha- and beta-effects by estrogen is attributed to an increased number of the receptors. Acute treatment with estrogen decreased the responses of cyclic AMP to isoproterenol and mechanical reactivity. Progesterone also enhanced the beta-effect as a result of an increase in the number of beta-receptors. PMID- 6287070 TI - [Experience with reconstruction of the carina. Report of two cases (author's transl)]. PMID- 6287072 TI - A pathologic study on initial lesions of enzootic bovine leukosis. PMID- 6287071 TI - Recovery of virus from feces and tissues of chickens infected with cell-culture adapted infectious bursal disease virus. PMID- 6287073 TI - Isolation and properties of adenovirus from canine respiratory tract. PMID- 6287074 TI - Possible association of radioprotective and chemoprotective aminophosphorothioate drug activity with polyamine oxidase susceptibility. AB - The aminophosphorothioate drug S-2-(3-aminopropylamino)ethylphosphorothioate (WR 2721) is both radioprotective and chemoprotective, with potential clinical use in cancer therapies. These distinct properties may be associated with its catabolism by polyamine oxidase (EC 1.4.3.4), which is found in varying amounts in different body tissues. Unfortunately, the metabolite is probably a cytotoxic aldehyde, but it should be detoxified in vivo following adduction with tissue sulfhydryls. Whether conversion and adduction benefit or hinder pharmacologic activity is not known, inasmuch as aldehydes may reduce oxygen-dependent free radicals generated by irradiation of tissues. S-2-(3-aminopropylamino)propylphosphorothioate was similarly a substrate for polyamine oxidase, and the product was cytotoxic. Other aminophosphorothioates [S-2-(4-aminobutaneamino)ethylphosphorothioate, S-2-(5 aminopentylamino)ethylphosphorothioate, and S-2-(4 aminobutaneamino)propylphosphorothioate] were poor substrates and less radioprotective. PMID- 6287075 TI - Search for evidence of feline leukemia virus infection in humans with leukemias, lymphomas, or soft tissue sarcomas. AB - Two-hundred and thirty-nine patients, including 66 with soft tissue sarcomas and 173 with hematologic cancers, were investigated for evidence of infection with feline leukemia virus (FeLV). Included in this group were 2 patients with acute lymphocytic leukemia who had lived in the same household with leukemic cats. None of the patients demonstrated measurable levels of neutralizing antibody to FeLV or had detectable FeLV antigens in peripheral blood normal leukocytes or leukemic cells. In a smaller number of patients, there was no evidence of antibody to FOCMA or of FOCMA on tumor cells in the bone marrow. PMID- 6287076 TI - Isolation and partial characterization of surface components of cell line MDA-MB 231 derived from a human metastatic breast carcinoma. AB - Neuraminidase (Vibrio cholerae) treatment of human metastatic mammary carcinoma MDA-MB-231 cells grown in culture released 0.60-0.63 mg of N-acetylneuraminic acid from 10(9) cells. Incubation of intact cells with a modified trypsin and fractionation by gel filtration gave mainly O-glycopeptides. The presence of O glycosyl-linked chains having one or two carbohydrate residues was confirmed by treatment of the glycopeptide fractions with galactose oxidase, followed by reduction with alkaline sodium borotritide and fractionation. The major glycopeptide fraction, which consisted of 53% carbohydrate and 47% protein, and a minor glycopeptide fraction each inhibited hemagglutination by peanut lectin. These results suggest the presence of O-beta-D-galactopyranosyl-(1 leads to 3)-O (2-acetamido-2-deoxy-alpha-D-galactopyranosyl)-(1 leads to 3)-L-serine (threonine) residues. The absorptive capacities for anti-HLA-A2 and anti-HLA-B8 antisera were slightly greater for intact than for lyophilized cells, which suggested that masking of these major histocompatibility antigens did not occur in intact cells. PMID- 6287077 TI - Effect of type C viral expression and cellular karyotype on human B-lymphoblast tumorigenicity in nude mice. AB - Cultured human hematopoietic cells from several normal and leukemic sources, including those cells initiated after exposure to primate type C retroviruses were tested for their capacity to induce tumors in young athymic BALB/c (nu/nu) mice after sc inoculation. An attempt was made to correlate these results with virus expression and chromosome patterns. Progressively growing tumor formation was observed in 5 of 18 normal diploid B-lymphoblast lines from normal peripheral blood and in one of three diploid B-lymphoblast lines from leukemic donors established after infection with primate type C viruses (gibbon ape leukemia virus or simian sarcoma virus). In contrast, none of eight spontaneously transformed B-lymphoblast lines with normal diploid karyotypes formed progressively growing tumors, although one formed a tumor that remained the same size (0.5 cm) for several months. Progressive tumor formation occurred in four of seven previously established cell lines of different cell types that had abnormal karyotypes. Of the normal diploid B-lymphoblast cultures exposed to type C viruses, 12 were tested for the presence of viral RNA and structural proteins (p12, p30, gp70), and this information was correlated with tumorigenicity. Four of the six cultures expressing viral RNA or proteins were tumorigenic, whereas only one of six cultures that did not express virus information was positive. The results of this study suggest that expression of type C viral RNA and proteins by human B-lymphoblasts increases their tumorigenicity in nude mice. It is also apparent that caution must be used in attempts to correlate cell tumorigenicity and chromosome abnormalities in nude mice. PMID- 6287078 TI - Comparison of ovariectomy and retinyl acetate on the growth of established 7,12 dimethylbenz[a]anthracene-induced mammary tumors in the rat. AB - Prolonged exposure to retinyl acetate (RA) in the diet inhibits the development of 7,12-dimethylbenz[a]anthracene (DMBA)-induced mammary cancers in rats. The effectiveness of RA was examined when given 6 months after the administration of DMBA. Non-inbred female Sprague-Dawley rats with DMBA-induced mammary tumors were divided into 3 groups and treated for 4 weeks as follows: Group 1 served as controls, group 2 was ovariectomized, and group 3 received 328 mg RA/kg diet. Ovariectomy (OVX) markedly reduced both the number and size of the tumors. RA administration failed to induce any significant regression in tumor number but significantly retarded tumor growth when compared to tumor growth in group 1 controls. The levels of estradiol, progestin, and prolactin (PRL) receptors were significantly reduced after OVX, whereas only the levels of PRL receptors declined significantly after RA administration. Circulating progesterone concentrations were not affected in the RA-treated group but the plasma PRL level was significantly increased. The present studies show that if treatment with RA is delayed until 6 months after carcinogen administration, the protective effect of RA can still be observed although its effectiveness is less dramatic than when it is administered earlier. PMID- 6287079 TI - Individually distinct tumor-specific cell surface antigen identified by monoclonal antibody on a Rous sarcoma virus-induced mouse tumor. AB - Hybrid cell lines were prepared by the fusion of BALB/c myeloma P3U-1 cells with the lymphocytes of BALB/c mice that were immunized with syngeneic Rous sarcoma virus (RSV)-induced tumor CSA1M cells. Three clones of the hybrid progeny (3.4B2, 3.4C6, and 3.5C11) produced cytotoxic IgM antibodies against CSA1M cells. One of the clones, 3.5C11, was chosen for analysis of the detailed specificity. Both direct cytotoxicity assays and absorption tests revealed that monoclonal antibody from 3.5C11 was positive only with CSA1M cells and that it failed to react with other tumors, including 20 RSV-induced mouse tumors, and normal cells. The 3.5C11 monoclonal antibody alone, with or without exogenous complement, was suppressive in the therapy of ip injected CSA1M tumor in syngeneic hosts, and significant prolongation in survival was seen in the treated mice. These results clearly showed presence of an individually distinct tumor-specific cell surface antigen on an RSV-induced mouse tumor. PMID- 6287080 TI - Distribution of hepatitis B surface and core antigens in human liver cell carcinoma and surrounding nontumorous liver. AB - The distribution of hepatitis B surface antigen (HBsAg) and core antigen (HBcAg) in surgically removed liver cell carcinomas of 60 Japanese patients was studied by an immunohistochemical method. HBsAg was localized in the cytoplasm, and HBcAg was mainly in the nuclei of normal-appearing liver cells, dysplastic liver cells, and tumor cells. In some instances both HBsAg and HBcAg were contained in the tumor cells and liver cells. HBsAg was found in the nontumorous liver tissue of 30 patients (50%); nine of the HBsAg-containing nontumorous livers also contained HBcAg. HBsAg and/or HBcAg was detected in tumor cells in 7 (23%) of 30 cases with HBsAg-positive liver diseases. PMID- 6287081 TI - Elevated dTTPase activities in sera of patients with advanced cancer. PMID- 6287082 TI - Persistence of transforming and nontransforming Epstein-Barr virus in high passages of P3HR-1 cell lines. AB - At least three laboratories have reported that the P3HR-1 line, which had originally produced transforming Epstein-Barr virus (EBV), now produces only the nontransforming variant. Studies to determine whether these findings were universal or a consequence of specific cell lines or culture conditions were undertaken in P3HR-1 cultures of identical HLA types from five sources. All of the EBV preparations derived from cell lines cultured at 32, 34, and 35 degrees C transformed cord blood lymphocytes, whereas virus propagated at 37 degrees C did not usually transform. Furthermore, indirect immunofluorescence revealed that a monoclonal antibody directed against transforming EBV membrane glycoprotein bound to 10-12% of the P3HR-1 cells that had been continuously propagated at 34 degrees C, but the antibody did not bind to the same cells cultured at 37 degrees C. Although virus expression was completely repressed in transformed cord blood cells, transforming virus could be rescued by superinfection with nontransforming P3HR-1 EBV. Cells transformed with P3HR-1 virus induced poorly differentiated lymphomas in athymic nude mice after seven or eight passages. Whether all P3HR-1 cells have the potential to produce detectable quantities of transforming virus remains to be determined. PMID- 6287083 TI - A high-tumor-incidence subline of the D1 mouse mammary hyperplastic outgrowth line: effect of carcinogens. AB - Mammary tumorigenesis in some mouse strains is characterized by the appearance of a preneoplastic lesion, the hyperplastic alveolar nodule (HAN). The biology of the HAN has been characterized primarily through the study of stable outgrowth lines of serially transplanted HAN. One outgrowth line, Dl, which was developed and carried in female BALB/c mice, has been described as a low-tumor-incidence line that does not express murine mammary tumor virus (MuMTV) and is susceptible to hormonal, chemical, and viral carcinogens. In this report, a high-tumor incidence subline of Dl, Dl/UCD, is described. Although Dl/UCD, like Dl, is susceptible to the chemical carcinogen 7,12-dimethylbenz[a]anthracene, an increase in tumor incidence was not observed when Dl/UCD outgrowth was exposed to hormones by means of pituitary isografts. Unlike Dl, Dl/UCD is refractory to the carcinogenic action of MuMTV. Both Dl and the Dl/UCD subline contained the endogenous MuMTV provirus but did not contain exogenous MuMTV provirus sequences. MuMTV antigen was not detected in Dl/UCD outgrowths or tumors. RNA hybridizable to MuMTV complementary DNA was detected in some Dl/UCD outgrowths and tumors but did not appear to correlate with tumorigenesis. PMID- 6287084 TI - In vitro infection of lymphocytes with Marek's disease virus. AB - Suspension cultures of splenic lymphocytes, incubated at 41 degrees C, became infected with Marek's disease virus (MDV) following exposure to a) other infected lymphocytes, b) infected chicken kidney monolayer cultures, or c) cell-free MDV. Both viral antigen expression and virus isolation could be demonstrated after more than 40 passages made by the addition of fresh spleen cells at 2- to 3-day intervals. Susceptibility of spleen cells from bursectomized chickens was markedly lower than that of cells from intact birds. Furthermore, when spleen cell suspensions were depleted of cells having characteristics of bursa-derived cells, e.g., those with surface IgM, Fc receptors, or ability to adhere to nylon wool, the susceptibility of the cell suspension was diminished. Enrichment of the suspension with cells having those features enhanced overall susceptibility. The target cells for virus infection in vitro also were shown to be nonphagocytic, to be of low or medium density, and to bear Ia-like antigen. In vitro susceptibility to infection of spleen cells did not correlate with the genetic susceptibility of the donor to Marek's disease. PMID- 6287085 TI - Pseudohypoparathyroidism. AB - The current understanding of the adenylate cyclase complex has permitted us to regard PHP has a heterogeneous group of disorders related to each other through the common feature of resistance to PTH. Patients may present with hormone resistance limited to PTH, or with a metabolic disorder attributable to the resistance of multiple tissues to hormones that activate adenylate cyclase. The majority of the latter patients show deficient G-unit activity. The clinical expression of multiple hormone resistance in patients with the G-unit defect appears to be rather variable, despite an identical quantitative defect; hormone resistance seems incomplete, and patients may be hypocalcemic despite the presence of hyperparathyroid bone disease. It is therefore likely that the detailed biochemical study of patients with PHP may ultimately reveal a multiplicity of abnormalities in the hormone-receptor adenylate cyclase system that will explain the variability of hormone resistance. PMID- 6287086 TI - [Role of catecholamine in activating glycogenolysis in experimental myocardial infarct complicated by ventricular fibrillation]. AB - The experiments on anaesthetized dogs have shown that with artificial ventilation and open chest adrenalin in arrhythmogenic and nonarrythmogenic doses enhances glycogenolysis in the heart proportionally to the dose used Block of beta adrenergic receptors by propranolol decreased the activation of glycogenolysis in the ischaemic zone and the frequency of ventricular fibrillation after occlusion of the coronary artery. Exhaustion of noradrenaline reserves by reserpin does not influence the intensity of glycogenolysis and the frequency of fibrillation in the acute period of myocardial infarction. It is believed that the activation of glycogenolysis in experimental myocardial infarction is related to the action of adrenalin. PMID- 6287088 TI - [Hemorrhaging chemodectoma of the stomach]. PMID- 6287087 TI - [Disseminated intravascular coagulation and circulatory shock]. AB - The authors describe the pathophysiology of disseminated intravascular coagulation and the coagulopathy of utilization. The clinical and laboratory data on different types of shock are described. The efficacy of different antithrombotic agents in shock with disseminated intravascular coagulation is shown. PMID- 6287089 TI - [Congenital solitary adenoma of the liver transformed into hepatocellular cancer]. PMID- 6287091 TI - Impairment of peripheral nerve in vibrating tool operators examined by nerve conduction velocity. PMID- 6287090 TI - [Superoxide-dismutase and superoxide-radical-release in rheumatoid arthritis (author's transl)]. AB - Polymorphonuclear leukocytes (PMNs) release superoxide anion (O-2) when they are exposed to a phagocytic stimulus. Intracellularly the copper-containing enzyme superoxide dismutase (SOD) protects against the toxic effects of O-2. To investigate the role of O-2 and SOD in the inflammatory process we determined the O-2 release and SOD content in PMNs. In PMNs of children with rheumatoid arthritis the SOD activity was diminished compared to healthy controls. Upon stimulation with opsonized zymosan PMNs obtained from children with rheumatoid arthritis generated greater amounts of superoxide anion than control cells. The "SOD deficiency" in PMNs of children with rheumatoid arthritis may promote this extreme release of the toxic superoxide radical inducing the damage of the connective tissue. The involvement of superoxide dismutase and superoxide anion in inflammatory process may induce further studies, leading hopefully to an appropriate understanding or even to new principles in the treatment of the rheumatoid arthritis. PMID- 6287092 TI - Benzodiazepines and the gamma-aminobutyric acid (GABA) receptor. PMID- 6287093 TI - [Psychiatric nursing. A holiday camp with psychiatric patients]. PMID- 6287095 TI - [Terres des Hommes Switzerland: basic medicine in Brazil]. PMID- 6287094 TI - [Outpatient nursing care: training in rehabilitation. Mrs. W. becomes independent again]. PMID- 6287096 TI - [Position of special interest student groups of Canton Bern on the structure of the educational system in the nursing professions]. PMID- 6287097 TI - [Rights and duties of students]. PMID- 6287098 TI - [Single girls on the road]. PMID- 6287099 TI - [The nursing professional in search of an identity. Interview by Nicole-France Exchaquet]. PMID- 6287101 TI - [Physicians without frontiers]. PMID- 6287100 TI - [Can one choose one's death?]. PMID- 6287102 TI - [Experience of a Swiss nurse in Africa]. PMID- 6287103 TI - [Amnesty International. Violation of human rights in Morocco]. PMID- 6287104 TI - Biochemical analyses of EDTA extracts and collagenase digests from bone and skin of Wistar rats. AB - Using EDTA extraction and collagenase digestion, rat bone and rat skin were compared in terms of their content of hydroxyproline, hexoses, uronic aicd, sialic acid and plasma proteins. The collagen content of the organic matrix from both tissues was similar. Greater differences were observed in the sialic acid and uronic acid content of the matrix, bone containing higher amounts; smaller differences were found in the levels of hexoses, albumin, IgG and alpha 1 acid glycoprotein, which are higher in EDTA extracts from bone. The DEAE-cellulose chromatography of the EDTA extracts and soluble collagenase digests indicated the presence of a variety of glycoproteins and a proteoglycan fraction. An acidic glycoprotein, corresponding to sialoprotein, was present in bone but not in skin extracts. PMID- 6287105 TI - [Steroid ligands of androgen binding protein. The enantiomers of trans-trans and cis-trans perhydrohexestrols and trans-trans perhydrodiketones]. AB - In the framework of a study on nonsteroidal androgens, the authors previously observed that in perhydrohexestrol series, the (+/-)-[(cis-4 hydroxycyclohexyl) 4(trans-4 hydroxycyclohexyl)-hexane] or cis-trans perhydrohexestrol and especially the (+/-)-(3,4 bis (trans-4-oxocyclohexyl)-hexane) or trans-trans perhydrodiketone, there is no affinity for AR (androgen receptor of the prostate) but they bind with high and specific affinity to the testosterone binding sites on ABP (epididymal androgen-binding protein). In this work, we describe the preparation, the stereochemistry and biological activities of trans-trans and cis trans perhydrohexestrols and of trans-trans perhydrodiketone as well as their corresponding enantiomers. Biologically the tests AR and ABP are negative for the trans-trans perhydrohexestrol and of trans-trans perhydrodiketone and for its enantiomers. However for the enantiomers of cis-trans perhydrohexestrol and of trans-trans perhydrodiketone, the affinity of the (+) enantiomer for ABP is superior to that of the racemic and that of the (-) enantiomer, whereas the affinity for AR are zero. Chemically, the stereochemistry of the three racemics has been established by X-ray crystallographic analysis or by 1H n.m.r. The n.m.r. spectra have been analyzed in terms of chemical shifts and coupling constants. PMID- 6287107 TI - Effects of stimulation on the steroid profile formed by rat adrenal capsule tissue incubated in vitro. AB - A characteristic of the response of non-dispersed rat adrenal capsule tissue (mainly zona glomerulosa) to ACTH stimulation is that corticosterone and aldosterone production is increased whereas 18-hydroxy-deoxycorticosterone (18-OH DOC) is not. The effects of potential second messengers on the steroid profile were compared with those of ACTH and K+ ions in adrenal capsule incubations. ACTH stimulated corticosterone, 18-hydroxycorticosterone and aldosterone, but not 18 OH-DOC. In contrast, K+ (5.9 mM) stimulated 18-OH-DOC as well as the other capsule products. Compared with controls incubated with EDTA, the addition of Ca2+ and Mg2+ ions invariably stimulated aldosterone and 18 hydroxycorticosterone, while the effects on corticosterone were variable, and 18 OH-DOC production was unaltered. Addition of dibutyryl cyclic AMP (1 mM and 0.3 mM) or cyclic GMP (1mM) stimulated all products. The results show that Ca2+ ions and dibutyryl cAMP may have slightly different effects in that the cyclic nucleotides can stimulate all products whereas Ca2+ and Mg2+ ions preferentially support aldosterone and 18-hydroxycorticosterone. However, none of the potential intracellular stimulants studied fully reproduce the characteristic response of non-dispersed tissue to ACTH in which the secretion of corticosterone and 18 hydroxy-DOC is disassociated. PMID- 6287106 TI - Effect of corticosteroid binding proteins on the steroidogenic activity of bovine adrenocortical cell suspensions. AB - The possible role of steroid binding proteins in the hormonal secretion process of a steroidogenic tissue was examined using bovine adrenocortical cell suspensions, either under basal conditions or in the presence of half-maximally active concentration (1 x 10(-9) M) of synthetic adrenocorticotropic hormone (ACTH). Three types of plasma cortisol binding proteins were used, namely bovine serum albumine (BSA), purified transcortin (CBG) and purified anticortisol immunoglobulins (IgG). When added to the incubation medium, CBG (at 1 x 10(-10) to 2 x 10(-9) M cortisol binding sites) and anticortisol IgG (at 4.8 x 10(-10) to 3 x 10(-9) M cortisol binding sites) did not influence either the basal nor the ACTH-stimulated net cortisol production of the cell preparations. Whereas crystallized and delipidated BSA showed also no effect, crude commercial BSA preparation (Cohn fraction V) exhibited an ACTH-like cofactor effect which resulted in a marked increase in the net cortisol production by stimulated cells. These observations might be explained by the presence in crude BSA of lipoprotein cholesterol complexes, possibly acting as an extracellular source of cholesterol available for corticosteroidogenesis. It may be concluded that specific high affinity cortisol binding systems present outside adrenocortical steroidogenic cells do not influence their secretory activity under short term in vitro condition. In addition, it can be stressed that use of ill defined protein preparations (e.g. crude BSA) may lead to artifactual observations in the study of the differentiated functions of isolated steroidogenic cells. PMID- 6287108 TI - Determination of urinary free cortisol and cortisone by sequential thin layer and high-performance liquid chromatography. AB - A specific, rapid and sensitive method for urinary free cortisol and cortisone utilizing sequential thin layer (TLC) and high-performance liquid chromatography (HPLC) was developed. After addition of prednisone as the internal standard to 1 ml of urine, extraction of the steroids was accomplished by automated sorption on and desorption from a styrenedivinylbenzene copolymeric resin cartridge. Further purification was carried out by TLC. Quantitation of the recovered steroids at 254 nm was achieved during HPLC in a reverse phase system with 21-deoxycortisone as the external standard. When the values of urinary free cortisol both from normal subjects and patients were compared with those obtained by a current RIA procedure, the greater specificity of the new method was clearly demonstrated. Simultaneous measurement of both urinary free cortisol and cortisone in various clinical states appears to offer a more complete index of adrenocortical function than urinary free cortisol alone. PMID- 6287109 TI - In vitro effects of progesterone and estradiol-17 beta on choleragen activated Xenopus oocyte adenylate cyclase. AB - The basal cAMP levels of full-grown defoliculated Xenopus oocytes were shown to average 1.2 pmol per oocyte. Follicle cAMP concentration was not significantly different from that of denuded oocyte and was found, when expressed as unit of volume to be independent of follicle size. In vitro gonadotropin treatment of follicle does not affect cAMP accumulation. During the course of progesterone induced maturation we failed to detect any modification of the oocyte cAMP content. Cholera toxin treatment increased cAMP levels; estradiol-17 beta potentialized cholera toxin action and progesterone antagonized it. Estradiol-17 beta was also found to reduce the amount of cholera toxin necessary to inhibit 50% of the progesterone induced meiosis maturation. PMID- 6287110 TI - Ethanol-induced conditioned taste aversions in mice that differ in neural sensitivity to ethanol. PMID- 6287111 TI - Hormone receptor levels related to histological parameters of tumor-host relationships in female breast carcinoma. AB - A series of 115 female breast carcinomas was studied with special emphasis on the relationships between the hormone receptor (ER and PR) levels and the histological parameters reflecting tumor-host reactivity. Carcinomas were classified according to their nuclear grade (NC) and the stromal host reactions; perivenous lymphocyte infiltration (PVI), lymphocyte (LD), plasma cell (PCI), and mast cell infiltration (MI) were assessed. NG did correlate directly with the ER positivity, but not with the absolute ER values. LI and PCI, but not MI and PVI, showed an inverse relation to the ER values, but not to those of PR. ER and to lesser extent the PR values were higher in postmenopausal than in premenopausal women. The findings are discussed in the light of tumor-host relationships, and conclusion is drawn that the receptor determinations used in connection with the histological evaluation of these parameters (NG especially) are of definite benefit in dividing the breast cancer patients into groups of different expectancy of the future outcome of their disease. PMID- 6287112 TI - [Pharmacology of the slow calcium channel]. PMID- 6287113 TI - [Immunopharmacology and pharmacology of diethyldithiocarbamate (DIC)]. PMID- 6287114 TI - Photoaffinity labeling of opioid receptor with morphine-7,8-oxide (morphine epoxide). AB - The opioid receptor mediating inhibitory action of morphine in the electrically stimulated guinea pig ileum was irreversibly photoinactivated by morphine epoxide (3 X 10(-6) M). Morphine epoxide (up to 3 X 10(-5) M) did not influence the responses of rat vas deferens (epsilon-receptor) or rabbit vas deferens (kappa receptor) to electrical stimulation. Effective concentrations of morphine epoxide were much lower in the guinea pig ileum (mu-receptor) than in the mouse vas deference (delta-receptor). The inhibitory action of [Met]-enkephalin on the twitch responses of the rat vas deferens and mouse vas deferens to electrical stimulation were not influenced after irradiation in the presence of morphine epoxide (3 X 10(-6) M). Therefore, morphine epoxide is probably a useful probe for photoaffinity labeling of the mu-receptor in vitro. PMID- 6287115 TI - 11th annual UCLA symposia. Abstracts: Evolution of hormone receptor systems. PMID- 6287118 TI - Some indications for inverse DNA duplication. PMID- 6287116 TI - Influence of selective processes on the amino acid compositions of proteins: collagen, cytochrome c, ferredoxin and alpha-crystallin. PMID- 6287117 TI - Strategies for constructing complementary DNA for cloning. PMID- 6287119 TI - Prognosis of pulmonary scar carcinoma. AB - Eighty-one patients with resectable primary peripheral lung carcinomas were studied to determine the effect of associated scarring on prognosis. Twelve tumors (15%) originated from bronchi. 24 (30%) were associated with scars, and 45 (55%) were not associated with either bronchus or scar (non-scar). Scar carcinomas differed significantly in cell type from bronchogenic and non-scar tumors in that 21 (88%) scar carcinomas were either adenocarcinomas or bronchioloalveolar carcinomas (p less than 0.001). Origin (bronchogenic, scar, non-scar) independent of cell type and tumor stage did not significantly influence survival. Stage of disease independent of cell type or origin affected survival (p less than 0.0001), as did cell type independent of tumor stage or origin (p less than 0.0001). Stage I disease and bronchioloalveolar carcinoma were associated with longer survival, while Stage II or III disease, small cell anaplastic carcinoma, and adenocarcinoma were associated with reduced survival. We conclude that associated scar influences cell type of peripheral lung carcinoma but does not influence patient survival, even among patients with similar cell type and stage of disease. PMID- 6287120 TI - Malignant fibrous histiocytoma of the mediastinum and lung: a report of three cases. AB - One case of primary malignant fibrous histiocytoma of the lung and two cases of that tumor in the mediastinum are reported. Primary malignant fibrous histiocytoma is rare in those areas, appearing more commonly in deep fascia and skeletal muscles of the extremities and torso and in the retroperitoneum. Most of the tumors contain both fibroblast-like and histiocyte-like cells; some contain pleomorphic giant cells and inflammatory cells. They are often confused with other sarcomas, and their true biologic potential is not clearly defined. Radiation appears to be a very useful adjunct to surgical therapy and was used in the cases reported here. PMID- 6287121 TI - Pulmonary chondroma, extra-adrenal paraganglioma, and gastric leiomyosarcoma: Carney's triad. AB - The triad of pulmonary chondroma, extra-adrenal paraganglioma, and gastric leiomyosarcoma has been reported in five patients, all young women. The association of tumors apparently constitutes a syndrome. This paper describes the sixth affected patient, also a young woman, draws attention to the frequency of thoracic paraganglioma in the syndrome, and notes that the pulmonary cartilaginous lesion has frequently been misinterpreted as pulmonary hamartoma. Early exploration of the chest is indicated not only to rule out the possibility of metastatic gastric tumor but also to search for paragangliomas. PMID- 6287122 TI - Hepatic oil embolization following lymphangiography. A report of 12 cases. AB - The authors report on 12 personal observations of lymphoportal fistulas, which represented 0.24% of their series of lymphographies. This rare complication of lymphography has no clinical or biological liver manifestations. Review of 71 cases in the literature confirmed that hepatic oil embolization only occurs when there are lymphatic masses, with or without associated thrombosis of the inferior vena cava. PMID- 6287123 TI - Alterations in the properties of beta-adrenergic receptors of myocardial membranes in aging: impairments in agonist-receptor interactions and guanine nucleotide regulation accompany diminished catecholamine-responsiveness of adenylate cyclase. AB - The effects of age on cardiac beta-adrenergic receptor linked adenylate cyclase system were studied using homogenates and membrane fractions of ventricular myocardium from young (3-4 months' old) and aged (24-25 months' old) rats. The number of beta-adrenergic receptor sites and the basal adenylate cyclase activities were essentially similar in young and aged hearts. On the other hand, striking age-associated alterations were seen in the properties of beta-receptors and adenylate cyclase in cardiac membranes. (A) The beta-receptor affinities for catecholamine agonists were reduced 10- to 20-fold in aged compared to young heart; the receptor affinity for antagonists were unaffected by age. (B) Guanine nucleotides caused a 9- to 17-fold reduction in beta-receptor affinity for isoproterenol in young heart; this guanine nucleotide-promoted reduction in receptor affinity for agonist was virtually absent in aged heart. (C) Guanine nucleotide-dependent stimulation of adenylate cyclase by isoproterenol was only 20-50% in aged heart compared to three-fold in young heart; beta-receptor independent activation of the cyclase by guanine nucleotides and NaF were also reduced similarly in aged heart. These results demonstrate a marked age associated deterioration in the functional integrity of beta-receptor linked adenylate cyclase system in aged heart. Further, the findings identify beta receptor and guanine nucleotide regulatory protein as the components of the cyclase system affected in aging. It is suggested that the failure to form a "high-affinity" agonist-receptor complex, owing to the age-related decrement in receptor affinity for agonists, and the apparent inability of guanine nucleotide regulatory protein to effect appropriate molecular transitions at the level of both beta-receptor and the catalytic unit (i.e. the cyclase enzyme), contribute to the loss of catecholamine- and guanine nucleotide-responsiveness of adenylate cyclase in aged heart. This age-associated abnormality provides a biochemical basis for the well-recognized decline in adrenergic control of aging myocardium. PMID- 6287124 TI - Age-linked changes in the activity of enzymes of the tricarboxylate cycle and lipid oxidation, and of carnitine content, in muscles of the rat. AB - The activities of citrate synthase, NAD-isocitrate dehydrogenase and 2 oxoglutarate dehydrogenase were measured in homogenates of soleus, diaphragm and heart muscles of the rat, in an attempt to define potential tricarboxylate cycle activity and its response to aging. Activities were significantly decreased in 24 month animals versus 6-month controls in every case (except 2-oxoglutarate dehydrogenase in heart muscle). Age-linked decrements were greatest in the soleus and least in heart. Cytochrome oxidase was measured as an index of total respiratory chain activity and decreased significantly in each case, with the smallest decrease in the heart. Acyl-CoA dehydrogenase and 3-hydroxyacyl-Co-A dehydrogenase were measured as an index of beta-oxidative activity; the former decreased in soleus and diaphragm, the latter in soleus and heart, with the decrease in the soleus being the greater. Carnitine acetyl- and palmitoyltransferases were measured, together with the muscle content of carnitine and acylcarnitine, as determining the potential rate of entry of acyl groups into the mitochondria for oxidation. Carnitine acetyltransferase activity was decreased with age in each of the muscles, but to the greatest extent in the heart. Carnitine palmitoyltransferase was decreased in both soleus and diaphragm. Carnitine content was decreased most in the soleus and the heart and to a lesser extent in the diaphragm. It is concluded that there is a generalized decline in oxidative activity in all of these muscles with age, on the basis of wet weight; this occurs to the greatest extent in the soleus and to the least extent in the heart. There is, in addition, a specific deficiency in the ability to oxidize fatty acids, relative to other substrates, in heart muscle. PMID- 6287125 TI - Studies on thymocyte subpopulations in guinea pigs. Differences in cell volume and proliferative ability in vitro of two populations separated by density gradient centrifugation with Percoll. AB - Thymus cells from guinea pigs were separated according to buoyant density by centrifugation with PVP-coated colloidal silica particles (Percoll). By creating an S-shaped density gradient, two populations (referred to as peak-I and peak-II cells) were obtained which differed in size as well as ability to spontaneous proliferation in vitro. Peak I contained low density cells of large size and was highly enriched with DNA-synthesizing cells. These continued to proliferate in culture for at least 30 h as demonstrated by mitotic studies in the intervals 0 10 and 20-30 h. In order to grow in vitro, however, the cycling cells of peak I depended on the medium (RPMI 1640) being supplemented with L-alanine. The high density cells of peak II constituted 70% of the thymocytes and had a small and uniform volume. This population was depleted of proliferating cells. The simple and rapid separation of these two major populations is considered a useful first step for the further characterization of thymocyte subpopulations. We suggest that peak I primarily includes proliferating precursor cells from the cortex as well as mature, immunocompetent cells. Peak II consists largely of small cortical cells. PMID- 6287126 TI - [Treatment of gastro-esophageal reflux using a potable gaviscon suspension]. PMID- 6287127 TI - [A molecule active in deficiency and metabolic neuropathies]. PMID- 6287129 TI - Structure-activity relationships related to ceftizoxime (FK 749). PMID- 6287128 TI - [Standardization of methods for determining bacterial sensitivity to 6/N,N-1',6' hexylene formamidinopenicillanic acid (preparation HX)]. PMID- 6287130 TI - Multiple opioid receptors. PMID- 6287132 TI - Claforan (cefotaxime sodium). PMID- 6287131 TI - Angiotensin-converting enzyme inhibitors: medicinal chemistry and biological actions. AB - A decade of experimentation, with work in the last four years greatly accelerated by the availability of captopril, favors the conclusion that angiotensin converting enzyme inhibitors exert their antihypertensive effect by blocking the formation of angiotensin II. The apparently anomalous efficacy of ACE inhibitors in hypertension characterized by low circulating renin levels may point to the importance of the vascular renin-angiotensin system in controlling blood pressure. Neither kinin potentiation nor blockade of the CNS renin-angiotensin system appear to play a significant role in the antihypertensive effect of captopril. Pharmacological and clinical studies on ACE inhibitors of novel chemical structure will establish their utility as antihypertensive agents and may lead to greater understanding of the mechanism of action of all ACE inhibitors. PMID- 6287133 TI - [Fundamentals of echographic diagnosis of acute and chronic sinusitis (author's transl)]. AB - Changes of sound permeability in facial bones are caused by inflammatory or tumorous reaction in the mucosa of the sinuses; echo-orbitography (8-10 MHz) shows in this case extraorbital echo signals if bony walls are thin enough (e.g. Lamina papyracea, anterior and upper wall of the antrum). Further examinations of facial bones show early inflammatory changes of bony tissue in sinusitis without influencing the echographic pattern; the amount of hydroxylapatite remains unchanged. PMID- 6287134 TI - Suppressive effect of dynorphin-(1-13) on luteinizing hormone release in conscious castrated rats. AB - The effect of intraventricular administration of dynorphin-(1-13) on luteinizing hormone (LH) release was studied in castrated conscious rats. The administration of 5 micrograms of dynorphin-(1-13) into the lateral ventricle inhibited LH secretion. Intravenous administration of naloxone blocked this suppressive effect of dynorphin on LH release. These results suggest a possible role of dynorphin, in addition to beta-endorphin and Met5-enkephalin, in the control of LH release in male rats. PMID- 6287135 TI - The effect of vagotomy on the satiety effects of neuropeptides and naloxone. AB - As abdominal vagotomy blocks the satiety effect of cholecystokinin-octapeptide, we felt it would be worthwhile to examine whether the satiety effect of any of the other putative satiety neuropeptides was mediated through the vagus. We confirmed that the satiety effect of peripherally administered cholecystokinin (10 micrograms/kg) was mediated through the vagus. In addition, the satiety effect of peripherally administered TRH (8 mg/kg) also was not present in vagotomized animals. Vagotomy had no effect on the satiety effects of peripherally administered bombesin, calcitonin and naloxone. Nor did vagotomy alter the satiety effect produced by central administration of bombesin, TRH, calcitonin nor naloxone. PMID- 6287136 TI - Epidermal growth factor and cyclic AMP stimulation of distinct protein kinase activities in Leydig cell tumor membranes. AB - Epidermal growth factor (EGF) and cyclic AMP were found to stimulate distinct protein kinase activities in plasma membranes prepared from the M5480P murine Leydig cell tumor. EGF stimulated the phosphorylation of two protein bands with apparent molecular weights of 60,000 and 180,000, while cyclic AMP stimulated the phosphorylation of a minor component of molecular weight 220,000. The two types of kinases could also be distinguished on the basis of differential susceptibility to conditions of membrane preparation. These results suggest that EGF stimulates a cyclic AMP-independent protein kinase in murine Leydig cell tumors at the level of the plasma membrane. PMID- 6287137 TI - Presynaptic change associated with long-term potentiation in hippocampus. PMID- 6287138 TI - Physiological effects of high dose naloxone administration to normal adults. AB - The intravenous bolus administration of high doses of the opioid receptor antagonist naloxone (in the mg./kg. range) to normal adults produced dose dependent increases in systolic blood pressure and respiratory rate. No significant alterations in other physiological processes were found although there were trends for increases in oral temperature and pulse rate with increasing dose. These results suggest the involvement of an endogenous opioid system in the tonic regulation of human systolic blood pressure and respiration and the probable inadequacy of doses of naloxone previously utilized in human studies to effect complete blockade of endogenous opioid systems. PMID- 6287140 TI - Comparison of muscarinic and beta adrenergic receptors between bovine peripheral lung and tracheal smooth muscles: a striking difference in the receptor concentration. AB - This study was undertaken to compare the activity of muscarinic and beta adrenergic receptors in bovine peripheral lung to the corresponding receptor activity in tracheal smooth muscle. We used [3H] quinuclidinyl benzilate (QNB) and [3H]dihydroalprenolol (DHA) to measure muscarinic and beta receptor activity, respectively. Binding to QNB and DHA at 25 degrees C was rapid, reversible, saturable and of high affinity. The order of potency for cholinergic and adrenergic agents competing for binding was compatible with muscarinic and beta 2 adrenergic potencies. We found that the concentration of muscarinic receptor binding sites was 37-fold greater in the tracheal muscle preparation (2805 +/- 309 fmol/mg protein) than in the peripheral lung preparation (76 +/- 28 fmol/mg protein). Unlike muscarinic receptors, the lung contained 8-fold higher concentration of the beta adrenergic receptors than did the tracheal muscle (1588 +/- 417 vs. 199 +/- 42 fmol/mg protein). The dissociation constant or the agonist's inhibitory constant (Ki) for either receptor binding site, however, was not significantly different between the two tissues. Furthermore, in vitro contraction studies showed that the response of tracheal muscle strips to methacholine was markedly greater than the response of peripheral lung strips, a finding consistent with the QNB binding result. The muscle but not the peripheral lung strip exhibited a relaxing response to epinephrine. Our data indicate a striking quantitative difference in muscarinic and beta adrenergic receptors between lung tissue and tracheal muscle, and that each receptor in the lung is qualitatively similar to the corresponding receptor in the muscle. PMID- 6287141 TI - Effect of alpha-MSH on the corticosteroid production of isolated zona glomerulosa and zona fasciculata cells. PMID- 6287139 TI - Assessment of alpha 2 adrenergic autoreceptor function in humans: effects of oral yohimbine. AB - Administration of three oral doses of yohimbine (10 mg, 15 mg, 20 mg) to eight healthy subjects resulted in significant increases in plasma free 3-methoxy-4 hydroxyphenylethyleneglycol (MHPG). The 15 mg and 20 mg yohimbine doses induced modest increases in systolic blood pressure and autonomic symptoms such as piloerection and rhinorrhea. Marked behavioral effects such as anxiety were not observed. These results indicate that determination of the plasma MHPG response to yohimbine may be of value in assessing alpha 2 adrenergic autoreceptor function in humans. PMID- 6287142 TI - Differentiation between bronchodilation and universal adenosine antagonism among xanthine derivatives. AB - Relaxant effects and adenosine-antagonism of 3-propyl-xanthine (enprofylline) and 10 different methyl-xanthines were examined in isolated guinea-pig tracheas. The chemical structural requirements for tracheal relaxation were found to be different from those for adenosine antagonism by the xanthine derivatives. All xanthines produced relaxation: Enprofylline was about 5 times more potent than theophylline. However, only xanthines with a methyl in the 1-position consistently antagonized the relaxant effect of adenosine. --Theophylline over a wide range of concentrations (30-900 microM) produced a concentration dependent and surmountable antagonism at nervous adenosine receptors (isolated guinea-pig myenteric-plexus preparations). The same concentrations of enprofylline were almost devoid of antagonism at these adenosine receptors. In mice theophylline (6 24 mg/kg given intraperitoneally) dose-dependently increased locomotor activity while enprofylline (2-48 mg/Kg) was without effect on behaviour. "Non-blocking" xanthines such as enprofylline may be potent bronchodilators but lack many theophylline-like actions. We, therefore, forward the hypothesis that universal adenosine antagonism is both unnecessary and undesirable with xanthine antiasthmatics. PMID- 6287143 TI - Opposite temporal changes after a single central administration of B-endorphin: tolerance and sensitization. AB - Rats were given a single unilateral microinjection of B-endorphin in the periaqueductal gray, followed by a second microinjection of the same dose of B endorphin in the same site a week later. A decrease in the analgesic action (i.e., tolerance) but an increase in the hyperthermic action (i.e., sensitization) was observed over this interval. These results suggest that different receptors may mediate these actions of B-endorphin. In addition, these results indicate the need for caution in repeated-measurements studies using this opiate peptide, since the assumption that such temporal effects dissipate within 3-5 days, with resulting minimal carry-over effects from the preceding treatment appears to be unjustified. PMID- 6287144 TI - Forskolin potentiates adrenocorticotropin-induced cyclic AMP production and steroidogenesis in isolated rat adrenal cells. AB - Forskolin, a unique diterpene which directly activates the adenylate cyclase, stimulated production of both cyclic AMP and corticosterone in isolated rat adrenal cells, in vitro. This agent also potentiated the action of adrenocorticotropin and/or cholera toxin on cyclic AMP production and steroidogenesis at lower concentrations. It augmented both an early (cyclic AMP production) and a late (steroidogenesis) action of the hormone in the adrenal gland. PMID- 6287145 TI - A novel pyrazoloquinoline that interacts with brain benzodiazepine receptors: characterization of some in vitro and in vivo properties of CGS 9896. AB - The novel pyrazoloquinoline, CGS, 9896, was a potent inhibitor of specific [3H] flunitrazepam binding in several brain regions with subnanomolar KI values. The inhibition of [3H] propyl beta-carboline-3-carboxylate ([3H]-PCC-) binding by CGS 9896 was enhanced by gamma-aminobutyric acid (GABA) but not by chloride ion. GABA enhancement of CGS 9896 inhibition of [3H]-PCC binding predicts this compound has benzodiazepine (BZD) agonist-type activity. Behavioral studies support this prediction. CGS 9896 was found to protect mice against bicuculline and metrazol induced seizure at doses that did not induce ataxia or sedation. CGS 9896 may represent a class of compounds with potential therapeutic value. The high affinity of this non-BZD compound suggests that CGS 9896 may also be of value as a high affinity ligand for the continued study of BZD receptors. PMID- 6287148 TI - In vivo autoregulation of rat adenohypophyseal thyrotropin-releasing hormone receptor. AB - The possible mechanism of attenuation of thyrotropin response to exogenous thyrotropin-releasing hormone in vivo after repeated administrations of the releasing hormone has been studied. To this end, the effect of prolonged hormone treatment on the binding of hormone to its receptor in the anterior pituitary gland has been evaluated. The data show that prolonged hormonal treatment resulted in a reduction in the number (B max) but no the binding affinity (KD) of the receptor. The effect was reversible and depended on the duration of treatment. This phenomenon of down regulation or the decrease in the receptor number was found not to be due to either the metabolism of releasing hormone or its ability to activate pituitary-thyroid-axis. PMID- 6287146 TI - Effects of extracts of rat brain on the digitalis receptor. AB - There is a possibility that an endogenous substance exists which interacts with a ouabain binding site on Na+, K+-ATPase. Recently, several reports have appeared suggesting the presence of an endogenous digitalis-like substance in acid-acetone extracts of brain. We have demonstrated that in preparing an acid-acetone extract, peroxidized lipids and lysophospholipids are produced, both of which inhibit Na+, K+-ATPase, thereby complicating interpretation. Preliminary evidence suggests, however, that when rat brains are extracted with an aqueous-acetone mixture under nitrogen, a principle is obtained which specifically inhibits Na+, K+-ATPase. PMID- 6287147 TI - The failure of aminophylline to modulate glucagon release in man. AB - There are conflicting results regarding the impact of cyclic AMP on pancreatic glucagon release. The effect of aminophylline, a phosphodiesterase inhibitor, on glucagon secretion was studied in four non-obese, non-diabetic, healthy young male volunteers. The subjects received separate infusions of: 1) aminophylline; 2) aminophylline and propranolol; 3) arginine; 4) aminophylline and arginine; 5) insulin; 6) aminophylline and insulin; and 7) aminophylline and isoproterenol. Aminophylline not only failed to alter glucagon levels but also did not affect the glucagon responses observed after arginine and insulin-induced hypoglycemia. The concurrent infusion of isoproterenol and aminophylline also failed to cause a glucagon response. Although glucagon release has been evoked by cyclic AMP in some in vitro system, administration of aminophylline to human subjects does not enhance secretion. These results indirectly suggest that cyclic AMP is of little importance in the control of glucagon secretion in man, though the effects of aminophylline at the cellular level may be complex. PMID- 6287150 TI - The effect of GABA on in vitro binding of two novel non-benzodiazepines, PK 8165 and CGS 8216, to benzodiazepine receptors in the rat brain. AB - The effect of gamma-aminobutyric acid (GABA) on the binding of PK 8165, a quinoline derivative, and CGS 8216, a pyrazoloquinoline, was assessed in two different regions of the rat brain. PK 8165, a compound with reported anxiolytic properties, inhibited [3H]-propyl beta-carboline-3-carboxylate labeled receptors in the cerebellum with an IC50 of 844 nM and 370 nM in the absence and presence of micro M GABA, respectively. GABA (100 micro M) was less effective in the cerebral cortex, decreasing the IC50 value from 280 to 197 nM. In saturation isotherm studies with [3H]-CGS 8216, a benzodiazepine receptor antagonist, GABA (100 micro M) induced a small but significant reduction in the apparent affinity of [3H]-CGS 8216 for benzodiazepine receptors in the cerebral cortex but the Bmax was unchanged. PMID- 6287149 TI - Pituitary desensitization and the regulation of pituitary gonadotropin-releasing hormone (GnRH) receptors following chronic administration of a superactive GnRH analog and testosterone. AB - We recently demonstrated that chronic daily administration of a superactive GnRH analog to intact rats resulted in an initial stimulation of serum LH levels with a subsequent return of LH levels to baseline at a time when testosterone levels were markedly decreased. These data demonstrated pituitary desensitization following chronic GnRH analog treatment. Administration of GnRH analog with a dose of testosterone which did not markedly lower serum LH levels when administered alone prevented the stimulation of LH secretion by analog. The present studies were undertaken to determine the effects of GnRH analog and testosterone administration on the regulation of pituitary GnRH receptors. Pituitary GnRH receptor binding was increased by analog treatment alone at 20 days and returned to control levels at 40 and 60 days of treatment in parallel to the observed changes in serum LH, demonstrating that one mechanism by which chronic GnRH analog treatment leads to pituitary desensitization is down regulation of pituitary GnRH receptors. Testosterone administration alone decreased pituitary GnRH receptor binding. Combined GnRH analog and testosterone administration prevented the increase in pituitary GnRH receptors observed with analog administration alone. These studies demonstrate that change in pituitary GnRH receptor binding correlate with changes in serum LH and that the stimulatory effects of analog administration on LH are sensitive to inhibition by small doses of testosterone. PMID- 6287151 TI - Comparison of mammary tumorigenesis between litters in relation to mammary tumor virus antigenic expression in the milk of C3H/He mice. AB - As a possible step to estimate the relationship between mammary tumor virus (MTV) and mammary tumorigenesis in mice, MTV antigenic expression in mother's milk and spontaneous mammary tumorigenesis in their daughters were compared between the 1st, the 2nd and the 3rd litters of the highly inbred strains of C3H/H3 mice with low mammary tumor incidence. While MTV antigenic expression was detected in all undiluted samples at each litter by immunodiffusion test, the amount of antigen as measured by the single radial immunodiffusion method was increased with increasing litter numbers. On the other hand, the development of preneoplastic mammary hyperplastic alveolar nodules was different little between litters and mammary tumor incidence by 13 months of age was very low with no difference in all litters. The pattern of estrous cycles and plasma prolactin level were also similar in each litter. The results suggest that spontaneous mammary tumorigenesis in mice is not always affected quantitatively by the amount of MTV when endocrine and genetical conditions are similar. PMID- 6287152 TI - Desensitization of the alpha adrenergic receptor system in guinea pig vas deferens. AB - Desensitization induced by alpha adrenergic (alpha-Ad) stimulation was investigated in organ cultured vas deferens of guinea pig. Brief exposure (1-2 min) of the muscle to noradrenaline (NA) caused short-term desensitization to both NA and acetylcholine (ACh), but not to high K+. After removing the agonist this desensitization completely disappeared within 15 min. Prolonged exposure to NA (i.e., cultured with NA for 3-24 hr) elicited long-term desensitization to NA, ACh and K+ (50 mM), but it did not change the maximal contraction by high K+ (154 mM). After removing NA from the culture medium the response to the agonist was restored to normal within 24 hr, but not within 15 min. The number and affinity of alpha-Ad and muscarinic ACh receptors, which were measured by the binding of 3H-WB4101 and 3H-QNB, respectively, were not changed in the muscle during these treatments. Moreover, long-term desensitization, but not short-term desensitization, was depressed by the concomitant presence of cycloheximide. The possible mechanisms of desensitization were discussed in comparison with those of various receptor systems. PMID- 6287153 TI - [Nephroscintigraphy with 99mTc-labeled pharmaceuticals]. PMID- 6287154 TI - [New diamond bits for orthodontics]. PMID- 6287155 TI - [Testing method for diamond bits for orthodontics]. PMID- 6287156 TI - [Importance, distribution and main causes of accidents in 1980 in two rural health centers in Ivory Coast. Comparative study (author's transl)]. AB - Everywhere, accidents have got some noteworthy consequential effects on socio economical growth. In many developing countries, with agricultural economic background and still limited in their preventive and educational health action, the damage is able to impair their development, as much as malnutrition, infections and parasitic diseases. Carried out according to a precise methodology within a programme of research entitled: "Recueil de statistiques sanitaires et choix d'indicateurs de sante en Cote-d'Ivoire", this present study shows that most of usual accidents to children are caused by active or passive shocks, and adults injuries are due to handling sharp and penetrating tools. Burns and injuries caused by animals are noticeably numerous. Concerning road accidents, their incidence rate increases rapidly, even in traditional rural areas. The high frequency of such accidents burden seriously the weak credits budgeted to health formations of rural or sub-urban districts in developing countries. This only observation should incite leaders in putting in this nosologic entity among the significant sanitary priorities of their respective countries. This study enlightens the fact that the main problems of public health must benefit from a well defined epidemiological approach, as accidents do, in order to determine the priority rank and to justify the action to be taken. So, the planners would be able to explore ways and means which could solve at minimum cost the given problems. Primary health cases seem to be particularly adapted to this. PMID- 6287157 TI - [Abdomen perforating wounds. Study of 61 cases (author's transl)]. AB - A recent increase in the frequency of abdomen perforating wounds has been observed in Ivory Coast, related wih social condition changes. The authors study 61 cases, recorded in 10 years. Most of the cases (62,3 p. 100) are caused by side-arms and 14,8 p. 100 by fire-arms. If 59 p. 100 of the cases resort to criminal assault, 5 p. 100 were caused by suicide attempt; the other cases were consequence of various accidents. The surgical procedures are described. Death rate, with 4 cases, was only of 6,5 p. 100. PMID- 6287158 TI - [Local complications of snake bites (author's transl)]. AB - Morbidity, mortality and local complications of snake bites in the world are discussed. Necrosis follows about 10 p. 100 of snake bites all over the world with important variability from place to place. Aetiologies of necrosis are described. Pharmacology of venoms, local cures and late infections explain diversity of clinical symptomatology of local reactions few of which being severe. Surgical treatment must be applied as soon as possible in a preventive consideration. Local cures such as tourniquet, incision, excision, cauterization, etc., should be abandoned. Local disinfection must be particularly careful treatment of edema or swelling, and antivenom by intravenous route are rather preferable to early surgery. Of course, medical treatment of systemic poisoning will be applied in the same time. PMID- 6287159 TI - [A particular nervous traumatology: lesions of sciatic nerve by quinine intragluteal injections (author's transl)]. AB - These lesions of the sciatic nerve have a significant frequency in Africa, as a consequence of quinine intragluteal injections made by insufficiency trained nurses. Infants and children are most frequently affected. The lesions are a sclerous scar with or without a damage of the neural trunk. The clinical aspect is that of an algo-paralysis. Medical treatment by vitamins, alphachymotrypsine and physiotherapy must be applied early and during two months to have a chance bringing recovery. If it fails, surgical neurolysis is then necessary. Results vary according to the extent of the lesions. PMID- 6287160 TI - [Entomologic and epidemiologic survey on a fatal case of yellow fever occurred in Anyama district (Republic of Ivory Coast)]. PMID- 6287161 TI - [Computer tomography in the diagnosis of the retroperitoneal lymph system]. PMID- 6287162 TI - Fever of undetermined origin: diagnosis and follow-up of 105 cases, 1970-1980. PMID- 6287163 TI - Macrophages: their use in elucidation of the cytoskeletal roles of actin. PMID- 6287165 TI - Reiterated DNA sequences in a mutant strain of Streptomyces glaucescens and cloning of the sequence in Escherichia coli. AB - Streptomyces glaucescens exhibits a high degree of genetic instability. A sequence of 7.2 kb has been found which is present in a few tandemly repeated copies in the wild type strain GLA 0 and is amplified to ca. 500 copies per genome in the mutant strain GLA 1204. This sequence was cloned in Escherichia coli using pBR325 as vector. PMID- 6287164 TI - Genetic map of the crown gall suppressive IncW plasmid pSa. AB - A genetic map of the W incompatibility group plasmid pSa has been prepared through the construction of deletion derivatives of pSa and the cloning of various fragments of pSa in pBR322. Phenotypic analysis of these derivatives has identified the location of genes encoding resistance to chloramphenicol, sulfonamides, spectinomycin, streptomycin, kanamycin, gentamycin, and tobramycin. Information sufficient for the replication of the plasmid in both Escherichia coli and Agrobacterium tumefaciens is contained within a 4 kilobase pair region. Two regions have been identified as involved in the transfer of the plasmid; one of these regions is also involved in the inhibition of oncogenesis by pSa when it is present in an oncogenic strain of A. tumefaciens. Certain of the deletion derivatives of pSa are potential vectors for the cloning and analysis of A. tumefaciens Ti plasmid DNA. PMID- 6287166 TI - R46-derived recombinant plasmids affecting DNA repair and mutation in E. coli. AB - Recombinant plasmids which render their host less mutable and more sensitive to some DNA-damaging agents have been isolated from the N-group plasmid R46. These plasmids have been physically mapped and found to originate from the region of R46 that has been deleted in pKM101. This deleted region is well removed from the muc region of R46 and pKM101 which is responsible for the mutator effects of these plasmids. The effect of these anti-mutagenic plasmids on the ability of pKM101 to complement umuC mutations has been examined, and they have been found to inhibit the complementation of such strains. We propose that these plasmids may code for a negative control function acting on the muc gene. PMID- 6287167 TI - Cloning and expression in Escherichia coli K-12 of the biosynthetic dehydroquinase function of the arom cluster gene from the eucaryote, Aspergillus nidulans. AB - A 1.35 Md DNA HindIII fragment containing part of the arom gene cluster or cluster gene of Aspergillus nidulans encoding biosynthetic dehydroquinase (5 dehydroquinate hydrolyase) has been cloned in plasmid pBR322 on the basis of functional expression in Escherichia coli. The fungal fragment on pBR322, designated pHK29, complements a corresponding E. coli dehydroquinase structural gene (aroD) mutation. pHK29 contains one BamHI, HpaII, PstI, SmaI, XhoI and surprisingly, one HindIII site since pHK29 hybrid Aspergillus DNA is a HindIII fragment itself. The biosynthetic dehydroquinase activity extracted from E. coli strains, containing pHK29, had properties similar to those of the enzyme activity from Aspergillus. The protein specified by pHK29 appears to be 80 kd. No increase of dehydroquinase activity was found in polynucleotide phosphorylase deficient strains (pnp) of E. coli. PMID- 6287168 TI - Cytoplasmic hybridization in Nicotiana: mitochondrial DNA analysis in progenies resulting from fusion between protoplasts having different organelle constitutions. AB - Our previous studies indicated that fusion products with one functional nucleus but organelles of the two fusion partners (i.e. heteroplastomic cybrids) could be obtained by fusing X-irradiated (cytoplasmic donor) with non-irradiated (recipient) Nicotiana protoplasts. The present report deals with the analysis of mitochondria in cybrid populations resulting from the fusion of donor Nicotiana tabacum protoplasts with recipient protoplasts having a N. Sylvestris nucleus but chloroplasts of an alien Nicotiana species, and exhibiting cytoplasmic male sterility. The two fusion parents showed significant differences in restriction patterns of their chloroplast and mitochondrial DNA. Four groups of cybrid plants were obtained by this fusion. All had N. sylvestris nuclei but contained either donor or recipient chloroplasts and had either sterile or fertile anthers. There was no correlation between anther fertility and chloroplast type. The mitochondrial DNA restriction patterns of sterile cybrids were similar to the respective patterns of the sterile fusion partner while the mitochondrial DNA restriction patterns of the fertile cybrids were similar to the respective patterns of the fertile fusion partner. The results indicate an independent assortment of chloroplasts and mitochondria from the heteroplastomic fusion products. PMID- 6287170 TI - Rescue of SV40 following transfection of TC7 cells with cellular DNAs containing complete and partial SV40 genomes. AB - Infectious SV40 virions could be rescued from permissive TC7 cells within one to three subcultures following cotransfection with two cellular DNAs, each containing a complementary portion of the SV40 genome. SV40 virions could also be rescued by transfection of TC7 cells with cellular DNAs from a variety of SV40 transformed cells containing complete genome equivalents but not from cells containing subgenomes alone or defective genomes. Infectious virus was not rescued if the transfecting DNA species was treated with DNAase or if the DEAE dextran pretreatment of the recipient cells was omitted. PMID- 6287169 TI - Regulation of DNA synthesis and capacity for initiation in DNA temperature mutants of Escherichia coli. III. Synthesis of the dnaA protein and of DNA binding proteins. AB - The synthesis and action of the dnaA product with respect to DNA initiation and the synthesis of DNA-binding proteins in Escherichia coli was examined. Results indicate that when dnaA product is irreversibly denatured and must be synthesized before initiation can occur, its synthesis and action appear to be complete approximately 30 min before initiation takes place. However, in mutants whose dnaA product is temperature reversible the action of the dnaA product appears to occur near the time of initiation. Examination of the DNA-binding proteins from the mutants suggests that a 53 kd protein, possibly the dnaA product, may be synthesized at the time of initiation under normal conditions at permissive temperature. The presence of active dnaA product appears to trigger the synthesis of a 60-65 kd protein which may be responsible for preventing another immediate initiation event. PMID- 6287172 TI - Continuous synthesis of the dnaA gene product of Escherichia coli in the cell cycle. AB - The dnaA gene product of Escherichia coli, identified as a weakly basic protein of about 48,000 daltons (Yuasa and Sakakibara 1980), can be separated from other cellular proteins by means of two-dimensional gel electrophoresis. Synthesis of the dnaA protein took place continuously during a cell growth cycle. The newly synthesized dnaA protein persisted stably for one generation. Thermosensitive dnaA protein produced by the dnaA167 mutant was stable at 30 degrees C, but was disintegrated at 42 degrees C. The amount of intact dnaA protein present in the mutant exposed to the high temperature for 60 min was less than a quarter of the amount at the time of the shift. The cells having the reduced amount of intact dnaA protein were capable of initiating a new round of chromosome replication at the low temperature without de novo synthesis of the dnaA protein. The potential of the mutant for initiation of DNA replication decreased with reduction in the amount of the thermoreversible dnaA protein. The mutations dnaA167 and dnaA46 had no significant effect on the syntheses of the dnaA mRNA and the protein product at the low and high temperatures. PMID- 6287171 TI - Discontinuity of homology of Escherichia coli and Salmonella typhimurium DNA in the lac region. AB - Partial homology of Salmonella typhimurium DNA to Escherichia coli DNA was demonstrated by Southern hybridization blots to exist on either side of the lac operon of E. coli but no homology was detected between S. typhimurium DNA and about 12 kb of E. coli DNA including the lac genes as well as about 5 kb of E. coli DNA between lac and proC. Thus portions of DNA seem to have been either added to the E.coli genome or deleted from the S. typhimurium genome since their divergence from a common ancestor. Although an IS1 element was located near the lac operon of E. coli, the insertional element was shown not to be near any of the junctures of discontinuity of E. coli--S. typhimurium homology near lac. PMID- 6287173 TI - Apurinic endonuclease from Drosophila melanogaster: reduced enzymatic activity in excision-deficient mutants of the mei-9 and mus(2)201 Loci. AB - An endonuclease activity, which is specific for partially depurinated PM2 DNA, has been assayed in extracts prepared from cultured cells and larval brain ganglia of Drosophila melanogaster. Activity detected in repair-proficient cells is stimulated by Mg++ and is inhibited by EDTA. Extracts prepared from established cell cultures of the excision-deficient strain mei-9a and from larval brain ganglia derived from y mei-9a, mei-9L1, sn3 mei-9D2 and sn3 mei-9D4 are partially deficient in this activity. At least one allele of a second excision deficient strain, cn mus (2)201D1, also shows reduced AP-endonuclease activity in ganglia extracts. PMID- 6287175 TI - Adenosine triphosphatase activity of Ureaplasma urealyticum. AB - Adenosine triphosphatase activity of U. urealyticum is an integral membrane-bound protein which cannot be detached from the membrane by mild treatment with EDTA in low-ionic strength media nor by ionic detergents which rapidly inactivated the enzyme. The enzyme was Mg++ dependent; Mn++ and Co++ could replace Mg++ to some extent. A slight stimulatory effect was also exerted by sodium and lithium. The enzyme showed a nucleotide triphosphatase activity, but ADP was hydrolyzed at close to 40% the rate of ATP and other nucleotide monophosphatase were hydrolyzed at a very slow rate. Oubain and oligomycin did not inhibit the adenosine triphosphatase activity, whereas DCCD, NBD-Cl and several sulfhydryl-blocking reagents strongly reduced its activity. The enzyme could not be stimulated by trypsin pretreatment. It seems that the complex enzyme is tightly linked to the lipid bilayer of the membrane and differs in many aspects from the F0-F1 (Mg++, Ca++)-ATPase of bacteria. PMID- 6287174 TI - The regulation of the ammonia assimilatory enzymes in Rel+ and Rel- strains of Salmonella typhimurium. AB - The influence of the relA1 mutation on the regulation of the ammonia assimilatory enzymes, glutamate dehydrogenase (EC 1.4.1.4), glutamine synthetase (EC 6.3.1.2), and glutamate synthase (EC 1.4.1.3), was examined. When cells grown in rich media (either Luria broth or glucose-ammonia plus casamino acids) were transferred to a glucose-ammonia medium, the relA mutant failed to resume growth and did not have the same increase in any of the assimilatory enzyme activities as the rel+ strain. This effect was particularly dramatic for glutamate dehydrogenase, which increased 6-fold in the rel+ strain. Measurements of the guanosine nucleotide concentrations showed that the rel+ strain had a ppGpp concentration about 9 times that of the relA mutant 5 min after the shift to minimal medium. These results are consistent with those for other biosynthetic enzymes and show that the ammonia assimilatory enzymes require a relA product for their synthesis during shift from rich to minimal media. In addition, we examined the response of these strains to a change in nitrogen source. The relA mutant again failed to resume growth after a shift from glucose-ammonia to glucose-arginine medium. Even though the ppGpp concentration did not increase, the rel+ strain grew and increased glutamine synthetase activities about 2-fold. These changes the absence of increased ppGpp levels suggest that some other relA-mediated function is important during this change in nitrogen source. PMID- 6287176 TI - Human cytomegalovirus cycle in synchronized cells: electron microscope study. AB - Human Cytomegalovirus (HCMV) was inoculated in synchronized human fibroblasts at different phases of the cell cycle. the virus replication appeared strongly dependent on the host metabolic state, being faster when the infection was carried out in G2 and S phase. PMID- 6287177 TI - Repression of adenylate cyclase in the genus Bordetella. AB - Virulent phase I strains of Bordetella pertussis synthesize the regulatory metabolite adenosine 3',5'-cyclic monophosphate (cAMP); whereas, non-virulent phase IV strains are unable to do so. These findings were confirmed and extended using additional strains of B. pertussis as well as strains of Bordetella parapertussis and Bordetella bronchiseptica. Levels of adenylate cyclase activity and subsequent cAMP production were found to correlate. High adenylate cyclase activity during exponential phase was followed by an accumulation of cAMP during late log and throughout stationary phase. Measurements obtained during the X- to C-mode transition process indicated that adenylate cyclase is subject to complete repression. PMID- 6287178 TI - Antimicrobial effect of human serum IgA. AB - Serum IgA, IgG and colostrum secretory IgA prepared from specimens pooled from a large number of human beings were shown to have measurable levels of antibodies against Escherichia coli, Pseudomonas aeruginosa, Klebsiella Pneumoniae, poliovirus, Coxsackie B virus, echovirus and influenza virus. Serum IgA exerted a bacteriostatic effect in vitro on E. coli and P. aeruginosa, which increased in the presence of the iron-binding proteins lactoferrin and transferrin. This bacteriostasis was reduced when the iron-binding proteins were saturated with iron. Similar results were obtained with IgG and secretory IgA. The bacteriostatic effect of serum IgA was also shown in vivo, in the peritoneal cavity of mice. The effect was suppressed by iron. Iron-chelating substances, siderophores, excreted by E. coli diminished the co-operative bacteriostatic effect of serum IgA and transferrin. Siderophore production by E. coli was inhibited in the presence of serum IgA, but not when serum IgA was deprived of specific antibody by absorption with E. coli. These results indicate that serum IgA has a potent bacteriostatic effect in co-operation with transferrin or lactoferrin because of the inhibitory effect of the specific antibody on siderophore production by E. coli. PMID- 6287179 TI - Two distinct electrophoretic migration patterns of RNA segments of human rotaviruses prevalent in Japan in relation to their serotypes. PMID- 6287180 TI - Assembly of paramyxoviruses. PMID- 6287181 TI - Purification of Rickettsia tsutsugamushi by Percoll density gradient centrifugation. AB - Purification of Rickettsia tsutsugamushi has been achieved by Percoll density gradient centrifugation. The microorganisms purified showed good retention of infectivity and intracellular morphology. Budding rickettsiae in the egressing stage and intracellular rickettsiae in the multiplying process were harvested separately and purified by this technique. In electron microscopic observations, the intracellular rickettsiae obtained were surrounded with double membrane layers of cell wall and cell membrane, and the budding rickettsiae were enveloped with an additional outermost membrane which may have originated from host cell membrane obtained in the budding process. PMID- 6287183 TI - Universal adenosine receptor antagonism is neither necessary nor desirable with xanthine antiasthmatics. AB - The great diversity of pharmacological effects of xanthines may well reflect different cellular mechanisms of action. Major attention is presently devoted to adenosine receptor antagonism that, in contrast to phosphodiesterase inhibition, is clearly produced by therapeutic concentrations of theophylline. The ubiquitous adenosine effects together with the universal and potent blocking action of methylxanthines have led investigators to believe that most pharmacological actions including antiasthmatic effects of theophylline reflect adenosine antagonism. The present hypothesis proposes that universal adenosine receptor antagonism is neither necessary nor desirable with xanthine antiasthmatics. Supporting the hypothesis a xanthine derivative that seems to be devoid of functional effects at important adenosine receptor sites has been shown to be a potent bronchodilator drug that lacks theophylline-like diuretic and CNS stimulant behavioural effects. PMID- 6287182 TI - Role of Lyt-1 positive immune T cells in recovery from herpes simplex virus infection in mice. PMID- 6287184 TI - Some theoretical questions of the peptide and steroid hormone regulation. Part III.: The RLVC hypothesis as a possible mode for the interaction of the peptide and steroid hormone regulation. AB - My previous models for peptide and steroid hormone actions, in accordance with their biological and molecular-informational requirements, suggested possible mechanisms for the subcellular interactions of the two regulatory systems. In my hypothesis the homogeneous or heterogeneous multiligand binding-proteins (receptors, characterized previously) have the ability to bind both steroids and (cyclic) nucleotide phosphates in a unique distribution and sequence. The specificity of such a Receptor-Ligand Variable Complex (RLVC), the pattern of the steroid and/or (cyclic) nucleotide phosphate ligands, is determined by the equilibrium between the actually acting steroids and peptides on the one hand and by the binding possibilities of the receptor on the other. PMID- 6287185 TI - The pathogenesis of systemic lupus erythematosus. AB - Many investigators believe that systemic lupus erythematosus is an autoimmune disease, perhaps caused by inadequate suppressor T lymphocyte activity, which permits the activation of autoantibody producing B lymphocytes. This paper discusses the testable hypothesis that a superoxide-generating, chromosome aberration-inducing factor (clastogenic factor), present in the lymphocytes of lupus patients but absent from normals, is responsible for such a suppressor cell defect. Superoxide or activated oxygen species derived from it, such as hydroxyl radical, may be the molecular mediators of CF activity. PMID- 6287186 TI - Acute haemorrhagic cystitis caused by adenovirus type 11 in a recipient of a transplanted kidney. AB - A case of haemorrhagic cystitis caused by adenovirus type 11, which occurred in a female patient 13 weeks after renal transplantation is described. The patient lacked neutralising antibody to adenovirus type 11 before transplantation, but this antibody was present in the serum of the kidney donor. The patient developed antibody to adenovirus type 11, and this virus was isolated from her throat and urine. The data suggest that the infection was transmitted by the kidney graft. PMID- 6287187 TI - [Neurogenic changes in the end interval of the ECG]. PMID- 6287188 TI - Lung cancer as a complication of prolonged survival in patients with lymphoma. AB - As a result of successful therapy with radiation, chemotherapy, or both, patients with lymphoma are living longer. The improved survival has led to an increased awareness of long term complications including second or subsequent malignancies. We have recently seen six cases of lung cancer--three adenocarcinomas, one squamous cell carcinoma, and two small cell carcinomas--among 655 patients with Hodgkin and non-Hodgkin lymphoma suggesting a possible association, although other carcinogenic or cocarcinogenic factors cannot be eliminated. PMID- 6287189 TI - Late effects of treatment of cancer in infancy. AB - Eighty-six children were diagnosed with cancer in infancy, followed for at lest 5 years, and assessed for late effects of disease and therapy. One child subsequently died from respiratory failure and 3 died from second primary cancers. Another patient survived second primary cancers of the skin. The high frequency of new cancers (4 observed, 0.09 expected) was attributable to host susceptibility factors and treatment effects. Kyphoscoliosis was diagnosed in 44 patients, 40 of whom had received radiotherapy to the spine. Other patients had neurologic deficits, pulmonary fibrosis, hypoplastic breasts, bowel adhesions, thyroid nodules, musculoskeletal defects, and liver fibrosis associated with tumor therapy. Sequelae of cancer were more common after treatment in infancy than in later childhood. Improved treatments and knowledge of natural history can reduce adverse effects of therapy. PMID- 6287192 TI - Population-based studies of diabetes mellitus in Minnesota: rationale and methods. PMID- 6287190 TI - Treatment of small cell lung cancer: intensive remission induction, radiation therapy, and maintenance chemotherapy. AB - Forty-eight patients with small cell lung cancer were treated with an intensive therapy regimen which included induction chemotherapy with vincristine, adriamycin, procarbazine, and methotrexate. This was followed by radiation therapy given to areas of residual disease after restaging. Maintenance chemotherapy with cyclophosphamide, methotrexate, and procarbazine was then given to patients who had entered complete remission (CR). Fifteen out of 17 patients with limited disease (88%) achieved complete remission. Remission rates for patients with extrathoracic metastases were somewhat lower. Seven of 11 patients (64%) of patients with a single extrathoracic metastatic site achieved CR, while only 6/20 (30%) with multiple extrathoracic metastases went into remission. Median survival for patients who achieved complete remission was 17 months. PMID- 6287193 TI - Allosteric coupling between morphine and enkephalin receptors in vitro. AB - In a recent paper from our laboratory [Mol. Pharmacol 21:538-547 (1982)] evidence was presented which suggested that [3H]leucine enkephalin labels a single class of binding sites (the enkephalin receptor) and that morphine allosterically induces a masking of enkephalin receptors as a consequence of binding to a receptor (the morphine receptor) not labeled by the 3H-peptide. Evidence is presented in this paper that [3H]etorphine can be used to label selectively the morphine receptor and that the inhibitory dissociation constants (KI) of morphine, etorphine, and human beta-endorphin for the [3H]etorphine binding site closely approximate the concentration of these drugs which produce a half-maximal decrease in the number of enkephalin receptors. Furthermore, an examination of the interaction of leucine enkephalin and methionine enkephalin with the morphine receptor has demonstrated that the pentapeptides are not competitive inhibitors of [3H] etorphine binding, and that they have much lower affinities for the morphine receptor than previously thought. On the basis of these data, a working hypothesis has been formulated which postulates that distinct morphine and enkephalin receptors coexist in an opioid-receptor complex. PMID- 6287191 TI - Partial hypothalamic insufficiency resulting from herpes simplex encephalitis: report of a probable case. PMID- 6287194 TI - Dimeric enkephalins display enhanced affinity and selectivity for the delta opiate receptor. PMID- 6287195 TI - The opiate receptor-binding interactions of opiate alkaloids and of an opioid peptide in rat brain membranes. Selection by manganese ions and by cholic acid (sodium salt) and minimalization of cross-reaction in vitro. PMID- 6287196 TI - Differential modification of the interaction of cardiac muscarinic cholinergic and beta-adrenergic receptors with a guanine nucleotide binding component(s). PMID- 6287197 TI - Sodium inhibits both adenylate cyclase and high-affinity 3H-labeled p aminoclonidine binding to alpha 2-adrenergic receptors in purified human platelet membranes. PMID- 6287198 TI - Heterogeneity of brain benzodiazepine receptors demonstrated by [3H]propyl beta carboline-3-carboxylate binding. PMID- 6287199 TI - Structural and conformational properties of peptides interacting with the glutathione receptor of hydra. AB - We have made a detailed investigation of structure-activity relationships of the glutathione-induced feeding response in the fresh water coelenterate, Hydra attenuata, to map structural and conformational properties of feeding response agonists. We find that the gamma-glutamyl residue of glutathione contains essential binding sites for receptor interaction, without which antagonistic as well as agonistic properties are lost. Any structural alteration which perturbs either the alpha-amino or the alpha-carboxyl group or their relative spatial orientations within the peptide has yielded an inactive derivative. An absolute requirement for activation of the receptor is a second-residue side chain of the appropriate size; analogues with second-residue side chains too large or too small are inhibitory. On the basis of the activity of conformational analogues of glutathione, torsional angles for the second residue equal to those of a right handed alpha-helix are compatible with stimulus generation. PMID- 6287200 TI - Site of action of phencyclidine. IV. Interaction of phencyclidine and its analogues on ionic channels of the electrically excitable membrane and nicotinic receptor: implications for behavioral effects. PMID- 6287201 TI - Stimulation of neuroblastoma adenylate cyclase by arachidonic acid metabolites. PMID- 6287202 TI - Inhibition of aggregation and stimulation of cyclic AMP generation in intact human platelets by the diterpene forskolin. PMID- 6287203 TI - Glucagon stimulation of hepatic Na+, K+-ATPase. AB - In the perfused rat liver administration of glucagon was shown to result in a transiently increased uptake of K+, indicating the possible involvement of the Na+, K+-ATPase. Direct measurement of the activity of Na+, K+-ATPase revealed a two-fold stimulation of the enzyme by glucagon. The effect of glucagon on the activity of the enzyme was immediate. Simultaneously with the increase in the activity of the Na+, K+-ATPase, the activity of Mg2+-ATPase decreased. In order to evaluate whether the activation of the Na+, K+-ATPase by glucagon is related to the metabolic effects of the hormone, experimental conditions known to interfere with the activity of the enzyme were employed and glucagon stimulation of Ca2+-efflux, mitochondrial metabolism and gluconeogenesis were measured. K+ free perfusate, high K+ perfusate or ouabain interfered to varying degrees with the glucagon stimulation of these responses. The combination of K+-free perfusate and ouabain almost completely abolished the glucagon stimulation of all three parameters. These results demonstrate the glucagon stimulation of Na+, K+-ATPase and raise the possibility that the activation of the enzyme by glucagon might be a necessary link for the manifestation of its metabolic effects. PMID- 6287205 TI - Enzymatic oxidation of alpha-1-proteinase inhibitor in abnormal tissue turnover. PMID- 6287207 TI - Protein kinase activities in Neurospora crassa. Molecular properties. PMID- 6287206 TI - DNA glycosylases. AB - Various DNA glycosylases exist, which initiate the first step in base-excision repair. A summary of the kinetic and physical characteristics of three classes of DNA glycosylases are presented here. The first class discussed, include glycosylases which recognize alkylated DNA. Various data from enzymes derived from both prokaryotic and eukaryotic sources is discussed. The second class deals with a glycosylase that recognizes and initiates the excision of pyrimidine dimers in DNA. To date, this enzyme has only been uncovered from two sources, Micrococcus luteus and the T4 bacteriophage of E. coli. The third class consists of the most studied of the glycosylases, the uracil-DNA glycosylase enzymes. Various characteristics are presented for the uracil-DNA glycosylases derived from various sources. Recent information from our laboratory is presented implicating that herpes simplex virus may mediate a uracil-DNA glycosylase activity in productivity infected cells. PMID- 6287208 TI - Low Km cyclic AMP phosphodiesterase of yeast may be bound to ribosomes associated with the nucleus. AB - The sub-cellular distribution of low Km cyclic AMP phosphodiesterase (defined as the EDTA-sensitive activity at 1 microM cyclic AMP) was examined using spheroplast lysates and mechanical disintegrates of yeast. Close to 65% of the enzyme was particle-bound in each case. Most of the bound activity in mechanical disintegrates sedimented at 145 000 g in an RNA-rich fraction, and could be solubilised from this fraction by RNase treatment. With spheroplast lysates, however, 50% of the enzyme co-sedimented with DNA at 5 000 g, and the highest specific activity was in purified nuclei with a protein/DNA mass ratio of 16. The results suggest that at least 50% of the enzyme is bound by ribosomes attached to the outer nuclear membrane. PMID- 6287204 TI - Calmodulin. PMID- 6287209 TI - Calcium transport by cardiac sarcoplasmic reticulum and phosphorylation of phospholamban. PMID- 6287211 TI - Plasminogen activator: analysis of enzyme induction by ultraviolet irradiation mapping. AB - Ultraviolet irradiation mapping techniques have previously been used to study the organization of eucaryotic gene classes and transcription units. We used the same method to probe some regulatory phenomena observed in the induction of plasminogen activator (PA) biosynthesis: PA synthesis in chicken embryo fibroblasts is induced by tumor-promoting phorbol esters and by retinoic acid; furthermore, PA induction by phorbol esters is synergistic with transformation, being 10- to 20-fold greater in virus-transformed cells than in normal cells. We found that the ultraviolet irradiation inactivation cross sections for PA induction by phorbol esters and by retinoate differed significantly, suggesting that these agents induce PA biosynthesis by different mechanisms. On the other hand, the ultraviolet irradiation sensitivity of phorbol ester induction in normal chicken embryo fibroblasts was the same as in transformed cells, indicating that the synergism of transformation and phorbol esters is probably not due to different pathways of PA induction. PMID- 6287210 TI - Fine structure and evolution of the rat serum albumin gene. AB - The exons, their boundaries, and approximately half of the intronic deoxyribonucleic acid of the rat serum albumin gene were sequenced. In addition to the 14 exons identified earlier by R-loop analysis, a small exon was detected between the "leader" exon (Z) and exon B. The leader exon encoded the 5' untranslated portion of albumin messenger ribonucleic acid and the "pre-pro" oligopeptide present on the nascent protein. The sites of initiation and termination of transcription were tentatively identified by comparison of the 5' and 3' gene-flanking sequences with those of other eucaryotic genes. All 28 intron/exon junctions conformed to the "GT-AG rule" (Breathnach et al., Proc. Natl. Acad. Sci. 75:4853-4857, 1978). The three homologous domains of albumin were encoded by three subgenes that consisted of four exons each and evolved by intragenic duplication of a common ancestor. The second and forth exons of each subgene appeared to be the result of an even earlier duplication event. We propose a model for the evolution of this gene that accounts for the observed patterns of exon size and homology. PMID- 6287214 TI - Organization and closing of mitochondrial deoxyribonucleic acid from Paramecium tetraaurelia and Paramecium primaurelia. AB - Previously we showed that the mitochondrial deoxyribonucleic acid (DNA) from Paramecium aurelia consists of a linear genome and that replication of this genome is initiated at one terminus and proceeds unidirectionally to the other terminus. Analyses of mitochondria from four closely related species (1, 4, 5, and 7) indicated that the species 1, 5, and 7 DNAs are essentially completely homologous but that the species 4 mitochondrial DNA is only 40 to 50% homologous with that from species 1. The major regions of homology are those containing the genes for ribosomal ribonucleic acid (RNA). To understand the replication and organization of the linear mitochondrial genome better, we compared species 1 (Paramecium primaurelia) and 4 (Paramecium tetraaurelia) DNAs with regard to restriction fragment mapping and homology between initiation regions; we also identified the sites of the genes for ribosomal RNA. In general, the structures of the species 1 and 4 mitochondrial genomes were quite similar. Each ribosomal RNA gene was present in one copy per genome, with the large ribosomal RNA gene located near the terminal region of replication and the small ribosomal RNA gene located more centrally. These two genes were separated by about 10 kilobases in the species 1 genome and by about 12 kilobases in the species 4 genome. In contrast to our previous findings, by using nonstringent hybridization conditions we detected homology between the species 1 and 4 DNA fragments containing the initiation regions. We constructed recombinant DNA clones for many fragments, especially those containing the initiation region and the ribosomal RNA genes. We also constructed restriction enzyme maps for six enzymes for both P. primaurelia and P. tetraaurelia. PMID- 6287212 TI - Sequential transcription-translation of simian virus 40 by using mammalian cell extracts. AB - Ribonucleic acids (RNAs) transcribed in vitro by using the whole-cell extract system of Manley et al. (Proc. Natl. Acad. Sci. U.S.A. 77:3855-3859, 1980) were tested for their efficiency and fidelity in directing protein synthesis in reticulocyte lysates. Simian virus 40 deoxyribonucleic acid (DNA), cleaved by various restriction endonucleases, was used as the template. Successful translation of the small tumor antigen t, as well as the capsid proteins VP1, VP2, and VP3, was detected by immunoprecipitation analysis. Although no synthesis of large T antigen was detected, use of this technology allows detection of large T synthesis resulting from the correct splicing of as little as 0.2% of the in vitro RNA transcripts, making it ideal for use as an in vitro splicing assay. Transcripts synthesized in vitro were used as messages at least as efficiently as were viral messenger RNA's (mRNA's) synthesized in vivo; and in the case of small t, there was more efficient translation of small t mRNA synthesized in vitro than of small t mRNA synthesized in vivo. The transcripts that served as mRNA's for the various polypeptides were identified by using the following two criteria. (i) The sensitivity of synthesis of a given protein to digestion of the template DNA with restriction enzymes allowed the localization of the promoter and coding regions. (ii) Translation of size-fractionated RNA allowed confirmation of the transcript-mRNA assignments. With these techniques we found that VP2, VP3 and, in some cases, VP1 synthesis resulted from the initiation of translation at internal AUG codons. In fact, families of polypeptides were produced by initiation of translation at AUG codons within sequences coding for VP1 and T, presumably as a result of transcription initiation events that generated 5' ends immediately upstream from these AUGs. Application of this technology for the identification of coding regions within cloned DNA fragments is discussed. PMID- 6287213 TI - Comparison between the viral transforming gene (src) of recovered avian sarcoma virus and its cellular homolog. AB - Recovered avian sarcoma viruses are recombinants between transformation-defective mutants of Rous sarcoma virus and the chicken cellular gene homologous to the src gene of Rous sarcoma virus. We have constructed and analyzed molecular clones of viral deoxyribonucleic acid from recovered avian sarcoma virus and its transformation-competent progenitor, the Schmidt-Ruppin A strain of Rous sarcoma virus. A 2.0-megadalton EcoRI fragment containing the entire src gene from each of these clones was subcloned and characterized. These fragments were also used as probes to isolate recombinant phage clones containing the cellular counterpart of the viral src gene, termed cellular src, from a lambda library of chicken deoxyribonucleic acid. The structure of cellular src was analyzed by restriction endonuclease mapping and electron microscopy. Restriction endonuclease mapping revealed extensive similarity between the src regions of Rous sarcoma virus and recovered avian sarcoma virus, but striking differences between the viral src's and cellular src. Electron microscopic analysis of heteroduplexes between recovered virus src and cellular src revealed a 1.8-kilobase region of homology. In the cellular gene, the homologous region was interrupted by seven nonhomologous regions which we interpret to be intervening sequences. We estimate the minimum length of cellular src to be about 7.2 kilobases. These findings have implications concerning the mechanism of formation of recovered virus src and possibly other cell-derived retrovirus transforming genes. PMID- 6287216 TI - Three segments from the monkey genome that hybridize to simian virus 40 have common structural elements. AB - Three cloned segments that hybridize to a region of simian virus 40 (SV40) deoxyribonucleic acid including the origin of replication have been isolated from a monkey genomic library. The primary structure of one segment was previously reported (T. McCutchan and M. Singer, Proc. Natl. Acad. Sci. U.S.A. 78:95-99, 1981). We report here the sequences of the other two segments and a comparison of all three. The SV 40-hybridizing region in each segment is limited to several hundred base pairs. All of the segments contain multiple and disconnected sequences homologous to the region of SV40 directly surrounding the viral replication origin. The number and arrangement of the homologous sequences is different in the three segments. However, the segments have the following features in common: (i) each contains multiple copies of the sequence GGGCGGPuPu, which also appears six times near the origin of SV40; (ii) each contains several strong homologies to the central dyad symmetry of SV40; (iii) each contains a long internal repeat, as does the origin region of SV40. The three SV40 hybridizing segments are members of a larger family of genomic sequences that hybridize well to each other, but not necessarily to SV40. PMID- 6287218 TI - Transposition of yeast mating type genes from two translocations of the left arm of chromosome III. AB - In the yeast Saccharomyces cerevisiae, the HIS4C gene lies on the left arm of chromosome III. We analyzed two chromosomal rearrangements that have HIS4C translocated either to chromosome XII or to a new translocation chromosome. Using the cmt mutation that allows expression of the normally silent copies of mating type genes, we found that both of these translocations also carried HML alpha, more than 30 map units distal to HIS4C which normally lies on chromosome III. In the case of the translocation chromosome (designated T3), we also found an exchange event between HML alpha on the translocation chromosome and HMLa on chromosome III. In diploids containing two T3 chromosomes (one carrying HML alpha and the carrying HMLa), we found that HML was 32 centimorgans from HIS4C, which was 10 centimorgans from an unknown centromere. In homothallic strains carrying HMLa MATa HMRa on chromosome III, switching from MATa to MAT alpha could occur by using the HML alpha on the translocation as the sole donor of alpha information. Transposition from HML alpha on chromosome T3 was about 20 to 40% as efficient as transposition from intact chromosome III. In contrast, transposition from the HML alpha inserted into chromosome XII was reduced about 100-fold. This reduced efficiency did not appear to be caused by an alteration in the sequences immediately surrounding HML alpha in the translocation. The translocated HML alpha sequence was located in the same size (29-kilobase) SalI fragment as was found in chromosome III, and the same EcoRI, HindIII, and BglII restriction sites were also found. Furthermore, HML alpha was still under the control of the CMT gene, which maintains HML as a silent copy of mating type information. These results suggested that the position of the HML alpha sequence plays an important role in the efficiency of mating type switching. PMID- 6287215 TI - Nonlytic simian virus 40-specific 100K phosphoprotein is associated with anchorage-independent growth in simian virus 40-transformed and revertant mouse cell lines. AB - Normal fibroblasts display two distinct growth controls which can be assayed as requirements for serum or for anchorage. Interaction of mouse 3T3 fibroblasts with simian virus 40 (SV40) thus generates four classes of transformed cells. We have examined viral gene expression in these four classes of cell lines. Immunoprecipitation of [35S]methionine-labeled cell extracts with an antiserum obtained from tumor-bearing hamsters detected the SV40 large T and small t proteins (94,000 molecular weight [94K], 17K) and the nonviral host 54K protein in all cell lines tested. A tumor antigen with an apparent molecular weight of 100,000 was also found in some, but not all, lines. Similar "super T" molecules have been found by others in many rodent transformed lines. We carried out an analysis of the relation of phenotype to relative amounts of these proteins in cell lines of the four classes, using the Spearman rank correlation test. The amount of the 100K T antigen relative to the 94K T antigen or to total viral protein was well correlated with the ability to form colonies in semisolid medium. No significant correlation was found between quantities of labeled 94K T antigen, 54K host antigen, or 17K t antigen and either serum or anchorage independence. Mouse cells transformed with the small t SV40 deletion mutant 884 synthesized a 100K T antigen, suggesting that small t is not required for the production of this protein. The 100K T antigen migrated more slowly than lytic T. Since mixtures of extracts from cells expressing and lacking the 100K T antigen yielded the expected amount of this protein, it is unlikely that the 100K T derives from the 94K protein by a posttranslational modification. PMID- 6287217 TI - Relationship of amplified dihydrofolate reductase genes to double minute chromosomes in unstably resistant mouse fibroblast cell lines. AB - Murine 3T6 selected in increasing concentrations of methotrexate were unstable with respect to dihydrofolate reductase overproduction and methotrexate resistance when they are cultured in the absence of methotrexate. An analysis of the karyotypes of these resistant cells revealed the presence of numerous double minute chromosomes. We observed essentially identical kinetics of loss of dihydrofolate reductase gene sequences in total deoxyribonucleic acid and in deoxyribonucleic acid from fractions enriched in double minute chromosomes and in the numbers of double minute chromosomes per cell during reversion to methotrexate sensitivity, and this suggested that unstably amplified gene sequences were localized on double minute chromosomes. This conclusion ws also supported by an analysis of cell populations sorted according to dihydrofolate reductase enzyme contents, in which relative gene amplification and double minute chromosome content were related proportionally. PMID- 6287219 TI - Sequence heterogeneity, multiplicity, and genomic organization of alpha- and beta tubulin genes in sea urchins. AB - We analyzed the multiplicity, heterogeneity, and organization of the genes encoding the alpha and beta tubulins in the sea urchin Lytechinus pictus by using cloned complementary deoxyribonucleic acid (cDNA) and genomic tubulin sequences. cDNA clones were constructed by using immature spermatogenic testis polyadenylic acid-containing ribonucleic acid as a template. alpha- and beta-tubulin clones were identified by hybrid selection and in vitro translation of the corresponding messenger ribonucleic acids, followed by immunoprecipitation and two-dimensional gel electrophoresis of the translation products. The alpha cDNA clone contains a sequence that encodes the 48 C-terminal amino acids of alpha tubulin and 104 base pairs of the 3' nontranslated portion of the messenger ribonucleic acid. The beta cDNA insertion contains the coding sequence for the 100-C terminal amino acids of beta tubulin and 83 pairs of the 3' noncoding sequence. Hybrid selections performed at different criteria demonstrated the presence of several heterogeneous, closely related tubulin messenger ribonucleic acids, suggesting the existence of heterogeneous alpha- and beta-tubulin genes. Hybridization analyses indicated that there are at least 9 to 13 sequences for each of the two tubulin gene families per haploid genome. Hybridization of the cDNA probes to both total genomic DNA and cloned germline DNA fragments gave no evidence for close physical linkage of alpha-tubulin genes with beta-tubulin genes at the DNA level. In contrast, these experiments indicated that some genes within the same family are clustered. PMID- 6287220 TI - Cellular Moloney murine sarcoma (c-mos) sequences are hypermethylated and transcriptionally silent in normal and transformed rodent cells. AB - Moloney murine sarcoma virus carries an oncogenic sequence (v-mos) which is homologous to a single copy gene (c-mos) present in the normal cells of several vertebrate species. Because of the possible significance of c-mos sequences in normal development and malignant transformation induced by physical or chemical agents, we have examined the state of integration, methylation, and transcriptional activity of c-mos sequences in a variety of normal rodent tissues, normal cell lines, or cell lines transformed by radiation or chemical carcinogens. DNA-DNA hybridization, utilizing the Southern blotting technique and a plasmid-derived DNA probe representing the v-mos sequence, gave no evidence for rearrangements of the c-mos sequence in the DNAs obtained from these diverse cell types. Parallel studies employing the restriction enzyme isoschizomers HpaII and MspI indicated that in all of these cell types the c-mos sequences were heavily methylated. In addition, analysis of cellular RNAs by blot hybridization with the v-mos probe failed to detect evidence of transcription of the c-mos sequences in any of these cell types. This was in contrast to a Moloney sarcoma virus transformed cell line in which we found that the integrated v-mos sequence was both undermethylated and extensively transcribed. Thus, it would appear that c mos sequences do not play a role in the transformation of rodent cells by chemical or physical agents, although the possible role of other endogenous onc sequences remains to be determined. PMID- 6287221 TI - Conditions necessary for inhibition of protein synthesis and production of cytopathic effect in Aedes albopictus cells infected with vesicular stomatitis virus. AB - The relationship between the development of cytopathic effect (CPE) and the inhibition of host macromolecular synthesis was examined in a CPE-susceptible cloned line of Aedes albopictus cells after infection with vesicular stomatitis virus. To induce rapid and maximal CPE, two conditions were required: (i) presence of serum in the medium and (ii) incubation at 34 degrees C rather than at 28 degrees C. In the absence of serum, incubation of infected cultures at 34 degrees C resulted in a significant increase in viral protein and RNA synthesis compared with that observed at 28 degrees C. However, when serum was present in the medium, by 6 h after infection protein synthesis (both host and viral) was markedly inhibited when infected cells were maintained at 34 degrees C. RNA synthesis (host and viral) was also inhibited in vesicular stomatitis virus infected cells maintained at 34 degrees C with serum, but somewhat more slowly than protein synthesis. Examination of polysome patterns indicated that when infected cultures were maintained under conditions which predispose to CPE, more than half of the ribosomes existed as monosomes, suggesting that protein synthesis was being inhibited at the level of initiation. In addition, the phosphorylation of one (or two) polysome-associated proteins was reduced when protein synthesis was inhibited. Our findings indicate a strong correlation between virus-induced CPE in the LT-C7 clone of A. albopictus cells and the inhibition of protein synthesis. Although the mechanism of the serum effect is not understood, incubation at 34 degrees C probably predisposes to CPE and inhibition of protein synthesis by increasing the amount of viral gene products made. PMID- 6287222 TI - Isolation and subcloning analysis of functional centromere DNA (CEN11) from Saccharomyces cerevisiae chromosome XI. AB - We have cloned segments of yeast DNA containing the centromere XI-linked MET14 gene. This was done by selecting directly in Saccharomyces cerevisiae for complementation of a met14 mutation after transformation with a hybrid plasmid DNA genomic library. Genetic evidence indicates that functional centromere DNA (CEN11) from chromosome XI is also contained on the segment of S. cerevisiae DNA cloned in pYe(MET14)2. This plasmid is maintained stably in budding S. cerevisiae cultures and segregates predominantly 2+:20- through meiosis. The CEN11 element has been subcloned in vector YRp7' on an S. cerevisiae DNA fragment 900 base pairs in length [pYe(CEN11)10]. The mitotic and meiotic behavior of plasmids containing CEN11 plus a DNA replicator (ars) indicates that the centromere DNA sequences enable these plasmids to function as true minichromosomes in S. cerevisiae. PMID- 6287223 TI - Syntheses and stabilities of proteins related to the polyoma small T antigen in Escherichia coli. AB - We compared the syntheses and turnovers of two proteins related to the polyoma small T antigen synthesized in Escherichia coli from plasmids containing polyoma genomic segments joined to lac control elements. A protein with an authentic polyoma N terminus was more unstable than a protein with N-terminal amino acids derived from beta-galactosidase. Both were more unstable than most bacterial proteins. PMID- 6287224 TI - Direct demonstration of genetic alterations at the dihydrofolate reductase locus after gamma irradiation. AB - Gamma ray-induced mutants of Chinese hamster ovary cells lacking dihydrofolate reductase activity were screened for DNA sequence changes at the locus specifying this activity by using a cloned cDNA probe. Two of nine mutants screened displayed an altered restriction fragment pattern suggesting the occurrence of DNA deletions or rearrangements. PMID- 6287226 TI - Phosphorylation of a 36,000 Mr cellular protein in cells infected with partial transformation mutants of rous sarcoma virus. AB - We have isolated and characterized mutants of Rous sarcoma virus which induce some parameters of transformation but fail to fully induce other parameters. We believe these mutants code for a pp60src which phosphorylates some targets well but phosphorylates others poorly. Using these mutants, we examined the phosphorylation of a 36,000 Mr protein which is phosphorylated on a tyrosine in cells transformed by Rous sarcoma virus, in an attempt to correlate this phosphorylation with the expression of specific transformation parameters. We found that phosphorylation of the 36,000 Mr protein was neither necessary nor sufficient for loss of fibronectin or for loss of density-dependent inhibition of growth. Phosphorylation of the protein was not sufficient for morphological alterations, increased hexose transport, or loss of adhesiveness. For the parameters measured, the best correlation was with increased plasminogen activator. In addition, it is noteworthy that cells infected with the mutant CU2 displayed low levels of phosphorylation of the 36,000 Mr protein and also were deficient in anchorage-independent growth and tumorigenicity, raising the possibility that the phosphorylation of the 35,000 Mr protein may be required for malignant growth properties. PMID- 6287225 TI - Structure of the Schizosaccharomyces pombe cytochrome c gene. AB - The cytochrome c gene of the fission yeast Schizosaccharomyces pombe has been cloned by using the Saccharomyces cerevisiae iso-1-cytochrome c gene as a molecular hybridization probe. The DNA sequence and the 5' termini of the mRNA transcripts of the gene have been determined. The DNA sequence has confirmed, with two exceptions, the previously determined protein sequence. The nonrandom distribution of silent third base differences which was observed between the two cytochrome c genes of S. cerevisiae does not extend to the S. pombe cytochrome c gene, suggesting that there are no constraints other than protein function and codon usage which have acted to conserve the cytochrome DNA sequences of the two yeasts. Introduction of the S. pombe cytochrome c gene on a yeast plasmid into a S. cerevisiae mutant which lacked functional cytochrome c transformed that recipient strain for the ability to grow on a nonfermentable carbon source. This implies that the S. pombe cytochrome c gene has all the regulatory signals which are required for its expression in S. cerevisiae, and that none of the amino acid differences between the cytochrome c proteins of the two yeasts has a drastic effect on the function of the protein in vivo. PMID- 6287227 TI - High-efficiency cloning of full-length cDNA. AB - A widely recognized difficulty of presently used methods for cDNA cloning is obtaining cDNA segments that contain the entire nucleotide sequence of the corresponding mRNA. The cloning procedure described here mitigates this shortcoming. Of the 10(5) plasmid-cDNA recombinants obtained per microgram of rabbit reticulocyte mRNA, about 10% contained a complete alpha- of beta-globin mRNA sequence, and at least 30 to 50%, but very likely more, contained the entire globin coding regions. We attribute the high efficiency of cloning full- or nearly full-length cDNA to (i) the fact that the plasmid DNA vector itself serves as the primer for first- and second-strand cDNA synthesis, (ii) the lack of any nuclease treatment of the products, and (iii) the fact that one of the steps in the procedure results in preferential cloning of recombinants with full-length cDNA's over those with truncated cDNA's. PMID- 6287228 TI - Transfer of nonselectable genes into mouse teratocarcinoma cells and transcription of the transferred human beta-globin gene. AB - Teratocarcinoma (TCC) stem cells can function as vehicles for the introduction of specific recombinant genes into mice. Because most genes do not code for a selectable marker, we investigated the transformation efficiency of vectors with a linked selectable gene. In one series, TCC cells first selected for thymidine kinase deficiency were treated with DNA from the plasmid vector PtkH beta 1 containing the human genomic beta-globin gene and the thymidine kinase gene of herpes simplex virus. A high transformation frequency was obtained after selection in hypoxanthine-aminopterin-thymidine medium. Hybridization tests revealed that the majority of transformants had intact copies of the human gene among three to six total copies per cell. These were associated with cellular DNA sequences as judged from the presence of additional new restriction fragments and from stability of the sequences in tumors produced by injecting the cells subcutaneously. Total polyadenylate-containing RNA from cell cultures of two out of four transformants examined showed hybridization to the human gene probe: one RNA species resembled mature human beta-globin mRNA transcripts; the others were of larger size. In differentiating tumors, various tissues, including hematopoietic cells of TCC provenance could be found. In a second model set of experiments, wild-type TCC cells were used to test a dominant-selection scheme with pSV-gpt vectors. Numerous transformants were isolated, and their transfected DNA was apparently stably integrated. Thus, any gene of choice can be transferred into TCC stem cells even without mutagenesis of the cells, and selected cell clones can be characterized. Cells of interest may then be introduced into early embryos to produce new mouse strains with predetermined genetic changes. PMID- 6287229 TI - Tyrosine phosphorylation of a 50K cellular polypeptide associated with the Rous sarcoma virus transforming protein pp60src. AB - We have examined the phosphorylation of a 50,000-dalton cellular polypeptide associated with the Rous sarcoma virus (FSV) transforming protein pp60-src. It has been shown that pp60src forms a complex with two cellular polypeptides, an 89,000-dalton heat-shock protein (89K) and a 50,000-dalton phosphoprotein (50K). The pp60src-associated protein kinase activity phosphorylates at tyrosine residues, and the 50K polypeptide present in the complex contains phosphotyrosine and phosphoserine. These observations suggest that the 50K polypeptide may be a substrate for the protein kinase activity of pp60src. To examine this possibility, we isolated the 50K polypeptide by two-dimensional polyacrylamide gel electrophoresis from lysates of uninfected or virally infected cells. Tryptic phosphopeptide analysis indicated that the 50K polypeptide isolated by this method was the same polypeptide as that complexed to pp60src. In uninfected cells or cells infected by a transformation-defective mutant, the 50K polypeptide contained phosphoserine but little or no phosphotyrosine. In cells infected by Schmidt-Ruppin or Prague RSV, there was a 40- to 50-fold increase in the quantity of phosphotyrosine in the 50K protein. Thus, the phosphorylation of the 50K polypeptide at tyrosine is dependent on the presence of pp60src. However, the 50K polypeptide isolated from cells infected by temperature-sensitive mutants of RSV was found to be phosphorylated at tyrosine at both permissive and nonpermissive temperatures; this behavior is different from that of other substrates or putative substrates of the pp60src kinase activity. It is possible that the 50K polypeptide is a high-affinity substrate of pp60src. PMID- 6287230 TI - Accessibility of ribosomal genes to trimethyl psoralen in nuclei of Physarum polycephalum. AB - We have probed the accessibility of the genes for rRNA in Physarum polycephalum by using the photoreactive DNA cross-linking agent 4,5',8-trimethyl psoralen. Nuclei isolated from actively growing Physarum were treated with trimethyl psoralen and irradiated with 360-nm light in order to form cross-links. The palindromic, extrachromosomal rDNA then was isolated, and the positions of cross links were determined by electron microscopy of the DNA under totally denaturing conditions. The results indicate that the frequency of cross-linking, after correction for base sequence bias of the reaction, is up to sixfold higher in the transcribed regions than in the central or the terminal spacer regions. There is no detectable heterogeneity among the different rDNA molecules or between the halves of a single molecule. Cross-linked molecules invariably occur in a linear as opposed to a cruciform structure. The preferential cross-linking of the transcribed region is nearly eliminated in spherules, a dormant transcriptionally inactive form in the Physarum life cycle. PMID- 6287231 TI - Construction, replication, and chromatin structure of TRP1 RI circle, a multiple copy synthetic plasmid derived from Saccharomyces cerevisiae chromosomal DNA. AB - Transformation studies with Saccharomyces cerevisiae (bakers' yeast) have identified DNA sequences which permit extrachromosomal maintenance of recombinant DNA plasmids in transformed cells. It has been hypothesized that such sequences (called ARS for autonomously replicating sequence) serve as initiation sites for DNA replication in recombinant DNA plasmids and that they represent the normal sites for initiation of replication in yeast chromosomal DNA. We have constructed a novel plasmid called TRP1 R1 Circle which consists solely of 1,453 base pairs of yeast chromosomal DNA. TRP1 RI Circle contains both the TRP1 gene and a sequence called ARS1. This plasmid is found in 100 to 200 copies per cell and is relatively stable during both mitotic and meiotic cell cycles. Replication of TRP1 RI Circle requires the products of the same genes (CDC28, CDC4, CDC7, and CDC8) required for replication of chromosomaL DNA. Like chromosomal DNA, its replication does not occur in cells arrested in the B1 phase of the cell cycle by incubation with the yeast pheromone alpha-factor. In addition, TRP1 RI Circle DNA is organized into nucleosomes whose size and spacing are indistinguishable from that of bulk yeast chromatin. These results indicate that TRP1 RI Circle has the replicative and structural properties expected for an origin of replication from yeast chromosomal DNA. Thus, this plasmid is a suitable model for further studies of yeast DNA replication in both cells and cell-free extracts. PMID- 6287232 TI - Chicken ovalbumin gene fused to a herpes simplex virus alpha promoter and linked to a thymidine kinase gene is regulated like a viral gene. AB - We are describing a system for the introduction, selection, and expression of eucaryotic genes in higher eucaryotic cells. The carrier consisted of the herpes simplex virus 1 (HSV-1) tk gene covalently linked to an HSV-1 alpha promoter directed away from the tk gene. In this study we fused to the alpha promoter the 5' transcribed noncoding sequences and the coding sequences of the chicken oviduct ovalbumin gene. Cells converted to the TK+ phenotype with this chimeric fragment produced an ovalbumin precursor which was processed and secreted into the extracellular fluid. The ovalbumin gene utilized the HSV-1 alpha promoter and was regulated as a viral gene inasmuch as inversion of the genomic DNA relative to the alpha promoter resulted in no ovalbumin synthesis, and production of ovalbumin was enhanced after superinfection with HSV-1. Synthesis of ovalbumin was not detected when cDNA was linked to the HSV-1 alpha promoter. The carrier system described in this study is suitable for introduction, selection, and expression of eucaryotic genes whose natural promoter is either weak or requires the presence of regulatory elements which may be absent from undifferentiated cells in culture. PMID- 6287234 TI - The response of chicken embryo dermal fibroblasts to cytochalasin B is altered by Rous sarcoma virus-induced cell transformation. AB - The drug cytochalasin B (CB), which disrupts the cellular microfilament network, allows the identification of as yet unclassified structural differences between normal and Rous sarcoma virus-transformed chicken embryo fibroblasts. When exposed to CB, normal chick fibroblasts attain an arborized or dendritic morphology. This results as the cytoplasm collapses upon the remaining structural and adhesive components of the cell. Rous sarcoma virus-transformed cells did not form or maintain these dendritic-like processes in the presence of CB and, as a result, rounded up but still remained attached to the substrate. With a temperature-sensitive mutant of Rous sarcoma virus, LA24A, it was possible to show that these effects are completely reversible and dependent on the expression of pp60src. The cytoskeleton in these CB-treated cells was examined by both immunofluorescence and electron microscopy. After exposure to CB, the microfilaments were found to be disrupted similarly throughout both the transformed and the nontransformed cells. In the nontransformed cells arborized by exposure to CB, the extended processes were found to contain intermediate filaments in an unusually high concentration and degree of organization. The distribution of these filaments in the central body of the arborized cells was random. This lower concentration and random distribution was similar to that seen throughout the transformed cells rounded up by exposure to CB. The failure of these transformed cells to arborize in CB indicates that the structural component(s) which is necessary for the formation or maintenance or both of the arborized state is altered by the expression of pp60src. PMID- 6287233 TI - Overproduction of dihydrofolate reductase and gene amplification in methotrexate resistant Chinese hamster ovary cells. AB - Stable isolates of Chinese hamster ovary cells that are highly resistant to methotrexate have been selected in a multistep selection process. Quantitative immunoprecipitations have indicated that these isolates synthesize dihydrofolate reductase at an elevated rate over its synthesis in sensitive cells. Restriction enzyme and Southern blot analyses with a murine reductase cDNA probe indicate that the highly resistant isolates contain amplifications of the dihydrofolate reductase gene number. Depending upon the parenteral line used to select these resistant cells, they overproduce either a wild-type enzyme or a structurally altered enzyme. Karyotype analysis shows that some of these isolates contain chromosomes with homogeneously staining regions whereas others do not contain such chromosomes. PMID- 6287235 TI - Half-life of the Rous sarcoma virus transforming protein pp60src and its associated kinase activity. AB - The half-life of metabolically labeled pp60src of the Prague A strain of Rous sarcoma virus and of several transformation-defective, temperature-sensitive mutants was investigated by pulse-labeling infected cells with [35S]methionine, chasing for different times, and immunoprecipitating pp60src with tumor-bearing rabbit serum. These experiments showed that pp60src has a short half-life of approximately 60 min under normal physiological conditions and that the mutant pp60src proteins have similar half-lives to the wild type, irrespective of whether the cells are kept at the nonpermissive (42 degrees C) or permissive (35 degrees C) temperature. The half-life of the pp60src -associated kinase activity was determined by monitoring its decay by the immunoglobulin G heavy chain assay after the cells had been treated with several inhibitors of protein synthesis. In these experiments the kinase half-life was much longer than expected from the half-life of pp60src. The apparent contradiction between the half-lives of the kinase activity and the [35S]methionine-labeled pp60src protein could be resolved by the observation that treatment of cells with inhibitors of protein synthesis stabilized pp60src, resulting in a greatly extended half-life. Inhibitors of protein synthesis also extended the half-life of the gag precursor polypeptide, Pr76, suggesting that a host factor(s) may be required for the efficient intracellular processing of this polypeptide to the gag proteins. PMID- 6287236 TI - Selection and characterization of a glucokinase-deficient mutant of Tetrahymena thermophila. AB - We have isolated a mutant of Tetrahymena thermophila that is resistant to inhibition of growth by the glucose analog 2-deoxyglucose. The mutant exhibits a deficiency in a cytoplasmic glucokinase. This enzymatic defect and the attendant inability to convert 2-deoxyglucose to toxic phosphorylated derivatives is apparently the sole basis for the mutant phenotype since transport of glucose and 2-deoxyglucose is unimpaired; there is no elevation of glucose-6-phosphatase activity, which could decrease the level of toxic 2-deoxyglucose metabolites. Genetic analyses have shown that the mutant allele is recessive and inherited as a single Mendelian mutation. The glucokinase-deficient strain described here is useful for the selection of other mutants in this organism and for the investigation of various cellular processes initiated or modulated by glucose and its analogs. We have exploited the molecular defect in this strain to investigate the initial steps in the cyclic AMP-mediated repression of galactokinase gene expression which is caused by glucose. PMID- 6287237 TI - Preliminary characterization of the transcriptional and translational products of the Saccharomyces cerevisiae cell division cycle gene CDC28. AB - The CDC28 gene was subcloned from a plasmid containing a 6.5-kilobase-pair segment of Saccharomyces cerevisiae DNA YRp7(CDC28-3) by partial digestion with Sau3A and insertion of the resulting fragments into the BamHI sites of YRp7 and pRC1. Recombinant plasmids were obtained containing inserts of 4.4 and 3.1 kilobase pairs which were capable of complementing a cdc28(ts) mutation. R-loop analysis indicated that each yeast insert contained two RNA coding regions of about 0.8 and 1.0 kilobase pairs, respectively. In vitro mutagenesis experiments suggested that the smaller coding region corresponded to the CDC28 gene. When cellular polyadenylic acid-containing RNA, separated by agarose gel electrophoresis after denaturation with glyoxal and transferred to nitrocellulose membrane, was reacted with labeled DNA from the smaller coding region, and RNA species of about 1 kilobase in length was detected. Presumably, the discrepancy in size between the R-loop and electrophoretic determinations is due to a segment of polyadenylic acid which is excluded from the R-loops. By using hybridization of the histone H2B mRNAs to an appropriate probe as a previously determined standards, it was possible to estimate the number of CDC28 mRNA copies per haploid cell as between 6 and 12 molecules. Hybrid release translation performed on the CDC29 mRNA directed the synthesis of a polypeptide of 27,000 daltons, as determined by polyacrylamide gel electrophoresis in sodium dodecyl sulfate. This polypeptide was not synthesized when mRNA prepared from a cdc28 nonsense mutant was translated in a parallel fashion. However, if the RNA from a cell containing the CDC28 gene on a plasmid maintained at a high copy number was translated, the amount of in vitro product was amplified fivefold. PMID- 6287238 TI - Isolation of the thymidylate synthetase gene (TMP1) by complementation in Saccharomyces cerevisiae. AB - The structural gene (TMP1) for yeast thymidylate synthetase (thymidylate synthase; EC 2.1.1.45) was isolated from a chimeric plasmid bank by genetic complementation in Saccharomyces cerevisiae. Retransformation of the dTMP auxotroph GY712 and a temperature-sensitive mutant (cdc21) with purified plasmid (pTL1) yielded Tmp+ transformants at high frequency. In addition, the plasmid was tested for the ability to complement a bacterial thyA mutant that lacks functional thymidylate synthetase. Although it was not possible to select Thy+ transformants directly, it was found that all pTL1 transformants were phenotypically Thy+ after several generations of growth in nonselective conditions. Thus, yeast thymidylate synthetase is biologically active in Escherichia coli. Thymidylate synthetase was assayed in yeast cell lysates by high-pressure liquid chromatography to monitor the conversion of [6-3H]dUMP to [6 3H]dTMP. In protein extracts from the thymidylate auxotroph (tmp1-6) enzymatic conversion of dUMP to dTMP was barely detectable. Lysates of pTL1 transformants of this strain, however, had thymidylate synthetase activity that was comparable to that of the wild-type strain. PMID- 6287239 TI - Regulation of the cellular p53 tumor antigen in teratocarcinoma cells and their differentiated progeny. AB - F9 embryonal carcinoma cells express high levels of a 53,000-molecular-weight cellular tumor antigen called p53. When F9 cell cultures are treated with retinoic acid and dibutyryl adenosine 3',5'-phosphate, they differentiate, predominantly into endoderm-like cells. This differentiation is accompanied by a marked decrease in the levels of p53. The mechanism(s) responsible for this decline in the level of p53 in differentiated cells was investigated. The results demonstrate that the high levels of p53 in F9 cells relative to their differentiated progeny were not due to alterations in the stability or turnover of this protein. Rather, the regulation during differentiation involved a marked decrease in the amount of in vitro translatable p53 mRNA detected in the differentiated cell cultures. This mechanism is unlike the one operating during the simian virus 40 infection or transformation, where the increased levels of p53 are largely due to the increased stability of the p53 protein. PMID- 6287240 TI - Inhibition of phorbol ester-receptor binding by a factor from human serum. AB - The inhibition of receptor binding of [3H]phorbol-12,13-dibutyrate (PDBu) by a factor from human serum was characterized. The serum factor inhibited [3H]PDBu binding in intact monolayer cultures of the rat embryo cell line CREF N and in a subcellular system containing membranes from these cells. Inhibition occurred at both 37 and 4 degrees C and was rapid and reversible. An analysis of [3H]PDBu binding in the presence of the serum factor indicated that inhibition of [3H]PDBu binding by the serum factor was noncompetitive. Using gel filtration to separate the serum factor from free [3H]PDBu, we obtained evidence that the serum factor does not act by binding or trapping the [3H]PDBu. Unlike the phorbol ester tumor promoters, the serum factor alone did not stimulate the release of choline or arachidonic acid from cellular phospholipids, nor did it inhibit the binding of 125I-labeled epidermal growth factor to cellular receptors. The factor did, however, antagonize the inhibition of epidermal growth factor binding induced by PDBu. Sera from pregnant women were, in general, more inhibitory of [3H]PDBu binding than were those from nonpregnant women, which were more inhibitory than those from men. During these studies we found that CREF N cells responded to being grown in the presence of PDBu by partial down regulation of the phorboid receptor. The 50% effective dose for down regulation was 8 nM PDBu, and the maximum effect occurred after 6 h. Taken together, our results indicate that the serum factor inhibits [3H]PDBu binding by a direct physical effect at the level of the phorboid receptors or their associated membranes. It would appear that if this factor acts in vivo, then it might antagonize certain effects of this class of tumor promoters. PMID- 6287241 TI - Comparison of phenotypic expression with genotypic transformation by using cloned, selectable markers. AB - The frequency of phenotypic expression of the herpes simplex virus type 1 tk and Escherichia coli gpt genes was compared with the frequency of genotypic transformation after calcium phosphate-mediated DNA transfection of a number of tk- and hprt- cell lines. In three of the five lines tested, the frequency of phenotypic expression was at most 10-fold higher than that of genotypic transformation as indicated by frequency of HAT resistance. The remaining two lines showed phenotypic responses which were 50- to 100-fold greater than the genotypic responses. The data indicate that the efficiency of DNA-mediated transformation with some cell lines can be limited by events after the uptake and expression of transfected DNA. PMID- 6287242 TI - Kinetics of activation of the first component of complement (C1) by IgG oligomers. AB - The rate of activation of the first component of complement, Cl, by IgG oligomers was investigated. The kinetics of Clr activation exhibited a pronounced lag phase at low IgG concentration and were followed by a rapid conversion of the proenzyme Cls to Cls. Activation of Cl resulted in a conformational change of this complex. Bases on these results, a model of Cl activation by IgG oligomers is proposed: The efficiency of Cl activation by IgG oligomers is in parallel to their binding affinity to Clq. Clr undergoes a slow rearrangement to a conformation which is not stable and is autoactivated, then Clr rapidly converts Cls to Cls. PMID- 6287243 TI - Dissociation of C1 and concentration dependence of its activation kinetics. AB - The activation of the zymogen C1s to the enzyme C1s in the human C1 complex [C1q(c1rC1s)2] was studied as a function of the concentrations of (C1rC1s)2 and C1q which were saturated with oligomers of rabbit IgG. A large concentration dependence of the sigmoidal kinetics was observed in the 2-180 nM concentration range. This was explained by association-dissociation equilibria between the antibody-saturated C1q and various forms of the (C1rC1s)2 complex (unactivated to activated). The establishment of these equilibria (binding constant 2 x 10(7) M 1) was assumed to be fast as compared to the rates of the activation steps (rate constants 10(-3) and 10(-2) sec-1 at 30 degrees C). The fast re-equilibration of the C1 complex explains the finding that small amounts of antibody-saturated C1q catalysed the activation of large amounts of C1s. The interpretation of the kinetic results was supported by a direct demonstration of the dissociation of C1 into C1q and (C1rC1s)2 by analytical and density gradient centrifugation. No difference was found between the rates of activation and the dissociation properties of reconstituted C1 and C1 isolated from serum. PMID- 6287244 TI - Specificity of rabbit antisera for peptide 81-104 of horse cytochrome c is dominated by C-terminal residues. AB - Sera have been prepared in rabbits against the carboxy terminal cyanogen bromide fragment of horse cytochrome c (81-104) conjugated to bovine serum albumin (BSA). The specificity of these sera has been studied by radioimmunoassay and reveals that only peptide 81-104 binds the antibody. Neither whole cytochrome c nor peptides 1-65 and 66-80 displace 125I-81-104 from specific antibody, while cold peptides 81-104 is active in this regard, as expected. Fine specificity studies with isolated heterologous peptides 81-104 from the bovine, rabbit, canine, chicken, pigeon and tuna species of cytochrome c reveal that a major antigenic determinant is influenced by sequence positions 100 and 103 while a minor antigenic determinant is influenced by residue 92. These results are exactly mirrored and supported by inhibition studies with the isolated chymotryptic fragments 98-104 and 83-97. Studies with tryptic, chymotryptic and peptic digests of horse cytochrome c reveal that these natural proteases can generate peptides with the appropriate peptide sequences to react with antibodies produced against the peptide 81-104-BSA conjugate. PMID- 6287245 TI - [Infectious gastroenteritis in childhood (author's transl)]. AB - Stool specimens from 126 children with acute gastroenteritis and from 42 controls were screened bacteriologically and virologically including electron-microscopy. Rota viruses were found in 36.5% of the children with diarrhoea, enteropathogenic E. coli in 13.5% and candida in 7.1%. These pathogens were not found in the controls. Enteroviruses were found in almost the same frequency in both groups (13.5% in children with gastroenteritis, 16.6% in the controls). The etiologic role of adenoviruses, which could be seen in 5.6% of the diarrhoea group, remains unexplained. PMID- 6287246 TI - Malignant fibrothecoma: pathomorphological comments. PMID- 6287247 TI - [Clinical aspects of synoviomas (author's transl)]. PMID- 6287248 TI - Mutagenicity of some methylated benzo[a]pyrene derivatives. AB - The mutagenicity of benzo[a]pyrene (BP) and a number of methylated derivatives towards Salmonella typhimurium has been tested. The most mutagenic derivative tested was 6-methylbenzo[a]pyrene which produced about twice the number of revertants as did BP, 11-Methylbenzo[a]pyrene was slightly more mutagenic than BP. All the other compounds tested (7-, 8-, 9- and 10-methylbenzo[a]pyrene and 7,8- and 7,10-dimethylbenzo[a]pyrene) were significantly less active than benzo[a]pyrene. With the exception of 6-methylbenzo[a]pyrene, these results closely parallel the known carcinogenicity of the methylated benzo[a]pyrenes, and support the view that metabolic activation of BP may involve the 7-10 positions which are blocked in the methylated compounds. PMID- 6287249 TI - Induction of sister-chromatid exchanges in human lymphocytes by extracts of particulate emissions from a diesel engine. AB - Particulate matter was collected from the emissions of a diesel-powered engine and the organic components were extracted. The extract induced a dose-related increase in sister-chromatid exchanges in human lymphocytes with a potency of about one-fifth that of benzo[a]pyrene. PMID- 6287250 TI - 3-Aminobenzamide synergistically increases sister-chromatid exchanges in cells exposed to methyl methanesulfonate but not to ultraviolet light. AB - 3-Aminobenzamide, an inhibitor of poly(ADP-ribose) synthesis, increased baseline sister-chromatid exchange (SCE) frequencies and acted synergistically with the alkylating agent methyl methanesulfonate to induce exchanges in Chinese hamster ovary and SV40-transformed human (GM637) cells. In contrast, 3-aminobenzamide did not affect the frequency of ultraviolet light-induced SCEs. Our data suggest that, in these 2 cell types, synthesis of poly(ADP-ribose) is more important in damage and repair after exposure to an alkylating agent than after exposure to ultraviolet light. PMID- 6287251 TI - Mutagenicity of benzo[a]pyrene and the antioxidant phenol at the HGPRT locus of V79 chinese hamster cells. PMID- 6287252 TI - Survival of 60Co-irradiated herpes simplex virus in 15 human diploid fibroblast cell strains. AB - The present study was designed to determine the extent to which herpes simplex virus (HSV) may be utilized to study the repair of DNA damaged by ionizing radiation. We investigated the survival of 60Co-irradiated HSV in cell strains derived from 2 normal controls and 13 patients with a broad range of diseases associated with possible DNA repair deficiencies. Irradiation was performed under two conditions to vary the type of damage incurred by the virus. HSV survival was greatly enhanced when the virus was irradiated in such a way that the indirect effects of ionizing radiation were minimized. We found no correlation between cellular hypersensitivity to ionizing radiation and survival of irradiated HSV. Reduced levels of virus survival were found in only 1 cell strain. When cells were treated with ionizing radiation or UV light prior to infection, no enhancement of virus survival was observed. PMID- 6287253 TI - Metabolic activation of benzo[a]pyrene, aflatoxin B1, and dimethylnitrosamine by a human hepatoma cell line. AB - The metabolism of chemical carcinogens by a human hepatoma cell line, huH-1, was studied. The huH-1 line has been derived from a hepatoma of a 57-year-old HBs antigen carrier and cultivated for several years. The hepatoma cells metabolized about 90% of 5 microM benzo[a]pyrene into water-soluble products within 24 h. Aryl hydrocarbon hydroxylase activity in huH-1 cells was induced to 24 times higher than the basal level by treatment with 13 microM benz[a]anthracene for 24 h. Metabolic activation of benzo[a]pyrene, dimethylnitrosamine and aflatoxin B1 by huH-1 cells was observed by cell-mediated sister-chromatid exchange assay. Sister-chromatid exchanges in human diploid fibroblasts were observed in the cultures mixed with or without huH-1 cells. All 3 chemicals induced sister chromatid exchanges in human fibroblasts far more efficiently in the cultures mixed with huH-1 cells than in those without huH-1 cells. Some characteristics of huH-1 cells as a human cell-mediated metabolic activation system for carcinogens are discussed. PMID- 6287254 TI - Effects of benzo[a]pyrene diol epoxide adducts on DNA synthesis in mammalian cells. AB - The replication of DNA containing anti-benzo[a]pyrene diol epoxide (BPDE) adducts was studied in mammalian cells by first treating SV40 virus with BPDE in vitro, then infecting cells with virus containing a known number of adducts in the DNA. Viral transcription products necessary for replication were supplied by co infection with an untreated virus containing a deletion as a DNA marker. Thus, only replicative effects of BPDE adducts were manifested. Delayed replication of the DNA from BPDE-treated virus, relative to the DNA containing the deletion, was observed, but in time most or all of the infecting molecules were able to replicate. The results are consistent with the hypothesis that adducts of BPDE in DNA block DNA synthesis in vivo, as they do in vitro, and that the block is gradually overcome by a repair mechanism that eliminates the adducts responsible for blockage or by delayed replicative bypass of the adducts. In spite of the ability of the system to overcome the delay in replication, the viability of the BPDE-treated virus in plaque assay was low, suggesting a persistent defect in transcription or a high level of error in repair or bypass replication. PMID- 6287255 TI - Host cell reactivation by excision repair is error-free in human cells. AB - Do host cell repair processes affect the mutagenesis of UV-irradiated virus in human cells? The answer was obtained by investigating the mutagenesis of UV irradiated herpes simplex virus after the irradiated virus was grown in human cells that possess normal repair capacity (normal) or lack excision repair (XPA) or post-replication repair (XP var). Evidence is presented which indicate that XPA cells express no host cell reactivation, while XP var cells express the normal level. Viral mutagenesis was measured as the fraction of the progeny of the surviving virus capable of plaque formation in the presence of iododeoxycytidine. In the normal and XPA cells mutagenesis of the irradiated virus increased linearly with UV exposure. The UV exposure needed to yield a given mutagenesis level for virus grown in XPA cells was much lower than that for virus grown in normal cells. However, when the mutation frequencies were compared at similar virus survival levels, the data from virus grown in normal cells and in XPA cells were indistinguishable. Mutagenesis in XP var cells increased as dose squared and was similar in magnitude to that in normal cells. Thus the excision repair of normal cells which provided host cell reactivation by removing lethal UV damage also removed mutagenic lesions from the virus with the same efficiency, while the repair deficiency of XP var cells had a minor role in host cell reactivation and in mutagenesis. This demonstrates that in human cells host cell reactivation by excision repair is primarily an error-free process. PMID- 6287256 TI - Mechanisms for chromosomal aberrations in mammalian cells. AB - Experimental evidence is presented for the involvement of DNA double-strand breaks in the formation of radiation-induced chromosomal aberrations. When X irradiated cells were post-treated with single-strand specific Neurospora crassa endonuclease (NE), the frequencies of all classes of aberration increased by about a factor 2. Under these conditions, the frequencies of DNA double-strand breaks induced by X-rays (as determined by neutral sucrose-gradient centrifugation), also increased by a factor of 2. The frequency of chromosomal aberrations induced by fast neutrons (which predominantly induce DNA double strand breaks) was not influenced by post-treatment with NE. Inhibition of poly(ADP-ribose) polymerase, an enzyme that uses DNA with double-strand breaks as an optimal template, by 3-aminobenzamide also increased the frequencies of X-ray induced chromosomal aberrations, which supports the idea that DNA double-strand breaks are important lesions for the production of chromosomal aberrations induced by ionizing radiation. PMID- 6287257 TI - The maxicircle of Trypanosoma brucei kinetoplast DNA hybridizes with a mitochondrial gene encoding cytochrome oxidase subunit II. AB - A restriction endonuclease fragment of the maxicircle of Trypanosoma brucei brucei kinetoplast DNA hybridizes with a cloned mitochondrial DNA sequence which encodes cytochrome oxidase subunit II of Zea mays. A cloned mitochondrial DNA sequence encoding cytochrome oxidase subunit II of Saccharomyces cerevisiae also hybridized with kDNA, but exhibits less homology with the maxicircle than does the maize gene. The hybridizing maxicircle DNA was localized to a 2.8 kbp segment which is bounded by TaqI restriction endonuclease sites and nearby HindIII and EcoRI restriction sites. The TaqI restriction fragment is conserved between T. brucei brucei, T. brucei rhodesiense and T. brucei gambiense and hybridizes with the Zea mays probe in each case. PMID- 6287259 TI - Acyclovir-resistant herpes simplex virus. PMID- 6287258 TI - Pseudohypoparathyroidism: the molecular basis for hormone resistance--a retrospective. PMID- 6287260 TI - Defect in ACTH control persisting after removal of an ectopic source of ACTH. PMID- 6287261 TI - Presence of delta9-tetrahydrocannabinol in human milk. PMID- 6287262 TI - Hepatoblastoma in an infant born to a mother after hormonal treatment for sterility. PMID- 6287264 TI - Epstein-Barr virus-induced B-cell lymphoma after renal transplantation. PMID- 6287263 TI - Anticipatory nausea and vomiting associated with cancer chemotherapy. PMID- 6287265 TI - MMWR feels the budgetary crunch. PMID- 6287266 TI - MMWR feels the budgetary crunch. PMID- 6287267 TI - Congenital cytomegalovirus infection. PMID- 6287268 TI - Cryopreservation of human peripheral blood lymphoid cells reactive to allogeneic antigens, virus-transformed autologous cells and autologous leukemia cells in in vitro sensitization. PMID- 6287269 TI - Mitochondrial mosaics-maturases on the move. PMID- 6287270 TI - Kainic acid stimulates excitatory amino acid neurotransmitter release at presynaptic receptors. PMID- 6287272 TI - A multicopy insertion sequence in the bovine genome with structural homology to the long terminal repeats of retroviruses. PMID- 6287271 TI - An opioid benzodiazepine. PMID- 6287273 TI - Opioids at Cape Cod. PMID- 6287274 TI - Reversion of a promoter deletion in yeast. AB - Promoter function in yeast has been examined by obtaining revertants of a his3 promoter deletion in vivo. Events which provide a new promoter for the his3 gene include insertion of the transposable element Ty1, rearrangements of the plasmid vector, and chromosomal mutations. A role for dicentric chromosomes as a source of the plasmid rearrangements is discussed. PMID- 6287275 TI - Detection of K+ and Cl-channels from calf cardiac sarcolemma in planar lipid bilayer membranes. AB - The ionic currents underlying the cardiac action potential are believed to be much more complex than those in nerve. During the cardiac action potential, various membrane channels control the flow of K+, Na+, Ca2+ and Cl- across the sarcolemma of cardiac muscle cells. Thus, it has become increasingly clear that a detailed knowledge of the mechanisms that activate (or inactivate) heart channels is required to understand cardiac excitability. We report here the use of planar lipid bilayer techniques to detect and characterize K+ and Cl- channels in purified heart sarcolemma membrane vesicles. We have identified four different types of channel on the basis of their selectivity, conductance and gating kinetics. We present in some detail the properties of a K+ channel and a Cl- channel. We have tentatively identified the K+ channel with the ix type of current found in Purkinje, myocardial ventricular and atrial fibres. The chloride channel might be related to the transient chloride current found in Purkinje fibres. PMID- 6287276 TI - Nucleotide sequence of the rat skeletal muscle actin gene. AB - The actins constitute a family of highly conserved proteins found in all eukaryotic cells. Their conservation through a very wide range of taxonomic groups and the existence of tissue-specific isoforms make the actin genes very interesting for the study of the evolution of genes and their controlling elements. On the basis of amino acid sequence data, at least six different mammalian actins have been identified (skeletal muscle, cardiac muscle, two smooth muscle actins and the cytoplasmic beta- and gamma-actins). Rat spleen DNA digested by the EcoRI restriction enzyme contains at least 12 different fragments with actin-like sequences but only one which hybridized, in very stringent conditions, with the skeletal muscle cloned cDNA probe. Here we describe the sequence of the actin gene in that fragment. The nucleotide sequence codes for two amino acids, Met-Cys, preceding the known N-terminal Asp of the mature protein. There are five small introns in the coding region and a large intron in the 5'-untranslated region. Comparison of the structure of the rat skeletal muscle actin gene with available data on actin genes from other organisms shows that while the sequenced actin genes from Drosophila and yeast have introns at different locations, introns located at codons specifying amino acids 41, 121, 204 and 267 have been preserved at least from the echinoderm to the vertebrates. A similar analysis has been done by Davidson. An intron at codon 150 is common to a plant actin gene and the skeletal muscle acting gene. PMID- 6287277 TI - DNA precursors in chemical mutagenesis: a novel application of DNA sequencing. AB - Recently, we have shown that deoxyribonucleoside residues in the cellular DNA precursor pool are generally more susceptible to methylation than are residues within the DNA duplex. The N-1 position of adenosine, for example, was found to be at least 13,000 times more susceptible to methylation by N-methyl-N nitrosourea (MNU) than the same site in the DNA. These results suggest that potential sites for alkylation in the double-strand duplex are relatively inaccessible to direct alkylation in vivo. Many of these sites are probably protected from alkylation not only by their position in the interstices of the DNA helix, but also by further in vivo 'packaging' of the DNA in chromatin. We have now used DNA sequencing to demonstrate the incorporation properties of products of the reaction of MNU with dATP and of deoxy-N4-hydroxycytidine triphosphate during DNA replication in vitro by phage T4 DNA polymerase and the 'Klenow' fragment of Escherichia coli pol I. The results suggest that DNA precursor nucleotides due to their greater availability for alkylation, may offer routes for the introduction of alkylated residues into double-stranded DNA. PMID- 6287278 TI - Specific protein-nucleic acid recognition in ribonuclease T1-2'-guanylic acid complex: an X-ray study. AB - RNase T1 is folded into an alpha-helix of 4.5 turns, covered by a four-strand antiparallel beta-sheet. Specific recognition of 2'-guanylic acid arises from hydrogen bonding between main chain peptide groups and the O-6 and N-1-H of guanine, as well as from stacking of Tyr 45 on guanine. At the active site, Glu 58, His 92 and Arg 77 are involved in phosphodiester hydrolysis. PMID- 6287279 TI - Absence of gonadotropin-releasing hormone receptors in human gonadal tissue. PMID- 6287280 TI - A fragment of the SV40 large T-antigen gene transforms. PMID- 6287282 TI - ATP-dependent unwinding of double helix in closed circular DNA by recA protein of E. coli. PMID- 6287281 TI - Enkephalin opens potassium channels on mammalian central neurones. PMID- 6287283 TI - Poliomyelitis--epidemiology, molecular biology and immunology. PMID- 6287284 TI - Similarity of unitary Ca2+ currents in three different species. AB - Membrane Ca2+ currents are the triggers for numerous cellular activities such as secretions, contraction, and oscillations in neural discharge. Different Ca2+ channels are assumed to subserve the various functions; this does not apply to Na+ currents for which there is thought to be only one type of channel. We have examined single Ca2+ channels in cells from three very different species (bird, snail and rat) using the patch-clamp method, and report here that unitary events consisted of small, brief current pulses that often occurred in bursts and were similar for all three species. Thus, reported differences among various Ca2+ current must have other origins. PMID- 6287285 TI - Single-channel currents in isolated patches of plasma membrane from basal surface of pancreatic acini. AB - Precise localization and characterization of conductance pathways in glandular epithelia have so far proved difficult. The patch-clamp technique for high resolution current recording, which has already been applied successfully to a number of electrically excitable cells, can in principle overcome these difficulties. We now report measurements of single-channel currents from isolated patches of plasma membrane (inside-out) from the baso-lateral surface of collagenase-isolated rat and mouse pancreatic acini. We have identified a cation channel having a conductance of approximately 30 pS and a mean open time in the range 0.3-1 s which is dependent on internal calcium. The single-channel current voltage relationship is linear and the mean open time independent of the membrane potential. These channels may, at least in part, account for the Ca2+-mediated neural and hormonal control of pancreatic acinar membrane conductance, which is probably responsible for the Ca2+-dependent acinar fluid secretion. PMID- 6287286 TI - Prostaglandins modulate macrophage Ia expression. AB - Prostaglandins are important modulators of inflammation and of humoral and cellular immune responses. In order to evaluate a possible mechanism for the regulation of immune responses we have studied the effects of prostaglandins on the expression of I-region-associated (Ia) antigens by macrophages. The expression of these glycoproteins is essential for macrophages to function as antigen-presenting cells during the induction of immune responses. The synthesis and membrane expression of Ia, however, is not a constitutive property of the phagocyte but is under regulation and a positive regulation of this process is exhibited by activated T cells. In contrast, a negative regulation is conspicuously found in the neonate where a product from a young replicating macrophage inhibits the expression of Ia by the mature macrophages. We show here that prostaglandins of the E series (PGE) are potent inhibitors of the expression of Ia-antigens on macrophages and that thromboxane B2 (TXB2) antagonizes the effect of PGE. PMID- 6287287 TI - A transforming gene present in human sarcoma cell lines. AB - Morphological transformation of NIH/3T3 cells by transfection with DNA has been used to identify transforming sequences in human tumours. Transforming activity has been reported for DNAs isolated from bladder, mammary, colon and lung carcinomas, neuroblastoma, lymphoid and myeloid tumours. Each of these tissues seems to contain different transforming sequences except for the colon and lung tumours where the same sequence seems to be involved. We now report that in two different human sarcoma cell lines, a fibrosarcoma and an embryonal rhabdomyosarcoma, the DNAs have transforming activity. The transforming gene is the same in both sarcomas but differs from the activated sequences detected in other tumours. We have also found that the transforming gene has no detectable homology to eight retrovirus oncogenes tested. PMID- 6287288 TI - An adenovirus glycoprotein binds heavy chains of class I transplantation antigens from man and mouse. AB - The successful killing of virus-infected cells by cytotoxic T lymphocytes (CTL) is dependent on the recognition of both a viral product and class I antigens of the major histocompatibility complex (MHC) on the infected cell surface. Whether these two entities are found independently on the cell surface and therefore recognized by two different CTL receptors, or whether they are associated together and can therefore be recognized by a single receptor is not known. The association between an adenovirus-encoded glycoprotein expressed on the cell surface early after infection and class I antigens has been investigated and it has been found that antisera against class I antigens can co-precipitate the antigen and the viral glycoprotein from an adenovirus-transformed cell line from the Hooded-Lister rat strain. We show here by in vitro affinity chromatography and in vivo immunoprecipitation that the viral glycoprotein specifically binds to the heavy chain of class I antigens in both man and mouse. PMID- 6287289 TI - Molecular cloning of the gene for human anti-haemophilic factor IX. AB - A functional deficiency of factor IX, one of the coagulation factors involved in blood clotting, leads to the bleeding disorder known as Christmas disease, or haemophilia B. Both this disease and haemophilia A (factor VIII (C) deficiency) are X chromosome-linked and together occur at a frequency of approximately 1 in 10,000 males. The molecular basis for the functional alteration of factor IX in Christmas disease is not clearly understood. As a first step towards the elucidation of the molecular events involved, we have attempted molecular cloning of the factor IX gene. We used a bovine factor IX cDNA clone, isolated using synthetic oligonucleotides as probes, to screen a cloned human gene library. Here we report the isolation and partial characterization of a lambda recombinant phage containing the human factor IX gene. PMID- 6287290 TI - A transient outward current in a mammalian central neurone blocked by 4 aminopyridine. PMID- 6287291 TI - Loss of intervening sequences in genomic mouse alpha-globin DNA inserted in an infectious retrovirus vector. PMID- 6287292 TI - Left-handed Z-DNA is induced by supercoiling in physiological ionic conditions. AB - In physiological ionic conditions (200 mM NaCl), the (dC-dG)16 and (dC-dG)13 blocks in plasmid pRW751 are in a left-handed state when the negative superhelical density of the plasmid is greater than 0.972. As the salt concentration decreases or when (dmC-dG) sequences are present, less negative supercoiling is required to induce the right- to left-handed DNA transition. Furthermore, the single strand-specific nuclease, S1, recognizes and cleaves aberrant structural features at the junction between neighbouring right- and left handed DNA regions. PMID- 6287293 TI - Corticotropin releasing activity of the new CRF is potentiated several times by vasopressin. AB - Initially the hypothalamic factor responsible for the release of corticotropin (CRF), was thought to be a simple peptide. More recent work has led to the conclusion that CRF is a multifactorial complex. In 1979 we proposed that vasopressin, much disputed as a CRF candidate, was a major constituent of the complex, interacting with a potentiating the CRF activity of the other component(s). The recent characterization of a 41 residue ovine hypothalamic peptide capable of releasing adrenocorticotropic hormone (ACTH) in a dose-related manner has allowed us to compare its CRF bioactivity with that of vasopressin and simple extracts of the hypothalamus, and to investigate any interaction it may have with vasopressin and other hypothalamic factors in the release of ACTH. We report here that the new CRF is more potent than vasopressin in releasing ACTH. When given simultaneously with vasopressin a fourfold potentiation of CRF activity with steep dose-response characteristics were observed. It also potentiated vasopressin-free hypothalamic extracts, suggesting that a new CRF does not account for all the nonvasopressin portion of the CRF complex. PMID- 6287294 TI - A Cl- conductance activated by hyperpolarization in Aplysia neurones. AB - Although many voltage-gated cation channels have been described and extensively studied in biological membranes, there are very few examples of voltage-gated anion channels. Chloride conductances activated by depolarization have been observed in skate electroplaque and in frog and chick skeletal muscle. A Cl- conductance activated by hyperpolarization has been suggested both for frog muscle treated with acid (pH 5) solutions, and for crayfish muscle where it could account for the fact that the pronounced inward-going rectification of the I-V curve disappears if the fibres have been soaked in a Cl(-)-free solution. More recently, voltage-dependent anion channels extracted from biological membranes have been incorporated into artificial membranes. I now report that in Aplysia neurones, and in particular those in which the internal Cl- concentration has been increased, a Cl- conductance can be observed which is slowly activated by hyperpolarization and shows a vary steep voltage dependence. This time- and voltage-dependent Cl- conductance probably exists also in many other cells. Its presence might explain why it is difficult when using KCl-filled microelectrodes to maintain prolonged hyperpolarizations. This Cl- conductance constitutes a new type of inward-going rectification distinct both from the classical "anomalous rectification' which involves selective K+ channels and from the current termed if in heart muscle that is presently attributed to a cationic conductance. PMID- 6287295 TI - Unique cell lines harbouring both Epstein-Barr virus and adult T-cell leukaemia virus, established from leukaemia patients. AB - Members of three distinct classes of animal virus have been associated with naturally occurring neoplasias in man: Epstein--Barr virus (EBV), a DNA virus belonging to herpesvirus group, papillomavirus, and a novel human RNA (retro) virus, human T-cell leukemia virus or adult T-cell leukaemia (ATL) virus. We have established seven continuous cell lines from ATL patients and 0.1-7% of these cells consistently express ATL-specific antigens (ATLA). EBV-associated nuclear antigen (EBNA) is also found in more than 90% of these cells. We have cloned cells from two of these lines and show here that both ATLA and EBNA were present in the same B-cell clone carrying surface immunoglobulin (sIg). PMID- 6287296 TI - T4 late transcripts are initiated near a conserved DNA sequence. AB - Bacteriophage T4 late transcription is unusual, among prokaryotes, in its complexity. Late transcription requires the host RNA polymerase, the products of T4 genes, 33, 45 and 55, and other small polypeptides, the genes of which have not been identified. In addition the DNA template must be "competent' for late transcription. First the DNA must contain the substituted base 5-hydroxymethyl cytosine in place of cytosine (this requirement is eliminated by a mutation in the T4 alc gene). Second, the DNA must be replicating, although late transcription can be uncoupled from DNA replication by mutations in the T4 genes coding for DNA ligase (gene 30) and DNA exonuclease (gene 46). We report here the location of the initiation sites of the messenger RNAs (mRNAs) synthesized in vivo from four late genes (genes 21, 22, 23 and 36) by S1 nuclease mapping and we have determined the DNA sequences at these sites. We have found strong homology to the sequence TATAAATACTATT immediately upstream from the 5' ends of the late messages and we suggest that this sequence is specifically recognized by the complex responsible for late transcription. Also, we have examine gene 23 mRNA synthetized in the absence of DNA replication using the 30- 46- mutant described above and find that it is identical to the true late transcript synthesized in normal infections. PMID- 6287297 TI - Function of cyclic GMP in acetylcholine-induced contraction of coronary smooth muscle. PMID- 6287298 TI - Adenosine receptor agonists: binding and adenylate cyclase stimulation in rat liver plasma membranes. AB - N6-Cyclohexyl[3H]adenosine([3H]CHA,[3H]adenosine, and 5'N ethylcarboxamide[3H]adenosine ([3H]NECA), potent agonists in adenosine-responsive cellular systems, have been used to identify adenosine binding sites in rat liver plasma membranes. Endogenous ligands were removed by prior dialysis of the membranes. Specific binding of the ligands tested was characterized by rapid forward and reverse kinetics and heterogeneity as indicated by curvilinear Scatchard plots. The KD in the high affinity range was 80 nM for [3H]adenosine, 84 nM for [3H]NECA, and 168 nM for [3H]CHA; the respective binding capacities of 1.19, 1.03, and 1.05 pmol/mg protein were of virtually the same magnitude, suggesting labeling of identical sites. However, all ligands also displayed binding to large numbers of low affinity sites. This high level of apparently non receptor binding markedly influenced the adenosine structure-activity profile of [3H]CHA displacement, which differs with pharmacological findings. - NECA and CHA stimulated hepatic adenylate cyclase with an apparent ED50 of 60 and 580 nM, respectively; adenosine was stimulatory at a concentration range from 0.1 - 2.0 microM, but inhibitory at higher concentrations. Hence, estimation of the true ED50 was not possible. Because the KD of high affinity binding and the ED50 of the biological effect of NECA and CHA are in the same range, it may be reasonable to assume that the high affinity sites represent adenosine receptors, recently classified as Ra-site receptors. PMID- 6287299 TI - Role of globus pallidus GABA and opiate receptors in apomorphine circling in nigro-striatal lesioned rats. AB - The effect of lesioning the ipsilateral globus pallidus (GP) on apomorphine induced circling in nigro-striatal lesioned rats was investigated. A GP electrolesion almost abolished circling whereas a kainic acid lesion partly antagonized circling. Drugs that affect GABA and opiate receptors were injected in GP through a cannula. Circling was antagonized by the GABA antagonists picrotoxin and bicuculline, the GABA agonist muscimol and by baclofen. Opiate receptor agonists including morphine, levorphanol, [D-Ala2, D-Leu5]-enkephalin and beta-endorphin had no effect on circling in GP. Ethylketazocine caused a pronounced, naloxone-reversible slowing of apomorphine circling. Apomorphine induced circling behaviour may be modulated by GP GABA receptors and kappa-type opiate receptors. PMID- 6287300 TI - Comparative pharmacology of mianserin, its main metabolites and 6-azamianserin. PMID- 6287301 TI - Benzamide action at alpha 2-adrenoceptors modifies catecholamine-induced contraction and relaxation of circular smooth muscle from guinea-pig stomach. AB - Dopamine was shown to act on the circular smooth muscle of the stomach body to cause contraction at a yohimbine-sensitive site (alpha 2) and a relaxation at a prazosin-sensitive site (alpha 1). Metoclopramide and tiapride failed to modify either response, failed to antagonise a relaxation to phenylephrine at alpha 1 sites in the same tissue, and failed to modify the contractions caused by dopamine and phenylephrine at an alpha 1-adrenoceptor site in the pyloric sphincter. However, (+)- and (-)-sultopride and (+)-sulpiride antagonised the dopamine-induced contractions of the stomach body indicating an alpha 2 antagonist action. An ability to attenuate the relaxation of this tissue may reflect a displacement of the contraction curve to the right rather than an alpha 1-antagonist action since the response to phenylephrine was not antagonised either in this tissue or in the pyloric sphincter. Within the central nervous system the (-)-enantiomers of sultopride and sulpiride have a highly selective dopamine receptor blocking action. This contrasts with the present findings in the stomach musculature of a non-stereospecific antagonism at alpha 2-type adrenoceptors. PMID- 6287302 TI - Non-cholinergic, non-adrenergic responses to nerve stimulation of different regions of the guinea-pig small intestine. AB - Non-cholinergic, non-adrenergic responses to nerve stimulation recorded from smooth muscles of the guinea-pig duodenum, jejunum, proximal and terminal ileum were investigated in an attempt to characterize these responses. In the presence of atropine (1-2 mumol X l-1) and guanethidine (10 mumol X l-1) coaxial stimulation induced in all regions of the guinea-pig small intestine an initial relaxation (primary relaxation) upon which contraction (primary contraction) appeared, followed by rebound contraction. Noradrenaline decreased the cholinergic smooth muscle twitches, predominantly at low stimulation frequencies, and had a similar effect on the non-cholinergic, non-adrenergic primary relaxation, primary and rebound contractions. ATP decreased the smooth muscle twitches; however, this agent had only a transient influence on the non cholinergic, non-adrenergic responses of muscle (tension and membrane potential) to single stimuli. With higher stimulus frequencies ATP increased the primary relaxation and decreased the contraction phases. ATP also inhibited the post tetanic inhibition induced by non-cholinergic, non-adrenergic nerve stimulation. In most of the muscle cells of the guinea-pig proximal and terminal ileum the non cholinergic, non-adrenergic nerve stimulation generated i.j.p.s., while about 15 20% of the cells responded with e.j.p.s. During long-lasting stimulation (10s) the i.j.p.s were sometimes "interrupted" by action potentials or by a gradual depolarization of the membrane. The i.j.p.s were followed by a marked rebound depolarization accompanying the action potentials. Those cells which generated i.j.p.s in response to field stimulation, were depolarized by ATP, while those cells, which generated e.j.p.s, were hyperpolarized by ATP. A reduction in the concentration of extracellular sodium chloride decreased both the primary and rebound contractions; the primary contraction was, however, more sensitive than was the rebound contraction. Theophylline increased the primary and rebound contractions with no marked influence on the primary relaxation, lowered the action potential threshold, increased the rebound depolarization and did not markedly influence the i.j.p.s. Quinidine enhanced the primary relaxation and inhibited the primary contraction in a concentration-dependent manner. Inhibition of the rebound contraction by quinidine was slight (less than 50%). The present results demonstrate that primary relaxation, primary and rebound contractions are associated with i.j.p.s and e.j.p.s, and rebound depolarization with action potentials, respectively; they are typical responses of various regions of the guinea-pig small intestine to activation of inhibitory and excitatory non cholinergic, non-adrenergic nerves. The P1 and P2 receptors, proposed by Burnstock (1975), probably do not mediate the non-cholinergic, nonadrenergic postsynaptic responses of the guinea-pig small intestine... PMID- 6287303 TI - Role of Ca2+ in non-cholinergic, non-adrenergic responses to nerve stimulation of the guinea-pig small intestine. AB - The effects of calcium channel blockers (D-600, verapamil), sodium nitroprusside, papaverine, indomethacin, local anaesthetics and blockade of sodium pump activity on the non-cholinergic, non-adrenergic transmission in the guinea-pig duodenum, jejunum, proximal and terminal ileum were analysed in the presence of atropine and guanethidine. A decrease of the extracellular Ca2+ concentration inhibited the primary and rebound contractions but only in Ca2+ -free solution was the primary relaxation diminished. D-600, verapamil, sodium nitroprusside and papaverine inhibited both the primary and rebound contractions to the same degree and their effects on the primary relaxation were less pronounced than on the contractions. Indomethacin dissolved in alkaline solution did not depress the non cholinergic, non-adrenergic responses in any region of the small intestine, whereas indomethacin dissolved in ethanol antagonized both the primary and rebound contractions in the muscles. Local anaesthetics (procaine, trimecaine) in low concentrations inhibited only the primary contraction. Higher concentrations also inhibited both the rebound contraction and primary relaxation. Procaine in low concentrations did not markedly affect the non-cholinergic, non-adrenergic i.j.p.s and e.j.p.s, but did block the action potentials induced by e.j.p.s. Our findings indicate that the primary relaxation, and primary and rebound contractions are probably induced by different mechanisms and are not mediated by ATP. We confirmed that prostaglandins did not participate in the generation of the rebound contraction. PMID- 6287304 TI - Opiate receptor binding studies in the mouse vas deferens exhibiting tolerance without dependence. AB - The apparent lack of dependence in highly opiate-tolerant isolated mouse vas deferens may conflict with unitary theories postulating a common biochemical mechanism for both phenomena. Therefore, attention focused on binding sites of the opiate receptors which in this tissue are located presynaptically. Binding studies conducted with homogenate of highly tolerant vasa deferentia revealed no significant different results as compared to naive tissues. Further studies examined the effect of guanine nucleotide on opiate receptor interaction. Apparently, mu-, delta- and kappa-opiate receptors in the mouse vas deferens proved resistant to the regulatory action of guanine nucleotide in naive and tolerant tissues. From these experiments, it is concluded that the binding characteristics of opiate receptors in the mouse vas deferens do not change with chronic activation. In addition, the lack of an effect of guanine nucleotide on opiate binding leads to the suggestion that binding sites are not coupled to adenylate cyclase in this tissue. Taken together, these findings draw the attention to the coupling mechanism of opiate receptors in the mouse vas deferens which may play a key role in the adaptational mechanisms following chronic opiate exposure. PMID- 6287305 TI - Enhancement of delta- but not mu-opiate agonist binding by calcium. AB - Present evidence for distinction of 2 types of opiate receptor sites in rat brain homogenates originates from different relative affinities of morphine-like alkaloids and enkephalins to delta- or enkephalin and mu- or morphine-receptor sites. We now report that Ca2+ in a physiological dose range (0.5-3 mM) enhances the binding of 3H-enkephalin in hypotonically treated rat brain membranes, whereas specific binding of 3H-morphine-like alkaloids is not affected. Furthermore, the potency of [D-Ala2,D-Leu5]-enkephalin to inhibit [3H] diprenorphine and [3H]-ethylketazocine binding increased in the presence of Ca2+, whereas an increase in potency of [D-Ala2,D-Leu5]-enkephalin to inhibit binding of mu-receptor ligands was not observed. Kinetic analysis revealed that Ca2+ decreased the rate of dissociation of [D-Ala2,D-Leu5]-enkephalin without affecting the rate of association, thereby increasing the affinity. However, in saturation binding studies, performed in diencephalic membranes, in which [D Ala2,D-Leu5]-enkephalin binds predominantly to mu-receptors, Ca2+ also increased the binding affinity of [3H]- [D-Ala2,D-Leu5]-enkephalin. Double reciprocal analysis suggested a mixed competitive-noncompetitive type of inhibition of [D Ala2,D-Leu5]-enkephalin binding by dihydromorphine. Thus, the interaction of delta- and mu-opiate ligands with mu-receptors may involve topographically different, but closely related binding sites, located on a single receptor molecule. PMID- 6287306 TI - Evidence for a neuromodulatory role of GABA at the first synapse of the baroreceptor reflex pathway. Effects of GABA derivatives injected into the NTS. AB - Microinjections of GABA and of the specific agonist of GABA receptors, muscimol, in the intermediate nucleus tractus solitarii (NTS) of pentobarbitone anaesthetized cats produced hypertension and tachycardia. The GABA receptor antagonist, bicuculline, had opposite effects and prevented those of muscimol. Therefore, a GABAergic system appears to modulate the cardiovascular regulation within the NTS. d,l-Baclofen also increased blood pressure and heart rate when injected into the same region, but this effect was not antagonized by bicuculline. The mechanism of this action of baclofen is discussed. PMID- 6287307 TI - Beta-carboline binding indicates the presence of benzodiazepine receptor subclasses in the bovine central nervous system. AB - Receptor binding studies were performed with tritiated propyl beta-carboline-3 carboxylate ( [3H]PrCC), tritiated ethyl beta-carboline-3-carboxylate ( [3H]ECC), and tritiated flunitrazepam ( [3H]FNT) in membrane preparations from different regions of the bovine brain and retina. Specific binding in all regions investigated was associated with benzodiazepine receptor sites. However, not all benzodiazepine receptors in the regions investigated as determined by the specific binding of tritiated flunitrazepam ( [3H]FNT) are available for [3H]PrCC suggesting that specific [3H]PrCC binding labels only one subclass or subpopulation of the benzodiazepine receptor. This benzodiazepine receptor subclass is sensitive to GABAergic modulation and amounts for about 60% of the benzodiazepine receptors in bovine cortex, hippocampus, and retina but for about 80% of the benzodiazepine receptors in the bovine cerebellum. By contrast, specific [3H]ECC binding in the cerebellum and the hippocampus labeled the same number of benzodiazepine receptors as [3H]FNT, giving no evidence for a benzodiazepine receptor subclass specificity of this compound in the bovine CNS. PMID- 6287308 TI - The effect of opiates on arterial baroreceptor reflex function in the rabbit. AB - The contribution of exogenous and endogenous opioid peptides to the central modulation of the baroreceptor reflex was investigated in rabbits. Baroreceptor sensitivity was assessed in pentobarbitone anaesthetised animals by measuring heart period in response to rises in arterial pressure after bolus intravenous injections of phenylephrine and falls induced by intravenous sodium nitroprusside and controlled haemorrhage. The slope of the linear relationship between arterial pressure and heart period was used as an index of baroreflex sensitivity. Thirty minutes after the intracisternal administration of 50 microgram/kg RX783016 (a mu opiate receptor agonist) baroreceptor sensitivity was reduced to all three methods of blood pressure manipulation. Ketazocine (50 microgram/kg) a kappa opiate agonist and [D-Ala2, D-Leu5] enkephalin (1 microgram/kg) a delta-opiate agonist, 15 min after intracisternal injection caused an increase in baroreflex sensitivity in response to a rise in pressure and a reduction in response to a fall. Intravenous injection of naloxone (80 microgram/kg) caused an increase in varoreflex gain. However, a higher dose (200 microgram/kg) was required to attenuate the effects of RX783016 and [D-Ala2, D-Leu5] enkephalin but not ketazocine. No change in baseline arterial pressure or heart rate occurred after the opiates or naloxone. It appears that exogenous and endogenous opiates modify baroreceptor reflex function, through a mechanism which involves central opiate receptors of the mu- and kappa-types. PMID- 6287309 TI - An initial characterization of human heart beta-adrenoceptors and their mediation of the positive inotropic effects of catecholamines. AB - The positive inotropic effects of catecholamines were studied on samples of ventricular myocardium taken from patients undergoing open heart surgery. The adenylyl cyclase and binding of 3H-(-)-bupranolol were examined in membrane particles prepared from similarly obtained samples. The equilibrium dissociation constant (KD) for (-)-bupranolol was estimated in 4 ways: blockade of the positive inotropic effects of catecholamines, blockade of the stimulation of the adenylyl cyclase by catecholamines, saturation binding of 3H-(-)-bupranolol, inhibition of the binding of 3H-(-)-bupranolol by its unlabeled stereoisomers. The estimates of KD fall in the range 0.5-1.4 nmol/l. The stereo-selectivity ratio (KD (+)-isomer/KD (-)-isomer) is 73. Both values for bupranolol are very similar in cat and man. The inotropic potency of (-)-noradrenaline is nearly 2 orders of magnitude higher in cat heart tissues than in tissues from human hearts. The difference in inotropic potencies between species is only partially accounted for by the five-fold lower potency of (-)-noradrenaline for the human heart adenylyl cyclase as compared to the cat enzyme. PMID- 6287310 TI - Effect of diamide and reduced glutathione on the elevated levels of cyclic AMP in rat pancreatic islets exposed to glucose, p-chloromercuribenzoate and aminophylline. AB - In rat pancreatic islets the effects of diamide, which has been shown to decrease islet levels of reduced glutathione (GSH), and of exogenous GSH were investigated on cyclic AMP as increased by glucose, p-chloromercuribenzoate, and aminophylline. In addition the effect of diamide on islet ATP level, low Km and high Km phosphodiesterases was studied. Diamide (0.1 mM) inhibited the increase of cyclic AMP (cAMP) in response to glucose (16.7 mM), and p-chloro mercuribenzoate (1 mM) in the presence of 5.6 mM glucose. No inhibitory effect of diamide could be demonstrated when cAMP was raised by 10 mM aminophylline in the presence of 5.6 mM glucose. The glucose (27.7 mM) stimulated increase of cAMP was further augmented by GSH (0.4 mM) whereas GSH in the presence of 5.6 mM glucose had no such effect. Diamide neither affected islet high Km nor low Km cAMP phosphodiesterases. Diamide (0.1 mM) as used in this study did not affect islet AMP levels. A concentration dependent decrease of ATP was observed, however, with higher concentrations of diamide (0.25, 0.5 and 1.0 mM). It is suggested that the accumulation of islet cAMP in response to glucose and para-chloromercuribenzoate depends on the redox state of islet thiols. Since thiol oxidant diamide neither affected cAMP-phosphodiesterase activities nor inhibited aminophylline induced accumulation of cAMP in the presence of low glucose the possibility is raised that in pancreatic islets the formation of cAMP rather than its degradation depends on the redox state of islet thiols. PMID- 6287312 TI - [After care and follow up; doctor versus patient]. PMID- 6287311 TI - Chemical modification of membrane proteins by brominated taurodehydrocholate in isolated hepatocytes; relationship to the uptake of cholate and of phalloidin and to the sensitivity of hepatocytes to phalloidin. AB - In vitro treatment of isolated rat hepatocytes with brominated taurodehydrocholic acid (BTC) reduced their sensitivity against phalloidin and inhibited the uptake of phalloidin as well as of cholate in an irreversible and concentration dependent manner. BTC was taken up itself by liver cells; this process was inhibited by 4,4'-diisothiocyano 2,2'-stilbene disulfonate (DIDS). When hepatocytes were incubated with 35S-BTC their plasma membranes contained five labeled protein species with molecular weights of 67,000, 49,000, 38,000, 32,000 and 24,000 as shown by SDS-electrophoresis. No marked difference was observed when isolated plasma membranes from livers were directly treated with the affinity label. DIDS suppressed covalent binding of 35S-BTC to membrane components drastically. Incubation of phalloidin insensitive AS-30D ascites hepatoma cells with 35S-BTC did not result in a chemical modification of the above five proteins. This agrees with an earlier observation that hepatoma cells are unable to take up phalloidin and bile acids (Petzinger et al. 1979; Rufeger and Grundmann 1977; Kroker et al. 1978). PMID- 6287313 TI - [Effect of noradrenaline and serotonin on synaptic transmission in the rat spinal cord]. PMID- 6287314 TI - [Evaluation of the interrelations among auditory potential parameters by means of the technic of correlation-regression analysis]. AB - The characteristics (amplitude, latency) of two types of evoked potentials, action potential of the 8th nerve recorded at the cochlea round window and sonomotor evoked potential recorded from the postauricular region and the back of the guinea pigs were studied in response to sound stimulation. The degree and type of interrelation between the parameters of the auditory evoked potentials were established employing the method of correlation-regression analysis. A close linear dependence was established for amplitude values ( r = 0.93) of the action potential and the sonomotor potential irrespective of the recording site of the latter. The similar dependence (r = 0.91) was established when analysing the latency of the action potential and the evoked sonomotor potential, if the latter was in the postauricular region. The performed mathematical simulation permitted justifying the correction of disturbed functions of the auditory system. PMID- 6287315 TI - [Neurophysiologic analysis of the development of cortico-striatal connections during postnatal ontogeny in the rabbit]. AB - Responses of the caudate nucleus to electrical stimulation of the ipsilateral motor cortex were recorded in 2-30 day old and adult rabbits. The caudate response could be evoked already in the youngest (3 day old) animals. The configuration and topography of these evoked potentials were identical to the responses in adult rabbits. The latency was about 10 ms in the youngest rabbits. A significant decrease of the latency occurred between the 16-17th and 20th day. The definite values of amplitude and latency were reached to the 30th day. The results suggest an early maturation of the corticostriatal input in rabbit and correlate with its early structural maturation. PMID- 6287316 TI - [Effect of intracellular injection of cyclic adenosine monophosphate on the calcium current in identified snail neurons]. AB - The effect of intracellular cyclic adenosine monophosphate (cAMP) injection and extracellular theophylline application on calcium current were investigated in Helix RPa3 and LPa3 neurons. It was found that iontophoretic cAMP injection (current 10-35 nA, duration about 1 min) caused a decrease in the amplitude of calcium current which restored to the initial level following the cessation of injection. Its current-voltage characteristic was not displaced along the potential axis in the presence of cAMP injection indicating that the calcium current decrease was due to the fall of maximum calcium conductance. Extracellular theophylline application in concentration of 1 mM/1 caused a decrease in the calcium current amplitude by 50-75% from the initial level. The hypothesis is discussed about the participation of cytoplasmic factors in regulation of calcium current in mollusc neurons. PMID- 6287317 TI - [Blockade of the sodium and potassium channels of a myelinated nerve fiber by quinidine]. AB - The effects of quinidine on sodium (INa) and potassium (IK) currents in the Ranvier node of frog myelinated nerve fibre was studied by means of voltage clamp technique. When applied externally quinidine (5.10(-5) M) suppresses both INa and IK. Inhibition of INa can be greatly increased by repetitive membrane depolarization. After the end of stimulation the INa value recovers slowly up to the initial level (time constant being about 30 s at 12 degrees C). Unlike repetitive stimulation a single depolarizing pulse of long (1s) duration does not enhance appreciably the quinidine block, which permits a conclusion that quinidine interacts preferently with open sodium channels. Batrachotoxin protects the channels from the blocking action of 5.10(-5) M quinidine. The outward IK is blocked by quinidine in time- and voltage-dependent manner suggesting the interaction of the drug with open potassium channels. The results are consistent with the notion that tertiary amine quinidine, like amine local anesthetics penetrates through the membrane in the neutral form and blocks open sodium and potassium channels from inside in charged (protonated) form. Quinidine and local anesthetics are supposed to share a common receptor in the inner mouth of the sodium channel. PMID- 6287318 TI - Electrochemical properties of polycyclic compounds studied by the polarographic method in anhydrous systems. VIII. The influence of proton-donor on reduction of carcinogenic nitrogen compounds in dimethylformamide. AB - The authors have studied the electrochemical behavior of a series of carcinogenic and inactive heterocyclic compounds in anhydrous medium in the presence of a proton-donor. As anhydrous medium the authors used anhydrous dimethylformamide as proton-donor phenol. In the presence of proton-donor there had occurred on polarographic reduction of the employed carcinogenic heterocyclic compounds changes in the number of polarographic waves. Between the two original waves in these compounds there arose in the presence of phenol a new wave. In non carcinogenic heterocyclic compounds such an effect of phenol was not noted. In the present paper the authors discuss the possible mechanism of electroreduction of carcinogenic and inactive heterocyclic compounds in anhydrous medium in the presence of a proton-donor. PMID- 6287319 TI - Characterization of murine leukemia viruses activated during N-nitroso-N methylurea-induced leukemogenesis in mice. I. Host range of activated viruses. PMID- 6287320 TI - Reduced collagenolytic activity of rat kidneys with steptozotocin diabetes. AB - Collagenolytic activity of rat kidneys with streptozotocin diabetes was estimated by means of a biological collagenase assay and compared to healthy controls. Collagenolytic activity was found significantly decreased in rat kidneys with diabetes correlating with blood glucose levels (r = -0.82, p less than 0.001). Elevated blood glucose levels seem to be responsible for the inhibition. This is supported by our experiment of incubating bacterial collagenase with several carbohydrates as glucose, galactose and saccharose: glucose and galactose significantly inhibited the collagenolytic activity, while saccharose failed to inhibit the enzymatic reaction. The interpretation of the results is that glucose is able to bind to the enzyme as Schiff base, which could be shown by tritiated sodium borohydride reduction of the Schiff base formed between collagenase and glucose. Another support of the hypothesis is that blocking of the amino group of lysine at the active site either by glucose or trifluoroacetylation of collagenase is reducing the collagenolytic activity. The biological significance could be the decreased catabolism of collageneous material of the extracellular matrix, as, e.g., the glomerular basement membrane, which was reported in a previous publication. PMID- 6287321 TI - ["Rehabilitation" in malignant brain tumours? (author's transl)]. PMID- 6287322 TI - [Pituitary adenoma and meningioma in the same patient (author's transl)]. PMID- 6287323 TI - Meningioma as a sequel of radiotherapy for pituitary adenoma. AB - Two cases of meningioma developing after radiation therapy of pituitary adenoma were found among 180 verified adenomas irradiated pre- or post-operatively and followed up for at least five years. Only a few instances of association of meningioma and pituitary adenoma were found in the literature. In patients who did not receive X-ray treatment both tumours were simultaneously verified at operation and/or autopsy, whereas in those who were irradiated a definite time interval, ranging from 11 to 18 years, was present between surgical and/or radiotherapeutic treatment for pituitary adenoma and verification of meningioma. A possible role of radiation in the genesis of these meningiomas is postulated. PMID- 6287324 TI - [Fracture dislocation of the sacral spine with cauda equina lesion. Report of two cases (author's transl)]. AB - Transverse sacral fractures with cauda equina lesion appear only sporadically in the world literature. In our medical center 667 spine fractures were admitted during a twelve year period and we report only 2 cases of sacral fractures with neurological lesions (0,20%). The neurological abnormalities that occur are a perianal sensory loss with bladder and anal sphincters disturbance. Sphincters electromyographic studies and cystometric examination show a lower motor neuron lesion. In one of our cases, the nervous lesion was more spread with an unilateral sensory loss S1-S2 and a motor loss L4-L5 S1 in the same side. The high level of sacral lesion (S1-S2), the association with other pelvic fractures and fractures of the lower lumbar transverse processes, suggests the mechanism of injury (sudden flexion). Radiological studies show that sacral fractures are often difficult to detect (necessity of lateral view). Lumbar metrizamide exploration seems not very interesting. Sacral laminectomy which permits the decompression of sacral roots appears like the best treatment. The post-operative course, studied by clinical findings, electromyographic and cystometric examinations, shows the habitual resolution of neurological deficit (incomplete nevertheless in our 2 patients). PMID- 6287325 TI - Microtubular organization in flat epitheloid and stellate process-bearing astrocytes in culture. AB - Microtubules and microfilament patterns in cultured astrocytes were revealed by using indirect immunofluorescent microscopy in conjunction with anti-tubulin immune serum and anti-actin immunoglobulins respectively. In flat epitheloid astroglial cells (either polygonal or elongated) colchicine-sensitive immunofluorescent fibres, which correspond to bundles of microtubules, extend from the perinuclear cytoplasm into the cell periphery by running for long distances through the different focal planes. These patterns of organization differ markedly from the patterns of organization of microfilaments which are arranged in fibres parallel to each other and often oriented along the cell boundary. In response to the combined treatments of serum withdrawal and administration of dBcAMP, flat epitheloid astrocytes adopt a morphology similar to that of the mature astrocytes in situ in the CNS, that is of stellate process bearing cells. This is prevented or is reverted by the administration of colchicine at the appropriate times. There are strong suggestions indicating that during cell processes formation the microtubular network is reorganized and microtubules assembled into dense bundles which are oriented along the axis of the cell processes. In view of these results, we suggest that, in contrast to microfilaments, microtubules are not determinant for the maintenance of cellular shape in elongated or polygonal flat epitheloid astroglial cells but they are required for both the formation and maintenance of processes in stellate astrocytes. PMID- 6287326 TI - Differential effects of metal ligands on synaptic membrane glutamate binding and uptake systems. AB - The high affinity, Na+-independent L-[3H]glutamate binding process in synaptic membranes and in the purified binding protein was shown to be inhibited to an almost equal extent by the metal ligands NaN3, KCN, and o-phenanthroline, and by 2,4,5-trihydroxyphenylalanine (6-OH DOPA). The high affinity, Na+-dependent glutamate transport activity in these membranes was almost totally insensitive to NaN3, o-phenanthroline, KCN, and 6-OH DOPA. These agents, especially 6-OH DOPA, may be useful tools in achieving a discrimination between putative physiologic receptors and uptake carrier sites for L-glutamate in synaptic membranes. The sensitivity of the glutamate binding sites to the effects of the metal ligands may be correlated to the presence of an iron-sulfur center in the purified glutamate binding protein. some of the characteristics of this metallic center were explored by optical and paramagnetic resonance spectroscopic techniques and are described in this study. PMID- 6287327 TI - [3H]GABA binding in developing rabbit retina. AB - We have studied the developmental sequence of the GABA system in the rabbit retina using an in vitro binding assay to monitor developmental changes in the post-synaptic receptor. A variety of tissue treatments including perchlorate and Triton X-100 were employed to optimize binding and remove endogenous factors which inhibit binding. Pre-treatment of the tissue with 0.05% Triton X-100 revealed high affinity binding for [3H]GABA which increased in a sigmoidal fashion with the post-natal age of the animal. A constant level of binding, at about 16% of adult levels, was noted until day 8, at which time a rapid increase occurred. At 16 days post-natal, the amount of specific binding reached a plateau near adult levels. Kinetic analysis of the GABA receptor showed an increase in the number of receptors (Bmax) with little or no change in the apparent affinity (KD). Our results suggest that the onset of post-synaptic receptor activity is delayed approximately 1 to 2 days, relative to the pre-synaptic components, and the period of rapid increase in GABA receptor binding coincides with the period of maximum increase in retinal synaptic density. PMID- 6287328 TI - Benzodiazepine antagonist Ro 15-1788 counteracts the prolactin-lowering effects of other benzodiazepines in rats. AB - A number of centrally active benzodiazepines lowered baseline serum prolactin concentrations after oral administration to male rats. The increase of circulating prolactin levels elicited by oral administration of various neuroleptic agents was also reduced by prior or simultaneous oral administration of several benzodiazepines in a dose-dependent manner. Since both effects were prevented by simultaneous administration of the benzodiazepine antagonist Ro 15 1788 they are probably mediated by central benzodiazepine receptors which interfere with aminergic mechanisms governing serum prolactin. PMID- 6287329 TI - Resumption with clonidine of pulsatile LH release following acute norepinephrine depletion in ovariectomized rats. AB - The effects of adrenergic stimulation on the pulsatile release of LH were investigated in ovariectomized rats with acute depletion of brain norepinephrine (NE) levels. Rats bearing atrial cannula were pretreated with NE synthesis inhibitors, diethyldithiocarbamate (DDC) or bis (4-methyl-1-homopiperanzinyl thiocarbanyl) disulfide (FLA-63) and blood was withdrawn at 15-min intervals beginning 1 h later. DDC and FLA-63 markedly dampened pulsatile LH secretion. Administration of the alpha-adrenergic agonist clonidine (CLON) resulted in immediate LH release and apparent resumption of pulsatile LH secretion. The facilitatory effect of CLON on LH secretion was was more pronounced in FLA-63 than in DDC-pretreated rats. Additional characterization of the pattern of LH secretion in FLA-63-pretreated rats showed that the interval between LH pulses was significantly lengthened after acute NE depletion; however, CLON treatment increased LH pulse frequency to that found in ovariectomized rats. To further investigate the ability of CLON to augment pulsatile LH release, the LH secretory pattern was determined between 2 and 4 h after CLON administration FLA-63 pretreated rats. 8 of 10 rats receiving CLON (0.3 mg/kg) responded with episodic LH release 2-4 h following treatment while saline-treated rats continued to show dampened LH secretory patterns. These studies demonstrate that following acute depletion of NE in ovariectomized rats, a single injection of CLON can enhance both the amplitude and frequency of LH pulses. Further, the data suggest that central noradrenergic neurons exert only a permissive effect on pulsatile LH release and that the pulsatile mechanism may predominately be resident in LHRH neurons. PMID- 6287330 TI - Differentiation of excitatory amino acid receptors in the rat caudal trigeminal nucleus: a microiontophoretic study. AB - The excitatory amino acid antagonist, D-alpha-aminoadipate, was found to be a potent antagonist of responses to N-methyl-D-aspartate and L-aspartate, but less potent against L-glutamate responses of neurones in the rat caudal trigeminal nucleus. The responses to quisqualate and kainate were relatively unaffected by D alpha-aminoadipate, but were antagonized by the two antagonists cis-2,3 piperidine dicarboxylate and gamma-D-glutamylglycine. These two antagonists had effectively similar spectra of activity. It is concluded that there is a distinct N-methyl-D-aspartate receptor in the trigeminal nucleus caudalis, and that kainate and quisqualate may act on a different, but common, receptor. Exogenously applied L-glutamate appears to act at both of these receptors, but exogenously applied L-aspartate appears to act almost exclusively on a D-alpha-aminoadipate sensitive site that may be the N-methyl-D-aspartate receptor. The kainate/quisqualate receptor is thought to be involved in the synaptic excitation of neurones in the caudal trigeminal nucleus by non-noxious mechanical stimuli. PMID- 6287331 TI - Evidence that SL75102 is an agonist at GABAb as well as GABAa receptors. AB - The compound SL75102 ([alpha(4-chlorophenyl)5-fluoro 2-hydroxy benzilidene-amino] 4-butanoate sodium) is a GABA-mimetic at bicuculline-sensitive and -insensitive receptors. It depolarized rat isolated superior cervical ganglia in a dose dependent manner (relative potency = 0.101 +/- 0.013; GABA = 1). Bicuculline methobromide (13 microM) antagonized this action of SL75102 and shifted the log dose-response curve to the same extent as the GABA curve. The evoked release of [3H]noradrenaline from rat isolated atria was also reduced by SL75102 in a GABA like manner. SL75102 also displaced [3H]GABA and [3H]baclofen specifically bound to divalent cation dependent GABAB sites on rat synaptic membranes (relative potency approximately 0.1 GABA throughout). The butyramide from which SL75102 can be formed within the body (SL76002) was much less active at GABAB sites (less than 0.02 atria, 0.001 binding, GABA = 1). It is suggested that in addition to any direct action of SL76002 itself the products of SL76002 metabolism, SL75102 and GABA may exert effects via baclofen-sensitive GABAB as well as GABAA sites in mammalian brain. PMID- 6287332 TI - A micromethod for the measurement of renin in brain nuclei: its application in spontaneously hypertensive rats. AB - The aim of this study was to develop a method for the measurement of renin activity in small tissue samples obtained from rat brains by the micropunch technique and to investigate the activity of brain renin in spontaneously hypertensive rats. The assay satisfied sensitive and specificity requirements. Angiotensin I was generated at a pH of 6.0; complete recovery of angiotensin I and kinetic studies supported the specificity of the method. Angiotensinase and cathepsin D-like acid protease activity were measured in parallel with renin. Renin was present in all brain regions studied and decreased with the age of the animals. An increased activity of renin was measured in several nuclei of the brain stem and in the neurohypophysis of young hypertensive rats when compared with age-matched normotensive control animals. These differences disappeared in older rats. There was a dissociation between renin and cathepsin D-like acid protease activity. No correlation existed between the distribution of renin and angiotensinase activity. The increased renin activity in brain stem nuclei of spontaneously hypertensive animals is in agreement with previous findings that the brain renin-angiotensin system contributes to the maintenance of high blood pressure in these rats. PMID- 6287333 TI - Comparison of the properties of central and peripheral binding sites for cocaine. AB - Cocaine and its analogs bound saturably to membranes of brain and liver of mice. The binding sites on membranes of liver had a lower affinity for cocaine than those of brain. In addition, there were striking differences between the two tissues in regard to the relative potencies of cocaine analogs in competing with [3H]cocaine for binding. In comparison with the binding sites in brain, those in liver had only moderate stereospecificity, and they discriminated less between the centrally active compounds and the centrally inert analogs. PMID- 6287334 TI - Multiple opiate receptors: support for unique mu, delta and kappa sites. PMID- 6287335 TI - Responses of cortical neurones to stimulation of the nucleus raphe medianus: a pharmacological analysis of the role of indoleamines. AB - Single shock stimulation of the nucleus raphe medianus evoked complex responses from the large majority of neurones tested. These responses consisted of a short latency (mean 21 msec) inhibition of firing followed by a longer latency (mean 160 msec) increase in firing rate. Occasionally, cells were encountered which exhibited pure inhibitory or pure excitatory responses. Prior treatment with the tryptophan hydroxylase inhibitor rho-chlorophenylalanine reduced the excitatory effects of raphe stimulation but greatly increased the inhibitory effects. Pretreatment with 5,7-dihydroxytryptamine (5,7-DHT), on the other hand, reduced both inhibitory and excitatory effects of raphe stimulation. Methysergide was found to be an effective antagonist of excitatory responses to iontophoretically applied 5-hydroxytryptamine (5-HT) but less effective against depressant responses to either 5-HT or tryptamine. In contrast metergoline consistently antagonized 5-HT and tryptamine-induced depressions but not 5-HT elicited excitations. When tested against stimulation evoked responses of cortical neurones methysergide antagonized the excitatory effects of raphe stimulation but had much less effect on the inhibitory responses, while the reverse held true for metergoline. The present results may be compatible with a mediation of the excitatory effects of stimulation by 5-HT and the inhibitory effects by tryptamine. PMID- 6287336 TI - Central cardiovascular effects produced by the GABA receptor agonist drug, THIP. AB - In chloralose-anesthetized cats, injection of THIP (30-1000 microgram) into the fourth ventricle produced dose-dependent decreases in blood pressure and heart rate. Administration of similar doses into either the lateral and third ventricle or administration of the largest dose intravenously did not produce these effects. Microinjection of THIP (1.95 microgram) into nucleus ambiguus reversed bradycardia evoked by microinjection of bicuculline methiodide (160 ng) into this nucleus. These results confirm previous findings indicating that activation of GABA receptors in the hindbrain produces hypotension and bradycardia as well as reversal of bicuculline-induced activation of cardiac parasympathetic nerves. PMID- 6287337 TI - On the configuration of the receptors for excitatory amino acids. AB - Separate receptors are recognized for the excitation of mammalian neurones by (a) L-glutamic and quisqualic acids and (b) N-methyl-D-aspartic (NMDA), and other amino acids which have conformationally restricted molecules. Several other compounds, both agonists and antagonists, have been examined, and it is concluded that (i) the NMDA receptor reacts preferentially with substances in a relatively extended configuration, (ii) the glutamate/quisqualate receptor prefers folded molecules and (iii) the distance separating the amino group from the distal anionic function is the critical one determining receptor preference. PMID- 6287338 TI - Neuronal location of N6-cyclohexyl[3H]adenosine binding sites in rat and guinea pig brain. AB - N6-cyclohexyl[3H]adenosine, ([3H]CHA), was used to label adenosine A1 receptors in crude synaptic membranes prepared from rat and guinea pig brain. The density of [3H]CHA binding sites was highest in the rat hippocampus and cerebellum and in the guinea-pig hippocampus. A micro-dissection of coronal sections of guinea-pig hippocampus revealed that the specific binding capacity for [3H]CHA in area CA-1 was 20-30% higher than in area CA-3, dentate gyrus and subiculum. Selective neuronal lesions of serotonergic, noradrenergic and cholinergic afferents to the hippocampus failed to alter [3H]CHA binding to hippocampal membranes. These results suggest that [3H]CHA binding sites are not associated with axons or terminals of these neurones in the hippocampus. Intrahippocampal injection of kainic acid reduced the number of [3H]CHA recognition sites by 30% with no alteration in the affinity of [3H]CHA for these receptors. Thus, a significant portion of A1 receptors may be associated with intrinsic neurones of the hippocampus which do not appear to be innervated by noradrenergic, cholinergic or serotonergic axons. PMID- 6287339 TI - A study of the actions of bromide on frog sympathetic ganglion. AB - The effects of sodium bromide on the bullfrog sympathetic ganglion were studied by extracellular and intracellular recording techniques. Equimolar replacement of sodium chloride (112 mM) by sodium bromide in Ringer's solution caused hyperpolarization (means = 7.4 mV) of ganglion cells, and antidromically evoked spikes showed increased rates of rise as well as prolonged post-spike positivity. These effects, rapid in onset, returned to control values after approximately 30 min of continued exposure to bromide Ringer. Orthodromic synaptic transmission was not impaired by bromide (84-112 mM); instead it produced increased post-spike negativity and occasionally, stimulus-bound repetitive postganglionic responses (SBR) to each single preganglionic stimulus. This synaptic enhancement persisted as long as exposure to bromide continued. Since ethanol also causes responses, its interaction with bromide was tested and pronounced synergism was found in which 95% of ganglion cells displayed repetitive orthodromic responses to each single preganglionic stimulus. The present and previous studies suggest that bromide and ethanol act at the unmyelinated presynaptic nerve terminal to generate stimulus-bound repetitive responses. The transient nature of the hyperpolarization produced by bromide argues against a mechanistic role in its anticonvulsant action. The persistent synaptic excitatory effects of bromide, however, may have implications for the role of chloride channels in anticonvulsant mechanisms or in epileptogenesis. PMID- 6287340 TI - Effects of chronic treatment with imipramine on the behavioural and electroencephalographic modifications induced by clonidine in the rat. AB - The influence of repeated administration of imipramine on the EEG and behavioural effects of clonidine has been studied in the rat bearing chronic electrodes. Clonidine induced behavioural depression and EEG synchronization in control rats. Mydriasis, hyperirritability, stereotyped behaviour and EEG desynchronization were elicited by clonidine on the first and second days after discontinuation of the treatment with imipramine (15 mg/kg, i.p., daily for 21 days). On the 9th. day the animals responded to clonidine with sedation and EEG synchronization. These results support the hypothesis that chronic treatment with tricyclic antidepressant drugs changes the sensitivity of central noradrenergic neurones. PMID- 6287341 TI - Familial primary nervous system neoplasms in three generations. PMID- 6287342 TI - Regulation of genetic activity of thyroid hormones. I. Effect of purified cytoplasmic hormone-receptor complex on the template activity of chromatin. PMID- 6287343 TI - Localization of ATPase in the cerebral cortex and its role in neuronal functional activity. PMID- 6287344 TI - The dorsal column nuclei of the frog. AB - Surface potentials and field potentials recorded from the medulla in response to cutaneous and mixed nerve stimulation revealed that fibers of the dorsal white column project onto two separate neuron groups in the medulla representing the dorsal column nuclei. The hind-limb was represented in the medial group and the fore-limb in the lateral group in the projection. The fore-limb nerves projected to a region extending from the medulla to the 4th segment of the spinal cord. Primary afferent depolarization and depression of synaptic activities were shown by direct and indirect stimulation of dorsal column fiber terminals in the dorsal column nuclei. Conditioning volleys set up in hind-limb nerves had no effect on test responses evoked by stimulation of forelimb nerves, and vice versa. Slow negative potentials with decreasing latencies were recorded from the posterocentral nucleus of the thalamus in response to stimulation of the 2nd dorsal root, the dorsal column and the dorsal column nuclei, respectively. The physiological results were correlated with histological observations using the cobalt labelling method. It was concluded that the amphibian dorsal column-medial lemniscus system is closely comparable with that found in the mammalian brain. PMID- 6287345 TI - Acetylcholine may be an excitatory transmitter mediating visual excitation of 'transient' cells with the periphery effect in the cat retina: iontophoretic studies in vivo. AB - The effects of iontophoretically-applied acetylcholine and its antagonists on the response of retinal ganglion cells at the receptive field centre were studied in the optically intact eye of anaesthetised cats. Acetylcholine enhanced visually driven excitation of all 'transient' cells with the property of a periphery effect, but failed to excite those weak 'transient' cells without a periphery effect. It also mimicked visual stimulation when applied iontophoretically on spontaneously firing 'transient' cells with a periphery effect and a pulse of acetylcholine produced a transient response. Visually-driven excitation at the receptive field centre and the periphery effect, as well as the acetylcholine induced excitation in these 'transient' cells, were blocked by dihydro-beta erythroidine but not by atropine applied by iontophoresis. Neither acetylcholine nor dihydro-beta-erythroidine, on the other hand, produced any significant effect on the visually-driven firing of 'sustained' cells. Acetylcholine, however, suppressed the spontaneous firing of 'sustained-on' cells with a relatively high rate of background discharge, whereas it enhanced that of 'sustained-off' cells with relatively low rates of background discharge, and such effects were again blocked by dihydro-beta-erythroidine. It is suggested that (1), visual excitation of 'transient' cells with the periphery effect is mediated by a cholinergic system and the receptor is nicotinic rather than muscarinic and (2), the spontaneous firing rate of 'sustained' cells may be regulated also by a cholinergic system. PMID- 6287346 TI - Symmetric sarcoid polyneuropathy: analysis of a sural nerve biopsy. AB - A 20-year-old woman with sarcoidosis had uveitis, subacute symmetric sensorimotor neuropathy, and noncaseous granulomas in biopsies of gastrocnemius muscle and sural nerve. Morphologic studies of the nerve confirmed the electrodiagnostic impression of an acute axonal and demyelinating neuropathy. Of 100 teased myelinated nerve fibers, 15% contained myelin ovoids and 24% demonstrated segmental demyelination. Quantitative analysis showed a reduction in the numerical density of myelinated fibers. By electronmicroscopy, unmyelinated fibers were largely spared. The exact mechanism of nerve fiber damage was not determined, but a local effect of the granuloma seemed likely, because most lesions were found in the endoneurium. PMID- 6287347 TI - Mental neuropathy from systemic cancer. AB - Nineteen patients with mental neuropathy secondary to systemic cancer are described. In nine patients, the numb chin was the presenting symptom of a neoplasm. Nine patients had lymphoreticular malignancies, and the others had a variety of solid tumors. Radiograms of the mandible were abnormal in 5 of 12 patients. The cerebrospinal fluid contained malignant cells in two. Resolution, complete or partial, occurred in 16 of 19 patients receiving radiation or chemotherapy, including 8 who received chemotherapy alone. Sixteen of the 19 patients died within 17 months of the onset of the neuropathy. A nontraumatic mental neuropathy should initiate a search for cancer. PMID- 6287348 TI - Basic and clinical electrophysiology of demyelinating diseases. PMID- 6287349 TI - Opiate, enkephalin, and endorphin analgesia: relations to a single subpopulation of opiate receptors. AB - Differences in the receptor mechanisms of opiate analgesia and respiratory depression have been studied with three novel irreversible opiates. A single injection of the irreversible agonist oxymorphazone produces analgesia in mice, lasting over 24 hours. Conversely, the irreversible antagonist naloxazone dramatically reduces the analgesic effectiveness of a variety of opiate alkaloids and enkephalin analogs for over a day. Despite this marked reduction in analgesia after naloxazone treatment, morphine lethality (LD50) is unchanged in similarly treated mice. Receptor binding studies show that naloxazone irreversibly and selectively blocks a subpopulation of opiate receptors (the mu1 sites) to which all classes of opiates and enkephalins bind with highest affinity, whereas the drug has little to no effect on their lower-affinity sites (mu, and delta). The return of high-affinity receptor (mu1) binding to normal levels corresponds closely to the return of analgesic sensitivity and possibly represents receptor turnover in the central nervous system. These studies suggest that both opiate and opioid peptide analgesia is mediated through a single receptor subpopulation distinct from those involved with respiratory depression, and raise the possibility of specific opiate analgesics without respiratory depression. PMID- 6287350 TI - Orthostatic hypotension in familial amyloid polyneuropathy: treatment with DL threo-3,4-dihydroxyphenylserine. AB - We measured plasma norepinephrine levels in patients with familial amyloid polyneuropathy. Patients with orthostatic hypotension had low basal plasma norepinephrine levels, which did not increase after postural change. On the basis of biochemical findings that suggest depletion of peripheral norepinephrine, DL threo-3,4-dihydroxyphenylserine, an immediate precursor of norepinephrine, was given orally. Six hundred mg of this drug induced substantial and sustained elevation of blood pressure for several hours, and plasma norepinephrine content increased. Daily administration for 4 weeks improved postural dizziness and syncope, and daily activity increased. PMID- 6287351 TI - Lumbosacral plexus neuropathy. AB - We describe 10 cases of lumbosacral plexus neuropathy in which no underlying condition was discovered on initial evaluation or on follow-up examination after an average of 6 years. The patients presented with pain and weakness. Recovery was delayed and often incomplete. When the lower plexus is involved, it may be confused with disk disease manifesting as "sciatica." This syndrome may be a counterpart to the well-described idiopathic brachial plexus neuropathy. PMID- 6287352 TI - Physiologic basis for neuroelectric blocking activity in multiple sclerosis. PMID- 6287354 TI - Posttetanic potentiation in a patient with myasthenia gravis. AB - We studied a patient with clinically typical myasthenia gravis (MG) and high serum titer of antibodies to acetylcholine receptor. Unlike the usual response in MG, there was an increment in the amplitude of the electrical response (308% of control) after 10 seconds of voluntary tetanus. Posttetanic facilitation is usually less than 200% in MG and over 200% in Eaton-Lambert syndrome. However, there have been several other cases of typical MG with increments over 200%. Facilitation of this magnitude has also been seen with curare administration in animals and man. However, in myasthenia, as opposed to curare poisoning, competitive blocking is not thought to exist. PMID- 6287353 TI - Skin involvement in familial amyloidotic polyneuropathy. AB - A 3-mm punch biopsy of clinically normal skin was obtained from the forearm of 11 patients from five kinships with familial amyloid polyneuropathy. Seven of the 11 patients had sensory polyneuropathy in the arms as well as the legs. Abnormalities of the autonomic nervous system (nocturnal diarrhea and sphincter abnormalities) were observed in all patients. Four patients had multiple atrophic scars and poorly healed ulcers on the limb; two had petechiae after gentle stroking of the skin. Histopathology revealed amyloid deposits in all 11 skin biopsies (100%). Amyloid infiltration was noted in blood vessels, sweat glands, dermis, and arrector pili muscles. Intracutaneous neural deposits were not found. Skin biopsies from family members at risk may be an effective method of early diagnosis. PMID- 6287355 TI - 2', 3'-cyclic nucleotide 3'-phosphodiesterase in cerebrospinal fluid. AB - The activity of the myelin-associated enzyme 2',3'-cyclic nucleotide 3' phosphodiesterase (CNP) was assayed in the cerebrospinal fluid (CSF) of 107 neurologic patients by a new and sensitive fluorometric method. The activity of CNP was about 20 nmol per hour per milligram protein or 12 nmol per hour per milliliter CSF. At these extremely low levels, the presence of even a small amount of blood (which has slightly greater activity) significantly elevated CNP values. Patients with radicular syndromes had slightly higher than average CNP activities, but there was no difference in enzyme activities of 47 patients with multiple sclerosis and the general neurologic population. CNP activity was not related to stage of demyelinating illness or intrathecal injection of steroid. CNP-like myelin basic protein may be released into the CSF after destruction of myelin, but our results suggest that the enzyme activity is lost in the process. PMID- 6287356 TI - The spectrum of peripheral neuropathy in myeloma. PMID- 6287357 TI - Brainstem auditory evoked responses in hereditary motor-sensory neuropathy: site of origin of wave II. AB - In three patients with hereditary motor-sensory neuropathy (HMSN), type I, brainstem auditory evoked responses (BAERs) showed prolongation of the latency of all major components. Interpeak latency I-III was prolonged, but the III-V interval was normal, and separation of waves I and II accounted for the prolonged I-III interval. Prolongation of I-II interval in this condition, which affects the peripheral nerves, preservation of wave II after cerebral death, and preservation of only waves I and II in the leukodystrophies suggest that wave II may be generated in the intracranial extramedullary portion of the eighth nerve. PMID- 6287358 TI - Eaton-Lambert syndrome: acetylcholine and choline acetyltransferase in skeletal muscle. AB - In biopsied intercostal muscle from six patients with Eaton-Lambert syndrome, we measured acetylcholine content and release and choline acetyltransferase. Both the spontaneous and the KCl-evoked release of acetylcholine were abnormally low. On the other hand, the acetylcholine content and the level of choline acetyltransferase activity were within the range of values earlier found in healthy human intercostal muscle. These results are consistent with the view that the defect in this syndrome lies not in the synthesis or storage of the transmitter but in the mechanism of release itself. PMID- 6287359 TI - Memory decline in the aged: treatment with lecithin and physostigmine. AB - Normal aged subjects were given lecithin and placebo for 5 weeks each in a double blind crossover study. Supraspan tests of memory and learning failed to show any significant changes as a result of these treatments. Addition of a single IV infusion of physostigmine did not improve performance. These findings neither support nor weaken the "cholinergic hypothesis" of cognitive impairment in aging and dementia, but they imply that simple cholinergic hypofunction is unlikely. PMID- 6287360 TI - Longitudinal screening for herpes simplex virus in obstetric patients. PMID- 6287361 TI - [Cellular and biochemical parameters in the broncho-alveolar lavage fluid of smokers]. PMID- 6287362 TI - Fatigue in cat facial mechanoreceptors. AB - Mechanoreceptor fatigue, the reduction in number of impulses with repeated presentations of the same mechanical stimulus, was studied in facial hairy skin of the cat. We found that the variables that influence the magnitude of fatigue are, in order to importance: stimulus duration, interstimulus interval and stimulus amplitude. We also found that in about 40% of all mechanoreceptors studied, phentolamine, an alpha-noradrenergic antagonist, produced periodic reversal of fatigue and in some cases produced an increase in spontaneous firing. PMID- 6287363 TI - Characterization of corticotropin-like immunoreactive peptides in rat brain using high performance liquid chromatography. AB - The molecular nature of corticotropin (ACTH)-related peptides in rat brain has been studied using high performance liquid chromatography (HPLC) and radioimmunoassay. The major ACTH-immunoreactive species in rat hypothalamic extracts coelutes with corticotropin-like intermediate lobe peptide (CLIP); ACTH18-39) on two HPLC solvent gradients, and has 3-4 times more C-terminal than N-terminal immunoreactivity. N-terminal ACTH-immunoreactivity is composed of a number of peaks on HPLC with less than 10% eluting at the position of ACTH. Hypothalamic C-terminal ACTH immunoreactivity is also heterogeneous and resembles in some respect that seen in the rat neuro-intermediate lobe. Around 90% of the alpha-MSH immunoreactivity in the hypothalamus elutes as a single peak in the position of des[N-acetyl] alpha -MSH (ACTH1-13-NH2). PMID- 6287364 TI - Noise analysis of acetylcholine induced current at neuromuscular junctions of the mouse diaphragm. AB - Miniature end-plate currents (mepcs) and membrane noise elicited by acetylcholine (ACh) iontophoresis were investigated at neuromuscular junctions of the mouse diaphragm. All the experiments were performed at a holding potential of -70 mV at a temperature of 19 degrees C. The equilibrium potential of the ACh response was estimated to be near zero; the mepcs displayed a peak amplitude of 2.46 +/- 0.13 nA (mean +/- S.E.) and relaxed exponentially with a time constant to 1.63 +/- 0.11 msec. Single ACh-activated channels had a conductance of 26.5 +/- 1.5 pS and a mean life time of 1.69 +/0- 0.13 msec. PMID- 6287365 TI - Differential effects of GABA on peripheral and central type benzodiazepine binding sites in brain. AB - The binding of the clinically inactive benzodiazepine [3H]RO-5-4864 to brain membranes was investigated. The peripheral type benzodiazepine binding site was demonstrated in brain with an apparent Kd of 1.6 nM and a Bmax of 20 fmol/mg protein. Monophasic Scatchard plots indicate a homogenous population of a high affinity sites. The major inhibitory transmitter GABA, has no effect on [3H]RO-5 4864 binding to extensively wash brain membranes. The peripheral type benzodiazepine binding site found in brain is therefore not modulated GABA. PMID- 6287366 TI - Effects of caffeine and theophylline on adenosine and benzodiazepine receptors in human brain. AB - The binding of various adenosine receptor ligands and of [3H]diazepam, as well as their inhibition of methylxanthines, have been studied in human brain cerebral cortex membranes. Caffeine and theophylline competitively inhibit binding of [3H]cyclohexyladenosine, [3H]diethylphenylxanthine, [3H]phenylisopropyladenosine and [3H]diazepam. Both caffeine and theophylline are more potent as inhibitors of adenosine receptor ligand binding compared to [3H]diazepam binding. Theophylline was more potent than caffeine in its ability to compete with adenosine receptor ligand binding while the reverse was true for [3H]diazepam binding. The meaning of these results for the mode of action of methylxantine is discussed. PMID- 6287367 TI - Cell type-specificity of epidermal growth factor (EGF) binding in primary cultures of early postnatal mouse cerebellum. AB - Binding of 125I-labeled epidermal growth factor (EGF) to cells of cultured early postnatal mouse cerebellar cells was investigated by autoradiography in conjunction with cell type-specific immunolabeling of neurons, astrocytes and oligodendrocytes. By use of tetanus toxin for recognition of neurons and glial fibrillary acidic (GFA) protein and 04 antigen as markers for astrocytes and oligodendrocytes, respectively, it could be shown that oligodendrocytes do not express receptors for EGF within the limits of sensitivity of the autoradiographic method, and that less than 1% of all small neurons (mostly granule cells) and 50-90% of all GFA protein-sensitive astrocytes show detectable levels of EGF binding. PMID- 6287368 TI - cGMP accumulation by spinal cord neurons during the period of neuromuscular junction formation. AB - A 4--5-fold increase in cGMP levels in spinal cord neurons has been demonstrated in vitro and in vivo upon association with skeletal muscle. The increase on cGMP was temporally coincident with the onset of neuromuscular junction formation and independent of any concomitant change in cAMP. The response was specific for spinal cord neurons cocultured with skeletal muscle; glial cells could not be substituted for spinal cord neurons nor could fibroblasts be substituted for muscle cells. The response was not dependent upon action potential activity, synaptic transmission or muscle contractile activity. Muscle conditioned medium was found to produce the largest increase of cGMP in spinal cord neurons of 23 fold. PMID- 6287369 TI - Sex differences in alpha-adrenergic receptors in the rat brain. AB - [3H]Dihydroergocryptine ([3H]DHE) binding to several brain regions was studied in male and female rats, the latter in two different endocrinological states (estrous and diestrous-1). males showed higher [3H]DHE binding than females in hypothalamus, hippocampus and olfactory bulb, while females showed higher levels than males in spinal cord and visual cortex. In addition, intra-sex (that is, estrous versus diestrous-1) differences were detected in the cervico-thoracic spinal cord and cerebellum. Diestrous females presented higher values in both regions. PMID- 6287370 TI - Multiplicity of opiate receptors in different species. AB - Three populations of opiate binding sites (mu, delta and kappa) could be distinguished by displacement studies with the mu-agonist (D)-Ala2-MePhe4-Glyol5 enkephalin and the kappa-agonist [3H](-)-bremazocine in brain membranes of different species. Whereas brain membranes from pig or guinea-pig and human cortical membranes had a proportion of approximately 35-50% kappa-, 29-40% mu-, and 21-30% delta-binding sites, rat membranes contained much less kappa-sites (12% kappa, 62% mu and 25% delta). PMID- 6287371 TI - Chronic blockade of sciatic nerve transmission by tetrodotoxin does not produce central changes in the dorsal horn of the spinal cord of the rat. AB - Rat sciatic nerves were treated with tetrodotoxin (TTX) for 4--10 days, by implanting a glass capillary tube filled with TTX into the nerve through the epineurium. Following this treatment the somatotopic organization of receptive fields in the L4 dorsal horn, an area of cord normally responding only to foot stimulation. The map was normal in animals treated with TTX. Dorsal horn levels of fluoride-resistant acid phosphatase, substance P, somatostatin, cholecystokinin-like peptide, neurotensin and neurophysin were also normal as assessed from density of staining. These results are discussed in the light of the positive changes that are seen following chronic sciatic nerve section. PMID- 6287372 TI - Virilizing tumor of adrenal cortex in an infant. PMID- 6287373 TI - Amiodarone: its side effects, adverse reactions and dosage schedules. PMID- 6287374 TI - Resuscitation of the newborn. PMID- 6287375 TI - Cancer of the vulva. PMID- 6287376 TI - Reproductive wastage. PMID- 6287377 TI - [Cultivation of hepatitis A virus in tissue culture: the possibility of virus production for vaccines and testing, for the diagnosis of infection in patients and for the testing of disinfectants]. PMID- 6287378 TI - [Pathogenic microorganisms in the effluent from water purification plants and in the recipient basin]. PMID- 6287379 TI - Metastatic small cell carcinoma masquerading as orbital myositis. AB - A case of discrete metastases to an extraocular muscle from a silent small cell carcinoma is described. Clinically, diagnostically, and pathologically this masqueraded as orbital myositis. It was not until a full thickness muscle biopsy was obtained that the diagnosis was realized. The need for deep biopsy is stressed. Discrete metastases to an extraocular muscle without other orbital soft tissue involvement is rarely reported. This is the first reported case from a small cell primary tumor. Metastatic orbital disease is discussed. PMID- 6287380 TI - [Leukocyte migration inhibition elicited by Epstein-Barr virus antigen. Reaction potential in infectious mononucleosis as well as in states of immune deficiency and immunosuppression]. PMID- 6287381 TI - [Experience with varicella zoster immunoglobulin in Hungary]. PMID- 6287382 TI - [Adrenal cortex adenoma causing Cushing syndrome and virilization]. PMID- 6287383 TI - [Post-endodontic care]. PMID- 6287384 TI - Peripheral and central substrates involved in the rostrad transmission of nociceptive information. PMID- 6287385 TI - [Comparative study of ceftriaxone, cefotaxime and moxalactam against 150 Gram negative strains (author's transl)]. AB - The in vitro activity of ceftriaxone, a new cephalosporine derivative was compared with two other "third generation" cephalosporins, cefotaxime and moxalactam. 150 strains of Gram negative bacilli were used in this study. Ceftriaxone as cefotaxime and moxalactam were very active against most of the 104 multiresistant Enterobacteriaceae. Ceftriaxone was the most active against Proteus providencia with a mean MIC of 0.007 microgram/ml. Cefotaxime and ceftriaxone were less active than moxalactam against Enterobacter with MIC higher or equal to 32 micrograms/ml. The three cephalosporins had an almost identical activity against carbenicillin resistant Pseudomonas with an MIC higher than 4 micrograms/ml. MIC against Acinetobacter were higher or equal to 16 micrograms/ml. Moxalactam was the most active against Bacteroides fragilis with a mean MIC of 0.5 micrograms/ml; ceftriaxone and cefotaxime had a mean MIC of 2 to 4 micrograms/ml. PMID- 6287386 TI - [Human pharmacology of ceftriaxone (author's transl)]. AB - The human pharmacology of ceftriaxone was studied in five healthy adult subjects receiving a 500 mg dose intravenously. Evolution of serum levels represents a two compartment open model; the distribution volumes, central and peripheric, were low (about 3 L); the elimination half-life was particularly long : 7.87 hours; the renal and serum clearances and low. These data are probably related to the high affinity of the drug for serum proteins (95 p. cent bound); they suggest once daily parenteral dosing. PMID- 6287387 TI - [Clinical study of ceftriaxone (author's transl)]. PMID- 6287388 TI - [Clinical evaluation of cefotiam in adults urinary tract infections (author's transl)]. AB - Cefotiam, a new broad-spectrum cephalosporin was used for the treatment of 30 cases of urinary-tract infections (UTI). Patients, 20 females, 10 males were between 23 and 76 years old. UTI were 17 cystitis, 9 pyelonephritis and 4 prostatitis. Bacteria isolated from urine were : 24 E. Coli, 3 Proteus mirabilis, 1 Providencia, 3 Klebsiella, 3 Enterobacter, 2 Serratia, 1 Staphylococcus coagulase--, DNAse--. MIC's of cefotiam ranged from 0.003 to 32 micrograms/ml (median MIC : 0.06 microgram/ml). Cefotiam was given intramuscularly in monotherapy, at the daily dosage of 1 and 2 g in two injections during 7 to 28 days. Followup of patients were at least 4 weeks after the end of treatment. Therapeutic results were : 15 cures and 2 failures by relapse in 17 cystitis, 6 cures and 3 failures by relapse in 9 pyelonephritis, 4 cures in 4 prostatitis. General tolerance was excellent in all cases. Intramusculary injections were well tolerated with a 2 p. cent lidocain solution. Biological tolerance and particularly renal tolerance was good in all patients. PMID- 6287390 TI - Childhood hypertension due to adrenocortical disorders. PMID- 6287391 TI - Diagnosis and treatment of renal tubular disorders in children. PMID- 6287389 TI - [Statistical regression analysis for four new cephalosporins (author's transl)]. AB - The MIC (agar dilution method) and the zone diameter (disk diffusion) were determined for two hundred and sixteen of the most frequently bacterial strains found in hospitals in order to establish the regression curves for four new cephalosporins : cefotaxime, cefoperazone, cefotiam, moxalactam. The dta (MIC, zone size) were represented by a series of points (Mi (Xi Yi) in semilogarithmic rectangular co-ordinates expressed as : Xi = log2 MIC + 7 ; Yi = zone diameter. Firstly, we tried to fit a regression polynomial to this series of points progressing from degree 1, simple equation (standard regression line) 2, 3, etc. in order to reduce if need be, the deviation between the model (polynomial) and the empirical data. For these four cephalosporins a regression polynomial was obtained. Secondly, the global series was divided into three series of points Mi corresponding to three ranges of MIC : MIC less than or equal to 4 microgram/ml ; 4 less than or equal to 16 micrograms/ml; MIC greater than or equal to 16 microgram/ml. A regression line was obtained for each series and compared with each other and each one with the global regression line (they do not coincide statistically). Concerning these antibiotics, it is better to use a regression polynomial rather than apply a standard regression line. PMID- 6287392 TI - Vitamin D in bone disease. PMID- 6287394 TI - The effect of benzo(a)pyrene on fertility, primordial oocyte number, and ovarian response to pregnant mare's serum gonadotropin. AB - The polycyclic aromatic hydrocarbon (PAH) benzo(a)pyrene (BP) reduced the fertility of DBA/2N mice in a dose-dependent fashion. Control mice produced offspring at a rate of 0.91 pups/mouse per week. Treatment with BP at doses of 10, 100, 200, and 500 mg/kg decreased offspring production rates to 0.61, 0.20, zero and zero pups/mouse per week, respectively. BP also destroyed primordial oocytes in similarly treated mice. Treatment with BP at doses of 10, 50, 100, and 500 mg/kg destroyed 20%, 58%, 88%, and 100%, respectively, of the primordial oocytes in DBA/2N mouse ovaries. Dose-response curves for fertility reduction and primordial oocyte destruction were identical. The threshold for fertility reduction was 3.4 mg/kg and for primordial oocyte destruction it was 2.7 mg/kg. The 50% effect doses for fertility reduction and primordial oocyte destruction were 25.5 mg/kg and 24.5 mg/kg, respectively. Although BP reduced fertility and destroyed primordial oocytes, it had no effect on the ovarian weight response to pregnant mare's serum gonadotropin (PMSG), consistent with the observations that BP at these doses does not destroy growing or preovulatory oocytes or follicles. The similarity of the dose-response curves for fertility reduction and primordial oocyte destruction suggested that their mechanism of action is similar and resides within the ovary. As BP had no effect on ovarian response to PMSG, the effect was likely to reside outside the regulatory mechanisms controlling ovulation. These data suggest that the site of fertility reduction by BP may reside in the mechanisms of fertilization, implantation, or early conceptus development. PMID- 6287393 TI - Hepatic drug metabolizing enzyme activity and tumorigenesis in mice following perinatal exposure to benzo(a)pyrene. AB - It is known that in utero exposure to polycyclic aromatic hydrocarbon carcinogens (PAH) results in a high incidence of pulmonary and other tumors in the offspring, usually after a long latency period. The present study tested the hypothesis that perinatal exposure to benzo(a)pyrene (BP) produces alterations in microsomal mixed-function oxidase (MFO) activity that modify the response to postnatal challenge with a PAH in such a way as to render the offspring to be more susceptible to tumorigenesis, and to determine whether the gestational age at which BP exposure occurred was a determinant of the proposed alterations. Swiss mice were treated with BP on day 3, 8, 10, 12, 14, or 18 of gestation. Subgroups from each gestational-age treatment group and appropriate controls were studied for enzyme activity or were challenged with 3-methylcholanthrene (3-MC) when three months old. Female offspring of BP-treated mice had significantly elevated hepatic microsomal aminopyrine demethylase activity; cytochrome P-450 concentrations were significantly lower in male offspring. The most striking finding was the increased susceptibility to tumorigenesis of female offspring. Latency for local tumors was decreased by two weeks; tumors were more frequent and grew more rapidly. Significantly more female offspring had both local and pulmonary lesions, and the total number of pulmonary adenomas was also significantly higher. Further studies are required to determine if the developmental alterations in hepatic (MFO) activity resulting from perinatal exposure to PAH relate in a causative way to the enhanced susceptibility of female offspring to tumorigenesis. PMID- 6287395 TI - Study of the sensitivity of neonates to digoxin: contribution of erythrocyte 86rubidium uptake test. AB - In general, there is little agreement how digoxin should be used in newborn, and the results of studies in this field seem contradictory. This study attempts a quantitative assessment of the number and the sensitivity of cellular receptors for digoxin in the organism, by the in vitro measurement of erythrocyte 86Rubidium uptake in neonates compared with adults and old people. Red blood cells are first incubated with differing concentrations of digoxin, and then incubated with 86Rb. The initial level of 86Rb uptake (Rbi) is that observed in the absence of digoxin. The 50% index of captation (IC50) is the digoxin concentration in nanograms per ml at which 86Rb uptake is half Rbi. Three groups of patients were studied: Group I: 12 neonates, less than 5 days old; Group II: 11 adults (26 to 57 years old); Group III: 9 elderly people (71 to 82 years old). Rbi was significantly lower in neonates (Mean +/- SD: 25.8% +/- 3.5, P less than 0.001) and in the elderly (29.9% +/- 3.1) than in adults (36.8% +/- 4.6). IC50 was significantly lower in the elderly (12.1 ng/ml +/- 2.4) than in the adult patients (20.5 ng/ml +/- 5.5, P less than 0.001). In the newborns, values of IC50 were widely scattered (16.2 ng/ml +/- 7.2). The authors suggest that since Rbi reflects Na+, K+-ATPase activity, this activity is diminished in newborn and old people, and indicates that they have fewer cellular receptors for digoxin than adults. In the elderly, the low IC50 would imply increased sensitivity to digoxin. In neonates, the wide range of values for IC50 suggests considerable individual variation in sensitivity to digoxin. The results are consistent with the recently recommended lower dosages of digoxin in neonates. PMID- 6287396 TI - CT in the staging of children with malignant tumours. AB - The application of CT whole-body scanning as an adjunct in the staging investigations was evaluated in 56 children with various types of extra-cranial solid malignant tumours. CT proved to be superior to conventional films in the detection of pulmonary or pleural dissemination. The abdominal lymph nodes were difficult to evaluate by CT, and abnormal nodes could only be diagnosed when they were unequivocally enlarged. The role of CT in the detection of liver metastases could not be established in this study due to the rare occurrence of liver involvement. CT seemed to be a valuable method in the prediction of the operability of the primary tumour, but seemed to have an inability to show local invasiveness of the tumour. CT is an important supplement to conventional, noninvasive methods in the staging of these patients because CT is able to detect otherwise undetectable dissemination and this results in an improved therapeutic approach in individual cases. PMID- 6287397 TI - Pulmonary blastoma. PMID- 6287398 TI - The effect of route of delivery on neonatal natural killer cytotoxicity and antibody-dependent cellular cytotoxicity to herpes simplex virus-infected cells. AB - The ability of human neonatal and adult Ficoll-hypaque purified mononuclear cells to mediate natural killer cytotoxicity (NKC) and antibody-dependent cellular cytotoxicity (ADCC) against 51Cr labeled herpes simplex virus-infected (HSV infected) and uninfected cells was evaluated in healthy term infants delivered vaginally or by Cesarean (C)-section without labor, and in healthy adult controls. Cord blood NKC to HSV-infected cells (12.5 +/- 7.0) was lower (P less than 0.01) than adult controls NKC (29.5 +/- 7.0). NKC to HSV-infected cells of babies delivered vaginally (16.6 +/- 3.4) was lower (P less than 0.05) than adult controls (28.4 +/- 4.2). NKC to HSV-infected cells of neonates delivered by C section without labor (7.6 +/- 2.8) was also lower (P less than 0.001) than adult controls (30.7 +/- 4.0) and lower (P less than 0.05) than that of neonates delivered vaginally. Cord blood ADCC (43.1 +/- 9.0) was lower (P less than 0.05) than ADCC of adult controls (58 +/- 10). ADCC of neonates delivered vaginally (50 +/- 5.9) was similar to ADCC of adult controls (57.4 +/- 6.9). ADCC of neonates delivered by C-section without labor (30.4 +/- 7.2) was lower than ADCC of adult (58.5 +/- 7.4) and was lower (P less than 0.05) than ADCC of neonates delivered vaginally. These findings demonstrate that the method of delivery influences subsequent neonatal leukocyte NKC and ADCC. Further experiments will delineate the cause of these variations, which probably include labor or stress related hormonal changes in the mother or neonate. PMID- 6287399 TI - Lung development in the chick embryo. II. Choline and ethanolamine kinase activity in the developing lungs of normal and hypophysectomized chick embryos. PMID- 6287400 TI - Abnormal growth kinetics and 5'-nucleotidase activities in cultured skin fibroblasts from patients with Duchenne muscular dystrophy. PMID- 6287401 TI - Decreased neutrophil beta adrenergic receptors in the neonate. AB - Using cord blood neutrophils (PMN) the maturity of the beta adrenergic receptor has been assessed. [3H]-Dihydroalprenolol isoproterenol-induced cyclic AMP generation to characterize receptor function. Data from six neonates and six adult controls revealed that neonatal PMN had significantly (P less than 0.001) fewer receptor sites (728 +/- 43 receptors/cell) compared to the adult (1302 +/- 68 receptors/cel); however, the affinity constants (Kd) were very similar (0.78 +/- 0.14 nM compared to 0.61 +!- 0.05 nM). Neonatal PMN also generated less isoproterenol-induced cyclic AMP than adult cells at all concentrations, reaching significance (P less than 0.005) at 10(-4) - 10(-6) M isoproterenol concentrations. No difference in the isoproterenol concentrations stimulating 50% of the maximal cyclic AMP generation (EC50) was found between the two populations. These data indicate that the neonatal PMN has decreased beta adrenergic receptor sites. No evidence of a receptor--adenylate cyclase coupling defect was detected. These findings could reflect ontogenic differences between the neonate and adult PMN. The effect of increased catecholamine secretion during the stress of delivery resulting in 'down regulation' of the beta adrenergic receptor cannot presently be ruled out. PMID- 6287402 TI - The newborn of diabetic rat. I. Hormonal and metabolic changes in the postnatal period. PMID- 6287403 TI - [Prenatal infection with herpes virus]. PMID- 6287404 TI - [Diagnostic value of computed tomography in hepatocellular carcinomas (author's transl)]. PMID- 6287405 TI - [Utility of dynamic computed tomography in liver tumors (author's transl)]. PMID- 6287406 TI - [Effects of 70 Me V proton beam on a murine tumor and skin (author's transl)]. PMID- 6287407 TI - [Extention of the concept of antireceptor autoantibody disease]. PMID- 6287408 TI - [Is there a relationship between Epstein-Barr virus and rheumatoid arthritis (author's transl)]. AB - Antibodies directed against a nuclear antigen of lymphocytes infected by Epstein Barr virus (RANA antigen) have been detected in the sera of patients with rheumatoid arthritis, where they were present at higher levels than in the sera of controls. In vitro, the T lymphocytes of patients with rheumatoid arthritis seem to be incapable of controlling the multiplication of B lymphocytes induced by Epstein-Barr virus. The stimulated T lymphocytes produce more rheumatoid factors than normal lymphocytes. All this suggests that the Epstein-Barr virus might play a part in the pathogenesis of rheumatoid arthritis--a multifactorial disease which also involves a genetic factor, as shown by the increased prevalence of HLA DR4 antigen. PMID- 6287409 TI - [Pseudomembranous rectocolitis after treatment by cefotaxime]. PMID- 6287411 TI - [Male breast tumours. Anatomico-clinical study based on 73 cases (author's transl)]. AB - Breast cancer is 100 times less frequent in males than in femalees. In this series the predominant histological form clearly was adenocarcinoma, present in 69 out of 73 patients. The patients' age varied from 7 to 82 years. In 91% of the cases, the malignancy presented as a painless, isolated tumour, often associated with retraction of the nipple. After 5 years 68% of patients operated upon immediately after diagnosis and 35% of those initially treated by radiations were still alive, as were 72% of patients with less than 7 cm tumours and 44.8% of those with larger tumours. It appears that breast cancer in males is less aggressive than is usually believed. Its histological features and its treatment are comparable to those of femal breast cancer. PMID- 6287410 TI - [Neonatal pneumopathy due to para-influenza 1 virus. Identification by bronchoalveolar lavage]. PMID- 6287412 TI - [A rare cause of hypercalcaemia: familial hypocalciuric hypercalcaemia (author's transl)]. AB - Two cases of familial hypocalciuric hypercalcaemia (FHH) in the same family are presented. This congenital disease is characterized by biochemical evidence of parathyroid toxicity but differs from primary hyperparathyroidism by the presence of normal parathyroid hormone levels. However, histomorphometric studies of transiliac bone biopsy specimens after double-labeling with tetracycline shows intensive remodelling of bone tissue with increase in osteoclastic resorption and osteoblastic apposition areas, all features identical with those observed primary hyperparathyroidism. The authors suggest hypersensitivity of target-organs (bones and kidneys) to parathyroid hormone as a probable physiopathological mechanism for FHH. PMID- 6287413 TI - [Hepatocellular carcinoma in an African after 10 years of treatment of Waldenstrom's disease with chlorambucil]. PMID- 6287415 TI - Initiation of transcription of genes for mitochondrial ribosomal RNA in yeast: comparison of the nucleotide sequence around the 5'-ends of both genes reveals a homologous stretch of 17 nucleotides. AB - The DNA sequence around the beginning of the genes coding for the large and small ribosomal RNAs in yeast mitochondria has been established. In order to determine the 5'-end points of the ribosomal RNAs, DNA fragments were labelled in vitro at a restriction site within each gene and hybridized with ribosomal RNA. The hybrids were then treated with S1 nuclease and the products analysed for size by gel electrophoresis. This enabled us to identify where in the determined DNA sequence the 21S ribosomal RNA and the precursor for 15S ribosomal RNA (15.5S rRNA) start, since both transcripts are initiated de novo (Levens et al. (1981) J.Biol.Chem., 256, 5226-5232). Comparison of the DNA sequences around the start points of transcription reveals the existence of a homologous stretch of 17 nucleotides. This conserved sequence may be an essential element of a promoter in mtDNA. PMID- 6287414 TI - RNA splicing is required to make the messenger RNA for a variant surface antigen in trypanosomes. AB - The expression of the gene for variant surface glycoprotein (VSG) 118 in Trypanosoma brucei is activated by transposing a DNA segment containing the gene and 1-2 kb in front of it to an expression site elsewhere in the genome. By S1 nuclease protection and RNA blotting experiments we show here the presence of several minor transcripts in trypanosomes synthesizing VSG 118, one of which covers the entire transposed segment. Comparison of the sequence of the 5' terminal segment of VSG 118 messenger RNA (mRNA), determined by primed reverse transcription, and the corresponding region of the 118 VSG gene, shows that the 5' terminal 34 nucleotides of the mRNA are not encoded in the 118 VSG gene contiguous with the remainder of the mRNA. We conclude that synthesis of a VSG mRNA involves splicing of a much longer primary transcript, which may start outside the transposed segment. PMID- 6287416 TI - Differences in the nuclease sensitivity between the two alleles of the immunoglobulin kappa light chain genes in mouse liver and myeloma nuclei. AB - In mouse myeloma T the productive kappa light chain gene differs from its aberrantly rearranged allele in the patterns of DNAase I hypersensitive sites. In the region of the alleles where they are identical in sequence they have one site in common which lies 0.8 kb downstream of the coding region; but two sites upstream of and within the C gene segment (2) are found only on the non productive allele. Within the region of different sequences both alleles have analogously located DNAase I hypersensitive sites; they lie 0.15 kb upstream of the respective leader segments and cover putative promoter sequences. Only one of the six DNAase I hypersensitive sites is also very sensitive towards micrococcal nuclease due to its particular DNA sequence. The non-rearranged gene studied in liver nuclei has no DNAase I hypersensitive sites but is preferentially cleaved in A/T rich regions. PMID- 6287417 TI - Site directed in-vitro assembly of nucleosomes. AB - An in-vitro system is described allowing for the assembly of nucleosomes on preselected sites of a cloned tRNA gene. The system consists of a soluble nucleoprotein fraction, a histone source, and circular DNA containing a single stranded stretch (sssDNA). Nucleosomes assemble on the sssDNA, if the three components are incubated as a highly concentrated solution in the presence of the four deoxyribonucleotidetriphosphates and of ATP. The single stranded stretch is rendered double stranded during incubation. The middle axis of one assembled nucleosome always coincides roughly with the midpoint of the original single stranded DNA stretch. PMID- 6287418 TI - Nucleotide sequence of the region between the 18S rRNA sequence and the 28S rRNA sequence of rat ribosomal DNA. AB - The DNA sequence of the intragenic region of the rat 45S ribosomal RNA precursor was determined. This sequence contains 2282 nucleotides and extends from the conserved EcoR I site near the 3' terminus of 18S rRNA to 69 nucleotides downstream of the 5' terminus of 28S rRNA. The sequences corresponding to 18S and 5.8S rRNA were identified by comparison with previously published data. The 5' terminus of rat 28S rRNA was identified by S1 nuclease protection and reverse transcriptase elongation assays. The internal transcribed spacers were found to be 1066 and 765 nucleotides long and had little homology with those of Xenopus and yeast. Regions of sequence homology between rat and Xenopus were found at the junctions of the internal transcribed spacers with 18S, 5.8S and 28S rRNA. These homologies suggest that these sequences may function as recognition sites for the processing of the ribosomal precursor RNA. PMID- 6287419 TI - Enzymatic replacement in vitro of the first anticodon base of yeast tRNAAsp: application to the study of tRNA maturation in vivo, after microinjection into frog oocytes. AB - A combination of several enzymes, RNase-T1, nuclease S1, T4-polynucleotide kinase and T4-RNA ligase were used to prepare and modify different fragments of yeast tRNAAsp (normal anticodon G U C). This allowed us to reconstitute, in vitro, a chimeric tRNA that has any of the four bases G, A, U or C, as the first anticodon nucleotide, labelled with (32p) in its 3' position. Such reconstituted (32p) labelled yeast tRNAAsp were microinjected into the cytoplasm or the nucleus of the frog oocyte and checked for their stability as well as for their potential to work as a substrate for the maturation (modifying) enzymes under in vivo conditions. Our results indicate that the chimeric yeast tRNAsAsp were quite stable inside the frog oocyte. Also, the G34 was effectively transformed inside the cytoplasm of frog oocyte into Q34 and mannosyl-Q34; U34 into mcm5s2U and mcm5U. In contrast, C34 and A34 were not transformed at all neither in the cytoplasm nor in the nucleus of the frog oocyte. The above procedure constitutes a new approach in order to detect the presence of a given modifying enzyme inside the frog oocyte; also it provides informations about its cellular location and possibility about its specificity of interaction with foreign tRNA. PMID- 6287420 TI - Structure of replication origin of the Escherichia coli K-12 chromosome: the presence of spacer sequences in the ori region carrying information for autonomous replication. AB - The replication origin region of the Escherichia coli K-12 chromosome has been cloned, and a region of 245 base-pairs has been shown to contain all the information for autonomous replication (defined ori). In order to obtain further information on the sequence organization in the defined ori stretch, various types of mutation were introduced by in vitro techniques at a restriction site (AvaII site) which locates near the middle of ori. When the correlation between these mutations and replicating function was examined, different effects were obtained with the types of mutation: the replicating function was completely destroyed by either insertion or deletion of short sequences, but not by base substitutions. Based on these observations and on the fact that multi-gene products are involved in the initiation of replication, we assume that two categories of sequences are present within the ori stretch, one specifying interaction with initiation factors (recognition sequences) and the other spacing the recognition sequences in appropriate distances (spacer sequences), and that the AvaII site is located in the spacer region. PMID- 6287421 TI - Expression of Drosophila heat shock genes is regulated in Rat-cells. AB - We have introduced a Drosophila 87A heat shock locus into the genome of Rat-1 cells. A brief heat shock of a resultant transformed line, Rhs11, induces the transcription of the Drosophila heat shock genes from the Drosophila heat shock promoter. This suggests that the induction mechanism and regulatory sequences for the heat shock response have been conserved between Drosophila and mammals. PMID- 6287422 TI - Nucleotide sequence of a chromosomal rearranged lambda 2 immunoglobulin gene of mouse. AB - The rearranged lambda 2 gene of the mouse plasmacytoma cell line MOPC315 has been cloned and sequenced. A comparison of its sequence with the sequence of the unrearranged (germ-line) V, J and C gene segments shows that the sequences of the V gene segments differ at six positions. The sequence of the J and C gene segments remained unchanged. These results add support to the hypothesis that somatic mutations occur in immunoglobulin in genes and that these mutations do not involve the C gene segment. The degree of homology of the elements of the lambda 2 gene with those of the lambda 1 gene and C lambda 3 and C lambda 4 gene fragments suggest a pathway of evolution by gene duplication of the immunoglobulin lambda light chain locus. According to this scheme the original structure V0-J0C0 gave rise to a structure V0-J1C1-J11C11 by duplication of the J0C0 region. A second duplication encompassing the whole region resulted in the present structure: V1-J3C3-J1C1/V2-J2C2-J4C4. PMID- 6287423 TI - Isolation and characterization of maize genes coding for zein proteins of the 21000 dalton size class. AB - Cloning in lambda gt WES phage of EcoRI fragments from maize seedlings DNA led to the isolation of four fragments containing genes coding for 21000 dalton zein proteins. The zein genes, identified by electron microscopic analysis, do not contain intervening sequences detectable by this method. The flanking sequences were analyzed by restriction sites mapping and hybridization and showed areas of homology between each other and with sequences surrounding a previously isolated gene of the 19000 dalton zein class. PMID- 6287424 TI - The complete nucleotide sequence of the chick a-actin gene and its evolutionary relationship to the actin gene family. AB - The nucleotide sequence of the chick a-actin gene reveals that the gene is comprised of 7 exons separated by six very short intervening sequences (IVS). The first IVS interrupts the 73 nucleotide 5' untranslated segment between nucleotides 61 and 62. The remaining IVS interrupt the translated region at codons 41/42, 150, 204, 267, and 327/328. The 272 nucleotide 3' untranslated segment is not interrupted by IVS. The amino acid sequence derived from the nucleotide sequence is identical to the published sequence for chick a-actin except for the presence of a met-cys dipeptide at the amino-terminus. The IVS positions in the chick a-actin gene are identical to those of the rat a-actin gene. While there is partial coincidence of the IVS in the a-actin genes with the vertebrate b-actin genes and 2 sea urchin actin genes, there is no coincidence with actin genes from any other source except soybean where one IVS position is shared. This discordance in IVS positions makes the actin gene family unique among the eucaryotic genes analyzed to date. PMID- 6287425 TI - Transcriptional analyses of the Bacillus licheniformis penP gene. AB - We report the promoter structure of the Bacillus licheniformis 749/C penicillinase (penP) gene. The transcript encoding the penicillinase gene was identified by in vitro run-off transcription using both E. coli RNA polymerase and B. subtilis RNA polymerase. Utilization of this promoter in linearized DNA by the B. subtilis RNA polymerase showed extreme sensitivity to ionic strength. The 5' sequence of the penP mRNA was determined using enzymatic sequencing (1). Holoenzymes from E. coli and B. subtilis (E sigma 55) initiate penP RNA synthesis at the same site. Alignment of this RNA sequence with the reported DNA sequence of ther penP gene (2,3) revealed the "-35 region" and "Pribnow box" sequences that are recognized as 5'TTGCAT and 5'AATACT, respectively. Potential secondary structure within the promoter exists which may play a role in the expression of the penicillinase gene. The location of the penP promoter was further confirmed by molecular cloning of a DNA fragment containing the expected promoter sequence into a promoter-fishing vector useful for monitoring promoter activities in both E. coli and B. subtilis. PMID- 6287426 TI - Identification of the in vivo and in vitro origin of transcription in human rDNA. AB - A Hela cell /-100 extract primed with a purified human rDNA containing clone, has been shown to be capable of initiating specific alpha-amanitin-resistant RNA transcripts. By using a number of truncated templates, the site of RNA polymerase I initiation in vitro has been identified. The origin of transcription in vitro and in vivo was further defined by S1-mapping studies with total Hela cell RNA or RNA isolated from the in vitro transcription reaction. The initiation site was found to be the same. The nucleotide sequence of an 848 bp region around the initiation site, has also been determined. A perfect 15 bp homology has been found to exist between human and mouse rDNA very close to the origin of transcription, although little homology exists elsewhere. Sequences homologous to the origin of transcription region were not found repeated within a 12 kb non transcribed spacer segment upstream from it. PMID- 6287428 TI - The changes in proviral chromatin that accompany morphological variation in avian sarcoma virus-infected rat cells. AB - The clone All of avian sarcoma virus B77-infected Rat-1 cells comprises both morphologically normal and morphologically transformed derivatives. Transformed subclones, in which virus-specific RNA is readily detectable, contain a provirus that is very sensitive to DNase 1 digestion of chromatin, and show DNase 1 hypersensitive sites at the 5' end of the provirus and in 5' flanking cell DNA. Normal subclones with no detectable virus-specific RNA, whether infected cells that have never been transformed or revertants derived from transformed cells, contain a provirus that is far more resistant to DNase 1 digestion. Moreover this provirus lacks hypersensitive sites at its 5' end, although DNase 1 hypersensitive sites were detected at the 3' end of the provirus in either normal or transformed clones. The pattern of cytosine methylation in the proviral restriction sites of the isoschizomers Msp I and Hpa II differed between transformed and revertant clones; the revertants show additional methylation at some CpG doublets. PMID- 6287427 TI - Deoxyribonuclease II as a probe to sequence-specific chromatin organization: preferential cleavage in the 72 bp modulator sequence of SV40 minichromosome. AB - T prove sequence-specific chromatin structure of SV40 minichromosome, we further modified previously described hybridization mapping. Actually (i) the digestion patterns by two nucleases (micrococcal and DNAase II) were compared and (ii) the kinetics of nuclease digestion was analyzed from early time points when only a fraction of minichromosomes was cleaved once to longer digestions when oligo- and mononucleosomal bands appeared. DNAase II is shown to possess certain sequence specificity different from that of micrococcal nuclease. The major finding is that DNAase II preferentially cleaves the SV40 minichromosome at a distinct region of the genome known as 72 bp modulator element. Other hypersensitive sites are located near the replication origin and T-ag binding site II and also near BamHI site where termination of replication and "late" transcription occurs. Micrococcal nuclease splits the BglI-Hpaii region in a different manner. PMID- 6287429 TI - Insertion sites and the terminal nucleotide sequences of the Tn4 transposon. AB - The nucleotide sequences at the ends of the Tn4 transposon (mercury spectinomycin and sulfonamide resistance) have been determined. They are inverted repeated sequences of 38 nucleotides with three mismatched base pairs. These sequences are strongly homologous with the terminal sequences of Tn501 (mercury resistance) but less so with those of Tn3 (ampicillin resistance). The Tn4 transposon generates pentanucleotide members (Tn3, Tn1000, Tn501, Tn551, IS2) with the exception of Tn1721 and bacteriophage Mu. Among the three Tn4 insertion sites examined here, two of them occurred near a nonanucleotide sequence in perfect homology with part of the terminal inverted-repeat sequence of Tn4 and the third insertion occurred near a sequence of partial homology to one end of Tn4. All three insertions were in the same orientation such that IRb is proximal to its homologous sequence on the recipient DNA. PMID- 6287430 TI - The primary structure of E. coli RNA polymerase, Nucleotide sequence of the rpoC gene and amino acid sequence of the beta'-subunit. AB - The primary structure of the E. coli rpoC gene (5321 base pairs) coding the beta' subunit of RNA polymerase as well as its adjacent segment have been determined. The structure analysis of the peptides obtained by cleavage of the protein with cyanogen bromide and trypsin has confirmed the amino acid sequence of the beta' subunit deduced from the nucleotide sequence analysis. The beta'-subunit of E. coli RNA polymerase contains 1407 amino acid residues. Its translation is initiated by codon GUG and terminated by codon TAA. It has been detected that the sequence following the terminating codon is strikingly homologous to known sequences of rho-independent terminators. PMID- 6287431 TI - Synthesis and physicochemical properties of two analogs of poly(dA): poly(2 aminopurine-9-beta-D-deoxyribonucleotide) and poly 2-amino-deoxyadenylic acid. AB - Polymerization of chemically synthesized dn2h6ATP and dn2ATP by deoxynucleotidyl transferase from calf thymus furnished poly(dn2h6A) and poly(dn2A) respectively. The synthetic polynucleotides were characterized by spectroscopic, ultracentrifugation and enzymatic methods. In polynucleotide-polynucleotide interaction, poly(dn2h6A) and poly(dn2A) behaved like analogs of poly(dnA). PMID- 6287432 TI - The nucleotide sequence of a human immunoglobulin C gamma1 gene. AB - We report the nucleotide sequence of a gene encoding the constant region of a human immunoglobulin gamma 1 heavy chain (C gamma 1). A comparison of this sequence with those of the C gamma 2 and C gamma 4 genes reveals that these three human C gamma genes share considerable homology in both coding and noncoding regions. The nucleotide sequence differences indicate that these genes diverged from one another approximately 608 million years ago. An examination of hinge exons shows that these coding regions have evolved more rapidly than any other areas of the C gamma genes in terms of both base substitution and deletion/insertion events. Coding sequence diversity also is observed in areas of CH domains which border the hinge. PMID- 6287434 TI - [Anti-HA antibodies in a sample of an apparently healthy population of the province of Sassari]. PMID- 6287433 TI - Chromosome-bound mitotic factors: release by endonucleases. AB - Additional evidence is presented to support our recently reported conclusion that the mitotic factors of mammalian cells, which induce germinal vesicle breakdown and chromosome condensation when injected into fully grown Xenopus laevis oocytes, are localized on metaphase chromosomes. Chromosomes isolated from mitotic HeLa cells were further purified on sucrose gradients and digested for varying periods with either the micrococcal nuclease or DNase II. At each time point of digestion the amount of mitotic factors released was determined by injecting a supernatant of these fractions, obtained by high-speed centrifugation, into oocytes. The amount of DNA rendered acid soluble under the conditions of digestion used was 3% ot 5% of the total chromosomal DNA. The extent of release of mitotic factors with both nucleases was estimated to be about 30% to 40% as evidenced by the reextraction of the undigested chromosomal pellet with 0.2 M NaC1. Similar results were obtained when nuclei from G2 cells were digested under identical conditions. The release of these chromosome-bound mitotic factors by mild digestion with these nucleases though only partial, clearly demonstrates that a significant proportion of these factors are localized on metaphase chromosomes. PMID- 6287435 TI - A method of studying metabolic variation between individual tumors: a preliminary report. PMID- 6287436 TI - Carcinogens occurring naturally in food. AB - Carcinogens naturally occurring in food include metabolites of microorganisms which infect plants during harvesting and storage; products of the normal metabolism of plants and products of chemical changes occurring during processing, preservation, preparation or which are picked up from the environment, including contaminated water or soil. Some examples of these types of carcinogens are: fungal metabolites such as aflatoxins in peanuts stored under improper conditions; safrol from the oil of various plants; tannin in tea, grain and grapes; and polycyclic hydrocarbons including benzo (a) pyrene formed by smoking meat and fish. Epidemiological studies indicate that there is a high incidence of gastric cancer in the areas of the world where smoked fish are common in diets. Vitamin A may play a role in preventing the carcinogenic action of polycyclic hydrocarbons. PMID- 6287437 TI - Diet and cancer of the large bowel. AB - The incidence of large bowel cancer correlates internationally with the consumption of fat, protein or meat. Etiological hypotheses have been proposed, suggesting that a high-fat diet is related to an increased risk of large bowel cancer, while dietary fiber is claimed to have a modifying effect. However, while these hypotheses were derived from the analysis of the geographical distribution of colon cancer in humans, they are not unequivocally supported by the results of further epidemiological studies. This is particularly true for case-control studies which generally have not confirmed an association between diet and the risk of colorectal cancer at the individual level. Lack of uniformity among the findings of epidemiological studies concerned with the role of diet in the etiology of bowel cancer could be explained by 1. lack of sensitive and reproducible methods for assessment of diet in epidemiological studies, 2. lack of knowledge of the mode of action of dietary factors responsible either for the development of the tumor or for protection against it. Future epidemiological research on diet and large bowel cancer should deal with more precisely defined hypotheses in which the possible multifactorial nature of this tumor would be taken into account. PMID- 6287438 TI - Neural pathways. PMID- 6287439 TI - The phenomenon of cancer cachexia: a review. PMID- 6287440 TI - Extravasation management. PMID- 6287441 TI - [New case of extrarenal Wilms' tumor]. PMID- 6287442 TI - [Osteodystrophy in children with chronic renal insufficiency in dialysis therapy]. AB - One of the most important complication of patients with chronic renal failure is osteodystrophy. This causes skeletal deformities, growth failure, bone pain and decreased physical activity. Osteodystrophy is more frequent among children than uraemic adults. In fact, 50-80% of children with chronic renal failure may occur in metabolic bone disease and the incidence tends to be higher in those children who have been in uraemic state for a long time before starting chronic haemodialysis. Osteodystrophy is a result of: 1) lesions of rickets; 2) lesions of osteitis fibrosa: 3) osteosclerosis. In contrast to adult, metastatic calcifications are virtually never observed in uraemic children. Hyperphosphoraemia, that is secondary to the reduction of G.F.R., may be the principal responsible of hyperparathyroidism that is the main cause of osteodystrophy. Hyperparathyroidism is also maintained and increased by deficit of 1,25(OH)2D3 which is responsible for lesions of rickets. Haemodialysis may markedly improve osteitis fibrosa and it is efficacious in reversing the mineral defect. Dialysate calcium concentration should be maintained at approximately 3,5 mEq/l. In this case we can raise serum calcium. On the contrary dialysate has to be lacking in phosphorus to correct hyperphosphoraemia. It must be noted that we have to prepare a dialysate with deionized water lacking in aluminum to avoid encephalopathy compliance. PMID- 6287443 TI - [Serum 5' nucleotidase and alkaline phosphatase activity in acute viral hepatitis]. PMID- 6287444 TI - [Role of prostaglandins in thyroid function]. PMID- 6287445 TI - Neurological manifestations of paranasal disease: problems in prognosis. PMID- 6287446 TI - Studies of avian leukosis virus infection in chickens from a commercial breeder flock. AB - Progeny chicks from a commercial breeder flock with a high rate of avian leukosis virus (ALV) infection were classified as congenitally or contact infected with ALV on the basis of virus tests on their meconia at hatching and the presence or absence of antibodies at 20 weeks of age. The ALV infection profile, shedding, tumor mortality, and egg production in the congenitally-infected chickens were compared with those in the contact-infected hatchmates. Results show that viremic tolerant hens classed as congenitally infected consistently shed virus into albumen of their eggs and progeny and 18% died from tumors. In contrast, hens classified as contact infected had a transient viremia; most of them developed antibody to ALV, and only 10% shed virus into albumen and progeny. No mortality from tumors occurred in this group of chickens during the 48-week experimental period, and egg production was significantly higher than in the group classified as congenitally infected. In addition, progeny chickens hatched from the contact infected group maintained a low rate (2.9 to 3.8%) of ALV infection for the next two generations without further selection. These data further document principles and procedures for reduction of ALV infection in commercial flocks and the considerable economic benefits to be derived therefrom. PMID- 6287447 TI - Increased severity and new symptoms of infectious bursal disease during aflatoxicosis in broiler chickens. AB - A factorial design for the presence and absence of infection with a mild strain of infectious bursal disease (IBD) virus and graded levels of dietary aflatoxin (0, .625, 1.25, 2.5, 5.0, and 10.0 microgram/g of diet) in young broiler chickens revealed several interactions which greatly changed and character of the IBD infection. An infection that was essentially free of mortality became lethal to about half the birds. A coagulopathy as indicated by prolonged prothrombin times and by the occurrence of slight hemorrhages in skeletal muscles, a hypoproteinemia, a hypocalcemia, and a markedly enlarged kidney were observed in combined aflatoxicosis and IBD, but not in IBD alone. The size of the bursa of Fabricius following an initial hypertrophy was atrophied 4 days after inoculation and the spleen was enlarged. The effects of aflatoxin and IBD on these two glands were additive. Packed blood cell volume and hemoglobin values were decreased by aflatoxin only. Thus, aflatoxin had the effects of making IBD a much more severe disease and of changing the symptoms. PMID- 6287448 TI - [Cytostatic treatment of inoperable and after care of operable bronchial cancer]. PMID- 6287449 TI - [The problem of extracranial metastasis of malignant gliomas. Report of 2 cases and review of the literature]. PMID- 6287450 TI - [Hemostatic properties of thrombocytes and their cyclic 3,5-adenosine monophosphate and prostaglandin E2 content during the storage of preserved blood at 4 degrees C]. PMID- 6287451 TI - Activation of alloimmune memory cells by soluble factors: a model for elicitation of specific allograft responses by immunologically nonspecific signals. PMID- 6287452 TI - Hemodialysis with base free dialysate. PMID- 6287453 TI - Growth factor activation of an amiloride-sensitive Na+/H+ exchange system in quiescent fibroblasts: coupling to ribosomal protein S6 phosphorylation. AB - Chinese hamster lung fibroblast cells (CCl39) enter the G0/G1 nonproliferative state after serum deprivation. In this report, we show that reinitiation of DNA synthesis by serum or the combination of purified human thrombin and insulin (1 10 microgram/ml) is preceded by very early stimulation of ionic fluxes (Na+/Rb+) and protein phosphorylation (27,000 daltons, 62,000 daltons, and the ribosomal S6 proteins). The potentiating action of insulin on thrombin-stimulated DNA synthesis is also observed on thrombin-stimulated Na+ influx, Rb+ influx, and protein S6 phosphorylation. Moreover, we demonstrate that CCl39 cells possess a Na+/H+ exchange system sensitive to amiloride. Half-maximal inhibition of growth factor-activated Na+ influx and Na+-dependent H+ efflux is obtained with 3-10 microM amiloride. Two lines of evidence indicate that the extrusion of H+ via the activation of the Na+/H+ exchanger is coupled to protein S6 phosphorylation: serum-stimulated phosphorylation is blocked by (i) amiloride at a concentration that abolishes serum-stimulated Na+ influx and (ii) protonophores that acidify the cell interior. The present data support the idea that the regulation of intracellular pH is a key event in the mechanism of growth factor action. PMID- 6287454 TI - Marking the polarity of RNA molecules for electron microscopy by covalent attachment of psoralen-DNA restriction fragments. AB - We have developed a general procedure for covalently attaching complementary restriction fragments to RNA molecules and visualizing the fragment in the electron microscope after denaturation. This is accomplished by creating psoralen monoadducts on the restriction fragment by irradiation at 390 nm before hybridization to the RNA and subsequently converting the monoadducts to RNA . DNA crosslinks by irradiation at 360 nm. Using this procedure, we have determined the location of a crosslink placed in Escherichia coli 165 rRNA by a previous irradiation. A consistent assignment is obtained by marking the crosslinked RNA with restriction fragments complementary to the 5' end or the 3' end of the 165 rRNA molecule. PMID- 6287455 TI - Variability of DNA methylation patterns during serial passage of human diploid fibroblasts. AB - We have examined DNA methylation in diploid human fibroblasts, early and late in their replicative life-span. The extent of methylation of -C-C-G-G- was measured by comparison of fragment sizes after digestion with methylation-specific restriction enzyme Hpa II or Msp I, or both. Methylation of -C-C-G-G- sites in total DNA, occurring predominantly at internal (3') cytosines, increased from 59% to 64% of sites in one cell strain at late passage, remained constant in another, and decreased in four other strains (54% to 48%, 58.5% to 48%, 55% to 51.5%, and 52% to 44.5%). Base composition analysis confirmed a substantial loss of total DNA 5-methylcytosine (mC) in one strain. Seven clonal isolates, examined at middle to late passage, ranged from 33% to 51% methylation of 3' cytosines in -C C-G-G- sites. Three discrete classes of highly repetitive DNA were found which contained Msp I sites at intervals of 45, 110, and 175 base pairs. These repeat families consistently had 70-80% of sites methylated at 3' cytosines, in all clones and in all strains examined both at early and at late passage. Thus, altered methylation of repetitive sequences is unlikely to account for the variable -C-C-G-G- methylation observed in total DNA. When DNA from one fibroblast strain and from eight pure clones isolated from that parental culture was digested with Msp I or Hpa II followed by EcoRI and probed for gamma-globin gene sequences, considerable interclonal and intraclonal heterogeneity was observed for methylation at four -C-C-G-G- sites in the gamma-globin coding region of DNA. Therefore, the pattern of methylation in endogenous gene regions appears to undergo random drift during replication of diploid fibroblasts. PMID- 6287456 TI - Proteins specified by avian erythroblastosis virus: coding region localization and identification of a previously undetected erb-B polypeptide. AB - Avian erythroblastosis virus (AEV) induces erythroblastosis and sarcomas in chickens. Two domains within the viral genome, erb-A and erb-B, have been implicated in AEV-mediated oncogenesis. By use of hybridization-arrested translations and hybridization-selections of mRNA, we have mapped on the viral genome the polypeptides specified by the erb domains. The results of hybridization-arrest with DNA representing the spliced 5' leader region of the AEV mRNA suggested that the authentic product of the erb-B domain was a 61,000 molecular weight protein, not a 41,000 molecular weight polypeptide previously identified. PMID- 6287457 TI - Proteolytic processing of poliovirus polypeptides: antibodies to polypeptide P3 7c inhibit cleavage at glutamine-glycine pairs. AB - Proteolytic processing of poliovirus polypeptides was examined by the addition of antibodies directed against the viral proteins P3-7c and P2-X to a cell-free translation extract prepared from infected HeLa cells. Antisera to P3-7c specifically inhibited in vitro processing at Gln-Gly pairs. Partial amino acid sequence analysis revealed a second Tyr-Gly pair that is utilized in protein processing. Neither Tyr-Gly cleavage is affected by antibody to P3-7c. Anti-P3-7c antibodies react not only with P3-7c but also with P3-6a and P3-2, two viral polypeptides NH2-coterminal with P3-7c. Preimmune and anti-P2-X antibodies had no effect on the processing of poliovirus proteins in vitro. We conclude that the activity responsible for processing poliovirus polypeptides at Gln-Gly pairs resides in the primary structure of P3-7c (or P3-2 and P3-6a) and not in P2-X. PMID- 6287458 TI - Catabolism of low density lipoproteins by perfused rabbit livers: cholestyramine promotes receptor-dependent hepatic catabolism of low density lipoproteins. AB - Rabbits fed a wheat starch/casein diet develop a marked hypercholesterolemia accompanied by a decrease in the number of EDTA-sensitive binding sites on plasma membrane fractions of the liver for low density lipoproteins (LDL) and beta migrating very low density lipoproteins [Chao, Y.-S., Yamin, T.-T. & Alberts, A. W. (1982) J. Biol. Chem., in press]. Inclusion of 1% cholestyramine resin in this diet prevents the increase in plasma cholesterol, increases the removal of LDL from plasma, and increases the number of hepatic plasma membrane LDL-binding sites. To determine the functional role of hepatic LDL-binding sites in the catabolism of LDL, we studied the catabolism of (125)I-labeled LDL ((125)I-LDL) by in situ perfused rabbit livers in a recirculating system. The rate of catabolism was measured from the increment of nonprotein-bound radioiodine in the perfusate. The receptor-dependent catabolism of LDL by the liver was calculated from the difference of hepatic catabolism of (125)I-LDL and catabolism of (125)I labeled cyclohexanedione-modified LDL, which does not bind to LDL receptors. The data show that about 74% of LDL catabolized by perfused livers from chow-fed rabbits is through the receptor-dependent pathway and 26% is through the receptor independent pathway. In rabbits fed a cholesterol diet, the hepatic catabolism of (125)I-LDL is reduced, and the receptor-dependent catabolism of (125)I-LDL is abolished. In rabbits fed the wheat starch/casein diet, the receptor-dependent catabolism of (125)I-LDL is reduced by 40% when compared with hepatic catabolism in chow-fed rabbits. Perfused livers from rabbits fed the wheat starch/casein diet supplemented with 1% cholestyramine show a 5,4-fold increase of receptor dependent catabolism of (125)I-LDL when compared with that of livers from rabbits fed the wheat starch/casein diet alone. Thus, these studies demonstrate that the change in the number of rabbit hepatic membrane LDL receptors induced by dietary manipulation and drugs is correlated to the functional rate of removal of LDL by the liver. PMID- 6287459 TI - Asymmetric replication of duck hepatitis B virus DNA in liver cells: Free minus strand DNA. AB - In order to study the replication of the DNA genome of duck hepatitis B virus, an avian virus related to human hepatitis B virus, we have characterized viral DNAs present in the livers of viremic ducks by agarose gel electrophoresis and the Southern blot procedure. In addition to relaxed circular DNA similar to virion DNA, livers contained a heterogeneous population of rapidly migrating species. The conformation of the rapidly migrating species was markedly sensitive to salt, suggesting that these species were largely single stranded. The largest major rapidly migrating species was shown to have an electrophoretic mobility that was insensitive to preheating of the DNA to 100 degrees C and was similar to that of denatured virus DNA 3 kilobases long, suggesting that this DNA was a single stranded copy of the entire virus genome. Hybridization with strand-specific probes demonstrated that this 3-kilobase species, as well as more rapidly migrating DNAs, were predominantly minus strands. PMID- 6287460 TI - Involvement of outside DNA sequences in the major kinetic path by which EcoRI endonuclease locates and leaves its recognition sequence. AB - We have examined the kinetics of the interaction between endodeoxyribonuclease EcoRI (EC 3.1.23.13) and nine linear DNA fragments that range in size between 34 and 6,200 base pairs and contain the EcoRI site of plasmid pBR322 in a central location. The kinetic parameters governing both formation and decay of specific endonuclease . DNA complexes increase 8-fold with increasing chain length over this size range. In contrast, equilibrium competition experiments demonstrated that the intrinsic affinity of endonuclease for its recognition sequence is independent of DNA chain length over this range. Thus, DNA sequences outside the recognition site enhance the rate at which EcoRI endonuclease locates or leaves its recognition site without affecting the intrinsic thermodynamic parameters of site-specific interaction. These results are consistent with a facilitated diffusion mechanism for specific DNA site location by this enzyme. PMID- 6287461 TI - Purification of polyoma virus medium-size tumor antigen by immunoaffinity chromatography. AB - We have used antibodies against the synthetic peptide Lys-Arg-Ser-Arg-His-Phe, corresponding to the six COOH-terminal amino acids of the polyoma virus medium tumor (T) antigen, to purify the medium T antigen by affinity chromatography. Release of the medium T antigen from the anti-peptide antibody was achieved under mild conditions by using a large excess of the peptide in an isotonic buffer at neutral pH containing mixed detergents. This procedure yielded a 2,500-fold purification of the medium T antigen in a single step. The protein kinase activity associated with the medium T antigen was also released and was studied in this active state in solution. Sedimentation analysis showed that the bulk of the purified medium T antigen was in a monomeric form (Mr about 42,000) not associated with protein kinase activity. A small fraction of the medium T antigen was found in a rapidly sedimenting form (Mr about 200,000) that possessed protein kinase activity. PMID- 6287462 TI - Bovine papillomavirus vector that propagates as a plasmid in both mouse and bacterial cells. AB - We report the construction of a bovine papillomavirus (BPV)-derived recombinant plasmid that propagates as an extrachromosomal element in both mouse and bacterial cells. Plasmids composed of a subgenomic transforming fragment of BPV DNA, a deletion derivative of pBR322, and a 7.6-kilobase fragment of DNA from the human beta-globin gene cluster efficiently induce focus formation on mouse C127 cells. BPV-beta-globin hybrids are maintained in the transformed cells as plasmids with a copy number of about 10-30 per cell. Plasmids indistinguishable from the input DNA have been recovered by transformation of bacteria with low molecular weight DNA from transformed mouse cells. The human beta-globin gene linked to BPV DNA is transcribed from its own promoter at a high level in these cells. The expression of BPV-linked cellular genes in conjunction with the ability to shuttle DNA between bacteria and mammalian cells may provide a rapid means of analyzing and recovering genes that confer an identifiable phenotype upon mammalian cells. PMID- 6287463 TI - Soybean leghemoglobin gene family: normal, pseudo, and truncated genes. AB - Leghemoglobin (Lb) genes in soybean represent a small family of closely related genes. Three Lb sequences isolated from a genomic library were analyzed at the nucleotide sequence level. A Lb gene present on an 11.5-kilobase (kb) EcoRI genomic fragment spans approximately 1,200 nucleotides and is interrupted at amino acid positions 32 to 33, 68 to 69, and 103 to 104. The intervening sequences, as well as the 5' and 3' flanking regions of this gene, contain the consensus sequences found in other eukaryotic genes. The length of the 5' untranslated region is 49 bases as determined by nuclease S1 mapping. R-loop analysis of the DNA from the recombinant phage containing the 11.5-kb EcoRI genomic fragment showed that another Lb gene is located 2.5 kb away. The nucleotide sequence of the second gene showed that this gene is incomplete, containing only exons 3 and 4. The deduced amino acid sequence of this gene, although showing 76% homology with the corresponding region of the other Lb gene, is not represented in any of the known Lb proteins. Both genes are oriented in the same direction with respect to the coding strand. Analysis of the sequence present on a second genomic clone containing a 4.2-kb EcoRI fragment revealed a truncated Lb gene showing homology with the last exon and the noncoding region at the 3' end of the two other Lb genes. PMID- 6287464 TI - Human DNA sequence homologous to the transforming gene (mos) of Moloney murine sarcoma virus. AB - We describe the molecular cloning of a 9-kilo-base-pair BamHI fragment from human placental DNA containing a sequence homologous to the transforming gene (v-mos) of Moloney murine sarcoma virus. The DNA sequence of the homologous region of human DNA (termed humos) was resolved and compared to that of the mouse cellular homolog of v-mos (termed mumos) [Van Beveren, C., van Straaten, F., Galleshaw, J.A. & Verma, I.M. (1981) Cell 27, 97-108]. The humos gene contained an open reading frame of 346 codons that was aligned with the equivalent mumos DNA sequence by the introduction of two gaps of 15 and 3 bases into the mumos DNA and a single gap of 9 bases into the humos DNA. The aligned coding sequences were 77% homologous and terminated at equivalent opal codons. The humos open reading frame initiated at an ATG found internally in the mumos coding sequence. The polypeptides predicted from the DNA sequence to be encoded by humos and mumos also were found to be extensively homologous, and 253 of 337 amino acids were shared between the two polypeptides. The first five NH2-terminal and last two COOH-terminal amino acids of the humos gene product were in common with those of mumos. In addition, near the middle of the polypeptide chains, four regions ranging from 19 to 26 consecutive amino acids were conserved. However, we have not been able to transform mouse cells with transfected humos DNA fragments or with hybrid DNA recombinants containing humos and retroviral long terminal repeat (LTR) sequences. PMID- 6287465 TI - Coding sequence for the pT181 repC product: a plasmid-coded protein uniquely required for replication. AB - pT181 is a 4.4-kilobase plasmid from Staphylococcus aureus specifying tetracycline resistance and present in about 20 copies per cell. The existence of a diffusible pT181 product required for plasmid replication has been proposed on the basis of trans-complementable thermosensitive mutants defective in plasmid maintenance (phenotype Tsr). In this report, the Tsr mutants are shown to have primary replication defects, and the genetic complementation data are confirmed biochemically. All of five mutations are in a single cistron, the repC cistron; interruption of the plasmid DNA molecule at any of three neighboring restriction sites inactivates repC function. Analysis of the DNA sequence in this region reveals an open reading frame of 939 base pairs which encodes the repC product, a 313-amino acid protein. pT181 replication has been demonstrated in cell-free extracts to require specifically a pT181-coded protein of approximately the same size, and it is proposed that this protein is, indeed, the repC product. Preliminary evidence is discussed suggesting that the pT181 replication rate is controlled at the level of synthesis of the repC protein. PMID- 6287466 TI - Novel class of mouse mammary tumor virus-related DNA sequences found in all species of Mus, including mice lacking the virus proviral genome. AB - Mice in breeding colonies of feral Mus musculus brevirostris (Azrou, Morocco), M. m. musculus (Studenec, Czechoslovakia), and M. m. molossinus (Fukuoka, Japan) were found to lack the mouse mammary tumor virus (MMTV-alpha) proviral genome in their germ line. MMTV-alpha proviral genomes have been found in all inbred strains of M. musculus by using high-stringency nucleic acid hybridization conditions. We conclude that feral mice in these colonies are heterozygous for a limited number of MMTV-alpha proviral genomes and that those lacking them arose as a result of random chromosomal segregation. All mice in another breeding colony of feral M. m. musculus (Sladeckovce, Czechoslovakia) lack MMTV proviral genes. By relaxing the conditions of nucleic acid hybridization, MMTV-related sequences (designated MMTV-beta) were detected in restricted cellular DNA from MMTV-negative mice and all other inbred strains and feral species of the genus Mus. The apparent ubiquity of the MMTV-beta DNA sequences in the genus Mus and the lack of variation in the pattern of restriction fragments containing these sequences within a species distinguishes them from MMTV-alpha. These results suggest that the MMTV-beta DNA sequences either are the evolutionary progenitors of the infectious MMTV genome or represent an accumulation of evolutionarily divergent MMTV-alpha insertions into the mouse germ line. PMID- 6287467 TI - Joining of immunoglobulin heavy chain gene segments: implications from a chromosome with evidence of three D-JH fusions. AB - A chromosomal segment with a unique structure around the immunoglobulin heavy chain joining region (JH) has been molecularly cloned from an Abelson murine leukemia virus-transformed cell line. Attached to JH3 in the cloned DNA, in inverted sequence, is the DNA from JH1 to the JH2 recognition sequence. The inverted segment is attached at its other end to the 5' recognition sequence of a diversity segment (D). To form this structure, three joining events must have occurred on the same chromosome. One of these events could have been a normal D JH joining but the others must have been irregular events including ones that result in inversions. One of the joining events left fused recognition elements from JH2 and a D whose sequence shows that, during joining, reciprocal joinings of the recognition elements must occur to fuse the heptameric elements back to back. Because joined D and JH undergo deletion of terminal coding sequence during recombination but the joined heptameric recognition sequences do not contain the deleted sequence, joining must be a nonreciprocal event. Also, extra nucleotides are inserted between D and JH as part of the joining process; it is suggested that this added sequence is a product of the activity of terminal deoxynucleotidyltransferase at the D/JH (and probably the VH/D) joints and that it represents a new element of heavy chain gene structure, the N region. PMID- 6287468 TI - sigma, a repetitive element found adjacent to tRNA genes of yeast. AB - sigma is a DNA element of about 340 base pairs (bp) that is repeated many times in the yeast genome. The element has 8-bp inverted repeats at its ends and is flanked by 5-bp direct repeats. The 5-bp repeats are different for each sigma and have no homology with the ends of the sigma sequence. sigma is located 16 or 18 bp from the 5' end of several tRNA genes. Southern analysis of different yeast strains shows that the pattern of hybridization is different even for closely related strains. PMID- 6287469 TI - Insertion of a movable genetic element, 297, into the T-A-T-A box for the H3 histone gene in Drosophila melanogaster. AB - From a cloned library of Drosophila DNA, a recombinant phage was isolated which contained a nucleotide sequence homologous to both the histone gene and a movable genetic element, 297 [Saigo, K., Millstein, L. & Thomas, C.A., Jr. (1981) Cold Spring Harbor Symp. Quant. Biol. 45, 815-827]. We have determined the nucleotide sequences of the termini of 297 as well as its insertion site in the histone gene in the clone. Our results strongly suggest that (i) although 297 ends with typical long terminal repeats, integrated 297 is not bounded by 5' T-G...C-A 3', dinucleotides commonly found at the very ends of integrated forms of other similar genetic elements, and (ii) the insertion of 297 occurred exactly within the T-A-T-A box for the H3 histone gene containing T-A-T-A-T-A. The structures of the insertion sites of 297 were also examined by using two other recombinant clones of the histone gene. A 6-base-pair-long segment, T-A-T-A-T-A, was again found to be present at the insertion sites of 297 in both clones. Based on these results, it was concluded that, at least within the histone gene cluster, the insertion of 297 was carried out in a site-specific fashion with T-A-T-A-T-A as the target sequence. PMID- 6287470 TI - Regulation of the in vitro antibody response by neuroendocrine hormones. AB - Treatment of lymphocytes with inducers of interferon alpha (IFN-alpha) results in the production of corticotropin (ACTH) and endorphin-like activities. The pro opiomelanocortin-derived hormones ACTH and alpha-, beta-, and gamma-endorphin and the structurally related hormones [Leu]- and [Met]enkephalin were therefore tested for their effects on the in vitro antibody response of mouse spleen cells. ACTH and alpha-endorphin were potent inhibitors (>/=80% suppression) of the antibody response to the T-cell-dependent antigen sheep erythrocytes at a concentration of 0.5 muM. [Met]- and [Leu]enkephalin were moderate inhibitors (approximately 60% suppression) at 0.2-2 muM, and beta- and gamma-endorphin were minimal inhibitors (approximately 20% suppression) at 5-6 muM. At higher concentrations ACTH also inhibited the antibody response to the T-cell independent antigen dinitrophenyl-Ficoll, suggesting that T-cell function was more sensitive to blockage by these hormones than was B-cell function. ACTH and IFN had similar suppression properties; thus, the hormone-like activities associated with IFN-alpha may play a role in IFN-induced immunosuppression. alpha Endorphin immunosuppression was blocked by naloxone, which suggested that alpha endorphin exerted its effects through binding to opiate-like receptors on the spleen cells. The failure of beta-endorphin to suppress the immune response significantly was not due to its failure to bind to the opiate-like receptors because it blocked alpha-endorphin-induced suppression. Direct evidence for both opiate and ACTH receptors on the spleen cells was obtained in binding studies with labeled enkephalin and ACTH. Such studies revealed the presence of both high and low-affinity receptors. The data show that neuroendocrine polypeptide hormones can regulate the immune response. PMID- 6287471 TI - Non-immunoglobulin-associated DNA rearrangements in mouse plasmacytomas. AB - We have characterized a class of DNA rearrangements in plasmacytomas. These recombination events involve a DNA sequence whose origin is outside of the locus of the heavy chain constant region genes, CH. Therefore, we choose to refer to this sequence as non-immunoglobulin-associated rearranging DNA (NIARD). We have isolated two abortively rearranged C alpha genes, generated by NIARD events from the alpha-producing J558 myeloma. Restriction endonuclease maps of these sequences reveal two possible recombination sites in NIARD that are separated by approximately 6.5 kilobase pairs of DNA (defined as 5' and 3' sites). A NIARD rearrangement occurs in 15 out of 20 plasmacytomas tested, including gamma 3-, gamma 1-, gamma 2b-, gamma 2a-, and alpha-producers, but this event usually does not involve a CH switch (S) region. In fact, only S alpha appears to accept NIARD. However, NIARD did not undergo a rearrangement in eight IgA-producing hybridomas tested. One germ-line copy of NIARD (a 22-kilobase pair EcoRI fragment) is retained in all plasmacytomas. NIARD does not appear to possess repetitive DNA sequences homologous to S mu or S alpha. We discuss the possible role and implications of NIARD-like sequence rearrangements in allelic exclusion and chromosomal translocation events in plasmacytomas. PMID- 6287472 TI - Effects of electromagnetic stimuli on bone and bone cells in vitro: inhibition of responses to parathyroid hormone by low-energy low-frequency fields. AB - Low-energy electromagnetic fields pulsed at frequencies of 10-90 Hz significantly increase healing of chronic fracture nonunions in man. These fields are effective at tissue current levels several orders of magnitude lower than those required for transmembrane depolarization of normal cells. We have examined the effects of two clinically used pulsed electromagnetic fields on cultures of the osteoblast like mouse bone cell line MMB-1. Both fields significantly reduced cellular production of cAMP in response to parathyroid hormone and osteoclast activating factor. Neither basal nor fluoride-activated levels of adenylate cyclase were altered in membranes from cells cultured in the fields; however, the same membrane preparations exhibited markedly inhibited responses to parathyroid hormone. The fields blocked the inhibitory effects of the hormone on collagen synthesis by MMB-1 cells. However, there was no effect on the inhibition of collagen synthesis by 1,25-dihydroxyvitamin D(3), which is believed to act primarily by a nuclear, rather than by a membrane-dependent, mechanism. No significant differences were noted between effects of the two fields, one generating continuous pulse trains (72 Hz) and the other generating recurrent bursts (15 Hz) of shorter pulses. We hypothesize that these field effects are mediated primarily at the plasma membrane of osteoblasts, either by interference with hormone-receptor interactions or by blocking of receptor-cyclase coupling in the membrane. These responses occurred with induced extracellular fields of 1 mV/cm or less, even though transmembrane potential gradients are typically 10(5) V/cm. PMID- 6287473 TI - Platelets of pseudohypoparathyroid patients: evidence that distinct receptor cyclase coupling proteins mediate stimulation and inhibition of adenylate cyclase. AB - We studied platelets of patients with the genetic disorder pseudohypoparathyroidism (PHP) to test whether the nucleotide-binding proteins mediating stimulation of adenylate cyclase (termed N(s)) are identical to those mediating inhibition of cyclase (termed N(i)). Functional responses to hormones that work through stimulation of adenylate cyclase are blunted in PHP patients. The erythrocytes of many of these patients (PHP-Ia) have previously been shown to have decreased N(s) activity whereas those of other PHP patients (PHP-Ib) have normal N(s) activity. We find that this decreased N(s) activity (measured by the ability to restore adenylate cyclase activity to membranes prepared from S49 cyc( ) cells) also occurs in the platelets of PHP-Ia but not of PHP-Ib patients. Platelets from both groups of patients accumulate less cAMP in response to prostacyclin than do platelets from control subjects. In contrast to the decreased N(s) function in patients with PHP-Ia, we find that N(i) function in platelets is similar in these patients and control subjects in several types of experiments: (i) epinephrine-mediated inhibition of prostacyclin-stimulated cAMP production in intact platelets; (ii) the affinity of platelet alpha(2)-adrenergic receptors for epinephrine, as determined by competition for [(3)H]yohimbine binding; (iii) the decrease in receptor affinity for epinephrine produced by Na(+) and GTP; and (iv) the concentration dependence of GTP for decreasing the affinity of these receptors for epinephrine. Because N(i) is expressed normally in platelets from patients that are genetically deficient in N(s), we conclude that N(s) and N(i) are likely to be distinct gene products. PMID- 6287474 TI - Ca2+ -activated K+ conductance in internally perfused snail neurons is enhanced by protein phosphorylation. AB - Depolarizing voltage steps induce inward and outward currents in voltage-clamped, internally perfused neurons from the snail Helix roseneri. Addition of the catalytic subunit of cyclic AMP-dependent protein kinase (ATP:protein phosphotransferase, EC 2.7.1.37) to the internal perfusing medium results in an increase in the net outward current, with no apparent effect on the inward current. Catalytic subunit inactivated by 5,5'-dithiobis(2-nitrobenzoic acid) is without effect, indicating that the increase in net outward current results from protein phosphorylation rather than an unspecific effect of protein perfusion. Decreasing the external Ca2+ concentration from 10 to 1 mM eliminates the effect of catalytic subunit, suggesting that Ca2+ plays an important role in this response. This suggestion is supported by the fact that the stimulation by catalytic subunit can be mimicked by increasing the Ca2+ concentration in the internal perfusion medium and can be prevented by intracellular perfusion with 10 mM EGTA. The results are consistent with the hypothesis that cyclic AMP-dependent protein phosphorylation regulates the Ca2+-activated K+ conductance in these cells. PMID- 6287476 TI - Ethanol disordering of spin-labeled mouse brain membranes: correlation with genetically determined ethanol sensitivity of mice. AB - Disordering of brain and erythrocyte membranes by ethanol in vitro was measured by ESR using 5-doxylstearic acid as spin label. Synaptosomal plasma membranes and erythrocyte membranes were isolated from two lines of mice developed, by selective breeding, for differential sensitivity to hypnotic effects of ethanol. Membranes taken from alcohol-sensitive "long-sleep" mice were more strongly disordered by ethanol in vitro than were membranes from alcohol-resistant "short sleep" mice. Furthermore, within a population of genetically heterogeneous mice, the most ethanol-sensitive animals had the most ethanol-sensitive synaptosomal plasma membranes. In vivo sensitivity of the individual mice was evaluated by measuring brain ethanol levels at a precise behavioral end point, recovery from ataxia. The data extend our previous observations of correlations between in vitro and in vivo effects of ethanol and suggest that membrane disordering may be a primary mechanism of acute effects of ethanol. PMID- 6287477 TI - Effect of cholecystokinin octapeptide on food intake in pigs. PMID- 6287475 TI - beta-Endorphin enhances lymphocyte proliferative responses. AB - The opioid peptides alpha- and beta-endorphin and [D-Ala2, Met5]enkephalin were investigated for their effect on the proliferation of resting and activated rat splenic lymphocytes in vitro. beta-Endorphin enhanced the proliferative response of spleen cells to the T cell mitogens concanavalin A and phytohemagglutinin. The effect of beta-endorphin was dose dependent and occurred at peptide concentrations similar to those found in rat plasma. alpha-Endorphin and [D-Ala2, Met5]enkephalin did not affect the proliferative responses to any mitogen tested. Furthermore, the potentiating effect of beta-endorphin was not reversed by treatment with 10 microM naloxone. None of the peptides had any effect on resting, unstimulated spleen cells or on the response to a mixture of lipopolysaccharide and dextran sulfate, which is specifically mitogenic for B lymphocytes. The pharmacological properties of the beta-endorphin potentiation indicate that the effect may be mediated by a nonopiate but beta-endorphin specific mechanism. These results suggest a possible role for peripheral beta endorphin and may provide a link between stress and disease susceptibility. PMID- 6287478 TI - Trace element intake by Asians during pregnancy. PMID- 6287479 TI - Molecular cloning and sequence analysis of highly repetitive DNA sequences contained in the eliminated genome of Ascaris lumbricoides. AB - High molecular weight DNA from germ line and somatic cells of the DNA eliminating nematode Ascaris lumbricoides has has been isolated and digested with different restriction enzymes. The resulting DNA fragments were separated by agarose gel electrophoresis. Germ line but not somatic DNA shows a prominent band about 120 bp long as well as multiples of that length. These fragments are shown to be monomers and multimers of a highly repetitive satellite DNA, which is eliminated mostly but not completely during the process of chromatin diminution. Restriction digests, hybridization experiments and sequence analysis revealed that this eliminated satellite is composed of a whole set of different but related variant classes, all of them showing the same repeating unit length of about 120 bp. Members of the same variant class are tandemly linked and therefore physically separated from other variant classes. All satellite sequences can be derived from the same common ancestor sequence, differing only by base substitutions, insertions and deletions. There is no evidence for transcription of satellite DNA at any stage and tissues analyzed. PMID- 6287480 TI - The conalbumin "TATA" box sequence and the SV40 72 base-pair repeat region influence expression of a chimeric gene in vivo. PMID- 6287481 TI - Characterization and sequence analysis of interspersed repetitive DNA sequences transcribed in X.laevis embryos. PMID- 6287482 TI - DNA mediated transfer and fate of a proviral gene of mouse mammary tumor virus in cultured cells. PMID- 6287483 TI - Opioid regulation of pituitary secretion. PMID- 6287484 TI - The catechol estrogens: physiologic studies. PMID- 6287485 TI - Effects of prolactin on the male gonad and sex accessory organs. PMID- 6287486 TI - Antimetastatic action of RX-RA 69, a new potent PDE-inhibitor in the Lewis lung carcinoma of the mouse. AB - RX-RA 69, a pyrimido-pyrimidine derivative, is a new potent PDA-inhibitor which inhibits tumor cell induced platelet aggregation in vitro. Also ex vivo an inhibition was found after pretreating mice with 1 mg/kg RX-RA 69 orally one hour before collecting the blood. The same dosage is able to prevent the drop in platelet count induced by injecting a large number of tumor cells into mice. The same dosage schedule in the amputation experiment resulted in a reduction of incidence of matastases in all treated groups; the best results, however, were obtained by starting the treatment two days after the amputation. In the spontaneous model an inhibition of metastases formation by 10-20 mg/kg RX-RA 69 starting from the fourth day after tumor cell implantation was found in four independent experiments. The mechanism of the antimetastatic action of this drug remains to be evaluated. PMID- 6287487 TI - A mitogenic factor for transformed cells from human platelets. PMID- 6287488 TI - Inhibition of the platelet-aggregating activity of two human adenocarcinomas of the colon and an anaplastic murine tumor with a specific thrombin inhibitor: dansylarginine N-(3-ethyl-1, 5-pentanediyl)amide. PMID- 6287489 TI - The effects of various tryptolines (tetrahydro-beta-carbolines) on 5 hydroxytryptamine receptors. PMID- 6287490 TI - beta-Carbolines and benzodiazepine receptors. PMID- 6287491 TI - beta-Carbolines and benzodiazepine receptors: structure-activity relationships and pharmacologic activity. PMID- 6287492 TI - Do benzodiazepine receptors play a role in sleep regulation? Studies with the benzodiazepine antagonist, 3-hydroxymethyl-beta-carboline (3-HMC). PMID- 6287493 TI - Neurotransmitter, opiate and benzodiazepine receptor binding of tetrahydroisoquinolines and beta-carbolines in brain membranes. PMID- 6287495 TI - Is there a correlation between the concentration of beta-carbolines and their pharmacodynamic effects? AB - Two sensitive and specific methods are presented for the determination of 6-OH tetrahydronorharmane (6-OH-THN, 6-OH-THBC) and harmane (1-Me-BC). The concentration of 6-OH-THN in blood platelets from men was found 5.19 +/- 0.57 ng (mean +/- SEM) per 10(9) platelets, of acute schizophrenic patients 2.66 +/- 0.38 ng per 10(9) platelets, p less than 0.02, and in rats 6 - 13 ng x 10(9) platelets. The concentration of harmane in the urine of rats was measured 9.72 +/ 1.56 ng per 16 h. A load with ethanol caused an increased excretion of the beta carboline in some rats. In pharmacological experiments substantial evidence was detected for a correlation between the (3H)-flunitrazepam displacing potency and the conflict-augmenting effects of beta-carbolines. Furthermore, a good correlation was found between the results of binding experiments and the antagonism of the beta-carbolines with respect to the activating effect of low doses of diazepam. No such correlation exists for the sedative effect. PMID- 6287494 TI - A convergent approach to the pharmacology of tetrahydroisoquinolines. PMID- 6287496 TI - Germ-cell tumors of the diencephalon. AB - Germ-cell tumors arising in the neuraxis have similar histopathological characteristics to those germ-cell tumors occurring in the gonads and other midline body sites. As a rare cause of intracranial neoplasm, germ-cell tumors affect predominantly the diencephalon and in particular the pineal region and suprasellar structures. Clinical features are due to local infiltration of these areas by tumor and hydrocephalus. Germinomas or mixed-cell tumors with a germinoma component make up the majority of these tumors and are very responsive to irradiation and chemotherapy. Early clinical recognition of a diencephalic tumor and determination of the specific tumor-cell type may result in curative treatment by surgery, irradiation and chemotherapy. PMID- 6287497 TI - Melatonin and corticosteroid response to metyrapone in patients with pituitary disease. AB - Melatonin secretion is dependent both on the environmental lighting and noradrenergic control and seems to be related to the ACTH-cortisol secretion. The aim of the present study was to further elucidate the relationship between ACTH cortisol and melatonin. For this purpose we chose the Metyrapone test. The melatonin secretion was studied in 10 patients, and increased significantly in 8/10 during and in one after the Metyrapone administration. These results further support a relationship between melatonin and ACTH-cortisol secretion, possibly through a suprapituitary interaction. PMID- 6287498 TI - Probing the chick pineal clock. PMID- 6287499 TI - Interaction between pargyline, a monoamine oxidase inhibitor, and beta-adrenergic receptors in the rat pineal gland. AB - The hypothesis that pargyline, a monoamine oxidase inhibitor, may interact with pineal beta-adrenergic receptors to increase N-acetyltransferase (NAT) activity and, thereby, melatonin content was tested in intact and superior cervical ganglionectomized (SCGX) rats some of which were also treated with the tryptophan hydroxylase inhibitor para-chlorophenylalanine (PCPA). Pargyline injection increased both NAT activity and melatonin content, an effect nullified by conjunct propranolol injection. While pargyline injection also increased NAT activity in intact, PCPA-treated rats, the effects of this drug were most evident in SCGX, PCPA-treated rats. PCPA treatment prevented pargyline-induced increases in melatonin content in intact and SCGX rats suggesting an artificially-induced dissociation of melatonin biosynthesis from its rate-limiting enzyme. We suggest that pargyline may be capable of interacting with pineal beta-adrenergic receptors to increase NAT activity in the absence of pineal norepinephrine (NE) and serotonin. PMID- 6287500 TI - Dietary requirements and functions of alpha-linolenic acid in animals. PMID- 6287501 TI - Protection against ethanol-induced embryonic damage by administering gamma linolenic and linoleic acids. AB - Many reports have now confirmed the teratogenic potential of alcohol in humans and in laboratory animals. A characteristic pattern of congenital anomalies is present in infants born to mothers suffering from chronic alcoholism. The pathogenesis of this condition is unclear. Chronic consumption of ethanol causes a depletion of essential fatty acids, partly by blocking gamma-linolenic acid formation and partly by depleting dihomogammalinolenic acid. Whether this action of ethanol on essential fatty acid and prostaglandin metabolism may account for its teratogenic potential was investigated in the rat. Treatment of pregnant rats with ethanol and evening primrose oil (efamol), a rich source of gammalinolenic acid, led to a significant reduction in the embryopathic activity of ethanol. PMID- 6287502 TI - Effect of methaqualone on plasma corticosterone in rats: possible sites of action. AB - Methaqualone produces large increases in plasma corticosterone in rats. This effect does not involve a direct stimulation of either the adrenal glands or pituitary, nor is it the result of a significant alteration in the clearance of circulating corticosterone. Methaqualone may therefore be influencing brain mechanisms which ultimately regulate hypothalamic CRH secretion, although a peripheral site of action has not yet been ruled out. PMID- 6287503 TI - Extreme hyperthermia induced in cats by the enkephalin analog FK 33-824. AB - FK 33-824 [H2N-Tyr-D-Ala-Gly-MePhe-Met(O)-OH] was injected into the third cerebral ventricle of unrestrained cats. Doses of 0.25-4 microgram induced dose related increases in body temperature. Hyperthermic responses to 1 microgram of the peptide were greater the warmer the environment. Naloxone given intraventricularly 1 hr after FK 33-824 (1 microgram) reduced the hyperthermia. In 12 tests with six cats FK 33-824 (1-25 microgram) increased temperature 4.2 4.7 degrees C. These marked responses were also inhibited by naloxone, but two cats died when administration of antagonist was delayed for 80 min to 3 hr after attainment of maximal body temperature. Larger doses of FK 33-824 (50-250 microgram) evoked little increase in temperature, indicative of a separate action to depress thermoregulation. Although responses to FK 33-824 were antagonized by naloxone, this peptide, like another enkephalin analog D-Ala2-Met enkephalinamide, must act on receptors which are not affected by morphine since (1) the hyperthermic response to FK 33-824 varied with environmental temperature, whereas the response to morphine does not, and (2) high doses of FK 33-824 depressed thermoregulation, an activity not shared by morphine in the cat. Furthermore, the maximal increases in temperature after FK 33-824 injection were greater than those evoked by either morphine or D-Ala2-Met-enkephalinamide. This observation provides evidence for an additional subset of naloxone-sensitive, v, receptors, stimulation of which can influence thermoregulation in the cat. PMID- 6287504 TI - [Stereoisomers of a substituted 1-benzopyrano [4,3-b]-1,5-benzodiazepine as inhibitors of cAMP-phosphodiesterase]. PMID- 6287505 TI - Metabolism of polycyclic aromatic hydrocarbons: etiologic role in carcinogenesis. PMID- 6287506 TI - In vivo identification and distribution of alpha- and beta-adrenoceptors in rat heart and lung. AB - Uptake of 3H-prazosin (3H-PZ), a potent alpha-adrenoceptor antagonist, and 3h dihydroalprenolol (3H-DHA), a potent beta-antagonist, was measured in rat heart and lung after intravenous injection. 3H-PZ binding was inhibited in a dose dependent manner by phentolamine with maximum displacement at 5 mg/kg which represented 50-70% of the total binding. 3H-DHA was inhibited by increasing doses of l-propranolol with maximum displacement at 1 mg/kg (30-70% of total binding), whereas d-propranolol was approximately 100 times less potent, confirming stereospecificity of binding. These radioligands appeared to label adrenoceptors in vivo. The distribution of both radioligands in heart, lung and blood vessels was investigated. PMID- 6287507 TI - Photoinactivation of enzymes by linear and angular furocoumarins. PMID- 6287508 TI - Pressure variation of enzymatic reaction rates: IV. Xanthine oxidase and superoxide dismutase. AB - Effect of pressure on activities of O-(2)-producing enzyme xanthine oxidase (XO) and O-(2)-scavenging enzyme superoxide dismutase (SOD) has been investigated to 1000 bar. Methods used included spectrophotometric determinations of inhibition by SOD of O-(2)-induced reduction of cytochrome c and oxidation of ascorbic acids. It was found that (1) pressure increases XO activity, the activation volume being delta V not equal to = -45 +/- 5 cm3/mol; and (2) pressure decreases SOD activity, the activation volume being delta V not equal to = 30 +/- 5 cm3/mol. Under the conditions of these experiments, pressure therefore favors an increase in O-(2) activity. This may lead to increased cell damage in organisms in high-pressure situations even if oxygen concentration is held constant. PMID- 6287509 TI - Interaction between synthetic lecithin and various sulfur-containing radioprotectors. AB - Interaction of the synthetic lecithin dipalmitoylphosphatidylcholine (DPPC) smectic mesophases with sulfur-containing radioprotectors was investigated by means of spectrophotometric and dielectric measurements. Electrical conductivity behavior indicated in all cases that an electrostatic interaction occurs between DPPC and the antiradiation drugs. This interaction is very strong in the case of the WR 2721 molecule. Thermal transitions of DPPC studied by spectrophotometry and conductivity Arrhenius diagrams showed that although the radio-protectors investigated (except WR 2721) delete the pretransition of the lipid phase, the principal transition is not modified. The observed electrostatic interactions are discussed with regard to ionized sites of the phosphatidylcholine lipid head and those of the radioprotectors. The special cases of WR 2721 and its metabolite, WR 1065, are examined. PMID- 6287510 TI - Effects of various coumarins from roots of Angelica dahurica on actions of adrenaline, ACTH and insulin in fat cells. PMID- 6287511 TI - Limitations of the sternocleidomastoid musculocutaneous flap in head and neck cancer reconstruction. PMID- 6287512 TI - The revascularization of pedicle skin flaps in pigs: a functional and morphologic study. AB - Functional and morphologic changes occurring during the revascularization of pedicle flaps have been investigated in the skin of pigs. The skin flaps, 16 cm long by 4 cm wide, were based on a row of segmental vessels arising from the internal mammary artery. Comparative measurements were made in flapped and normal skin. The inherent blood supply in the pedicle of the flap was unable to maintain the whole of the flap in a viable state. Flap viability was ascertained at surgery by the use of the intravital dye Disulphine blue. Injections of the dye after surgery gave a less accurate prediction of viability than when dye was injected prior to surgery. Revascularization between the flap and surrounding skin was evident 3 to 4 days postoperatively at the distal, most hypoxic part of the viable flap. The whole flap had a collateral vascular supply 7 to 10 days after surgery. Isotope clearance studies showed that the greatest functional changes occurred in the distal third of the viable flap, where, after initially slowing, the clearance rate became faster than in normal skin (day 5). Potassium extraction studies indicated similar changes. However, an increase in the red cell volume on day 1 suggested that vascular shunting was occurring. The results of the morphologic studies indicated a correlation between the number of blood vessels per unit area, the thickness of the dermis, and the recorded functional changes. Seven days after surgery, when isotope clearance rates were very rapid, there was a significant increase in the vascular density and dermal thickness. PMID- 6287513 TI - Possible subsensitization of alpha 2-adrenergic receptors by chronic monoamine oxidase inhibitor treatment in psychiatric patients. AB - Clonidine was administered to nine psychiatric patients before and after chronic treatment (3 to 4 weeks) with clorgyline, a selective monoamine oxidase type A inhibitor with antidepressant efficacy. The hypotensive response to clonidine, believed to be mediated by brain alpha 2-adrenergic receptors, was significantly attenuated by chronic but not acute (2 to 3 days) clorgyline treatment, with a time course similar to the onset of its clinical efficacy. This study supports the hypothesis that subsensitization of alpha 2-adrenergic receptors plays an important role in clorgyline's antidepressant effects and may constitute a key contribution to the mode of action of other antidepressant treatments as well. PMID- 6287514 TI - Echovirus 11 infections of newborns with mortality during the 1979 enterovirus season in Milwaukee, Wis. AB - Echovirus serotype 11 (ECHO-11) was implicated in three neonatal deaths during an enterovirus outbreak from July through October 1979 in Milwaukee. The deaths followed congenital infections acquired in the community during late pregnancy. Two of the three ECHO-11 and one Coxsackie B4 deaths of infants occurred after cesarean section deliveries. Of 225 confirmed echovirus infections, 30 to 45 percent occurred in infants under 60 days old, 54 to 67 percent in the first year of life, and 13 to 25 percent in the over-10 age groups. In 13 cases with onset of symptoms in the first week of life. 8 (including the 4 fatalities) were acquired congenitally; 6 of the 8 were associated with ECHO-11, 2 with ECHO-7, and 1 with Coxsackie B4. ECHO-7 and 30 other predominant strains were isolated during the outbreak, but none was associated with mortality or severe disease in neonates. At a Milwaukee hospital, a temporal association was observed between echovirus infection, particularly ECHO-11, and increased numbers of stillbirths. PMID- 6287515 TI - The natriuretic hormone. PMID- 6287516 TI - The effects of black widow spider venom on the innervation of muscles paralysed by botulinum toxin. AB - Botulinum toxin (BoTx) was injected into the muscles of one leg in mice causing local paralysis. Black widow spider venom (b.w.s.v.) was then injected into the paralysed muscles 3 or 15 d later. In both groups b.w.s.v. destroyed the nerve terminals poisoned by BoTx. In the 15 d group axonal sprouts, which had former due to the block of neuromuscular transmission by BoTx, were also destroyed. Within a few days the motor nerve terminal regenerated and the muscles recovered from paralysis at a faster rate than after BoTx alone. Recovery seemed to begin earlier in muscles where axonal sprouting was already advanced when b.w.s.v. was injected. The normal pattern of innervation was re-established in both groups, which was in marked contrast with muscles after BoTx alone where numerous sprouts and many ectopic end-plates had formed. PMID- 6287517 TI - Ultrasonic examination of the abdomen in malignant disease: Part 2 Other abdominal organs, including kidney, stomach and pancreas. PMID- 6287518 TI - Diagnostic approaches to liver and spleen metastases. PMID- 6287519 TI - Transient early computed tomographic changes mimicking tumor progression after brain tumor irradiation. AB - Transient computed tomographic (CT) changes occurring within three months after radiotherapy are described in three patients with cerebral gliomas. These changes consist of enlargement of an area of central necrosis, new tumor enhancement, and increased tumor enhancement with adjacent low-density changes. Subsequent scans showed regression of these changes in all patients. It is proposed that these changes are the direct effect of radiation on tumor tissue and adjacent normal brain; possible mechanisms are discussed. These radiation-induced changes mimic tumor progression on CT. The importance of recognizing their transient nature is stressed. PMID- 6287520 TI - Transient hepatic attenuation difference of lobar or segmental distribution detected by dynamic computed tomography. PMID- 6287521 TI - Gallium 67 citrate scanning and serum angiotensin converting enzyme levels in sarcoidosis. AB - Gallium 67 citrate scans and serum angiotensin converting enzyme (ACE) levels were obtained in 54 patients with sarcoidosis and analyzed in relation to clinical manifestations. 67Ga scans were abnormal in 97% of patients with clinically active disease (n = 30) and in 71% of patients with inactive disease (n = 24). Serum ACE levels were abnormally high (2 standard deviations above the control mean) in 73% of patients with clinically active disease and in 54% of patients with inactive disease. Serum ACE levels correlated significantly with 67Ga uptake score (r =.436; p less than .005). The frequency of abnormal 67Ga scans and elevated serum ACE levels suggests that inflammatory activity with little or no clinical expression is common in sarcoidosis. Abnormal 67Ga scans were highly sensitive (97%) but had poor specificity (29%) to clinical disease activity. The accuracy of negative prediction of clinical activity by normal scans (87%) was better than the accuracy of positive prediction of clinical activity by abnormal scans (63%). 67Ga scans can be used to support the clinical identification of inactive sarcoidosis. PMID- 6287522 TI - [A new plaque assay for the quantification of Junin virus]. PMID- 6287523 TI - [Effect of experimental infection of pregnant guinea pigs with Junin virus]. PMID- 6287525 TI - Current concepts of bile formation. PMID- 6287524 TI - Acute hemodynamic responses to vasodilator therapy in congestive heart failure. PMID- 6287526 TI - Lymphocyte-instructed monocyte induction of the coagulation pathways parallels the induction of hepatitis by the murine hepatitis virus. PMID- 6287527 TI - Hepatocellular carcinoma: changing concepts in recent years. PMID- 6287528 TI - Glycoproteins on hepatocytic surfaces. PMID- 6287529 TI - Discriminative stimulus properties of the vasodilator, hydralazine: differential generalization with alpha 1 and alpha 2 adrenoreceptor drugs. AB - 1. Male albino rats were trained to an operant procedure of lever pressing on an FR-10 schedule of food reinforcement to respond on one lever located on one side of the food cup after an injection of hydralazine (1,25 mg/kg), and to respond on an alternate lever located on the other side of the food cup after an injection of saline. 2. Seven out of ten rats learned the hydralazine-saline discrimination to the rigid criterion of selecting the correct lever for reinforcement on ten consecutive sessions. 3. The elicitation of the discriminative stimulus was dose dependent (r = 0,98; p less than .001) with 100, 43, and 14% of the subjects selecting the hydralazine lever following hydralazine doses of 1,25; 0,32 and 0,08 mg/kg, respectively (ED50, 0,28 mg/kg). 4. A reduction in response rate and blood pressure was noted only at the 1.25 mg/kg dose. 5. No tolerance to the hypotensive effect of hydralazine was found. 6. In generalization tests, prazosin, an alpha 1 antagonist, was found to produce a dose-dependent generalization to hydralazine (ED50, 1, 25 mg/kg) while clonidine, an alpha 2 agonist, did not generalize. 7. These data indicate that hydralazine produces a discriminable interoceptive stimulus exact site of action of which is not known. PMID- 6287530 TI - Alterations of synaptic high and low affinity opiate binding sites after acute and chronic morphine administration in mice. AB - 1. The opiate receptor binding to whole brain synaptic membranes obtained from mice subjected to acute and chronic morphine administration and precipitated withdrawal was investigated. 2. The number of high and low affinity binding sites was significantly higher in morphine tolerant group than in control; acute administration of morphine also induced an increase in the number of binding sites, though this increase was significantly lower as compared to tolerant group. 3. The affinity of both high and low affinity sites, however remained unchanged after acute or chronic treatment. In contrast, both the affinity and the number of binding sites were significantly reduced after precipitated withdrawal, compared to the tolerant group. 4. Sodium chloride enhanced the antagonist binding and inhibited the agonist binding in both tolerant and non tolerant groups. 5. It is concluded that (a) the increase in the number of receptors during tolerance development is an extension of the acute effect of morphine, (b) the character of tolerant membrane is qualitatively same as that of non-tolerant membrane, (c) upon withdrawal, the receptor population is brought back to normal; the altered higher, affinity after withdrawal may be a compensatory effect as a corollary to the withdrawal-induced decrease in receptor population. PMID- 6287531 TI - Evidence that ethanol-induced impairment of roto-rod performance is not mediated by opioid mechanisms. AB - 1. The performance of mice, trained to remain on a roto-rod, was significantly impaired by both morphine and ethanol. The behavioral impairment was dose dependent. 2. Naloxone (10, 30 and 100 mg/kg) was found to antagonize only the morphine-induced impairment of roto-rod performance. 3. Animals made tolerant to morphine did not perform better than non-tolerant animals when challenged with ethanol, but did perform better when challenged with morphine. As such, no cross tolerance developed between ethanol and morphine in this model. 4. These results suggest that the ethanol-induced impairment of roto-rod performance is not mediated via opioid mechanisms. PMID- 6287532 TI - Prostaglandins and schizophrenia: a review. AB - 1. Clinical observations and experimental findings are rendered which have raised the possibility that schizophrenia may be related to PGE deficiency and/or excess. 2. In favor of PGE1 deficiency are the findings that ADP-induced increase in synthesis and PGE1 stimulated cAMP accumulation are significantly lower in platelets of schizophrenic patients than in normal controls. 3. In favor of PGE excess are the findings that the concentration of immunoreactive PGE in cerebrospinal fluid of schizophrenics is higher than that of healthy controls, neurotic patients and patients undergoing neurological examination. 4. An argument against the PG excess hypothesis is that paracetamol, a substance which reduces PG levels has no therapeutic effect in schizophrenia. 5. The two hypotheses--PGE deficiency and PGE excess--are not compatible because of the bell shape dose response curve with PG's in certain biological systems. PMID- 6287533 TI - [Movable genetic elements in bacteria (author's transl)]. PMID- 6287534 TI - [Mechanism of cap formation and its function in protein synthesis initiation (author's transl)]. PMID- 6287535 TI - [Chemical synthesis of proteins (author's transl)]. PMID- 6287536 TI - Prostaglandins: specific inhibition of platelet adhesion to collagen and relationship with cAMP level. AB - The effect of 3 prostaglandins (PG's) (I2, D2 and E1) on the adhesion of platelets to purified type III collagen has been investigated. A quantitative method for a specific evaluation of the adhesion has been applied and has revealed an inhibition of adhesion by low concentrations (10(-10)M) of PGs added before collagen; the effect varied as a function of the dose of PGs (maximum at 10(-6)M) which also induced an increase in the level of platelet cAMP. The inhibition of adhesion and the elevation of platelet cAMP followed the same time course and were either of short duration (rapid decrease in the induced effects after 15 and 45 seconds in the case of PGE1) or longer lasting (maximum effect maintained for 5 minutes in the case of PGI2 and D2). These effects were potentiated by a phosphodiesterase inhibitor such as theophylline (10(-3)M). The addition of PGs after collagen resulted in a reduction of the enhancement of cAMP, associated with a decrease in the inhibition of adhesion. Moreover, the addition of exogenous cAMP (dibutyryl N6-02' cAMP) induced a comparable inhibition. A correlation between the adhesion of platelets to collagen and the level of either endogenous or exogenous cAMP has been established. The PGs also inhibited the platelet release reaction from the alpha granules (beta TG) and the dense bodies. (5-HT and ADP). A greater inhibition of release than of adhesion was observed for the same doses of PGs added. PMID- 6287537 TI - Viruses in water and soil. PMID- 6287538 TI - Corticotrophin-related peptides in the intermediate lobe of the rodent pituitary gland: characterization by high performance liquid chromatography and radioimmunoassay. AB - High performance liquid chromatography (HPLC) and radioimmunoassay have been used to characterize corticotrophin-related peptides in extracts of the intermediate lobe of the rat and mouse pituitary gland. Multiple peaks have been observed, which resemble corticotrophin-like intermediate lobe peptide (CLIP) in that they cross-react with antisera raised against the COOH-terminal region of corticotrophin (ACTH) but not against NH2-terminal directed antisera. These CLIP like peptides were released from the incubated neurointermediate lobe and their secretion was inhibited in the presence of dopamine. Heterogeneity of peptide species was also observed with antisera raised against alpha-MSH. Multiple peaks of CLIP and alpha-MSH-like material were identified in pituitary extracts from the mouse and levels were elevated in the genetically obese (ob/ob) animal. The nature and possible functions of multiple forms of intermediate lobe peptides are discussed. PMID- 6287539 TI - Interaction of beta-endorphin, naloxone and dopamine: effects on melanocyte stimulating hormone secretion of amphibian pituitaries in vitro. AB - Neurointermediate lobes from amphibians (Rana pipiens) were incubated in Medium 199 containing dopamine, beta-endorphin or dopamine plus beta-endorphin. Dopamine inhibited melanocyte-stimulating hormone (MSH) secretion as measured by bioassay in hypophysectomized frogs, an effect which was transiently reversed by beta endorphin. The effects of endorphin were in turn partially suppressed by the opiate antagonist, naloxone hydrochloride. Cells treated with all three agents exhibited expanded rough endoplasmic reticulum and decreased secretory granule content, indicative of peptide release and new synthesis. Beta-Endorphin alone did not stimulate MSH secretion above control levels, and at one time period was seen to reduce MSH secretion. The findings indicate a complex interaction between beta-endorphin and dopamine directly upon MSH secretion at the level of the neurointermediate lobe. PMID- 6287540 TI - Neurotensin receptors in the rat bronchi. AB - The isolated, circular preparations of the left and right bronchi of the rat were examined for mechanical responses to neurotensin (NT) and other vasoactive peptides. NT caused concentration-dependent increases in the isometric tension of the unstimulated preparations, with an apparent affinity higher than for the cholinergic agonists but with considerably lower intrinsic activity. Pronounced tachyphylaxis to NT was observed. NT potentiated the atropine-sensitive increase in tension resulting from electrical field stimulation. Neither atropine nor methysergide abolished the response to NT in the unstimulated preparations. Bradykinin and to a lesser extent angiotensin II contracted the unstimulated preparations and both systemic peptides enhanced the cholinergic output in response to field stimulation. Substance P and VIP on the other hand were without effects in the stimulated and unstimulated bronchi. The results are consistent with the presence of receptors for NT on the presynaptic cholinergic terminals as well as on the post-synaptic smooth muscles of the rat bronchi. PMID- 6287541 TI - [Hypophyseal intratumoral apoplexy. Apropos of 5 cases]. PMID- 6287542 TI - [Endocrinological alterations in head injuries: alterations in cortisol and A.C.T.H. secretion]. PMID- 6287543 TI - [Spontaneous rupture of the stomach]. PMID- 6287544 TI - Lung cancer screening programs in Canadian uranium mines. PMID- 6287545 TI - Results of Memorial Sloan-Kettering lung project. PMID- 6287546 TI - Results of Mayo lung project: an interim report. AB - Screening for lung cancer by chest roentgenography and sputum cytology can be conducted by mail. Detection, localization, and treatment of early presymptomatic lesions are possible. The initial prevalence of lung cancer in men older than age 45 years who were heavy smokers was almost 1%. Rescreening at 4-monthly intervals detected 4.7 new cases of lung cancer per 1,000 subjects per year. Three times as many postsurgical stage I lesions were detected in the screened group as in the control group. Although survival seems to have been improved by screening, mortality from lung cancer is not yet significantly different in the 4-monthly surveillance and control groups. Longer follow-up is necessary for determination of the ultimate usefulness of screening. PMID- 6287547 TI - The contribution of uranium miners to lung cancer histogenesis. PMID- 6287549 TI - [Sudden deafness following a scratch-induced benign lymphoreticulosis (author's transl)]. PMID- 6287548 TI - Angiotensin-converting enzyme in serum and in bronchoalveolar lavage in sarcoidosis. AB - Angiotensin-converting enzyme (ACE) determinations were made in serum and in bronchoalveolar lavage fluid in 20 controls and in 28 patients with sarcoidosis. Serum ACE was significantly higher in patients with active sarcoidosis (54.3 +/- 19.0 SD nmol/ml/ min; n = 24) compared to controls (25.7 +/- 8.2; n = 20) or to patients with inactive sarcoidosis (23.6 +/- 7.3; n = 4). In contrast, ACE in bronchoalveolar lavage fluid was similar in nonsmoking controls (16.4 +/- 7.3 nmol/ml/min/macrophage; n = 8), smoking controls (10.4 +/- 11.9; n = 7); nonsmoking active sarcoidosis patients (16.7 +/- 14.6; n = 10), smoking sarcoidosis patients (17.9 +/- 8.4; n = 6) and inactive sarcoidosis patients (14.5 +/- 8.2; n = 3). Since ACE has been demonstrated by immunofluorescence in mononuclear phagocytes in granulomas, the authors speculate that macrophages recovered by alveolar lavage are not activated and do not reflect sarcoid alveolitis at the tissue level. PMID- 6287550 TI - [Iodide-perchlorate discharge test in children living in an area of endemic goiter (author's transl)]. PMID- 6287551 TI - [Islet cell tumors of the pancreas]. PMID- 6287552 TI - [Motor neglect of thalamic origin: report on two cases (author's transl)]. AB - Two cases of thalamic lesions with motor neglect are presented. The syndrome of motor neglect was complete in those cases with a) underutilization of left limbs, but good utilization upon verbal orders, b) loss of placement reaction, c) weakness of movement when hand was approaching the target, d) weakness of motor reaction to nociceptive stimuli. Those cases confirm that motor neglect exists after thalamic lesions and bring pathologic clues for topographic discussion. Motor neglect seems to be a particular case of partial unilateral neglect throwing some doubt on the hypothesis of a global trouble of hemispheric activation. Prevalence of left motor neglects suggests some linkage between propositional motility and language. One may suppose that in the right hemisphere language is able to have a vicarious action when spontaneous activation is lost; at the opposite, in the left hemisphere language and motility would be too linked to let this dissociation be generally possible. PMID- 6287554 TI - [Peripheral neuropathy due to N-hexane in a drug addict (author's transl)]. AB - After voluntary inhalation of a domestic solvent containing N-Hexane and N Heptane for three months, a 23-year-old woman developed motor deficit of the lower limbs, sensory symptoms and areflexia. Clinical disorders continued to progress after discontinuation of the intoxication, with a parallel aggravation of the E.M.G. disturbances. A nerve biopsy with ultrastructural study showed axon dilatation with accumulation of neurofilaments. The clinical, electrophysiological and pathological features of neuropathies induced by hexacarbon solvents are reviewed and their pathogenesis is discussed. PMID- 6287553 TI - [Flutter-opsoclonus: report on three cases (author's transl)]. AB - Three cases of spontaneous saccadic ocular movements are reported, each with one or several electrooculographic recordings. Case 1 is a typical ocular flutter during a myoclonic encephalitis with cerebellar signs. Case 2 is an ocular flutter occurring in the course of an acute inflammatory polyneuropathy with cerebellar signs after cytomegalovirus infection. Case 3 began with permanent dissociated opsoclonus, then conjugated opsoclonus and ended with vertical flutter in a patient suffering from bronchial carcinoma. While some definitions are unclear, clinical, electrooculographic and etiological data support a unicist point of view on flutter-opsoclonus. PMID- 6287555 TI - [Combination of a myopathy and involvement of the central and peripheral nervous system (author's transl)]. PMID- 6287556 TI - [Polymorphism of DNA in human populations using the method of hybridization after transfer]. PMID- 6287558 TI - The production in culture of metalloproteinases and an inhibitor by joint tissues from normal rabbits, and from rabbits with a model arthritis. II. Articular cartilage. AB - During the development of proliferative arthritis in the knee joints of rabbits, there was a large increase in the ability of articular cartilage explants to produce latent collagenase in culture. In parallel, the normally high levels of collagenase inhibitor produced by cartilage in culture fell, but active collagenase was never detectable. Characterisation of the collagenase and other proteinase activities produced by rabbit articular cartilage in culture showed that two activities could be separated by gel filtration, one with activities on gelatin and cartilage proteoglycan and the other degrading collage. Under the conditions employed in this paper no resolution of the gelatin and proteoglycan activities could be achieved. All the activities were in a latent form, activated by 4-aminophenylmercuric acetate (APMA), and inhibited by 1,10-phenanthroline or EDTA, but not by di-isopropylfluorophosphate (DFP), indicating that they are metalloproteinases. Characterization of the collagenase inhibitor showed a single peak of activity of apparent molecular weight of 28,000 on gel filtration. The inhibitor was sensitive to APMA and also inhibited other rabbit metalloproteinases, analogous to the system described for rabbit bone. The physiological significance of the synthesis by articular cartilage of proteinases that destroy connective tissue macromolecules and the presence of an enzyme inhibitor control system is discussed. PMID- 6287557 TI - The production in culture of metalloproteinases and an inhibitor by joint tissues from normal rabbits, and from rabbits with a model arthritis. I. Synovium. AB - During 5 days of culture, explants of normal rabbit synovium produced no active collagenase, negligible latent collagenase, but significant levels of free collagenase inhibitor. Synovium from joints exhibiting a proliferative arthritis produced greatly elevated levels of collagenase; the appearance of active enzyme in the medium during the second day of culture was associated with the disappearance of free inhibitor. Enzyme levels in the media correlated well with the arthritic status of joints, when explants were prepared up to 10 weeks after the induction of the model arthritis. Synovium from the contralateral joints of rabbits with unilaterally induced arthritis produced no active collagenase, but approximately one-third as much latent collagenase as found with arthritic joints. Enzymatic activities against gelatin and cartilage proteoglycan substrates were demonstrated in synovial culture media in addition to collagenolytic activity. Gel filtration showed that these activities were not due to a single enzyme, and further characterisation confirmed that the enzymes were metalloproteinases. The results are considered in the light of published data, and the involvement of metalloproteinases and their specific inhibitor in the development of arthritic lesions is discussed. PMID- 6287559 TI - A radial diffusion assay for plasma and serum deoxyribonuclease I. AB - A simple radial diffusion technique is described for assaying deoxyribonuclease I (DNase I) activity in whole human plasma and serum. Enzymatic activity was calculated from a standard curve constructed from the studies on the hydrolysis of calf thymus DNA by bovine pancreatic DNase of known concentrations. The assay was shown to be specific for nucleases which are active at neutral pH and require magnesium ions for activation. There was no significant difference in DNase I activity in plasma compared with that in serum from normal individuals (P greater than 0.05). The DNase I activity of 35 normal human sera was 26.1 +/- 9.2 ng/ml and ranged from 10.8-48.5 ng/ml. This technique may prove useful for the evaluation of DNase I activity in crude biological fluids and is not affected by the presence of enzymatic inhibitors and/or activators. PMID- 6287560 TI - Serum deoxyribonuclease I and clinical activity in systemic lupus erythematosus. AB - Serum deoxyribonuclease I (DNase I) activity in systemic lupus erythematosus (SLE) patients was shown to be lower than that of healthy laboratory personnel, rheumatoid arthritis, and scleroderma patients (P less than 0.001). The decrease in DNase I activity in SLE sera was not due to the effect of various autoantibodies or to heat labile DNase I inhibitor. A relationship between serum DNase I activity and active SLE was demonstrated. Patients with active lupus nephritis had the lowest levels of enzymatic activity. PMID- 6287561 TI - The effects of dexamethasone in vitro on the production of collagenase and inhibitor by synovial and cartilage explants from the joints of rabbits with a proliferative arthritis. AB - Using a rabbit model arthritis we have investigated the ability of dexamethasone to alter the production of collagenase and the specific metallo-proteinase inhibitor TIMP by explants of synovium and cartilage in vitro. The patterns of collagenase and TIMP production by untreated explants from arthritic joint tissues in culture were similar to those described previously [1, 2]. Dexamethasone dramatically altered the patterns of production of collagenase and TIMP. At a dose of 10 nM, or above, the patterns of production by treated synovium resembled those of normal rabbit synovium: collagenase production was suppressed and TIMP increased compared with untreated arthritic synovium. The levels of latent collagenase in cartilage also fell with increasing doses of dexamethasone and TIMP levels were higher, although normal levels were not reached. These experiments have been conducted as a prelude to testing the effects of various anti-rheumatic drugs in vivo, and attempting to correlate changes in clinical parameters with the subsequent production of collagenase and TIMP in vitro. The data are discussed in relation to the therapeutic use of corticosteroids and to their mode of action on joint tissues. PMID- 6287563 TI - [Lesion of the brachial plexus as a result of radiation therapy]. PMID- 6287562 TI - Lysosomal elastase: effect on mechanical and biochemical properties of normal cartilage, inhibition by polysulfonated glycosaminoglycan, and binding to chondrocytes. AB - Rheumatic joint destruction usually starts with the destabilisation of cartilage. Lysosomal elastase is a candidate effector of this process, since this enzyme is found at the site of cartilage erosion by rheumatoid synovial tissue. In order to prove this hypothesis we assessed the mechanical stability of cartilage during treatment by this enzyme in vitro. An indentation apparatus was used for this purpose and biochemical as well as microscopic techniques were used to supplement the results thus obtained. Our findings show that elastase irreversibly impairs the stability of cartilage by lysis of matrix proteoglycans without the help of additive enzymes. Collagen fragmentation played no significant role during elastase-induced destabilisation, while specific collagenase attacked the collagen network within the matrix only subsequent to the removal of proteoglycans. These findings suggest that elastase is a leading enzyme during proteolytic cartilage degradation. In addition polysulfonated glycosaminoglycan was found to reduce the mechanical effect of elastase on normal cartilage. It is therefore concluded that local inhibition of elastase promises therapeutic benefit during rheumatic cartilage degradation. Upon treatment of cartilage with elastase we observed this enzyme not only within the matrix under destruction but also bound to chondrocytes. These findings support the hypothesis that elastase plays a role on the matrix not only by direct degradation, but also by an indirect effect mediated through living chondrocytes. PMID- 6287564 TI - [Hypertrophic neuropathies beginning in infancy: a study of 3 cases (author's transl)]. AB - The genetical forms of hypertrophic neuropathies, inherited either as recessive or autosomal dominant trait, are classified, according to Dyck (1975), as HMSN type I, III, and IV. Sporadic cases are also reported. We studied three patients, one with autosomal recessive inheritance, and two without family history, who had the following common features: --onset of symptoms before the age to two years; - slowly progressive course; --peroneal muscular atrophy with absent tendon reflexes; --reduction of MCV and SCV; --decreased number of myelinated fibers; - schwannian cell hyperplasia, with onion bulb complexes formation; --absence of aspects of hypomyelination; --increased number of collagen pockets and denervated Schwann-Remak cells or processes. On light microscopy, multilamellated onion bulbs of large size were found in a very high percentage in case 1, while there were either simple in type or in a lower percentage in case 2. In the third, case, onion bulbs were recognized only on electron microscopy. It is known that in the various kinships affected with type I of HMSN, the pathological changes of peripheral nerves differ greatly. Therefore, despite early onset of symptoms and varying degree of severity of nerve changes, all three cases have been classified within the group of HMSN type I. The different severity of nerve damage may suggest the possibility of a genetical heterogeneity in this disorder. PMID- 6287565 TI - [Immune complexes and peripheral neuropathy: about two cases (author's transl)]. AB - Two female patients aged 64 and 54 were studied, affected by a progressive, distal sensori-motor mononeuritis multiplex. A nerve biopsy specimen from the first patient, suffering from chronic active hepatitis without detectable serum hepatitis B surface antigen (HBsAg) or antibody (anti-HBs), showed segmental demyelination and widespread inflammatory and necrotizing vasculitis and perivasculitis. The second patient, affected by a localized, caeseating tuberculous infection of the right kidney, refused to undergo a nerve biopsy. Serum levels of circulating immune complexes, detected by C1q BA binding method were strongly positive in both subjects; activation of the complement system (decreased serum level of C3 and/or C4) was also present. The course of the peripheral neuropathy was progressive and worsened in the first patient whose circulating immune complexes were always detectable. On the contrary the second patient showed improvement after the nephrectomy, which coincided with a quick disappearance of the circulating immune complexes. The findings may suggest in these two patients an immune complex mediated pathogenetic mechanism like the one recently accepted for the peripheral neuropathies in the course of chronic hepatitis B. PMID- 6287566 TI - [Paget's disease]. PMID- 6287567 TI - [Bone metastases of cystic adenoid carcinoma of the parotid. Apropos of a personal case and review of the literature]. PMID- 6287568 TI - Forms of binaural summation and the implications of individual variability for binaural hearing aids. AB - The advantages which a two-ear system has over a one-ear system appear to be manifold, but not all understood in detail. All probably play some role in the generally recognised advantage of binaural aids in offsetting auditory disabilities. Several possible forms of binaural summation are distinguished here and their possible role discussed. Experimental results with normally-hearing listeners and with aid users show that binaural summation is likely to lead to gain settings of about 6 dB lower than with otherwise equivalent monaural amplification. In some cases this may be a direct cause of benefit. However individuals differ systematically in the extent to which they show this effect, which may be relevant to prognosis for binaural aiding. Evidence is reported of an association between binaural summation and binaural advantage. Individuals also differ systematically in binaural summation for uncomfortable loudness level. PMID- 6287569 TI - [Near-drowning in an adult: favorable course after a 20-minute submersion]. AB - Cardiopulmonary resuscitation was successful in a healthy 29-year-old woman who had been submerged for 20 minutes in water at 10 degrees C. The evolution was characterized by the development of a multifactorial ARDS (secondary drowning) and sepsis caused by Aeromonas hydrophila and Acinetobacter anitratum. Fibrosing alveolitis caused a restrictive syndrome with severe mechanical impairment and transitory therapy-resistant hypoxemia. It is suggested that prolonged submersion in cold water is also a treatable and completely reversible condition in adults. In our patient without neurological sequelae the pulmonary function studies after 3 months show complete recovery from the mechanical impairment. After a follow-up period of 11 months only mild abnormalities of gas exchange persist. PMID- 6287570 TI - The myasthenic (Eaton-Lambert) syndrome associated with carcinoma. Enzyme induction as a possible mechanism of paraneoplastic syndromes. AB - A myasthenic syndrome associated with small-cell tumours of the bronchus and with autoimmune diseases (Eaton-Lambert syndrome) has been attributed to diminished probability that a nerve action potential will release acetylcholine (ACh) from terminals of cholinergic nerves (somatic motor and autonomic). This model derives from evidence for reduced quantal content of the transmitter released by a nerve impulse. The test procedure implies certain constancies of postsynaptic response. Abnormal responses to ACh-agonists indicate that receptor response is not normal. It is suggested that all previously described neuromuscular responses are compatible with new model: the subsynaptic apparatus produces excess acetylcholinesterase (AChE) which limits the endplate conductance changes produced by normal output of ACh. This model is supported by earlier ultramicroscopic studies which cannot be accounted for by the contemporary model. It is proposed that enzyme induction by peptide or immunoglobulin may also be responsible for other paraneoplastic syndromes. PMID- 6287571 TI - The distribution of oxidized nitrogen in urban air. AB - This paper describes the distribution of oxidized nitrogen compounds in urban air. The studies reported here have focused on NO, NO2, PAN and NO-3. These species have been monitored for periods of several weeks at a nmber of locations throughout the U.S.A. Oxidized nitrogen distribution is discussed in terms of the daily patterns of nitrogen pollutant concentrations, the fractional conversion of NOx to products and the diurnal profile of the conversion fraction, and lastly, the contribution of the individual compounds to the overall oxidized nitrogen burden. Site to site variations in the distribution are explained by photochemical conversion, distance between the site and the source region, and the proximity of local NOx emission sources. PMID- 6287572 TI - Nucleotide sequence of the p21 transforming protein of Harvey murine sarcoma virus. AB - Harvey murine sarcoma virus is a retrovirus which transforms cells by means of a single virally encoded protein called p21 has. We have determined the nucleotide sequence of 1.0 kilobase in the 5' half of the viral genome which encompasses the has coding sequences and its associated regulatory signals. The nucleotide sequence has identified the amino acid sequence of two additional overlapping polypeptides which share their reading frames and the carboxyl termini with p21 but which contain additional NH2-terminal amino acids. PMID- 6287573 TI - Nucleotide sequence of the oncogene encoding the p21 transforming protein of Kirsten murine sarcoma virus. AB - The transforming protein of Kirsten murine sarcoma virus (Ki-MuSV) is a virally encoded 21-kilodalton protein called p21 kis. The sequences encoding p21 kis were genetically localized to a 1.3-kilobase segment near the 5' end of the viral genome by assaying the capacity of a series of defined deletion mutants of molecularly cloned Ki-MuSV DNA to induce focal transformation of mouse cells. Nucleotide sequencing of a portion of this region has led to the identification of an open reading frame of 567 nucleotides coding for p21 kis protein. PMID- 6287574 TI - Phagocyte impotence caused by an invasive bacterial adenylate cyclase. AB - For unknown reasons, humans infected with the bacterium Bordetella pertussis are exceptionally vulnerable to secondary infections. Bordetella species elaborate a soluble, heat-stable, and highly active adenylate cyclase. This enzyme is internalized by phagocytic cells and catalyzes the unregulated formation of adenosine 3',5'-monophosphate (cyclic AMP), thereby disrupting normal cellular function. This unusual phenomenon may explain Bordetella-induced aphylaxis and may prove to be useful for investigating a variety of cyclic AMP-governed processes. PMID- 6287576 TI - Regulation of circadian rhythmicity. AB - Daily rhythms in many behavioral, physiological, and biochemical functions are generated by endogenous oscillators that function as internal 24-hour clocks. Under natural conditions, these oscillators are synchronized to the daily environmental cycle of light and darkness. Recent advances in locating circadian pacemakers in the brain and in establishing model systems promise to shed light on the cellular and biochemical mechanisms involved in the generation and regulation of circadian rhythms. PMID- 6287575 TI - Human beta-globin gene sequences injected into mouse eggs, retained in adults, and transmitted to progeny. AB - Foreign gene sequences were retained in two adult mice (out of 62 analyzed) from fertilized eggs injected with a recombinant plasmid containing the human beta globin genomic region and the herpes simplex viral thymidine kinase gene. The intact human and viral genes were found in DNA of one of the animals and, in the other, at least part of the human globin gene was present. The latter individual transmitted these sequences to its progeny in a Mendelian ration. Thus, human DNA may be incorporated into the germ line of mice for in vivo studies of regulation of gene expression in development, genetic diseases, and malignancy. PMID- 6287577 TI - Induction of melanized cells from a goldfish erythrophoroma: isolation of pigment translocation variants. AB - Melanization was induced in some cells of a goldfish tumor cell line (GEM-81) by cultivating the cells in autologous serum. The melanized cells continued to proliferate in vitro and several clones were isolated that differed with respect to cell morphology and intracellular distribution of pigment. Some of the clones consisted of cells able to translocate their melanosomes in response to epinephrine, melatonin, or adenosine 3', 5'-monophosphate. PMID- 6287578 TI - Stereoselective antagonism of phencyclidine's discriminative properties by adenosine receptor agonists. AB - Rats trained to discriminate between phencyclidine and saline vehicle were used to test various agents for their ability to mimic or block the phencyclidine cue. ketamine, dexoxadrol, tiletamine, and phencyclidine analogs were found to mimic phencyclidine's behavioral effects. Treatment with the adenosime receptor agonists N6-cyclohexyladenosine and L-phenylisopropyladenosine blocked the discriminative properties of phencyclidine. These results suggest that adenosine receptor agonists might be useful in treating phencyclidine-induced psychosis. PMID- 6287579 TI - Precipitated withdrawal by a benzodiazepine receptor antagonist (Ro 15-1788) after 7 days of diazepam. AB - Baboons implanted with intragastric catheters were given diazepam (10 milligrams per kilogram of body weight) twice daily for 45 consecutive days. On days 7 and 35, they were given intramuscular injections of the benzodiazepine receptor antagonist Ro 15-1788. Mild and intermediate withdrawal signs, including retching and vomiting, were observed after 7 days of diazepam, and more frequent and intense withdrawal signs, including tremor and convulsion, occurred after 35 days of diazepam. With the termination of the diazepam injections after 45 days, a mild to intermediate withdrawal syndrome was observed over the next 15-day period. PMID- 6287580 TI - Numbers of receptor sites from Scatchard graphs: facts and fantasies. AB - Data for ligand and receptor binding presented in the format of a Scatchard graph are compared with the same data shown as bound ligand plotted against the logarithm of free ligand. From this comparison it is apparent that extrapolations in the Scatchard graph to yield total number of receptor sites are generally not correct. PMID- 6287581 TI - Naloxone antagonism of the thermoregulatory effects of phencyclidine. AB - Phencyclidine elicits hyperthermia at low doses and hypothermia at high doses in rats. Naloxone antagonizes both effects. Phencyclidine's effects on thermo regulation are probably mediated by an interaction with a mu opiate receptor. PMID- 6287582 TI - [Lipoproteinemia throughout puberty]. AB - In prepubertal children, total cholesterol and HDL cholesterol concentrations are identical in both sexes, while total cholesterol/HDL cholesterol and LDL + VLDL cholesterol/ HDL cholesterol ratios are slightly higher in girls. During puberty, total cholesterol concentrations decrease slightly more in boys than in girls; HDL cholesterol concentrations remain unchanged in girls but decrease significantly in boys. When puberty is completed, in comparison to prepubertal values, total cholesterol/HDL cholesterol and total cholesterol/LDL + VLDL cholesterol ratios are significantly increased in boys, while they are unchanged in girls. During puberty, rises in triglyceride concentrations are significant in boys only. PMID- 6287583 TI - [Physical activity in the management of hyperlipoproteinemia]. AB - Increasing patients' physical activity significantly enhanced the efficiency of nutritional management in three forms of hyperlipoproteinemia: hypercholesterolemia, hypertriglyceridemia, and hypercholesterolemia with hypertriglyceridemia. Although total cholesterol concentrations are only moderately decreased by physical exertion, HDL cholesterol concentrations are increased, resulting in a significant fall in total cholesterol/HDL cholesterol ratios. In addition, triglyceride concentrations decrease significantly. PMID- 6287584 TI - [Serum angiotensin converting enzyme activity in sarcoidosis. A review of seventy cases]. AB - The serum angiotensin converting enzyme (ACE) activity was assayed in seventy untreated patients with sarcoidosis. Results were considered abnormal if they exceeded 34,5 U/ml. (SEM + 250). Increased ACE activity was found in 55,7% of the patients. Most of these patients had either Stage II or disseminated disease. Significant decreases in ACE activity always occurred during remission, whether spontaneous or induced by corticosteroids. In most patients with persistant roentgenologic features ACE activity was still high six months after onset. No correlations were found between serum ACE activities and lymphocytosis in bronchial washing samples. We conclude that, while serum ACE activity assays are not of significant diagnostic utility in patients with sarcoidosis, they are helpful for monitoring the course of the disease. PMID- 6287585 TI - [Blood group ABO (H) cell surface antigens in bladder tumours invading the chorion. Prognostic value of their determination by immunofluorescence. Retrospective study of one hundred and twenty-five cases]. AB - The authors have studied 125 stage A bladder tumours with a follow-up period of at least 5 years. The ABH cell surface antigens were collected by a double-layer immunofluorescence technique which has already been described. All the cross sections were read by the same pathologist who knew nothing about the clinical developments. The results were divided into 3 groups : (I) all the tumour cells had retained their surface antigens. (II) Only some cells had retained surface antigens. (III) All cells were negative. Study of the development of the 3 groups shows that : in group I, all the patients (11) are alive 5 years later, and only one has presented with a lesion evolving towards infiltration. In group II (32 patients), only 38 p. cent of the patients are still living 5 years later, and 65 p. cent developed towards muscular infiltration. In group III (77 patients), 27 p. cent are alive after 5 years and 80 p. cent have developed towards muscular infiltration. Homogeneous retention of the surface antigens would therefore seem to be a favourable factor in the prognosis, even in the case of a tumour which has already infiltrated the chorion. The significance of the loss of surface antigens is less clear-cut, but is on the whole adverse. The significance would, however, probably be enhanced by study of the mucosa distal from the lesion, and by marking the substances which determine the blood groups. PMID- 6287586 TI - [Comparative study of the bacteriologic value of samples taken by transtracheal aspiration or fiberoptic bronchoscopy]. AB - To evaluate two methods of obtaining lower respiratory tract secretions, fiberoptic bronchoscopy (FB) and transtracheal aspiration (TA) were performed in ten patients. FB used an open-end brush-in catheter. TA was performed as described by Pecora. TA gave significantly better sensitivity and specificity (3/5 and O false positive results respectively) than FB (2/5 and 5 false positive results). TA gives specimens which reflect the bacterial flora of the lung more reliably than those taken during FB. PMID- 6287587 TI - [Relationship with height- and weight-characteristics]. AB - Blood pressure (BP) has been measured by one observer among 837 male healthy subjects, aged 18. As it has been shown in previous studies, there is a strong correlation between BP and height, weight, overweight indices, and arm circumference. The correlations of height and arm-circumference with BP are not fully explained by the weight. It seems to be, at this age, an association of BP with body dimensions, and not only with fatness. The strong association of BP with the lean body mass confirms this fact. It seems finally that BP level at the beginning of adulthood is, in a large extent, determined by body dimensions and the development of the osteomuscular system, and much lesser by adiposity. On the other hand, evolution during adulthood depends much more upon variation of the weight, i.e of the fatness. PMID- 6287588 TI - [Frequency of routine examinations in detection of carcinoma of the uterine cervix and carcinoma of the breast]. AB - The prevention of carcinoma of the uterine cervix depends on routine examinations. They should start soon after the first experiences of sexual intercourse then be carried out once every two years in the form of a smear until the age of thirty, then every three years between 40 and 55 years. After 55 years, one should advise a cervical smear every 5 years, although the probability of detecting a carcinoma of the cervix is very small if the woman has until then been well followed up. However, when there is a high risk, in particular during oral contraception, smears should be taken once a year. This type of detection is possible, there are enough doctors, and each should carry out 23 smears a month, similarly there are enough cytologists. Finally, the economy made by not having to treat lesions detected early by this method easily covers the cost. The detection of carcinoma of the breast requires auto-palpation of the breast and mammography. A baseline mammography should be performed for reference purposes between 35 and 40 years. In the absence of a risk factor, and if the mammography shows low density breasts (P1 or N1 according to Wolfe's classification), one mammography every five years until the age of 50 years is suggested. In high risk cases, or if the breasts are denser (N2 or Dy), an annual ray is recommended using sensitive films which permit reduction of the dose received by the breast. After 50 years, annual mammography is strictly necessary until the age of 70 years. Applying these data to the French population, one may consider that four million women will be concerned each year by mammography (15% of the female population). Considering the reduction in costs by treatment of carcinoma of the breast at an early stage, each examination would cost less than 45 F. PMID- 6287589 TI - [Unexpected generalized amyloidosis discovered on biopsies. Value of Wright's method for etiologic diagnosis. Three case-reports]. AB - Three cases are reported in which unexpected generalized amyloidosis was disclosed by biopsies. The authors emphasize the value of precise analysis of the deposits and particularly of Wright's method for studying their staining properties. This method can guide investigations towards one of the two main etiologies of amyloidosis (immunoglobulinic or AA). PMID- 6287590 TI - [Fulminant infection in a splenectomized patient with Hodgkin disease. Fatal pneumococcal meningitis in a cured patient seven years after splenectomy]. AB - Ten years ago, because of the increasing number of patients recovering from Hodgkin disease, some authors advocated systematic laparotomy with splenectomy for more accurate staging in order to adjust therapy with more precision. This attitude now seems questionable because of the occurence of several complications, particularly of life-threatening infections, which can be fatal even after recovery from Hodgkin disease. The case of a twenty-three-year-old male patient with Hodgkin disease stage III A, histological grade I, is reported. MOPP chemotherapy, splenectomy and cobalt radiotherapy were followed by complete recovery. Seven years later the patient died from fulminant pneumococcal meningitis. A review of previously published cases show that prolonged prophylaxis with oral penicillin is mandatory in splenectomized patients. Immunization with the pneumococcal polysaccharide vaccine should be attempted even though the long-term results are still unknown. PMID- 6287591 TI - [Malignant transformation of fibrous dysplasia. About a case review of the literature]. AB - A case of chondrosarcoma arrising from polyostotic form of fibrous dysplasia coming after 35 years is reported. About this new occurence, a review of the 61 cases of the literature is effectuated. Sarcomatous transformation is a rare complication of fibrous dysplasia. Its frequency change with the authors, because of lack of proof of the preexistence of fibrous dysplasia. She occur specially on polyostotic fibrous dysplasia. This complication is severe, death following before two years. In the literature several etiological factors are discussed: repeated X-rays, radioscopic examinations; cartilaginous nodules. In our case we means that the sarcoma arrise on a chondroblastic bone healing. PMID- 6287592 TI - [Pain and inflammation of the foot in metabolic osteopathies : new causes of algodystrophy (author's transl)]. AB - Three observations of patients with algodystrophia of the foot are reported. Biologic disorders suggestive of osteomalacia were found in two patients who recovered after receiving vitamin D. Cushing disease was diagnosed in the remaining patient ; again algodystrophia disappeared after specific therapy was given. The few similar cases published in the medical literature are reviewed. Attention is drawn to their misleading nature. Most published cases occurred with osteomalacia or endocrinic osteopathy. Such conditions should be looked for in all patients with apparently primary algodystrophia of a lower limb. PMID- 6287593 TI - [Acute primary ulcer of the small intestine. Present concepts (author's transl)]. AB - Acute primary ulcer of the small intestine is an exceptional occurrence. Diagnosis is established only after a complication occurs. In most instances, this complication is perforation of the gut. Acute primary ulcer of the small intestine is accurately defined by specific histologic criteria. It can be unequivocally distinguished from the many other causes of spontaneous perforation of the small intestine. Experimental studies, although numerous, have not improved understanding of pathophysiology. Nosologic classification remains unclear. Many features are similar to those encountered in acute peptic ulcer but stress is usually absent. Other features resemble those of transient necrotizing enterocolitis ; acute primary ulcer of the small intestine may be a very localized form of this latter condition. Ischemia seems to be the most significant factor. However, for an ulcer and finally perforation to occur, ischemia must probably be associated with a number of other factors. PMID- 6287594 TI - [Congenital mesoblastic nephroma. Study of five cases and review of the literature (author's transl)]. AB - Mesoblastic nephroma (individualized by Bolande in 1967) is a congenital renal tumor sometimes locally and regionally aggressive but never giving metastases, in contrast with nephroblastoma. It almost always occurs in the newborn. Therefore, every renal tumor discovered before twelve months must evoke mesoblastic nephroma. No preoperative adjuvant treatment must be used in infants under six months old. Considering five personal observations and review of literature, the authors expose the main etiologic and clinical characteristics of the disease, the gross and microscopic features which allow its diagnosis and the histogenetic theories of this curious lesion, probably border-line between neoplasias and dysplasias of renal parenchyma. PMID- 6287595 TI - [The fuzzy set theory as a biomedical diagnostic aid (author's transl)]. AB - In this paper, the theory of fuzzy sets and fuzzy logic is approached as a diagnostic aid for practitioners and biologists. A few theoretical considerations are followed by two practical applications. The first is a modification of the WHO-Fredrickson classification of hyperlipoproteinemias, with the introduction of secondary or minor types corresponding to everyday medical practice. The other is an attempt at classifying the four most prevalent anemias encountered in internal medicine. Recent studies have shown that the various parameters should be grouped on a hierarchical basis. PMID- 6287597 TI - [Evaluation of a new antiemetic, alizapride, in cancerology (author's transl)]. AB - The antiemetic effect of alizapride was evaluated in 50 patients with advanced cancer undergoing chemotherapy with cis-platinum. Our results suggest that alizapride, in a dose of 800 or 900 mg, significantly alleviates nausea and emesis. In a preliminary study, we had established that, without antiemetic therapy, the same patients all suffered from emesis. On the whole, alizapride was well tolerated ; no major side-effects were recorded. PMID- 6287596 TI - [Partial monosomy 20q : a new syndrome. Regional assignment of the ADA locus on 20q132 (author's transl)]. AB - A karyotype 46,XY,20q-(q13 leads to qter) was found in an infant with severe mental deficiency, epilepsy, and the following dysmorphic features : upward slanting palpebral fissures, hypoplastic nasal bridge, bulbous nose, long philtrum, microretrognathia, and aplasia of the middle phalanx of fingers and toes. Adenosine deaminase activity was within the hemizygous range, permitting regional assignment of the ADA locus to 20q13 leads to qter. PMID- 6287598 TI - [Long-term results of urinary rehabilitation in acquired infantile paraplegia (author's transl)]. AB - The urinary rehabilitation of children suffering from acquired paraplegia is here the subject of a retrospective study of 29 patients observed over a long period. The study shows that the detrusor-sphincter balance cannot be exploited on a personal level before the age of 10 ; bacteriuria and persistent residue after the initial period may ultimately disappear under discontinuous bacteriostatic treatment and thanks to the acquisition of autonomy in relation to the residue ; out of the total of 29 patients, the authors only noted 3 morphological and functional deteriorations requiring cutaneous diversion by Bricker's method. Today, the number would have been even smaller thanks to better knowledge of the striated sphincter syndrome, which accounted for 2 of the 3 cases noted. At last in the case of 16 of the 20 initial results considered as favourable, in other words in a little over 3 out of 4 patients, the social and uro-nephrological situation is holding steady, and even improving. PMID- 6287599 TI - [Diffuse oxalosis in a 62 year old man (author's transl)]. PMID- 6287600 TI - [Cryptic miliary tuberculosis. Two case-reports (author's transl)]. AB - The term cryptic miliary tuberculosis designates a particular form of hematogenous tuberculosis in which the usual diagnostic criteria, especially the roentgenographic ones, are lacking. Two cases are reported. Initial clinical manifestations were prolonged fever, poor general condition, low blood pressure, low serum sodium, mild hepatic biological disorders, and myocarditis. No other radiological or biological features, suggestive of tuberculosis were found. In the first patient, the various visceral tuberculous localizations occurred under specific treatment, although the latter was otherwise effective. In the second patient, they were identified by pathological examination. The incidence of these occult forms of tuberculosis, where diagnosis is often established too late, and sometimes only postmortem, is not negligible. This possibility must be borne in mind in severe infectious conditions without obvious etiology, particularly when the following features are present : inadequate secretion of ADH, which often indicates latent encephalitic and meningitic involvement where CSF anomalies may be incomplete or lacking ; leucopenia or pancytopenia ; biological hepatic anomalies ; exceptionally, such as in the two cases described, cardiac involvement. Among diagnostic procedures, liver biopsy is of the utmost importance. Mere diagnostic presumption is sufficient to warrant initiation of antituberculous therapy. This ensures survival and, paradoxically, allows delayed overt clinical manifestations to develop. PMID- 6287601 TI - [IgM myeloma (author's transl)]. PMID- 6287602 TI - [Recent advances in atopic dermatitis (authors's transl)]. PMID- 6287603 TI - [Etiologic aspects of Reiter syndrome. Role of yersinial infections (author's transl)]. AB - Reiter syndrome occurs as a reaction to various infections, either by intracellular microorganisms (Chlamydia, Mlycoplasma) or by intestinal bacteria (dysenteric bacilli, Salmonella, Campylobacter, Klebsiella, Yersinia). Four personal observations of Reiter syndrome following yersinial infections (Yersinia enterocolitis in two cases and Yersinia pseudotuberculosis in the two other cases) are reported. Data concerning the yersinial etiology of Reiter syndrome are reviewed. PMID- 6287604 TI - [Bone lesions in chronic lymphoid leukemia (author's transl)]. PMID- 6287605 TI - [Joint involvement in hyperlipoproteinemia (author's transl)]. PMID- 6287606 TI - [Urinary calcium and phosphate during fasting. Effect of various hormonal factors (author's transl)]. AB - Obese patients who fasted during seven days were divided into four groups according to whether they received triiodothyronine (100 to 150 microgram per day), calcitonin (1 MRC unit per day), both T3 and calcitonin, or neither of these two hormones. Urinary calcium and phosphate were increased in patients receiving T3 or calcitonin. T3 given with calcitonin did not increase the calcitonin-induced calciuria but did increase the phosphaturia. The authors suggest a possible explanation for these findings. No changes in serum parathormone were recorded during fasting acidosis. PMID- 6287607 TI - [A study of three potential antidepressants (amineptin, mianserin and nomifensin) on the cardiovascular system and isolated intestine (author's transl)]. AB - We studied the effects on the cardiovascular system and isolated intestine of three potential antidepressants : amineptin, nomifensin and mianserin. 1) No animals died in spite of doses well above the therapeutic range, evidencing the good tolerance of the compounds. 2) Effects on heart rate were variable; the only significant effect was cardioacceleration seen with mianserin in doses above 5 mg/kg. 3) The only significant effect on the auriculoventricular refractory period was recorded with mianserin. 4) Amineptin was the only compound that increased blood pressure. 5) Auriculoventricular conduction was slowed with mianserin in doses above 10 mg/kg. 6) On isolated rat duodenum, mianserin was spasmogenic in concentrations above 100 microgram/100 ml. 7) in comparison to tricyclic antidepressants, these three compounds carry only few risks of adverse side-effects. PMID- 6287608 TI - [Acute poisoning with niflumic acid. A retrospective study of cases seen at the Paris Poison Control Center, 1974-1981 (author's transl)]. PMID- 6287609 TI - [Parotid tumours. Only 31% of mixed tumours. In one hundred and seventy-five parotidectomies (author's transl)]. AB - This article reports on an analytical study into the etiological aspects of 175 parotidectomies carried out in the Lebanon. In comparing our results with the classical data, we reached a certain number of interesting conclusions. Our study confirms classical breakdown of parotid tumours with 80% benign and 20% malignant. We also confirm the rise in the incidence of malignancy in line with age, and its classical predominance in male patients. Classically, the benign tumours can be divided into 80% mixed tumours, 8% warthin, and 6 to 7% other rare tumours. Our data concerning this breakdown of benign tumours is very different. We found 31% of mixed tumours, which is in line with the figure given by A. Palva. We can therefore conclude that mixed tumours are not the most common form of benign parotid tumours and that the so-called rare tumours account for 69% benign tumours. They include hemangiomas, tubercles, salivary cysts, chronic parotidits and Warthin's tumours. Another difference between the conclusions of our study and classical data concern warthin tumours, which account fort 18% of cases as compared to only 6% in the classical data. We can therefore conclude that the so-called "rare benign tumours of the parotid" show a far higher incidence in our country than the classical 6%, and in fact come far closer to 50% of all cases of benign tumours. PMID- 6287610 TI - [Study of cellular immunity using Rebuck's skin-window (author's transl)]. AB - Value of a modification of Rebuck's skin-window in cellular immunity studies. The mononuclear/polynuclear ratio is determined at given time-intervals. The increase in the number of macrophages as well as their kinetics can be pictured on a diagram, comparatively to the corresponding results in controls. This test appears to provide a means of assessing the activity of a malignant disorder. PMID- 6287611 TI - [Relationship between bronchial and ileal carcinoma (author's transl)]. AB - This article concerns a patient who underwent surgery, first for carcinoma of the right upper pulmonary lobe, then, ten months later, for carcinoma of the ileum revealed by an occlusive episode. At the present time, the patient is still alive, and receiving complementary radiotherapy and chemotherapy. the authors emphasize the originality of their observation, as well as the significant relationship between these two forms of apparently primary bronchial and ileal tumors. PMID- 6287612 TI - [Colonic involvement in systemic mastocytosis (author's transl)]. AB - The most common gastrointestinal lesions encountered in systemic mastocytosis are the gastric, duodenal and small bowell lesions. The authors present a case of diffuse colonic involvement diagnosed on a double contrast examination. The radiological signs correlated with the endoscopical and histological findings are due to a submucosal edema and a polymorphic cellular infiltrate. The involvement of the colon is probably more frequent then previously thought. It could explain the diarrhea encountered in some patients affected by this disease. PMID- 6287613 TI - [Renal manifestations in Hodgkin disease. Two case-reports with a review of the medical literature (author's transl)]. AB - The authors report on two patients in whom Hodgkin disease was revealed by renal complications: nephrotic syndrome in one patient and renal failure with enlarged kidneys secondary to malignant infiltration in the other. In comparison to other lymphomas, renal infiltration is infrequent in Hodgkin disease. Clinical manifestations of renal infiltration are delayed. Renal failure with enlarged kidneys, which is very uncommon, carries a poor prognosis and is often associated with involvement of the lungs. Nephrotic syndrome in Hodgkin disease may be secondary to one of the three following causes: renal vein compression, renal amyloidosis, or a paraneoplastic syndrome. The latter is the most common cause and is probably ascribable to immunologic disturbances. Among the other complications, infection and the renal consequences of metabolic disorders are mentioned. PMID- 6287614 TI - [Results of attempted isolation of Chlamydia trachomatis in 2,000 specimens (author's transl)]. AB - Chlamydia trachomatis was isolated in cultured L cell lines. 29.5% of 1115 specimens from the male urethra were positive; specimens included 13 samples of semen, with 7 positive results. 39.9% of 771 specimens from the female genital tract were positive. In this group, 46 samples (with 17 positive results) were either biopsy specimens from the Fallopian tubes or their adhesions, or samples from culdocentesis. The remaining samples came from cervical swabs. The difference in the prevalence of positive results between male and female patients is statistically significant and may result from a better sampling technique in women. Isolation of chlamydia in pelvic specimens and in 19 out of 49 pharyngeal samples from neonates illustrates the main complications of chlamydial infection: sterility may result from salpingitis and neonatal pneumonia from contamination during birth. Among 57 specimens from the conjunctiva, 26 were positive. Isolation of chlamydia is the best diagnostic procedure in chlamydial infection: the serotypes of isolated strains should be determined. PMID- 6287616 TI - [Results of rhinoplasties. A computer study of 351 medical records]. AB - The authors studied the cosmetic results of 1380 rhinoplasties carried out over a period of 10 years by two surgeons, Professor A. Pech and Professor M. Cannoni. They used a classical reduction rhinoplasty technique. Different procedures were used for the tip of the nose depending on the individual indications, and the development of ideas over a period of time. Only 351 files, complete with pre- and post-operative photographs have been selected. The photographs have been analyzed on the basis of 70 criteria by an independent, young female ENT specialist, and the results of her analysis were fed to the computer. The study provides three types of answer. Firstly, the results were good in 69% of cases, and the greater the experience of the surgeon, the greater the success. Secondly, the different techniques for correcting the tip of the nose are analysed with good scores for Cinelli's technique (69%). The tip modeling procedures of Converse and of Lipsett have been abandoned owing to the high frequency of round tips. Finally, the different types of anomalies encountered are listed: 42% round tips, 20% asymmetrical tips, 17% residual humps, 11% of wide nasal bones, hooked noses and postoperative deviations. The authors then comment on the results, focusing on following points: the frequency of incomplete files (70%); the general improvement in the results with the gaining of experience, the decline in the proportion of hooked noses, also in line with experience, from 24% in the first 5 years to 7% after 10 years; the development of ideas concerning the techniques to be used in correcting the tip of the nose with an evergrowing place for those sacrificing the dome of the alar cartilages, such as the "Hockey stick" and a personal procedure derived from Pollot; wide nasal bones and failures in the tracing of the naso-frontal angle (64%) for which no solution is suggested; and, finally, postoperative nasal deviations. Since undertaking this study, the authors have successfully avoided this defect by using the external approach. PMID- 6287615 TI - ["Pericardial" friction rubs in pulmonary embolism (author's transl)]. AB - In a series of sixty-five patients with pulmonary embolism, the main clinical feature was a systolic and diastolic friction rub in three cases. Such friction rubs are found in 1 to 5% of patients with pulmonary embolism. The characteristics of the rub are identical to those of pericardial friction rubs. According to some authors, a friction rub indicates massive pulmonary embolism. However, a friction rub may be found in less severe cases if embolization involves the paracardiac lobes or segments. The rub may be caused by one or the other of the following mechanisms. Severe pulmonary embolism or a free blood clot floating in the right atrium or ventricle may produce a sound resembling a friction rub. More often, a genuine pleuro-pericardial or pericardial friction rub originates in an inflammatory reaction to a contiguous pulmonary infarct. It is important to establish accurate diagnosis of pulmonary embolism in spite of a pericardial friction rub because of the therapeutic implications. Anticoagulant therapy in sufficient dosage must not be withheld in this situation. PMID- 6287617 TI - [De-epiphysiodesis and old traumata of the growth apparatus (author's transl)]. AB - Basing themselves on eight cases treated over the past six years, the authors recount their experience of the operative elimination of partial closure of the epiphyseal plate. The closures were due to various causes and involved the lower extremities of the femur or tibia. Operative elimination alone was carried out in three cases, and associated with bone realignment in the remaining five. The results confirm the best prognosis for comparatively recent, localized peripheral epiphyseal plate closures. PMID- 6287618 TI - [Pulmonary infection with Pasteurella multocida in an immuno-deficient patient (author's transl)]. AB - A seventy-seven-year-old woman, under intermittent treatment with chlorambucil for chronic lymphocytic leukemia, developed acute pneumonia (diffuse interstitial pneumonitis) due to Pasteurella multocida. No direct traumatism had been caused by her cat. Bacteriologic study material was obtained by guided transtracheal distal bronchial brushing and washing. Therapy with tetracyclin was rapidly successful. Pasteurella multocida is an opportunistic organism which can be responsible for severe infections in high-risk patients. PMID- 6287619 TI - [Lymphomatoid granulomatosis with mesenteric localization. Nosologic problems (author's transl)]. AB - The authors report the case of a 70-year-old woman with a mesenteric tumor, apparently isolated. Histologic examination led to the diagnosis of lymphomatoid granulomatosis. A review of the literature found four previously published cases of lymphomatoid granulomatosis with mesenteric localization, and with or without pulmonary involvement. The general features of this affection are recalled. Special attention is given to forms without pulmonary involvement (Liebow granulomatosis) as well as to their relationship with polymorphic reticulosis. Separation of these two disorders, which depended mainly on a strict distribution of the various lesions, seems questionable. The authors tend to consider that both conditions belong to the same nosologic group, sincea pathological features are similar and borderline cases exist. PMID- 6287621 TI - [Endoscopic removal of a duodenal villous tumor. Review of the literature (author's transl)]. AB - A benign villous tumor of the second duodenum was treated by endoscopic excision. Fifty-four previously published cases are reviewed. Villous tumors of the duodenum are very uncommon. Presenting features include intestinal bleeding and/or occlusive symptoms and/or obstructive jaundice. Diagnosis is established by fiberoptic endoscopy and biopsies. Complete removal of the tumor is mandatory because of the high incidence of malignant changes (approximately 33%). Endoscopic excision is only possible in a small number of cases, when the tumor is small, benign, and pedunculated. Complete removal of the tumor ald unequivocal diagnosis of benign tumor on histological examination are required to withstand from further therapy. The best procedure is often surgical excision or segmentary duodenal resection. This is satisfactory for benign lesions or tumors with focal in situ malignant changes. Pancreatoduodenectomy should be considered for invasive malignant tumors. PMID- 6287620 TI - [Ulcerative and mutilating acropathy (author's transl)]. AB - The authors report the case of a 50-year-old patient with bilateral plantar chronic trophic ulcers. He was a chronic alcoholic and presented serious nutritional deficiencies. The etiological problems raised by sensitive neuropathies are reconsidered. After discussion, the authors relate their observation of ulcerative and mutilating acropathy to a sporadic Thevenard syndrome. PMID- 6287622 TI - [Newer diagnostic procedures for chlamydial diseases (author's transl)]. AB - Chlamydiales are bacteries showing a growth cycle unique among procaryotes. The two species Chlamydia trachomatis and Chlamydia psittaci are genetically very distant and their pathogenicity for man is very distinct. Human chlamydia infections by Chlamydia trachomatis are diseases chiefly sexually transmitted and their epidemiological importance is growing. The relationship between chlamydial infections, Reiter disease, and cat scratch disease are discussed. The various laboratory diagnostic procedures are reported, including the techniques and their indications; the method of choice is in the majority of cases the isolation of Chlamydia on cell culture. PMID- 6287623 TI - [A review of ten cases of aldosteronoma. Diagnostic and therapeutic conclusions (author's transl)]. AB - Ten patients with primary hyperaldosteronism resulting from a functioning adrenocortical adenoma were studied. The high incidence of hypertension and/or renal disease in the families of these patients suggests that hereditary factors play a part in the genesis of aldosteronomas. In patients with hypertension and hypokalemia the diagnosis of primary hyperaldosteronism is established by measurements of serum renin activity and serum aldosterone concentrations under normal conditions and during pharmacodynamic tests. The adenoma is demonstrated by radioactive iodocholesterol scintigraphy and computerized tomography. In nearly two-thirds of the patients, unilateral adrenalectomy is followed by complete definitive recovery. PMID- 6287624 TI - [Value and diagnostic significance of serum lactic dehydrogenase in internal medicine (author's transl)]. AB - Among 2175 patients seen over the last three years in a non-specialized department of internal medicine with no intensive care unit, 100 had supranormal serum lactic dehydrogenase activities. These patients' case-reports have been analyzed. Nearly half the patients (47/100) had a malignant disease (cancer or hemopathy). Among the remaining patients, 19 had a hepatic disorder (alcohol hepatitis in 10, viral hepatitis in 8, and isoniazide hepatitis in 1), 7 had a heart disease (heart failure with hepatomegaly in 5, myocardial infarction in 2), and 27 had various other conditions (including hemolysis in 6 and polymyositis en 3). The value of serum LDH assay is obvious in situations other than acute conditions such as myocardial infarction of pulmonary embolism; these are better known and have not been studied here as their prevalence was low among the patients enlisted in our study. In comparison to other enzymes (alkaline phosphatase (AP), gamma-glutamyl transpeptidase (GGT), transaminases (GOT, GPT) that were also routinely assayed in our patients, abnormal serum LDH activities are much less common and their significance is quite different. An increase in serum and their significance is quite different. An increase in serum LDH activity indicates a serious condition, often with a fatal outcome. The "various other conditions" group includes patients with hemolysis, hepatitis and myositis; the other patients in this group either had severe infectious diseases or died suddenly in the first few days of their hospitalization before diagnosis had been established. Each etiologic group has been analyzed to asses the characteristics of patients with increased LDH activity according to each etiology. Analysis of coincident abnormalities of the other enzymes listed above shows marked differences between etiologic groups; diagnostic accuracy can thus be enhanced in certain conditions. Most patients with malignancies had poorly differentiated tumors, with metastases: 28 had an epithelial tumor, with hepatic and/or bone metastases in 23 cases, 5 had cancer of the liver, 10 had a malignant hemopathy (2 lymphomas, 5 myeloproliferative syndromes, 3 acute leukemias), and 4 had a sarcoma. Cancer of the lung is the most common malignancy (10 cases) and may be responsible for increased serum LDH activity even in patients without metastases. Serum LDH assay is of value for monitoring the course in patients with initially increased activities as it falls under effective therapy and rises during exacerbations. PMID- 6287625 TI - [Difficulties in the pathological diagnosis of Garre's neurinoma (glandular schwannoma (author's transl)]. AB - Evidence from a case studied with electron microscopy suggests that tumors diagnosed up till now as Garre's neurinoma (glandular schwannoma), without ultrastructural proof, may not in fact originate in Schwann cells. None of the characteristics of Schwann cells could be demonstrated in the fusiform cells of the tumor. The medical literature is reviewed. PMID- 6287626 TI - [Breast carcinoma with multinucleated reactive stromal giant cells (author's transl)]. AB - Breast carcinoma with multinucleated reactive stromal giant cells is a rare histological type. The number of published cases, to which we add two new cases, is too small for it to be possible to reach a conclusion as to whether this type of complaint, for which the authors put forward morphological criteria, should or should not be considered as an anatomoclinical entity. The diagnosis can be made as soon as cytological examination has shown a large number of multinucleated giant cells associated with carcinomatous cells. Ultrastructural study confirms the macrophagic origin of the giant cells but does not show any macrophagic activity. PMID- 6287627 TI - [Bullous pemphigoid in children (author's transl)]. AB - A case of bullous pemphigoid (BP) in a five-year-old girl is reported. IgG deposits at the epidermal-dermal junction and high titres of serum IgG (1/10000) reacting with the epidermal-dermal junction of rabbit esophagus were the two criteria that led to the diagnosis of BP. Clinically and histologically this case had at first been diagnosed as dermatitis herpetiformis or benign chronic bullous disease of childhood. This emphasizes the importance of immunological criteria for classifying chronic bullous diseases in children. Our patient did not respond to sulfones and prednisone was given for thirteen months. No relapse occurred during the first year following prednisone withdrawal. Our observation is the twenty-first case of immunologically proven BP in childhood. The twenty-one cases are reviewed in order to analyze the features of the disease in children. PMID- 6287628 TI - [Results of great nerve trunk grafts]. AB - The authors describe the results of 60 cases of trunk repairs, carried out by the same surgeon, and with a follow-up period of 18 months or more. The grafts were autologous and in most cases the donor was the saphenous nerve. The grafts were placed under microscope. The recipient sites were the median nerve (19 cases), the ulnar nerve (16 cases), the radial nerve (10 cases), the sciatic nerve (8 cases), the musculo-cutaneous nerve (6 cases) and the spinal nerves (1 case). After a description of the technique used, the results are analyzed in accordance with the classical criteria of Highet and Holmes, to which has been added the notion of harmony of movement. The overall evaluation has been carried out in accordance with the following scale: two-point discrimination less than 15 mm + M4 or M5 + harmony = very good; two-point discrimination impossible to interpret, protective sensitivity S2 + M4 or M5 + harmony - good; crude protective sensitivity (S1) + M3 or less X average. Of the 60 cases, 21 ranks as very good and 22 as good, together giving an aggregate 72% of the total, with 12 cases ranking as mediocre and 5 failures. The incidence on these results of the following factors in then considered: the patient's age, the nerve damaged, the level of the injury in a given nerve, the time intervening between the initial lesion and the repair, the length of the nerve injury, the number of fascicles, the quality of the tissues and the surgeon's experience. It would seem that the best chances of success involve: a patient aged under 30, a short intervening period (less than 6 months between accident and repair), an injury less than 5 cms long, the maximum number of fascicles, good quality tissues and an experienced surgeon. PMID- 6287629 TI - [Dirofilariasis in humans. Report of a case with localization in the cheek (author's transl)]. AB - Cutaneous dirofilariasis usually affects animals (dogs and cats) but can occur in human-beings. Occasionally, this cosmopolitan disease is seen in France. Our case report is the sixteenth registered in France. The patient, a forty-two-year-old man, was infected in Corsica. He presented with a tumor of the right cheek from which a pregnant adult Dirofilaria (Nochtiella) repens was extracted. PMID- 6287630 TI - [Hepatitis and hyperthyroidism (author's transl)]. AB - Two cases of hyperthyroidism with hepatic manifestations are reported. In both the clinical picture was misleading. In one case, hyperthyroidism was responsible for liver involvement. Hepatic manifestations of hyperthyroidism are both polymorphic and non-specific. Diagnosis should be considered only after outruling other possibilities. In the other case, cholestatic hepatitis developed independantly from hyperthyroidism. The resulting clinical picture was confusing. All patients should be checked for non-thyrotoxic liver disease which may be of prognostic and therapeutic significance. PMID- 6287631 TI - [Pericarditis and chronic myelocytic leukemia (author's transl)]. PMID- 6287632 TI - [Postoperative insulinotherapy. Methods and indications (author's transl)]. AB - Postoperative insulinotherapy is widely used not only for diabetics, but also for patients suffering from glycoregulation anomalies induced by stress and parenteral nutrition rich in glucose. The authors point to recent progress in this field resulting from the development of new insulins and new procedures, such as continuous intravenous infusion with a constant flow. The correct management of insulinotherapy can be tabulated in accordance with different possible postoperative situations. PMID- 6287633 TI - [Control of blood glucose and insulin therapy in a surgical milieu]. PMID- 6287634 TI - [Screening for blood glucose abnormalities and coronary heart disease through routine annual examination (author's transl)]. AB - The authors address the two following questions: 1) Is detection of glycoregulation disorders possible from data yielded by routine annual examination (7,923 subjects)? 2) Do further investigations, carried out in a subdivision of subjects selected according to the risk of cardiovascular disease (n = 1,232), show a difference in the distribution of risk factors and risk indicators between patients and "healthy" individuals? Analysis of the results of this transversal study demonstrates a fact that is well-known concerning diabetes: even mild hyperglycemia (greater than or equal to 6 Mmol/l) is significantly correlated to age, excess weight and high systolic blood pressure. In both male and female subjects, there is a strong correlation between mean blood glucose concentration and the other biologic parameters that were studied (blood pressure, serum cholesterol, LDL/HDL cholesterol). Conversely, no correlation was found with smoking habits or with the intake of various dietary components. These results indicate that age, excess weight and increased blood pressure are simple data which are suggestive of abnormal glycoregulation. Comparison of risk factors and risk indicators in patients and "healthy" subjects showed significant differences for age, smoking habits and systolic blood pressure only. Glycemia, cholesterolemia and obesity were similar in both groups. These findings are evidence of the high risk in the subjects considered as "healthy", in whom the degree of atherosclerosis cannot be ascertained. Atherosclerosis in "healthy" individuals can certainly not be overlooked: in subjects with normal blood pressure, fundus examination showed a similar proportion of abnormalities in patients and "healthy" individuals. PMID- 6287635 TI - [Arteriopathy of the lower limbs and giant-cell arteritis. Diagnosis on arteriographic and histologic examinations (author's transl)]. AB - A case of giant-cell femoral arteritis seen in patient with polymyalgia rheumatica is reported. Diagnosis was ascertained by demonstrative arteriographic images and unequivocal features upon histologic examination of in situ samples. With reference to this report and to a review of the medical literature, nosologic problems in giant-cell arteries of large arteries are discussed, clinical features are reviewed and diagnostic criteria are assessed. PMID- 6287636 TI - [Skin flora of the hands, elbow-hollow and fore-arm (author's transl)]. AB - During five years, the values of the skin flora of the hands, fore-arm and elbow hollow were studied on volunteers, especially five of them. Samples were taken according to the washing method. Results show important quantitative variations of the flora between the differents volunteers and even for a same volunteer. In the five long term followed subjects, the differences were: 2.17 log 10 for aerobic flora and 1.97 log 10 for aeroanaerobic flora of the hands, 3.90 log 10 for aerobic flora and 3.88 log 10 for aero-anaerobic flora of the fore-arm, 2.95 log 10 for aerobic flora and 3.35 log 10 for aero-anaerobic flora of the elbow hollow. These variations remained independent of the season. According to these variations, already notified by others authors it is suggested first to standardize sampling and bacteriological methods, second to develop multicentric programs in view to increase the number of subjects to be included in the study. PMID- 6287637 TI - [Role of Chester-Erdheim disease in histiocytosis]. PMID- 6287638 TI - [Iatrogenic ergotism. Responsibility of an ergotamine tartrate-erythromycine propionate association (author's transl)]. AB - Acute ischemia of the lower limbs was seen in a patient given ergotamine tartrate with erythromycine propionate. The authors recall the danger of such associations in patients with hepatic failure, the value of early diagnosis, and the possible use of nitroglycerin for suppressing the vascular spasm. PMID- 6287639 TI - [Secondary neoplastic involvement of the hepatic pedicle (author's transl)]. AB - Neighboring tumors can extend to the hepatic pedicle, usually through lymphatic involvement. Four such cases are reported. Thrombosis of the portal vein may develop. Cholestatic jaundice is the main clinical feature. Prehepatic portal hypertension may be present. Hepatojejunostomy is often the only possible surgical procedure. PMID- 6287640 TI - [Leiomyosarcoma of inferior vena cava. A new case. Surgical management (author's transl)]. AB - The authors report on a new case of leiomyosarcoma of the inferior vena cava, to add to the sixty five cases already mentioned in the literature. The patient had been suffering from subcostal pains for 12 years, and an adenomyoma of the gall bladder was diagnosed. The tumor was discovered in the course of a cholecystectomy. The vena cava was resected and ligated below the renal veins. After describing the case, the authors go on to review the literature and stress the fact that the prognosis, which is often severe, could be improved by earlier diagnosis. PMID- 6287641 TI - [Report on two cases of agitation in respiratory failure (author's transl)]. PMID- 6287642 TI - [What's new apropos of breast prostheses?]. AB - What is new in the field of mammary prostheses? In answer to this question, the authors reply "Nothing". They stress the need for a new approach to the problem of capsular contraction based on: 1) experimental research aiming at identification of the collagen and the capsular fibroblasts, by cell culture, in particular; 2) the abandoning of horizontal in favour of longitudinal statistics based on prolonged multifactoral observation of the largest possible number of patients; 3) greater scientific rigour in the analysis of prosthetic insertion in augmentation mammoplasties and mammary reconstructions. In this connection, they describe their own technique for augmentation mammoplasties by insertion of an inflatable prosthesis placed retropectorally by an areolo-mamillary approach. PMID- 6287643 TI - [Hormonotherapy in acute, idiopathic hirsutism, and seborrheic alopecia (author's transl)]. AB - It has been established that, through skin responds to circulating androgens, it can also produce androgens from precursors. Therapy can address either of the two following physiopathological steps: the secretory step can be countered by replacement or suppressive therapy which is now fairly well mastered; the target organ step opens new possibilities concerning receptor abnormalities and receptor control. The medical literature was reviewed for data on hormonal therapy for women with acne, idiopathic hirsutism and seborrheic alopecia, three conditions which are usually only partially improved by cosmetic treatment. PMID- 6287644 TI - The hepatitis-B virus and hepatocellular carcinoma. PMID- 6287645 TI - Hepatic lesions caused by anabolic and contraceptive steroids. PMID- 6287646 TI - Pathology of alcoholic liver diseases. PMID- 6287647 TI - Malignant insulinoma. PMID- 6287649 TI - [Role of hormonal factors in the development of arterial hypertension in obesity]. PMID- 6287650 TI - Collagenolytic enzyme systems in human intervertebral disc: their control, mechanism, and their possible role in the initiation of biomechanical failure. AB - Changes in the macromolecular matrix of the intervertebral disc may predispose to biomechanical failure of the disc. Such changes would involve extracellular enzymes capable of altering the collagen and proteoglycan of the disc matrix. In this study, extracts from homogenates of both annuli fibrosi and nuclei pulposi of human lumbar discs were found to contain neutral collagenolytic, gelatinolytic, and elastinolytic enzymes in all samples assayed. The specific activity of the collagenase was higher in extracts from nuclear than from annular tissue, a difference not seen with gelatinase or elastase. The collagenase was present as both a latent and active enzyme, the other enzymes being fully active. Human intervertebral discs therefore contain enzymes capable of degrading their extracellular macromolecular matrix. PMID- 6287651 TI - Carcino-embryonic antigen in hepatocellular carcinoma. Report of a patient with very high serum concentrations. AB - Serum carcino-embryonic antigen (CEA) concentrations are frequently raised in patients with hepatocellular carcinoma, but the degree of elevation is slight or moderate. In contrast, patients with hepatic metastases from colonic carcinomas and various other entodermal and non-entodermal tumours have very high serum values. This difference may be useful in the differential diagnosis between primary and metastatic liver carcinomas. However, exceptions do occur, as shown by the present report of a patient with hepatocellular carcinoma who had a serum CEA concentration of 1,600 ng/ml. PMID- 6287648 TI - Indirect hemagglutination antibody titers for Entamoeba Histolytica in dried filter paper blood and sera. AB - Serum and elutates from dried filter paper bloods from 404 patients with and without amebic liver abscess (ALA) were tested for Entamoeba histolytica antibodies by the indirect hemagglutination test. Eighty-nine percent of the sera from patients with ALA were positive (titers greater than or equal to 1 : 128) and 94% of those without ALA negative. Fifty-five percent of the filter paper bloods from ALA patients were positive. Overall agreement was 80% for positives and negatives and the correlation coefficient between the methods showed a linear relationship (r = 0.77). Although the quantity of sera available from dried filter paper bloods is limited and some antibody activity may be lost, the method should be of value in seroepidemiologic studies where venous bloods are not easily obtained. PMID- 6287652 TI - Serum immunoreactive calcitonin concentrations in hepatocellular carcinoma. AB - Having found raised serum calcitonin concentrations in 94% of patients with hepatocellular carcinoma when using a dextran-coated-charcoal radio-immunoassay, we have now repeated the study, using a double-antibody radio-immunoassay, in 102 further patients with hepatocellular carcinoma and 35 matched controls. Serum immunoreactive calcitonin concentrations (iCT) in the controls ranged from 10 to 310 pg/ml (mean 154.6 pg/ml). Values in the tumour patients ranged from 10 to 1,650 pg/ml (mean 302.6 pg/ml). The mean figures were significantly higher in the tumour patients (P less than 0.001), 35.5% of them having values above 310 pg/ml. In 65 of the patients serum iCT concentrations were also determined by dextran coated-charcoal radio-immunoassay. Values ranged from 10 to 10,780 pg/ml (mean 2,179 pg/ml). If 1,000 pg/ml is taken as the upper limit of normal, 69% of the patients had raised iCT concentrations. There was a good correlation (r = 0.67; P less than 0.001) between serum iCT values measured with both methods in 50 patients. If measured by the double-antibody radio-immunoassay method, the serum calcitonin value is not useful as a marker for hepatocellular carcinoma. PMID- 6287653 TI - Susceptibility of dermatiaceous fungi to amphotericin B, miconazole, ketoconazole, flucytosine and rifampin alone and in combination. AB - The dematiaceous fungi comprise a group of organisms that are deeply pigmented and found in soil or on decaying organic material, such as wood. The majority of infections with these fungi presumably results from traumatic inoculation. Although various forms of infection have been appreciated for some time, none of the presently available antifungal drugs have been shown to have predictable activity against these organisms. We report on the activity in vitro of various antifungal agents alone and in combination against various dematiaceous fungi. PMID- 6287654 TI - [Relationship between oral tissues and human tumor viruses (author's transl)]. PMID- 6287655 TI - Genetics of receptors for bioactive polypeptides: expression of the human EGF receptor gene and internalization and processing of the receptor-bound EGF in human-mouse cell hybrids. AB - We previously postulated that the structural gene for epidermal growth factor (EGF) receptor is located on human chromosome 7 (1,2). In this study, EGF receptor and certain postreceptor functions were further analyzed in a unique cell hybrid line, C2B5, that retains only one human chromosome of an X;7 translocation besides a nearly complete mouse parental genome. Kinetics and Scatchard analysis of [125I]EGF binding to the C2B5 hybrid cells indicated that they carry a single class of EGF receptors with a dissociation constant of 4 x 10(-10) M. The receptors expressed in the hybrids are proven to be immunologically of human nature. The human EGF receptors now embedded in essentially mouse plasma membrane are subject to "down regulation" mediated by the ligand EGF. Analysis of the cell-bound EGF indicated that internalization and processing take place in the human-mouse cell hybrids. The degradation of EGF appears to be through a lysosomal pathway since it was substantially delayed or inhibited by lysosomotropic agents. PMID- 6287656 TI - A clinical study of cholangiocarcinoma caused cholestasis in Thailand. AB - Cholangiocarcinoma is the leading cause of cholestasis among patients with malignant tumors, 63.5 per cent, while carcinoma of the head of the pancreas is the most common cause in western countries. A clinical history of progressive cholestasis; signs and symptoms of cholestasis; slow progressive clinical course, and mostly importantly, 96.7 per cent stemming from an endemic area of opisthorchiasis are all shown in this study. The peak age for this disease in Thailand was about one decade younger than that in western countries. The anatomic lesions in 96.7 per cent were found in the hilar area of the liver and only 3.3 per cent in the lower third of the liver compared with a significant number of lesions in the middle and lower thirds of the liver, as reported in literature from the western countries. We believe that percutaneous transhepatic cholangiography prior to the operation will definitely help the surgeon in deciding upon the surgical approach. Surgical intervention of a cholangiocarcinomatous lesion is probably more difficult in Thailand than in western countries because of location. The incidence for cholangiocarcinoma associated with biliary calculi and for an opisthorchiatic cyst is 8.1 and 6.1 per cent. We would like to emphasize the close association between cholangiocarcinoma and the combination of opisthorchiasis with carcinogenic agents, such as nitrosamines in the favorite local dish. The animal experimental model, using Syrian golden hamsters, produced the same result and the same site for the lesion--the hilar area of the liver. PMID- 6287657 TI - Action of cholecystokinin-octapeptide on sphincter of Oddi basal pressure and phasic wave activity in humans. AB - The human sphincter of Oddi (SO) exhibits phasic wave activity over a 4 to 6 mm segment. Approximately 60% of these waves occur in an antegrade direction, 14% are retrograde, and 26% occur simultaneously. Because cholecystokinin-octapeptide (CCK-OP) stimulates the flow of bile into the duodenum, its effect on SO phasic wave contractions and contraction sequences was evaluated at ERCP manometry. An infused triple-lumen catheter of 1.7 mm outer diameter with side orifices spaced 2 mm apart was stationed in the SO segment so that all three orifices recorded phasic pressure waves. We studied 31 patients with normal ductal anatomy and normal SO pressures. In 21 of these patients CCK-OP (20 ng/kg) was given intravenously after a 2- to 3-minute baseline recording was obtained. Pressure recordings were continued for up to 10 minutes following CCK-OP administration. CCK-OP caused a significant inhibition in the frequency and amplitude of SO phasic waves as well as a significant decrease in basal SO pressure. Before CCK OP most phasic contractions were antegrade, and after CCK-OP the sequence pattern remained unchanged. We conclude that CCK-OP reduces or transiently abolishes SO phasic contractions but that it does not change their temporal sequence. In addition, CCK-OP produces a decrease in basal SO pressure. These findings suggest that the action of CCK-OP in humans is to inhibit SO phasic activity and reduce SO basal pressure to allow increased flow of bile. PMID- 6287658 TI - Immune modulatory effects of indomethacin in melanoma patients are not related to prostaglandin E2-mediated suppression. AB - Indomethacin significantly enhances the depressed levels of lymphocyte proliferation to the mitogens phytohemagglutinin and concanavalin A in melanoma patients. We postulated that these results were related to an abnormality in prostaglandin E2 (PGE2)-mediated suppression, since this mechanism has previously has previously been demonstrated in patients with Hodgkin's lymphoma and with head and neck carcinoma. However, the results of three experimental approaches did not support this hypothesis. First, in vitro PGE2 production by cultured blood mononuclear cells was the same in 16 melanoma patients as in 45 normal controls (4.9 versus 4.7 ng/ml). Second, lymphocyte sensitivity to PGE2 for melanoma patients was essentially the same as that for normal controls, since exogenous doses of PGE2 inhibited the mitogen responses to the same degree. Third, another prostaglandin synthetase inhibitor (RO-205720), which is structurally unrelated to indomethacin, did not augment the mitogen response in these patients. Thus PGE2 cannot be implicated as a mediator of immunosuppression in melanoma patients. To further examine the immunomodulatory mechanism of indomethacin, we preincubated the drug with purified populations of either lymphocytes or monocytes, which were then recombined and tested for mitogen response. The results suggested that indomethacin had a direct effect on the responding T lymphocytes rather than an indirect effect on monocytes. These are the first studies demonstrating that indomethacin can act directly as a modulator of cellular immune function, independent of PGE2-mediated suppression. PMID- 6287659 TI - [Hypothalamo-hypophyseal-adrenal system function in rheumatoid arthritis]. PMID- 6287660 TI - Respiratory cytodiagnosis: study in observer variation and its relation to quality of material. AB - Sensitivity and accuracy of cytodiagnosis were assessed in a multicentre study. Six centres each provided sputum cytological material from 20 cases. Each centre screened and reported on the 100 slides provided by the other five centres. The reports were assessed against consensus reference diagnoses, reached by discussion with transparencies, histological sections, and closed-circuit television. False positive rates of 0-4% (average 1.3%) and false negative rates of 0-12% (average 5.0%) of slides examined were recorded. The order of agreement on the three common cell types was adenocarcinoma 75% (50-91%), squamous cell carcinoma 80% (59-94%) and small carcinoma 95% (71-100%). The effect of quality of material on cytological opinion was assessed by comparing disagreement rates on each of the different sets of 20 slides. Disagreement varied from 1% to 23% depending on which set of material was examined. PMID- 6287661 TI - Coexisting granular cell myoblastoma and squamous carcinoma of the bronchus. PMID- 6287662 TI - Subcellular localization of adenosine diphosphatase in cultured pig arterial endothelial cells. PMID- 6287663 TI - Phosphoinositide metabolism in human blood platelets: effects of two types of divalent cation chelators. AB - Phospholipid metabolism was investigated in platelets in relation to secretion of 5-hydroxytryptamine. Both thrombin and the ionophore, A23187, induced secretion of 5-hydroxytryptamine and platelet aggregation accompanied by the specific breakdown of phosphatidyl inositol. The products detected corresponded to phosphatidic acid and a substance which co-chromatographed with polyphosphoinositides and lysophosphatidyl inositol. Thrombin-induced aggregation, secretion and phosphatidyl inositol metabolism were all prevented by EDTA, but only the secretory response was inhibited by the intracellular calcium chelator chlortetracycline (CTC). In contrast, A23187 induced turnover of phosphatidyl inositol in the presence of EDTA even though aggregation and secretion were prevented. All three responses to A23187 were inhibited by chlortetracycline. Thus, platelet aggregation, secretion and phospholipid metabolism can be dissociated by the use of two types of divalent cation chelators: EDTA and CTC. Furthermore, these findings suggest that thrombin and A23187 induce phosphoinositide metabolism by two distinct mechanisms. PMID- 6287664 TI - Role of macromolecular binding site of thrombin molecule in excitation of anticoagulation system. AB - The reaction of anticoagulation system upon perfusion of humorally isolated (with retained innervation) carotid sinus of a rabbit by alpha-,beta/gamma-, DIP-alpha thrombin and prethrombin I was studied. DIP-alpha-thrombin without clotting activity was shown to initiate like alpha-thrombin the reflex reaction of anticoagulation system characterized by a sharp increase in non-enzymatic fibrinolysis (by 225%) and total fibrinolytic activity of blood (by 51%). Prethrombin I (thrombin precursor) is also capable of exciting the function of anticoagulation system characterized by an increase in non-enzymatic fibrinolysis (by 82%) and total fibrinolytic activity (by 36%). Furthermore, perfusion of prethrombin I or alpha-thrombin at almost the same molar concentrations resulted in the similar degree of anticoagulation system effector reaction. Reflex response of anticoagulation system was not observed upon perfusion of carotid sinus by beta/gamma-thrombin that has high esterase but little if any clotting activity that appears to be due to molecular changes in the macromolecular binding site region. These data support the suggestion that the effect of anticoagulation system excitation is due to interaction of the macromolecular binding site in the structure of alpha-thrombin with anticoagulation system chemoreceptors. PMID- 6287665 TI - Effects of galactose-binding lectins on human blood platelets: identity of the peanut agglutinin receptor with the von Willebrand factor receptor. PMID- 6287666 TI - [Aujezky's disease in sheep caused by errors in vaccines]. PMID- 6287667 TI - ACTH content in the anterior pituitary after the infusion of various doses of corticosterone and dexamethasone in normal and adrenalectomized rats. AB - It is well known that the administration of glucocorticoids inhibits the activities of hypothalamus-pituitary-adrenal axis. The present studies were undertaken to define the site of inhibition of both corticosterone and dexamethasone on this axis in rats. Infusion of 20 or 202 micrograms corticosterone over 6 hr did not change the ACTH content in the anterior pituitary and plasma ACTH levels even through the infusion markedly inhibited the hypothalamic CRF content in normal rats. The constant infusion of a relatively small dose of 20 micrograms dexamethasone inhibited the plasma ACTH level, but not the ACTH content in the anterior pituitary and the hypothalamic CRF content. In contrast to the above results, the infusion of 202 and 504 micrograms dexamethasone over 6 hr increased the ACTH contents in the anterior pituitary, but significantly reduced the hypothalamic CRF content and plasma ACTH levels. These results suggest that the mechanism of dexamethasone inhibition in the hypothalamus-pituitary-adrenal axis is the reduction of ACTH release rather than that of ACTH production in the anterior pituitary. PMID- 6287668 TI - A case of early gastric cancer whose diagnosis was expedited by amogastrin-99mTc pertechnetate scintigraphy. PMID- 6287669 TI - Frequencies of specific antibody-producing cells for sheep red blood cells and for phosphorylcholine in human tonsillar B lymphocytes transformed by Epstein Barr virus. AB - The frequencies of transformed B cells and of transformed specific antibody producing cells in human tonsillar lymphocytes infected with Epstein-Barr virus (EBV) were determined by limiting dilution analyses. The latter was corrected by the former and the frequencies of specific antibody-producing cells in EBV transformed B cell populations were obtained. The frequencies of specific B cells for sheep red blood cells were calculated to be one in 500 to 2,000 EBV transformed tonsillar B cells obtained from four donors, while those of specific B cells for a hapten, phosphorylcholine, were one in 700 to 5,000. The method may be useful to define the repertoire of specificities in the human B cell populations. PMID- 6287670 TI - A correlation between cancer metastases and the fluidity of cancer cell membrane. AB - Male Donryu rats weighing from 100 to 120 g were transplanted with AH100B, AH66F or Yoshida sarcoma (YS) cells intraperitoneally. After collecting ascites, while not bloody, the cancer cells were washed three times with Dulbecco's phosphate buffered saline (PBS) without Ca++ and Mg++ (pH 7.2). Then, each sediment was suspended in PBS mentioned above. Each suspension in which 10 million of cancer cells were contained was violently vibrated with a definite amount of 5-doxyl stearic acid and spin labeling of cancer cell membrane was done. Furthermore, each specimen thus obtained was subjected to measurements of ESR spectra and the order parameter was determined from the spectra. The order parameter of AH100B was the highest and that of YS the lowest at each temperature measured from 5 degrees C to 35 degrees C. YS cells have been reported to be found in blood stream within 24 hr, AH66F cells in a day and AH100B cells in 10 days or more after intraperitoneal inoculation. The results obtained here suggest that the accelerated fluidity of cancer cell membrane is related with the cancer metastasis. PMID- 6287671 TI - Comparison of cytosolic and mitochondrial enzyme alterations in the livers of propionic or methylmalonic acidemia: a reduction of cytochrome oxidase activity. AB - The activities of mitochondrial, cytosolic and microsomal enzymes in liver specimens obtained from three patients with propionic or methylmalonic acidemia were compared with those of control patients who had died from unrelated causes. Only the activity of cytochrome oxidase (mitochondrial enzyme) was significantly reduced in the patients of propionic acidemia and methylmalonic acidemia who were in the state of metabolic acidosis; in two patients the activity was less than 30% of that in controls, but in the other patient of propionic acidemia, who was under the treatment with a low protein diet (0.8 g/kg/day), the activity was 50% of that in controls. The metabolites of branched chain amino acids (tiglic acid, propionic acid, methylmalonic acid, succinic acid, tiglyl-CoA and propionyl CoA) exhibited no inhibitory effect on the cytochrome oxidase activity of the sonicated rat liver mitochondria. The reduction of cytochrome oxidase activity found in these organic acidemias may be caused secondarily by some unknown mechanism. PMID- 6287672 TI - Spanish adulterated oil matter. An important discovery by Spanish toxicologists: the toxicity of anilides of unsaturated fatty acids. AB - Collective intoxication resulting from the addition of aniline in adulterated vegetal oils which were re-treated in order to make them available for consumption, was clinically characterized by polyneuritis, atypical pneumonia, symptoms of vascularity. After studying the infection and allergic syndrome, the toxicological approach presented a set of experimental clinical arguments in favor of a specific toxicity of anilides of unsaturated fatty acids. The toxicokinetics of oleylaniline shows a half life of several weeks, which explains its evolution. Its mode of action involves peroxide formation, membrane toxic and attractive substances of immunocompetent cells. Peroxide formation could result from an excess of superoxide ions, an inhibition of superoxide-dismutases, an excessive consumption of NADPH or more often of any facilitation of lipid peroxidation by a biologically active peroxidized intermediate. PMID- 6287673 TI - Acnegenic activity of 3-methylcholanthrene and benzo[a]pyrene, and a comparative study with 2,3,7,8-tetrachlorodibenzo-p-dioxin in the rabbit and hairless mouse. AB - The non-halogenated hydrocarbons 3-methylcholanthrene (MC) and benzo[a]pyrene (BP) were tested for acnegenic activity using the rabbit ear test. Both compounds induced characteristics follicular hyperkeratosis, although their acnegenic potency was approximately 4 orders of magnitude lower when compared to the potent acnegen 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). These results are discussed in view of the hypothesis that the acnegenic activity of TCDD and its congeners is mediated through stereospecific binding to a cytosolic receptor protein. In an experiment with hairless (hrhr) mice, which mutant has been described as an animal model for testing acnegenic potency, reduction in sebaceous gland tissue but no follicular hyperkeratosis was observed after application of a total dose of 0.4 microgram TCDD on the back skin. At a 10 times lower dose, no effects were seen. The hairless mouse strain used was responsive to TCDD as judged from the dose-related increase in the activity of aryl hydrocarbon hydroxylase (AHH) in liver microsomes, the increased liver weight and the histopathological changes in the liver. In comparison, a total dose of 0.12 microgram TCDD produced a strong follicular hyperkeratosis in the rabbit ear. From these results and from literature data, the adequacy of the hairless mouse for the testing of compounds for acnegenic potency is questioned. PMID- 6287674 TI - Biochemical response of rat lung to inhaled n-hexane. AB - Biochemical response of the rat lung to inhaled n-hexane was investigated. Dose dependent increase in protein, lipid, sialic acid, lactate dehydrogenase, angiotensin-converting enzyme, acid and alkaline phosphatase was observed in the cell-free lavage effluent of the lungs of exposed rats compared to the control animals. This was interpreted as enhanced pulmonary secretions accompanied by increased cell damage. PMID- 6287675 TI - Differences in benzo(a)pyrene metabolism between cultured human and murine bronchial cells after pre-treatment with benz(a)anthracene. AB - Primary cultures of human and murine (strain C3Hz) bronchial epithelial cells were pretreated with benz(a)anthracene (BA) (10 microM). 16 h later the formation of phenolic as well as dihydrodiol metabolites of benzo(a)pyrene (BP) was measured. Whereas murine cultures showed enhanced metabolism towards both phenolic and dihydrodiol compounds, in the human cultures only phenolic BP metabolites were increased. In view of their precursor role in the formation of biologically active diol-epoxides, formation of dihydrodiol-derivatives can be considered as a key factor in determining susceptibility to polycyclic aromatic hydrocarbon (PAH)-induced carcinogenesis. Therefore the observations of this study indicate that animal model systems for PAH carcinogenesis in man have to be selected on the basis of comparable metabolite patterns. PMID- 6287676 TI - Comparative toxicities of aliphatic nitriles. AB - Aliphatic nitriles have been postulated to manifest their toxicity through cyanide liberation. We have investigated the signs of toxicity and effect of equitoxic LD50 doses of saturated and unsaturated aliphatic mono- and dinitriles on tissue and blood cyanide levels, tissue glutathione levels and cytochrome c oxidase activities. Signs of toxicity were classified into cholinomimetic effects observed with unsaturated nitriles and central nervous system effects observed with saturated nitriles and potassium cyanide (KCN). Hepatic and blood cyanide levels 1 h after treatment were highest following malononitrile (MCN) and decreased in the order of propionitrile (PCN) greater than KCN greater than butyronitrile greater than acrylonitrile (VCN) greater than allylcyanide greater than greater than fumaronitrile greater than acetonitrile. The order differed in brain where KCN preceded MCN and PCN. Hepatic and brain cytochrome c oxidase were significantly inhibited and corresponded to their cyanide levels. No significant inhibition of cytochrome c oxidase was observed in vitro. VCN was the only nitrile which significantly reduced tissue GSH levels. In conclusion, toxic expression of aliphatic nitriles depends not only upon cyanide release but also on their degree of unsaturation. With unsaturated aliphatic nitriles cyanide release plays a minimal role in their toxicity. PMID- 6287677 TI - TCDD embryotoxicity in the mouse may be enhanced by beta-naphthoflavone, another ligand of the Ah-receptor. AB - The significance of the aryl hydrocarbon (Ah)-receptor (which binds 2,3,7,8 tetrachlorodibenzo-p-dioxin (TCDD) and a number of polycyclic hydrocarbons) in murine embryonic tissues for the teratogenic action of TCDD has been studied. The receptor has been localized, especially to the embryonic maxillary region. When the non-teratogenic beta-naphthoflavone (beta-N), which is another ligand of the Ah-receptor, was co-administered with TCDD it increased the frequency of cleft palate compared with the number induced by TCDD alone. This was true when beta-N was administered simultaneously or 8 h before TCDD but not 24 h before or after. Fetal death was induced even when the two compounds were administered at wider intervals. It is suggested that TCDD and beta-N partly induce a battery of enzyme systems leading to the final malformation. PMID- 6287678 TI - [Phosphatase activity of the neutrophilic leukocytes in acute odontogenic inflammation of the jaws]. PMID- 6287679 TI - [Therapy result of malignant tumors of the paranasal sinuses in patients treated at the metropolitan onco-radiologic centre of Budapest. Period under review: 1946 1974 (author's transl)]. AB - A report is given on the therapy results of 301 patients with ethmoidomaxillary tumors treated at the Metropolitan Onco-Radiologic Centre of Budapest between 1946 und 1974. Most of these patients were in an advanced stage when the first diagnosis was established. The five-year survival rate of all stages was 27%, which does not strongly differ from the average value of 30% mentioned in literature. The five-year survival rate of the patients submitted to a combined surgical and radiologic treatment was 33%, that of the patients treated by irradiations only was 16%. 81 patients received an intracavitary brachycurietherapy after having undergone a non-radical operation. The five-year survival rate of this group was 38%. The intracavitary brachycurietherapy is most favorable and should always be applied in case of this not very radiosensible, often even radioresistant tumor. PMID- 6287681 TI - The surgery of deep orbital tumors. PMID- 6287682 TI - The pectoralis major myocutaneous flap: its use in reconstruction following head and neck surgery. PMID- 6287680 TI - Activities of purine nucleotide metabolizing enzymes in human thymocytes and peripheral blood T lymphocytes: in vitro effect of thymic hormones. AB - Adenosine deaminase (ADA, E.C. 3.5.4.4) and purine nucleoside phosphorylase (PNP, E.C. 2.4.2.1) activities are essential for the normal development and function of T lymphocytes. A comparison of the specific activity of these two enzymes and of ecto-5'-nucleotidase (5'-N, E.C. 3.1.3.5) of human thymocytes with that of peripheral T lymphocytes of children and young adults shows that significant differences exist between the activities of ADA and 5'-N in thymocytes and peripheral T cells. ADA activity is six-fold higher in thymocytes than in peripheral T cells whereas 5'-N activity is approximately one-fourth lower in thymocytes than in T lymphocytes. PNP activity is two-fold higher in T cells. The apparent Km and Vmax values for the three enzymes in thymocytes and peripheral T lymphocytes have been determined. The observed differences in enzyme activity are probably not entirely due to differences in the affinity of the enzymes for their substrates. The enzyme activities of a thymoma removed from a patient with myasthenia gravis had intermediate enzyme levels for ADA and PNP, but 5'-N was similar to peripheral T cells. Exposure of thymocytes in short-term culture to the thymic hormones thymopoietin (the pentapeptide TP5), thymosin fraction V or serum thymic factor (FTS) had no significant stimulatory or inhibitory effect on the activities of the three enzymes under our experimental conditions. PMID- 6287683 TI - Circulating factor from a kala-azar patient suppresses in vitro antileishmanial T cell proliferation. AB - In vitro lymphocyte blastogenesis stimulated by phytohaemagglutinin (PHA), streptococcal antigens (SKSD) and leishmanial antigens were used to assess T cell responsiveness in one patient with kala-azar before and after therapy. During the illness, responses to PHA and SKSD but not to leishmanial antigens could be detected. After treatment lymphocytes responded to all three stimuli when cells were cultured in convalescent plasma, but failed to respond to leishmanial antigens when cultured in plasma obtained before treatment. These observations suggest the presence of a circulating inhibitor of anti-leishmanial T cell responsiveness in kala-azar, and warrant further investigation. PMID- 6287684 TI - High incidence of IgG class of Epstein-Barr virus capsid antibody in Indian patients of lepromatous leprosy. AB - Low levels of Epstein-Barr virus capsid (EBVC) antibody of the IgG class were detected in the sera of 19 of 23 (82.6%) lepromatous leprosy patients and six of 38 (16%) healthy controls. In contrast, heterophile antibody was found in only six of 43 (14%) lepromatous patients and three of 41 (7%) normal subjects. Overlap of the two types of antibody occurred only in one normal serum. It is inferred that the presence of EBVC antibody against an unbiquitous virus in lepromatous patients who often suffer from impairment of cell-mediated immunity might be due to past infection leading to persistence of the virus in their lymphoid cells and subsequent production of specific anti-viral antibody. Further, the striking finding of low incidence and titre of EBVC antibody in the normal Indian adults is consistent with the rarity of EB virus-associated disease, such as Burkitt's lymphoma, nasopharyngeal carcinoma and infectious mononucleosis in India. PMID- 6287685 TI - Preliminary observations using a multi-layer ELISA method for the detection of Entamoeba histolytica trophozoite antigens in stool samples. AB - A method for detecting Entamoeba histolytica trophozoite antigens in aqueous solution is described. This involves coating plastic microtitre plates with a 'catching antibody', specific rabbit anti-E. histolytica antibody (SRAE). After adding the test material the presence of antigen is determined using two additional heterologous antibody layers, one of 'developing antibody', in this case human anti-E. histolytica immunoglobulin (HAE), which is followed by a final layer of peroxidase conjugated sheep anti-human immunoglobulin antibody (SH-HRP). The specificity and sensitivity of the assay was investigated both in the model system and using stool samples from infected patients. In the model system, the test had a sensitivity equivalent to detection of approximately one amoeba per microscope coverslip (18 mm X 18 mm). Little cross reaction was observed with other intestinal parasites common to the area of Bangladesh from which the stool samples were taken. The possible use of this method in large scale screening of stool samples and in the detection of circulating antigens is discussed. PMID- 6287686 TI - A zymodeme study of Entamoeba histolytica in a group of South African schoolchildren. AB - Using a biphasic culture medium, stocks of intestinal amoebae were isolated from a group of children attending school in Durban, South Africa. These were compared with stocks collected in other areas of the world already characterized using the electrophoretic patterns of four enzymes: glucose phosphate isomerase (GPI), phosphoglucomutase (PGM) L-malate: NADP+ oxido-reductase (oxalacetate decarboxylating) (ME) and hexokinase (HK). 33% of 94 samples grew Entamoeba histolytica, only one of which gave a pattern indicative of a pathogenic stock. Entamoeba hartmanni, Dientamoeba fragilis and Entamoeba coli were also grown from some samples, increasing the total positive samples for all species isolated to 40%. PMID- 6287688 TI - [Observations on tsetse flies in a forest focus of human trypanosomiasis in Ivory Coast. 3. Dispersal and distribution of fly populations around a village (author's transl)]. AB - In the Vavoua human trypanosomiasis focus (forest zone of Ivory Coast) four tsetse fly catching series, of nine days each, were made during the rainy season using eighty five biconical traps. The dispersion of flies in the study area was studied by the mark-release-recapture method. Two series of two sets of capture mark-release were carried out. In the first set flies were captured, marked and released in their point of origin and in the second set captured and marked in the village periphery but released in coffee plantations and vice versa. A total of 10198 tsetse flies were marked and released and 267 were recaptured. There was no difference between recapture rates of the different village or plantation released G. palpalis; this suggested that the same population was living in different biotops. Males and females from the periphery of the village behaved differently. Males dispersed evenly in contrast to females which remained in their original area. This difference in dispersion of the two sexes may be of epidmiological importance for human trypanosomiasis. Of interest was the discovery of a small community of peri-domestic G. pallicera during the periods March--April and May. The estimation of tsetse population by the Lincoln Index, is related to the dispersive power of the species concerned. When evaluated through the recapture rate this was found to be ten and seven times higher for G. pallicera and G. nigrofusca respectively, than for G. palpalis. This difference may be important in explaining the phenomenon of reinvasion and the little apparent effect of insecticidal control on the two former species. PMID- 6287687 TI - Epidemiological studies on the animal reservoir of Gambiense sleeping sickness. Part III. Characterization of trypanozoon stocks by isoenzymes and sensitivity to human serum. AB - Polymorphism in 12 enzymes, as shown by electrophoresis on thin-layer starch-gel, was examined in 88 stocks of trypanosomes of the subgenus Trypanozoon isolated from man and animals in the Ivory Coast and Upper Volta. Three of the enzyme profiles seen in trypanosomes from man in the Ivory Coast were exactly the same as in trypanosomes from local domestic pigs and from various game animals and a bovine in the Upper volta, thus confirming previous evidence that human trypanosomiasis is a zoonosis in West Africa. Altogether 9 zymodemes were found in man; one was exactly the same as another from the Congo while a further one was identical to a Ugandan zymodeme. Thirty-one zymodemes were found only in animals, and 6 were exactly the same as others from elsewhere in Africa, including the eastern part. All zymodemes resembled each other by possessing common electrophoretic patterns in 5 enzymes. In most zymodemes, the variants of two other enzymes were characteristically West African, although an East African influence was apparent, together with further evidence of hybridization. Many zymodemes differed from others only to a minor extent in a few isoenzyme bands. A group of closely related minor zymodemes constituted another trypanosome population ineffective to man in West Africa which had a variable sensitivity to normal human serum; enzymatically it was clearly separated from the major zymodeme previously described in West Africa, which was consistently resistant to normal human serum and had been previously associated with chronic gambiense sleeping sickness. PMID- 6287689 TI - Evaluation of renal responsiveness to exogenous parathyroid hormone on the first and third days of life. PMID- 6287691 TI - Cystic nephroma: morphologic spectrum and implications. AB - Among 216 primary renal tumors collected from a pediatric population in metropolitan Chicago, 6 instances corresponded in multiloculated cystic masses conforming to the diagnostic criteria of so-called cystic nephroma. Histopathologically, the component tissues were entirely mature in 2 cases, but in the remaining 4 the septa between cysts contained variable amounts of primitive or embryonic tissues; in one of these, a grossly visible mass formed a nodule of typical Wilms tumor. Our experience agrees with previous reports that favor closer structural similarity of these lesions to nephroblastoma than to any known form of cystic malformation. This concept has important consequences on the therapeutic approach to preoperatively diagnosed multilocular cystic masses of the kidney, and supports the idea that nephrectomy is the treatment of choice. Metastases or local recurrence have not been reported in cases of cystic nephroma. PMID- 6287690 TI - Characteristic echographic features of circumscribed cancer. AB - Of 964 amputated breast cancers which had been examined preoperatively by ultrasound and mammography, 147 breast (17.0%) contained macroscopically circumscribed tumors and 596 cases (69.0%) showed infiltrative tumors on examination of the cut surface. It is well known that infiltrative types of cancer show ultrasonic images that usually possess clear cut signs of malignancy, such as jagged augmented boundary echoes and acoustic shadows. On the other hand, circumscribed types of cancer do not usually show such signs, but exhibit lobulated shapes and strong, coarse and non-uniform internal echoes. These latter features are intermediate between those of infiltrative cancers and those of solid benign tumors. As such circumscribed cancers are often difficult to diagnose correctly by ultrasonic techniques. The diagnostic accuracy rates by ultrasound were 88.0% in the infiltrative types and 69.4% in the circumscribed types. The ultrasonic pictures of the 147 cases of such circumscribed tumors are classified and discussed. PMID- 6287693 TI - Pitfalls in diagnosis of exophytic renal tumors. PMID- 6287692 TI - Mucinous adenocarcinoma of renal pelvis. PMID- 6287694 TI - Equine influenza infections in Great Britain, 1979. PMID- 6287696 TI - Bovine virus diarrhoea in Egypt. PMID- 6287695 TI - Papillomaviruses, papillomas and squamous cell carcinomas in sheep. PMID- 6287697 TI - Effects of various antibiotics on the control of bacteria in boar semen. AB - Nine antibiotics were tested for the control of 11 bacterial genera isolated from boar semen. Of the antibiotics tested, the aminoglycosides dibekacin, amikacin and gentamicin proved fairly effective. Minimum inhibitory concentrations of those antibiotics were low (less than 6.25 micrograms/ml) compared to those of antibiotics commonly used for semen diluents. Seventy-four semen samples were stored at 15 degrees C by means of dialysis in the presence of dibekacin (100 micrograms/ml). After seven days' storage, about 80 per cent of the samples showed no bacterial growth and the mean values for sperm motility an normal acrosomes were 75.4 per cent and 82.7 per cent respectively. Twenty sows which were inseminated with semen stored for up to 14 days produced 16 litters and the average litter size was 9.9. PMID- 6287698 TI - Intranasal vaccination of pigs against Aujeszky's disease. 1. Comparison of intranasal and parenteral vaccination with an attenuated vaccine in 12-week-old pigs from immunized dams. PMID- 6287699 TI - Dual adenovirus and distemper virus pneumonia in a dog. AB - A case of simultaneous infection of the lungs of a dog with canine distemper virus (CDV) and canine adenovirus (CAV) is described. The techniques employed are histological stains, immunoperoxidase technique, and transmission electron microscopy. Two viruses are shown to infect the same cells. The significance of dual infection of the same cells is discussed in comparison with literature data on in vitro experiments. PMID- 6287700 TI - Comparisons between two isolates of stomatitis papulosa virus. AB - The cytopathology and length of latency in single-step growth curves of two isolates of stomatitis papulosa virus are compared in this report. Isolate 721 was obtained from a calf with oral ulcers and isolate 8665 was obtained from a calf with respiratory disease and oral ulcers. In single-step growth curves, the latency period of isolate 721 was 8 h while that of isolate 8665 was 6 h. The cytopathic effect produced by isolate 721 in bovine lung cells was characterized by enlargement of the cell, cell-to-cell adherence and large intracytoplasmic accumulations of viral inclusion material. Isolate 8665 caused rapid cell degeneration and detachment, with small accumulation of viral inclusion material. Neither of the two strains grew in bovine alveolar macrophage cultures or in the respiratory epithelium of fetal bovine tracheal explants. Intragingival inoculation of these isolates in cattle resulted in oral lesions without clinical signs of respiratory of systemic involvement. Virus was recovered from the oral lesions and from nasal secretions for as long as 10 days. Inoculation of dexamethasone-treated cattle resulted in a similar clinical condition although virus was recovered for 20 days from oral lesions and nasal secretions. Seroconversions from negative to 1 : 2560 were detected in inoculated cattle by indirect immunofluorescence. PMID- 6287701 TI - Foot-and-mouth disease virus: immunogenicity and structure of fragments derived from capsid protein VP and of virus containing cleaved VP. AB - Peptide fragments were obtained from the immunogenic capsid protein VP3, ca. 24 kilodaltons (kd), of foot-and-mouth disease virus type A12 119ab by three procedures: (1) spontaneous proteolysis of in virion VP3 in tissue cultures to produce a 15 kd peptide, designated S fragment; (2) trypsin treatment of purified virus to produce a 16 kg peptide, designated T fragment; and (3) cyanogen bromide cleavage of purified VP3 to produce a 13 kd fragment. Following isolation and purification by gel electrophoresis, VP3 and each of the three fragments were immunogenic for livestock. Lyophilization appeared to impair the immunogenicity of VP3. In addition, viruses containing VP3 fragments produced either by the spontaneous- or trypsin-induced proteolysis were as immunogenic as virus with its VP3 intact. Amino acid sequencing of N-terminal regions revealed that the S fragment was homologous with the N-terminus of VP3, whereas the 13 kd fragment possessed a unique N-terminus. Thus, putative common immunogenic amino acid sequences would appear to reside within an overlap region of the 15 kd S and 13 kd fragments. Sequencing of cDNA prepared to viral genome RNA provided three kinds of information: it (1) placed the above overlap region in the second and third quarters of VP3; (2) demonstrated that the codons for the C-terminus of VP1 and N-terminus of VP3 are contiguous; and (3) supported earlier evidence that these same codons program a chain reversal where VP1 and VP3 are joined in the precursor polyprotein. PMID- 6287702 TI - Difference in protective immunity of the tongue and feet of guinea pigs vaccinated with foot-and-mouth disease virus type A12 following intradermolingual and footpad challenge. PMID- 6287703 TI - Bluetongue virus serotypes 20 and 17 infections in sheep: comparison of clinical and serological responses. AB - The clinical, virological and serological responses of sheep infected with an Australian bluetongue virus (BTV) isolate (serotype 20) were compared to responses in sheep inoculated with an American bluetongue isolate (serotype 17) with which it had shown cross-reactions in serum neutralization tests. In sheep inoculated with BTV 20, clinical signs were very mild and viremia was first detected by day 5; virus was isolated intermittently for a further 2 to 3 days. Neutralizing and precipitating antibodies were first detected in the serum of the sheep between 2 to 3 weeks following inoculation. In contrast, sheep inoculated with BTV 17 showed pyrexia and severe hyperemia of the nasolabial area and oral mucosa from day 7 to 17. Viremia was first detected on day 3 and extended to day 20, while the appearance and titers of serum antibodies was similar in both groups. After challenge with BTV 17 the sheep in both groups remained clinically normal, and virus was not detected in the blood; however, serum neutralizing antibody titers to both viruses increased 2 weeks after challenge and the mean titer of the two groups ranged from 1:250 to 1:640. PMID- 6287704 TI - Recovery of dual serotypes of bluetongue virus from infected sheep and cattle. AB - Dual serotypes of bluetongue virus (BTV) were recovered from field-collected samples of sheep and cattle blood. Two sheep, each infected with both BTV serotypes 10 and 17, were found in a flock with bluetongue disease associated with these two serotypes. One sheep infected with BTV serotypes 11 and 17 was found in a second flock; it was the only viremic sheep detected and was clinically ill. Dual serotype infections of one beef and two dairy cattle were found in three geographically separate herds: mixtures recovered were of BTV serotypes 10 and 17 and serotypes 11 and 17. Clinical signs of illness were absent in the cattle in two herds, but severe conjunctivitis was seen in several cows in a third herd, including the cow with a dual serotype infection (BTV 11 and 17). Two of the cattle with dual infections had no serological evidence of BTV as determined by the agar gel precipitin test; serum was not available from the other cow with a dual serotype infection. The significance of dual infections and immune tolerance are discussed. PMID- 6287705 TI - An indirect enzyme-linked immunosorbent assay (ELISA) for the detection of antibodies to maedi-visna virus. AB - An indirect enzyme-linked immunosorbent assay (ELISA) for the detection of antibodies to maedi-visna virus (mvv) in sheep is described, in which microtitre plates are sensitized with a partly purified preparation of mvv. The antibodies bound are detected by a horseradish peroxidase conjugate. The results obtained with ELISA on a total of 493 serum samples from several commercial flocks were compared to those of a routine agar gel precipitation test (AGPT) and a complement fixation test (CFT). All samples which scored positive in AGPT, CFT or both (20.8%) were also found positive by ELISA. In addition, with ELISA a further 11.5% of the samples were positive. Serum samples from maedi-free flocks, from sheep suffering from sheep pulmonary adenomatosis and from lambs immunized against other viruses were all negative by ELISA. The assay has been used routinely for some years and proved to be specific, sensitive and suited for screening of large numbers of serum samples. PMID- 6287706 TI - Inactivated bluetongue virus vaccine in lambs: differential serological responses related to breed. AB - A mixed breed flock of lambs, consisting of Suffolks, Hampshires, Columbias and Finnish breeds, were vaccinated with binary ethylenimine inactivated blue-tongue virus (BTV) serotypes 11, 17 and a mixture of 11 and 17 in aluminum hydroxide. Agar gel precipitin antibodies were used as an indicator of immunity. Sero conversion of Hampshires and Suffolk lambs was poor at 43% as compared to 84% in the Columbia and Finn lambs. These results indicate a breed difference in immunological response to inactivated BTV vaccine. PMID- 6287707 TI - [Safety of foot-and-mouth disease vaccines in cell cultures]. AB - Experiments were carried out to test imported and home-produced production and experimental series of foot- and-mouth vaccines in cell cultures. It was found that the primary cell cultures of swine kidney were most appropriate to study the innocuity of the F. M. D. vaccines, which, in terms of sensitivity proved of superior quality as compared to the primary cell cultures of calf kidney and the BHK 21 cells. Comparative investigations have revealed that the most promising method for testing the innocuity of the F. M. D. vaccines is the elution of the antigen from the aluminium hydroxide after Dannacher and coll. The problem is discussed of using cell cultures to check the innocuity of F. M. D. vaccines prior to their release. PMID- 6287708 TI - [Effect of amantadine (1-aminoadamantan) on different phases of the replicative cycle of transmissible gastroenteritis virus of swine]. PMID- 6287709 TI - [Cultivation of various avian viruses in pheasant trachea organ cultures and chick embryos]. AB - Stationary tracheal organ cultures of pheasants and chick embryos, treated with a mucolytic agent were used to study the ciliostatic effect of the following viruses: fowl pest (FPV), Newcastle disease (NDV), infections laryngotracheitis (ILV), and infections bronchitis (IBV) - strain Beaudette. In chick embryo tracheal cultures ciliostasis was found to set in as follows: for FPV - at the 24th hour; for NDV - at the 72nd-120th hour; for ILV - at the 168-192nd hour; and for IBV-at the 72nd-96th hour. The ciliostatic effect produced by NDV and IBV coincided in terms of time and dynamics. In pheasant tracheal organ cultures ciliostasis set in as follows: for FPV - at the 24th hour; for NDV - at the 72nd 120th hour; and for ILV - at the 168-192nd hour. This system was shown to be unsusceptible to IBV which produced no effect whatever. In mixed organic cultures, containing several tracheal segments of pheasants and chick embryos each per petri dish IBV did bring about ciliostasis in the chick embryo segments only, within the period after inoculation as cited above. It is stated that pheasant tracheal organ cultures could be employed in the case of an express initial differentiation of FPV, NDV, and ILV, recording the time period for the setting in of full ciliostasis following the inoculation of the respective viruses. Discussed is the possibility of identifying IBV, using mixed tracheal organ cultures of pheasants and chick embryos. PMID- 6287710 TI - [Effect of different types of diet on various biochemical aspects of digestive processes in sheep]. AB - Two groups of female lambs of one and the same age, breed, and live weight were fed rations differing in type from two weeks to one year of age. The first group were fed concentrates predominantly, and the second group were fed feeds in which roughage prevailed. After the completion of one year of age experiments and biochemical investigations were carried out with both groups. The intestinal content was studied with regard to amino acid nitrogen and glucose, and the enzyme content - with regard to amylase, lipase, protease, and cellulase in the following sections of the digestive tract: reticulum, rumen, abomasum, small intestine, and caecum. An experiment was also carried out with the resorption in isolated intestinal folds of the initial portions of the jejunum and the caecum in which artificial chyme was introduced. Studied was the resorption of water, chlorides, calcium, glucose, fatty acids, and amino acid nitrogen. Differences were found in the enzyme content and composition of the intestinal content of the various portions of the digestive tract as well as in the resorption with the two groups under the effect of feeding of rations of different type. PMID- 6287711 TI - [Attempts at isolating a precipitating antigen from a cell culture of bovine virus diarrhea-mucosal disease virus]. PMID- 6287712 TI - Hepatocellular carcinoma in an urbanised black community. A changing pattern. AB - Hepatocellular carcinoma has been extensively studied in Southern Africa, and particularly its relationship to hepatitis B virus infection. Most of this work involved rural Black populations. In this study the impact of urbanization on this relationship is investigated. The material is derived from the laboratory records at Baragwanath Hospital which serves the Black urban community of Soweto, Cases autopsied during the periods 1956-1960 and 1976-1980 have been examined with regard to age, sex, underlying cirrhosis and presence of HBsAg in the non tumour liver. In addition, all biopsies from 1955 to 1980 have been analysed with respect to age and sex. There is evidence of a significant increase in overall average age and in the proportion of female cases, while the percentage of HBsAg positive cases has fallen. Possible causes for these findings are considered. PMID- 6287713 TI - Adenoid cystic carcinoma of minor salivary glands. Analysis of 86 cases. Clinico pathological, histoenzymological and ultrastructural studies. PMID- 6287715 TI - Kinetics of incorporation of Sendai virus proteins into host plasma membranes and virions. PMID- 6287714 TI - Giant cell tumor bone. Enzyme histochemical, biochemical and tissue culture studies. AB - Three giant cell tumors of bone (2 benign and 1 malignant) were examined enzyme histochemically, and a tissue culture study of the malignant case was performed. Multinucleated giant cells and mononuclear round cells had similar activities of ACPase and non-specific esterase with a diffuse strong reaction. ATPase and 5' nucleotidase reactions were strongly positive in the cytoplasm of multinucleated giant cells, and were seen not only in the cytoplasm but also on the cell membrane of round cells. The proliferating spindle cells in the malignant case were faintly positive for ACPase and non-specific esterase and were less positive for ATPase and 5'-nucleotidase on the cell membrane. The multinucleated giant cells and mononuclear round cells resembled histiocytes in the activities of 4 hydrolytic enzymes, and the multinucleated giant cells had enzyme activities similar to those of osteoclasts from new-born rat skull. The malignant giant cell tumor and cells in its tissue culture showed ALPase activity preferentially on the cell membrane of the spindle cells, and rarely on round cells or multinucleated giant cells. ALPase was resistant to heat treatment and was found to be the type IV isoenzyme by diffusion electrophoresis. The origin of the giant cell tumor of bone and the significance of the ALPase activity are discussed. PMID- 6287716 TI - Molecular-biological characterization of Marek's disease virus. I. Identification of virus-specific polypeptides in infected cells. PMID- 6287717 TI - Molecular-biological characterization of Marek's disease virus. II. Differentiation of various MDV and HVT strains. PMID- 6287718 TI - Epstein-Barr virus induction by a serum factor. I. Induction and cooperation with additional inducers. PMID- 6287719 TI - Restriction of vesicular stomatitis virus in a nonpermissive rabbit cell line is at the level of protein synthesis. PMID- 6287720 TI - Nucleotide sequence conservation at the 3' termini of the virion RNA species of New World and Old World arenaviruses. PMID- 6287722 TI - Morphogenesis of sandfly viruses (Bunyaviridae family). PMID- 6287724 TI - The cells horizontal cells talk to. PMID- 6287723 TI - Temperature-sensitive B mutants of SV40 disassemble intracellular encapsidation particles at elevated temperature. PMID- 6287721 TI - Characterization of avian erythroblastosis virus p75. PMID- 6287725 TI - [Circulating immunocomplexes in lung neoplasms]. PMID- 6287726 TI - [Current possibilities of scintigraphic detection of non-ischemic myocardial diseases]. PMID- 6287728 TI - [Acute gastroenteritis epidemic in adults caused by rotaviruses]. PMID- 6287727 TI - [A comparison of the pyrophosphate and glucoheptonate heart scan in 3 women with secondary cardiomyopathy (a preliminary report)]. PMID- 6287729 TI - [Use of electrovacuum therapy in diseases of the peripheral nervous system]. PMID- 6287730 TI - [Significance of biochemical indices of liver fibrosis]. AB - Correlation between content of hydroxyproline in urine, protein-bound hydroxyproline, alpha 2-macroglobulin as well as of collagen peptidase activity in blood serum and the intensity of collagen accumulation in liver tissue were studied in experimental liver tissue cirrhosis, which developed after chronic intoxication of rats with CCl4. Distinct relationship was shown between the level of hydroxyprolinuria and amount of collagen in liver tissue. Content of protein bound hydroxyproline, alpha 2-macroglobulin and the collagen peptidase activity were quite unaltered in blood serum during the cirrhosis development. Protein bound hydroxyproline and alpha 2-macroglobulin were markedly decreased in blood serum at the step of decompensated cirrhosis. Among the biochemical patterns studied in biological fluids only estimation of hydroxyproline in urine might distinctly reflect the intensity of liver tissue fibrosis. PMID- 6287732 TI - [Current problems of ambulatory care for cancer patients]. PMID- 6287731 TI - [Superoxide radical and superoxide dismutase in the free-radical theory of aging (a review)]. AB - Review is presented on the role of superoxide radicals and other intermediates of single electron reduction of oxygen in aging as well as on geroprotective importance of superoxide dismutase. These factors are considered in relation with age. Significance of direct and indirect methods for estimation of total O2 radicals is analyzed. Possible role of O2-radicals and superoxide dismutase in regulation of cell division and differentiation is also discussed. Life span of cells in peripheric circulation correlates with superoxide dismutase content in these cells and their ability to generate exogenous O2-. PMID- 6287733 TI - [Lactate dehydrogenase isoenzyme content of the tissue of a cancerous tumor and polyps of the large intestine]. AB - Electrophoretic assays of LDH-isoenzymes in agar were carried out in 599 homogenates of bioptic and resected tissue material obtained from carcinoma (174), polyps (305) and grossly normal mucosa (120). A significant considerable increase in cathode fractions of LDh4 and LDH5 matched by a decrease in LDH1, LDH2 and LDH3 levels was observed in both tumor and polyp tissue. The degree of the changes varied greatly depending on patients' age, type of tumor and size of polyp. Preoperative telegamma therapy was followed by a significant decrease in LDH5 and an increase in LDH2 levels. PMID- 6287734 TI - [Cyclic nucleotides in clinical oncology]. PMID- 6287735 TI - [Information in support of barring the use of tetracyclines and streptomycin as growth stimulants]. PMID- 6287736 TI - [Nutrition and biliary calculi]. PMID- 6287738 TI - [State of the peripheral blood in the combined treatment of lung cancer]. PMID- 6287737 TI - The significance of blood transfusion in non-A, non-B chronic liver disease in Japan. AB - We performed a follow-up study on 70 patients with acute non-A, non-B (NANB) posttransfusion hepatitis and a retrospective study on 283 chronic hepatitis, 70 cirrhosis and 53 hepatocellular carcinoma patients of type NANB. In acute NANB post-transfusion hepatitis, as judged by the transaminase levels, th duration of the disease exceeded 6 months in 46/70 = 65.7% and 1 year in 32/70 = 45.7%. The histological diagnosis of the 32 cases persisting for more than 1 year was chronic active hepatitis in 5, chronic persistent hepatitis in 2 and unresolved hepatitis in 6. The frequency of previous transfusion in chronic hepatitis, cirrhosis and hepatocellular carcinoma of type NANB was 42.8, 37.1 and 15.1%, respectively, whereas the incidence of early posttransfusion hepatitis was 8.5, 8.6 and 7.5%, respectively. in chronic liver diseases with a history of jaundice and/or hepatitis, previous transfusions are more frequently associated with type NANB than with type B disease. The present study demonstrates that NANB posttransfusion hepatitis tends to develop to chronic liver disease when analyzed prospectively as well as retrospectively. PMID- 6287740 TI - [Suloctidil (sulocton) in the treatment of atherosclerotic neurasthenic syndromes]. PMID- 6287739 TI - Hip fracture, skeletal fragility, osteoporosis and hormonal deprivation in elderly women. AB - Fractures of the hip have been shown to have a significant personal and societal impact in Western countries; this impact is largely borne by elderly women, and represents a substantial health care commitment in modern society. For many people a fracture of the proximal end of the femur represents a preterminal event of considerable cost, both in economic loss and psychosocial well-being. These fractures are generally recognized as a clinical complication of osteoporosis, and are one index of general skeletal fragility which is also manifested in fractures of the vertebrate and of the distal radius (Colles fracture). There is increasing evidence that hormonal deprivation in elderly women is directly related to loss of skeletal integrity and consequent fragility. There is also increasing evidence that hormonal substitution is effective in preventing this structural loss and fragility. Unfortunately, a therapeutic dilemma has arisen in that the preparation that seems to give optimal protection, conjugated estrogens, has also been reported to cause an increased incidence of endometrial carcinoma. The search for a preparation or dosage regimen of estrogens which simultaneously prevents skeletal atrophy and fragility and avoids the increased risk of malignancy must be a long-term goal. PMID- 6287741 TI - Tetrahydrocannabinol (THC) for cancer patients. A review of the National Cancer Institute program in Wisconsin. PMID- 6287742 TI - Nutritional value of plantix (fiber) in animal feeding. PMID- 6287743 TI - Plantix (fiber) deficiency in animal physiopathology. PMID- 6287744 TI - Epstein-Barr virus in cerebrospinal fluid during infectious mononucleosis encephalitis. AB - Epstein-Barr virus (EBV) was recovered from cerebrospinal cells (but not from cell-free fluid) of a patient with meningoencephalitis complicating infectious mononucleosis. This finding suggests that virus-altered B lymphocytes can invade the central nervous system and may play a direct role in the pathogenesis of neurologic sequelae of EBV infections. Several mechanisms by which these cells can cause neurologic manifestations are discussed. PMID- 6287746 TI - [Therapy of breast neoplasms. Short overview of some recent developments]. PMID- 6287745 TI - Regulation of bistability in glucose metabolism of Escherichia coli ML 30 chemostat cultures by cyclic AMP. AB - Evidence is presented that cyclic AMP is engaged in the regulation of a bistability in the glucose and energy metabolism of NH3-limited chemostat cultures of Escherichia coli ML 30. Cyclic AMP probably reverses the repression of the citric acid cycle by glucose favouring the state of glycogen and energy overproduction. PMID- 6287747 TI - [Evaluation of various methods of determining crystalline silicon dioxide in comparison with results of industrial dust analysis]. PMID- 6287748 TI - [Cytomegalovirus infections following kidney transplantation - clinical and serological studies of the early and late phases]. AB - In 71 patients after kidney transplantation the cytomegalovirus-antibody state was recognized with the help of the indirect fluorescence antibody test in a period of 24 months. The first estimations were performed in 33 patients in the early phase up to the 3rd month and in 38 patients in the late phase up to the 100th months after transplantation. Of 13 patients who had been controlled already before operation only 3 patients were seronegative. After this twice a seroconversion with clinically manifest cytomegalovirus infection appeared, in one case an irreversible failure of the graft developed. In the late phase 4 patients remained seronegative. Of these patients also in one case the chronic rejection caused the entering into the dialysis programme. -- A positive cytomegalovirus-antibody state was found in the early phase in 30 of 33 patients and in the late phase in 34 of 38 patients. An active cytomegalovirus infection was present in the early phase in 11 of 30 and in the late phase in 11 of 34 patients. In the early phase the clinical symptoms fever, leukopenia and hepatitis were more frequent and more expressed than in the late phase. In 7 of the 11 patients in the early phase and in 8 of 11 patients with active cytomegalovirus infection in the late phase rejections occurred which in 2 of the 7 patients in the early phase and in 5 of the 8 patients in the late phase led to the loss of the graft. In inactive cytomegalovirus infection an irreversible course thrice appeared in 11 patients with rejections. Three typical instances are demonstrated: 1. The course of an active cytomegalovirus infection in the early phase with rejection and irreversible failure of the graft. 2. The reactivation of a latent cytomegalovirus infection by uncontrollable rejection processes. 3. The course of an active cytomegalovirus infection without clinical complications and with transition into an inactive stage in minimal immunosuppression. The treatment is performed with immunosuppression of a possibly low dosage, the avoidance of increases of prednisolone in cytomegalovirus-associated rejections, the intravenous application of human-gamma globulin as well as the prevention or intensive treatment of superinfections. In these cases the close relations between rejection processes, immunosuppressive therapy, superinfections and cytomegalovirus infections should find the necessary consideration. PMID- 6287749 TI - [Atypical variant of a hepatocellular carcinoma with a favourable course (author's transl)]. AB - The case of a 51 years old man with a hepatocellular carcinoma (HCC) is reported. Though the tumor is of multifocal origin and widespread the disease takes a favourable course. Histology revealed a well differentiated HCC with clusters of polygonal tumor cells and fibrous stroma, the pattern of which reminds of focal nodular hyperplasia (FNH). HCC has developed in a non cirrhotic liver. Clinically, the patient is in a good condition 22 months after diagnosis of the tumor. The value of all possible methods--invasive and not invasive--for diagnosis of HCC is assessed and compared with literature. According to the literature the survival time comes to 3 to 6 months on average, only single cases take a more favourable course. Correlation of morphology and biological behaviour of tumor is lacking. In our case the pattern of fibrosis similar to FNH is striking and points to a development of carcinoma in preexisting FNH. In literature, however, evidence for a malignant transformation of FNH is lacking. PMID- 6287751 TI - Measurements with ionselective electrodes in the brain cortex during a short period of ischemia and arterial hypoxia. PMID- 6287750 TI - [Alpha 1-antitrypsin deficiency and the liver (author's transl)]. AB - Alpha 1-antitrypsin (alpha 1 AT) is a glycoprotein of hepatic origin which functions as a systemic protease inhibitor (Pi). Its production is controlled by two autosomal-codominantly transmitted alleles. Among the numerous genetic variants some alleles (predominantly PiZ) may induce alpha 1 AT-deficiency, facultatively associated with childhood liver disease. However, the pathogenesis of this congenital disorder, which may progress to complete cirrhosis remains obscure at present. In addition, no clear cut relationship has been proven between alpha 1 AT-deficiency and deranged liver architecture, observed in advanced aged adults. Possibly this may reflect a more accidental coincidence with the consequences of chronic viral hepatitis (Non A-Non B-type). Nevertheless, this hypothesis is hitherto unestablished as it holds for the supposed association between alpha 1 AT-deficiency and the occurrence of malignant hepatoma. PMID- 6287752 TI - Functional morphology of thyrotropic and corticotropic cells during reproductive cycle in ewe. AB - A histological examination has been carried on the pars distalis of the pituitaries of twelve, healthy sexually mature balady ewes which were classified into three groups Resting, follicular and luteal phases according to the different phases of reproduction. The histological features of the TSH- ACTH cells during resting phase has been used to establish a standard with which the subsequent changes during the follicular and luteal phases were compared. Using PAS/OG, AB/OG and PFA/AB/PAS/OG stains the TSH- and ACTH-cells were differentiated and identified from the other forms of the pars distalis cells. The topographical study of the TSH- and ACTH-cells in various regions and levels in the pars distalis of balady ewe as well as their frequencies during the different phase of reproductive cycle were discussed. Statistical analysis of the different cell count during the different phases of reproductive cycle was conducted. TSH- and ACTH-cells exhibited signs of increased cellular activities and significantly increased in number during follicular phase and in the other phases of the reproductive cycle. PMID- 6287753 TI - Anomalous reduction of cytochrome b in highly purified complex III from baker's yeast. AB - In highly purified bc1-complex from baker's yeast, the reduction of cyt c1 and partial reduction of cyt b is obtained by catalytic amount of succinate dehydrogenase and succinate in the presence of 7 microM antimycin. After the addition of ferricyanide the c1 is re-oxidized and a increase in the reduction of b is observed. Using stopped-flow we established that the oxidation of c1 by ferricyanide proceeds as a pseudo-first order reaction and the reduction of b is faster and with two phases. Our observation suggests that these two processes are not directly interconnected and that other component than c1 must be the "control factor" in the anomalous reduction of cyt b. This component must be, by exclusion, the iron-sulfur protein. PMID- 6287754 TI - [Hormone receptor-adenylate cyclase membrane complex and its functional formation in ontogeny]. PMID- 6287756 TI - [Effect of cyclic adenosine monophosphate on the elaboration of active avoidance in rats]. PMID- 6287755 TI - [Receptive field plasticity of a polyfunctional neuron in mollusks]. AB - Plastic properties of the receptive field of the identified LPa3 neurone were studied on a semi-intact snail preparation. Mantle area representing part of the vast receptive field of the LPa3 neurone of the left parietal ganglion consists of three zones. The right part of the mantle is innervated by the right pallial and anal nerves; the former transmits to the CNS the short latency rapidly extinguishing component of the synaptic response, while the anal nerve--the slowly extinguishing long latency component. The left part of the mantle is innervated by the LPa3 neurone through the left pallial nerve which transmits the multi-component slowly extinguishing excitatory synaptic potential. The pneumostome area is innervated by the right pallial and anal nerves which transmit a non-extinguishing response. Thus, different areas of the mantle are linked to the LPa3 neurone by a system of parallel channels differing by their plastic properties. PMID- 6287758 TI - [Incidence and distribution of benign soft tissue tumors]. PMID- 6287757 TI - [The proportion of N-acetyl-neuraminic acid in human parotid and submandibular gland saliva (author's transl)]. PMID- 6287759 TI - Hepatitis A and B markers and presumable non-A, non-B hepatitis in a psychiatric institution. AB - The frequencies of hepatitis A and hepatitis B markers were determined in 291 members of the staff and 714 patients of a psychiatric institution for adults. 71.8% of the patients suffered from psychosis, 17.7% from neurosis, and 10.6% from oligophrenia. 84.4% of the patients and 69.2% of the staff were anti-HAV positive. - 26.8% of the patients and 19.9% of the staff showed at least one HBV marker. The frequency of anti-HBc increased with age but not with hospitalization. Anti-HBc-positive persons did not show transaminase abnormalities more often as reported previously. - 2.6% of the patients were presumable Non-A, Non-B hepatitis cases after excluding IgM-anti-HAV, IgM-anti CMV or -EBV and possible toxic effects. The rate of presumable Non-A, Non-B hepatitis equaled the frequency of HBsAg-positive persons. PMID- 6287760 TI - Presumable non-A, non-B hepatitis in hemodialysis. AB - Hepatitis A and B markers, IgM-anti-CMV and -EBV were determined in 154 dialysis patients, 118 relatives of patients and 42 members of the staff. Episodes of elevated transaminases were investigated for presumable non-A, non-B hepatitis as well as secondary infections among relatives. --The cumulative frequency of HBV markers was 71% in center dialysis patients, 63% in home dialysis patients, 50% in staff and 19% in relatives. The frequency of HBV markers increased with duration of dialysis treatment and with the number of blood units transfused. Spouses of dialysis patients had more often HBV markers than other relatives (22% vs. 8.8%). 96.8% of the center dialysis patients, 83.3% of home-dialysis patients, 41% of the staff, and 42% of the relatives were anti-HAV-positive. The incidence of IgM-anti-CMV was highest in home-dialysis patients (15.5%). The frequency of IgM-anti-EBV was 4.3% in center-dialysis patients and 12.7% in home dialysis patients. --9% of all dialysis patients had an episode of elevated transaminases compatible with non-A, non-B hepatitis. The possible secondary attack rate among relatives was 31%. PMID- 6287762 TI - Pseudo vitamin D deficiency rickets in pigs: in vitro measurements of renal 25 hydroxycholecalciferol-1-hydroxylase activity. PMID- 6287761 TI - [Devitalization and haemostasis by thermal destruction--results of enzyme histochemical and histological examination of oviduct specimens, following tissue coagulation by means of endocoagulation or high-frequency current (author's transl)]. AB - Semm's technique of endocoagulation has been used by the authors for haemostasis in endoscopic abdominal surgery at the Department of Gynecology and Obstetrics, University of Kiel since 1973. --Enzyme-histochemical and histological tests, all based on endocoagulation or high-frequency current procedures, were applied to 100 human fallopian tubes. Fifteen of them remained uncoagulated and were used for comparison. --Membrane ATPase, 5-nucleotidase, cytochrome-C-oxydase, and other essential enzymes in cellular metabolism were inactivated by temperatures in excess of 57 degrees C. These enzymes, consequently, were no longer detectable from enzyme-histochemical preparations, whereas active tissue regions, those which still contained vital cells, were stained black to brown. --Negative enzyme reactions occurred in response to a coagulation forceps temperature of 120 degrees C, when applied not less than 20 seconds. Residual enzyme activities were still recordable from certain tubal areas, when forceps temperatures below 120 degrees C had been used. In such cases coagulation was insufficient, particularly in the inner layers of tubes (epithelium, mucosa). Temperatures above 130 degrees C and coagulation lengths of more than 20 seconds proved to be unnecessary, since no improvement in results was thus achievable. Temperatures of 140 degrees C and more have changed coagulated tissue into "adhesive" and make it stick to the coagulation forceps. Instrument withdrawal can in such cases cause bleeding rather than the desired haemostatic effect. PMID- 6287763 TI - Adenovirus infection in lambs. III. Studies on chronic infection. PMID- 6287764 TI - [Status, potential and expediency of legal protection regarding plasmids and vectors]. PMID- 6287765 TI - [So-called oxidase test, its mechanism and terminology]. PMID- 6287766 TI - [Nature of the IR spectra of intact cells of Gram-negative microorganisms]. AB - The character of changes in the infrared spectra of E. coli in the process of their mechanical disintegration has been studied. The destruction of E. coli cell structures has been shown to produce no changes in the optical density of the main analytical absorption bands in infrared spectra. This fact suggests that the infrared absorption spectra of E. coli are the sum of the spectra of all chemical components of the cell, which is confirmed by the infrared spectral study of E. coli cell-wall preparations. Similar results have been obtained in the study of the preparations of B. pertussis cell walls, protoplasts and intact cells. PMID- 6287767 TI - [Transparent solid nutrient medium for studying the lytic spectrum of bacteriophages of microbes of the genus Bordetella]. AB - A solid, transparent culture medium for the study of the lytic spectrum of the phages, active against B. pertussis and B. bronchiseptica, in respect to homologous and heterologous bacteria of the genus Bordetella has been developed. The Cohen-Wheeler liquid medium with nicotinic acid and nicotinamide added, solidified with agar, is nicotinamide added, solidified with agar, is used as the base of the new medium. This base ensures the growth of B. parapertussis and B. bronchiseptica. To stimulate the growth of B. pertussis, the tissue stimulant of B. pertussis growth (a transparent substrate obtained from the tissue of the large intestine of a rabbit) has been used. With 10% of this stimulant added, B. pertussis cells have been found to preserve their typical morphological and immunobiological properties. PMID- 6287768 TI - [Isolation and purification of restriction endonuclease BpeI from Bordetella pertussis]. AB - New restriction endonuclease has been isolated from Bordetella pertussis vaccine strain 305 and purified in 1 stage on Sepharose covalently bound with blue dextran. The isolated restrictase has been found capable of breaking down lambda phag DNA into 7 fragments. According to its specificity, Bpe I is the isoschizomer of Hind III obtained from Haemophilus influenzae strain Rd. PMID- 6287769 TI - [Isolation of Vibrio eltor mutants producing melanin and the mapping of the mel locus using transposons]. AB - Melanin-producing V. eltor mutants obtained by means of the transposon which determines resistance to tetracycline (Tn 10) are described. Gene mel is believed to be localized on the chromosome of V. eltor in the region of markers his trp met ura rif arg ilv. PMID- 6287772 TI - The activities of elastase and other lysosomal enzymes in professional phagocytes and mechanism of their release. AB - Differences in specific activities of elastase, myeloperoxidase, and lysozyme were found between polymorphonuclear leukocytes (PMNs) and macrophages of various species, between PMNs derived from different anatomic sites and between macrophages isolated in different ways. The percentage of extracellular release of lysosomal enzymes during immunological phagocytosis depends on the receptors in the phagocyte membrane which were stimulated by binding of a ligand. These differences may reflect various functional activities of separate populations of phagocytes. PMID- 6287771 TI - Receptor mediated endocytosis of hemoglobin-haptoglobin, galactosylated serum albumin and polymeric IgA by the liver. AB - Labelled hemoglobin-haptoglobin (ham-hap), galactosylated serum albumin (gal-SA) and polymeric immunoglobulin A (p IgA) were injected intravenously to rats or mice. The labels disappeared from the plasma with a half-time of about 5 min and were almost entirely found associated with the liver where degradation products progressively appear. The uptake of hem-hap and gal-SA are partially saturable as a function of the plasmatic concentration and the uptake of gal-SA can be completely inhibited by the simultaneous injection of asialofetuin. About 45 min after injection to rats, labelled material appears in the bile in amounts corresponding to 3.9% of the injected dose (hem-hap), 2.8% (gal-SA) and 60.1% (p IgA). The molecular weight of the labelled material transferred into the bile has been characterized: it consists almost entirely of intact IgA and for about 60% of intact hem-hap and gal-SA. Cell fractionation experiments indicate that 4 min after injection, the label is associated with components which equilibrate around a density of 1.13 g/cm3 and which dissociate from marker enzymes of Golgi complex, plasma membrane and lysosomes. Longer times after injection (from 20 min for hem-hap and gal-SA to 1 h for p IgA) labelled material appears, within lysosomes. To explain all these data, we suggest that after binding to plasma membrane receptors, the ligands are rapidly interiorized into pinocytic vesicles which fuse with lysosomes. Most of the hem-hap and gal-SA molecules but only part of p IgA would be released and subsequently digested; these vesicles would dissociate from lysosomes and fuse with the biliary membrane where the molecules still bound to the membrane of the vesicles would be detached and excreted into the bile. PMID- 6287770 TI - [Antigenic similarity of the pertussis component of adsorbed DTP vaccine to human erythrocytes]. AB - The similarity of the heterogeneous antigens, types A and B, of human red blood cells to the most of B. pertussis strains constituting the pertussis component of commercial batches of adsorbed DPT vaccine has been established. This property makes the vaccine strains different from B. pertussis isolated from pertussis patients. One of the reasons of the insufficient effectiveness of immunization against pertussis has been determined: the intensity of immune response depends on the antigenic heterogeneity of the pertussis component of the vaccine and the AB0 group factors in the blood of the vaccinees. For the first time the accumulation of immune alpha- and beta-isoagglutinins in the blood of persons immunized with absorbed DPT vaccine has been established. This accumulation shows the medium degree of direct correlation with the manifestations of the clinical reaction to the injection of the vaccine. The data obtained in this study indicate the necessity to revise the existing method of obtaining the pertussis component of adsorbed DPT vaccine on solid culture media with human red blood cells added and to develop the technique of the additional purification of this component from heterogeneous antigens. PMID- 6287773 TI - Plasma membrane glycoproteins of tumour cells in enzootic bovine leukosis compared with normal bovine lymphoid cells. AB - Plasma membranes from tumor cells in enzootic bovine leukosis and from normal bovine lymphoid cells of different sources have been isolated and characterized. The glycoprotein composition of the different membranes has been studied by SDS PAGE and by analysis of the carbohydrate composition as well as by lectin binding of glycoprotein fractions obtained by phenol treatment of the membranes. No differences in the electrophoretical glycoprotein pattern could be detected comparing (nonmalignant) cells from persistent lymphocytosis and tumour cells, suggesting that malignancy is not associated with the gain or loss of a major glycoprotein. A 151.5 kD glycoprotein, present in membranes from normal lymph node lymphocytes, seems to be absent in the membranes of peripheral blood lymphocytes as well as of tumour cells. The loss of this glycoprotein might thus be associated with a loss of sessility of bovine lymphoid cells. The carbohydrate analysis and lectin binding of extracted glycoproteins revealed a decreased fucosylation accompanied by an increased exposure of galactose residues as well as a loss or a decreased complexity of N-glycosidically bound oligosaccharides in the tumour cell glycoproteins compared with those of normal cells. These findings are discussed with regard to disturbed growth regulation of leukosis tumour cells. PMID- 6287774 TI - Placental secretion of prolactin, ACTH and immunoreactive beta-endorphin during pregnancy. AB - The placental secretion of Prl, ACTH and immunoreactive (IR)-beta-endorphin were examined in early and later pregnancy with a perfusion method and by determining tissue concentrations of these hormones. Prl was secreted mainly from decidual tissue during 5-6 h long perfusions. Tissue concentrations of Prl were also significantly higher in decidual than in trophoblastic tissue. In contrast, the tissue concentrations of hCG, hPL, ACTH and IR-beta-endorphin were significantly higher in decidual than in trophoblastic tissues. In contrast, the tissue concentrations of hCG, hPL, ACTH and IR-beta-endorphin were significantly higher in trophoblastic than in decidual tissues. Production of hCG and hPL was predominantly demonstrated in trophoblastic tissues by the perfusion method but ACTH and IR-beta-endorphin were too low to be detected by perfusion. PMID- 6287775 TI - Concentrations of 2-hydroxyoestrogens in human sera measured by a heterologous immunoassay with an 125I-labelled ligand. AB - A heterologous immunoassay for 2-hydroxyoestrogens has been established in which antibodies raised against 2-hydroxyoestradiol-17-succinyl-BSA serve as binding protein and 2-hydroxyoestrone-17-cmo-[125I]iodohistamine as radioligand. Lipophilic serum components competing for binding sites in this system were defined as total 2-hydroxyoestrogens'. The underlying assumption of specificity was supported by the pattern of cross-reactivity evaluated with structural related steroids and o-diphenols and by the fact, that an additional chromatography of the serum extracts preceding the competing reaction had little if any effect. Sensitivity: 2.8 +/- 1 pg/tube; accuracy: Y = 0.91x + 2.2; r = 0.989; precision: 5.8% intra-assay; 6.5% inter-assay. The following concentrations (+/- standard deviation) were found in the sera of healthy subjects. Young men: 29 +/- 5 pg/ml (n = 11); women follicular phase: 32 +/- 8 pg/ml (n = 25); luteal phase: 53 +/- 13 pg/ml (n = 23); postmenopausal women: 13 +/- 4 pg/ml (n = 10); pregnant women 11th--20th week: 70 +/- 16 mg/ml (n = 64); 36th--40th week: 240 +/- 23 pg/ml (n = 40); newborn cord blood: 604 +/- 43 pg/ml (n = 48). PMID- 6287776 TI - Experimental induction of C cell tumours in thyroid by increased dietary content of vitamin D3. PMID- 6287777 TI - 17 alpha-hydroxylase deficiency syndrome associated with bilateral streak gonads and impaired development of Mullerian ducts derivatives. Report of a case. AB - We report severe 17 alpha-hydroxylase deficiency in a 17 year-old black girl with 46,XX genotype. The diagnosis was suspected because of primary amenorrhoea, absence of sexual maturation, hypertension and hypokalaemia with renal potassium wasting. Endocrine investigation revealed low basal levels of all steroid hormones which require 17 alpha-hydroxylation for biosynthesis (i.e. glucocorticoids, androgens and oestrogens). No increase in their basal levels was seen following adrenal stimulation, indicating a severe block. Plasma concentrations of ACTH, FSH and LH were elevated as were progesterone, 11 deoxycorticosterone and corticosterone. Plasma renin activity was suppressed and aldosterone levels were very low. After 4 months of glucocorticoid replacement therapy, aldosterone was still low, even though the suppression was otherwise effective. Our case is unusual because bilateral streak gonads and impaired development of Mullerian ducts derivatives were also present. To our knowledge, a similar case has never been reported before. PMID- 6287778 TI - Familial elevations of total and free thyroxine in healthy, euthyroid subjects without detectable binding protein abnormalities. AB - Elevations of serum thyroxine without thyrotoxicosis or binding protein abnormalities have been documented in 8 of 13 family members, representing 4 generations. This syndrome appears to represent an elevated threshold for the amount of free thyroxine substrate required to maintain adequate T3 production form the peripheral monodeiodination of T4. It reiterates the need for a prudent re-evaluation of all clinically euthyroid patients with elevated serum thyroxine concentrations before concluding that they are indeed thyrotoxic. PMID- 6287779 TI - Defects in intrathyroid binding of iodine and the perchlorate discharge test. AB - The kinetics of [123I]iodide uptake were studied when organification of iodine by the thyroid gland was normal and when this binding function was diminished by drugs or disease. Each study was terminated by a sodium perchlorate discharge test (300--600 mg iv) at 60 min or, in some cases, 10--30 min. The results confirmed that binding takes place rapidly in the uninhibited gland with the binding rate constant being at least 0.150 min-1. Discharge from the uninhibited gland is less than 3.5% of the gland uptake when perchlorate is given 60 min after the radioiodide. Subjects with an intrinsic binding defect manifested discharges of 11% of greater of the 60 min uptake and the estimated binding rate constants ranged from 0.003--0.057 min-1. Thyrotoxic subjects receiving 5 mg carbimazole twice daily manifested discharges ranging from 5.4--64.2%, and in those receiving 20 mg twice daily the observed discharges were 67.6--94.6% of the 60 min uptake. The study shows that a correctly performed perchlorate discharge test will detect minimal inhibition of iodine binding. An important factor is the duration of the follow-up period after perchlorate is given. In some of the cases studied discharge was not complete until 60 min after the perchlorate. PMID- 6287780 TI - Standardization of the radioreceptor assay against anti-thyrotrophin receptor antibodies in Graves' disease. PMID- 6287781 TI - 2-chloroadenosine increases calcium mobilization from mouse calvaria in vitro. AB - The effect of 2-chloroadenosine on bone resorption and on cyclic AMP formation in murine calvarial bones in vitro was investigated. 2-Chloroadenosine increased the release of 45Ca from the cultured bones, but had no effect on dead bones, indicating that the effect is cell mediated. The adenosine analogue remained in the medium for 48 h and caused a transient stimulation of the formation of cyclic AMP. The dose-response curve for the stimulatory effect on cyclic AMP accumulation was linear up to 10(-4) M. The dose-response curve for 45Ca release was liner from 3 x 10(-7) M to 3 x 10(-5) M but then showed a decline in the response. 8-Bromo cyclic AMP inhibited the release of 45Ca in 24 h cultures. The initial stimulatory effect on bone resorption by 2-chloroadenosine may therefore not depend on cyclic AMP. The level of inosine increased during culture indicating that adenosine is formed by bone tissue. PMID- 6287782 TI - Growth hormone, prolactin and cortisol nyctohemeral variations during naloxone induced opiate receptor blockade in man. AB - To evaluate the role of endogenous opioid peptides in prolactin (Prl), growth hormone (GH) and cortisol neuroregulation, 50 mg of the opiate antagonist naloxone was infused over 24 h to 6 normal male volunteers. An additional naloxone dose (5 mg) was given iv as a bolus injection at 20.00 h. blood specimens were collected hourly by means of a portable constant withdrawal pump. Naloxone failed to alter 24 h secretion of GH and Prl. The sleep-related GH and Prl rise was also unaffected by the opiate blocker. Moreover, naloxone failed to alter the circadian rhythm of cortisol and its 24 h concentration. The results do not suggest, a major role of opiate receptors in spontaneous GH, Prl and cortisol secretion in man. PMID- 6287783 TI - Parathyroid hormone bioassay using human kidney cortical cells in primary culture. AB - The biological activity of parathyroid hormone (PTH) has been investigated by measuring intracellular accumulation of cyclic AMP (cAMP) in human kidney cortical cultures. Enzyme dispersed cortical cells from non-invaded kidney poles of patients undergoing nephrectomy for cancer were used after 5 days of primary culture. Bovine PTH (1-84) produced a significant increase of cAMP accumulation in cultured cells at a dose (53.7 ng/ml) 10-fold lower than that found for the minimal stimulatory effect when using preparations of human purified plasma membranes. The action of bovine PTH (1-84) was very rapid, a response was detected after 5 min and a ceiling effect after 30 min. Cortical cells showed a slightly lower sensitivity to synthetic bovine PTH (1-34) (half maximal increase dose: 0.66 microgram/ml), compared to bovine PTH (1-84) (half maximal increase dose: 0.32 microgram/ml), but revealed a higher sensitivity to human PTH purified from the medium of parathyroid cell cultures (half maximal increase dose: 11.2 ng/ml). Arginine-vasopressin (AVP) also increased the cAMP accumulation of kidney cortical cultured cells, with a potency and efficacy lower than that of human 'culture' PTH, while in kidney medullary cells in primary culture AVP exerted a strong response and the effect of PTH was poor or absent. Calcitonin and glucagon were weak stimulators of kidney cortical cell cAMP accumulation. PMID- 6287784 TI - The effects of pituitary hormones on hepatic drug metabolism in the rat. AB - Sex differences exist in the hepatic microsomal metabolism of drugs in the rat. These differences have been shown to be related to the presence of androgen in the male and the pituitary gland in the female. The present study was designed to identify the pituitary 'feminizing factor' by continuously infusing hormones via an osmotic mini-pump. Rat somatotropin and prolactin mimicked somewhat the 'feminizing' effect of pituitary extract infused into male animals while human somatotropin, which binds to both lactogenic and somatogenic receptors in rat liver, completely reversed the effect of hypophysectomy of female rats with respect to most of the parameters studied. It thus appears that the somatogenic/lactogenic receptors in the rat liver are involved in the maintenance of the sex differences in drug metabolism but the identity of the hormone that normally acts on these receptors in the rat is still uncertain. PMID- 6287785 TI - Cortisol, 17 alpha-OH, progesterone and androgen responses to a standardized ACTH stimulation in different stages of the normal menstrual cycle. AB - The release of cortisol, 17 alpha-OH-progesterone, androstenedione and testosterone during a standardized ACTH-stimulation test was investigated in three different stage of the normal menstrual cycle, to conclude if there is any stage dependency on the release of these hormones. No statistically significant differences were observed between the three stages concerning cortisol and testosterone increase. The increase of androstenedione in the pre-ovulatory stage was significantly high than that seen during the early follicular phase of the cycle. The increase of 17 alpha-OH-progesterone in the luteal phase was significantly less than that of both the early and late follicular stages of the cycle. Progesterone levels showed a small, but significant increase after ACTH stimulation, in both the early and late stage of the follicular phase. However, the levels remained within the normal range of the follicular phase. In the luteal phase no increase was seen after ACTH-stimulation. Oestradiol-17 beta levels did not change at all after ACTH stimulation. The stage dependency of androstenedione and 17 alpha-OH-progesterone is discussed. The described stage dependency different increase of 17 alpha-OH-progesterone release can be of importance when the results of ACTH-tests are evaluated to detect carriers of congenital adrenal hyperplasia. PMID- 6287786 TI - Inhibition of rat follicular androgen synthesis by dibutyryl cyclic AMP. PMID- 6287787 TI - Electron microscopic observation of the red pulp of the spleen in hereditary hemolytic anemia with erythrocyte pyrimidine 5'-nucleotidase deficiency: evidence for intravascular hemolysis. PMID- 6287788 TI - Antianalgesic action of thiamylal sodium in cats. AB - The effects of thiamylal on the nociceptor-driven neural activity in the spinal cord were studied in decerebrate, non-anesthetized cats. Noxious stimulation was induced by the injection of bradykinin into the femoral artery and the neutral response in the lateral funiculus was measured by the multi-unit activity technique. The effects of thiamylal on the bradykinin-induced response were compared before and after the spinal cord transection, above the recording site. Thiamylal, 5 mg/kg i.v., potentiated the response significantly before the cord transection and depressed it after the transection. These findings indicate that the antianalgesic action of thiamylal is induced at the spinal cord level: although this anesthetic agent does have a direct intraspinal depressant action, the multisynaptic neural network of the supraspinal pain inhibition system is more susceptible to the actions of anesthetics, and the depression of this descending system by thiamylal results in a release of spinal cord nociceptive neural mechanisms from the supraspinal control. PMID- 6287789 TI - Naloxone does not antagonize the anesthetic-induced depression of nociceptor driven spinal cord response in spinal cats. AB - The effects of several anaesthetics on spinal cord nociceptive neural mechanisms and their interactions with the opiate antagonist, naloxone, were studied in acute, spinal cord transected cats. Intra-arterial injection of bradykinin was used as the noxious test stimulus. Spontaneous activity and the neural response induced by bradykinin were recorded by the multi-unit activity technique in the lateral funiculus of the spinal cord. Naloxone, 0.1 or 2.0 mg/kg i.v. had little effect on the bradykinin-induced response, but enhanced the spontaneous firing of the lateral funiculus significantly. Fentanyl, 30 micrograms/kg i.v., depressed both the bradykinin-induced response and spontaneous firing. These effects of fentanyl were antagonized completely by naloxone, 0.1 mg/kg i.v. Nitrous oxide, thiamylal, halothane and ether depressed the bradykinin-induced response considerably, but it was not antagonized by naloxone, 0.1-2.0 mg/kg i.v. Enflurane had little effect on the bradykinin-induced response. The effects of these anesthetics on spontaneous firing were divergent: nitrous oxide enhanced it while other drugs depressed it, to various degrees. All these data suggest that the neural and/or neurochemical mechanisms of anesthetic-induced analgesia differ from mechanisms related to opioids. PMID- 6287790 TI - Comparison of intra-and extracellular buffering of clinically used buffer substances: tris and bicarbonate. AB - A large and important group of acid-base disturbances are the metabolic acidoses. In general, every type of metabolic acidosis can be treated with infusion of base when the underlying cause of the disturbances is removed. In our medical centers, the use of tris and bicarbonate is common. For a long time they were competitive agents and until now it was not possible to decide by available clinical methods which of these substances was more suitable for correction of metabolic acidosis. The intracellular pH of the whole rat (mean lcf-pH) was determined from the distribution of 14C labelled DMO (5,5-dimethyl-2,4-oxazolidinedione) and monitored for 6 h following intravenous application of tris or sodiumbiarbonate in a dose of 10 mmol per kg body mass. Arterial plasma pH and PCO2 were also measured. To determine and compare the effectiveness of the two buffer substances, intra- and extracellular bicarbonate were calculated from the Henderson-Hasselbalch equation. It was found that the buffering following bicarbonate infusion is more effective in both body compartments. Sodiumbicarbonate should be preferred in daily practice. PMID- 6287791 TI - Normalization of muscle transmembrane ionic fluxes induced by taurine in patients affected with myotonic dystrophy. PMID- 6287792 TI - The ACTH-secreting system in myotonic dystrophy. PMID- 6287793 TI - Electron spin resonance studies of erythrocyte ghost cells in Huntington's chorea. AB - There is accumulated evidence which suggests that the primary gene defect in the autosomal dominant disease of Huntington's Chorea, is given expression as a generalised membrane abnormality in peripheral tissues. Several publications claim to be able to detect a difference between HC patients and controls by means of the electron spin resonance (ESR) technique. We have examined the electron spin resonance spectra of the spin probe 4-maleimido-2,2,6,6 tetramethylpiperidinooxyl (MAL-6) when incorporated into the membranes of erythrocyte ghost cells of 22 patients with HC and 47 controls, in 2 series of controlled, blind studies and we were unable to detect any difference between HC patients and normals. We conclude that the ESR technique with the probe used is not a sufficiently reliable test for accurate differentiation of HC patients from controls and is certainly not suitable as a method for the diagnosis of potential carriers of this disease. PMID- 6287794 TI - Allergic reactions to steroids presenting with neurological symptoms. PMID- 6287795 TI - Clinical and virological features of herpes genitalis in Japanese women. AB - We report herein clinical and virological findings in 35 Japanese women with a confirmed herpes genitalis. The frequency of herpes genitalis was 0.24% in our obstetrical and gynecological outpatient clinic, while that in cancer screening clinics was 0.01%. Eighty percent of all patients were between 20 and 49 years of age (median age of 38.5 years). Genital herpes was detected more frequently in summer and autumn. Virus isolations were carried out in 20 patients and 13 were positive (65.0%). Of 13 genital isolates with positive cultures, 7 (53.8%) were identified as herpes simplex virus type 1 (HSV-1) and 6 (46.2%) as herpes simplex virus type 2 (HSV-2). Two virgins were diagnosed virologically and cytologically as having progenital herpes, and in these 2 women, HSV-1 was isolated by the virus isolations. Lack of antibodies to either virus was higher in HSV-1 type infection. These studies together with previous findings show that both clinical and virological features of genital herpes differ between Japanese and Caucasian or black women. These differences are attributed to some extent to sexual patterns of behavior in the different races. PMID- 6287796 TI - A tumor composed of myofibroblasts: an ultrastructural study. AB - A subcutaneous tumor of the right temporal region of head was removed from an 8 year-old boy. This tumor was studied by light and electron microscopy. He was found to have had a retroperitoneal inflammatory fibrous histiocytoma which had been excised seven years ago. The subcutaneous tumor histologically consisted of spindle cells in fascicles, which did not arrange in a storiform or herring-bone pattern. Electron microscopically, this tumor was chiefly composed od myofibroblasts. electron microscopy is essential to determine the cell types of a tumor and we have classified this tumor as a tumor of myofibroblasts. The relationship between the retroperitoneal tumor and the head tumor was discussed. PMID- 6287797 TI - Fibrous histiocytoma of the nasal cavity and maxillary sinus. AB - Two cases of fibrous histiocytoma were presented; Case 1: a 46-year-old man with a tumor filling the left nasal cavity and maxillary sinus, and Case 2: an 80-year old man with a tumor of the right maxillary sinus destroying its surrounding wall. Histologically, the former was dominated by storiform pattern and onion like structures, and the latter by osteoclast-like multinucleated giant cells. After radical surgery, Case 1 was free of recurrence for more than one year, and Case 2 showed recurrence although still maintaining fairly good health. A total of 29 cases hitherto reported arising in the nasal, paranasal and nasopharyngeal regions were reviewed, pitfall of differential diagnoses was commented especially in our two cases, criteria of malignancy were discussed, and the necessity of repeated samplings was emphasized because of paucity and unfamiliarity of fibrous histiocytoma arising, in particular, from the upper respiratory tract. PMID- 6287798 TI - Adenoma with multiple hyperplastic nodules in noncirrhotic liver possibly related to long-term administration of a hypolipidemic drug. AB - Described here is an autopsy case of a 76-year-old woman with preneoplastic and other adenomatous hepatocellular lesions. Multiple small hyperplastic liver foci with PAS positive reaction and adenomatous nodular lesions were noted in a non cirrhotic liver. These lesions resembled altered foci or neoplastic nodules which are currently regarded as premalignant changes in experimental animal models. It is suggested that the liver lesions of this woman may be related to long-term administration of hypolipidemic drug clofibrate for hypercholesteremia and the lesions could be one of the precursor changes of human hepatocellular carcinoma. PMID- 6287799 TI - Bronchiolo-alveolar adenocarcinoma with multiple cysts. AB - An autopsy case of bronchiolo-alveolar adenocarcinoma in the lung is reported. The patient is a 70-year-old male who complained of severe cough with 500-600 ml watery sputum a day, loss of weight. and general fatigue. Autopsy revealed numerous whitish tumors in various sizes with multiple cysts in both lungs, with no metastasis being found in any other organs. Histological findings identified the tumor as a bronchiolo-alveolar adenocarcinoma originating from the lungs. Electron-microscopic findings showed that the tumor cells were covered by prominent microvilli, and contained abundant irregularly-shaped cytoplasmic vacuoles suggestive of mucin. PMID- 6287801 TI - Production of reactive oxygen species and chemiluminescence by human monocytes during differentiation and lymphokine activation in vitro. AB - The zymosan-induced respiratory burst of human monocytes was studied, measuring the generation of reactive oxygen species (ROS) and luminol-dependent chemiluminescence (CL). The monocytes' ability to generate ROS declined markedly during the first 24 hours of culture, but regained the original level during the following three days. In contrast, the CL-response declined steadily during the same period. In vitro activation by lymphokines from lymphocytes stimulated with bacillus Calmette-Guerin gave a significant increase in ROS-generation; the CL response was only marginally increased, but activated cells consistently displayed an altered kinetics of the CL-response with a more rapid decline from peak CL-values. These findings indicate that the production of ROS is not the limiting factor for the magnitude of the CL-response in the present experimental system and that the CL-assay may be an uncertain way of quantitating ROS. PMID- 6287800 TI - Properties of Fc gamma receptors in the human central nervous system. AB - Cryostat sections of human central nervous system (CNS) tissue absorbed sheep erythrocytes (E) sensitized with rabbit IgG antibody (A). The indicator cells bound to the choroid plexus, to the leptomeninges covering the brain and spinal cord, to the arachnoid granulations and to the perivascular tissue of the neural parenchyma. Unsensitized E or E sensitized with F(ab')2 fragments of IgG were not bound. The reactions were inhibited by pooled human IgG, human IgG1 and IgG3 myeloma proteins, and Fc fragments of pooled human IgG. Whole human IgG2 myeloma protein, IgM, IgA, F(ab')2 fragments of pooled human IgG and albumin did not inhibit. Experiments with reduced and alkylated IgG, EDTA, iodoacetamide, 2 mercaptoethanol, various pHs and salt concentrations, formaldehyde, periodic acid and neuraminidase did not reveal any differences between the Fc gamma receptors in the separate anatomical areas. The Fc gamma receptors in human CNS are thus apparently very similar. PMID- 6287802 TI - [Changes of action potential in human papillary muscles by methoxyverapamil (author's transl)]. PMID- 6287803 TI - [Effects of dl-demethylcoclaurine on electrophysiological properties of porcine myocardial cells (author's transl)]. PMID- 6287804 TI - [Analysis of blocking action of dimethyl dl-curine dimethochloride on neuromuscular transmission (author's transl)]. PMID- 6287805 TI - Vascular smooth muscle relaxation by nitro compounds: reduced relaxation and cGMP elevation in tolerant vessels and reversal of tolerance by dithiothreitol. AB - Smooth muscle preparations obtained from bovine mesenteric artery were incubated with high concentrations of nitroglycerin (NG) or nitroprusside (NP) at elevated pH. This treatment was found to make the vessels tolerant towards both the relaxant and cGMP elevating action of a challenging dose of nitrocompounds when tested on the histamine contracted vessels. Also, some cross-tolerance between NP and NG could be found since pretreatment of the vessels with NP caused a reduction in both the relaxant and the cGMP elevating action of NG. The NG induced tolerance could be partly reversed by treatment with the disulfide reducing agent dithiothreitol (DTT). These results are suggested to strengthen the evidence for cGMP as a mediator of vascular smooth muscle relaxation induced by nitrocompounds. The ability of DTT to partly reverse this tolerance indicates the existence of critical tissue sulfhydryl groups with which the nitrocompounds interact. There also seemed to exist certain differences in the mechanism of action between NG and NP since the tolerance induced against NG was much more pronounced than was the case for NP. PMID- 6287806 TI - Dose dependent induction of rat liver microsomal cytochrome P-450 and microsomal enzymatic activities after inhalation of toluene and dichloromethane. AB - Sprague-Dawley rats were exposed, by inhalation, to toluene and dichloromethane (500, 1,500 or 3,000 p.p.m.) and to benzene (1,500 p.p.m.) for three days. Toluene and benzene increased the concentration of liver microsomal cytochrome P 450. A dose dependent increase in the in vitro liver microsomal formation of several metabolites of biphenyl and benzo(a)pyrene was observed for both dichloromethane and toluene. At the highest dose-level the increase in the vitro formation of benzo(a)pyrene-7,8-dihydrodiol was more than three-fold for both dichloromethane and toluene whereas the formation of benzo(a)pyrene-4,5 dihydrodiol increased more than five-fold following exposure to toluene but less than two-fold after exposure to dichloromethane. Our results suggest that dichloromethane and toluene can modify the metabolism and thereby the toxicity of other environmental contaminants. PMID- 6287807 TI - Effect of polyunsaturated fatty acids of the alpha-linolenic series in the lipid composition of rat testicles during development. AB - The effect of 22:6 omega 3 acid provided by dietary fish oil on the lipids composition of rat testes and liver during the development of the germinal tissue, was investigated. Results were compared to those obtained in animals fed on sunflower seed oil (linoleate) and methyl palmitate. At 7 and 9 weeks of age, both fish oil and methyl palmitate evoked a decrease of phosphatidyl-choline and ethanolamine and an increase of triacylglycerols. However, only in the group of rats administered with omega 6 or omega 3 acids and not in the palmitate group, the maturation of germinal epithelium was normal. It is suggested, as in a previous work, that 22:6 omega 3 may functionally replace 22:5 omega 6 in rat germinal tissue. PMID- 6287808 TI - [Isolation of a membranous fraction of toad gastric mucosa]. AB - In the present work the isolation and some characteristics of a membrane fraction from toad gastric mucosa are described. By combining differential centrifugation in a discontinuous gradient of sucrose it was possible to obtain a band at the interphase of 22% sucrose 37%, whose enzymatic and electron microscopic analysis indicated to be a fraction enriched of luminal plasma membranes from oxyntic cells, with very poor contamination by other subcellular organelles. This is a suitable membrane preparation that can be used to perform studied on the mechanisms of acid secretion in toads. PMID- 6287810 TI - Intraerythrocytic sodium and potassium concentrations during acute and chronic digitalization. AB - To assess the cellular effects of digoxin, intraerythrocytic sodium and potassium concentrations were measured in 17 patients during the early phase of digitalization, in 45 patients on long-term therapy and in 64 non-digitalized control patients. Acute digitalization raised intraerythrocytic sodium from 11.6 +/- 0.4 to 16.7 +/- 1.0 mmol/l (mean +/- SEM) (p less than 0.01) and reduced intraerythrocytic potassium from 100.1 +/- 1.3 to 95.9 +/- 1.8 mmol/l (p less than 0.01). These changes were strongly correlated with the steady-state plasma digoxin concentration. During a few weeks of digoxin therapy, the intraerythrocytic cation composition normalized gradually. In patients on chronic treatment, neither intraerythrocytic sodium (11.3 +/- 0.3 mmol/l) nor potassium concentrations (100.0 +/- 0.6 mmol/l) differed significantly from the values of the control group (11.4 +/- 0.2 and 99.9 +/- 0.5 mmol/l, respectively). The changes in intraerythrocytic cation concentrations, induced by acute digitalization, seem to disappear during chronic administration of the drug. PMID- 6287809 TI - Diagnosis of endocrine gastrointestinal tumours. AB - In 32 patients with endocrine gastrointestinal tumours angiography, CT, ultrasound examination, and in some cases phlebography with blood sampling for hormone analyses were compared with respect to diagnosis of the primary tumour and possibly existing liver metastases. Primary tumours in the pancreas and liver metastases could be diagnosed with CT and ultrasound, except for two small insulinomas, which required angiography. PMID- 6287811 TI - Urinary cyclic AMP corrected for glomerular filtration rate in the differential diagnosis of hypercalcemia. AB - To evaluate the usefulness of urinary cyclic AMP (U-cAMP) expressed as nmol/100 ml glomerulus filtrate (GF) when discriminating various hypercalcemic states, we studied 99 patients. Patients with primary hyperparathyroidism (PHPT) showed a positive correlation between individual S-calcium levels and U-cAMP, nmol/100 ml GF (females r=0.49, n=40, p less than 0.01 and males r=0.91, n=7 p less than 0.001). There was also a correlation between U-cAMP, nmol/100 ml GF, and the weight of the adenomas (females r=0.36, n=32, p less than 0.05) and males r=0.79, n=6, p less than 0.05). Patients with PHPT and normal renal function excreted more U-cAMP than controls, 6.0 +/- 1.6 versus 4.3 +/- 1.0 nmol/100 ml GF (mean +/ SD). Of 47 patients with PHPT and normal renal function, 29 showed values below the upper normal limit, 6.3 nmol/100 ml GF (mean +/-2 SD), of the control group; the overlap was 62%. When U-cAMP was expressed as mumol/24 hours, the overlap was 40/47 (85%) and, when expressed as mumol/g creatinine, 31/47 (66%). Three patients with sarcoidosis and two with malignancies and hypercalcemia showed excretory values of U-cAMP, nmol/100 ml GF, above the upper normal limit. Patients with acromegaly or prolactinoma showed normal values of U-cAMP, nmol/100 ml GF. The present data indicate that all three types of determinations of urinary cAMP based on 24 hour urine collections are of little value in the differential diagnosis of hypercalcemic states. PMID- 6287812 TI - On the enzymatic make-ups of the integumentary derivatives and their physiological roles in the Indian centipede, Scolopendra moristans. PMID- 6287813 TI - Columnar organization, local circuitry and synaptology of the mammalian cerebral cortex. PMID- 6287814 TI - Peripheral neuropathy in plasmacell dyscrasia with IgMk paraprotein production. PMID- 6287815 TI - Effect of cyclic AMP-dependent protein kinases on gene expression. PMID- 6287816 TI - Phosphorylation of cAMP-dependent protein kinase subunits. AB - The cAMP-dependent protein kinases comprise two enzyme forms designated as type I and type II. The type II enzyme can catalyze an autophosphorylation reaction whereby phosphate is transferred from ATP to one seryl residue on each regulatory subunit monomer. Since this reaction can occur in the absence of cAMP-induced enzyme dissociation, it has been used as a probe to identify one site of interaction between the catalytic subunit (C) and the type II regulatory subunit (R11). The type I cAMP-dependent protein kinase does not catalyze an analogous reaction; however, if cGMP-dependent protein kinase is substituted for C, the type I regulatory subunit (R1) becomes phosphorylated. The effects of cyclic nucleotides on this reaction, coupled with the ability of R1 to serve as an inhibitor of cGMP-dependent protein kinase suggest that this phosphorylation also occurs within an important functional domain on R1. A comparison of the autophosphorylation site on R11 with the cGMP-dependent protein kinase catalyzed phosphorylation site on R1 indicates that each modification takes place within a similar proteolytically sensitive region. On each subunit, this sensitive "hinge" region lies distal to the functional domain responsible for regulatory subunit dimerization and proximal to that responsible for cAMP binding. Phosphorylation of the "hinge" region decreases the affinity of each regulatory subunit for C, although the magnitude of this change appears greater for R1 than for R11. Phosphorylation of R1 also reduces the stoichiometry of cAMP binding from two to one mole of cAMP bound per mole of R1 monomer. These results suggest that the "hinge" regions of both R1 and R11 form part of the interaction site between the regulatory subunit and C; and, in the case of R1, it also forms a portion of one of two cAMP-binding sites. The amino acid sequence surrounding the phosphorylated serine of each regulatory subunit has been determined: R11: D-R-R-V-S(P)-V R1: R R-R-R-G-A-I-S(P)-A It is thought that the number and position of the basic amino acid residues proximal to the modified serine may be responsible, in part, for determining the susceptibility of each site to phosphorylation by cAMP or cGMP dependent protein kinase. Both R1 and R11 exist as phosphoproteins in vivo. Dephosphorylation of purified "native" phospho-R1 is without effect on the ability of R1 to interact with either C or cAMP. The site phosphorylated in vivo is therefore distinct from that modified in vitro by cGMP-dependent protein kinase. In addition to the autophosphorylation site, R11 possesses a second, less enzymatically reactive, phosphorylation site that is modified in vivo. Dephosphorylation of this site is also without apparent effect on the functional properties of R11. The kinases responsible for catalyzing the phosphorylation of R1 and the cryptic site on R11 and the role that these modifications play in modulating kinase activity are currently unknown but are under active investigation. PMID- 6287818 TI - Biological chelation: 2,3-dimercapto-propanesulfonic acid and meso dimercaptosuccinic acid. AB - Water soluble analogs of British Anti-Lewisite that are active orally and less toxic than BAL are now available. These agents are 2,3-dimercapto-1 propanesulfonic acid and meso-dimercaptosuccinic acid. Evidence for their effectiveness in preventing the lethal effects of sodium arsenite in mice and lewisite in rabbits is presented. These analogs can be expected to replace BAL in the treatment of heavy metal poisoning. PMID- 6287817 TI - Regulation of cytochrome P450 activities in adrenocortical mitochondria from normal rats and human neoplastic tissues. AB - From our immunocytochemical studies, cytochrome P450scc and P45011 beta systems were localized on the matrix side of inner membrane of the mitochondria in the parenchymal cells of adrenal cortex. However, the degree of immunocytochemical staining varied from one mitochondrion to another within a single cell; some stained intensely along the entire inner membrane, some stained only along segments of the inner membrane, and some did not stain at all. The ratio of stained to unstained mitochondria was approximately unity in untreated rats, while stained mitochondria greatly increased upon ACTH administration suggesting that the population of cytochrome P450 system-containing mitochondria increased upon long-term ACTH action. By the combined use of flash photolysis and substrate difference spectroscopy, quantitative determination of substrate-bound and free forms of P450scc and P45011 beta in mitochondria became possible. Increases in total amounts of P450scc and cholesterol resulted from the long-term ACTH administration to rats. The ratio of cholesterol-bound P450scc to its free form was essentially unchanged under these conditions. On the other hand, amounts of P45011 beta were not increased significantly by long-term ACTH administration. The ratio of DOC-bound P45011 beta to the free form increased significantly, however, by long-term ACTH administration and also in the mitochondria from patients suffering from Cushing's syndrome. PMID- 6287819 TI - Restriction analysis of cloned heteropolymeric DNA synthesized in vitro by TdT. PMID- 6287820 TI - Polyclonal B-cell activators in the study of the regulation of immunoglobulin synthesis in the human system. PMID- 6287821 TI - Intraocular penetration of cefmenoxime (SCE-1365), A new cephalosporin, in humans. PMID- 6287822 TI - Border disease in Norway. Serological examination of affected sheep flocks. PMID- 6287823 TI - Iron studies in patients with sickle cell disease. AB - The prevalence of iron deficiency anaemia during the first three decades of life was investigated in eighty-five patients with SS and SC haemoglobins. The parameters used were the haematocrit, serum iron, total iron binding capacity (TIBC), percentage saturation of transferrin and availability of iron in the bone marrow. The mean haematocrit values were similar throughout the three decades, but increased with age (r = 0.41). The mean serum iron was significantly lower (P less than 0.01) in the first decade than in the second or third decade. Females had lower serum iron in the first and second decades and higher values in the third decade than their male counterparts. The transferrin saturation was significantly lower (P less than 0.01) in the first decade than in the third decade. No haemosiderin was found in the marrow aspirates at a transferrin saturation of less than or equal to 15%. Of the eighty-five bone marrow aspirates studied for stainable iron, fifty-eight (68.2%) had nil iron. The data presented show that iron deficiency anaemia is a common finding in patients with haemoglobinopathies. The need to incorporate oral iron with folic acid and paludrine in the treatment of sickle cell disease is suggested. PMID- 6287824 TI - The value of angiography in the management of stroke. PMID- 6287825 TI - Diseases of peripheral nerves as seen in the Nigerian African. AB - The anatomical and aetiological diagnoses of peripheral nerve disease excluding its primary benign and malignant disorders, as seen in 358 Nigerians are presented. There is a male preponderance and the peak incidence is in the fourth decade. Sensori-motor neuropathy was the commonest presentation (50%). Guillain Barre syndrome was the commonest identifiable cause (15.6%), accounting for half of the cases with motor neuropathy. Peripheral neuropathy due to nutritional deficiency of thiamine and riboflavin was common (10.1%) and presented mainly as sensory and sensori-motor neuropathy. Diabetes mellitus was the major cause of autonomic neuropathy. Isoniazid was the most frequent agent in drug-induced neuropathy. Migraine (20%) was not an uncommon cause of cranial neuropathy although malignancies arising from the reticuloendothelial system or related structures of the head and neck were more frequent (26%). In 26.5% of all the cases, the aetiology of the neuropathy was undetermined. Heredofamilial and connective tissue disorders were rare. Some of the factors related to the clinical presentation and pathogenesis of the neuropathies are briefly discussed. PMID- 6287826 TI - Monitoring foeto-placental function in Nigeria by estimation of urinary oestriol. PMID- 6287827 TI - Maternity care monitoring in Ibadan, Nigeria. AB - During this hospital stay, 993 women delivered 1008 infants at University College Hospital in Ibadan, Nigeria from January through June 1977. The stillbirth rate was 45.4 per 1000 deliveries; the neonatal mortality rate was 20.2 per 1000 deliveries. Significant differences occurred in patient characteristics, antenatal care, and complications of labour/delivery between those patients with favourable and unfavourable birth outcomes. As expected from all previous reports, the rate of multiple births was very high: 37 sets of twins and 3 sets of triplets. Contraceptive acceptance rose after delivery from 17.1% to 63.5% accepting some method. Orals were the most popular choice both antepartum and postpartum. PMID- 6287828 TI - Measles immunity and immunization in developing countries of Africa: a review. AB - Although an effective vaccine against measles has been available for several years, the disease is still prevelant in Africa. The disease is characterized by its occurrence in younger age groups and high morbidity and mortality. The African child is born with a high transplacentally acquired measles antibody level. However, the antibody declines rapidly, so that it is virtually absent after the age of 6 months. The measles vaccine with which the African child is immunized is of reduced potency because of poor storage and transportation facilities and the adverse effect of tropical climate on the vaccine. The pattern of measles immunity in Africa is different from that in developed countries. Measles immunization schedule in Africa, as in any other part of the world, must be based on sound epidemiological and serological data. PMID- 6287829 TI - Controlled comparative study of the efficacy of pyrantel pamoate and a combined regimen of piperazine citrate and bephenium hydroxynaphthoate in the treatment of intestinal nemathelminthiases. AB - Pyrantel pamoate at a dose of 10 mg/kg body weight daily for 2 or 3 days was found to be more effective against Ascaris than both a regimen of bephenium hydroxynaphthoate (2.5 g base) and piperazine citrate (2-3 g) given daily for 2 days and a placebo-treated control group. The cure rates for pyrantel were about 95% compared to 90 and 20% for the other two groups respectively. None of the drug schedules was effective against the Trichuris. The bephenium/piperazine regimen was superior to both the 2-day and 3-day courses of pyrantel pamoate against the hookworm with aggregate cure rates of 58-60% for pyrantel pamoate and 68-85% for bephenium/piperazine. The results were better than those reported earlier in the series when a single dose of pyrantel pamoate (10 mg/kg of body weight) was used. Pyrantel pamoate was well tolerated and the drug merits further studies at higher doses, particularly against the hookworm. PMID- 6287831 TI - Lobar bronchioloalveolar cell carcinoma. AB - Bronchioloalveolar cell carcinoma has a variety of radiographic appearances. Usually it is a localized, well circumscribed nodule in the lung periphery. Nodules of various sizes that appear to coalesce in one or both lungs characterize a diffuse pattern of this disease. Occasionally, chronic lobar consolidation simulating a bacterial pneumonia may be seen. The clinical and radiographic course of eight patients whose bronchioloalveolar cell carcinoma appeared initially as lobar consolidation was analyzed. Four tumors rapidly progressed to involved the opposite lung. Attempts should be made by radiographic and other means to establish inoperability, since surgery is rarely useful in this type of lung cancer. PMID- 6287830 TI - Determination of serum acid and alkaline phosphatase using 4-methylumbelliferyl phosphate. AB - The present report compares the performance of several colorimetric substrates presently employed in clinical laboratories for the determination of serum acid and alkaline phosphatase with that of the fluorogenic phosphatase reagent 4 methylumbelliferyl phosphate. Acid and alkaline phosphatase assays were performed on reference hospital populations in Nigeria and the United States. The results of both acid and alkaline phosphatase determinations indicate that the coefficient of variation is smaller for the fluorometric assay than for the colorimetric methods that employ either phenyl phosphate or p-nitrophenyl phosphate as the substrate. Furthermore, serum acid phosphatase determinations using 4-methylumbelliferyl phosphate identified increased serum enzyme levels in 6/6 cases of prostatic carcinoma and 9/9 cases of Gaucher's disease. The occurrence of elevated serum alkaline phosphatase levels in 24/24 cases involving liver and bone disease was also confirmed by the fluorometric alkaline phosphatase assay. In addition, sera from eight patients with the connective tissue disease, osteogenesis imperfecta, were found to contain normal levels of acid and alkaline phosphatase by the fluorometric assays. From these results it appears that the sensitive and rapid fluorometric assay procedures can be readily employed in the clinical pathology laboratory for the determination of serum acid and alkaline phosphatase levels. PMID- 6287832 TI - Calcium-channel blockers: pharmacologic considerations. PMID- 6287833 TI - Clinical implications of anterior S-T segment depression in patients with acute inferior myocardial infarction. AB - To assess various factors associated with anterior S-T segment depression during acute inferior myocardial infarction, 47 consecutive patients with electrocardiographic evidence of a first transmural inferior infarction were studied prospectively with radionuclide ventriculography an average of 7.3 hours (range 2.9 to 15.3) after the onset of symptoms. Thirty-nine patients (Group I) had anterior S-T depression in the initial electrocardiogram and 8 (Group II) did not have such "reciprocal" changes. There was no difference between the two groups in left ventricular end-diastolic or end-diastolic volume index or left ventricular ejection fraction. Stroke volume index was greater in Group I than in Group II. There were no group differences in left ventricular total or regional wall motion scores. A weak correlation existed between the quantities (mV) or inferior S-T segment elevation and reciprocal S-T depression. No relation between anterior S-T segment depression and the left ventricular end-diastolic volume index could be demonstrated; the extent of left ventricular apical and right ventricular wall motion abnormalities, both frequently associated with inferior infarction, did not correlate with the quantity of anterior S-T depression. These data show that anterior S-T segment depression occurs commonly during the early evolution of transmural inferior infarction, is not generally a marker of functionally significant anterior ischemia and cannot be used to predict left ventricular function in individual patients. Anterior S-T segment depression may be determined by reciprocal mechanisms. PMID- 6287834 TI - Measurement of infarct size using single photon emission computed tomography and technetium-99m pyrophosphate: a description of the method and comparison with patient prognosis. AB - The application of dual tracer transaxial emission computed tomography of the heart was studied with use of technetium-99m pyrophosphate and technetium-99m labeled red blood cells for measuring infarct size in 20 patients with acute myocardial infarction and 10 without infarction. Imaging was performed with a standard gamma camera and with a multidetector transaxial emission computed tomographic body scanner 3 hours after injection of technetium-99m pyrophosphate. Immediately after the scanning procedure, technetium-99m pertechnetate was injected to label red blood cells, and the scanning protocol was repeated. Technetium-99m pyrophosphate was detected in the anterior wall with involvement of the interventricular septum or lateral wall in patients with electrocardiographic criteria for anterior infarction, whereas uptake was detected in the diaphragmatic left ventricular wall with involvement of the posterior, posteroseptal or posterolateral left ventricle or of the right ventricle in patients with electrocardiographic criteria for inferior or posterior infarction. Infarct size measured from transaxial images ranged from 14.0 to 117.0 g in weight. There was a direct relation between infarct size and patient prognosis in that, of the 13 patients with infarct greater than 40 g, 11 (85 percent) had complications, whereas only 2 (29 percent) of 7 patients with an infarct less than 40 g had complications during a follow-up period averaging 17.8 months (p less than 0.05). PMID- 6287835 TI - Vitamin D, hydroxyapatite, and calcium gluconate in treatment of cortical bone thinning in postmenopausal women with primary biliary cirrhosis. AB - Women with primary biliary cirrhosis malabsorb calcium, phosphate and vitamin D, and develop accelerated cortical bone thinning. We have assessed the value of parenteral vitamin D, oral hydroxyapatite (HA), and calcium gluconate (CG) in the treatment of cortical bone thinning in primary biliary cirrhosis. Sixty-four postmenopausal women with primary biliary cirrhosis were assigned randomly into three groups: one group receiving no mineral supplements (control), one group receiving HA, and one group receiving CG. All patients received parenteral vitamin D2 (100,000 IU monthly). Eleven patients withdrew from the study and 10 withdrew due to poor compliance (six HA, four CG). Over a 14-month follow-up period, none of the groups showed a significant change in serum calcium or inorganic phosphate levels. Pre- and posttreatment hand radiographs were used to assess changes in metacarpal cortical thickness using the technique of caliper radiogrammetry. Cortical bone loss occurred in the control group (p less than 0.01). The HA group showed a significant gain in cortical bone thickness (p less than 0.01), while no significant change occurred in the CG group. This study indicated that vitamin D2 does not halt metacarpal cortical bone thinning in primary biliary cirrhosis. The addition of CG prevents bone thinning, and HA promotes positive cortical bone balance. PMID- 6287836 TI - Prognosis in children with Wilms' tumor metastases prior to or following primary treatment: results from the first National Wilms' Tumor Study (NWTS-1). AB - Survival data from the first National Wilms' Tumor Study were analyzed on 101 patients who relapsed after primary therapy (group I-III) and on 82 patients who presented initially with metastases (group IV). The group I patients had significantly better 3-year survival rates than group II/III patients (64% vs 37%). Histology, relapse site, and time to relapse all showed strong independent effects on survival even in the presence of other variables. The survival curves were significantly better for patients with favorable histology, extraabdominal metastases, and late metastases. The overall survival (longer than 3 years) was 46% of group I-III patients who relapsed and 56% of group IV patients. PMID- 6287837 TI - Age as a criterion for eligibility in a lung cancer clinical trial. AB - The results of a clinical trial can be generalized only if the study sample is representative of the population with the disease entity under study. A prerequisite for patient participation in a clinical trial is that all the eligibility criteria be met. In many cooperative group trials, it is common practice to use age as a criterion for eligibility on the premise that advanced age is an unfavorable prognostic factor. In this paper, we evaluate the bias introduced in a sample when this practice was followed in a lung cancer protocol. Of 653 lung cancer cases reviewed, 77 met the eligibility criteria; an additional 23 patients were ineligible for the trial solely on the basis of age. There was no significant difference in survival between these two groups of patients. This indicates that age is not an unfavorable prognostic factor for patients with lung cancer and should not be used as a criterion for eligibility in a clinical trial. PMID- 6287839 TI - Simultaneous evaluation for terminal deoxynucleotidyl transferase and myeloperoxidase in leukemia. AB - A technique for dual staining of cells using terminal deoxynucleotidyl transferase (TdT) and myeloperoxidase (MPO) is described. The technique has been applied to cells of two patients. One patient had chronic myelomonocytic leukemia evolving into acute myelomonocytic leukemia. The other patient had chronic myelogenous leukemia in blast crisis. Our findings indicate that TdT and MPO are exclusive markers except for a rare precursor cell with dual staining in one patient. This study supports the concept of acute mixed leukemia. PMID- 6287838 TI - The effect of bran on bowel function in constipation. AB - Bran is widely used to treat a variety of gastrointestinal disorders. With few exceptions, investigative work has focused on the effect of bran in normal subjects. Despite its widespread use, there have been no systematic studies of bran in constipation. Wheat bran is the "gold standard" because no other natural fiber has yet been shown to be as effective in increasing fecal bulk. We evaluated the effect of two brans, wheat and corn (20 g/day), on fecal weight, fecal moisture content, bowel movement frequency, intestinal transit time, and symptoms in 10 constipated (less than or equal to 3 movements/wk) but otherwise healthy women. The administration of bran was associated with a significant increase in fecal weight (157%), bowel movement frequency (55%), and decrease in intestinal transmit time (50%). Percentage fecal moisture increased only with wheat bran (67.4-72.1%), whereas corn bran was significantly better than wheat bran in relieving symptoms of constipation. PMID- 6287840 TI - Euthyroid familial hyperthyroxinemia due to abnormal thyroid hormone-binding protein. AB - A family is described in which three members had an elevated total serum thyroxine level and free thyroxine index. Each affected subject was clinically euthyroid and had a normal pulse wave arrival time (QKd), serum triiodothyronine and free thyroxine levels, and a normal serum thyroxine-binding globulin (TBG) concentration. Electrophoresis of their serum with 125I-labeled thyroxine revealed increased thyroxine binding in the albumin region. In addition, this abnormal protein, like thyroxine-binding globulin, bound 125I-labeled triiodothyronine and 125I-labeled reverse triiodothyronine. However, electrophoresis of serum treated by sialidase (neuraminidase) digestion suggested that this abnormal protein is not an anomalous form of thyroxine-binding globulin "buried" in the albumin area. These cases of euthyroid familial hyperthyroxinemia due to an abnormal thyroid hormone-binding protein show that an elevated serum thyroxine level or free thyroxine index is not always sufficient to confirm the presence of thyrotoxicosis. PMID- 6287841 TI - Genetic heterogeneity of I-cell disease is demonstrated by complementation of lysosomal enzyme processing mutants. AB - I-cell disease (mucolipidosis II) is a fatal childhood disorder affecting the expression of multiple lysosomal acid hydrolases. The disorder is characterized by clinical and biochemical heterogeneity which may reflect different mutants with a similar phenotype. Genetic complementation studies demonstrating genetic heterogeneity within this disorder are described utilizing cultured fibroblasts from 11 different patients. Fibroblasts from I-cell disease (ICD) and from five different lysosomal storage diseases with single structural gene enzyme deficiencies were fused in different combinations, and fractions enriched for multinucleated heterokaryons were isolated and tested for acid hydrolase activity and electrophoretic mobility. In fusions of ICD fibroblasts and various single lysosomal enzyme-deficient fibroblasts, the activity of the deficient enzyme and of the other ICD hydrolases were restored, demonstrating that ICD is not a lysosomal enzyme structural gene defect and that the ICD defect, and not just the single enzyme deficiency, is corrected. In fusions involving only I-cell fibroblasts, at least two complementation groups were identified by the recovery of activities of all lysosomal enzymes tested in heterokaryons. These results demonstrate the existence of genetic heterogeneity within the disorder and suggest that different mutations can result in the I-cell clinical and biochemical phenotype. The data support an altered post-translational processing of lysosomal enzymes as the cause of ICD and suggest that at least two genes participate in this pathway. PMID- 6287842 TI - Intraepithelial squamous lesions of the vulva: biologic and histologic criteria for the distinction of condylomas from vulvar intraepithelial neoplasia. AB - We reviewed 65 intraepithelial lesions of the vulva and distal vagina and compared the presence of koilocytosis, abnormal mitoses, and parabasal or basal nuclear enlargement with DNA microspectrophotometric distribution patterns and the presence of human papillomavirus antigen as determined by immunoperoxidase. Abnormal mitoses and cytologically atypical nuclear enlargement were specific predictors of aneuploidy and were reliable for distinguishing vulvar intraepithelial neoplasia (VIN) from condylomas. Koilocytosis was present in 100% of condylomas and 71% of aneuploid (VIN) lesions, but there were qualitative and quantitative differences in the distribution of koilocytic cells in the two classes of lesions. On the basis of these findings, criteria for distinguishing between VIN and condyloma are proposed. PMID- 6287843 TI - Treatment of nonmetastatic gestational trophoblastic disease: results of methotrexate alone versus methotrexate--folinic acid. AB - Two treatment regimens for nonmetastatic gestational trophoblastic disease are compared in this retrospective study. The course of 39 patients with nonmetastatic gestational trophoblastic disease treated with methotrexate alone is contrasted to that of 29 patients with nonmetastatic gestational trophoblastic disease who were treated with methotrexate alternated with folinic acid. Of those patients initially treated with methotrexate alone, 7.7% developed methotrexate resistant disease and required a change in chemotherapy for induction of remission. In contrast, 27.5% of patients initially treated with methotrexate and folinic acid developed methotrexate-resistant disease and required a change in chemotherapy to achieve remission. Ultimately, remission was achieved in all patients. Methotrexate as single-agent chemotherapy was found to be consistently more toxic than methotrexate alternated with folinic acid. It is concluded that methotrexate with folinic acid at the dosage used in this study, while less toxic than methotrexate alone, is less effective than methotrexate alone in the induction of remission of nonmetastatic gestational trophoblastic disease. PMID- 6287844 TI - Increased content of Type V Collagen in desmoplasia of human breast carcinoma. AB - Type V (AB) collagen is present in increased amounts in desmoplasia of human breast carcinoma. Type V collagen occurs as linear deposits in the interstitium (desmoplastic stroma surrounding the tumor), as demonstrated by immunofluorescence and immunoperoxidase techniques utilizing antibodies to Type V collagen. In contrast, no demonstratable Type V occurs in the interstitium of fibroadenoma, fibrocystic disease, or normal breast tissue. As identified and quantitated by pepsin extraction, salt fractionation, and polyacrylamide gel electrophoresis, Type V constitutes 10% +/- 5% of all interstitial collagens (Types I, III, and V) in desmoplasia, compared with 1.5% +/- 0.5% in fibroadenoma and less than 0.1% in all types of fibrocystic disease, including sclerosing adenosis, and less than 0.1% in normal breast tissue. For infiltrating ductal carcinoma, Type V collagen is not secreted by the invasive carcinoma cells, nor is it present in an extracellular basement membrane location. By immunohistologic methods, Type V collagen can be seen exclusively in the desmoplastic stroma. The authors propose that desmoplasia of human breast carcinoma manifests a characteristic collagen profile and that the increased Type V may be produced by specialized cells such as myofibroblasts in the interstitium, which are recruited in response to invasive carcinoma. PMID- 6287845 TI - The effect of the beige mutation on infection with murine cytomegalovirus: histopathologic studies. AB - The tissue damage induced by murine cytomegalovirus (MCMV) in mice with the beige mutation (bg/bg) and in their normal littermates (bg/ +) was investigated. The beige mutation in mice is a homolog of the Chediak-Higashi syndrome in man, and various dysfunctions of phagocytes and decreased activity of natural killer cells have been demonstrated in these animals. Tissue damage, especially in the liver and spleen, was more conspicuous in bg/bg than in bg/ + mice and was associated with frequent intranuclear inclusions and a higher titer of virus. However, the mutation did not appear to alter the organ distribution of tissue damage induced by MCMV. The inflammatory response in the liver, which is presumed to contribute to host resistance, appeared under certain circumstances to be delayed and deficient in bg/bg mice. PMID- 6287846 TI - Production and characterization of a monoclonal antibody to human Type IV collagen. AB - We have produced a monoclonal antibody to human basement membrane Type IV collagen. The antibody reacts with the pepsin-resistant, collagenase-sensitive domain of Type IV collagen isolated from placental membranes, but not with human collagens of Types I, II, III, V, 1alpha, 2alpha, and 3alpha. The antibody precipitates biosynthetically labeled human Type IV procollagen, and the precipitate contains both the alpha1 (IV) and alpha2 (IV) chains, suggesting the occurrence of both of these chains within the same triple-helical molecule. When used in indirect immunofluorescence, the antibody gives brilliant staining of basement membranes from a variety of human tissues but does not stain tissues of bovine, canine, rabbit, rat, or mouse origin. It is suggested that this antibody will be of value in research on the structure of human basement membrane collagen, on the distribution of this collagen in various basement membranes, and particularly for the study of basement membranes in normal human development and pathologic processes. PMID- 6287847 TI - Estradiol stimulation of pituitary cAMP production and cAMP binding. AB - The role of 17 beta-estradiol (E2) in the modulation of N6,O2'-dibutyryl adenosine 3',5'-cyclic monophosphate (DBcAMP)-induced hormone release was examined in pituitary monolayer cultures prepared from intact and ovariectomized female rats. Incubations with 5 mM DBcAMP for 4 h significantly (P less than 0.05) stimulated both luteinizing hormone (LH) and prolactin (PRL) release in pituitary cultures prepared from rats at diestrus and from cycling rats at random stages of the estrous cycle. However, DBcAMP failed to stimulate the LH or PRL release in cultures prepared from ovariectomized rats in which the basal LH and PRL release was approximately three-fold and one-tenth of that in cycling rats, respectively. Pretreatment with 1 nM E2 augmented or restored the DBcAMP-induced LH release but not the DBcAMP-induced PRL release in cultures prepared from cycling or ovariectomized rats, respectively. Furthermore, E2 treatment alone of cultures prepared from cycling rats significantly increased intracellular cAMP concentrations and cAMP-binding activities by at least twofold over that of the non-E2-treated controls. The E2-induced rise in cellular cAMP concentration preceded the E2-induced rise in cAMP binding. These results indicate that the priming effect of E2 on pituitary LH responsiveness to DBcAMP is associated with increased cAMP production and cAMP binding. PMID- 6287848 TI - Syndromes of thyroid hormone resistance. AB - Resistance to the action of thyroid hormone can involve both peripheral tissues and the pituitary (global resistance), the pituitary only or peripheral tissues alone. Global resistance is of variable severity and has been observed in more than 60 individuals, the majority occurring in 17 families. Affected subjects are commonly eumetabolic and have goiters, elevated plasma levels of total and free thyroxine and triiodothyronine, normal thyroid hormone metabolism, and normal serum TSH levels (albeit high for the corresponding levels of thyroid hormone). A variable degree of delayed bone maturation, mental retardation, learning disabilities, and hearing defects have been reported; and a variety of treatment regimens, most of which are aimed at reducing the level of plasma hormones and/or goiter, have been attempted before the correct diagnosis has been reached. The clinical disorder is equally common in males and females and appears to be due to one or more autosomal gene mutations. The causes for the hormone resistance may be heterogeneous, either influencing the receptor for thyroid hormones or some unidentified steps in hormone action. At present, the diagnosis is one of exclusion; no effective therapy is available, and all measures designed to lower serum thyroid hormone levels should be avoided. PMID- 6287849 TI - Effect of aging on urinary concentrating mechanism and vasopressin-dependent cAMP in rats. AB - The effect of aging on urinary concentrating ability and the pathogenic mechanism involved were investigated in Fischer 344 rats. While the rats had free access to drinking water, 24-mo-old rats were polydipsic and polyuric compared with 6- and 12-mo-old rats. The maximum urinary concentrating ability after 40-58 h of water deprivation was not different between 6- and 12-mo-old rats (Uosmol 2,941 +/- 173 vs. 2,706 +/- 96 (SE) mosmol/kg), but it was significantly decreased in 24-mo-old rats (1,885 +/- 172 mosmol/kg, P less than 0.01). Similarly, although 5 mU/ml vasopressin increased the concentration of cAMP and papillary slices in 12-mo-old rats (delta +2.81 +/- 0.62 pmol/mg tissue, P less than 0.01), the same concentration of vasopressin failed to increase the cAMP concentration in 24-mo old rats (delta +0.25 +/- 0.21 pmol/mg tissue, P greater than 0.05). In the adenylate cyclase preparation of renal papilla, the response to low concentrations of vasopressin was diminished in 24-mo-old rats. The dose-response curve was shifted to the right and the ED50 concentration of vasopressin was increased in 24-mo-old rats compared with 12-mo-old rats: 1.40 +/- 0.12 mU/ml vasopressin vs. 3.04 +/- 0.22. These results suggest that the decrease in vasopressin-dependent cAMP generation may in part be responsible for the impairment of urinary concentrating ability in 24-mo-old rats. PMID- 6287850 TI - Effects of arginine vasopressin on the thin ascending limb of Henle's loop of hamsters. AB - Arginine vasopressin (AVP) has been shown to stimulate active Cl transport across the medullary thick ascending limb of Henle's loop (MAL) in association with an increase in adenylate cyclase activity. To determine whether the failure to demonstrate active Cl transport across the thin ascending limb of Henle's loop (TAL) in previous in vitro perfusion studies was due to the absence of AVP in the preparation, we examined the effect of AVP on adenylate cyclase activity and Cl transport in the hamsters TAL. AVP (1 mU/ml) increased adenylate cyclase activity in the hamster TAL (20.7 +/- 5.2 control vs. 46.2 +/- 10.1 fmol . mm-1 . 30 min 1, n = 6, P less than 0.05) but not in the descending limb (27.8 +/- 7.0 control vs. 20.4 +/- 2.7, n = 4, P less than 0.05). When both MAL and TAL were perfused, a lumen-positive transepithelial voltage (Vt) was observed. The Vt was increased by adding 1 or 10 mU/ml AVP to the bath. When only the TAL was perfused, the Vt was not different from zero. Similar results were obtained in mouse renal tubules. In other experiments, AVP did not affect the diffusion potential generated when a transepithelial NaCl gradient was present. AVP or dibutyryl cAMP caused little or no change in efflux of radioactive chloride across the hamster TAL. These findings suggest that electrogenic chloride transport is not demonstrable in the TAL even in the presence of AVP. The physiologic role of AVP sensitive adenylate cyclase in the TAL remains to be established. PMID- 6287851 TI - Effect of cycloheximide on corticosteroid-induced changes in colonic function. AB - Chronic parenteral mineralocorticoid and glucocorticoid treatment increases colonic sodium and water absorption and mucosal Na-K-ATPase activity. Cycloheximide, a protein synthesis inhibitor, was utilized to compare the mechanisms of action of these corticosteroids. Rats were injected with 50 or 100 micrograms/100 g body wet cycloheximide every 12 h, 0.5 or 3 mg/100 g deoxycorticosterone (DOCA) daily, or 3 mg/100 g methylprednisolone (MP) daily, singly or in combination for 2 days. In water absorption, transmural potential difference, and the specific activity of Na-K-ATPase were measured. Cycloheximide alone did not alter colonic water, sodium, or chloride absorption or Na-K-ATPase activity but did increase transmural potential difference. DOCA-induced increases in colonic absorption and Na-K-ATPase were completely prevented by cycloheximide. Cycloheximide completely prevented the increase in Na-K-ATPase in MP-treated rats but only partially reduced the MP-induced increase in sodium and water absorption. These results suggest that this enzyme is not the primary site of glucocorticoid action. It remains to be determined whether an increase in Na-K ATPase activity is a necessary part of the maximal colonic response to chronic glucocorticoid treatment. PMID- 6287852 TI - Chloroquine stimulates absorption and inhibits secretion of ileal water and electrolytes. AB - The effects of chloroquine diphosphate, a drug with "'membrane-stabilizing" properties, were studied on basal ileal absorption and on ileal secretion induced by increased intracellular cAMP levels and calcium (serotonin). The studies were performed on rat (in vivo) and rabbit ileum (in vitro). Intraluminal chloroquine (10(-4) M) reversed cholera toxin- and theophylline-induced secretion in rat ileum but did not alter the cholera toxin- and theophylline-induced increases in cAMP content. Addition of chloroquine (10(-4) M) to the mucosal surface of rabbit ileum did not alter basal active electrolyte transport or the serotonin-induced decreased Na and Cl absorption but inhibited the theophylline-induced C1 secretion. Addition of chloroquine (10(-4)) M) to the serosal surface stimulated net Na and Cl absorption. This effect may involve intracellular calcium. Chloroquine increased the rabbit ileal calcium content and decreased 45Ca2+ influx from the serosal surface. Both the mucosal and serosal effects of chloroquine described led to a net increase in absorptive function of the intestine and should prove useful in developing treatment of diarrheal diseases. PMID- 6287854 TI - A mathematical model of primary pacemaking cell in SA node of the heart. AB - Application of voltage-clamp techniques to cardiac primary pacemaker tissue is currently a subject of considerable interest in cardiac electrophysiology. Information regarding the electrical behavior of this type of membrane is by no means complete, yet sufficiently detailed information is available in the literature that would allow the formulation of a reasonably quantitative model to represent the electrical activity of primary pacemaker tissue. In this study the well-known McAllister-Noble-Tsien (MNT) model of the cardiac Purkinje fiber is modified to account for the electrical activity of the primary pacemaking cell (P cell) of the sinoatrial (SA) node in the heart. Modification is accomplished by appropriately deleting (or inactivating) selected inward- or outward-current channels in the MNT model and adjusting the range of voltage dependence of the various channel-gating variables. Wherever possible, this is accomplished by taking into account voltage-clamp and other electrophysiological data from experiments on the SA node. The resultant model mimics published data quite well and is capable of characterizing the free-running behavior of the primary pacemaker as well as its response to injected currents. PMID- 6287853 TI - Characterization of bombesin effects on canine gastric muscle. AB - Synthetic bombesin increased the frequency and amplitude of spontaneously occurring contractions of muscle strips from antrum and corpus of the canine stomach in vitro. The effect of bombesin on frequency of contractions in this system was myogenic; was unrelated to receptors for acetylcholine, norepinephrine, substance P, or cholecystokinin; and was independent of extracellular calcium. The effect of bombesin on the amplitude in the circular muscle was unaffected by tetrodotoxin and atropine but locked by verapamil, whereas in the longitudinal muscle it was blocked by tetrodotoxin and atropine. In the circular antral muscle bombesin was approximately 100,000 times more potent than acetylcholine at threshold concentrations and was equipotent to cholecystokinin. The high potency of bombesin and its known presence in gastric nerve fibers make it a candidate for a neurotransmitter function in regulation of gastric mobility. PMID- 6287855 TI - Relationship of mitochondrial alterations and 99mTc pyrophosphate uptake during myocardial ischemia. PMID- 6287857 TI - Effect of cholera toxin on cAMP levels and Na+ influx in isolated intestinal epithelial cells. AB - Freshly isolated chicken intestinal cells contain approximately 20 pmol adenosine 3',5'-cyclic monophosphate (cAMP)/mg cellular protein. Incubation with 3 micrograms/ml cholera toxin (CT) at 37 degrees C induces an elevation of cellular cAMP beginning 10-15 min after initial exposure. The response is linear with time for 40-50 min and causes a six- to eightfold increase over control levels at steady state. Dibutyryl cAMP and agents that increase cAMP production inhibit Na+ influx into the isolated enterocytes. Chlorpromazine completely abolishes the toxin-induced elevation of cAMP in the isolated cells and also reverses the effect on Na+ entry. The data provide evidence for a cAMP-mediated control of intestinal cell Na+ uptake, which may represent the mechanistic basis for the antiabsorptive effect of CT on Na+ during induction of intestinal secretory activity. Studies on the time-dependent effects of chlorpromazine on both intracellular cAMP concentration and Na+ influx suggest that the reactivation of the Na+ transport system after cAMP-induced inhibition is slow relative to the disappearance of cAMP. PMID- 6287856 TI - Postnatal carbon monoxide exposure: immediate and lasting effects in the rat. AB - Groups of newborn rats inhaled 500 ppm CO for 32 days, after which they continued development in ambient air. The ratio of heart weight to body weight increased sharply after birth, peaked at 14 days of age, and then fell progressively but remained above that of normal rats at 68 and 107 days of age. At 14 days of age, the weight of the left ventricle plus interventricular septum (LV + S) exceeded the controls by 80%, whereas the weight of the right ventricle (RV) was 100% greater. RV weight remained significantly greater than that of the controls at 68, 107, and 217 days of age. The ratio of RV weight to LV + S weight remained higher than that of the controls both during and after CO exposure. Myocardial deoxyribonucleic acid (DNA) concentration (microgram/mg dry wt) declined more rapidly in the CO-exposed groups during the first 2 wk. DNA content (microgram/LV + S + RV) was not significantly different. There were no differences in DNA concentration or content after CO exposure. Hydroxyproline, used as an index of collagen content, was unaffected by postnatal cardiomegaly. Hydroxyproline concentration was depressed only during the first 3 wk. Cardiac cytochrome c concentration was depressed and lactate dehydrogenase M subunit composition elevated only during CO exposure. Neither lactate dehydrogenase specific activity nor myoglobin concentration was altered during or after CO treatment. Neither the DNA nor hydroxyproline data provide an explanation for "persistent cardiomegaly." PMID- 6287858 TI - An ATP- and Ca2+-regulated Na+ channel in isolated intestinal epithelial cells. AB - When isolated intestinal epithelial cells are treated with 2 mM ATP, the unidirectional influx of Na+ to those cells increases from values near 50 to rates over 200 nmol . min-1 . mg protein-1. Calcium influx increases from 1 to 40 nmol . min-1 . mg protein-1. Within 2 min, the total cell Na+ increases two- to threefold, and total Ca+ increases about fivefold. The cells lose a major part of their capability for accumulating sugars during this interval. About 2 min after the time of ATP addition the normal permeability for Na+ and Ca2+ is restored, at which time the previously accumulated ions are rapidly extruded on a net basis until control levels are attained and the cells regain their usual sugar transport capability. The "repair" process requires Ca2+ in the incubation medium and is dependent on cellular uptake of Ca2+. Chlorpromazine (0.5 mM) blocks the Ca2+ entry route and the restoration of normal Na+ permeability. The Na+ entry route is selectively blocked by 4-acetamido-4'-isocyanostilbene-2,2'-disulfonic acid. The data show that ATP induces the influx of Na+ and Ca2+ by two different routes, which can be selectively inhibited. These ion flux routes may be involved in the events that allow intestinal tissue to convert from an absorptive state to a state in which net ion secretion occurs. PMID- 6287859 TI - Increase in K+ and alpha-AIB active transport in CHO cells after low [K+] treatment. AB - We have studied the effects of prolonged incubation in low [K+] medium (approximately 0.3 mM) on both K+ and amino acid transport in Chinese hamster ovary (CHO) cells. When incubated in low [K+] medium, CHO cells redressed partially the loss of intracellular K+ after 12 h. After 24 h of incubation, both the activity of Na+-K+-ATPase in crude homogenates, and the transport capacity (Vmax) for ouabain-sensitive (i.e., active) K+ influx approximately doubled. The magnitude of the ouabain-insensitive (i.e., passive) K+ influx decreased by 50%. Thus the regulatory response involves an apparent increase in Na+-K+ pump and a decrease in K+ leak. The transport capacity for the nonmetabolized amino acid, alpha-aminoisobutyric acid (alpha-AIB), also increased after 24 h in low [K+] medium. The Vmax for the Na+-dependent (i.e., active) alpha-AIB influx increased by about 150%, and the magnitude of the Na+-independent influx increased by 20 40%. These changes in alpha-AIB transport result in a twofold greater capacity to accumulate this amino acid. Thus the regulation of K+ and alpha-AIB transport systems appears to be linked and possible mechanisms of this linkage are discussed. PMID- 6287860 TI - Volume regulation by Necturus gallbladder: apical Na+-H+ and Cl(-)-HCO-3 exchange. AB - Necturus gallbladder epithelial cells exhibited volume regulatory swelling when exposed to a hypertonic mucosal bathing solution. The initial, osmotically induced shrinkage was followed by a rapid increase in cell volume back to the control value despite continuing hypertonicity of the mucosal perfusate. This volume regulatory increase occurred by osmotic water flow accompanying the transient cellular uptake of NaCl from the mucosal bathing solution. Volume regulatory increase required Na+ and Cl- in the mucosal bath; it was inhibited by amiloride or 4-acetamido-4'-isothiocyanostilbene-2,2'-disulfonic acid but not by bumetanide or ouabain. The K1/2 for Na+ was 2.8 mM, the K1/2 for Cl- was 1.9 mM, and maximum velocity of fluid flow into the cell for both ions was greater than 10 x 10(-6) cm/s. Both volume regulatory increase and transepithelial fluid absorption involve NaCl flux across the apical membrane into the cells, but the nature of the NaCl fluxes differ in the two processes. During volume regulatory increase NaCl enters the cells by parallel Na+-H+ and Cl(-)-HCO-3 exchanges, whereas during transepithelial fluid absorption NaCl enters the cell by the coupled flux of NaCl. PMID- 6287861 TI - Stimulation of Na+-Ca2+ exchange in cardiac sarcolemmal vesicles by proteinase pretreatment. AB - The Na+-Ca2+ exchange activity of purified canine cardiac sarcolemmal vesicles can be strikingly stimulated if the vesicles are pretreated with a serine or thiol proteinase. The Km (Ca2+) for Na+i-dependent Ca2+ influx is reduced from 22.2 +/- 2.3 to 8.1 +/- 0.3 microM while Vmax is increased from 15.1 +/- 3.6 to 18.9 +/- 5.2 nmol Ca2+ . mg protein-1 . s-1. Na+o-dependent Ca2+ efflux is also stimulated by proteinase pretreatment although passive (Na+-independent) Ca2+ efflux from the sarcolemmal vesicles is unaffected. Proteinase treatment reduces the sensitivity of Na+-Ca2+ exchange to the inhibitors chlorpromazine and polymyxin B, but not to the inhibitor, palmitylcarnitine. Using a newly developed technique we are able to demonstrate that the Na+-Ca2+ exchange of inside-out sarcolemmal vesicles is being stimulated by proteinase treatment (Philipson, K. D., and A. Y. Nishimoto, J. Biol. Chem: 257, 5111-5117, 1982; this technique uses the ATP-dependent Na+ pump to preload only inside-out vesicles with Na+ prior to Na+-Ca2+ exchange). Right-side-out vesicles may also be stimulated. PMID- 6287862 TI - Abnormal brown adipose composition and beta-adrenoceptor binding in obese Zucker rats. AB - The composition, morphology, beta-adrenergic receptor binding, and in vitro lipolysis were examined in lean and obese, 5- to 6-mo-old male Zucker rat interscapular brown adipose tissue (IBAT). IBAT pads from obese rats were heavier (283%), had more lipid (700%), and more (75%)( and larger (83%) adipocytes than those from lean rats. Also, IBAT from obese rats had no multiloculated cells, and 50% of their IBAT adipocytes were the size of white fat cells. High affinity binding for (-)-[3H]dihydroalprenolol (KD, 15-18 nM), as well as the estimated KD values for binding and the 1/2 Vmax values for adrenergic agonist-induced lipolysis were similar in isolated IBAT cells from lean and obese rats. However, adipocytes from IBAT in obese rats had 75% fewer high affinity beta-adrenergic binding sites per cell (Bmax) compared to those in lean rats. These findings are most compatible with the infiltration of IBAT by white adipocytes. Such infiltration would be expected to reduce the overall thermogenic capacity of IBAT in obese Zucker rats and thereby contribute to the maintenance of their obesity. PMID- 6287863 TI - Increased fetal insulin receptors and changes in membrane fluidity and lipid composition. AB - Fetal tissues exposed to hyperinsulinemia in utero have significantly greater numbers of insulin receptors than do those of controls. We have studied this upregulation phenomenon using crude microsomal membranes from fetal rabbit litters exposed to varying degrees of hyperinsulinemia in diabetic pregnant rabbits. We have observed that insulin binding capacity of membranes increased directly with the severity of maternal diabetes, ranging from 8.5 ng in controls to 44.6 ng insulin/mg membrane protein in offspring of severely diabetic animals and related directly with increasing fetal insulin levels (r = 0.77, P less than 0.005). Lipid analyses of fetal lung membranes showed that reduction of phospholipid to protein ratios occurred in the presence of maternal diabetes. Membrane cholesterol-to-phospholipid ratios were also altered in the presence of maternal diabetes. Significantly, increases in plasma membrane microviscosity were noted in the membranes from diabetic offspring. The data suggest that reduction of membrane fluidity is associated with increases in fetal membrane insulin receptors in severely diabetic pregnancies. PMID- 6287864 TI - Protein wasting due to acidosis of prolonged fasting. AB - During a total fast in obese subjects, the daily rate of nitrogen excretion undergoes only a small further decline after 2 wk, the excretion rate being about 5 g N/day. At this time, ammonium and urea each constitute about one-half of this excretion. The purpose of this study was to consider two alternative hypotheses: first, that the near plateau in nitrogen excretion represents an irreducible minimum rate of net protein breakdown in order to supply essential organs with calories in the form of glucose; second, that protein breakdown could be further reduced by minimizing the requirement to provide nitrogen for ammonium excretion during the ketoacidosis of fasting. Because ammonium excretion is largely controlled by acid-base balance, 150 mmol of sodium bicarbonate plus 60 mmol of potassium chloride were administered daily to decrease ammonium excretion in eight obese subjects who were totally fasting for more than 14 days. Urine ammonium nitrogen fell with this treatment (from 3.8 +/- 0.4 to 2.0 +/- 0.4 g N/g creatinine). In addition, there was a smaller fall in the rate of urea excretion (from 2.5 +/- 0.2 to 2.1 +/- 0.3 g N/g creatinine) together with a fall in the blood urea nitrogen. Therefore, it appears that ammonium excretion contributes to the negative nitrogen balance of a prolonged total fast, as assessed over a 3-day period of observation, is responsible for about one-third of the net lean body mass loss. PMID- 6287865 TI - Structural aspects of adaptive changes in renal electrolyte excretion. PMID- 6287866 TI - Pepsinogen release from isolated gastric glands. AB - The in vitro release of pepsinogen was studied using a preparation of isolated gastric glands from rabbits. The pepsinogen content of the glands was estimated to be about 700 U/mg dry wt. Spontaneous release of pepsinogen was found to be less than 1% of the total per hour and relatively constant for at least 2 h. Pepsinogen release was stimulated in a dose-dependent manner by both carbachol and isoproterenol. The cholinergic and beta-adrenergic stimulation was selectively inhibited by atropine and propranolol, respectively. Removal of external calcium inhibited the responses to both isoproterenol (partially) and carbachol (completely). Several agents, including histamine, prostaglandin (E2), and synthetic secretin, were found not to stimulate pepsinogen release. However, a crude secretin preparation (Boots) was found to produce significant stimulation. Dibutyryl cAMP increased pepsinogen release in a dose-dependent manner. Isoproterenol was found to increase the cAMP content of gastric glands and to stimulate adenylyl cyclase activity in homogenates. The beta-adrenergic stimulation of adenylyl cyclase was found to be selective for a population of gastric cells that was relatively depleted of parietal cells and distinct from the histamine-stimulated adenylyl cyclase activity. The results indicate that pepsinogen secretion by the gastric chief cell is regulated, in part, by separate cholinergic and beta-adrenergic mechanisms and that both calcium and cAMP play a role in this regulation. PMID- 6287867 TI - Acetylcholine inhibits positive inotropic effect of cholera toxin in ventricular muscle. AB - Acetylcholine (ACh, 10(-6) M) had no effect on basal adenylate cyclase activity (3.4 +/- 0.56 pmol cyclic AMP . min-1 . mg wet wt-1), adenosine 3',5'-cyclic monophosphate (cyclic AMP) content (0.88 +/- 0.09 pmol/mg wet wt), or the force of contraction in paced (2.5 Hz) chick embryo right ventricles superfused with Tyrode solution. After 60-180 min of superfusion in the presence of cholera toxin (5 x 10(-6) g/ml), adenylate cyclase activity (1.7 times), cyclic AMP content (2.4 times), and contractility (2.4 times) had increased significantly above basal levels. ACh reversed the positive inotropic effect of cholera toxin but did not change the increased activity of adenylate cyclase and content of cyclic AMP obtained in cholera toxin. Stimulation of adenylate cyclase by isoproterenol (ISO) was inhibited by ACh in the absence and presence of cholera toxin. ACh did not change guanosine 3',5'-cyclic monophosphate (cyclic GMP) content in the absence or presence of cholera toxin. Cholera toxin has actions on chick embryo ventricle similar to those of the beta-adrenergic agonist, ISO, and the phosphodiesterase inhibitor, isobutylmethylxanthine. The ability of ACh to reverse the positive inotropic effect of cholera toxin without preventing the accumulation of cyclic AMP may involve the prevention or reversal of cyclic AMP dependent phosphorylation. In this regard, reduction of Ca2+ influx through voltage-sensitive membrane channels may be an essential component of muscarinic inhibition. PMID- 6287869 TI - Brain potassium ion homeostasis, anoxia, and metabolic inhibition in turtles and rats. AB - Microelectrode measurements of tissue oxygen tension (PtO2) and extracellular potassium ion concentration ([K+]o) and dual wavelength spectrophotometric measurements of the reduction/oxidation state of cytochrome aa3 were used to compare the resistance of turtle and rat brain to anoxia in vivo. In both species, respiration with 100% N2 resulted in a decrease of tissue oxygen tension to near 0 mmHg and reduction of cytochrome aa3. However, N2 respiration resulted in only moderate elevation of [K+]o in turtle bran while [K+]o in rat brain was elevated to levels greater than 50 mM. In addition, N2 respiration in turtles had no effect on the rate of recovery of [K+]o, which was elevated by direct electrical stimulation of the brain. Electrocorticographic activity (ECoG) of the turtle brain was only moderately depressed during N2 respiration for up to 4 h whereas the ECoG of rat brain became isoelectric within 1 min. Inhibition of glycolysis with iodoacetate (IAA) resulted in rapid elevation of [K+]o in turtle brain during anoxia, but IAA had little effect on [K+]o during normoxia. These results indicate that the remarkable resistance of the diving turtle to anoxia does not result from continued provision of oxygen to the brain either by redistribution of systemic blood flow or from blood O2 storage. In addition, the turtle brain does not rely on cellular stores of high-energy compounds for maintenance of ionic homeostasis. We conclude that potassium ion homeostasis in the anoxic turtle brain must result from increased glycolytic ATP production and from decreased energy utilization. PMID- 6287868 TI - Ca2+-tolerant adult canine myocytes: preparation and response to anoxia/acidosis. AB - A procedure is described for the production of large numbers of Ca2+-tolerant adult canine ventricular myocytes. Gentle tissue dissociation is achieved in Spinner flasks using 320 mosM enzyme buffer containing collagenase hyaluronidase, and trypsin in the presence of millimolar levels of Ca2+. The technique allows for the complete removal of erythrocytes, the gentle removal of closely adhering nonmuscle cells (less than 3% contamination), and the selective removal of damaged from nondamaged myocytes. Total myocyte yields averaged 4-6 x 10(6)/g wet wt; 61.0 +/- 6.5% of the cells have rod-shaped morphology and are "viable" based on trypan blue exclusion. Only 3.9 +/- 1.1% of the rod-shaped cells beat spontaneously when challenged with 2 mM Ca2+. Exposure to 2 mM Ca2+ at 37 degrees C results in minimal loss of viability (2.1 +/- 1.3%/h) based on both trypan blue uptake and creatine kinase release. Simulation of cellular (i.e., membrane) injury in vitro is performed by the separate application of the perturbations of anoxia, acidosis, and low glucose to the canine myocyte suspensions; the data suggest that these myocytes are affected independently by these perturbations and are in good agreement with the results obtained by others using the isolated rat myocyte system. PMID- 6287870 TI - Early responses to hemorrhage in the conscious rat: effects of corticosterone. AB - There is evidence for a physiological role of the adrenal cortex in the early responses to limb ischemia in the rat. Trilostane, which inhibits steroid production and prevents the usual rise in corticosterone concentration, impairs compensatory fluid movement during the 3 h after removal of bilateral hindlimb tourniquets and truncates the accompanying hyperglycemia. We have now studied whether altering the corticosterone concentration has similar effects over a 3-h period after a 35% hemorrhage in the conscious rat. After hemorrhage there was only a small rise in plasma glucose concentration, which was unaffected by inhibition of the adrenocortical response with trilostane or its prolongation with adrenocorticotrophic hormone. However, if hindlimb tourniquets were applied 4 h beforehand, the hyperglycemia after hemorrhage was as large as after tourniquet removal and was similarly curtailed by trilostane. Compensatory fluid movement, in contrast, was unaffected by any of the alterations in corticosterone concentration, with or without tourniquets. Thus the method of producing fluid loss is critical in determining whether glucocorticoids play a role in compensation but not in maintaining hyperglycemia after injury. PMID- 6287871 TI - Persistence of urinary marijuana levels after supervised abstinence. AB - The authors present a case report of the presence of urinary cannabinoids during 21 days of supervised abstinence from chronic marijuana use and provide data on 6 similar cases. They discuss the theoretical implications of the persistence of cannabinoids. PMID- 6287872 TI - Malignant mesothelioma: ultrastructural distinction from adenocarcinoma. AB - Mesotheliomas and metastatic adenocarcinomas involving the pleura are frequently difficult to distinguish by light-microscopic and histochemical methods. In a double-blind study, we have compared ultrastructural features of 10 mesotheliomas of epithelial type and 10 adenocarcinomas from the lung, breast, and upper GI tract, i.e., sites known to give rise to metastases which mimic mesothelioma. Mesotheliomas were observed to have a significantly greater microvillus length/diameter ratio (LDR) than adenocarcinomas (p less than 0.01) and more abundant intermediate filaments (p less than 0.001). Mesotheliomas had more complex microvilli than adenocarcinomas, whereas adenocarcinomas had rootlets (2/10 cases) and lamellar inclusion bodies (2/10 cases), both of which were absent in the mesotheliomas. This study provides quantitative and qualitative ultrastructural features of potential utility in the differential diagnosis of pleural mesotheliomas and adenocarcinomas. PMID- 6287874 TI - Recurrent and metastatic cystosarcoma phyllodes. AB - During a 30 year period (1950 to 1980), 42 patients with cystosarcoma phyllodes were seen at the Mayo Clinic. Ten patients (24 percent) had recurrence of metastasis. The malignant or metastatic potential could not be identified or predicted by histologically evaluating tumor contour, stromal atypia, or mitotic activity. Spread was chiefly by a hematogenous route, and no patient with metastasis survived. Five of the 10 patients died from their disease 2 to 7 years after treatment. Simple mastectomy appears to be the surgical treatment of choice. PMID- 6287873 TI - A black hepatocellular carcinoma with Dubin-Johnson-like pigment and Mallory bodies: a histochemical and ultrastructural study. AB - A hepatocellular carcinoma which was predominately black was surgically excised from a noncirrhotic, asymptomatic 62-year-old white man. Brown-black, pigment granules, found only in the tumor cells, were histochemically and ultrastructurally identical to the hepatocellular pigment found in Dubin-Johnson syndrome. The latter pigment is thought to accumulate as a consequence of a genetically determined abnormality in the excretion of catecholamines and related substances. It is postulated that the pigment formation in this tumor developed via a similar, though epigenetic, mechanism. This occurrence has not been previously described. Unusual PAS-negative, globular cytoplasmic inclusions were also found in the tumor cells and these proved to be Mallory bodies by electron microscopy. PMID- 6287875 TI - Hepatopathy following irradiation and chemotherapy for Wilms' tumor. AB - Two children being treated with combination chemotherapy and irradiation for localized, rightsided Wilms' tumor developed sudden enlargement of the liver with defects on liver scintigram resembling liver metastases. One child also developed pancytopenia. When chemotherapy was temporarily withheld in both children, hepatomegaly and scintigram abnormalities resolved. The planned courses of chemotherapy were subsequently completed without complications. The clinical course in our patients is compared to previously published experiences. Awareness of this complication could prevent the mistaken diagnosis of metastatic disease and emphasizes the care necessary when administering cytotoxic drugs to children receiving irradiation to all or a portion of the liver. PMID- 6287877 TI - A new multifunction nerve stimulator. PMID- 6287876 TI - Cytomegalovirus infection of the skin. AB - A case of fatal generalized cytomegalovirus infection with particular attention to lesions involving the skin is reported. Characteristic cytomegalovirus inclusion bodies were present within nuclei of endothelial cells in the dermis beneath ulcerations. Skin biopsy may be useful in the diagnosis of disseminated infections by cytomegalovirus. PMID- 6287878 TI - The facile generation of covalently closed, circular DNAs with defined negative superhelical densities. PMID- 6287879 TI - A simple and efficient hybridization technique for quantitating virus-specific ribonucleates in polyoma virus-infected cells. PMID- 6287880 TI - Adenosine 5'-monophosphate-removing system in fructose-1,6-bisphosphatase assay mixture: a new approach. PMID- 6287881 TI - A simple spectrophotometric method for determining proton release from yeast cells. PMID- 6287882 TI - A practically sensitive radioimmunoassay for cyclic CMP by 2'-O-acetylation. PMID- 6287883 TI - The use of high-energy microwave irradiation to inactivate mitochondrial enzymes. PMID- 6287884 TI - Continuity of intracellular channels with extracellular space in adipose tissue and liver: demonstrated with tannic acid and lanthanum. AB - Tannic acid was used to demonstrate continuity of intracellular channels with extracellular space in white adipose tissue of adult rats, brown adipose tissue of suckling rats, and liver of diabetic rats. Electron-opaque material resulting from treatment of glutaraldehyde-fixed tissue with tannic acid was found in extracellular space, invaginations of cell surfaces, vesicles, and intracellular channels. Electron-opaque material was present in channels that surrounded lipid droplets in both white and brown adipocytes and in hepatocytes. The small distance between the lumen of marked channels and lipid droplets in adipocytes indicates that a monolayered structure, perhaps a leaflet of membrane lining the channel. Similar findings were obtained in brown adipose tissue using lanthanum instead of tannic acid to mark intracellular channels continuous with extracellular space. Since endoplasmic reticulum is the primary site of triacylglycerol synthesis in adipocytes, marked channels near lipid droplets may be elements of endoplasmic reticulum. Some of the channels marked with tannic acid in hepatocytes contained lipoprotein particles, whereas others were located, in relation to mitochondria and lipid droplets, in the same sites as endoplasmic reticulum in untreated tissue. This indicates that some of the channels marked with tannic acid in hepatocytes are endoplasmic reticulum. Presence of electron opaque material in intracellular channels and vesicles, but not in cytoplasm, of treated tissue indicates the channels and vesicles were open to extracellular space during treatment with tannic acid or lanthanum and, furthermore, that their membranes were continuous with plasma membrane. PMID- 6287885 TI - Presence of luteinizing hormone receptor binding inhibitor (LH-RBI) in ovine testis. AB - Luteinizing hormone receptor binding inhibitor (LH-RBI has been isolated from the aqueous extracts of ram testis using Sephadex column chromatography. Sephadex G 75 fraction I was found to inhibit the binding of I125 LH to rat testis receptors. Further purification of Sephadex G-75-I fraction on G-200 column gave four fractions (I-IV), the maximum inhibitory activity to inhibit I125 LH binding to rat testis receptor was associated with the fraction I only. Fraction II gave marginal inhibition only, whereas fractions III and IV did not have any inhibitory effect on I125 LH binding to rat testis receptors. Our findings suggest that LH-RBI isolated from ram testis is a protein having molecular weight more than 10,000 Daltons. PMID- 6287886 TI - Free radical formation in vivo and hepatotoxicity due to anesthesia with halothane. AB - In vivo studies were undertaken to determine whether free radical formation in the liver during administration of various halogenated anesthetics is associated with hepatotoxicity of these agents in an animal model. In addition to the anesthetics halothane, enflurane, and isoflurane, carbon tetrachloride was studied as an example of a hepatotoxic halogenated compound acting by a free radical mechanism. Free radicals were trapped in vivo during anesthesia as stable adducts using the spin trap, alpha-phenyl-t-butyl nitrone. These adducts were extracted from the liver and studied by electron spin resonance spectrometry. Free radicals were detected after administration of halothane or carbon tetrachloride, compounds which were hepatotoxic under the conditions of the experiment, but were not found after anesthesia induced with enflurane or isoflurane, anesthetics which were not hepatotoxic under identical conditions. The free radical trapped after alpha-phenyl-t-butyl nitrone treatment of halothane-anesthetized rats appeared to be a metabolic intermediate of halothane. PMID- 6287887 TI - Pharmacodynamics and pharmacokinetics of metocurine in humans: comparison to d tubocurarine. PMID- 6287888 TI - The hemodynamic effects of calcium channel blocking agents: a brief review. PMID- 6287889 TI - Calcium antagonists in the treatment of individuals with ischemic heart disease. AB - The so-called calcium antagonists--verapamil, nifedipine, diltiazem, and perhexiline--are pharmacologic agents that will have a dramatic impact on the medical therapy of individuals with ischemic heart disease. They have been used extensively in Europe and Japan for over 15 years, and more recent studies from this country have demonstrated their efficacy in the treatment of patients with Prinzmetal's "variant" angina pectoris, stable (exertional) angina pectoris, and unstable angina pectoris. In addition, they appear effective in protecting ischemic myocardium both in patients with acute myocardial infarction as well as in those undergoing cardiopulmonary bypass. The purpose of this review is twofold: (a) to describe the mechanism of action and pharmacology of these agents and (b) to delineate their role in the treatment of individuals with ischemic heart disease. PMID- 6287891 TI - Fine needle aspiration cytopathology of bronchial carcinoid tumors: an analytical study of the cells. AB - While frequently considered to originate in a common stem-cell of neuroendocrine origin, bronchial carcinoid tumors and small-cell anaplastic carcinomas differ significantly in their biologic potential and treatment. Patient management is often dependent on the diagnostic specificity of a pulmonary fine needle aspiration specimen. This study evaluated the cellular features in fine needle aspiration samples from seven bronchial carcinoid tumors and four small-cell anaplastic carcinomas of the "intermediate" type. Planimetric measurements were performed on tracings of 1,100 cells. An analysis of specific cytoplasmic and nuclear features was also obtained on 2,200 cells. Significant quantitative and qualitative differences in the cytomorphology of the cells derived from bronchial carcinoid and small-cell anaplastic neoplasms were obtained, clearly demonstrating that differentiation of these neoplasms is possible on a cytopathologic basis in fine needle aspiration samples. PMID- 6287890 TI - Management of arrhythmias with "calcium antagonists". PMID- 6287892 TI - Flow cytometry of lung carcinoma: a comparison of DNA stemline and cell cycle distribution with histology. AB - Flow cytometry studies of the DNA distribution of 33 lung tumors were carried out. All of the carcinomas (32 cases) had aneuploid DNA modal values, ranging from 2.15c to 5.05c; in the single case of carcinoid studied, the tumor cells were diploid. DNA ploidy levels tended to be higher for epidermoid than adenocarcinoma; they were the same in lymph node metastases as in the primary tumor. Cell cycle distributions calculated from the tumor cell DNA values showed considerable variation, ranging from 9% to 58% for the S phase and from less than 1% to 29% for the G2M phase. Whether these variations have clinical significance is not known at this time. PMID- 6287893 TI - Ketoconazole blocks adrenal steroid synthesis. AB - Ketoconazole, a broad-spectrum, antifungal drug that is administered orally, has been shown to inhibit sterol synthesis in fungi. When gynecomastia developed in some patients taking this drug, we investigated the effects of ketoconazole on steroid synthesis in humans and in isolated adrenal cells from rats. In healthy humans, the cortisol response to adrenocorticotropic hormone was significantly blunted 4 hours after a 400-mg or 600-mg dose. The inhibition persisted for up to 8 hours and was absent by 16 hours. This finding indicated that adrenal androgen response was reduced. Easily achieved therapeutic concentrations of ketoconazole virtually eliminated corticosterone production by isolated adrenal cells from rats. Although ketoconazole at currently used doses has never been documented to cause clinical hypoadrenalism, caution is urged in high- or multiple-dose trials. The drug may prove useful as an agent to block steroid synthesis. PMID- 6287894 TI - [Electron microscopy studies of autopsy lung specimens from 26 children with fatal acute respiratory diseases]. PMID- 6287895 TI - [Isolation and identification of viral causative agents in an episode of infant mortality occurring in Naples from October 1978 to April 1979]. PMID- 6287896 TI - [Neuropsychic complications of disulfiram therapy. Apropos of a case]. PMID- 6287897 TI - Herpetic whitlow and ocular infection. AB - Herpes simplex virus can involve the mouth, nose, or skin in addition to the eye. A very rare form of herpes simplex involves the fingertips (herpetic whitlow) and can be a source of diagnostic confusion. Herpes simplex can be spread by direct contact from any of these sites to the eye by cross-infection or autoinoculation. The ophthalmologist should be aware of the nonocular forms of herpes simplex. PMID- 6287898 TI - Extended transantral approach to pterygomaxillary tumors. PMID- 6287900 TI - The hemostatic effect of conjugated water-soluble estrogens in rats. AB - Estrogenic steroids have been used as hemostatic agents in various surgical procedures in humans. Their use to reduce bleeding has been based mainly on clinical observation. The present study was carried out to test experimentally whether intravenously administered Premarin reduces capillary bleeding. Skin graft donor sites in rats served as the standard capillary bleeding source in this controlled double-blind experiment, in which Premarin was found to appreciably reduce capillary bleeding. PMID- 6287899 TI - Pathology consultation. Metastatic patterns of salivary gland neoplasms. AB - Long-term follow-up of salivary gland carcinomas allows a better evaluation of their biologic malignancy than the traditional five-year period. Metastases (distant and local) are possible over the entire lifetime of a patient and are dependent upon histologic grade, persistence of neoplasm and clinical stage. Distant metastases to bone and lungs are manifested by nearly every carcinoma. Metastases to regional lymph nodes vary according to histologic type and it appears that the adenoid cystic carcinoma has the lowest incidence of that event. PMID- 6287901 TI - Bilateral superomedial thigh flaps for primary reconstruction of scrotum and vulva. AB - The use of bilateral superomedial thigh flaps for primary reconstruction of scrotum and vulva is presented. These flaps have proved to be reliable, ensuring good healing and providing almost normal sensation. Further indirect evidence that these are arterial flaps is given. PMID- 6287902 TI - Platelet aggregation inhibitors and cancer metastasis. PMID- 6287903 TI - [Biochemical data on C1 intrinsic activation (author's transl)]. AB - C1 activation can be triggered by immune complexes and various effectors such as extrinsic proteases, bacterial or viral membranes, polyanions and polysaccharides. The basic mechanism of activation involves a limited proteolytic cleavage of C1r, with the generation of a proteolytic activity. Highly purified proenzymic C1r was obtained in high yield from human plasma by an indirect affinity chromatography step. Activation of isolated C1r in a fluid phase proceeded according to two distinct coexisting mechanisms: 1) an autocatalytic intradimer activation mediated by the pro-site of non-activated C1r; 2) an autocatalytic interdimer proteolysis triggered by nascent activated C1r formed in the course of the first reaction. DFP and C1-inhibitor did not have any effect on the first mechanism but were able to block the second mechanism. C1 activation is discussed in the light of recent results obtained by others from electron microscopy, and a tentative model is proposed. PMID- 6287904 TI - Antibody-independent activation of C1, the first component of complement. PMID- 6287905 TI - Cyclic AMP mediated morphological changes in a hybrid cell line. AB - A hybrid cell line (PCM 3) obtained from a fusion between Chinese hamster fibroblasts and a mamalignant lymphoma has the unusual property of converting from a monolayer of flat spindle-shaped cells to one of spherical cells on treatment with agents which elevate intracellular cAMP. This dose dependent changes is not accompanied by a loss of adhesion and is enhanced by phosphodiesterase inhibitors. The morphological change has been found to be energy dependent and is similar to that induced by cytochalasin B, although the changes has been found to be energy dependent and is similar to that induced by cytochalasin B, although the changes in plasma membrane proteins are not of a similar nature. Furthermore, it has been shown that specific serum factors can inhibit this cAMP induced cell rounding. PMID- 6287906 TI - Representation of mRNA sequences in normal and SV40 transformed human diploid fibroblasts. AB - The representation of mRNA sequences in normal and SV40-transformed WI-38 human diploid fibroblasts was examined. Variations in the hybridization of polysomal RNAs to 3H-labeled DNAs complementary to poly A+ mRNA of normal and SV40 transformed cells suggest differences in the mRNA populations. a comparison of the representation of histone RNA sequences of normal and SV40-transformed human diploid cells did not reveal significant differences. PMID- 6287907 TI - New technologies to combat malignant tumours of the brain. AB - 1. The primary problem in an effective treatment of a glioblastoma is the prevention of a recurrence. 2. For that purpose were the following therapeutical procedures undertaken: (a) Temporary implantation of radio cobalt in the brain itself (1957): (b) Clostridium butyricum M 55 was used to render the centre of the tumour fluid (1967): (c) Podophyllin was used to destroy the border of the tumour (1980); (d) The CO2 Laser beam (1975); (e) The electromagnetic heat induction deep in the brain (1973-1978). 3. In order to make the operation and postoperative phase safer for the patient, the following precautions were drawn upon or employed: (a) Hyperbaric oxygenisation in the pressure chamber (1971); (b) The anti-G-suit (1974); (c) the computer controlled automatic infusion pump (1980), and (d) the telemetric measurement of intra-cranial pressure (1975). 4. Apart from the pressure chamber, the mentioned devices were all supervised and developed in the department of the author. 5. The first successful means in the prevention of the recurrence of a glioblastoma multiform seems to be the telethermic method mentioned in 2 (e) above. PMID- 6287908 TI - Adenoid cystic carcinoma: a third type of human salivary gland neoplasms characterized cytogenetically by reciprocal translocations. AB - Using G-banding technique, the chromosomes were studied in four consecutive preparations from cultured material from a human adenoid cystic carcinoma of the submandibular gland. The results indicated that the carcinoma had originated with a normal diploid stemline, and that reciprocal translocations played a predominant role in development of an abnormal stemline. The propensity of developing variant cells, as well as an abnormal stemline, characterized by various types of reciprocal translocations, was found to be a property shared by two other types of salivary gland neoplasms studied by banding techniques, namely the pleomorphic adenoma and the acinic cell tumour. The tissue of origin is suggested to be the crucial determinant responsible for this important cytogenetical similarity between different types of salivary gland tumours. PMID- 6287910 TI - Periodic paralysis and the sodium-potassium pump. AB - Analysis of the pathophysiology of hypokalemic paralysis, as it occurs in barium poisoning, chronic potassium deficiency, and thyrotoxicosis, suggests that these disorders may have a similar mechanism. An increased ratio of muscle sodium permeability to potassium permeability reduces the ionic diffusion potential, while the resting membrane potential is sustained by an increase of Na-K pump electrogenesis. The result is that potassium entry (the sum of active and passive influx) exceeds potassium efflux; this causes a large shift of extracellular potassium into muscle until the Na-K pump turns off, leading to depolarization and paralysis. The primary defect in familial hypokalemic periodic paralysis, as in the example of barium poisoning, may be a marked reduction of muscle permeability to potassium. PMID- 6287909 TI - Morphology of spontaneous and induced tumors in the bronchiolo-alveolar region of F344 rats. AB - The morphology of spontaneous and chemically-induced metastasizing carcinomas and adenomas in the bronchiolo-alveolar region of F-344 rats was studied. Histologically, the tumors were tubulo-papillary. Ultra-structurally, they consisted of cells which formed and secreted osmiophilic lamellated inclusion bodies, a marker of alveolar type II cells. Mitotic tumor cells also demonstrated such bodies. No cells of bronchial or bronchiolar origin were found in the tumors. We conclude that in F344 rats, lung tumors located in the bronchioloalveolar region consist of alveolar type II cells exclusively and are, therefore, alveolar cell adenomas and carcinomas, respectively. PMID- 6287911 TI - A newly recognized congenital myasthenic syndrome attributed to a prolonged open time of the acetylcholine-induced ion channel. AB - Five familial cases (in two families) and one sporadic case of a new congenital myasthenic syndrome were investigated. Symptoms arise in infancy or later life. Typically, one finds selective involvement of cervical, scapular, and finger extensor muscles, ophthalmoparesis, and variable involvement of other muscles. There is a repetitive muscle action potential to single nerve stimulus in all muscles and a decremental response at 2 to 3 Hz stimulation in clinical affected muscles. Microelectrode studies reveal markedly prolonged end-plate potential (epp), miniature end-plate potential (mepp), and miniature end-plate current; normal quantum content of the epp; and a smaller than normal or low-normal mepp amplitude. Light microscopy demonstrates predominance of type I fibers, small groups of atrophic fibers, tubular aggregates and vacuoles near end-plates, abnormal end-plate configuration, and nonspecific myopathic changes. Abundant acetylcholinesterase activity is present at all end-plates, and the activity and kinetic properties of this enzyme in muscle are normal. Calcium accumulated at the end-plate in one patient. Quantitative electron microscopy shows decrease in the size of nerve terminals, increase in the density of synaptic vesicles, and reduction in the length of postsynaptic membranes. There is focal degeneration of junctional folds with corresponding loss of acetylcholine receptor, most marked in cases with the lowest mepp amplitude. There are no immune complexes at the end plate. Fiber regions near end-plates display dilation, proliferation, and degeneration of the sarcoplasmic reticulum; nuclear, mitochondrial, and myofibrillar degeneration; and vacuoles resembling those found in periodic paralysis. A prolonged open time of the acetylcholine-induced ion channel is considered to be the basic abnormality and may account for the physiological, morphological, and clinical alterations. PMID- 6287912 TI - Inclusion-body myositis: clinicopathological studies and isolation of an adenovirus type 2 from muscle biopsy specimen. PMID- 6287913 TI - Hind limb somatosensory evoked potentials in the monkey: the effects of distal axonopathy. AB - Computer-averaged short-latency somatosensory evoked potentials (SLSEP) to unilateral stimulation of the peroneal nerve were recorded from surface electrodes overlying the peripheral nerve, cauda equina, spinal cord, brainstem, and contralateral sensorimotor region. Seven monkeys were studied under normal conditions and at various stages of distal axonopathy induced by systematic acrylamide intoxication. With the use of a noncephalic reference, a series of five small-amplitude positive components were identified that precede the initial cortical activity. On the basis of timing and topography of the components, the source of the first one, SLSPEP1, was localized to the lumbar dorsal root fibers and that of the second, SLSEP2, to the ascending spinal tracts, principally gracile fasciculus. Bipolar recordings of the SLSEP2 overlying the rostral extreme of the cervical spinal cord provided a sensitive measure of the onset of distal axonopathy. Changes in the timing of this component antedated both abnormalities of spinal or peripheral nerve conduction and behavioral signs of intoxication. The initial alteration was specific to stimulation of the hind limb and was associated with early morphological change limited to the terminal and preterminal portions of the long axons in the gracile fasciculus. PMID- 6287914 TI - Proton electrochemical gradients and energy-transduction processes. PMID- 6287915 TI - Enzymology of oxygen. PMID- 6287916 TI - Enzymes of the renin-angiotensin system and their inhibitors. PMID- 6287917 TI - Mechanisms of intracellular protein breakdown. PMID- 6287918 TI - Magnetic resonance studies of active sites in enzymic complexes. PMID- 6287919 TI - Specific intermediates in the folding reactions of small proteins and the mechanism of protein folding. PMID- 6287920 TI - Carbohydrate-specific receptors of the liver. PMID- 6287921 TI - Bacteriorhodopsin and related pigments of halobacteria. PMID- 6287922 TI - DNA repair enzymes. PMID- 6287923 TI - Molecular mechanisms in genetic recombination. PMID- 6287924 TI - The biology and mechanism of action of nerve growth factor. PMID- 6287925 TI - Chromatin. PMID- 6287926 TI - [Clinico-laboratory basis for the endolymphatic use of beta-lactam antibiotics in pulmonology]. AB - Clinical efficacy and effect of cefuroxime, claforan and pentrexyl used endolymphatically were studied in 85 patients with acute abscess forming and persisting pneumonia. Previous routine antibiotic therapy in these patients was little effective. Administration of the antibiotics into the peripheral lymph nodes provided blocking of the lymphagenic pathway for the infection due to high levels in the lymphatic system. Endolymphatic use of cefuroxime and claforan resulted in a significant improvement of the functions of the T- and B-immunity systems and the indices of natural resistance. The levels of the autoimmune reactions and sensitization to the bacterial antigens decreased. Endolymphatic use of cefuroxime and claforan once every 3 days provided recovery of 9 2.8 per cent of the patients, the treatment periods being decreased 2.5--3 times. Intravenous administration of the drugs according to the routine schemes, endolymphatic use of pentrexyl (5 g once every 3 days) and endolymphatic administration of cefuroxime in a single dose followed by intravenous therapy was less effective. The efficacy of pentrexyl increased, when it was used endolymphatically in combination with lysozyme. Endolymphatic use of claforan in doses of 2--3 g once every 3 days (3--4 infusions during the treatment course) was most effective. PMID- 6287927 TI - Molecular epidemiology of beta-lactamase-specifying plasmids of Haemophilus ducreyi. AB - We have studied the genetic basis of beta-lactamase production in eight strains of Haemophilus ducreyi isolated in diverse areas of the world. Beta-lactamase production in all strains was mediated by plasmids having a molecular mass of either 5.7 or 7.0 megadaltons. Plasmids of 5.7 megadaltons were shown to carry the entire sequence of pFA7, the beta-lactamase specifying plasmid found in isolates of Neisseria gonorrhoeae epidemiologically linked to West Africa. Plasmids of 7.0 megadaltons were shown to carry the entire sequence of pFA3, the beta-lactamase specifying plasmid found in Far Eastern isolates of N. gonorrhoeae. Both groups of H. ducreyi plasmids were shown to carry physically complete and functional TnA sequences. Thus we have identified two types of H. ducreyi beta-lactamase plasmid which are identical to the two types of N. gonorrhoeae beta-lactamase plasmid, except that they carry complete TnA sequences. PMID- 6287928 TI - Prophylactic activity of intranasal enviroxime against experimentally induced rhinovirus type 39 infection. AB - Intranasal administration of enviroxime by aerosol spray was associated with drug levels in nasal secretions that 1 h later averaged 750-fold higher than those inhibitory for rhinoviruses in vitro (0.2 microgram/ml). However, administration of intranasal enviroxime (one spray per nostril, five times per day) to susceptible volunteers, beginning 1 day before and continuing for 4 days after virus exposure, did not significantly reduce infection or illness due to experimentally induced rhinovirus type 39 infection. The combined results of two separate trials yielded an infection rate of 100% for 21 placebo-treated and 89% for 19 enviroxime-treated subjects. Approximately one-half of the volunteers in each group had seroconversion to the challenge virus. Overall, 52% of the placebo treated and 53% of the enviroxime-treated subjects developed colds. No significant differences in symptom scores, nasal mucus weights, or numbers of nasal tissues used were observed between the two groups. Two-thirds of the enviroxime-treated volunteers noted intranasal irritation immediately after sprays, as compared with only one-third of the placebo-treated subjects. PMID- 6287930 TI - Effect of ketoconazole on isolated mitochondria from Candida albicans. AB - Ketoconazole, an oral antimycotic imidazole drug, blocked the transport of electrons in the respiratory chain of Candida albicans under aerobic conditions with different substrates, such as NADH and succinate. This effect was a nonspecific inhibition of NADH oxidases and succinate oxidases. The addition of ketoconazole to C. albicans mitochondria without a substrate resulted in strong reduction of cytochrome a3, as revealed by difference spectra (reduced versus oxidized). This indicated that there was a specific interaction between ketoconazole and cytochrome c oxidase. A spectrophotometric analysis confirmed that the cytochrome oxidases other than cytochrome c oxidase were not inhibited because subsequent addition of any substrate caused an increased level of reduction of all of the other respiratory chain components compared with the control. Consequently, our data strongly suggested that the primary site of ketoconazole inhibition on isolated mitochondria from C. albicans is the most distal portion of the respiratory chain. PMID- 6287929 TI - Primary site of action of ketoconazole on Candida albicans. AB - Ketoconazole, an antifungal drug, completely inhibited the growth of Candida albicans 7N at concentrations of greater than or equal to 50 microgram/ml (94 microM). However, ketoconazole incompletely inhibited the growth of this opportunistic yeast at concentrations of 25 to 0.2 microgram/ml (47 to 0.4 microM). At these lower concentrations, 2,3,5-triphenyl tetrazolium chloride, an electron acceptor, was reduced by several strains of C. albicans. This effect resulted in red coloration of colonies. Concomitantly, this phenomenon was not antagonized in the presence of ergosterol. Furthermore, neither ketoconazole nor antimycin A inhibited the growth of C. albicans under anaerobic conditions, as revealed by a paper disk method. Ketoconazole at the concentrations stated above inhibited endogenous and exogenous respiration immediately after it was added to a system containing log phase C. albicans cells, as determined polarographically. At the same time, ketoconazole inhibited the activity of NADH oxidase at the mitochondrial level. In contrast, higher concentrations of ketoconazole (greater than 100 microM) were required to inhibit the activity of succinate oxidase from rat liver mitochondria. In addition, concentrations of ketoconazole greater than 100 microM were required to impair the uptake of labeled leucine and adenine and, subsequently, the incorporation of the former into protein and the latter into DNA and RNA in intact cells. On the other hand, ketoconazole at concentrations of 10, 1.0, and 0.4 microM had no effect on either membrane permeability or macromolecular synthesis. PMID- 6287931 TI - Pharmacokinetics and tolerance of ceftriaxone in humans after single-dose intramuscular administration in water and lidocaine diluents. AB - The effects of 1% lidocaine as a diluent on the pharmacokinetics and tolerance of ceftriaxone administered intramuscularly were investigated in 12 adult volunteers. Each subject received two 0.5-g doses of ceftriaxone (one in water and the other in 1% lidocaine) at least 1 week apart in a randomized crossover fashion. Plasma and urine samples were collected serially and assayed for ceftriaxone content by high-performance liquid chromatography. The mean peak plasma concentration, time to attain the peak, area under the plasma curve from time zero to infinity, and elimination half-life were 45 micrograms/ml, 2.5 h, 578 micrograms . h/ml, and 7.1 h, respectively, after intramuscular administration of ceftriaxone in water diluent. The corresponding mean values in 1% lidocaine diluent were 42 micrograms/ml, 3 h, 577 micrograms . h/ml, and 7.0 h. The pharmacokinetic data suggested that 1% lidocaine does not alter either the elimination parameters or the bioavailability of intramuscularly administered ceftriaxone. The intensity and frequency of pain at the injection site were reduced considerably by the coadministered lidocaine. PMID- 6287932 TI - Comparison of the activities of ceftriaxone and penicillin G against experimentally induced syphilis in rabbits. AB - The activity of ceftriaxone, a newly developed cephalosporin, against early cutaneous infections with Treponema pallidum in rabbits was compared with that of equimolar doses of penicillin G. Activity was related to the time required for cutaneous lesions to become dark-field negative, serological response, and the disappearance of T. pallidum from the popliteal lymph nodes. Both antibiotics were very effective in the treatment of syphilis in this animal model. The 50% curative dose for penicillin G was 0.8 mumol/kg (0.29 mg or 480 U/kg) and for ceftriaxone, it was 1.45 mumol/kg (0.96 mg/kg). Overall, ceftriaxone was slightly less effective than penicillin G was. Transmission and scanning electron microscopy studies of testicular aspirates obtained from rabbits treated with ceftriaxone revealed alterations in the treponeme surface which apparently resulted in changes in cell permeability and morphology. PMID- 6287933 TI - Comparative in vitro activities of cefmenoxime (SCE-1365) and newer cephalosporin derivatives of clinical utility. AB - The minimal inhibitory concentrations of cefmenoxime (SCE-1365), cefotaxime, cefoperazone, and moxalactam against various species of aerobic bacteria were determined. The activities of cefmenoxime, cefotaxime, and moxalactam were generally similar and slightly higher than the activity of cefoperazone. PMID- 6287934 TI - Activation of wheat chloroplast sedoheptulose bisphosphatase: a continuous spectrophotometric assay. PMID- 6287935 TI - Hyperoxia increases oxygen radical production in rat lung homogenates. PMID- 6287936 TI - Properties of a 5'-AMP specific nucleotidase which accumulates in one cell type during development of Dictyostelium discoideum. PMID- 6287937 TI - Human galactose-1-phosphate uridylyltrsferase: purification and comparison of the red blood cell and placental enzymes. PMID- 6287938 TI - Pyrazole and 4-methylpyrazole inhibit oxidation of ethanol and dimethyl sulfoxide by hydroxyl radicals generated from ascorbate, xanthine oxidase, and rat liver microsomes. PMID- 6287939 TI - The kinetics of benzo(a)pyrene anti-7,8-dihydrodiol 9, 10-epoxide formation from benzo(a)pyrene and regulatory membrane effects. PMID- 6287940 TI - Electron paramagnetic resonance study of nitrosylprotoheme dimethyl ester complexes with aliphatic nitrogenous bases: characterization of the axial ligand trans to the nitrosyl group in nitrosylhemoproteins. PMID- 6287941 TI - Solubilization and assay of an hepatic receptor for the haptoglobin--hemoglobin complex. PMID- 6287943 TI - A new toxin isolated from Xanthomonas malvacearum which inhibits mitochondrial ATP--ADP translocase. PMID- 6287942 TI - Kinetic indication for multiple sites of ubiquinol-1 interaction in ubiquinol cytochrome c reductase in bovine heart mitochondria. PMID- 6287944 TI - Ultrastructural studies of necrolytic migratory erythema. AB - Necrolytic migratory erythema is a disorder highly suggestive of glucagonoma syndrome. We carried out histologic and electron microscopic studies of the skin lesions in a 57-year-old woman with the glucagonoma syndrome. Light microscopic studies revealed hyperkeratosis, parakeratosis, formation of clefts in the upper epidermis, and scattered dyskeratotic cells. Ultrastructurally, the intercellular spaces were widened in the upper epidermis and desmosomes were reduced in number. The cytoplasm of affected cells showed vacuolar degeneration; the organelles were lysed or absent. There were scattered dark cells and dyskeratotic cells. These changes appear to represent a degenerative process of the keratinocytes. PMID- 6287945 TI - Renal pseudotumours caused by xanthogranulomatous pyelonephritis. PMID- 6287946 TI - Mercury toxicity as compared to adrenocorticotropin-induced physiological stress in the chicken. PMID- 6287947 TI - Effects on rat lung immunity by acute lung exposure to benzo(a)pyrene. AB - This study describes the effect of intratracheal instillation of benzo(a)pyrene (BaP) on immunological responses in the lung-associated lymph nodes, cervical lymph nodes, and spleen after deposition of 10(8) sheep red blood cells (SRBC) in the lung or peritoneal cavity of rats. An increased number of anti-SRBC antibody forming cells was observed in the lung-associated lymph nodes when rats were immunized simultaneously with BaP installation. A suppression in the number of anti-SRBC antibody-forming cells occurred when SRBC were given intratracheally 4 or 7 days after BaP. The effects of the BaP appeared to be on the function of the cells in the lung-associated lymph nodes rather than due to changes in the exposed lung. BaP-induced changes in antigen handling or in regulatory populations of immune cells in the lung-associated lymph nodes may be responsible for the immune alterations observed. PMID- 6287948 TI - [Prolactin and cancer of the prostate. I. Prolactin: biochemistry, control of its secretion and functions. Its role in androgen metabolism and in prostate physiology]. PMID- 6287949 TI - Surgical management of the ectopic ACTH syndrome. AB - Most patients with extra-pituitary ACTH-secreting tumors die from carcinoma, but a few can benefit from operation. Of 96 patients with Cushing's syndrome, 11 probably had such tumors. There were three modes of presentation: (1) three had malignant tumors with visceral metastases initially. One (bronchial carcinoid) died without operation. Two with carcinoma (thyroid medullary and islet-cell) underwent adrenalectomy with remission, but died soon. (2) three had apparently benign tumors initially. One (appendicular carcinoid) underwent appendectomy and one (bronchial carcinoid with hilar node metastases) underwent lobectomy. Both had rapid remissions. The third (pheochromocytoma) died after resection of the tumor. (3) five patients had no obvious tumors and underwent adrenalectomy with remission. In one a benign bronchial carcinoid was removed later. Four others remain well, but without localizing signs of tumor. The main biochemical features in all were hypokalemic alkalosis and very high urinary excretion of free cortisol. Seven of the eight patients without visceral metastases are in remission from one to 15 years after operation. PMID- 6287950 TI - Adenylyl cyclase and protein carboxyl methylase in human spermatozoa. AB - Adenylyl cyclase (AC) and protein carboxyl methylase (PCM) activities in human ejaculated spermatozoa were measured in 31 men being evaluated for infertility at the Infertility Clinic. The spermatozoal Mn2+-sensitive AC activities ranging between zero and 1.4 nmol cyclic adenosine monophosphate (cAMP)/10(9) cells/min, was positively correlated with motility index [% motile sperm X degree (0-4)] (r = 0.72, rho less than 0.05). This confirms a role for cAMP in human sperm motility. PCM activities, ranging between zero and 160 pmoles (CH3OH/10(9) cells/min), were significantly correlated with AC activities (r = 0.86, p less than 0.001). However, PCM activities were not positively correlated with motility index (p = 0.065). PCM most probably is a factor involved in spermatozoal motility since two patients suffering necrospermia possessed spermatozoa deficient in both AC and PCM activities. PMID- 6287951 TI - Reversal by acetylsalicylic acid of the captopril-induced inhibition of angiotensin converting enzyme in the hindquarters of guinea-pig. AB - The conversion of angiotensin I to angiotensin II was studied in the isolated perfused hindquarters of guinea-pig. The relative enzyme activity was determined by vasoconstrictor response to the peptides and by the contraction of rat ascending colon superfused with the venous effluent. 45% of conversion of angiotensin I to angiotensin II was determined in this preparation as measured by vasoconstrictor responses. However, only 22% of conversion was detected in the venous return as measured in the rat colon. Captopril significantly inhibited angiotensin converting enzyme activity in this preparation. Acetylsalicylic acid, however, partially prevented the inhibitory effect of Captopril on converting enzyme activity in this vascular bed. The possible interactions of angiotensin converting enzyme activity and endogenous prostaglandins are discussed. PMID- 6287952 TI - Benzo [b]-promazines, chlorpromazine free radical and analino-N, N dimethylpropylamine analogs influence rat striatal DA-adenylate cyclase and calmodulin-phosphodiesterase. AB - A series of Benzo [b]-promazines and analino-N, N-dimethylpropylamine analogs and the free radical of chlorpromazine were compared to chlorpromazine and promazine in the rat striatum for their ability to inhibit either dopamine activated adenylate cyclase or calmodulin stimulation of a partially purified high Km cyclic AMP phosphodiesterase. Chlorpromazine and the corresponding free radical were generally the most potent inhibitors of the two enzyme preparations, however, Piperazino-Benzo [b]-promazine, 1-Oxo-Benzo [b]-promazine, N-38-76-3A and Benzo [b]-promazine were relatively effective inhibitors. To a lesser extent, tyrosine, N-57-77, Piperidino-Benzo [b]-promazine, Diethyl-Benzo [b]-promazine, promazine and 1-Oxo-Diethyl-Benzo [b]-promazine exerted varying degrees of antagonism of the two enzymes. In all instances the compounds inhibited dopamine sensitive adenylate cyclase to a greater extent than the calmodulin activated phosphodiesterase. PMID- 6287953 TI - Antiarrhythmic effects of cyclic guanosine 3' :5'-monophosphate and sodium nitroprusside on barium chloride arrhythmias in rabbits. AB - The hypothesis that c GMP and sodium nitroprusside (NP), an activator of guanylate cyclase which elevates levels of c GMP, have antiarrhythmic activity was tested in the barium chloride (BaCl2) model of arrhythmias. Electrocardiograms were recorded continuously on tape in unanesthetized New Zealand white rabbits weighing approximately 2.0 kg. BaCl2, 4 mg/kg i.v. bolus, induced frequent ventricular ectopic beats. These ventricular arrhythmias were abolished by 8-bromo-c GMP, 5 mg/kg, injected into the left ventricle (5/6 rabbits), NP 25.0 microgram/kg/min i.v. (6/6), NP 10 micrograms/kg/min i.v. (3/6), and markedly reduced in frequency by NP 10 micrograms/kg/min (3/6). In temporal association with NP, 4 fold increases in c GMP levels in blood and significant increases in myocardial GMP were found compared to control animals (n = 6). In this model, c GMP and NP have significant antiarrhythmic properties. NP effect may be mediated by alterations in c GMP metabolism. PMID- 6287954 TI - [Urea cycle enzymes in rats fed with corn]. AB - Activities of five urea cycle enzymes, carbamyl phosphate synthetase, ornithine transcarbamylase, argininosuccinate synthetase, argininosuccinase and arginase, were measured in liver of rats fed two types of diet: diet I: raw common corn and, diet II: made of a mixture of corn starch ("maicena"), "chuno (dried potato), sugar and fat. The activities obtained were compared with those of a group of control rats fed a balanced diet with an adequate protein content. Diets were administered after weaning, through an experimental period of 90 days, and enzymatic activities were measured at regular intervals during this time. Animals raised on diet I as well as those raised on diet II, showed low activity of the five urea cycle enzymes, but the decrease of enzymatic activity was more pronounced and appeared earlier in animals raised on diet II. Carbamyl phosphate synthetase and argininosuccinate synthetase, the two synthetases of this cycle, decreased earlier and in a greater degree than the other three enzymes in animals raised on these two types of diet. PMID- 6287955 TI - [Growth of the water hyacinth (Eichhornia crassipes (Mart) Solms Laubach) in the tropics]. AB - The growth and reproduction rates of water hyacinth (Eicchornia crassipes (Mart) Solms Laubach) in tropical areas, under natural and fertilized controlled conditions, were studied. The fertilizer used was swine fecal matter at three different levels: 1.25, 2.5 and 5%. The highest rate of growth was found with the 2.5% concentration, and this value was 49.16 g/m2/day on a dry-weight basis. Under natural conditions the rate of growth was 18.39 g/m2/day on a dry-weight basis. These results represent a yearly production in ton/ha among the highest reported in the world. PMID- 6287956 TI - [Changes in angiotensinogen and des-angiotensin I-angiotensinogen in man and rat upon inhibition of converting enzyme]. AB - The effects of converting enzyme inhibition on plasma renin substrate concentration were studied in man and rat. This study use new direct radioimmunoassays of angiotensinogen completing the classical enzymatic methods. In human investigation converting enzyme is inhibited after Captopril treatment. Our results demonstrated that resulting increase of plasma renin concentration enhanced the consumption of renin substrate as shown by the fall of angiotensinogen levels measured by indirect method. In the rat, we observed the same drop of renin substrate during MK421 administration. The fall of angiotensinogen levels, measured by indirect method, was not in agreement with results of direct radioimmunoassay. This discrepancy can be explained by the accumulation of des-angiotensin I-angiotensinogen in plasma. These modifications are potentiated by sodium depletion. PMID- 6287957 TI - [Prevention of the development of genetic hypertension by MK 421 in the SHR]. AB - MK 421, 25 mg/kg, administered daily by gavage to young spontaneously hypertensive rats (SHRs) from their 4th to 15th weeks of age almost completely inhibited genetic hypertension development. Since heart rate and cardiac index were not drug affected, prevention of genetic hypertension development was solely related to an early, potent and long-lasting limitation of the progressive increase in peripheral resistance which normally develops in SHRs during ageing. MK 421 slightly enhanced vascular responsiveness to exogenous norepinephrine and angiotensin II and reduced myocardial hypertrophy. Plasma renin concentration was increased. MK 421 slightly reduced body growth but did not affect fluid intake, urinary volume and urinary ADH, demonstrating that no water or salt retention developed. Finally, MK 421's preventive effects against genetic hypertension development persisted up to 10 weeks after treatment discontinuation. PMID- 6287958 TI - The human papillomaviruses. AB - The human papillomaviruses (HPVs) have long been recognized as the etiologic agents of a variety of benign cutaneous and mucosal proliferative entities. Currently, 11 distinct HPVs have been described, and each seems preferentially associated with a distinct clinical entity. In animal species, some papillomaviruses (PVs) are associated with lesions that can become malignant. What role specific HPVs may play in cutaneous or mucosal malignant squamous cell neoplasms is under investigation in a number of different laboratories. There is a subgroup of PVs that can malignantly transform fibroblasts of heterologous species. This system provides a model for studying the malignant potential of the PVs and for determining the cellular and viral events associated with PV-induced malignant transformation. A unique characteristic of this system is that the HPV genome exists exclusively nonintegrated as a DNA plasmid in transformed cells. PMID- 6287959 TI - Unusual inclusions in plasmacytoid cells. Their occurrence in a patient with Waldenstrom's macroglobulinemia. AB - The bone marrow from a patient with Waldenstrom's macroglobulinemia was studied, and histologic, immunopathologic, and ultrastructural features correlated. The infiltrating plasmacytoid lymphocytes contained PAS-negative cytoplasmic inclusions, which were shown to contain a specific immunoglobulin that matched the patients's circulating paraprotein. Electron microscopic studies revealed abundant dense fibrillar and globular material within dilated cisternae of rough endoplasmic reticulum. This material was morphologically similar to, but quantitatively different from, an endoplasmic reticulum-associated structure previously described in several patients with chronic lymphocytic leukemia. The differences between these fibrillar inclusions and Russell bodies are stressed. PMID- 6287961 TI - Immunohistochemical acid phosphatase level and tumor grade in prostatic carcinoma. AB - An immunoperoxidase technique to detect prostatic-specific acid phosphatase (PSAP) was used on specimens from 98 cases of prostatic carcinoma that were graded by both the Gleason and the Mostofi systems, to see if tumor grade correlated with amount of PSAP seen in tissue. Most tumors showed strong, diffuse cytoplasmic staining; no significant difference was seen among the various grades. Other than focal, weak staining of renal tubular epithelium, the antibody to PSAP gave uniformly negative results with a variety of normal and neoplastic tissues. In light of the great sensitivity and specificity of this technique, it potential applications include diagnosis of poorly differentiated prostatic malignant neoplasms, whether primary or metastatic. PMID- 6287960 TI - Multiple granular cel tumors of the gallbladder and biliary tree. Report of a case. AB - Granular cell tumors are benign lesions that usually appear in the skin, oral activity, and mucous membrane. Occasionally, they will appear in visceral organs, and cause signs and symptoms similar to those of other organ-related diseases. We studied a patient with granular cell tumors of the common bile duct whose clinical appearance mimicked that of cholecystitis with cholelithiasis. Recognition of granular cell tumors on frozen section will allow surgical resection with probable cure. PMID- 6287962 TI - Asymmetric hamstring reflexes indicative of L5 radicular lesions. AB - There is no commonly accepted reflex from L5 innervated muscles. Hamstring muscles are primarily innervated by the L5 and S1 roots. It is shown that in the presence of symmetrically active gastrocsoleus reflexes, asymmetry of the hamstring reflexes indicates an L5 root lesion. We describe a method for eliciting the medial hamstring reflex by percussing the conjoint tendon of the semitendinosus and biceps femoris muscles at the level of the ischial tuberosity. Electromyographic, myelographic, and operative data confirmed the presence of an L5 root lesion in 7 patients with asymmetric hamstring reflexes. PMID- 6287963 TI - Crystal-induced arthritis. AB - The identification of monosodium urate crystals in joint effusions of patients with gouty arthritis established that crystals can cause arthritis. Other crystals causing arthritis have also been identified, including calcium, pyrophosphate dihydrate (chondrocalcinosis, pseudo-gout), calcium hydroxapatite crystals (calcific periarthritis, acute arthritis) and depot corticosteroid crystals (which occasionally cause arthritis when injected intra-articularly.) Crystal-induced arthritis is characterized by acute articular inflammation although rarely causing joint destruction or permanent disability. It is important for clinicians because it can mimic more serious joint diseases like septic arthritis or even rheumatoid arthritis. It can be diagnosed with precision and in some types as in gout can be treated effectively. Also, it constitutes one of the best understood articular inflammatory processes and often is the first clinical clue for the presence of curable metabolic or endocrine diseases. PMID- 6287964 TI - Estimation of cytomegalovirus--imposed risks on the outcome of renal transplantation. PMID- 6287965 TI - Ultrastructure of Entamoeba histolytica. A 96 hours old culture of an autochthonous strain. PMID- 6287966 TI - Major hepatic resections for neoplasia in children. AB - In a consecutive series from 1968 to 1978, 11 hepatic resections encompassing at least two hepatic segments were carried out for neoplasia in pediatric patients varying in age from 7 days to 14 years. There were no operative deaths. These resections consisted of four right lobectomies, three extended right lobectomies, one right lobectomy with right nephrectomy and inferior vena caval resection, two left lobectomies, and one left extended lobectomy. Diagnoses were hepatoblastoma in six patients, Wilms' tumor invading the liver in two, hemangioendothelioma in two, and malignant mesenchymoma in one. Selective angiography and technetium Tc 99m sulfur colloid scintigraphy were important preoperative aids. Complications were infrequent and there were no major infections or biliary fistulas. There was one death eight months postoperatively due to recurrent hepatoblastoma. Vigorous hepatic regeneration occurred in all instances. Major hepatic resections are well tolerated in children and allow good subsequent development. PMID- 6287967 TI - Near-total pancreatectomy in persistent infantile hypoglycemia. AB - To prevent the devastating effects of hypoglycemia on the CNS of the infant, persistent infantile hypoglycemia should be recognized early and treated promptly. To avoid mental retardation, surgical intervention should not be considered as a last resort. When an insulinoma is identified at surgery, subtotal pancreatectomy is adequate in achieving normoglycemia. However, the most common cause of insulin excess in infancy appears to be islet cell hyperplasia or nesidioblastosis, both diffuse processes. In these patients, subtotal pancreatectomy is often ineffective in achieving normoglycemia. Near-total pancreatectomy, retaining the spleen and duodenum, is a safe procedure, well tolerated by infants and children, and should be considered early for the correction of hypoglycemia of infancy that is not readily controlled by medical intervention. PMID- 6287968 TI - Radionuclide angiography for the diagnosis of thyroid cancer. AB - The conventional static thyroid scan is sensitive but nonspecific in detecting thyroid tumors. Radionuclide angiography is a noninvasive and simple technique that can improve the specificity of the conventional scan significantly, since it can demonstrate hypervascularity of tumors. Our case reports demonstrated hypervascularity of the primary tumor and metastatic cervical lymph nodes. PMID- 6287969 TI - Cytological study on the anterior pituitary of senile untreated beagle bitches with spontaneous mammary tumours. AB - Pituitaries were obtained from senile untreated Beagle bitches of comparable age (7-9 years) and genital status. The animals were divided into three groups; one was normal (without mammary lesions), one had benign tumours and one had mammary adenocarcinomas. PRL-, STH-, ACTH- and gonadotrophin-producing cells are studied and counted in serial paraffin sections stained with histochemical techniques. The animals with mammary malignancy displayed a marked increase in the relative number of PRL and ACTH cells with morphological signs of higher secretory activity in most cells, compared with that in normal bitches or bitches with benign tumours. STH cells in bitches with adenocarcinomas were reduced in number; however the secretory activity in these animals was the same as that observed in the normal bitches. In the animals with benign mammary tumours, STH cells showed morphological indication of higher secretory activity than in the other groups. PRL and ACTH cells were slightly increased in number and had slightly higher activity than that in normal bitches. These findings may suggest a role for hypophyseal hormones in mammary neoplasias. PMID- 6287970 TI - The effects of relative humidity and temperature on the survival of human rotavirus in faeces. AB - The effects of relative humidity and temperature on the survival of human rotavirus in a thin layer of faeces on an impervious surface and on absorbent material was investigated using the indirect immunofluorescence technique in LLC MK2 cells to titrate infectivity. Rotavirus was found to be very stable at low and high relative humidities but not in the medium range of relative humidity. Rotavirus infectivity was lost more rapidly under all humidities at 37 degrees C than at 4 degrees or 20 degrees C. PMID- 6287971 TI - Circular forms of viral DNA in Marek's disease virus-transformed lymphoblastoid cells. AB - The state of viral DNA in three Marek's disease virus (MDV)-transformed chicken cell lines (MDCC-MSB-1, MDCC-HP-2, MDCC-RP-1) was investigated by CsCl-density gradient centrifugation, and sedimentation analysis in neutral glycerol gradients. Each cell line contained MDV DNA integrated into the host cell genome. Additionally, free viral DNA could be identified in MDCC-MSB-1 and MDCC-HP-2 cells, sedimenting at about 100 S, and banding at the position of circular DNA in CsCl-ethidiumbromide gradients. Thus, MDV DNA with properties of circular plasmid DNA could be demonstrated in 2 virus-transformed cell lines. The significance of circular plasmid forms of viral DNA remains yet to be clarified, since these are apparently not regularly found in MDV-transformed cells. PMID- 6287972 TI - The small rna-viruses of insects. PMID- 6287973 TI - A microtiter test for detecting and titrating noncytopathogenic bovine viral diarrhea virus. AB - Bovine cells free of noncytopathogenic bovine viral diarrhea virus (NC-BVDV) treated with polyriboinosinic acid : polyribocytidylic acid (poly I:C) were protected against challenge with vesicular stomatitis virus (VSV), whereas NC BVDV-infected cells treated with poly I:C were not protected against VSV. An assay based on the ability of NC-BVDV to inhibit poly I:C protection of cells against VSV was developed and is herein referred to as PINBA (poly I:C for NC BVDV assay). Noncytopathogenic BVDV was titrated as cytopathogenic strains except that several days after infection with NC-BVDV, the cultures were treated with poly I:C and VSV. Titration endpoints were reached 24 hours later. PINBA was standardized for amount of VSV, time of addition of poly I:C, and time NC-BVDV had to be present to obtain stable titration endpoints. PINBA also was useful for titrating virus neutralizing antibodies. Compared with the fluorescent antibody test, PINBA was less subjective for detection of NC-BVDV. Compared with the interference test in which NC-BVDV infected cultures are challenged with a cytopathogenic strain of BVDV, PINBA was more reliable. The technique described herein is a simple and practical microtiter method for titrating NC-BVDV and virus neutralizing antibodies and for the presumptive detection of NC-BVDV. PMID- 6287974 TI - The fibers of fowl adenoviruses. AB - Purified virions from 14 strains of fowl adenoviruses (FAV) representing 11 serotypes were examined by electron microscopy. Pentons of all strains turned out to have two projections at their penton bases. It is shown that the double fibers also protrude from the intact virion. The length of both fibers was measured on free pentons and is given for each serotype. Double fibers seem to be a common feature of FAV. The serologically unrelated Egg Drop Syndrome (EDS) avian adenovirus strain 127 was confirmed to have single fibers only. PMID- 6287975 TI - [A study on the control of blood levels of histamine by intravenous administration of steroids and synthetic ACTH in patients with bronchial asthma (author's transl)]. PMID- 6287976 TI - [Importance of the PAS method in the histochemical analysis of carbohydrates and carbohydrate-containing biopolymers]. PMID- 6287977 TI - [Functional morphology of pituitary adenoma in acromegaly]. AB - Twenty-one pituitary adenomas removed from patients with acromegaly were studied electron microscopically. In 9 cases the tumors underwent immunohistochemical analysis by Coons' indirect immunofluorescent technique. Before the operation, all the patients were found by radioimmunoassay to have concentrations of pituitary hormones in the blood serum. A clinico-morphological analysis performed showed that in all cases of acromegaly pituitary adenoma consisted of somatotropocytes differing by the degree of cell granulation. The concentration of the somatotropic hormone (STH) in the blood serum of the patients did not depend on the intensity of the ultrastructural signs of the secretory activity of the cells. In 4 adenomas, a positive immunohistochemical reaction to two hormones, STH and prolactine, was demonstrated. PMID- 6287978 TI - [Cardiopulmonary resuscitation]. PMID- 6287979 TI - Balloon embolization of the sphenopalatine artery in a case of a juvenile angiofibroma. Clinical aspects. PMID- 6287980 TI - Critical remarks on the role of collagenolytic enzymes in carcinomas of the head and neck. AB - Collagenolytic activity of eight squamous cell carcinomas of the head and neck as well as xenografts of such tumors in nude mice was investigated by two different methods. High enzyme activities seem to be present in necrotic tumor areas, whereas the germinative zones tend to show lower collagenase levels. There is experimental evidence for the assumption of a collagen-stabilizing factor (CSF). A hypothetic concept including the results of other authors is developed concerning the role of collagenase in infiltrative growth of malignant tumors. The greater part of these questions being unanswered, further studies on collagenolytic activity in malignancy will be necessary. PMID- 6287981 TI - [The effect of alpha receptor-blocking agents on inner ear function (author's transl)]. AB - Nicergolin is used in the treatment of tinnitus, cerebral insufficiency, and sensorineural hearing loss. An increase of cerebral blood flow was observed after i.v. injection of Nicergolin (0.1 mg/kg). The effect of this drug on the inner ear was studied by measuring the DC potential and K+ activity in the endolymph. A rapid fall of the DC potential and hypocapnea were observed after the injection of Nicergolin (0.2 mg/kg). An increase of expiratory PCO2 (10-20 mm Hg) was also observed. Arteficial respiration with pure oxygen prevented the fall of the DC potential. The present investigation demonstrates that this drug primarily acts on the respiratory center with secondary PCO2 increase in the blood which then causes the dilatation of cerebral vessels. The fall of the DC potential after the injection of Nicergolin appears to be caused by hypoxia and not by direct effect on the inner ear. PMID- 6287984 TI - Low density lipoprotein metabolism in cultured fibroblasts from a new group of patients presenting clinically with homozygous familial hypercholesterolemia. AB - The metabolism of low density lipoproteins (LDL) was studied in cultured fibroblasts obtained from five local patients diagnosed, on the basis of clinical features and serum cholesterol concentrations, as having the homozygous form of familial hypercholesterolemia. LDL receptor function was assessed by measuring the binding, internalization, and degradation of 125I-labeled LDL, and by measuring the stimulation of cellular acyl-CoA cholesterol acyltransferase (ACAT) activity which followed exposure to LDL. Fibroblasts from two cases (CF and GM) showed receptor activities which were approximately 10% of the values obtained with normal cells, while ACAT stimulation by LDL was very low. These two patients were classified as homozygous for a receptor-defective abnormality. However, fibroblasts from the other three patients (JG, ES, and TT) showed greater than 25% of normal receptor activity, as assessed by 125I-LDL binding and catabolism. ACAT stimulation by LDL in cells from JG and ES was within the range of values shown by cells previously characterized as heterozygous for a receptor-negative mutation. Cells from ES behaved atypically: a low, but nonsaturable, activation by LDL was evident. ACAT stimulation by LDL was normal in cells from TT. Receptor activities of the cells from the available parents, assessed on the basis of LDL binding and degradation or of ACAT stimulation, were not clearly distinguishable from those of normal cells. These results add to the growing evidence of genetic heterogeneity underlying the clinical picture associated with familial hypercholesterolemia in different geographical distributions. PMID- 6287982 TI - George Lyman Duff Memorial Lecture. Atherosclerosis: a problem of the biology of arterial wall cells and their interactions with blood components. PMID- 6287985 TI - Successful weight reduction programmes. PMID- 6287983 TI - Cannabinoids impair the formation of cholesteryl ester in cultured human cells. AB - The ability of cultured human fibroblasts to form cholesteryl esters from 14C oleate is impaired by delta'-tetrahydrocannabinol, cannabidiol, and cannabinol, a group of natural products isolated from Cannabis sativa. This inhibition is compound and dose-related; 30 microM cannabidiol reduced esterification to less than 20% of the control values. The esterification of endogenous and exogenous cholesterol was affected, since inhibition was seen with either low density lipoproteins (200 micrograms/ml) or 25-hydroxycholesterol (5 micrograms/ml) as esterification stimuli. Cells treated with these compounds at doses of from 1 to 30 microM showed no impairment of protein synthesis, triglyceride or phospholipid formation, or ability to metabolize 125I-low density lipoproteins. An inhibition of cholesterol esterification was seen in human aortic medial cells. With increasing doses of these compounds, low density lipoproteins (25 micrograms/ml) became progressively less effective in suppressing HMG-CoA reductase in cultured human fibroblasts; with 30 microM cannabidiol the enzyme suppression was only 24% of that found in cells incubated with low density lipoproteins in the absence of drugs. Based on these data, we conclude that the cannabinoids "compartmentalize" cholesterol and, thus, make is unavailable for regulating cellular cholesterol metabolism. This may occur as a result of enhanced sterol efflux. PMID- 6287986 TI - Fibre in the prevention and management of chronic disease. AB - Some of the evidence of the role of dietary fibre in the prevention and treatment of disease is reviewed and the heterogeneous nature of fibre is considered. The relationships between various fibre fractions and colonic disorders, including cancer, diabetes mellitus, hypercholesterolaemia and obesity are discussed. It is concluded that an increase of dietary fibre constituents is only one part of a programme of dietary modification relevant to chronic diseases of a Western industrialised society. PMID- 6287987 TI - Metabolism of Babesia parasites in vitro. Comparison of several zwitterion buffers for the in vitro incubation of Babesia rodhaini. AB - The effect on the haemoprotozoan parasite Babesia rodhaini of adding a zwitterion buffer, HEPES, to a basal medium was studied by comparing the glucose uptake, ATP concentration, medium pH and infectivity of parasitized rat erythrocytes every 4 h during 24 h incubation in either Krebs Ringer phosphate (KRP) or KRP plus 28 mM HEPES. KRP plus HEPES was found to be superior to KRP for the maintenance of glucose uptake, erythrocyte ATP concentration, medium pH and infectivity of the parasite. Having established that HEPES improved the basal KRP medium, HEPES was then compared to three other buffers TES, BES and TRIS, to determine their relative effects on B. rodhaini during 18 h of incubation. At 28 and 56 mM concentration there was no significant difference in glucose uptake or lactate production among the 4 buffers. However, TES maintained the medium pH best, and BES preserved the infectivity of the parasite best. Further comparison of TES and BES at 10, 20, 30 and 40 mM established 50 mM TES to be best for buffering the pH of the medium and maintaining infectivity of B. rodhaini. PMID- 6287988 TI - Conventional treatment and its effect on survival of malignant pleural mesothelioma in Western Australia. AB - Analysis of the survival of all 81 cases of pathologically confirmed malignant pleural mesothelioma in Western Australia between January 1957 and December 1980 has revealed a median survival from diagnosis of 5.1 months (mean 7.8 months). The average time between presentation and diagnosis was 3.4 months. Survival was better in younger subjects and subjects selected for surgery but was unrelated to sex, symptoms at onset, history of asbestos exposure, tumour morphology, therapy other than surgery or year of presentation. The selection of subjects at earlier clinical stages for surgical intervention is considered to account for their longer survival. The outlook for patients with this disease remains poor and there is still no optimism for future advances in therapy. PMID- 6287989 TI - Acute sporadic non-A and non-B hepatitis in an urban community in New Zealand. AB - Acute sporadic non-A, non-B hepatitis is reported for the first time in New Zealand. Examination of sera from 94 patients with biochemical evidence of hepatitis showed that 26 (27%) had acute hepatitis B (HB), 22 (23%) had acute hepatitis A (HA) and 25 (26%) had acute non-A, non-B hepatitis (NANBH). Nine (10%) patients had Epstein-Barr virus (EBV) associated hepatitis and one (1%) had cytomegalovirus (CMV) hepatitis. There were 11 (13%) patients with mixed infections; eight with HA plus HB, one with HB plus EBV, one with HA plus EBV and one with HB plus CMV. Thus NANBH and EBV associated hepatitis must be considered in the differential diagnosis of patients presenting with clinical hepatitis with no history of possible percutaneous infection. PMID- 6287990 TI - Change in plasma cAMP and catecholamines in men subjected to the same relative amount of physical work stress. AB - This study has shown, for the first time, that the change in plasma adenosine 3',5' cyclic monophosphate (cAMP), from accommodation or trough levels, correlates highly in both progressive and continuous work stress experiments with epinephrine (E) at 0.990 and 0.898, and norepinephrine (NE) at 0.954 and 0.927, respectively, in men subjected to the same relative, rather than absolute, stress levels. All three analytes increased significantly (p less than 0.05) by the time each subject had reached 80% of their Vo2 max in the progressive experiments and after working at 80% of their Vo2 max continuously for 10 min. In no case did plasma cAMP rise before the catecholamines. The possible significance of these observations to net perceived relative physiological stress are discussed. PMID- 6287992 TI - Low level and high level DNA rearrangements in Escherichia coli. AB - It can be argued that all organisms exhibit two levels of DNA rearrangements. At a low level they may occur sporadically in cells, perhaps largely because of spontaneous activity of transposable genetic elements. A high level may be induced in special circumstances if functions that cause rearrangements are hyperactive. As an example of low level genetic rearrangements, we have studied the occurrence of spontaneous polar mutations in the early regions of prophage Mu. We isolated 49 independent prophage mutants, which are defective in replication ad expression of late genes; 44 were in the B region and 5 were in the A region. In the B region, 68% were IS1 insertions, 9% were IS5 insertions and 9% were IS2 insertions; 14% showed no insertion. In the A region, all 5 were IS5 insertions. Thus most spontaneous polar mutations in Escherichia coli appear to the insertions. IS1 is the most common insertion; however, certain DNA rearrangements are exemplified by DNA fusion and DNA dissociation that occur when replication-transposition functions of Mu are induced. PMID- 6287991 TI - Single-stranded gaps as localized targets for in vitro mutagenesis. AB - Short single-stranded gaps in circular DNA molecules can be generated enzymatically, often at predetermined sites. These can serve as targets for in vitro mutagenesis procedures that result in alterations in nucleotide sequence within or very near the gap. Deamination of unpaired cytosine residues with sodium bisulfite has been used to induce mutations in the BglI restriction site of SV40 DNA and within defined regions of the beta-lactamase gene on pBR322. A new method of induction of mutations at gaps, called "gap misrepair," has been developed; it was used to cause changes at the HindIII and C1aI restriction sites on pBR322 DNA. Gap misrepair reactions using DNA polymerase I of Micrococcus luteus in the presence of T4 DNA ligase and three of the four deoxynucleoside triphosphates yielded all three possible substitutions for adenine and cytosine residues in the DNA. PMID- 6287993 TI - Mutants of Escherichia coli K12 which affect excision of transposon Tn10. AB - We have described three illegitimate recombination events associated with, but not promoted by, transposon Tn10: precise excision, nearly precise excision, and precise excision of a nearly precise excision remnant. All three are structurally analogous: excision occurs between two short direct repeat sequences, removing all intervening material plus one copy of the direct repeat. In each case, the direct repeats border a larger inverted repeat. We report here the isolation of host mutants of Escherichia coli K12 which exhibit increased frequencies of precise excision of Tn10. Nineteen of the 39 mutants have been mapped to five distinct loci on the E. coli genetic map and have been designated texA through texE (for Tn10 excision). Mapping and genetic characterization indicate that each tex gene corresponds to a previously identified gene involved in cellular DNA metabolism: recB and/or recC, uvrD, mutH, mutS, and dam. The role of these various DNA repair and recombination genes in an illegitimate recombination process such as Tn10 excision will be discussed. In addition to an increase in precise excision frequency, all 39 tex mutants display an increased frequency for nearly precise excision. However, none of the mutants are increased for the third excision event, precise excision of a nearly precise excision remnant, supporting the idea that precise and nearly precise excision occur by closely related pathways which are distinct from those pathways which promote the third type of excision event. PMID- 6287994 TI - Gene conversion: a possible mechanism for eliminating selfish DNA. PMID- 6287995 TI - Transposons and illegitimate recombination in prokaryotes: a summary and perspective. PMID- 6287996 TI - Perspectives in molecular mutagenesis. AB - The models and paradigms that underlie a vigorously developing science may tend to stifle progress or may serve to sharpen the knife edge of paradox. Working out mutagenic mechanisms is a conceptually and technologically demanding task, and we are accumulating an increasingly uncomfortable number of experimental and theoretical inconsistencies. First, there continue to be widespread difficulties in specifying the chemical nature of mutagenic DNA alterations, both because of the multitude of DNA reaction products induced by many mutagens and because of the intrinsic rarity of most mutational responses. For instance, alkylation of the 0(6) position of guanine to generate adducts of modest dimensions is widely believed to form the basis for the mutagenic and carcinogenic actions of numerous chemicals. However, while this scheme is supported by in vitro evidence, it has failed to explain why bacteriophages can be thus alkylated in vitro by N-methyl N'-nitro-N-nitrosoguanidine without the production of mutations, or why microbial eukaryotes alkylated by ethyl methanesulfonate or N-methyl-N'-nitro-N nitrosoguanidine display no mutagenic response when their "error-prone repair systems" are mutationally inactivated. Second, a base pair is typically mutated at vastly different rates, and with different directional specificities, when it resides at different positions within a gene; whereas very little of this variability is explained by current theories that aim to describe the determinants of fidelity in DNA replication. (Some sizable portion of this variation now appears to depend not only upon neighboring base-pair influences but also upon much more subtle and distant effects). Third, experimental modifications of enzymatic fidelity by means of amino acid substitutions, and perhaps also cation replacements, lead to such a diversity of modified mutation rates as to seriously challenge the ability of any simple theory to organize the experimental observations into a coherent and predictive network. PMID- 6287997 TI - Regulation and functions of Escherichia coli genes induced by DNA damage. PMID- 6287998 TI - [Immunization of mice against sarcosporidia (Sarcocystis muris) with attenuated sporocysts]. PMID- 6288000 TI - The kinetics of electron transfer between pseudomonas aeruginosa cytochrome c-551 and its oxidase. AB - The redox reaction between cytochrome c-551 and its oxidase from the respiratory chain of pseudomonas aeruginosa was studied by rapid-mixing techniques at both pH7 and 9.1. The electron transfer in the direction of cytochrome c-551 reduction, starting with the oxidase in the reduced and CO-bound form, is monophasic, and the governing bimolecular rate constants are 1.3(+/- 0.2) x 10(7) M-1 . s-1 at pH 9.1 and 4 (+/- 1) x 10(6) M-1 . s-1 at pH 7.0. In the opposite direction, i.e. mixing the oxidized oxidase with the reduced cytochrome c-551 in the absence of O2, both a lower absorbance change and a more complex kinetic pattern were observed. With oxidized azurin instead of oxidized cytochrome c-551 the oxidation of the c haem in the CO-bound oxidase is also monophasic, and the second-order rate constant is 2 (+/- 0.7) x 10(6) M-1 . s-1 at pH 9.1. The redox potential of the c haem in the oxidase, as obtained from kinetic titrations of the completely oxidized enzyme with reduced azurin as the variable substrate, is 288 mV at pH 7.0 and 255 mV at pH 9.1. This is in contrast with the very high affinity observed in similar titrations performed with both oxidized azurin and oxidized cytochrome c-551 starting from the CO derivative of the reduced oxidase. It is concluded that: (i) azurin and cytochrome c-551 are not equally efficient in vitro as reducing substrates of the oxidase in the respiratory chain of Pseudomonas aeruginosa; (ii) CO ligation to the d1 haem in the oxidase induces a large decrease (at least 80 mV) in the redox potential of the c-haem moiety. PMID- 6287999 TI - Location and properties of two isoenzymes of cardiac adenylate kinase. AB - Adenylate kinase catalyses the equilibrium 2ADP = ATP + AMP. There are two isoenzymes of adenylate kinase in bovine ventricular tissue, one cytoplasmic, the other mitochondrial. Mitochondrial subfractionation locates this isoenzyme between the mitochondrial membranes with fatty acid-CoA ligase. The cytoplasmic and mitochondrial isoenzymes are distributed in ratio 3:2, and both forms were purified to homogeneity. They differ principally by charge, Km values for ATP, ADP and AMP, pH-stability and -activity profiles, and susceptibility to the inhibitor adenosine pentaphosphoadenosine. The forward and reverse reactions show similar energies of activation for the cytoplasmic enzyme, but differ for the mitochondrial enzyme. The molecular weights are indistinguishable. An integrated mechanism is formulated whereby one isoenzyme suppresses the activation of fatty acid and the other enhances carbohydrate utilization in hypoxic myocytes. PMID- 6288001 TI - A stereochemical investigation of the hydrolysis of cyclic AMP and the (Sp)-and (Rp)-diastereoisomers of adenosine cyclic 3':5'-phosphorothioate by bovine heart and baker's-yeast cyclic AMP phosphodiesterases. AB - Bovine heart cyclic AMP phosphodiesterase, which has a requirement for Mg2+, hydrolyses cyclic AMP with inversion of configuration at the phosphorus atom, but only the (Sp)-diastereoisomer of adenosine cyclic 3':5'-phosphorothioate is hydrolysed by this enzyme. By contrast, the low-affinity yeast cyclic AMP phosphodiesterase, which contains tightly bound Zn2+, hydrolyses both the (Sp)- and the (Rp)-diastereoisomers of adenosine cyclic 3':5'-phosphorothioate, the (Rp)-diastereoisomer being the preferred substrate under V max. conditions. Both of the diastereoisomers of adenosine cyclic 3':5'-phosphorothioate, as well as cyclic AMP, are hydrolysed with inversion of configuration at the phosphorus atom by the yeast enzyme. It is proposed that, with both enzymes, the bivalent metal ion co-ordinates with the phosphate residue of the substrate, and that hydrolysis is catalysed by a direct "in-line' mechanism. PMID- 6288002 TI - An investigation by e.p.r. and optical spectroscopy of cytochrome oxidase during turnover. AB - Cytochrome oxidase (EC 1.9.3.1; ferrocytochrome c:oxygen oxidoreductase) was studied during steady-state by optical and e.p.r. methods. Starting with either the 'resting' or the 'pulsed' enzyme, oxidase, cytochrome c, ascorbate and O2 were mixed and the reaction monitored optically. Tetramethylphenylenediamine was used as mediator to poise the steady-state to the desired reduction level. After mixing, the reaction was quenched by the used of rapid-freeze techniques. The e.p.r. spectra of samples captured at increasing tetramethylphenylenediamine concentrations (i.e. higher electron flux) show decreasing g = 2 (Cu A) and g = 3 (cytochrome a) signals. No Cu B or g = 6 signals (high-spin cytochrome a3) could be found during the reaction. Also, the signal with peaks at g = 1.69, 1.78 and 5 as well as the g = 12 signal was hardly detectable at higher turnover rates. The only new signal appearing during turnover is a radical signal, which is discussed in terms of a protein radical. Finally, a scheme is presented, proposing a catalytic cycle for cytochrome oxidase with respect to the O2 binding Cu B cytochrome a3 unit. PMID- 6288003 TI - The steady state kinetics of the NADH-dependent nitrite reductase from Escherichia coli K12. The reduction of single-electron acceptors. AB - The kinetic characteristics of the diaphorase activities associated with the NADH dependent nitrite reductase (EC 1.6.6.4) from Escherichia coli have been determined. The values of the apparent maximum velocity are similar for the reduction of Fe(CN)6(3)-and mammalian cytochrome c by NADH. These reactions may therefore have the same rate-limiting step. NAD+ activates NADH-dependent reduction of cytochrome c, and the apparent maximum velocity for this substrate increases more sharply with the concentration of NAD+ than for hydroxylamine. The simplest explanation is that NAD+ activation of hydroxylamine reduction derives solely from activation of steps involved in the reduction of cytochrome c, a flavin-mediated reaction, but these steps are only partly rate-limiting for the reduction of hydroxylamine. At 0.5 mM-NAD+, the apparent maximum velocity was 2.3 times higher for 0.1 mM-cytochrome c as substrate than for 100 mM-hydroxylamine, suggesting that the rate-limiting step during hydroxylamine reduction is a step that is not involved in cytochrome c reduction. A scheme is proposed that can account for the pattern of variation with [NAD+] of the Michaelis-Menten parameters for hydroxylamine and for NADH with hydroxylamine or cytochrome c as oxidized substrate. PMID- 6288004 TI - The inhibitory effect of Zn2+ on poly(ADP-ribose) polymerase activity and its reversal. AB - Zn2+ inhibits purified poly(ADP-ribose) polymerase (50% inhibition at 10 microM). Furthermore poly (ADP-ribose) polymerase present in nuclei and metaphase chromosome clusters is also inhibited by Zn2+. The inactivated enzyme could be re activated by dithiothreitol. The concentration of Zn2+ needed to affect the enzyme activity in the organelles is sufficiently low for it to have a possible role in controlling the activity of this chromatin-bound enzyme. PMID- 6288005 TI - Titration and steady-state behaviour of the 830 nm chromophore in cytochrome c oxidase. AB - Titration of cyanide-incubated cytochrome c oxidase (ox heart cytochrome aa3) with ferrocytochrome c or with NNN'N'-tetramethyl-p-phenylenediamine initially introduces two reducing equivalents per mol of cytochrome aa3. The first equivalent reduces the cytochrome a haem iron; the second reducing equivalent is not associated with reduction of the 830 nm chromophores (e.p.r.-detectable copper) but is probably required for reduction of the e.p.r.-undetectable copper. Excess reductant introduces a third reducing equivalent into the cyanide complex of cytochrome aa3. During steady-state respiration in the presence of cytochrome c and ascorbate, the 830 nm chromophore is almost completely oxidized. It is reduced more slowly than cytochrome a on anaerobiosis. In the presence of formate or azide, some reduction at 830 nm can be seen in the steady state; in an oxygen pulsed system, a decrease in steady-state reduction of cytochromes c and a is associated with ab increased reduction of the 830 nm species. In the formate inhibited system the reduction of a3 on anaerobiosis shows a lag phase, the duration of which corresponds to the time taken for the 830 nm species to be reduced. It is concluded that the e.p.r.-undetectable copper (CuD) is reduced early in the reaction sequence, whereas the detectable copper (CUD) is reduced late. The latter species is probably that responsible for reduction of the cytochrome a3 haem. The magnetic association between undetectable copper and the a3 haem may not imply capability for electron transfer, which occurs more readily between cytochrome a3 and the 830 nm species. PMID- 6288007 TI - Cyclic nucleotide phosphodiesterase activities in Neurospora crassa. AB - Cyclic nucleotide phosphodiesterase activities in soluble Neurospora crassa mycelial extracts were resolved into two peaks, phosphodiesterase I and II, by chromatography on DEAE-cellulose columns. Phosphodiesterase I hydrolysed cyclic AMP and cyclic GMP equally well. Phosphodiesterase II was active on cyclic GMP but scarcely active on cyclic AMP. Phosphodiesterase I was resolved by gel filtration and sucrose-density-gradient centrifugation into three peaks having molecular weights of about 57 000, 125 000 and 225 000. This suggests that this enzyme activity has at least three aggregation forms, tentatively defined as monomeric, dimeric and tetrameric. Similarly, phosphodiesterase II was resolved into two forms, having molecular weights of about 170 000 and 320 000. Evidence on the interconversion between phosphodiesterase I forms was obtained. PMID- 6288006 TI - A quantitative test for superoxide radicals produced in biological systems. AB - The preparation and properties of a partially succinoylated cytochrome c, suited for the detection of superoxide anion radicals in liver microsomes, is reported. By succinoylation of 45% of the primary amino groups of horse heart cytochrome c the activity towards solubilized NADPH--cytochrome P-450 reductase was diminished by 99% compared with native cytochrome c. The capacities of cytochrome b5 and cytochrome c oxidase to reduce the succinoylated ferricytochrome c and oxidize succinoylated ferrocytochrome c respectively were decreased to a similar extent. However, the bimolecular rate constant for the reduction of the partially succinoylated ferricytochrome c by O2-. was estimated to be one-tenth of the value for the reaction of O2-. with native ferricytochrome c a pH 7.7. On this basis the quantification of O2-. generated by NADPH-supplemented liver microsomes became possible. The initial rates of succinoylated ferricytochrome c reduction determined at various finite concentrations of the cytochrome c derivative can be extrapolated to obtain true rates of O2-. generation in a homogeneous system. The problems encountered in the quantitative determination of O2-. produced in biological membranes, e.g. microsomes, are discussed. PMID- 6288008 TI - Different characteristics of solubilized lactogen receptors from livers of pregnant and non-pregnant female rats. AB - Receptors specific for lactogenic hormones were solubilized by 1% (v/v) Triton X 100 from the crude particulate membrane fraction of livers of pregnant and non pregnant female rats and the characteristics of both preparations were compared. Human (125)I-labelled somatotropin was used for binding studies of lactogenic hormone. The solubilized receptor retained most of the characteristics noted in the particulate fraction. The binding of human (125)I-labelled somatotropin to the solubilized receptor is a saturable process, depending on temperature and time. Scatchard analysis of displacement curves revealed similar affinity constants ranging from 1.02 x 10(9) to 1.20 x 10(9) 1/mol, while the binding capacity was 4.5 times greater in the pregnant rat livers than in the non pregnant female rat livers. The receptors for human (125)I-labelled somatotropin from livers of non-pregnant and pregnant female rats were equally adsorbed onto a concanavalin-A-Sepharose column and were dissociated from the column with alpha methyl-d-glucoside or alpha-methyl-d-mannoside in the same manner. By gel filtration on Sepharose 6B, however, the molecular sizes of the hepatic receptors were found to be different. The apparent M(r) value was approx. 270000 with a Stokes' radius of 5.5nm in the non-pregnant female rats and approx. 330000 with a Stokes' radius of 5.5nm in the pregnant rats. Furthermore, isoelectric-focusing experiments showed that a major part of the receptor from the non-pregnant female rat livers had a neutral pI (7.0-8.5), whereas that from pregnant-rat livers had an acidic pI (4.2-4.7). These data suggest that the increase in the lactogenic binding capacity in rat liver membranes during pregnancy may be associated with marked changes of the physicochemical properties of the receptors. PMID- 6288010 TI - Forskolin as an activator of cyclic AMP accumulation and secretion in blowfly salivary glands. AB - Forskolin is a diterpene that activates adenylate cyclase in a variety of mammalian cells. In addition of forskolin to blowfly salivary glands increased cyclic AMP accumulation and salivary secretion. There was a small increase in transepithelial movement of labelled Ca2+. Forskolin did not induce breakdown of labelled phosphatidylinositol or inhibit the stimulation of phosphatidylinositol breakdown caused by 5-hydroxytryptamine. These data indicate that forskolin can mimic all the effects of 5-hydroxytryptamine on salivary-gland secretion that have been attributed to cyclic AMP. PMID- 6288009 TI - Studies on the interaction of staphylococcal delta-haemolysin with isolated islets of Langerhans. AB - delta-Haemolysin, a small surface-active polypeptide purified from the culture media of Staphylococcus aureus, was observed to stimulate the release of insulin from isolated rat islets of Langerhans. This effect was dose-dependent and saturable, with the half-maximal response elicited by a delta-haemolysin concentration of 10 micrograms/ml. Stimulation of insulin release by delta haemolysin (10 micrograms/ml) was not dependent on the presence of glucose in the incubation medium, but was augmented by increasing concentrations of the sugar. The release of insulin in response to delta-haemolysin could be inhibited by depletion of extracellular Ca2+ or by adrenaline (epinephrine) (10 microM) and was readily reversible when delta-haemolysin was removed from the medium. In addition, the response was potentiated by incubation with the phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine (0.2 mM). These observations suggest that delta-haemolysin induced a true activation of the beta-cell secretory mechanism. Stimulation of islets of Langerhans with delta-haemolysin was found to be associated with a modest increase in intracellular cyclic AMP levels, although the adenylate cyclase activity of islet homogenates was not increased by delta haemolysin. delta-Haemolysin was observed to induce a dose-dependent net accumulation of 45Ca2+ by islet cells and to stimulate the efflux of 45Ca2+ from preloaded islets. The efflux of 45Ca2+ was modest in size and short-lived, but dramatically increased in medium depleted fo 40Ca2+. Incubation in the presence of verapamil augmented delta-haemolysin-induced 45Ca2+ efflux and insulin secretion. delta-Haemolysin was found to be a potent 45Ca2+-translocating ionophore in an artificial system. This response was dose-dependent and could be augmented by verapamil. In addition, phosphatidylcholine (25 micrograms/ml) was found to inhibit both delta-haemolysin induced 45Ca2+ translocation and insulin release in a precisely parallel manner. These studies suggest that the ability of delta-haemolysin to stimulate insulin release may be due, in part, to the facilitation of Ca2+ entry into the beta-cells of islets of Langerhans, mediated directly by an ionophoretic mechanism. PMID- 6288012 TI - Discrimination in the metabolism of orally dosed ergocalciferol and cholecalciferol by the pig, rat and chick. AB - Vitamin D-deficient pigs, rats and chicks were repleted with four daily oral doses of crystalline ergocalciferol (vitamin D2) and cholecalciferol (vitamin D3) containing equal concentrations of each. At 24 h after the last dose, the plasma of each species was analysed for vitamin D and 25-hydroxyvitamin D by standard methods. The mean (+/- S.D.) ratios of plasma cholecalciferol to ergocalciferol concentration were 1.5 +/- 0.1 (pig). 1.7 +/- 0.1 (rat) and 6.3 +/- 1.2 (chick). The mean ratios of plasma 25-hydroxycholecalciferol to 25-hydroxyergocalciferol concentration were 4.0 +/- 0.1 (pig), 0.4 +/- 0.02 (rat) and 10.7 +/- 3.4 (chick). The mean plasma cholecalciferol/ergocalciferol ratios for the 24,25 dihydroxy-, 25,26-dihydroxy- and 1,25-dihydroxy-derivatives in the pig were 2.6 +/- 0.6, 5.8 +/- 1.3 and 5.8 +/- 0.8 respectively. This is the first evidence that mammals other than the New World monkey, like birds, discriminate between ergocalciferol and cholecalciferol. These data, therefore, suggest that species discrimination between the different forms of vitamin D is probably a general phenomenon in mammals. Moreover, this is the first indication of a species (rat) that discriminates against a cholecalciferol metabolite in favour of an ergocalciferol metabolite. Species discrimination against particular forms of vitamin D may be important to the choice of experimental animal models for studying the regulation of vitamin D metabolism and may also be an important consideration in dietary vitamin supplementation. PMID- 6288013 TI - Studies on the assembly of cytochrome oxidase in isolated rat hepatocytes. AB - 1. The assembly of rat liver cytochrome oxidase was studied in isolated hepatocytes and isolated liver mitochondria labelled with L-[35S]methionine. 2. Labelled subunits II and III appeared in the immunoabsorbed holoenzyme within minutes after the initiation of a pulse label. In contrast, labelled subunit I appeared in immunoabsorbed holoenzyme only after a subsequent 2 h chase or after an additional 2 h of labelling. Subunit I was heavily labelled, however, in intact mitochondria after 10 min. 3. A similar pattern of labelling was observed in holo-cytochrome oxidase which was chemically isolated by a small scale procedure adapted for this purpose. The appearance of subunit I in the holoenzyme was delayed for 1.5-2 h after a 60 min pulse with labelled methionine. 4. Incubation of hepatocytes for 4 h in the presence of cycloheximide had no effect on the labelling pattern described above. 5. Methods were developed in which newly translated, presumably unassembled, subunits of cytochrome oxidase could be separated from the holoenzyme by fractionation in Triton X-114. Short-term pulse experiments indicate that subunits II and III are associated with the holoenzyme fraction immediately after their completion, whereas subunit I is not. 6. The data are consistent with a model in which cytochrome oxidase assembly is viewed as an ordered and sequential event. PMID- 6288011 TI - Regulation of adenylate cyclase and cyclic AMP phosphodiesterase by 5 hydroxytryptamine and calcium ions in blowfly salivary-gland homogenates. AB - Salivary-gland homogenates contain 5-hydroxytryptamine-stimulated adenylate cyclase. Half-maximal stimulation was obtained with 0.1 microM-5 hydroxytryptamine in the presence of added guanine nucleotides. Gramine antagonized the stimulation of cyclase caused by 5-hydroxytryptamine. In the presence of hormone, guanosine 5'-[gamma-thio]triphosphate produced a marked activation of adenylate cyclase activity. Stimulation of adenylate cyclase by forskolin or fluoride did not require the addition of guanine nucleotides or hormone. In the presence of EGTA, Ca2+ produced a biphasic activation of cyclase activity. Ca2+ at 1-100 microM increased activity, whereas 2000 microM-Ca2+ inhibited cyclase activity. The neuroleptic drugs trifluoperazine and chlorpromazine non-specifically inhibited adenylate cyclase activity even in the absence of Ca2+. The cyclic AMP phosphodiesterase activity in homogenates was not affected by Ca2+ or exogenous calmodulin. This enzyme was also inhibited by trifluoperazine in the absence of Ca2+. These results indicate that Ca2+ elevates adenylate cyclase activity, but had no effect on cyclic AMP phosphodiesterase of salivary-gland homogenates. PMID- 6288014 TI - Modulation of collagen production by fibroblasts. Effects of chronic exposure to agonists that increase intracellular cyclic AMP. AB - Cultured human lung fibroblasts were evaluated for their responsiveness to isoprenaline (isoproterenol) or prostaglandin E2 before and after chronic incubation with the agonist. Cells incubated for 6 h with either agonist were suppressed in terms of collagen production and exhibited increased intracellular cyclic AMP. Cells incubated for 72 h with the agonist and then re-challenged for 6 h with the same agonist did not demonstrate suppressed collagen production or increased cyclic AMP. Cells incubated for 72 h with isoprenaline still responded to prostaglandin E2 when challenged for 6 h; however, when the order of agonist exposure was reversed, cells incubated with prostaglandin E2 did not respond to a challenge by isoprenaline. If cells were allowed to recover for 48 h without the agonist after a 72 h chronic incubation, they recovered their responsiveness to the agonist. The results indicate that, although cultured fibroblasts may become desensitized to one agonist, they may retain their sensitivity to a second agonist and chronic suppression of collagen production may be achieved by alternate exposure to isoprenaline and prostaglandin E2. PMID- 6288015 TI - The effects of calcium on protein turnover in skeletal muscles of the rat. AB - Several experimental procedures were used to increase the intracellular concentration of Ca2+ and determine its effects on protein turnover in isolated extensor digitorum longus and soleus muscle. These methods included the use of ionophore A23187, caffeine, dibucaine, thymol and procaine, all agents known to induce the release of calcium by acting either on the sarcolemma and/or on the sarcoplasmic reticulum. Another approach involved varying the external concentration of Ca2+ in the media in which the muscles were incubated. The changes in muscle Ca2+ concentrations after exposure to the various calcium releasing agents were in keeping with accepted modes of action of these agents on muscle membranes. The findings suggest that increasing the sarcoplasmic concentration of Ca2+ inhibits protein synthesis and enhances protein breakdown. These catabolic effects of Ca2+ are compared with the changes induced in muscle protein turnover after exposure to insulin or cyclic nucleotides, and in myopathic muscle and situations of work overload. Attention is also drawn to some of the difficulties involved in definitively implicating Ca2+ as a factor involved in the normal regulation of protein turnover. PMID- 6288016 TI - Human gamma-melanotropin precursor potentiates corticotropin-induced adrenal steroidogenesis by stimulating mRNA synthesis. AB - Circulating human gamma-melanotropin precursor (76 amino acids) [N-POC(1-76) enhances the corticosterone and aldosterone response to corticotropin by isolated perfused rat adrenal cells [Al-Dujaili, Hope, Estivariz, Lowry & Edwards (1981) Nature (London) 291, 156-159]. Actinomycin D (4 x 10(-6) mol/litre) did not significantly affect corticotropin-induced corticosterone or aldosterone outputs, but completely inhibited the potentiating action of gamma-melanotropin precursor on rat adrenal cells. This suggests that the precursor enhances corticotropin induced steroidogenesis via an increase in available mRNA produced from DNA transcription. PMID- 6288017 TI - Purification of rat kidney branched-chain oxo acid dehydrogenase complex with endogenous kinase activity. AB - A method was devised to purify branched-chain oxo acid dehydrogenase (BCOAD) from rat kidney which retains endogenous kinase activity. Incorporation of 32P into purified enzyme parallels the time course of enzyme inhibition by ATP. Phosphorylation occurs on a serine residue(s) of the 46000-mol.wt. subunit of the enzyme complex. Endogenous phosphatase activity is not present after purification, and added pyruvate dehydrogenase phosphate phosphatase does not re activate BCOAD or liberate 32P from previously labelled enzyme. These results demonstrate that BCOAD can be regulated by an endogenous protein kinase and that the phosphorylation-cycle enzymes regulating BCOAD appear to be distinct from those associated with pyruvate dehydrogenase complex. PMID- 6288018 TI - Phosphatidylinositol-hydrolysing enzymes in blowfly salivary glands. AB - The salivary glands of adult blowflies (Calliphora erythrocephala) contain enzymes that hydrolyse phosphatidylinositol, predominantly by a Ca2+-independent deacylation, though a Ca2+-dependent phosphodiesterase (phospholipase C) activity could be detected. The deacylating enzymes could also hydrolyse phosphatidylcholine and phosphatidylethanolamine, and were secreted in the saliva. Homogenization of salivary glands prelabelled with [3H]inositol resulted in a rapid deacylation of the endogenous 3H-labelled phosphatidylinositol; this hydrolysis was unaffected by addition of 5-hydroxytryptamine to the homogenate. PMID- 6288019 TI - The pathway of electron flow through ubiquinol:cytochrome c oxidoreductase in the respiratory chain. Evidence from inhibition studies for a modified 'Q cycle'. AB - 1. Cytochrome spectra of the liver and heart mitochondria incubated under various conditions are presented to compare the effects of antimycin, colletotrichin and 2-heptyl-4-hydroxyquinoline N-oxide (HQNO) additions. 2. Under aerobic conditions, in State 4, in the presence of uncoupler or in the presence of cyanide, all three inhibitors caused oxidation of cytochromes c and c1, but different changes in the spectra of the b cytochromes. Antimycin caused oxidation of a peak at 558 nm and reduction of peaks at 562 nm and 566 nm, whereas colletotrichin caused reduction of peaks at 558 nm and 566 nm and oxidation at 562 nm. HQNO had an effect on the spectra intermediate between those of the two other inhibitors. 3. Under aerobic conditions in the presence of 5 mM-succinate and 5 mM-fumarate, antimycin caused reduction of a peak at 566 nm and oxidation of a peak at 558 nm, whereas colletotrichin had the reverse effect and HQNO caused reduction of a peak at 562 nm. 4. Colletotrichin inhibition of the ADP stimulated oxidation of glutamate + malate was enhanced by succinate addition and declined again with rotenone addition. Similar but smaller effects were seen with inhibition by antimycin and HQNO. 5. Cytochrome spectra are shown of the effects of ADP and uncoupler addition to stimulate respiration progressively. 6. The results are interpreted in terms of a modified 'Q cycle' [Mitchell (1976) J. Theor. Biol. 62, 327-367] in which the three inhibitors are postulated to displace ubiquinone and ubisemiquinone specifically bound to cytochromes b on both sides of the membrane. 7. It is suggested that cytochromes b558 and b566 are the same b cytochrome located on the outer surface of the membrane, but binding ubisemiquinone or colletotrichin and ubiquinone or antimycin respectively. Cytochrome b562 is postulated to be on the inner surface of the mitochondrial membrane and to bind either ubiquinone or ubisemiquinone, HQNO would bind to the reduced form of the cytochrome and colletotrichin to the oxidized form. 8. Sites for the locus of action of glucagon and the protonmotive force on electron flow are suggested. PMID- 6288020 TI - The turnover of L-type pyruvate kinase in cultured rat hepatocytes. AB - Hepatocytes were isolated by collagenase perfusion of livers from rats that had been allowed access to a carbohydrate-rich diet or laboratory chow or had been deprived of food 48h before use. By incubation with l-[4,5-(3)H]leucine and precipitation with anti-(L-type pyruvate kinase) sera the rates of synthesis and degradation of L-type pyruvate kinase were measured in freshly prepared cells and hepatocytes maintained in monolayer culture for up to 5 days. Hepatocytes from carbohydrate-rich-diet-fed rats synthesized more L-type pyruvate kinase than did cells from chow-fed animals, which in turn synthesized more than cells from 48h starved rats. Hepatocytes maintained in culture for up to 5 days synthesized L type pyruvate kinase at similar rates to freshly prepared cells. The degradation of [(3)H]leucine-labelled L-type pyruvate kinase was shown to be biphasic. A phase with t((1/2)) (half-time) 4.9h and a duration of 8-10h was followed by a phase with t((1/2)) 79.2h. Cells from chow-fed and carbohydrate-rich-diet-fed rats showed similar patterns of degradation of L-type pyruvate kinase. The addition of 2mm-fructose and 0.1mum-insulin to the culture medium increased the t((1/2)) of the rapid phase to 12h in cells isolated from carbohydrate-rich-diet fed rats, but not in cells from chow-fed rats. The secondary, slower, phase of degradation remained unaffected. The degradation of fructose 1,6-bisphosphatase and total cell protein followed first-order kinetics. The half-life of fructose 1,6-bisphosphatase was 41.0h in cells from chow-fed animals and 48.5h in cells from carbohydrate-rich-diet-fed donors. Fructose and insulin did not affect the rate of enzyme degradation. We propose that there is a role for protein catabolism in the short-term and long-term control of L-type pyruvate kinase concentration. PMID- 6288022 TI - Factors determining the plasma-membrane potential of lymphocytes. AB - 1. Lymphocytes from pig mesenteric lymph node have low permeability to K+ (Rb+), Na+ and Cl-. None of these ions is in Nernst equilibrium with the plasma-membrane potential (delta psi p). 2. delta psi p can be calculated from the transmembrane distribution of the permeant cation methyltriphenylphosphonium (TPMP+) in the presence of the uncoupler carbonyl cyanide p-trifluoromethoxyphenylhydrazone (FCCP) to abolish uptake into intracellular mitochondria. In normal culture medium delta psi p is 56 mV. 3. A similar potential is found in T-enriched pig cells and in mouse thymocytes. 4. The contribution of electrogenic (Na+ + K+)ATPase to delta psi p is about 7 mV. 5. The remainder of the lymphocyte delta psi p is a polyionic potential set up by K+ and Cl- with a permeability coefficient for Cl- of similar magnitude to that for K+. PMID- 6288021 TI - THe proton-per-electron stoicheiometry of 'site 1' of oxidative phosphorylation at high protonmotive force is close to 1.5. AB - The maximum redox potential difference between the NAD+/NADH couple and the succinate/fumarate couple generated during ATP-energized reduction of NAD+ by succinate in submitochondrial particles was measured, together with the electrochemical potential difference for protons (delta mu approximately H+). The presence of cyanide, the time-independence of the redox potential difference and the irrelevance of the initial redox state of the NAD+/NADH couple ensured that the experimental situation corresponded to a 'static-head condition' with delta mu approximately H+ as the input force and the redox potential difference as the output force, the flow of electrons having reached dynamic equilibrium. Consequently, the observed value of 1.6 for the ratio delta Ge/delta mu approximately H+ is interpreted as indicating that the leads to H+/e- stoicheiometry at 'site 1' is 1.5 and that therefore the mechanism of the proton pump at 'site 1' is not of the group-translocation type (no direct leads to e - leads to H+ coupling). PMID- 6288023 TI - Spin-trapping studies on the free-radical products formed by metabolic activation of carbon tetrachloride in rat liver microsomal fractions isolated hepatocytes and in vivo in the rat. AB - 1. The metabolic activation of carbon tetrachloride to free-radical intermediates is an important step in the sequence of disturbances leading to the acute liver injury produced by this toxic agent. Electron-spin-resonance (e.s.r.) spin trapping techniques were used to characterize the free-radical species involved. 2. Spin trapping was applied to the activation of carbon tetrachloride by liver microsomal fractions in the presence of NADPH, and by isolated intact rat hepatocytes. The results obtained with the spin trap N-benzylidene-2 methylpropylamine N-oxide ('phenyl t-butyl nitrone') (PBN) and [13C]carbon tetrachloride provide unequivocal evidence for the formation and trapping of the trichloromethyl free radical in these systems. 3. With the spin trap 2-methyl-2 nitrosopropane, however, the major free-radical species trapped are unsaturated lipid radicals produced by the initiating reaction of lipid peroxidation. 4. Although pulse radiolysis and other evidence support the very rapid formation of the trichloromethyl peroxy radical from the trichloromethyl radical and oxygen, no clear evidence for the trapping of the peroxy radical was obtainable. 5. The effects of a number of free-radical scavengers and metabolic inhibitors on the formation of the PBN-trichloromethyl radical adduct were studied, as were the influences of changing the concentration of PBN and incubation time. 6. High concentrations of the spin traps used were found to have significant effects on cytochrome P-450-mediated reactions; this requires caution in interpreting results of experiments done in the presence of PBN at concentrations greater than 50 mM. PMID- 6288024 TI - A rapid fluorometric assay for erythrocyte pyridoxal kinase. PMID- 6288026 TI - Binding properties of 23S,25-dihydroxyvitamin D3: an in vivo metabolite of vitamin D3. PMID- 6288025 TI - Tissue distribution of pyrimidine-5'-nucleotidase. PMID- 6288028 TI - Pancreatic phospholipase A2 is not regulated by calmodulin. PMID- 6288029 TI - Insulin enhances protein phosphorylation in isolated hepatocytes by inhibiting an amiloride sensitive phosphatase. PMID- 6288027 TI - 1,25-Dihydroxyvitamin D3 receptor in bovine thymus gland. PMID- 6288030 TI - AGEPC (platelet activating factor) induced stimulation of rabbit platelets: effects on phosphatidylinositol, di- and triphosphoinositides and phosphatidic acid metabolism. PMID- 6288031 TI - GM3 ganglioside induces hamster fibroblast growth inhibition in chemically defined medium: ganglioside may regulate growth factor receptor function. PMID- 6288032 TI - Inhibition of leukotriene B4-induced neutrophil degranulation by leukotriene B4 dimethylamide. PMID- 6288033 TI - Cytochrome C553 from Desulfovibrio vulgaris: potentiometric characterization by optical and EPR studies. PMID- 6288034 TI - Cyclic AMP-dependent phosphorylation of rat liver 6-phosphofructo 2-kinase, fructose 2,6-bisphosphatase. PMID- 6288035 TI - Stimulation of bull seminal Ca2+, Mg2+-dependent endonuclease by various DNA binding proteins. PMID- 6288036 TI - Thyrotropin releasing hormone receptors in regulation of prolactin gene expression in GH cells. PMID- 6288037 TI - Localization of 3,5,3'-L-triiodothyronine receptor in rat kidney mitochondrial membranes. PMID- 6288038 TI - Leukotriene B4 biosynthesis by alveolar macrophages. PMID- 6288039 TI - Electron spin resonance--spin stabilization in enzymatic systems: detection of semiquinones produced during peroxidatic oxidation of catechols and catecholamines. PMID- 6288040 TI - Soluble relaxation factor from vascular smooth muscle: a myosin light chain phosphatase? PMID- 6288042 TI - An active cytochrome c oxidase depleted of subunit III prepared by covalent chromatography on yeast cytochrome c. PMID- 6288041 TI - The influence of temperature and gamma-aminobutyric acid on benzodiazepine receptor subtypes in the hippocampus of the rat. PMID- 6288044 TI - Magnetic field effects on the fluorescence of two reaction centerless mutants of Rhodopseudomonas capsulata. PMID- 6288043 TI - Cannabinoid induced degranulation of rabbit neutrophils. PMID- 6288045 TI - Corticotropin releasing factor stimulates adrenocorticotropin and beta-endorphin release from AtT-20 mouse pituitary tumor cells. PMID- 6288046 TI - Calcineurin is a calmodulin-dependent protein phosphatase. PMID- 6288047 TI - Interaction of the substrate analogue of cytochrome P-450 and mixed function oxidases. AB - The interaction of a spin labeled compound carrying an alkylating group 4-(3-iodo 2-oxopropylidene)-2,2,3,5,5-pentamethylimidozolydene-1-oxyl (RJ) and capable of binding covalently to mixed function oxidases (MFO) was studied. Measurements of the difference spectrum of cytochrome P-450 demonstrated that RJ induces spectral changes characteristic of type I substrates (lambda max = 403 nm; lambda min = 418 nm). The spectral binding constant (Ks) was 66 microM as determined from the difference spectrum. RJ inhibited the microsomal oxidation of substrates of cytochrome P-450 (aniline, aminopyrine and benzo [a]pyrene). This inhibition was shown not to be associated with the conversion of cytochrome P-450 to cytochrome P-420, or with the suppression of the activities of NADPh-cytochrome c reductase and NADPH-cytochrome P-450 reductase. Thus, evidence was obtained for the possible interaction of RJ with cytochrome P-450. RJ injected to rats (5 mg/100 g body wt, i.p.), inhibited the hydroxylation of benzo[a]pyrene, a type I substrate, (21%) and aniline, a type II substrate, (40%) in the microsomes from their livers. The presence of a paramagnetic center in RJ made it possible to study its interaction with microsomes. The electron paramagnetic resonance (EPR) spectrum of RJ was recorded in the rat liver microsomal fraction after in vivo administration of RJ. In rats treated with RJ (5 mg/100 g), hexobarbital sleeping time was prolonged 1.5-fold. Alkylating analogs of substrates of cytochrome P-450 are suggested as agents for structural studies of the active center of cytochrome P-450 and the development of efficient inhibitors of reactions catalyzed by this enzyme. PMID- 6288048 TI - A correlation between hydroxyl radical generation and ethanol oxidation by liver, lung and kidney microsomes. PMID- 6288049 TI - Possible role of microtubules and associated proteases in organophosphorus ester induced delayed neurotoxicity. AB - Organophosphorus delayed neurotoxicants (phenyl saligenin cyclic phosphate and diisopropyl phosphorofluoridate) altered cyclic AMP (cAMP)-dependent phosphorylation and several other processes in brain homogenates and cytoplasmic microtubules. Phenyl saligenin cyclic phosphate slightly stimulated in vitro cAMP dependent phosphorylation in brain homogenates of three species (rat, mouse and rabbit) that have been reported to be insensitive to delayed neurotoxicity, whereas it slightly decreased this phosphorylation in brain homogenates of three sensitive species (chicken, cow and pig) and in brain microtubules of chicken and pig. The microtubule-associated processes that were moderately inhibited by phenyl saligenin cyclic phosphate in sensitive species were: in vitro [3H]cAMP binding to protein kinase, in vitro assembly when tubulin rings were absent, and cAMP-dependent phosphorylation of microtubule-associated proteins (MAPs) both in vitro and on intracerebral administration of 32Pi. The endogenous proteases that degrade the high molecular weight MAPs were strongly inhibited in vitro by phenyl saligenin cyclic phosphate and diisopropyl phosphorofluoridate. In contrast, treatment of chickens with diisopropyl phosphorofluoridate remarkably decreased the in vitro stability of their brain cytoplasmic high molecular weight MAPs, perhaps by enhancing the MAPs-degrading protease activity. These findings indicate that the MAPs-protease system is a possible target for organophosphorus delayed neurotoxicants. PMID- 6288050 TI - Potentiating effects of clofibrate on prostaglandin-dependent and -independent pathways of human platelet activation: evidence for involvement of cyclic AMP. AB - Although clofibrate has been shown to inhibit platelet aggregation that is caused by thrombin, ADP and epinephrine, by blocking the release of arachidonic acid from platelet phospholipids [8], here we have demonstrated that clofibrate enhanced platelet aggregation by arachidonic acid and PLC and reversed the effects of PGE1 on platelet cAMP concentration and on PLC-induced secretion of [14C]-5HT in similar, concentration-dependent manners. Taken together, these findings strongly suggest that the proaggregatory effect of clofibrate is mediated by a lowering of cAMP in platelets. PMID- 6288051 TI - Inhibition of leukotriene biosynthesis in mastocytoma cells by diethylcarbamazine. PMID- 6288052 TI - Brain (Na+,K+)-ATPase: biphasic interaction with erythrosin B. AB - Brain (Na+,K+)-adenosine triphosphatase (EC 3.6.1.3) has both high and low affinity ouabain binding sites. It has been proposed that the high affinity ouabain binding sites characterize a nerve-specific form of the enzyme. Erythrosin B has been reported to inhibit high affinity ouabain binding selectively. The experiments in this paper were carried out in order to characterize the interactions of erythrosin B with (Na+,K+)-ATPase and to examine the specificity of erythrosin B for enzyme with high affinity for ouabain. Inhibition by erythrosin B was biphasic, with a rapid and a slow phase. The rapid phase appeared to be relatively specific for enzyme with high affinity for ouabain, while the slow phase was not. Inhibition by erythrosin B was accelerated by Mg2+ and was retarded by ATP, K+, or Na+ and ATP. Erythrosin B increased apparent affinity of the enzyme for K+ and decreased apparent affinity for Na+ and for ATP. These results indicate that erythrosin B interacts with an ATP site and has effects on cation affinities opposite to those of ATP. Erythrosin B inhibition is proportional to high affinity ouabain binding if brief incubation times and moderate concentrations are used. PMID- 6288053 TI - Inhibition of lymphocyte cyclic AMP phosphodiesterase and lymphocyte function by 5'-methylthioadenosine. PMID- 6288054 TI - Ultrastructure of arthritis induced by a caprine retrovirus. AB - The ultrastructure of early retrovirus-induced arthritis was studied sequentially in 20 goat kids inoculated with caprine arthritis-encephalitis virus. Synovial lesions began as intercellular edema and collagen fragmentation and continued as progressive mononuclear cell infiltration and lining cell hyperplasia, hypertrophy, and necrosis. At 18 through 45 days after the inoculation, lining cells contained small accumulations of virus-like particles similar to virus seen in infected tissue culture cells. No virus was seen budding from infected lining cell membranes. PMID- 6288055 TI - Frequencies of Epstein-Barr virus-inducible IgM anti-IgG B lymphocytes in normal children and children with juvenile rheumatoid arthritis. AB - The relative frequencies of IgM antiIgG autoantibody (rheumatoid factor) producing cells induced by the polyclonal B cell activator Epstein-Barr virus were measured in peripheral blood lymphocyte cultures of normal children and patients with juvenile rheumatoid arthritis. The frequencies of rheumatoid factor precursor B cells in normal children were lower than adults, but higher than neonates. The frequency increased with the age of the donor. In seronegative children with the systemic-onset or pauciarticular-onset types of juvenile rheumatoid arthritis, the number of IgM antiIgG inducible B cells was not significantly different (P greater than 0.05) from age-matched controls. Patients with seropositive juvenile rheumatoid arthritis or seropositive adult rheumatoid arthritis had significantly higher IgM antiIgG precursor cell frequencies than age-matched normal subjects (P less than 0.01 and P less than 0.02, respectively). In contrast, the patients with seronegative polyarticular-onset juvenile rheumatoid arthritis had an average precursor frequency significantly lower than normal age-matched controls (P less than 0.05), analogous to results previously noted in adult seronegative rheumatoid arthritis. Thus, both children and adults with seronegative polyarticular rheumatoid arthritis had a deficiency in B cells that produce IgM antiIgG and that are induced by Epstein-Barr virus. This distinguished them from seropositive juvenile rheumatoid arthritis and rheumatoid arthritis patients, normal subjects, and patients with the pauciarticular-onset and systemic-onset types of seronegative juvenile rheumatoid arthritis. PMID- 6288056 TI - Olecranon bursitis related to calcium pyrophosphate dihydrate crystal deposition disease. AB - A case of olecranon bursitis in an 81-year-old patient is presented. Analysis of the bursal fluid revealed positive birefringent crystals; radiographs showed calcifications in the distal triceps tendon. A bursectomy was performed. X-ray diffraction analyses demonstrated calcium pyrophosphate dihydrate patterns in a subcutaneous "tophus" and in a specimen of the tendon. On histologic examination, there was a bursitis with positive birefringent crystals on the bursa's inner surface; histologic images of "chondrocalcinosis" were observed in and around the tendon. It is concluded that bursitis may be part of the extraarticular manifestations of calcium pyrophosphate dihydrate crystal deposition disease. PMID- 6288057 TI - Studies on CDP-choline:1,2-diacylglycerol cholinephosphotransferase activity in rat arterial wall. AB - The properties of CDP-choline:1,2-diacylglycerol cholinephosphotransferase (CPT) (EC 2.7.8.2.), which catalyzes de novo synthesis of phosphatidylcholine, were studied in rat arterial wall. The optimal pH of CPT of the arterial wall was about 8.5. On subcellular fractionation of the arterial wall, the highest activity was found in the microsome-rich fraction; the cytosolic fraction showed only a trace of activity. The Michaelis constant (KM) for CDP-choline was 0.019 mM. The CPT activity of a homogenate of arterial wall increased linearly with increase in concentration of diolein up to 3.2 mM. 20 mM magnesium and 0.2 mM manganese ions caused marked activation respectively and essential for the activity. Calcium, barium, cobalt, copper, and ferrous ions were inhibitory. 0.5 mM ethylenediaminetetraacetic acid (EDTA) and 0.5 mM glycoletherdiamine-N,N,N'N' tetraacetic acid (GEDTA) increased the activity in the presence of 10 mM magnesium ion. Sonication of the enzyme solution and addition of high concentration of detergent, such as Triton X-100 and Tween 20, markedly decreased the activity. Porcine liver phosphatidylcholine, phosphatidylethanolamine, and especially polyenephosphatidylcholine increased CPT activity of the arterial wall, while lysophosphatidylcholine was strongly inhibitory. The properties of arterial CPT activity under various conditions are discussed. PMID- 6288058 TI - Effects of particle size and perfusate composition on the uptake of colloidal gold by the rabbit thoracic aorta perfused in situ. AB - The influence has been investigated of particle size on the uptake of radioactive gold colloid by the rabbit thoracic aorta perfused in situ. Particles ranging in diameter from 14 nm to 40 nm were suspended in 0.9% NaCl and infused either at a pressure of 15 mm Hg for times of between 2 1/2 and 60 min or at pressure of between 15 and 160 mm Hg for 5 min. Uptake by the whole intima-media increased with perfusion time and hydrostatic pressure but did not depend on particle size. Radioactive assay of serial sections across the aortic wall also showed that particle size did not influence the distribution of tracer. An effect of perfusate composition on uptake was demonstrated in further experiments in which particles either 14 or 40 nm in diameter were suspended in pooled rabbit serum and infused at pressures of between 15 and 140 mm Hg for 5 min. Uptake and transmural distribution were again independent of particle size, but uptake was 4 5-fold less than when the particles were perfused in saline. Under all perfusion conditions radioactivity fell steeply across the intima and then rose gradually across the media and adventitia. Radioactivity in the outer media and adventitia increased with perfusion time but little change could be detected in intimal activity. In transmission electron micrographs, particles in the intima were not seen to penetrate the internal elastic lamella and in the outer media particles remained extracellular and did not enter collagen bundles. Autoradiographs showed that particles in the intima were uniformly distributed around the circumference of the vessel but in the outer media and adventitia particles usually clustered close to the vasa vasorum. PMID- 6288059 TI - [Morphological relations of sarcolemma, cytoplasmic organelles, and nuclei of the junctional and extrajunctional zones]. AB - In intercostal rat muscles an ultrastructural comparison has been made of the relationships existing in the myofibre between sarcolemma, cytoplasmic organules and nuclei in the end-plate sole and extrajunctional areas. Remarkable differences are described. Particularly, in the end-plate sole area relationships between the perinuclear membrane and the floor of the infoldings are very tight. In the extrajunctional areas, close relationships between the portion of perinuclear membrane that is turned towards the interior of the myofibre and the membranous structures of the triad are frequent. PMID- 6288060 TI - [Effect of the oral administration of ranitidine on pituitary secretion]. AB - The acute oral administration of Ranitidine (200 mg.) does not determine significant variations in the plasma levels of PRL, LH, FSH, HGH, ACTH and Cortisol. This fact seems to exclude a vigorous action of the drug, in therapeutic doses, on the hypothalamo-pituitary axis. Furthermore, it cannot be excluded that the chronic administration of Ranitidine cam determine significant variations in the hormones themselves. PMID- 6288061 TI - [Cardiac scintigraphy with technetium 99m pyrophosphate in the diagnosis of a focus of acute myocardial infarction]. PMID- 6288062 TI - Adenylate kinase and malignant hyperpyrexia. PMID- 6288063 TI - Purine pathway enzymes in the circulating malignant cells of patients with cutaneous T-cell lymphoma. AB - The activities of three purine pathway enzymes--adenosine deaminase (ADA), 5' nucleotidase (5'N) and purine nucleoside phosphorylase (PNP)--were examined in the circulating malignant cells (Sezary cells) of eight patients with cutaneous T cell lymphoma (CTCL). Cell lines derived from two other patients with CTCL were also studied. These were compared with enzyme activities in peripheral blood T lymphocytes from 11 normal donors and six samples of human thymocytes. ADA activities were similar in the Sezary cells and peripheral blood T-cells (medians 7 U and 15 U, P = 0.14), and both of these groups demonstrated significantly lower activity than did the thymocytes (median 100 U, P = 0.002). 5'N activity in the Sezary cells was also similar to that of the T-lymphocytes (median 0.022 U and 0.030 U, P greater than 0.05) and both of these groups had significantly greater activity than did the thymocytes (median 0.002 U, P = 0.001). Median PNP activity in the Sezary cell population was also comparable to that measured in normal T-cells. These findings suggest there is a characteristic purine pathway enzyme pattern in Sezary cells that is similar to that seen in normal T lymphocytes. This pattern is clearly distinguishable from that of thymocytes and from that previously described in lymphoblasts from patients with T-cell acute lymphoblastic leukaemia. These results support the concept that Sezary cells are well-differentiated with respect to the T-cell axis. Quantitation of purine pathway enzymes may be useful in defining subsets of T-cell malignancy. PMID- 6288064 TI - Cyclic GMP in urine to monitor the response to ovarian cancer to therapy. AB - Guanosine 3':5'-phosphate (cycle GMP) in urine has been used to monitor the response of patients with ovarian cancer to treatment. Changes in the cyclic GMP level appear to correlate well with clinical status in that the disappearance of clinically detectable tumour is associated with a drop in the level whereas a tumour recurrence is associated with an elevation. Serially measured cyclic GMP is valuable for detecting a recurrence of tumour growth in patients in clinical remission and can predate any clinical signs by as much as 10 months. In patients who show no response to treatment, cyclic GMP levels in urine are elevated in the majority of specimens collected. PMID- 6288065 TI - Isolation of herpes simplex virus from the cornea in chronic stromal keratitis. PMID- 6288066 TI - Secretory IgA specific for herpes simplex virus in lacrimal fluid from patients with herpes keratitis--a possible diagnostic parameter. AB - In the present study a solid-phase radioimmune assay was used for the demonstration of herpes simplex virus-specific IgG and secretory IgA antibodies in the lacrimal fluid from patients with active recurrent herpes keratitis. The method was quantitative and made it possible to test specifically for the production of secretory IgA antibodies produced during an active herpes simplex virus infection. The production of secretory IgA was followed in 2 patients with fresh recurrent lesions. The HSV-specific secretory IgA could be demonstrated during the first 10 days of infection, where the maximal concentration was reached 3-5 days after the first symptoms occurred. The secretory antibodies were locally produced, and it is shown for the first time that herpes virus-specific secretory antibodies were of diagnostic value. PMID- 6288067 TI - The effect of wheat bran on the absorption of minerals in the small intestine. AB - 1. Studies on mineral absorption were carried out in ileostomy patients using the metabolic balance technique. The effect of wheat bran on the absorption of phosphorus, calcium, magnesium, zinc and iron was studied. The extent of digestion of bran phytate in the stomach and small intestine was also investigated. 2. Eight patients with well established conventional ileostomies were studied during two periods while on a constant low-fibre diet. In the second period, 16 g wheat bran/d (American Association of Cereal Chemists) was added to the diet. The amount of phytate-P, non-phytate-P, Ca, Mg, Zn and Fe was determined in the ileostomy contents and in duplicate portions of the diet. 3. Of the added bran phytate-P 24-61% was recovered in the ileostomy contents. In the bran period a significantly decreased amount of Zn was absorbed, while the apparent absorption of Fe and phytate-P increased and that of non-phytate-P, Ca and Mg remained constant. Due to the mineral content of bran, the relative absorption differed in some respects from the absolute absorption, being decreased for Zn, Mg and phytate-P but unchanged for Ca, Fe and non-phytate-P. 4. It is concluded that phytate is partly digested in the stomach and small intestine or possibly absorbed. Addition of 16 g bran/d to the diet does not seem to impair the mineral absorption from the small intestine except that of Zn. PMID- 6288068 TI - Trace element intakes of women. AB - 1. Energy, protein, zinc, copper, manganese, selenium and dietary fibre intakes of 100 pre-menopausal women (mean age 30.0 +/- 6.1 years) from a university community, and consuming self-selected diets, were calculated using 3 d dietary records and food composition values. Subjects also collected a 24 h food composite during the 3 d record period for analysis of Zn, Cu and Mn by atomic absorption spectrophotometry. Daily analysed intakes were compared with those calculated from the corresponding record day. 2. Mean daily calculated intakes of energy, protein, Zn, Cu, Mn, Se and dietary fibre were 7.54 +/- 1.61 MJ, 74 +/- 18 g protein, 10.1 +/- 3.3 mg Zn, 1.9 +/- 0.6 mg Cu, 3.1 +/- 1.5 mg Mn, 131 +/- 53 micrograms Se, 19.4 +/- 6.6 g dietary fibre. 3. Major food sources for each of the trace elements were (%): Zn meat + substitutes 43, dairy products 23.7; Cu breads and cereals 22, vegetables 21; Mn breads and cereals 47, fruits 12, Se meat + substitutes 38, breads and cereals 30. 4. Highly significant correlations (P = 0.001) were noted for analysed intakes of Zn, Cu and Mn and those calculated from the corresponding record day. Mean calculated intakes were higher (%): Zn 138, Cu 142, Mn 121, than corresponding mean analysed intakes (P = 0.01). However, the mean nutrient densities (mg/MJ) were comparable: Zn analysed 1.2, calculated 1.4; Cu analysed 0.2, calculated 0.2; Mn analysed 0.4, calculated 0.4. 5. All subjects met the Canadian Dietary Standard (CDS) recommended level for Cu but 48% received less than the CDS for Zn, 6% obtaining less than two-thirds of this recommended level. Daily Mn and Se intakes were similar to recent values for North American diets. PMID- 6288069 TI - The effects of intraruminal infusions of sodium bicarbonate, ammonium chloride and sodium butyrate on urea metabolism in sheep. PMID- 6288070 TI - The effect of varying the amount of linseed oil supplementation on rumen metabolism in sheep. AB - The effects of three levels of linseed oil (LSO) supplementation of a basal diet on rumen digestion and flow of nutrients to the proximal duodenum of three mature sheep provided with permanent rumen and duodenal re-entrant cannulas were studied. 2. A basal diet of 200 g hay and 400 g concentrates daily, providing approximately 7.0 MJ digestible energy and 13 g N/d, was given alone or with supplements of 13, 26, or 40 ml LSO/d in two equal portions of 06.00 and 18.00 hours. The flow of duodenal digesta was measured by spot-sampling using chromic oxide paper as the marker. Bacterial protein synthesis (BPS) was measured by the diaminopimelic acid technique. 3. Addition of LSO reduced the digestion of energy and organic matter, particularly acid-detergent fibre, in the stomach. Digestion in the intestines increased but at the higher levels of supplementation this failed to compensate completely for the reduction in rumen digestion. Total volatile fatty acid concentrations were not affected but molar proportions of acetate and butyrate were decreased by approximately 18 and 61% respectively while the molar proportion of propionate was increased twofold by the highest concentration of oil. The higher concentrations of LSO virtually eliminated protozoa from the rumen. 4. The second increment of LSO (26 ml/d) produced the highest duodenal flow of total N and bacterial N and the highest efficiency of BPS. The highest concentration of oil (40 ml/d) was without effect. Rumen and duodenal ammonia concentrations and plasma urea concentrations tended to be reduced by the higher concentrations of LSO. 5. It is argued that the results support suggestions made elsewhere that free oils reduce the efficiency of BPS but that they also reduce the numbers of protozoa which can cause an increase in efficiency of BPS. The net effect of free oil supplementation on BPS is thus likely to be variable and difficult to predict. PMID- 6288071 TI - Comparison of changes in inositide and noninositide phospholipids during acute and prolonged adrenocorticotropic hormone treatment in vivo. PMID- 6288072 TI - Rapid-quench and isotope-trapping studies on fructose-1,6-bisphosphatase. AB - Rapid-quench kinetic measurements yielded presteady-state rate data for rabbit liver fructose-1,6-bisphosphatase (FBPase) (a tetramer of four identical subunits) that are triphasic: the rapid release of Pi (complete within 5 ms), followed by a second reaction phase liberating additional Pi that completes the initial turnover of two or four subunits of the enzyme (requiring 100-150 ms), and a steady-state rate whose magnitude depends on the [alpha-Fru-1,6 P2]/[FBPase] ratio. With Mg2+ in the presence of excess alpha-fructose 1,6 bisphosphate (alpha-Fru-1,6-P2) all four subunits turn over in the pre steady state; with Mn2+ only two of the four are active. Thus the expression of half site reactivity is a consequence of the nature of the metal ion and not a subunit asymmetry. In the presence of limiting alpha-anomer concentrations only two of the four subunits now remain active with Mg2+ as well as with Mn2+ in the pre steady state. However, so that the amount of Pi released can be accounted for, a beta leads to alpha anomerization or direct beta utilization is required at the active site of one subunit. Such behavior is consistent with the two-state conformational hysteresis displayed by the enzyme and altered affinities manifested within these states for alpha and beta substrate analogues. Under these limiting conditions the subsequent steady-state rate is limited by the beta leads to alpha solution anomerization. These data in combination with pulse- chase experiments permit evaluation of the internal equilibrium, which in the case of Mg2+ is unequivocally higher in favor of product complexes and represents a departure from balanced internal substrate-product complexes. PMID- 6288073 TI - Nuclease S1 sensitive sites in parental deoxyribonucleic acid of cold-and temperature-sensitive mammalian cells. AB - Temperature-sensitive mutants of 3T3 cells (H6-15) express the transformed phenotype at 33 degrees C and the normal phenotype at 39 degrees C. Cold sensitive mutants of Chinese hamster ovary cells (cs4-D3) express the transformed phenotype at 39 degrees C and the normal phenotype, along with a G1 block, at 33 degrees C. When either cell type is under conditions such that it is normal and in a G0 state, the number of S1-sensitive sites in purified DNA, labeled in parental chains only, is zero. When the normal cells are stimulated by 10% serum, the number of S1 sites per 10(5) base pairs increases slightly, to 0.7 in cs4-D3 and 1.1 in H6-15. Under conditions permitting the expression of the transformed phenotype, but not proliferation, the maximum number of S1 sites per 10(5) base pairs is 5 in cs4-D3 and 44 in H6-15. When the stationary transformed cells are stimulated by 10% serum, the number of S1 sites per 10(5) base pairs increases to 6 in cs4-D3 and 43 in H6-15. Furthermore, the DNA from the stimulated transformed H6-15 cells contains at least twice as many S1 sites as the total number of breaks (nicks plus gaps), raising the possibility of the acquisition of stable looped or cruciform structures as the cells are stimulated. PMID- 6288074 TI - Ultraviolet light induced preferential cross-linking of histone H3 to deoxyribonucleic acid in chromatin and nuclei of chicken erythrocytes. AB - Histones have been cross-linked to DNA in chicken erythrocyte nuclei and chromatin by using ultraviolet light irradiation at 254 nm. Following irradiation, cross-linked histone-DNA adducts were isolated and purified by hydroxylapatite chromatography, and the DNA component was subjected to acid hydrolysis. Of several hydrolysis techniques investigated, trichloroacetic hydrolysis of the DNA component of the adducts was found to be most effective. Histones isolated from hydrolyzed histone-DNA adducts were characterized by gel electrophoresis and fingerprint analysis. No histone-histone protein adducts were observed. All histone fractions have been shown to cross-link DNA in nuclei or chromatin by utilizing the technique employed, but with different propensities. The order of observed cross-linking, deduced from kinetic experiments, is H1 + H5, H3 greater than H4 greater than H2A much greater than H2B. The preferential binding of the core histone H3, as compared to the other core histones, is discussed in light of recent data concerning histone-DNA interactions and nucleosome structure. The use of the ultraviolet light technique as a conformational probe to study chromatin is also discussed. PMID- 6288075 TI - Quaternary-transformation-induced changes at the heme in deoxyhemoglobins. AB - Quaternary-structure-induced differences in both the high- and low-frequency regions of the resonance Raman spectrum of the heme have been detected in a variety of hemoglobins. These differences may be the result of (1) changes in the amino acid sequence, induced by genetic and chemical modifications, and (2) alterations in the quaternary structure. For samples in solution in low ionic strength buffers, differences in the 1357-cm-1 line (an electron-density sensitive vibrational mode) correlate with differences in the 216-cm-1 line (the iron-histidine stretching mode). Thus, changes in the iron-histidine bond and changes in the pi-electron density of the porphyrin depend upon a common heme globin interaction. The quaternary-structure-induced changes in the vibrational modes associated with the heme demonstrate that there is extensive communication between the heme and the globin and impact on models for the energetics of cooperativity. The local interactions of the iron-histidine mode are energetically small and destabilize the deoxy heme in the T structure with respect to the R structure. Therefore, these interactions must be larger in the ligated protein than in the deoxy protein to obtain a negative free energy of cooperativity. Additionally, our data imply that the deprotonation of the proximal histidine does not play a major role in the energetics of cooperativity. On the other hand, models for cooperativity that require conformational changes in the iron-histidine bond or direct interaction between the porphyrin and the protein are qualitatively consistent with the observed variation of heme electronic structure in concert with protein quaternary structure. PMID- 6288077 TI - Physical properties of the glycoprotein of vesicular stomatitis virus measured by intrinsic fluorescence and aggregation. AB - We have studied the denaturation and renaturation of the purified glycoprotein (G) of vesicular stomatitis virus by using intrinsic fluorescence spectroscopy and an aggregation assay. Our studies were carried out with G containing two complex oligosaccharide chains, with the asialo form of the protein, and for some experiments with G containing altered oligosaccharide structures. Fluorescence quenching using acrylamide showed no differences between the native and denatured states of G due to sialic acid content. Denaturation by guanidinium chloride (GdmCl) at 25 degrees C was reversible for the major transition region. The data analyzed by a two-state denaturation model gave a free energy of unfolding in the absence of denaturant of approximately 1.4 kcal/mol. For renaturation, two types of dialysis protocols were employed. The first (direct dialysis) involved dialysis against standard buffer [140 mM NaCl, 10 mM sodium phosphate, 1 mM disodium ethylenediaminetetraacetate, and 0.2% (w/v) poly(oxyethylene) 10 tridecyl ether, pH 7.4]. Recovery of the native emission maximum did not occur for any of the G proteins by using this procedure. The second (annealing dialysis) involved slow removal of GdmCl against decreasing concentrations of GdmCl in standard buffer over a period of 2-3 days. Only in this case was recovery of the native emission maximum and fluorescence intensity obtained. For those G proteins in which the oligosaccharide chains were decreased in size, this protocol led to extensive aggregation. PMID- 6288076 TI - Purification of a skeletal growth factor from human bone. AB - A skeletal growth factor was isolated and purified from demineralized human bone matrix. A dose of 6 micrograms/mL of the purified factor significantly increased the proliferation rate of embryonic chick bone cells in serum-free culture (292% of controls, p less than 0.0001) but had no effect on embryonic chick skin cells plated at the same initial density. The factor is sensitive to inactivation by trypsin and urea, but not by collagenase, 20% butanol, or 1% mercaptoethanol. It is also resistant to inactivation by heat (stable for 15 min at 75 degrees C) and extremes of pH (stable for 30 min at 4 degrees C from pH 2.5 to 10.0). Purification of the active factor by selective heat and acid precipitations, molecular sieve column chromatography, and preparative polyacrylamide gel electrophoresis provided a material that was homogeneous by the criteria of high pressure liquid chromatography, polyacrylamide gel electrophoresis, and isoelectric focusing. The apparent molecular weight is 83 000. The purified factor increases bone cell proliferation at doses comparable to other mitogens: 0.3 microgram/mL (3.6 nM) significantly increases DNA synthesis to 231% of controls (p less than 0.001). The purified factor was also active on cultured embryonic chick bones, enhancing the growth rate of tibiae and femurs, as measured by increased dry weight (185% of controls, p less than 0.025) and [3H]proline incorporation (164% of control, p less than 0.001), respectively. PMID- 6288078 TI - Electron paramagnetic resonance and optical spectroscopic evidence for interaction between siroheme and Fe4S4 prosthetic groups in Escherichia coli sulfite reductase hemoprotein subunit. AB - The hemoprotein subunit (SiR-HP) of Escherichia coli NADPH-sulfite reductase contains one siroheme (high-spin Fe3+, D = 8 cm-1) and one oxidized Fe4S4 center per polypeptide. Christner et al. [Christner, J.A., Munck, E., Janick, P.A., & Siegel, L.M. (1981) J. Biol. Chem. 256, 2098-2101] have shown by Mossbauer spectroscopy that the two prosthetic groups of SiR-HP are magnetically exchange coupled in the oxidized enzyme, a result which indicates the presence of a chemical bridge between them. Photoreduction of SiR-HP in the presence of 5' deazaflavin and ethylenediaminetetraacetic acid causes the enzyme to accept up to 2.0 electrons. The two reducible centers in SiR-HP are reduced independently with a midpoint potential difference of 65 mV, the siroheme being more positive. The first electron added to SiR-HP results in loss of the g = 6.63, 5.24, and 1.98 set of EPR signals due to the ferriheme and production of an EPR-silent state. The second added electron results in the parallel appearance of three distinct types of EPR signal: a novel species with g = 2.53, 2.29, and 2.07 (0.63 spin per heme); two "S = 3/2 type" species with g = 5.23, 2.80, and ca. 2.0 and g = 4.82, 3.39, and ca. 2.0 (together account for 0.16 spin per heme); and a very small amount of a "classical" reduced Fe4S4 center signal with g = 2.04, 1.93, and 1.91 (0.03 spin per heme). The temperature dependences of the "g = 2.29" and "g = 1.93" signals are similar to each other and are like those seen with other Fe4S4 center proteins. Addition of small amounts of guanidinium sulfate (0.1 M) to SiR HP causes the spectrum of fully reduced enzyme to show primarily the S = 3/2 type species (g = 4.88, 3.31, and 2.08; 0.84 spin per heme), although the enzyme remains fully active. Optical spectral changes followed as a function of enzyme reduction show that marked changes occur in the Fe2+ siroheme optical spectrum when the Fe4S4 center becomes reduced or oxidized. These results indicate that the prosthetic groups of SiR-HP remain coupled when the enzyme is reduced. It is suggested that the novel EPR signals result from exchange interaction between S = 1 or 2 ferroheme and S = 1/2 reduced Fe4S4. PMID- 6288079 TI - Properties of the prosthetic groups of rabbit liver aldehyde oxidase: a comparison of molybdenum hydroxylase enzymes. AB - Rabbit liver aldehyde oxidase (AO), like milk xanthine oxidase (XO) and chicken liver xanthine dehydrogenase (XDH), possesses the following prosthetic groups: FAD, a functional Mo center, and two spectroscopically distinct iron-sulfur centers, one with gav less than 2.0 (termed Fe/S I) and the other with gav greater than 2.0 (termed Fe/S II) in the reduced enzyme. EPR spectra for the Mov species were found to be nearly identical in AO and XO for a number of enzyme complexes, and the midpoint reduction potentials for functional MoVI/MoV (-359 mV) and MoV/MoVI (-351 mV) were nearly the same in all three enzymes (50 mM phosphate, pH 7.8). A strong magnetic interaction between MoV and reduced Fe/S I, previously detected in XO and XDH, was also found in AO. No MoV-Fe/S II interaction could be detected in AO (nor in XO). In contrast, the order of reduction of Fe/S I and Fe/S II, as measured from their midpoint potentials, is reversed in AO (Em = -207 and -310 mV, respectively) as compared to XO (Em = -280 and -245 mV, respectively) in phosphate buffer at pH 7.8. The oxidized-reduced extinction coefficients at 450 and 550 nm for the two centers are also apparently reversed in AO and XO. Although magnetic interaction between FAD and one or both reduced Fe/S centers has been detected in both AO and XO, no magnetic interaction between the two reduced Fe/S centers themselves was found in AO (although such interaction has been seen in XO). The average FAD reduction potential is substantially more positive in AO (Em for FAD/FADH., -258 mV; FADH./FADH2, -212 mV at pH 7.8) than in XO or XDH. It can be concluded that although the properties and immediate environment of the functional Mo center are conserved in the three Mo hydroxylase enzymes, and all three enzymes possess the same set of prosthetic groups, the properties of the groups which transfer electrons from the Mo to the ultimate electron acceptor can vary substantially in AO, XO, and XDH. PMID- 6288080 TI - Soluble 5'-nucleotidase: purification and reversible binding to photoreceptor membranes. PMID- 6288081 TI - The reactivity of thiol groups in bovine heart cytochrome c oxidase towards 5,5' dithiobis(2-nitrobenzoic acid). AB - Bovine heart cytochrome c oxidase consists of 12 stoicheiometric polypeptide chains of at least 11 different types. The enzyme contains 14--16 cysteine residues; the distribution of nearly all cysteine residues over the subunits has been established. In native cytochrome c oxidase two thiol groups reacted rapidly and stoicheiometrically with 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB). These thiol groups are located in subunits I and III, respectively. This implies that subunit I is not fully buried in the hydrophobic core of the enzyme. After dissociation of the enzyme by sodium dodecyl sulphate more thiol groups became available to DTNB, in addition to those in subunits I and III, at least one in subunit II, two in fraction V/VI and one to two in the smallest subunit fraction. It is shown that separation of the subunits of cytochrome c oxidase by gel permeation chromatography in the presence of sodium dodecyl sulphate depends on the pH of the elution medium. The elution volume of subunits I, III and VII is dependent on pH, that of the others independent. PMID- 6288082 TI - The pathway of electrons through OH2:cytochrome c oxidoreductase studied by pre steady -state kinetics. AB - The kinetic behaviour of the prosthetic groups and the semiquinones in in QH2:cytochrome c oxidoreductase has been studied using a combination of the freeze-quench technique, low-temperature diffuse-reflectance spectroscopy, EPR and stopped flow. (2) In the absence of antimycin, cytochrome b-562 is reduced in two phases separated by a lag time. The initial very rapid reduction phase, that coincides with the formation of the antimycin-sensitive Qin, is ascribed to high potential cytochrome b-562 and the slow phase to low-potential cytochrome b-562. the two cytochromes are present in a 1:1 molar ratio. The lag time between the two reduction phases decreases with increasing pH. Both the [2 Fe-2S] clusters and cytochrome c1 are reduced monophasically under these conditions, but at a rate lower than that of the initial rapid reduction of cytochrome b-562. (3) In the presence of antimycin and absence of oxidant, cytochrome b-562 is still reduced biphasically, but there is no lag between the two phases. No Qin is formed and both the Fe-S clusters and cytochrome c1 are reduced biphasically, one half being reduced at the same rate as in the absence of antimycin and the other half 10-times slower. (4) In the presence of antimycin and oxidant, the recently described antimycin-insensitive species of semiquinone anion, Qout (De Vries, S., Albracht, S.P.J., Berden, J.A. and Slater, E.C. (1982) J. Biol. Chem. 256, 11996 11998) is formed at the same rate as that of the reduction of all species of cytochrome b. In this case cytochrome b is reduced in a single phase. (5) The reversible change of the line shape of the EPR spectrum of the [2Fe-2S] cluster 1 is caused by ubiquinone bound in the vicinity of this cluster. (6) The experimental results are consistent with the basic principles of the Q cycle. Because of the multiplicity, stoicheiometry and heterogeneous kinetics of the prosthetic groups, a Q cycle model describing the pathway of electrons through a dimeric QH2:cytochrome c oxidoreductase is proposed. PMID- 6288083 TI - The oxidation-reduction kinetics of the reaction of cytochrome c1 with non physiological redox agents. AB - The kinetics of the oxidation-reduction reactions of cytochrome c1 with ascorbate, ferricyanide, triphenanthrolinecobalt(III) and N,N,N',N'-tetramethyl-p phenylenediamine (TMPD) have been examined using the stopped-flow technique. The reduction of ferricytochrome c1 by ascorbic acid is investigated as a function of pH. It is shown that at neutral and alkaline pH the reduction of the protein is mainly performed by the doubly deprotonated form of ascorbate. From the ionic strength-dependence studies of the reactions of cytochrome c1 with ascorbate, ferricyanide and triphenanthrolinecobalt(III), it is demonstrated that the reactions rate is governed by electrostatic interactions. The second-order rate constants for the reaction of cytochrome c1 with ascorbate, ferricyanide, TMPD and triphenanthrolinecobalt(III) are 1.4 . 10(4), 3.2 . 10(3), 3.8 . 10(4) and 1.3 . 10(8) M-1 . s-1 (pH 7.9, I = 0, 10 degrees C), respectively. Application of the Debye-Huckel theory to the data of the ionic-strength-dependence studies of these redox reactions of cytochrome c1 yielded for ferrocytochrome c1 and ferricytochrome c1 a net charge of --5 and --4, respectively. The latter value is close to that of --3 for the oxidized enzyme, calculated from the amino acid sequence of the protein. This implies that not a local charge on the surface of the protein, but the overall net charge of cytochrome c1 governs the reaction rate with small redox molecules. PMID- 6288084 TI - Differential effects of 2,4-dinitrophenol and valinomycin (+ K+) on uncoupler stimulated ATPase of human tumor mitochondria. AB - The uncoupler-stimulated mitochondrial ATPase of four human tumors, mouse kidney, brain and fetal liver exhibited a characteristic behavior when preincubated with the H+-conducting uncouplers, dinitrophenol, CCCP, S-13 and gramicidin. The ATPase activity was considerably lower with preincubation than without. Preincubation with valinomycin (+ K+), on the other hand, did not result in a significant decrease of the ATPase activity. These results may be contrasted with those obtained with liver or heart mitochondria, the ATPase activity of which did not suffer any loss when preincubated with dinitrophenol. The effect of preincubation with dinitrophenol on the tumor mitochondria could not be accounted for by dinitrophenol-induced Mg2+ efflux, since the differential effects of dinitrophenol and valinomycin (+ K+) remained even when ATPase activity was determined in presence of Mg2+. Small amounts of ATP and ADP in the preincubation mixture containing dinitrophenol protected against the decay of the ATPase activity, implicating the exchangeable adenine nucleotides in the tumor mitochondria. In a model system where liver mitochondria were depleted of their adenine nucleotides, a lower ATPase activity was indeed obtained. However, direct determination of the concentrations of adenine nucleotides in dinitrophenol- and valinomycin-treated tumor mitochondria revealed only slight differences. PMID- 6288085 TI - A method for in situ characterization of b- and c-type cytochromes in Escherichia coli and in complex III from beef heart mitochondria by combined spectrum deconvolution and potentiometric analysis. AB - An analytical technique for the in situ characterization of b- and c-type cytochromes has been developed. From evaluation of the results of potentiometric measurements and spectrum deconvolutions, it was concluded that an integrated best-fit analysis of potentiometric and spectral data gave the most reliable results. In the total cytochrome b content of cytoplasmic membranes from aerobically grown Escherichia coli, four major components are distinguished with alpha-band maxima at 77 K of 555.7, 556.7, 558.6 and 563.5 nm, and midpoint potentials at pH 7.0 of 46, 174, -75 and 187 mV, respectively. In addition, two very small contributions to the alpha-band spectrum at 547.0 and 560.2 nm, with midpoint potentials of 71 and 169 mV, respectively, have been distinguished. On the basis of their spectral properties they should be designated as a cytochrome c and a cytochrome b, respectively. In Complex III, isolated from beef heart mitochondria, five cytochromes are distinguished: cytochrome c1 (lambda m (25 degrees C) = 553.5 nm; E'0 = 238 mV) and four cytochromes b (lambda m (25 degrees C) = 558.6, 561.2, 562.1, 566.1 nm and E'0 = -83, 26, 85, -60 mV). PMID- 6288086 TI - Adenine nucleotide pool size, adenine nucleotide translocase activity, and respiratory activity in newborn rabbit liver mitochondria. AB - The adenine nucleotide content (ATP+ADP+AMP) of newborn rabbit liver mitochondria was 6.0 +/- 0.5 nmol/mg mitochondrial protein at birth, increased rapidly to 14.5 +/- 1.7 nmol/mg protein by 2 h postnatal, peaked at 6 h, then decreased gradually to 7.8 +/- 0.6 nmol/mg protein by 4 days postnatal. There was a strong positive correlation (r = 0.82) between the total adenine nucleotide pool size and adenine nucleotide translocase activity in these mitochondria. In contrast, glutamate + malate-supported State 3 respiratory rates remained constant from birth through the first week of life. State 4 rates also remained constant, as did the respiratory control index and uncoupled respiratory rates. The following conclusions are suggested: (1) The maximum rate of translocase activity is limited by the intramitochondrial adenine nucleotide pool size. (2) In newborn rabbit liver mitochondria, the State 3 respiratory rate is not limited by either the adenine pool size or the maximum capacity for translocase-mediated adenine exchange. (3) In contrast to rat, rabbit liver mitochondria are fully functional at birth with regard to respiratory rates and oxidative phosphorylation. (4) The rapid postnatal accumulation fo adenine nucleotides by liver mitochondria, now documented in two species, may be a general characteristic of normal metabolic adjustment in neonatal mammals. PMID- 6288088 TI - The dependence of the conductance of the hemocyanin channel on applied potential and ionic concentration with mono- and divalent cations. AB - Incorporation of Megatura crenulata hemocyanin into phosphatidylcholine black lipid membranes results in the formation of ion channels. Channel properties depend on many factors, three of which are examined in this work: type and concentration of electrolyte and applied voltage. Eight cations at different concentrations have been used. Instantaneous conductance of the channel is a saturating function of both applied voltage and ionic strength of the bathing solution with monovalent cations, but only of ionic strength with divalent cations. Steady-state voltage-conductance relations are nonlinear for both signs but show slight saturation with ionic strength. Relaxation towards the steady state can be fitted by two exponentials with different time constants. All experimental data are fitted postulating the existence of a mechanism of voltage gating of the channel, and of discrete negative charge near its mouth. Specific and nonspecific binding of cations is required. PMID- 6288087 TI - The site of inhibition by 5,5'-dithiobis(2-nitrobenzoate) in ubiquinol: cytochrome c oxidoreductase. AB - In 5,5'-dithiobis(2-nitrobenzoate) (DTNB)-treated succinate: cytochrome c reductase, the electron transfer from duroquinol to cytochrome c is inhibited due to the fact that the Rieske Fe-S cluster and, consequently, cytochrome, c, are no longer reducible by substrate. The finding that, after this treatment, cytochrome b is still reducible by substrate in the absence of antimycin, but not in its presence, is consistent with a Q-cycle mechanism for the electron transfer through QH2:cytochrome c oxidoreductase. The inhibitory effect of DTNB and its effect on the EPR spectrum of the [2Fe-2S] cluster suggest that it prevents either the binding of ubiquinone in the vicinity of this cluster or the interaction between the Fe-S protein and a ubiquinone-binding protein. PMID- 6288089 TI - Comparison of red cell and kidney (Na+ +K+)-ATPase at 0 degrees C. AB - Human red cell and guinea pig kidney (Na+ +K+)-ATPase were phosphorylated at 0 degrees C. Using concentrations of ATP ranging from 10(-6) to 10(-8) M, ATP dependent regulation of reactivity is observed with red cell but not kidney (Na+ +K+)-ATPase at 0 degrees C. In particular, with the red cell enzyme only, the following are observed: (i) the ratio of enzyme-bound ATP (E.ATP, measured by the pulse-chase method of Post, R.L., Kume, S., Tobin, T., Orcutt, B. and Sen, A.K. (1969) J. Gen. Physiol. 54, 306s-326s) to steady-state level of total phosphoenzyme (EP) decreases with decrease in ATP concentration and (ii) the apparent turnover of phosphoenzyme (ratio of Na+-stimulated ATP hydrolysis to level of total EP at steady state) also varies as a function of ATP concentration. In addition, when EP is formed at very low ATP (0.02 microM), and then EDTA is added, rapid disappearance of a fraction of EP occurs, presumably due to ATP resynthesis, only with the red cell enzyme. These differences in behaviour of the red cell and kidney enzymes are explained on the basis of the observed predominance of K+-insensitive EP in red cell, but K+-sensitive EP in kidney (Na+ +K+)-ATPase at 0 degrees C. PMID- 6288090 TI - Localization of (Ca2+ + Mg2+)-ATPase, Ca2+ pump and other ATPase activities in cardiac sarcolemma. AB - N-Ethylmaleimide was employed as a surface label for sarcolemmal proteins after demonstrating that it does not penetrate to the intracellular space at concentrations below 1.10(-4) M. The sarcolemmal markers, ouabain-sensitive (Na+ +K+)-ATPase and Na+/Ca2+-exchange activities, were inhibited in N-ethylmaleimide perfused hearts. Intracellular activities such as creatine phosphokinase, glutamate-oxaloacetate transaminase and the internal phosphatase site of the Na+ pump (K+-p-nitrophosphatase) were not affected. Almost 20% of the (Ca2+ +Mg2+) ATPase and Ca2+ pump were inhibited indicating the localization of a portion of this activity in the sarcolemma. Sarcolemma purified by a recent method (Morcos, N.C. and Drummond, G.I. (1980) Biochim. Biophys. Acta 598, 27-39) from N ethylmaleimide-perfused hearts showed loss of approx. 85% of its (Ca2+ +Mg2+ ATPase and Ca2+ pump compared to control hearts. (Ca2+ +Mg2+)-ATPase and Ca2+ pump activities showed two classes of sensitivity to vanadate ion inhibition. The high vanadate affinity class (K1/2 for inhibition approx. 1.5 microM) may be localized in the sarcolemma and represented approx. 20% of the total inhibitable activity in agreement with estimates from N-ethylmaleimide studies. Sucrose density fractionation indicated that only a small portion of Mg2+-ATPase and Ca2+ ATPase may be associated with the sarcolemma. The major portion of these activities seems to be associated with high density particles. PMID- 6288091 TI - Ouabain-insensitive Na+-stimulated ATPase activity of basolateral plasma membranes from guinea-pig kidney cortex cells. II. Effect of Ca2+. AB - The ouabain-insensitive, Mg2+-dependent, Na+-stimulated ATPase activity present in fresh basolateral plasma membranes from guinea-pig kidney cortex cells (prepared at pH 7.2) can be increased by the addition of micromolar concentrations of Ca2+ to the assay medium. The Ca2+ involved in this effect seems to be associated with the membranes in two different ways: as a labile component, which can be quickly and easily 'deactivated' by reducing the free Ca2+ concentration of the assay medium to values lower than 1 microM; and as a stable component, which can be 'deactivated' by preincubating the membranes for periods of 3-4 h with 2 mM EDTA or EGTA. Both components are easily activated by micromolar concentrations of Ca2+. The Ka of the system for Na+ is the same, 8 mM, whether only the stable component or both components, stable and labile, are working. In other words, the activating effect of Ca2+ on the Na+-stimulated ATPase is on the Vmax, and not on the Ka of the system for Na+. The activating effect of Ca2+ may be related to some conformational change produced by the interaction of this ion with the membranes, since it can also be obtained by resuspending the membranes at pH 7.8 or by ageing the preparations. Changes in the Ca2+ concentration may modulate the ouabain-insensitive, Na+-stimulated ATPase activity. This modulation could regulate the magnitude of the extrusion of Na+ accompanied by Cl- and water that these cells show, and to which the Na+ ATPase has been associated as being responsible for the energy supply of this mode of Na+ extrusion. PMID- 6288092 TI - Inhibition of the Na+/K+ cotransport system by cyclic AMP and intracellular Ca2+ in human red cells. AB - Human erythrocytes are able to incorporate cyclic AMP (cAMP) in amounts larger than those required to saturate cAMP-dependent protein kinase. In contrast to previous observations in avian red blood cells in which cAMP stimulates the Na+/K+ cotransport system, we demonstrate that cAMP inhibits this system in human erythrocytes. The cotransport inhibition is enhanced by addition of phosphodiesterase inhibitor 1-methyl-3-isobutylxanthine to the incubation medium. The cAMP concentration giving half-maximal cotransport inhibition showed a wide variation among different individuals (from 0.1 to 5 mM external cAMP concentration). In contrast to cAMP, cyclic GMP showed little effect on the cotransport system. Ca2+ introduced into the cell interior was an inhibitor of the Na+/K+ cotransport system. These results suggest that in human cells in which endogeneous levels of cAMP and Ca2+ are modulated by hormones, the Na+/K+ cotransport system may be under hormonal regulation. PMID- 6288094 TI - The H+/ATP stoichiometry of the (H+ +K+)-ATPase of dog gastric microsomes. AB - Gastric microsomal vesicles isolated from dog fundic mucosa were shown to be relatively ion tight and have a low level of proton permeability. The H+ translocase, basal ATPase and K+-activated ATPase activities of these vesicles were measured and the H+/ATP stoichiometry calculated using either the total K+ ATPase or the K+-stimulatable component (total K+-ATPase--basal ATPase). The former estimations consistently gave stoichiometric of approximately one, whereas the use of only the K+-stimulatable component gave widely differing values. Measurement of the dephosphorylation of the enzyme under basal conditions revealed both a labile and a stable phosphoenzyme component. The rate of decay of the labile component completely accounted for the basal ATPase activity observed. We conclude that the basal ATPase associated with our preparations is a spontaneous dephosphorylation of the phosphoenzyme occurring in the absence of K+ and that the H+/ATP stoichiometry of the gastric ATPase is one. PMID- 6288093 TI - Electrical measurement of electroneutral fluxes of divalent cations through charged planar phospholipid membranes. AB - The voltage-sensitive channel-former monazomycin is used as a conductance probe to monitor changes in the trans electrostatic surface potentials of negatively charged planar phospholipid bilayers. Cis-to-trans electroneutral fluxes of divalent cations mediated by ionophores A23187 and X537A are sensed via the effect of transported divalent cations on the trans surface potentials. Quantitative determinations of neutral Ca2+ and Mg2+ fluxes are made and related to ionophore function. PMID- 6288095 TI - Interaction of hemoglobin with the red blood cell membrane. A saturation transfer electron paramagnetic resonance study. AB - Human hemoglobin has been labeled on cysteine 93(beta) with the maleimide spin label, 3-maleimido-2,2,5,5-tetramethyl-1-pyrrolidinyloxyl and reassociated with erythrocyte membrane previously stripped of hemoglobin and glyceraldehyde-3 phosphate dehydrogenase. The affinity of hemoglobin for the membrane is not affected by the presence of the label. Saturation transfer electron paramagnetic resonance measurements show that the diffusion rotational movements of hemoglobin are considerably slowed down when it is bound to the erythrocyte membrane. The correlation time of rotation, tau c, is found to be 8 . 10(-6) s as compared with 2 . 10(-8) s when the hemoglobin molecule is in solution. The same values are obtained whether the protein is associated with its high- or low-affinity binding sites. They depend on the viscosity of the solution. The high-affinity sites are presumably located on the segment of the band 3 protein which extends into the cytoplasm and which links through ankyrin, the spectrin-actin cytoskeleton to the membrane. When band 3 is cross-linked into a dimer after reaction with the copper ortho-phenanthroline chelate, the correlation time of rotation of spin-labelled hemoglobin is unchanged. It is also independent of the presence of the spectrin actin network and ankyrin. These results show tha the movements of hemoglobin bound by ionic linkage to different part (protein or phospholipid) of the cytoplasmic surface of the membrane are similarly highly restricted by some potential or energetic barrier. They give also evidence for independent movements and flexibility in the assembly of the macromolecules which link the spectrin actin cytoskeleton to the erythrocyte membrane. PMID- 6288096 TI - Multiple effects of sulphydryl reagents on sugar transport by rat soleus muscle. AB - Iodoacetate, over the range 0.2-2 mM, stimulated the uptake of D-xylose by rat soleus muscle and inhibited anaerobic lactate production by soleus muscle. Stimulation of sugar transport is considered to be due to the resultant fall in ATP. p-Chloromercuribenzene sulphonate (0.5-2 mM) stimulated xylose uptake to a lesser extent than iodoacetate and induced a proportionately smaller fall in ATP, consistent with the inhibitory effect of p-chloromercuribenzene sulphonate on lactate production. Under certain conditions, p-chloromercuribenzene sulphonate stimulated sugar transport without affecting the ATP level. This suggests that whereas p-chloromercuribenzene sulphonate can be expected to stimulate sugar transport through the lowering of muscle ATP, it may also act through some other mechanism. No stimulatory effect on xylose uptake was observed when muscles were exposed to N-ethylmaleimide (0.02-2 mM) either for brief (1 min) or more prolonged (30 min) periods. Because N-ethylmaleimide induced a marked fall in muscle ATP, it is surprising that N-ethylmaleimide did not stimulate sugar transport; in most experiments this inhibitor actually inhibited sugar transport. N-Ethylmaleimide inhibited the stimulation of sugar transport by 2,4 dinitrophenol and anoxia; this inhibitory effect appears to explain why N ethylmaleimide itself did not stimulate sugar transport. p-Chloromercuribenzene sulphonate also inhibited 2,4-dinitrophenol-stimulated xylose uptake by a mechanism which seems similar to that of N-ethylmaleimide; this could explain in part the modest stimulatory effect of this inhibitor on muscle sugar transport. PMID- 6288097 TI - The relationship between plasma membrane lipid composition and physical-chemical properties. III. Detailed physical and biochemical analysis of fatty acid substituted EL4 plasma membranes. AB - Murine leukemia EL4 cells were modified by supplementation of culture media with fatty acids for 24 h. A plasma membrane-enriched fraction was prepared from substituted and normal cells. Analyses were performed to determine fatty acyl composition, phospholipid headgroup composition and cholesterol content. The two major membrane phospholipids, phosphatidylethanolamine (PE) and phosphatidylcholine (PC) were isolated by thin-layer chromatography and ESR measurements were done on liposomes prepared from these lipids as well as on the intact plasma membrane preparations. Slight perturbations in overall plasma membrane lipid composition were observed when EL4 cells were supplemented with a single exogenous fatty acid. This may be consistent with the idea that the incorporation of exogenous fatty acid induces compensatory changes in membrane lipid composition. On the other hand, we observed no significant difference in two ESR motional parameters between the unsubstituted control and various fatty acid-substituted plasma membranes. ESR measurements carried out on PE and PC liposomes derived from 17:0- and 18:2c-substituted membranes also failed to detect major differences between these liposomes and those made from normal EL4 phospholipids. In the case of liposomes prepared from 18:2t,-substituted membranes, the order parameter was significantly changed from the normal. However, the change was in opposite directions in PE and PC, perhaps accounting for the fact that no change parameter is seen in intact 18:2t-substituted plasma membrane. Measurements of order parameter (S) in mixed lipid vesicles showed that at up to 50 mol% mixture of a synthetic PC with plasma membrane PC, the value of S was only marginally different from that of the plasma membrane PC vesicles. We interpret these data as an indication that the two ESR parameters used are not sufficiently sensitive to detect changes due to modifications of the acyl chain composition of a complex biological membrane. PMID- 6288098 TI - Stimulation of transepithelial sodium and chloride transport by ascorbic acid. Induction of Na+ channels is inhibited by amiloride. PMID- 6288099 TI - A comparison of the mobilities and thermal transitions of retrovirus lipid envelopes and host cell plasma membranes by electron spin resonance spectroscopy. AB - The lipid bilayers of several type-C retroviruses and selected host cells were spin labeled with 5-doxyl stearic acid, and intact viruses and cells were subjected to electron spin resonance spectroscopy in order to measure lipid mobility. Thermal transition profiles generated for four different retroviruses were dissimilar; differences in the values of the hyperfine splitting constant 2T parallel and in the positions of thermal break points reflect variations in mobility which can be correlated with the phospholipid/cholesterol molar ratios of the viral envelopes. Moreover, removal of virion surface projections by protease digestion altered the mobility of the envelope and in the positions of thermal break points, but the effect observed depended upon the particular retrovirus examined. Studies on retrovirus-infected and uninfected host cells have revealed that persistent virus infection can elicit changes in host plasma membrane mobility and in the positions of thermal break points, the direction and magnitude of which are highly dependent upon the particular retrovirus-host cell system under consideration. PMID- 6288100 TI - Protein synthesis in human lymphoblastoid cells (Namalwa) after treatment with butyrate and 5'-bromodeoxyuridine. AB - The pattern of protein synthesis has been compared in Namalwa cells following treatment with butyrate and 5'-bromodeoxyuridine (BrdUrd). Although these treatments cause a substantial increase in Sendai virus-induced interferon synthesis (up to 300-fold) we observed no comparable effect on the synthesis of other cellular proteins. Using two-dimensional gel electrophoresis we have investigated the proteins synthesised before and 8 h after Sendai virus infection of treated cells. Only 2 of the 300 most abundant cellular proteins were reproducibly affected, these always showed increased rates of synthesis in butyrate-treated cells. The most significant was a 3-4-fold enhancement in synthesis of a 35,000 molecular weight protein which we have called BEP35. On individual occasions treatment caused changes in the rates of synthesis of other proteins, these were not reproducible and involved less than 4% of the proteins investigated. None of the Sendai virus structural proteins or virus-induced cellular proteins were affected by the treatment. We conclude that butyrate and BrdUrd treatments have a relatively specific effect on the synthesis of interferon in Namalwa cells, as the majority of protein synthesis remains unaffected. PMID- 6288101 TI - Chromatin structure interferes with excision of abnormal bases from DNA. AB - Cell-free extracts of human lymphoblastoid cells NL3 excised almost all uracil residues from free DNA with misincorporated dUMP, but only about half the uracil residues from nuclei, chromatin and reconstituted chromatin with dUMP misincorporated DNA. This difference in susceptibility to uracil-DNA glycosylase of free and complexed DNAs was similar to the difference in susceptibility of free and complexed methylated DNAs to 3-methyladenine-DNA glycosylase. Methylated poly(dA-dT) was also protected by formation of complexes with calf thymus chromosomal proteins. It seems that the nucleosome structure prevents the action of DNA glycosylases. The very high sensitivity of PBS1 phage DNA, which contains uracil as a natural component, in complexes with calf thymus chromosomal proteins as well as in the free form [1] was confirmed. This high sensitivity seems ascribable to the high uracil content of PBS1 DNA. Methylated nucleosome monomers and dimers, and reconstituted nucleosome monomers containing methylated DNA of about 150 bp length, were considerably more resistant to 3-methyladenine-DNA glycosylase than chromatin reconstituted from methylated DNA of longer chain length. This may be due to the lower proportion of linker regions of free form stretches of the DNA chain in nucleosome oligomers. PMID- 6288102 TI - The purification of human DNA fragments containing benzpyrene adducts. AB - DNA was isolated from human diploid lung epithelial cells treated in culture with [3H]benzpyrene. The DNA contained one covalently bound benzpyrene group per 38 kb and it was digested with a series of restriction endonucleases giving decreasing fragment sizes, and also with DNAase I to 96% acid solubility. The digests were applied to octyl-Sepharose columns under conditions which promote hydrophobic interaction of the benzpyrene groups on the DNA with the octyl groups in the column matrix. Separation of fragments without and with benzpyrene groups was achieved with successive high salt and ethanediol washes. As DNA fragment size is decreased, more DNA-associated benzpyrene is eluted with ethanediol. Under these conditions, DNA from untreated cells is totally removed in the high salt wash and free benzpyrene metabolites are retained on the column. The separation of DNA fragments with covalently-bound hydrophobic benzpyrene groups, from less modified or unmodified DNA will facilitate examination of the distribution of benzpyrene adducts in defined regions of the human genome. PMID- 6288103 TI - Altered expression of ribokinase activity in Novikoff hepatoma variants. AB - Ribokinase, the first enzyme in ribose catabolism, is altered in its expression in ribose-utilizing Novikoff hepatoma variants. 90% of the variants selected for their ability to use D-ribose as a sole carbon source show a change in ribokinase activity. After non-selective growth, phenotypically unstable variants lose their altered expression and regain a parental form of expression of this enzyme. In the variants, ribokinase expression in non-inducible by the carbon source and is unaffected by the growth phase of the cells. However, ribokinase expression in both parental cells and variants is cell cycle-dependent. Parental Novikoff hepatoma cells have three peaks of ribokinase activity during the S, G2 and M phases. Variants are described which have high basal levels of ribokinase and only a single peak of enzymatic activity during the S phase. In addition to changes in the level of ribokinase, changes in the subcellular localization of the enzyme have been found in some variants. While the change in the level of ribokinase seems to be a property of the variant isolated, the change in subcellular location of ribokinase can be readily achieved by culture conditions. PMID- 6288104 TI - Dephosphorylation of glycogen synthase from human polymorphonuclear leukocytes. AB - Leukocyte glycogen synthase (UDPglucose:glycogen 4-alpha-d-glucosyltransferase, EC 2.4.1.11) was phosphorylated to about one P1/synthase subunit by either the cAMP-dependent protein kinase or the cAMP-dependent synthase kinase. The relationship between dephosphorylation and the increase in the ratio of independence was investigated by analysis of the release of 32P-labelled phosphopeptides from the trypsin-sensitive and the trypsin-insensitive regions. The trypsin-insensitive region was predominantly dephosphorylated and a close correlation between dephosphorylation of a phosphopeptide in the trypsin insensitive region and activation of glycogen synthase is reported for the enzyme phosphorylated in both ways. PMID- 6288105 TI - Selective alteration of Ca2+-dependent and Ca2+-independent cyclic nucleotide phosphodiesterase activity in rat cerebral cortex by cyclic nucleotides and their analogs. AB - The effects of various cyclic nucleotides and cyclic nucleotide analogs on the activity of Ca2+-independent and Ca2+-dependent phosphodiesterases purified from rat cerebral cortex were examined. The order of potency for inhibition of the Ca2+-dependent enzyme by the various agents was the same using either cAMP or cGMP as substrate with 2'-O-monobutyryl cGMP, cIMP and 2-deoxy cGMP being the most potent. The inhibition of deoxy cGMP using cAMP or cGMP as substrate was competitive, with Ki values of 11 and 13 microM, respectively. In marked contrast, hydrolysis of cAMP or cGMp by the Ca2+-independent enzyme was stimulated 50-75% by cIMP, deoxy cGMP and N2-monobutyryl cGMP with EC50 values of 7, 20 and 30 microM, respectively. cGMP (EC50, 1.5 microM) produced quantitatively the same degree of stimulation of cAMP hydrolysis by the Ca2+ independent phosphodiesterase and the activation was not additive with that of deoxy cGMP. Of the other derivatives examined, 2'-O-monobutyryl cAMP and 2'-deoxy cAMP were the most potent inhibitors of cAMP hydrolysis by the Ca2+-independent enzyme and were 30-60 times more effective in inhibiting cAMP hydrolysis as compared to cGMP hydrolysis. The specificity of K+ in inhibiting the activity of the Ca2+-dependent phosphodiesterase and deoxy cAMP in inhibiting the Ca2+ independent enzyme may provide convenient means to examine specifically these activities in crude extracts from rat cerebral cortex. PMID- 6288106 TI - High-affinity 86Rb-binding and structural changes in the alpha-subunit of Na+,K+ ATPase as detected by tryptic digestion and fluorescence analysis. AB - High-affinity 86Rb-binding has been related to tryptic cleavage and fluorescence from intrinsic and extrinsic probes in order to examine the relationship of cation binding to structural transitions in the alpha-subunit of pure membrane bound Na+,K+-ATPase from the outer renal medulla. Native Na+,K+-Atpase binds two Rb+ ions per alpha-subunit (12.3 nmol/mg protein) with high affinity (Kd = 7.5 microM) in 25 mM Tris-HCl, pH 7.5. Enzyme with one molecule of covalently attached fluorescein per alpha-subunit has the same capacity (12.8 nmol/mg protein) but a much lower affinity for Rb+ (Kd = 29.2 microM). The changes in conformational state of the protein are correlated with occupancy of the high affinity sites for Rb+, also at concentrations of Rb+ below the Kd. Titration at varying ionic strength suggests that the E2-form is the relaxed or native conformation of the alpha-subunit. Changes in tryptic digestion pattern and in fluorescence are parallel events both in the conditions of the binding assay and at physiological ionic strength. Reversible blocking of sulfhydryl groups with Thimerosal (ethylmercurythiosalicylate) abolishes the fluorescence responses to K+ or Rb+ without affecting the capacity or the affinity for binding of 86Rb. The demonstration of high-affinity binding of Rb+ without coupling to a conformational change suggests that the E1-form of the protein exposes sites for tight binding of K+ or Rb+ at the cytoplasmic membrane surface. PMID- 6288107 TI - The interaction of Mn2+ with turkey erythrocyte adenylate cyclase. AB - Mn2+ at concentrations below 0.1 mM supports the activation of turkey erythrocyte adenylate cyclase (ATP pyrophosphate-lyase (cyclizing), EC 4.6.1.1) by beta agonists or NaF, similarly to Mg2+ at 5.0 mM. At higher concentrations, Mn2+ is strongly inhibitory, as is Mg2+ above 6 mM. Also, Mn2+ with GTP, but in the absence of beta-agonist, is very potent in reversing the Gpp(NH)p permanently active state to the basal state. beta-Receptor (R) to guanyl nucleotide regulatory protein (N) coupling still occurs at inhibitory Mn2+ concentrations, since the intrinsic kinetic parameters which characterize the R to N coupling interrelationship are unaffected by Mn2+ at a wide concentration range. It is suggested that the inhibitory effect of Mn2+ is due to the impairment of the guanyl nucleotide regulatory protein (N) to the catalytic subunit (C) interaction. PMID- 6288108 TI - Effect of vanadate on the formation and stability of the phosphoenzyme forms of 2,3-bisphosphoglycerate-dependent phosphoglycerate mutase and of phosphoglucomutase. AB - 2,3-Bisphosphoglycerate-dependent phosphoglycerate mutase (2,3-bisphospho-D glycerate:2-phospho-D-glycerate phosphotransferase, EC 2.7.5.3) and phosphoglucomutase (alpha-D-glucose-1,6-bisphosphate:alpha -D-glucose-1-phosphate phosphotransferase, EC 2.7.5.1), which are markedly inhibited by vanadate, possess a ping-pong mechanism involving an intermediate phosphoenzyme. The formation and the stability of these phosphoenzymes have been examined spectrophotometrically in the absence of vanadate. Vanadate does not inhibit the phosphorylation of either mutase by its cofactor. The instability of the phosphoenzyme form of phosphoglycerate mutase increases in the presence of vanadate, but the stability of the phosphorylated phosphoglucomutase is not affected. PMID- 6288109 TI - Chain elongation and desaturation of eicosapentaenoate to docosahexaenoate and phospholipid labeling in the rat retina in vivo. AB - The metabolism of [1-14C]eicosapentaenoic acid in the retina after intravitreal injections in the adult rat eye was studied. The acylation of eicosapentaenoic acid and the appearance of labeled docosapentaenoate and docosahexaenoate in individual phospholipids was observed at 3, 5 and 30 min after injection. The elongation and desaturation products represented about 8 and 4%, respectively, of the total radioactivity of phospholipids 3 min after injection. The highest labeling was found in phosphatidylcholine, phosphatidylinositol and phosphatidic acid. The uneven labeling profiles and the specific activities in individual phospholipids suggested that, in addition to the deacylation-acylation route for the introduction of polyenoic acyl groups into phospholipids, acylation may also take place during the synthesis of phosphatidic acid, followed by channeling to phospholipids. PMID- 6288110 TI - Extraction and detergent/lipid activation of dolichol kinase. AB - The CTP-dependent dolichol kinase from bovine liver microsomes was optimally extracted using either 0.5% sodium deoxycholate or 0.5% Triton X-100 containing 0.5 M NH4Cl. All activity was found in the supernatant fraction following high speed centrifugation. This fraction was depleted of phospholipid (phospholipid remaining, less than 5% of total) by gel chromatography of the 0.5% deoxycholate extract. This partially purified enzyme was maximally activated 9- or 53-fold over controls in the presence of 0.1% deoxycholate or 0.1% Triton X-100, respectively. Stimulation of the kinase was also observed with mixtures of dimyristoylphosphatidylcholine and deoxycholate. The level of stimulation by these mixtures was up to 20-fold higher than that observed in controls having deoxycholate alone. Dimyristoylphosphatidylcholine alone was not stimulatory. A 1:1 molar ratio of Triton X-100 or deoxycholate to dimyristoylphosphatidylcholine was optimal for enzyme activation. The half-maximum velocity of the dephospholipidated enzyme at 1:1 molar ratio of detergent to dimyristoylphosphatidylcholine was obtained at 150 or 550 microM CTP in the presence of deoxycholate or Triton X-100, respectively. It has been observed, therefore, that dolichol kinase may be extracted from liver microsomes, depleted of endogenous phospholipids and activated by specific molar ratios of detergent to phospholipid. PMID- 6288111 TI - Characterization of the interaction between gastrin and its receptor in rat oxyntic gland mucosa. AB - A gastrin receptor, identified in crude membrane preparations of rat oxyntic gland mucosa, has an equilibrium dissociation constant (Kd) of approx. 4 . 10( 10)M and a binding capacity of 4 fmol/mg protein. The binding capacity was significantly lower after 2 days of fasting, parallel with a significant drop in serum gastrin levels; there was no change in Kd. In order to verify Scatchard analysis and to determine if there was a coincident alteration in the association (k+1) and dissociation (k-1) rates in the fasted rat, a kinetics study was performed. Under our conditions, there appeared to be a single set of binding sites and the binding reaction obeyed first-order dissociation, and second-order association rate kinetics. Second-order association rate kinetics were validated by demonstrating the independence of the rate constants when there were alterations in the concentrations of reactants. The average k+1 was determined to be 2 . 10(6) M-1 . s-1. The average k-1 was determined to be 1 . 10(-3) s-1. There was no significant change in the k+1 and k-1 in fed and fasted rats. Fasting decreased the number of gastrin receptors without altering the affinity of the receptor for the hormone. PMID- 6288112 TI - Demonstration of distinct forms of testicular adenylate cyclases associated with germinal and Leydig cell fractions. AB - Decapsulated testes from adult rats were digested with collagenase, and the fraction enriched in germinal and Leydig cells was applied to a 0-4% continuous metrizamide gradient and centrifuged. This leads to separation of a germinal cell fraction and two putative Leydig cell populations that bind human choriogonadotropin, but only one of which responds to the gonadotropin with marked increase in testosterone production. Adenylate cyclase activity was present in these three fractions, and Mn2+ was more effective than Mg2+ as a divalent cation. The adenylate cyclase activity associated with the germinal cell fraction was just marginally stimulated by fluoride and by the non-hydrolyzable GTP analog 5'-guanylimidodiphosphate, while that associated with the Leydig cell populations was stimulated to a greater degree depending upon the type of divalent cation. Only the Leydig cell populations exhibited marked human choriogonadotropin-sensitive stimulation of adenylate cyclase activity in the presence of 5'-guanylimidodiphosphate above that observed with the GTP analog alone. These results suggest the presence of distinct adenylate cyclases in adult rat testis and indicate that both populations of Leydig cells are capable of producing cyclic AMP in response to gonadotropins such as human choriogonadotropin. PMID- 6288113 TI - Formation of sorbitol 6-phosphate by bovine and human lens aldose reductase, sorbitol dehydrogenase and sorbitol kinase. AB - Formation of sorbitol 6-phosphate by bovine and human lens aldose reductase and sorbitol dehydrogenase by the reduction of glucose 6-phosphate and fructose 6 phosphate, respectively, has been demonstrated. The reaction product has been identified by Dowex-formate column chromatography, gas chromatography and mass spectrometry. Sorbitol 6-phosphate can also be formed by the phosphorylation of sorbitol by lens sorbitol kinase in the presence of ATP. PMID- 6288114 TI - Effect of retinoic acid and 12-O-tetradecanoyl phorbol-13-acetate on the binding of epidermal growth factor to its cellular receptors. AB - Binding of 125I-labelled epidermal growth factor (EGF) to C3H/2K cells and the effect of a tumor promotor, 12-O-tetradecanoyl phorbol-13-acetate (TPA) and of a tumor promotor antagonist, retinoic acid, on the binding was studied. Scatchard plot analysis of the binding showed the presence of two types of binding sites with different affinity to EGF. Treatment of the cells with retinoic acid for 1 h resulted in elevation of the affinity of both sites without changing their number per cell. Prolonged exposure to retinoic acid abrogated this elevation of the affinity and caused cycloheximide-sensitive increase of the number of the binding sites of both types. TPA inhibited binding of EGF to the cells by abolishing the binding to the high affinity sites, whereas retinoic acid, in the presence of TPA, enhanced it by increasing the number of the low affinity sites. PMID- 6288115 TI - Biochemical characterization of the OXI mutants of the yeast Saccharomyces cerevisiae. AB - OXI mutants in Saccharomyces cerevisiae lack a functional cytochrome c oxidase. Wild type and OXI mutants were grown in the presence of radioactive delta amino[14C]levulinic acid, a precursor of porphyrin and heme, and [3H]mevalonic acid, a precursor of the alkyl side-chain of heme a. SDS polyacrylamide gel electrophoresis of the delipidated mitochondria showed that delta amino[14C]levulinic acid was distributed into three bands migrating in the regions of Mr 28 000, 13 500, and 10 000, while [3H]mevalonic acid was found in a single band with apparent Mr of 10 000. The immunoprecipitates obtained by incubating the solubilized mitochondria of any OXI mutant with antibodies against cytochrome c oxidase, showed, after delipidation, a high specific radioactivity due to delta-amino[14C]levulinic acid and [3H]mevalonic acid. This suggested that a prophyrin a was present in all these OXI mutants. HCl fractionation confirmed the presence of porphyrin a in the apooxidase of these mutants. Atomic absorption spectra of the immunoprecipitate of cytochrome c oxidase showed that copper was not detectable in the mutant OXI IIIa which lacked subunit 1, but was present in the mutant OXI IIIb, which exhibited a minor alteration in the electrophoretic mobility of subunit 1. In OXI I and II mutants there was a 50% reduction in the amount of copper in the immunoprecipitated cytochrome c oxidase. These observations may be interpretable as follows: (1) alterations in polypeptide biosynthesis due to the OXI mutations lead to an improper configuration of cytochrome c oxidase, so that ferrochelatase cannot transfer iron into porphyrin a; (2) subunit I is the binding site for copper, but the mutations in subunits II and III alter the binding site of one of the two copper atoms in subunit I. PMID- 6288116 TI - Hyperoxic-induced alterations in lung cell structure and function. Effects on cellular cyclic AMP content and comparison to alterations produced by exogenous cyclic AMP. AB - Hyperoxic exposure in vitro of two lung-derived cell types (the epithelial derived L2 cells and WI-38 fibroblasts) inhibits cellular replication, produces striking morphologic changes and may result in cell death; these effects have been observed consistently in other cell types. Hyperoxic exposure of L2 cells is associated with an increase in cellular cyclic AMP content (cellular cyclic AMP content 454 +/- 115 fmol/micrograms DNA in cells exposed to pO2 677 Torr for 96 h compared to 136 +/- 17 fmol/microgram DNA in air-grown cells). Hyperoxic exposure of WI-38 fibroblasts is not associated with increased cyclic AMP content. Although cultivation of L2 cells in the presence of exogenous dibutyryl cyclic AMP does inhibit replication and produce morphologic alterations, similar effects are produced by sodium butyrate alone. Hyperoxic exposure alters cyclic AMP metabolism in some cell types, but the structural and functional alterations observed in L2 cells and WI-38 fibroblasts following hyperoxic exposure are not produced by changes in cellular cyclic AMP content. PMID- 6288117 TI - Effects of products from macrophages, blood mononuclear cells and or retinol on collagenase secretion and collagen synthesis in chondrocyte culture. AB - Collagenase secretion was studied on cultures of rabbit articular chondrocytes. Differentiation of the cells was assessed by characterizing the type of 3H labelled collagen produced during treatment with (1) conditioned media from rabbit peritoneal macrophages and human blood mononuclear cells, and (2) with retinol, a potent cartilage resorbing agent in tissue culture. Conditioned media stimulated collagenase secretion. Total collagen synthesis was reduced due to a decrease of synthesis of alpha 1 chains; the amount of alpha 2 chains synthesized was unchanged. This is thought to be due to a reduction in type II synthesis. Retinol did not stimulate collagenase secretion. Total collagen synthesis was reduced by retinol. alpha 2 chain synthesis, however, was significantly increased, suggesting a switch of collagen synthesis in favor of type I collagen, and therefore, dedifferentiation. These results demonstrate that dedifferentiation of chondrocytes with respect to collagen synthesis is not necessarily associated with a stimulation of collagenase secretion. PMID- 6288118 TI - Cytogenetical and biochemical characterization of a dG + dC-rich satellite DNA in the primate Cebus capucinus. AB - A very abundant and dG + dC rich DNA satellite from primate Cebus capucinus has been characterized in its cytogenetic and biochemical properties with the purpose of studying the correlation between the staining properties of heterochromatin and the base composition of the corresponding very repetitive DNA. The staining techniques, as well as incorporation of base analogues, show that the heterochromatin segments of C. capucinus chromosomes correspond to a dG + dC-rich satellite. This satellite was detected and isolated by centrifugation in density gradient, radioactively labelled and localized by in situ hybridization on heterochromatin segments. PMID- 6288119 TI - Fixation of aromatic hydrocarbons to proteins and DNA mediated by superoxide radical and other activated oxygen species. PMID- 6288120 TI - BVP models: an adjustment to express a mechanism of inactivation. AB - Fitzhugh's BVP model has been used by many people. Fitzhugh has pointed out that as the stimulus is increased the model has "inverted" behaviour. It is here shown that this is due to a lack of a mechanism of inactivation, and the model is adjusted by supplying such a mechanism, to give a new model, called BPH, which, like Fitzhugh's, is of second order, with one equation of first degree and the other of third degree in the dynamical variables. Numerical solutions of the new system are compared with numerical solutions of the Hodgkin-Huxley and of the Fitzhugh equations. PMID- 6288121 TI - [Partial purification and properties of cGMP-dependent protein kinase from prawn tissue]. AB - Using ion-exchange chromatography and gel filtration, cGMP-dependent protein kinase was purified from prawn tissues 220-fold with a yield of activity of 12%. The apparent Ka values for cGMP, cAMP and 8-Br-cGMP are 1 . 10(-7), 5 . 10(-6) and 5 . 10(-8) M, respectively; the apparent Km values for ATP in the presence of cGMP is 9 . 10(-6) M. The cGMP-stimulated protein kinase activity was observed only in the presence of SH-compounds and high Mg2+ concentrations (500-100 mM). The protein kinase demonstrated a broad pH optimum wih a maximum at pH 6.8-7.2. The elution volume of the enzyme during gel filtration corresponded to a globular protein with molecular weight of 140,000. PMID- 6288122 TI - [Effect of transformed steroids on Na,K-dependent ATPase]. AB - The transformed steroids containing delta 5-3 beta-hydroxy- or 3 beta, 5 alpha dihydroxy-6-keto groups in the A/B rings and an additional cycle E (17,20 dihydroxy-delta-lactone, 16,23-pyranone or delta 20(22)-16 alpha, 17 alpha dihydroxy-23-carbethoxy-side chain) (1 . 10(-5) M) inhibit Na+,K+-ATPase from pig kidney medulla or from ox brain. The steroid structure has a noticeable effect on ATPase inhibition varying from 3 to 26%. The data obtained suggest that the uneven distribution of polar substituents in the steroid molecule is essential for ATPase inhibition by steroid aglycons. PMID- 6288123 TI - [Structure-function organization of ACTH: fragment ACTH 11-24--a functionally important site of the hormone molecule]. AB - The steroidogenic and lipolytic activities of ACTH fragments (ACTH11-24--I, ACTH11-19--II, ACTH11-16--III and ACTH 17-24--IV) were studied. Fragments I--IV exert a steroidogenic effect in isolated fasciculata rat adrenal cells at concentrations of 1--500 micrograms/ml. The inner activity (alpha) and concentration at which a half-maximum effect is achieved (EC50) for fragments I and IV are 0.64+/-0.09 and 0.5--2.0 micrograms/ml, for fragment III--0.49+/-0.07 and 0.7 microgram/ml, respectively. Fragments I--IV have no effect on the lipolysis in isolated rat fat cells. The results obtained are indicative of the functional importance of fragment ACTH11-24 in manifestation of steroidogenic action of ACTH and suggest that the second active site of ACTH is enclosed within this amino acid sequence. PMID- 6288125 TI - [Effect of benzimidazole derivatives and levamisole (phenylimidothiazole) on cyclic 3':5'-AMP phosphodiesterase from pig brain]. AB - The effects of phenylimidothiazole and some benzimidazole derivatives on phosphodiesterase of cyclic 3':5'-AMP (cAMP) from pig brain were studied. Depending on their concentrations the compounds tested activated or inhibited the enzyme activity. A correlation between the effects of benzimidazoles and phenylimidothiazole which is known to exert an immunomodulating action was observed. PMID- 6288124 TI - [Role of CO-binding cytochrome c in enzymatic oxidation of methane by the bacterium Methylococcus capsulatus]. AB - The cytochrome c spectrally related to cco cytochromes has been isolated and purified from the methane-oxidizing bacterium Methylococcus capsulatus. The cytochrome binds CO but does not bind other substrates of methane monooxygenase, does not activate the methane monooxygenase reaction and is not a component of methane monooxygenase. In the methanol dehydrogenase enzymatic system cytochrome cco functions as electron acceptor. A possible role of cytochrome cco as electron carrier intermediate in the sequence of the dehydrogenase and oxidase enzymatic systems of M. capsulatus is discussed. PMID- 6288126 TI - Changes in cerebrospinal fluid cyclic nucleotides in alcohol-dependent patients suffering from delirium tremens. AB - In eight patients with a history of alcoholism (on average 9 years), cyclic 3',5' monophosphate (AMP) and cyclic 3',5'-monophosphate (GMP) were determined in CSF in the acute untreated phase of delirium tremens and at 2 weeks later when clinical symptoms had vanished. Before the second lumbar puncture a drug-free period of 1 week had existed. The results were compared with an age- and sex matched neurological control group. CSF cyclic AMP concentrations were markedly reduced by 62% (p less than 0.005) in the acute state of delirium tremens and remained at the same level 2 weeks later; cyclic GMP concentrations were increased by 37% (p = 0.05) and showed a small further increase (p less than 0.05) at the second lumbar puncture. A negative correlation existed between age and cyclic AMP content of CSF (r = -0.756; p less than 0.05) in the patients group. The data indicate that the earlier observed increase in norepinephrine turnover in the acute state of delirium tremens (Athen et al., 1977) seems not to induce an increase of cyclic AMP content in CSF. PMID- 6288128 TI - Modulation of pituitary luteinizing hormone releasing hormone receptors by sex steroids and luteinizing hormone releasing hormone in the rat. PMID- 6288127 TI - Effects of cholinergic and GABAergic drugs on apomorphine-induced gnawing behavior in rats. AB - The effects of cholinergic and GABAergic agonist and antagonist were studied on the apomorphine-induced gnawing behavior in rats. Physostigmine (0.5 mg/kg) decreased the biting scores, whereas atropine (5 mg/kg) had an opposite effect. Picrotoxin potentiated the physostigmine inhibitory action on gnawing responses. This inhibitory gnawing effect was antagonized by prior treatment with atropine. Pretreatment with amino-oxyacetic acid (25 mg/kg) also potentiated the inhibitory effect of physostigmine on gnawing behavior in animals. Such an additive inhibitory response could be antagonized by simultaneous pretreatment with cholinergic and GABAergic blockers such as atropine and picrotoxin, respectively. These findings indicate that the modulatory action of cholinergic neurons on DA system is probably influenced by changes in the GABAergic system. PMID- 6288129 TI - Cyclic adenosine monophosphate-dependent protein kinases of human seminal plasma: origin and characteristics of multiple forms. PMID- 6288130 TI - In vitro effects of cannabinoids on follicular function in the rat. AB - delta 1-Tetrahydrocannabinol (delta 1-TCH), the major psychoactive constituent of marihuana, was found to suppress the preovulatory surge of gonadotropins and thereby to prevent ovulation in rats, rabbits and rhesus monkeys. These studies suggested that the drug acts primarily on the hypothalamus to suppress luteinizing hormone releasing hormone (LHRH) secretion. The aim of the present study was to examine the direct effect of delta 1-THC, the psychoactive constituent of marihuana and cannabidiol (CBD), one of its nonpsychoactive constituents, on preovulatory rat follicles in vitro. Both cannabinoids inhibited follicular steroidogenesis in a dose-dependent manner. Basal accumulation of progesterone (P), testosterone (T) and estradiol-17 beta (E2) was reduced up to 60% by the highest doses examined (100-200 microM). The luteinizing hormone (LH) stimulated increase in P and T was inhibited by 75-88% by the highest doses of both cannabinoids (50-200 microM), while E2, accumulation was inhibited by only 40%. It appears that the inhibitory action of cannabinoids is exerted beyond LH binding and activation of adenylate cyclase and prior to pregnenolone formation in the gonadal steroidogenic pathway. In addition to this anti-steroidogenic effect, both cannabinoids induced resumption of meiosis in follicle-enclosed oocytes cultured in hormone-free medium; 200 microM delta 1-THC resulted in 80% maturation and CBD in 75%. It seems that the action of cannabinoids on rat follicles in vitro is unrelated to their psychotropic activity. PMID- 6288131 TI - Short-term primary culture of mouse interstitial cells: effects of culture conditions of androgen production. AB - This study examined the effects of culture conditions and hormone treatment on androgen production by mouse interstitial cells in short-term primary culture. Testicular interstitial cells (18-25% 3 beta-hydroxysteriod dehydrogenase positive) were maintained in serum-free hormone supplemented medium. Basal (nonstimulated) androgen production was found to be plating-density dependent. Androgen production per cell increased dramatically in a time- and cell concentration-dependent manner. This effect was reproduced in low density cultures by addition of charcoal-stripped conditioned medium from high density cultures. The cell anchorage factors, fibronectin and poly-l-lysine, similarly enhanced basal androgen production but did not augment responsiveness to luteinizing hormone (LH). Coating of the culture surface with serum inhibited androgen production. Cultured cells remained responsive to LH for 4 to 5 days and both insulin (5 micrograms/ml) and epidermal growth factor (EGF) (3 ng/ml) augmented LH-stimulated androgen production. There was a transient increase in LH sensitivity and maximum LH-stimulated androgen production for 5 to 72 h in culture followed by a decline in androgen production to low levels after 4 to 5 days in culture. This loss of activity was partially prevented by addition of antioxidants to the medium or by reduction of the ambient O2 concentration to 1%. PMID- 6288132 TI - Electrostatic modeling of ion pores. Energy barriers and electric field profiles. AB - This paper presents calculations of the image potential for an ion in an aqueous pore through lipid membrane and the electric field produced in such a pore when a transmembrane potential is applied. The method used is one introduced by Levitt (1978, Biophys. J. 22:209), who solved an equivalent problem, in which a surface charge density is placed at the dielectric boundary. It is shown that there are singularities in this surface charge density if the model system has sharp corners. Numerically accurate calculations require exact treatment of these singularities. The major result of this paper is the development of a projection method that explicitly accounts for this behavior. It is shown how this technique can be used to compute, both reliably and efficiently, the electrical potential within a model pore in response to any electrical source. As the length of a channel with fixed radius is increased, the peak in the image potential approaches that of an infinitely long channel more rapidly than previously believed. When a transmembrane potential is applied the electric field within a pore is constant over most of its length. Unless the channel is much longer than its radius, the field extends well into the aqueous domain. For sufficiently dissimilar dielectrics the calculated values for the peak in the image potential and for the field well within the pore can be summarized by simple empirical expressions that are accurate to within 5%. PMID- 6288133 TI - Proton longitudinal relaxation investigation of histidyl residues in human normal adult hemoglobin. AB - The longitudinal relaxation of the C2 protons of surface histidyl residues as well as other aromatic protons of human normal adult deoxyhemoglobin investigated at 360 MHz is discussed in terms of the theory proposed by Kalk and Berendsen for the proton longitudinal relaxation in proteins (Kalk, A., and H.J.C. Berendsen. 1976. J. Magn. Reson. 24:343-366). The role of the four paramagnetic iron atoms of deoxyhemoglobin as fast-relaxing sinks for the overall proton longitudinal relaxation is evaluated according to the model proposed by Bloembergen for the relaxation of nuclei in crystals containing paramagnetic centers (Bloembergen, N. 1949. Physica. 15:386-426). The results suggest that the effectiveness of the paramagnetic iron atoms of deoxyhemoglobin for the overall proton longitudinal relaxation is reduced as a result of slower spin diffusion and wide distribution of methyl groups within the hemoglobin molecule. Thus, deoxyhemoglobin provides a good model for investigating the influence of cross relaxation on proton longitudinal relaxation in proteins at the slow motion limit and in the presence of paramagnetic centers. For the C2 protons of surface histidyl residues, we show that the cross relaxation resulting from the interresidue dipolar interaction makes an important contribution to their longitudinal relaxation. PMID- 6288134 TI - Three-dimensional disorder of dipolar probes in a helical array. Application to muscle cross-bridges. AB - Fluorescence polarization and EPR experiments on azimuthally randomized helices bearing extrinsic (dipolar) probes yield information about the axial orientation and order of the probes. If the orientation of the probe on the structure bearing it is known and disorder is absent, the orientation of the structure may be ascertained. For cases where less probe orientation information is available and/or disorder is present, the available structural information is correspondingly reduced. Here we examine the available data on probes attached to cross-bridges in muscle fibers: four plausible cases of three-dimensional cross bridge disorders are numerically modeled muscle in states of rigor and relaxation. In rigor, where the reported probe disorder is small (Thomas and Cooke, 1980), it was found that the cross-bridge disorder was also small. On the other hand, for the relaxed state where the probes are found to be completely disordered, the cross-bridges may have a considerable amount of order. This possibility is in concert with the results of x-ray diffraction, in which the presence of well-developed myosin-based layer lines indicates considerable order in relaxed muscle. PMID- 6288135 TI - Orientation relaxation of DNA restriction fragments and the internal mobility of the double helix. AB - The orientation relaxation of 15 DNA restriction fragments (43-4361 base-pairs) is characterized by measurements of linear dichroism using high electric field pulses. The off-field relaxation of fragments of 84 base-pairs or less can be described by single exponentials, which are related to the transverse rotational diffusion of the helix. Fragments of 95 base-pairs or greater exhibit an additional fast component with time constants around 100 ns for fragments of approx. 100 base-pairs, increasing with chain length to about 700 ns for a fragment with 258 base-pairs. The amplitude of this process increases from virtually zero at low fields (approximately equal to 10 kV) to a substantial limit contribution at high fields. According to these results, we suggest that electric fields induce stretching of the DNA fragments from a weakly bent to a more straight form and that the fast component reflects the internal mobility of the DNA chain. The slow off-field components of the orientation are discussed in terms of different models. The data up to helix lengths of about 400 base-pairs can be described by the 'weakly bending rod' model from Hearst using 3.4 A rise per base-pair and 13 A axial radius of the helix. Both the weakly bending rod according to Hearst and the 'wormlike chain' according to Hagerman and Zimm provide a persistence length of 500 A. The on-field relaxation is slower than the corresponding off-field process at low field strengths, but the on-field process is accelerated substantially at high electric fields. These observations are compared with model calculations of Schwarz. PMID- 6288136 TI - A small DNA restriction fragment with an apparent permanent dipole moment. AB - The electric birefringence of two DNA restriction fragments, each containing 147 base-pairs, has been investigated. The decay of the birefringence was the same for the two fragments, with a relaxation time corresponding to the reorientation of fully extended, rod-like molecules. However, the birefringence saturation behavior of the two fragments was markedly different: one fragment oriented by the expected induced-dipole mechanism, while orientation of the other fragment followed the theoretical curve for permanent dipole orientation. This difference in behavior must be due to differences in the base-pair sequences of the two fragments. PMID- 6288137 TI - Single channel gating events in tracer flux experiments. II. Flux amplitude analysis. AB - Measurement of tracer ion flux from or into a collection of closed membrane structures (CMS) constitutes a broadly applicable technique for studying ion channel gating by specialized gating molecules in biological membranes. The amplitudes for the flux process reflect the overall change in tracer content due to flux during a period in which channels on at least some of the CMS were open. In practice, the attainment of a time-invariant, finite overall tracer content, indicating a cessation of flux, need not imply that flux has reached completion, i.e., that the CMS internal and external tracer concentrations have fully reached equilibrium. Less than maximum flux amplitudes arise when binding of control ligands leads to an inhibition or inactivation of the channel gating molecules prior to a complete equilibration of tracer. Analysis of the dependence of the flux amplitudes on control ligand concentration permits determination of characteristic parameters of the CMS that may vary with the methods of preparation (e.g., the distributions of CMS size and CMS content of gating units). Knowledge of these parameters in turn permits evaluation of the mean single channel flux amplitude contribution, which is functionally dependent on the rate constant ratio (k'eff/ki), where k'eff and ki are, respectively, the effective rate constants for tracer flux and for gating unit inactivation. PMID- 6288138 TI - Single channel gating events in tracer flux experiments. III. acetylcholine receptor-controlled Li+ efflux from sealed Torpedo marmorata membrane fragments. AB - Filter assay measurements of Li+ efflux from acetylcholine receptor-containing vesicular Torpedo marmorata membrane fragments (microsacs) are presented. Techniques are introduced for: (a) inducing a complete emptying of the Li+ content of all microsacs containing one or more functionally intact receptors, and (b) for determining the distribution of internal volumes of the microsacs using filtration with membrane filters of different pore sizes. The flux amplitudes resulting for acetylcholine receptor-controlled Li+ efflux, when receptors are inhibited by alpha-bungarotoxin or inactivated by a neuroactivator induced desensitization process, were measured. Amplitude analysis was used to determine characteristic parameters of the microsacs that may vary with the technique of preparation (e.g., the distribution in size and receptor content), as well as the mean single channel flux amplitude contribution (e-kt)infinity, which represents the mean reduction of the Li+ content of a microsac due to efflux from a single receptor-controlled channel closing due to inhibition or inactivation of the receptor. The ratio keff/ki was found to lie in the range 0.1 less than keff/ki less than 0.5, where keff and ki are, respectively, the rate constant for Li+-Na+ exchange flux and for the slow inactivation reaction mode of the acetylcholine receptor induced by carbamoylcholine at high concentrations. PMID- 6288139 TI - Columns of water molecules and chloride ions in hydrophobic channels in crystals of a potassium complex. PMID- 6288140 TI - Mammalian cells contain a DNA-recombining activity that can be dissociated from topoisomerase activity. AB - The role of the topoisomerase enzyme in DNA recombination was investigated by extracting chromosomal deoxyribonucleoproteins from a variety of cultured mammalian cells and assaying for the formation of recombinant DNA structures. Although each of the crude deoxyribonucleoprotein preparations contained topoisomerase activity, they did not all contain DNA-recombining activity. A distinct, perhaps novel, enzyme may therefore promote DNA recombination in these cell-free systems. PMID- 6288141 TI - Concepts related to adaptation of neuromuscular function and craniofacial morphology. PMID- 6288142 TI - [Role of cyclic nucleotide phosphodiesterase in the effect of estradiol on uterine tissue]. AB - It is shown that 17 beta-estradiol (10(-7)--10(-5) M) inhibited phosphodiesterase activity of the preparations (supernatant 100000 epsilon, 1 h) obtained from uterine tissue of sexually mature rats and did not affect adenylate cyclase activity of crude membrane fraction of this tissue. The hormone did not change phosphodiesterase activity of the preparations obtained from the brain, heart and outer segments of the retinal rods. Cytosol preparations from uterine tissue were demonstrated to be able to specific hormone binding. The antiestrogen clomifen completely blocked the binding. In the presence of clomifen estradiol had no effect on phosphodiesterase activity. It is suggested that estrogen receptors are necessary for the effect of 17 beta-estradiol on phosphodiesterase to be realized in uterine tissue. PMID- 6288143 TI - [Effect of a varying corticosteroid allowance on the cAMP content of limbic system structures of the rat brain]. AB - A study was made of the effect of adrenalectomy and substitution therapy in the absence of adrenals on the cAMP content in the hypothalamus, hippocamp and striate body of the rat brain. It was shown that in sham adrenalectomized animals, the concentration of cAMP in the brain structures under study did not differ from the characteristics recorded in the control group. Adrenalectomy in rats was followed by a significant reduction of the nucleotide in all the structures, namely in the hypothalamus by 89.5%, and in the hippocamp and striate body by 30.3 and 30.5%, respectively. Application of substitution therapy (5 mg hydrocortisone per 100 mg body mass) prevented the decreased content of cAMP seen in the adrenalectomized rats, with its magnitude being within the limits recorded in the intact animals. Doca was little effective in this aspect. PMID- 6288144 TI - Extrarenal erythropoietin production by macrophages. AB - Murine bone marrow and adherence-separated spleen cells cultured on hydrophobic, gas-permeable Teflon foils (petriperm dishes) can be shown to synthesize and secrete erythropoietin (Epo) and colony-stimulating activity (CSA) simultaneously into the surrounding medium. The Epo activity in the supernatants of primary cultures as measured by the fetal liver erythroid colony-forming technique, from adherent and nonadherent spleen cells, increases over the first 7 days in culture, followed by a plateau until 14 days. Use of the macrophage-specific cytotoxic agent, crystalline silica, as a tool to release residual Epo contained in these cells produces a similar time-Epo activity curve to that found in the primary supernatants. This, together with functional and morphological examination of the cells, indicates that macrophages are responsible for this activity. The total Epo activity released from adherent and nonadherent spleen cells at plateau levels was estimated to be 25 mU/ml culture/day. Weekly subcultivation of bone marrow and adherence-separated spleen cells initiated from primary cultures demonstrated a massive increase in both Epo activity and CSA above that obtained for the primary cultures. Subcultivation could be continued for at least 6 wk. These results, together with the reversible inhibition of Epo and CSA production by cycloheximide, demonstrate that these molecules are synthesized by the macrophage. The evidence supports the hypothesis that the macrophage is involved not only in extrarenal Epo production, but also in the possible short-range regulation of hemopoiesis. PMID- 6288145 TI - Elevated NAD(P) glycohydrolase activity: a possible enzymatic marker for malignancy in Burkitt's lymphoma cells. AB - A comparative analysis of enzymatic activities has been performed on 47 human continuous lymphoid lines: 22 tumors derived from Burkitt's lymphoma lines, 6 other lymphomatous long-term cultures, and 19 nonmalignant ties determined on the cell extracts. 4 showed no significant differences between the various lines. They included adenosine diphosphoribose incorporation, glucose-6-phosphate dehydrogenase, cyclic-AMP phosphodiesterase, and glutathione reductase. However, striking differences of activity were found for the enzyme, NAD(P) glycohydrolase (EC 3.2.2.6). Activity levels were, as a mean, four times higher in Burkitt's lymphoma-derived cell lines than in nonmalignant control lines, and the difference was highly significant (p less than 0.02). All Burkitt cell lines containing translocations of chromosome 8 with either chromosomes 2, 14 or 22 showed an increased activity. The specificity and significance of this possible enzymatic marker of Burkitt's lymphoma cells is discussed. PMID- 6288146 TI - Evidence for interaction between platelet fibrinogen receptors. AB - Previous analysis of fibrinogen binding to human aspirin-treated gel-filtered platelets yielded upwardly concave Scatchard plots. To ascertain whether this was due to the presence of independent heterogeneous receptor populations binding fibrinogen with different affinities, the dissociation of purified 125I fibrinogen from ADP-treated gel-filtered platelets was evaluated as a function of receptor occupancy. Dissociation of bound labeled fibrinogen was measured after 50-fold dilution with buffer containing O, 0.2, 0.8, and 2.0 mg/ml unlabeled fibrinogen. Dissociation of labeled fibrinogen increased with increasing receptor occupancy and was biphasic. With buffer alone, 76.0% +/- 5.8% (SD) of labeled fibrinogen dissociated in 30 min, with an initial rate of 0.392 +/- 0.175 min-1; with 0.2 mg/ml fibrinogen, 83.7% +/- 3.9% dissociated, with an initial rate of 0.589 +/- 0.044 min-1; with 0.8 mg/ml, 91.8% +/- 1.3% of the labeled fibrinogen dissociated, with an initial rate of 0.910 +/- 0.028 min-1; and with 2.0 mg/ml fibrinogen, 97.3% +/- 2.3% of label dissociated, with an initial rate of 1.06 +/- 0.257 min-1 (n=5). The final rates of fibrinogen dissociation were unaffected by unlabeled fibrinogen in the dilution buffer and were not statistically different from the final dissociation rate of 0.015 +/- 0.10 min-1 observed following dilution with buffer alone. These results were neither an artifact of aspirin treatment or gel filtration, as similar observations were made using non-aspirin treated washed platelets, nor were they an artifact of the purified fibrinogen preparations, because binding studies using whole plasma as the major source of fibrinogen also yielded upwardly concave Scatchard plots. Since the data demonstrate that the initial rate and extent of fibrinogen dissociation are dependent on fibrinogen receptor occupancy, they suggest receptor interactions possibly resulting from receptor clustering or crosslinking. Because the dissociation was biphasic, the results also suggest some heterogeneity among platelet-fibrinogen interactions. PMID- 6288148 TI - Functional and non-functional nerve-smooth muscle transmission in the renal arteries of the newborn and adult rabbit and guinea-pig. AB - Isolated renal arteries from newborn and adult guinea-pigs failed to respond to electrical stimulation of the intramural nerves, using parameters identical to those which elicit large neurogenic contractions in renal arteries from newborn and adult rabbits. The threshold stimulation frequency was lower and responses were greater in adult compared with neonatal rabbits. Comparisons of noradrenaline EC50 values showed that guinea-pig renal arteries are significantly less sensitive to noradrenaline that rabbit vessels. Quantitative fluorescence histochemistry of the perivascular adrenergic nerves in renal arteries revealed that the density of innervation gradually declines in the guinea-pig but substantially increases in the rabbit during postnatal development. The lack of neurogenic response of the guinea-pig renal artery, despite the existence of a varicose, adrenergic nerve plexus, is discussed. PMID- 6288147 TI - Responses to adenine nucleotides and related compounds of isolated dog cerebral, coronary and mesenteric arteries. AB - In helically cut strips of dog cerebral, coronary and mesenteric arteries contracted with prostaglandin F2 alpha, adenosine, AMP, ADP, ATP and cyclic AMP produced dose-related, persistent relaxation. On the basis of ED50S, the relaxant effect seen in cerebral arteries was in the order of AMP, ADP, ATP greater than adenosine, cyclic AMP greater than aderine, dibutyryl cyclic AMP greater than inosine. On the basis of maximum relaxations, the effect was in the order of adenine, dibutyryl cyclic AMP greater than or equal to adenosine, AMP, ADP, ATP greater than cyclic AMP, inosine. The relaxant responses to adenosine, AMP, ADP, ATP and cyclic AMP were attenuated by treatment with aminophylline, whereas the relaxations induced by dibutyryl cyclic AMP, adenine and inosine were not altered. It is concluded that adenosine, AMP, ADP and ATP, but not adenine and inosine, appear to share receptors underlying arterial relaxations, and the cyclic AMP-induced relaxation does not derive from increments in cellular cyclic AMP. Treatment with aspirin did not alter the relaxant effect of ADP, adenosine and adenine, suggesting that the release of vasoactive prostaglandins is not involved. PMID- 6288149 TI - Association studies of polybrominated biphenyls in aquatic systems. PMID- 6288150 TI - The treatment of unicameral or solitary bone cysts by the injection of corticosteroids. AB - Methylprednisolone acetate injections into bone cysts are as effective a treatment as curettement and packing. More than one injection may be necessary since recurrences have been seen in half the cases treated with a single injection. Fracture of the cystic part may occur before or after injection if bony filling has not taken place. The fracture may be serious if it involves the neck of the femur. The method is safe and does not result in an unsightly scar. Solitary eosinophilic granulomata of bone also reportedly respond positively to direct injections fo methylprednisolone into the lesion by signs of healing and with relief of pain. PMID- 6288151 TI - Valvular heart disease associated with juvenile onset ankylosing spondylitis: a case report and review of the literature. AB - Aortitis with associated aortic insufficiency is a well-known extra-articular manifestation of ankylosing spondylitis and other HLA-B27 associated spondyloarthropathies in adults. The cardiovascular histopathology is distinct from other forms of inflammatory and degenerative valve disease. Clinical cardiac involvement may be slowly progressive or fulminant and life-threatening. Mitral valve involvement associated with juvenile onset ankylosing spondylitis has rarely been reported. This article discusses a case of juvenile onset spondylitis in which associated aortic, and possibly mitral, valve disease dominated the clinical course for many years before axial joint disease required total replacement of the right hip. PMID- 6288152 TI - GUEPAR hinge knee arthroplasty: a five-year followup. PMID- 6288154 TI - Selection of components for lower limb amputation prostheses. PMID- 6288153 TI - Correction of flexion deformity of the knee by supracondylar osteotomy. PMID- 6288156 TI - Bicycle-related accidents and injuries in a population of urban cyclists. AB - To gain a better understanding of urban bicycle-related accidents and injuries, 93 Manhattan cyclists were asked to fill out a questionnaire on the subject and the results of their responses were analyzed. They reported a total of 51 bicycle related accidents and 59 injuries of varying degrees of severity. Although 42 percent declared they used safety helmets, head trauma was reported by 25 percent of the injured cyclists. Most were not wearing protective headgear at the time of their injury, however. Increased cycling mileage and length of time cycling were significantly associated with increased accident occurrence. Surprisingly, 47 percent of the accident victims cited themselves as being at fault for the accident in which they were injured. PMID- 6288155 TI - The clinical importance of articulations of the fibula. AB - Abnormal articular surfaces of the tibiofibular joint may result in painful knees. Aberrations in the tibiofibular joint may be secondary to old fractures of the bones of the ankle or the bones of the ipsilateral leg. Similarly, neuromuscular disease with growth abnormalities or primary osteoarthritis of the tibiofemoral joint may initiate secondarily abnormalities in the joint in question. There is a definite close relationship between mobility of the bones of the ankle and the status of the upper tibiofibular joint. For that reason, this articulation should be included in the routine examination of the knee joint. PMID- 6288157 TI - Open fractures of the shaft of the tibia treated by intramedullary nailing. AB - The technique and results of treating 57 selected open tibial shaft fractures by intramedullary nailing are reviewed. All but three of the the fractures were located in the middle third of the tibia; two thirds of these were transverse or short obliques. Excellent or good results were obtained in 87.7 percent of all cases. The mean time to union of Type I fractures was 12.6 weeks, of Type II fractures 15 weeks, and of Type III fractures 20.5 weeks. Complications included two infections in two of the Type III fractures. PMID- 6288160 TI - Serologic survey of hepatitis A antibodies in Mexican children. PMID- 6288158 TI - Dietary fiber and disease. PMID- 6288159 TI - Artificial food colors and childhood behavior disorders. PMID- 6288161 TI - Ion recombination loss in ionization chambers irradiated by proton beams. PMID- 6288162 TI - Absence of splenic uptake of radiocolloid due to Thorotrast in a patient with Thorotrast-induced cholangiocarcinoma. PMID- 6288163 TI - Trace element analysis of urinary calculi using proton induced X-ray emission (PIXE). PMID- 6288164 TI - Ulnar nerve palsy and walking frames. PMID- 6288165 TI - High serum vitamin B12 binding capacity as a marker of the fibrolamellar variant of hepatocellular carcinoma. AB - Ten (9.3%) of 107 patients with hepatocellular carcinoma had considerably increased serum unsaturated vitamin B12 binding capacity. All 10 were young (mean 12 years), had no serum alpha-fetoprotein, and no underlying cirrhosis; all had a longer survival compared with patients without increased serum unsaturated vitamin B12 binding capacity in the study. Seven of the 10 patients had fibrolamellar hepatocellular carcinoma, a recently recognised histological variant, which was found in only one young patient without increased serum unsaturated vitamin B12 binding capacity and no alpha-fetoprotein among the remaining 97. This high degree of correlation between increased serum unsaturated vitamin B12 binding capacity and fibrolamellar hepatocellular carcinoma has not been reported before. Increased serum unsaturated vitamin B12 binding capacity may be of considerable help in diagnosis, prognosis, and monitoring treatment of this well-defined group of patients with hepatocellular carcinoma but no alpha fetoprotein. PMID- 6288166 TI - Fungal arthritis simulating juvenile rheumatoid arthritis. AB - Petriellidium boydii is often isolated from maduromycosis but has recently been associated with arthritis. A previously healthy 6-year-old boy developed chronic purulent arthritis of the knee after a bicycle accident. Culture of aspirate grew no pathogens and antibiotic treatment had no effect. Culture of synovial fluid grew P boydii, which responded initially to amphotericin but reappeared after six months. Subsequent treatment with miconazole was stopped after development of haematuria. The fungus was sensitive to ketoconazole, and treatment with this drug cured the infection. With the introduction of ketoconazole it is of practical importance to recognise fungal infections. PMID- 6288167 TI - Idiopathic hypoparathyroidism presenting as urinary and faecal incontinence. PMID- 6288168 TI - Regulation of acetylcholine synthesis in normal and neurotropic viral infected sympathetic ganglia. PMID- 6288169 TI - Ontogenesis of alpha- and beta-receptors located on cerebral microvessels. AB - Adrenergic receptors in rat brain microvessels were studied during ontogenesis. Microvessels were prepared by albumin floatation and glass bead filtration techniques from cortices of 10, 20- and 90-day-old rats. The lower level of alpha 1- alpha 2- and beta-receptor sites observed in early life may correlate with the lower capacity of cerebral vascular regulatory mechanism in this period. PMID- 6288170 TI - Regulation of glutamate receptors: possible role of phosphatidylserine. AB - The polar head group of the phospholipid phosphatidylserine is similar in structure to the glutamate analogue 2-amino-4-phosphonobutyric acid (APB), an antagonist of excitatory transmission in the brain. When tested in ligand binding assays phosphatidylserine and its polar head group components O-phosphoserine and L-alpha-glycerophosphoserine were good displacers of APB-sensitive L-glutamate binding. The polar head group components were also antagonists of synaptic field potentials in the rat dentate gyrus evoked by stimulation of a presumed glutamate using pathway. These results suggest that phosphatidylserine may regulate the activity of synaptic L-glutamate receptors. PMID- 6288171 TI - Two muscarinic depolarizing mechanisms in mammalian sympathetic neurons. AB - A voltage-sensitive outward membrane current ('M') and a consequent change in conductance (delta G) appear with a slow time-constant, in principal neurons of rabbit superior cervical ganglion (SCG), only when membrane potentials (Vm) are depolarized to less than -60 mV. Effects of muscarine on the voltage-current curves indicate that, in this depolarized range of less than -60 mV, suppression of M-current could contribute a muscarinic depolarization accompanied by a decrease in G; but that, at all Vms tested (about -90 to -40 mV), there is an additional larger muscarinic depolarization with no delta G. Thus, the muscarinic depolarizing response and the equivalent slow excitatory postsynaptic potential in the rabbit SCG may consist of two different components: one is due to the suppression of M-current and is substantial only in the depolarized range; the other is probably mediated via an intracellular increase in cyclic GMP and can account for most or all of the response at Vms more negative than -55 mV. PMID- 6288172 TI - Chronic vitamin D deficiency in the weanling rat alters catecholamine metabolism in the cortex. AB - The effect of a vitamin D deficient (--D), vitamin D replete (150 I.U. D3 twice weekly) and normal rat diets for 4 weeks in weanling male rats on the steady state concentration in several brain sites of dopamine (DA), norepinephrine (NE), and dihydroxyphenylacetic acid (DOPAC) was investigated. The areas of the central nervous system assayed were the brainstem, cerebellum, cerebral cortex, hypothalamus-median eminence, and striatum. The results indicate that DA content of cortex and hypothalamus significantly increased in the --D group compared to the normal diet or D replete groups. The concentrations of DOPAC and NE in the cortex of both --D and D replete rats increased significantly compared to normal diet group. Plasma calcium level was significantly lower in --D group compared to the normal diet or vitamin D replete groups. PMID- 6288173 TI - CCK-8-Immunoreactivity distribution in human brain: selective decrease in the substantia nigra from parkinsonian patients. AB - The regional distribution of immunoassayable CCK-8 was determined in 12 regions of control and parkinsonian human brain, with specific attention to the possible regional coexistence of CCK-8 with dopamine. In Parkinson's disease, CCK-8-I levels were only decreased in the substantia nigra where dopamine cell bodies lie, and not in striatal and corticolimbic dopamine projecting areas. Our results suggest that the major proportion of dopaminergic neurones degenerated in Parkinson's disease may not contain the CCK-8 peptide. PMID- 6288174 TI - Drug reinforcement studied by the use of place conditioning in rat. AB - Rats display a preference for an environment in which they previously received morphine. The present report provides behavioral and pharmacological data for this simple model of reinforcement produced by opiates and describes an aversion in rats for an environment in which they previously received naloxone. Preferences were produced with intravenous (i.v.) morphine sulfate at doses of 0.08-15 mg/kg and durations of the pairing between environment and morphine of 10 min to 1.5 h. Preferences were also seen with other opiate agonists (etorphine HCl and levorphanol-tartrate), another route of drug administration (subcutaneous), and after 1-4 administrations of morphine. Cocaine-HCl (i.v.), a non-narcotic drug, known to be self-administered by humans, also produced a place preference. Lithium chloride (i.v.), an agent found to be a punishing stimulus in other situations, produced a place aversion. There was no appreciable preference for an environment paired with dextrorphan-tartrate and naloxone-HCl (2 mg/kg, i.p.) blocked the production of the preference produced by i.v. morphine. In contrast to the effect produced by morphine, aversions were produced with (-) naloxone-HCl alone at doses of 0.1-45 mg/kg (i.v.). The aversion was not produced at (+)-naloxone. Implantation of rats with a 75 mg morphine pellet 3 days prior to place conditioning potentiated the aversive effect of naloxone. It was concluded that place conditioning produced by morphine and naloxone is mediated by specific opiate receptors and that stimulating and decreasing activity of the endogenous opioid peptide system with systemically administered drugs is positively reinforcing and aversive, respectively. The discussion emphasizes application of the simple and sensitive place conditioning model to drug reinforcement research, including analyses of reinforcement produced by microinjection of opiates into the brain. PMID- 6288175 TI - Control of calcium current in rat sympathetic neurons by norepinephrine. AB - Inward voltage-dependent calcium currents were recorded from clamped rat sympathetic ganglion cells using either one or two microelectrodes. Suppression of potassium current was achieved by applying tetraethylammonium (TEA) externally and TEA plus cesium internally. Peak ICa was observed at 0 mV. ICa was abolished by perfusing cadmium or low calcium medium. ICa was reduced by adding norepinephrine (1-50 micrometers). This effect was not accompanied by any major change in the voltage sensitivity or time course of the residual calcium current. It is suggested that norepinephrine acts by reducing the number of available calcium channels. PMID- 6288176 TI - GABA binding and bicuculline in spinal cord and cortical membranes from adult rat and from mouse neurons in cell culture. PMID- 6288177 TI - GABA and bicuculline actions on mouse spinal cord and cortical neurons in cell culture. AB - The neutral amino acid gamma-aminobutyric acid (GABA) produced membrane hyperpolarization and increased membrane chloride ion conductance of spinal cord (SC) and cortical (CTX) neurons in cell culture. GABA dose-response curves were obtained for SC neurons by pressure applying known concentrations of GABA from micropipettes with large tips (miniperfusion pipettes). GABA response threshold was about 2 micrometers and large responses were elicited at GABA concentrations greater than 10 micrometers. Bicuculline (BICUC) (0.1-10 micrometers) reversibly antagonized GABA responses on both SC and CTX neurons with a half maximal inhibitory concentration of about 1 micrometer. BICUC antagonism of GABA responses was competitive (Lineweaver-Burke analysis). These results are compared with data on GABA and BICUC displacement of [3H]GABA binding to membranes of SC and CTX neurons in cell culture. It is suggested that high affinity GABA receptors are likely to be relevant for postsynaptic GABA responses while low affinity GABA receptors may be presynaptic. PMID- 6288178 TI - Responses of mechanoreceptive afferent units in the glabrous skin of the human hand to sinusoidal skin displacements. AB - The impulse responses to perpendicular sinusoidal skin displacements were recorded from 4 different types of mechanoreceptive different units innervating the glabrous skin of the human hand. The cycle responses, defined as the number of impulses evoked per sine wave cycle, were studied at a wide range of frequencies (0.5-400 Hz) and amplitudes (0.001-1mm). The rapidly adapting units (RA) were most easily excited at stimulus frequencies between 8 and 64 Hz, whereas the corresponding frequencies for the Pacinian units (PC) were above 64 Hz. However, at high stimulus amplitudes, the RA and the PC units showed quite similar response profiles within the range of frequencies tested. The sensitivities of the slowly adapting unit types (SA I and SA II) were greatest at lower frequencies. A characteristic finding for all 4 types of units was that the higher the amplitude, the lower the frequency at which the cycle response was maximal. PMID- 6288179 TI - In vitro labeling receptor autoradiography: loss of label during ethanol dehydration and preparative procedures. AB - Slide-mounted tissue sections of brain were incubated with several reversibly binding [3H]ligands to label receptors. Exposure of these labeled, mounted tissue sections to ethanol solutions for dehydration resulted in a substantial loss of receptor bound ligands in all cases, even when the tissues were fixed with formaldehyde vapors before exposure. Thus, serious problems can be introduced into in vitro labeling autoradiographic procedures by exposure of sections to aqueous or organic media. PMID- 6288180 TI - Lack of specificity of a "taurine antagonist". PMID- 6288181 TI - Sensitivity to edges of mechanoreceptive afferent units innervating the glabrous skin of the human head. AB - One type of rapidly adapting (RA) and one type of slowly adapting (SA I) mechanoreceptive unit innervating the glabrous skin area of the human hand have small and well defined receptive fields. Their responses to perpendicular sinusoidal skin displacements were measured when the field was completely covered by a flat contact surface and when the edge of the contact surface was placed over the field. It was found that the majority of the RA and SA I units exhibited stronger responses when the edge of the contactor was placed over the receptive field. This edge sensitivity, which was greatest for the SA I units, may be of particular importance in improving the spatial analysis of the edges of objects in contact with the skin during manual manipulation and exploration. PMID- 6288182 TI - Plasticity of neostriatal dopamine receptors after nigrostriatal injury: relationship to recovery of sensorimotor functions and behavioral supersensitivity. AB - Rats given unilateral injections of 6-OHDA along the course of the mesotelencephalic dopaminergic projection show impairments in contralateral sensorimotor functions from which they often recover. Such rats also display an enhanced sensitivity to DA receptor stimulants, e.g. apomorphine, as revealed by contralateral turning, and an increased binding of neuroleptic compounds (e.g. [3H]spiroperidol) to the denervated striatum. This research examines the relationship of these receptor changes to both behavioral supersensitivity and recovery of sensorimotor functions by quantifying the time course of each phenomenon after injury. The supersensitivity to apomorphine and the behavioral recovery developed with a similar time course after injury, being evident within 1.5-3 days and reaching nearly maximal levels by 2 weeks postoperatively. A significant increase in in vivo [3H]spiroperidol binding to the denervated striatum occurred by 4 days postoperatively, and the magnitude of this change increased linearly during the first postoperative month. In contrast, the in vitro binding of this ligand to membranes of the denervated striatum was not increased until 3 weeks after the lesion. The results suggest that a proliferation of DA receptors may contribute to the pharmacological supersensitivity and the recovery of function, and that these early receptor changes may be revealed with greater sensitivity using in vivo binding techniques. PMID- 6288183 TI - Cholinergic mechanisms and neurotransmission in the cerebellum of the rat. An in vitro study. PMID- 6288185 TI - Regional distribution of cholinergic neurons in human spinal cord transections in the patients with and without motor neuron disease. AB - Regional distribution of enzymic activities in acetylcholine (ACh) metabolism was examined on thinly-sectioned transverse slices of human spinal cords obtained during autopsy of 5 motor neuron disease (MND) and 5 control patients without MND. Choline acetyltransferase (ChAT) activity was highly concentrated in the ventral horn regions (gray and white matters) of cervical, thoracic and lumbar spinal cord of non-MND patients. This enzyme activity was found to be remarkably low in the ventral gray and white matter of MND patients compared with that of the controls. Although the distribution of acetylcholinesterase (AChE) activity was found to be high in both ventral and dorsal gray matter of the spinal cord, little difference was observed between each corresponding region of MND and control patients, except relatively low enzyme activity in the cervical ventral horn region of MND patients. Muscarinic cholinergic receptors, examined as specific [3H]quinuclidinylbenzilate ([3H]QNB) binding, was also highly concentrated in the ventral and dorsal gray matter of the control spinal cord, and was strongly reduced in the ventral horn region of MND patients, indicating a quite similar distribution pattern of ChAT activity. These biochemical changes of cholinergic transmission system may be paralleled to the morphological degeneration of the spinal lower motor neurons in MND patients. Activity of 2',3' cyclic nucleotide-3'-phosphodiesterase (CNPase), a marker enzyme of central myelin structure, was evenly distributed throughout the whole spinal cord section, without regard to the gray and white matter, of both MND and control patients. PMID- 6288184 TI - Possible involvement of serotonergic neurons in the reduction of locomotor hyperactivity caused by amphetamine in neonatal rats depleted of brain dopamine. AB - This experiment attempted to determine the mechanism by which amphetamine reduces locomotor hyperactivity in neonatal rats given brain dopamine (DA)-depleting 6 hydroxydopamine (6-OHDA) injections. Brain DA neurons were destroyed selectively in neonatal rats by intraventricular (i.v.t.) injections of 6-OHDA following desmethylimipramine (DMI) pretreatment. Control rats received DMI and i.v.t. injections of the 6-OHDA vehicle solution. Rats given the 6-OHDA treatment displayed 7-fold increases in locomotor activity compared to controls during days 16-55 of life. Throughout this period, amphetamine (1 mg/kg) reduced locomotor hyperactivity in 6-OHDA-treated rats but increased locomotor activity in control rats. The reduction of hyperactivity caused by amphetamine (0.5-4 mg/kg) was dose related and was not accompanied by stereotyped behavior. Like amphetamine, methylphenidate (4 mg/kg) reduced locomotor hyperactivity in rats given 6-OHDA. The DA antagonist, spiroperidol (50-200 micrograms/kg) failed to attenuate the hyperactivity-reducing effect of amphetamine in 6-OHDA-treated rats at doses which abolished the stimulant effect of amphetamine in control rats. However, the serotonin antagonist methysergide (0.5-4 mg/kg) produced dose-dependent antagonism of the effect of amphetamine in 6-OHDA-treated rats. Pretreatment with propranolol ((5 mg/kg), phentolamine (5 mg/kg), atropine (0.5 mg/kg) or naloxone (10 mg/kg) failed to alter the reduction in locomotor hyperactivity caused by amphetamine. The serotonin releasing agent, fenfluramine (3 mg/kg), and the serotonin agonist, quipazine (0.5-4 mg/kg), both reduced locomotor hyperactivity in 6-OHDA-treated rats while not altering locomotion in control rats. These results confirm previous observations that amphetamine reduces locomotor hyperactivity caused by neonatal 6-OHDA administration and suggest that this effect is mediated by increased serotonergic neurotransmission. PMID- 6288187 TI - Long-lasting cardiovascular depressor response following sciatic stimulation in spontaneously hypertensive rats. Evidence for the involvement of central endorphin and serotonin systems. AB - A naloxone-reversible long-lasting depressor response induced by a prolonged low frequency stimulation of the sciatic nerve in conscious spontaneously hypertensive rats (SHRs) was reported in a previous paper. In the present study pharmacological tools were used to further investigate the neurotransmitters involved in this phenomenon. Naloxone infusion (20--25 mg/kg/h following a bolus dose of 10 mg/kg i.v.) attenuated significantly the depressor response, while dexamethasone pretreatment had no such effect, suggesting an important role of the brain endorphin system, but not of the pituitary beta-endorphin, in this depressor response. Since the concomitant increase in pain threshold produced by the sciatic stimulation exhibited a different time course of development and naloxone reversibility, it is suggested that the depressor response and the hypalgesic effect produced by the same stimulation are mediated via different types of opiate receptors in the brain. On the other hand, PCPA abolished the post-stimulatory depressor response whereas 5-HTP and zimelidine had additive effects on the sciatic stimulation-induced depressor response, suggesting the involvement of central serotonin systems in the mechanism of the response. The interaction between the central endorphin and the serotonin systems in the mediation of the post-stimulatory depressor response is discussed. PMID- 6288186 TI - Uptake of horseradish peroxidase at isolated nerve terminals in relationship to transmitter release; biochemical results. AB - The uptake of horseradish peroxidase (HRP) into isolated nerve terminals (synaptosomes) has been studied by using a spectrophotometric method to determine the enzyme activity. HRP is rapidly taken up by synaptosomes, it is not removed by multiple washes in iso-osmotic medium but is lost if the particles are ruptured by hypo-osmotic conditions. The uptake is not affected by metabolic poisons, is reduced at lower temperatures and is not associated with any significant release of cytoplasmic lactate dehydrogenase suggesting an endocytotic mechanism. Intra-synaptosomal HRP can be released by a process that is similar to uptake and is also not accompanied by any loss of synaptosomal lactate dehydrogenase indicating exocytosis. Depolarization of synaptosomes (by high potassium concentrations) was found to release [14C]ACh but to have no effect on HRP uptake either simultaneously or after recovery in a non depolarizing medium. Absence of Ca2+ prevented depolarization evoked release of [14C]ACh but had no effect on the uptake of HRP. The release of HRP was not increased by depolarization even though [14C]choline taken up during the same period was released as [14C]ACh. It is concluded that the endo-exocytotic cycle that transports HRP across the synaptosomal membrane is unrelated to transmitter release. A discrete vesicular localization of HRP reaction product was only occasionally seen in the EM nor could consistent differences resulting from depolarization be observed. However, the ultrastructural localization was found to be unreliable because glutaraldehyde fixation irreversibly inactivated 80--90% of the HRP even when it was sequestered within synaptosomes and the insoluble reaction product precipitated from a supersaturated solution onto membranes. PMID- 6288188 TI - Studies on the role of calcium in adaptation of the crustacean stretch receptor. Effects of intracellular injection of calcium, EGTA and TEA. AB - The effects of intracellular pressure injection of Ca2+, EGTA and TEA on the receptor potential of the crayfish stretch receptor were studied. Injection of Ca2+ caused both the transient phase and the static phase of the receptor response to diminish in amplitude, the decrease being greater for the static phase. This phase was almost abolished after a few minutes of injection. Injection of EGTA caused a decrease in the amplitude of the transient phase and an increase of the static phase. These changes progressed during the injection and finally the receptor potential became almost square. After injection of TEA the static phase increased and approached the height of the transient phase making the response almost square. The results provide evidence for the important role of intracellular Ca2+ for the adaptation of the receptor. It is suggested that the adaptive decline of the receptor potential is due to an outward potassium current which is controlled by the intracellular concentration of Ca2+. PMID- 6288189 TI - Effect of age on beta-adrenergic receptors on cerebral microvessels. AB - The responsiveness of beta-adrenergic receptors in cerebral microvessels was studied in aged rats by measuring cAMP formation induced by norepinephrine and [125I]iodohydroxybenzylpindolol binding. The density of beta-receptor sites is reduced by aging in the rat cerebral microvessels. These results suggest that the changes in brain circulation during aging may be at least partially due to a reduction of the beta-adrenergic control of cerebral microvessels. PMID- 6288190 TI - Black widow spider venom: excitatory action on hippocampal neurons. AB - Electrical activities of thin hippocampal sections of the guinea pig were studied in vitro during and after administration of black widow spider venom (BSV). In CA3 neurons, BSV induced spontaneous discharges which were suppressed by Co2+ or Mn2+. During BSV administration, trains of potential changes resembling excitatory postsynaptic potentials (EPSPs) were recorded intracellularly. The EPSP-like potentials were blocked by TTX. BSV seems to induce impulses in presynaptic fibers or terminals and thereby facilitates release of neurotransmitters. PMID- 6288191 TI - The effect of naloxone on normal human sleep. AB - Placebo and naloxone (4, 8, and 12 mg) were infused on separate nights in a double-blind fashion over a 20-min period between the 25th and 45th min of sleep in normal volunteers. Naloxone produced a dose-dependent increase in REM latency (the time from onset of sleep until the first REM period), and duration of the first REM period and second NREM period. The number of REM periods was also reduced. PMID- 6288193 TI - Phencyclidine blockade of sodium and potassium channels in neuroblastoma cells. PMID- 6288192 TI - Cyclic nucleotide modulation of Na+ and K+ currents in the isolated node of Ranvier. PMID- 6288194 TI - DL-quisqualic and L-aspartic acids activate separate excitatory conductances in cultured spinal cord neurons. PMID- 6288195 TI - Dopaminergic terminal excitability following arrival of the nerve impulse: the influence of amphetamine and haloperidol. AB - Variations in the excitability of the axons and terminal fields of dopaminergic neurons of the substantia nigra were examined as a function of time following the nerve impulse in urethane-anesthetized, immobilized rats. Excitability was measured by determining the threshold, defined as the minimum current required to consistently activate dopaminergic neurons antidromically. Threshold currents were maximal immediately following the action potential and declined exponentially to a plateau. The interval during which threshold current declined, the phasic period, was significantly longer for stimulation of neostriatal terminal fields as contrasted to stimulation of axons along the medial forebrain bundle. A positive correlation was observed between antidromic response latency and the duration of this phasic period for both sites of stimulation, an observation consistent with the view that the site of initiation of the antidromic action potential is of smaller diameter in the neostriatum than in the medial forebrain bundle Amphetamine, which promotes dopamine release and/or re uptake blockade, reduced dopaminergic terminal excitability in the neostriatum at all intervals examined. This effect increased at successively shorter intervals during the phasic portion of the excitability curve. Haloperidol, a dopamine antagonist, increased the excitability of dopaminergic terminal fields, an effect which was also more marked earlier in the phasic interval. Neither amphetamine nor haloperidol had a significant effect on the excitability of dopaminergic axons in the medial forebrain bundle. Variations in dopaminergic terminal excitability after impulse arrival, and the effects of amphetamine and haloperidol on this phenomenon suggest that terminal excitability is determined by events related to arrival of the nerve impulse including activation of presynaptic 'autoreceptors' by dopamine released from the synaptic ending. PMID- 6288196 TI - Membrane phosphoproteins of rat hippocampus: sensitivity to tetanic stimulation and enkephalin. AB - Hippocampal slices are electrically stimulated in the perforant path with a pulse train, which can lead to long-term potentiation (LTP). Of the thus stimulated slices, subcellular fractions are prepared and used in an endogenous protein phosphorylation assay. A phosphoprotein band which was reported earlier to be sensitive to electric stimulation as well as to methionine-enkephalin is now further analyzed: it consists of two phosphoproteins only slightly differing in molecular weight: 50,000 Mr (50 K) and 52,000 Mr (52 K), but having distinct biochemical properties and subcellular localization. Their IEP is dissimilar (3.5 4.3 and 5.3, respectively), they display different sensitivity towards calcium when tested in the phosphorylation assay, but are both cAMP-independently phosphorylated. Only one of them responds to tetanic stimulation with an increased phosphorylation post hoc. This protein, the 52 K component, is localized in synaptic membranes. Moreover, this protein also responds to incubation of slices with methionine-enkephalin. The phosphorylation of the 50 K component is not influenced by electric stimulation, nor by incubations with neuropeptides; its phosphorylation takes place in material sedimenting with the mitochondrial cell fractions and is strongly calcium- and calmodulin-dependent. PMID- 6288197 TI - A 14C-2-deoxyglucose analysis of the neural pathways of the limbic forebrain in the rat: II. The hypothalamus. AB - An attempt was made to characterize the nature of the functional organization of the hypothalamus by observing the patterns of uptake of 14C-2-deoxyglucose (2DG) following electrical stimulation of different regions within the preoptico hypothalamus in the rat. The experimental paradigm consisted of electrical brain stimulation delivered continuously for periods of 30 sec on and 30 sec off for 45 minutes following injection of 2DG. Brains were removed and processed for autoradiography. Activation of the medial forebrain bundle was noted following stimulation of the nucleus accumbens and lateral preoptico-hypothalamus. Activated fibers could be followed only in a caudal direction through the medial forebrain bundle and into the ventral tegmental area as a result of nucleus accumbens stimulation. Stimulation of the lateral preoptic region or of the anterior half of lateral hypothalamus produced activation of the lateral septal nucleus, lateral habenular nucleus, perifornical region, midline thalamus and ventral tegmental area. Since stimulation of the perifornical hypothalamus significantly activated the rostro-caudal extent of the midbrain cental gray, it is suggested that impulses from the lateral hypothalamus reach the lower brainstem via its connections with the perifornical hypothalamus. Ventromedial hypothalamic stimulation activated only the lateral septal nucleus, cortico medial amygdala and medial preoptico-hypothalamus, while medial preoptico hypothalamic stimulation resulted in increased 2DG uptake in the midbrain central gray, thus suggesting that medial hypothalamic impulses reach the brainstem by first ascending to the level of the preoptico-hypothalamus. Mammillary body stimulation orthodromically activated fibers in the mammillothalamic and mammillotegmental tracts and antidromically fibers in the fornix for a short distance. PMID- 6288198 TI - High frequency discharges in sensory nerves following ischemia. AB - The effects of ischemia and recovery from ischemia on afferent discharges recorded from dorsal root filaments were assessed in anesthesized cats. During ischemia produced by cross-clamping the lumbar aorta, some of the hindlimb afferents originating in muscle spindles exhibited a sustained high frequency discharge of 120--160 Hz, the so-called "explosion." During recovery from ischemia, nerve discharges appeared in previously inactive dorsal root afferent nerve fibers not of muscle spindle origin. The post-ischemic high frequency discharges appeared in bursts of action potentials at frequencies of 200--500 per second, with 6--32 spikes per burst; the bursts recurred at intervals of 120--600 msec. The nerve fibers exhibiting post-ischemic bursts could not be activated by muscle stretch, muscle tension, palpation, or tactile skin stimulation prior to or during the ischemia. No post-ischemic burst discharges could be detected in analogous experiments with the sural nerve. Intermediate size fibers from normal cutaneous and muscle mechanoreceptors were eliminated as possible origins of the post-ischemia activity. Possible sources include dorsal root ganglion cells or fibers whose sensory endings lie in deep structures such as the walls of the larger blood vessels. PMID- 6288199 TI - beta-Adrenergic, but not benzodiazepine, binding sites are reduced in dystrophic mouse brain. AB - We have examined brain tissue from dystrophic mice (male 129/ReJ-dy and female 129 B6F1/J-dy) to determine whether CNS lesions may accompany the known muscular defects. In all brain regions examined we found a significant reduction in the numbers of beta-adrenergic binding sites when compared to brain from control mice. In contrast, the number of benzodiazepine binding sites appeared normal. These changes in adrenergic sites may be related to the known abnormally high excretion of catecholamines. Our results also emphasise the need to study extra muscular sites in this disease. PMID- 6288200 TI - Neurofibrosarcoma and irradiation. PMID- 6288201 TI - Wilms' tumor and neurofibromatosis. PMID- 6288202 TI - Sulphide as an inhibitor and electron donor for the cytochrome c oxidase system. AB - Anomalies both kinetic and equilibrium in nature are described for the inhibition of cytochrome c oxidase activity by sulphide in the isolated enzyme and in submitochondrial particles. These anomalies are related to the involvement of more than 1 mol of sulphide in the blockage of one cytochrome aa3 centre. Sulphide reduces resting cytochrome a3, a reaction that results in oxygen uptake and the loss of a sulphide molecule. Sulphide can also reduce cytochromes c and a; in the former case, a part of the one-equivalent oxidation product, presumed to be the SH radical, reacts with oxygen. Such oxygen uptake is also seen under aerobic conditions when ferricyanide reacts with sulphide. Three phases are identified in the inhibitory interaction of sulphide with the cytochrome c oxidase enzyme itself: an initial rapid reaction involving sulphide oxidation, oxygen uptake, and conversion of cytochrome aa3 into the low-spin "oxyferri" form; a subsequent step in which sulphide reduces cytochrome a; and the final inhibitory step in which a third molecule of sulphide binds the a3 iron centre in the cytochrome a2+ a3 3+ (oxy) species to give cytochrome a2+ a3 3+ H2S. the initial events parallel some of the events in the interaction of the cytochrome c cytochrome aa3 system with monothiols; the final inhibitory event resembles that with cyanide. PMID- 6288203 TI - A topoisomerase from chicken erythrocyte nuclei which does not assemble nucleosome core particles in vitro. AB - We have examined the ability of a topoisomerase purified from chicken erythrocyte nuclei to mediate nucleosome core assembly in vitro at physiological ionic strength (0.15 M NaCl). Although we have detected limited amounts of spontaneously assembled nucleosome cores at this salt concentration, the addition of this topoisomerase does not increase the amount of assembly observed. Nucleosome assembly was assayed by quantitating the amount of core particle length DNA accumulated with time upon the nuclease digestion of histone-DNA complexes. In addition, the amount of negative supercoils introduced into relaxed closed circular DNA upon nucleosome core particle assembly was determined. Correctly assembled complexes do not protect more DNA from nuclease digestion than random histone-DNA complexes but shift the heterogeneous size distribution of protected fragments to a more homogeneous distribution centred around 145 base pairs. Under our conditions of nucleosome assembly, a second histone-DNA complex which is distinct from the nucleosome core can be detected under physiological ionic strength conditions. This particle does not form in high salt assembly experiments. Similarly, the assembly of this particle is unaffected by the presence or absence of topoisomerase. PMID- 6288204 TI - Biosynthesis of CDP-diacylglycerol in hog mesenteric lymph node lymphocytes. AB - The synthesis of CDP-diacylglycerol by CTP: phosphatidate cytidylyltransferase (EC 2.7.7.41) has been studied in microsomes isolated from hog mesenteric lymph node lymphocytes. The properties of this enzyme were found to be similar in many respects to that described in rat liver microsomes; however, it is not stimulated by GTP or any other nucleotides under normal assay conditions. The enzyme requires that the phosphatidate be emulsified in a cationic detergent for optimal activity. The quaternary ammonium phospholipids lecithin and sphingomyelin were found to stimulate the formation of CDP-diacylglycerol even in the presence of optimal cationic detergent. Other phospholipids or detergents had no effect or were inhibitory to the reaction. Only in the presence of either lecithin or sphingomyelin did nucleotides such as ATP, GTP, UTP, and ITP stimulate the formation of CDP-diacylglycerol. The Km and Vmax for CTP were found to be 0.6 and 1.2 mM, respectively, while the apparent Km and Vmax for phosphatidate were 0.65 and 1.2 mM. Magnesium was found to be the only metal ion that stimulated the reaction, with an optimal concentration of 20 mM. Fluoride ions at 20 mM inhibited the reaction to the extent of 70%. The enzyme was found to be very unstable when the isolated microsomes were stored at -20 degrees C for 24 h, losing approximately 75% of its activity. PMID- 6288205 TI - A microassay for UDP-N-acetylglucosamine pyrophosphorylase. PMID- 6288206 TI - Cholinephosphotransferase activities in microsomes and neuronal nuclei isolated from immature rabbit cerebral cortex: the use of endogenously generated diacylglycerols as substrate. AB - A neuronal nuclear fraction (N1) and a microsomal fraction (P3) were isolated from homogenates of cerebral cortices of 15-day-old rabbits. A nuclear envelope fraction (E) was prepared from N1. To assay cholinephosphotransferase, diacylglycerols were first generated in the membranes of these subfractions using a phospholipase C (Bacillus cereus) preincubation. With levels of endogenous diacylglycerols producing maximal specific cholinephosphotransferase activities, an activity ratio of 1:1:5 was found for N1, P3, and E, respectively. An independent neuronal nuclear cholinephosphotransferase, concentrated in nuclear membranes, is indicated. With regard to changes in pH and concentrations of MgCl2 and CDP-choline, N1 and P3 activities responded in a similar manner. However, in contrast to P3, N1 activities we much more profoundly inhibited at low levels of Triton X-100 (0.01-0.02 w/v%) and N1 showed quite significant levels of cholinephosphotransferase activity in the absence of a phospholipase C preincubation. Choline phosphotransferase in N1 and P3 showed Km values for CDP choline (0.028 and 0.031 mM, respectively) which were much lower than corresponding literature values determined using exogenous diacylglycerols as substrates for this enzyme. The presence of cholinephosphotransferase in neuronal nuclear membranes reflects a rather exceptional nuclear autonomy. This may be related to a need to maintain nuclear phospholipid in the absence of a well developed endoplasmic reticulum at early stages of neuronal development or to synthesize phospholipid in response to functions unique to the nucleus. PMID- 6288207 TI - The separation and properties of phosphoprotein phosphatase modulators from brown adipose tissue of young rats and the changes in their concentration during development. AB - The heat-stable modulators of phosphoprotein phosphatase activity have been partically purified from brown adipose tissue. A nonphosphorylatable inhibitor of phosphorylase phosphatase (inhibitor 2) and an activator of phosphohistone phosphatase were similar to the corresponding modulators from muscle and liver in both their physical properties and in their relative effects upon three different preparations of phosphatase. An inhibitor of phosphorylase phosphatase that was only active when phosphorylated was eluted from DEAE-cellulose by 80 mM NaCl (inhibitor 1'). Only a small amount of inhibitor was eluted at 20 mM NaCl (inhibitor 1), which is the concentration that eluted the bulk of the phosphorylatable inhibitor in muscle and liver. Inhibitor 1 and inhibitor 1' had similar physical properties but differed in their activities towards the different phosphatases. It is suggested that they are different forms of the same protein and that inhibitor 1' more closely resembles the native inhibitor. The modulators did not compete with each other and were probably not subunits of a phosphatase complex. However, the direction and timing of the changes in their concentration in brown fat during the developmental period indicate that the inhibitors, at least, perform some useful physiological function in the tissue. The concentration of inhibitor 2 was high before birth and for 10 days after birth, when the tissue was proliferating. The concentration of the phosphorylatable inhibitor was highest immediately after birth and for the next 16 days, which is the period of greatest thermogenic activity of brown fat. PMID- 6288208 TI - Some properties of a mitochondrial endonuclease from yeast. AB - A mitochondrial endonuclease from Saccharomyces cerevisiae was previously shown to cut both strands of native DNA at opposite or nearby sites. The present studied demonstrate that the endonuclease activity is dependent on the strength of the hydrogen bonds between the DNA strands; the activity was measured at different ionic strengths, with substrates of different base compositions and also with DNA in which the double helix has been locally destabilized by ultraviolet irradiation, by depurination, and by single-stranded nicks. The activity is 30% greater with mitochondrial DNA (mt-DNA) than with nuclear DNA. At 0.08 ionic strength, the relative activities with double-stranded polydeoxyribonucleotides are 2.4:1:0.6 for poly(dA).poly(dU) : poly(dA).poly(dT) : poly(dG). poly(dC). Increasing ionic strength decreases similarly the activity with poly(dA).poly(dU) and poly(dA).poly(dT), but has little effect with poly(dG).poly(dC). The greater activity with poly(dA).poly(dU) than with poly(dA).poly(dT) was confirmed with nick-translated mt-DNA and with DNA synthesized in isolated mitochondria using [3H]TTP and [3H]dUTP in both cases. The endonuclease cuts modified DNA preferentially in the thymine dimer regions, at the apurinic sites, and at sites opposite to nicks. The possible involvement of this endonuclease in the degradation of mitochondrial DNA during "petite" induction is discussed. PMID- 6288209 TI - Effect of citrate on the reactions of cytochrome c with reductants and cyanide. AB - The rate of reduction of cytochrome c by ascorbate is decreased in the presence of anions. This decrease is due to two factors: (a) nonspecific changes in ionic strength which occur when the total ion concentration or the charge on the anion is altered and (b) "specific" binding of anions to a site on cytochrome c which directly inhibits the reaction. The reaction between cyanide ion and cytochrome c is also affected by anions: increasing the ionic strength decreases the apparent association rate constant for cyanide binding. Substitution of citrate for morpholinopropane sulphonate in isoionic buffer media increases the apparent dissociation constant for cyanide suggesting citrate stabilizes the cytochrome c heme crevice. Binding of cytochrome c to cytochrome aa3 also affects the Kd for the cyanide - cytochrome c complex indicating that cytochrome c bound to cytochrome aa3 does not react with cyanide as readily as does free cytochrome c. PMID- 6288210 TI - Improved carbohydrate tolerance in fibre-fed rats: studies of the chronic effect. AB - The effect of chronic consumption of diets containing either different sources of dietary fibre (8% guar, 8% pectin, or 8% multifibre) or low carbohydrate upon carbohydrate tolerance was examined in rats. Weight gain was significantly lower throughout the entire 28-day study period with the guar group and after 20 days with the multifibre group. When tested with a liquid meal (1 g sucrose/kg body weight) after 14 days on the diets, only the guar rats had significantly lower fasting and postprandial plasma glucose concentrations. After 28 days, the improved carbohydrate tolerance persisted in the guar rats and started to appear in the multifibre rats. Pectin and low carbohydrate diets had no effect upon either weight gain or carbohydrate tolerance. Consuming the fibre diets did not affect jejunal sucrase activities. Jejunal glucose uptake activity was significantly diminished when measured in fasting guar rats while postprandially activities were similar to controls. Jejunal Na-K-ATPase activities in fasting and postprandial guar rats were not related to changes in glucose uptake. These studies confirm that only certain types of dietary fibre improve carbohydrate tolerance and suggest that reduced weight gain and altered intestinal glucose uptake are factors involved in the chronic fibre effect. PMID- 6288211 TI - Absence of blocking effects on cardiac slow calcium channels by the intracellular calcium antagonist 2-n-propyl-3-dimethylamino-5,6-methylenedioxyindene. AB - Extensive pharmacological evidence supports the contention that 2-n-propyl-3 dimethylamino-5,6-methylenedioxyindene hydrochloride (pr-MDI) is a calcium antagonist with a predominantly intracellular site of action. On the other hand, electro-physiological evidence points to a possible membrane slow inward calcium channel blocking property of this agent. To gain further insight as to the site of action of pr-MDI, the interactions between the negative inotropic action of this agent and the positive inotropic actions of excess extracellular calcium (which directly penetrates the myocardial cells through the slow calcium channels), isoproterenol (which indirectly augments calcium influx through the slow calcium channels), and ouabain (which enhances calcium influx through membrane calcium entry routes distinct from the slow calcium channels) were investigated in the isolated, electrically drive guinea pig left atrium. Although excess extracellular calcium, isoproterenol, and ouabain reversed the negative inotropic effect of pr-MDI, an analysis of the concentration-response relationships to all three positive inotropic agents in the presence and the absence of pr-MDI demonstrated that this agent did not significantly inhibit the contractile effects of calcium, isoproterenol, or ouabain, at pr-MDI concentrations which exhibit intrinsic negative inotropic effects. It is concluded that pr-MDI does not block the membrane slow inward calcium channel nor other presumptive membrane routes of calcium entry into myocardial cells at concentrations of 10(-4) M or less. At very high concentrations (3 X 10(-4) M) some inhibition of slow channel calcium influx may occur. PMID- 6288212 TI - A simple device for growing Clostridium perfringens and its application in bacteriocin studies. AB - A simple device constructed of common laboratory material served as a minifermenter for the growth of Clostridium perfringens. A constant flow of nitrogen gas into a culture tube containing C. perfringens assured agitation of the culture and a mechanism for dispensing small volumes of liquid from the culture without disturbing the growth environment. The method was applied to examining the growth-inhibiting effect of bacteriocins of C. perfringens where a very economical use of radioactive isotopes was possible. The activity of some bacteriocins differed when compared with previous data obtained with stationary cultures. Two major categories of bacteriocin appear to exist for this species: those bacteriocins which block the incorporation of DNA, RNA, and protein precursors and those which interfere with the organism's cell wall. PMID- 6288213 TI - Radionuclide bone scan, radiographic bone survey, and alkaline phosphatase: studies of limited value in asymptomatic patients with ovarian carcinoma. AB - Bone scans or skeletal surveys were obtained in 104 patients with ovarian carcinoma. No metastases were identified at staging in the 43 patients with Stage I or II disease. Four patients in the entire series had osseous metastases. Three of the 40 patients with Stage III epithelial ovarian carcinoma had osseous metastases at the time of staging. All of these were Grade III lesions. One Stage I, Grade III patient demonstrated osseous metastases two years after initial diagnosis. None of the four patients with osseous metastases had an elevated alkaline phosphatase; three of the four had bone pain. Based on these results, it is suggested that radiographic bone survey and radionuclide bone scans are not indicated as screening procedures in asymptomatic patients with ovarian carcinoma. PMID- 6288214 TI - A specific antigenic defect of the basement membrane is found in basal cell carcinoma but not in other epidermal tumors. AB - The basement membrane of basal cell carcinoma was characterized by indirect immunofluorescence using antibodies to laminin, type IV collagen, and bullous pemphigoid antigen, three distinct protein components of basement membrane. Aggregates of basal cell carcinoma in the dermis were surrounded by a continuous basement membrane containing laminin and type IV collagen; however, bullous pemphigoid antigen was either completely undetectable or faint and discontinuous rather than linear. In contrast to this specific defect in bullous pemphigoid antigen found in basal cell carcinoma, well-differentiated superficially invasive epidermal squamous cell carcinoma and several benign epidermal tumors (trichoepithelioma, wart, keratocanthoma, seborrheic keratosis, cylindroma) displayed all three antigens in the basement membrane that surrounded epithelial cell aggregates. Cylindroma displayed in addition, broad areas of type IV collagen in the hyalinized material between epithelial islands. Basal cell carcinoma was the only tumor examined in which there was a specific antigenic defect in the basement membrane. This defect in bullous pemphigoid antigen may be due to abnormal synthesis by the tumor cells and could be related to the absence of differentiation of these cells. PMID- 6288216 TI - Lymphomatoid granulomatosis and Epstein-Barr virus. AB - A case report of a patient with lymphomatoid granulomatosis presenting initially as a reactivated Epstein-Barr virus infection is presented. Epstein-Barr virus is proposed in the possible role of establishing of an immunologically compromised state that may have set the stage for dissemination of this disease process. Of interest is the fact that successful chemotherapeutic management of the disease was accomplished using prednisone and cyclophosphamide. Furthermore, this clinical success was reflected in a decreasing Epstein-Barr virus early antigen antibody titers accompanied by increasing antivirus capsid antigen titers; hence, it appears that laboratory markers of the response of lymphomatoid granulomatosis to treatment are available in the form of soluble immune complexes, antibodies to Epstein-Barr virus-coded antigens early antigens, Epstein-Barr nuclear antigens and/or virus capsid antigens as well as the active E rosette assay for T-cells. Finally, these data, although supporting the role of Epstein-Barr virus in the pathogenesis of lymphomatoid granulomatosis, suggest the need for further study in additional patients to verify the results. PMID- 6288215 TI - Peripheral nerve damage during multiple myeloma and Waldenstrom's macroglobulinemia: an ultrastructural and immunopathologic study. AB - Peripheral nerve biopsies of 22 patients who were seen with a peripheral neuropathy were studied. On each occasion an ultrastructural study was performed and on 12 occasions an immunopathologic study was done. Ten patients had Waldenstrom's macroglobulinemia and 12 had multiple myeloma. Cellular infiltrates were observed in three cases of Waldenstrom's macroglobulinemia by light microscopy. A widening of the myelin sheath, corresponding to an accumulation of macroglobulin, was found in three cases of Waldenstrom's macroglobulinemia. The presence of abundant deposits of amyloid was noted in one case of multiple myeloma. These diverse features show the various possible mechanisms that can produce peripheral nerve damage during the course of malignant dysglobulinemias. The segmental demyelination is probably of immunologic origin. The axonal lesions cannot be explained satisfactorily except in those rare cases where amyloid deposits are present in the endoneurium. PMID- 6288217 TI - Lymphocytosis in a patient with malignant fibrous histiocytoma. AB - Malignant fibrous histiocytoma is an uncommon soft-tissue sarcoma. Patients with malignant fibrous histiocytoma may present with unusual clinical findings and an apparent association with malignant hematopoietic disease has been reported. We describe a patient in whom a diagnosis of chronic lymphocytic leukemia was made simultaneously with discovery of a large, retroperitoneal malignant fibrous histiocytoma. Following surgical debulking of the tumor, the patient's blood and bone marrow findings returned to normal. PMID- 6288218 TI - Histopathologic studies of undifferentiated carcinoma of the parotid gland. AB - Five-hundred-fifty-five primary epithelial tumors of parotid gland origin were examined. Eighteen cases could be defined undifferentiated carcinomas, constituting 3.2% of the total number or 10.2% of 176 malignant tumors. The undifferentiated carcinomas could be divided on the basis of the cell size into two types: (1) Small-cell type (12/18; 66.7%); the tumor cells were as large as, or slightly larger than lymphocytes. The cytoplasm was scanty with dark chromatin. Mitotic figures were numerous. The tumor cells in the periphery of the cell clusters exhibited a palisading arrangement. Electron microscopy revealed that these cells were rich in tonofilaments and connected by desmosomes. (2) Large-cell type (6/18; 33.3%); the carcinoma cells were roughly twice as large as those of the small-cell carcinoma. The tumor cells were spheroidal or spindle in shape with rich and clear cytoplasm. Many of the tumor-cell clusters showed macroalveolar structures, suggesting the possibility that this type occurred as a variant of poorly differentiated adenocarcinoma or epidermoid carcinoma. Clinicopathologically, there was no sex difference in the total number of the undifferentiated carcinomas, but the large-cell carcinomas were frequently encountered in females (83.3%). As to the age distribution, the carcinomas were found frequently in middle-aged patients from 30-49 years (66.7%). Many of the tumors were 2-4.9 cm at their greatest diameter (55.6%). Tumor mass less than 1.9 cm could not be found. PMID- 6288219 TI - Glomus tumor: a clinicopathologic and electron microscopic study. AB - This clinicopathologic study concerns 63 instances of glomus tumor of the soft tissues. The tumors occurred at different ages but were more common in earlier adult life and most commonly occurred on the fingers (35 cases), frequently as subungual nodules (26 cases). Other sites of occurrence were the forearm in seven, the knee in seven, and the leg in six. All patients but one complained of pain. Histologically, the tumors, showing endothelium-lined vascular spaces surrounded by masses of epithelioid cells, could be divided into three varieties: vascular (29 cases), myxoid (23 cases), and solid (11 cases). Under electron microscope the tumor cells in four cases proved to be smooth-muscle cells. The clinicopathologic evidence presented supports the hypothesis that the glomus tumor is a tumor-like lesion of mesodermal disorder rather than a true neoplasm. PMID- 6288220 TI - The growing teratoma syndrome. AB - Six patients with metastatic mixed germ-cell tumors who had been treated successfully with chemotherapy had recurring solitary enlarging masses. Four had enlarging pulmonary masses and two patients had enlarging abdominal masses. Each had the presumed chemotherapy refractory mass surgically resected and was found to have mature teratoma with absence of malignant histologies. The growth in two patients can be attributed to tense and expansile cysts; the remaining four had firm masses. All patients remain free of disease without further therapy at 5, 13, 14, 25, 66, and 108 months. Early recognition of this previously unreported and unusual clinical circumstance of a benign teratoma to grow after chemotherapy will allow for surgical salvage. PMID- 6288221 TI - Referral patterns of childhood brain tumors in the state of Connecticut. AB - Data from the Connecticut Tumor Registry (1968-1979) were analyzed to determined whether referral patterns influenced survival rates in children with brain tumors. Two-hundred-seventy-eight children with brain tumors were identified. Less that one-third of the children received all their treatment at university cancer centers. An actuarial analysis of survival rates revealed that children with medulloblastomas treated solely at university cancer centers had projected five-year survival rates of 7.4%. Children with medulloblastomas treated solely at community hospitals had projected five-year survival rates of 29%. Children with brainstem gliomas treated at university cancer centers had projected five year survival rates of 40% whereas of those children treated at community hospitals only one patient was alive at 13 months. No major differences in survival were identified among children with other tumor types. These data suggest that children with certain types of brain tumors may fare better when treated at university cancer centers. PMID- 6288222 TI - Carcinoma of the male breast in West Africans and a review of world literature. AB - Twelve West African men with carcinoma of the breast were treated at the Lagos University Teaching Hospital over a ten-year period, 1971-1980. These cases represent 2.4% of both male and female breast carcinomas treated at the hospital. The mean age was 56.3 years. The left breast was involved 11 times more often than the right. There were no other associated malignancies. Tumor was preceded by gynecomastia in 22% of the cases. Neither trauma, estrogen therapy, nor irradiation appeared to be etiologic factors. The majority of the lesions were advanced. Simple or extended simple mastectomy followed by external radiotherapy was the treatment of choice. There were long delays before patients sought help at a hospital. This was in part because local remedies were tried before resorting to a hospital. The five-year survival rate was 22.2%. The advanced stage of the tumor before surgery rather than race accounted for the low survival rate. PMID- 6288223 TI - Nonseminomatous germ cell cancer of the testis. Reducing treatment-related morbidity in patients with disseminated disease. AB - Fourteen previously untreated patients with metastatic nonseminomatous germ cell cancer of the testis (NSGC) were treated with a modified VAB-4 regimen which was designed to reduce treatment-related morbidity. Nine of 10 patients with minimal disease and the only patient with advanced pulmonary disease achieved a complete response (CR) with chemotherapy alone. Two of three partial responders with advanced abdominal disease were converted to CR status with radiotherapy and/or surgery. None of the 12 CRs have relapsed (median duration of follow-up, 28+ months). We observed no granulocytopenic fever or permanent renal insufficiency. These results indicate that NSGC patients with a low tumor burden can be spared substantial toxicity without adversely affecting complete response rates. PMID- 6288224 TI - Immunohistochemistry of soft tissue tumors. Myoglobin as a tumor marker for rhabdomyosarcoma. AB - Myoglobin, found exclusively in striated muscle, can be considered a fetal antigen. This study investigated its usefulness as a tumor marker for rhabdomyosarcoma (RMS) using an immunoperoxidase technique. Eight-nine percent (89%) or 27 rhabdomyosarcomas contained immunoreactive myoglobin. In contrast, all other soft tissue tumors (54), carcinomas (10), and numerous normal tissues including smooth muscle were negative. Among RMS, staining correlated directly with cytoplasmic differentiation regardless of histologic type. Importantly, six of nine small cell RMS with no initially appreciated cytoplasm were positive, although focally. Since the three negative RMS had only limited tissue available for study, it remains possible that all RMS would exhibit rare positivity if sizeable specimens were tested. Myoglobin should be considered an extremely sensitive and specific tissue tumor marker for rhabdomyosarcoma of considerable clinical importance. PMID- 6288225 TI - Estrogen receptor content and ploidy of human mammary carcinoma. AB - In 46 cases of invasive ductal mammary carcinoma the relationship between estrogen receptor (ER) content and ploidy was investigated. Seventy-one percent of the diploid tumors were ER+ against 40% ER+ tumors in the nondiploid group. Forty percent of the ER+ cases had nondiploid tumors and in several of these more than one tumor cell population could be demonstrated. The results support the hypothesis that multiclonal or mosaic composition of the tumor may explain why 30 45% of the patients with apparently ER+ tumors do not respond to endocrine therapy. PMID- 6288226 TI - Hormone receptors in hepatoblastoma: a demonstration of both estrogen and progesterone receptors. AB - A case of mixed hepatoblastoma occurring in a four-year-old girl is described. Light and electron micrographs illustrate a predominant mesenchymal component and a less conspicuous epithelial element. The tumor contained specific cytosolic protein receptors for estrogen and progesterone. The estrogen receptor titer was 10.5 fmol/mg cytosol protein and the progesterone receptor titer was 62.2 fmol/mg cytosol protein. PMID- 6288227 TI - Patterns of failure in small cell carcinoma of the lung. AB - The initial sites and frequencies of disease progression in 97 patients with small cell carcinoma of the lung treated in a Northern California Oncology protocol were analyzed. Among the extensive disease complete responders (25 patients), the chest was the most frequent initial relapse site (18 patients), followed by the liver (nine patients) and bone (six patients). For those patients who had a partial or no response to treatment, the chest was the most frequent site of persistent disease and the majority progressed in the chest initially. The addition of chest irradiation (5000 rad/5 weeks) to patients with limited disease significantly reduced the incidence of relapse (25%) and prolonged the disease-free interval in the chest in the complete responders, but did not affect the failure pattern in partial and nonresponders. All patients received prophylactic cranial irradiation and three limited disease patients (10%) and three extensive disease patients (4%) progressed in the brain. PMID- 6288228 TI - Extraskeletal osteogenic sarcoma after treatment for Wilms' tumor. AB - A large proportion of children with Wilms' tumor will become long-term disease free survivors. A small number of these children are at risk of developing second malignant neoplasms. There have been no previous reports of osteogenic sarcoma of the chest wall following treatment of Wilms' tumor. Our patient was age seven years when he received surgery, radiation therapy and chemotherapy for Wilms' tumor, eight years when he received radiation and chemotherapy for pulmonary metastases of Wilms' tumor, and 13 years when he developed osteogenic sarcoma of the chest wall. PMID- 6288229 TI - Cytological and cytogenetical studies on human meningioma. PMID- 6288230 TI - Chromosomal breakage and sister chromatid exchange in peripheral blood lymphocytes and lymphoblastoid cell lines in the X-linked lymphoproliferative syndrome. AB - The frequencies of chromosomal aberrations and sister chromatid exchange (SCE) were measured in lymphoblastoid cell lines (LCLs) and in phytohemagglutinin (PHA)- and pokeweed mitogen (PWM)-stimulated lymphocytes from males with X-linked lymphoproliferative (XLP) syndrome, their obligate carrier mothers, and control subjects. We observed an increased frequency of chromosomal aberrations including increased polyploidy in LCLs derived from families with XLP with time in culture. The SCE rate in LCLs (mean of 3.89 SCEs per cell) was much lower than that in PHA or PWM-stimulated lymphocytes: PWM-stimulated lymphocytes showed 9.58 SCEs per cell and PHA-stimulated cells had 11.38 SCEs per cell. A greater number of chromosomal gaps and breaks in the D-group chromosomes of LCLs of affected males and carrier females were identified compared to the number expected, based on chromosomal length and the number of aberrations seen in PHA-stimulated cell cultures. No differences in the frequency of SCEs or chromosomal aberrations were found in control subjects and affected males or carrier females in the peripheral lymphocytes stimulated by PHA. Phenotypes of XLP appear to arise from failure of immune responses to Epstein-Barr virus (EBV) and not from intrinsic chromosomal breakage or instability. PMID- 6288231 TI - Intervening effect of L-ascorbic acid on Epstein-Barr virus activation in human lymphoblastoid cells and its comparison with the effect of retinoic acid. PMID- 6288232 TI - Similarity of Epstein-Barr virus early polypeptides induced by various tumor promoters. AB - Epstein-Barr virus (EBV)-harboring non-producer Raji cells were activated to express Epstein-Barr virus early antigen (EA) by the combined use of n-butyrate and various tumor promoters such as 12-O-tetradecanoyl phorbol-13-acetate (TPA), Euphorbiaceae plant extracts and the toxic microbial metabolite, teleocidin. With regard to the structural differences among the inducing agents (promoters), the patterns of EBV early polypeptides were strikingly similar. A common pathway is probably involved in the activation of the latent genomes of the virus in the host cells. PMID- 6288233 TI - DIfferential ability of Ambystoma tigrinum hepatic microsomes to produce mutagenic metabolites from polycyclic aromatic hydrocarbons and aromatic amines. AB - A number of carcinogenic aromatic amines when activated by liver microsomes from a salamander, Ambystoma tigrinum, are mutagenic for Salmonella tester strains sensitive to frameshift mutagens. However, 2 polycyclic aromatic hydrocarbons (PAH) (benzo[alpha]pyrene (BaP) and perylene) that are rendered mutagenic by mammalian microsomes are not activated by Ambystoma mixed-function oxidases. BaP was chosen for study because it is a well-known environmental carcinogen; perylene, an isomer of BaP, has been implicated as a etiological agent in cutaneous neoplasia in Ambystoma. These results support the observation that amphibians are quite resistant to PAH carcinogenesis and suggest that aromatic amines may be more appropriate model carcinogens. PMID- 6288234 TI - S-Adenosyl-L-methionine is a carbon donor in the conversion of benzo[alpha]pyrene to 6-hydroxymethylbenzo[alpha]pyrene by rat liver S-9. AB - In earlier studies we reported that 6-hydroxymethylbenzo[alpha]pyrene is a metabolite of benzo[alpha]pyrene and of 6-methylbenzo[alpha]pyrene when these substrates are incubated with homogenates or microsomal preparations of liver and microsomal preparations of subcutaneous tissue from Sprague--Dawley rats. However, the origin of the carbon donor was not established. We now report that S adenosyl-L-methionine (SAM) can serve as a carbon donor in the metabolic formation of 6-hydroxymethylbenzo[alpha]pyrene from benzo[alpha]pyrene by the S-9 fraction of rat liver. PMID- 6288235 TI - Corticosteroids and corticotrophin in the treatment of neurologic disorders, with emphasis on neurologic disorders of childhood. PMID- 6288236 TI - Pharmacologic enhancement of neuromuscular transmission in myasthenia gravis. PMID- 6288237 TI - Phase II study of vindesine in patients with advanced breast cancer. AB - Vindesine, a new vinca alkaloid, was administered to 20 patients with advanced refractory breast cancer in a phase II trial. The drug was given at a dose of 3 mg/m2 by iv bolus each week for 6 consecutive weeks, and responding patients were maintained on a dose of 4 mg/m2 every 2 weeks. Nineteen patients were evaluable for disease response; partial remissions were obtained in five patients, for a response rate of 26%. Leukopenia was the major dose-limiting toxic reaction, but most patients were able to tolerate this schedule without difficulty. Neurotoxicity was mild and did not require dose reduction. PMID- 6288238 TI - Dose-response curves for predicting misonidazole-induced peripheral neuropathy. AB - Using pharmacokinetics determined for 27 patients who received iv misonidazole, we have studied the relationship between various pharmacokinetic parameters and the probability of observing peripheral neuropathy. By plotting the probability of observing peripheral neuropathy as a function of each individual parameter and using a linear logistic regression model, we have found a general sigmoid response relating the observations of peripheral neuropathy to the area under the pharmacokinetic curve (AUC), the total cumulative dose throughout the course, the plasma concentration, the plasma elimination half-life, and the total cumulative AUC. By using linear logistic regression analysis, the best correlation with observing peripheral neuropathy was seen with the AUC. Statistical considerations are discussed in depth. This type of analysis may well prove useful for other toxic agents used in cancer treatment. PMID- 6288239 TI - Phase II clinical study of Yoshi 864 in epithelial ovarian carcinoma: A Gynecologic Oncology Group Study. PMID- 6288240 TI - Yoshi 864 plus medroxyprogesterone acetate in adenocarcinoma of the kidney: A Southwest Oncology Group Study. PMID- 6288241 TI - Formation of superoxide and in vitro inactivation of viruses, by hexopyranosid-3 uloses. PMID- 6288242 TI - Caries preventive doses of fluoride and cyclic AMP levels in human plasma. PMID- 6288243 TI - The effect of fluoride on the uptake of albumin by hydroxyapatite. PMID- 6288245 TI - [HLA system and Wilms' tumour--familial study]. PMID- 6288244 TI - [Our first experience of scintiscan in the detection of pericarditis]. PMID- 6288246 TI - [Appearance of radioprotective effect of Gammaphos (WR-2721) in mice]. PMID- 6288247 TI - Distal tubular segments of the rabbit kidney after adaptation to altered Na- and K-intake. II. Changes in Na-K-ATPase activity. AB - Na-K-ATPase activity was measured in the convoluted part of the distal tubule (DCT), the connecting tubule (CNT) and the cortical collecting duct (CCD). The segments were microdissected from freeze-dried kidney tissue of rabbits adapted to various salt diets and exposed to large differences in endogenous and exogeneous mineralocorticoids. The Na-K-ATPase activity in the DCT is not influenced by mineralocorticoids. They do influence the activity in the CNT and in the CCD. In the CNT the highest activity was found with a low Na-, high K diet. At the beginning of the CNT the enzyme activity is higher than in the end portion. While canrenoate-K treatment has no effect on Na-K-ATPase activity in the initial portions of the CNT, this drug decreases the Na-K-ATPase activity significantly in the end portion of the CNT. DOCA treatment has a significant effect on the enzyme activity in the CNT only in the end-portion of the segment, but provokes the highest Na-K-ATPase activity in the CCD. The changes in Na-K ATPase are found to be associated with corresponding changes in the baso-lateral cell-membrane area in the segments affected. PMID- 6288248 TI - Studies of hCG binding and endocytosis in rat ovary by ultrastructural immunocytochemistry. AB - Localization of hCG binding sites and the process of endocytosis in pseudopregnant rat ovaries were investigated by indirect electron-microscopic immunocytochemistry. Immature female rats were treated with pregnant-mare serum gonadotropin (PMSG) and human chorionic gonadotropin (hCG) to induce ovarian luteinization. Eight days after priming with PMSG-hCG and 1-6 h before sacrifice the animals were given another injection of hCG to bind the receptors. Receptor sites to hCG localized by reaction product were present in most luteal cells, but not in primary follicular cells. The receptor sites were distributed on luteal cell surfaces facing interstitial spaces. Endocytotic pits containing hCG binding sites were rarely seen 1 h after hCG injection. At 2h, hCG and presumably its receptor were taken up within endocytotic vesicles with the evidence of reaction product coated on the vesicle wall. With time, fusion of endocytotic vesicles with lysosome occurred and the reaction product appeared in phagolysosomes. The reaction product was localized on phagolysosomal inner surface or in free granular form. These findings suggest that hCG and its receptors were internalized through endocytotic pits and endocytotic vesicles and delivered to lysosomes probably for degradation. An additional experiment for localization of acid phosphatase was also performed to delineate the lysosomes and phagolysosomes. PMID- 6288250 TI - Immunohistochemical demonstration of TSH-, LH- and ACTH-cells in the hypophysis of tadpoles of Xenopus laevis D. AB - By use of the immunofluorescence technique TSH-, LH- and ACTH-cells were localized in the hypophysis of tadpoles of Xenopus laevis. The first signs of the activity of these cells were observed in early stages of the development, i.e., stage 39 for ACTH, and stage 42 for TSH and LH. PMID- 6288249 TI - Effects of acute opiate-peptide administration on pro-opiomelanocortin cells of the intermediate lobe of the rat pituitary. AB - The effects of acute injections of synthetic opiate peptides into the lateral cerebral ventricle of young adult male rats on cells of the intermediate lobe of the pituitary were studied. Met-enkephalin (100/micrograms) injected into anesthetized rats, or 20 micrograms beta-endorphin administered via a previously implanted cannula to unanesthetized animals, will lead to cell degranulation and often to expanded Golgi zones and prominent regions of rough endoplasmic reticulum in secretory cells when tissue is fixed 45--60 min after peptide administration. Treatment of animals with the opiate antagonist naloxone hydrochloride prior to enkephalin injection appeared to prevent the cellular changes elicited with peptide alone. Observations suggest that opiate peptides administered to the cerebrospinal fluid may stimulate release of pro opiomelanocortin-peptide from pituitary cells. PMID- 6288251 TI - Active calcium transport and calcium-dependent membrane phosphorylation in human peripheral blood lymphocytes. AB - The characteristics of the calcium pump were investigated in intact human peripheral blood lymphocytes /PBL/ and in inside-out vesicles prepared from their plasma membranes. Intact PBL were loaded with calcium by a short exposure to A23187 ionophore. After the elimination of the ionophore, calcium-loaded PBL produced an ATP-dependent, external lanthanum sensitive, uphill calcium extrusion. Calcium pump in intact PBL was insensitive to ouabain and /until cellular ATP was provided/ to oligomycin and dinitrophenol. Maximum calcium extrusion rate and the alkali cation sensitivity of the process were similar to those in human red cells. Calcium was partially sequestered by PBL, and this calcium could be released by A23187 ionophore only. Inside-out plasma membrane vesicles prepared from hypotonically lysed PBL showed an ATP + Mg2+-dependent uphill calcium uptake. This calcium transport was insensitive to ouabain, oligomycin, or dinitrophenol, while blocked by lanthanum and quercetin. Calmodulin significantly stimulated calcium pumping in EDTA-washed vesicles. ATP dependent and -independent calcium uptake rates, respectively, showed different calcium concentration dependences. When PBL membrane vesicles were phosphorylated by gamma 32P-ATP, a calcium-induced, hydroxylamine-sensitive incorporation of 32P was found in 120-150 000 molecular weight proteins. Depending on the way of membrane preparation, the molecular weight of the phosphoprotein was shifted. Similarly to that found in red cell membranes, sensitivity to calmodulin stimulation and partial proteolysis of the calcium pump molecule showed an inverse relationship. PMID- 6288252 TI - How cyclic AMP and its receptor protein act in Escherichia coli. PMID- 6288253 TI - Yeast centromere DNA is in a unique and highly ordered structure in chromosomes and small circular minichromosomes. AB - We have examined the chromatin structure of the centromere regions of chromosomes III and XI in yeast by using cloned functional centromere DNAs (CEN3 and CEN11) as labeled probes. When chromatin from isolated nuclei is digested with micrococcal nuclease and the resulting DNA fragments separated electrophoretically and blotted to nitrocellulose filters, the centromeric nucleosomal subunits are resolved into significantly more distinct ladders than are those from the bulk of the chromatin. A discrete protected region of 220-250 bp of CEN sequence flanked by highly nuclease-sensitive sites was revealed by mapping the exact nuclease cleavage sites within the centromeric chromatin. On both sides of this protected region, highly phased and specific nuclease cutting sites exist at nucleosomal intervals (160 bp) for a total length of 12-15 nucleosomal subunits. The central protected region in the chromatin of both centromeres spans the 130 bp segment that exhibits the highest degree of sequence homology (71%) between functional CEN3 and CEN11 DNAs. This unique chromatin structure is maintained on CEN sequences introduced into yeast on autonomously replicating plasmids, but is not propagated through foreign DNA sequences flanking the inserted yeast DNA. PMID- 6288254 TI - On the formation of spontaneous deletions: the importance of short sequence homologies in the generation of large deletions. AB - Using lacl-Z fusion strains of Escherichia coli we have devised systems that detect deletions of varying lengths. We examined deletions 700-1000 base pairs long, and genetically characterized over 250 spontaneous deletions. Of these, we analyzed 24 by direct DNA sequencing and 18 by inspection of restriction fragment patterns. Deletions of this size occur almost exclusively at short repeated sequences in both (recA+ and recA- strain backgrounds, but are detected 25-fold more frequently in a recA+ background. The frequency of deletion formation correlates with the extent of homology between the short repeated sequences, although other factors may be involved. The largest hotspot, which accounts for 60% of the deletions detected, involves the largest homology in the system (14 of 17 base pairs). Altering a single base pair within this homology reduces deletion incidence by an order of magnitude. We discuss possible mechanisms of deletion formation and consider its relationship to the excision of transposable elements. PMID- 6288255 TI - T4 endonuclease VII cleaves holliday structures. AB - T4 endonuclease VII cleaves Holliday structures in vitro by cutting two strands of the same polarity at or near the branch point. The two unbranched duplexes produced by cleavage each contain a strand break that can be sealed by DNA ligase. This suggests that the cut sites are at the same position in the nucleotide sequence in each strand. The joint action of endonuclease VII and DNA ligase can therefore resolve Holliday structures into genetically sensible products. These observations account for the role of endonuclease VII in the DNA metabolism of phage T4, and provide the first example of an enzyme that acts specifically on branch points in duplex DNA. PMID- 6288256 TI - A small subclass of SV40 T antigen binds to the viral origin of replication. AB - We examined the affinities of SV40 large T antigen for unique viral DNA sequences by binding SV40 Bst NI DNA fragments in extracts of infected or transformed cells, and then immunoprecipitating the T antigen-DNA complex. The G fragment, which spans the viral origin of replication (ori) was quantitatively bound to T antigen. A T-antigen-specific monoclonal antibody (McI 7), which recognized only 5%-10% of the T antigen from infected or transformed cells, immunoprecipitated the majority of the ori-binding activity. This suggests that only a minor subclass of wild-type T antigen is active in binding to the origin. C6 cells contain a replication-defective mutant T antigen that when tested in the DNA binding immunoassay, showed no affinity for the ori fragment. McI 7 not only failed to immunoprecipitate ori binding in C6 cells, but also did not detect any labeled C6 T antigen whatever. Thus McI 7 recognizes an immunologically distinct subset of wild-type 7 antigen that comprises the origin-binding form of the viral protein, which is absent in the C6 T antigen population. McI 122, which recognizes a 53 kilodalton host protein that complexes with T antigen, immunoprecipitated ori-binding activity from extracts of infected or transformed cells, but not from C6 cells. Thus wild-type T antigen can bind ori sequences even when complexed to the host protein. These data suggest that T antigen consists of different subpopulations with different functions. PMID- 6288257 TI - Cloning and structural analysis of integrated woodchuck hepatitis virus sequences from hepatocellular carcinomas of woodchucks. AB - Woodchuck hepatitis virus (WHV), like the related hepatitis B virus, induces in its natural host hepatocellular carcinomas that contain integrated viral sequences. As a first step in determining whether and how the integrated sequences contribute to formation of the tumors in which they are found, we have cloned two such integrations of WHV and have determined their structure by restriction mapping and heteroduplex electron microscopy. The identity of the cloned sequences was confirmed by comparison of restriction sites in the clones with those located by Southern blot analysis of tumor DNA. Viral sequences in both integrations are extensively rearranged, and in neither were all parts of the viral genome represented. In this respect, the behavior of WHV in vivo is similar to that of other DNA tumor viruses that have been studied in vitro. We discuss the implications of these results in relation to possible mechanisms for tumor induction by WHV. PMID- 6288258 TI - Detection of a transforming gene product in cells transformed by Moloney murine sarcoma virus. AB - We identified, in cells transformed by Moloney murine sarcoma virus (M-MuSV clone 124), a protein encoded by the M-MuSV transforming gene, v-mos. An antiserum against a synthetic peptide corresponding to the C terminus of a protein predicted from the v-mos nucleotide sequence specifically recognizes a protein doublet of approximately 37,000 daltons from 35S-methionine-labeled M-MuSV 124 transformed producer cells. By peptide mapping, this protein is almost identical to the 37 kd in vitro translation product from the M-MuSV v-mos gene. Immunoprecipitates from 32P-labeled cells contain a single v-mos-specific phosphoprotein, which has at least six sites of phosphorylation containing phosphoserine. Pulse-chase experiments show that the lower band in the 35S methionine-labeled doublet is the primary translation product, which is modified, probably by phosphorylation, to yield the upper band. A similar mos protein is immunoprecipitated from HT1-MuSV-transformed cells, but not from uninfected NIH/3T3 cells. These mos proteins are present at very low levels in transformed cell lines. Cells acutely infected with M-MuSV 124, however, transiently contain much higher levels of the mos protein. These high levels coincide with extensive cell mortality. PMID- 6288259 TI - Nuclear location of the putative transforming protein of avian myelocytomatosis virus. AB - The putative transforming protein of avian myelocytomatosis virus MC29 is a 110,000 dalton (P110gag-myc) polyprotein comprised of sequences derived from both the gag region and the MC29-specific myc region. Two approaches have been taken to determine the location of the MC29 gag-related proteins in transformed cells: subcellular fractionation and immunofluorescence. Analysis of subcellular fractions of MC29-transformed cells by immunoprecipitation indicates that the majority of the gag-myc polyprotein is found in the nuclear fractions of Q8 cells (a nonproducer line of MC29-transformed quail embryo fibroblasts) and nonproducer cells derived from a liver tumor of MC20-infected quail. This is in contrast to the distribution of gag-related helper virus proteins lacking myc, which are found only in nonnuclear fractions of superinfected Q8 cells. The purity of unlabeled nuclei was assessed by electron microscopy and enzyme assays, revealing little contaminating material from other subcellular fractions. Immunofluorescence experiments using monospecific anti-gag serum showed specific, intense immunofluorescence in the nuclei of fixed Q8 cells. In contrast, the majority of P75gag-erb, a candidate transforming protein produced by avian erythroblastosis virus (AEV), is absent from the nuclei of nonproducer AEV transformed chick embryo fibroblasts. The nuclear association of the MC29 transforming protein may be related to some of the unique properties of MC29 transformed cells. PMID- 6288260 TI - Are mitochondrial structural genes selectively amplified during senescence in Podospora anserina? AB - Genetic and transcriptional maps have been constructed for the mitochondrial genome of the Ascomycete Podospora anserina. These data have been plotted on the restriction maps for Sal I, Xho I, Bam HI, Eco RI, BgI II and Hae III. We have characterized and cloned a new and unique senescent mitochondrial DNA (beta-event senDNA) and have organized all of the recognized senDNAs on the genomic maps. We make the observation that all of the known and characterized senDNAs are derived from specific genes or gene regions of the young mitochondrial genome. We have unambiguously assigned the alpha-event senDNA (the 2.6 kb monomer) to the oxi3 gene locus and the beta-event senDNA to the oxi2 gene locus. PMID- 6288261 TI - Chloroplast DNA rearrangements are more frequent when a large inverted repeat sequence is lost. AB - We examined the arrangement of sequences common to seven angiosperm chloroplast genomes. The chloroplast DNAs of spinach, petunia and cucumber are essentially colinear. They share with the corn chloroplast genome a large inversion of approximately 50 kb relative to the genomes of three legumes--mung bean, pea and broad bean. There is one additional rearrangement, a second, smaller inversion within the 50 kb inversion, which is specific to the corn genome. These two changes are the only detectable rearrangements that have occurred during the evolution of the species examined (corn, spinach, petunia, cucumber and mung bean) whose chloroplast genomes contain a large inverted repeat sequence of 22-25 kb. In contrast, we find extensive sequence rearrangements in comparing the pea and broad bean genomes, both of which have deleted one entire segment of the inverted repeat, and also in comparing each of these to the mung bean genome. Thus there is a relatively stable arrangement of sequences in those genomes with the inverted repeat and a much more dynamic arrangement in those that have lost it. We discuss several explanations for this correlation, including the possibility that the inverted repeat may play a direct role in maintaining a conserved arrangement of chloroplast DNA sequences. PMID- 6288263 TI - DNA intermediates in transposition of phage Mu. AB - Transposable genetic elements can insert into DNA sites that have no homology to themselves. Evidence that there is a physical linkage between a transposable element and its target DNA sequence during transposition comes from studies on bacteriophage Mu DNA transposition in which plasmids containing Mu DNA have been shown to attach to host DNA. We report the isolation of key structures, seen after induction of Mu DNA replication, after cloning lac operator into Mu DNA and using the lac repressor-operator interaction to trap Mu DNA on nitrocellulose filters. We have localized Mu sequences within these structures in the electron microscope by visualizing the lac operator-repressor interaction after binding with ferritin-conjugated antibody. This analysis shows that key structures contain replicating Mu DNA linked to non-Mu DNA and that replication can begin at either end of Mu. PMID- 6288262 TI - Directed transposon Tn5 mutagenesis and complementation analysis of Rhizobium meliloti symbiotic nitrogen fixation genes. AB - An 18 kb region adjacent to and surrounding the genes for nitrogenase (nif) was cloned from the genome of the symbiotic nitrogen-fixing species Rhizobium meliloti. A total of 31 Tn5 insertions in the nif region were constructed and assayed for their effect on symbiotic nitrogen fixation (Fix phenotype). Fix- insertions were found in two clusters, one 6.3 kb region not containing essential symbiotic genes. The locations of at least three transcription units containing Fix genes were deduced from complementation analysis between genomic nif::Tn5 insertions and nif::Tn5 insertions on mobilizable cloning vectors. The locations of R. meliloti genes nifH, nifD and nifK, which code for the single subunit of the nitrogenase Fe protein and for the two subunits of the nitrogenase MoFe protein respectively, were determined by DNA hybridization to cloned Klebsiella pneumoniae nif genes and by comparison of partial R. meliloti DNA sequences with K. pneumoniae nif gene sequences. R. meliloti nifH, D and K are located in the 6.3 kb fix-::Tn5 cluster and are transcribed in the order nifH, nifD, nifK, which is the same order as in K. pneumoniae. PMID- 6288264 TI - Genetic properties of chromosomally integrated 2 mu plasmid DNA in yeast. AB - We obtained strains of yeast with large segments of 2 mu plasmid DNA integrated at several chromosomal locations by selecting genetically for recombination between a chromosomal sequence carried on a 2 mu-circle-containing hybrid plasmid and a homologous sequence on the chromosome. In all diploids examined, the presence of 2 mu circle sequences causes a marked instability of the chromosome into which the 2 mu DNA is inserted. Although in some cases the loss of genetic markers is due to physical loss of the entire chromosome, in most cases the loss of markers appears to be due to a mitotic homozygotization of markers: the allelic information from the homologous chromosome replaces the information distal to the integrated 2 mu DNA. The instability caused by integrated 2 mu DNA sequences requires the activity of the specialized site-specific recombination system encoded by the 2 mu plasmid. We propose that the presence of integrated 2 mu DNA allows efficient integration of additional copies of the intact 2 mu plasmid by the action of the plasmid-coded special recombination system. Unequal sister-strand exchanges within the inverted repetition would result in the formation of dicentric chromosomes whose breakage during mitosis might begin a cycle analogous to the breakage-fusion-bridge cycle described many years ago in maize. PMID- 6288265 TI - An altered DNA conformation detected by S1 nuclease occurs at specific regions in active chick globin chromatin. AB - The single-stranded activity of S1-nuclease cleaves globin chromatin in red cell nuclei in specific regions. The cleavages are observed only in tissues in which the globin genes are active, and they "switch" to reflect the switching pattern of globin-gene expression in embryonic and adult red cells. The positions of the S1 cleavages in the beta- and alpha-globin chromatin correspond to the general region of known DNAase I-hypersensitive sites, but can be distinguished in detail. When DNA segments containing these regions are subcloned into pBR322 and the supercoiled molecules are treated with S1, similar sites are cleaved in the purified supercoiled (but not linear) recombinant plasmid DNA. However, the dominant S1 cutting sites are shifted in the plasmid vis-a-vis the chromatin. We believe that some aspect of DNA sequence is translated into an altered DNA structure in chromatin and that it is this altered structure that is recognized by s1 nuclease and possibly by certain chromosomal proteins. Several physical properties reflected in the S1 digestion of supercoiled plasmids suggest a mechanism for generating differences in daughter cells during development. PMID- 6288266 TI - Structure and flanking regions of soybean seed protein genes. AB - We have characterized the structure and flanking region of genes representing two, coordinately expressed, soybean seed protein gene families. One family directs the synthesis of the major storage protean glycinin; the other encodes a 15.5 kd polypeptide of unknown function. DNA blot hybridization experiments showed approximately three, nonallelic genes in the glycinin family and two in the 15 kd protein family, and showed that these families are not selectively amplified or rearranged during embryogeny. R-loop and S1 nuclease mapping studies demonstrated no detectable introns in the 15 kd protein genes but at least one and possibly two in the glycinin genes. No interfamily clustering of these genes occurs within a 10-15 kb chromosomal domain. Nor are they contiguous to other genes expressed at moderate levels during embryogenesis. Each of them, however, is contiguous to a gene expressed at another developmental period in the leaf. These leaf genes encode rare class messages which constitute only 1 X 10(-5%) of the leaf mRNA, or about one molecule per cell. R-loop analysis of two leaf genes showed that one contains no detectable introns while the other possesses at least three. DNA gel blot studies showed that only one of the seed protein genomic clones contains an interspersed repetitive DNA element. Pairwise cross hybridization studies did not detect any flanking sequences shared by the 15 kd protein, glycinin and leaf genes. PMID- 6288267 TI - Functional significance and evolutionary development of the 5'-terminal regions of immunoglobulin variable-region genes. AB - The 5'-terminal sequence of the kappa light-chain gene expressed in MPC11 plasmacytoma cells was definitively determined by analysis of both the kappa mRNA and the gene from which it is transcribed. The distance between the cap site and the translation initiation codon is only 3 nucleotides, and in a minor variant only 2 nucleotides, considerably less than that found in other species of nucleus derived mRNA. S1 nuclease protection experiments with MPC11 nuclear RNA indicate that the cap sites are coincident with the transcriptional start sites, suggesting that the 5'-terminal heterogeneity is caused by imprecision in transcriptional initiation. A comparison of the 5'-terminal structure of the MPC11 V kappa gene with that of several other V genes indicates that the length of the first exon, which is composed of the 5' untranslated region which is composed of the 5' untranslated region and a sequence encoding most of the signal peptide, is highly conserved. Within this set of examples, the 5' untranslated region varies from 3 to 33 nucleotides, and the signal-peptide-coding block from 46 to 76 nucleotides. This analysis has also provided insight into the genetic origins for two anomalous properties of the MPC11 light chain. PMID- 6288268 TI - Duplication and deletion in the human immunoglobulin epsilon genes. AB - The human IgE gene encodes a polypeptide chain that is involved in allergic reactions and in the immune response to parasitic disease in man. We have cloned three chromosomal regions corresponding to this sequence and find that two of them derive from curiously duplicated gene segments that also encode IgA constant region genes. One of the IgE sequences corresponds to the active gene, and its structure defines a complete amino acid sequence of the human IgE constant region. The other cloned segment is a pseudogene from which the first two IgE coding domains have been deleted and replaced by a switch-like sequence that also occurs close to the normal IgE gene. The third IgE segment remains unlinked to the other heavy-chain genes. Evidently, the epsilon-alpha locus has been the site of several complicated genetic rearrangements during recent evolutionary time. PMID- 6288269 TI - Cellular responsiveness to cyclic AMP in a phosphodiesterase-defective mutant of Dictyostelium discoideum. AB - Responsiveness of Dictyostelium discoideum amoebae to cAMP, a chemotactic mediator, was investigated in a strain defective in cAMP-phosphodiesterase production. Cells were subjected to a high cAMP signal (10(-6) M) in the presence or absence of exogenous phosphodiesterase, and the changes of intracellular cAMP and cGMP concentrations and of adenylate cyclase activity were measured. In the presence of cAMP hydrolysis, both adenylate and guanylate cyclases are transiently activated. In the absence of hydrolysis, the high and constant extracellular cAMP concentration is sufficient to elicit a re-activation of adenylate cyclase a few minutes after the first transient response. In contrast, levels of cGMP remain basal for at least 20 min after termination of the initial response to the cAMP addition. PMID- 6288270 TI - Growth arrest of proadipocytes at a distinct predifferentiation G1 state associated with cytotoxic responsiveness to 8-bromo cyclic AMP. AB - The control of cell proliferation can result from the coupling of growth arrest and differentiation. In this regard, we recently demonstrated that growth arrest which precedes the differentiation of 3T3 T proadipocytes must occur at a distinct state in the G1 phase of the cell cycle (GD). Cells arrested at GD differ in several biological parameters from cells arrested in G1 at other states induced by either serum deprivation (GS) or nutrient deficiency (GN). Specifically, GD-arrested cells can differentiate in the absence of DNA synthesis and GD-arrested cells can be induced to proliferate when stimulated with 1-methyl 3-isobutylxanthine; GS- and GN-arrested cells cannot. In addition, GD-, GS- and GN-arrested cells reside at topographically distinct states in G1. We now report that GD-arrested proadipocytes are also distinct in that they are highly sensitive to a cytotoxic effect of 8-bromocyclic AMP, whereas GS- and GN-arrested cells are not. PMID- 6288271 TI - Delayed type hypersensitivity responses to radiation leukemia virus variants. PMID- 6288273 TI - [Juvenile fibroma of the nasopharynx]. PMID- 6288272 TI - [Comparison of efficiency of various methods of induction of Epstein-Barr virus antigens synthesis in human lymphoblastoid cell lines]. PMID- 6288274 TI - [Surgical technics in juvenile nasopharyngeal fibroma supplied by the internal carotid artery]. PMID- 6288275 TI - Studies on 1,2,3,4-tetrahydroisoquinolines. IV. beta-Adrenoceptor activity and absolute stereochemistry of (-)-5,7-dihydroxy-1-(3,4,5-trimethoxybenzyl)-1,2,3,4 tetrahydroisoquinoline. PMID- 6288276 TI - Changes in the plasma membrane of yeast observed by spin label electron spin resonance and caused by photodynamic attack. AB - Spin label electron spin resonance (ESR) was used to characterize the response of lipid regions of the plasma membrane of yeast to photodynamic attack. Following photodynamic attack, the structure of these lipid regions changed resulting in the disappearance of an apparent order-disorder phase transition as well as impeding the diffusion of the steric acid based spin label 12NS into and across the plasma membrane. We propose that singlet molecular oxygen reacting with unsaturated carbon bonds in the fatty acyl chains of lipid surrounding channel proteins leads to an increase in the order of the lipid array and/or a change in the channel protein's conformation and is the cause of the lethal effect of externally sensitized photodynamic action. PMID- 6288277 TI - Public opinion regarding alternative medicine. PMID- 6288280 TI - Benzo[a]pyrene metabolites: formation in rat liver cell-culture lines, binding to macromolecules, and mutagenesis in V79 hamster cells. AB - Benzo(a]pyrene was metabolized in liver cell lines derived from BC-IV and BD-IV rats which included several chemically-transformed lines (IAR-6-1; IAR-19; IAR 28), one spontaneous transformant (IAR-27) as well as one non-malignant line (IAR 20). Cultures were treated with tritiated benzo[a]pyrene over a 5-day period. The cells and medium were extracted with ethyl acetate and the distribution between organic-soluble and water-soluble metabolites determined. Organic-soluble metabolites consisting of dihydrodiols, phenols and quinones were determined by high-pressure liquid chromatography, and macromolecular binding of BP to each cell line was measured over a 24-h period. Comparisons between binding and overall metabolism were not directly proportional in these liver cell lines. However, there was a positive correlation for benzo[a]pyrene mutagenesis in the V 79 hamster cell assay with 8-azaguanine as a marker when the cell lines with the highest (IAR-20) and lowest (IAR-27) metabolic competence were used as activating cell layers. PMID- 6288279 TI - Fibrinolytic activity and morphological transformation of hamster embryo cells. AB - Excretion of plasminogen activator from colonies of Syrian hamster embryo cells has been studied after sequential exposure of the cells to benzo[a]pyrene (3 days), and 12-O-tetradecanoyl-phorbol-13-acetate (3 days). The excretion of plasminogen activator was assayed using the fibrin/agarose overlay technique. The frequency of plasminogen activator-positive colonies was about two times higher for morphologically transformed colonies than for colonies with normal morphology growing on the same dish. Thus, 9% of the transformed colonies, compared to 4% of the colonies with normal morphology, gave clear zones of lysis in the fibrin/agarose overlay after 2 h of incubation. The frequency of plasminogen activator-positive colonies on untreated dishes was 2%. The addition of protease inhibitors strongly reduced the formation of clear zones of lysis, while they did not affect the frequency of morphologically transformed colonies. The data show that the expression of plasminogen activator is not an obligatory event in the process of morphological transformation. PMID- 6288278 TI - [A case of mixed tumor of the lacrimal gland: histologic and prognostic discussion]. PMID- 6288281 TI - Chemical carcinogen activation in the rat mammary gland: intra-organ cell specificity. AB - Cells from 50-55 day old virgin Sprague-Dawley female rat mammary gland were divided into parenchymal (epithelial) and stromal enriched populations. The ability of these cells to activate carcinogens was quantitated employing a mediated mutagenesis assay. In addition, the populations' ability to produce water soluble metabolites from these carcinogens was estimated. We report that the potent mammary carcinogen 7,12-dimethylbenz[a]anthracene was activated by both mammary parenchymal and stromal cells, while the non-mammary carcinogen aflatoxin B1 was not activated by either cell type. However, the weak mammary carcinogen benzo[a]pyrene was activated by the stromal cells and not by the parenchymal cells from which mammary carcinomas arise. The data suggest that the intra-organ relationship between cell types that activate a carcinogen, and cell types that undergo neoplastic transformation, may in part help explain the organ specific potency of a carcinogen. PMID- 6288282 TI - Effect of butylated hydroxyanisole on the metabolism of benzo[a]-pyrene and the binding of metabolites to DNA, in vitro and in vivo, in the forestomach, lung, and liver of mice. PMID- 6288284 TI - Effect of tumor promoters on soft-agar growth of Swiss 3T3 cells infected with SV40 tsA mutants. AB - The availability of many SV40 mutants, in which the ability of the virus to transform fibroblasts is variously affected, has prompted us to investigate the effect of treating SV40-infected cells with known tumor promoters on the expression of the transformed phenotype. Using mouse Swiss 3T3 cells and various SV40 tsA mutants unable to transform these cells at 39 degrees C, we have observed a dramatic effect of the potent phorbol ester promoter, 12-O tetradecanoyl-phorbol-13-acetate, on the formation of macroscopic colonies in soft-agar at the restrictive temperature of 39 degrees C. The efficiency of other phorbol esters and various substances such as anthralin, saccharin, sodium cyclamate, mellitin, griseofulvin and benzoyl peroxide, was in agreement with their reported promoting activities suggesting that mouse Swiss 3T3 cells infected with SV40 tsA mutants could provide a quick and easy test to detect promoters. PMID- 6288283 TI - Inactivation of DNA-binding metabolites of benzo[a]pyrene and benzo[a]pyrene-7,8 dihydrodiol by glutathione and glutathione S-transferases. AB - The binding to DNA of reactive metabolites of trans-7,8-dihydro-7,8 dihydroxybenzo[a]pyrene (BP-7,8-diol) was studied following the incubation of tritiated benzo[a]pyrene (BP) and BP-7,8-diol with nuclei from livers of 3 methylcholanthrene-treated rats. Binding was inhibited to a small extent by glutathione (GSH) alone and to a much greater extent by GSH and cytosol or purified GSH-transferases B and E. In this respect GSH-transferases A and C were also active, but less so. Inhibition of binding of BP-7,8-diol metabolites to DNA mediated by GSH-transferases was associated with the formation of GSH conjugates. The extent of inhibition of binding was similar in incubations of nuclei alone, nuclei and rat liver microsomes, and calf thymus DNA and rat liver microsomes. This indicates that reactive metabolites of BP-7,8-diol, formed either by nuclei or microsomes, are readily accessible to soluble GSH-transferases. GSH and cytosol were also active in inhibiting DNA-binding of reactive metabolites from 9 hydroxybenzo[a]pyrene (9-OH-BP). Thus, in the rat hepatocyte GSH and GSH transferases may be important in protecting DNA from electrophilic attack by reactive BP-7,8-diol and 9-OH-BP species. PMID- 6288285 TI - Intracellular potassium activity in rabbit sinoatrial node. Evaluation during spontaneous activity and arrest. PMID- 6288286 TI - Angiotensin II receptors on human platelets. AB - The investigation of the interaction between angiotensin II and its receptors in human subjects has been hampered by the inaccessibility of human tissue containing angiotensin II receptors. In order to find a more accessible angiotensin II-binding tissue, we studied angiotensin II binding to platelets in normal human volunteers. Platelet preparations purified on Ficoll: Isopaque gradients were incubated with 125I-angiotensin II (30 pm), with and without unlabeled angiotensin at 22 degrees C, separating bound from free hormone by microcentrifugation. Binding was linearly related to the number of platelets incubated, and, at 8 X 10(5) cells/microliters, specific binding ranged from 0.8 to 10%. Scatchard analysis indicated a binding site with a Kd of 2.4 +/- 0.3 X 10(-10) m which agreed well with the Kd by displacement analysis (3.1 X 10(-10) m). The relative binding potencies for angiotensin II and analogues were: angiotensin II = des-Asp1 an angiotensin II greater than [Sar1, Ala8] angiotensin II greater than des-Asp1-[Ile8] angiotensin II greater than angiotensin I. The effect of high and low sodium (Na) intake (200 vs. 10 mEq/day) on platelet angiotensin II binding was studied in nine subjects. Compared to low Na, high Na intake produced an 80% increase in the angiotensin II-binding capacity (P less than 0.01) with no significant change in binding affinity. We conclude that human platelets possess angiotensin receptors whose binding characteristics and modulation by dietary sodium resemble the properties of the receptors on "classical" animal angiotensin target tissues. The platelet may provide an accessible source of angiotensin receptors for a detailed study of angiotensin receptor interaction in human tissue. PMID- 6288287 TI - Beta-endorphin levels in infant apnea syndrome: a preliminary communication. PMID- 6288288 TI - Prevention of adsorption losses during radioimmunoassay of polypeptide hormones: effectiveness of albumins, gelatin, caseins, Tween 20 and plasma. PMID- 6288289 TI - Leucocyte sodium-potassium adenosine triphosphatase and leukemia. AB - Na-K ATPase of healthy leucocytes is relatively low (109 +/- 20 nmol . mg-1 . h 1), as is the permeability of the plasma membrane to cations. In this study of leucocytes from leukaemic patients, the Na-K ATPase was increased. There was an associated increase in membrane permeability to sodium and calcium. In one patient, this led to a clinically dangerous and spurious suggestion of hyperkalaemia. It is suggested that some leukaemic conditions may be associated with 'leaky' membranes. PMID- 6288290 TI - 'Pseudohypertriglyceridemia' caused by hyperglycerolemia due to congenital enzyme deficiency. AB - A 76-year-old man was found to have a false hypertriglyceridemia due to a 40-fold increased glycerolemia. This metabolic change was due to a deficiency in glycerol kinase (ATP:glycerol phosphotransferase, EC 2.7.1.30) activity in the cells of this patient as shown by incubation of his white blood cells with [14 C]glycerol. Several chromatographic analyses and quantitative assays were performed on plasma and urine of this patient and of his relatives. The small number of this family's members did not allow to specify the mode of transmission of this genetic trait. PMID- 6288291 TI - Diagnostic relevance of humoral and cell-mediated immune reactions in patients with acute viral myocarditis. AB - Sera of 177 patients with acute myocarditis (10 coxsackie B 3/4, four influenza, four mumps, 15 cytomegalovirus, 144 undefined) were tested by indirect immunofluorescence for autoantibodies against heart and skeletal muscle and vital or air-dried adult cardiocytes. Antibody-dependent cytolysis, lymphocytotoxicity and antibody-dependent cellular lymphocytotoxicity were assessed using viral adult rat cardiocytes as target cells. Muscle-specific anti-sarcolemmal antibodies of the anti-myolemmal type--often associated with non-organ-specific anti-endothelial antibodies--were demonstrated in nine out of 10 patients with coxsackie B, in all patients with influenza and mumps and in 65 out of 144 patients with undefined myocarditis. In contrast, 13 out of 15 patients with cytomegalovirus myocarditis lacked anti-sarcolemmal antibodies but had low titre anti-inter fibrillary antibodies instead. In the presence of complement, anti myolemmal antibodies induced cytolysis of vital cardiocytes, whereas hepatocytes remained unaffected. Titres of anti-myolemmal antibodies correlated with the degree of cardiocytolysis. The anti-myolemmal immunofluorescent pattern and the cytolytic serum activity could be absorbed with the respective viral antigens suggesting that these antibodies cross-react with moieties of the virus itself and may be both diagnostic and aetiological markers in acute viral myocarditis. Lymphocyte-mediated cytotoxicity against heterologous cardiac target cells could not be observed in our patients with myocarditis of proven viral aetiology. However, lymphocyte-mediated cytotoxicity was demonstrated in 10 ASA-positive and one ASA-negative patient with myocarditis of unknown origin. ASA-positive sera blocked lymphocytotoxicity in three of these patients. PMID- 6288292 TI - Epstein-Barr virus (EBV)--lymphoid cell interactions. II. The influence of the EBV replication cycle on natural killing and antibody-dependent cellular cytotoxicity against EBV-infected cells. AB - We investigated the influence of the Epstein-Barr virus (EBV) replication cycle on natural killing (NK) activity and antibody-dependent cellular cytotoxicity (ADCC) against EBV-infected cells. Peripheral blood lymphocytes from healthy EBV seropositive and -seronegative donors were separated on Ficoll-Hypaque gradients and used as effector cells in the standard 51Cr release assay to measure NK and ADCC. EBV-genome positive RAJI and DAUDI cells superinfected with either the non transforming P3HR-1 EBV or the transforming B95-8 EBV were used as targets. The results obtained show that most normal individuals have ADCC and NK activity against P3HR-1 EBV-infected RAJI cells. Both the cytotoxic activities increased with the proportional increase in effector/target (E/T) ratios, assay incubation time, dose of the infecting virus and the time of pre-infection with EBV. Moreover, the data obtained indicate that different immune mechanisms are effective at different stages of the virus replication cycle. During the early stages of virus replication, EBV-superinfected cells are more susceptible to ADCC than to NK, whereas in later stages the susceptibility to NK is increased significantly and appears to play a more dominant role. The nature of the target cells or the strain of EBV used to superinfect these targets did not influence their susceptibility to ADCC and NK activity; however some quantitative differences were found. Using metabolic inhibitors such as cytosine arabinoside, phosphonoacetic acid, actinomycin D, cycloheximide and puromycin, it was found that new DNA synthesis is not essential but some RNA and protein synthesis is necessary, late in the viral cycle, for the superinfected cells to become susceptible to NK and ADCC. PMID- 6288293 TI - Calcium antagonism: a new approach. AB - 1. The calcium antagonists (calcium entry blockers or slow channel inhibitors) are a heterogenous group of substances that inhibit the voltage-activated inward displacement of Ca2+. 2. These compounds can be subdivided according to their chemistry (organic and inorganic) and their preferred sites of action (myocardium, vascular or nodal tissue). 3. Some of the substances which inhibit slow channel transport exhibit other properties that may be of pharmacological significance. PMID- 6288294 TI - Effect of different Epstein-Barr virus-determined antigens (EBNA, EA, and VCA) on the leukocyte migration of healthy donors and patients with infectious mononucleosis and certain immunodeficiencies. PMID- 6288295 TI - Pepstatin A-induced degranulation and superoxide anion generation by human neutrophils. PMID- 6288296 TI - The effects of corticosteroids on the antigen presenting properties of human monocytes and endothelial cells. PMID- 6288297 TI - T-lymphocyte cycling in human cyclic neutropenia: effects of lithium in vitro and in vivo. PMID- 6288298 TI - The isolation and functional activity of polymorphonuclear leukocytes and lymphocytes separated from whole blood on a single percoll density gradient. PMID- 6288300 TI - Survival trends in women with epithelial cancer of the ovary in Northern Alberta 1960-1979. PMID- 6288299 TI - Polypeptide hormone receptors: their role in the pathogenesis of human disease. PMID- 6288302 TI - In vitro studies of bone formation and resorption. PMID- 6288301 TI - Case report: management of methotrexate toxicity in an anephric patient. AB - We describe the case of an anephric woman on chronic hemodialysis who developed breast cancer with positive axillary nodes. After mastectomy, we attempted to treat her with adjuvant chemotherapy. She suffered severe toxicity involving the skin, gastrointestinal tract and bone marrow after a single dose of 15 mg (10 mg m-2) of methotrexate. She received high doses of leukovorin and hemodialysis was continued. Although hemodialysis caused little acute change in the serum methotrexate levels, over a period of 3 weeks the levels did decrease, her symptoms of toxicity subsided, and she recovered. The case demonstrates the limited effectiveness of hemodialysis for treating methotrexate toxicity and suggests that leukovorin is useful for reversing it. PMID- 6288303 TI - Anti-parkinson drugs in the Batten-Spielmeyer-Vogt syndrome; a pilot trial. PMID- 6288304 TI - The neurology of hypertension, a neurologist view. PMID- 6288307 TI - Significance of basal ganglia calcifications. PMID- 6288305 TI - X-linked congenital hydrocephalus. AB - In this report we describe a Dutch family with ten cases of X-linked recessive congenital hydrocephalus with a high perinatal mortality. In three cases necropsy has confirmed the diagnosis. In the best documented case the most striking features are absence of obstruction or stenosis of the aqueduct and congenital malformation of the cerebral cortex. On the basis of our findings and on reviewing the literature, the hypothesis is put forward that the defective gene on the X-chromosome is responsible for a pathological influence on cerebral cortex development and extraventricular CSF pathways. The expressivity of the genetic defect may be variable, causing extreme phenotypic variants (CHC and/or MR) under the influence of the different modifying genetic or environmental factors. Genetic counselling is difficult in families with no X-linked CHC precedent, since the mutant gene rather produces a communicating HC, secondarily complicated by narrowing of the aqueduct, and as at present there is no way of detecting beforehand heterozygote carriers. PMID- 6288306 TI - A case of chorea due to polycythaemia vera. PMID- 6288308 TI - Adverse effects of metrizamide. PMID- 6288309 TI - First experiences with proton spin tomography (NMR). PMID- 6288310 TI - Ataxic hemiparesis: clinical, electrophysiologic, radiologic and pathologic observations. PMID- 6288311 TI - Tc-99m diphosphonate uptake in primary malignant cystosarcoma phyllodes. PMID- 6288313 TI - Antenatal detection of recurrence of Majewski dwarf (short rib-polydactyly syndrome type II Majewski). AB - Two cases of Majewski dwarfism are reported. both were the product of the same non-consanguinous marriage and each exhibited the key features of this syndrome: polysyndactyly, short ribs, cleft lip, disproportionately short tibiae and a normal pelvis. The first infant, a baby boy, died shortly after birth, whilst the second was detected prenatally by ultrasound and had female characteristics. Several skeletal dysplasias have been demonstrated in utero by ultrasound, but we believe this to be the first reported case of a Majewski dwarf detected by this method. In view of the difficulty in diagnosis of this syndrome comparison is made with other short rib-polydactyly syndromes. PMID- 6288312 TI - Clinical comparison of cimetidine and ranitidine. AB - Ranitidine is a histamine H2-receptor antagonist that differs structurally from cimetidine. We assessed its kinetics, potency, and duration of effect in patients with chronic duodenal ulcers. Ranitidine had an absorption lag time of approximately 30 min, an absorption half-life (t 1/2) of approximately 40 min, and an elimination t 1/2 of 3 hr, all differing from those of cimetidine. It is five times as potent, so that equal doses of ranitidine induce considerably longer suppression of both basal and food-stimulated gastric acid secretion than cimetidine. PMID- 6288314 TI - Liver metastases in Wilms' tumour. AB - Fifty patients with Wilms' tumour have been reviewed to evaluate the incidence of liver metastasis in order to identify the optimum time and frequency for radiological investigation. Liver involvement occurred in 12% of our cases. The most rewarding periods for the diagnosis of liver metastasis are the first few months of the illness, the period immediately after stopping chemotherapy, or any time that lung metastases are identified. Liver scintigraphy is recommended as the initial investigation, and abnormal cases should proceed to ultrasound or computed tomography. PMID- 6288315 TI - Ultrasonic patterns in trophoblastic disease. PMID- 6288316 TI - The natriuretic hormone: recent developments. PMID- 6288317 TI - Vitamin D from skin: contribution to vitamin D status compared with oral vitamin D in normal and anticonvulsant-treated subjects. AB - 1. The plasma 25-hydroxycholecalciferol [25-(OH)D3] response to measured u.v. irradiation applied thrice weekly for 10 weeks was investigated in normal and in anticonvulsant-treated subjects. 2. Levels of plasma 25-(OH)D3 achieved after u.v. irradiation were similar in both normal and anticonvulsant-treated subjects, suggesting that hepatic microsomal enzyme induction does not lead to low plasma 25-(OH)D3 concentrations. 3. Cholecalciferol was present in plasma of normal subjects in a very low concentrations (less than 5.0 nmol/l) and did not increase until plasma 25-(OH)D3 levels exceeded 62.5 nmol/l. 4. Cholecalciferol occurred in significant concentrations in plasma during whole body u.v. irradiation or during oral dosage of 62.5 nmol (100 i.u) or more daily. 5. Plasma 25-(OH)D3 concentrations reached a steady state after 5-6 weeks of u.v. irradiation or of oral intake within the usual intake range. 6. Cholecalciferol synthesis in skin calculated from the steady-state equation was 0.0015 +/- 0.0008 nmol/mJ. 7. Cholecalciferol synthesis in skin was also calculated from the oral dosage required to yield the same plasma 25-(OH)D3 concentration as u.v. irradiation and was 0.0024 +/- 0.0018 nmol/mJ. 8. Rates of cholecalciferol synthesis calculated from these data suggest that many of the population of England receive insufficient u.v. irradiation to maintain vitamin D status throughout the year. PMID- 6288318 TI - Inherited complement deficiency states and SLE. PMID- 6288319 TI - Evaluation of coronary artery disease by exercise stress testing including radionuclide studies. PMID- 6288320 TI - Computed tomography in the diagnosis of pituitary adenoma. PMID- 6288322 TI - Raised concentration of a circulating Na+-K+-ATPase inhibitor in essential hypertension. PMID- 6288321 TI - Exchangeable and total body sodium and potassium in various hypertensive syndromes. PMID- 6288323 TI - The mechanisms of hypercalciuria are unnecessary for treatment of recurrent renal calcium stone formers. PMID- 6288324 TI - Primary hepatocellular carcinoma and hepatitis B virus. PMID- 6288325 TI - Ketoconazole in the treatment of chronic mucocutaneous candidiasis secondary to autoimmune polyendocrinopathy-candidiasis syndrome. PMID- 6288326 TI - Chromosomal instability of SV40-transformed human prostatic epithelial cell lines. AB - Three SV40-transformed derivatives (G1, T2, and T5) of human prostatic epithelial cells were analyzed karyotypically. For comparison, an SV40 derivative (TA) obtained from isogenic fibroblasts, was also studied. The chromosome complements of these cells, as well as a sub-clone of T2 isolated in soft agar (T2-A5), were analyzed using banding techniques. Numerical as well as structural changes were observed in all transformed cultures. Karyotypic changes in all cells at a given passage level appeared to be random. On the other hand, characteristic differences in modal chromosome number, and type and number of abnormal chromosomes were observed among the different lines. Most cells of two of the three epithelial lines (T1 and T2) were either hypo- or pseudodiploid, whereas T5 consisted of a mixed hypodiploid and hypotetraploid population. The TA subline was also predominantly hypo- and pseudodiploid. Dicentrics, telomeric associations, translocations, and loss of chromosomes were the most prominent abnormalities. The loss of chromosome 18 was characteristic for all epithelial lines. All T1 and T5 cells had lost either one or both copies of the 18. While individual cells of the original T2 line had random karyotypes, most of T2-A5 cells had a relatively uniform karyotypic pattern. They also had a similar pattern of abnormal chromosomes. These observations suggest that culture in soft agar may have selected a particular chromosomal variant. We conclude that transformation of prostatic epithelial cells by SV40 may bring about site specific as well as random chromosomal changes. These changes could reflect either intermediate or sequential stages in progression to neoplasia. PMID- 6288327 TI - Status of prezygotic chromosome lesions in relation to cancer. AB - There are currently three recognized prezygotic chromosome defects that are associated with specific cancers. These are a deletion of band 13q14 with retinoblastoma, deletion of band 11p13 with Wilms' tumor associated with aniridia, and a reciprocal translocation of chromosomes 3 and 8 with clear cell cancer of the kidney. The two deletions have been assumed to be variants of the hereditary subgroup of tumors. There are many obvious similarities between retinoblastoma associated with deletion of 13q13 and Wilms'-aniridia associated with deletion of 11p13, but there are also significant disparities. Each time a deletion of 13q14 has been found it is associated with a retinoblastoma, while deletions of 11p13 have been associated with nephroblastoma in only 4 of 11 cases. When bilateral disease occurs it is usually seen simultaneously in both eyes with retinoblastoma, but with bilateral nephroblastoma the appearance of the first and second tumors is separated by a year or more. These disparities indicated the possibility that more than one etiologic mechanism may operate even within this group of specific chromosomal mutations associated with specific tumors. PMID- 6288328 TI - Calcium channel antagonists in coronary artery spasm and bronchial spasm. What do variant angina and bronchial asthma have in common? PMID- 6288329 TI - Agar (disk) diffusion test susceptibility of clinical isolates of Pseudomonas aeruginosa to azlocillin, cefotaxime, cefsulodin, lamoxactam, mezlocillin, and piperacillin. AB - The standardized Bauer-Kirby agar diffusion test served to examine 100 clinical isolates of Pseudomonas aeruginosa against various recently introduced broad spectrum penicillins and cephalosporins. Neither cefotaxime nor lamoxactam displayed significant activity against this microorganism. Azlocillin, cefsulodin, and piperacillin were significantly more effective (p less than 0.0001) than mezlocillin against the majority of isolates. When compared individually, azlocillin and piperacillin displayed comparable in vitro activity; the same was true for cefsulodin compared with piperacillin. On the other hand, cefsulodin was somewhat more active than azlocillin (p less than 0.05, greater than 0.01) against P. aeruginosa. These data should enable diagnostic laboratories to curtail the number of antimicrobial drugs routinely utilized to examine clinical isolates of P. aeruginosa for antibiotic susceptibility, i.e., piperacillin exclusively. PMID- 6288332 TI - Asbestos and silica: their multiple effects on the lung. PMID- 6288333 TI - [Activity of systems of superoxide radical formation and detoxication in hepatic carcinogenesis]. PMID- 6288330 TI - Chromosomal arrangement of heat shock locus 2-48B in Drosophila hydei. AB - cDNA, copied from nuclear RNA isolated from heat shocked Drosophila hydei cells, has been cloned. From this collection of clones a clone, N09-15, with a 450 bp insert has been isolated that hybridizes in situ to the heat shock locus-2-48B of Drosophila hydei. The N09-15 sequence is present in two different genomic arrangements, as shown by restriction mapping, in our wild type D. hydei population. These genomic arrangements are allelic. Both alleles contain multiple copies of the N09-15 sequence but differ in their lengths and in the distribution of Msp I and Taq I sites. PMID- 6288331 TI - Adrenocortical responsiveness to the synthetic ACTH 1-17 analogue given at different circadian stages. AB - The study concerns the adrenocortical glucocorticoid responsiveness to the ACTH 1 17 analogue given at different times of the day, namely near to the zenith and the nadir of the circadian curve of plasma cortisol. Two schedules of administration of the heptadecapeptide have been performed: a pulse i.v. injection of 4 microgram; b. i.m. injection of 100 microgram. Both the doses were given to the same subject in the morning and in the evening of different days. The chrono-sensitivity of the adrenal cortex to ACTH 1.17 analogue is well evident after pulse stimulation by a micro-dose of the heptadecapeptide; in fact the plasma cortisol increase from basal values is significantly higher in evening than in the morning (p less than 0.001). The cortico-stimulatory effect of the higher dose of ACTH 1.17 analogue lasts about 12 h. and then plasma and urinary glucocorticoids take the usual circadian pattern again. PMID- 6288334 TI - [Determination of the localization of EcoRII restrictase and methylase genes on recombinant plasmids]. PMID- 6288335 TI - [EcoRV restriction of glucosylated DNA of T-even phages and the cloning of the fragments obtained in pBR322]. PMID- 6288336 TI - Effects of acute cannabis use and short-term deprivation on plasma prolactin and dopamine-beta-hydroxylase in long-term users. AB - Six long-term cannabis users (10 - 44 years of use) were studied for four days during which cannabis smoking was followed by three days deprivation and smoking on the fourth day. Blood samples were taken at - 30, + 30 and + 120 minutes from each smoking and once in every deprivation day. Pulse rate and blood pressure were measured before each blood sample. Prolactin and dopamine-beta-hydroxylase activity were estimated in plasma. On the third deprivation day, both prolactin and dopamine-beta-hydroxylase were significantly elevated compared to the pre smoking values, but returned to them after the second cannabis use. The mean arterial pressure followed the same course. The data suggest that cannabis use in man reduces noradrenergic and enhances dopaminergic activity, while deprivation has the opposite effect, possibly through presynaptic receptor mechanisms. PMID- 6288337 TI - [Cefotaxim for peri-operative short-term prophylaxis of infection in cardiac surgery. Concentrations in myocardial, pericardial, fat and skeletal muscle tissues]. AB - Cefotaxim was administered intravenously in a dose of 50 mg/kg body-weight before and immediately after cardiac surgery. The drug's concentrations were measured in serum, myocardium, peri-operative fat, pericardium and skeletal muscle. The highest concentration was found 15 minutes after injection: 32 micrograms/g in pericardium, 22 micrograms/g in myocardium, 17.9 micrograms/g in fat and 12.3 micrograms/g in skeletal muscle. After 2 hours the average concentration in pericardium was 10.7, in myocardium 3.8, in fat 5.4 and in skeletal muscle 2.4 micrograms/g. The corresponding mean serum levels were 237.4 and 33.3 micrograms/ml, respectively. The tissue concentrations were thus high enough to be effective against infections caused by cefotaxim-sensitive organisms. There was primary wound healing in all 16 patients. No other infections, such as pneumonia or urinary infections, occurred. PMID- 6288338 TI - [Cefotaxim--an alternative in the treatment of purulent meningitis in children?]. AB - Cefotaxim was administered to ten children with purulent meningitis. All isolated micro-organisms were highly sensitive to it in serial dilution, except for one case, and were quickly removed from blood and CSF by the administration of cefotaxim alone. Blood and CSF concentrations measured in eight children were much above the minimal inhibitory concentration for the particular micro organism. Thus cefotaxim has a high antibacterial activity and satisfactory CSF passage in purulent meningitis caused by sensitive, especially gram-negative, bacteria and is well suitable for therapy, if penicillins alone or combined are not applicable or effective. PMID- 6288340 TI - [Systemic antimycotic treatment]. PMID- 6288339 TI - [Fatal complication of intravenous administration of synthetic adrenocorticotrophin]. AB - Synthetic 1-24adrenocorticotrophin (tetracosactid, Synacthen) is used in clinical medicine for diagnostic and therapeutic purposes. Side effects may occur due to unphysiological stimulation of endogenous adrenocortical hormone production. Allergic reactions to tetracosactid are rare. Anaphylactic shock leading to death was observed after intravenous injection of tetracosactid in a 14-year-old girl with bronchial asthma. Adrenocorticotrophin antibodies could be demonstrated after death in serum and ascitic fluid using the double antibody method. PMID- 6288341 TI - Gastric carcinoma in a tropical African population. PMID- 6288342 TI - Estrogen synthetase stimulation by dibutyryl cyclic adenosine 3',5'-monophosphate in human choriocarcinoma cell culture: the relative biosynthetic rate of the nicotinamide adenine dinucleotide phosphate (reduced form)-cytochrome c reductase component. PMID- 6288343 TI - Angiotensin II receptors and prolactin release in pituitary lactotrophs. AB - Logical properties of angiotensin II receptors in the rat adenohypophysis were analyzed in cultured rat pituitary cells incubated with angiotensin II and known stimuli of pituitary hormone secretion. PRL release during incubation for 3 h with 3 nM angiotensin II was consistently increased by 68 +/- 5%, comparable with that elicited by TRH (63.1 +/- 4%). The ED50 of 0.5 nM for PRL release by angiotensin II was significantly lower than that of TRH (2.9 nM) in the same cell cultures. The antagonist analog [Sar1,Ala8]angiotensin II prevented the angiotensin-induced rise in PRL production but not that evoked by TRH, whereas dopamine and SRIF inhibited basal, angiotensin, and TRH-stimulated PRL release. Angiotensin II also caused a small increase in ACTH release but had no effect on the release of LH, TSH, and GH. Angiotensin II binding and PRL release were measured in partially purified lactotrophs prepared by elutriation, by which the initial cell suspension was separated into seven fractions. Most of the lactotrophs were present in the two fractions eluted at flow rates of 15.7 and 19.8 ml/min, as indicated by their immunoreactive PRL content. The 2.5- to 3.2 fold enrichment of lactotrophs was accompanied by a 2- to 3.5-fold increase in angiotensin II receptor concentration, with no change in binding affinity (Ka = 3.5 x 10(9) M-1). In the same fractions, angiotensin II-induced PRL release was similarly increased by 1.6- to 3.5-fold above basal, compared with values of less than 1 in the initial cell suspension and other fractions. The preferential location of angiotensin II receptors in the lactotroph-containing fractions and the close correlation between angiotensin II binding sites and stimulation of PRL release indicate the functional importance of the pituitary angiotensin II receptor sites. These findings also suggest that angiotensin II could contribute to the physiological regulation of PRL secretion. PMID- 6288344 TI - Binding and internalization of a luteinizing hormone-releasing hormone agonist by rat gonadotrophic cells. A radioautographic study. PMID- 6288345 TI - Regulation of 21-hydroxylase activity by steroids in cultured bovine adrenocortical cells: possible significance for adrenocortical androgen synthesis. PMID- 6288346 TI - Tooth pulp stimulation potentiates the adrenocorticotropin response to hemorrhage in cats. AB - Stimulation of the maxillary tooth pulp nerve (TP), a predominantly nociceptive afferent fiber system, was assessed for its effect on peripheral plasma ACTH in chloralose-urethane anesthetized cats. These results were compared to those after a transient 10 ml/kg hemorrhage (H), a submaximal neurogenic stressor for ACTH release, and to H plus TP in combination. TP alone for 3 min had no significant effect on ACTH. However, TP during H greatly potentiated the increase in plasma ACTH concentration compared to that seen after H alone. The TP potentiation of the H-induced rise in ACTH was not accompanied by altered cardiovascular responsiveness nor by differences in plasma norepinephrine or glucose relative to that seen after H alone. The data indicate that nociceptive and baroreceptor afferents share a common neural substrate for selective facilitation of ACTH release, but do not interact to potentiate several other physiological responses, such as sympathetic efferent activity. Furthermore, under the conditions of these experiments, selective nociceptor activation in the anesthetized cat is not an adequate stimulus for the release of ACTH. PMID- 6288347 TI - Time- and dose-dependent effect of triiodothyronine on submaxillary gland epidermal growth factor concentration in adult female mice. PMID- 6288348 TI - Corticotropic peptides in the human fetal pituitary. PMID- 6288349 TI - Nuclear 3,5,3'-triiodothyronine receptor concentration increases during deoxyribonucleic acid synthesis in partially synchronized GC cell cultures. PMID- 6288350 TI - Androgen biosynthesis in developing ovarian follicles: evidence that luteinizing hormone regulates thecal 17 alpha-hydroxylase and C17-20-lyase activities. PMID- 6288351 TI - Prolactin inhibits pituitary luteinizing hormone-releasing hormone receptors in the rat. PMID- 6288352 TI - Effect of 3,5,3'-triiodothyronine treatment on potassium efflux from isolated rat diaphragm: role of increased permeability in the thermogenic response. PMID- 6288353 TI - Effect of gonadectomy and gonadal steroid replacement on pituitary and plasma beta-endorphin levels in the rat. PMID- 6288354 TI - Biological effects of an ectopic growth hormone-releasing peptide in cultured adenohypophyseal cells: comparison with growth hormone-releasing activity of porcine hypothalamus. PMID- 6288355 TI - Desensitization to parathyroid hormone in the isolated perfused canine kidney: reversal of altered receptor-adenylate cyclase system by guanosine triphosphate in vitro. PMID- 6288356 TI - Adrenal innervation may be an extrapituitary mechanism able to regulate adrenocortical rhythmicity in rats. AB - The daily rhythm of plasma corticosteroid concentration was studied in nonhypophysectomized rats and in hypophysectomized rats implanted with beeswax pellets containing ACTH and T4. In the first experiment, male hypophysectomized rats were placed on a 12-h light, 12-h dark cycle (onset of light, 0600 h) with food and water available ad libitum. Beeswax pellets containing 1.5 mg ACTH and 150 micrograms T4 were implanted sc in these rats. Beginning 4 days after the implantation of ACTH and T4, daily rhythms of plasma corticosteroid concentration were detected in these rats on 3 successive days. The injection of sodium pentobarbital (40 mg/kg BW) or atropine sulfate (10 mg/kg BW) ip at 1200 h blocked the expected rise in plasma corticosteroid concentration at 1800 h in both these rats and a group of intact rats. In the second experiment, adrenal innervation was disrupted by spinal cord transection at the T-7 level. L-1 transected rats served as operated controls, and a third group was maintained as unoperated controls. One week after surgery, daily rhythms of plasma corticosteroid concentration were present in both unoperated and L-1 controls (P less than 0.01) but not in the T-7-transected rats. Inasmuch as an extrapituitary mechanism was capable of maintaining adrenocortical rhythmicity in hypophysectomized rats and disruption of adrenal innervation suppressed adrenocortical rhythmicity, it was hypothesized that adrenal innervation may be an extrapituitary mechanism which has a role in adrenocortical rhythmicity. PMID- 6288357 TI - Regulation of DNA polymerase beta in rat adrenal gland by adrenocorticotropic hormone. AB - Analysis of extracts of rat adrenal glands showed that the activity of DNA polymerase beta was preferentially decreased upon hypophysectomy, whereas that of DNA polymerase alpha or gamma stayed relatively constant. The adrenal polymerase beta from hypophysectomized rats sedimented at 5-7S through sucrose gradients, whereas that from sham-operated controls sedimented at 3.3-3.6S. The large form of DNA polymerase beta was stable in 0.1 M KCl or 0.5% Nonidet P-40 but was converted into a 3.3S form in 0.5 M KCl. These changes in adrenal DNA polymerase beta were reversed by the daily injection of adrenocorticotropic hormone but were not reversed by exogenous glucocorticoid. PMID- 6288358 TI - Prolactin receptors in luteinized rat ovaries: unmasking of specific binding sites with detergent treatment. PMID- 6288359 TI - Corticotropin releasing factor (CRF): effects on the release of pro opiomelanocortin (POMC)-related peptides by human anterior pituitary cells in vitro. AB - The effect of synthetic ovine corticotropin-releasing factor (oCRF), alpha melanotropin (alpha-MSH) and arginine-vasopressin (AVP) on the release of pro opiomelanocortin (POMC)-related peptides (beta-endorphin, beta-lipotropin, adrenocorticotropin, and the 12K N-terminal) was studied in primary cultures of human anterior pituitary (HAP) cells. The peptides released into the medium were measured by specific radioimmunoassays. HAP cells responded to oCRF at concentrations as low as 10 ng/ml by significantly increasing peptide secretion (p less than 0.005). AVP was also effective, but, it is only 0.1% as potent as oCRF. alpha-MSH was ineffective even at a dose of 10 micrograms/ml. Immunoreactive (IR)-beta EP, IR-beta LPH, IR-ACTH, and IR-HNT were released in approximately equimolar ratios. Partial characterization of the peptides released by HAP cells with gel chromatography on Sephadex G-50 superfine revealed two peaks of IR-beta EP (as beta EP and beta-LPH), two peaks of IR-beta LPH (as beta LPH and gamma-LPH), one peak of IR-ACTH, and two peaks of IR-HNT. In conclusion, oCRF is a potent secretagogue for the equimolar release of ACTH, HNT and related peptides in the human pituitary gland. PMID- 6288360 TI - Protein kinase activity of purified Leydig cells: low protein kinase activity causes impaired steroidogenesis by band two cells. AB - Using a 0-32% continuous metrizamide density gradient, interstitial cells could be separated into five distinct bands. Cells localized in bands 1 (B1), 2 (B2), and 3 (B3) were isolated and incubated for 1h with or without human chorionic gonadotropin (hCG). Both B2 and B3 cells responded to hCG with increased cyclic AMP formation, but only B3 cells produced significantly more testosterone. Protein kinase activity of B2 cells was found to be extremely low compared with B1 and B3 cells. Additional treatment of B3 cells with collagenase did not cause any change in protein kinase activity. These results indicate that decreased protein kinase activity may be responsible for impaired testosterone synthesis in B2 cells. PMID- 6288362 TI - Endogenous substrates for cAMP-dependent phosphorylation in dispersed bovine parathyroid cells. AB - Agonists that increase intracellular cAMP in bovine parathyroid cells ((l) isoproterenol (ISO), dopamine, prostaglandin E2) as well as cAMP analogs (dibutyryl cAMP and 8-bromo-cAMP) stimulated the phosphorylation of 2 endogenous proteins with apparent molecular weights of 19K and 15K. The time course and concentration-dependence of ISO-stimulated phosphorylation in these cells correlated well with known effects of ISO on intracellular cAMP and PTH release. ISO-stimulated phosphorylation of these two proteins was rapidly reversed by (l) propranolol. Although 2 mM extracellular calcium inhibited ISO-stimulated PTH secretion, it did not significantly affect the phosphorylation of the 19K and 15K bands. These results are consistent with a physiologic role for the phosphorylation of these proteins in agonist-stimulated PTH secretion in bovine parathyroid cells. PMID- 6288361 TI - Glucocorticoid regulation of 1,25(OH)2vitamin D3 receptors: divergent effects on mouse and rat intestine. AB - We have investigated the possibility that glucocorticoids alter responsiveness to vitamin D by regulating the 1,25(OH)2D3 receptor in rat intestine. In contrast to the mouse where glucocorticoids caused receptors to decline, rats treated with glucocorticoids showed a substantial increase ( approximately 50%) in the number of intestinal 1,25(OH)2D3 receptors. This resulted from an increase in receptor content with no change in affinity for 1,25(OH)2D3. Receptor stimulation was even greater in vitamin D-deplete rats. Moreover, adrenalectomy led to a significant decline in receptor number. Although the properties of the receptor are similar in rat and mouse intestine, the divergent response to glucocorticoids emphasizes major differences between species in the regulation of 1,25(OH)2D3 receptor number. PMID- 6288363 TI - Rat Leydig cell phospholipid content is increased by luteinizing hormone and 8 bromo-cyclic AMP. PMID- 6288364 TI - Presence of corticotropin releasing factor-like immunoreactivity in hypophysial portal blood. AB - Hypophysial protal blood was collected from pentobarbital anesthetized male rats, and the content of corticotropin releasing factor-like immunoreactivity (CRF-LI) was measured using an RIA developed for synthetic ovine CRF. The concentration of CRF-LI in portal plasma was 104.9 +/- 9.7 pM. This concentration is in the range which has been shown to stimulate ACTH and beta-endorphin secretion in vitro. These data support the hypothesis that a molecule with the same or similar structure as synthetic ovine CRF is a physiologically significant hypothalamic releasing factor. PMID- 6288365 TI - Effect of exogenous ACTH on the circadian rhythm of cortisol and number of leukocytes in the peripheral blood of sheep. PMID- 6288366 TI - Cushing's syndrome: problems in management. AB - CS comprises a group of disorders characterized by hypercortisolism. The variety of causes--pituitary-dependent CS (CD), adrenal tumor, and the ectopic ACTH syndrome--necessitates a variety of therapies--surgical, radiotherapeutic, and medical. Once a specific diagnosis is made, specific therapy can be instituted. Although some controversy persists regarding treatment, particularly that of CD, for most patients it is straightforward. However, in our experience with more than 60 patients, therapeutic dilemmas can arise in a number of circumstances, e.g. the patient with the radiologically normal sella or recurrent CD after adrenalectomy. In addition, the treatment of such conditions as the large ACTH producing pituitary tumor, Nelson's syndrome, the malignant ectopic ACTH syndrome, and adrenal carcinoma is not entirely satisfactory. Our approach to these problems is illustrated by seven cases, and we emphasize that the proper management of CS requires both correct diagnosis and the logical application of all available therapies. PMID- 6288369 TI - Thymidine incorporation by lung fibroblasts as a sensitive assay for biological activity of asbestos. PMID- 6288367 TI - The role of lipoproteins in steroidogenesis and cholesterol metabolism in steroidogenic glands. PMID- 6288368 TI - Controlling cadmium in the human food chain: a review and rationale based on health effects. PMID- 6288370 TI - Effect of cyclic nucleotide analogs on intrachain site I of protein kinase isozymes. AB - The effects of numerous cAMP analogs present in the [3H]cAMP binding reaction on subsequent dissociation of [3H]cAMP from the regulatory subunit of cAMP-dependent protein kinase I and II were analyzed. Certain analogs with modification at either C-8 or C-2 showed relative selectivity for one (site 1) of two intrachain cAMP binding sites of both isozymes. Modification at C-6 caused selectivity for the second site (site 2). The combination of a site-1-directed and site-2 directed analog inhibited [3H]cAMP binding much more than did either analog alone. In general, there was a correlation between the site 1 selectivity and the ability of the analog to stimulate the binding of [3H]cIMP, which selects site 2. The site-1-directed analogs stimulated the initial rate of [3H]cIMP binding. The stimulatory effect was enhanced in the presence of a polycationic protein such as histone and was inhibited by high ionic strength. The type I and II isozymes exhibited large differences in analog specificity for this effect. For type I, of the analogs tested the most efficacious for stimulating [3H]cIMP binding were those containing a nitrogen atom attached to C-8, 8-aminobutylamino-cAMP being the most effective. Type II responded best to analogs containing a sulfur atom attached to C-8, 8-SH-cAMP being the most effective of those tested. The stimulatory effect was accentuated in the presence of MgATP when using type I, but this nucleotide had no effect when using type II. It is proposed that in intact tissues cAMP binding to site 1 of either isozyme stimulates the binding to site 2. PMID- 6288371 TI - Thyroidal cAMP-dependent protein kinases. Particular of the type I kinase, and compartmentalization of the two isoenzymes. PMID- 6288372 TI - The extents of formation of cobalt(II)-radical intermediates in the reactions with different substrates catalysed by the adenosylcobalamin-dependent enzyme ethanolamine ammonia-lyase. AB - 1. The reactions of the adenosylcobalamin-dependent enzyme, ethanolamine ammonia lyase, with the 'good' and 'relatively poor' substrates 2-aminoethanol and (S)-2 aminopropanol respectively, under conditions of saturation with substrate were investigated by rapid freezing in conjunction with electron paramagnetic resonance (e.p.r.) spectroscopy and by stopped-flow spectrophotometry. 2. In disagreement with earlier reports [Babior et al. (1972) J. Biol. Chem. 247, 4389 4392], it was found that the reaction of 2-aminoethanol gave an e.p.r. signal observed in rapid freezing experiments characteristic of a coupled Co(II)-free radical system. This signal was similar to, though not identical with, that obtained with (S)-2-aminopropanol. The steady-state level of the signal with 2 aminoethanol as substrate was 0.56 of that attained with (S)-2-aminopropanol. 3. The results of these e.p.r. experiments were shown to be consistent with stopped flow data obtained under closely similar reaction conditions, the latter indicating a corresponding ratio of 0.64. The results also are consistent with those of a rapid wavelength scanning, stopped-flow spectrophotometric study [Hollaway et al. (1978) Eur. J. Biochem. 82, 143-154]. PMID- 6288373 TI - An intracellular modulator of the 3'5'-cGMP hydrolysis in growing Dictyostelium discoideum amoebae. AB - A heat-stable and acid-stable macromolecular factor present in the cytosol of growing Dictyostelium discoideum amoebae affects specifically the intracellular cGMP phosphodiesterase. It decreases the V of the enzyme but does not alter its Km. It has no effect on the cAMP or cGMP hydrolysis catalyzed by the intracellular cAMP-cGMP phosphodiesterases or by the extracellular phosphodiesterase. It is also expressed in a mutant (HPX235), defective in the synthesis of the cAMP-cGMP phosphodiesterases but capable of intracellular transduction of the chemotactic signal. This factor is resistant to several nucleases, proteases and phospholipases, and has an apparent molecular weight between 3500-10000. In contrast, the protein phosphodiesterase inhibitor secreted by the amoebae exerts an opposite inhibition on the intracellular phosphodiesterases. These two inhibitory factors may regulate intracellular cGMP hydrolysis during the chemotactic response. PMID- 6288375 TI - Yeast mutant defective in synthesis of phosphatidylinositol. Isolation and characterization of a CDPdiacylglycerol--inositol 3-phosphatidyltransferase Km mutant. AB - 1. A Km mutant of Saccharomyces cerevisiae with a lesion in CDPdiacylglycerol inositol 3-phosphatidyltransferase was isolated. The mutant required a high concentration of myo-inositol for growth. 2. The CDPdiacylglycerol-inositol 3 phosphatidyltransferase in the mutant cells showed an apparent Km for myo inositol over 200-times higher than that of the enzyme in wild-type cells. The maximum velocity of the mutant enzyme was comparable to that of the wild-type enzyme. 3. In mutant cells, labelled myo-inositol, phosphate and acetate were incorporated into phosphatidylinositol at much slower rates than in wild-type cells. The phosphatidylinositol content of mutant cells was markedly lower than that observed in wild-type cells. 4. Genetic analysis showed that the growth phenotype of the mutant arose from a single nuclear gene mutation in a gene coding for CDPdiacylglycerol-inositol 3-phosphatidyltransferase. 5. The mutant showed a normal level of phosphatidylserine synthase activity. The phosphatidylserine synthase gene was located between ura3 and hom3 on chromosome V, whereas the CDPdiacylglycerol-inositol 3-phosphatidyltransferase gene showed no linkage with ura3. 6. Labelled acetate was incorporated into various lipids including triacylglycerols, diacylglycerols, sterol esters and phospholipids other than phosphatidylinositol at faster rates in mutant cells than in wild-type cells. Incorporation into both the fatty acid and the sterol moieties was facilitated in the mutant. 7. A striking change in the cell-division process was observed when phosphatidylinositol synthesis was limited. The results showed that phosphatidylinositol synthesis is involved in the cell-division cycle of yeast. PMID- 6288374 TI - Purification of bovine kidney and heart pyruvate dehydrogenase phosphatase on Sepharose derivatized with the pyruvate dehydrogenase complex. AB - Pyruvate dehydrogenase phosphatase has been purified to apparent homogeneity from mitochondrial extracts of both beef heart and beef kidney. An essential step in this three-step purification is affinity chromatography of a largely purified phosphatase fraction using Sepharose beads to which pyruvate dehydrogenase complex is covalently bound through the lipoic acid residues of the dihydrolipoyl transacetylase component of the complex. The purified phosphatase, which has a native relative molecular mass, Mr, of about 140000, is composed of two nonidentical subunits of Mr 89000 and 49000. PMID- 6288376 TI - Phosphorylations of the subcellular matrix in cells transformed by Rous' sarcoma virus. AB - The transforming protein of Rous' sarcoma virus (RSV) is a phosphoprotein of Mr 60 000 (pp60src) which displays protein kinase activity specific for tyrosine residues; pp60src is associated with the plasma membrane and is recovered in the detergent-insoluble material which represents the subcellular matrix of the cell. After phosphorylation of this material of RSV-transformed cells with [gamma 32P]ATP, five phosphoproteins have been detected which are not seen in normal cells. These proteins (Mr = 135 000, 125 000, 75 000, 70 000, 60 000) contain phosphotyrosine. Their phosphorylation is strongly inhibited by anti-pp60src antibodies. In cells transformed by a temperature-sensitive mutant of RSV, these phosphoproteins, present at the permissive temperature, are no longer detected at the non-permissive temperature. It is concluded that these phosphorylations are mediated by pp60src protein kinase activity. This supports a possible role of the phosphorylation of cytoskeletal proteins in the transformation process. PMID- 6288377 TI - Tuftsin analogs for probing its specific receptor site on phagocytic cells. AB - Six new analogs of the phagocytosis-stimulating peptide tuftsin were synthesized with the eventual aim of characterizing and isolating the tuftsin receptor. These analogs can be classified as follows: (a) photoaffinity labelling analogs for the specific covalent attachment to the tuftsin receptor: (b) fluorescent analogs containing either rhodamine or dansyl fluorescent probes for microscopic visualization of the tuftsin receptor; (c) biotin analog for separation and purification of the receptor by affinity methods. In this paper we describe the various synthetic pathways employed to introduce sensitive prosthetic groups into the tuftsin molecule while preserving its biological activity. Activities of the various analogs synthesized as compared to tuftsin in biological and receptor binding assays are described. All analogs are able to stimulate phagocytosis of the macrophage cell as well as complete specifically for tuftsin binding sites on these cells. PMID- 6288378 TI - Transfer of Torpedo acetylcholine receptors to mouse L-cell surface membranes by liposomes containing Sendai virus envelope proteins. PMID- 6288379 TI - A correlative study on the topographical distribution of the receptors for low density lipoprotein (LDL) conjugated to colloid gold in cultured human skin fibroblasts employing thin section, freeze-fracture, deep-etching, and surface replication techniques. AB - The topographical distribution of the receptors for low-density lipoprotein (LDL) conjugated to colloidal gold on cultured human skin fibroblasts was studied using a surface replication technique following critical point drying. In this correlative study, also employing thin sectioning, freeze-fracture and deep etching techniques in conjunction with the polyene antibiotic filipin, it could be shown that LDL preferentially binds to certain microdomains of the plasma membrane, the so-called coated pits. The coated pits can be recognized after filipin treatment both on the P face and E face of the membrane, due to the fact that they are free of filipin-sterol complexes. Filipin was also of value in clearly delimiting the structure of nascent coated pits from their surroundings in surface replicas. Using filipin has the effect of increasing the contrast between the smooth surface of a coated pit and the rough surface of the remaining membrane caused by formation of filipin-sterol complexes. This study has shown that there are differences in the topographical distribution and number of gold particles from fibroblast to fibroblast and even between different areas of the same cell. The surface replication technique using goldlabelled LDL provides s suitable method for improving the interpretation of the spatial arrangement of the LDL receptors. The advantages of this technique are discussed in comparison with the methods previously employed to visualize LDL receptors. PMID- 6288380 TI - Effects of Ca2+ deficiency, collagenase, and mechanical dispersion on the ultrastructure of cardiac myocytes of adult rats. AB - A new perfusion medium for isolating cardiac myocytes from adult rats was developed, thereby yielding numerous viable cells with few morphological changes. The main factors in the isolation procedure are Ca2+ deficiency, collagenase, and mechanical dispersion. Their effects on the ultrastructure of cardiac myocytes were separately tested. In isolated hearts, perfusion with a medium containing a physiological Ca2+ concentration (2.5 mM, controls) preserved the cellular fine structure well, whereas perfusion with a medium containing 2.5 mM Ca2+ plus 0.05% collagenase caused swelling and disruption of most cells. Perfusion with a Ca2+ deficient medium followed by a medium with a low Ca2+ concentration (25 microM) either containing or lacking collagenase resulted in widening of the T-tubules, reduced electron density of the external lamina and occasional separation, or even dissolution of this layer. Some cells were damaged and hypercontracted. These appeared more numerous in suspensions, that means after mechanical dispersion of the myocardium. However, most of the isolated cells were regularly shaped (up to 30-60 min as shown in another study) and their ultrastructure was only slightly altered. This corresponds to an adequate preservation of the cell membranes proven in earlier membrane transfer studies. PMID- 6288381 TI - Stannous ion quantitation in 99mTc-radiopharmaceutical kits. AB - A simple and inexpensive method for the estimation of stannous ion, Sn (II), in radiopharmaceutical kits is described. The method used is a potentiometric titration of Sn (II) in 1 N HCI medium, using potassium iodate as the oxidizing agent in an atmosphere of nitrogen. The apparatus includes pH meter, a platinum electrode, and a simple titration cell. Several commonly used radiopharmaceutical kits were analyzed for their Sn (II) content using this method. These studies indicate that the procedure can be used, as a routine quantitative test for the Sn (II) content of various 99mTc-labeled radiopharmaceuticals. PMID- 6288382 TI - Biochemical studies of the renal radiopharmaceutical compound dimercaptosuccinate. II. Subcellular localization of 99Tc-DMS complex in the rat kidney in vivo. AB - In order to extend and confirm our previous studies of in vivo behaviour of 99mTc DMS in rat kidney cells, the intracellular localization of 99Tc-DMS complex in rat kidney tissue was examined. Animals were injected IV with 99Tc-DMS solution (37 kbq in 1.0 ml) 2 h before killing. Dissected kidneys were homogenized and submitted to differential centrifugation to obtain organelles. The following radioactivity distribution in relation to the total radioactivity of homogenate was obtained in nine experiments: nuclei 3.30% (+/-1.27), mitochondria 9.48% (+/ 2.17), microsomes 7.03% (+/-2.34) and cytosol 57.45% (+/-8.01). The subcellular distribution pattern of 99Tc-DMS was in very good agreement with the pattern of 99Tc-DMS binding to kidney cell organelles obtained by the same procedure. Soluble cytoplasmic proteins binding 99Tc-DMS were excluded from a Sephadex G-25 column with the void volume. Their electrophoretic mobility after agarose gel electrophoresis corresponded to beta-lc- and alpha-2-macroglobulins. The results obtained show unambiguously that the investigated complex penetrates into the kidney cells where it binds mostly to soluble cytoplasmic proteins and mitochondria, and to a lesser extent to both microsomes and nuclei. In another series of six experiments, isolated kidney cortical tissue was used, the same procedure was repeated and similar results were obtained. PMID- 6288383 TI - Biochemical studies of the renal radiopharmaceutical compound dimercaptosuccinate. III. Subcellular distribution and interaction of 99Tc-DMS complex with macromolecules in rat kidney homogenates in vitro. AB - Rat kidney tissue homogenates, incubated in vitro with 99Tc-DMS were subjected to differential centrifugation. Specific radioactivities of subcellular fractions were as follows: cytosol 13,552, microsomes 3,600, mitochondria, 2,707 cpm/mg protein, and nuclei, 2,642 cpm/mg DNA. The cytosol fraction was further analyzed by ion-exchange chromatography on DEAE-Sephadex A-25 column. In this way it was separated into several distinct protein zones, three of them bearing a significant amount of 99Tc-DMS preparation. The highest specific radioactivities of proteins were eluted from the DEAE-Sephadex column with 30 mmol/l and 50-110 mmol/l NaCl solution. Total nucleic acids isolated from purified nuclei, mitochondria and microsomes contained a certain amount of bound 99Tc-DMS complex. Nuclear nucleic acids had the highest specific radioactivity, suggesting that 99Tc-DMS complex penetrates into the nuclei. PMID- 6288384 TI - Massive bilateral nephroblastomatosis in a 13-year-old-girl. PMID- 6288385 TI - A case report of congenital hypomyelination. AB - A 3 year, 11 month-old Japanese male with congenital hypomyelination is described. Clinical features are delay of motor development, generalized muscle hypotonia and weakness, absent tendon reflexes due to peripheral neuropathy, and normal mental development. Electrophysiologically, nerve conduction velocities could not be measured. Histological examination of the right sural nerve revealed total of absence myelin of most of the myelinated fibers. Electronmicroscopically, there was a concentric network of lamellae formed by double-layered sheets of basement membranes with fragments of Schwann cell cytoplasm around the myelinated fibers, so called "onion-bulbs". These peculiar features were similar to those in the cases reported by Lyon, (1969); Kennedy et al., (1977); Karch et al., (1975); and Anderson et al. (1973). PMID- 6288386 TI - Blink reflexes elicited by electrical, acoustic and visual stimuli. I. Normal values and possible anatomical pathways. AB - Blink reflexes elicited by electrical stimulation of the supraorbital nerves, by monoaurally presented 1 kHz sine wave bursts of 20-ms duration, and by flashes of light presented monocularly were investigated in 32 healthy controls. Absolute latencies of the individual components and side differences were measured. An outline of possible anatomical pathways which might be involved in the transmission of these reflexes is given. PMID- 6288387 TI - Age-dependent changes in central somatosensory conduction time. AB - Cervical and cortical somatosensory evoked potentials(SEPs) were recorded in 45 normal subjects. Absolute peak latencies and latency differences between the components P7, N9, N11, N13, P17 and N20 were measured. Subjects aged 40-60 years had significantly longer latencies of N13 and N20 than subjects aged 15-39 years. Moreover, statistical analysis revealed a significant prolongation of N9-N13, N11 N13 and N13-N20 transit times in older subjects. Possible connections with known morphological age-related findings are discussed. PMID- 6288388 TI - Effect of cardiac glycosides on the release of adenylate kinase into cerebrospinal fluid of patients with cerebral arteriosclerosis. AB - 48 patients with cerebral arteriosclerosis were found to have a manifest release of adenylate kinase (AK) into cerebrospinal fluid (CSF). This release was most probably due to an increased leak in the brain cells subsequent to a lowered adenylate charge potential followed by a diminished electrochemical potential in these cells suffering from disturbed oxygen supply. A further increase of AK release into CSF was noted for the 22 patients receiving cardiac glycosides compared to the 26 patients not treated with these drugs. The mean AK value of the former group was 0.119 +/- 0.028 U/l compared to that of the latter group, being 0.089 +/- 0.025 U/l, and this difference was significant (p less than 0.001). The effect of cardiac glycosides is most probably explained by an additional lowering of the membrane electrochemical potential in brain cells of these patients due to the direct action of cardiac glycosides on the Na+- and K+ dependent ATPase system in these cells, resulting in an increased leak in the plasma membrane. PMID- 6288389 TI - The bone scan as a tumour marker in prostatic carcinoma. AB - The technetium methylene diphosphate bone scan was elevated in 100 consecutive new patients presenting with carcinoma of the prostate. 48% of the patients has a positive bone scan at the initial diagnosis. The scan was more helpful than the skeletal X-ray in the diagnosis of bone metastases: 23% of the X-ray-negative patients were scan-positive. Serial bone scans were more sensitive than either X rays or serum acid phosphatase in following the progress of the disease. It is concluded that the bone scan in patients with prostatic carcinoma can be used as a reliable tumour marker, especially for monitoring the course of metastases. PMID- 6288390 TI - VP16-213 in combined modality treatment of small cell carcinoma of the lung. AB - Thirty-four previously untreated patients with histologically proven small cell carcinoma of the lung were treated with a combined modality therapy program that incorporated VP16-213, an epipodophyllotoxin derivative, into the chemotherapy regimen. Initial therapy for two cycles was with V-CAM, VP16-213, cyclophosphamide, doxorubicin and methotrexate. Following two cycles of V-CAM each patient received radiation therapy consisting of 4000 rads to the primary site, both hila and the mediastinum, as well as 2000 rads as prophylaxis to the whole brain. After a one-week rest period the patients received monthly cycles of V-CAM until death. Of 10 patients with stage IIIM0 disease, 7 had a complete response (CR), 1 a partial response (PR) and 2 had progressive disease. The median survival was still not reached by approximately 18 months. Of 24 patients with supraclavicular and/or metastatic disease there were only 5 patients with a CR, 11 with a PR and 8 with progressive disease. Their median survival was approximately 9 months. The 70% overall response rate and 9.3-month median survival of the entire group are essentially the same results as those in previously reported studies. There appears to be no additional benefit when VP16 213 is incorporated into our combined modality program. PMID- 6288391 TI - Augmented immunogenicity of Lewis lung carcinoma by infection with herpes simplex virus type 2. AB - In vitro, LLT cells sustain HSV-2 replication without evidence of lysis. Simultaneously, multiplication of the cells is stimulated. These xenogenized cells were tested for their immunopotentiating capacity: three-step immunization with xenogenized viable cells conferred significantly augmented transplantation resistance to a challenge graft with 4 x 10(4) intact LLT cells. Latency periods preceding tumor formation were increased and 15% of the mice failed to form primary tumors. Metastasis was likewise decreased and 25% of the mice had healthy lungs. Immunopotentiation, however, did not suffice to significantly protect against a challenge with 6 x 10(4) intact cells. The presence of virus-specific neoantigens on HSV-2-infected viable cells was demonstrated by the progressively increasing number of rejections of 4 x 10(4) xenogenized cells during the successive immunization steps. Immunization with non-viable LLT cells did not augment resistance to challenge. PMID- 6288392 TI - Phenotypic differences between tumor cells derived from different stages of neoplastic growth. AB - Under conditions employed in our laboratory, tumors which are induced by avian sarcoma virus (ASV) usually grow progressively for several weeks and then regress. In order to further understand the basis for tumor regression in this model, we compared avian sarcoma cells which were cultured from tumors at different stages of development in terms of various phenotypic properties. The results indicate that tumor cells which are derived from progressively-growing sarcomas are rapidly growing, produce large quantities of the enzyme plasminogen activator, and have much in common generally with chicken embryo fibroblast (CEF) cells that have been transformed by ASV. In contrast, tumor cells that are obtained from regressors have elevated levels of hexose transport, grow very slowly, are greatly enlarged and display properties that are characteristic of senescent cells in culture. PMID- 6288393 TI - Enhanced transformation of mouse embryo cells by SV40 virus following treatment with fibroblast growth factor. PMID- 6288394 TI - Impact of air quality in exercise performance. PMID- 6288395 TI - Dependence of the cardiovascular effects of pindolol on the individual sympathetic nervous activity. PMID- 6288396 TI - Effects of enprofylline, a xanthine lacking adenosine receptor antagonism, in patients with chronic obstructive lung disease. AB - Enprofylline, a xanthine-derivative shown experimentally to lack universal adenosine receptor antagonism, has been examined in patients with partly reversible, chronic, obstructive lung disease. Significant bronchodilation was produced by enprofylline 2 mg/kg, giving a peak plasma concentration of 3.0 +/- 0.6 microgram/ml (mean +/- SD). A dose of 2 + 4 mg/kg dilated the bronchi at least to the same extent as theophylline 9.2 +/- 0.9 mg/kg (plasma level 18.5 +/- 4.7 micrograms/ml). Neither at the low nor at the high dosage (2 +/- 4 mg/kg), giving plasma concentrations of 8.5 +/- 1.4 microgram/ml, did enprofylline produce theophylline-like CNS effects, such as restlessness and tremor, but it did exhibit some of the innocuous side effects expected with xanthine derivatives, such as epigastric discomfort and headache. The comparison with theophylline was limited because different dosage forms had to be used (solution an tablets), which for example, resulted in different absorption rates. Nevertheless, the present findings indicate enprofylline to be potent bronchodilator in patients with obstructive lung disease, suggesting that adenosine-receptor antagonism is not involved in the bronchodilator effects of xanthines. PMID- 6288398 TI - Adenosine receptors in capillaries and pia-arachnoid of rat cerebral cortex. PMID- 6288397 TI - Immature B cells in fetal development and immunodeficiency: studies of IgM, IgG, IgA and IgD production in vitro using Epstein-Barr virus activation. AB - Epstein-Barr virus has been used as a B cell mitogen to explore the parallels between the B cells found in patients with hypogammaglobulinemia and the immature B cells in fetal tissues. Peripheral blood lymphocytes from 29 cases of late onset hypogammaglobulinemia (common variable immunodeficiency) and from 10 cases of X-linked hypogammaglobulinemia were depleted of T lymphocytes and stimulated with virus in vitro. Immunoglobulin production was measured over a 4-week culture period using inhibition radioimmunoassays for IgM, IgG, IgA and IgD. The results were compared with those seen with fetal liver cells, cord blood lymphocytes and adult lymphocytes. Virus-stimulated cells from fetal sources produced small amounts of IgG and IgA relative to IgM, the ratio of IgM to IgG in the second week being in all cases greater than 10. Similar patterns were seen in 25/29 cases of late onset hypogammaglobulinemia, and in the eight cases of X-linked hypogammaglobulinemia that responded in vitro. In contrast, the ratio of IgM to IgG was always less than 8 in cultures of normal adult peripheral blood or bone marrow lymphocytes, and also in cultures from four cases of hypogammaglobulinemia known independently to have abnormal circulating suppressor cells. Eight cases of selective IgA deficiency showed reduced IgA production; six of these showed a normal ratio of IgM to IgG production. Thus, the B lymphocytes which circulate in many patients with hypogammaglobulinemia are functionally immature. PMID- 6288399 TI - Functional subsensitivity of alpha 2-adrenoceptors accompanies reductions in yohimbine binding after clorgyline treatment. PMID- 6288400 TI - Opiate antagonists reverse the centrally mediated antihypertensive action of propranolol in spontaneously hypertensive rats. PMID- 6288401 TI - Pharmacological characterization of presynaptic alpha-adrenoceptors modulating [3H]noradrenaline and [3H]5-hydroxytryptamine release from slices of the hippocampus of the rat. AB - The experiments served to characterize the receptors mediating the inhibitory effect of alpha-adrenergic drugs on K+ (20 mM)-induced [3H]noradrenaline (NA) and [3H]5-hydroxytryptamine ([3H]5-HT) release from slices of the dorsal part of rat hippocampus. Dose-response curves were constructed using the cumulative dose response technique (Frankhuyzen and Mulder, 1981). All of the adrenergic agonist drugs examined inhibited the K+-induced [3H]NA release. NA appeared to have the highest intrinsic activity followed by adrenaline. Clonidine and adrenaline had similar intrinsic activities, while that of oxymetazoline was lowest. The highest pD2 values were observed for oxymetazoline and clonidine, being slightly higher than that of adrenaline followed by NA. By far the lowest pD2 values was observed for phenylephrine. With the exception of phenylephrine, all of the agonists also inhibited the K+-induced [3H]5-HT release. NA, adrenaline and oxymetazoline appeared to have similar intrinsic activities, while that of clonidine was considerably lower. The pD2 values of NA and adrenaline were not significantly different but were somewhat lower than those of oxymetazoline and clonidine. Similar antagonistic effects of phentolamine and yohimbine were observed with respect to the adrenergic inhibition of K+-induced [3H]NA and [3H]5-HT release. Prazosin, however, appeared to be ineffective in both instances. It is concluded from these results that the presynaptic adrenergic inhibiton of [3H]NA as well as [3H]5-HT release is mediated by alpha 2-adrenoceptors located on noradrenergic and serotonergic varicosities, respectively. Furthermore, our data suggest that these alpha 2-adrenoceptors are not pharmacologically identical. PMID- 6288402 TI - Characterization of the opiate receptor in the guinea-pig ileal mucosa. AB - Several opioid peptides and narcotic drugs reduced transepithelial potential difference (PD) and short circuit current (Isc) in guinea-pig ileal mucosa measured in vitro in Ussing chambers. [D-Ala2,D-Leu5]enkephalin was the most potent peptide tested. Enkephalin analogues with altered C-terminal amino acids were less potent, as were beta-endorphin and dermorphin. Etorphine produced potent effects whereas morphine and SKF 10,047 were inactive. Ethylketazocine produced a biphasic dose-response curve. When added by themselves diprenorphine and naloxone produced small increases in Isc. This effect was not seen when Cl- and HCO3- in the Ringer were replaced by SO42-. Diprenorphine and naloxone were able to shift the dose response curves for all agonists to the right, with the exception of that for ethylketazocine. Diprenorphine was a more potent antagonist than naloxone. SKF 10,047 also acted as a pure antagonist. Morphine and ethylketazocine had no antagonist effects. It is concluded that the opiate receptor in the guinea-pig ileal mucosa is similar to a delta-opiate receptor as defined by ligand binding studies, but that some differences also exist. PMID- 6288403 TI - Novel phenylpiperidine opioid antagonists and partial agonists: effects on fluid consumption. AB - The effects of five opioid antagonists, a racemate partial agonist and its agonist and antagonist optical isomers were studied on deprivation-induced drinking. All compounds had a phenylpiperidine nucleus. The antagonists produced dose-related decreases in drinking, and the potencies for decreasing drinking correlated with morphine-antagonist doses. The racemic partial agonist and its agonist isomer decreased drinking at doses higher than those which produced marked analgesia. Within the class of phenylpiperidine drugs studied, some had less specificity than naloxone for the mu-receptors as compared to the delta receptor, but the suppression of drinking was not related to changes in mu-to delta ratios. PMID- 6288404 TI - Mechanism of extraneuronal adrenaline release induced by K+-free medium and ouabain, but not by high KCl in vas deferens of guinea-pig and rat. AB - The effects of K+-free medium, ouabain or high KCl on the contractile response and spontaneous efflux of adrenaline were analyzed in denervated guinea-pig or rat vasa deferentia preloaded with 3 x 10(-5) M (+/-)-adrenaline. The K+-free medium-induced contraction of guinea-pig tissues was related to the concentration of adrenaline used during preloading and was suppressed by the addition of 2.5 mM Rb+ but not by 2.5 mM Cs+. The contractile effects of K+-free medium and 10(-5) M ouabain, but not the response to high KCl, were prevented by 5 x 10(-6) M phentolamine or 3 x 10(-5) M deoxycorticosterone. These contractions, therefore, appear to be mediated by adrenaline released from the extraneuronal compartment. The K+-free medium- or ouabain-induced contractions of denervated rat vasa deferentia were small in comparison with those of the guinea-pig tissue. The amount of spontaneous efflux of adrenaline from the guinea-pig preparation was significantly enhanced by omission of K+ or by 10(-4) M ouabain but not by 80 mM KCl. K+-free medium and 3 x 10(-5) M ouabain merely elicited a small membrane depolarization accompanied by a slight decrease of membrane resistance in the guinea-pig vas deferens, whereas 30 mM KCl markedly altered these electrical parameters. From these findings, it is suggested that the rapid increase in extraneuronal amine release following inhibition of Na+-K+-ATPase may be due to some direct effect of K+ deficiency or of ouabain on the binding between the enzyme and the catecholamine rather than to alterations of membrane properties as a consequence of enzyme inhibition. PMID- 6288405 TI - Differential effect of humoral endorphin on the binding of opiate agonists and antagonists. AB - Humoral (H) endorphin, a novel endogenous factor with opiate-like activity was further characterized using various radioreceptor assays. H-Endorphin displaced both labelled opiates (morphine, ethylketazocine) and opioid peptides (leucine enkephalin) from their specific binding sites in rat brain membranes. The effect of sodium ions on H-endorphin indicates its agonistic nature which is in agreement with previous pharmacological experiments. However, H-endorphin potentiated the binding of the opiate antagonist naloxone to brain membranes. This peculiar effect of H-endorphin on naloxone, together with the non conventional interactions observed in various pharmacological assays, clearly distinguishes H-endorphin from other opioid ligands. A multisite model of the opiate receptor is presented. This model, which is compatible with the well documented evidence for receptors heterogeneity also explains non-conventional interactions between various opioid ligands. PMID- 6288406 TI - A new non-depolarizing myorelaxant with high activity and specificity. AB - The curare-like activity and mode of action of tercuronium (p,p" bistriethylammonium-p-terphenyl dibenzen-sulphonate) have been investigated in experiments with cats, rabbits, mice and pigeons as well as with rat phrenic diaphragm preparations. The curare-like activity of tercuronium was higher than that of tubocurarine and was close to that of pancuronium bromide. The curare like activity of tercuronium was higher than that of tubocurarine and was close to that of pancuronium bromide. The neuromuscular blocking doses (ED95) of tercuronium, (+)-tubocurarine and pancuronium in cats, for example, were 0.08 microM/kg, 0.4 microM/kg and 0.04 microM/kg respectively. The time of development and duration of action were similar to those of (+)-tubocurarine and pancuronium. Tercuronium is a nondepolarizing myorelaxant. In experiments with cats the antagonism of neostigmine (0.1 mg/kg) to tercuronium was more pronounced than in the case of (+)-tubocurarine, pancuronium or gallamine. Tercuronium affected neither the arterial pressure nor the heart rate when given in neuromuscular blocking doses. Tercuronium did not block transmission through autonomic ganglia and had no atropine-like action. Only a 10-fold increase in the dose of tercuronium produced the ganglion blocking effect in cats. Under artificial respiration, cats and rabbits tolerated tercuronium in a dose 200 times exceeding its myoparalytic dose. It is concluded that tercuronium is distinguishable from (+)-tubocurarine by its high neuromuscular blocking activity as well as by its specificity and more pronounced neostigmine antagonism. PMID- 6288407 TI - Concomitant decrease in [3H]imipramine binding in cat brain and platelets after chronic treatment with imipramine. AB - Cats were treated chronically with imipramine (7.5 mg/kg i.p. twice daily for 20 days). Maximal [3H]dihydroalprenolol binding was reduced in the cerebral cortex of the treated animals whereas maximal [3H]spiroperidol binding to 5HT2-receptors was unchanged. Maximal [3H]imipramine binding was decreased to a similar extent in both hypothalamus and platelets of the same animals. This parallel decrease in [3H]imipramine binding in brain and platelets is discussed in relation to the lower [3H]imipramine binding found in platelets from untreated depressed patients as compared to those from control volunteers. PMID- 6288408 TI - Potentiation of the aphrodisiac effect of N-n-propyl-norapomorphine by naloxone. AB - N-n-Propyl-norapomorphine (NPA), a potent dopamine (DA) receptor stimulant, in doses from 0.4 to 80 micrograms/kg i.p. produced a dose-related sexual stimulant effect characterized by recurrent episodes of penile erection (PE). The number of episodes and percentage of responding subjects were proportional to the dose. However, above the maximal effective dose, the effect decreased in a dose-related fashion until beyond 2.5 mg/kg even the natural occurrence of PE was suppressed. Morphine (5 mg/kg), as well as haloperidol (0.3 mg/kg), prevented NPA stimulation. Naloxone, which per se caused a modest increase in PE, markedly potentiated the stimulant effect of low doses of NPA, reversing the inhibitory component of higher doses. We suggest that NPA stimulation of DNA receptors causes release of opiate peptides, dampening the sexual stimulant response. The combination of DA receptor stimulants with naloxone might offer a new possibility for erection defect therapy. PMID- 6288409 TI - Potentiation of phenobarbital-induced anticonvulsant activity by pipecolic acid. AB - Pipecolic acid (PA) is an intermediate of lysine metabolism in the mammalian brain. Recent findings suggest a functional connection of PA as neuromodulator in GABAergic transmission. Since many drugs are postulated to produce their effects by interaction with the central GABA system, the influence of PA on the anticonvulsant activity of phenobarbital was examined. Pretreatment of mice with 50 mg . kg-1 of PA potentiated the suppressing effects of the barbiturate on electrically and chemically induced convulsions. However, there was no potentiation of the behavioral effects and hypothermia induced by phenobarbital. PA itself had no or only little effect on the convulsions, motor function and rectal temperature when given in i.p. doses up to 500 mg . kg-1. Intraventricular administration of 500 microgram of PA also did not suppress either type of convulsion, although it produced ptosis, hypotonia, sedation and hypothermia. The results are discussed in relation to GABA system. PMID- 6288410 TI - Age-dependent decrease of alpha 2-adrenergic receptor number in human platelets. AB - In 36 healthy subjects of various ages (14-76 years) the number of alpha 2 adrenergic receptors in platelets - as determined by [3H]yohimbine binding - and plasma catecholamine levels were measured. A highly significant negative correlation (r = -0.666, P less than 0.001) between the number of alpha 2 adrenergic receptors and age was found; on the contrary, plasma catecholamine concentrations increased with increasing age. Thus, reduced responses in the elderly to adrenergic stimuli may be due to reduced number of adrenergic receptors. PMID- 6288411 TI - Specificity of the beta 2-adrenergic receptor stimulating cyclic AMP accumulation in the intermediate lobe of rat pituitary gland. AB - Changes of cyclic AMP levels were used to assess the specificity of the beta adrenergic receptor in primary cultures of cells prepared from the intermediate lobe of rat pituitary gland. During a 4 min incubation, beta-adrenergic agonists led to a 4 to 6 fold stimulation of cyclic AMP concentration with the following order of potency (Kd values): zinterol (0.75 nM) greater than hydroxybenzylisoproterenol (1.0 nM) greater than (--)-isoproterenol (4.6 nM) greater than soterenol greater than (7.7 nM) greater than (--)-epinephrine (10 nM) greater than OPC 2009 (procaterol, 11 nM) much greater than (--) norepinephrine (300 nM). The potent antagonists cyanopindolol, (--)-propranolol and hydroxybenzylpindolol reversed the stimulatory effect of (--)-isoproterenol at Kd values of 0.4-0.6 nM. Other beta-adrenergic antagonists had the following order of potency: pindolol = (--)-alprenolol = timolol (0.9-1.0 mM) much greater than metoprolol (100 nM) greater than dichloroisoproterenol (300 nM) greater than butoxamine (1100 nM). The beta 1-selective antagonist practolol had a low potency at 700 nM. The stereoselectivity of the receptor is indicated by the 400 to 70 fold higher potency of the (--)-isomers of isoproterenol, epinephrine and propranolol as compared to their (+)-stereoisomers. The data show that the beta adrenergic receptor in the intermediate lobe of the rat pituitary gland is mainly of the bet 2-subtype. Study of this pure population of postsynaptic beta adrenergic receptors where binding could be correlated with other parameters of cellular activity (cyclic AMP formation and alpha-MSH secretion) should yield useful information about the less accessible adrenergic systems of the brain. PMID- 6288412 TI - [3H]diprenorphine receptor binding in vivo and in vitro. AB - In order to investigate opiate receptor binding in vivo, [3H]diprenorphine was given s.c. to rats, and the tracer specifically bound to membraneous high affinity sites was determined with a rapid filtration technique after brain homogenization. Bound [3H]diprenorphine accounted for 70% of the total brain activity after tracer doses. The in vivo binding sites were saturable at 25-30 pmol/g brain. Fifty percent occupancy of the [3H]diprenorphine binding sites in vivo occurred at a dose (10-15 micrograms/kg) that is similar to the antagonistic ED50 of diprenorphine for reversing morphine analgesia. The in vitro binding capacity for [3H]diprenorphine was also approximately 30 pmol/g brain in fresh untreated Tris buffer brain homogenate; however, extensive homogenate dilution or standard membrane washing procedures resulted in a reduction of the [3H]diprenorphine binding site population to 13-22 pmol/g. These results indicate that the opiate receptor system is modified in vitro. Previous studies have shown that the [3H]diprenorphine tracer is retained at cerebral binding sites over several hours in vivo. A diffusion boundary model was proposed to account for the dose dependent tracer retention. In order to investigate the mechanism of the in vivo binding kinetics, [3H]diprenorphine dissociation was measured in brain homogenates after in vivo labeling, immediately following sacrifice of the animals to minimize in vitro artefacts. No differences were found in the dissociation curves at 'infinite' homogenate dilution in the presence or absence of saturating diprenorphine concentrations under various ionic incubation conditions. This result argues against cooperative binding. It is consistent with the hypothesis that the [3H]diprenorphine tracer is retained in vivo because of a diffusion boundary next to the binding sites (receptor micro-compartment) that is destroyed during brain homogenization. PMID- 6288413 TI - Naltrexone-insensitive facilitation and naltrexone-sensitive inhibition of passive avoidance behavior of the ACTH-(4-9) analog (ORG 2766) are located in two different parts of the molecule. AB - Subcutaneous injection of the ACTH-(4-9) analog (OG 2766) in ng amounts prior to the retention test facilitated, while microgram doses attenuated passive avoidance behavior. The inhibitory effect could easily be overcome by treatment with ACTH-(1-10) either before or after ORG 2766 administration. Thus, inhibition of passive avoidance behavior by ORG 2766 probably was not due to competition with ACTH-like peptides or a functional antagonistic influence on brain structures sensitive to ACTH-like peptides. Intracerebroventricular administration of ACTH-(4-10) in a wide dose range (0.5-10.0 micrograms) and of ORG 2766 in low doses (0.5-1.0 ng) facilitated passive avoidance behavior, whereas 'high' doses of ORG 2766 (5.0 and 10.0 ng) and graded doses of COOH terminal tripeptide of ORG 2766 (Phe-D-Lys-Phe; PDLP; 0.5-10.0 ng) attenuated passive avoidance behavior. The NH2 terminal tetrapeptide of ORG 2766 (H-Met/O2/ Glu-His-Phe) facilitated passive avoidance behavior, whereas the NH2 terminal tripeptide (H-Met/O2/-Glu-His) was ineffective. Naltrexone pretreatment antagonized the attenuating effect of ORG 2766 and PDLP. Following pretreatment with this opiate antagonist both 'low' and 'high' doses of ORG 2766 and the NH2 terminal tetrapeptide of ORG 2766 induced facilitation of passive avoidance behavior, while PDLP was ineffective in the presence of naltrexone. Thus, ORG 2766 exerts a dual effect on passive avoidance behavior. The facilitating effect of ORG 2766 resides in the NH2 terminal part and is unrelated to naltrexone sensitive brain opiate receptor sites, whereas the inhibiting influence is located in the COOH terminal part of the peptide and depends on naltrexone sensitive brain opiate receptor sites. PMID- 6288414 TI - Cardiovascular effects of beta-endorphin after microinjection into the nucleus tractus solitarii of the anaesthetised rat. AB - The cardiovascular effects of beta-endorphin after administration directly into the nucleus tractus solitari (NTS) of urethane-anaesthetised rats were investigated. Unilateral injection resulted in a U-shaped dose-response relationship with a fall in mean arterial pressure and heart rate occurring at low doses (less than 10 ng). No change in respiratory frequency was observed at any of the doses examined. The hypotensive effects of beta-endorphin were anatomically specific and restricted to the NTS. The depressor response was prevented and bradycardia reduced by naloxone (1 mg/kg s.c. or 10 ng injected into the NTS) and also by beta-endorphin antiserum (1:50 dilution) but not by antiserum to [Met5]enkephalin (1:50 dilution) applied locally into the NTS. The beta-endorphin antiserum caused a rise in blood pressure when administered alone. Conversely microinjection of antiserum to [Met 5]enkephalin resulted in a brief depressor response. Doses of beta-endorphin larger than 10 ng induced a rise in blood pressure accompanied by variable effects on heart rate. Similarly unilateral administration of Des-tyr-endorphin (100 pg) resulted in a blood pressure increase and [D-Ala2,Met5]enkephalin produced a dose-related pressor response and tachycardia. The results indicate that at least two separate endorphin systems are involved in cardiovascular control at the level of NTS, one being depressor in nature (beta-endorphin-like) and the other pressor ([met5]enkephalin-like). PMID- 6288415 TI - Comparison of angiotensin converting enzyme inhibitors captopril and MK421-diacid in guinea pig atria. AB - Angiotensin I (AI) and angiotensin II (AII) caused concentration-dependent increases in atrial rate in guinea pig isolated right atria. Converting enzyme inhibitors captopril and MK421-diacid did not alter the responses to AII but displaced the curves to AI to the right. The atrial response to generated AII from AI was used as a bioassay to estimate the dissociation constants of converting enzyme inhibitors (Kb) and test for kinetics of simple competition. MK421-diacid was 12-40 times more potent than captopril. However, estimations of Kb for captopril and MK421-diacid were unsatisfactory because at high concentrations of inhibitors the curves to AI were not displaced according to simple competition. We conclude that AI in high concentration can stimulate AII receptors accounting for the stationary displacement of curves to AI in the presence of converting enzyme inhibitors. MK421-diacid also potentiates responses to bradykinin in this assay. PMID- 6288416 TI - Role of vasopressin in the ACTH response to isoprenaline. AB - The present study investigated whether or not the beta-sympathomimetic amine isoprenaline, given systemically to conscious rats, influences corticotrophin (ACTH) release and if so, what could be the role of vasopressin in this response. Isoprenaline (i.m.) elevated plasma ACTH-like immunoreactivity (ACTHi) in a time- and dose-dependent manner. The highest dose of isoprenaline used (240 microgram/kg) raised plasma ACTHi about six fold. Most of the ACTHi co-migrated with porcine ACTH-(1-39) on Sephadex G-50 column chromatography. The beta receptor antagonist propranolol abolished the increase in plasma ACTHi induced by isoprenaline, as did dexamethasone pretreatment. The increase in plasma ACTHi following isoprenaline (120 microgram/kg) injection was diminished by about 35% in rats congenitally lacking vasopressin (Brattleboro rats), when compared to normal rats. The vasopressin analogue, [1-deaminopenicillamine, 2-(O methyl)tyrosine]-arginine-vasopressin, almost completely prevented the rise in plasma ACTHi provoked by i.v. injection of arginine vasopressin and diminished by about 40% the isoprenaline-(120 microgram/kg)-caused ACTHi release. However, this vasopressin analogue had no effect in Brattleboro rats. These results indicate that isoprenaline, given systemically, stimulates the release of pituitary ACTH and this response appears to be mediated in part by vasopressin acting as an ACTH releasing factor. PMID- 6288417 TI - Evidence for separate receptors for ATP and adenosine in the guinea-pig taenia coli. AB - Inhibitory actions of three pairs of congeneric ATP and adenosine analogues on the isolated guinea-pig taenia coli were compared with the actions of ATP and adenosine. 8-Bromoadenosine, and 9-beta-D-arabinofuranosyladenosine (ara adenosine) were inactive; 2'-deoxyadenosine was a weak partial agonist. 8-Bromo ATP, ara-ATP and 2'-deoxy-ATP behaved like ATP and elicited rapid relaxations of the taenia but were not potentiated by dipyridamole. The divergent effects of identical structural modifications to ATP and adenosine provide evidence for separate adenosine and ATP receptors in the taenia coli. PMID- 6288419 TI - Transferrin binding to K562 cell line. Effect of heme and sodium butyrate induction. PMID- 6288420 TI - Distribution of actin, myosin, actin-binding protein and gelsolin in cultured lymphoid cells. PMID- 6288418 TI - Xanthine derivatives as adenosine receptor antagonists. AB - The potency of a series of xanthine derivatives as adenosine antagonists was studied in fat cells (A1-receptors) and hippocampal slices (A2-receptors) and on L-[3H]phenylisopropyladenosine (PIA) binding in membranes from rat cortex. The order of potency in all three tests systems was: diethyl-8-phenyl-theophylline greater than 8-phenyltheophylline greater than 8-p-sulfophenyltheophylline greater than verrophylline greater than isobutylmethylxanthine greater than theophylline caffeine greater than theobromine. Enprofylline was about 20 times more potent in the hippocampus system than in the other two systems. PMID- 6288421 TI - Experimental reversal of polarity in chick embryo epiblast sheets in vitro. PMID- 6288422 TI - Replication and amplification of recombinant plasmid molecules as extra chromosomal elements in transformed mammalian cells. PMID- 6288424 TI - The non-selective junctional communication phenotype of normal and transformed human epidermal keratinocytes in vitro. PMID- 6288423 TI - Inhibition of dihydrofolate reductase gene expression following serum withdrawal or db-cAMP addition in methotrexate-resistant mouse fibroblasts. PMID- 6288425 TI - A spin label study of human lens membranes. PMID- 6288426 TI - Relationship between angiotensin-converting enzyme and lysozyme in sarcoidosis. AB - The relationship between serum angiotensin-converting enzyme (SACE) and serum lysozyme (SLZ) was examined in 48 patients with sarcoidosis. The level of at least one enzyme was elevated in all 41 cases with active sarcoidosis; both enzymes were elevated in 24. SACE and SLZ were positively correlated. The elevation of SLZ or SLZ+SACE was most frequent in patients with pulmonary parenchymal involvement or extrathoracic manifestations. Both enzymes are suitable for disease monitoring, but in most cases the more specific SACE must be preferred as a measure of disease activity, although SLZ seems to be a little more sensitive. Blood for SACE analysis must arrive at the laboratory not later than 3 days after blood collection in order to avoid enzyme inactivation. PMID- 6288428 TI - cAMP concentration in the rat's preoptic region and cerebral cortex during sleep deprivation and recovery induced by ambient temperature. AB - cAMP concentrations in the preoptic region and cerebral cortex were studied in rats during exposure to low ambient temperature (-10 degrees C) and after return to control ambient temperature (22 degrees C). Significant changes in cAMP concentration were found only in the preoptic region. On prolonged exposure to low ambient temperature the nucleotide concentration decreased and the circadian rhythm, observed in control conditions, disappeared. Return to control ambient temperature after exposure to low ambient temperature induced a steep increase and a long-lasting plateau in cAMP concentration. The results are discussed in terms of interaction between thermoregulatory and sleep-wakefulness processes. PMID- 6288427 TI - Neuronal interactions in the substantia nigra pars reticulata through axon collaterals of the projection neurons. An electrophysiological and morphological study. AB - Substantia nigra pars reticulata (SNr) neurons, antidromically activated following stimulation of the dorsal thalamus and/or superior colliculus were intracellularly stained with HRP. Light microscopic analysis revealed that the labeled SNr neurons have axon collaterals arborizing within SNr. Axon collaterals of SNr neurons partially overlapped with the dendritic fields of their parent cells and also extended beyond the parent dendritic fields. The labeled axon terminals did not closely appose the parent cell processes, suggesting that the collaterals most likely terminate on neurons other than the parent cell. Electrical stimulation of either the thalamus or the superior colliculus induced monosynaptic and polysynaptic IPSPs in SNr cells. The polysynaptic IPSPs evoked from thalamic stimulation disappeared following hemitransection of the brain just rostral to the thalamus while the monosynaptic IPSPs remained the same. Since there are no known afferents from either thalamus or superior colliculus to SNr, we consider that these monosynaptic IPSPs are due to activation of the recurrent collaterals of SNr projection cells. The results of this study indicate that projection neurons of SNr also have an inhibitory role within the SNr. PMID- 6288429 TI - Vestibular signals in the posterior vermis of the alert monkey cerebellum. AB - Head velocity-related modulations in Purkinje cell activity were observed in lobules VI and VII of the cerebellar vermis. The sensitivities and phase shifts with respect to head velocity of Purkinje cells were comparable with vestibular responses observed in the fastigial nuclei. The results support the hypothesis of a vermal target velocity correlate and necessitate the inclusion of vermal lobules VI and VII among the cerebellar regions involved with vestibular information processing. PMID- 6288430 TI - Electrophysiological evidence against the hypothesis that corticospinal fibres send collaterals to the lateral reticular nucleus. AB - Extracellular focal synaptic potentials (FSPs) were recorded in forelimb segments of cats in the region where corticospinal volleys (pyramidal stimulation) gave a maximal FSP. No FSP was recorded in this region on stimulation of the medial part of the lateral reticular nucleus (LRN), provided that ventral spinal pathways through which FSPs can be evoked from the LRN region were transected. It is postulated that the projection from the cerebral cortex to the LRN is not by collaterals from corticospinal neurones but rather by separate corticofugal neurones. PMID- 6288431 TI - Effect of sodium ions on penicillin-induced epileptiform activity in vitro. AB - Intra- and extracellular recordings were obtained from the CA1 region of guinea pig hippocampal slices maintained in vitro. We studied the effect of reducing the extracellular sodium concentration on penicillin-induced epileptiform responses. In control experiments, Tris and choline were assayed as sodium substitutes. Choline was found unsuitable, since it induced repetitive firing in the absence of any convulsant agent. Replacement of 50% of the extracellular sodium ( [Na+]o) with Tris reduced the amplitude of the presynaptic fiber volley, the field EPSP, and the population spike. Intracellular studies showed that when [Na+]o was lowered, action-potential amplitudes were reversibly depressed by an amount close to that predicted by the Nernst relation. Orthodromically elicited epileptiform discharges, induced by penicillin, were reduced in a low-sodium medium when constant stimulus currents were employed. If orthodromic stimulus strengths in normal and low-sodium states were equated on the basis of the field-EPSP amplitude, no significant diminution of the depolarizing-wave component of the epileptiform response was observed. These results suggest that a synaptic component underlies penicillin-induced epileptiform discharges. PMID- 6288435 TI - An electrophysiological analysis of chronic ethanol neurotoxicity in the dentate gyrus: distribution of entorhinal afferents. AB - This study investigated the persistent effects of chronic ethanol consumption on the distribution of entorhinal afferents to stratum moleculare of the dentate gyrus. Rats were maintained on ethanol, or sucrose-containing liquid diets, for a period of 20 weeks but were withdrawn from the special diet for at least 8 weeks prior to acute electrophysiologic recordings. One-dimensional laminar analyses were obtained by stepping a microelectrode in 25 microns increments across both the dorsal and ventral blades of the dentate gyrus and sampling the field potentials at each point. Current-source density (CSD) was calculated from the field potential data. Electrical stimulation of the angular bundle elicited a short-latency, negative field potential covering the outer 2/3 of the molecular layer. CSD analysis revealed a major current sink in stratum moleculare bounded by a major current source, localized to the hilus, granule cell layer, and proximal stratum moleculare, and a minor current source localized to the outer molecular layer. Chronic ethanol treatment resulted in (1) a significant shrinkage of the spatial extent of the current sink in stratum moleculare, (2) a significant reduction in the distance from the peak inward synaptic current to the granule cell layer and (3) no change in the distance from the proximal inversion point to the granule cell layer. Taken together, these results indicate a loss of entorhinal afferents in the outer molecular layer. Coupled with available anatomical evidence, these results suggest that chronic ethanol treatment produces a preferential loss of lateral entorhinal afferents to the dentate gyrus. PMID- 6288434 TI - The geniculocortical system in the early postnatal kitten: an electrophysiological investigation. AB - The transmission of neuronal impulses in the central visual system in kitten 7-42 days old has been investigated using electrophysiologic techniques. It was found that shortly after birth the axonal conduction velocities in the geniculocortical, in the corticogeniculate and in the commissural pathways of areas 17, 18, and 19 are less than 1.2 m/s, increasing up to maximal 10 m/s at the end of the first postnatal month. The monosynaptic delays across geniculocortical and commissural synapses in area 17 as measured during the third and fourth postnatal week are extremely long (between 2.8 and 6.1 ms) and do not change significantly during this time. During the first postnatal month, visual cortical neurons fatigue rapidly to visual and electrical stimulation; also, the proportion of neurons receiving polysynaptic inputs is much smaller than in the adult animal. Accordingly, neurons having complex receptive fields are rarely found in area 17 during the first postnatal month. Our experiments show that the geniculocortical and also the intracortical transmission of neuronal impulses are still immature at the end of the first postnatal month; at this time the systems for orientation and direction selectivity in the visual cortex are almost fully developed. PMID- 6288432 TI - Relationship between EPSP shape and cross-correlation profile explored by computer simulation for studies on human motoneurons. AB - We used a computer model (making several simplifying assumptions) to explore the relationship between the characteristics of an excitatory postsynaptic potential (EPSP) and the profile of the change in firing probability that occurred when that EPSP was delivered to a rhythmically discharging neuron. In circumstances applicable to studies on human motoneurons we found that the magnitude of the period of increased firing probability in the cross-correlation (produced by the rising phase of the EPSP) was related to the number of stimuli and to the proportion of the interspike interval that the EPSP was within reach of threshold. The interstimulus interval and the statistical distribution of the motoneuron interspike intervals were of little consequence. For this model the subsequent period of reduced firing probability was proportional to the amplitude of the EPSP and not to the duration of its falling phase. PMID- 6288433 TI - Characteristics of postsynaptic potentials produced in single human motoneurons by homonymous group 1 volleys. AB - Changes in the firing probability of voluntarily activated human tibialis anterior motor units occurred when the muscle nerve was stimulated below the threshold of motoneuron axons. A prominent period of increased firing probability was considered to result from the group 1 composite EPSP. The rise time of this composite EPSP, estimated from the duration of the period of increased firing probability, was between 2 and 5 ms. One estimate of the relative amplitude of the EPSP was derived from the cross-correlation and tested by inserting synthetic EPSPs into a computer simulation of a rhythmically discharging neuron. A second estimate of the amplitude was obtained by delivering stimuli with various delays within the interspike interval to determine the position (as a percentage of the mean interspike interval) at which the postsynaptic potential was capable of bringing the membrane potential to threshold. The two estimates were in reasonable agreement. The largest of these EPSPs could bring the motoneuron to threshold after 39% of the elapsed interspike interval. The falling phase of the postsynaptic potential was explored by delivering double stimuli within the interspike interval to produce temporal summation. The duration of the falling phase, so tested, was between 5 and 20 ms. An attempt was made to deduce the general form of the trajectory representing the effective distance of an EPSP from threshold during the interspike interval from the alterations in the cross correlation profile that occurred when motoneuron firing rate was voluntarily altered. PMID- 6288437 TI - Membrane desmosterol and the kinetics of the sarcolemmal Na+,K+-ATPase in myotonia induced by 20,25-diazacholesterol. PMID- 6288436 TI - Treating monocularly deprived lambs with 4-aminopyridine produces rapid changes in ocular dominance only after short periods of deprivation. AB - Lambs of various ages (4-30 days) were monocularly deprived for periods ranging from 4 to 51 days after which they were prepared for conventional electrophysiological recording from the striate cortex. Sufficient units were sampled to obtain a representative ocular dominance (OD) histogram. A second sample of units was then obtained after the lambs had been given an i.v. dose of 4-aminopyridine (4-AP; 0.5-5.3 mg/kg) which increases synaptic transmission. For four out of five lambs in which the deprivation had been of 4-5 days duration there was a significant increase in responsiveness of the deprived eye to stimulation after the 4-AP had been given. By contrast, only one out of four lambs which had been deprived for 19-51 days showed a significant recovery after 4-AP treatment. The results suggest that during the initial stages of monocular deprivation the deprived eye remains connected to, but is less effective in driving, cortical cells. One explanation of the failure to reactivate the deprived eye after long periods of deprivation is that the deprived eye becomes anatomically disconnected from cortical cells. PMID- 6288438 TI - A biochemical investigation of spinal cord after contusive injury. PMID- 6288439 TI - Lasting influence of amygdaloid kindling on cholinergic neurotransmission. PMID- 6288440 TI - The in vivo effect of asbestos on the SiO2-induced macrophage fibrogenic factor and alkaline ribonuclease. PMID- 6288441 TI - Storage of the complement components C4, C3, and C 3-activator in the human liver as PAS-negative globular hyaline bodies. AB - Liver biopsies of a 58-year-old clinically healthy patient with a hepatomegaly and intracisternal PAS-negative globular hyaline bodies were immunofluorescent optically examined for the content of the complement components C 1 q, C 4, C 9, C 1-inactivator, C 3-activator. Further examinations were performed for fibrinogen, IgG, IgA, IgM, IgD, IgE, L-chain (type chi and lambda), alpha 1 antitrypsin, alpha 1-fetoprotein, alpha 1- and alpha 2-glycoprotein, cholinesterase, ceruloplasmin, myoglobin, hemopexin, HBsAg and HBsAg. Th inclusion bodies reacted with antisera against the complement components C 4, C 3 and C 3-activator, as also identified by double immunofluorescence. Probably this is a disturbance of the protein metabolism of the liver cell with abnormal complement storage in the presence of normal total complement and normal complement components in the serum. PMID- 6288442 TI - Fluoxymesterone-induced inclusion bodies in dog liver. PMID- 6288444 TI - Effects of cholecystokinin octapeptide and hydrocortisone on the exocrine pancreas of fed and fasted 24-hour-old rats: an electron microscopic study. PMID- 6288443 TI - Crinophagic inclusions in gastric chief cells of mice with Chediak-Higashi syndrome. PMID- 6288446 TI - Enzymatic transfer of ATP gamma S thiophosphate onto the 5'-hydroxyl of an oligonucleotide as a route to reactive oligonucleotide derivatives. PMID- 6288445 TI - Angiotensin II inhibits the accumulation of cyclic AMP produced by glucagon but not its metabolic effects. PMID- 6288447 TI - Arrangement of the ribosomal RNA genes in Schizosaccharomyces pombe. PMID- 6288448 TI - The thermodependence of the activity of integral enzymes in liver plasma membranes: evidence consistent with a functionally asymmetric lipid bilayer. PMID- 6288449 TI - Peptide-receptor protein relationships: importance in estradiol feedback. PMID- 6288450 TI - Complex formation between metarhodopsin II and GTP-binding protein in bovine photoreceptor membranes leads to a shift of the photoproduct equilibrium. PMID- 6288451 TI - Tumor promoter teleocidin inhibits internalization and nuclear accumulation of epidermal growth factor in cultured human hepatoma cells. PMID- 6288452 TI - Inhibition of calmodulin-stimulated cyclic nucleotide phosphodiesterase by the insecticide DDT. PMID- 6288453 TI - Circadian variations of A-mediated transport in rat-liver plasma membrane vesicles. PMID- 6288454 TI - Immunoautoradiographic detection of epidermal growth factor receptors after electrophoretic transfer from gels to diazo-paper. PMID- 6288455 TI - Antagonistic effects of insulin on the corticosterone-induced increase of phosphatidate phosphohydrolase activity in isolated rat hepatocytes. PMID- 6288456 TI - Polymyxin B induces interdigitation in dipalmitoylphosphatidylglycerol lamellar phase with stiff hydrocarbon chains. PMID- 6288457 TI - A cyclic AMP-medicated intersubunit disulfide crosslinking reaction of the regulatory subunit of type II cyclic AMP-dependent protein kinase. PMID- 6288458 TI - Phosphorylation of human fibrinogen in vitro with calcium-activated phospholipid dependent protein kinase and [32P]ATP. PMID- 6288459 TI - Hydroxyapatite chromatography of the D-glucose transport protein of human erythrocyte membranes. PMID- 6288461 TI - Binding of the intramitochondrial ADP and its relationship to adenine nucleotide translocation. PMID- 6288460 TI - Scope of the ATP--ubiquitin system for intracellular protein degradation. PMID- 6288463 TI - 6-beta-(Trifluoromethane sulfonyl)-amido-penicillanic acid sulfone: a potent inhibitor for beta-lactamases. PMID- 6288462 TI - Conjugation of specific antibodies to Sendai virus particles: a new tool for targeting of fusogenic vesicles. PMID- 6288464 TI - Synthesis of 6'-O-p-azidobenzoylatractyloside, a short arm photoactivable derivative of atractyloside: studies of its binding and inhibitory properties. PMID- 6288465 TI - The quaternary structure of phosphoglycerate mutase from yeast: evidence against dissociation of the tetrameric enzyme at low concentrations. PMID- 6288466 TI - AhaIII: a restriction endonuclease with a recognition sequence containing only A:T basepairs. PMID- 6288467 TI - Strychnine and local anesthetics block ion channels activated by veratridine in neuroblastoma x glioma hybrid cells. PMID- 6288468 TI - The alpha 2 HS glycoprotein receptor on lymphocytes transformed by Epstein--Barr virus. AB - The alpha 2HS glycoprotein receptor from lymphocytes transformed by Epstein--Barr Virus was isolated by affinity chromatography. The protein receptor has a monomer Mr of 48 000, which is similar to the Epstein--Barr virus-determined nuclear antigen (EBNA), and pI = 7.2. Like EBNA the 48 000 Mr component in unfractionated labelled detergent solubilised cell supernatants also binds DNA. These results may suggest some similarity between the alpha 2HS receptor and EBNA. PMID- 6288469 TI - Critical evaluation of the proton-translocating property of cytochrome oxidase in rat liver mitochondria. PMID- 6288470 TI - The intrinsic substrate specificity of a cyclic nucleotide-independent protein (histone) kinase. PMID- 6288471 TI - Internal sequence repeats and the path of polypeptide in mitochondrial ADP/ATP translocase. PMID- 6288472 TI - Cyclic nucleotide-dependent inactivation of yeast fructose 1,6-bisphosphatase by ATP. PMID- 6288473 TI - In vitro assembly of cytochrome oxidase from separately accumulated subunits: a reconsideration. PMID- 6288474 TI - Mapping of the cytochrome c binding site on cytochrome c oxidase. PMID- 6288475 TI - Subunit structure and dynamics of the insulin receptor. AB - A model for the minimum subunit composition and stiochiometry of the physiologically relevant insulin receptor has been deduced based on results obtained by affinity labeling of this receptor in a variety of cell types and species. We propose that the receptor is a symmetrical disulfide-linked heterotetramer composed of two alpha (apparent Mr = 125,000) and two beta (apparent Mr = 90,000) glycoprotein subunits in the configuration (beta-S-S alpha)-S-S-(alpha-S-S-beta). The disulfide or disulfides linking the two (alpha-S S-beta) halves (class I disulfides) exhibit greater sensitivity to reduction by exogenous reductants than those linking the alpha and beta subunits (class II disulfides). When the class I disulfides are reduced by addition of diothiothreitol to intact cells, the receptor retains its ability to bind insulin and to effect a biological response. The beta subunit contains a site at about the center of its amino acid sequence that is extremely sensitive to proteolytic cleavage by elastaselike proteases, yielding a beta 1 fragment (Mr = 45,000) that remains disulfide linked to the receptor complex and a free beta 2 fragment. Binding of insulin to the receptor complex appears to result in the formation or stabilization of a new receptor conformation as evidenced by an altered susceptibility of the alpha subunit to exogenous trypsin. A receptor structure with high affinity for insulinlike growth factor (IGF) I and low affinity for insulin in fibroblast and placental membranes has also been affinity labeled. It exhibits the same structural features found for the insulin receptor, including two classes of disulfide bridges and beta subunits highly sensitive to proteolytic cleavage. These recent observations identifying the presence of distinct insulin and IGF-I receptors that share similar complex structures suggest that these hormones may also share common mechanisms of transmembrane signaling. PMID- 6288477 TI - Chemical mediator or mediators of insulin action: response to insulin and mode of action. AB - Studies with a subcellular system demonstrated that the interaction of insulin with the adipocyte plasma membrane resulted in the generation from the plasma membrane of a mediator that activated mitochondrial pyruvate dehydrogenase (EC 1.2.4.1). The insulin-sensitive chemical mediator from the plasma membrane has been partially characterized. It has a molecular weight of 1000-1500. The chemical mediator has been extracted from skeletal muscle, adipocytes, hepatoma cells, and IM-9 lymphocytes. Insulin increased the amount or activity of the mediator in the first three cell types, whereas insulin decreased the activity or amount of the mediator in IM-9 lymphocytes. These insulin-induced variations were consistent with the biological responses of these cells to insulin treatment. The activities of insulin-sensitive enzymes, including pyruvate dehydrogenase, adipocyte low Km 3':5'-cyclic-AMP phosphodiesterase (EC 3.1.4.17), and adipocyte plasma membrane [Ca2+ + Mg2+]-ATPase were shown to be altered by the chemical mediator. The mediator may act by altering various protein kinases and phosphoprotein phosphatases that modulate the state of phosphorylation and activity of these enzyme systems. The existence of two mediators is proposed. The first may mediate dephosphorylation of various substrates, and the second may influence phosphorylation. PMID- 6288476 TI - A proteolytic mechanism for the action of insulin via oligopeptide mediator formation. AB - Evidence is presented that the chemical mediator of insulin action is a peptide(s) and most likely glycopeptide(s). The mediator is formed proteolytically because 1) protease inhibitors inhibit insulin action and 2) trypsin mimicks insulin action via mediator formation. Trypsin mediator does not faithfully reproduce the action of insulin mediator, which indicates that the sites of proteolytic cleavage by insulin and trypsin differ. A coordinated multivalent proteolytic mechanism by which insulin acts to trigger an external membrane-bound protease to cleave mediator from a membrane glycoprotein precursor is presented. PMID- 6288478 TI - Anion and proton transport in chromaffin granules. AB - Both intact chromaffin granules and ghosts behave as osmometers in response to the relative internal and external concentrations of solutes. The permeability properties of the granule membrane can therefore be studied by measuring the osmotic consequences of ion permeation. These include lysis of intact granules and shrinking or swelling of ghosts. This approach has been used to identify at least two types of selective anion transport sites in the membrane. One is observed in the presence of K+ ionophores and appears to be electrogenic in nature. It is insensitive to a group of compounds known to inhibit anion transport in erythrocytes. The other site is sensitive to those inhibitors and is involved with electroneutral cotransport with protons. This proton entry can be driven by either the proton-pumping ATPase of the granule membrane or by a simple inward concentration gradient of protons. The proton entry-coupled anion transport site may be structurally related to the ATPase itself based on parallel anion and inhibitor sensitivities of the site and the enzyme. One or both of the anion transport sites described here may play a role in regulating exocytosis from cells based on a chemiosmotic mechanism that has been postulated to be responsible for this process. PMID- 6288479 TI - Proton-translocating ATPase of chromaffin granule membranes. AB - Active transport of catecholamines into chromaffin granules is driven by the transmembrane pH gradient and membrane potential, created by an electrogenic proton-translocating ATPase in the granule membrane. The ATPase activity of highly purified chromaffin granule membranes is inhibited by a number of agents in common with mitochondrial ATPase, and also by antibodies raised against mitochondrial F1. Dichloromethane treatment of these membranes solubilizes an enzyme that is closely similar to mitochondrial F1, but distinguishable from it by its interaction with specific antisera and the inhibitor aurovertin. Chromaffin granule membranes contain a low-molecular-weight protein that reacts with dicyclohexylcarbodiimide; it can be extracted into chloroform-methanol, and is of higher electrophoretic mobility than the corresponding mitochondrial protein. Evidence is presented that this is a component of the proton translocating ATPase complex. PMID- 6288480 TI - Dietary fiber, lipid metabolism, and atherosclerosis. AB - Despite the physiochemical complexity of dietary fibers (plant cell walls) and their individual components, there is substantial epidemiologic, clinical, and experimental evidence that these dietary components may have a role in modifying certain risk factors in coronary heart disease. Particulate fibers, such as wheat bran, do not appear to significantly alter plasma lipids or lipoprotein distributions in humans, or the atherogenicity of diets in experimental animals. Dietary fibers found in fruits, legumes, and vegetables, in contrast, show more definitive responses. Among the fiber isolates, the gelling and mucilaginous fibers, such as pectins and guar gum, predictably decrease circulating lipids in humans and animals and increase excretion of fecal metabolites of cholesterol, the bile acids. These fibers and fiber components can be shown to influence luminal solubility of lipids and the extent of lymphatic absorption of both cholesterol and triglyceride. In addition, these same fibers are effective in reducing postprandial levels of glucose, insulin, and other hormones. These direct effects on lipid absorption, and secondary effects of glucose and insulin on hepatic and peripheral lipoprotein metabolism, can account for many of the hypolipidemic responses to specific dietary fibers or their components, and may be of long-term consequence in coronary heart disease. PMID- 6288481 TI - Evidence for the presence of follicle-stimulating hormone receptor antibody in human serum. AB - A male patient presented with polyostotic fibrous dysplasia, elevated serum gonadotropin levels, primary gonadal failure, and an immunoglobulin M (IgM) monoclonal gammopathy. When the patient's serum was added to radioiodinated human follicle-stimulating hormone (125I-hFSH)-bovine testis membrane-receptor complex solubilized by detergent, followed by the addition of antihuman immunoglobulin G ((anti-hIgG), the preformed complex was precipitated. No such precipitation was seen when normal human or rabbit serum, serum from a patient with polyostotic fibrous dysplasia, or serum from other patients with polyclonal gammopathies were utilized. The patient's serum did not precipitate free 125I-hFSH. Detergent solubilized testis receptor not complexed to hFSH, as well as highly purified radioiodinated receptor were also precipitated when this serum was added followed by anti-hIgG. Our results indicate that serum from the patient contained antibodies to FSH testis receptor. It is possible that antibodies to gonadal receptors may exist in other patients, although it is not yet possible to assume a causal relationship between the presence of receptor antibodies and gonadal failure. PMID- 6288482 TI - [Effect of immunosuppression with anti-cytochrome oxidase serum on the functional activity of peripheral blood and lymph node lymphocytes following skin allotransplantation]. PMID- 6288483 TI - [Concentrations of cAMP and cGMP during adaptation of brain tissue to ischemia]. PMID- 6288484 TI - [Regulation of calcium metabolism in the cells of different portions of the heart of a warmblooded animal]. PMID- 6288485 TI - [Effect of preparations altering cAMP metabolism on the bioelectrogenesis of the skeletal muscles]. AB - In the frog isolated m. sartorius, microelectrode technique and in vivo TV microscopy differentiating bioelectrical phenomena in separate fibers of different groups (dark, light, intermediate), revealed that the drugs altering cAMF exchange (adrenaline, caffeine, imidazole) and used in concentration 10(-5) M, affect to different extents the resting potential and the AP. The effect of the drugs is more obvious in respect to excitatory potential. Mechanism of this phenomenon is discussed. PMID- 6288486 TI - [Clinical study on the angiotensin I-converting enzyme in human urine. (I) Partial purification, enzymic characteristics and excretion in normal subjects]. AB - The angiotensin I-converting enzyme in normal human urine was partially purified with ammonium surfate (3.2M), DEAE-Cellulose ion exchange chromatography (0-0.5M NaCl gradient), and Sephadex G 200 gel filtration. The enzyme was separated into three forms which had different molecular weights of 700000, 290000 and 40000, respectively. The enzymic biochemical characteristics of these three enzymes, however, were identical with regard to their inhibitory effects (bradykinin potentiator c, arg-pro-pro, o-phenanthroline and EDTA), Cl- dependency, optimal pH (8.3) and temperature (37 degrees C), and Km value (2.3mM). The enzymic activity was determined in five normal subjects in three conditions of dietary sodium intake (51, 153 and 340mEq for 5 days, respectively). The enzymic activity correlated well with the concentration of the excreted sodium (r = 0.87, p less than 0.001). There was no significant relation between the enzymic excretion and the concentration of the excreted potassium, nor between the activity and the creatinine excretion. It is suggested that the origin of urinary angiotensin I converting enzyme is the kidney, and that the enzyme might regulate sodium excretion in cooperation with renal kallikrein-kinin system. PMID- 6288487 TI - [Calcium ions in the steroidogenic action of HCG]. AB - Extensive studies have recently been conducted on the expression of hormone actions, especially on its mechanisms. Sutherland, et al. identified the function of C-AMP as an intracellular messenger of peptide hormones. Recent studies by several investigators including ours have concluded that the role of C-GMP and calcium are as important as that of C-AMP. In this report we present out findings on the role of Ca ions in the acceleration of steroid production in isolated porcine cells of the corpora lutea. (1) The incubation time needed to compare the production of progesterone and that of C-AMP is at least 120 mins. (2) Production rates of both C-AMP and progesterone in the presence of HCG were maximum at 10( 7) g/ml HCG. Inclusion of Ca ions in the medium increases this production rate. (3) When Ca ions are included in the medium, the accelerated rate of progesterone production takes on its highest value at 1.2mM Ca. (4) Verapamil and cycloheximide inhibit the production of progesterone. (5) The binding capacity of 125I-HCG combined with corpora lutea cells is independent of the Ca concentration in the medium. (6) Ca uptake into corpora lutea cells increases according to the concentration of HCG until it reaches a plateau at 10(-7) g/ml HCG. This acceleration by HCG is affected by Ca and is remarkably inhibited by verapamil. These results indicate the important role of Ca ions in the acceleration of progesterone production compared to that of C-AMP. PMID- 6288488 TI - [Crystalline quartz--one cause of progressive scleroderma? Epidemiologic studies on the simultaneous appearance of progressive scleroderma and silicosis]. PMID- 6288489 TI - Occurrence of phosphodiesterase IV in the developing human brain, liver and placenta. AB - The presence of phosphodiesterase IV has been demonstrated in the human fetal brain, liver and placenta as early as in the 6th week of intrauterine development. The enzyme activity in each tissue increases with gestation, being maximum at 18-21 wk and then decreases. The Km values of this enzyme for bis-(p nitrophenyl)-phosphate hydrolysis in the brain, liver and placenta are 2.94 mM, 1.47 mM and 1.66 mM, respectively. Presence of sulfhydryl group in the active center of the placental enzyme has been demonstrated with the help of cationic study. EDTA inhibits the enzyme in all three tissues. Effect of concanavalin A reveals the absence or unexposition of glucose, mannose and N-acetylglucosamine moieties in the active site of the enzyme in each of the three tissues. Maximum enzyme activity has been found to be localized in the soluble supernatant fraction obtained on centrifuging the brain and liver homogenate at 105,000 x g and in 20,000 x g pellet of the placenta. PMID- 6288490 TI - Short-term effect of glucagon on gluconeogenesis and pyruvate kinase in rabbit hepatocytes. PMID- 6288491 TI - Phosphorylation of purified mitochondrial cytochromes P-450 (cholesterol desmolase and 11 beta-hydroxylase) from bovine adrenal cortex. AB - Two key steroidogenic mitochondrial cytochromes P-450 (cholesterol side-chain cleavage (scc) and 11 beta-hydroxylation (11 beta)) were purified from bovine adrenal cortex and examined as potential phosphorylatable substrates using purified cAMP-dependent protein kinase subunit (C) and A type (CKA) and G type (CKG) cAMP-independent casein kinases. Of the two cytochromes P-450, only P-450 11 beta was able to incorporate phosphate from ATP in the presence of C (Km = 7.5 microM), whereas CKA and CKG were ineffective. Phosphorylation of P-450 11 beta (maximum incorporation of 1 mole of 32P per mole of cytochrome, only on serine residues) did not modify the enzymatic activity of an 11 beta-hydroxylation system reconstituted in vitro from purified components, when adrenodoxin was in excess in the reaction. However, kinetic studies showed that P-450 11 beta phosphorylation strikingly increases the P-450 11 beta-adrenodoxin affinity in a phosphorylation-dependent manner. This would result in a net increase in 11 beta hydroxylase activity under in vivo conditions where adrenodoxin availability is limited. Possible significance of these observations in the regulation of differentiated adrenocortical functions remains to be further examined. PMID- 6288492 TI - ACTH-induced change of cAMP-dependent protein-kinase repartition in bovine adrenal cells. AB - Acute ACTH stimulation of isolated adrenal cells produced a modification of the subcellular distribution of cAMP-dependent protein kinase. Within 20 min, the protein-kinase ratio in all subcellular fractions, particularly in the 175 000 x g supernatant, was increased. Total protein-kinase activity, as well as the specific activity of both the homogenate and particulate enzymatic activities, was decreased while that of the 175 000 x g supernatant was increased. However, the increase of the soluble kinase activity represented only 29-46% of the lost particulate activity. On the other hand, under exchange conditions, the cAMP binding capacity of all subcellular fractions was similar in control and ACTH treated cells, except in the 175 000 x g pellet in which it was slightly decreased in ACTH-treated cells. These modifications were observed for supraphysiological (10(-8) M), as well as for physiological (10 (-11) M), concentrations of ACTH. These observations suggest that, after ACTH stimulation, a liberation of free catalytic sub-unit occurs from particulate into the soluble cell compartment, which shows an activation of particulate cAMP-dependent protein kinase activity. PMID- 6288493 TI - The mode of action of LHRH agonists on the rat Leydig cell. AB - The relationship between LHRH agonist-receptor interaction and subsequent stimulation of steroidogenesis has been investigated using dispersed adult rat Leydig cells. Binding of 125I-labelled LHRH agonist to these cells was rapid and was readily and completely reversible, which contrasted with the binding of 125I labelled hCG which was never completely reversible and which became progressively less reversible with increase in time. Following 125I-LHRH agonist-receptor interaction for 5 min at 21 degrees C, over 80% of the bound hormone dissociated at a fast rate (t 1/2 1.5 min) and the remainder at a slower rate (t 1/2 20 min). Following LHRH agonist-receptor interaction for a longer period (45 min) at 21 degrees C, only the slower of these 2 components of dissociation was evident, although binding was still completely reversible. Evidence was also obtained that following preincubation of Leydig cells for 2 hr at 34 degrees C with a near saturating concentration (2000 pg/ml) of unlabelled LHRH agonist, all or most of the bound hormone dissociated during subsequent incubation at 21 degrees C. Incubation of Leydig cells at 34 degrees C in the continuous presence of either a low dose (1 pM) of hCG or a high dose (2000 pg/ml) of LHRH agonist resulted in similar percentage stimulation of testosterone secretion with a similar time lag (2 h) before this increase was evident. Preincubation with this dose of hCG for as little as 15 or 30 min still led to increased testosterone secretion following removal of the free hormone. In contrast, preincubation with LHRH agonist for up to 2 h was without subsequent stimulatory effect, presumably due to dissociation of the bound hormone from its receptors. Therefore, for LHRH agonist to stimulate Leydig cell steroidogenesis in vitro, it must be present continuously in the incubation medium, and the implications of this with respect to the secretion and mode of action of 'testicular LHRH' in vivo are discussed. PMID- 6288494 TI - GnRH receptors in cultured rat granulosa cells: mediation of the inhibitory and stimulatory actions of GnRH. AB - The regulatory actions of FSH and the GnRH agonist [D-Ala6]des-Gly10-GnRH N ethylamide (GnRHa) upon ovarian GnRH receptors were studied in granulosa cells obtained from ovaries of hypophysectomized diethylstilbestrol-treated rats. When granulosa cells were cultured for 48 h in the presence of FSH (5-250 ng/ml) the binding of 125I-GnRHa to granulosa cell receptors was increased in a dose dependent manner, to a maximum of 3-4 fold above the control value. Addition of FSH (100 ng) also caused a dose-dependent increase of more than 100-fold in the accumulation of cAMP and progesterone in the culture medium. In freshly prepared cells, Scatchard analysis of GnRHa binding data revealed an equilibrium constant (Ka) of 1.1 x 10(10) M-1 and GnRH receptor concentration fo 401 fmoles/mg protein. Granulosa-cell GnRH receptors decreased during culture without FSH, but were maintained in the presence of 100 ng FSH at 580 femoles/mg protein, with Ka of 0.8 x 10(10) M-1. This action of FSH on GnRH receptors was significantly reduced by 10(-8) M GnRHa. Also, GnRHa concentrations of 10(-10) and 10(-8) M caused inhibition of FSH-induced cAMP and progesterone accumulation. In cells cultured with GnRHa alone, there was a slight enhancement of GnRH receptors by GnRHa concentrations up to 10(-8) M, and a decrease below control levels with higher amounts. Also, GnRHa concentrations from 10(-8) to 10(-5) M caused a 3-4 fold increase in cAMP accumulation in the absence of FSH. These results demonstrate that FSH maintains GnRH receptors in cultured granulosa cells, and that GnRHa attenuates this effect, as well as the other actions of FSH on granulosa cell maturation. It is also evident that GnRHa itself can slightly stimulate cAMP production and partially maintain GnRH receptors, but at high concentrations causes loss of the homologous receptor sites. These findings also emphasize the ability of GnRH agonists to exert both positive and negative direct effects on rat granulosa cell function. PMID- 6288496 TI - Spontaneous release of nucleosome cores from embryoblast chromatin. PMID- 6288495 TI - Molt cycle-associated changes in calcium-dependent proteinase activity that degrades actin and myosin in crustacean muscle. PMID- 6288497 TI - Induction of squamous cell carcinoma with 3,4-benzo[a]pyrene in the human bronchus transplanted into nude mice. AB - By the intraluminal injection of 3,4-benzo[a]pyrene into human bronchial grafts in nude mice, squamous cell carcinoma was induced in one bronchial graft out of 17 benzo[a]pyrene-treated grafts. As benzo[a]pyrene is one of the most common carcinogens in the human environment, this experimental system should serve as an excellent model system for studying the process of carcinogenesis in the human respiratory tract. PMID- 6288498 TI - Matrical inclusions of peroxisomes induced by clofibrate in preneoplastic hepatocytes of rats fed 3'-methyl-4-dimethyl-aminoazobenzene. AB - The response of peroxisomes to clofibrate in preneoplastic hepatocytes of rats fed 3'-methyl-4-dimethylaminoazobenzene was examined histochemically (using the 3,3'-diaminobenzidine reaction) and electron microscopically. Morphometrical examinations revealed that the number of peroxisomes correlated well with the intensity of the histochemical reaction. Preneoplastic hepatocytes which responded to clofibrate with a marked proliferation of peroxisomes showed no matrical inclusions or merely matrical plates in very low incidence in the proliferated peroxisomes. The cells that exhibited a moderate proliferation of peroxisomes showed formation of peroxisomal matrical inclusions in high incidence. The inclusions were matrical plates and tubules showing the same fine structures as those found in 2-acetylaminofluorene carcinogenesis. These peroxisomal inclusions were absent in the cells which responded to clofibrate with no or only a slight increase in the number of peroxisomes. It is suggested that these varieties of peroxisomal changes might reflect the progress of preneoplastic phases of the cells. PMID- 6288499 TI - Serological analysis of cell surface antigens on methylcholanthrene-induced mouse sarcomas with antisera produced against the tumors. AB - Out of 9 cell lines from methylcholanthrene-induced fibrosarcomas in BALB/c, 4 lines without expression of murine leukemia virus (MuLV) were selected for immunization of three different F1 hybrids with mitomycin C-treated in vivo sarcoma cells. Antibody activity was tested by immune adherence (IA) and protein A (PA) assays after the 8th immunization. Two sarcomas, CMC-11 and CMC-12, produced antibodies detectable by IA in (BALB/c x C3H/He)F1, but no antibodies were detected by PA. (BALB/c x C3H/He)F1 anti-CMC-11 was tested against CMC-11 and three other MuLV- sarcomas as well as against the skin fibroblast (C-11) from a BALB/c mouse, in which the primary CMC-11 sarcoma was induced. CMC-11 line reacted up to 1/80, but other lines were weakly positive or negative. Absorption analysis showed that the antigen defined by anti-CMC-11 was present on CMC-11 as well as on many other sarcoma cell lines, but not on BALB/c thymocytes and spleen cells, nor on leukemia and myeloma cells. Among 5 fibroblast cultures tested, C 14 showed complete absorption. It seems significant that only C-14 produced tumor nodules in BALB/c mice, in contrast to 4 other fibroblast cultures. This suggested that this antibody was directed against antigens present on transformed fibroblasts and sarcoma cells. The specificity of (BALB/c x C3H/He)F1 anti-CMC-12 serum indicated that this antiserum also seemed to detect transformation-related antigens, although it showed a broader specificity with regard to the tissue representation detected. PMID- 6288500 TI - Induction of hyperplastic liver nodules in hepatectomized rats treated with 3' methyl-4-dimethylaminoazobenzene, benzo[a]pyrene or phenobarbital before or after exposure to N-2-fluorenylacetamide. AB - The effects of pre- and post-administration of 3'-methyl-4 dimethylaminoazobenzene (3'-Me-DAB), benzo[a]pyrene (B[a]P), and phenobarbital (PB) on the induction of hyperplastic liver nodules by N-2-fluorenylacetamide (2 FAA) were studied in F344 rats. Rats were given 3'-Me-DAB (0.06% in the diet), B[a]P (80 mg/kg b.wt. x 7, intragastrically) or PB (0.05% in the diet) for 6 weeks before (experimental series I) or after (experimental series II) diet containing 0.02% 2-FAA for 2 weeks, and killed in week 10 of the experiment. The number and total area of hyperplastic nodules per unit area of liver were greatest in rats treated with 3'-Me-DAB in both experimental series. B[a]P significantly increased the induction of hyperplastic nodules in experimental series I and PB significantly increased it in experimental series II. Partial hepatectomy performed during administration of test chemicals enhanced the induction of hyperplastic nodules, especially in experimental series I, and an oral dose of carbon tetrachloride during 2-FAA feeding enhanced it in both experimental series. In the present system, initiating activity of both the hepato- and the non-hepatocarcinogen was detected, and promoting activity of both the hepatocarcinogen and the hepatopromoter was demonstrated. PMID- 6288501 TI - Therapeutic effect of an aromatic retinoic acid analog on rats with bladder carcinoma upon administration alone or in combination with mitomycin C. AB - An aromatic retinoic acid analog (Ro 10-9359) was given orally in combination with intraperitoneal administration of mitomycin C (MMC), in an attempt to reduce the toxicity and enhance the therapeutic effect. Male ACI/N rats were inoculated subcutaneously with BC50-TC cells, an established bladder carcinoma cell line of ACI/N rats. The chemotherapy was initiated at 16 days after the inoculation and continued for 4 weeks thereafter. Tumor growth was significantly inhibited in the rats given 100 mg/kg/week Ro 10-9359 or 0.3 mg/kg/twice a week MMC. Neither additive nor synergistic effect was apparent when these two drugs were given simultaneously. No effect was seen when 0.1 mg/kg/twice a week MMC was given alone or in combination with Ro 10-9359. MMC may suppress Ro 10-9359 activity directly or indirectly by affecting the responding cells. Therefore care should be taken when prescribing retinoids with MMC or possibly with other cytotoxic agents. PMID- 6288502 TI - Activities of various enzymes of pyrimidine nucleotide and DNA syntheses in normal and neoplastic human tissues. AB - The activities of the key enzymes of pyrimidine nucleotide and DNA syntheses in 43 human tumors and 28 normal human tissues were investigated. The activities of cytidine triphosphate synthetase, deoxycytidine monophosphate deaminase, uridine kinase, thymidine kinase, thymidine monophosphate kinase and DNA polymerase were markedly increased in tumor tissues, compared with those in the corresponding normal tissues, while the activities of deoxycytidine kinase, cytidine deaminase and deoxycytidine deaminase were only slightly increased. The use of thymidine and deoxyuridine as substrates of human pyrimidine nucleoside phosphorylase gave 1 to 2 orders of magnitude higher activity than that of uridine. PMID- 6288504 TI - [Hepatocellular carcinoma and primary biliary cirrhosis: a new case]. PMID- 6288503 TI - Asymptomatic type C virus carriers in the family of an adult T-cell leukemia patient. PMID- 6288506 TI - Colonic dysfunction in multiple sclerosis. AB - Multiple sclerosis is a central nervous system disease frequently accompanied by urinary symptoms and severe constipation. In order to investigate the pathophysiology of these symptoms, we studied colonic motor and myoelectrical activity, as well as colonic volume-pressure relationships (colonometrograms) and have correlated these data with cystometry and electrophysiologic studies of the central and peripheral somatosensory nervous system. The study group consisted of 7 patients with advanced multiple sclerosis marked by symptoms and signs of somatic and visceral nervous system dysfunction including severe constipation. Ten normal volunteers served as control subjects. The multiple sclerosis group demonstrated electrophysiologic evidence of lesions in the somatosensory neuroaxis central to the lumbosacral spinal cord. Abnormal cystometrograms suggested visceral central nervous system dysfunction. Colonometrograms in the multiple sclerosis group demonstrated a more rapid pressure rise than in the control group (p less than 0.01). The multiple sclerosis group failed to demonstrate the postprandial increase in colonic motor and myoelectrical activity observed in the control group (p less than 0.01). Abnormal colonometrograms and absent postprandial colonic motor and myoelectric responses may be features of visceral neuropathy in patients with advanced multiple sclerosis and severe constipation. PMID- 6288505 TI - Diagnostic ability of hepatobiliary scintigraphy. AB - Morphological aspects of biliary images were observed by the hepatobiliary scintigraphy using 99mTc-PI (pyridoxylidene isoleucine). Our study was made mainly on the diagnostic ability of the scintigram in various hepatobiliary diseases. Diagnostic findings were observed in 53.8% of the cases with intrahepatic stones and all cases with cholangioma. In the stone of the common bile duct and extrahepatic biliary cancer, the diagnostic findings were observed in 42.3% and 37.5% respectively. Hepatobiliary scintigraphy was less reliable for the diagnosis of the lesions in the gallbladder. The findings by hepatobiliary scintigraphy were as same as those by X-ray basically. In the cases showing the diagnostic finding by hepatobiliary scintigraphy, it seems to be possible to distinguish the stone from the cancer. Contrary to X-ray picture, scintigram is inferior in the image. So, it is necessary to perform ERCP or PTC to investigate fully the hepatobiliary lesions. PMID- 6288508 TI - Prolonged survival with hepatocellular carcinoma. PMID- 6288507 TI - Clinical value of the determination of serum guanase activity. Studies on patients and experimental data from mongrel dogs and cultured rat hepatocytes. AB - Serum guanase activity was measured by a new method and compared with serum glutamic oxalacetic transaminase and glutamic pyruvic transaminase levels in 150 patients with various disorders, 21 dogs with experimental myocardial infarction, and 2 CCl4-treated dogs. Additionally, studies of the effect of CCl4 on enzyme release were undertaken using cultured rat hepatocytes. Glutamic oxalacetic transaminase, glutamic pyruvic transaminase, and guanase activities were found to be significantly elevated in patients with various liver disorders, those with acute myocardial infarction with prominent congestion of the liver, and also in CCl4-treated dogs. However, serum guanase activity was normal in patients with a variety of non-liver-related diseases including acute myocardial infarction, and in dogs with experimental myocardial infarction without liver damage, even when the serum glutamic oxalacetic transaminase and glutamic pyruvic transaminase activities were increased. The glutamic oxalacetic transaminase, glutamic pyruvic transaminase, and guanase activities in the culture medium of rat hepatocytes indicated in the presence of 0.5 mM CCl4 were elevated. These findings indicate that serum guanase activity is a more specific indicator of liver damage than serum glutamic oxalacetic transaminase and glutamic pyruvic transaminase. PMID- 6288509 TI - Isosorbide dinitrate and nifedipine treatment of achalasia: a clinical, manometric and radionuclide evaluation. AB - The effects of sublingual isosorbide dinitrate (5 mg) and nifedipine (20 mg) were compared in 15 patients with achalasia. The parameters examined included the manometric measurement of the lower esophageal sphincter pressure, the radionuclide assessment of esophageal emptying and the clinical response. The mean basal lower esophageal sphincter pressure fell significantly after both drugs (p less than 0.01), with a maximum fall of 63.5% 10 min after receiving isosorbide dinitrate, but by only 46.7% 30 min after nifedipine. The esophageal radionuclide test meal retention was significantly less (p less than 0.01) only after receiving isosorbide dinitrate. The drug improved initial esophageal emptying by its effect on the lower esophageal sphincter and by relieving the test meal hold-up noted to occur at the junction of the upper and midesophagus. Eight patients cleared their test meal within 10 min after isosorbide dinitrate administration while only two did so after nifedipine. Subjectively, 13 patients had their dysphagia relieved by isosorbide dinitrate and 8 by nifedipine. However, this relief was not confirmed in 4 patients by the radionuclide study and they, as well as the other 3 patients who did not respond to therapy, were referred to pneumatic dilatation. Side effects were more prominent after nitrates. Three of the patients are currently receiving nifedipine and 5 patients received isosorbide dinitrate therapy for 8-14 mo. The radionuclide test meal is currently the best way of objectively evaluating drug therapy in patients with achalasia. Isosorbide dinitrate is more effective than nifedipine in relieving their symptoms. PMID- 6288510 TI - Nodular growth on the tongue. PMID- 6288511 TI - The mode of action of the crustacean neurosecretory hyperglycemic hormone (CHH). II. Involvement of glycogen synthase. PMID- 6288512 TI - Stimulation of interrenal secretion in amphibia. I. Direct effects of electrolyte concentration on steroid release. PMID- 6288513 TI - A radioimmunoassay for 17 alpha 20 beta-dihydroxy-4-pregnen-3-one: its use in measuring changes in serum levels at ovulation in atlantic salmon (Salmo salar), coho salmon (Oncorhynchus kisutch), and rainbow trout (Salmo gairdneri). PMID- 6288514 TI - Estimating genetic variability with restriction endonucleases. AB - The estimation of the amount of sequence variation in samples of homologous DNA segments is considered. The data are assumed to have been obtained by restriction endonuclease digestion of the segments, from which the numbers and frequencies of the cleavage sites in the sample are determined. An estimator, p, of the proportion of sites that are polymorphic in the sample is derived without assuming any particular population genetic model for the evolution of the population. The estimator is very close to the EWENS, SPIELMAN and HARRIS (1981) estimator that was derived with the symmetric WRIGHT-FISHER neutral mode. ENGELS (1981) has also recently proposed an estimator of the same quantity, and he arrived at his estimator without assuming a particular population genetic model. The sampling variance of p and ENGELS' estimator are derived. It is found that the sampling variance of p is lower than the sampling variance of ENGELS' estimator. Also, the sampling variance of theta, an estimate of theta (=4Nu) is obtained for the symmetric WRIGHT-FISHER neutral model with free recombination and with no recombination. PMID- 6288515 TI - [Fibrogenic activity of serpentinite dust containing chrysotile asbestos in an experimental study]. PMID- 6288516 TI - [Spread of central cancer of the lung along the bronchial wall in a proximal direction]. PMID- 6288517 TI - Effects of selective beta-adrenergic agonists on spontaneous contractions, cAMP levels and phosphodiesterase activity in myometrial strips from pregnant women treated with terbutaline. AB - The beta-adrenergic receptor function in terms of cAMP production and phosphodiesterase activity has been studied in myometrial strips taken from pregnant women at cesarean sections. The material consists of 10 patients treated with terbutaline for threatening premature delivery and an untreated control group of 10 patients. The basal levels of cAmP were lower and the cAMP production after receptor stimulation in vitro less in myometrial strips taken from terbutaline-treated women compared to the untreated control group. The phosphodiesterase activity was higher in myometrial strips taken from terbutaline treated women, indicating a faster rate of degradation of cAMP in these women. PMID- 6288519 TI - Demonstration of human papilloma virus antigen in the condylomatous lesions of the uterine cervix by immunoperoxidase technique. AB - Routine histological sections of characteristic condylomas (showing dysplasia of different degrees) collected from the uterine cervix of 10 women were subjected to staining with indirect immunoperoxidase technique using antihuman papillomavirus (anti-HPV) serum to demonstrate the HPV antigens in these lesions. The anti-HPV immune serum was raised in guinea pigs immunized with highly purified viruses from a pool of skin wart lesions collected from various anatomical sites. 8 out of the 10 condylomas studied showed a definitely positive staining (brown precipitate) with the HPV anti-serum, indicating the presence of HPV antigens within the nuclei of a few cells (mostly koilocytes and dyskeratotic cells) near to the surface of the lesions. The present technique provides further confirmation to the HPV etiology of cervical condylomas, and seems to offer a definite advantage as a diagnostic tool of these lesions and other squamous cell lesions as well. The application of this technique on malignant tumors will most probably cast light on the possible role of HPV in the etiology of human squamous cell carcinomas in the near future. PMID- 6288518 TI - Impaired pregnenolone secretion after combined cyproterone acetate and ethynyl estradiol therapy in hirsute patients. AB - 6 women affected by hirsutism, either of idiopathic origin or due to polycystic ovary syndrome, have been treated with cyproterone acetate and ethynyl estradiol in combined therapy using, respectively, 100 mg and 50 micrograms/day, from the 5th to the 25th day of the cycle. The adrenal function was assessed before treatment and at the end of the 4th month of therapy, evaluating the peripheral plasma concentrations of pregnenolone (delta 5P), progesterone, 17-OH progesterone, dehydro-epiandrosterone sulfate, androstenedione, testosterone, and cortisol in basal conditions and after dexamethasone suppression and an adrenocorticotropic hormone (ACTH) stimulation test. A group of healthy, untreated females were examined in the early follicular phase, as controls, Before therapy, the hirsute patient showed testosterone and androstenedione plasma levels, which were significantly higher than in the controls, and a significant reduction in pregnenolone response to ACTH. After 4 months of therapy with cyproterone acetate plus ethynyl estradiol, a significant decrease was found in testosterone and androstenedione plasma levels, and pregnenolone basal plasma levels, dexamethasone suppressibility, and response to ACTH were also markedly reduced, showing a significant difference versus the same patients before therapy and versus the control group. The existence of an impairment in adrenal function after cyproterone acetate plus ethynyl estradiol therapy at the given dose seems to be evident only in the case of directly ACTH-dependent adrenal enzymatic activities responsible for cholesterol cleavage to pregnenolone. PMID- 6288520 TI - Effect of the immunological antigenicity of the allogeneic tendons on tendon grafting. PMID- 6288521 TI - Synovial sarcoma of the finger. PMID- 6288522 TI - Freeze-fracture study of the red cells and red cell precursors of a patient with congenital inclusion body anaemia. AB - Comparison of thin sections and freeze fracture replicas of red cells and red cell precursors from the peripheral blood of a patient suffering from congenital inclusion body anaemia provided new morphological information on Heinz body substructure, Heinz body-membrane attachment, intracellular membrane formation, as well as on membrane fusion and orientation of intracellular membranes. It is suggested that the fusion of autophagic vacuoles with the plasma membrane may contribute to the increased haemolysis of red cells in inclusion body anaemia. PMID- 6288523 TI - [Popliteal neuritis complicating temporal arteritis]. PMID- 6288524 TI - [The role of prostaglandins and the processes of reproduction]. PMID- 6288525 TI - [Synovial sarcoma mistaken for metastasis of follicular thyroid carcinoma: a cautionary tale]. PMID- 6288526 TI - [Early clinical presentation of synovial sarcoma]. PMID- 6288527 TI - [Hypercalcemia in malignant diseases]. PMID- 6288528 TI - [Should we be wary of a high-fiber diet?]. PMID- 6288529 TI - [Rotavirus infection in infants]. PMID- 6288530 TI - [Toward a rotavirus vaccine]. PMID- 6288531 TI - [Malignant fibrous histiocytoma of bone (MFH)--an new tumour? ]. AB - Malignant fibrous histiocytoma of bone (MFH) is a histologically defined bone tumour, which can be distinguished from osteo- and fibrosarcomas. The present paper deals with the question whether MFH can be distinguished from these tumours on clinical grounds. One hundred and fifty-seven previously published, and four additional cases have been analysed with respect to their localisation and their biological, clinical and radiological behaviour and their prognosis. It was found that MFH can be distinguished from osteosarcomas on clinical grounds and that this is significant in relation to treatment; its distinction from fibro-sarcomas is clinically not justified. PMID- 6288532 TI - [Skeletal scintigraphy for the observation of course and treatment of carcinoma of the prostate ]. AB - Serial skeletal scintigraphy was carried out on 259 patients with carcinoma of the prostate; 5-10% developed bone metastases each year. Isotope uptake was measured in 79 patients receiving treatment; in 60.6% (48 patients) there was a reduction or disappearance of the areas of uptake, in 16.6% (13 patients) they were stationary and 22.8% (18 patients) the metastases progressed. Skeletal scintigraphy is therefore useful during the course of the disease in showing its extent and the effect of treatment. PMID- 6288535 TI - [Status asthmaticus]. PMID- 6288533 TI - [Capillary embolization. Part II. Occlusion of the entire arterial system of experimentally induced renal tumors ]. AB - 221 renal rat tumors (induced by Dimethylnitrosamine) are embolized by gelfoam powder, IBC/Lipiodol and Ethibloc/Glucose. The effect of those embolization media is compared to ligation and glucose perfusion. Recurrencies and rate of surviving tumor tissue are dependent on the stage of the neoplasm, sufficient capillary propagation of the medium, and exact volume-dependent embolization. It could be proven that large areas of renal rat tumors are equally supplied by the main artery and parasitary collaterals. Only homogeneous distribution and complete downstream propagation of the embolization medium from the main artery to the capillaries will achieve breakdown of this parasitary supply. Glucose governed Ethibloc embolization achieves in 80% of tumors with a 6 fold renal volume and in 50% of larger tumors complete necrosis. It has the lowest complication rate of all tested materials. PMID- 6288534 TI - [Extracellular electrolytes and arterial hypertension. New pathogenetic and therapeutic aspects]. PMID- 6288536 TI - [Investigations of the pathogenesis of lung cancer observed among chromate factory workers]. AB - In order to clarify the relationship between the chromate compound and occurrence of lung cancer, the author studied the characteristics of patients with lung cancer among workers of chromate factory and measured the chromium contents of each tissues obtained from 14 patients at surgery and autopsy. The incidence of the chromate lung cancer was 413 per 100,000 population, which was 16 times that of the general population. All were male. The age ranged from 26 to 74 year old (average 53). The histological type was mostly squamous and small cell carcinoma. Location of carcinoma occurrence was mainly limited to the large bronchi. The average total labor period of patients with lung cancer was 258 months and the latent period was 305 months. The working history of the patients in hexavalent chromium producing process was longer than that of the control group. Patients with small cell carcinoma was mainly engaged in the monochromate producing process. The labor and latent period of patients with squamous cell carcinoma was longer than those of small cell carcinoma. Measurement of chromium contents in the respiratory system of chromate workers revealed much higher chromium content than in control group. The chromium content of tissues in the non-respiratory system was a little higher than that of the control group. High chromium content itself did not have any relation with the occurrence of lung cancer because the primary location of chromate lung cancer was limited to the large bronchi, not to the peripheral lungs which contained the highest chromium content. The longer was the exposure period, the higher was chromium content in the lung. Chromium content in the upper lobe was higher than that in the lower lobe. From these studies, the author concluded that hexavalent-chromate compound might be the compound responsible for lung cancer occurrence. PMID- 6288537 TI - Glucocorticoid-induced changes in insulin secretion related to the metabolism and ultrastructure of pancreatic islets. AB - The effect of adrenalectomy and dexamethasone-treatment on insulin secretion was studied and related to the changes observed in the glucose oxidation, calcium uptake, cAMP and insulin content, as well as the ultrastructure of pancreatic rat islets. It was found that adrenalectomy was followed by a decreased glucose induced insulin secretion, glucose oxidation, calcium uptake, cAMP and insulin content without any remarkable change observed at the ultrastructural level. Conversely, adrenalectomized-rats supplemented with dexamethasone showed an increased glucose-induced insulin secretion, glucose oxidation, calcium uptake and cAMP content but a diminished islet insulin content. At the ultrastructural level, a clear picture of increased secretory activity was found, with diminished number of mature B granules and greater number of pale granules, while rough endoplasmic reticulum and Golgi complex frequently appeared hypertrophic. These changes were only observed in the B cells. On account of our results, we might suggest that insulin secretion is partially controlled by glucocorticoid circulating levels throughout their effect on pancreatic islet metabolism. PMID- 6288538 TI - Insulin and beta receptor modulation of K homeostasis in nephrectomized dogs with hyperkalemia. AB - In nephrectomized dogs infused with 2 mEq KCl/kg/hr a homeostatic mechanism retards the development of hyperkalemia by transferring about 70% of the K load to intracellular fluid. beta Adrenergic receptor activity is importantly involved in the transfer process; halting it with propranolol reduces the proportion transferred to less than 35%. The addition of pancreatectomy increases the involvement of beta receptor activity; propranolol treatment now reduces the proportion transferred to less than 20%. Insulin treatment, on the other hand, not only improves transfer of a K load, it also alters the response to propranolol. Nephrectomized dogs treated with 2 U insulin/kg/hr deposit some 80% of the infused K in intracellular fluid. After beta receptor blockade, nearly 90% is transferred. The results suggest that in the K homeostatic mechanism of nephrectomized dogs, insulin and beta receptors may be reciprocally related. K transfer mediated by beta receptors improves after pancreatectomy, and insulin mediated K transfer improves after beta receptors are inactivated. PMID- 6288539 TI - Streptozotocin treatment of a juvenile onset type diabetic patient with Verner Morrison syndrome and multi hormonal probable malignant islet cell carcinoma with liver metastases. PMID- 6288540 TI - Plasma 25-hydroxyvitamin D3 response to a pharmacological dose of vitamin D3 or 25-hydroxyvitamin D3 during chronic ethanol administration in the rat. AB - Plasma 25-(OH)D3 concentrations following an intra-portal injection of 100 micrograms Kg-1 of D3 or 100 micrograms Kg-1 of 25-(OH)D3 was studied in D depleted rats fed ethanol diet and pair-fed controls. When challenged with D3, the rats under ethanol feeding were unable to increase their plasma 25(OH)D3 concentrations above those observed in controls. Plasma 25(OH)D3 concentrations following 25(OH)D3 administration were however lowered by the ethanol treatment 3 and 96 hr after 25(OH)D3 administration (p less than 0.05). These results suggest that animals chronically exposed to ethanol have an unaltered plasma 25(OH)D3 response following a pharmacological dose of D3 while the drug treatment contributes to an accelerated plasma 25(OH)D3 disappearance following 25(OH)D3. The former observations also suggest that D3 does not seem to be a high affinity substrate for the ethanol-induced cytochrome P-450. PMID- 6288541 TI - Radioimmunoassay for 4-hydroxyestrone 4-methyl ether in human urine. AB - 4-Hydroxyestrone 4-methyl ether (4-OHE1 4-Me) was converted to its 17-(O carboxymethyl)oxime and then coupled to bovine serum albumin. The injection of this steroid-protein conjugate into rabbits induced the formation of antibodies with high specificity and affinity for 4-OHE1 4-Me. With this antiserum a radioimmunoassay was developed which allowed the measurement of 4-OHE1 4-Me with a lower limit of detection of 6 pg/tube. Using a simple and practicable method for the hydrolysis and purification of urine, the excretion rates of 4-OHE1 4-Me were reliably measured in healthy human subjects: male children 0.1 microgram/24 h, female children 0.2 micrograms/24 h, men (20-45 years) 0.7 micrograms/24 h, men (greater than 50 years) 0.5 micrograms/24 h, women, follic. 0.5 micrograms/24 h, periov. 0.6 micrograms/24 h, luteal 0.6 micrograms/24 h, women pregn., first trim. 2.3 micrograms/24 h, sec. trim. 2.9 micrograms/24 h, third trim. 5 micrograms/24 h, women postmenop. 0.5 micrograms/24 h. These urinary excretion rates of 4-OHE1 4-Me are significantly lower than those of 4-hydroxyestrone. Comparing the ratios 4-OHE1 4-Me/4-hydroxyestrone with those of 2-hydroxyestrone 2-methyl ether/2-hydroxyestrone, it becomes obvious that endogenous 4 hydroxyestrogens are methylated in vivo to a much lesser extent than the isomeric 2-hydroxyestrogens, a finding which could partly explain why 4-hydroxyestrogens have higher biologic potencies than their 2-hydroxylated isomers PMID- 6288543 TI - Adrenalectomy reduces exploratory activity in the rat: a specific role of corticosterone. PMID- 6288542 TI - Geographic distribution of HBsAg carriers in China. AB - We tested by radioimmunoassay the sera of 22,707 male Chinese government employees in Taiwan for HBsAg and found 15.2% to be positive. Almost all were confirmed as carriers by follow-up testing 1 year later. This paper presents HBsAg positivity rates of these men according to the province of China from which they originated, and compares the rates with published provincial mortality rates for primary hepatocellular carcinoma. Moderate variation in carrier rates between provinces was observed with high prevalence in men from provinces south of the Yangtze River. There was positive correlation between HBsAg prevalence and primary hepatocellular carcinoma incidence giving further support to the possibility of a causal relationship. PMID- 6288544 TI - The pathogenesis of infectious mononucleosis. PMID- 6288545 TI - Selective effects of ionizing radiations on immunoregulatory cells. PMID- 6288546 TI - Cyclosporin A--usefulness, risks and mechanism of action. PMID- 6288547 TI - Isolation of xanthophores from the goldfish (Carassius auratus L.). AB - A method is described for the isolation of milligram quantities of viable, hormone-responsive xanthophores from goldfish scales. The preparations are typically 70 to 90% pure and are useful for biochemical analyses or for establishing primary cultures. PMID- 6288548 TI - Rat serum stimulates serotonin N-acetyltransferase activity in pineal monolayer cultures. AB - The effects of rat serum on serotonin N-acetyltransferase (NAT) activity and indole synthesis in monolayer cultures of neonatal rat pineal glands was examined. The addition of 5% rat serum to these cultures resulted in stimulation of NAT activity equal to that obtained with optimal concentrations of the adrenergic agonist norepinephrine (NE). Rat serum also increased the synthesis of both N-acetylserotonin and melatonin from tryptophan. Stimulation of NAT activity by rat serum was partially blocked by metoprolol and propranolol, but not by phentolamine or butoxamine. The serum factor responsible for the stimulation was stable to both freezing and boiling. No significant amounts of epinephrine, norepinephrine, or dopamine were detected in the serum. PMID- 6288550 TI - Cholera toxin stimulation of human mammary epithelial cells in culture. PMID- 6288549 TI - Angiotensin I-converting enzyme localization on cultured fibroblasts by immunofluorescence. AB - Angiotensin I-converting enzyme is responsible for the activation of angiotensin I and the inactivation of bradykinin. It has been localized by immunofluorescence on the endothelium of a variety of tissues and has been considered to be a specific marker for endothelial cells in culture. The present paper demonstrates, by immunofluorescence, the presence of angiotensin I-converting enzyme in monolayer cultures of fibroblasts derived from adult rat lung, bovine calf pulmonary artery, and human foreskin (CF-3 cells). Fluorescent localization of angiotensin I-converting enzyme was observed over the cytoplasm of adult rat lung and bovine calf pulmonary artery fibroblasts and as distinct areas overlying the nuclei of human foreskin fibroblasts. Determination of angiotensin I-converting enzyme activity by fluorimetric assay in parallel studies confirmed the presence of angiotensin I-converting enzyme activity in cultured fibroblasts. Immunofluorescent studies with antibody to Factor VIII demonstrated the presence of Factor VIII on cultured endothelial cells but not on fibroblasts. These results indicate that angiotensin I-converting enzyme is not confined to endothelial cells, and thus may not serve as a specific marker for endothelial cells in culture. Factor VIII may be a more specific marker for these cells. PMID- 6288551 TI - Continuous production of anti-host IgG antibodies contained in circulating IgG anti-IgG complexes. AB - Weekly injection with antibody-coated pertussis led to the chronic production of serum complexes in mice. Mice bearing one allotype. Iga, received spleen cells from a congenic strain bearing another allotype, Igb, and weekly injections with Iga-coated pertussis. The serum complexes from these mice and from those receiving challenges alone were separated by ultracentrifugation at neutral and acid pH on sucrose density gradients and their fractions tested for their anti Iga and Iga content. This revealed the presence of anti-host (Iga) allotype antibodies in fractions from mice that had received Igb cells but not from mice given Iga-coated pertussis alone. Iga allotype was detected in fractions from both groups. It is considered that anti-host allotype antibodies are continuously produced but that they cannot be detected in unfractionated serum because the antibody forms complexes with Iga. These findings are discussed in relation to rheumatoid disease in humans. PMID- 6288552 TI - Thermostability & turnover of phosphatases in the obligate thermophile Thermoactinomyces vulgaris. PMID- 6288553 TI - Genetic influences on the control of nociceptive responses & precipitated abstinence in mice. PMID- 6288554 TI - c-AMP-phosphodiesterase activity in fat body & testes of male rats & effect of aspirin on the same. PMID- 6288555 TI - Localization of Chandipura virus multiplication sites in tracheal cultures. PMID- 6288556 TI - Neurotoxic effect of patulin. PMID- 6288557 TI - Increased nociceptive reactivity in mice after the administration of newer opioid antagonists--further evidence for a role of endogenous opioids. PMID- 6288559 TI - Interference of Salivary immunoglobulin A antibodies and other salivary fractions with adherence of Streptococcus mutans to hydroxyapatite. AB - The adherence of Streptococcus mutans to hydroxyapatite was studied in the presence of salivary fractions with varying activity of naturally occurring immunoglobulin A (IgA) antibodies. Human parotid saliva from different donors was fractionated by chromatography and compared. Salivary IgA antibodies had no decisive effect on the adherence of the S. mutans strain used. High-molecular weight salivary components from some subjects had an adherence-promoting effect, whereas fractions collected after the void volume of a Sepharose 2B column always inhibited adherence. The data indicate that the influence of unfractionated saliva on adherence is dependent on the net effect of adherence-promoting and adherence-inhibiting components. This principle has to be considered when the effect of human saliva on microbial adherence is studied. PMID- 6288560 TI - Experimental infection and immune response of guinea pigs with varicella-zoster virus. AB - An immune response (fluorescent antibody to membrane antigen) was detected in guinea pigs inoculated with varicella-zoster virus (VZV) adapted to guinea pig embryonic cells, including the Oka vaccine strain, even when inoculation was by an external route, i.e., nasal or corneal. Live or UV-inactivated virus having the same virus titer before irradiation was administered to guinea pigs by the corneal route, and antibody induction was detected only with live virus. The transmission of VZV from infected guinea pigs to noninfected ones was suggested by the appearance of antibody in the serum of the latter, who were kept in the same cage. The time course of the appearance of humoral and cellular immune responses in guinea pigs was examined by the fluorescent antibody to membrane antigen test and the skin reaction, with varicella antigen representing delayed type hypersensitivity. When VZV was injected subcutaneously, skin reaction appeared as early as 4 days after inoculation, which preceded the appearance of detectable antibody by 2 to 6 days. In in vitro studies, the Oka vaccine showed a higher adsorption rate and better growth in guinea pig embryonic cells than did other wild-type strains when assayed by the infectious center assay. These results suggest that a system of VZV adapted to guinea pig cells and guinea pigs provides a good animal experimental model for immunological study of VZV infection. PMID- 6288562 TI - Defective interfering particles of respiratory syncytial virus. AB - A multiplicity-dependent interference was observed in respiratory syncytial virus preparations (Randall strain) grown in HEp-2 cells, and the factor mediating this interference was characterized. Cloned virus did not demonstrate this multiplicity-dependent interference, but its replication was shown to be inhibited by the interfering factor by assays of reduction of infectious yield assay, the interferon factor was found to be particulate, to be inactivated by UV irradiation, and not to interfere with the replication of a heterologous virus, vesicular stomatitis virus. These characteristics are compatible with the physical properties and biological behavior of defective interfering particles. Defective interfering particles were generated by four undiluted passages of cloned virus but were not apparent after eight passages at a multiplicity of infection of 0.1. PMID- 6288561 TI - Toxic effect of human polymorphonuclear leukocytes on Chlamydia trachomatis. AB - The effect of human polymorphonuclear leukocytes (PMNs) on Chlamydia trachomatis was studied. Both trachoma (B/TW-5/OT) and lymphogranuloma venereum (L2/434/Bu) biotypes were rapidly inactivated by exposure to human PMNs. A decrease of 3 to 3.5 logs in viable count was observed after 60 min of incubation at a chlamydia to-PMN ratio of 1:10. Both chlamydial biotypes were also rapidly inactivated by the cell-free myeloperoxidase-H2O2-halide system. A decrease in infectivity titer of 4 to 5 logs for TW-5 and complete inactivation of 434 were seen after 30 min of incubation. The microbicidal effect was prevented by the deletion of each component of the system or by the addition of the peroxidase inhibitors cyanide or azide. PMNs from myeloperoxidase-deficient patients inactivated chlamydiae normally, whereas PMNs from patients with chronic granulomatous disease, although strongly chlamydicidal, were less effective than normal PMNs in the activation of TW-5 (2-log drop in viable organisms versus a 3 to 3.5-log drop). The chlamydicidal activity of PMNs from patients with chronic granulomatous disease and normal PMNs were comparable against the 434 biotype. These studies suggest that the myeloperoxidase system, or indeed oxygen-dependent antimicrobial systems, are not essential for the chlamydicidal activity of PMNs. PMID- 6288563 TI - Immunoglobulin M responses to the Norwalk virus of gastroenteritis. AB - Eighty-seven serum specimens from 20 human subjects experimentally inoculated one or more times with Norwalk virus were quantitatively examined for virus-specific immunoglobulin M (IgM). A sensitive and specific radioimmunoassay for anti Norwalk virus blocking activity was applied to whole serum and to separate IgM and IgG fractions obtained by sucrose density gradient ultracentrifugation. The peak IgM response occurred at about 2 weeks after illness, but IgM was detectable at lower titers for up to 21 weeks after infection. The IgM response was seen in volunteers who became ill, whether or not prechallenge total serum antibody was present. On long-term (27 to 42 months) rechallenge, volunteers who were previously ill and had produced IgM antibody again developed illness, and a secondary IgM response greater than the first was detected. Inoculated volunteers who did not develop illness, as well as previously ill volunteers on short-term rechallenge (4 to 14 weeks), usually failed to generate an IgM response, whether or not an IgG response had occurred. In ill subjects, the rise in IgM and IgG occurred concomitantly. Virus-specific IgM is not necessarily indicative of primary infection with Norwalk agent inasmuch as reinfection produces an enhancement of the IgM response. Furthermore, Norwalk-specific IgM responses do not appear to be associated with subclinical illness. PMID- 6288565 TI - Isolation of a fibronectin-binding protein from Staphylococcus aureus. AB - Fibronectin ("cold-insoluble globulin") has been suggested to play a role in cell to-cell and cell-to-substratum adhesions. The 70-kilodalton terminal part of human fibronectin has recently been shown to bind to Staphylococcus aureus. In the present study, a fibronectin-binding protein was purified from sonicated S. aureus strain E2371 by affinity chromatography on fibronectin-Sepharose. The fibronectin-binding protein was isolated from an extract of sonicated S. aureus containing at least 57 different proteins as determined by crossed immunoelectrophoresis in antibodies to sonicated S. aureus. The fibronectin binding protein was released from fibronectin-Sepharose by carbamide (8 M). No impurities in the final preparation could be detected when tested in crossed immunoelectrophoresis. By polyacrylamide gel electrophoresis in both reduced and unreduced gels, the protein showed two bands with relative molecular masses of 197,000 and 60,000, respectively. A complex between the purified S. aureus protein and fibronectin could be demonstrated by crossed immunoelectrophoresis both in monospecific antibodies against fibronectin and in S. aureus polyspecific antibody. PMID- 6288564 TI - Quantitation of binding and subcellular distribution of Clostridium perfringens enterotoxin in rat liver cells. AB - Binding of enterotoxin from Clostridium perfringens type A was studied in suspensions of parenchymal and nonparenchymal cells from rat liver. In hepatocytes, 1.5 X 10(6) specific binding sites per cell with an association constant of 3.2 X 10(6) M-1 were found. About 1% of the added toxin was nonspecifically bound to the hepatocytes. At concentrations of toxin below 0.1 micrograms/ml, 80% of the toxin density of 7 X 10(6) cells per ml. Binding did not increase after the cells became permeable to the toxin. Subcellular fractionation in a sucrose gradient produced no evidence for binding to parts of the cell other than the plasma membrane. The degree of binding to nonparenchymal cells was less than 10% of the binding to hepatocytes. PMID- 6288566 TI - Binding characteristics of wild mouse type C virus to mouse spinal cord and spleen cells. AB - Binding characteristics of mouse spinal cord and spleen cells to naturally occurring, ecotropic (paralytogenic and lymphomagenic 1504M virus) and amphotropic (lymphomagenic 1504A virus) retroviruses of wild mice were investigated. 125I-labeled ecotropic (N-tropic) virus bound efficiently to both spinal cord and spleen cells from SWR/J mice (Fv-1n), but labeled amphotropic (N tropic) virus bound efficiently only to spleen cells. The extent of binding of 1504M virus to the spinal cord cells was related to the time of incubation and to the amount of labeled input virus. 1504M virus bound to both glial and neuronal subpopulations of the spinal cord with almost equal efficiency. Binding of 125I labeled 1504M virus to SWR/J mouse spinal cord cells was competitively inhibited by unlabeled homologous virus, whereas an excess of unlabeled 1504A virus inhibited only 10% of the ecotropic virus binding. Unlabeled 1504M virus almost completely inhibited the low-level binding of 125I-labeled 1504A virus to spinal cord cells. The different extent of binding of 1504M virus to spinal cord cells from different strains of mice (SWR/J, NIH Swiss, BALB/c-Jax, Lake Casitas wild, and CD-1) correlated with the susceptibility to paralysis in these strains of mice after inoculation with 1504M virus. Although the spinal cord cells of BALB/c mice contained a moderate amount of 1504M virus receptor sites, these mice were resistant to virus-induced paralysis because of their Fv-1b genotype. Our results indicate that the receptor sites for wild mouse ecotropic retrovirus are strain and organ specific and suggest that the presence of such receptors in central nervous system tissue may be a prerequisite for a successful virus infection and paralysis induction in Fv-1n mice. PMID- 6288567 TI - Passive immunity in calf diarrhea: vaccination with K99 antigen of enterotoxigenic Escherichia coli and rotavirus. AB - Twenty-four pregnant cows were vaccinated intramuscularly with K99 extract from enterotoxigenic Escherichia coli and inactivated rotavirus as follows: six cows were injected with 2 ml of oil-adjuvanted vaccine; six cows were injected with 0.5 ml of oil-adjuvanted vaccine; six cows were injected with 4 ml of aluminum hydroxide-adjuvanted vaccine twice with a four-week interval; and six cows were unvaccinated as controls. Calves born to these cows were challenged with enterotoxigenic E. coli at 6 to 18 h after birth. Serum and milk antibodies to K99 and rotavirus in cows vaccinated with either dose of oil vaccine were significantly increased until at least 28 days after calving. In cows vaccinated with alhydrogel vaccine, there was a significant K99 antibody increase in serum and in colostrum but not in milk and a significant rotavirus antibody increase only in colostrum. Five of six calves born to unvaccinated cows developed enterotoxic colibacillosis after challenge, and all excreted the challenge strain of enterotoxigenic E. coli. None of the 18 calves in the three vaccinated groups developed clinical colibacillosis, and fecal excretion of the challenge organism was reduced. A combined enterotoxigenic E. coli-rotavirus vaccine may prove useful in preventing some outbreaks of calf diarrhea. PMID- 6288568 TI - Antigenic groupings of 90 rhinovirus serotypes. AB - We have completed production in rabbits of potent antisera to the 90 classified rhinovirus serotypes by using methods previously described (M. K. Cooney and G. E. Kenny, Proc. Soc. Exp. Biol. Med. 133:645-650, 1970). Systematic testing by neutralization tests has revealed significant numbers of cross-relationships among rhinovirus types, some of which have already been reported. Herein, our observations are compared with cross-reactions reported in National Institutes of Health reference guinea pig antisera. Also, original rhinovirus isolates, representing serotypes known to be antigenically related to other rhinoviruses, were tested against rabbit antisera to the related serotypes. These tests revealed extensive antigenic variation among isolates identified as rhinovirus 12:78 or 36:58, which are reciprocally related pairs, 41, reciprocally related to 13, and 67, which is related to both 9 and 32. If the rhinovirus serotypes were grouped according to antigenic relationships, 50 types could be included in 16 groups. PMID- 6288570 TI - Role of antiviral antibodies in resistance against coxsackievirus B3 infection: interaction between preexisting antibodies and an interferon inducer. AB - An experimental model of coxsackievirus B3 infection in newborn mice was utilized to examine the protective role of antiviral antibodies and an interferon inducer, polyinosinic acid-polycytidylic acid [poly(I:C)]. Subcutaneous administration to the infected mice of specific antiviral antibodies resulted in significant protection against coxsackievirus B3 infection. Antibody-treated animals had shortened viremia, early clearance of virus from tissues, and a reduced mortality rate. Dose response to antibodies was clearly demonstrated. However, the time of antibody administration in relation to the infection cycle was important. The protection was observed if antibodies were given before infection (24 h) or shortly after (2 h) infection. Administration of antibodies 24 h after infection resulted in no protection. The interferon inducer poly(I:C) prolonged the survival time of the infected mice, but this protective effect was incomplete and could only be demonstrated in animals treated before infection (24 h) or shortly after (2 h) infection. Enhanced protection against lethal coxsackievirus B3 infection was achieved in animals treated with a combination of antiviral antibodies and poly(I:C). These data confirm that antibody-mediated immunity plays a significant role in resistance against coxsackievirus B3 infection and suggest that antiviral antibodies may interact with poly(I:C) or work independently to produce an enhanced protective effect. PMID- 6288571 TI - Host plasma proteins on the surface of pathogenic Trichomonas vaginalis. AB - Sodium dodecyl sulfate-gel electrophoresis and fluorography and fluorography technology revealed that pathogenic Trichomonas vaginalis was able to acquire numerous loosely associated plasma proteins during incubation in normal human plasma. These proteins were readily removed by repeated washing of the parasite in phosphate-buffered saline. Plasma proteins avidly bound to the surface of T. vaginalis were also detected using a highly sensitive and specific agglutination assay with protein A-bearing Staphylococcus aureus pretreated with monospecific antiserum directed against individual human serum proteins. These avidly associated plasma proteins could not be removed by repeated washing in phosphate buffered saline or by treatment of washed, live organisms with surface-modifying reagents such as trypsin and periodate. A combined radioimmunoprecipitation-gel electrophoresis-fluorography methodology indicated that parasite biosynthesis of hostlike macromolecules was not responsible for the observed agglutination and reinforced the idea of trichosomal acquisition of plasma components. Finally, incubation of trichomonads with plasma in various buffers at different pH values did not alter the agglutination patterns. These and other data suggest that specific membrane sites trichomonal binding of host proteins. The biological significance of our results is discussed. PMID- 6288569 TI - Genetic relatedness among human rotaviruses as determined by RNA hybridization. AB - Viral RNAs from human rotaviruses were compared by gel electrophoresis and by hybridization to probes prepared by in vitro transcription of two well characterized laboratory strains (Wa and DS-1). Also, the viral RNAs were compared by hybridization to probes prepared from three of the test viruses. Thirteen specimens (diarrheal stools) were obtained from infants and children 5 to 21 months old on a single day at the emergency ward of the Caracas Children's Hospital, and an additional specimen was obtained from the same hospital 6 months before. When the electrophoresed viral RNAs were stained with ethidium bromide and examined by UV light, five different migration patterns (electropherotypes) were distinguished on the basis of differences in mobility of the RNA segments. The hybridization technique that was employed permitted only qualitative comparisons of corresponding genes of different human rotaviruses. Ten of the specimens contained enough virus to yield sufficient RNA for hybridization studies. Eight of the viruses studied by hybridization contained 4 to 11 genes that reacted specifically with the Wa probe to yield double-stranded RNA segments with a mobility similar to that of Wa viral RNA or test virus RNA. The other two viruses contained 11 genes that reacted specifically with the DS-1 hybridization probe to yield double-stranded RNA segments with a mobility similar to DS-1 viral RNA or test virus RNA. A more complex picture emerged when hybridization probes were prepared from three of the test viruses and used to compare the different electropherotypes. Corresponding genes that exhibited similar migration did not necessarily exhibit homology when studied by hybridization. Also, some corresponding genes that exhibited homology did not have the same mobility by gel electrophoresis. PMID- 6288572 TI - Effect of steroid hormones on virus-induced diabetes mellitus. AB - Female ICR Swiss mice, generally resistant to the diabetogenic effects of the D variant of encephalomyocarditis virus, develop diabetes to the same extent as males if they are pretreated with testosterone. The data suggest that testosterone is one of the factors involved in the development of diabetes in certain strains of mice. PMID- 6288573 TI - Comparison of simian and human cytomegalovirus reactivities in an enzyme-linked immunospecific assay: effect of antigen preparation on cross-reactive antigens. AB - Simian cytomegalovirus was substituted for human cytomegalovirus in an enzyme linked immunoassay. Unlike the indirect immunofluorescence assay which demonstrates a two-way cross-reactivity, only one-way cross-reactivity was observed. Altering the method of simian antigen preparation gave some insight other this different reactivity. PMID- 6288574 TI - Partial characterization of human spermatozoal phosphodiesterase and adenylate cyclase and the effect of steroids on their activities. AB - In sonicated human spermatozoa, phosphodiesterase hydrolyzed adenosine 3':5' monophosphate (cAMP) at 20.80 +/- 3.23 nmoles/10(8)sperm/20 min at 37 degrees C (50 microM cAMP, initially). In human semen, about 60% of the phosphodiesterase activity was in the spermatozoa. Both polyacrylamide gel electrophoresis and DEAE cellulose column chromatography indicated that there are at least 5 isozymes of phosphodiesterase. Various steroids were tested at a concentration of 2 micrograms/ml for their effects on phosphodiesterase activity in semen. None was found to have any significant effect. In sonicated human spermatozoa, adenylate cyclase synthesized cAMP at 0.02-2.11 nmoles/10(8)sperm/20 min at 37 degrees C (1 mM ATP, initially) depending on the availability of Mn2+ and caffeine in the assay mixture. Mn2+ was demonstrated to be a potent adenylate cyclase activator in human spermatozoa and its effect on human sperm adenylate cyclase was found to be dose-dependent. Cholera toxin, at a concentration of 20 micrograms/ml, had no effect on human sperm adenylate cyclase activity after it had been incubated with the intact spermatozoa for between 5 min and 5 h at 37 degrees C before the sperm were homogenized and the rate of cAMP formation assayed. In addition, human sperm adenylate cyclase decayed rapidly at 37 degrees C. Of various steroids tested at a concentration of 2 micrograms/ml for their effects on human sperm adenylate cyclase activity, only oestradiol-17 beta showed a significant effect, elevating the rate of cAMP formation by about 30%. PMID- 6288575 TI - Interaction of Epstein-Barr virus with leukaemic B cells in vitro. II. Cell line establishment from prolymphocytic leukaemia and from Waldenstrom's macroglobulinaemia. AB - Leukaemic B-cell populations were prepared from six patients with high-count prolymphocytic leukaemia (PLL) as well as from one patient with Waldenstrom's macroglobulinaemia (WM) in frankly leukaemic phase, and their response to in vitro Epstein-Barr (EB) virus infection was monitored in terms of expression of the virus-associated nuclear antigen EBNA and of virus-induced transformation to continuous cell lines. The individual leukaemic populations, tested on several occasions, gave reproducibly different responses one from another which were not obviously related to differences either of surface immunoglobulin phenotype or of immunoglobulin secretor status in vivo. After infection, four out of six PLL populations showed either transient or a more persistent expression of EBNA, always involving a minority of the cells, with no evidence of any virus-induced transformation up to six weeks. In contrast, two out of six PLL samples as well as the WM sample rapidly gave rise to EBNA-positive cell lines which, on the evidence both of restricted immunoglobulin class expression and of abnormal marker chromosomes, were clearly derived from the leukaemic cells. Further comparative studies of such leukaemic B-cell populations may help to define host cell components necessary for the triggering of EB-virus-induced cellular transformation. PMID- 6288576 TI - Mammary tumor induction in inbred mouse strains with urethane is not accompanied by changes in expression of B- and C-type retroviral structural proteins. AB - Urethane administrated in the drinking water of breeding female mice of five inbred strains (DDf, KF, C3Hf, ddY and C57BL) leads to a higher incidence of mammary tumors at an extremely early age in four of these strains (DDf, KF, C3Hf and ddY). In the case of the DDf strain the tumors are adenoacanthomas, but in the case of the other strains adenocarcinomas are also observed. The effect of urethane in the drinking water on expression of endogenous retroviral structural proteins of the B and C type (MTV and MuLV) was studied in the milk by an immunodiffusion test and in organ extracts by radioimmunoassays (for MTVp27, MTVgp52 and MuLVp30). Organs tested include salivary glands, thymus and spleen from female and male animals. In addition, prostate and seminal vesicles were tested in male animals and uterus and mammary glands in females. Mammary tumors induced by urethane were also treated. Urethane does not induce or enhance endogenous retroviral antigen expression, either in these organs or in the mammary gland, one of the target organs for tumorigenesis by this agent. PMID- 6288577 TI - Chromosomes of human hepatoma cell lines. AB - The karyotypes of three human hepatoma cell lines Hep G2, Hep 3B and PLC/PRF/5 were investigated by G- and C-banding techniques. In addition to ploidy changes, typical for most carcinoma cell lines, certain markers were found that remained stable throughout passage of these cultures. Chromosome I is involved in multiple translocations, resulting in at least three copies of the chromosome I heterochromatin region in each cell line. Inversion in the 9qh region is also seen. In addition, each of the cell lines consistently contains trisomy of 17q. The rearrangements of chromosome I are most striking in the Hep 3B and PLC/PRF/5 cell lines, which are derived from human hepatocellular carcinomas and contain integrated copies of the hepatitis B viral genome. These two cell lines are characterized by the presence of at least five copies of the I (p13 leads to q21) region that result from multiple deletions and/or translocations; by consistent trisomy and polymorphism of the 9qh region; and by trisomy of chromosome 10 (also involved in rearrangements). The Hep G2 and Hep 3B cell lines behave functionally as highly differentiated liver parenchymal cells and are karyologically distinguishable from PLC/PRF/5 both by the presence of trisomy of 6 (pter leads to q14) and by the finding that one of the homologues of chromosome 15 is 15q+. PMID- 6288578 TI - Histopathological features of rapidly progressing breast carcinoma in Tunisia: a study of 94 cases. AB - The histological features of 94 cases of carcinoma of the breast seen in Tunisia were recorded and subsequently correlated with the clinical classification of the patients in terms of poussee evolutive (PEV) categories. Histological features analyzed in the breast tissues and skin included tumor type, nuclear grade, number of mitoses, involvement of the dermis, cutaneous inflammatory infiltrate and edema. Twenty-eight percent of the cases in which the skin was examined showed tumor emboli in the lymphatics of the dermis. The frequency of cutaneous permeation correlated with the PEV categories. The percentage of cases with nuclear grade 3 was higher for the group with rapid progression of the disease (PEV 1,2,3) than for the cases belonging to the PEV-0 category (90% versus 64.2%). The present study demonstrates that a large proportion of breast carcinoma patients in Tunisia suffer from "inflammatory carcinoma of the breast". PMID- 6288579 TI - Hepatitis B virus and primary hepatocellular carcinoma: family studies in Korea. AB - Chronic infection with hepatitis B virus (HBV) is closely associated with the etio-pathogenesis of primary hepatocellular carcinoma (PHC). It has been proposed that infection with HBV early in life, frequently transmitted by an HBV-carrier mother, leads to persistent infection with HBV, which in turn is associated with the development of chronic active hepatitis (CAH), post-necrotic cirrhosis and PHC. If this view is correct, then there should be clustering of chronic carriers of HBV in families of patients with chronic liver disease. We tested this hypothesis in Korea by collecting serum from 132 patients with these chronic liver diseases admitted to the Seoul National University Hospital and 664 of their first-degree relatives. Controls (636) were members of two churches in Seoul and a rural village population; 261 of the controls were between the ages of 30 and 59, the age range that included 95% of the cases of chronic liver disease. Sera were assayed for hepatitis B surface antigen (HBsAg), antibody to HBsAg (anti-HBs) and antibody to hepatitis B core antigen (anti-HBc). Almost all cases showed evidence of present or past infection with HBV; 80% were HBsAg(+) and 14% were anti-HBs(+); 100% of 47 cases of PHC, 100% of 35 cases of cirrhosis, and 94% of 50 cases of CAH were anti-HBc(+); 6% of males and 4% of females in control population (30-59 years of age) were HBsAg(+), 71% were anti-HBc(+), and 51% were anti-HBs(+). HBsAg(+) patients with chronic liver disease tended to be younger than HBsAg(-) patients with anti-HBs or anti-HBc antibodies. Mothers of patients with more frequently (HBsAg(+) (9 of 33) than age-matched women in the control population (0 of 34) or wives of patients (5 of 68). Five of 23 fathers were also HBsAg(+) compared with 1 of 25 age-matched controls. As first observed in Africa, there was a deficit of anti-HBs in the fathers of cases compared with the controls. Siblings of patients were frequently HBsAg(+) (45% of 154), with the highest prevalence in brothers (53%). Family history shows that five fathers, two mothers and five brothers of cases have died of PHC. These data are compatible with the hypothesis tested and lend further support to the view that prevention of infection with HBV will lead to a marked decrease in the incidence of CAH, cirrhosis and PHC in areas where these diseases are endemic. Members of the families of patients with these diseases are at high risk of developing persistent infection with HBV and chronic liver disease. It would be appropriate to focus preventive strategies on infants and children in such families. PMID- 6288580 TI - Detection of polyoma-virus-induced tumor antigen(s) by macrophage migration inhibition. PMID- 6288581 TI - Identification of the RPMI 8226 retrovirus and its dissemination as a significant contaminant of some widely used human and marmoset cell lines. AB - A retrovirus designated RPMI 8226V, isolated in 1973 from the human myeloma cell line RPMI 8226 has been characterized by competition radioimmunoassay (RIA) for the major viral structural protein and by nucleic acid hybridization analysis using cDNA of the virus. The virus is highly related to the squirrel monkey type D retrovirus, SMRV. In the homologous RIA using rabbit anti-RPMI 8226V and 125I labelled p37 of RPMI 8226V, RPMI 8226V and SMRV exhibited competition of 81% and 73% respectively. Similarly, in the homologous system for SMRV p36, these viruses competed 98 and 100%. Reagents made from the type D retrovirus. Mason Pfizer Monkey Virus (MPMV), known to be related but distinct from SMRV, were used in assays designed to detect interspecies determinants of type D retroviruses. In assays using goat anti-MPMVp26 vs SMRV 125I-p36, RPMI 8226V, SMRV and MPMV competed to the same extent (93%). Hybridization analysis of RPMI 8226V cDNA showed significant homology to cellular RNA and DNA of mink, bat, and human cell infected with RPMI 8226V and to DNA or SMRV infected cells but not to uninfected cells or cells infected with other viruses. These results taken together clearly indicate that RPMI 8226V and SMRV are very closely related to each other. The finding of a type D retrovirus in this human myeloma cell line that had been used in EBV studies (the usual source of EBV being the marmoset cell line B95-8) prompted a survey of RPMI 8226V in some human and marmoset cell lines. The assays included the RIA for p36, nucleic acid hybridization using cDNA of RPMI 8226V, reverse transcriptase analysis and electron microscopy (EM). The results clearly show that in addition to RPMI 8226, human Burkitt lymphoma cells BJAB/B-95-8/K which were supertransformed by EBV from B-95-8/K marmoset cells as well as marmoset cell lines [(B-95-8/K and B-95-8/N) obtained from Stockholm and Uppsala, Sweden] were positive for the RPMI 8226V. Similar lines obtained elsewhere were negative. The results obtained clearly indicate that RPMI 8226V is a serious laboratory contamination in some widely used human cell lines. The possible impact of this viral contamination for some virological and cell biological studies is discussed. PMID- 6288582 TI - Effects of pituitary peptides on fat mobilisation. AB - No reliable answer can yet be given whether the pituitary gland is physiologically involved in the process of fat mobilization. The reasons are, for example, different test systems and criteria of lipolytic activity as well as variable criteria of purity of the peptides tested. Under comparable test conditions MSH, beta-endorphin, ACTH and beta-lipotropin turned out to be the most lipolytically active peptides. There is also increasing evidence that these peptides are involved in metabolic processes especially in glucose metabolism. The lipolytic activity of beta-lipoprotein, ACTH and MSH is mediated by a common heptapeptide (beta-LPH 47-53) furthermore, beta-lipotropin has a second message sequence in the C-terminal part of its molecule (beta-endorphin). This second message sequence is located in the amino acid sequence beta-LPH 77-86. The lipolytic effect of all these peptides is accompanied by an activation of adenylate cyclase. The effect of beta-endorphin is not mediated by an opiate receptor. The further elucidation of the lipolytic activity of these peptides investigations were necessary on the degradation of the peptides at the site of action as well as the use of test systems nearer to physiological conditions for example perifused fat cells. In conclusion, there are condensing data on the possible physiological role of brain and pituitary peptides in metabolic regulation. PMID- 6288583 TI - Dehydro-enkephalins. III. Synthesis and biological activity of [delta Ala2, Leu5] enkephalin. AB - [delta Ala2, Leu5]-enkephalin has been prepared and shown to be more active than the parent saturated enkephalin in a binding assay using rat brain membranes and [3H]dihydromorphine as a tracer. In a comparison of potencies against [3H]dihydromorphine and [3H]-[D-Ala2, D-Leu5]-enkephalin as tracers, [delta Ala2, Leu5]-enkephalin showed preference for micro opiate receptors, possibly due to the hydrophobicity of the delta Ala2 residue. A synthetic tetrapeptide enkephalin [delta Ala2]-desLeu5-enkephalin had weak activity and high selectivity for the micro receptors. O-Acylation of a serine residue in the peptide was achieved by coupling between the peptide and a carboxylic acid using DCC and a catalytic amount of 4-dimethylaminopyridine. PMID- 6288584 TI - Biological and immunological characterization of iodinated bovine growth hormone. AB - Iodinated bovine growth hormone, containing no more than 1 g-atom of iodine per mole of hormone is generally used as a tracer in studies related to the action and metabolism of the hormone. This derivative was tested in different biological and immunological systems in which the hormone is known to be active. The iodinated derivative was almost indistinguishable in its properties from the native hormone when it was examined by the following criteria: body growth promoting activity, rat liver uptake in vivo, binding to rabbit liver microsomes and primary antigen-antibody interactions. Micro-complement fixation experiments suggested that the iodination produces minor alterations in the affinity of some antigenic determinants. PMID- 6288585 TI - Human somatotropin 61. A simple method to predict potential of somatotropin fragments for noncovalent recombination by radioreceptor assay. AB - Different mixtures of reduced-alkylated thrombin fragments of human and sheep somatotropin have been tested for binding affinity to liver membranes. The bioassay data correlated well with the abilities of the fragments to form noncovalent recombinants as shown in separate biochemical studies. PMID- 6288586 TI - Beta-Endorphin. Synthesis and radioreceptor-binding activity of the ostrich hormone. AB - Ostrich beta-endorphin has been synthesized by the solid-phase method. Opiate activity in a radioreceptor binding assay is about seven times that of human beta endorphin. Structural differences within positions 6-15 account for the increased binding potency. PMID- 6288587 TI - Beta-Endorphin. Opiate receptor-binding activity of synthetic analogs modified in the enkephalin segment in rat brain membrane and neuroblastoma x glioma hybrid cell. AB - The potency of a series of synthetic analogs of beta-endorphin in inhibiting binding of [3H2-Tyr27]-beta h-endorphin to either rat brain membranes or neuroblastoma x glioma hybrid cells (NG108-15) has been determined and compared with the previously determined analgesic potency. There is a very good correlation between inhibitory potency in membranes and cells, but the correlation between analgesic potency and inhibitory potency in either membranes or cells is not as good. PMID- 6288589 TI - Synthesis and properties of human beta-endorphin-(1-9) and its analogs. AB - Four analogs of human beta-endorphin (beta h-EP) were synthesized by the solid phase method: beta h-EP-(1-9) (I), [D-Ala2]-beta h-EP-(1-9) (II), [Gln8]-beta h EP-(1-9) (III), and [D-Ala2, Gln8]-beta h-EP-(1-9) (IV). Measurement in a radioreceptor binding assay with use of tritiated beta h-EP as primary ligand gave relative potencies as follows: Met-enkephalin, 100; I, 76; II, 100; III, 200; IV, 200. Two new amino acid derivatives were prepared and used for synthesis of the analogs: N alpha-t-butyloxycarbonyl-O-(cyclopentyl)-tyrosine and N alpha-t butyloxycarbonyl-gamma-(cyclopentyl)-glutamic acid. PMID- 6288588 TI - Beta-Endorphin. Interaction of synthetic analogs having different chain lengths with morphine and enkephalin receptors in rat brain membranes. AB - Human beta-endorphin analogs with various chain lengths have been investigated for their potency in displacing tritiated dihydromorphine and Leu-enkephalin binding in rat brain membrane preparations. It was found that the reduction of chain length from residues 1-31 to 1-5 led to a gradual loss of preference for the morphined receptor. In addition, the extension of the chain length of the Met ekephalin segment to the COOH-terminal glutamic acid modified the binding of the NH2-terminal sequence to the enkephalin receptor. The fact that camel beta endorphin is more potent in displacing the two tritiated primary ligands than the human hormone is also reported herein. PMID- 6288590 TI - A functioning complex of two cytochrome c fragments with deletion of the (39-58) eicosapeptide. AB - Two major fragments of horse heart cytochrome c involving the sequences (1-38) and (60-104) were found to produce a stable complex. The two fragments were devoid of any cytochrome c activity. The complex exhibited a hardly measurable electron transfer capacity with respect to cytochrome c oxidase and missed the 695 nm absorption band. The introduction of tryptophan in position 59 restored the intrinsic activity of the complex to the level of native cytochrome c. This was concluded from the convergence of the Eady-Hofstee plots which extrapolate to the same Vmax at high substrate concentrations. The absorption spectrum of the complex in the ferriform contained a clear absorption band at 695 nm (84% of that found with native cytochrome c). The investigation proves the indispensability of tryptophan in position 59 for the transfer of an electron to cytochrome c oxidase and supports the conclusions of Parr et al. about the existence of two consecutive processes in the folding of the two fragments (vide infra). PMID- 6288591 TI - Comparison of the thermal stability characteristics of native and deglycosylated ovine pituitary lutropin. AB - The receptor binding, immunological and biological activities of native ovine lutropin were almost completely eliminated when aqueous solutions of the hormone were kept in a boiling water bath for 30 or 60 min. Similar exposure of chemically deglycosylated lutropin revealed that this preparation was relatively more stable to heat treatment. The conformational features of deglycosylated lutropin required for receptor binding and immunological activity were significantly retained after thermal treatment. The heated deglycosylated lutropin solution still retained its ability to antagonize cyclic AMP accumulation stimulated by the native hormone in rat testicular interstitial cells. Specificity of receptor (lutropin) binding or inhibitory activity was not lost by heating of deglycosylated lutropin as revealed by lack of an effect in follitropin radio-receptor assays. PMID- 6288592 TI - Isolation and properties of beta-endorphin-(1-27)-like peptide from bovine brains. AB - A beta-endorphin-(1-27)-like peptide was isolated from bovine cerebral hemisphere extracts by gel filtration, ion-exchange chromatography, high performance liquid chromatography and paper electrophoresis. The peptide had tyrosine as the amino terminal residue and its amino acid composition was nearly identical to that of equine pituitary beta-EP-(1-27). It had also the same mobility as equine pituitary beta-EP-(1-27) in paper electrophoresis. In radioimmunoassay and opiate receptor-binding assay, the brain peptide had 50% activity when compared with human beta-endorphin-(1-27). Evidence for the occurrence of NH2-acetylated form of beta-endorphin-(1-27) is also presented. PMID- 6288593 TI - Beta-Endorphin. Synthesis and properties of double-headed and related analogs. AB - A double-headed analog of human beta-endorphin (betah-EP), N,N'-bis (beta endorphinyl)-cystine (II), has been synthesized by the solid-phase method, along with betah-EP-Cys(CH2CONH2)-OH (I) and [Tyr31]-betah-EP (III). Their relative potencies in a radioreceptor-binding assay were: betah-EP, 100; II, 235; I, 170; and III, 204. In the tail-flick test for analgesic activity their relative potencies were: betah-EP, 100; II, 86; I, 93; and III, 116. PMID- 6288594 TI - Melanotropins: aldosterone- and corticosterone-stimulating activity in isolated rat adrenal cells. AB - Stimulation of corticosterone and aldosterone production in rat adrenal decapsular and capsular cells respectively by alpha- and beta-melanotropins (alpha- and beta-MSH) have been investigated. Although they are considerably less potent than corticotropin (ACTH), the dose-response curves are parallel to those for ACTH except in the case of aldosterone stimulation by beta-MSH where the dose response curve is biphasic. The steroidogenic actions of ACTH, beta-MSH and alpha MSH are antagonized by corticotropin-inhibiting peptide in both capsular and decapsular cells. A synthetic analog of beta-MSH ([Gly10]-betac1-MSH) inhibits both beta-MSH and ACTH in aldosterone output in capsular cells, but it does not inhibit their corticosteroidogenic actions in decapsular cells. PMID- 6288595 TI - Synthesis and properties of human beta-endorphin-(1-17) and its analogs. AB - Four analogs of human beta-endorphin (beta h-EP) were synthesized by the solid phase method: beta h-EP-(1-17) (I), [D-Ala2]beta h-EP-(1-17) (II), [Gln8]-beta h EP-(1-17) (III) and [D-Ala2, Gln8]-beta h-EP-(1-17) (IV). Measurement in a radio receptor binding assay with use of tritiated beta h-EP as primary ligand gave relative potencies as follows: Met-enkephalin, 100; I, 33; II, 47; III, 889; IV, 123; beta h-endorphin, 2253. PMID- 6288596 TI - beta-Endorphin: analgesic and receptor binding activity of non-mammalian homologs. AB - Analgesic potencies of turkey, ostrich and des-acetyl salmon beta-endorphins have been measured in the tail-flick test and binding affinities determined by radio receptor assay. The duration of analgesia and the slope of the dose-response curves generated by these peptides are similar to those elicited by mammalian beta-endorphins. This suggests that they act in vivo and in vitro on the same population of opiate receptors. The ratio of binding to analgesic potencies observed for these peptides varies nearly sixfold. Structure-activity analysis suggests that a basic side-chain at position 9 is required in order to produce a high opiate activity both in vivo and in vitro. A reexamination of the biological activities of camel beta-endorphin shows that the analgesic potency and binding affinity of this peptide are respectively 171 and 2.7 times higher than human beta-endorphin. His-27 and/or Gln-31 may contribute to this increased potency. The dissociation of radioreceptor binding affinity from analgesic potency in these naturally occurring beta-endorphin homologs suggests that either the conditions under which the binding assay is performed mask the true binding potency in the brain or that, once bound to the appropriate receptor, these homologs do not possess equal ability to produce biological effects. PMID- 6288597 TI - Corticotropin inhibiting peptide and a synthetic beta-melanotropin analog inhabit the lipolytic activity of corticotropin and melanotropins. AB - Lipolysis in isolated rabbit fat cells induced by beta-melanotropin, alpha melanotropin and corticotropin was inhibited by both corticotropin inhibiting peptide and [Gly10]-beta-melanotropin. Corticotropin inhibiting peptide was a more potent antagonist than [Gly10]-beta-melanotropin. PMID- 6288598 TI - beta-Endorphin: synthesis and properties of analogs modified in positions 8 and 31. AB - Four analogs of human beta-endorphin (beta h-EP) were synthesized by the solid phase method: [Gln8,31]-beta h-EP(I), [Arg8,Gln31]-beta h-EP(II), [Ala8,Gln31] beta h-EP (III), and [Val8, Gln31]-beta h-EP(IV). Radioreceptor binding assay with use of tritiated beta h-EP as primary ligand gave relative potencies as follows: beta h-EP, 100; I, 200;II, 150;III, 150;IV, 120. Relative potencies in an analgesic assay were: beta h-EP, 100; I,236;II, 254;III, 116;IV, 121. The side chain of Glu-8 in beta h-EP can be replaced by a variety of structures without diminishing biological activity. PMID- 6288599 TI - Human beta-endorphin. 270-MHz 1H-nuclear-magnetic-resonance study of glycyl residues in aqueous solution. AB - 1H spectra at 270 MHz of the beta h-endorphin glycyl residues in aqueous solution are reported. The chemical shifts, coupling constants and temperature coefficients are compared with those of the glycyl residues in Met-enkephalin and in a random coil model peptide. The local conformation of Tyr-Gly-Gly-Phe-segment observed in Met-enkephalin is maintained in beta h-endorphin. PMID- 6288600 TI - Physicochemical characteristics of homogeneous bovine lung angiotensin I converting enzyme. Comparison with human serum enzyme. AB - Angiotensin I-converting enzyme was purified to electrophoretic homogeneity (12 units/mg) from bovine lung tissue and from human serum using an affinity gel described previously (Harris et al., (1981) Anal. Biochem. 111, 227-234). The isoelectric point (4.5), molecular weight (145 000), S20,W (8.1), amino acid composition and carbohydrate content of the lung enzyme are all similar to the values obtained for the human serum enzyme. The NH2-terminus of the lung enzyme (Ala) is different from that of the serum enzyme (Tyr) but the COOH-terminal sequences are identical (-Leu-Ser-OH). Pure bovine lung enzyme was reduced and carboxyamidomethylated with iodo (14C1) acetamide to the extent predicted by the number of cysteine residues. Since no radioactivity was incorporated into denatured enzyme that was not reduced, all of the cysteine residues must be in the form of disulfide bonds. Reverse-phase HPLC was used to separate peptides obtained from the lung enzyme after degradation with either trypsin or cyanogen bromide. The number of peptides resolved (42 after trypsin, 31 after cyanogen bromide), were only 20% fewer than the number predicted from the amino acid analysis and therefore the possibility that the converting enzyme (a single polypeptide chain) might be a fused dimer is excluded. PMID- 6288601 TI - Synthesis of human corticotropinyl-thiolglycine and its specific conjugation to bovine serum albumin. AB - Human corticotropin containing a thiolglycine residue at the carboxyl terminal (I) was synthesized by the solid-phase method. The citraconyl derivative of peptide I was coupled to either a model tetrapeptide or bovine serum albumin by reaction with silver nitrate/N-hydroxysuccinimide in water. The extent of reaction with bovine serum albumin was determined by radioimmunoassay, and the peptide-protein conjugate was shown to possess 12% of the steroidogenic activity of porcine adrenocorticotropin in isolated rat adrenal cells. Peptide I and its conjugate with the model tetrapeptide were fully active in the same system. PMID- 6288603 TI - DNA double-strand breaks generated by the repair of X-ray damage in Chinese hamster cells. PMID- 6288602 TI - E.S.R. of spin-trapped radicals in gamma-irradiated polycrystalline amino acids. Chromatographic separation of radicals. AB - The free radicals produced by gamma-radiolysis of polycrystalline amino acids (L valine, L-leucine, L-isoleucine and L-proline) at room temperature in the absence of air were investigated by spin trapping with 2-methyl-2-nitrosopropane (MNP). The spin adducts produced by dissolving the irradiated solids in aqueous MNP solutions were separated by high-performance liquid chromatography and then identified by e.s.r. Deamination (ring-opening reaction for L-proline) was observed for all amino acid. For L-valine and L-leucine, H-abstraction from the tertiary carbon in the side chains occurred. For isoleucine, H-abstractions from the alpha-carbon of the amino acid and from a non-terminal carbon in the side chain were found. PMID- 6288606 TI - Rapid changes in phospholipid metabolism during secretion and receptor activation. PMID- 6288604 TI - Modulation of certain immunologic responses by vitamin C. III. Potentiation of in vitro and in vivo lymphocyte responses. PMID- 6288605 TI - Benzodiazepine receptors in the central nervous system. PMID- 6288607 TI - Fluctuation of Na and K currents in excitable membranes. PMID- 6288608 TI - Biochemical studies of the excitable membrane sodium channel. PMID- 6288609 TI - The plasma membrane as a regulatory site in growth and differentiation of neuroblastoma cells. PMID- 6288610 TI - Separated anterior pituitary cells and their response to hypophysiotropic hormones. PMID- 6288611 TI - The effect of severe zinc deficiency on the morphology of the rat retinal pigment epithelium. AB - Male Sprague-Dawley weanling rats maintained on a controlled dietary zinc intake had an accumulation of osmiophilic inclusion bodies in the retinal pigment epithelium (RPE) of the zinc-deficient group. At 7 weeks of zinc deficiency there were marked ultrastructural alterations of the RPE. In some instances a deepening of the basal infoldings of the cells of the RPE were observed. There was also vesiculation and degeneration of the photoreceptor outer segments. No changes were observed in the pair-fed, weight-restricted, and ad libitum-fed controls. The possible functions of zinc in the retina are discussed. PMID- 6288612 TI - Increased concentration of glucocorticoid receptors in rabbit iris--ciliary body compared to rabbit liver. AB - Dexamethasone binding to both rabbit iris-ciliary body and rabbit liver glucocorticoid receptors was compared by means of a batch assay in which the binding components were adsorbed to hydroxylapatite. The results showed that homogenates of both the rabbit iris-ciliary body and the rabbit liver contained a single class of high-affinity receptors. Binding affinity was virtually identical for the two receptors. A comparison of total binding sites in homogenates of the iris-ciliary body and the liver revealed that the iris-ciliary body glucocorticoid receptor was present in nearly twice the concentration of the hepatic receptor. PMID- 6288614 TI - Cytoplasmic inclusions in rabbit extraocular muscle. AB - Cytoplasmic inclusion bodies, similar to those previously described in abnormal and normal human extraocular muscle, were observed in the orbital surface layer of the superior rectus muscle in rabbit. These inclusion bodies are composed of a flocculent material of low density studded with granular foci of increased density. In sequential samples of serially reconstructed muscle fibers visualized by electron microscopy, cytoplasmic inclusion bodies were seen in 4.5% of 1187 samples through multiply innervated fibers that vary systematically in diameter along their length; inclusion bodies were also seen in 0.8% of 354 samples through multiple innervated fibers of constant diameter. Cytoplasmic inclusion bodies were not seen in 1838 samples through singly innervated fibers. These data suggest that such inclusion bodies may occur preferentially in multiply innervated fibers. The present findings are not compatible with previous suggestions that such cytoplasmic inclusion bodies may be indicative of a pathologic or aging process. These findings are consistent with previous suggestions that such inclusion bodies are to be considered as normal structures in extraocular muscle. PMID- 6288613 TI - Adrenergic influence on iris stromal pigmentation: evidence for alpha-adrenergic receptors. AB - Adrenergic influence on iris pigmentation in newborn pigmented rabbits was studied. Selective adrenergic antagonists were used topically to determine whether they could inhibit iris pigmentation. Unilateral, topical administration of an alpha-adrenergic antagonist (thymoxamine hydrochloride 1/2%) was not associated with iris hypochromia. Adrenergic influence on iris stromal melanogenesis appears to be mediated by alpha-adrenergic receptors. PMID- 6288615 TI - Portraits of viruses: foot-and-mouth disease virus. PMID- 6288616 TI - Infection of human peripheral blood mononuclear cells by varicella-zoster virus. AB - The infection of human peripheral blood leukocytes by varicella-zoster virus (VZV) was studied using an infectious center assay, indirect immunofluorescence and electron microscopy. Subsets of freshly isolated leukocytes were prepared, including granulocytes, mononuclear cells from Ficoll-Hypaque gradients, lymphocytes, and glass-adherent monocytes. When each of these populations was inoculated with VZV (MOI = 0.1), there was no evidence of effective infection. However, when monocytes were cultured in vitro for 7 days, they differentiated into macrophages that were productively infected with VZV. Peak percentages of infectious macrophages were detected 8-24 h after inoculation (mean 17.5%; range 10.2-30.4%). Using indirect immunofluorescence, viral antigens were detected in the cytoplasm and at the nuclear membranes of infected macrophages between 24 and 72 h after infection. Electron microscopy demonstrated the appearance of viral particles in the nucleus by 24 h. Large numbers of virions, often collected in tubules or vacuoles, were present in the cytoplasm at 48 h. The difference between the infection of fresh monocytes and cultured macrophages by VZV might reflect differences in their metabolic or differentiation state. The possible significance of these observations to VZV infection of immunocompromised hosts is discussed. PMID- 6288617 TI - Complementation between infecting Epstein-Barr virus and intrinsic viral genomes in human lymphoid cell lines. AB - 13 human lymphoid cell lines previously characterized in terms of their intrinsic Epstein-Barr virus (EBV) genome content were infected or superinfected with EBV from the supernatant of P3HR-1 cells. The cell lines included 2 lymphoma cell lines that contain no detectable EBV genomes and 11 cell lines containing intrinsic EBV genomes. All the cell lines responded to EBV infection by expressing early antigen (EA). The number of EA-positive cells in the cultures was proportional to the viral concentration used for infection or superinfection. However, cell lines containing multiple intrinsic EBV genomes expressed higher amounts of EA-positive cells. Evaluation of EA expression versus the number of intrinsic EBV genome copies per cell in each line revealed EA to increase as the intrinsic EBV genome content of the cell lines increased in a nonlinear curve that was described by an exponential logistic function. The coefficient of determination R2 for this curve was greater than 0.9 for multiple experiments. The data suggest that 80-90% of the virus in P3HR-1 is defective and requires intrinsic viral genomes for expression of EA. Transactivation or complementation between intrinsic EBV genomes and infecting virus is supported by these observations. PMID- 6288618 TI - Simple assay method for susceptibility of human lymphocytes to Epstein-Barr virus infection. AB - On the basis of the sequential early events induced by Epstein-Barr virus (EBV), the infectivity of human lymphocytes to EBV was quantified as a plateau value of the maximum nuclear antigen (EBNA) synthesis preceding cellular DNA synthesis. The frequency of EBV-infectible cells was an average of 7.6% of unfractionated umbilical cord lymphocytes and was 4.9% in the counterparts from human adults. In the B-cell-enriched populations, the average plateau values were 29.2% in umbilical cord and 15.5% in adult samples. This method is simple and applicable for quantitative assay of the sensitivity to EBV infection of a given lymphocyte preparation. PMID- 6288619 TI - Stability of the cloned 'joint region' of herpes simplex virus DNA. AB - To isolate stable recombinants containing the 'joint region', or L-S junction, of herpes simplex virus DNA, the EcoRI restriction enzyme cleavage fragments were cloned into both coliphage lambda and plasmid vectors. The authentic joint region was found in the plasmid but not in the lambda vector. The plasmid-joint region recombinant DNAs appeared stable on limited passage. Subcloning the small BamHI L S junction fragment into plasmid pBR322 gave rise to both stable and unstable recombinant DNAs. PMID- 6288620 TI - Viral antibodies in multiple sclerosis. PMID- 6288621 TI - Enzyme immunoassay for cytidine 3',5'-cyclic monophosphate (cyclic CMP). AB - An enzyme immunoassay for cytidine 3',5'-cyclic monophosphate (cyclic CMP) is presented. This assay is based upon the principles of competitive reaction and the double antibody solid phase method. Succinyl cyclic CMP-human serum albumin conjugate was injected into rabbits. Specific anti-cyclic CMP antibodies were incubated with a mixture of succinyl cyclic CMP labeled with beta-D-galactosidase and standard or sample cyclic CMP that had been succinylated prior to assay. The antibody-bound beta-D-galactosidase-cyclic CMP conjugate was separated from that of free with a second antibody, anti-rabbit immunoglobulin G, that was immobilized to a polystyrene ball. Then, activity of the enzyme on the solid phase was fluorometrically determined. When cyclic CMP contents in biological materials were estimated, acid extracts were partially purified by Dowex 1 x 8 formate column chromatography. The present immunoassay allows the detection of as little as 0.5 fmol of cyclic CMP with practically no interference from other cyclic nucleotides and cytidine analogs. By use of the enzyme immunoassay technique, we determined the amounts of cyclic CMP in various tissues of rats. They were found to be as little as 0.24-0.51 pmol/g wet weight, which was roughly 3000 to 20,000 and 100 to 500 times less than those of cyclic AMP and cyclic GMP, respectively. PMID- 6288622 TI - The low density lipoprotein receptor pathway in animals and man. PMID- 6288623 TI - [Nutrient and fiber content of infant food]. AB - The food given to 50 infants growing up in their family and 23 infants nursed in creches was chemically analyzed in order to ascertain the nutrient and dietary fibre contents. The babies' age was between 6 and 12 months. The determination of energy percentages revealed a clear shift in favour of carbohydrates. Based on 1000 calories, the infants received 7.5-8.5 g of dietary fibre per day. Sodium content was too high whereas the iron intake was definitely insufficient. The baby food prepared in the family only showed slight differences in comparison with the food administered in creches. PMID- 6288624 TI - The contribution made by cytochemistry to the study of the metabolism of the normal and rheumatoid synovial lining cell (synoviocyte). PMID- 6288625 TI - Electrophoretically separated bone cell types from the foetal rat calvarium: a histochemical and biochemical study. AB - Isolated cells obtained from foetal rat bone (calvarium) by collagenase digestion can be separated into three subpopulations on the basis of surface charge by free flow electrophoresis. These subpopulations have been tentatively identified by numerical, biochemical and functional criteria and are believed to be composed of: (1) bone resorbing cell types, designated Peak I cells; (2) fibroblasts and loose connective tissue cells, designated Peak II cells; and (3) a mixture of osteoblasts and osteoprogenitor cell types, designated Peak III cells. The anatomical position of these subpopulations in the whole calvarium was determined by comparing the results of histochemical and morphological experiments with the results of biochemical experiments. It was found that Peak I cells are located predominantly on the ventral (endocranial) surface, Peak II cells in the connective tissue periosteal membranes and Peak III cells on the dorsal (ectocranial) surface and in the suture line areas. The response of these cell types to parathyroid hormone and calcitonin with regard to c amp production and 45Ca release from devitalized bone is examined and indicates that cells from Peak I and Peak III both respond to parathyroid hormone but only the cells from Peak I respond to calcitonin. PMID- 6288627 TI - The effect of dietary fibre on D-xylose absorption. AB - Healthy probands ingested 25 g D-xylose dissolved in water or mixed with a test meal, with or without a supplement of guar or wheat bran. Viscosity increased to a maximum of 22,167mPa.s. Five or 10 g guar delayed xylose excretion only when administered in aqueous solution. When mixed with a test meal, guar accelerated xylose excretion. Bran had no effect on xylose excretion, independent of the manner of administration. Our results show that under these experimental conditions, the effect of guar on absorption is not dependent on the degree of viscosity. PMID- 6288626 TI - Pancreatic atrophy in copper-deficient rats: histochemical and ultrastructural evidence of a selective effect on acinar cells. AB - Copper deficiency had a differential effect between tissues in the rat pancreas. There was marked loss and atrophy of acinar cells, in which both hypertrophied and degenerating mitochondria were present. Cytochrome oxidase activity in acini was greatly depleted while monoamine oxidase activity was enhanced. Atrophy of acinar cells was accompanied by extensive degeneration of the rough endoplasmic reticulum, and by a failure of zymogen granule synthesis. These changes contrasted strongly with the appearance of non-acinar tissues, in which hypertrophy and degeneration of mitochondria were rarely observed. Islet tissue, pancreatic ducts and blood vessels showed no atrophic changes. Cytochrome oxidase activity in islet tissue, and in the epithelium of the main pancreatic ducts, appeared unaffected. PMID- 6288630 TI - Biological effects of slate dust: solubility and hemolytic studies. PMID- 6288629 TI - Effect of BCG on experimental asbestosis. PMID- 6288628 TI - Postnatal differentiation of sex-specific distribution patterns of G6Pase, G6PDH and ME in the rat liver. AB - The activity of the liver enzymes G6Pase, G6PDH and ME was studied in rats of 2-9 weeks old by histochemical means. In addition, G6PDH and ME activity was quantitatively determined in homogenates. In the 2nd and 3rd week G6Pase is similarly distributed in both sexes: while in the periportal zone high activity is demonstrable, the perivenous zone shows only low activity. After this period a nearly homogeneous distribution pattern becomes evident in all animals. Sex difference occurs after the 6th week: in the livers of male rats the periportal "maximum" is sometimes combined with a second peak in the perivenous area, in females a steep gradient emerges with high activity in the periportal zone and a low one in the perivenous zone. In the first postnatal weeks G6PDH activity is very low in parenchymal cells, but very prominent in Kupffer cells. Around the 5th week there is an increase, predominantly in the perivenous zone of both sexes. While there is again a further decrease demonstrable in male rats, the G6PDH activity of female rats rises to high adult values. This increase seems to be restricted to the perivenous zone. ME can be demonstrated at first in leucocytes. In the course of the 3rd week there is an increase of activity in both sexes: ME is demonstrable in parenchymal cells of the perivenous area and in scattered hepatocytes of the periportal area. In male rats, the perivenous activity is diminished towards the end of the investigation period, in females, however, a high activity remains in the perivenous zone. The data show that in females the activity of NADP dependent enzymes is high in the perivenous zone, so it may be assumed that a lipogenic area is situated around the terminal efferent vessels. Because of the sex difference this area may be hormone-dependent. The lipogenic area is situated opposite to the gluco(neo)genic area which corresponds to the periportal zone. PMID- 6288631 TI - [The assessment of the function of the nervus intermedius by means of functional salivary gland-scintigraphy]. AB - Using functional scintigraphy of the salivary glands, the function of the nervus intermedius can be assessed by estimating excretory quotients for both submandibular glands. This method is preferred to the standard salivation test method of Magielski and Blatt, despite a minimal exposure of the patient to radiation from the injected Natriumpertechnetat. The technical course of this investigation, along with the indications for its use, will be presented. PMID- 6288632 TI - Histocompatibility types and antiviral antibodies in a diverse group of individuals. AB - A study was made of the possible relationship between histocompatibility (HLA) antigens of the A and B loci and antibodies to cytomegalovirus, influenza A (victoria strain), measles, rotavirus, vaccinia, and varicella/zoster. A large and diverse group of unselected individuals was studied. A possible relationship was detected between the presence of the antigen B15 and a lack of circulating measles antibodies. The study group was divided into three sections: (a) healthy individuals, (b) "renal disease" patients awaiting renal transplantation, and (c) patients with various types of "other disease," mainly of an immunological nature. Significantly elevated titers to cytomegalovirus and varicella/zoster were found in the two diseased groups, but these elevated titers could not be linked with the presence or absence of any particular HLA A or B antigen. PMID- 6288635 TI - The use of acrylic resin oral prosthesis in radiation therapy of oral cavity and paranasal sinus cancer. AB - In radiation therapy of cancers of the oral cavity and the paranasal sinuses, the extent to which the tissues of the oral cavity are included in the radiation treatment portals will determine the severity of the oral discomfort during treatment. This will affect the nutritional status of the patients, and may eventually affect the total dose of radiation which the patients can receive for treatment of their cancers. In cooperation with the Maxillofacial Prosthetic Department, an acrylic resin oral prosthesis was developed. This prosthesis is easy to use and can be made for each individual patient within 24 hours. It allows for maximum sparing of the normal tissues in the oral cavity and can be modified for shielding of backscattered electrons from heavy metals in the teeth. We have also found that acrylic resin extensions can be built onto the posterior edge of post-maxillectomy obturators; this extension can be used as a carrier for radioactive sources to deliver radiation to deep seated tumor nodules in the paranasal sinuses. PMID- 6288633 TI - Intracranial tumors: response and resistance to therapeutic endeavors, 1970-1980. PMID- 6288634 TI - Internal mammary lymphoscintigraphy in the assessment of patients with ovarian carcinoma. AB - Radiocolloid internal mammary lymphoscintigraphy (IML) was evaluated in 364 patients with ovarian carcinoma to determine the frequency of abnormalities in post-operative patients, the association between the results of the lymphoscintigram and known clinical prognostic variables, and to establish whether IML yielded predictive information independent of these variables. Results of IML showed a correlation with established clinical prognostic features and yielded independent prognostic information. The sensitivity and specificity of IML in predicting relapse are 51% and 71% respectively, indicating that a single post-operative IML does not predict relapse or freedom from relapse with sufficient accuracy to make it a clinically useful test even though it provides an independent prediction of relapse. PMID- 6288636 TI - Lung inflation, lung solute permeability, and alveolar edema. AB - A series of experiments in anesthetized rabbits were conducted to determine whether hyperinflation of the lung alone could produce a protein-permeable lung epithelium and whether a protein-permeable lung epithelium allowed accumulation of liquid in the alveolar space. Some animals had their entire lungs subjected to distending pressures; others had only an area of the lung subjected to the high distending pressure. Alveolar liquid was measured by dilution of radioactive solutes upon instillation of saline into atelectatic lung, and protein permeability was determined by the loss of labeled albumin from the alveolar space over 40-60 min. Inflation of the entire lung at 40 cmH2O for 20 min increases air-space gas volume three- to fourfold, does not produce a protein permeable epithelium, and does not result in accumulation of alveolar liquid. Distension of a small area of the lung by 40 cmH2O pressure for 20 min increases the gas volume 6- to 12-fold and produces a protein-permeable epithelium, but does not result in liquid accumulation in the alveoli. It is concluded that only very high distending volumes cause the lung epithelium to become permeable to protein and that a protein-permeable epithelium alone does not induce alveolar edema. PMID- 6288637 TI - Neuromuscular adaptation in human thenar muscles following strength training and immobilization. AB - The effects of strength training and limb immobilization on the human thenar muscles were investigated in 11 healthy subjects. One group (n = 6) trained prior to immobilization and a second group (n = 5) underwent immobilization prior to training. Measurements made in the control condition and following the two experimental conditions included voluntary isometric strength, motor-unit counts, motor nerve conduction velocity, reflex potentiation, and isometric twitch contraction properties. When the results of both groups were combined an average of 5 wk of immobilization was found to cause a significant decrease in voluntary strength (42%, P less than 0.05) and reflex potentiation (37%, P less than 0.01) in relation to the control condition. Training caused an increase (40%, P less than 0.05) in voluntary strength and a decrease in twitch tension (25%, P less than 0.01) and contraction time (8%, P less than 0.05). Training prior to immobilization provided a reserve of neuromuscular function, which attenuated the effect of immobilization in relation to the control condition. It was concluded that neural as well as muscular adaptation occurred in response to immobilization. PMID- 6288638 TI - Fatigue of the mammalian diaphragm in vitro. AB - Diaphragm fatigue was studied in innervated diaphragm strips from 63 Sprague Dawley rats. The experiments examined 1) the effect on the rate of diaphragmatic fatigue of increases in the diaphragm's duty cycle, i.e., the ratio of the period of diaphragmatic contraction (Ti) to the duration of a cycle of contraction and rest (Ttot) and 2) the possibility that impaired neural transmission contributed to the fatigue process. Alterations in the duty cycle of the diaphragm were simulated by varying the pattern of electrical stimuli applied cyclically to the phrenic nerve. Fatigue was assessed from the rate of fall of isometric tension when the muscle was made to contract 90 times/min. The contribution of neural element fatigue was assessed by comparing the tension during phrenic nerve stimulation to the tension developed when the muscle was stimulated directly. Increasing the duty cycle (Ti/Ttot) from 25 to 50 to 75% increased the rate of diaphragmatic fatigue progressively. Holding Ti/Ttot constant at 75%, while varying Ti and Ttot, did not affect the rate of fatigue. Increases in duty cycle appear to increase the rate of fatigue by increasing the number of times the contractile process was activated. In fatigued muscle strips diaphragmatic tension was greater in directly stimulated muscle than in muscle strips activated via the phrenic nerve. The results indicate that 1) when the breathing action of the diaphragm is simulated in vitro, increases in duty cycle accelerate the fatigue process and 2) failure of transmission of phrenic impulses to diaphragmatic muscle cells contributes to the fall in tension during fatigue. PMID- 6288639 TI - Herpetic lymphangitis in a student population. PMID- 6288640 TI - Asian nursing students in the United States. AB - It is obvious that there is a great need to familiarize faculty and students with the Asian culture and heritage and to sensitize them to the difficulties and problems that Asian nursing students encounter in their adjustment to the university nursing program in the United States. Recommendations and strategies to achieve the above goals are: (1) Organizing cross-cultural courses for Asians and non-Asians to familiarize them with different cultures, (2) sensitizing faculty and counselors to the detrimental effects of existing nursing programs on international students, and (3) helping Asian nursing students better adjust to the American culture by providing English tutorial classes, support groups and host families that will act as socializing agents during the student's adjustment process. Through such educational and support programs, it is hoped that Asian nursing students will experience fewer difficulties which in turn will make their studies more meaningful and applicable. PMID- 6288641 TI - Latin American nursing students in the United States. AB - The Latino students confront a unique set of difficulties and patterns of adjustment in the majority culture and the educational experiences in the United States. Careful consideration must be given not only to the general characteristics of this group, but also to the individual's history of immigration, socioeconomic status, educational experiences and background. Multicultural education for the international nursing students can benefit all students, but it should not gloss over conflicts of values and beliefs. It must honestly deal with differences and similarities. It must be based on experience, not on stereotypes of root cultures or ethnic characteristics. In addition to addressing the cognitive affect and skill needs of all students, the multicultural education must address the special needs of minorities. Anglo students or majority-group students need realistic opportunities for experience and interaction with minority students and minority-faculty. Every group has something to teach the other. Faculty advisors and majority students should be encouraged to acquaint themselves with the need to establish rapport with the Latino student. There is also a need to develop in the classroom an atmosphere that enhances the learning experiences. The learning experiences should be so designed that all students and school personnel can intellectually, emotionally and socially participate on equal terms in a culturally diverse society. PMID- 6288642 TI - Middle Eastern nursing students in the United States. AB - The United States, then, surprises, upsets, delights, challenges, and to some extent, changes Middle Eastern students. For those who have studied in the United States and participated in our study, their experience has been rewarding in many respects and not only in academic terms. "The academic degree is only part of the experience of studying in America," says one Lebanese student, "One gains perspective, a global look and broadens horizons as one interacts and is being exposed to the society." Middle Eastern nursing students in the United States can perhaps look at their experiences more positively and the challenges of the academic setting more constructively if they have the support of faculty and classmates and if they are allowed the flexibility in structuring their own programs to meet their own and their countries' health needs. PMID- 6288643 TI - Canadian and European students in the United States. PMID- 6288644 TI - Nigeria: the land, its people, and health care. AB - The unique health care needs in developing countries and the responsibilities of the nursing profession in such countries place a heavy responsibility on schools of nursing involved in the education of foreign nursing students. The students in this study indicated that it was frustrating to repeat course content that was already familiar to them knowing that there are many courses that would better meet their needs. In developing countries, like Nigeria, it has been suggested that there is a need to prepare a "comprehensive nurse"--a combination bedside nurse, public health nurse, and midwife who can provide preventive and curative services in multiple settings. Such a task is awesome and illustrates the need for schools of nursing to provide an academic counselor who has some knowledge of the culture and health needs of the students' native land. When asked: "How could the school of nursing have helped you cope with your academic problems?" 43% of the students suggested a foreign student advisor or counselor. "To have a foreign student advisor who has been overseas and who will relate to the student at their level," replied one student. Only when education is relevant to the specific needs of a given country, will there be an improvement in the health of the people of that country. To educate registered nurses abroad is very costly for countries with low G.N.P. If we accept students from developing countries into our nursing programs, we have a responsibility to prepare them to be effective practitioners when they return to their homeland. The special needs of nursing students and the health care needs of the community for which they are trained must therefore be integrated into their training. PMID- 6288645 TI - Bridges or barriers to success: the nature of the student experiences in nursing. PMID- 6288647 TI - Surviving graduate nursing programs in the United States--a personal account of an Asian-American student. PMID- 6288646 TI - Applicability of nursing school programs in meeting foreign students' needs and expectations. PMID- 6288648 TI - Orienting nurses from other countries to graduate education in the United States. PMID- 6288649 TI - Mode of action of oxanosine, a novel nucleoside antibiotic. AB - Oxanosine, a novel nucleoside, inhibits the growth of Escherichia coli K-12 on peptone agar, but not on Nutrient agar. This antibiotic activity was found to be bacteriostatic and was antagonized by guanine, guanosine, and guanylic acid. The growth of leukemia L 1210 cell was also inhibited by oxanosine, and the inhibition was reversed by guanylic acid. Oxanosine was confirmed to be a competitive inhibitor of GMP synthetase (E.C. 6.3.5.2) and the Ki value was 7.4 X 10(-4) M. PMID- 6288650 TI - Metabolism of estrogens in the gastrointestinal tract of swine. II. Orally administered estradiol-17 beta-D-glucuronide. AB - Studies were conducted to determine the absorption and metabolic fate of orally administered 3H-estradiol-17 beta-glucuronide (3H-E2-G) in swine. Xylazine tranquilized female pigs (5 to 6 wk old) were given .04, .4 or 4 mumol 3H-E2-G via stomach tube, and blood samples were collected from previously implanted jugular cannulas for 12 or 72 h. The entire gastrointestinal tract was removed from gilts euthanatized 12 h post-treatment, and free and conjugated estrogens were isolated from plasma and intestinal chyme by diethyl ether extraction and adsorption to Amberlite XAD-2 resin columns. After preparative thin layer chromatography of the conjugate fractions, the conjugates were cleaved by enzyme hydrolysis, solvolysis or acid hydrolysis. The freed estrogens were identified by thin layer chromatography. Plasma radioactivity peaked between 6 and 8 h after administration of the conjugate. None of the radioactivity in plasma was ether extractable. There was evidence for a decrease in absorption rate of radioactive estrogen in the high dosage group. The pattern of metabolites and urinary excretion or orally administered 3H-E2-G was similar to that reported for 14C-E2, except for the greater proportion of polar metabolites and delayed absorption, probably reflecting the need for the conjugate to be hydrolyzed first. The greater proportion of polar metabolites found in this study may have been due to the longer treatment period rather than the administration of the conjugated form of estradiol. PMID- 6288651 TI - In-vitro evaluation of sulbactam, a penicillanic acid sulphone with beta lactamase inhibitory properties. PMID- 6288652 TI - Cefotiam concentrations in bile and in the wall of the gallbladder in patients with biliary disease. PMID- 6288653 TI - Reverse phase liquid chromatographic determination of vitamins D2 and D3 in milk. AB - A single column reverse phase high pressure liquid chromatographic method is described for the determination of vitamins D2 and D3 in fluid milk. Resolution of vitamin D2 from D3 is helpful for use as an internal standard. The method involves overnight saponification at room temperature, extraction of unsaponifiables, precipitation of cholesterol, and aluminum oxide column cleanup. Sample extracts were chromatographed under isocratic conditions on a 10 micron Vydac reverse phase column using acetonitrile-methanol (90 + 10) as the mobile phase. In addition, a MicroPak MCH-5 reverse phase column with acetonitrile as the mobile phase was used with an automatic system for one product type. Thirty samples each of homogenized (3.8% fat), low fat (2.0% fat), and skim (less than or equal to 0.5% fat) milk spiked with 200, 400, and 600 IU vitamin D/qt were analyzed. Coefficient of variation (CV) and percent recovery for each product type and each spike level of vitamins D2 and D3 were calculated from 10 replicate analyses. Vitamin D2 recoveries for all product types at the 3 fortification levels varied from 85.2 to 99.7%; vitamin D3 recoveries varied from 85.9 to 98.8%. The minimum detectable quantity of vitamin D in milk was 15 IU/qt. PMID- 6288654 TI - Broad-host-range IncP-4 plasmid R1162: effects of deletions and insertions on plasmid maintenance and host range. AB - R1162 is an 8.7-kilobase (kb) broad-host-range replicon encoding resistance to streptomycin and sulfa drugs. In vitro deletion of 1.8-kb DNA between coordinates 3.0 and 5.3 kb did not affect plasmid maintenance, but a Tn1 insertion at coordinate 6.3 kb led to a recessive defect in plasmid maintenance. The only cis acting region necessary for plasmid replication appears to lie between the Tn1 insertion at coordinate 6.3 kb and a second Tn1 insertion at coordinate 6.5 kb. All R1162 sequences between position 6.5 kb and the EcoRI site at coordinate 8.7/0 kb were dispensible for replication in Escherichia coli and Pseudomonas putida. Plasmids carrying insertions in a variety of restriction sites in an R1162::Tn1 derivative were unstable in P. putida but stable in E. coli. Tn5 insertions in R1162 showed a hot spot at coordinate 7.5 kb. A Tn5 insertion at coordinate 8.2 kb appeared to mark the 3' end of the streptomycin phosphotransferase coding sequence. All R1162::Tn5 derivatives showed specific instability in Pseudomonas strains but not in E. coli. The instability could be relieved by internal deletions of Tn5 sequences. In the haloaromatic-degrading Pseudomonas sp. strain B13, introduction of an unstable R1162::Tn5 plasmid led to loss of ability to utilize m-chlorobenzoate as a growth substrate. Our results showed that alteration of plasmid sequence organization in nonessential regions can result in restriction of plasmid host range. PMID- 6288655 TI - Reduction of exogenous cytochrome c by Neurospora crassa conidia: effects of superoxide dismutase and blue light. AB - The reduction of externally added cytochrome c by Neurospora crassa conidia was observed. The reduction was stimulated by antimycin A and suppressed partially by superoxide dismutase. When conidia were treated with diethyldithiocarbamate, which inactivated endogenous superoxide dismutase, the cytochrome c reduction was stimulated. Blue light also stimulated the cytochrome c reduction. Azide, which inhibits photochemical reactions mediated by flavins, suppressed the blue light effect. Superoxide dismutase partially suppressed the cytochrome c reduction in the light. The results suggest that O2(-) participates in the cytochrome c reduction by conidia and the flavins or flavoproteins are candidates for the receptor pigment of blue light to stimulate the cytochrome c reduction. It was also suggested that the redox component(s), which could directly transfer its reducing equivalents to exogenous cytochrome c, was present at the surface of conidia. PMID- 6288656 TI - Transformation of restriction endonuclease phenotype in Streptococcus pneumoniae. AB - The genetic basis of the unique restriction endonuclease DpnI, that cleaves only at a methylated sequence, 5'-GmeATC-3', and of the complementary endonuclease DpnII, which cleaves at the same sequence when it is not methylated, was investigated. Different strains of Streptococcus pneumoniae isolated from patients contained either DpnI (two isolates) or DpnII (six isolates). The latter strains also contained DNA methylated at the 5'-GATC-3' sequence. A restrictable bacteriophage, HB-3, was used to characterize the various strains and to select for transformants. One laboratory strain contained neither DpnI nor Dpn II. It was probably derived from a DpnI-containing strain, and its DNA was not methylated at 5'-GATC-3'. Cells of this strain were transformed to the DpnI restriction phenotype by DNA from a DpnI-containing strain and to the DpnII restriction phenotype by DNA from a DpnII-containing strain. Neither cross transformation, that is, transformation to one phenotype by DNA from a strain of the other phenotype, nor spontaneous conversion was observed. Extracts of transformants to the new restriction phenotype were shown to contain the corresponding endonuclease. PMID- 6288657 TI - Anaerobic incubation enhances the colony formation of a polA recB strain of Escherichia coli K-12. AB - Escherichia coli strain E247 (polA1 recB21) has reduced colony formation (even at the permissive temperature of 30 degrees C) because of a poor suppressor mutation (sup-126). The colony formation was enhanced in the absence of oxygen about 3 fold at 30 degrees C and 10(6)-fold at 43 degrees C, suggesting that a polA recB strain was inviable due to oxygen toxicity. Colony formation was also increased by incubation in an agar medium containing the reducing agent thioglycolate and incubation in the presence of chloroform-killed Saccharomyces cerevisiae pet+ cells, but not pet cells. Since the E247 strain viability was inversely dependent on the oxygen pressure and since the strain was more sensitive to superoxide radical than either the polA or the recB mutant, it seems likely that the polA and recB genes play a role in repairing DNA damage during respiration. PMID- 6288658 TI - Helper plasmid cloning in Streptococcus sanguis: cloning of a tetracycline resistance determinant from the Streptococcus mutans chromosome. AB - A model system for testing the helper plasmid cloning system of Gryczan et al. (Mol. Gen. Genet. 177:459-467, 1980) was devised for the Streptococcus sanguis (Challis) host-vector system. In this system, linearized pVA736 plasmid efficiently transformed an S. sanguis (Challis) host containing a homologous plasmid, pVA380-1, but did not transform a plasmidless host or a host containing a nonhomologous plasmid, pVA380. In addition, whereas monomeric circular pVA736 transformed a plasmidless host with two-hit kinetics, it transformed a pVA380-1 containing host with one-hit kinetics. This helper plasmid cloning system was used to isolate two HindIII fragments (5.0 megadaltons [Mdal] and 1.9 Mdal in size) from the chromosome of Streptococcus mutans V825 which conferred high-level tetracycline resistance. One tetracycline-resistant clone was examined and found to contain three plasmids which were sized and designated pVA868 (9.0 Mdal), pVA869 (9.5 Mdal), and pVA870 (9.8 Mdal). Results of Southern blot hybridization and restriction endonuclease digestion confirmed that all three chimeras were composed of two HindIII fragments of the S. mutans V825 chromosome, as well as a large portion, varying in size for each chimera, of the 2.8 Mdal cloning vector, pVA380-1. Incompatibility observed between pVA380-1 and each of the chimeras indicated that replication of the chimeras was governed by the pVA380-1 replicative origin. Southern blotting experiments revealed that the chimeras hybridized to Tn916, providing the first evidence that transposon-related genes of enteric streptococcal origin are disseminated among oral streptococci. PMID- 6288659 TI - Evidence for a messenger function of cyclic GMP during phosphodiesterase induction in Dictyostelium discoideum. AB - Chemotactic stimulation of vegetative or aggregative Dictyostelium discoideum cells induced a transient elevation of cyclic GMP levels. The addition of chemoattractants to postvegetative cells by pulsing induced phosphodiesterase activity. The following lines of evidence suggest a messenger function for cyclic GMP in the induction of phosphodiesterase: (i) Folic acid and cyclic AMP increased cyclic GMP levels and induced phosphodiesterase activity. (ii) Cyclic AMP induced both cyclic GMP accumulation and phosphodiesterase activity by binding to a rate receptor. (iii) The effects of chemical modification of cyclic AMP or folic acid on cyclic GMP accumulation and phosphodiesterase induction were closely correlated. (iv) A close correlation existed between the increase of cyclic GMP levels and the amount of phosphodiesterase induced, independent of the type of chemoattractant by which this cyclic GMP accumulation was produced. (v) Computer simulation of cyclic GMP binding to intracellular cyclic GMP-binding proteins indicates that half-maximal occupation by cyclic GMP required the same chemoattractant concentration as did half-maximal phosphodiesterase induction. PMID- 6288660 TI - Properties of acetate kinase isozymes and a branched-chain fatty acid kinase from a spirochete. AB - Spirochete MA-2, which is anaerobic, ferments glucose, forming acetate as a major product. The spirochete also ferments (but does not utilize as growth substrates) small amounts of l-leucine, l-isoleucine, and l-valine, forming the branched chain fatty acids isovalerate, 2-methylbutyrate, and isobutyrate, respectively, as end products. Energy generated through the fermentation of these amino acids is utilized to prolong cell survival under conditions of growth substrate starvation. A branched-chain fatty acid kinase and two acetate kinase isozymes were resolved from spirochete MA-2 cell extracts. Kinase activity was followed by measuring the formation of acyl phosphate from fatty acid and ATP. The branched chain fatty acid kinase was active with isobutyrate, 2-methylbutyrate, isovalerate, butyrate, valerate, or propionate as a substrate but not with acetate as a substrate. The acetate kinase isozymes were active with acetate and propionate as substrates but not with longer-chain fatty acids as substrates. The acetate kinase isozymes and the branched-chain fatty acid kinase differed in nucleoside triphosphate and cation specificities. Each acetate kinase isozyme had an apparent molecular weight of approximately 125,000, whereas the branched-chain fatty acid kinase had a molecular weight of approximately 76,000. These results show that spirochete MA-2 synthesizes a branched-chain fatty acid kinase specific for leucine, isoleucine, and valine fermentation. It is likely that a phosphate branched-chain amino acids is also synthesized by spirochete MA-2. Thus, in spirochete MA-2, physiological mechanisms have evolved which serve specifically to generate maintenance energy from branched-chain amino acids. PMID- 6288661 TI - Aerobic and anaerobic respiratory systems in Campylobacter fetus subsp. jejuni grown in atmospheres containing hydrogen. AB - Maximum growth of Campylobacter fetus subsp. jejuni, strain C-61, occurred when the cultures were incubated with shaking in atmospheres containing approximately 30% hydrogen, 5% oxygen, and 10% CO2. Suspensions of cells grown under these conditions consumed oxygen with formate as the substrate in the presence of 0.33 mM cyanide, which completely inhibited respiration with ascorbate-N,N,N',N' tetramethyl-p-phenylenediamine and with lactate. Spectroscopic evidence with intact cells suggested that a form of cytochrome c, reducible with formate but not with lactate or ascorbate-N,N,N',N'-tetramethyl-p-phenylenediamine, can be reoxidized by a cyanide-insensitive system. Analysis of membranes from the cells showed high- and low-potential forms of cytochrome c, cytochrome b, and various enzymes, including hydrogenase, formate dehydrogenase, and fumarate reductase. The predominant carbon monoxide-binding pigment appeared to be a form of cytochrome c, but the spectra also showed evidence of cytochrome o. The membrane cytochromes were reduced by hydrogen in the presence of 2-heptyl-4 hydroxyquinoline-N-oxide at concentrations which prevented the reduction of cytochrome c with succinate as the electron donor. Reoxidation of the substrate reduced cytochromes by oxygen was apparently mediated by cyanide-sensitive and cyanide-insensitive systems. The membranes also had hydrogen-fumarate oxidoreductase activity mediated by cytochrome b. We conclude that C. fetus jejuni has high- and low-potential forms of cytochrome which are associated with a complex terminal oxidase system. PMID- 6288662 TI - Action of restriction endonucleases on transforming DNA of Haemophilus influenzae. AB - Cleavage of DNA from Haemophilus influenzae with restriction endonucleases caused inactivation of transforming ability to an extent that depended on the genetic marker and the enzyme. The rate of inactivation, but not the final level of survival, depended on the concentration of enzyme in the restriction digest. In general, the greatest extent of inactivation of transforming activity was obtained with endonucleases that are known to produce the shortest fragments. We electrophoresed restriction digests of H. influenzae DNA in agarose gels and assayed transforming activity of DNA extracted from gel slices. In this way, we determined the lengths of restriction fragments that contain genetic markers of H. influenzae. For the marker that we studied most thoroughly (nov), the shortest restriction fragment that possessed detectable transforming activity was a 0.9 kilobase pair fragment produced by endonuclease R . PstI. The shortest marker bearing restriction fragment that retained substantial transforming activity (50% of value for undigested DNA) was a 2.1-kilobase pair EcoRI fragment bearing the kan marker. Among marker-bearing restriction fragments 1 to 4 kilobase pairs in length, survival of transforming activity varied 10,000-fold. We relate these observations to the recent findings by Sisco and Smith (Proc. Natl. Acad. Sci. U.S.A. 76:972-976, 1979) that efficient entry of DNA into competent H. influenzae cells appears to require the presence of a recognition sequence that is scattered throughout the Haemophilus genome in many more copies than in unrelated genomes. PMID- 6288663 TI - Antagonism of the B subunit of DNA gyrase eliminates plasmids pBR322 and pMG110 from Escherichia coli. AB - The constructed plasmid pBR322 and the native plasmid pMG110 were eliminated (cured) from growing Escherichia coli cells by the antagonism of the B subunit of the bacterial enzyme DNA gyrase. The antagonism may be by the growth of cells (i) at semipermissive temperatures in a bacterial mutant containing a thermolabile gyrase B subunit or (ii) at semipermissive concentrations of coumermycin A1, an antibiotic that specifically inhibits the B subunit of DNA gyrase. The kinetics of plasmid elimination indicate that plasmid loss occurs too rapidly to be explained solely by the faster growth of that plasmid-free bacteria and, therefore, represents interference with plasmid maintenance. PMID- 6288664 TI - Cloning and identification of the product of the dnaE gene of Escherichia coli. AB - We successively subcloned the dnaE gene of Escherichia coli into pBR322, resulting in a plasmid that contains 4.6 kilobases of E. coli DNA. This plasmid can complement a dnaE temperature-sensitive mutation. A restriction map of the dnaE gene and the surrounding 10.7-kilobase region of the E. coli chromosome was determined. A unique HindIII restriction endonuclease site within the cloned segment of DNA was identified as a site required for expression of the dnaE gene. By using the maxicell plasmid-directed protein synthesizing system, we demonstrated that dnaE codes for the alpha subunit of DNA polymerase III. PMID- 6288665 TI - Electron transport components involved in hydrogen oxidation in free-living Rhizobium japonicum. AB - Membranes from free-living Rhizobium japonicum were isolated to study electron transport components involved in H2 oxidation. The H2/O2 uptake rate ratio in membranes was approximately 2. The electron transport inhibitors antimycin A, cyanide, azide, hydroxylamine, and 2-n-heptyl-4-hydroxyquinoline-N-oxide (HQNO) inhibited H2 uptake and H2-dependent O2 uptake significantly. H2-reduced minus O2 oxidized absorption difference spectra revealed peaks at 551.5, 560, and 603 nm, indicating the involvement of cytochromes c, b, and a-a3, respectively. H2 dependent cytochrome reduction was completely inhibited in the presence of 0.15 mM HQNO. This inhibition was relieved by the addition of 0.1 mM menadione. Evidence is presented for the involvement of two b-type cytochromes in H2 oxidation. One b-type cytochrome was not reduced by ascorbate and had an absorption peak at 560 nm. The reduction of this cytochrome by H2 was not inhibited by cyanide. A second b-type cytochrome, cytochrome b', was not reduced by H2 in the presence of cyanide. This cytochrome had an absorption peak at 558 nm. Carbon monoxide difference spectra with H2 as reductant provided evidence for the involvement of cytochrome o as well as cytochrome a3 in H2 oxidation. H2 uptake activity in cell-free extracts was inhibited by UV light irradiation. Most of the activity of the UV-treated extracts was restored with the addition of ubiquinone. The restored activity was inhibited by cyanide. A branched electron transport pathway from H2 to O2 is proposed. PMID- 6288667 TI - Plasmid transfer to Bordetella pertussis: conjugation and transformation. AB - Plasmids of the P and W incompatibility groups were introduced into Bordetella pertussis by conjugation. Plasmid DNA isolated from B. pertussis could be reintroduced by transformation. DNA isolated from Escherichia coli could not be introduced into B. pertussis by transformation if this DNA contained HindIII restriction sites. We have demonstrated that HindIII sites are modified by B. pertussis. Plasmids of the FI and FII incompatibility groups could not be introduced into B. pertussis by conjugation, and nonconjugative plasmids of the ColE1 and Q incompatibility groups could not be introduced by transformation. Our ability to introduce plasmids in the laboratory suggests that the apparent lack of plasmids in natural isolates of B. pertussis is not due to an inability to act as a plasmid recipient. PMID- 6288666 TI - Identification of an insertion sequence in the chromosome of Pseudomonas aeruginosa PAO. AB - An insertion sequence, IS22, in Pseudomonas aeruginosa PAO was identified by its genetic properties. It occurred as a single copy in the chromosome, as shown by Southern hybridization. A restriction map of it was constructed. PMID- 6288668 TI - Study on the interaction between hemoglobin Izu (Macaca) and organic phosphates by spin labeling. AB - ESR spectra of the carbonmonoxy, oxy, and deoxy derivatives of hemoglobin Izu [Hb Izu (Macaca): beta 83 (EF 7) Gly leads to Cys] labeled at cysteine beta 83 with maleimide spin label have been observed in the presence and absence of 2,3 diphosphoglycerate and inositol hexaphosphate. The tau c values obtained from the spectra indicated that inositol hexaphosphate binds to all the derivatives of Hb Izu, but 2,3-diphosphoglycerate only to the deoxy derivatives. PMID- 6288669 TI - Effect of polyamines on two types of reaction of purified poly(ADP-ribose) polymerase. AB - The effects of polyamines on reactions catalyzed by bovine thymus poly(ADP ribose) polymerase were examined under various conditions, and the following results were obtained. (1) Spermine and spermidine, and putrescine to a lesser degree can stimulate the synthesis of poly(ADP-ribose) covalently bound to the enzyme without Mg2+ and histones. (2) A part of the above stimulation can be explained by the Mg2+-sparing effect of polyamines. (3) The other part of the stimulation is shown to be through protection of the enzyme against the formation of an abortive complex of the enzyme and denatured DNA, which contaminates some native DNA preparations used for enzyme activation. Similar protection was shown earlier in this laboratory with histones. (4) Putrescine seems to lack this enzyme-protecting activity. (5) The polyamine effect observed in the Mg2+ dependent reaction is variable depending on the DNA preparations used. (6) Chemical analysis shows that the average chain lengths of the products synthesized with Mg2+ and spermine are similar, and the products are covalently bound to the enzyme, indicating that the reaction supported by polyamines is essentially the same as that by Mg2+. (7) Under the histone H1-dependent reaction conditions where ADP-ribosylation of histone H1 is predominant, both Mg2+ and polyamines are inhibitory on the reaction and both cations decrease the number of product molecules without affecting the size of the product. These data suggest that polyamines can at least partially replace Mg2+ in terms of effect on the ADP ribosylation reaction. The other effect of polyamines is the protection of the enzyme from abortive binding to denatured DNA, as has also been shown to occur with histones. PMID- 6288670 TI - Incorporation of a ganglioside and spin-labeled ganglioside analogue into cell and liposomal membranes. AB - When an aqueous solution of a spin-labeled "two tail" gangliosidoid was incubated with liposomes or sheep erythrocytes, the broad single resonance line in the ESR spectrum disappeared and a signal showing an anisotropic motion appeared, indicating that the spin-labeled "two tail" gangliosidoid in the micellar state was transferred to the lipid phase of the acceptor membranes. The transfer was temperature- and time-dependent, irrespective of the acceptor membranes, indicating that the rate of transfer is determined by the escape of monomers from the micelles. The kinetics and temperature-dependence of the association of ganglioside II3NeuAc-GgOse4Cer with sheep erythrocytes was very similar to that of the "two tail" gangliosidoid, indicating that parts of ganglioside II3NeuAc GgOse4Cer could be incorporated into the lipid phase of membranes via a similar mechanism. PMID- 6288671 TI - Interaction of sodium and potassium ions with Na+,K+-ATPase. I. Ouabain-sensitive alternative binding of three Na+ or two K+ to the enzyme. PMID- 6288672 TI - Interaction of sodium and potassium ions with Na+,K+-ATPase. II. General properties of ouabain-sensitive K+ binding. AB - General properties of ouabain-sensitive K+ binding to purified Na+,K+-ATPase [EC 3.6.1.3] were studied by a centrifugation method with 42K+. 1) The affinity for K+ was constant at pH values higher than 6.4, and decreased at pH values lower than 6.4. 2) Mg2+ competitively inhibited the K+ binding. The dissociation constant (Kd) for Mg2+ of the enzyme was estimated to be about 1 mM, and the ratio of Kd for Mg2+ to Kd for K+ was 120 : 1. The order of inhibitory efficiency of divalent cations toward the K+ binding was Ba2+ congruent to Ca2+ greater than Zn2+ congruent to Mn2+ greater than Sr2+ greater than Co2+ greater than Ni2+ greater than Mg2+. 3) The order of displacement efficiency of monovalent cations toward the K+ binding in the presence or absence of Mg2+ was Tl+ greater than Rb+ greater than or equal to (K+) greater than NH4+ greater than or equal to Cs+ greater than Na+ greater than Li+. The inhibition patterns of Na+ and Li+ were different from those of other monovalent cations, which competitively inhibited the K+ binding. 4) The K+ binding was not influenced by different anions, such as Cl-, SO4(2-), NO3-, acetate, and glycylglycine, which were used for preparing imidazole buffers. 5) Gramicidin D and valinomycin did not affect the K+ binding, though the former (10 micrograms/ml) inhibited the Na+,K+-ATPase activity by about half. Among various inhibitors of the ATPase, 0.1 mM p chloromercuribenzoate and 0.1 mM tri-n-butyltin chloride completely inhibited the K+ binding. Oligomycin (10 micrograms/ml) and 10 mM N-ethylmaleimide had no effect on the K+ binding. In the presence of Na+, however, oligomycin decreased the K+ binding by increasing the inhibitory effect of Na+, whether Mg2+ was present or not. 6) ATP, adenylylimido diphosphate and ADP each at 0.2 mM decreased the K+ binding to about one-fourth of the original level at 10 microM K+ without MgCl2 and at 60 microM K+ with 5 mM MgCl2. On the other hand, AMP, Pi, and p-nitrophenylphosphate each at 0.2 mM had little effect on the K+ binding. PMID- 6288673 TI - Re-examination of the ColE1 DNA regions essential for autonomous replication and their functions. AB - Starting from pAO3, a plasmid consisting of a quarter of colicinogenic factor E1 (ColE1) DNA, various small ColE1 derivatives were constructed by in vitro recombination and their ability to achieve autonomous replication was examined. The 436 base pair HaeIII-C fragment of pAO3 contained information for replication when it was recombined with the non-replicating Amp fragment. However, when it was connected to other DNA fragments, the resulting hybrid molecules were not isolated as plasmids. The present results indicate that the additional region of about 240 base pairs next to the HaeIII-C fragment of ColE1 is also essential for the maintenance of a plasmid state. Moreover, using various small ColE1 derivatives, the DNA region responsible for the interference and incompatibility functions of ColE1 DNAs was located. The results indicate that the interference and incompatibility functions are coded by the same ColE1 DNA segment and are not essential for the maintenance of a plasmid state. PMID- 6288674 TI - The importance of membrane integrity in kinetic characterizations of the microsomal glucose-6-phosphatase system. AB - The transport model of glucose-6-phosphatase (EC 3.1.3.9) was recently challenged by a report that detergent treatment had no effect on the presteady state kinetics of glucose-6-P hydrolysis catalyzed at 0 degree C by the enzyme in liver microsomes previously frozen in 0.25 M mannitol (Zakim, D., and Edmondson, D. E. (1982) J. Biol. Chem. 257, 1145-1148). The lack of response to detergent is shown to be the expected consequence of the conditions used in the presteady state measurements. First, when the assay temperature was reduced from 30 to 0 degree C the depression in the glucose-6-P phosphohydrolase activity of intact microsomes (i.e. the system) was much greater than that of fully disrupted microsomes (i.e. enzyme). This indicates that temperature influences transport much more than hydrolysis of glucose-6-P. As a result, the contribution of a small fraction of enzyme associated with disrupted structures is markedly exaggerated, so it becomes the predominant hydrolytic activity before detergent treatment. Second, freezing microsomes in 0.25 M mannitol caused such extensive disruption that all of the activity manifest at 0 degree C could be attributed to enzyme in disrupted structures. The present findings underscore the importance of assessing the state of intactness of "untreated" microsomes and quantifying the contribution of the disrupted component in kinetic analyses of the glucose-6-phosphatase system. The proposition that the detergent-induced changes in the kinetic properties of glucose 6-phosphatase represent removal of constraints imposed on the enzyme by the membrane environment rather than increased access of enzyme to substrate is critically analyzed. PMID- 6288675 TI - Regulation of ovarian progestin production by epidermal growth factor in cultured rat granulosa cells. AB - The modulation of ovarian steroidogenesis by epidermal growth factor (EGF) was investigated in cultured rat granulosa cells. Granulosa cells, obtained from ovaries of immature, hypophysectomized, estrogen-treated rats, were incubated for 2 days with EGF, follicle-stimulating hormone (FSH), or EGF plus FSH. Treatment with EGF did not affect estrogen production, but stimulated progestin (i.e. progesterone and 20 alpha-hydroxy-pregn-4-en-3-one) production in a dose dependent manner. Stimulation of progestin production by EGF appears to be the result of an increase in pregnenolone biosynthesis as well as increases in the activities of 20 alpha-hydroxysteroid dehydrogenase and 3 beta-hydroxysteroid dehydrogenase/isomerase. Treatment with FSH increased both estrogen and progestin production by cultured granulosa cells. When cells were treated concomitantly with EGF, FSH-stimulated estrogen production was inhibited, while progestin production was further enhanced. The EGF enhancement of FSH-stimulated progestin production appears to be the result of synergistic increases in pregnenolone biosynthesis and 20 alpha-hydroxysteroid dehydrogenase activity, resulting in substantial increases in 20 alpha-hydroxypregn-4-en-3-one but not progesterone production. The effects of EGF were shown to be time-dependent. The concept of a direct action of EGF on rat granulosa cells is reinforced by the demonstration of high affinity (Kd approximately 3 X 10(-10) M), low capacity (approximately 5,000 sites/cell) EGF binding sites in these cells. Thus, EGF interacts with specific granulosa cell receptors to stimulate progestin but to inhibit estrogen biosynthesis. PMID- 6288676 TI - The catalytic mechanism of cytochrome P-450. Spin-trapping evidence for one electron substrate oxidation. AB - Cytochrome P-450 is destroyed during catalytic oxidation of several 4-substituted 3,5-bis(ethoxycarbonyl)-2,6-dimethyl-1,4-dihydropyridine substrates. A qualitative correlation has been found between the ability to destroy cytochrome P-450 and the stability of the 4-substituent as a radical. Destruction of the enzyme by the 4-ethyl (DDEP), 4-propyl, and 4-isobutyl analogues is due to transfer of the 4-alkyl group from the substrate to a nitrogen of the prosthetic heme, a process which gives rise to isolable N-alkylprotoporphyrin IX derivatives. Little enzyme destruction is observed when the 4-alkyl group is of low radical stability (methyl, phenyl) and good destruction, but no isolable heme adducts when the 4-substituent is of very high radical stability (isopropyl, benzyl). Spin-trapping studies have established that the 4-ethyl group in DDEP is lost as a radical as a result of oxidation by cytochrome P-450. Of three commonly used spin traps, only alpha-(4-pyridyl-1-oxide) N-tert-butylnitrone was found suitable for such studies. The other spin traps, 5,5-dimethyl-1-pyrroline-N-oxide and alpha-phenyl N-tert-butylnitrone, were found to be ineffective, the latter because it strongly inhibits cytochrome P-450. Hydrogen peroxide formed in situ can support a part of the cytochrome P-450-catalyzed ethyl radical formation and DDEP-dependent self-inactivation. The results provide persuasive evidence that oxidation of the nitrogen in DDEP by cytochrome P-450 proceeds in one-electron steps. Cytochrome P-450 may thus function, at least with certain substrates, as a one-electron oxidant. PMID- 6288677 TI - Rat liver microsomal glucose-6-P translocase. Effect of physiological status on inhibition and labeling by stilbene disulfonic acid derivatives. AB - Intact microsomes from groups of fed, fasted, glucocorticoid-treated (triamcinolone) and diabetic (alloxan) rats were reacted with 4,4' diisothiocyanostilbene-2,2'-disulfonic (DIDS), a specific inhibitor of microsomal glucose-6-P translocase. The concentrations that inhibit by 50% were 41 +/- 2, 31 +/- 1, 39 +/- 4, and 18 +/- 1 microM (mean +/- S.E.; n = 3); (order as above). The maximal levels of inhibition of the translocase by DIDS were 66 +/- 2, 79 +/- 2, 63 +/- 1, and 88 +/- 1%, respectively. The differences in the values for the different groups of animals are statistically significant, except for comparisons between fed and triamcinolone-treated animals. Microsomes from the same groups of animals were treated with the tritiated reduced derivative of DIDS, [3H]H2DIDS, which labels a 54,000-dalton polypeptide, previously implicated as a component of the glucose-6-P translocase. The mean values (+/- S.E.) of [3H]H2DIDS bound to the polypeptide under saturating conditions were 100 +/- 6, 120 +/- 9, 62 +/- 7, and 101 +/- 15 pmol/mg of microsomal protein, respectively. The amount bound in microsomes from triamcinolone-treated rats is significantly lower from the values for the other three physiological states, which do not differ significantly from each other. The presence of glucose-6-P, but not mannose-6-P, during the [3H]H2DIDS reaction significantly stimulates the labeling of the 54,000-dalton polypeptide in microsomes from all the classes of animals above, except the diabetic animals. These results indicate that DIDS and [3H]H2DIDS are probes sensitive enough to discern differences in the translocase due to physiological regulation. On the basis of the labeling studies with [3H]H2DIDS, the increase in translocase activity observed in microsomes from fasted, triamcinolone-treated, and diabetic rats cannot be ascribed to increased numbers of translocase molecules, but rather to increased functional activity of the translocase protein. PMID- 6288678 TI - Specific binding of 125I-human interferon-gamma to high affinity receptors on human fibroblasts. AB - Highly purified human interferon-gamma (IFN-gamma) was iodinated with 125I-Bolton Hunter reagent to a specific activity of 34 microCi/micrograms of protein. After iodination, molecular sieve chromatography was used to isolate fractions containing IFN-gamma at approximately 80-90% radioactive purity. This preparation of 125I-IFN-gamma bound specifically to high affinity binding sites on human GM 258 fibroblasts. Scatchard analysis of binding data revealed the presence of 2400 binding sites/cell, each binding with an apparent Kd of 1.5 X 10(-10) M. Binding was competitively inhibited in the presence of unlabeled IFN-gamma and, to a lesser degree, by IFN-beta, but not by IFN-alpha. Neutralizing antibodies specific for IFN-gamma efficiently inhibited the specific binding of 125I-IFN gamma to cells. We conclude that the specific high affinity binding site is the receptor for IFN-gamma. PMID- 6288680 TI - Independent locations of kinase and 3'-phosphatase activities on T4 polynucleotide kinase. AB - We have used two chemical modification reagents and three proteases to study the relationship between the two activities of T4 polynucleotide kinase. In each case, conditions were found where one of the two activities of the enzyme could be eliminated without greatly reducing the other. Taken together, these data indicate that the two activities are catalyzed by amino acid residues located in separate active sites on the polypeptide chain. Specific exopeptidase digestion indicates that the kinase activity lies in the NH2-terminal and the phosphatase in the COOH-terminal portion of the polypeptide chain. Partial trypsin digestion produces a 29,000-dalton fragment with no kinase activity and nearly normal 3' phosphatase activity. PMID- 6288679 TI - Isolation and characterization of two mutant forms of T4 polynucleotide kinase. AB - The purification of polynucleotide kinase from Escherichia coli infected by two different mutants in the T4 polynucleotide kinase (pseT) gene is described. The pseT 1 enzyme has virtually no 3' specific phosphatase activity and normal polynucleotide kinase activity. The pseT 47 enzyme has very little phosphatase activity and no kinase activity. However, enzyme isolated from a pseT 1, pseT 47 mixed infection appears to contain heterodimers with considerably more phosphatase activity. Thus, the pseT 47 mutation partially inactivates the phosphatase and totally inactivates the kinase. A study of the action of polynucleotide kinase on plasmid DNAs nicked to give a 3'-phosphate and a 5' hydroxyl indicates that although the enzyme can catalyze both the removal of the 3'-phosphate and the insertion of a 5'-phosphate, there is no evidence for a concerted reaction involving both activities on the same polypeptide chain. PMID- 6288681 TI - Isolation and characterization of a 94,000-dalton protein with thyrotropic activity from early bovine placenta. AB - A thyrotropic protein was extracted and purified from the placenta of early bovine gestations. After protein extraction, the 45-60% ammonium sulfate precipitate of maternal and fetal bovine cotyledons was found to compete with thyroid stimulating hormone (TSH) for binding to thyroid cell membranes and to mediate TSH specific biological effects including the stimulation of cyclic AMP production, iodide uptake, and thyroxine secretion. The placental thyrotropin was further purified by gel and anion exchange chromatography, followed by binding to thyroid cell membranes and elution by mild acid treatment. 400 micrograms of isolated protein with 4.5 units of TSH-like binding activity/mg of protein was recovered from the placenta of a 90-day-old bovine gestation, representing 2 X 10(-4%) of its original wet weight. The placental thyrotropin appeared to be a 94,000-dalton protein with pI 6.0 and composed of two noncovalently associated chains of 50,000 and 44,000 daltons. The placental 94,000-dalton thyrotropin bound to TSH membrane receptors and induced specific TSH-mediated biological effects, but was structurally and immunologically distinct from TSH and hypophysical or placental gonadotropins. PMID- 6288682 TI - Epstein-Barr virus induction by a serum factor. Purification of a high molecular weight protein that is responsible for induction. AB - Serum contains a factor that induces Epstein-Barr virus antigens in latently infected human lymphoblastoid cell lines and that cooperates with chemical inducers. Here we report the purification of a protein that is responsible for the effect. Using ammonium sulphate precipitation, chromatography on DEAE-agarose and cellulose, molecular sieve chromatography on Bio-Gel A-5, and glycerol gradient centrifugation, the factor was enriched about 300,000-fold compared to calf serum. Under neutral conditions, the protein chromatographed as a high molecular weight protein of about 5.5 X 10(5) both in its active and inactive form. Both forms of the molecule sedimented at about 7 s. The factor is an acidic protein with a pI of 5. It seems to be composed of high molecular and low molecular weight subunits. Nanogram amounts of purified factor are sufficient for measurable inducing effects. PMID- 6288683 TI - Epstein-Barr virus induction by a serum factor. Characterization of the purified factor and the mechanism of its activation. AB - Purified Epstein-Barr virus-inducing factor shows all the properties previously described for crude serum, i.e. inducing activity, cooperative activity with the inducers 12-O-tetradecanoyl-phorbol-13-acetate, 5'-iodo-2'-deoxyuridine, N butyric acid, anti-IgM, and the need to be activated by pH shock. A small subpopulation of untreated factor molecules is fully active, however. The factor is a protein that is characterized by considerable heat resistance and sensitivity to reducing agents. It does not seem to require sugar residues or lipid for its activity. Activation occurs both at alkaline and acidic pH. Inactive and activated factor differ in their sensitivity to trypsin and in the degree of sensitivity to reducing agents. A change of conformation within the molecule seems to be the biochemical basis for the activation reaction. PMID- 6288684 TI - The guanine nucleotide activating site of the regulatory component of adenylate cyclase. Identification by ligand binding. PMID- 6288685 TI - The involvement of hydroxyl radical derived from hydrogen peroxide in lignin degradation by the white rot fungus Phanerochaete chrysosporium. AB - The possible involvement of hydrogen peroxide (H2O2)-derived hydroxyl radical (.OH) in lignin degradation ([14C]lignin leads to 14CO2) by Phanerochaete chrysosporium was investigated. When P. chrysosporium was grown in low nitrogen medium (2.4 mM N), an increase in the specific activity for H2O2 production in cell extracts was observed to coincide with the appearance of ligninolytic activity and both activities appeared after the culture entered stationary phase. The production of .OH in ligninolytic cultures of P. chrysosporium was demonstrated by alpha-keto-gamma-methiolbutyric acid-dependent formation of ethylene. Hydrogen peroxide-dependent .OH formation was also shown in cell extracts of ligninolytic cultures. The radical species was demonstrated to be .OH by the .OH-dependent hydroxylation of p-hydroxybenzoic acid to form protocatechuic acid and by using 5,5-dimethyl-1-pyrroline-N-oxide and detecting the production of the nitroxide radical of 5,5-dimethyl-1-pyrroline-N-oxide by EPR. These reactions were inhibited by .OH-scavenging agents and were stimulated when azide was added to inhibit endogenous catalase. Lignin degradation by P. chrysosporium was markedly suppressed in the presence of the .OH-scavenging agents mannitol, benzoate, and the nonspecific radical scavenging agent butylated hydroxytoluene. The above results indicate that .OH derived from H2O2 is involved in lignin biodegradation by P. chrysosporium. PMID- 6288686 TI - Synthesis, turnover, and down-regulation of epidermal growth factor receptors in human A431 epidermoid carcinoma cells and skin fibroblasts. AB - Epidermal growth factor (EGF) receptors extracted with Triton X-100 from human skin fibroblasts and A431 epidermoid carcinoma cells rapidly lose EGF-binding activity precipitable with polyethylene glycol. The presence of concanavalin A which can cross-link and, thereby, aggregate the receptors, allowed quantitative recovery of the lost EGF-binding activity. Scatchard analysis of EGF binding of Triton X-100-solubilized receptors showed that A431 cells and skin fibroblasts possess approximately 1.5 X 10(6) and 7 X 10(4) EGF-binding sites/cell, respectively, which exhibit similar affinities for the ligand. The heavy isotope density-shift method was employed to determine whether differences in rates of receptor synthesis or decay account for the large difference in number of receptors/cell between the two cell types. After shifting cells to medium containing heavy (15N, 13C, and 2H) amino acids, light and heavy receptors, solubilized from total cellular membranes, were resolved by isopycnic banding on density gradients and then quantitated. It was demonstrated that A431 cells synthesize EGF receptors at a rate 12 times faster than skin fibroblasts and that the half-life for receptor decay of A431 cells is somewhat longer (t1/2 = 16 h) than that (t1/2 = 9 h) of fibroblasts. Down-regulation of cell surface and total cellular EGF-binding capacity in A431 cells occurs with a t1/2 of 2-3 h and results in a 70-83% decrease in receptor level in 12 h. Scatchard analysis revealed that these changes in EGF binding were due to an alteration of receptor number and not EGF-binding affinity. Rates of EGF receptor synthesis and inactivation/decay were determined by the heavy isotope density-shift method. No change in the rate of receptor synthesis occurred as a consequence of EGF receptor down-regulation. Down-regulation, however, caused a decrease in receptor half-life from 16 to 4.5 h. These results indicate that EGF-dependent regulation of EGF receptor level in A431 cells involves an alteration of the rate of receptor inactivation. PMID- 6288687 TI - Specific molecular activities of recombinant and hybrid leukocyte interferons. AB - Hybrid interferon DNA recombinants were constructed from the IFLrA and IFLrD leukocyte interferon-coding sequences. Each of the hybrid interferons was purified with the use of a monoclonal antibody to human leukocyte interferon. Three amino acid residues were identified, one or all of which function to potentiate antiviral activity on feline cells and reduce activity on human cells. Because at sufficiently high concentrations human interferons can interact with mouse and rat receptors, it is apparent that the species barrier is only relative and that interferons can be forced into heterologous receptors by mass action. In addition, the specific molecular antiviral and antiproliferative activities (molecules of interferon/cell required for a specific effect) for each of these interferons were determined. The specific molecular activities permit an accurate comparison of the efficacy of different interferons for a specific effect. Because the ratios of antiproliferative/antiviral activity of these interferons vary over a 12-fold range, it appears that the antiviral and antiproliferative activities are promulgated through different mechanisms. To account for these results, it is proposed that there are at least two distinct interferon receptors on cells. PMID- 6288688 TI - Differential effects of single and repeated administration of gonadotropins on testosterone production and steroidogenic enzymes in Leydig cell populations. PMID- 6288689 TI - The reactivity of Mg-substituted horseradish peroxidases. AB - Mg-substituted horseradish peroxidases were oxidized by K2IrCl6 or K3Fe(CN)6 to their porphyrin radical form and the 1:1 stoichiometric relationship was confirmed by spectrophotometric, fluorophotometric, and ESR titration methods. The values of E'0 for oxidations of Mg peroxidases A and C were both 0.63 V at pH 6 and depended on pH in the same way as postulated for the Compound I/Compound II couples of the corresponding enzymes. Unlike Zn peroxidase C, Mg peroxidase C was not directly oxidized by H2O2. The oxidation was catalyzed by the native peroxidase. Mg peroxidase C was photooxidized to the radical form faster than Zn peroxidase C, but its oxidation was accomplished by irreversible changes in the porphyrin in the early stage of reaction. The rate of reduction of the oxidized Mg peroxidases in the presence of various electron donors was measured at varying pH values and compared with the rate of Compound I reduction. A role of porphyrin as a site of electron transfer in the peroxidase catalysis was suggested. PMID- 6288690 TI - The gelatinolytic activity of human skin fibroblast collagenase. AB - The gelatinolytic activity of human skin fibroblast collagenase was examined on denatured collagen types I-V. All denatured substrates were cleaved, including types IV and V, which are resistant to collagenase in native form. Interestingly, the earliest major cleavage in denatured collagen types I-III occurred at a 3/4 1/4 locus, resulting in products electrophoretically identical with TCA and TCB fragments of mammalian collagenase action on these native collagens. However, in the denatured substrates, multiple additional proteolytic cleavages followed. The propensity for cleavage at a 3/4-1/4 site in denatured collagen, where sequence is the major specifier of enzymatic action, would seem to indicate that the most favorable amino acid sequence of gamma chains for catalysis is located in this region. The peptide bond specificity of human fibroblast collagenase on gelatin was examined by amino acid sequencing of extensively cleaved denatured type I collagen. Analysis of the NH2-terminal amino acid residues from the resultant gelatin peptides showed sequences of "-H2N-Ile-Y-Gly" and "H2N-Leu-Y-Gly" only (where Y indicates that any amino acid can be found in that position), indicating that Gly-Ile and Gly-Leu bonds are the only sites of collagenase cleavage in this substrate. Whereas the gamma1 chains of denatured collagen types I-III were cleaved at similar rates, fibroblast collagenase was a much better gamma2 gelatinase than gamm1-gelatinase on denatured type 1 collagen. This preference for the cleavage of gamma2(I) was the result of both a higher kcat (750 versus 230 h-1) and lower Km (3.7 versus 7.0 microM) than for a gamma1(1), resulting in an overall selectivity (kcat/Km) of greater than 6-fold. Compared to such kinetic parameters on native collagen, these values indicate that gelatinolysis is somewhat slower than collagenolysis. PMID- 6288691 TI - Follitropin binding to receptors in testis. Modulation by monovalent salts and divalent cations. PMID- 6288692 TI - Hepatic alpha 1-adrenergic receptors show agonist-specific regulation by guanine nucleotides. Loss of nucleotide effect after adrenalectomy. PMID- 6288693 TI - Guanosine 5'-(beta, gamma-imido)triphosphate inhibition of forskolin-activated adenylate cyclase is mediated by the putative inhibitory guanine nucleotide regulatory protein. PMID- 6288694 TI - Cyclic nucleotide-dependent protein kinases in airway smooth muscle. AB - Because of the potential importance of cyclic nucleotide-dependent protein kinases in the regulation of airway smooth muscle tone, we have examined some of the characteristics of these enzymes in the soluble fraction of canine trachealis homogenates. In the absence of added cAMP, the heat-stable cAMP-dependent protein kinase inhibitor (PKI) abolished only a half of the 32P incorporation into mixed histones. The remaining activity appeared to be contributed by a cyclic nucleotide-independent enzyme. Phosphotransferase activity was enhanced 5-fold by 5 microM cAMP but only 70% of the cAMP-stimulated activity could be inhibited by PKI. The sensitivity of the cyclic nucleotide-dependent, PKI-resistant enzyme to cAMP, cGMP, and Mg2+ indicated that it was cGMP-dependent protein kinase. Because of the large amount of cyclic nucleotide-independent activity, and the ability of cAMP to activate cGMP-dependent protein kinase, the traditional "-cAMP/+cAMP" ratio did not provide an accurate assessment of the in vivo activation state of cAMP-dependent protein kinase. However, a modified assay was developed which allowed the precise measurement of cAMP-dependent, cGMP-dependent, and cyclic nucleotide-independent protein kinase activities. Using this new method, the cAMP dependent protein kinase activity ratio of 0.239 in untreated trachealis strips was increased to 0.355 and 0.386 by prior exposure of the intact tissue to the smooth muscle relaxants isoproterenol and prostaglandin E2, respectively. The results of this study are consistent with the proposed role of cAMP-dependent protein kinase in the regulation of smooth muscle contractile function. PMID- 6288695 TI - Escherichia coli glutaminyl-tRNA synthetase. I. Isolation and DNA sequence of the glnS gene. AB - We have isolated a lambda-transducing phage carrying the gene (glnS) for Escherichia coli glutaminyl-tRNA synthetase. The location of the glnS gene within the 13.5-kilobase E. coli DNA transducing fragment was determined by genetic means. The glnS gene was recloned into plasmid pBR322 and its nucleotide sequence was established. The DNA sequence translates to a protein of 550 amino acids. PMID- 6288696 TI - A termination site for LacI transcription is between the CAP site and the lac promoter. PMID- 6288697 TI - Labeling of succinate-cytochrome c reductase with 125I. Accessibility of the peptides to the aqueous phases on the cytosolic and matrix sides of the mitochondrial membrane. AB - Lactoperoxidase-catalyzed radioiodination was used to study the arrangement of the component peptides of succinate-cytochrome c reductase with respect to the aqueous phases on each side of the mitochondrial inner membrane. Mitochondria depleted of their outer membrane and inside-out vesicles purified from submitochondrial particles by the lectin-affinity procedure (D'Souza, M. P., and Lindsay, J. G. (1981) Biochim. Biophys. Acta 640, 463-472) were iodinated using immobilized preparations of lactoperoxidase. The labeled membranes were solubilized in detergent and the succinate-cytochrome c reductase was purified by immunoprecipitation with specific IgG. Analysis of the radioiodine distribution after sodium dodecyl sulfate-polyacrylamide gel electrophoresis and comparison with peptide stain patterns show that bands 2 (64 kilodaltons), 6 (30 kilodaltons), 9 (15 kilodaltons), and 11 (less than 10 kilodaltons) are labeled from the cytoplasmic surface of the membrane. Bands 1 (72 kilodaltons), 4 (48 kilodaltons), and 8 (20 kilodaltons) appear to be labeled on the matrix side of the membrane, while bands 3 (52 kilodaltons), 5 (35 kilodaltons), 7 (25 kilodaltons), and 10 (11 kilodaltons) are labeled from both sides of the membrane. Tentative identification of the labeled bands suggests that band 1 is the large subunit of succinate dehydrogenase. Bands 3 and 4 represent proteins which have been referred to as core proteins I and II. Bands 5 and 6 are the proteins associated with cytochromes b and c1, respectively; band 7 is the Rieske iron-sulfur protein. PMID- 6288698 TI - Characterization of the phosphorylation sites and the surrounding amino acid sequences of the p21 transforming proteins coded for by the Harvey and Kirsten strains of murine sarcoma viruses. AB - The transforming protein coded for by the onc gene (v-rasHa) of Harvey murine sarcoma virus (Ha-MuSV) is the 21,000-dalton protein (p21) which is the immediate agent responsible for the virus-induced malignant transformation of normal cells. The p21 proteins of Ha-MuSV and the closely related Kirsten murine sarcoma virus are heavily phosphorylated in vivo. In the partially purified Ha-MuSV p21, the protein shows a guanine nucleotide-binding activity and, in addition, a very unique autophosphorylating activity at a threonine residue using as phosphoryl donor GTP but not ATP. In the present study, we compared the tryptic peptide maps of the Ha-MuSV p21 phosphorylated in vivo and in vitro. The results show that the major phosphorylation site is identical. Since the GTP-specific phosphorylation is very unique and distinct from all other known protein kinases, the present observation suggests that the in vitro enzymatic activity is responsible for the p21 phosphorylation in vivo. We have analyzed the amino acid sequence surrounding the major phosphorylation site of the Ha-MuSV p21 by automated Edman degradations of the tryptic phosphopeptides. Threonine residue 59 from the initiator methionine residue 1 of the p21 protein is the phosphorylated amino acid residue, and the surrounding amino acid sequence is NH2...-Thr-Cys-Leu-Leu-Asp-Ile-Leu-Asp Thr-Thr(P)-Gly-Gln-Glu-Glu-Tyr-...COOH. The p21 proteins of both the Ha-MuSV and the closely related Kirsten murine sarcoma virus share the same phosphopeptide. The amino acid sequence of the phosphorylation site is distinct from all other known protein kinases. PMID- 6288699 TI - Chromatin fragments containing bovine 1.715 g ml-1 satellite DNA. Purification by chromatography on malachite green DNA affinity resin. AB - Chromatin fragments containing bovine 1.715 g ml-1 satellite DNA (1.715 satellite chromatin) were purified in order to test for differences between satellite chromatin and unfractionated chromatin. The final purification procedure involved digestion of isolated nuclei with Eco RI restriction endonuclease, solubilization of a fraction of the chromatin by lysis of the nuclei at low ionic strength, and chromatography of the soluble chromatin fragments on malachite green DNA affinity resin. Digestion of chromatin within steer kidney and thymus nuclei by Eco RI reached limits at which 22% and 37%, respectively, of the potential sites for the enzyme in the 1.715 satellite chromatin were cleaved. A test of the 1.715 g ml-1 satellite DNA extracted from digested thymus nuclei showed it was not significantly nicked at Eco RI sites. Fractionation of soluble chromatin fragments produced by Eco RI digestion on the basis of size using either sucrose gradient centrifugation or chromatin gel electrophoresis yielded 1.715 satellite chromatin of 60-80% purity. However, chromatography of the soluble chromatin fragments on columns of malachite green resin gave 1.715 satellite chromatin of higher purity and in greater amounts than could be obtained with the sucrose gradients or chromatin gels. Using malachite green resin, hundreds of micrograms of 1;715 satellite chromatin could be obtained with a purity that was usually at least 95% as determined by melting of the extracted DNA. PMID- 6288701 TI - The binding of fibrinogen to its platelet receptor. AB - Specific receptors for fibrinogen can be induced on platelets by a variety of stimuli. In this study, two independent approaches have implicated the D domain of fibrinogen in its interaction with the platelet. Immunochemically purified Fab fragments of anti-fibrinogen and anti-D inhibited 125I-fibrinogen binding to platelets in a dose-dependent fashion and platelet aggregation. In contrast, Fab fragments of anti-E produced only a slight inhibition of fibrinogen binding and nonimmune Fab fragments had no effect. Fibrinogen was digested with plasmin in the presence of 5 mM calcium and fragment D and E of Mr 100,000 and 50,000, respectively, were isolated. This D fragment inhibited 125I-fibrinogen binding to platelets in a concentration-dependent fashion, whereas the E fragment was ineffective. With 125I-fibrinogen at 0.17 microM, nonlabeled fibrinogen inhibited binding by 50% at 0.7 microM, whereas 170 microM fragment D was required to produce 50% inhibition. D fragment of Mr 80,000, generated in the absence of calcium, was noninhibitory. These observations provide strong evidence for the participation of the D domain in the binding of fibrinogen by its platelet receptor and suggest that the recognition site is lost in the conversion of the Mr 100,000 to 80,000 D species. PMID- 6288700 TI - Reconstitution of neurotoxin-stimulated sodium transport by the voltage-sensitive sodium channel purified from rat brain. AB - Incorporation of the saxitoxin receptor of the sodium channel solubilized with Triton X-100 and purified 250-fold from rat brain into phosphatidylcholine vesicles is described. Fifty to 80% of the saxitoxin receptor sites are recovered in the reconstituted vesicles (KD = 3 nM). Unlike the detergent-solubilized saxitoxin receptor, the reconstituted saxitoxin binding activity is stable to incubation at 36 degrees C. Approximately 75% of the reconstituted saxitoxin receptor sites are externally oriented and 25% are inside-out. The initial rate of 22Na+ uptake into reconstituted vesicles is increased up to 3- to 4-fold by veratridine with a K0.5 of 11 microM. Seventy per cent of this increase is blocked by external tetrodotoxin (TTX) with a Ki of 10 nM. All of the veratridine stimulated 22Na+ uptake is blocked when TTX is present on both sides of the vesicle membrane, or when tetracaine is added to the external medium. The apparent binding constants for veratridine, saxitoxin, and TTX are essentially identical to those in intact rat brain synaptosomes. The results demonstrate reconstitution of sodium transport, as well as neurotoxin binding and action, from substantially purified sodium channel preparations. PMID- 6288702 TI - Selective inhibition of DNA replication in herpes simplex virus infected cells by 1-(2'-deoxy-2'-fluoro-beta-D-arabinofuranosyl)-5-iodocytosine. PMID- 6288704 TI - Steric inhibition of phenylboronate complex formation of 2'-(5"-phosphoribosyl) 5'-AMP. AB - Unlike all other adenine nucleotides which contain cis-diols in their ribose groups, the degradation product of polyadenosine diphosphoribose, 2'-(5" phosphoribosyl)-5'-AMP, does not bind to boronate affinity columns. This anomalous behavior occurs even under conditions of high ionic strength (1 M salt), indicating that the absence of binding is not due to generalized repulsion between the negatively charged phosphate groups of 2'-(5"-phosphoribosyl)-5'-AMP and the negatively charged boronate sites of the resin. Despite the lack of boronate binding the presence of a chemically functional cis-diol group in 2'-(5" phosphoribosyl)-5'-AMP was verified by quantitative periodate oxidation. Subsequent dephosphorylation rendered the cis-diol in ribosyladenosine accessible to binding to the boronate sites; therefore, one or both of the phosphate groups sterically interfere with complexation of the cis-diol with boronate. Computer model-building studies based on NMR data implicate the 5'-phosphate group of the molecule, is located sterically near the cis-diol group, inhibiting the complex formation with boronate. PMID- 6288703 TI - Stimulation of forskolin of intact S49 lymphoma cells involves the nucleotide regulatory protein of adenylate cyclase. PMID- 6288706 TI - Formation of the semiquinone form in the anaerobic reduction of adrenodoxin reductase by NADPH. Resonance Raman, EPR, and optical spectroscopic evidence. PMID- 6288705 TI - Quantitation and characterization of the (Na+,K+)-adenosine triphosphatase in the rat adipocyte plasma membrane. PMID- 6288707 TI - The nature of CuA in cytochrome c oxidase. AB - The isolation and purification of yeast cytochrome c oxidase is described. Characterization of the purified protein indicates that it is spectroscopically identical with cytochrome c oxidase isolated from beef heart. Preparations of isotopically substituted yeast cytochrome c oxidase are obtained incorporating [1,3-15N2]histidine or [beta,beta-2H2]cysteine. Electron paramagnetic resonance and electron nuclear double resonance spectra of the isotopically substituted proteins identify unambiguously at least 1 cysteine and 1 histidine as ligands to CuA and suggest that substantial spin density is delocalized onto a cysteine sulfur in the oxidized protein to render the site Cu(I)--S. PMID- 6288708 TI - Nucleoside phosphorothioates as probes of the nucleotide binding site of brain pyridoxal kinase. AB - N-Dansyl-2-oxopyrrolidine, a competitive inhibitor with respect to ATP, was used as a probe of the nucleotide binding site of pyridoxal kinase. It binds to an hydrophobic region of the catalytic site with a KD = 6 microM. Time emission anisotropy measurements yielded a rotational correlation time of 38 ns for the bound inhibitor. N-Dansyl-2-oxopyrrolidine is immobilized by strong interactions with the nucleotide binding site. Protein fluorescence quenching was used to determine the dissociation constants of the diastereomers of ATP beta S. Both diastereomers bind with a dissociation constant KD = 25 microM. The kinetic parameters Km and Vmax for the pyridoxal kinase reaction were determined for ATP and the diastereomers of ATP beta S in the presence of Mg(II), Co(II), Zn(II), and Cd(II). With Mg(II), pyridoxal kinase exhibits stereoselectivity for the A diastereomer of ATP beta S, Vmax ratio, A/B = 30. In the presence of Cd(II), the stereoselectivity is reversed and the B diastereomer of ATP beta S is the preferred substrate. As the divalent cations were varied in the series Mg(II), Co(II), Zn(II), and Cd(II), the A/B ratio was progressively lowered to the value of 0.2 found for Cd(II). These data indicate that the divalent cations coordinate to the beta-phosphate group of the nucleoside triphosphate substrates. The results obtained with the diastereomers of ATP alpha S suggest that pyridoxal kinase uses the divalent cation, delta,beta,gamma-bidentate nucleotide chelate as substrate. PMID- 6288709 TI - Mechanism of action of cholera toxin on intact cells. Generation of A1 peptide and activation of adenylate cyclase. AB - When intact mouse neuroblastoma NB cells were incubated with choleragen at 4 degrees C, washed, and incubated at 37 degrees C, activation of adenylate cyclase occurred rapidly after a delay of 15 min. The cells were incubated under the same conditions with 125I-labeled toxin, lysed, and solubilized with sodium dodecyl sulfate under mild conditions. Soluble proteins were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis in the absence of dithiol reductants to separate labeled toxin products. Initially, only 0.1 to 0.2% of the cell associated radioactivity migrated on the gels as the A1 peptide of choleragen. After a 15-min delay, the amount of A1 peptide increased rapidly with time and paralleled the activation of adenylate cyclase. Similar results were observed with human skin fibroblasts, Friend erythroleukemic cells, and II3-alpha-N acetylneuraminosyl-gangliotetraosylceramide-treated rat glioma C6 cells. When toxin-treated NB cells were incubated at increasing temperatures, generation of A1 peptide and activation of adenylate cyclase increased in parallel. Both processes were prevented by incubation of cells at 4 or at 37 degrees C in the presence of anticholeragen antibodies. These results indicate that there is delay both in the formation of A1 peptide and in the activation of adenylase cyclase in intact cells. As A1 is believed to be the catalytically active component of choleragen, it is suggested that the lag period may be related in part to the time required to generate A1 peptide from choleragen. PMID- 6288710 TI - Epidermal growth factor and phorbol ester actions on human osteosarcoma cells. Characterization of response and nonresponsive cell lines. PMID- 6288711 TI - Cyclic nucleotide-dependent phosphorylation in Dictyostelium discoideum amoebae. AB - Stimulation of Dictyostelium discoideum amoebae with cAMP was found to induce the specific phosphorylation of a 47,000 molecular weight protein (pP47). This cellular response to cAMP was developmentally regulated. It was first detected in 3 1/2-h starved cells and appeared to persist throughout the aggregation phase of the cells' life cycle. pP47 phosphorylation was specifically induced by cAMP in that amoebae did not respond to stimulation with 5'-AMP, folic acid, Ca2+, and/or the Ca2+ ionophore A23187. cGMP could elicit pP47 phosphorylation but only at high concentrations. Phosphorylation of pP47 in response to cAMP occurred rapidly (within 5 s). The length of time for which it remained phosphorylated depended upon the concentration of the stimulus. With 10(-6) M cAMP, pP47 was phosphorylated for less than 4 min. If amoebae were stimulated with 10(-4) M cAMP, over 30 min were necessary before pP47 was dephosphorylated. Once dephosphorylated, pP47 could again be phosphorylated upon reapplication of the cAMP stimulus. PMID- 6288712 TI - Regulation of protein phosphorylation in hamster insulinoma cells. Identification of Ca2+-regulated cytoskeletal and cAMP-regulated cytosolic phosphoproteins by two-dimensional electrophoresis. PMID- 6288713 TI - Structural characterization of the asymmetric (17 + 13) S species of acetylcholinesterase from Torpedo. II. Component peptides obtained by selective proteolysis and disulfide bond reduction. PMID- 6288715 TI - UDP-N-acetylglucosamine:lysosomal enzyme precursor N-acetylglucosamine-1 phosphotransferase. Partial purification and characterization of the rat liver Golgi enzyme. PMID- 6288714 TI - Characterization of a hydrophobic, dimeric form of acetylcholinesterase from Torpedo. PMID- 6288717 TI - The beta 1-adrenergic receptor of the turkey erythrocyte. Molecular heterogeneity revealed by purification and photoaffinity labeling. AB - The beta 1-adrenergic receptor of turkey erythrocytes has been purified by a combination of affinity and high performance steric exclusion chromatography. These procedures provide preparations with specific activities of greater than 15,000 pmol/mg of protein with an overall recovery of approximately 30% of the receptor activity solubilized from membrane preparations. Sodium dodecyl sulfate polyacrylamide gel electrophoresis of radioiodinated purified receptor reveals two bands of labeled protein with apparent Mr = 40,000 +/- 2,000 and 45,000 +/- 3,000 in a 3-4:1 ratio. These same two peptides can also be labeled specifically and in approximately the same ration in both membranes and purified preparations using the photoaffinity probe 125I-labeled p-azidobenzylcarazolol. When the two purified polypeptides are completely separated by high performance liquid chromatography and subjected to detailed ligand binding studies, identical beta 1 adrenergic specificities are found for the two receptor forms. Preliminary characterization of these two proteins by partial protease digestion suggests a large degree of similarity between them, albeit with some significant differences. These results demonstrate that both purification and photoaffinity labeling identify two polypeptides in turkey erythrocyte membranes as containing a beta 1-adrenergic receptor binding site. The functional and structural relationships of these two forms of the receptor remain to be elucidated. PMID- 6288716 TI - 125I-labeled p-azidobenzylcarazolol, a photoaffinity label for the beta adrenergic receptor. Characterization of the ligand and photoaffinity labeling of beta 1- and beta 2-adrenergic receptors. PMID- 6288718 TI - Purification and properties of a host cell protein required for poliovirus replication in vitro. AB - A host cell protein required for poliovirus RNA-dependent RNA replicase activity in vitro has been purified several thousand-fold from an uninfected HeLa cell postmitochondrial supernatant. A single protein of apparent Mr = approximately 67,000 daltons and pI 6.3 is associated with this "host factor" activity. Poly(U) Sepharose chromatography of the template-dependent replicase isolated from poliovirus-infected cells results in the complete loss of replicase activity if a salt gradient is used to develop the column. Host factor elutes early in the salt gradient and restores replicase activity to protein fractions eluted later in the gradient. The host factor, estimated to be present at 50,000-100,000 copies/cell, interacts physically with replicase. PMID- 6288720 TI - Bacteriophage T4 gene 41 protein, required for the synthesis of RNA primers, is also a DNA helicase. AB - Bacteriophage T4 gene 41 protein is one of the two phage proteins previously shown to be required for the synthesis of the pentaribonucleotide primers which initiate the synthesis of new chains in the T4 DNA replication system. We now show that a DNA helicase activity which can unwind short fragments annealed to complementary single-stranded DNA copurifies with the gene 41 priming protein. T4 gene 41 is essential for both the priming and helicase activities, since both are absent after infection by T4 phage with an amber mutation in gene 41. A complete gene 41 product is also required for two other activities previously found in purified preparations of the priming activity: a single-stranded DNA-dependent GTPase (ATPase) and an activity which stimulates strand displacement synthesis catalyzed by T4 DNA polymerase, the T4 gene 44/62 and 45 polymerase accessory proteins, and the T4 gene 32 helix-destabilizing protein (five-protein reaction). The 41 protein helicase requires a single-stranded DNA region adjoining the duplex region and begins unwinding at the 3' terminus of the fragment. There is a sigmoidal dependence on both nucleotide (rGTP, rATP) and protein concentration for this reaction. 41 Protein helicase activity is stimulated by our purest preparation of the T4 gene 61 priming protein, and by the T4 gene 44/62 and 45 polymerase accessory proteins. The direction of unwinding is consistent with the idea that 41 protein facilitates DNA synthesis on duplex templates by destabilizing the helix as it moves 5' to 3' on the displaced strand. PMID- 6288719 TI - In vitro copying of viral positive strand RNA by poliovirus replicase. Characterization of the reaction and its products. AB - Poliovirus replicase can be isolated in a form which depends on either oligo(U) or on a host cell protein for the initiation of copying of poliovirion (plus strand) RNA. The product of replicase reactions--initiated either with host factor or with oligo(U)--includes full length (35 S) RNA molecules, largely in double-stranded form, which contain the ribonuclease T1-resistant oligonucleotides of the poliovirus minus strand. For the oligo(U)-stimulated reaction, it is shown that the oligo(U) primer is covalently associated with full length product at its 5'-end. For either the host factor- or oligo(U)-dependent reactions, full length molecules appear only after 15 min of synthesis. The fraction of 35 S product is increased by raising the concentration of the limiting nucleoside triphosphate. The reaction is inhibited by as little as 100 mM salt, although it is stimulated by low (20 mM) salt concentrations. Zinc stimulates overall synthesis, but not the rate of appearance of full length molecules; the reaction is inhibited by agents which chelate zinc. Although synthesis of full length products occurs much more slowly than in the infected cell, this soluble system appears to mimic quite faithfully the initial steps of poliovirus replication. PMID- 6288721 TI - Interaction between calf thymus RNA polymerase II and singly nicked Simian virus 40 DNA. PMID- 6288722 TI - Heterogeneity of beta-adrenoreceptor subtypes in the human placenta. PMID- 6288723 TI - Modulation of endogenous noradrenaline release by prejunctional alpha adrenoreceptors: comparison of a cerebral and peripheral artery. AB - 1 Stimulation-evoked fractional release/pulse of noradrenaline (NA) is markedly different in the rabbit basilar artery as compared to the rabbit ear artery. Therefore, using both agonists and antagonists of prejunctional alpha adrenoreceptors, the hypothesis that there are differences in prejunctional mechanisms between these functionally different arteries was tested. 2 Clonidine (10(-6)M) had similar effects on NA release in both vessels, decreasing release at 2 Hz, but increasing release at 8 Hz. However, 10(-5)M clonidine had different effects in the two vessels. 3 alpha-Adrenoreceptor antagonists, phentolamine (10( 6)M), yohimbine (10(-5)M), and phenoxybenzamine (10(-5)M), all increased NA release to a similar extent in both vessels. However, the effect of phenoxybenzamine was considerably greater than that of the other two antagonists, suggesting that it may have additional sites of action. 4 When the effects of modest drug concentrations were considered, no major qualitative or quantitative differences in the effects of agonists and antagonists on NA release from the two vessels were seen. Thus, the difference in fractional NA release between the basilar and ear arteries cannot be accounted for by differences in prejunctional alpha-adrenoreceptor modulation. PMID- 6288724 TI - Non-specific supersensitivity in rat parotid salivary glands following parasympathectomy. AB - 1 The effect of parasympathetic denervation (Px) of the rat parotid salivary glands has been studied. 2 Px produced a significant reduction in wet and dry weight of the parotid gland compared with the intact contralateral control gland. The DNA content per gland was not reduced indicating that cell death had not occurred but that individual cells were smaller. 3 Supersensitivity to both cholinoreceptor and beta-adrenoreceptor stimulation resulted following Px and was observed as a decrease in the threshold dose and an increase in the maximum secretory response. 4 The supersensitivity to beta-adrenoreceptor stimulation was not accompanied by a change in cyclic AMP production but there was an increase in the density of beta-adrenoreceptor binding sites/mg membrane protein. 5 The relationship of this increased density of receptors to the observed supersensitivity is discussed. PMID- 6288725 TI - 3,4-bis(3'-hydroxyphenyl)hexane--a new mammary tumor-inhibiting compound. Studies on the mechanism of action on the DMBA-induced, hormone-dependent mammary tumor of the rat. AB - Biochemical properties, such as the activities of eight carbohydrate-metabolism linked enzymes and four acid hydrolases, and histological characteristics of growing and regressing DMBA-induced mammary tumors of the SD-rat after ovariectomy or treatment of the host with hexesterol, tamoxifen, and 3,4-bis(3' hydroxyphenyl)hexane were determined. Significant differences were found between growing and regressing tumors regardless of the treatment animals had been subjected to. Only few differences in biochemical parameters could be found within the group of tumors regressing due to the applied therapy. The histological signs of regressing tumors were very diverse, but no phenomenon typical of a specific treatment could be found. It cannot be decided whether the partial antiestrogen, 3,4-bis(3'-hydroxyphenyl)hexane unfolds its antimammary tumor activity by means of its estrogenic or its antiestrogenic potency. The hypothesis that estrogens inhibit mammary tumor growth by directing neoplastic cell metabolism toward secretion is not supported by these findings. PMID- 6288726 TI - [Correlation between protein thiols and the doubling time of various rat hepatomas]. AB - The SH content of the soluble proteins (nanomol./mg protein) from five transplantable rat hepatomas and from the DENA-hepatoma were determined with dithionitrobenzoate (Ellman reagent). Both the total number of thiols as well as the number of SH groups that can be blocked by hydroxypentenal (HPE) increase with increasing growth rate of the tumors. In comparison with the protein thiol content of the slowest growing DENA-hepatoma (doubling time 100 days), the total protein thiols of the fastest growing Yoshida hepatoma (doubling time 2,5 days) increase by 100% and the HPE-sensitive protein thiols by 350%. The total protein thiols are significantly correlated with the growth rate (probability of error 5%), the HPE-sensitive thiols are correlated with high significance (probability of error less than 1%). These results are in accordance with the "Molecular Correlation Concept" of G. Weber and might possibly be understood as a consequence of reprogramming of gene expressions. PMID- 6288727 TI - Surface distribution and recycling of the low density lipoprotein receptor as visualized with antireceptor antibodies. PMID- 6288728 TI - Effect of colchicine on the uptake of prolactin and insulin into Golgi fractions of rat liver. AB - In previous studies we have shown that 125I-labeled prolactin is taken up by a receptor-dependent process and concentrated in an intact form in Golgi elements from female rat liver (J. Biol. Chem., 1979, 254:209-214). In this study we have examined the effect of colchicine on this uptake process into Golgi elements. Colchicine [25 mumol (10 mg)/100 gm body wt] was injected intraperitoneally in adult female rats, and hepatic Golgi fractions were prepared at 1, 2, and 3 h postinjection. The enzyme recoveries and morphological appearance of fractions from colchicine-treated and control (alcohol alone) animals were similar. At times greater than 1 h after colchicine there was a marked (greater than 60%) inhibition of uptake of 125I-ovine prolactin (125I-oPRL) into Golgi light and intermediate fractions but no inhibition of uptake into Golgi heavy and plasmalemma elements. At times from 2 to 45 min postinjection, 125I-oPRL was extracted from Golgi elements and found to be largely intact as judged by rebinding to receptors. The inhibitory effect of colchicine was seen at doses ranging from 0.25 mumol to 25 mumol/100 g body wt. Vincristine also inhibited 125I-oPRL uptake into the Golgi light and intermediate fractions but lumicolchicine had no inhibitory effect. There was a smaller effect of colchicine both at early (1 h) and later (3 h) times on the extent and pattern of 125I insulin uptake. Colchicine treatment did not produce a significant change in lactogen receptor levels in the Golgi fractions. These results demonstrate that colchicine treatment inhibited the transfer of prolactin into Golgi vesicular elements. The much smaller effect on insulin uptake suggests that there may be differences in the manner in which the two hormones are handled in the course of internalization. PMID- 6288729 TI - Calcium transport of Plasmodium chabaudi-infected erythrocytes. AB - The calcium content and transport processes of Plasmodium chabaudi-infected rat erythrocytes were analyzed by atomic absorption spectroscopy and 45Ca2+ flux measurements. Infected erythrocytes, after fractionation on metrizamide gradients according to stage of parasite development, exhibited progressively increasing levels of Ca2+ with schizont and gametocytes containing 10- to 20-fold greater calcium levels than normal cells (0.54 +/- 0.25 nmol/10(8) cells). 45Ca2+ flux experiments showed both increased influx and decreased efflux in infected erythrocytes. Tris/NH4Cl lysis of normal erythrocytes preloaded with 45Ca2+ with the Ca2+ ionophore A23187 released less than 90% of cell calcium after incubation in ethyleneglycol bis(aminoethylether) N,N'-tetraacetic acid containing buffer, whereas lysis of the infected erythrocyte membrane resulted in release of 10-20% cell Ca2+, with the remaining portion associated with the isolated parasite fraction. This information together with the effects of various metabolic inhibitors indicates the presence of a parasite Ca2+ compartment in P. chabaudi infected erythrocytes. Dicyclohexylcarbodiimide (DCCD) an inhibitor of proton ATPases of chloroplasts, bacteria, yeast, and mitochondria, and the proton ionophore, carbonyl cyanide m-chlorophenylhydrazone (CCCP), inhibited Ca2+ influx and stimulated efflux from infected cells. These results combined with evidence for a DCCD- and CCCP-sensitive membrane potential in P. chabaudi-infected cells (Mikkelsen et al., accompanying manuscript) suggest that Ca2+ transport of intraerythrocytic parasites is coupled to a proton-motive force across the Plasmodia plasma membrane. PMID- 6288730 TI - Membrane potential of Plasmodium-infected erythrocytes. AB - The membrane potential (Em) of normal and Plasmodium chabaudi-infected rat erythrocytes was determined from the transmembrane distributions of the lipophilic anion, thiocyanate (SCN), and cation, triphenylmethylphosphonium (TPMP). The SCN- and TPMP-measured Em of normal erythrocytes are -6.5 +/- 3 mV and -10 +/- 4 mV, respectively. The TPMP-measured Em of infected cells depended on parasite developmental stage; "late" stages (schizonts and gametocytes) were characterized by a Em = -35 mV "early stages (ring and copurifying noninfected) by a low Em (-16 mV). The SCN-determined Em of infected cells was -7 mV regardless of parasite stage. Studies with different metabolic inhibitors including antimycin A, a proton ionophore (carbonylcyanide m chlorophenylhydrazone [CCCP] ), and a H+ -ATPase inhibitor (N,N' dicyclohexylcarbodiimide, [DCCD] ) indicate that SCN monitors the Em across the erythrocyte membrane of infected and normal cells whereas TPMP accumulation reflects the Em across the plasma membranes of both erythrocyte and parasite. These inhibitor studies also implicated proton fluxes in Em-generation of parasitized cells. Experiments with weak acids and bases to measure intracellular pH further support this proposal. Methylamine distribution and direct pH measurement after saponin lysis of erythrocyte membranes demonstrated an acidic pH for the erythrocyte matrix of infected cells. The transmembrane distributions of weak acids (acetate and 5,5-dimethyloxazolidine-2,4-dione) indicated a DCCD sensitive alkaline compartment. The combined results suggest that the intraerythrocyte parasite Em and delta pH are in part the consequence of an electrogenic proton pump localized to the parasite plasma membrane. PMID- 6288731 TI - Phospholipid metabolism, calcium flux, and the receptor-mediated induction of chemotaxis in rabbit neutrophils. AB - Rabbit neutrophils were stimulated with the chemotactic peptide fMet-Leu-Phe in the presence of the methyltransferase inhibitors homocysteine (HCYS) and 3 deazaadenosine (3-DZA). HCYS and 3-DZA inhibited chemotaxis, phospholipid methylation, and protein carboxymethylation in a dose-dependent manner. The chemotactic peptide-stimulated release of [14C]arachidonic acid previously incorporated into phospholipid was also partially blocked by the methyltransferase inhibitors. Stimulation by fMet-Leu-Phe or the calcium ionophore A23187 caused release of arachidonic acid but not of previously incorporated [14C]-labeled linoleic, oleic, or stearic acids. Unlike the arachidonic acid release caused by fMet-Leu-Phe, release stimulated by the ionophore could not be inhibited by HCYS and 3-DZA, suggesting that the release was caused by a different mechanism or by stimulating a step after methylation in the pathway from receptor activation to arachidonic acid release. Extracellular calcium was required for arachidonic acid release, and methyltransferase inhibitors were found to partially inhibit chemotactic peptide-stimulated calcium influx. These results suggest that methylation pathways may be associated with the chemotactic peptide receptor stimulation of calcium influx and activation of a phospholipase A2 specific for cleaving arachidonic acid from phospholipids. PMID- 6288732 TI - Direct cytochemical localization of catalytic subunits dissociated from cAMP dependent protein kinase in Reuber H-35 hepatoma cells. I. Development and validation of fluorescinated inhibitor. AB - A specific and sensitive procedure has been developed that reliably localizes intracellular sites of free catalytic unit (C) dissociated from cAMP-dependent protein kinase. The method is based on a FITC conjugate (F:PKI) of affinity column-purified heat-stable protein inhibitor (PKI) of free C. The fidelity of this cytochemical probe was determined using cultures of Reuber H-35 hepatoma cells that had been stimulated for 2 h with 0.1 mM DBcAMP, or with diluent, then fixed with anhydrous acetone at -30 degrees C. In these preparations the F:PKI probe complexed with free C in cytoplasm, nucleolus, and, to a minor extent, in nucleoplasm. Binding of the F:PKI molecule to free C was competitively diminished by arginine analogues, guanidinium HCI and polyarginine, each used over a 2-log dose range. When the inhibitor's arginine residues were blocked by reaction with cyclohexanedione it no longer inhibited phosphotransferase activity of free C, and when fluorescinated it failed to localize C in stimulated cells. Similarly, when F:PKI was preabsorbed with excess pure C it no longer functioned as a cytochemical stain. Affinity column-purified antibody to free C also reduced significantly the ability of F:PKI to complex with C in cell cultures stimulated with 0.1 mM DBcAMP. 1 microgram of antibody reduced by approximately 10% the binding of F:PKI to all cell compartments while 5 microgram of antibody diminished binding by greater than 50%. Together, these results indicate that the F:PKI binds specifically, perhaps exclusively, to the catalytic units of cAMP dependent protein kinase. The cytochemical procedure, unlike its biochemical counterparts, is able to locate the dissociation of cAMP-dependent protein kinase in individual cells of functionally or histologically complex cultures. Also, it reveals variations in the time- and dose-dependent activation of the kinase amongst clonal cells stimulated with cyclic nucleotide analogues or hormones. PMID- 6288734 TI - Synthesis of cartilage matrix by mammalian chondrocytes in vitro. I. Isolation, culture characteristics, and morphology. AB - We describe the isolation and the ultrastructural characteristics of adult bovine articular chondrocytes in vitro. Slices of bovine articular cartilage undergo sequential digestions with pronase and collagenase in order to release cells. Chondrocytes are plated at high density (1 x 10(5) cells/cm2) in culture dishes or roller bottles with Ham's F-12 medium, supplemented with 10% fetal bovine serum. Before culture, chondrocytes are freed of surrounding territorial matrix. Within the first few days of culture they re-establish a territorial matrix. As time progresses, chondrocytes synthesize both territorial and extraterritorial matrices. The matrices are rich in collagen fibrils and ruthenium red-positive proteoglycans. These features are most apparent in mass roller cultures in which aggregates of cells and matrix appear as long streaks and nodules. This morphology reveals an organization of chondrocytes and their matrices that is similar to that of the parent articular cartilage in vivo. PMID- 6288733 TI - Direct cytochemical localization of catalytic subunits dissociated from cAMP dependent protein kinase in Reuber H-35 hepatoma cells. II. Temporal and spatial kinetics. AB - The activation of cyclic AMP-dependent protein kinase has been found to be the predominant mode by which cyclic AMP (cAMP) leads to alterations of a large variety of cellular functions. The activation of the kinase results in the release of the catalytic subunit which as the free enzyme possesses phosphotransferase activity for a variety of specific protein substrates. Using a sensitive and specific cytofluorometric technique we monitored the appearance of free catalytic subunit in Reuber H35 hepatoma cells in culture after incubation with N6-1'-O-dibutyryl-cyclic AMP (DBcAMP), 8-bromoadenosine-3':5'-cyclic monophosphate (8-BrcAMP), and glucagon. The cytochemical method employs the heat stable inhibitor of the free catalytic subunit which has been conjugated to fluorescein isothiocyanate (F:PKI) and was validated as described in the companion paper (Fletcher and Byus. 1982. J. Cell Biol. 93:719-726). Here we studied the temporal and spatial kinetics of the free catalytic subunit following activation of cAMP-dependent protein kinase by increasing concentrations of DBcAMP,8-BrcAMP, and glucagon. Under similar conditions protein kinase activation was also assessed biochemically in H35 cell supernatants by assaying the protein kinase activity ratio. Incubation of the hepatoma cells with DBcAMP (0.1 mM) led to an increase in the activity ratio from 0.2 in control cultures to a value of nearly 1.0 within a 1- to 2-h period. During this same period using the F:PKI probe, a significant increase in cytoplasmic and nucleolar fluorescence indicative of the release of the free catalytic subunit was coincidentally observed. In contrast to the rapid appearance of catalytic subunit in the cytoplasm and nucleolus of the cell within 5-15 min of the addition of DBcAMP, discernible nucleoplasmic fluorescence did not occur until after 1 h. H35 cell cultures incubated with 8-BrcAMP (0.01-1.0 mM) exhibited a more rapid activation of the protein kinase measured cytochemically compared to the cells treated with DBcAMP. Cultures incubated with 8-BrcAMP had significantly increased cytoplasmic and nucleolar fluorescence compared to unstimulated cells within 1 min of the addition of the analogue and reached a maximal level within 15 min. By employing a microspectrophotometer a distinct dose-dependent increase in cellular fluorescence (i.e., free catalytic subunit) was observed as the concentration of 8-BrcAMP was increased from 0.01 to 1.0 mM at 1, 5, 15, and 60 min following stimulation. The addition of glucagon (10(-6) M) to the culture also led to the activation of cAMP-dependent protein kinase as determined by an increase in the activity ratio. This increase was paralleled throughout the incubation period by a marked elevation in cytoplasmic and nucleolar fluorescence. The results reported herein suggest that both cyclic nucleotide analogues and a polypeptide hormone lead to the activation of cAMP-dependent protein kinase in similar intracellular compartments in Reuber H35 hepatoma cells... PMID- 6288735 TI - Ca2+-sequestering smooth endoplasmic reticulum in an invertebrate photoreceptor. II. Its properties as revealed by microphotometric measurements. AB - Microphotometric measurements are used to investigate the functional properties of Ca2+-sequestering smooth endoplasmic reticulum (SER) in leech photoreceptors. 10-30 intact cells are mounted in a perfusion chamber, placed between crossed polarizers in a microphotometer, and permeabilized by saponin treatment. Subsequent perfusion with solutions containing Ca2+, MgATP, and oxalate leads to Ca uptake by SER. When the solubility product of Ca-oxalate is exceeded in the SER, birefringent Ca-oxalate precipitates form in the cisternae, leading to a large increase in the optical signal recorded from the preparation. The rate of increase in light intensity is used to measure the rate of Ca uptake. Ca uptake rate is linear with time over much of its course, can be switched on/off by the addition/withdrawal of Ca2+, ATP, or oxalate to/from the medium, and is inhibited by mersalyl and tetracaine. The Ca uptake mechanism has a high specificity for MgATP (KM,MgATP is approximately 0.8 mM). Uptake rates observed with dATP, GTP, UTP, ITP, and CTP are only 20-30% of the rate measured in ATP. The Ca pump has a high affinity for Ca2+ ions: the threshold for activation of the pump is approximately 5 x 10(-8) M, the apparent KM,Ca is approximately 4 x 10(-7) M. When Na+ or Li+ is substituted for K+, Ca uptake rate is decreased by 40-50%. The results show that the Ca2+-sequestering SER in leech photoreceptors shares some basic properties with skeletal muscle sarcoplasmic reticulum and supports the idea that certain subregions of the SER in invertebrate photoreceptors function as effective Ca2+ sinks/buffers close to the plasmalemma. PMID- 6288736 TI - Internalization and degradation of cholera toxin by cultured cells: relationship to toxin action. AB - Using anticholeragen antibodies and 125I-protein A, we developed a specific and quantitative assay for measuring choleragen on the surfaces of cultured cells. When neuroblastoma cells containing bound toxin were incubated at 37 degrees C, surface toxin disappeared with a half-life of approximately 2 h and a significant loss was detected by 10 min. When cells were incubated with 125I-choleragen in order to measure toxin degradation, cell-associated radioactivity disappeared with time and a corresponding amount of TCA-soluble label appeared in the culture medium with a half-life of 4-6 h. No degradation was detected until 45 min. Although there was a lag of 15 min before bound choleragen activated adenylate cyclase, the enzyme became maximally activated between 45 and 60 min. Similar results were obtained with Friend erythroleukemia cells. Internalization, degradation, and activation all were blocked when the cells were maintained at 4 degrees C. At 22 degrees C, internalization and activation occurred, albeit at a slower rate, whereas degradation was effectively inhibited. These results indicated that choleragen does not have to be degraded by intact cells in order for it to activate adenylate cyclase. Some internalization of the toxin, however, appears to precede the activation process. PMID- 6288737 TI - Transit of epidermal growth factor through coated pits of the Golgi system. AB - Using the direct conjugate of epidermal growth factor (EGF) and horseradish peroxidase, we have followed the entry of EGF into KB (human carcinoma) cells. EGF initially was found bound diffusely to the entire cell surface at 4 degrees C; on warming to 37 degrees C, EGF was found clustered in clathrin-coated pits on the plasma membrane in 1 min or less. Within 1-2 min at 37 degrees C, EGF began to accumulate in receptosomes within the cell and remained there for up to 10 min. At 10-13 min after warming to 37 degrees C, EGF was found in thin reticular membranous elements of the Golgi system, as well as concentrated in the clathrin coated pits present on these membranes. By 15 min after warming, EGF began to be delivered to lysosomes located near the Golgi system. These findings suggest that clathrin-coated pits in the Golgi reticular system accumulate EGF before delivery to lysosomes. PMID- 6288739 TI - Hormonally induced changes in the cytoskeleton of human thyroid cells in culture. AB - Serially cultivated thyroid follicular cells are not active in hormone synthesis but retain a thyrotropin-responsive adenylate cyclase. The exposure of such cells to thyrotropin leads to an increase in the concentration of intracellular cAMP and a drastic change in morphology including a total cytoplasmic arborization. The present communication describes these changes at the cytoskeletal level using a cell line derived from a human functioning thyroid adenoma. Phase contrast microscopy showed that the cytoplasmic arborization was preceded by a total disappearance of stress fibers, visible within 20 min of exposure. Small marginal membrane ruffles could also be seen. These morphological changes could also be induced by the addition of dibutyryl cAMP. The action of both thyrotropin and dibutyryl cAMP was potentiated by theophylline. High voltage electron microscopy of whole mounted cells confirmed the loss of stress fibers (microfilament bundles). In addition, thyrotropin treatment led to an uneven redistribution of the cytoplasmic ground substance and to changes in the organization of the microtrabecular lattice. Stereo images demonstrated numerous minute surface ruffles. The thyrotropin-induced arborization was reversible even in the presence of thyrotropin. After 24 h of treatment, cells had flattened and then contained very straight and condensed microfilament bundles. The results thus demonstrate that thyrotropin induces a disintegration of microfilament bundles in human, partially dedifferentiated, follicular cells and that this effect to all appearances is caused by cAMP, the second messenger in thyrotropin action. The relation of this event in partially dedifferentiated cells to the effect of thyrotropin in the intact thyroid gland is unclear. The fact that several other cultured hormone-responsive cells round up or become arborized in conjunction with an increase in cAMP levels implies that cAMP may be a major factor in the disassembly of microfilament bundles in these cells. PMID- 6288738 TI - Intracellular transport of the transmembrane glycoprotein G of vesicular stomatitis virus through the Golgi apparatus as visualized by electron microscope radioautography. AB - The intracellular migration of G protein in vesicular stomatitis virus-infected cells was visualized by light and electron microscope radioautography after a 2 min pulse with [3H]mannose followed by nonradioactive chase for various intervals. The radioactivity initially (at 5-10 min) appeared predominantly in the endoplasmic reticulum, and the [3H]mannose-labeled G protein produced was sensitive to endoglycosidase H. Silver grains were subsequently (at 30-40 min) observed over the Golgi apparatus, and the [3H]mannose-labeled G protein became resistant to endoglycosidase H digestion. Our data directly demonstrate the intracellular transport of a plasmalemma-destined transmembrane glycoprotein through the Golgi apparatus. PMID- 6288740 TI - Establishment of gonadotropin-responsive murine leydig tumor cell line. AB - Several clonal Leydig tumor cell lines have been established by adapting the transplantable Leydig tumor, M548OP, to culture. One of these cell line, MLTC-1, has been characterized with regard to the gonadotropin-responsive adenylate cyclase system. The binding of 125I-labeled human chorionic gonadotropin (hCG) was blocked by excess unlabeled hCG and lutropin (LH) but not by follitropin, thyrotropin, or insulin, indicating the presence of specific receptors for hCG and LH. Based on the specific binding of hCG to isolated MLTC-1 membranes, the calculated dissociation constant was 1.0 +/- 0.2 X 10(-10) M. The receptors appeared identical to those from normal murine Leydig cells when analyzed by SDS PAGE and sucrose density gradient centrifugation. The molecular weight and sedimentation coefficient were 95,000 daltons and 8.5 S, respectively. MLTC-1 cells responded to hCG by accumulating cyclic AMP and producing progesterone. Cyclic AMP accumulation was time- and dose-dependent with a maximal accumulation occurring at approximately 0.2 nM hCG. At saturating levels of hCG, cAMP levels reached a maximum by 30 min and then declined very slowly. Adenylate cyclase activity in membranes prepared from MLTC-1 cells was stimulated by hCG, LH, NaF, cholera toxin, and guanyl-5'-ylimidodiphosphate, Additionally, choleragen was found to ADP-ribosylate a membrane protein of 54,000 daltons. This protein resembles the proposed guanine nucleotide regulatory component in both size and choleragen-dependent reactivity. These data suggest that MLTC-1 cells possess a gonadotropin-responsive adenylate cyclase system consisting of a specific hormone receptor, a regulatory component, and a catalytic subunit. PMID- 6288741 TI - Volume regulation by human lymphocytes: characterization of the ionic basis for regulatory volume decrease. AB - The mechanism of volume regulation in hypotonic media was analysed in human peripheral blood mononuclear (PBM) cells. Electronic cell sizing showed that hypotonic swelling is followed by a regulatory volume decrease (RVD) phase. This was confirmed by both electron microscopy and by cellular water determinations. The rate of regulatory shrinking was proportional to the degree of hypotonicity in the 0.5-0.9 X isotonic range. Cell viability was only marginally affected in this range. The content of cellular K+ decreased during RVD, while Na+ content remained unchanged. Similarly, the efflux of 86Rb (used as a K+ analog) increased upon dilution, whereas 22Na efflux was not altered. 86Rb uptake was enhanced by hypotonic stress and both ouabain-sensitive and -insensitive components were affected. A ouabain-sensitive stimulation was also seen in Na+- free media. Ouabain partially inhibited RVD only if added to the cells hours before hypotonic challenge. A normal shrinking response was observed in K+-free media, and also in Na+-free media when Li+, choline+, or Tris+ were the substitutes. In high K+ or RB+ hypotonic media shrinking was absent and a second swelling phase was observed. Cs+ displayed an intermediate behavior, with shrinking observed at lower dilutions and secondary swelling at higher ones. The direction and magnitude of the response also changed when the external K+ concentration was varied and, with 50 mM K+, no regulatory volume change occurred following hypotonic stress. These findings suggest that RVD occurs largely by a passive loss of cellular K+, resulting from a selective increase in permeability to this ion. In addition, the (Na-K) pump appears to be activated upon cell swelling by a mechanism other than Na+ entry into the cell, but this activation is not essential for RVD. PMID- 6288742 TI - Distinct bone marrow precursors for mononuclear phagocytes expressing high and low 5' -nucleotidase activity. PMID- 6288743 TI - Synergistic stimulation of DNA synthesis by cyclic AMP derivatives and growth factors in mouse 3T3 cells. AB - Addition of the cAMP derivatives butcAMP or 8BrcAMP to quiescent cultures of Swiss 3T3 causes synergistic stimulation of DNAk synthesis with insulin, phorbol esters, vasopressin, epidermal growth factor, or fetal bovine serum (2-5%). In the presence of insulin, 8BrcAMP, and butcAMP stimulate [3H]-thymidine incorporation into acid-precipitable material in a dose-dependent manner. The effect of these agents is specific since 8Br5'AMP, 5'AMP, butyrate, or 8BrcGMP fail to stimulate DNA synthesis under identical experimental conditions. Furthermore, the mitogenic effects of the cAMP derivatives were markedly potentiated by 1-methyl-3-isobutyl xanthine and 4-(3-butoxy-4-methoxy benzyl)-2 imidazolidine, both of which are potent inhibitors of cyclic nucleotide phosphodiesterase activity. The growth-promoting effects of the cAMP derivatives were demonstrated by [3H]-thymidine incorporation (either by scintillation counting or by autoradiography), by flow cytofluorometric analysis, and by increase in cell number. When quiescent Swiss 3T3 cells were exposed to butcAMP and insulin, DNA synthesis began after a lag of 17h. The result of sequential additions of cAMP derivatives and insulin to quiescent 3T3 cells suggest that these agents must act simultaneously in G0/G1 to stimulate entry into DNA synthesis in these cells. The findings support the proposition that an increase in cellular levels of cAMP (but not cGMP) act sas a mitogenic stimulus for confluent and quiescent Swiss 3T3 cells. PMID- 6288744 TI - Effect of reduction of culture medium sodium, using different sodium chloride substitutes, on the proliferation of normal and Rous sarcoma virus-infected chicken fibroblasts. AB - We have substituted choline chloride, tetramethylammonium chloride, sucrose, or glucose for culture medium sodium chloride. When culture medium sodium is reduced below physiological levels (143 mM) by replacement of graded concentrations of sodium chloride with equivalent concentrations of choline chloride, normal fibroblasts approach proliferative inactivity in the presence of 90 mM Na, while their Rous sarcoma virus (RSV)-infected counterparts proliferate actively; both normal and neoplastic cells die with further sodium reduction. When culture medium NaC; is replaced with tetramethylammonium chloride, however, both normal and RSV-infected fibroblasts alike approach proliferative inactivity in the presence of 110 mM Na and both die off in the presence of 90 mM Na. When culture medium NaCl is replaced with sucrose or glucose yet another set of results is obtained: both normal and RSV-infected fibroblasts proliferate at reduced, although significant, rates in the presence of 42 mM Na. It is clear from our experimental results that the effects of reduction of culture medium sodium on cell proliferation differ markedly with the use of different sodium chloride substitutes. Caution must be exercised, therefore, in drawing inferences concerning the role of sodium in mitogenesis from experimental studies based on the tactic of reduction of external sodium. PMID- 6288745 TI - The use of fluorescent dyes to measure membrane potentials: a response. AB - The use of fluorescent cyanine dyes to estimate membrane potential in cell suspensions has been considered. Several problems related tot he application of the dyes have been reviewed. These problems include: 1) alteration of the membrane potential (Em) and factors involved in establishing Em by the dyes themselves, 2) the effects of altered energy metabolism on the fluorescent response of the dyes and on Em, and 3) calibration of dye fluorescence. Recent reports that advocate the use of the fluorescent dyes are misleading. PMID- 6288746 TI - A study of cAMP binding proteins on intact and disrupted sperm cells using 8 azidoadenosine 3',5'-cyclic monophosphate. AB - The photoaffinity probe (32P) 8-N3 cAMP was used to label the cAMP binding proteins in washed ejaculated human sperm. Three saturable binding proteins were photolabeled in both intact and disrupted cells with apparent molecular weights of 55,000, 49,000 and 40,000 daltons corresponding to the regulatory subunits of type II and type I cAMP-dependent protein kinase (cAMP-PK) and to an endogenous proteolytic product of the regulatory subunits, respectively. Photoincorporation in the three proteins could be totally blocked by preincubating the cells with cAMP. Cell-free seminal plasma was found to be free of detectable (32P) 8-N3 cAMP binding proteins. The 8-N, cAMP was also effective in stimulating endogenous cAMP PK activity in intact and disrupted sperm. A substantial amount of (32P) 8-N3 cAMP binding to types I and II regulatory subunits and cAMP-PK activity was detected on washed intact cells. Intact cells bound 1.80 pmol of (32P) 8-N3 cAMP/mg protein and had cAMP-PK activity of 824 units/10(8) cells. Disrupted cells bound 3.95 pmol (32P) 8-N3 cAMP/mg protein and had a cAMP-PK activity of 2,206 units/10(8) cells. The data presented support the concept of two classes of cAMP receptors being differentially available to externally added (32P) 8-N3 cAMP and proteases. Cellular membrane integrity and membrane sidedness are discussed as possible explanations for the observation reported. PMID- 6288747 TI - Immunological properties of gap junction protein from mouse liver. AB - Hepatic gap junctions were purified as plaques from BALB/c mice and separated by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate (SDS). Antisera were raised in rabbits and rats against gap junction plaques as well as protein bands of the following apparent molecular weights: 44K to 49K ("dimer" proteins), 26K, and 21K. Using an enzyme immunoassay, we found that the reactivities of the different antisera towards gap junction plaques decreased in the following order: anti-plaque antisera, anti-26K antisera, anti-"dimer" protein antisera, and anti-21K antisera. The gap junction protein bands separated by SDS-polyacrylamide gel electrophoresis were transferred by blotting onto nitrocellulose paper and the immunological cross-reactivities were compared: the anti-26K antisera reated with the dimer protein bands and the 26K band but did not cross-react with the 21K protein band. The rabbit anti-21K antiserum reacted weakly with the 21K protein. The missing immunological cross-reaction of the 26K and the 21K protein band can be most easily explained if both proteins were independent of each other. No inhibition of metabolic cooperation between fibroblastoid mouse 3T6 cells was observed in the presence of Fab fragments prepared from rabbit antiplaque antiserum or from rabbit anti 26K antiserum. When the total proteins of plasma membranes from mouse liver were separated by SDS polyacrylamide electrophoresis, only the 26K protein reacted with rabbit anti 26K antiserum. This result opens the possibility for direct quantitation of gap junction protein in tissues and cell fractions. PMID- 6288748 TI - Analysis of peptides in tissue and plasma. Use of silicic acid extraction and reversed-phase columns for rapid purification prior to radioimmunoassay. PMID- 6288749 TI - Direct isolation of beta-endorphin from plasma by column chromatography. PMID- 6288750 TI - Pituitary microadenomas causing Cushing's disease respond to corticotropin releasing factor. AB - Corticotropin-releasing factor (CRF) was administered as an iv bolus to two young women with mild Cushing's disease shortly before and one week after successful transsphenoidal microadenomectomy. The dose of CRF (1 microgram/kg body weight) had previously been shown to stimulate increased plasma ACTH and cortisol in normal subjects. In the first patient, prior to surgery, there were brisk increases in ACTH and cortisol that exceeded those observed in normal subjects. ACTH rose by 2 min and reached a peak between 15-30 min. Cortisol increased by 10 min and peaked between 45-60 min. After surgery, at a time when plasma cortisol was maintained at similar levels with exogenous hydrocortisone, there was no plasma ACTH or LH, TSH and prolactin increased after administration of LRH and TRH, and GH increased in response to insulin-induced hypoglycemia. The second patient had higher basal plasma ACTH and cortisol than the first patient. CRF induced increments in ACTH and cortisol were much less, but the time course was similar and peak levels attained were still higher than those in normal subjects. After surgery, at a time when plasma cortisol was maintained at a much lower level with exogenous hydrocortisone, there was no plasma ACTH or cortisol response. She had mild, transient diabetes insipidus. Basal levels of all other anterior pituitary hormones were normal. These results demonstrate that two microadenomas causing Cushing's disease were responsive to CRF in situ and suggest that CRF may be involved in the etiology and/or the responses to changes in plasma glucocorticoid concentrations observed in patients with Cushing's disease. PMID- 6288751 TI - Vitamin D resistant rickets with alopecia: cultured skin fibroblasts exhibit defective cytoplasmic receptors and unresponsiveness to 1,25(OH)2D3. AB - A new case of vitamin D dependent rickets (Type II) with alopecia in a 5 yr old child is reported. Skin fibroblasts were propagated in culture and analyzed for cytoplasmic 1,25(OH)2D3 receptors. The rachitic cells failed to exhibit specific, high-affinity binding sites by either Scatchard analysis or sucrose density gradient. Furthermore, a rise in 24-hydroxylase activity could not be elicited following incubation of the rachitig cells with 1,25(OH)2D3. Fibroblasts from a non-rachitic child examined in parallel experiments demonstrated high affinity binding sites (Kd = 0.1 nM, Nmax = 33 fmol/100 micrograms DNA) and the induction of 24-hydroxylase activity. The molecular basis of the unresponsiveness of the cells from the rachitic child appears to be due to defective or absent 1,25(OH)2D3 receptors. PMID- 6288752 TI - Metabolic clearance rate and plasma half-life of radioiodinated corticotropin releasing factor in a primate. AB - Corticotropin releasing factor (CRF) has recently been identified, purified and shown to be a 41-amino acid polypeptide that stimulates secretion of beta endorphin and ACTH. The metabolic clearance rates (MCR) of I125-CRF and I125 TyroCRF were measured in cynomologus monkeys using the pulse injection and the continuous infusion methods. Hunter-Bolton reagent was used for iodination of CRF and the chloramine-T method for Tyro-CRF. Gel chromatography was used to separate free iodine (and radioiodinated small fragments) from iodinated CRF in plasma samples. Disappearance of I125-CRF or I125-Tyro-CRF could be modeled with two components (bi-exponential). Plasma half-life of I125-CRF was 17.1 +/- 2.44 min (mean +/- SE, n=4) for the fast component and 198 +/- 5.3 min for the slow component. The MCR of I125-CRF using the pulse injection technique was 0.44 +/- 0.06 L/Kg/d, n=4 and the volume of distribution 213.5 +/- 12 ml, n=4, roughly equal to the plasma volume. Continuous infusion of I125-CRF and of I125-Tyro-CRF gave MCR's ranging between 2.23 -= 5.08 L/Kg/d, n=4. I125-ACTH MCR, measured for comparison using the same technique, was 5-10 fold higher. Thus, the plasma half life of CRF is longer than for all other known hypothalamic peptides and its metabolic clearance rate relatively low and considerably less all other known hypothalamic peptides and its metabolic clearance rate relatively low and considerably less than that of ACTH. Strong binding of CRF to plasma proteins may be responsible for the small volume of distribution and lower MCR values for the pulse injection method. PMID- 6288753 TI - Late-onset steroid 21-hydroxylase deficiency: a variant of classical congenital adrenal hyperplasia. AB - Hormonal studies and human leukocyte antigen (HLA) genotyping were performed in 5 males and 13 females who were demonstrated to have 21-hydroxylase deficiency. The enzymatic deficiency of steroidogenesis was detected by family studies of 10 females who presented with varying symptoms of androgen excess. The 10 index cases had normal genitalia at birth, but virilized to varying degrees postnatally. The additional 8 affected family members had not sought medical care, but some were found to have signs of virilization on physical examination, while others were normal. Thus both late-onset (symptomatic) and cryptic asymptomatic) 21-hydroxylase deficiency occurred in the same pedigree. The hormonal and genetic linkage studies indicate that the late-onset (symptomatic) form of 21-hydroxylase deficiency, like the cryptic (asymptomatic) and classical forms of 21-hydroxylase deficiency, is transmitted by an autosomal recessive gene which is linked to HLA-B. Furthermore, the classical form of 21-hydroxylase deficiency associated with prenatal virilization is transmitted by an allelic variant for steroid 21-hydroxylase different from that of the nonclassical forms, late-onset (symptomatic) and cryptic (asymptomatic) 21-hydroxylase deficiency. Although these latter 2 disorders have different clinical manifestations, they demonstrate a similar degree of steroid 21-hydroxylase deficiency that is less severe than that observed in classical 21-hydroxylase deficiency. The hormonal and genetic linkage data indicate that cryptic (asymptomatic) and late-onset (symptomatic) 21-hydroxylase deficiency result from the same allelic variant at the steroid 21-hydroxylase locus. A glossary of terms is presented to describe the various allelic forms of 21-hydroxylase deficiency with consistency. PMID- 6288754 TI - Attenuated forms of congenital adrenal hyperplasia due to 21-hydroxylase deficiency. AB - A variety of mild forms of congenital adrenal hyperplasia (CAH) due to partial 21 hydroxylase deficiency have recently been described. We report two families in whom members presented with CAH with various degrees of enzyme deficiency. In family A, two children had the classical salt-losing CAH. Their male sibling and mother presented a very mild asymptomatic form of CAH, characterized by elevated basal plasma levels of 17-hydroxyprogesterone (17-OHP) and exaggerated responses of progesterone and 17-OHP to ACTH stimulation. The Hormonal profile and HLA types of these two individuals suggested allelic compounds, having one mutant gene for classical CAH and another for a mild form. In family B, the proband presented an attenuated form of CAH, manifested by amenorrhea and hirsutism, elevated basal levels of plasma 17-OHP and androgens, as well as markedly increased ACTH response. Two of her four siblings had the same ad HLA type, elevated basal plasma 17-OHP levels, and increased ACTH response. Their father, their paternal aunt, and their paternal uncle had the ab HLA type and normal basal plasma 17-OHP but markedly increased ACTH response. The haplotypes a, b, and d were considered to be linked to a mutation resulting in mild 21-hydroxylase deficiency, the homozygotes with ab HLA type having a milder form of CAH than the homozygotes with the ad HLA type. The wide spectrum of clinical and hormonal characteristics among homozygotes for the 21-hydroxylase deficiency trait suggests that their is a continuum of degree of enzyme deficiency. Furthermore, it suggests that most nonclassical subjects are allelic compounds for variable degrees of severity in the mutation at the 21-hydroxylase locus. More specifically, the study of families A and B shows that the so-called cryptic and attenuated forms of CAH have the same pathophysiological basis. PMID- 6288755 TI - Effect of sex steroids on beta-endorphin in hypophyseal portal blood. AB - Previous studies in female monkeys have shown that beta-endorphin (beta-EP) of hypothalamic origin is present in high concentrations in the hypophyseal portal blood and declines at the time of menses and after ovariectomy. In this study we have examined the effects of estradiol and progesterone replacement on portal blood beta-EP in ovariectomized monkeys. After acute iv administration of estradiol (2 micrograms), beta-EP did not rise from previously low levels (less than 133 pg/ml) over the ensuing 3 h. After chronic estradiol replacement for 3 weeks, portal beta-EP was detectable in 2 of 4 ovariectomized monkeys, with peak values of 341 and 733 pg/ml, respectively. When progesterone as well as estradiol were replaced chronically, high levels of beta-EP, [1610 +/- 192 (SE) pg/ml] were measured in all 13 portal blood samples collected from 4 ovariectomized monkeys. The majority of the beta-EP immunoactivity in these samples eluted from a Sephadex G-50 column in the same position as synthetic human beta-EP. Cation exchange chromatography showed that the majority of immunoactive beta-EP in portal plasma appeared to be nonacetylated beta-EP (1-31). We conclude that ovarian steroids are necessary for the release of hypothalamic beta-EP into portal blood and suggest that cyclic changes in sex steroids may affect anterior pituitary function in part via a mechanism involving hypothalamic beta-EP. PMID- 6288757 TI - Concurrent production of adrenocorticotropin and prolactin from two distinct cell lines in a single pituitary adenoma: a detailed immunohistochemical analysis. AB - A pituitary tumor from a patient with severe Cushing's disease and marked hyperprolactinemia was extensively studied by immunohistochemical techniques. Tissues from two separate areas of the adenoma were found to contain similar cell proportions of PRL as well as ACTH and related peptides (beta-lipotropin, beta endorphin, and alpha MSH). The tumor was composed of approximately 70% immunoreactive PRL cells and 5% ACTH-containing cells. Double immunostaining revealed that PRL or ACTH and related peptides were found in two distinct populations of tumor cells. These results document for the first time inappropriate synthesis and secretion of an unusual combination of pituitary hormones from a mixed pituitary adenoma. PMID- 6288756 TI - Testicular luteinizing hormone receptor content and in vitro stimulation of cyclic adenosine 3',5'-monophosphate and steroid production: a comparison between man and rat. PMID- 6288758 TI - Angiotensin II receptors in human isolated renal glomeruli. AB - 125I-Labeled angiotensin II ([125I]A II) binds specifically to glomeruli isolated from human kidneys that were obtained at nephrectomy or early autopsy. Equilibrium was reached after 30 min, and specific binding represented more than 90% of the total binding. Dissociation after dilution with the addition of an excess of unlabeled hormone was more rapid than after dilution alone. The effect increased as a function of the A II concentration. The Scatchard plot derived from saturation experiments was curvilinear, with an upward concavity. Two groups of receptor sites could be defined by the Kd values (0.1 and 2 nM, respectively) and the number of receptor sites (40 and 300 fmol mg glomerular protein-1, respectively). Alternatively, binding could be considered to follow a negative cooperative type of hormone-receptor interaction. [Asn1, Val5]A II, [Asp1,Ile5]A II, [des, Asp1,Ile5]A II, [Sar1, Ala8]A II, and [Sar1, Ile8]A II were all equally effective as competitive inhibitors of [125I]A II binding. Both calcium and magnesium (0.5-5 mM) produced an increase in [125I]A II specific binding, whereas guanylylimidodiphosphate, an analog of GTP, inhibited it. Degradation of the [125I]A II present in the incubation medium was estimated by three different techniques. It increased linearly with time and reached 20% at 30 min. Specific binding of A II to human glomeruli at plasma concentrations observed in man under physiological conditions and during the iv administration of A II demonstrates that human renal glomeruli include target cells for A II and thus suggests a role for A II in regulation of the glomerular filtration rate in man. PMID- 6288760 TI - Phosphodiesterase activity in human subcutaneous adipose tissue in insulin- and noninsulin-dependent diabetes mellitus. AB - The influence of diabetes mellitus on phosphodiesterase (PDE) activity in human sc adipose tissue was investigated in 8 patients with insulin-dependent (IDDM) and 9 with noninsulin-dependent (NIDDM) diabetes mellitus. The results were compared with data from 10 healthy normal weight subjects. The apparent maximal PDE activity (Vmax) of the low Km form of PDE was 60% lower (P less than 0.01) in untreated IDDM and NIDDM than in the control state. After treatment of IDDM and NIDDM, the Vmax of the low Km PDE was normalized. In untreated IDDM and NIDDM, the Vmax of the low Km PDE was correlated to the cAMP level (r = 0.8). This correlation was not observed after antidiabetic treatment or in the control state. The apparent Vmax values of the high Km form of PDE were similar in the diabetic states and in control subjects. The results suggest that the low Km PDE is inhibited in untreated IDDM and NIDDM. In these conditions, PDE may be one factor responsible for regulation of the cAMP level. PMID- 6288759 TI - Vasoactive intestinal peptide stimulates adrenocorticotropin release from human corticotropinoma cells in culture: interaction with arginine vasopressin and hydrocortisone. AB - The effect of vasoactive intestinal peptide (VIP), cholecystokinin octapeptide (CCK), bombesin, arginine vasopressin (AVP), and hydrocortisone (HC) on ACTH release from human corticotropinoma cells in culture has been studied. Tumor tissue was obtained from 6 patients with pituitary corticotropinomas. Eleven to 21 cultures yielding 0.7-2.0 X 10(6) cells/culture, were obtained from each tumor and maintained for periods of 4 weeks to longer than 6 months. VIP (500 ng/ml) significantly (P less than 0.005) stimulated ACTH release from all tumors studied, and a dose (5-500 ng/ml)-response effect was observed in 3 of 5 tumors. Stimulation by VIP was seen at 2,4, and 24 h and was maximal at 4 h. CCK and bombesin were without effect on ACTH release from 4 tumors studies at 4 h. AVP (1 10 mU/ml) stimulated ACTH from 4 tumors studied at 60 min or 4 h. Coincubation of cultures with VIP (50-500 ng/ml) and AVP (1-10 mU/ml) resulted in at least an additive effect. HC (100 ng/ml) significantly (P less than 0.025) inhibited basal ACTH secretion from 2 of 4 tumors at 4 h and from 3 of 4 (P less than 0.005) at 24 h. Simultaneous coincubation of cultures with VIP (50 ng/ml) and HC (100 ng/ml) resulted in an attenuation or blockade of the VIP-stimulated ACTH release, whereas prior incubation of cultures with HC for 28 h before exposure to VIP did not. The results demonstrate that VIP is a potent ACTH secretagogue from human corticotropinoma cells in culture; its effects are additive to those of AVP and modulated by HC. PMID- 6288761 TI - In vitro studies with cefotaxime: disk diffusion susceptibility tests. AB - Until recently two sets of conflicting interpretive criteria existed for use with the standardized 30-mug cefotaxime diffusion disk [National Committee for Clinical Laboratory Standards (NCCLS) M2-A2S, supplement 1; product information package insert for Claforan (cefotaxime sodium), edition 10/81.] The latter criteria recently were superseded by a third set which differs radically from the first two in both minimal inhibitory concentration (MIC) and zone size breakpoints. The first two sets of criteria differed mainly in the zone of inhibition diameters used to predict resistant and intermediate organisms. The accuracies of these two sets of criteria were evaluated by paired agar dilution (World Health Organization-International Collaborative Study) and disk diffusion tests (Food and Drug Administration) with 347 clinical isolates of aerobic bacteria. A total of 274 isolates (79%) were clinically susceptible by agar dilution as defined by NCCLS (MIC /=64 mug/ml). The original product information package insert criteria proved to be most unreliable in identification of the intermediate organisms (zone of inhibition diameters of 18 to 22 mm); only 5 were correctly predicted as being intermediate, whereas 54 were predicted as being resistant, and 2 were predicted as being susceptible. In contrast, the NCCLS criteria (zone of inhibition diameter of 15 to 22 mm) predicted 41 as being intermediate and 18 as being resistant; again, 2 were susceptible. The 12 isolates resistant to cefotaxime by agar dilution were correctly predicted to be resistant by either set of breakpoints (zone of inhibition diameter of 5 mM and arterial pH <7.25. In addition to supportive care, treatment usually consists of intravenous NaHCO(3), with a resultant mortality >60%. Dichloroacetate (DCA) is a compound that lowers blood lactate levels under various conditions in both man and laboratory animals. It acts to increase pyruvate oxidation by activation of pyruvate dehydrogenase. We evaluated the effects of DCA in the treatment of two different models of type B experimental lactic acidosis in diabetic dogs: hepatectomy-lactic acidosis and phenformin-lactic acidosis. The metabolic and systemic effects examined included arterial blood pH and levels of bicarbonate and lactate; the intracellular pH (pHi) in liver and skeletal muscle; cardiac index, arterial blood pressure and liver blood flow; liver lactate uptake and extrahepatic splanchnic (gut) lactate production; and mortality. Effects of DCA were compared with those of either NaCl or NaHCO(3). The infusion of DCA and NaHCO(3), delivered equal amounts of volume and sodium, although the quantity of NaHCO(3) infused (2.5 meq/kg per h) was insufficient to normalize arterial pH. In phenformin-lactic acidosis, DCA-treated animals had a mortality of 22%, vs. 89% in those treated with NaHCO(3). DCA therapy increased arterial pH and bicarbonate, liver pHi and cardiac index, with increased liver lactate uptake and a fall in blood lactate. With NaHCO(3) therapy, there were decrements of cardiac index and liver pHi, with an increase in venous pCO(2) and gut production of lactate. Dogs with hepatectomy-lactic acidosis were either treated or pretreated with DCA. Treatment with DCA resulted in stabilization of cardiac index, a fall in blood lactate, and 17% mortality. NaHCO(3) was associated with a continuous decline of cardiac index, rise in blood lactate, and 67% mortality. In dogs pretreated with NaCl, mortality was 33%, but all dogs pretreated with DCA survived. Dogs pretreated with DCA also had lower blood lactate and higher arterial pH and bicarbonate than did those pretreated with NaCl.Thus, in either of two models of type B experimental lactic acidosis, treatment with DCA improves cardiac index, arterial pH, bicarbonate and lactate, and liver pHi. The mortality in dogs with type B lactic acidosis was significantly less in DCA-treated animals than in those treated with other modalities. PMID- 6288775 TI - Malignant fibrous histiocytomas of salivary glands. AB - The light microscopic, immunohistological and ultrastructural findings in two cases of malignant fibrous histiocytoma arising in salivary glands are presented and the features of seven previously reported cases are reviewed. This neoplasm is extremely rare in this site and may pose problems in diagnosis. It has to be distinguished from other spindled cell tumours, in particular from epithelial tumours of predominantly spindled cell pattern; immunohistological markers for histiocytic cells may be of value. The histogenesis of this neoplasm is controversial but our electron microscopic findings support an origin from mesenchymal cells which differentiate along a broad fibrohistiocytic spectrum. PMID- 6288774 TI - Complement biosynthesis by the human hepatoma-derived cell line HepG2. AB - The human hepatoma-derived cell line, HepG2, synthesized and secreted functional complement proteins C1r, C1s, C2, C3, C4, C5, factor B, C1 inhibitor, C3b inactivator, a small amount of C6, and trace amounts of C8; but failed to produce detectable C1q, C7, or C9. Immunochemically, C2, C3, C4, C5, and B were isolated from culture medium as proteins with molecular sizes and subunit structures identical to the corresponding components isolated from serum. C2 and factor B from cellular lysates had slightly lower molecular weights than the corresponding proteins in culture medium. C3, C4, and C5 were detected as single chain precursor molecules in cellular lysates. These results demonstrate that human C5, like C3 and C4, is synthesized as a single chain precursor that is converted by limited proteolysis to the native two-chain molecule. It also establishes the precursor-product relationship for human pro-C4 and native C4, pro-C5, and native C5. PMID- 6288776 TI - Synaptic complexes formed by functionally defined primary afferent units with fine myelinated fibers. AB - The individual fine myelinated fibers of cutaneous mechanical nociceptors and "D hair" receptors were identified by electrophysiological recording with micropipette electrodes in cats and monkeys. Their intraspinal terminations were labeled by iontophoresing horseradish peroxidase intracellularly and subsequent diaminobenzidine histochemistry. These terminations were examined with light and electron microscopy to determine the nature and organization of their synaptic contacts. Myelinated fibers of the mechanical nociceptors became unmyelinated before exhibiting many enlargements that made multiple synaptic contacts in the marginal zone (lamina I) and lamina V. Pre- or postsynaptic contacts were found only on enlargements. In the marginal zone of the cat, enlargements made simple axodendritic contacts or were scalloped, central terminals in glomeruli. In glomeruli, myelinated mechanical nociceptor enlargements were presynaptic to several dendritic appendages and postsynaptic to two different types of profiles. One type was interpreted as a presynaptic axon terminal, the other as a presynaptic, vesicle-containing, dendritic appendage. In lamina V of the cat the nociceptor synaptic complexes were similar, but simpler, and only axonal profiles were found to be presynaptic to them. In the monkey marginal zone and deep nucleus proprius, myelinated nociceptor terminations formed the central element of glomeruli, which consisted of postsynaptic dendritic appendages and presynaptic axon terminals. D-hair axons terminated in large numbers of enlargements in the nucleus proprius (laminae III and IV) and inner substantia gelatinosa (lamina IIi). Their large rounded enlargements formed the central terminals in glomeruli and were presynaptic to both ordinary and vesicle containing dendritic appendages; the presynaptic dendritic profiles also often contacted each other. Profiles interpreted as axonal in origin were the only terminals presynaptic to the primary ending within the D-hair glomeruli. The results suggest that transfer of primary afferent information occurs only at enlargements of the primary fiber and that each primary fiber enters into more than one kind of synaptic arrangement. They also point out that synaptic glomeruli are common to functionally different types of primary afferent fibers and that the internal organization of glomeruli varies with the kind of primary fiber and the locus of the complex. PMID- 6288777 TI - A physiological and morphological study of the horizontal cell types of the rabbit retina. AB - A perfused, isolated retina-eyecup preparation of the rabbit was utilized to correlate the physiology and morphology of horizontal cells. Neurons were physiologically characterized by intracellular recording techniques and subsequently stained with intracellular iontophoretically injected horseradish peroxidase for morphological identification. Three types of rabbit horizontal cell recordings have been differentiated, based on variations in response waveform, amplitude-intensity properties, and area summation characteristics. These three types have been unequivocally associated with the axonless A-type horizontal cells and the somatic and axon terminal endings (each displaying its own distinct physiology) of B-type horizontal cells first described in studies using Golgi-impregnation techniques (Fisher and Boycott, '74). In addition, the sizes of A-type horizontal cells were found to be directly related to their retinal eccentricities from the optic desk. However a unique subclass of A-type cells has been discovered (elongated or Ae type) which displayed the largest dendritic field of any cells studied here, yet had the smallest eccentricities- within 1.4 mm of the optic disk. Moreover, elongated A-type cells exhibited long asymmetrical dendritic fields which were oriented parallel with the visual streak. The unique asymmetry and orientation of these cells suggests that they may have orientation-biased receptive field properties. Physiological evidence for an orientation-biased horizontal cell is presented in support of this notion. PMID- 6288778 TI - Flavor-illness aversions: potentiation of odor by taste with toxin but not shock in rats. AB - Potentiation of odor by taste in rats was tested in a variety of situations. In three experiments, almond odor and saccharin taste were presented either as a single conditioned stimulus (CS) or as a compound CS and followed by either toxic lithium chloride or footshock. Extinction tests with the almond and saccharin components were then given. In single CS-toxin experiments, taste was more effective than odor, and after compound conditioning, the taste component potentiated the odor component. Conversely, in single CS-shock experiments, odor was more effective than taste, and after compound conditioning, no potentiation was observed. Rather, interference effects were observed. In Experiments 1 and 2, the addition of taste disrupted odor CS-shock conditioning, and in Experiment 3, odor interfered with taste CS-shock conditioning. Visceral feedback is apparently a necessary unconditioned stimulus for the potentiation of odor by taste. These data support the neural convergence and gating hypothesis of flavor aversion conditioning. PMID- 6288780 TI - Long-delay associations between drug states produced in rats by injecting two drugs in sequence. AB - Injection of pentobarbital after a rat has consumed saccharin solution usually produces a mild aversion to the saccharin. However, the pentobarbital-produced aversion is eliminated or attenuated by prior pairings of pentobarbital injections with lithium injections. This is called the Avfail (aversion failure) effect. The present experiments dealt with the effect of the temporal relation of the pentobarbital injection to the lithium injection. After forward pairings (pentobarbital before lithium) with delays between the two injections varying among groups from 2.5 min to 320 min, Avfail was invariably obtained. There was little effect of the length of the forward delay, although the Avfail effect appeared to be slightly stronger at 30-40 min or so. When the two drugs were injected simultaneously or in a backward sequence, there was a weakening of the flavor aversion produced by pentobarbital, but this is attributable to habituation to the drugs, not to Avfail. PMID- 6288779 TI - Flavor-illness aversions: gustatory neocortex ablations disrupt taste but not taste-potentiated odor cues. AB - Two experiments evaluated the contribution of the gustatory neocortex (GN) to the potentiation of odor by taste during illness-induced aversions in rats. In Experiment 1, rats lacking GN and control rats were given an odor, a taste, or an odor-taste compound cue followed by intragastric gavage of lithium chloride. Prior to conditioning, neophobia for flavored solutions was absent in rats with GN lesions. After pairing with LiCl, GN rats developed normal conditioned odor aversions (Experiment 1B), whereas conditioned taste aversions were attenuated (Experiment 1A) or totally blocked (Experiment 1B). Potentiation of odor by taste after compound conditioning was evident in both control and GN rats, although GN lesions attenuated the effect slightly in Experiment 1B. In Experiment 2, normal rats were given compound conditioning to induce potentiated odor aversions and then given GN lesions prior to tests with the odor and taste components. Taste aversion retention was disrupted totally by GN ablation; potentiated odor aversions were retained by both groups, although the GN group extinguished faster. Gustatory neocortex ablations produced differential effects on odor and taste, disrupting taste memorial and associative processes but leaving odor conditioning and the potentiation of odor by taste processes relatively unaffected. Integrity of the GN apparently is not necessary for the acquisition or retention of potentiation odor aversions. PMID- 6288781 TI - Computed tomography of bronchial adenomas. AB - Computed tomography (CT) was used to evaluate nine patients with bronchial adenomas: five carcinoid tumors and four adenoid-cystic carcinomas. Seven patients with lesions of the trachea or proximal bronchi presented with wheezing, hemoptysis, or obstructive pneumonitis. In such cases CT was effective in displaying the total extent of infiltrating lesions including the extraluminal component. The cases of adenoid-cystic carcinoma of the bronchi demonstrated extraluminal spread in a pattern indistinguishable from bronchogenic carcinoma. In one patient, CT demonstrated an exclusively endobronchial carcinoid tumor, and conservative resection was planned and accomplished. In two cases of carcinoid tumors that presented as solitary pulmonary nodules adjacent to bronchi in the midlung zone, CT served as a road map for the bronchoscopist. Computed tomography has been employed in the post-operative patient to evaluate for possible recurrence. PMID- 6288782 TI - Computed tomography and routine chest radiography in oat cell carcinoma of the lung. AB - Computed tomography (CT) scans of the chest and routine postero-anterior and lateral chest films were compared in 32 patients with pathologically proven oat cell carcinoma of the lung. Nineteen of the 32 patients were examined prior to any form of therapy. Overall, CT gave more information about the extent of disease. The significance of this information may be of little value because the routine chest films also demonstrated mediastinal involvement, although to a lesser degree. Computed tomography of the chest should not be the initial diagnostic staging procedure in oat cell carcinoma, but used selectively. It can be quite useful in patients who have a partially opacified lung, possibly for the evaluation of mediastinum after therapy, and for the evaluation of possible abdominal metastases. PMID- 6288783 TI - Pancreatic cysts caused by carcinoma of the pancreas: a pitfall in the diagnosis of pancreatic carcinoma. AB - Pancreatic cysts were diagnosed on computed tomography (CT) in 6 (8%) of 73 histologically proven pancreatic carcinomas. In three of a total of seven cases studied, the initial CT diagnosis was pancreatitis. The CT features of cysts secondary to pancreatic carcinoma were identical to those of pseudocysts in pancreatitis. All cases had masses of varying sizes suggestive of carcinoma or pancreatitis. Even in retrospect, two cases were difficult to differentiate from pancreatitis with pseudocysts. Our results indicate that whenever pancreatic cysts are encountered in patients of cancer age without a history of pancreatitis, further examinations including percutaneous aspiration biopsies should be performed to exclude malignancy. PMID- 6288784 TI - Computed tomography in malignant fibrous histiocytoma. AB - The computed tomographic findings in 11 cases of malignant fibrous histiocytoma are discussed. The typical appearance of the eight primary tumors imaged was of a poorly marginated mass with an attenuation coefficient slightly lower than that of normal muscle. Central tumor necrosis was detected in three cases. Three of five abdominal tumors showed involvement of adjacent soft tissue structures, and one demonstrated destruction of adjacent bone. The nonspecific appearance of these tumors prevented their reliable differentiation from other soft tissue sarcomata. PMID- 6288785 TI - Hepatocellular carcinoma involving the portal vein. AB - A case of hepatocellular carcinoma with invasion of the portal vein is reported. A tumor defect within the dilated portal vein is well demonstrated by computed tomography, and its appearance is compatible with the "thread and streaks" sign seen on hepatic angiography. PMID- 6288787 TI - Effect of fiber in corn tortillas and cooked beans on iron availability. PMID- 6288786 TI - 2-Haloacrylic acids as indicators of mutagenic 2-haloacrolein intermediates in mammalian metabolism of selected promutagens and carcinogens. PMID- 6288788 TI - A clinical comparison of a quartz- and glass-filled composite with a glass-filled composite. PMID- 6288789 TI - Development and loss of toluene diisocyanate reactivity: immunologic, pharmacologic, and provocative challenge studies. AB - Toluene diisocyanate (TDI) sensitivity accompanied by nonspecific bronchial hyperresponsiveness occurs in approximately 5% of occupationally exposed workers. We report the case of a 32-yr-old worker followed longitudinally after removal from isocyanate exposure. TDI reactivity was lost 11 mo after removal from exposure and nonspecific bronchial hyperresponsiveness resolved after 17 mo. Bronchial reactivity to radishes (Raphanus sativus), which developed concurrently with TDI reactivity, was lost 2 yr later. Immunopharmacologic results show that the worker's initial decreased ability of lymphocytes to produce cyclic AMP returned to near normal after 2 yr. IgE antibodies to a human serum albumin tolyl monoisocyanate conjugate were still present at this time. PMID- 6288790 TI - The beta-adrenergic system and allergic bronchial asthma: changes in lymphocyte beta-adrenergic receptor number and adenylate cyclase activity after an allergen induced asthmatic attack. AB - Beta-adrenergic receptor characteristics and adenylate cyclase responses to different stimuli were measured in lymphocyte membrane preparations of six normal control subjects and five allergic asthmatic patients with mild airways disease and increased bronchial reactivity to histamine and acetylcholine. The determinations were performed just before and 24 hr after inhalation challenge with house-dust mite antigen to investigate the influence of an allergen-induced asthmatic attack on the beta-adrenergic receptor system. Before the house-dust mite challenge, the lymphocyte membranes of the patients showed a normal receptor density, dissociation constant for (-)3H-dihydroalprenolol, and adenylate cyclase response to isoproterenol, guanyl-5'-yl-imidodiphosphate, (GppNHp) and NaF. After the allergen-induced asthmatic reaction, however, the beta-adrenergic receptor number in the patients was significantly reduced by 21%, while the basal adenylate cyclase activity and isoproterenol-, GppNHp-, and NaF-induced cyclic AMP responses were simultaneously reduced by about 40%. The allergen challenge had no significant effect on the lymphocyte membranes of the control subjects. These results suggest (1) that a reduced beta-adrenergic function is not an intrinsic component of allergic bronchial asthma but is rather the consequence of an active disease state, and (2) that next to changes in beta-adrenergic receptor number, alterations distal to the receptor may play an important role in the observed decrease in beta-adrenergic responsiveness. PMID- 6288791 TI - Adrenergic receptors and increased reactivity of aortic smooth muscle in renal hypertensive rats. AB - The contractile responses of aortic smooth muscle and the dissociation constant (Ka) of the norepinephrine (NE) acting on alpha-adrenergic receptors in isolated thoracic aorta from normotensive and renal hypertensive rats were studied. Male Wistar rats were made hypertensive by uninephrectomy with figure-of-eight ligatures around the contralateral kidney. Two to three weeks after operation, the arterial systolic pressure for sham-operated rats was 123 +/- 2, and that for hypertensive rats was 164 +/- 4 mm Hg. Spontaneous rhythmic contractions of aortic rings, which were potentiated by low [Na]o and abolished by nifedipine (10(-6) - 10(-5) M), were observed in 4 out of 18 hypertensive rats. Aortic rings from normotensive rats showed no spontaneous rhythmicity. Although rhythmic phasic contractions could be induced in normotensive tissues by NE or caffeine in the presence of NE, tissues of hypertensive rats responded more readily. Furthermore, the threshold and the ED50 for tonic contractile response to NE was lower in hypertensive aortic rings than in the rings obtained from normotensive aortas. The Ka of NE acting on alpha-adrenergic receptors was derived from concentration-response data for NE before and after irreversible blockade of a fraction of the receptors with dibenamine (1-2 X 10(-7) M) in the presence of cocaine and propranolol. Mean Kas obtained were 5.58 +/- 0.42 X 10(-7) and 2.12 +/- 0.28 X 10(-7) M for normotensive and hypertensive rats, respectively. The increased contractile responses of the hypertensive aorta may be explained, at least partially, by a higher resting [Ca]i and a greater affinity of NE acting on the alpha-adrenergic receptors. PMID- 6288793 TI - Effects of cortisol and ACTH on adrenocortical growth and cytodifferentiation in the hypophysectomized fetal sheep. PMID- 6288792 TI - Effects of dopaminergic agonists on somato-autonomic reflexes. AB - In cats lightly anesthetized with urethane (600 mg/kg, i.p.) reflexes of the blood pressure (BP) and of the nictitating membrane (NM) were elicited by stimulation of the sciatic nerve (16 V, 0.3 ms, 1-128 Hz, 2 s or 2 min) prior to and after the administration of apomorphine (0.05-0.2 mg/kg, i.v.) or piribedil (0.4-1.0 mg/kg, i.v.). In case of short-train (2 s) stimulation, both dopaminergic agonists shifted the frequency-response curves of NM contractions to the right, i.e. depressed NM reflexes in the entire range of the stimulation frequencies applied. At the same time, BP reflexes were depressed only in the range of lower frequencies (1-4 Hz). At higher rates (32-128 Hz) BP reflexes were potentiated. The reactions of BP to sustained (2 min) stimulations displayed a flat pressor plateau in response to lower-frequency stimulation, and a two component pattern (an initial pressor peak followed by a plateau) to the higher frequency one. Compatibility with the effects seen to short-train stimulations, the dopaminergic agonists prolonged the rise-time and augmented the amplitude of the initial pressor peak to sustained stimulations with lower and higher frequencies, respectively. The plateau of the pressor response to higher frequencies was depressed by higher doses (greater than 0.4 mg/kg) of piribedil. Administration of haloperidol (0.05-0.2 mg/kg, i.v.) resulted only in a partial restoration of the reflexes of BP and NM. The manifold effects of dopaminergic agonists on the somato-autonomic reflexes studied support the thought than NM and BP reflexes are organized, at least partially, in different ways. PMID- 6288794 TI - Froment's paper sign and Jeanne's sign-- unusual etiology. PMID- 6288795 TI - Granular cell tumor of the esophagus: a case report. PMID- 6288796 TI - Virulence of entamoeba histolytica strains from Bombay (India) to rats. AB - The results of intracaecal inoculation experiments on young albino rats with different strains of E. histolytica isolated in cultures from patients with different varieties of clinical amoebiasis, such as acute amoebic dysentery, amoebic liver abscess and chronic amoebic colitis, as well as from asymptomatic infections, have been presented and discussed. Caecal scoring was performed with regard to the condition of the wall of the caecum and the contents of its lumen. Successful infection with virulent amoebae was associated with the presence of considerable ulceration of the caecum, whereas, in infections with avirulent amoebae, the changes were apparently only slight or absent. The virulence and pathogenicity were considerably different for clinical case and symptomless carrier strains of E. histolytica. In the former group, high caecal scores and pathogenicity indices with ulceration of the caecum were noted, whilst in the latter, the amoebae were either virulent or avirulent. Results similar to those for the former group were obtained with two carrier strains only. PMID- 6288797 TI - Properties of poliovirus strains isolated over the period 1969-1978 from the main municipal sewerage system of the city of Prague. AB - Biological properties (rct marker and antigenic relatedness) were compared in vaccine prototype strains and in 62 poliovirus strains isolated during a period 1969-1978 from the main municipal sewerage system of the City of Prague. None of the strains isolated from the municipal sewerage showed biologic properties that would fully differ from those observed in vaccine-derived strains. The strains detected very late postvaccination (after about a year) showed a lesser extent of changes than strains isolated earlier after vaccination. The most frequent changes were recorded in type 2 strains, less frequent in type 3 strains and the least frequency of changes was found in type 1 strains. To facilitate comparisons of these changes in dependence on time of postvaccination virus excretion a supporting evaluation criterion has been developed to help express the dynamics of changes in the isolated poliovirus strains. The recorded degree of the dynamics of changes was highest in type 2 strains, lower in type 3 strains and lowest in type 1 strains. The dynamics of changes detected in strains of various types was not always constant in the course of years: in a given year (or in a period of several years) changes occurred always in strains of the same serologic type, whereas strains of the other two types changed only insignificantly during the respective period. PMID- 6288798 TI - Regeneration of T cell subpopulations after bone marrow transplantation: cytomegalovirus infection and lymphoid subset imbalance. AB - Immune reconstitution was studied serially by using T lymphocyte cell surface differentiation antigens in 37 individuals receiving bone marrow transplants. Antigen expression was assessed by immunofluorescence analysis using monoclonal antibodies to T lymphocytes including Leu-3, which defines a T lymphocyte subpopulation in healthy individuals with T helper or inducer activity (L3+), and Leu-2, which defines a T lymphocyte subpopulation in healthy individuals with T helper or inducer activity (L3+), and Leu-2, which defines a T lymphocyte subpopulation with suppressor or cytotoxic activity (L2+). These studies demonstrated that the L2+ subpopulation regenerated more rapidly after transplant than did the L3+ subpopulation. Imbalances between these two T lymphocyte subpopulations, indicated by a decreased L3/L2 ratio, persisted for periods up to 12 mo post-transplant. Expression of a cell surface antigen associated with immature lymphocytes (OKT-10), and of HLA-DR (Ia-like) antigens was markedly increased during the post-transplant period. HLA-DR antigen expression did not appear related to immune activation in that increased reactivity was not detected with a monoclonal antibody (anti-TAC) specific for activated T cells. These observations in bone marrow transplant recipients and other disorders characterized by lymphoid restoration or immaturity indicate that inversion of the normal L3/L2 ratio and increased expression of OKT-10 and HLA-DR antigens may be features of a regenerating immune system. Furthermore, serial observation of individual patients indicated that infection with cytomegalovirus was associated with a progressive decrease in the L3/L2 ratio. PMID- 6288799 TI - Alteration of in vitro immunoglobulin secretion by amosite asbestos. AB - We investigated the in vitro effects of amosite asbestos on immunoglobulin (Ig) secretion by human peripheral blood mononuclear leukocytes (MNL). Concentrations of 100 to 300 micrograms/ml of amosite asbestos reduced the number of Ig secreting cells recovered from 6-day cultures of unstimulated MNL or MNL stimulated with Epstein Barr virus. By contrast, the Ig secretory response to pokeweed mitogen was enhanced by 10 to 100 micrograms/ml concentrations of amosite asbestos; however, amosite asbestos no longer enhanced the response to pokeweed mitogen when MNL were first partially depleted of monocytes (to less than 2%) esterase-positive cells remaining). These results indicate that amosite asbestos has multiple effects on the cells involved in Ig secretion: 1) amosite asbestos inhibits unstimulated B cell function; 2) amosite asbestos inhibits the function of B cells stimulated with the direct B cell activator Epstein Barr virus; and 3) amosite asbestos may alter regulator monocyte function allowing enhanced Ig secretion in the presence of monocyte-dependent B cell triggers such as pokeweed mitogen. PMID- 6288800 TI - Selective induction of autoantibody secretion in human bone marrow by Epstein Barr virus. AB - The bone marrow is an important site for B lymphocyte differentiation and antibody synthesis in animal and man. However, few experiments have examined directly its immunologic functions in humans. In the present experiments, we have induced bone marrow B lymphocytes from human donors with degenerative arthritis of varying ages to secrete two autoantibodies, IgM and anti-IgG (rheumatoid factor) and IgM anti-human thyroglobulin (Tg), by stimulation with the polyclonal B cell activator Epstein Barr virus (EBV). The EBV-stimulated bone marrow cells secreted significantly more IgM anti-IgG (p less than 0.01) and IgG anti-Tg (p less than 0.01) than matched, identically treated peripheral blood cells. Bone marrow cultures from donors over the age of 60 yr, particularly females, produced more rheumatoid factor than cultures from younger donors (p less than 0.01). The EBV-inducible autoantibodies were immunospecific as demonstrated by adsorption studies. A potential pathogenic role in the inflammatory process was suggested by the finding that the EBV-inducible IgM anti-IgG autoantibodies were capable of activating the classical complement pathway as assessed by the cleavage of C4. These results indicate that the human bone marrow is a selective reservoir for EBV-inducible autoantibody precursor B lymphocytes, and that the size of the reservoir increases with age. PMID- 6288801 TI - Susceptibility of Legionella pneumophila to oxygen-dependent microbicidal systems. PMID- 6288802 TI - Host immune response to reovirus: CTL recognize the major neutralization domain of the viral hemagglutinin. AB - Previous studies have demonstrated the importance of the hemagglutinin (HA) of reovirus in determining virus-specific immune responses. In this study, we have used anti-HA monoclonal antibodies and reoviruses with antigenically altered HA proteins to further analyze the CTL response against the HA. We showed that reovirus-specific CTL primarily recognize one distinct antigenic domain on the HA. These results emphasize the fine specificity of the T cell receptor that recognizes viral determinants. The CTL response is directed against the region of the HA that is also responsible for neurotropism and binding to neutralizing antibody. PMID- 6288803 TI - Differential effects of various pharmacological agents on the cytolytic reaction mechanism of the human natural killer lymphocyte: further resolution of programming for lysis and KCIL into discrete stages. PMID- 6288804 TI - Macrophage migration inhibition factor (MIF): reducing the variables. AB - While the phenomenon of macrophage migration inhibition is a useful indicator of lymphokine release, it may be caused by other substances, it is subject to considerable variability, and it may be masked by the concomitant presence of substances stimulating migration. We have investigated certain aspects of the lymphokine macrophage interaction in order to circumvent these problems. Cells from the murine macrophage cell line RAW 264-7 migrated with less variability than fresh guinea pig peritoneal macrophages and were more sensitive target cells for human macrophage migration inhibition factor (MIF). Assays were performed with serum-free and endotoxin-free medium and in all cases in the presence and absence of L-fucose. This added specificity to the assay in that biological MIF activity was invariably blocked by L-fucose whereas migration inhibition produced by antigen complexed antibody, endotoxin, and periodate was not affected by L fucose. It was also possible to demonstrate MIF activity in mixtures of MIF and migration stimulation factor by using L-fucose. We suggest that MIF activity is determined with less variability by using a macrophage cell line as indicator cells and performing the assays in the presence and absence of L-fucose. PMID- 6288805 TI - Nosocomial Legionnaires' disease caused by aerosolized tap water from respiratory devices. AB - Five cases of nosocomial Legionnaires' disease which occurred over a five-month period were retrospectively investigated. Chart review showed that during the two to 10-day incubation period before the onset of illness, all of the patients inhaled aerosolized tap water from jet nebulizers (four patients) or from a portable room humidifier (one patient), and all received high dosages of corticosteroids or adrenocorticotropic hormone. Exposure to both factors was highly significant (P less than 0.000001) when compared with the rate of exposure in 69 control patients. Environmental cultures yielded Legionella pneumophila from tap water and from reservoirs of tap water-filled respiratory devices. The yield was highest from hot tap water, in which the free chlorine level was less than 0.05 parts per million. Thus, Legionnaires' disease may be caused by contaminated aerosols from respiratory devices, and the use of contaminated tap water in such devices represents a previously unrecognized hazard to which corticosteroid-treated patients should not be exposed. PMID- 6288806 TI - The biology and chemistry of Epstein-Barr virus. AB - Epstein-Barr virus (EBV) is the human prototype of a family of closely related herpesviruses of Old World primates. These agents probably evolved and spread among the Old World primates since the divergence of apes from monkeys about 30 million years ago. Although the DNAs of the EBV family have no sequence homology to other herpesviruses, there are some features in common with other herpesviral genomes. EBV DNA is unusual in having five tandem direct repeat elements which divide the genome into five unique sequence domains. The tandem direct repeats at the ends of the linear virion DNA probably mediate circularization of the viral DNA to form the circular episomal viral DNA which is characteristic of EBV infected cells. In latent transforming infection, messenger RNAs are encoded by three widely separate regions of the EBV genome. The remainder of the viral genome encodes many RNAs and proteins which are expressed in productive infection. Early and late viral genes are intermixed along the full length of EBV DNA. PMID- 6288807 TI - Improved test for IgM antibody to Epstein-Barr virus using an absorption step with Staphylococcus aureus. AB - The determination of levels of IgM antibody to Epstein-Barr virus (EBV) capsid antigen was improved by absorbing unfractionated test serum with Staphylococcus aureus cells containing protein A. Nonspecific background fluorescence was significantly decreased, thus facilitating the reading of slides and the detection of specific fluorescent reactions in serum with low levels of this IgM antibody. Moreover, the inclusion of the absorption step eliminated false positive reactions caused by the presence of rheumatoid factor in serum and shortened the time needed for the incubation of test serum with the EBV-infected cell smear. This improved antibody test should facilitate the laboratory diagnosis of acute EBV infections, including unaccompanied by a heterophil antibody response. PMID- 6288808 TI - Potentiation of neutralization of Varicella-Zoster virus to antibody to immunoglobulin. AB - Antibody to varicella-zoster virus was measured in Vero cell cultures by a conventional and an enhanced plaque neutralization test. In the latter, heterologous antibody to human immunoglobulin was added to the virus-serum mixture to potentiate the neutralization reaction. The antibody to human immunoglobulin enhanced the sensitivity of the neutralization test seven- to 107 fold as compared with the conventional procedure. The test was highly reproducible in that individual serum titers fluctuated in a twofold range on repeated testing. A prozone phenomenon was observed in the enhanced neutralization test in low-titered sera, but not with high-titered sera or with specimens of cerebrospinal fluid. The specificity of the test was confirmed by the detection of seroconversion in recipients of live varicella virus vaccine. PMID- 6288810 TI - Poliomyelitis in China. PMID- 6288809 TI - Control of poliomyelitis: a continuing paradox. PMID- 6288812 TI - Practical value of clinical electromyography and nerve conduction studies. PMID- 6288811 TI - Poliomyelitis in the United States, 1969-1981. PMID- 6288813 TI - Effect of murine beta-interferon preparation on phagocytosis and cyclic AMP levels in mouse peritoneal macrophages. AB - Mouse peritoneal macrophages (MPM) cultivated with a mouse beta-interferon preparation (MuIFN-beta) or "mock" IFN were tested for phagocytic ability and intracellular levels of adenosine 3',5'-cyclic monophosphate (cAMP). Suspensions of nonopsonized Escherichia coli (E. coli) were used for phagocytosis experiments. Treatment of MPM with 10(1)-10(3) U per ml of MuIFN-beta stimulated the phagocytic activity and raised the levels of cAMP in MPM. The effect of MuIFN beta on cAMP levels were dose and time dependent. Maximal cAMP levels were seen when MPM were incubated with 10(3)-10(4) U per ml of MuIFN-beta for five hours. Simultaneous addition of MuIFN-beta and the adenylcyclase inhibitor N ethylmaleimide to the MPM cultures prevented the rise in cAMP levels but not the increased phagocytic capacity. MuIFN-beta induced enhancement of phagocytosis and elevation of cAMP levels in MPM seem to be two parallel but not interlinked processes. PMID- 6288814 TI - Human leukocyte-derived alpha-interferons: purification and amino-terminal-amino acid sequence of two components. PMID- 6288815 TI - Interferon production in human hematopoietic cell lines: response to chemicals and characterization of interferons. AB - We have surveyed interferon production and its modulation by 5-bromodeoxyuridine, butyrate, dexamethasone, dimethylsulfoxide and tetradecanoylphorbolacetate in 20 human hematopoietic cell lines derived from leukemias, lymphomas, myelomas and normal leukocytes, representing various maturation stages of lymphoid and myeloid cells. Sendai virus-induced interferon production was enhanced by at least one of the chemicals in 13 out of 14 B-type lymphoid cell lines, whereas no enhancement was observed in any of the non-B, non-T-, T-lymphoid, or myeloid cell lines tested. Interferon produced by 11 cell lines was partially characterized using antisera specific for HuIFN-alpha and HuIFN-beta. Six cell lines produced both IFN-alpha and IFN-beta, two lines produced by IFN-alpha and three lines (including both T-cell lines tested) produced only IFN-beta. In all cases examined, enhancement by chemicals of total interferon yields was due to selective stimulation of production of IFN-alpha. Poly (I):(C) induction of interferon was studied in a number of B-cell lines. In general, a similar pattern of IFN-alpha and -beta synthesis was observed as in virus-induced cells, but the proportion of IFN-beta was relatively smaller. Treatment with butyrate enhanced interferon production to a similar extent accompanied by a similar shift in composition as in virus-induced cells. Our results demonstrate that even in closely related cell types, production of IFN-alpha and IFN-beta can be regulated differently in response to the same inducer. In a single cell type in response to a single inducer, expression of IFN-alpha and IFN-beta can be differentially affected by chemicals. PMID- 6288816 TI - Interferon production in mixed cultures of mouse spleen cells and tumor cells: IFN-gamma as the main component. AB - Interferon (IFN) production in mixed cultures of mouse spleen cells and tumor cells was investigated, mainly using mouse myeloid leukemic cells, M1, and spleen cells from syngeneic SL mice. Spleen cells from other mouse strains (C57BL/6, BALB/c nude and C57BL/6 beige) and other tumor cells (L929 and YAC-1) were similarly effective for IFN production, but mouse mammary carcinoma FM3A cells were not. As the IFN-inducing stimulus, mycoplasma contaminating the tumor cells, or endogenous retrovirus in them, may possibly be involved. The type of IFN produced was identified as IFN-gamma as a major component and IFN-alpha and -beta as minor ones, by acid stability tests and neutralization by antisera with defined specificities against alpha, beta, and gamma. Depletion of natural killer cells from spleen cells did not appreciably affect IFN production, but depletion of Thy 1.2 antigen-bearing cells greatly reduced IFN-gamma production, indicating that T cells were the main IFN producers. PMID- 6288817 TI - Small cell lung cancer at Earl K. Long Memorial Hospital. PMID- 6288818 TI - The intestinal profile of Na-K-ATPase in two rat models of acute renal failure. AB - The specific activity of mucosal Na-K-ATPase in segments of the small intestine and colon was examined after BN or BUL. BN resulted in a surge of the specific activity of the enzyme throughout the mucosa of the intestinal tract (duodenum 26%, jejunum 33%, ileum 37%, and colon 68%). BUL induced to significant change in the specific activity of Na-K-ATPase in the small intestinal mucosa and a small rise (32%) in the specific activity of the enzyme in the colon. Kinetic analyses of potassium activation of the enzyme in the two models of ARF indicate that the increase in the specific activity of the enzyme in the colonic mucosa was a function of increased Vmax rather than a change in the apparent Km for potassium by the Na-K-ATPase. It is theorized that the kidney modifies the response of intestinal Na-K-ATPase in AFR. PMID- 6288819 TI - Beta-thalassemia with exceptionally high hemoglobin A2. Differential expression of the delta-globin gene in the presence of beta-thalassemia. AB - Hemoglobin A2 levels in normal adults are rarely greater than 3.5%. In patients heterozygous for beta-thalassemia, they average about 5% but do not usually exceed 7%. We studied a family in which four patients with heterozygous beta thalassemia had HbA2 levels of 8.4% to 11.2%. Globin biosynthesis studies and restriction endonuclease mapping of the alpha-globin loci showed homozygous or heterozygous alpha-thalassemia-2 as well as beta-thalassemia in some family members. The delta- and beta-globin genes were examined by using the restriction enzymes Eco RI, Pvu II, and Xba I, which cut both within and outside the coding portions of the delta- and beta-loci. Only the expected delta- and beta-globin gene containing fragments were present, excluding a crossover event producing a fusion gene that would code for delta-globin but possibly be under the regulatory influence of nucleotide sequences that control the expression of the beta-gene. This kindred provides evidence that in the presence of beta-thalassemia, expression of the delta-gene, beyond that commonly seen, is possible. This could be a direct result of the gene defect producing beta-thalassemia or be due to differences in the delta-globin gene linked to this beta-thalassemia gene. The interactions of alpha- and beta-thalassemia may alter tetramer assembly and increase HbA2 levels; however, this possibility seems less likely. PMID- 6288820 TI - The transport and accumulation of oxyvanadium compounds in human erythrocytes in vitro. AB - Metavanadate, at physiologic pH the oxyanion form of pentavalent vanadium, is a potent reversible inhibitor of the sodium pump. Vanadium must enter cells to inhibit the sodium pump, and metavanadate may be converted to an inactive form inside of cells. Because of these factors and the complex inorganic chemistry of vanadium, we examined the kinetics of vanadium uptake and accumulation in normal erythrocytes in vitro at 37 degrees C. The kinetics of vanadium influx, efflux, and accumulation in erythrocytes in Tris-buffered, isotonic salt medium were fit closely by a model with vanadium in two possible oxidation states and with the vanadium permeating between two compartments. The equation for this model is: (formula: see text) in which subscripts i and o signify "inside" and "outside" the cells, respectively, and k1, k-1, and k2 are rate constants. 48V or EPR of vanadium(IV) gave similar estimates of the concentrations of the components. The k1 was 0.37 +/- 0.06 (S.E.M.) min-1 and k2 was 0.04 +/- 0.01 min-1. In control Tris-medium, k1 exceeded k-1 by a factor of 1.8. After 120 min of incubation in media with initial concentrations of 1, 10, or 100 microM vanadium(V), the total intracellular vanadium concentration exceeded that in the bath 4.5 to 18-fold. Vanadium influx was not appreciably changed by variations of external sodium or glucose levels. The k1 and k-1 were inversely related to external pH over the range 6.5 to 8.2. High O2 tension (95% to 100% O2) caused a decrease in k2, and the lipophilic oxidant, cumene hydroperoxide, accelerated the loss of accumulated vanadium from the cells, indicating that the k2 step represents reduction of vanadium(V) to vanadium(V) within the cells. On the basis of these studies we suggest that the intracellular concentration of vanadium(V), the inhibitor of the sodium pump, is determined by the combined effects of the rate of vanadium influx (dependent on the extracellular concentration of free metavanadate), the rate of vanadium efflux, and the rate of conversion of vanadium(V) to vanadium(IV). PMID- 6288821 TI - Myoepithelioma of the external auditory canal. PMID- 6288823 TI - In vivo study of interference between herpes and influenza viruses. PMID- 6288822 TI - HMG-CoA reductase kinase: measurement of activity by methods that preclude interference by inhibitors of HMG-CoA reductase activity or by mevalonate kinase. AB - Assay of HMG-CoA reductase kinase activity requires HMG-CoA reductase (reductase, E.C. 1.1.1.34) free of associated reductase kinase. Microsomal reductase insensitive to inactivation by Mg-nucleotides alone may be prepared by heating microsomes at 50 degrees C for 15 min. The reductase in these microsomes may subsequently be inactivated by Mg-nucleotides only after addition of reductase kinase. Inactivation is a linear function of time and of cytosol protein concentration and may be reversed by treatment with a phosphoprotein phosphatase. The extent of inactivation observed under standard conditions provides an assay for reductase kinase activity. Factors present in cytosol that hinder measurement of either reductase or reductase kinase activity must be removed or inhibited. Reductase phosphatase is inhibited by 50 mM NaF. Reductase kinase kinase activity is not expressed under the assay conditions used. Mg-Nucleotide-independent inhibitors of reductase activity are removed by chromatography on DEAE-Sephacel or Blue Sepharose. Mevalonate kinase and reductase kinase are separable by chromatography on DEAE-Sephacel or Sephadex G-200. We describe a rapid chromatographic procedure for separating reductase kinase of crude fractions from mevalonate kinase and from Mg-nucleotide-independent inhibitors of reductase activity. The 1.0 M KCl eluate from DEAE-Sephacel contains all of the cytosol reductase kinase activity. This method is applicable to measurement of reductase kinase activity in cytosol or more purified fractions. PMID- 6288824 TI - Study of platelet aggregation in vivo. IX. Effect of nafazatrom on in vivo platelet aggregation and spontaneous tumor metastasis. AB - Nafazatrom (Bay g 6575) was explored for its ability to inhibit platelet aggregation. In vitro, it had no effect on ADP, serotonin, epinephrine, or collagen induced platelet aggregation in platelet rich plasma of monkeys. On the other hand, in vivo it was a powerful inhibitor of ADP induced platelet aggregation as measured by the in vivo platelet aggregation recording instrument described previously (Ambrus et al., 1976). This effect was potentiated by dipyridamole. On the other hand, following parenteral administration of Bay g 6575, no ex vivo inhibition was noticed of ADP, serotonin, epinephrine, and collagen induced platelet aggregation. The hypothesis was presented that Bay g 6575 acts by increasing prostacyclin synthesis and/or release or interferes with its decomposition. This may explain in vivo activity; rapid decomposition may explain inability to demonstrate ex vivo activity. This also explains potentiation by the phosphodiesterase inhibitor dipyridamole. Bay g 6575 also was highly effective as a platelet aggregation inhibitor in monkeys after oral administration. In mice, Bay g 6575 increased circulation time of intravenously injected polyploid Ehrlich ascites tumor cells. In Furth-Wistar rats implanted with Furth-Columbia Wilms' tumor, in A/J mice implanted with C1300 neuroblastoma and in Wistar rats implanted with SMT-2A (Kim) breast cancer, Bay g 6575 significantly reduced spontaneous pulmonary metastasis. On the other hand, no effect was seen in the metastatic rate of NIH renal adenocarcinoma in BALB/cCr mice. PMID- 6288826 TI - Small amplitude periodic waves for the FitzHugh-Nagumo equations. PMID- 6288825 TI - Failure of low temperature to arrest degradation of 14C-adenosine 5'-diphosphate by human plasma in the presence of EDTA. AB - Degradation of 14C-ADP in human platelet-rich and platelet-poor plasmas was studied. Three procedures were tested to determine which method was most effective in "stopping" enzyme reactions. One involved the immediate addition of perchloric acid to the reaction mixture at the end of the incubation period. Another involved "stopping" enzyme reactions at 0 degree C for about 20 min before the addition of perchloric acid. The third involved adding EDTA (10 mM final concentration) to "stop" the reactions at the end of incubation. The protein-free extracts from the plasmas were processed for adenine nucleotide assays by thin-layer chromatography. Addition of perchoric acid was most effective. Low temperature did not stop degradation of 14C-ADP in plasma even in the presence of EDTA. 14C-ADP was converted primarily to 14C-AMP. ADPase activity was the same in platelet-rich and platelet-poor plasmas. We conclude that the addition of EDTA and the cooling of plasma did not completely stop the enzymic degradation of ADP. PMID- 6288827 TI - Long-term survival in small cell carcinoma of the lung with associated symptomatic intracranial metastases. PMID- 6288828 TI - Cefotaxime in the treatment of gonorrhoea caused by penicillinase producing N. gonorrhoeae (PPNG) and non penicillinase producing N. gonorrhoeae (Non PPNG). PMID- 6288829 TI - Cyclic GMP concentration in ovarian tissue after administration of gonadotrophin to prepubertal rats. AB - The weights, protein concentrations, and concentrations of cyclic AMP and cyclic GMP in the ovaries of prepubertal rats have been measured after induction of follicular growth by administration of pregnant mare serum gonadotrophin (PMSG). Three phases of ovarian growth, the follicular growth phase, the preovulatory phase and the luteal phase were investigated. Ovulation was induced by administration of human chorionic gonadotrophin (hCG). During the follicular phase, ovarian weight, protein content and the content of both cyclic AMP and cyclic GMP increased 2-3 days after administration of PMSG. Administration of hCG 65h after PMSG caused a rapid rise in the concentration of cyclic AMP but the concentration of cyclic GMP tended to fall. During the luteal phase the concentrations of both cyclic nucleotides decreased. PMID- 6288830 TI - Cell membrane regionalization in early mouse embryos as demonstrated by 5' nucleotidase activity. AB - The distribution of 5'-nucleotidase activity in pre-implantation mouse embryos is studied by means of a cytochemical method adapted from Uusitalo & Karnovsky (1977). The enzyme activity is detected, from the 4-cell stage up to the morula stage, on discrete patches of the cell membrane between blastomeres. Appropriate controls show that this distribution is not a localization artifact due to selective retention of the enzyme reaction product in the narrow interblastomeric spaces. In early blastocysts, as the blastocysts expands the enzyme activity on its lining disappears. The external surface of the trophectoderm in early blastocysts lacks any enzyme activity, whereas in late blastocysts a strong enzyme activity is detected at the embryonic trophectoderm, decreasing in intensity towards the opposite pole of the embryo. These results are compared to previous observations by other authors and the differences are mainly ascribed to differences in the cytochemical procedure employed. We conclude that during cleavage a gradual cell membrane regionalization unfolds, revealing a pattern that may be related to morphogenesis; in particular, to the localization of zonular tight junctions around the peripheral blastomeres of the morula (Izquierdo, 1977; Izquierdo, Lopez & Marticorena, 1980). PMID- 6288831 TI - Studies on antigens useful for detection of IgE antibodies in isocyanate sensitized workers. AB - Toluene diisocyanate was conjugated by two different methods to human serum albumin. These conjugates were used as antigens in the radio-allergo-sorbent-test (RAST) for detection of IgE antibodies in sera of isocyanate-sensitized persons. Chemical analyses have shown that conjugates containing 18-28 mol of isocyanate residues per mol protein were more potent antigens than those containing greater amounts of isocyanate residues. These results were confirmed by RAST inhibition experiments. The importance of optimization of the immunoradiometric methods for study of immunologic sensitization to isocyanates is discussed. PMID- 6288832 TI - Objectifying psychiatric diagnosis and treatment with the dexamethasone suppression test. AB - The application of the dexamethasone suppression test (DST) to the treatment of depressive illness is discussed from the perspectives of its use in diagnosis, the measurement of outcome, and the prediction of response to antidepressant medications. Patients with nonsuppressing depression occur in about one half of the endogenously depressed population and probably represent a norepinephrine deficient subtype of the syndrome. There is clear evidence that a change in the DST from nonsuppression to suppression parallels recovery from a depressive illness, and this may be used as an objective measure of outcome. Early studies show promise that the DST can predict the antidepressant of choice, although there are some limitations of the DST in treatment choice. PMID- 6288834 TI - [Serological studies in early stage of nasopharyngeal carcinoma ]. PMID- 6288833 TI - Suppression of male pronuclear movement in frog eggs by hydrostatic pressure and deuterium oxide yields androgenetic haploids. AB - The disruptive effects of microtubule-specific agents on pronuclear movement illustrate the requirement of an intact cytoskeletal system for movement. In this study, we investigated the effects of high hydrostatic pressure and deuterium oxide (D2O) on fertilized Rana pipiens eggs during the time of pronuclear migration. The eggs were either pulsed for six min with 3000, 5000, or 7000 psi or placed for ten min in 80% D2O between the time of second polar body emission and first cleavage. Both treatments disrupted male pronuclear migration as shown by eccentric first cleavage furrows. Treatment of eggs prior to pronuclear association resulted in haploid production. The androgenetic origin of the haploid embryos was demonstrated using morphological and isozymal markers produced by the cross Rana pipiens female x Rana utricularia male. Eggs treated with D2O also yielded embryos with neural defects identical to those following ultraviolet irradiation. This study complements the recent reports on pressure suppression of the second polar body and of first cleavage by showing that the selective suppression of microtubular function between these two events produces an entirely different set of genetic and developmental consequences. PMID- 6288835 TI - Aminopyridine block of transient potassium current. AB - The blocking action of 4-aminopyridine (4-AP) and 3, 4-diaminopyridine (Di-AP) on transient potassium current (IA) in molluscan central neurons was studied in internal perfusion voltage-clamp experiments. Identical blocking effects were seen when the drugs were applied either externally or internally. It was found that aminopyridines have two kinds of effects on IA channels. The first involves block of open channels during depolarizing pulses and results in a shortening of the time to peak current and an increase in the initial rate of decay of current. This effect of the drug is similar to the block of delayed potassium current by tetraethylammonium (TEA). The other effect is a steady block that increases in strength during hyperpolarization, is removed by depolarization, and is dependent on the frequency of stimulation. The voltage dependence of steady state block approximates the voltage dependence of inactivation gating a changes e-fold in approximately 10 mV. These data suggest that the strength of block may depend on the state of IA gating such that the resting state of the channel with open inactivation gate is more susceptible to block than are the open or inactivated states. A multistate sequential model for IA gating and voltage-dependent AP block is developed. PMID- 6288836 TI - Physiological evidence that light-mediated decrease in cyclic GMP is an intermediary process in retinal rod transduction. AB - Brief, intracellularly injected pulses of cyclic GMP transiently depolarized toad retinal rod outer segments (ROS). The depolarization is antagonized by light, perhaps by the activation of phosphodiesterase (PDE), as shown in the biochemical studies of others. As measured by the antagonism of cyclic GMP pulses by light, PDE activity peaks after the peak of the receptor potential and has approximately the same recovery time as the membrane voltage after weak illumination, but recovers more slowly than the membrane potential after strong illumination, as sensitivity does in other preparations. A cyclic GMP pulse delivered just after the hyperpolarizing phase of the receptor potential tends to turn off the light response. The kinetics of recovery from this turnoff are similar to those of the initial phase of the receptor potential. This similarity suggests that the initial phase of the receptor potential is controlled by light-activated PDE. Both EGTA and saturating doses of cyclic GMP block the light response, but only cyclic GMP increases response latency, which suggests that if calcium is involved in transduction, it is controlled by the hydrolysis of cyclic GMP. After brief pulses of cyclic AMP, a new steady state of increased depolarization occasionally develops. The effects described above also occur under these conditions. The results are consistent with the hypothesis that light-activated hydrolysis of cGMP is an intermediary process in transduction. PMID- 6288837 TI - Kinetics of oxygen consumption after a single flash of light in photoreceptors of the drone (Apis mellifera). AB - The time course of the rate of oxygen consumption (QO2) after a single flash of light has been measured in 300-micrometers slices of drone retina at 22 degrees C. To measure delta QO2(t), the change in QO2 from its level in darkness, the transients of the partial pressure of O2 (PO2) were recorded with O2 microelectrodes simultaneously in two sites in the slice and delta QO2 was calculated by a computer using Fourier transforms. After a 40-ms flash of intense light, delta QO2, reached a peak of 40 microliters O2/g.min and then declined exponentially to the baseline with a time constant tau 1 = 4.96 +/- 0.49 s (SD, n = 10). The rising phase was characterized by a time constant tau 2 = 1.90 +/- 0.35 s (SD, n = 10). The peak amplitude of delta QO2 increased linearly with the log of the light intensity. Replacement of Na+ by choline, known to decrease greatly the light-induced transmembrane current, caused a 63% decrease of delta QO2. With these changes, however, the kinetics of delta QO2 (t) were unchanged. This suggest that the recovery phase is rate-limited by a single reaction with apparent first-order kinetics. Evidence is provided that suggests that this reaction may be the working of the sodium pump. Exposure of the retina to high concentrations of ouabain or strophanthidin (inhibitors of the sodium pump) reduced the peak amplitude of delta QO2 by approximately 80% and increased tau 1. The increase of tau 1 was an exponential function of the time of exposure to the cardioactive steroids. Hence, it seems likely that the greatest part of delta QO2 is used for the working of the pump, whose activity is the mechanism underlying the rate constant of the descending limb of delta QO2 (t). PMID- 6288839 TI - Purification of Chlamydia trachomatis lymphogranuloma venereum elementary bodies and their interaction with HeLa cells. AB - A procedure has been developed to yield infectious elementary bodies of the lymphogranuloma venereum strains LGV 434 and 404 of Chlamydia trachomatis, labelled during intracellular growth in HeLa 229 cells. The final preparation, obtained after velocity sedimentation of a polycarbonate membrane-filtered sample through a sucrose gradient, is free of host proteins and, more importantly, of chlamydial reticulate bodies. Using such purified preparations, it was found that the association of LGV 434 elementary bodies with HeLa 229 cultures was unaffected by the pretreatment of the host cells with a variety of lectins or with neuraminidases from Clostridium perfringens and Vibrio cholerae. The association was inhibited by dextran sulphate and by mild trypsin treatment of HeLa cultures. Treatment of purified elementary bodies with trypsin, chymotrypsin, neuraminidases and a variety of carbohydrates and lectins did not produce any change in the rate of association with HeLa cultures. Heat inactivated elementary bodies were significantly less able to associate with the host cells. PMID- 6288840 TI - Spontaneous deletions in the TOL plasmid pWW20 which give rise to the B3 regulatory mutants of Pseudomonas putida MT20. AB - The size of the TOL plasmid pWW20 from Pseudomonas putida MT20, as measured by analysis of agarose electrophoresis gels after restriction endonuclease hydrolysis, was 270-280 kilobase pairs (kb). During growth on benzoate, MT20 segregates strains carrying mutations in the plasmid regulatory gene xylS; these so-called B3 strains retain the ability to grow on m-xylene (Mxy+) but do not grow on its metabolite m-toluate (Mtol-) and have also lost the ability to transfer the plasmid (Tra-). Analysis of restriction digests of plasmid DNA from seven such segregants, independently isolated, showed that pWW20 had undergone extensive deletions of 90-100 kb. All the deleted plasmids had lost a common core of DNA, of about 72-80 kb, but in class A mutants the deletion extended at one end of this core and in class B mutants at the other end. Class A and B mutants also differed in their rate of growth on m-xylene as a result of differences in the level of expression of their plasmid-coded catabolic enzymes. This suggests that an additional gene, involved in regulating levels of gene expression, is located in the region uniquely deleted in the class B mutants. PMID- 6288838 TI - Delays in inactivation development and activation kinetics in myxicola giant axons. AB - Na inactivation was studied in Myxicola (two-pulse procedure, 6-ms gap between conditioning and test pulses). Inactivation developed with an initial delay (range 130-817 microseconds) followed by a simple exponential decline (time constant tau c). Delays (deviations from a simple exponential) are seen only for brief conditioning pulses were gNa is slightly activated. Hodgkin-Huxley kinetics with series resistance, Rs, predict deviations from a simple exponential only for conditioning pulses that substantially activate gNa. Reducing INa fivefold (Tris substitution) had no effect on either tau c or delay. Delay in not generated by Rs or by contamination from activation development. The slowest time constant in Na tails is approximately 1 ms (Goldman and Hahin, 1978) and the gap was 6 ms. Shortening the gap to 2 ms had no effect on either tau c or delay. Delay is a true property of the channel. Delay decreased with more positive conditioning potentials, and also decreased approximately proportionally with time to peak gNa during the conditioning pulse, as expected for sequentially coupled activation and inactivation. In a few cases the difference between Na current values for brief conditioning pulses and the tau c exponential could be measured. Difference values decayed exponentially with time constant tau m. The inactivation time course is described by a model that assumes a process with the kinetics of gNa activation as a precursor to inactivation. PMID- 6288841 TI - Evidence for the presence of two terminal oxidases in the trypanosomatid Crithidia oncopelti. AB - Increasing concentrations of cyanide inhibited the respiration of whole cells of Crithidia oncopelti in a biphasic fashion. Approximately 80% inhibition was attained with 40 microM-KCN. No further inhibition occurred until the concentration of KCN reached approximately 200 microM. Thereafter inhibition rose gradually to 100% at 1500 microM-KCN. Difference spectra revealed the presence of two CO-reacting haemoproteins. These were shown to be two different functional oxidases by photochemical action spectra obtained by using laser light. One oxidase was identified as cytochrome a + a3 whilst the other had the properties of cytochrome o. Both oxidases could be detected in cells at all stages of growth by the above methods. PMID- 6288842 TI - Bacillus subtilis extracellular nuclease production associated with the spoOH sporulation locus. AB - A number of nuclease-deficient mutants of Bacillus subtilis were isolated and found to be concurrently asporogenous. The nuclease-deficient phenotype appeared to be associated with the spoOH sporulation locus. Spontaneously occurring sporogenous revertants concomitantly recovered the ability to produce extracellular nuclease activity. The position of the ncl mutation was determined using transformation and PBS1 transduction and found to map in the same site as spoOH. The map order of ncl and markers in the vicinity was determined to be purA cysA-ncl(spoOH)-strA. PMID- 6288843 TI - The maintenance and propagation of plasmid genes in bacterial populations. The Sixth Fleming Lecture. PMID- 6288844 TI - Cyclic AMP and cyclic GMP control of synthesis of constitutive enzymes in Escherichia coli. AB - Escherichia coli was grown in chemostat culture under glycerol-limited and ammonium-limited conditions at growth rates between 0.1 and 0.5 h-1. At steady state, the concentrations of cyclic AMP and cyclic GMP and the activities of four constitutive enzymes (glucose-6-phosphate dehydrogenase, isocitrate dehydrogenase, NADH oxidase and cyclic phosphodiesterase) were determined in the organism. Addition of exogenous cyclic AMP, cyclic GMP or phencyclidine perturbed the steady state and caused inhibition or stimulation of synthesis of phosphodiesterase and isocitrate dehydrogenase. A novel hypothesis is proposed to account for the ability of bacteria to regulate the synthesis of constitutive enzymes with cyclic nucleotides and possibly other small molecules. PMID- 6288845 TI - Evidence for non-chromosomal hepatitis B virus surface (HBsAg)- and core antigen (HBcAg)-specific DNA sequences in a hepatoma cell line. AB - As demonstrated previously, a "beta particle" fraction isolated from the cytoplasm of PLC/PRF/5 cells contains hepatitis B virus (HBV)-specific DNA. Here, further evidence is provided that the specificity of the DNA for HBV is represented at least by sequences coding for the surface and core antigen (HBsAg and HBcAg). This was shown by two different hybridization techniques. One of them, the technique of Southern, distinguished these hybrid molecules formed from those containing HBV DNA integrated into chromosomes. The HBV-specific beta particle DNA forms two distinct bands separate from the high molecular weight cellular DNA. PMID- 6288846 TI - Failure of guanidine and 2-(alpha-hydroxybenzyl)benzimidazole to inhibit replication of hepatitis A virus in vitro. AB - Replication of hepatitis A virus (HAV) in the human hepatoma-derived PLC/PRF/5 cell line was neither inhibited in the presence of various concentrations of guanidine or D-2-(alpha-hydroxybenzyl)benzimidazole (D-HBB), nor were the two chemicals effective in combination. Under identical conditions, however, replication of poliovirus type 1 was inhibited. Tracer experiments with radiolabelled guanidine and D-HBB also furnished no evidence that the two antiviral substances were metabolized gradually to inactive derivatives in PLC/PRF/5 cells. Therefore, it is concluded that resistance to the action of guanidine and D-HBB is an inherent characteristic of HAV. However, the insensitivity of HAV to these drugs does not exclude the virus from the family of picornaviruses. PMID- 6288847 TI - Early and delayed shut-off of host protein synthesis in cells infected with herpes simplex virus. AB - A mutant of herpes simplex virus type 1 [HSV-1(HFEM)], tsB7, appears to have two temperature-sensitive functions. One is required during the first hour of infecting a cell (suggesting that it is performed by a virion protein) and the other is the non-essential function of "early shut-off" of cellular protein synthesis, which is mediated by a virion protein. The latter function remained temperature-sensitive in a revertant virus (RC2) grown at the non-permissive temperature (39 degrees C). However, under these conditions RC2 did cause inhibition of host synthesis, showing that "delayed shut-off", requiring virus protein synthesis, can occur independently of early shut-off, which is mediated by a virion protein. Early shut-off by u.v.-irradiated tsB7 was reversed when the temperature was raised, whereas delayed shut-off by intact tsB7 was not. Of two wild-type strains of virus examined, HSV-1(F) also exhibited temperature sensitive early shut-off, but HSV-2(G) did not. PMID- 6288849 TI - Effect of Nu/Nu gene on genetically determined resistance to murine cytomegalovirus. AB - Adult athymic Nu/Nu mice showed increased susceptibility to lethal infection with murine cytomegalovirus (MCMV) when compared to their heterozygous T cell competent Nu/+ littermates. However, the extent of this increase in susceptibility varied dramatically depending on the genetic background of the mice carrying the Nu/Nu gene. Genetically susceptible Balb/c (H-2d) mice showed a greater than 316-fold difference between the LD50 of Nu/Nu and Nu/+ littermates. In marked contrast, the genetically resistant CBA (H-2K) strain was characterized by only a 16-fold difference in resistance between Nu/Nu and Nu/+ mice, and furthermore, the athymic CBA Nu/Nu mice were no susceptible than the T cell competent Balb/c Nu/+ strain. These results together with previous observations strongly suggest that the (H-2k)-associated resistance of the CBA strain is mediated by non-T cell-dependent early defence mechanisms. PMID- 6288848 TI - Pathogenesis of herpes simplex virus in B cell-suppressed mice: the relative roles of cell-mediated and humoral immunity. AB - B cell responses of Balb/c mice were suppressed using sheep anti-mouse IgM serum. At 4 weeks, both B cell-suppressed and normal littermates were infected in the ear pinna with herpes simplex virus type 1 (HSV-1). The B cell-suppressed mice failed to produce neutralizing herpes antibodies in their sera but had a normal cell-mediated immunity (CMI) response as measured by a delayed hypersensitivity skin test. Although the infection was eliminated from the ear in both B cell suppressed and normal mice by day 10 after infection, there was an indication that B cell-suppressed mice had a more florid primary infection of the peripheral and central nervous system and also a higher incidence of a latent infection. These results support the hypothesis that antibody is important in restricting the spread of virus to the central nervous system, whereas CMI is important in clearing the primary infection in the ear pinna. PMID- 6288850 TI - Protein synthesis in HeLa cells double-infected with encephalomyocarditis virus and poliovirus. PMID- 6288851 TI - Comparative physicochemical and biological properties of two strains of Kilham rat virus, a non-defective parvovirus. AB - Two antigenically indistinguishable strains, 171 and 308, of Kilham rat virus (KRV) have distinct host ranges and contain capsid proteins of identical size, but with different isoelectric points. The single-stranded DNA genomes of the viruses are also the same size but appear to have different secondary and tertiary structures. The genomes of the two strains have nearly identical cleavage maps of 11 restriction endonucleases, except for differences in five restriction sites in the region bracketed by 0.63 to 0.90 map units (from the 3' ends of the virus strands). However, there is a lack of extended heterology in the nucleotide sequence of the two virus genomes, as judged by electron microscopic analysis of the heteroduplex of the two virus DNAs. This suggests that very subtle differences in the sequences of the genome, and possibly of the capsid proteins, may play a role in the host specificity without affecting the antigenic similarity of KRV strains. PMID- 6288852 TI - Murine xenotropic type C viruses. IV. Replication and pathogenesis of ducks. AB - The xenotropic (X-tropic) mouse type C virus (MuLV) and its pseudotype of murine sarcoma virus (MSV) were inoculated into several fertilized developing Pekin duck eggs. The development of the duck embryos was substantially reduced in those receiving the X-tropic viruses compared to eggs inoculated only with tissue culture medium. Infections virus was isolated from some of the adult animals; in others, evidence for integrated virus sequences in the tissues was noted. No specific pathology was found in the ducks that received X-trophic MuLV alone, but one duck developed multiple fibrosarcomas when inoculated at birth with the X tropic virus pseudotype of MSV. Two ducks receiving X-tropic MuLv had signs of haematopoietic disorders. In addition, more virus-inoculated animals had evidence of hepatitis and encephalitis than control ducks. Antibody production to X-tropic MuLv was present in several ducks inoculated with virus either in embryo or at birth. Absence of antiviral antibodies was noted in those animals whose tissue contained replicating virus. These studies confirm the observations with X-tropic virus in tissue culture. They demonstrate in vivo that avian species are susceptible to infection by the mouse X-tropic virus and that their fibroblasts can be transformed by the X-tropic MuLV pseudotype of MSV. PMID- 6288853 TI - Fusion injection of Rous Sarcoma virus proteins into Rous sarcoma virus transformed, non-producing hamster cells causes release of infectious virus. AB - Purified virus proteins from transformation-defective (td) mutants of Rous sarcoma virus PrA or PrB were trapped in human erythrocyte ghosts which, after resealing, were fusion-injected into hamster RBH cells or rat TWERC cells. These cell lines are non-productively transformed by subgroup C Rous sarcoma virus. After fusion injection the hamster RBH cells released transforming subgroup C Rous sarcoma virus. No infectious virus could be rescued from rat TWERC cells. Since previous experiments have shown that fusion injection of the purified Rous sarcoma virus protein p15 into hamster RBH cells caused cleavage of the precursor protein pr76 to form the virus group-specific antigen (gag) but did not induce infectious virus, we conclude that in addition to p15 other virus proteins are required to induce virus rescue in hamster RBH cells. PMID- 6288854 TI - Characterization of Theiler's murine encephalomyelitis virus RNA. AB - Theiler's murine encephalomyelitis viruses are usually included in the enterovirus genus of the family Picornaviridae, although there is little physicochemical evidence to support this classification. In this report, the size of the RNA of highly virulent and less virulent representatives of the Theiler's group of viruses has been determined by sucrose gradient centrifugation and electrophoresis in agarose to be the same as that of other enteroviruses. The absence of a poly(C) residue provides evidence that these viruses are not cardioviruses or aphthoviruses. The base composition of the two members are similar to each other but differ from those of other enteroviruses. However the one- and two-dimensional maps of the ribonuclease T1 hydrolysates of the two virus RNAs show considerable differences despite their close serological similarity. Virus-specified RNA synthesis in cells infected with the more virulent strain of the virus was almost 10 times greater than that induced by the less virulent strain, in accord with the yields of virus particles. PMID- 6288855 TI - Development of protease activation mutants of HVJ (Sendai virus) in persistently infected cell cultures. AB - HVJ wild-type virus, in which the F protein is activated by trypsin but not by elastase, was spontaneously converted to a mutant with an F protein characterized by being activated by elastase alone. This spontaneous mutation generally occurred during serial passages of cells persistently infected with HVJ, even though the cells were first established by infection with plaque-purified wild type virus. Multiple-cycle replication, plaque formation, haemolysis and SDS polyacrylamide gel electrophoretic (SDS-PAGE) analysis showed that all the elastase-activated mutants isolated from HVJ carrier cells no longer required trypsin for F protein activation. At early passages, these protease activation mutants did not show temperature-sensitive (ts) growth, while at a later stage the mutants, together with the ts mutation, appeared dominant. The frequency of such a protease activation mutation during passage in the HVJ carrier cells seemed to depend on the cell species, but was increased when compared to lytic infections. PMID- 6288856 TI - The isolation and characterization of mutants of herpes simplex virus type 1 that induce cell fusion. AB - Six cell fusion-causing syn mutants were isolated from the KOS (syn-101 to syn 106) and three from the HFEM (syn-109) strains of herpes simplex virus type 1 (HSV-1). The mutants were studied by complementation and recombination with syn 20 (a syncytial mutant of KOS) and ts-B5 (a syncytial mutant of HFEM). Some studies also employed MP, a syncytium-inducing strain isolated from the non syncytial parent, mP. Complementation and recombination of syn-20 and ts-B5 indicated that these two mutants were altered in two different virus genes. The recombination frequency between syn-20 and ts-B5 was very similar to that observed between MP and ts-B5, indicating the syn-20 and MP may represent alterations in the same virus gene. syn-101, syn-103, syn-104 and syn-105 were tentatively assigned to the syn-20 complementation group, while syn-107 and syn 109 were tentatively assigned to the ts-B5 complementation group, syn-106 and syn 108 were excluded from the ts-B5 group. syn-102 could not be excluded from either complementation group. syn-101 induced markedly less fusion at 38 degrees C relative to 34 degrees C. At 34 degrees C the patterns of syn-101-infected cell peptides and glycopeptides, examined by SDS-gel electrophoresis, were normal, but at 38 degrees C the amount of glycopeptide gC was particularly reduced. syn-102 produced decreased amounts of glycoproteins, and a non-glycosylated peptide, probably ICP6, was absent from extracts infected with syn-106. PMID- 6288857 TI - Polypeptides of varicella-zoster virus (VZV) and immunological relationship of VZV and herpes simplex virus (HSV). AB - Varicella-zoster virus (VZV), labelled with [35S]methionine or [14C]glucosamine, was purified by centrifugation through sucrose gradients followed by equilibrium centrifugation in CsCl, and analysed by SDS-polyacrylamide gel electrophoresis (SDS-PAGE). At least 32 polypeptides ranging in mol. wt. from approx. 280 X 10(3) to 21.5 X 10(3) (280K to 21.5K) and six glycopeptides ranging in mol. wt. from approx. 115K to 45K (gp1 to gp6) were found in the virion. The immunological relationship of VZV and herpes simplex virus (HSV) was investigated. In neutralization (NT) tests, no cross-neutralization was observed between VZV and HSV-1 or -2. In fluorescent antibody staining, however, a cross-reaction was observed between VZV- and HSV-1-infected human embryonic lung (HEL) cells and heterologous antiserum. When cross-reactions were investigated by immunoprecipitation followed by SDS-PAGE, several cross-reacting polypeptides were discovered. Cross-reacting glycopeptides of 64K (gp3) and 55K (gp5) were isolated from VZV-infected cell lysates by affinity column chromatography to immobilized HSV-1 antibodies. PMID- 6288858 TI - Virus particles and glycoprotein excreted from cultured cells infected with varicella-zoster virus (VZV). AB - Virus particles and virus proteins excreted from cultured human embryonic lung (HEL) cells infected with varicella-zoster virus (VZV) were examined by electron microscopy and affinity column chromatography using an antibody to VZV followed by SDS-polyacrylamide gel electrophoresis (SDS-PAGE). Approximately 1 X 10(9) to 2 X 10(9) virus particles/ml with no detectable infectivity, of which 30 to 80% were enveloped, were observed in the culture fluid 48 to 72 h after infection, when cytopathic effect (c.p.e.) appeared. In the sonicated infected cell suspension, 1 X 10(9) to 2 X 10(9) virus particles/ml, of which 30 to 50% were enveloped, were observed and the virus particle/infectivity ratio was approx. 10(6):1. The culture fluid of infected HEL cells labelled with [35S]methionine or [3H]glucosamine was centrifuged at 1000000 g for 2 h to remove virus particles and the supernatant was examined for excreted virus proteins. Affinity column chromatography of the supernatant using immobilized human zoster convalescent serum, led to the isolation of virus antigens which were analyzed by SDS-PAGE. Polypeptides with mol. wt. of approx. 115K and 45K, both of which were glycosylated, were detected, suggesting that these VZV glycoproteins were excreted from the infected cells. PMID- 6288859 TI - Neurochemical studies on the cerebellar hypoplasia of Gunn rat (hereditary hyperbilirubinemic rat). AB - The cerebellar hypoplasia induced by hereditary hyperbilirubinemia in the Gunn rat was analyzed neurochemically and immunohistochemically. The antiserum against myelin basic protein was used to visualize the arborization of the fibers in the cerebellum. Arborization was very scarce in the affected lobes of the homozygous (jj) cerebellum. Na,K-ATPase activity did not show significant differences between the jj and the control (Jj) cerebellum. The concentration of norepinephrine in the jj cerebellum was about 1.5 times that of the control. However, the activation ratio of the Na,K-ATPase by norepinephrine and other catecholamines such as dopamine and isoproterenol was about twice as high as the basal activity, and no significant difference was observed between the jj and the Jj cerebella. The glutamic acid decarboxylase activity of the jj cerebellum did not differ significantly from that of the control. PMID- 6288860 TI - Cerebellar hypoplasia in Gunn rats: effects of bilirubin on the maturation of glutamate decarboxylase, Na,K-ATPase, 2',3'-cyclic nucleotide 3' phosphohydrolase, acetylcholine and aryl esterase, succinate and lactate dehydrogenase, and arylsulfatase activities. PMID- 6288861 TI - Inhibition of [3H]diazepam and [3H]3-carboethoxy-beta-carboline binding by irazepine: evidence for multiple "domains" of the benzodiazepine receptor. AB - The binding of [3H]diazepam and [3H]3-carboethoxy-beta-carboline was examined in rat brain synaptosomal membranes treated with irazepine, an alkylating benzodiazepine. Under incubation conditions that resulted in a 25-33% reduction in the Bmax of [3H]diazepam binding, only modest (less than 8.5%) reductions in the Bmax of [3H]3-carboethoxy-beta-carboline were observed. The differential effects of irazepine on the binding of these two compounds may be explained by the presence of multiple areas or "domains" on the benzodiazepine receptor. PMID- 6288862 TI - Regulation of cyclic AMP accumulation in a rat sympathetic ganglion: effects of vasoactive intestinal polypeptide. AB - Cyclic AMP accumulation in rat superior cervical ganglia during synaptic activity occurs by a noncholinergic, nonadrenergic process. Both preganglionic nerve stimulation and 4-aminopyridine increase ganglion cyclic AMP levels in the presence of atropine or phentolamine. Of the polypeptides tested as putative transmitters, vasoactive intestinal polypeptide (10(-6) M) causes ganglion cyclic AMP accumulation comparable to that produced by preganglionic nerve stimulation. PMID- 6288863 TI - Isolation of cell surface membranes from cultured C6 glioblastoma cells. AB - Plasma membranes were isolated from C6 glioblastoma cells by two methods. In the first method cells were treated with concanavalin A and lysed in hypotonic medium. After partial separation of plasma membranes from other cell material, the lectin was displaced with alpha-methyl-D-mannoside. In the second method untreated cells or cells iodinated in a lactoperoxidase-catalyzed reaction were homogenized in isotonic medium. Membrane fractions obtained by either homogenization procedure were further purified by rate zonal and equilibrium centrifugations into linear density gradients. Disruption of the glioblastoma cell membrane gives rise to heterogeneous assemblies of membrane fragments. Two populations of plasma membranes were isolated from untreated and from iodinated cells: a "lighter" membrane fraction characterized by relatively lower sedimentation velocity and buoyant density, and a "heavier" membrane fraction of relatively faster sedimentation velocity and higher buoyant density. Both fractions showed electrophoretic patterns similar to those of 125I-labeled cell surface proteins. Their specific (Na+ + K+)-ATPase activity was seven- to eightfold the homogenate activity (recovery, 13.1%). Both fractions were, however, still contaminated by smooth endoplasmic reticulum, as judged from the activity of NADPH-dependent cytochrome c reductase (recovery, 2.4%). It is suggested that plasma membrane fragments present in the two fractions might differ in the organization of their structures, e.g., membrane vesicle intactness and membrane orientation. PMID- 6288864 TI - An analysis of GABA receptor changes in the discrete regions of mouse brain after acute and chronic treatments with morphine. AB - The effects of morphine on the affinity and distribution of GABA receptors in the mouse regions (striatum, medulla, diencephalon, cortex, and cerebellum) were investigated in relation to: (a) acute administration, (b) chronic administration (tolerance), (c) precipitated withdrawal by naloxone, an opiate antagonist, and (d) abrupt withdrawal for 8 and 24 h. The alterations in the affinity as reflected by the dissociation constant (KD) and the number of receptors (Bmax) in the synaptic membranes obtained from controls and various treatments were determined by radioligand binding assay using [3H]muscimol as a ligand. Significant changes were observed in striatum, medulla, and diencephalon, whereas other regions including whole brain exhibited marginal changes. In general the number of GABA receptors increased after tolerance development, which upon abrupt withdrawal returned to control levels except in the case of naloxone-induced precipitated withdrawal. The affinity changes in different regions were diverse in nature and were not evident in the whole brain membranes. These results indicate that: (as) the regional alterations in the affinity and distribution of GABA receptors may play a role in the induction, maintenance, and regression of morphine tolerance; (b) abrupt withdrawal and antagonist precipitated withdrawal affect the GABA system differently, (c) chronic morphine treatment appears to influence the GABA receptors in the cerebellum, a region generally known for its lack of opiate receptors. PMID- 6288865 TI - Purification and characterization of bovine brain 5'-nucleotidase. AB - The 5'-nucleotidase located in the cytoplasmic fraction of bovine brain cortex was purified to electrophoretic homogeneity. The molecular weight was 134,000 daltons in the presence of sodium deoxycholate, whereas the enzyme formed high molecular weight aggregates in the absence of detergent. The purified enzyme showed the same kinetic and electrophoretic behaviour as the enzyme present in the original cytoplasmic fraction, and the presence of surfactants did not change the Km and Vm values. The nucleotidase from this source was a phosphohydrolase of 5'-mononucleotides acting on the deoxyribonucleotides and ribonucleotides of purines and pyrimidines. 5'-IMP was the preferred substrate; the optimum pH was 7.5. The study of the influence of the temperature on the initial reaction rates allowed calculation of the delta Ea and delta H degrees values. The variation of Vm and Km with a change in pH suggests the existence of a sulfhydryl group and an imidazole group in the enzyme-substrate complex. PMID- 6288866 TI - Determination of dopamine and its acidic metabolites in brain tissue by HPLC with electrochemical detection in a single run after minimal sample pretreatment. AB - A method, based on reverse-phase liquid-liquid chromatography, has been developed for the determination, in a single run, of dopamine (DA) and its acidic metabolites 3,4-dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA), combined with electrochemical detection (ECD). If applied to brain tissue, sample pretreatment can be reduced to centrifugation, filtration and adjustment of pH and perchlorate concentration prior to introduction into the liquid chromatograph. The relation between the perchlorate (counterion) concentration of the mobile phase and the retention (k') of the amines is linear, as is the relation between the H+ concentration of the mobile phase and the retention of the acidic metabolites. This flexible phase system, combined with a simple and therefore reproducible sample pretreatment, warrants a high throughput of samples. The procedure offers good possibilities for routine analysis of catecholamines and their acidic metabolites in the picogram range. Some typical examples of the behaviour of this phase system and the electrochemical detector are presented and discussed. PMID- 6288867 TI - P-31 nuclear magnetic resonance analysis of brain: the perchloric acid extract spectrum. PMID- 6288868 TI - Receptors for vasoactive intestinal polypeptide on isolated synaptosomes from rat cerebral cortex. Heterogeneity of binding and desensitization of receptors. PMID- 6288869 TI - Intrinsic protein phosphorylation in synaptosomal plasma membrane fragments: a comparison of cerebral cortex tissue from several species, including human biopsy specimens. AB - Intrinsic protein phosphorylation was studied in synaptosomal membrane fragments made from cerebral cortex tissue taken from the following species: human (biopsy specimens), ox, rat, rabbit, guinea pig and mouse. Membrane fragments from all species exhibited a qualitatively similar range of protein acceptors phosphorylated by cyclic AMP-dependent protein kinase activity; contrary to a previous report, no evidence for cyclic GMP-dependent protein kinase activity was found in the human material. With the exception of membrane fragments prepared from ox brain, all the preparations exhibited the same range of Ca2+-dependent protein kinase activity. Ox brain obtained from a slaughterhouse yielded membranes containing no Ca2+-dependent protein kinase activity, but this may have been due to unavoidable postmortem losses. PMID- 6288871 TI - Effects of Na+, Ca2+, and acetylcholine on phosphoinositide- and ATP-phosphate turnover in 32P-labeled rabbit iris smooth muscle. AB - Parallel studies were carried out in the rabbit iris on (a) the effects of Na+ and/or Ca2+ on the acetylcholine-stimulated 32P labeling of phosphatidic acid (PA) and phosphatidylinositol (PI) and the breakdown of polyphosphoinositides (poly PI), and (b) the effects of these cations on the specific radioactivity of [gamma-32P]ATP. Incorporation of 32P1 into ATP and phosphoinositides is time dependent, and it is remarkably dependent upon Na+ concentration in the incubation medium. The Na+ effect is reversible. Calcium ion, in the absence of Na+, had no effect on the specific radioactivity of ATP in 32P-labeled iris muscle; however, it moderately stimulated the 32P labeling of PA and PI and the breakdown of poly PI. In contrast, the addition of Na+, in the presence or absence of Ca2+, significantly reduced the specific radioactivity of ATP and 32P labeling of phospholipids in the 32P-labeled iris muscle. Acetylcholine had no measurable effect on the specific radioactivity of ATP. Furthermore, the neurotransmitter stimulated the 32P labeling of PA and PI and the breakdown of poly PI in the 32P-labeled muscle only in the presence of both Na+ and Ca2+. These data provide additional support for the concept that in the rabbit iris receptor-activated Ca2+ fluxes mediate or precede the effects of alpha-adrenergic and cholinergic muscarinic agents on phosphoinositide breakdown into 1,2 diacylglycerol and inositol phosphates and that restoration of the polar head groups to the 1,2-diacylglycerol (i.e., the recovery stage) is probably associated with Na+ outflux, via the Na+ -pump mechanism. PMID- 6288870 TI - Glycine antagonists structurally related to muscimol, THIP, or isoguvacine. AB - Microelectrophoretic methods were used to study the effects on cat spinal neurones of a number of compounds structurally related to the gamma-aminobutyric acid (GABA) agonists muscimol, THIP, and isoguvacine. While N-methylmuscimol was an agonist at bicuculline methochloride-sensitive GABA receptors, somewhat weaker than GABA and THIP, neither N,N-dimethylmuscimol nor N-methyl-THIP interfered significantly with GABA receptors in vivo or binding sites in vitro. Both N,N dimethylmuscimol and N-methyl-THIP, however, reversibly antagonized the depressant action of glycine. The seven-membered ring analogues of THIP, namely THIA (5,6,7,8-tetrahydro-4H-isoxazolo[5,4-c]azepin-3-ol), THAZ (5,6,7,8 tetrahydro-4H-isoxazolo[4,5-d]azepin-3-ol) and iso-THAZ (5,6,7,8-tetrahydro-4H isoxazolo[3,4-d]azepin-3-ol), also blocked neuronal inhibition by glycine, iso THAZ being the most potent compound. The conformationally mobile isomer of THAZ and iso-THAZ, 3-PYOL (5-(3-pyrrolidinyl)-3-isoxazolol), was a much less selective glycine antagonist, being also an antagonist of GABA, 3,4-TAZA (2,5,6,7 tetrahydro-1H-azepine-4-carboxylic acid) and 4,5-TAZA (2,3,6,7-tetrahydro-1H azepine-4-carboxylic acid), which are amino acid analogues of THIA and THAZ, respectively, and ring homologues of isoguvacine, were also shown to be glycine antagonists. The mechanism of action of the present class of zwitterionic glycine antagonists is unknown. The compounds are much less potent than strychnine. PMID- 6288873 TI - Phosphoproteins of the adrenal chromaffin granule membrane. AB - A fraction of chromaffin granule membranes contained a number of substrates for endogenous protein kinase activity as well as endogenous phosphatase activity. The major 32P-labelled polypeptide of molecular weight 43,000 appeared to be the alpha-subunit of pyruvate dehydrogenase of residual mitochondria. Several polypeptides showed cyclic AMP stimulation of phosphorylation of which the major polypeptide of molecular weight 59,000 shows half-maximal phosphorylation with 0.49 microM cyclic AMP. The phosphorylation of several other polypeptides is inhibited at high cyclic AMP concentrations. From studies with immunoprecipitation and two-dimensional electrophoresis it was found that alpha- and beta-tubulin and actin were absent from the granule membranes. However 32P labelling of a proportion of the copies of dopamine-beta-hydroxylase was demonstrated. The majority of the substrates for endogenous protein kinase activity are probably on the cytoplasmic side of the granule membrane. PMID- 6288872 TI - High specific binding of [3H]GABA and [3H]muscimol to membranes from dendrodendritic synaptosomes of the rat olfactory bulb. AB - Olfactory bulbs contain dendrodendritic synapses, which occur between granule cells and mitral cells, and gamma-aminobutyric acid (GABA) is thought to act as an inhibitory neurotransmitter at these synapses. Synaptosomes derived from the dendrodendritic synapses of the olfactory bulb were shown previously to contain considerable L-glutamate decarboxylase activity. The subcellular distribution and binding parameters of [3H]GABA and [3H]muscimol binding sites have now been determined in the rat olfactory bulb. Of all fractions examined, crude synaptic membranes (CSM) prepared from the dendrodendritic synaptosomes were shown to have the highest specific binding activity and accounted for nearly all of the total binding activity for both ligands. The specific binding activities for [3H]GABA and for [3H]muscimol were greatly increased after treating the CSM with 0.05% Triton X-100. Binding was shown to be Na+-independent, reversible, pharmacologically specific, and saturable. High- and low-affinity sites were detected for both ligands, and both classes of sites had appreciably lower KD values for muscimol (KD1 = 3.1 nM, KD2 = 25.1 nM) than for GABA (KD1 = 8.6 nM; KD2 = 63.7 nM). The amounts of the high-affinity binding sites for muscimol and GABA were similar (Bmax = 1.7 and 1.5 pmol/mg protein, respectively). The results of the present experiments indicate that the GABA and muscimol binding sites represent the GABA postsynaptic receptor, presumably on mitral cell dendrites, and provide further support for the hypothesis that GABA functions as a neurotransmitter at the dendrodendritic synapses in the olfactory bulb. PMID- 6288874 TI - Acetylcholine release from isolated synaptic vesicles related to ionic permeability changes: continuous detection with a chemiluminescent method. AB - The effect of ionic permeability changes on acetylcholine (ACh) release from isolated cholinergic synaptic vesicles of Torpedo was studied using a chemiluminescent method for continuous ACh detection. Vesicles rendered freely permeable to potassium by valinomycin lost most of their ACh content in K+ media, if the accompanying anion was permeant; it thus appeared that ACh leakage occurred as the result of internal osmotic changes. Upon addition of ionophores that catalyse monovalent cation/H+ exchange (gramicidin D or a mixture of valinomycin plus protonophore FCCP), a rapid but transient ACh release was observed. Surprisingly, nigericin which also catalyses K+/H+ exchange, had no effect on ACh release. The divalent cation ionophore A23187 promoted ACh release only when calcium (and not magnesium) was introduced into the external medium in a millimolar concentration range. As the simultaneous addition of the protonophore FCCP and A23187 decreased this calcium-dependent ACh leakage, a releasing effect of A23187 through Ca2+/H+ exchange is suspected. The present results emphasise the role of internal protons for ACh retention inside synaptic vesicles. PMID- 6288876 TI - Ethylenediamine and GABA potentiation of [3H]diazepam binding to benzodiazepine receptors in rat cerebral cortex. AB - Specific binding of [3H]diazepam at a free concentration of 2 nM was found to be maximally potentiated by 117% in Tris-HCl buffer and 160% in Tris-citrate buffer by ethylenediamine (EDA), but only at relatively high concentrations of EDA (ED50 = 5 X 10(-5) M), although this potentiation was susceptible to a low dose (6 microM) of bicuculline. Dose-response curves show that EDA differs from GABA with respect to both potency and efficacy. In additivity experiments no evidence was found that EDA could act as a partial agonist at GABA receptors, and it was concluded that EDA and GABA apparently do not potentiate [3H]diazepam binding by acting on the same receptor. Scatchard analysis lends support to this hypothesis, indicating that the potentiation of [3H]diazepam binding by 3.16 X 10(-3) M EDA is due to an increase in receptor number (from 930 to 1170 fmol/mg protein) and not receptor affinity (remaining constant about 20 nM). Subsequent studies showed the potentiation to be reversible. It is concluded that EDA can act on the GABA benzodiazepine receptor ionophore complex but that this is probably not a direct action on the GABA receptor. It is suggested that EDA can be used to differentiate GABA receptors linked to benzodiazepine receptors from those not so linked. PMID- 6288875 TI - Distribution and localization of estrogen-sensitive dopamine receptors in the rat brain. AB - Administration of estrogen to adult male rats increases the density of striatal dopamine receptors. The densities of the dopamine receptors in the nucleus accumbens and cortex are not altered, while the density of those in the hippocampus is decreased. In the pituitary the density, on a whole pituitary basis, is not changed. The increased density of striatal dopamine receptors normally observed after estrogen treatment is prevented by prior injection into the striatum of kainic acid, which destroys the intrinsic neurons in the striatum. In addition, the benzodiazepine receptors in the striatum, cortex, hippocampus, and cerebellum are not altered by estrogen treatment, showing the specificity of the estrogen treatment and suggesting that the effects of estrogen are not mediated through benzodiazepine receptors. PMID- 6288877 TI - Noncorrelation of choline kinase or ATP citrate lyase with cholinergic activity in rat brain. AB - The activity of choline acetyltransferase was used as an index of cholinergic structures in regions of rat brain. The activities of ATP citrate lyase and choline kinase correlated poorly with cholinergic activity in whole tissue fractions, contrasting with the good correlation between acetylcholinesterase and choline acetyltransferase. Choline acetyltransferase was preferentially localised in synaptosomes prepared from regions of high (striatum) or intermediate (cortex, medulla oblongata/pons) cholinergic activity. In general, this was not true for either choline kinase or ATP citrate lyase. PMID- 6288878 TI - Differential expression of type A and type B monoamine oxidase of mouse astrocytes in primary cultures. PMID- 6288879 TI - Hair follicle discrimination dysfunction in multiple sclerosis patients. AB - A method was developed of assessing somatosensory deficits quantitatively using hair follicle displacement as a stimulus within a psychophysical signal detection task paradigm. Multiple sclerosis patients with and without somatosensory disturbances could be differentiated and compared with normal subjects. This method may distinguish patients with somatosensory dysfunction, and dorsal funiculus neuropathology may underlie this distinction. PMID- 6288880 TI - Abnormalities in the vagus nerve in canine acrylamide neuropathy. AB - Dogs exposed to acrylamide develop a sensorimotor peripheral neuropathy and megaoesophagus. The presence of neuropathy was confirmed electrophysiologically and histologically. Hindlimb motor conduction velocity was reduced and there was a loss of large diameter myelinated fibres in the dorsal common digital nerve and the tibial nerve. The conduction velocity of vagal motor fibres innervating the thoracic oesophagus was not decreased; there was a reduction in the conduction velocity of the mixed nerve action potential of the vagus. Degenerating nerve fibres were observed in the vagus in the midthoracic region. The damage to vagal nerve fibres may be an important factor in the causation of megaoesophagus. PMID- 6288882 TI - Severe peripheral neuropathy due to lithium intoxication. PMID- 6288881 TI - Anticonvulsant peripheral neuropathy: a clinical and electrophysiological study of patients on single drug treatment with phenytoin, carbamazepine or barbiturates. AB - Previous studies of phenytoin neuropathy in selected groups of chronic epileptic patients on polytherapy have indicated a widely varying incidence of clinical or electrophysiological abnormalities. In 51 previously untreated epileptic patients followed prospectively on phenytoin or carbamazepine monotherapy, assisted by blood level monitoring, for 1-5 years we found no clinical evidence of neuropathy. Eighteen per cent of the phenytoin group and none of the carbamazepine group had mild electrophysiological changes (abnormalities of sensory action potentials or sensory conduction). In the former group the occurrence of the electrophysiological abnormalities was possibly related to previous exposure to high phenytoin or low folate levels or both. In 10 chronic epileptic patients we demonstrated reversible slowing of sensory nerve conduction during phenytoin intoxication. In six selected epileptic patients on chronic barbiturate monotherapy we found clinical evidence of neuropathy in two and electrophysiological abnormalities in five, including reversible slowing of sensory conduction during intoxication in one. This suggests that barbiturate drugs may, like phenytoin, also contribute to anticonvulsant neuropathy. Careful monitoring of single drug therapy with avoidance of acute toxicity may reduce the risk of chronic anticonvulsant neuropathy. PMID- 6288884 TI - Two representations of the hand in area 4 of a primate. I. Motor output organization. PMID- 6288883 TI - Acute autonomic and sensory neuropathy associated with elevated Epstein-Barr virus antibody titre. PMID- 6288886 TI - Effect of cooling area 18 on striate cortex cells in the squirrel monkey. PMID- 6288885 TI - Comparison of response properties of cerebellar- and thalamic-projecting interpolaris neurons. PMID- 6288887 TI - Neocortical and caudate projections of intralaminar thalamic neurons and their synaptic excitation from midbrain reticular core. PMID- 6288888 TI - Reversibility of Ia EPSP investigated with intracellularly iontophoresed QX-222. AB - 1. Cat lumbosacral motoneurons were impaled by two individually advanced microelectrodes: one to record membrane potential (EM), the second to pass depolarizing currents. 2. During the passage of depolarizing current ramps the repetitive action-potential firing and the later high conductance (GM) state obscured and distorted Ia excitatory postsynaptic potentials (EPSPs) evoked by electrical stimulation of hindlimb muscle afferents. 3. Intracellular iontophoresis of QX-222 (a trimethyl analogue of lignocaine) or methylxylocholine, prevented action-potential generation and reduced the GM increase during current depolarization so that positive levels of EM could be reached. 4. Following QX-222 treatment it was possible to demonstrate a reversal of the Ia EPSP including its first part, at EM values between -13 and +32 mV. Reversal was seen in 13 of the 22 motoneurons tested. 5. Reversal was easiest to obtain in motoneurons of the deep peroneal group. More positive levels of EM were needed to show a reversal in neurons of the gastrocnemius-so-leus group. (The 10 90% rise times of the EPSPs were rather similar for both groups.) 6. In a few motoneurons the initial part of the Ia EPSP reversed at a more negative EM than a later part. This was best seen after subtraction of the extracellular field potentials from the records. PMID- 6288889 TI - Analysis of synaptic depression contributing to habituation of gill-withdrawal reflex in Aplysia californica. AB - 1. Repeated stimulation of the siphon skin results in short-term habituation of the reflex contractions of the gill (38). The habituation, in turn, is correlated with a depression of the excitatory postsynaptic potentials (EPSPs) in motor neurons from mechanoreceptor sensory neurons (SN) (7, 16). The present study was undertaken to examine the parametric features of the synaptic depression and gain insights into the mechanisms underlying the reduced transmitter release. 2. Single sensory neuron action potentials were repeatedly elicited with depolarizing current pulses while the amplitude of the resultant EPSPs in the motor neuron was monitored. Synaptic depression varies as a complex function of interstimulus interval (ISI). At an ISI of 1 s, depression is rapid and reaches a plateau at 36% of control. In contrast, the depression at an ISI of 100 s is less pronounced, showing a gradual decay to 65% of control with the 10th EPSP. Surprisingly, there are no significant differences in time course or magnitude of depression across a broad range of intermediate ISIs (3, 10, and 30 s), although depression at these ISIs is intermediate between the 1 and 100 s ISIs. 3. There is also a complex relationship between spike interval and the depression of the second of two EPSPs. Thus, depression of the second of two EPSPs or depression of a train of EPSPs is not a monotonic function of spike interval. Indeed, the data suggest that there may be a slight underlying facilitatory process with short spike intervals. 4. The results also indicate that the recovery of synaptic depression following a train of 10 stimuli is not constant. Shorter spike intervals produce more rapid recovery. 5. These data are inconsistent with a classical depletion model (33) for synaptic depression and indicate that either a single complex function of time and ISI or multiple functions underlie synaptic depression and its recovery at the sensory neuron synapse. PMID- 6288890 TI - Exogenous choline augments transmission at an identified cholinergic synapse in terrestrial mollusk Limax maximus. AB - 1. A variety of pharmacological tests indicate that the neuromuscular junction between the salivary burster neuron (SB) and the salivary duct muscle (SD) in the terrestrial mollusk Limax maximus is cholinergic. These include the effects of curare, atropine, and hexamethonium. 2. Exogenously applied choline can act both presynaptically and postsynaptically at the synapse between the SB and SD. a) When the SB-SD synapse is bathed in 30 microM choline, there is no measurable direct postsynaptic effect at the SB-SD synapse. After several hours of choline incubation, however, an increase is observed in the size of SB-elicited junction potentials recorded from the SD. b) When the synapse is bathed in 300 microM choline, a short-term decrease is seen in the amplitudes of junction potentials (JPs) recorded from the SD. This effect is interpreted as competition between choline and acetylcholine (ACh) for postsynaptic receptor sites on the SD, since the response of the denervated SD to ACh is diminished in the presence of 300 microM choline. c) After several hours incubation in 300 microM choline, the amplitudes of JPs elicited by the SB on the SD increase. 3. The long-term effect of exogenous choline is blocked by the choline-uptake inhibitor, hemicholinium 3. 4. We conclude that exogenous choline is taken up by the presynaptic terminals of the SB and converted to ACh. Transmitter output rises with growing transmitter stores, producing an increase in the size of SB-elicited JPs recorded from the SD. PMID- 6288891 TI - Dietary choline augments blood choline and cholinergic transmission in terrestrial mollusk Limax maximus. AB - 1. A radioenzymatic assay was used to determine the concentration of choline in the hemolymph of the terrestrial mollusk Limax maximus maintained on choline enriched and choline-deficient diets. The levels of free choline in the slug's hemolymph varied with the availability of choline in the diet. The concentration of choline in the plasma of choline-enriched animals averaged 5.5 microM while the mean circulating choline level of choline-deprived slugs was 3.3 microM. The difference in choline levels between the two groups was statistically significant. 2. Neurophysiological measurements of transmission at the isolated cholinergic synapses between the salivary burster neuron (SB) and the salivary duct muscle (SD) showed that this synapse was sensitive to fluctuations in the external choline concentration within the physiological range. Increases in exogenous choline from 0.5 to 2.0 microM, from 2.5 to 5.0 microM, and from 5.0 to 15.0 microM were followed by increases in the amplitudes of SB-elicited junction potentials recorded from the SD by a focal extracellular electrode. 3. Thus, the SB-SD synapse is sensitive to the fluctuations in exogenous choline that can occur with a change in diet. PMID- 6288892 TI - Neurite outgrowth and selection of new electrical connections by adult Helisoma neurons. PMID- 6288893 TI - Defective cyclic adenosine 3':5'-monophosphate phosphodiesterase in the Drosophila memory mutant dunce. AB - A detailed characterization of the cyclic nucleotide phosphodiesterase (PDEs) from normal Drosophila melanogaster was made, including purification of the two major enzymes to near homogeneity. A third more labile phosphodiesterase also was identified in crude homogenates. The total activity per fly of one of these three enzymes, PDE-II, is strongly influenced by the dunce locus. Two independently derived dunce mutants produce variations of PDE-II with modified intrinsic properties: a marked decrease of thermal stability in dunce and a 10-fold increase in the Michaelis kinetic constant in dunce. These defects, which persisted in purified preparations of PDE-II, were mapped genetically to dunce. The results support the identification of dunce as the structural locus for PDE II. The tight connection between the dunce gene and the PDE-II enzyme indicates that defective cyclic adenosine 3':5'-monophosphate metabolism is the primary lesion which leads to failure of dunce flies to learn in the olfactory associative conditioning paradigm of Quinn et al. (Quinn, W. G., W. A. Harris, and S. Benzer (1974) Proct. Natl. Acad. Sci. U. S. A. 71: 708-712). PMID- 6288894 TI - Single cholinergic receptor channel currents in cultured human muscle. AB - Single cholinergic channel currents were recorded in adult human muscle tissue culture. The agonists suberyldicholine and carbamylcholine produce channels with the same conductance as channels produced by acetylcholine but with different closing kinetics. The antagonist tubocurarine, alone or mixed with suberyldicholine, activates channels which close very rapidly. For agonist activated channels, the distribution of open state lifetimes shows deviations from the usual single exponential form. An excess of short duration openings indicates the presence of an additional faster kinetic process. The lifetime distribution data can be interpreted in terms of varying proportions of slow and fast components which are present in a ratio determined by curve-fitting the appropriate two-exponential function to observed open time distributions. This ratio shows great variability in muscle from older cultures, but the fast and slow time constants are relatively constant. The observation of double exponential open time distributions indicates that the mechanism of channel closing is more complicated than earlier evidence indicated. PMID- 6288895 TI - Binding of developing mouse cerebellar cells to fibronectin: a possible mechanism for the formation of the external granular layer. AB - The role of the matrix glycoprotein fibronectin in the formation of the external granular layer of the developing mouse cerebellum was investigated by in vitro studies of the binding of cerebellar cells to a fibronectin-coated culture substratum and by in vivo immunocytochemical localization of antiplasma fibronectin antiserum in cerebellar tissue. The adhesion of cells dissociated from embryonic and early postnatal mouse cerebellum is developmental stage specific when the cells are plated on tissue culture substrata derivatized with human plasma fibronectin. Cells dissociated from mouse cerebellum at embryonic day 13 form cellular aggregates on insoluble plasma fibronectin. In contrast, cells dissociated from embryonic day 16 through postnatal day 7 cerebellum form a monolayer. Time-lapse video recordings reveal extensive cell movement of late embryonic and early postnatal cerebellar cells on insoluble plasma fibronectin. Late embryonic and early postnatal cerebellar cells bind to fibronectin but do not degrade the fibronectin substratum. Immunocytochemical studies of the binding of antiplasma fibronectin antisera to cryostat sections of intact embryonic and early postnatal cerebellar tissue reveal a brightly stained region of endogenous fibronectin along the route of granule cell migration from the lateral caudal part of the neuroepithelium lining the fourth ventricle up onto the external surface of the cerebellar anlage. When the formation of the external granular layer is completed, the intense region of fibronectin is no longer visible. PMID- 6288896 TI - Autoradiographic localization of adenosine receptors in rat brain using [3H]cyclohexyladenosine. AB - Adenosine (A1) receptor binding sites have been localized in rat brain by an in vitro light microscopic autoradiographic method. The binding of [3H]N6 cyclohexyladenosine to slide-mounted rat brain tissue sections has the characteristics of A1 receptors. It is saturable with high affinity and has appropriate pharmacology and stereospecificity. The highest densities of adenosine receptors occur in the molecular layer of the cerebellum, the molecular and polymorphic layers of the hippocampus and dentate gyrus, the medial geniculate body, certain thalamic nuclei, and the lateral septum. High densities also are observed in certain layers of the cerebral cortex, the piriform cortex, the caudate-putamen, the nucleus accumbens, and the granule cell layer of the cerebellum. Most white matter areas, as well as certain gray matter areas, such as the hypothalamus, have negligible receptor concentrations. These localizations suggest possible central nervous system sites of action of adenosine. PMID- 6288897 TI - Septohippocampal cholinergic neurons are regulated trans-synaptically by endorphin and corticotropin neuropeptides. AB - The content of acetylcholine (ACh) in nerve terminals or the dorsal hippocampus was examined after intraventricular, intraseptal, or intrahippocampal administration of a variety of endorphin/corticotropin neuropeptides. beta Lipotropin, alpha-endorphin, gamma-endorphin, alpha-melanotropin, beta melanotropin, adrenocorticotropin-1-39 (ACTH1-39), and ACTH4-10 (1, 3, 10, or 30 micrograms each) did not affect levels of ACh in the hippocampus 30 min after injection into the lateral ventricle. beta-Endorphin, administered intraventricularly (1, 3, 10, or 30 micrograms) or intraseptally (1 microgram), increased levels of ACh, while ACTH1-24, injected similarly, decreased levels of the neurotransmitter. ACh concentrations remained unchanged after direct application of beta-endorphin or ACTH1-24 (1, 3, 10, of 30 micrograms each) into Ammon's horn. Acute unilateral transection of the fimbria/superior fornix resulted in a time-related decrease in hippocampal ACh concentrations. Levels of ACh did not change 1 hr after transection; however, concentrations of hippocampal ACh decreased significantly 1 d or 1 week after deafferentation. ACh levels in the contralateral hippocampus remained unaffected at all times tested. Fimbrial transection blocked fully both endorphin- and corticotropin-induced changes in hippocampal ACh after the neuropeptides were injected into the lateral ventricle or the septal region. Naloxone, which, after subcutaneous (1 mg/kg) or intraventricular (100 micrograms) injection alone, failed to change levels of hippocampal ACh, antagonized the effects of intraventricular or intraseptal beta endorphin or ACTH1-24 or hippocampal ACh levels. The results suggest a site of endorphin/corticotropin receptor interaction at the level of cholinergic cell bodies in the septal region for regulating the activity of septohippocampal cholinergic neurons. PMID- 6288898 TI - Age-related changes in the nervous system. PMID- 6288899 TI - Nursing management of the patient receiving high-dose BCNU with autologous bone marrow harvest. PMID- 6288900 TI - Infarct sizing with Tc-99m pyrophosphate. PMID- 6288901 TI - [Experimental inner ear pathology with herpes simplex virus infection II. Intralabyrinthine inoculation of herpes simplex virus in guinea pigs]. PMID- 6288902 TI - Immunizations: current controversies. PMID- 6288903 TI - Fatal intraoperative pulmonary embolization of Wilms tumor. PMID- 6288904 TI - Coxsackie B viruses and autoimmune diabetes. PMID- 6288905 TI - Metabolic requirements for the damage of Trypanosoma cruzi epimastigotes by human polymorphonuclear leukocytes. AB - Normal human polymorphonuclear leukocytes (PMN) are cytotoxic to T. cruzi epimastigotes sensitized with specific antiserum (T. cruzi + Ab). The damage follows an early phagocytic event, suggesting the intracellular destruction of the parasites. We have studied the characteristics of the killing using metabolic inhibitors of the effector cells. Oxygen consumption by PMN with unsensitized parasites was similar to the uptake by resting cells, but increased two- to fourfold when T. cruzi + Ab was used. This increase in O2 consumption, associated with phagocytosis of T. cruzi + Ab was not sensitive to 2 mM cyanide nor 100 microM azide. Addition of T. cruzi + Ab to human PMN also stimulated H2O2 production. When PMN were incubated with phenylbutazone, cyanide or azide, an inhibition of cytotoxicity against sensitized T. cruzi was observed. Under the same experimental conditions phagocytosis was unaffected. These results indicate that active oxygen reduction products and myeloperoxidase are involved in the destruction of sensitized, T. cruzi epimastigotes by normal PMN. PMID- 6288906 TI - Solubilization of the membrane-bound enzymes of the brush-border plasma membrane of Hymenolepis diminuta (Cestoda) using nonionic detergents. AB - The ability of nonionic detergents to solubilize the membrane-bound enzymes of the brush-border plasma membrane of Hymenolepis diminuta was investigated. Of the detergents tested (Triton X-100, Tween 80, Brij 35, Lubrol PX and WX, W-1, and beta-octyl-D-glucoside), only Triton was an effective solubilizing agent. Optimal solubilization was achieved by incubating an isolated fraction of the brush border membrane in the presence of 1% Triton X-100 for 60 min at 37 C, followed by centrifugation at 100,000 g for 60 min at 25 C. This treatment resulted in solubilization of 94% of the alkaline phosphohydrolase, 91% of the phosphodiesterase and ribonuclease, and 88% of the 5'-nucleotidase activities. The pH optima for enzymes solubilized in nonionic and ionic detergents (Triton and sodium dodecyl sulfate, respectively) did not differ. Isoelectric focusing of the Triton-solubilized material demonstrated the presence of at least 14 polypeptides, a majority of which had isoelectric points below pH 7. PMID- 6288907 TI - Analog inhibitors for the pyrophosphate-dependent phosphofructokinase of Entamoeba histolytica and their effect on culture growth. PMID- 6288908 TI - Neuron specific enolase: a marker for differential diagnosis of neuroblastoma and Wilms' tumor. AB - Thirteen human neuroblastoma and six Wilms' tumor biopsies have been analyzed for neuron specific enolase (NSE). THe relative activity of NSE in the neuroblastomas (including ganglioneuroblastoma and ganglioneuroma) ranged from 28% to 62.5% of total enolase activity. The corresponding figures for the Wilms' tumors were 1% to 4.5%. It appears that NSE can serve as a biochemical marker for neuroblastoma and be useful in the differential diagnosis of neuroblastoma and Wilms' tumor. PMID- 6288909 TI - WDHA syndrome caused by VIP-producing ganglioneuroblastoma. AB - A 3 yr 11 mo-old girl showing classical symptoms of WDHA syndrome was transferred to our department of surgery. In preoperative examination, serum vasoactive intestinal peptide (VIP) was markedly elevated and pancreatic tumor was suspected. However, no tumor was found in the resected pancreas. Unfortunately, an unexpected adrenal tumor (ganglioneuroblastoma) was found in autopsy. The tumor was judged VIPO positive, immunohistochemically. This case was thought to be WDHA syndrome caused by VIPO-producing ganglioneuroblastoma (VIPoma). PMID- 6288910 TI - The effect of denervation of the cyclic nucleotides levels of the vas deferens. AB - We measured the cyclic nucleotides in levels of the rat vas deferens surgically denervated in order to investigate the relationship between denervation supersensitivity and cyclic nucleotides. Two days after denervation, cyclic AMP slightly decreased to 90.4% of control and cyclic GMP also decreased but not significantly. So it is assumed that almost all of the cyclic nucleotides are not neurogenic but myogenic. When the rat vas deferens was incubated with l norepinephrine 10(-4) for 3 min, the cyclic GMP level in the vas deferens denervated for seven d was significantly higher than that in the intact specimen, and the cyclic AMP level in the intact one was almost the same as in the denervated preparation. Denervation produced leftward shift of the dose response curve in cyclic GMP level to l-norepinephrine. After incubation with acetylcholine (10(-4)M), however, cyclic GMP and cyclic AMP levels in the denervated vas deferens were almost the same as in the intact one. Therefore, it is suggested that cyclic nucleotides don't relate to the induction of nonspecific supersensitivity by denervation in the rat vas deferens. PMID- 6288911 TI - Relationship between the Ba-induced contraction and cyclic AMP levels in the isolated longitudinal smooth muscle from guinea pig ileum. AB - Relationship between contraction and cyclic AMP levels induced by BaCl2 was examined in the longitudinal smooth muscle isolated from guinea pig ileum. BaCl2 3 X 10(-3)M caused a fast initial contraction, often followed by a gradual decrease of the contractile state. There was an increase in the tissue cyclic AMP 7 min or 14 min after the application of Ba. A phosphodiesterase activator imidazole reinforced the later phase of contraction by Ba and inhibited the increase in cyclic AMP. These results indicate that there is still a positive correlation between relaxation and increase in cyclic AMP and that an inhibitory action mediated by cyclic AMP is veiled behind the Ba contraction. Furthermore, these results may be interpreted by assuming that strong Ba contraction operates a feedback mechanism and that the feedback mechanism is associated with cyclic AMP increase. Indomethacin, an inhibitor of prostaglandins synthesis, little influenced the Ba-induced increase in cyclic AMP and rather inhibited the Ba contraction. Propranolol, a beta-adrenergic blocking agent, failed to exert influence on the Ba contraction. Based on these facts, it is suggested that the increase in cyclic AMP is not mediated by prostaglandins or catecholamines. PMID- 6288912 TI - Effect of proteinase inhibitors having anti-inflammatory activity on gelatinase, elastase and cathepsin G isolated from rat polymorphonuclear leukocytes. AB - Gelatinase, elastase and cathepsin G isolated from the granule extract of rat polymorphonuclear leukocytes (PMNs) had similar properties to the enzymes of human PMNs already reported. Effect of proteinase inhibitors on these neutral proteinases isolated from rat PMNs was studied. Epsilon-Amino-n-caproic acid n hexyl ester, a proteinase inhibitor having anti-inflammatory activity, inhibited cathepsin G, whereas elastase was activated by the inhibitor. On the other hand, leupeptin, L-1-tosylamide-2-phenylethyl chloromethyl ketone and N-alpha-p-tosyl-L lysine chloromethyl ketone, which had been reported as anti-inflammatory inhibitors, had no inhibitory effect on these neutral proteinases. These results suggest that proteinase inhibitors reported as anti-inflammatory agents exert their anti-inflammatory actions not by direct inhibition of the neutral proteinases released from PMNs, but by other effects such as suppression of the infiltration of PMNs into inflammatory locus. PMID- 6288913 TI - Antigen-induced elevation of cyclic GMP level in plasma and its abolition in experimentally immunosuppressive mice. AB - Intravenous injection of antigenic dose of sheep erythrocytes (SRBC) into mice caused a 150-200% increase in plasma cyclic GMP level within 5 min which continued for 60 min thereafter. Immunization by soluble antigens such as dextran sulfate and bovine serum albumin also elevated cyclic GMP level in plasma. The plasma cyclic GMP increased by antigen stimulation might be derived from immunocompetent cells. This assumption was supported by several lines of evidence as follows: (1) rat erythrocytes which are less antigenic for mice caused a relatively low response in terms of increase in cyclic GMP compared with SRBC, (2) this cyclic GMP response was abrogated in animals under immunosuppressive states where X-irradiated, azathioprine-treated or tumor-bearing mice were used. Mice pretreated with agents which block autonomic nervous system functions develop normal plasma cyclic GMP responses upon SRBC injection. Our observation in this report and many in vitro studies by other investigators suggested that plasma cyclic GMP elevated by antigen stimulation may be mainly derived from lymphoid cells as a consequence of the triggering of immune response. PMID- 6288915 TI - Conformation of some disubstituted 9-acridanones. PMID- 6288914 TI - Solvent drag effect in drug intestinal absorption. I. Studies on drug and D2O absorption clearances. AB - It was shown that the intestinal absorption clearance of D2O (CLD2O) could be a more appropriate index to study the solvent drag effect than water volume flow which was the difference between water influx and outflux in the intestinal lumen. Then, the correlation between the intestinal absorption clearances of drugs (CLdrug) and CLD2O were studied using the in situ recirculating method in the rat small intestine. The drugs used were low molecular drugs, that is, benzoic acid, salicylic acid, p-hydroxybenzoic acid and antipyrine, and comparably high molecular drugs, that is, cephalexin (CEX), cefroxadine (CXD) and cephalothin (CET). CLdrug and CLD2O were obtained in hypertonic, isotonic and hypotonic perfused solution adjusted with sodium chloride. Consequently, the correlations for all drugs except CET were significant and high solvent drag effects were observed. CLdrug of benzoic acid, salicylic acid and antipyrine were approximately equal to CLD2O, suggesting that the intestinal mucosa could not distinguish these lower molecular drugs from water. For the high molecular drugs such as cephalosporins, however, some extent of reflection from the membrane was certainly found in CEX and CXD, and the extent in CET was assumed much larger than CEX and CXD, resulting that the contribution of solvent drag in CET could not be found. Consequently, it was suggested that the solvent drag had some important role in the intestinal absorption of cephalosporins. PMID- 6288916 TI - Quantification of the effect of excipients on bioavailability by means of response surfaces II: Amoxicillin in fat-silica matrix. AB - This report studies the bioavailability of amoxicillin in different fat-silica matrixes. A urinary excretion study was carried out on four formulations containing fat and silica excipients. The formulations were administered to 24 healthy volunteers according to a Latin-square design. The following percent proportions of fat-silica were used: 15:3.75, 15:7.50, 30:3.75, and 30:7.50. The urinary excretion curves were characterized using the quantity of unchanged drug excreted between 0-2 and 0-12 hr postadministration, respectively as parameters. The ANOVA results showed that both excipients had an additive effect on the quantity of drug excreted between 0 and 2 hr, whereas the effect on the quantity of drug excreted between 0 and 12 hr was also one of interaction between both excipients. Quantification of the ANOVA results in terms of excipient content was conducted by means of the adequate linear functions. At the same time, a dissolution study was carried out using the quantity of drug dissolved in 30 and 180 min as parameters. The behavior was similar to that encountered for the in vivo parameters. PMID- 6288917 TI - Buprenorphine: differential interaction with opiate receptor subtypes in vivo. AB - The mixed agonist-antagonist buprenorphine previously was shown to display a bell shaped dose-response curve with peak agonistic (antinociceptive) opiate effects at approximately 0.5 mg/kg s.c., 60 min after the dose and a gradual decline of the effects in the dosage range between 0.5 to 10 mg/kg (electrically induced vocalization test). In vitro, buprenorphine as well as the potent pure agonist etorphine possessed very high affinity for all of the opiate receptor subtypes that are labeled by the universal tracer [3H]diprenorphine. However, in vivo, [3H]buprenorphine and [3H]etorphine appeared to label preferentially a subset of the opiate receptor sites; the labeled subset may be identical to or include the mu receptor sites, since these sites were also labeled with high affinity by [3H]naloxone, a mu preferring antagonist ligand. Buprenorphine saturated this opiate receptor subpopulation in its agonistic dosage range (less than or equal to 0.5 mg/kg). Moreover, buprenorphine saturated the remaining receptor subpopulation, labeled in vivo by [3H]diprenorphine, over its antagonistic dosage range (0.5--10 mg/kg). Cooperative receptor binding behavior of buprenorphine was not detectable. These results are consistent with the hypothesis that noncompetitive autoinhibition occurs among the opiate receptor subtypes. PMID- 6288919 TI - Reduced beta adrenoceptor interactions of norepinephrine enhance contraction in the femoral artery from spontaneously hypertensive rats. PMID- 6288918 TI - Activation and blockade of cardiac muscarinic receptors by endogenous acetylcholine and cholinesterase inhibitors. AB - Cholinesterase inhibitors are known to potentiate the effects of acetylcholine (ACh) and vagal stimulation on the myocardium. The studies presented here demonstrate that cholinesterase inhibitors (ChEI) also have activity in isolated atria in the absence of extrinsic cholinergic stimulation and that, depending on the ChEI, either indirect stimulation or direct blockade of cardiac muscarinic receptors can occur. Muscarinic agonists inhibit cyclic AMP formation in atria and the ChEIs physostigmine, neostigmine and echothiophate likewise produce a marked attenuation of isoproterenol-stimulated cyclic AMP accumulation The effect of physostigmine appears to result from muscarinic receptor activation by endogenous ACh as it is blocked by atropine. In contrast, the ChEI ambenonium does not stimulate but instead blocks muscarinic receptors coupled to cyclic AMP accumulation. Radioligand binding studies provide direct evidence that both ambenonium and demecarium are relatively potent muscarinic receptor antagonists, whereas physostigmine and other ChEI have little direct receptor activity. Physostigmine and ambenonium also have different effects on heart rate in vivo, the former potentiating and the latter apparently blocking vagal tone. The inhibition of cyclic AMP formation produced by physostigmine can be used as a measure of the concentration of endogenous ACh available at muscarinic receptor sites. Physostigmine blocks cyclic AMP formation in atria incubated in the absence of calcium or in the presence of tetrodotoxin, suggesting that endogenous ACh is spontaneously released in the absence of neuronal activity or depolarization-secretion coupling. PMID- 6288920 TI - Indications for vascular alpha- and beta-2 adrenoceptors in synapses of the muscarinic pathway in the pithed normotensive rat. AB - In the pithed normotensive rat the adrenoceptors involved in the hypertensive and tachycardic effects of the indirectly acting sympathomimetic agent tyramine and of electrical stimulation of the spinal cord (TH5-L4 or C7-Th1) were analyzed. The tools used for the identification of the adrenoceptors were the selective alpha-1 adrenoceptor blocking drug prazosin, the selective alpha-2 adrenoceptor antagonist rauwolscine, the beta-1 blocker atenolol and the selective beta-2 adrenoceptor blocking agent ICI 118,551. The participation of vascular alpha-2 adrenoceptors in the pressor response of tyramine was shown. The increase in blood pressure induced by electrical stimulation of the spinal cord (TH5-L4) was used to demonstrate that alpha-2 adrenoceptors were activated via ganglionic muscarinic receptors. The tachycardia evoked by electrical stimulation of the spinal cord (C7-Th1) was not influenced by beta-2 adrenoceptor blockade. It was enhanced, however, by the alpha-2 adrenoceptor antagonist rauwolscine. It is hypothesized that activation of ganglionic nicotinic receptors leads to stimulation of the nearest varicosities interfering with alpha-1 adrenoceptors. However, activation of ganglionic muscarinic receptors may lead to an additional release of neurotransmitter in the more distant varicosities endowed with alpha-2 or beta-2 adrenoceptors. PMID- 6288921 TI - Desensitization to the inotropic effect of isoproterenol in cultured ventricular cells. AB - To determine whether monoalyers of cultured chick embryo ventricular cells would show tachyphylaxis to chronotropic and inotropic effects of a beta adrenergic agonist, spontaneously contracting monolayers of primary cell cultures were studied using a phase-contrast microscope-video motion detector system that permitted quantitation of the chronotropic and inotropic state. The monoalyers were chronotropically unresponsive to isoproterenol between 10(-9) to 10(-6) M and chronotropically unresponsive to a 6-fold increase in perfusate calcium concentration. However, the cells were very inotropically responsive to calcium and to isoproterenol. Expressing the isoproterenol inotropic response as a percentage of response to 3.6 mM Ca, the response to 10(-6) M isoproterenol was 79 +/- 4% of the Ca response and the EC50 for isoproterenol was 3 x 10(-9) M. The monolayers rapidly developed dose-dependent desensitization to the inotropic effect of isoproterenol; after a 30-min exposure to 1 x 10(-6) M isoproterenol, the inotropic response was 40 +/- 5% of the initial response; desensitization was long-lasting and could be prevented by propranolol. Response to calcium remained unchanged after exposure to 10(-6) M isoproterenol. Thus, the cultured cell preparation shows rapid, sustained, beta receptor specific desensitization to the inotropic effects of a catecholamine. PMID- 6288922 TI - Presynaptic opiate receptor-mediated inhibition of endogenous norepinephrine and dopamine-beta-hydroxylase release in the cat spleen, independent of the presynaptic alpha adrenoceptors. PMID- 6288923 TI - Is the red cell calcium pump electrogenic? AB - 1. In inside-out vesicles of high potassium permeability, prepared from human red cell membranes, volume changes accompanying the action of the Ca2+ pump were measured by recording the intensity of light scattered by a suspension of these vesicles. Replacing Cl- by the impermeant gluconate anion changed swelling into shrinking. 2. Assuming that in Cl- media two Cl- ions accompany one Ca+ ion moved by the pump and in gluconate media two K+ ions are exchanged for one Ca2+ ion resulted in a good agreement between relative Ca2+ transport rate obtained from the volume change and from direct measurement of 45Ca uptake in the two media. 3. The fact that it is possible to change co-transport of Ca with Cl- into counter transport of Ca2+ for K+ rules out that within the pump there is an obligatory coupling of Ca2+ movement with movement of another ion species (including the proton). The conclusion, therefore, is that the Ca2+ pump must be electrogenic. 4. The combination of measurement of volume change with direct measurement of 45Ca movement yielded 5-6 microliter/mg protein for the volume of the vesicles. PMID- 6288924 TI - Electrophysiological analysis of neuronal thermosensitivity in rat preoptic and hypothalamic tissue cultures. AB - 1. Ninety-six neurones in forty-six explant tissue cultures of rat medial basal hypothalamus (m.b.h.) and preoptic area (p.o.a.h.) were subjected to thermal stimulation. 2. Neuronal response to temperature was determined on the basis of an extrapolated Q10 calculated from the regression line relating mean spontaneous activity to bath temperature. 3. Thermal stimulation (28-41 degrees C) of p.o.a.h. cultures resulted in the identification of three distinct groups of neurones: (1) temperature-insensitive (0.5 less than or equal to Q10 less than or equal to 2), (2) warm-sensitive (Q10 greater than 2), and (3) cold-sensitive (Q10 less than 0.5). 4. No temperature-sensitive neurones were identified in m.b.h. cultures. 5. In the presence of a medium which effectively blocks synaptic transmission (12 mM-Mg2+ and 0.25 mM-Ca+) the sensitivity of both warm- and cold sensitive neurones was preserved. 6. These data indicate that thermosensitivity is a characteristic not only of the preoptic neuronal network in vitro but also is an intrinsic characteristic of individual neurones. PMID- 6288925 TI - Phasic bursting activity of rat paraventricular neurones in the absence of synaptic transmission. AB - 1. The purpose of this study was to determine whether the phasic bursting activity, characteristic of certain magnocellular neuropeptidergic neurones in rat hypothalamus, is dependent upon chemical synaptic input.2. Slices of hypothalamus were placed in an in vitro chamber with hippocampal slices. The synaptic response in the CA1 cell layer from Schaffer collateral stimulation was monitored before, during and after synaptic transmission was blocked by superfusion of medium containing high Mg(2+) (either 18.7 or 9.3 mM) and low Ca(2+) (0.05 mM). This well studied pathway was chosen as an assay of synaptic blockade because hypothalamic circuitry is relatively unknown.3. The electrical activity of twenty-two phasic bursting neurones in the lateral portion of the paraventricular nucleus (p.v.n.) was recorded. Nineteen of twenty-two phasic p.v.n. neurones were recorded only after synaptic transmission was blocked. The remaining three cells were firing phasically in standard medium when first encountered and continued to display phasic bursting activity for up to 1.25 hr after synaptic blockade. Active cells in nearby hypothalamic areas did not show phasic bursting patterns either before or after synaptic transmission was blocked.4. The phasic bursting activity of the p.v.n. neurones in this study and that of previously reported p.v.n. cells in vivo were similar in (a) firing rate within bursts (b) burst length and (c) silent period duration.5. It is concluded that phasic bursting in p.v.n. magnocellular neuropeptidergic cells is not dependent upon synaptically mediated excitation or recurrent inhibition as has been hypothesized earlier.6. Alternative hypotheses, based upon acute changes in [K(+)](o), endogenous membrane currents and electrotonic coupling are discussed as possible explanations of phasic bursting in these magnocellular neuropeptidergic cells. PMID- 6288926 TI - Induction and removal of inward-going rectification in sheep cardiac Purkinje fibres. AB - 1. In sheep cardiac Purkinje fibres superfused with K-free, Na-free medium, the membrane potential can be stable either at a low negative level (-50 mV) or at a high negative level (-100 mV). The mechanism underlying the existence of these two stable potential levels was investigated using the two-micro-electrode voltage-clamp technique.2. By applying a voltage clamp of a certain duration at an appropriate level the membrane potential could be shifted from one stable level to the other. The shift was observed in Cl-free medium, excluding a redistribution of Cl as a possible explanation.3. Currents during and following a voltage step and their change with amplitude and duration of the voltage step could not be explained on the basis of depletion or accumulation of K ions in the narrow extracellular clefts.4. Instantaneous currents determined from the high negative resting level showed a high conductance and a pronounced inward rectification, while measurements from the low negative resting level indicated a low conductance and absence of inward rectification. The steady-state current voltage relation was dependent on the holding potential and showed memory or hysteresis.5. Estimation of the conductance by superimposed short voltage-clamp pulses showed an increase in conductance during a hyperpolarizing clamp from the low negative level and a decrease in conductance during a depolarizing clamp from the high negative level. The time-dependent current during a hyperpolarizing clamp from the low negative level reversed direction at a potential level corresponding to E(K), assuming a cleft K concentration of about 1 mM. In the presence of 0.1 mM-Ba the time-dependent current was abolished.6. The results suggest that the shift between the two stable levels is due to a time-dependent conductance change in the K inward rectifier channel, i(K1). The existence of memory excludes activation or de-activation only depending on the voltage gradient. Interaction of extracellular K ions with a site in the membrane is proposed as the activating mechanism. PMID- 6288927 TI - The effects of vasoactive intestinal peptide on neuromuscular transmission in the frog. AB - 1. The effects of vasoactive intestinal peptide (VIP) on cholinergic transmission were studied at the neuromuscular junction of the frog.2. Bath application of VIP (10(-8)-10(-7) M) produced a dose-dependent increase in quantal content of the end-plate potential (e.p.p.), while having no effect on quantal size or on resting membrane potential of the muscle fibres.3. The increases in e.p.p. amplitude observed at end-plates blocked with Mg(2+) or curare were comparable.4. VIP increased miniature end-plate potential frequency when the nerve terminal was depolarized with K(+). This effect was abolished by removing Ca(2+) from the bathing solution.5. Ionophoretic application of VIP to the end-plate also increased quantal content, while having no effect on quantal size or on the input resistance and resting membrane potential of the fibres.6. Some possible mechanisms for the action of VIP are discussed, and it is proposed that it may act on some step involved in depolarization-secretion coupling. PMID- 6288928 TI - Modulation on neuromuscular transmission by endogenous and exogenous prostaglandins in the guinea-pig mesenteric artery. AB - 1. At concentrations of 2.8 x 10(-8)-2.8 x 10(-6) M, prostaglandins (PGs; PGE(1), PGE(2) and PGF(2alpha)) had no effect on membrane potential and resistance of smooth muscles of the guinea-pig mesenteric artery. PGs (2.8 x 10(-8) M) suppressed the contraction evoked by perivascular nerve stimulation, but did not suppress the contraction evoked by direct muscle stimulation.2. PGs (2.8 x 10(-8) M) suppressed the e.j.p.s evoked by repetitive perivascular nerve stimulation but preserved the facilitation process of e.j.p.s evoked by any given stimulus frequency (0.1-1.0 Hz).3. The relationships between e.j.p. amplitudes and [Ca](o) plotted on log scales in the presence or absence of PGE(2) were not parallel. High concentrations of [Ca](o) prevented the inhibitory actions of PGs on the amplitude of e.j.p.s.4. PGE(2) did not suppress the activity of nerve fibres contributing to the generation of e.j.p.5. Indomethacin (10(-6) M) enhanced the amplitude of e.j.p.s and the frequency of miniature e.j.p.s with no change in the membrane potential and resistance of smooth muscles; these actions of indomethacin were suppressed by PGE(2) (2.8 x 10(-8) M).6. Phentolamine (10(-7) M) enlarged and yohimbine (10(-7) M) reduced the amplitude of the first e.j.p. evoked by a train stimulation, but the maximum amplitude of e.j.p., after the facilitation was completed, was in both cases much larger than that observed in the control. The enhancement of the transmission process was also suppressed by PGs (2.8 x 10(-8) M).7. The results indicate that in the guinea-pig mesenteric artery, PGs mainly suppress chemical transmitter release from nerve terminals due to interactions with Ca influx, but not due to interaction with presynaptic alpha adrenoceptors. Endogenous PG may act as a regulator substance in neuromuscular transmission. PMID- 6288929 TI - Calcium channel and calcium pump involved in oscillatory hyperpolarizing responses of L-strain mouse fibroblasts. AB - 1. In fibroblastic L cells, spontaneously repeated hyperpolarizing responses (oscillation of membrane potential) and hyperpolarizing responses evoked by electrical stimuli were suppressed by the external application of a K(+) channel blocker, nonyltriethylammonium (C(9)). This hydrophobic TEA-analogue also inhibited the hyperpolarization induced by intracellular Ca(2+) injection.2. Quinine or quinidine, known inhibitors of the Ca(2+)-activated K(+) channel of red cells, instantaneously inhibited these hyperpolarizations. Thus, these hyperpolarizations are likely to be caused by the operation of Ca(2+)-sensitive K(+) channels.3. Azide, which is known to inhibit the mitochondrial Ca(2+) uptake in fibroblasts, and caffeine, dantrolene Na and oxalate, which affect the microsomal Ca(2+) transport, did not exert any effects upon the electrical potential profiles.4. On the other hand, Ca(2+) channel blockers (nifedipine, D 600 and Co(2+)) suppressed the hyperpolarizing responses, but not the hyperpolarizations produced by intracellular Ca(2+) injection, suggesting that the calcium ions responsible for the hyperpolarizing responses are mainly derived from outside the cell through Ca(2+) channels.5. Flavones of plant origin, which are known to inhibit Ca(2+)-ATPase, prolonged the duration of the hyperpolarizing phase of the oscillation or produced a sustained hyperpolarization.6. It is concluded that the Ca(2+) channel and the Ca(2+) pump play essential roles in the generation of the hyperpolarizing response and of the membrane potential oscillation in L cells, and that these hyperpolarizations are brought about by a transient elevation of cytosolic Ca(2+) level which, in turn, activates Ca(2+) dependent K(+) channels. PMID- 6288930 TI - An ionophoretic study of the responses of rat caudal trigeminal nucleus neurones to non-noxious mechanical sensory stimuli. AB - 1. Extracellular recordings of the responses of single caudal trigeminal nucleus neurones to non-noxious and noxious facial stimuli and to ionophoretically applied L-glutamate, L-aspartate and acetylcholine were made in urethane anaesthetized rats. 2. Neurones excited by non-noxious mechanical stimuli were located primarily in the magnocellular part of nucleus caudalis, whereas neurones excited by both noxious and non-noxious stimuli were located either ventromedially to the magnocellular part of nucleus caudalis or superficially to the substantia gelatinosa. 3. Both L-aspartate and L-glutamate were found to excite all neurones tested in nucleus caudalis. In contrast, however, acetylcholine was found to excite only 31% of the neurones tested. 4. Responses of nucleus caudalis neurones to non-noxious sensory stimulation were not antagonized by the excitatory amino acid antagonist D-alpha-aminoadipate, but were antagonized by cis-2, 3-piperidine dicarboxylate and gamma-D glutamylglycine, two excitatory amino acid antagonists with a broader spectrum of action. 5. It is concluded that the chemical synaptic transmitter of non nociceptive mechanoreceptive primary afferent fibres to nucleus caudalis may be a ligand for an excitatory amino acid receptor other than a D-alpha-aminoadipate sensitive receptor. The synaptic receptor may thus be of the kainate or quisqualate type, and the transmitter possibly L-glutamate, L-aspartate or an as yet unidentified substance. PMID- 6288931 TI - Combined effects of calcium and dibutyryl cyclic AMP on germinal vesicle breakdown in the mouse oocyte. AB - Mouse oocytes were cultured in the presence of dibutyryl cyclic AMP (dbcAMP) and various agents that affect cytoplasmic calcium concentrations. Treatment that inhibited calcium uptake potentiated the inhibitory effect of dbcAMP and treatments which stimulated cellular calcium uptake overcame the effect of dbcAMP. Elevated extracellular calcium (greater than 10 mM) significantly decreased the inhibitory effect of concentrations of dbcAMP up to 150 microM when compared to control levels of calcium (1.7 mM). In addition, the calcium ionophore A23187 (greater than 1 microM) significantly overcame the effect of dbcAMP in media that contained 1.7 or 20 mM calcium. In the presence of 41 microM dbcAMP the calcium antagonist verapamil increased (in a dose-dependent fashion) the percentage of oocytes blocked at the germinal vesicle stage, from 21% with 10 microM-verapamil to 99% with 200 microM. A similar dose-dependent, reversible potentiation of the effect of dbcAMP was found with tetracaine, which also lowers cytoplasmic calcium concentrations. These results suggest that a minimum level of cytoplasm calcium is required for the initiation of germinal vesicle breakdown and that the action of dbcAMP is mediated by its effect upon this calcium. PMID- 6288933 TI - Autoradiographic analysis of changes in ovarian binding of FSH and HCG during induced follicular atresia in the hamster. AB - Immediately after hypophysectomy, 30 i.u. PMSG were injected s.c. and 3 days later an antiserum to PMSG was injected i.p. Groups of hamsters were killed at 0,24, 48 and 72 h after PMSG antiserum. The ovaries were prepared for topical autoradiography and the numbers of silver grains in different ovarian compartments were counted. The numbers of binding sites for 125I-labelled FSH in the granulosa cells of the antral follicles dropped sharply to 33, 14 and 5% of that at 0 h at 24, 48 and 72 h respectively. Binding of 125I-labelled hCG to granulosa cells declined more slowly, being 47, 27 and 24%, respectively. Binding of 125I-labelled hCG to thecal and interstitial cells was unaffected. Compared to other models of atresia, the changes in gonadotrophin binding observed in this model occur at an accelerated rate because of the acute deprivation of PMSG. PMID- 6288934 TI - Oestradiol administration raises luteal LH receptor levels in intact and hysterectomized pigs. AB - Occupied and unoccupied LH receptors in corpora lutea, and LH and progesterone concentrations in circulating plasma, were measured in non-pregnant gilts that had been treated with oestradiol-17 beta benzoate to prolong luteal function. Oestradiol benzoate (5 mg, administered on Day 12 after oestrus) delayed luteal regression and the decline in LH receptor levels at luteolysis and raised unoccupied receptor levels from 11.8 +/- 1.14 fmol/mg protein on Days 10--15 after oestrus to 31.8 +/- 3.26 fmol/mg protein on Days 15--21. There was no simultaneous rise in occupied receptor levels and occupancy decreased from 29.8 +/- 3.01 to 11.5 +/- 1.26%. Basal plasma LH concentrations were unchanged by oestradiol, but mean corpus luteum weight and plasma progesterone concentrations were slightly reduced. Oestradiol benzoate on Day 12 caused a similar increase in unoccupied receptor levels in gilts hysterectomized on Days 6--9 after oestrus, from 17.0 +/- 5.83 to 34.5 +/- 6.00 fmol/mg protein, determined on Days 15--18. Plasma concentrations of LH and progesterone were unchanged by oestradiol. Unoccupied receptor levels in corpora lutea and plasma LH and progesterone were unaltered by hysterectomy in untreated gilts. Occupied receptor levels were not influenced by hysterectomy or oestradiol. It is concluded that oestradiol-17 beta raises luteal LH receptor levels by a mechanism independent of the uterus. PMID- 6288932 TI - Relationship between LH receptor concentrations in thecal and granulosa cells and in-vivo and in-vitro steroid secretion by ovine follicles during the preovulatory period. PMID- 6288935 TI - Testicular FSH receptor numbers and affinity in bulls of various ages. AB - Bulls (N = 42) ranging in age from 1 day to 5.5 years were used to determine whether a change in the concentration of FSH receptors in the bovine testis occurred as bulls matured. 125I-labelled human FSH was used as the ligand to evaluate binding to bovine testicular membranes. Membrane fractions were collected by centrifugation of testicular homogenates at 120 g and recentrifugation of the 120 g supernatant at 1250 g. Relative binding activity of membrane sedimented at 1250 g was determined after incubation of membranes with 125I-labelled FSH for 16--18h at 25 degrees C, followed by centrifugation (1250 g) to separate bound from free hormone. Specifically bound FSH when expressed as fmol/mg protein was negatively correlated with age (r = -0.73). The association constant (Ka) determined by Scatchard analysis was the same for bulls at all ages with a mean (+/- s.e.m.) Ka = 1.5 +/- 0.3 x 10(9) M-1. Concentration of FSH receptors on a per mg protein basis declined rapidly from birth to 2.5 years of age and remained low up to 5.5 years of age. On a whole testis basis the total number of receptors increased as the bulls matured. After 2.5 years of age total testicular binding did not change. PMID- 6288936 TI - Gestational trophoblastic disease. The effect of duration of disease and hCG titer on response to therapy. AB - The height of the pretreatment hCG titer and the time interval from termination of the antecedent pregnancy to institution of treatment were determined in 352 patients with gestational trophoblastic disease in order to judge their effect, both individually and together, on response to therapy. When all patients in need of treatment for gestational trophoblastic disease, both metastatic and nonmetastatic, were considered as one group, examination of time alone, of hCG titer alone and of time and titer together each permitted the identification of patients at high risk with equal reliability (p less than 0.0005 for each). When patients with only metastatic gestational trophoblastic disease were evaluated, time and titer taken separately and together each identified those patients at high risk, but not in an equal manner (time alone, p = 0.02; titer alone, p less than 0.05; time and titer together, p less than 0.0005). Time and hCG titer, alone or in combination, did not have a statistically significant effect on outcome when patients with metastatic choriocarcinoma were considered separately. Other factors, such as metastatic site and antecedent pregnancy, seem to be more important in determining prognosis than duration of disease and hCG titer in this group of patients. PMID- 6288937 TI - A quantitative assay of phagocytosis using liposomes with trapped spin labels. PMID- 6288938 TI - Reduction in cyclic 3',5'-adenosine monophosphate phosphodiesterase activity in exudate and cultured mouse peritoneal macrophages. AB - Cyclic nucleotides have been implicated in the normal function of macrophages, but the nature of the cyclic nucleotide catabolizing enzyme phosphodiesterase (PDE) and its role in regulation of macrophage function has not been previously reported. Cyclic adenosine monophosphate PDE activity was studied in mouse peritoneal macrophages and found to be similar to that reported in other tissues with regard to stability and divalent cation requirement. Exudate macrophages induced by injection of either endotoxin or proteose peptone broth were found to have reduced PDE activity and enhanced EAC phagocytic capacity and acid phosphatase (AP) activity. When resident peritoneal macrophages were cultured in vitro; AP activity increased over 24 hr and remained elevated for 96 hr. PDE activity declined steadily over 96 hr. This steady decline in PDE activity observed during culture was not altered by inclusion of endotoxin. These studies suggest that PDE activity is reduced in macrophages with maximally enhanced function but does not appear to be significantly altered during early phases of functional change in macrophages. PMID- 6288939 TI - Pretreatment with phorbol myristate acetate inhibits macrophage activity against intracellular protozoa. AB - To further document the role of toxic oxygen intermediates in mononuclear phagocyte antiprotozoal activity, microbicidal macrophages were depleted of the capacity to generate superoxide anion (O-2) and hydrogen peroxide (H2O2) by pretreatment with phorbol myristate acetate (PMA), a soluble agent which triggers the macrophage respiratory burst. Treating cells for 90 min with 200 ng/ml of PMA inhibited the extracellular release of both O-2 and H2O2 by 90% upon subsequent restimulation with either PMA or opsonized zymosan. This effect persisted for 48 h, and could not be reversed by the addition of lymphokine. Intracellular nitro blue tetrazolium reduction by PMA-treated cells was also inhibited by 66-95% upon rechallenge with either PMA or inert or viable particulate agents. In parallel, PMA pretreatment abolished or markedly impaired the ability of normal, lymphokine stimulated, and in vivo activated macrophages to kill three diverse protozoa- Toxoplasma gondii, Leishmania donovani, and Trypanosoma cruzi. These studies illustrate an additional technique for investigating the antiprotozoal effects of macrophage-derived O-2 and H2O2 and reemphasize the importance of an intact respiratory burst mechanism in killing of intracellular parasites. PMID- 6288940 TI - Pseudosecondary antibody response to LPS induced by toxins for macrophages. AB - A single dose of bacterial lipopolysaccharide, administered 21 days after mice had been treated with the macrophage toxins carrageenan, or microparticulate crystalline silica, resulted in a secondarytype antibody response. The phenomenon of pseudosecondary responsiveness was investigated to determine the mechanism for its generation. The results showed that generating a pseudosecondary response was T cell dependent; but except for this, it followed the same kinetic and genetic patterns as a true, two dose of antigens, secondary response. It was concluded that pseudosecondary responsiveness resulted from priming the mice by the consequential effects of the macrophage toxins; thereafter, secondary responsiveness to LPS was generated in a normal manner. PMID- 6288941 TI - Depressed blood neutrophil response to lithium stimulation in newborn rats. AB - The blood neutrophil response to lithium chloride challenge was determined in one day-old and young adult rats to test the hypothesis that granulocytopoiesis is less efficient in neonates than in adults. Adult animals responded rapidly to lithium chloride treatment, showing a 134% increase in blood neutrophils on the second day of exposure and ultimately increasing by 260% by the tenth day. Newborn rats on the other hand, showed a 25% decrease in blood neutrophils after one day of treatment with lithium, then remained at that or at pretreatment levels throughout the subsequent 10 days. These observations suggest that hematopoietic tissue in the newborn is less responsive than is that of adults. PMID- 6288942 TI - The effect of auranofin and sodium aurothiomalate on peripheral blood monocytes. AB - The effect of oral and parenteral gold preparations on the functional capacity of human peripheral blood monocytes was studied in various in vitro systems. Both agents were capable of inhibiting monocyte chemotaxis, expression of Fc and C3 receptor sites and to a lesser extent the reduction of nitroblue tetrazolium dye. No difference was observed in the effect of oral or parenteral gold on normal and rheumatoid monocytes. The inhibition of monocyte function by gold salts was associated with an increase in intracellular cyclic AMP levels. Our data suggests that the beneficial effect of gold salts in rheumatoid arthritis may result from its action on the functional activities of the monocyte-macrophage system. PMID- 6288943 TI - Serum Angiotensin-converting enzyme (SACE) in temporal arteritis. PMID- 6288944 TI - Malignant mixed tumour of the liver and its response to adriamycin. PMID- 6288945 TI - Potential affinity labels for the opiate receptor based on fentanyl and related compounds. AB - Derivatives of fentanyl, 3-methylfentanyl, sufentanil, and lofentanil, possessing chemo- or photoaffinity functionalities, were synthesized as potential affinity reagents for the opiate receptor. Opiate receptor binding constants (IC50) were determined in competition experiments with [3H]naloxone and [3H]naltrexone. Affinity-labeling experiments were generally unsuccessful, although some irreversible attachment was achieved with alpha-diazoamide 17 and aryl azide 23. PMID- 6288946 TI - (+/-)-2-Depentylperhydrohistrionicotoxin: a new probe for a regulatory site on the nicotinic acetylcholine receptor-channel. AB - (+/-)-2-Depentylperhydrohistrionicotoxin (4), several of its analogues, and N- and O-substituted derivatives were prepared and tested for their effects on the neuromuscular transmission of the frog sartorius muscle. Compound 4, its N-methyl derivative 5, the O-acetyl derivative 9, and the quaternary methiodides 19 and 20 blocked the indirectly elicited twitch. The oxidation of 4 and 5 to ketones 12 and 14 and their reduction to the epimeric alcohols 17 and 18 afforded materials with substantially reduced activity. N-Acetylation of 4 to 11 changed the course of the activity to a transient potentiation of muscle twitch. Both 4 and 5 were not very toxic to mice after subcutaneous administration. (+/-)-7-n-Butyl-1 azaspiro[5,5]undecan-8-one (12) epimerized readily at room temperature to afford the epimer 13, and preparation of the hydrochloride of its N-methylated derivative 14 was accompanied by a retro-Michael reaction, affording the 2-n butyl-3-[4-(methylamino)butyl]cyclohexene-2-one (22). The strongly hydrogen bonded alcohol 4 was analyzed as the hydrobromide by a single-crystal X-ray analysis, confirming its structure. PMID- 6288947 TI - Angiotensin converting enzyme inhibitors: modifications of a tripeptide analogue. AB - Modified nonhydrolyzable tripeptide analogues of (S)-1-[5-(benzoylamino)-1,4 dioxo-6-phenylhexyl]-L-proline (1), designed to impart oral angiotensin converting enzyme (ACE) inhibitory activity, were made and evaluated in vivo and in vitro. The N-methyl and C5-methyl analogues of 1 were inactive. Insertion of heteroatoms (O, S, NH) into the C--C chain of 1 gave a series of compounds with high in vitro activity in the guinea pig serum ACE assay. The O-analogue was the most potent with an IC50 = 4.4 x 10(-9) M compared to 1 with an IC50 = 3.2 x 10( 9) M. The structure-activity relationships in this series of compounds lead one to speculate that the heteroatom provides an additional binding site to the surface of the enzyme; however, these compounds were inactive when tested for antihypertensive activity in the renal hypertensive rat at 30 mg/kg by the oral route (captopril is active at 1.0 mg/kg po). PMID- 6288948 TI - Familial Poland anomaly. AB - The Poland anomaly is usually a non-genetic malformation syndrome. This paper reports two second cousins who both had a typical left sided Poland anomaly, and this constitutes the first recorded case of this condition affecting more than one member of a family. Despite this, for the purposes of genetic counselling, the Poland anomaly can be regarded as a sporadic condition with an extremely low recurrence risk. PMID- 6288949 TI - Micromelia, polysyndactyly, multiple malformations, and fragile bones in a stillborn child. PMID- 6288950 TI - Age-related changes in the prevalence of precipitating antibodies to BK virus in infants and children. AB - Nearly 1000 sera from children were tested by immunoelectro-osmophoresis against BK virus, and age-specific prevalence rates were estimated from birth until the age of 12 years. Declining rates during the first 12 months showed the waning of passive immunity, which at birth reflects the mother's immune status. The changes of prevalence suggested that the peak incidence of primary infections occurred at about 2 years, with an estimated peak annual rate of 24.6%. PMID- 6288951 TI - Differentiation of subtypes within Leptospira interrogans serovars Hardjo, Balcanica and Tarassovi, by bacterial restriction-endonuclease DNA analysis (BRENDA). AB - Various strains of Leptospira interrogans were compared by bacterial restriction endonuclease DNA analysis (BRENDA). Field strains of serovar hardjo isolated from domestic animals in New Zealand, Australia and Northern Ireland were indistinguishable from one another but differed strikingly from the hardjo reference strain Hardjoprajitno. Similarly, field isolates of balcanica and tarassovi differed from their serovar reference strains, probably owing to a difference in epidemiological niche. Subdivision of these serovars into distinct subtypes as defined by BRENDA is therefore useful and justified. In contrast, analysis of serovars pomona, ballum and copenhageni shows that field and reference strains were identical, or differed only by a single band. It is suggested that BRENDA will overcome many of the problems associated with serological methods of identifying serovars and allow more precise definition of epidemiological relationships between strains and their hosts. PMID- 6288952 TI - Binding of type-III group-B streptococci to buccal epithelial cells. AB - A binding assay was used to study the attachment of type-III group-B streptococci (GBS) to buccal epithelial cells. Results indicate that an adhesin, with the characteristics of a protein, is the molecule at the streptococcal cell surface responsible for attachment to the buccal cells. The bacterial adhesin probably recognises a sugar on the surface of the mucosal cell, because periodate oxidation of the buccal cells caused a significant reduction in subsequent adherence of GBS. A sonicate to type-III GBS blocked the binding of the organism to buccal cells. The effects of physical and chemical modifications of the sonicate on its ability to prevent bacterial attachment are described; these corroborate the evidence gained from heat and periodate treatments of the buccal cells and GBS. Results suggest a lectin type of attachment mechanism for type-III BGS which can be blocked by N-acetyl-D-glucosamine, rather than attachment by means of a lipoteichoic acid as described for group-A streptococci. PMID- 6288953 TI - Influence of sulpiride-induced hyperprolactinemia on baboon menstrual cycles: a longitudinal study. AB - The influence of sulpiride-induced hyperprolactinemia on the hypothalamic pituitary-ovarian function of the baboon (Papio cynocephalus) was investigated. Plasma levels of prolactin, LH, FSH, estrone, estradiol, 17-hydroxyprogesterone, progesterone and 20 alpha-dihydroprogesterone in control and consecutive treatment cycles (sulpiride i.m. injections 100 mg/day) were determined serially. The hormonal changes indicate that the ovary is the most sensitive site to the direct inhibitory action of sulpiride-induced hyperprolactinemia. PMID- 6288954 TI - Rotavirus antibodies in hanuman langurs (Presbytis entellus). AB - Serum samples from wild Hanuman langurs (Presbytis entellus) from Mysore State, India, were compared to samples from a laboratory colony from Davis, Calif., for antibodies to rotavirus, which is an important cause of gastroenteritis in mammals. Animals from the laboratory colony had a higher frequency and higher levels of antibody than wild animals. It is likely that wild populations of langurs have a much lower incidence of rotaviral infection than laboratory populations, which are exposed to both crowded conditions and rotaviruses from other species. PMID- 6288955 TI - Herpesviral pneumonia and septicemia in two infant Gelada baboons (Theropithecus gelada). PMID- 6288957 TI - Progesterone-induced down-regulation of electrogenic Na+, K+-ATPase during the first meiotic division in amphibian oocytes. AB - Progesterone initiates the resumption of the meiotic divisions in the amphibian oocyte. Depolarization of the Rama pipiens oocyte plasma membrane begins 6-10 hr after exposure to progesterone (1-2 hr before nuclear breakdown). The oocyte cytoplasm becomes essentially isopotential with the medium by the end of the first meiotic division (20-22 hr). Voltage-clamp studies indicate that the depolarization coincides with the disappearance of an electrogenic Na+, K+-pump, and other electrophysiological studies indicate a decrease in both K+ and Cl- conductances of the oocyte plasma membrane. Measurement of [3H]-ouabain binding to the plasma-vitelline membrane complex indicates that there are high-affinity (Kd = 4.2 x 10-8M), K+-sensitive ouabain-binding sites on the unstimulated (prophase-arrest) oocyte and that ouabain binding virtually disappears during membrane depolarization. [3H]-Leucine incorporation into the plasma-vitelline membrane complex increased ninefold during depolarization with no significant change in uptake or incorporation into cytoplasmic proteins or acid soluble pool(s). This together with previous findings suggest that progesterone acts at a translational level to produce a cytoplasmic factor(s) that down-regulates the membrane Na+, K+-ATPase and alters the ion permeability and transport properties of both nuclear and plasma membranes. PMID- 6288956 TI - Characteristics of antibodies to guinea pig (Na+ + K+)-adenosine triphosphatase and their use in cell-free synthesis studies. AB - Antibodies have been produced, in three rabbits, to Na/K-ATPase purified from guinea pig renal outer medulla. Each rabbit produced antibodies to both the alpha (catalytic) and the beta (glycoprotein) subunits of Na/K-ATPase. The titers of the anti-alpha and anti-beta antibodies varied with time and between rabbits. None of the antisera inhibited Na/K-ATPase activity under various preincubation conditions. A method is presented for separating small amounts of anti-alpha subunit from anti-beta subunit antibodies. There was no cross-reactivity of antibodies to one subunit with the other subunit. The alpha subunit of the Na/K ATPase was cleaved into a 41,000-dalton peptide (that contains the ATP phosphorylating site) and a 58,000-dalton hydrophobic peptide as described by Castro and Farley (Castro, J., Farley, R.A., 1979, J. Biol. Chem. 254: 2221 2228). Anti-alpha antibodies from all of the rabbits reacted with both proteolytic fragments. The anti-guinea pig Na/K-ATPase antisera (pooled) cross reacted with the alpha subunit of Na/K-ATPase from human, cow, dog, rabbit, rat, mouse, turtle, and toad; and with the beta subunit from human, rat, and mouse. The loci of cross-reactivity were investigated using partially purified canine kidney Na/K-ATPase cleaved with trypsin as described above. The anti-sera from rabbits 1 and 2 cross-reacted with the 41,000-dalton peptide from the dog but very little with the 58,000-dalton peptide. No cross-reactivity was observed with antiserum from rabbit 3 to either fragment. Guinea pig kidney RNA was translated in a rabbit reticulocyte lysate system followed by immunoprecipitation with the antisera. The molecular weight of the cell-free synthesized alpha chain was 96,000 daltons. Its identity was established with purified anti-alpha antibodies and by immunocompetition with purified Na/K-ATPase and Ca-ATPase. Translation of the beta subunit was not detected in this system. PMID- 6288958 TI - Anomalous electrophoretic mobility of restriction fragments containing the att region. PMID- 6288959 TI - Regulatory mutants of simian virus 40. Effect of mutations at a T antigen binding site on DNA replication and expression of viral genes. PMID- 6288960 TI - Chromosomal position effects determine transcriptional potential of integrated mammary tumor virus DNA. PMID- 6288961 TI - Pathway of vesicular stomatitis virus entry leading to infection. PMID- 6288962 TI - The 3'-non-coding regions of alphavirus RNAs contain repeating sequences. PMID- 6288964 TI - Calculation of the electric potential in the active site cleft due to alpha-helix dipoles. PMID- 6288963 TI - Binding of an analog of the simian virus 40 T antigen to wild-type and mutant viral replication origins. PMID- 6288965 TI - Crystallization of resolvase, a repressor that also catalyzes site-specific DNA recombination. PMID- 6288966 TI - A new class of mutants in DNA polymerase I that affects gene transposition. PMID- 6288967 TI - Chemical evolution 40. Clay-mediated oxidation of diaminomaleonitrile. PMID- 6288968 TI - Acceleration of HCN oligomerization by formaldehyde and related compounds: implications for prebiotic syntheses. PMID- 6288969 TI - Antibody to hepatitis A virus in healthy Nigerians. AB - Two hundred and fifty Nigerians (175 male and 75 female), whose ages ranged from 5 to 70 years, were surveyed for the presence of antibody to hepatitis A virus (anti-HAV). The prevalence was determined to be 82 percent. The rates were highest in blood donors (90 percent) and hospital workers (91.4 percent) and lowest in children under the age of 10 (25 percent). The prevalence rates were not related to socioeconomic groups, previous exposure, jaundice, or sex. The study confirms that HAV infection is endemic in Nigeria and that most infections are subclinical and occur early in life. Acute hepatitis in a Nigerian adult may therefore not be due to HAV. PMID- 6288970 TI - Toxicokinetics of 2,2',4,4',5,5'-hexabromobiphenyl in the rat. PMID- 6288971 TI - Investigation of hyperkeratotic activity of polybrominated biphenyls in Firemaster FF-1. AB - A polybrominated biphenyl (PBB) mixture was fractionated by normal-phase preparative high-performance liquid chromatography. The hexane fractions were concentrated and applied to rabbit ears. Only the most polar fraction produced hyperkeratosis on the rabbit ears. This active fraction was subfractionated by using the same procedure. Again, the extent of hyperkeratotic activity increased with increasing polarity. The PBBs of the largest concentration levels in the active fraction were purified by preparative gas chromatography and tested on rabbit ears. The major compounds did not demonstrate hyperkeratotic activity. PMID- 6288972 TI - Synchronously occurring malignant fibrous histiocytoma of the kidney with contralateral renal cell carcinoma. PMID- 6288973 TI - The occurrence of Wilms tumor in 2 patients with exstrophy of the bladder. PMID- 6288974 TI - Importance and timing of prophylactic antibiotics in urology with a special reference to growth and kill rates of E. coli in genitourinary organs. AB - We conducted an experimental study using rats to investigate the timing of prophylactic antibiotics in urological surgery and to evaluate the benefit from a single, prophylactic injection of an antibiotic. Another objective was to examine the growth and kill rates of Escherichia coli 06 in operated urinary tract organs. We were unable to demonstrate a short decisive period for the antibiotic treatment; it seems to vary with respect to the infecting organism and the infected organ. Administration of a single dose of antibiotic was sufficient to lower the rate of infection in the kidney and epididymis 24 hours after the operation and to eliminate bacteria completely from the prostate. The rate of growth and kill of E. coli 06 depended on the specific organs in which the infection took place. PMID- 6288976 TI - Cefotaxime failure in group A streptococcal meningitis. PMID- 6288975 TI - Phosphodiesterase activity of the lower urinary tract. AB - Cyclic AMP is believed to mediate the physiologic response of beta-receptor stimulation in the lower urinary tract. A possible novel therapy for specific dysfunctions might be via pharmacologic modification of cyclic AMP through regulation of phosphodiesterase, the enzyme responsible for metabolizing cyclic AMP. The present study characterizes the phosphodiesterase activity present in various sections of the lower urinary tract and determines the effect of calmodulin and the potency of a series of phosphodiesterase inhibitors. The results can be summarized as follows: the phosphodiesterase activity of all smooth muscle sections of the lower urinary tract were similar. They showed nonlinear kinetics with Vmax between 1.0 and 2.4 nmol./mg. protein/minute and apparent Km's around 100 microM. The external sphincter (skeletal muscle) displayed linear kinetics with a Vmax of 0.2 nmol./mg. protein/minute and a Km of 9 microM. Of the drugs tested, papaverine was the most potent inhibitor and theophylline was one of the weakest inhibitors. These studies also indicate that the major form of cyclic AMP phosphodiesterase is not sensitive to calmodulin. PMID- 6288977 TI - Implication of plasma free hemoglobin in massive clostridial hemolysis. AB - Prolonged tissue oxygenation and maintenance of intravascular oncotic pressure are severely impaired with the absence of an effective RBC mass. We recently encountered a patient with a nondetectable hematocrit value (0%) attributed to Clostridium perfringens hemolysis. The patient maintained normal BP, tissue oxygenation, and mentation and survived longer than four hours, despite ineffective transfusion therapy. Plasma free hemoglobin was responsible for the preservation of tissue oxygenation, intravascular oncotic pressure, and pH. Existing studies in animals support stroma-free hemoglobin as an effective blood product in humans. PMID- 6288978 TI - Chlamydia trachomatis and cervical neoplasia. AB - We tested 383 women with and 500 women without cervical neoplasia for antibodies against Chlamydia trachomatis or herpes simplex virus (HSV). Exposure to both agents was related to sexual activity, with the highest prevalence of antibodies found in women with more sex partners and who had first coitus at an earlier age. When subjects were matched for several risk factors (age, race, marital status, parity, number of sex partners, and history of venereal disease), a significant excess of antibodies against C trachomatis was found in cases as compared with control subjects (76.5% v 58.4%, respectively; n = 149). Because matched-pair analysis lost a substantial proportion of women with neoplasia (largely because they were older), linear logistic analysis was performed. This also showed an excess of antichlamydial antibody in cases, with an estimated odds ratio of approximately 2 for the association of antichlamydial antibody and the risk of being a case. Neither analysis found an excess of antibodies to HSV type 2 in cases. PMID- 6288979 TI - [Endorphins]. PMID- 6288980 TI - [Mechanism of diarrhea and WDHA syndrome]. PMID- 6288981 TI - [Gastrointestinal hormone secreting tumor]. PMID- 6288982 TI - [Gastrin and GRP]. PMID- 6288983 TI - HLA-D typing using Epstein-Barr virus-induced lymphoblastoid cell lines. AB - We examined whether Epstein-Barr virus-induced lymphoblastoid cell lines, established from homozygous typing cells (HTC-EBV-LCL), could be substituted for normal homozygous typing cells (HTC) in HLA-D typing. When using EBV-LCL as stimulators in one-way MLR, the stimulator: responder ratio was found to be most important. At a ratio of 1:10, the autologous MLR between EBV-LCL and PBL from the same donors exhibited low double normalized values (DNV) which could be distinguished from those of homologous combinations. The panel cells typed with HTC always exhibited low DNV in repeat one-way MLR in which EBV-LCL established from the same HTC were used. On the other hand, by employing our previously described method, we were able to establish the necessary HTC-EBV-LCL from unselected seropositive adult donors and utilize these cells for HLA-D typing. We suggest that our method makes it possible to circumvent the problem presented by the shortage of HTC, since our HTC-EBV-LCL can be substituted for normal HTC in HLA-D typing. PMID- 6288984 TI - [Effect of aging on the concentrations of arachidonic acid and eicosapentaenoic acid in human serum lipids]. PMID- 6288985 TI - [Effect of hemodialysis on pituitary-adrenal function]. PMID- 6288986 TI - [The role of renal adrenergic receptor on renin secretion after the stimulation with parasympathomimetic drug]. PMID- 6288987 TI - Recent advances in clinical application of blockers of renin-angiotensin system. PMID- 6288988 TI - 19-hydroxyandrostenedione, a new amplifier of the action of aldosterone, in low renin essential hypertension. PMID- 6288989 TI - Sheep lung carcinoma: an endemic analogue of a sporadic human neoplasm. PMID- 6288990 TI - Major histologic types of cancers of the gum and mouth, esophagus, larynx, and lung by sex and by income level. AB - Incidence rates by income level were computed for squamous cell carcinomas of the gum and mouth, larynx, and esophagus and for squamous cell carcinoma, small cell carcinoma, and adenocarcinoma of the lung in white males and females aged 35--64 years, with the use of data for the nine geographic areas of the Third National Cancer Survey and the 1970 U.S. census. Within sex, age, and geographic area groups, patterns of cancer incidence by income level were analyzed by use of a nonparametric statistical method. Higher rates for males than for females were found for every histologic type studied, and the ratio of male-to-female rates increased with age (especially for squamous cell carcinomas of the larynx and lung). A strong inverse relation to income level was found for all of the histologic types studied in males and for squamous cell carcinomas of the gum and mouth and esophagus and small cell carcinoma of the lung in females. These findings are discussed with reference to patterns of smoking and alcohol use by sex and social class. PMID- 6288991 TI - Cerebrospinal fluid markers in African Burkitt's lymphoma with central nervous system involvement. AB - Several cerebrospinal fluid markers were found to be elevated in Burkitt's lymphoma patients with central nervous system (CNS) involvement. Antibody levels to the virus capsid antigen of the Epstein-Barr virus and to the brain cell antigens myelin and cerebroside were elevated during active CNS disease. Immune complexes were present in levels above 100 micrograms/ml in most patients with CNS involvement but tended to be low or negative in patients without CNS disease. Oligoclonal IgG bands were present in 12 of 13 patients with CNS disease and in only 3 of 26 patients with no clinical evidence of disease. None of these markers were present in 6 other tumor patients without CNS disease. The presence of these markers in 12 of 13 patients in whom CNS disease was involved suggests that these markers may be useful in determining the status of the tumor with regard to involvement of the CNS. PMID- 6288992 TI - Transforming growth factors in the urine of normal, pregnant, and tumor-bearing humans. AB - Transforming growth factor (TGF) activities could be detected in the urine of normal, pregnant, and tumor-bearing humans. These acid- and heat-stable polypeptides competed for binding to epidermal growth factor (EGF) membrane receptors and promoted the anchorage-independent growth of nontransformed rodent cells. They differed from human EGF in their apparent molecular weights and soft agar growth-stimulating activity. The urine from pregnant females contained TGF activities with apparent molecular weight(s) (relative) (Mr) of 10,000 ad 17,000- 20,000. In the case of a lung cancer patient, an additional major activity of approximately 30,000--35,000 Mr was found. All urine specimens examined also contained a "common" 8,000-Mr soft-agar growth-stimulating activity, which competed for binding to EGF membrane receptors and which was chromatographically separable from EGF (urogastrone). Thus urine may provide a convenient and readily available source for the biochemical characterization of these TGF-like activities, some of which may be clinically useful biologic markers for certain types of cancer. PMID- 6288993 TI - Simian virus 40 (SV40)-specific isoelectric point-4.7--94,000-Mr membrane glycoprotein: major peptide homology exhibited with the nuclear and membrane associated 94,000-Mr SV40 T-antigen in hamsters. AB - Tryptic peptide maps of electrophoretically purified 94,000-molecular weight (relative) (Mr) nuclear and membrane-associated simian virus 40 (SV40) T antigens, TN and TM, respectively, were compared to those of the SV40-specific isoelectric point (pI)- 4.7--94,000-Mr plasma membrane component reactive with anti-T-sera from Syrian golden hamsters. Bidimensional thin-layer electrophoresis and chromatography of TN labeled with 125I revealed about 27 tryptic peptides. A similar number of peptides was identified for TM and the pI-4.7--94,000-Mr component. A peptide homology between TN and TM or TN and the pI-4.7-94,000-Mr protein exists and indicates that the previously described pI-4.7--94,000-Mr membrane component represents TM. Only 4 of 27 peptides were labeled when TM was subjected to lactoperoxidase-catalyzed radioiodination from the outer surface of the plasma membrane. One of these TM peptides was metabolically labeled with [14C]glucosamine. The data indicate that TM is partially exposed on the cell surface and represents a glycosylated form of TN. Closely associated with TM is a pI-4.5--55,000-Mr membrane component. This component does not exhibit significant peptide homology with the 94,000-Mr SV40 protein and, therefore, appears to be coded for by the host cell genome. PMID- 6288994 TI - Epizootic neoplasms in fishes from a lake polluted by copper mining wastes. AB - Examination of fishes from Torch Lake, Houghton County, Mich., revealed epizootic neoplasms of several types in two closely related species. Saugers, Stizostedion canadense, and walleyes, Stizostedion vitreum, were commonly affected with hepatocellular carcinomas, dermal ossifying fibromas, and perivisceral masses resembling mesotheliomas that were usually associated with the mesenteric capsule of the spleen. Saugers were 100% affected with liver neoplasms. Histopathologic and ultrastructural aspects of the neoplasms are described. Torch Lake has been used as a repository for copper mining waste to the extent that an estimated 20% of the original volume has been filled in by these materials. Several direct and indirect etiologic roles for the mine waste as causative agents of the fish tumors are discussed. PMID- 6288995 TI - Comparison of biologic behavior and histology of transplanted mammary tumors in GR mice. AB - Mammary tumors that arose spontaneously in inbred GR mice were transplanted into syngeneic castrated males. The hormone responsiveness of the transplants was studied in mice treated with estrone and progesterone and was compared with the hormone responsiveness in mice that received no hormone treatment. Microscopic examination of hormone-responsive and hormone-independent tumors revealed similar histologic patterns in both groups. It was evident that pale cells, which are classically associated with hormone-responsive tumors, may also be present in transplanted hormone-independent tumors in this strain. No correlation was found between the histologic pattern of these transplanted tumors and the biologic behavior, hormonal status, or presence of a specific murine mammary tumor virus (MuMTV) proviral fragment. Mammary tumors also appeared capable of undergoing differentiation into more than one morphologic type. Two cotransplanted tumors (derived from the same parental tumor) had markedly different histologic patterns; however, analysis of MuMTV proviral fragments indicated that the MuMTV infected cells were of the same parentage. PMID- 6288997 TI - [Medical topics: dietary fibers]. PMID- 6288996 TI - Secretagogue response in rat pancreatic acinar carcinoma. AB - The secretion of protein, like cell proliferation, is an integrated response that reflects structural organization of the cell periphery. Stimulation of protein secretion was thus utilized for comparison of integrated responses of the cell periphery in pancreatic acinar carcinoma of the rat and integrated responses in normal rat pancreas. Results of this comparison include: a) The stimulation of protein secretion in acinar carcinoma fragments by carbamylcholine chloride and cholecystokinin octapeptide, pancreatic secretagogues that interact with specific plasma membrane receptors, was only a fraction (one-fifth to one-half) of that observed in normal pancreatic minilobules. b) The Ca2+ ionophore A23187 and the cyclic nucleotide N6,O2'-dibutyryl cyclic AMP, secretagogues that act independently of specific membrane receptors, did not stimulate secretion in the acinar carcinoma. The observed quantitative and qualitative differences in protein secretion indicate fundamental differences in cell periphery organization between the normal and transformed acinar cells of pancreas. PMID- 6288998 TI - Multiple hormonal control of adenylate cyclase in distal segments of the rat kidney. AB - Using the single tubule adenylate cyclase microassay, we investigated in vitro in three different segments of the rat nephron whether the effects of various hormones are additive when these hormones are tested in combination. In the cortical portion of the thick ascending limb (CAL), no additivity of the effects of glucagon, calcitonin, and PTH was observed. In the medullary portion of the thick ascending limb (MAL), the effects of vasopressin and glucagon were only partly additive, and the effects of vasopressin and calcitonin were fully additive. In the cortical collecting tubule (CCT), the effects of calcitonin and vasopressin were nonadditive in the kidneys in which vasopressin alone induced a high cyclase stimulation, whereas they were fully additive when vasopressin induced a low cyclase stimulation. The data suggest that in each segment, the hormones tested stimulated the same cells: no additivity was observed when cyclase Vmax acted as the limiting factor of the response; partial or full additivity was observed when the number of hormone receptors acted as the limiting factor of the response. As a consequence, calcitonin, glucagon, and PTH should induce the same effects in CAL; vasopressin, glucagon, and calcitonin, the same effects in MAL; and vasopressin and calcitonin, the same effects in CCT. PMID- 6289000 TI - [Hormonal changes in septic endotoxic shock]. PMID- 6288999 TI - [Results of cardiac surgery and conclusions for medical after-care]. PMID- 6289001 TI - Plasma adenosine 3':5'--cyclic monophosphate response to glucagon in uremia. AB - The effect of a single, intravenously administered dose of glucagon on plasma cyclic adenoside monophosphate (cAMP) was studied in seven normal subjects, ten patients with chronic renal failure (CRF), and ten patients with terminal renal insufficiency (TRI) receiving long-term haemodialysis treatment (HD). Ten minutes following glucagon administration, uremic patients displayed a significantly (P less than 0.0001) greater increase in cAMP than control subjects. Glucose levels after glucagon administration did not differ significantly between the normal and uremic groups, and lipolysis was less pronounced in the uremic patients than in the controls (P less than 0.003). These results could not be attributed to differences in serum insulin response. The findings demonstrate differences in the hepatic adenylate cyclase and cAMP response between normal and uremic subjects. These alterations in cAMP responsiveness may play a role in the pathophysiology of the metabolic disturbances associated with uremia. PMID- 6289002 TI - Diurnal and ultradian variations of plasma concentrations of eleven adrenal steroid hormones in human males. AB - The diurnal variations of the plasma concentrations of eleven steroid hormones and of corticotropin (ACTH) were studied in ten young healthy males. The plasma steroids progesterone, pregnenolone, deoxycorticosterone, 17-OH-progesterone, 17 OH-pregnenolone, deoxycortisol, 18-OH-deoxycorticosterone, corticosterone, aldosterone, cortisol and 18-OH-corticosterone, as well as plasma ACTH, were measured at 30-min intervals in the morning and in the evening and at 2-h intervals during the rest of the day. Steroids were extracted from 1 ml plasma, fractionated by high-pressure liquid chromatography (HPLC) and finally quantified by radioimmunoassay (RIA). Plasma concentrations of ACTH were radioimmunoassayed after extraction from 2 ml plasma. More or less pronounced circadian and episodic variations were apparent for plasma levels of all steroids studied, as well as of ACTH. According to related profiles of diurnal variations of plasma concentrations, three different categories of steroids were tentatively crystallized. Category 1 includes 17-OH-pregnenolone, deoxycortisol, corticosterone, 18-OH-deoxycorticosterone, deoxycorticosterone, cortisol and 18 OH-corticosterone, exhibiting a rhythm partly synchronous with that of the pituitary secretory activity of ACTH. Category 2, including progesterone, pregnenolone and 17-OH-progesterone, exhibited a time course of plasma concentrations assuming a regulation predominantly dictated by the testicular secretory activity. Lastly, aldosterone exerted a variation of plasma concentrations which was obviously regulated by the renin-angiotensin system under the present conditions. PMID- 6289004 TI - [Clinicomorphological characteristics of suppurative wounds treated with iruksol]. PMID- 6289003 TI - Genetic disorders of leukocyte function: what they tell us about normal antimicrobial mechanisms of human phagocytic cells. AB - Analysis of three inherited defects of granulocyte function (Chediak-Higashi Syndrome, CHS; Chronic Granulomatous Disease, CGD; Myeloperoxidase Deficiency, MPO) has highlighted critical events for the antimicrobial function of these cells and placed others in perspective. Prompt phagosomal fusion may be more important for digestion of organisms rather than killing as indicated by the mild bactericidal defects in the CHS. The formation of O2- and H2O2 during the phagocytic respiratory burst is central for the broad antimicrobial activity of granulocytes. MPO, on the other hand, while perhaps normally participating in granulocyte antimicrobial action, appears to be essential only for the effective killing of eukaryotic organisms such as certain fungal strains. While the non oxidative killing mechanism of neutrophils have stimulated much recent interest and were the first to be defined no specific inherited defects have been discovered which are clinically important. Genetic disorders of macrophage effector function remain to be clearly defined as do those of eosinophils. The lessons learned from the study of the granulocyte defects discussed have provided both the technology and approach to the analysis of the antimicrobial and cytocidal mechanisms of these important phagocytic cells. PMID- 6289005 TI - [Lymphocyte beta-adrenoreceptor function in bronchial asthma treated conservatively and by surgery]. PMID- 6289006 TI - Peanut lectin-binding sites in large bowel carcinoma. AB - Peanut lectin is known to bind to B-D-Gal-(1 leads to 3)-D-GalNac which provides antigenic determination for the T (TAg) blood group antigen. We examined 33 rectosigmoid carcinomas and 15 corresponding controls for their ability to express peanut lectin-binding sites. In controls one could localize TAg to the supranuclear portion of the cell, however, in cancers one noticed a cytostructural relocalization of TAg with the following two major patterns: localization to the region of the glycocalyx and localization intracytoplasmically in the apical portion of the cell. These two patterns were associated with glandular differentiation. Less frequently noted or in association with the above was a mucin glob-like pattern and/or a fine diffuse intracytoplasmic pattern associated with solid, nonglandular areas. The more poorly differentiated cancers less frequently expressed peanut lectin-binding sites. Benign (nontransitional zone) epithelium in those patients whose tumor expressed TAg was negative for peanut lectin-binding sites in 66 per cent of the cases. Reduced tumoral glycosyltransferases may explain this increased synthesis of TAg in cancers as compared with controls, if one considers TAg to be an incomplete glycoprotein of the MN blood group system. PMID- 6289007 TI - Detection of type C virus particles in short-term cultured adult T cell leukemia cells. PMID- 6289008 TI - Identification of polybrominated biphenyls in the adipose tissues of the general population of the United States. AB - Hexabromobiphenyl has been identified by gas chromatography/mass spectrometry (GC/MS) in pooled extracts of adipose tissue samples collected from the general population of the conterminous United States. Mass spectra derived from tissue extracts subjected to gel permeation chromatography were compared with those obtained from an authentic PBB mixture containing 2,4,5,2',4',5' hexabromobiphenyl as the principal component. GC retention times and unique characteristics of the mass spectrum permitted confirmation of identity of the hexabromo isomer. Levels in one tissue sample were determined to be in the 1 to 2 ppb range. PMID- 6289009 TI - Refinement and validation of a model describing synthesis 'storage and release of acetylcholine at the neuromuscular junction. PMID- 6289010 TI - A simple method for the purification of rat brain Na+,K+-adenosine triphosphatase (ATPase). AB - Several methods of purification of Na+,K+-adenosine triphosphatase (ATPase) have been previously described for a wide variety of tissues. In general, highest activity preparations have necessitated large amounts of tissue and many purification steps. This article describes a technique that allows partial purification of Na+,K+-ATPase from as few as 15 rat brains and should be of interest to investigators of the pharmacology of this particular enzyme system. In this modified version of the Jorgensen procedure (Biochim Biophys Acta 356:36- 52, 1974) we purified the Na+,K+-ATPase from 15--90 rat brains, and obtained enzyme preparations with a mean specific activity of 552 +/- 37.6 mumol Pi/mg of protein/hr (95.5% ouabain sensitive). This "purified" enzyme had an activity ratio (Mg2+ + Na+ + K+)/(Mg2+ + Na+) of 47.4 +/- 12.3 SEM, compared to 3.29 +/- 0.17 SEM for the untreated microsomes. Ouabain inhibited the "purified" enzyme with an I50 of 6 X 10(-9) M. Ouabain binding (644 pmol/mg of protein) yielded a turnover number of 13,700 min-1. Sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis of the enzyme revealed predominantly the alpha and beta subunits with some minor contaminant bands. Previous methods of purification of rat brain Na+,K+-ATPase have employed sodium deoxycholate and high concentrations of NaI; the reported specific activity obtained was generally 150--350 mumol Pi/mg of protein/hr. We have employed higher SDS concentrations than in Jorgensen's technique for rabbit kidney but the procedure is simpler because sucrose gradients are not used. Final wash steps also include 10--20% glycerol in the media. These modifications have yielded Na+,K+-ATPase of significantly higher specific activity than previously reported for rat brain. PMID- 6289011 TI - Implication of the effect of extragenic territorial DNA sequences on a mechanism involving switch-on/off of neighbouring genes by transposable elements in eukaryotes. PMID- 6289012 TI - Informational aspects of Gibbs function output from nucleotide pools and of the adenylate kinase reaction. PMID- 6289013 TI - Surgical resection in the management of small cell carcinoma of the lung. AB - In an attempt to define the role of initial surgical resection in patients with undifferentiated small cell carcinoma of the lung, we reviewed the experience of the Veterans Administration Surgical Oncology Group (VASOG). One hundred forty eight patients with small cell carcinoma of the lung had undergone a potentially "curative" resection. This represented 4.7% of "curative" resections carried out in four major prospective adjuvant chemotherapy trials. In the early trials (101 patients), 16 patients (15.8%) died within the first 30 postoperative days. These patients have been excluded from the analysis of long-term survival, since in the more recent trials postoperative deaths were excluded prior to randomization. In the 132 patients remaining, the 5 year survival rate by the life-table method was 23.0%. The tumor of each was classified pathologically by the TNM system. Five year survival rates for each category were as follows: T1 N0 M0 59.9%, T1 N1 M0 31.3%, T2 N0 M0 27.9%, T2 N1 M0 9.0%, and any T3 or N2 3.6%. The effect of postoperative adjuvant chemotherapy was evaluated in each of the trials. No beneficial effect of the adjuvant therapy was noted with a one or two course regimen of either nitrogen mustard or cyclophosphamide, but possible benefit, although not significant, was noted in a prolonged intermittent chemotherapy trial of cyclophosphamide either alone or alternating with methotrexate. In the most recent trial of prolonged intermittent courses of 1-(2-chlorethyl)-3 cyclohexyl-l-nitrosourea (CCNU) and hydroxyurea, a 5 year survival rate of 80.8% was noted in those receiving adjuvant chemotherapy as compared to a 38.1% in the control group. We conclude that resection is definitely indicated in patients with T1 N0 M0 lesions and probably indicated in those with T1 N1 M0 or T2 N0 M0 lesions. Primary surgical resection is contraindicated in patients with any other TNM category. PMID- 6289014 TI - Common proviral integration sites in C57BL mouse lymphomas induced by radiation leukemia virus and absence of novel virus-related sequences in radiogenic lymphoma DNA. AB - Three C57BL/Ka mice were inoculated with RadLV/VL3, a thymotropic and leukemogenic virus population released by the permanent BL/VL3 cell line, which was derived from a C57BL/Ka lymphoma induced by radiation leukemia virus (RadLV). The neoplastic thymus and bone marrow cells from these mice were grown until the cultures became permanently established, and their DNAs were examined for the presence of virus-related sequences by restriction enzyme analysis. All six cell lines displayed identical EcoRI and BamHI restriction fragments, not found in control C57BL/Ka DNA and accounting for the presence of more than one novel provirus. The primary and secondary tumors were thus clonal and, even though they were obtained from different animals, possessed identical integration sites. The BL/VL3 cell line also displayed the clonal appearance of novel proviral sequences, partly identical, with respect to location and BamHI restriction pattern, to those found in the RadL/VL3-induced tumor cell lines. Neither radiation-induced tumors, nor cloned cell lines derived therefrom, whether producing leukemogenic virus (BL/RL12-P) or not (BL/RL12-NP), displayed the presence of novel virus-related sequences when compared with control tissues. Our results strongly suggest not only that, as described in the case of avian leukosis virus-induced tumors of the chicken, RadLV-induced leukemogenesis might be a consequence of cellular gene activation by promotor insertion, but also that more than one integration site might be involved. Radiation-induced tumorigenesis might be initiated by a comparable mechanism, not requiring the participation of a virus. PMID- 6289015 TI - Inhibitory effect of leukemic plasma on periodate-induced lymphocyte transformation. AB - Plasma from nine out of 18 patients with untreated acute lymphoblastic leukemia (ALL) depressed the transformation of normal blood lymphocytes induced by sodium periodate (NaIO4) as judged by reduction of blast cell formation and [3H]thymidine and [3H]uridine incorporation into DNA and RNA respectively. The depressed mitogen responsiveness of lymphocytes cultured in the presence of leukemic plasma was due to the presence of inhibitory factor(s) present in the plasma rather than the absence of components present in normal plasma. The inhibitory effect of leukemic plasma on periodate-induced cell stimulation indicated that the leukemic plasma inhibitory factor(s) exert their action very rapidly and directly on the cell. Transfer of the inhibitory factor(s) from the leukemic plasma to the cultured cells was supported by the finding that the depressive action of leukemic plasma on lectin and non-lectin mitogen-induced transformation of lymphocytes was reduced if the leukemic plasma was preincubated with either resting or mitogen-treated lymphocytes prior to testing with lymphocytes not previously exposed to leukemic plasma or mitogen. PMID- 6289016 TI - Limits of usefulness of electrophysiological methods for estimating dendritic length in neurones. AB - The present report deals with methodological problems concerning the electrophysiological analysis of multipolar neurones. In a nerve cell in which the dendrites may be considered as being equivalent to a single cable, the electrotonic length of the equivalent cable may be determined by analyzing the potential response to a step of current injected into the soma (Rall, 1969). In the present model study it is shown that this kind of analysis is unsuitable for discriminating between neurones with different dendritic lengths in excess of about 1.5-2.0 length constants. PMID- 6289017 TI - Malignant nonfunctioning paraganglioma of the retroperitoneum producing renovascular hypertension. AB - Malignant nonchromaffin paraganglioma (chemodectoma) is an unusual tumor arising from chemoreceptor cells derived from the neural crest. Although these tumors generally occur in the head and neck, where the term "carotid body tumor" applies, more than 20 cases of retroperitoneal origin have been described. This case report presents the clinical, radiographic, and pathologic features of a chemodectoma arising in the retroperitoneal area that produced severe hypertension by unilateral real vascular compression. The patient underwent exploratory laparotomy, and a left nephrectomy was done. Follow-up examination 1 year after surgery disclosed no clinical evidence of disease, and he had normal blood pressures without medication. PMID- 6289018 TI - Cardiac electrophysiology and its contribution to cardiology. PMID- 6289020 TI - [Etiologic diagnosis of generalized lymphadenopathy (case report of cytomegalovirus mononucleosis)]. PMID- 6289019 TI - Activation of contraction in cardiac muscle. AB - This paper begins with consideration of a qualitative model of excitation contraction coupling in cardiac muscle which is based to a large extent on experimental work carried out on isolated preparations of cardiac muscle by Earl Wood during sabbatical years in Europe in 1966-67 and 1972-73. It goes on to consider newer evidence about excitation-contraction coupling that has been obtained from studies of "skinned" fiber preparations (that is, muscles in which the cell membranes have been disrupted or are absent) and from studied of isolated preparations in which the photoprotein aequorin was used to monitor changes in sarcoplasmic Ca2+ concentration. The results obtained from these two kinds of study are then combined in a qualitative way to illustrate current ideas about the mode of action of inotropic interventions and to provide possible explanations of the Frank-Starling effect as it appears in isolated preparations of cardiac muscle. PMID- 6289021 TI - [Appearance of histiocytoma at the site of surgery for spinocellular carcinoma and skin transplant]. PMID- 6289022 TI - [Basal excretion and excretion after PTH phosphate and AMPc administration in non advanced chronic renal failure ]. PMID- 6289023 TI - [Urinary cAMP excretion after glucagon administration to patients with diseases of the liver and bile ducts ]. PMID- 6289024 TI - [Pseudoepileptic crises symptomatic of insulinoma]. PMID- 6289025 TI - [Cryoglobulinemic purpura as presentation of hepatocarcinoma]. PMID- 6289026 TI - Naltrexone, opiate addiction, and endorphins. PMID- 6289027 TI - Pharmacokinetics of ceftizoxime in patients undergoing hemodialysis. AB - Serum pharmacokinetics of 1 g ceftizoxime were studied in eight patients undergoing chronic haemodialysis. Arterial blood samples were collected at 3, 60, 120, 180 and 240 minutes and before the next dialysis 2-3 days after injection of ceftizoxime. Three minutes after injection the serum levels were 122 +/- 8.4 microgram/ml and 19.4 +/- 1.7 microgram/ml at the end of the dialysis. Mean elimination half-life under haemodialysis was 2.1 hours. Serum concentrations ranged from 4.9 to 13 microgram/ml two to three days after the haemodialysis. PMID- 6289028 TI - Naloxone-induced enhancement of carbon dioxide stimulated respiration. AB - In unanesthetized rats, naloxone (5 mg/kg, s.c.) produced an increase in both respiratory frequency and tidal volume as compared to saline administered animals. Maximal respiratory stimulation was observed within 5 minutes after naloxone injection and duration of the response was greater than 30 minutes. Exposure to different atmospheres of carbon dioxide potentiated the increase in ventilation in a step-wise manner as the carbon dioxide concentration was increased. Pretreatment with low doses of morphine sulfate (2 mg/kg daily for 2 days) or naloxone HCl (5 mg/kg daily for 5 days) enhanced respiratory stimulation induced by naloxone. It was concluded that naloxone increases the sensitivity of central ventilatory response to carbon dioxide as a result of displacement of endogenous endorphins from central opioid receptors. PMID- 6289029 TI - Corticosteroid, catecholamine and glucose plasma levels in rabbits after repeated exposure to a novel environment or administration of (1-24) ACTH or insulin. AB - The responsiveness of the adrenal cortex and the sympatho-adrenal-medullary system to stress factors and administration of (1-24) ACTH and insulin was studied in adult rabbits. In comparison to untreated animals, exposure to a novel environment for 10 min followed by artery puncture on 6 consecutive days elicited a moderate increase of corticosteroid (C), norepinephrine (NE) and epinephrine (E) plasma levels. Intramuscular injection of 50 micrograms/kg body weight (1-24) ACTH increased C, NE and E plasma levels. Saline injection resulted in elevated NE levels; C, E and glucose remained unchanged. After injection of 1.0 IU/kg body weight insulin C levels were higher than those found after exposure to a novel environment for 10 min followed by artery puncture; similarly, NE and E increased. In accordance with results obtained in the rat or mouse the sympatho adrenal-medullary system in the rabbit is stimulated by stress factors such as handling, artery puncture or injection of (1-24) ACTH or insulin. In contrast the adrenal cortex can be stimulated only to a certain extent by these manipulations. An increased activation of adrenal cortex cells occurs only after insulin, a maximum stimulation only after (1-24) ACTH administration. PMID- 6289031 TI - Exposure to 4-aminopyridine prevents depressant effects of opiates on sensory evoked dorsal-horn network responses in spinal cord cultures. AB - The depressant effects of morphine (0.1-1 microM) on sensory-evoked dorsal-horn network responses in explants of mouse spinal cord with attached dorsal root ganglia (DRGs) were rapidly restored after addition of 4-aminopyridine (4-AP; 0.1 mM) and major components of these cord responses were stably maintained in the presence of the opiate. Moreover, prior exposure of cord-DRG explants to 0.1 mM 4 AP prevented the depressant effects of 0.1 microM morphine on DRG-evoked dorsal horn responses, and the effects of 1-10 microM morphine were at least partly antagonized. Increased Ca++ levels (5 microM) attenuated the depression of dorsal horn responses by 1-10 micro M morphine and these effects of Ca++ were greatly enhanced in the presence of 4-AP--in some cultures, concentrations of morphine as high as 100 micro M were strongly antagonized during test periods up to 2 hours. Receptor assays showed that 0.1 mM 4-AP +/- 5 mM Ca++ had no effect on stereospecific opiate binding, indicating that the antagonist actions of these agents in our cultures do not occur at the level of the opiate receptor. The relevance of our in vitro studies of 4-AP antagonism of opiate-depressant effects on sensory-evoked dorsal-horn network responses for analyses of problems in opiate analgesia has been strengthened by a recent report demonstrating that 4-AP does, in fact, reverse morphine analgesia in rats, as determined by tail flick tests. PMID- 6289030 TI - Naloxone blocks morphine enhancement of kainic acid neurotoxicity. AB - Kainic acid (KA) is a potent convulsant which, when administered subcutaneously, induces sustained limbic seizures and a pattern of limbic brain damage that is thought to be seizure-mediated. Diazepam suppresses and morphine enhances both the seizures and brain damage induced by KA. Here we show that morphine enhancement of KA neurotoxicity is blocked in a dose-dependent manner by subcutaneous pretreatment with naloxone. Theses and related findings support the hypothesis that morphine enhances the seizure-linked neurotoxicity of KA by an opiate specific action at certain limbic receptor sites where opiates suppress GABAergic activity, thereby lowering the threshold for propagation of seizure activity in limbic circuits. PMID- 6289032 TI - A relationship between adenosine 3'-5'-cyclic monophosphate levels and deoxyribonucleic acid synthesis in Euglena. AB - Using the binding protein method we found that cAMP levels in normal, exponentially growing Euglena stay constant on per cell and protein basis. The level rises slightly when cells enter the stationary stage. Cells growing in low vitamin B12 medium show the same pattern during predeficiency growth. Upon becoming vitamin B12 deficient, the cAMP level decreases. Replenishment of these cells with the vitamin causes an immediate drop, followed by a sharp rise in cAMP. This is followed by resumption of DNA synthesis. The cAMP level drops and rises again when DNA duplication is completed and during the G2 period. The level of the cAMP drops again followed by resumption of cell division. the data suggest a relation exists between cAMP level, resumption and completion of DNA synthesis, and cell division. PMID- 6289033 TI - The ascorbate cyanide test and the detection of females heterozygous for glucose 6-phosphate dehydrogenase deficiency. PMID- 6289034 TI - [Effect of clinical factors on the results of radiotherapy of locally disseminated rectal cancer]. PMID- 6289035 TI - [Radionuclide studies in the evaluation of G.A. Ilizarov's method of treating chronic osteomyelitis accompanying bone damage]. PMID- 6289036 TI - [Experimental study of the mechanism of local radiation damage to the brain by means of penetration of 99mTc through the hematoencephalic barrier]. PMID- 6289037 TI - [Physico-tomographic preparation for the use of Bragg's peak in the irradiation of intracranial targets]. PMID- 6289038 TI - [Symptomology of non-invasive proliferative epithelial changes in the breast]. PMID- 6289039 TI - Urinary and plasma vitamin D3 metabolites in the nephrotic syndrome. AB - Using newly developed and established extraction, Lipidex-5000 chromatography, normal phase gradient HPLC, and ligand binding assay techniques we have directly measured plasma and urine levels of vitamin D3 and its metabolites in seven normal subjects and seven patients with nephrotic syndrome and normal renal function. Significant reductions in the plasma levels of vitamin D3, 24,25(OH)2D3, 25,26(OH)2D3, and 1,25(OH)2D3 were noted in all nephrotic patients. In conjunction with the plasma metabolite abnormalities, direct quantitative analysis of the urine in these patients revealed significant increases in nonconjugated 250HD3, 24,25(OH)2D3 and 1,25(OH)2D3. Significant correlations were noted between the plasma and/or urine metabolites and other mineral homeostatic parameters. The results indicate that the primary basis for the reductions in plasma vitamin D3 and its metabolites in the nephrotic syndrome is enhanced urinary excretion. The findings of normal serum ionized Ca and i-PTH levels in the patients with nephrosis suggest that reductions in bound and not free forms of vitamin D3 metabolites in plasma may occur in the initial stages of the nephrotic syndrome. PMID- 6289040 TI - Changes in phosphodiesterase activity of human subcutaneous adipose tissue during starvation. AB - The phosphodiesterase (PDE) activity, the basal rate of lipolysis and the basal cyclic AMP level in adipose tissue were determined in hypogastric and gluteal specimens obtained from 14 obese healthy subjects before and after one week of starvation. During starvation there was a significant increase in both the tissue level of cyclic AMP and the rate of lipolysis, whereas the apparent values of Vmax of the low and high Km forms of PDE decreased significantly-i.e. by about 50 and 30 percent. respectively. The substrate concentration of cyclic AMP at Vmax of the low Km PDE corresponded to the tissue level of the nucleotide. During, but not before, starvation there was a positive correlation between Vmax of the low Km PDE and the cyclic AMP level (r = 0.7-0.8). The metabolism in the tissues from the two fat depots exhibited similar variations during starvation. The findings suggest that the low Km form of PDE is inhibited during starvation. This may be one factor responsible for the starvation-mediated increase in the cyclic AMP levels and the rate of lipolysis in adipose tissue. PMID- 6289041 TI - Preparation and identification of alpha- and beta-tropomyosins. PMID- 6289042 TI - Preparation of individual smooth muscle cells from the stomach of Bufo marinus. PMID- 6289043 TI - Purification of smooth muscle phosphatases. PMID- 6289044 TI - Purification of nonmuscle myosins. PMID- 6289045 TI - Characterization of tubulin and microtubule-associated protein interactions with guanine nucleotides and their nonhydrolyzable analogs. PMID- 6289046 TI - Electron paramagnetic resonance of contractile systems. PMID- 6289049 TI - Lactate dehydrogenase isoenzyme 1 (LDH-1) in athymic mice with xenografts of a human testicular germ cell tumor. AB - Lactate dehydrogenase (LDH) and LDH isoenzyme activities were determined in tumor tissue, tumor cystic fluid, and serum from athymic mice with transplants of a human testicular germ cell tumor. High activity of LDH-1 was found in tumor tissue and tumor cystic fluid. By histochemical staining LDH was found in all tumor cells. Most tumor cells had a faint staining reaction while some cells dispersed throughout the tumor had a strong staining reaction. The serum LDH-1 in athymic mice with tumor transplants correlated markedly with the tumor volume. The results indicate that serum LDH-1 was derived from the testicular germ cell tumor transplants. PMID- 6289047 TI - Changes in pools of acid-soluble phosphorus compounds induced by phosphorus starvation in Neurospora. PMID- 6289048 TI - Cloning and restriction mapping of the yeast URA2 gene coding for the carbamyl phosphate synthetase aspartate-transcarbamylase complex. AB - Two yeast DNA pools inserted in a hybrid Escherichia coli-yeast vector pFL1 were used to transform E. coli and yeast aspartate-transcarbamylase-less strains to prototrophy. From the first pool--a BamHI yeast DNA digest--a 6.4 kb BamHI fragment was recovered that gave good complementation of the E. coli auxotrophy but poor complementation of the yeast auxotrophy. From the second pool--a partial Sau3A yeast DNA digest--five independent plasmids complementing either E. coli, yeast, or both were recovered. Each of the five plasmids possessed sequences in common with the 6.4 kb BamHI fragment. One of these plasmids, which complemented the two URA2 activities in yeast and which produced a carbamyl-phosphate synthetase, aspartate-transcarbamylase complex sensitive to UTP feedback inhibition contained the full URA2 gene. A restriction map of the URA2 gene has been constructed and seven different consecutive segments have been recloned in pBR322 to measure their hybridization with URA2 messenger RNA, allowing us to estimate the limits of the gene. PMID- 6289050 TI - Adrenocortical steroids, bone mineral content and endometrial condition in post menopausal women. AB - In 23 post-menopausal women, serum levels of cortisol, unconjugated dehydroepiandrosterone (DHA), dehydroepiandrosterone sulphate (DHAS), testosterone, unconjugated and total oestrone and prolactin were measured before and during an ACTH test. Significant positive correlations were found between basal levels of DHA and DHAS; DHA and unconjugated oestrone; DHA and total oestrone; testosterone and total oestrone and between unconjugated and total oestrone. ACTH significantly raised the levels of the steroids but not of prolactin. Significant positive correlations were found between basal levels and ACTH induced increments in DHA; between basal DHAS and increments in DHA and between increments in DHA and DHAS. A significant negative correlation was found between basal levels and increments in cortisol. No significant correlations were found between other combinations of hormone basal levels and/or increments. Significant positive correlations were found between basal levels of DHAS and the DHA response to ACTH respectively, and trabecular bone mineral content of the distal forearm. A significant correlation was also found between bone mineral content and pre-cancerous/cancerous state of the uterine epithelium. The results are a further support to the concept of a link between adrenal androgens and bone mineral density, and do also indicate a relation to endometrial pathology. The lack of correlation between cortisol and other steroids indicate different regulatory mechanisms. Prolactin does not seem to be involved in the regulation of the adrenal androgen synthesis. PMID- 6289051 TI - Investigation of genetic interaction between simian rotavirus SA11 and reovirus. AB - Rotavirus and reovirus are physically and genetically very similar. Their genetic interaction would have important implications for understanding both the epidemiology of rotavirus and the biological basis of rotavirus pathogenicity. A permissive line of rhesus macaque kidney cells, MA104, was infected with the simian rotavirus SA11 and various temperature sensitive mutants of reovirus (type 3 groups A, B, C, D, G, type 1 M79, and type 2 122). The mixed cell infections were carried out at permissive temperature in the presence of absence of proteolytic enzymes, and the progeny were plated at a non-permissive temperature in mouse L cells, which are non-permissive to SA11. The progeny clones were labelled with 32P and analyzed by polyacrylamide gel electrophoresis. There was no evidence for genetic reassortment. Under this heavy selection pressure, such reassortment would have been apparent if it occurred with a frequency greater than 10-5 to 10-3. PMID- 6289052 TI - Cloning of the RHO gene of Escherichia coli. AB - The map position of the rho gene on E coil chromosome, as determined by phage P1 generalized transduction, was determined. The gene order is ilv-rho-cya. By Hind III endonuclease shotgun technique, the rho+ gene was cloned into a lambda-Charon 25 vector. The sizing of the restriction endonuclease generated DNA fragments by agarose gel electrophoresis, the heteroduplex analysis of cloned DNA molecules by electron microscope and the demonstration of the synthesis of rho protein by the cloned DNA shows that a functional piece of DNA, containing the entire rho gene of E coli has been cloned. PMID- 6289053 TI - Oxygen-derived free radicals and local control of striated muscle blood flow. PMID- 6289054 TI - Degradation of cowpox virus in HVJ (Sendai virus) carrier cells. PMID- 6289055 TI - Lack of detectable DNA damage in cells productively infected with human cytomegalovirus. PMID- 6289056 TI - [3':5'-Cyclonucleotidephosphodiesterase in Yersinia pestis EV]. PMID- 6289057 TI - [Effect of cyclic adenosine-3',5'-monophosphate, chloramphenicol and actinomycin D on Gluconobacter oxydans biosynthesis of extracellular levansaccharase]. AB - The biosynthesis of levansucrase by Gluconobacter oxydans was shown to be induced in media containing sorbitol or fructose. An addition of glucose at a concentration of 0.1% to the culture growing in a medium with sorbitol stimulated the biosynthesis of levansucrase, whereas glucose at a concentration of 0.5-0.6% inhibited the enzyme synthesis by 20-30%. Gluconic acid, a product of glucose metabolism, also repressed the synthesis of levansucrase, but to a lesser degree than glucose. Cyclic adenosine-3',5-'monophosphate eliminated the repression by glucose added to a medium with sorbitol. An addition of chloramphenicol to the culture growing in a medium with sorbitol showed that the induction of levansucrase required de novo protein synthesis. The inhibiting action of chloramphenicol increased with its concentration. An increase in the concentration of actinomycin D from 5 to 200 micrograms/ml inhibited the bacterial growth by 50%, but stimulated the biosynthesis of levansucrase by 40%. PMID- 6289058 TI - Comparison of ranitidine and cimetidine in duodenal ulcer healing. AB - A single-blind trial in 50 outpatients was aimed at comparing the effectiveness of ranitidine (150 mg twice a day) and cimetidine (200 mg three times a day and 400 mg at night) in the healing of endoscopically proven duodenal ulcer. Patients were reassessed after two weeks of therapy, and then after four weeks immediately before repeat endoscopy. If the ulcer had not healed, treatment was continued for a further two weeks when patient assessment and endoscopy were repeated. By four weeks, ulcers had healed in 72% of patients receiving ranitidine and in 64% of those receiving cimetidine (P = 0.76). At six weeks, the healing rate was 92% in both groups. Symptomatic improvement was similar with both drugs. It is concluded that ranitidine and cimetidine are equally effective in healing duodenal ulcer. PMID- 6289059 TI - [Enzymatic determination of phospholipids in blood]. PMID- 6289062 TI - In vitro stimulation of lymphocytes by different strains of cytomegalovirus. AB - The in vitro stimulation of lymphocytes from 15 healty adult donors by 5 different stains of CMV resulted in a positive stimulation ratio of lymphocytes from CMV seropositive donors with each of the 5 different strains. Differences in the absolute strength of the response was observed among the 5 strains for each individual, but we concluded that the use of antigen prepared with 1 strain of CMV is sufficient to obtain a positive or negative result. A maximum value for the in vitro lymphocyte stimulation test, however, will only be obtained using different antigen concentrations from several different CMV strains simultaneously. PMID- 6289060 TI - Epstein-Barr virus induced proteins V: comparison of EBV-specific polypeptides from different virus strains. AB - EBV-associated polypeptides induced in different Epstein-Barr Virus (EBV) producing cell lines by the tumor promotor TPA, and from purified EBV particles derived from the same lines were radioactivity labeled and analyzed by immunoprecipitation with human VCA+MA+ sera. In virus-producing cells no significant differences in the molecular weight of 35S-methionine-labeled EBV associated polypeptide patterns could be observed. The analysis 125I-labeled polypeptides from purified virus particles of four different strains revealed that, in addition to common polypeptides, individual EBV strains contain strain specific high molecular weight glycopolypeptides. These polypeptides, constituting part of the membrane antigen complex, are present in varying amounts. While P3HR-1 virus particles contain a major component of 250 000 and small amounts of 340 000 molecular weight polypeptides, Q IMR-WIL virus particles have more 340 00 than 240 000 molecular weight polypeptides. Furthermore, in B95 8 particles and in particles from an EBV strain isolated from an African green monkey (AGM-EBV) respectively, large amounts of 360 000 and 250 000 polypeptides could be observed. Since these glycopolypeptides carry strain-, subgroup- and group-specific antigenic determinants, also found in virus strains produced in human and marmoset cells, it should be further investigated whether these differences in molecular weight are virus-strain- or cell-specific. PMID- 6289061 TI - Animal studies on the efficacy of vaccination against recurrent herpes. AB - Guinea pigs, latently infected with herpes simplex virus type 2 (HSV 2), were immunized with an HSV 2 vaccine derived from extracts of infected cells and either formol inactivated or detergent treated. Although such postinfection vaccination could significantly increase the titers of neutralizing antibodies, it had no effect on the course of recurrent herpes in these animals. PMID- 6289063 TI - [Reproduction of Powassan and West Nile viruses in Aedes aegypti mosquitoes and their cell culture]. PMID- 6289065 TI - [Retrospective study of Wilms' tumor in Italy (1970-1979)]. PMID- 6289066 TI - Forskolin as an activator of cyclic AMP accumulation and lipolysis in rat adipocytes. AB - Forskolin increased cyclic AMP accumulation in isolated adipocytes and markedly potentiated the elevation of cyclic AMP due to isoproterenol. In adipocyte membranes, forskolin stimulated adenylate cyclase activity at concentrations of 0.1 microM or greater. Forskolin did not affect the EC50 for activation of adenylate cyclase but did increase the maximal effect of isoproterenol. Neither the soluble nor particulate low-Km cyclic AMP phosphodiesterase activity was affected by forskolin. Low concentrations of forskolin (0.1-1.0 microM), which significantly elevated cyclic AMP levels, did not increase lipolysis, whereas similar increases in cyclic AMP levels due to isoproterenol elevated lipolysis. Forskolin did not inhibit the activation of triacylglycerol lipase by cyclic AMP dependent protein kinase or the subsequent hydrolysis of triacylglycerol. Higher concentrations of forskolin (10-100 microM) did increase lipolysis. Both the increased cyclic AMP production and lipolysis due to forskolin were inhibited by the antilipolytic agents insulin and N6-(phenylisopropyl)adenosine. Hypothyroidism reduced the ability of forskolin to stimulate cyclic AMP production and lipolysis. These results indicate that forskolin increases cyclic AMP production in adipocytes through an activation of adenylate cyclase. Lipolysis is activated by forskolin but at higher concentrations of total cyclic AMP than for catecholamines. PMID- 6289067 TI - Distance geometry analysis of the benzodiazepine binding site. AB - The in vitro binding constants to brain benzodiazepine receptor are known for a variety of benzodiazepines and also for four other different classes of strongly binding compounds. The binding data for a total of 29 drugs selected from these 5 classes was used to deduce a possible binding site model consisting of 15 site points and only 5 adjustable energy parameters. Even though some of the chemical structures differed radically, it was possible to fit the experimental data with a root mean square deviation of 1.1 kcal/mole. Apparently 5 non-hydrogen atoms in each ligand can occupy corresponding points in the site, and thus constitute a possible benzodiazepine pharmacophore. PMID- 6289064 TI - Animal papillomaviruses. PMID- 6289069 TI - A fluorescence enhancement assay for cellular DNA damage. AB - A fluorescence procedure is described for quantitative measurement of DNA damage in mammalian cells. The technique is based upon the time-dependent partial alkaline unwinding of cellular DNA followed by determination of duplex:total DNA ratios with bisbenzamide, which has a differential molar fluorescence with single stranded and duplex DNA. The method is rapid, does not require radioactive labeling of DNA, and is sufficiently sensitive to detect damage induced with 100 rads of X-irradiation. This method is standardized with respect to the alkaline unwinding unit, Mn0, and the unwinding constant, beta. Results obtained with this new technique and with hydroxylapatite chromatography for physical separation of single- and double-stranded DNA were confirmatory. The utility of the technique was demonstrated by detection of dose-related damage with X-irradiation and a variety of antineoplastic agents in unlabeled murine leukemia cells. PMID- 6289068 TI - A possible mechanism for elevation of glucose-6-phosphatase activity in kidney and liver of fluoride-treated rats. AB - Glucose-6-phosphatase (EC 3.1.3.9) activity in kidney and liver was found to be markedly elevated 3 hr after a single large dose of fluoride (NaF, 35 mg/kg, i.p.). The increases in renal and hepatic glucose-6-phosphatase activity were completely suppressed by adrenalectomy. Moreover, the increments were markedly suppressed by injection of dibenamine as an alpha-adrenergic blocker or by injection of propranolol as a beta-adrenergic blocker. PMID- 6289070 TI - Characterization of variant neuroblastoma clones with missing or altered sodium channels. AB - Variant neuroblastoma cell clones were selected for resistance to the cytotoxic effects of neurotoxins that cause persistent activation of sodium channels. Three classes of variant clones were obtained: sodium channel-deficient clones, which have markedly reduced numbers of functional sodium channels; scorpion toxin resistant clones, which have sodium channels with an altered interaction with scorpion toxin; and parental-type clones, which have functional sodium channels similar to the ones from N18 cells but have other heritable alterations that confer toxin resistance. The frequency of conversion to all three variant phenotypes was enhanced by treatment with the missense mutagen N-methyl-N'-nitro N-nitrosoguanidine (MNNG), suggesting that all three variant phenotypes are the result of mutational events. Incorporation of [35S]methionine into the alpha subunit of the sodium channel (Mr = 270,000; pI = 5.8 +/- 0.2) was studied in normal and variant clones by affinity chromatography on wheat germ agglutinin/Sepharose followed by analysis of labeled polypeptides by 1- and 2 dimensional gel electrophoresis. Sodium channel-deficient clones do not synthesize the alpha-subunit of the sodium channel, suggesting that mutations in these clones block expression of the gene for this protein subunit. The scorpion toxin-resistant clone LV10 synthesizes an alpha-subunit which has a molecular weight and pI similar to those of the parental clones within the resolution of the methods used. PMID- 6289071 TI - Temperature dependence of the benzodiazepine-receptor interaction. AB - The temperature dependence of the interaction of three benzodiazepines with their receptors in rat brain membranes containing about 2 microM (endogenous) gamma aminobutyric acid was investigated. van't Hoff plots of the equilibrium dissociation constants were linear for [3H]diazepam and its N-desmethyl derivative (N-desmethyldiazepam, NDz), whereas for [3H]flunitrazepam a break between two linear portions occurred at about 10 degrees. Binding of diazepam, NDz, and flunitrazepam (for the latter, at temperature greater than 10 degrees) is enthalpy-driven (delta H approximately -40 to -50 kJ/mole with a negligible contribution from delta S. The latter result indicates that the interaction is not a simple hydrophobic association, but that it may be more complex in nature, possibly involving a conformational transition of the receptor-ligand complex. The activation energy for dissociation of the [3H]flunitrazepam- and [3H]diazepam receptor complexes is about 100 kJ/mole. Consequently, the complexes dissociate about 100 times faster on changing temperature from 0 degrees to 37 degrees. PMID- 6289072 TI - Three adenylate cyclase phenotypes in S49 lymphoma cells produced by mutations of one gene. PMID- 6289073 TI - Characterization of peripheral-type benzodiazepine binding sites in brain using [3H]Ro 5-4864. AB - The binding of [3H]Ro 5-4864 to the peripheral-type benzodiazepine binding site in brain is characterized. The binding is saturable, high-affinity (KD = 1.6 nM), and reversible. The comparison of [3H]Ro 5-4864 and [3H]diazepam binding sites reveals major differences which include the following. There are about one-fourth as many peripheral-type binding sites than central sites in brain. Peripheral sites are present in many extranervous tissues and have a brain regional distribution distinct from that of the central-type receptor. The [3H]Ro 5-4864 binding site also is apparently highly localized in the nuclear membrane in contrast to the central-type receptor, which is synaptosomal. gamma-Aminobutyric acid has no effect on [3H]Ro 5-8464 binding, again in contrast to its marked effect on [3H]diazepam binding. Various putative benzodiazepine receptor ligands, such as purines, beta-carbolines, and kynurenamines, are also inactive as inhibitors of [3H]Ro 5-4864 binding. Blocking the benzodiazepine receptor by photoaffinity labeling decreases [3H]diazepam binding by more than 80% and has no effect on [3H]Ro 5-4864 binding. These results indicate that the peripheral-type benzodiazepine binding site in brain is a separate entity whose physiological function is probably distinct from that of the central-type benzodiazepine receptor. PMID- 6289074 TI - Interaction of histamine with gastric mucosal cells. Effect of histamine agonists on binding and biological response. PMID- 6289075 TI - Adrenergic ligand liposolubility in membranes. Direct assessment in a beta adrenergic binding system. AB - The binding of [3H]dihydroalprenolol to adipocyte membranes may be displaced not only from beta-adrenergic receptors but also nonstereospecifically from other membrane compartments by beta-adrenergic and other adrenergic agents. The EC50 of this nonstereospecific displaceable binding is directly related to the liposolubility (n-octanol:water partition coefficient) of the displacing ligand. PMID- 6289076 TI - Storage of biogenic amines in intact blood platelets of man. Dependence on a proton gradient. AB - The actions of ionophores with different ion specificities and of thrombin on the release of 14C-labeled 5-hydroxytryptamine, [3H]noradrenaline, and endogenous ATP were measured in human platelets suspended in media with various K+ and Na+ concentrations. Besides thrombin, those ionophores [monensin, nigericin, and the combination of carbonylcyanide-p-trifluoromethoxyphenyl hydrazone (FCCP) with nonactin and/or valinomycin] which cause a rapid collapse of H+ gradients induced a fast and virtually total release of 14C-labeled 5-hydroxytryptamine and [3H]noradrenaline into the various media. FCCP alone, which causes an inversion of the membrane potential to inside negative values, induced a considerably slower amine release. Changes in the K+ and Na+ gradients did not lead to amine release, nor did interference with energy transduction by antimycin A with or without glycolysis inhibitors. Monensin and FCCP did not release ATP, whereas thrombin, added before or after incubation of platelets with FCCP and monensin, caused a marked liberation of the nucleotide. It is concluded that in intact human platelets (a) the intragranular storage of 5-hydroxytryptamine and noradrenaline mainly depends on the proton gradient across the granular membrane, and (b) ionophores causing a collapse of H+ gradients induce non-exocytotic release of 5-hydroxytryptamine and noradrenaline from intracellular storage granules. PMID- 6289078 TI - Organization of loach ribosomal genes (Misgurnus fossilis L.). AB - The organization of ribosomal genes in the genome and amplified nucleoli of loach (Misgurnus fossilis L.), a teleost fish, was studied. Ribosomal DNA (rDNA) was isolated by density gradient centrifugation, digested by EcoRI endonuclease and mapped with 18S and 28S rRNAs. The transcription unit of ribosomal genes in oocyte nucleoli was investigated by an electron-microscopic visualization technique. The repeat unit of loach rDNA was shown to be split by EcoRI into two fragments: a constant fragment (3 . 10(6) daltons) and heterogeneous one (11-13 . 10(6) daltons, major type, and 7-8 . 10(6) daltons, minor type). The average size of the transcription units was found to be 2.5 +/- 0.2 microm. The lengths of nontranscribed spacers varied from 2.4 to 2.9 microm; however, shorter ones of 1.2-1.4 microm were also noticed. The data indicate that loach rDNA is organized as tandem repeat units of two types: a major fraction of 20-25 kb (14-17 . 10(6) daltons) and a minor one 15-16 kb (10-11 . 10(6) daltons). The heterogeneous length of the repeat units was predetermined by the heterogeneity of nontranscribed spacers. PMID- 6289077 TI - Investigation of membrane structure using fluorescence quenching by spin-labels. A review of recent studies. AB - Fluorescence quenching is the loss of fluorescence intensity which is observed when a fluorescent molecule or group interacts with another molecule or group, called the quencher. By use of tryptophan residues of proteins, together with specific probe molecules, quenching can be applied to problems of biological and model membrane structure. Quenching interactions are short range (less than 50 A) so that structure on the scale of molecular dimensions can be examined. This review summarizes the recent applications of fluorescence quenching by spin (nitroxide)-labeled molecules to problems of membrane structure, including determination of the distance of membrane-bound molecules from the membrane surface, the strength of lipid-protein interactions and the strength of protein protein interactions within membranes. The unique advantages and the limitations of this powerful method are examined. PMID- 6289079 TI - Identification of recombinant plasmids containing DNA sequences derived from the 3' end of ovine thyroglobulin mRNA. AB - This paper describes the cloning in E. coli of several DNA copies of the 3' portion of ovine thyroglobulin mRNA. The cDNA clones were identified by in situ cloning hybridization to 32P-labelled thyroglobulin cDNA and by positive hybridization-translation assays. The longest thyroglobulin cDNA fragment cloned (psTg21A, 1670 bp), was shown by S1 nuclease mapping to be an unaltered copy of the thyroglobulin mRNA. psTg 21A overlapped with another cDNA fragment (psTg 11, 330 bp) containing the poly(dA)-tail and corresponding therefore to the 3' end of the mRNA. When aligned together the two clones represent more than 20% of the thyroglobulin mRNA length. Another cDNA fragment (psTg 15, 350 bp) was identified as an internal portion of the thyroglobulin mRNA sequence. PMID- 6289080 TI - Glycoprotein and protein induced changes in liposome permeability. AB - By measuring the permeability across the lipid bilayer in the presence and absence of membrane-bound protein or glycoprotein it should be possible to get an impression concerning their ability to penetrate into the membrane bilayer. Proteins such as phospholipase A2 and acetylcholinesterase markedly increase the permeability in contrast to glycoproteins (ovomucoid and orosomucoid) and cytochrome C. The results may serve as an indication of the type of interaction between lipids and membrane components. PMID- 6289081 TI - Restriction patterns of adult chicken globin genes at early and late stages of embryonic development. AB - Globin mRNA isolated from anemic chicken was transcribed into cDNA and integrated into the Pst I cleavage site of plasmid pBR 322. After cloning in E. coli strain HB 101 and colony hybridization with 125I-labelled globin mRNA the plasmids of individual clones were characterized by hybrid arrested cell-free translation. Thus we could isolate clones containing alpha or beta globin chain nucleotide sequences. DNA was isolated from chicken blastoderms incubated for 18-20 h and from 11 d chicken embryos. A comparison of the restriction maps of the DNA from the two developmental stages with labelled nick translated plasmids and labelled cDNA did not indicate any globin gene rearrangements between these two stages of embryonic development. We conclude, that the adult chicken globin genes show a constant genomic organization during embryonic development. However, the restriction patterns of the globin gene family of the chicken strain investigated revealed some differences after 2 generations of propagation. PMID- 6289082 TI - [Synchronous (synphaseous) additive models for mitochondrial ATP synthetase complex and for other energy transducing systems of cells (a hypothesis)]. AB - There exist some energy transducing enzymes containing immobile molecules of substrates which are not exchangeable during protein functioning. According to the proposed models the immobile substrates are localized at the "idle" (or "partial" or "imitational") catalytic sites, which differ from normal ("working") active sites of enzymes. Only some steps of a complete reaction sequence which take place at the "working" sites are carried out at the "idle" sites. On the other hand, cyclic conversion of the immobile substrate at an "idle" catalytic site may include some steps which are absent in the "working" site cycle. The occurrence of identical steps on the "idle" and "working" catalytic sites allows to synchronize their action through conformational interconversions of tightly packed and structurally related "idle" and "working" subunits of the enzyme. The presence of covaletly bound substrates or substrates localized in closed cavities of the "idle" sites allows to synchronize the action of many monomers containing such sites due to the absence of the rate-limiting step of simultaneous saturation of many catalytic sites by substrate molecules from solution, and due to the lack of substrate inhibition on the "idle" sites. The functions of the "idle" sites are miscellaneous e.g. in ion-transporting systems these sites are directly involved in ion translocation. In the actomyosin complex the "idle" sites imitate conformational alterations of "working" sites, thus allowing synchronous functioning of the polymeric structure. Variations in the number of the "idle" sites operating simultaneously with one "working" site allow to regulate some parameters of enzymatic processes, e.g. the stoichiometry (number of transported ions per ATP hydrolysed (or synthesised) or electron-transported, or hv-absorbed ones) for ion transported systems or the ratio (velocity of contraction to developed efforts) for the actomyosin complex. PMID- 6289084 TI - [Physical mapping of actinophage Streptomyces coelicolor A3(2). VI. The use of deletion mutants of actinophage phi C31 for construction of phage vectors]. AB - Nonessential region responsible for G function has been identified in theta C31 phage genome by means of deletion mutants. The mutant phenotype is expressed upon theta C31 phage propagation in Streptomyces albus G strains differing in functioning of restriction and modification systems. Based on their increased resistance to EDTA, deletions were located in theta C31 delta 10 and theta C31 delta 65 phage mutants. Data are presented on physical mapping of nonessention region of theta C31 phage. The total length of this region is 24.1% of the overall length of DNA molecules. The DNA segment of 19.1% of the whole genome contains overlapped deletions. Theta C31 actinophage is proposed to be used as a cloning vector for Streptomyces. Various deletion mutants obtained, with the capacity of about 3 thousands base pairs may serve as "insertion vectors". The presence of the stretched nonessential genome region allows to use theta C31 phage as a "replacement vector". Then, insertion of foreign DNA to replace the EcoRI--C fragment of theta C31 DNA of 6.4 x 10(3) base pairs is possible. The phages comprising hybrid molecules may be selected for G and Lyg phenotypes. PMID- 6289083 TI - [Complexes of histone H1 with supercoiled DNA]. AB - The action of DNA topoisomerase I on the complexes of supercoiled DNA (DNA I) with histone H1 and other histones was studied at different ionic conditions and histone/DNA ratios. In the presence of 0.15 M NaCl at histone H1/DNA ratios lower than 0.25 (w/w) the relaxation of DNA I is not inhibited. Raising of H1/DNA I ratio up to 0.7 is followed by significant inhibition of DNA I relaxation. At fixed H1/DNA I ratio maximal inhibition is detected at 0.25 M NaCl. At NaCl concentrations lower than 0.1 M and higher than 0.3 M increasing of DNA I relaxation in the presence of H1 was observed. Electron microscopic studies show that increase of ionic strength or H1/DNA I ratio causes more dense packing of DNA molecules in the H1.DNA complexes. Changes in the structure of complexes agree with the increase of DNA I relaxation inhibition in these conditions. DNA I relaxation inhibition by H1 is drastically decreased by iodination of tyrosine 72 residue in the globular part of H1 molecule. Individual core histones inhibit DNA I relaxation at much higher histone/DNA ratios and show different dependence of inhibition on ionic strength. The results are discussed in terms of the possible role of H1 in chromatin condensation-decondensation. PMID- 6289085 TI - [Thermosensitive vector derived from RP1 plasmid for detection of transposable elements]. AB - The involvement of the transposable DNA element of E. coli K12 chromosome in integrative recombination of RP1 plasmid was studied. Using temperature sensitive for replication plasmid RP1ts12--the derivative of RP1 which contains mutated transposon Tnl, it was shown that integration of RP1 into host chromosome and Hfr formation may occur according to a mechanism mediated by chromosome IS-elements. Plasmids that are desintegrated from the chromosome of these Hfrs contain discrete DNA segments (IS-elements) and possess elevated frequency of integration into chromosome of rec+ cells. The latter was used for selection of RP1ts12 recombinants carrying chromosome IS. For identification of IS involved in RP1 integration the number of independent RP1ts 12 recombinants was subjected to restriction and heteroduplex analysis. By analysing recombinants integrated into bacterial chromosome with frequency 5 X 10(-3), a new IS-element of E. coli K12 designated IS111 was discovered. IS111-element is about 1500bp of length, contains Smal, Pst1 and BamH1 restriction endonuclease sites and was found in the same position on the plasmid RP1 in two different orientations. IS-elements that have been revealed in a number of other RP1ts12 recombinants were preliminary identified as IS1-like elements. One recombinants plasmid was found to have an IS5-like elements. The activity of IS-elements inserted into RP1ts12 in recA dependent integrative recombination was estimated. From the data of absolute and relative RP1ts12 integration frequencies mediated by IS111, IS1- and IS5-like elements a conclusion was made about the absence of E. coli K12 chromosome IS elements in RP1 plasmid. The Hfr-formation and chromosomal gene transfer by recombinant plasmids RP1ts12: IS111 were studied. The possibility to use insertion RP1ts12 derivatives for the estimation of copies number, mapping and definition of orientation of IS-elements in bacterial chromosome and the possibilities for detection of transposable DNA elements using RP1ts12 in a wide range of gram-negative bacteria are discussed. PMID- 6289087 TI - Somatic cell hybridization between bovine leukemia virus-infected lymphocytes and murine plasma cell tumors: cell fusion studies with bovine cells. AB - Hybridization of peripheral blood lymphocytes from bovine leukemia virus-infected cows with murine myeloma cells resulted in the generation of hybrid cells secreting immunoglobulins composed of various combinations of heavy and light chains of both bovine and murine Ig origin. Some hybrid cells derived from the light-chain producer, but non-secretor murine myeloma NSI cell line, secreted IgM molecules composed of bovine mu-chain linked to bovine and/or murine light chains. Other hybridomas secreted mouse and bovine light-chain dimers and/or monomers, or failed to secrete any Ig polypeptide chain whatsoever. Immunoglobulins secreted by hybridomas obtained upon hybridization of bovine cells with the IgG-secreting murine myeloma P2X63 cell line, contained bovine mu chain in one of the seven hybridomas obtained, and bovine light chain in two of them. All the cell lines secreted murine light- and gamma-chains. PMID- 6289086 TI - [Reaction of endonuclease EcoRI with DNA of the plasmid ColEI]. AB - Interaction of restrictase EcoRI with supercoiled and open forms of ColEI DNA is studied in the range 0-55 degrees. At temperatures 25-37 degrees superhelical ColEI DNA is converted to linear form, the most complete cleavage of supercoiled molecules were observed at 25 degrees. This reaction is not followed by accumulation of intermediate open circular DNA forms. At 45-55 degrees open circular DNA forms with nicks at the restriction site are formed along with linear molecules. Similar effects were observed at 10 degrees, whereas at 0 degree only open circular DNA molecules are formed. PMID- 6289088 TI - Persistent viral infections of man. PMID- 6289089 TI - The biology of viral hepatitis. AB - Dramatic advances in the understanding of the pathogenesis, pathophysiology, prevention, and treatment of the major viral diseases of the liver have been made. Hepatitis B and A viruses have been identified, with specific diagnostic serologic assays commercially available for these infections. The diagnosis of non-A, non-B hepatitis is currently made by exclusion. Morphological alterations in viral hepatitis are similar, regardless of the etiologic agent. Chronic viral hepatitis may be associated with hepatitis B and non-A, non-B, but not with hepatitis A. Persistent infection with hepatitis B virus is associated with an increased incidence of primary hepatocellular carcinoma. Viruses similar to the hepatitis B virus cause the same spectrum of liver disease in certain animals. With the development of a vaccine against hepatitis B virus infection, it may be possible to prevent a large proportion of worldwide chronic liver disease, as well as primary hepatocellular carcinoma. PMID- 6289090 TI - Mutagenic activity of 4 active-principle forms of pharmaceutical drugs. Comparative study in the Salmonella typhimurium microsome test, and the HGPRT and Na+/K+ ATPase systems in cultured mammalian cells. AB - The nitroimidazole derivatives used in human therapy have shown a strong mutagenic activity in bacterial tests using Ames strains of Salmonella typhimurium. Our study compared the response of 4 products of this family on bacterial target cells as well as on mammalian target cells (Chinese hamster V79 cells). The strong positive response on TA100 was greatly reduced on the nitroreductase-deficient strain TA100 Frl. Furthermore, no mutagenic activity was found in V79 mammalian cells that we examined for ouabain and 6-thioguanine resistance. PMID- 6289091 TI - Multiplicity reactivation of alkylating agent damaged herpes simplex virus (type I) in human cells. AB - Herpes simplex virus type I (Strain KOS) is inactivated by treatment with MMS, MNNG and HN2 as determined by plaque assay on Vero cell monolayers, or by an infectious center assay with FS2 cells, human foreskin fibroblast line. At a given dose of MMS and MNNG, survival of the virus was significantly higher at a multiplicity of infection of 1.0 PFU/cell compared to 0.01 PFU/cell. These results indicate that HSV-1 infected human cells are capable of repairing chemically induced lesions by way of multiplicity reactivation. No evidence for multiplicity reactivation with HN2-treated virus could be obtained, however. PMID- 6289092 TI - Quantitative forward-mutation specificity of mono-functional alkylating agents, ICR-191, and aflatoxin B1 in mouse lymphoma cells. AB - We have analyzed forward-mutation specificity in S49 mouse T lymphoma cells. Our criteria of specificity were based upon relative mutabilities of a panel of 3 genetic markers: resistance to 6-thioguanine (6TGr), dibutyryl cAMP (bt2cAMPr), and ouabain (OUAr). We tested 2 monofunctional alkylating agents, ethyl methane sulfonate (EMS) and N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), and 2 heterocyclic compounds, ICR-191 and aflatoxin B1 (AFb1). AFB1 was activated with rat-liver microsomal S9 plus cofactors. Expression-time lags of each genetic marker ranged from 2 days of OUAr mutations to 6 and 8 days for bt2cAMPr and 6TGr cells, with stable induced mutant fractions thereafter. The relative activity of each agent for each marker was assessed on the basis of its mutagenic efficiency at equitoxic doses. Specificity differences between the agents were determined by taking ratios of mutagenic efficiencies (RME) for the 3 possible pairs of markers. From these quantitative correlations and other data we conclude that both MNNG and EMS induce ouabain resistance (and probably bt2cAMPr and 6TGr) by similar mechanisms, almost certainly base substitutions. In contrast, ICR-191 and AFB1 are respectively less than 2 and 3% as efficient as MNNG for OUAr mutant induction relative to the activity of each agent for 6TGr mutagenesis. We infer that ICR-191 and AFB1 very rarely cause base substitutions in S49 cells, but that their activities are consistent with production of deletions, insertions or chromosomal aberrations. S49 cells demonstrate an unusually high specificity of mutagenesis at the OUAr locus compared to several other rodent cell lines. Thus, this panel of markers in S49 cells can be used as a sensitive, reliable screening system for mutagen detection and to discriminate among major classes of mutagenic mechanisms. PMID- 6289093 TI - Action spectrum for the in vitro induction of simian virus 40 by ultraviolet radiation. AB - A line of simian virus 40-transformed hamster kidney cells was exposed to ultraviolet radiation at eleven different wavelengths in the region 238-302 nm. An action spectrum derived from the resulting exposure-response curves for the induction of simian virus 40 from these cells exhibits a broad peak in the region 260-270 nm suggesting DNA as the major chromophore for this response. This conclusion is consistent with results obtained by other investigators who have noted viral induction by a number of DNA-damaging agents. PMID- 6289094 TI - Mechanism for the differential induction of mutation by S9 activated benzo[a]pyrene employing either a glucose-6-phosphate-dependent NADPH regenerating system or an isocitrate-dependent system. AB - A significant difference in mutation frequency has been observed in CHO cells exposed to benzo[a]pyrene with alternative activation systems. Each system employed rat-liver S9 homogenate with one using isocitrate dehydrogenase to provide reduced NADP, while the other method uses glucose-6-phosphate dehydrogenase. Total aryl hydrocarbon hydroxylase (AHH) activity was greater for the isocitrate dependent system, however, this yielded a lower level of HGPRT mutants. It was ascertained that this reduced mutation frequency may result from sequestering of B[a]P substrate by crystals in the medium, possibly calcium phosphate, which decreased the effective substrate concentration. This sequestration enhances B[a]P internalization, which would explain the dichotomy between the AHH values and the mutation frequency data. The production of specific B[a]P metabolites was also examined by reverse phase HPLC quantitation of extracts of solutions in which the two activation systems were used. The levels of 7,8 dihydroxybenzo[a]pyrene produced by the glucose-6-phosphate protocol were consistently greater than with isocitrate. This may also be a contributing mechanism for elevating the mutation frequency with this procedure. These results demonstrate several interactions between test compound, cells, and metabolizing system which must be considered with in vitro activation systems. PMID- 6289095 TI - International Commission for Protection against Environmental Mutagens and Carcinogens. ICPEMC working paper 5/1: Perspectives in mutation epidemiology. 1. Incidence and prevalence of genetic disease (excluding chromosomal aberrations) in human populations. AB - Detailed knowledge of the birth frequency or the cumulative incidence over all ages of genetic diseases in human populations is a prerequisite for assessing the magnitude of possible genetic hazards caused by environmental mutagens. However, both theoretical and practical difficulties are involved in precisely measuring the total frequency of these diseases. Two sets of data from large-scale population studies, one from Northern Ireland and the other from British Columbia, are compared with each other and with the results from ad hoc surveys for individual monogenic disorders. With due allowance for differences in approach, examination indicates that the data from the large-scale population studies are inadequate. However, it could provide a crude estimate of the total frequency of genetic diseases and a fairly reliable estimate of the individual frequency of certain genetic disorders with early onset that are familiar and readily diagnosed. In addition to environmental mutagens, there are a number of factors associated with current human activity that may change the incidence of genetic diseases. In order to monitor the human population for environmental mutagens, the change in frequency of sporadic cases of those genetic diseases that arose from fresh mutation and that can be easily detected as early as possible should be followed closely. The mechanism of data collection currently being employed in some countries for childhood cancers, certain congenital malformations, and inborn errors of metabolism could be extended to include the so-called sentinel phenotypes. The rationale and feasibility of using retinoblastoma and Wilms' tumor (nephroblastoma) as examples of such population monitoring are described. PMID- 6289096 TI - Electromyographic study of diabetic and alcoholic polyneuropathic patients treated with gangliosides. AB - A double-blind, randomized electromyographic investigation was conducted of the effects of cerebral ganglioside treatment on patients suffering from diabetic or alcoholic polyneuropathy. Cerebral gangliosides (50 mg once a day) administered to 15 diabetic and to 15 alcoholic neuropathic patients for 40 days, facilitated the reappearance of sensory potentials and significantly increased the MAP amplitude in median, ulnar, and peroneal nerves. In relation to ganglioside treatment, there was no significant change in the conduction velocities or in the distal latencies of these nerves, nor was there a change in the duration of the MAPs. On the basis of these results, it is suggested that the cerebral gangliosides are capable of inducing an improvement in the excitability of nerve fibers and of facilitating the processes of reinnervation, probably by means of an enhancement of fiber sprouting. PMID- 6289097 TI - Dual type of facilitation at the neuromuscular junction of the frog. AB - End-plate potentials were recorded intracellularly at the frog neuromuscular junction. Facilitation produced by conditioning shocks applied to the motor nerve was studied in two different ways: using either low-frequency trains in Mg blocked preparations or using brief high-frequency trains followed by a test shock of variable latency in D-tubocurarine-blocked preparations. In both cases, two distinctly different types of motor end-plates were observed. The first type showed a low quantum content and a marked facilitation without depression, while the second type showed a high quantum content, a poor facilitation, and a marked depression. The physiological significance of these findings is discussed. PMID- 6289098 TI - Exercise effects on recovery of muscle acetylcholinesterase from reduced neuromuscular activity. AB - In order to investigate the effects of reduced and subsequently increased neuromuscular activity on muscle acetylcholinesterase (AChE), rats had one hindlimb immobilized with plaster casts for 4 weeks and were killed either at the end of immobilization (group I), after 4 weeks of resumed normal activity following cast removal (group R), or after 4 weeks of resumed activity supplemented with a daily treadmill-walking task (group E). Immobilization resulted in a decrease in adductor longus muscle weight to 66.4% of control; total muscle end-plate-associated AChE was decreased to 51.4%. Total muscle ACh hydrolysis was not significantly affected. Mild daily exercise during recovery increased total muscle end-plate AChE to control levels after 4 weeks, while in group R the corresponding level was significantly lower (84.4%). Decreased neuromuscular activity has different effects on end-plate AChE and non-end-plate AChE. Mild endurance-type overload during recovery from immobilization can accelerate recovery of end-plate AChE activity to normal. PMID- 6289099 TI - Altered supernormality in multiple sclerosis peripheral nerve. AB - Supernormality is a regular part of the recovery cycle of nerve. Supernormality was expressed in terms of amplitude, and in 20 control subjects the mean (S2 conditioned/S2 unconditioned) ratio of the median sensory nerve action potential amplitude recorded at the elbow measured 285 +/- 174 (range 119 to 783). A supramaximal conditioning shock was delivered 6 msec in advance of the test pulse. Seventeen (42.5%) of 40 patients with multiple sclerosis (MS) failed to demonstrate supernormality, the (S2 conditioned/S2 unconditioned) ratio being less than 100. This abnormality probably reflects a delay in recovery cycle and is additional evidence implicating peripheral nervous system involvement in multiple sclerosis. PMID- 6289100 TI - Dna restriction-enzyme analysis of herpes simplex virus isolates obtained from patients with encephalitis. PMID- 6289101 TI - Association between HLA-Bw44 and small cell carcinoma of the lung. PMID- 6289102 TI - Discordant cortisol response in pituitary disease. PMID- 6289104 TI - Viruses in aging and cancer: introductory remarks. PMID- 6289103 TI - Autocrine secretion of peptide growth factors by tumor cells. AB - Transforming growth factors (TGF) are growth-promoting polypeptides that cause phenotypic transformation and anchorage-independent growth of normal cells. They have been isolated from several human and animal carcinoma and sarcoma cells. One TGF is sarcoma growth factor (SGF) which is released by murine sarcoma virus transformed cells. Whereas the TGF interacts with epidermal growth factor (EGF) cell membrane receptors, it is not detectable in culture fluids from cells which contain high numbers of free EGF cell membrane receptors. The SGF acts as a tumor promoter in cell culture systems, and its effect on the transformed phenotype is blocked by retinoids (vitamin A and synthetic analogs). The production of TGF by transformed cells and the responses of normal cells to the addition of TGF to the culture medium raise the possibility that cells "autostimulate" their growth by releasing factors that rebind at the cell surface. The term "autocrine secretion" has been proposed for this type of situation in which a cell secretes a hormone like substance for which it has external cell membrane receptors. The autocrine concept may provide a partial explanation for some aspects of tumor cell progression. PMID- 6289106 TI - Cancer as a failure of normal differentiation. PMID- 6289105 TI - Cell cloning as a tool to analyze the immune response. AB - Long-term lines of cloned, immunocompetent, nontransformed lymphocytes have the potential to allow the direct examination of many of the key aspects of lymphocyte activation and regulation. We describe here techniques for the long term propagation of nontransformed human and mouse B-lymphocytes. These lines have the shared characteristics in that they bear membrane Ig, that their growth depends on the presence of appropriate growth factors, and that, when properly stimulated, they secrete Ig. We found that the human lines are negative for Epstein-Barr nuclear antigen and that they can be cloned by limiting dilution. A long-term line of human B-lymphocytes we prepared by limiting dilution cloning contained only lambda light chain-bearing cells. PMID- 6289107 TI - Receptor-mediated endocytosis and the structural organization of the cell membrane. PMID- 6289108 TI - Aging and cancer: cyclic 3', 5'-adenosine monophosphate and altered gene activity. PMID- 6289109 TI - Yeast genetics. PMID- 6289111 TI - Structural requirements for affinity and intrinsic activity at the opiate receptor defined in 4-phenylpiperidine and related series. PMID- 6289110 TI - Changes in gene order and gene expression. AB - Studies on changes in gene position in germ line and somatic cell chromosomes during evolution and differentiation have led biologists to abandon the static view of chromosomes as invariant linear arrays of hereditary information. Prokaryotic cells contain several classes of DNA insertion elements which move from place to place in the genome and mediate chromosome rearrangements. Similar elements exist in a wide variety of eukaryotic organisms (yeasts, insects, plants, and vertebrates). In addition, both reversible and irreversible changes of chromosome primary structure provide developmental controls on gene activity in bacteria, bacteriophages, yeasts, trypanosomes, and mammalian lymphocytes. At least five recombination mechanisms are known to catalyze chromosome changes: 1) general homologous, 2) site-specific reciprocal, 3) illegitimate, 4) DNA splicing, and 5) replicative. Various combinations of these mechanisms can explain many different chromosome rearrangements and changes in gene dosage. Changes in gene position can alter gene expression in many ways, some of which we understand (such as insertional mutation and inversion of coding and regulatory sequences) and some of which are still unexplained. The activities of DNA insertion elements and somatic rearrangement systems are subjects to controls at several levels by specific regulatory systems, natural selection, and connection to cell lineage. Despite the recent increase in knowledge about the biological importance of changes in gene order on chromosomes, there are far more questions than answers, particularly about the mechanisms that coordinate recombination events and cell division in higher organisms. PMID- 6289112 TI - Preclinical pharmacology of Lilly compound LY150720, a unique 4-phenylpiperidine analgesic. PMID- 6289113 TI - Interactions between tetrahydrocannabinol (THC) and morphine in rats. PMID- 6289114 TI - Interaction of Ca++ with normorphine and beta-endorphin on the guinea pig ileum. PMID- 6289115 TI - Localization of the reward-relevant opiate receptors. PMID- 6289116 TI - Evidence for a single opioid receptor type on the rat deferens. PMID- 6289117 TI - Autoradiographic localization of the phencyclidine/sigma "opiate" receptor in rat brain. PMID- 6289118 TI - Development of selective tolerance to particular types of opiate receptors. PMID- 6289120 TI - Evidence for the release of endogenous opiates by morphine. PMID- 6289119 TI - Methadone-induced endorphin dysfunction in addicts. PMID- 6289121 TI - Structure-activity relationships of oxygenated morphinans. III. An exploration of the effect of the aromatic oxygen and 6-keto group on antinociceptive activity, receptor affinity, and narcotic antagonism. PMID- 6289122 TI - Serotonin and cyclic AMP close single K+ channels in Aplysia sensory neurones. AB - We have identified a serotonin-sensitive K+ channel with novel properties. The channel is active at the testing potential; its gating is moderately affected by membrane potential and is not dependent on the activity of intracellular calcium ions. Application of serotonin to the cell body or intracellular injection of cyclic AMP causes prolonged and complete closure of the channel, thereby reducing the effective number of active channels in the membrane. The closure of the channel can account for the increases in the duration of the action potential, Ca2+ influx, and transmitter release which underlie behavioural sensitization, a simple form of learning. PMID- 6289123 TI - Localization of beta-adrenoreceptors in mammalian lung by light microscopic autoradiography. PMID- 6289124 TI - The primary structure and genetic organization of the bovine papillomavirus type 1 genome. AB - The complete nucleotide sequence of the doubt-stranded circular DNA of bovine papillomavirus type 1 (BPV-1) was determined. Analysis of this sequence in conjunction with known transcriptional data for the virus provides a basis of determining the organization of the papillomavirus genome. All the major open reading frames are located on the same DNA strand. The region transcribed in BPV transformed cells contains open frames in all three translational frames whereas the region transcribed in productively infected bovine fibropapillomas is characterized by two large open reading frames partitioned by a single translational stop codon. The localization of sequences diagnostic of promoters and polyadenylation sites suggests that splicing is involved in the biosynthesis of the viral mRNAs. A sequence comparison indicates the genome organization of the bovine papillomavirus and that of the members of the simian virus 40 polyomavirus subgroup to be distinct, suggesting that these two groups of viruses are evolutionarily unrelated. PMID- 6289125 TI - A virus mutant with an insertion of a copia-like transposable element. PMID- 6289126 TI - Key structures in transposition. PMID- 6289127 TI - Noradrenaline blocks accommodation of pyramidal cell discharge in the hippocampus. PMID- 6289128 TI - Retroviruses induce granulocyte-macrophage colony stimulating activity in fibroblasts. PMID- 6289129 TI - Differential expression of cellular oncogenes during pre- and postnatal development of the mouse. PMID- 6289130 TI - Transposable elements, hybrid incompatibility and speciation. PMID- 6289131 TI - Heterogeneous sensitivity of cultured dorsal root ganglion neurones to opioid peptides selective for mu- and delta-opiate receptors. PMID- 6289132 TI - GABA directly affects electrophysiological properties of pituitary pars intermedia cells. PMID- 6289133 TI - Serum antibodies that distinguish between the phospho- and dephospho-forms of a phosphoprotein. PMID- 6289134 TI - The effect of some antidepressants on prejunctional muscarinic receptors on the sympathetic nerves of the isolated rabbit ear artery. AB - The antimuscarinic activity of amitriptyline, desipramine, iprindole, mianserin and viloxazine on prejunctional sympathetic nerve endings were compared in the isolated rabbit ear artery. In the presence of cocaine (10 micro M) and yohimbine (1 micro M), amitriptyline (0.5-1 micro M), desipramine (1-3 micro M) and iprindole (5-10 micro M) produced parallel rightward shifts of the concentration response curve for the inhibitory effect of carbachol (CCh) on responses to electrical stimulation of the preparation at 3 Hz. Mianserin (3 micro M) produced some inhibition but altered the slope of the concentration-responses curve to CCh while viloxazine (less than or equal to 10 micro M) produced no inhibition. The depression of tritium efflux by CCh from arteries preincubated in 3H noradrenaline was inhibited significantly (P less than 0.05) by amitriptyline (0.1 micro M) and desipramine (1 micro M) and not by iprindole (17 micro M), mianserin (3 micro M) or viloxazine (10 micro M). Amitriptyline was 10-fold more active than desipramine and at least 30-fold more active than the other antidepressants as a muscarine receptor blocking drug in this preparation. Thus, mianserin, viloxazine and iprindole exhibit much weaker antimuscarinic activity relative to amitriptyline on prejunctional muscarine receptors on sympathetic nerve endings compared with that observed by others for excitatory muscarine receptors in sympathetic ganglia. The findings support an earlier suggest that these receptors differ. PMID- 6289135 TI - Noradrenergic neurotransmission in the brain of a convulsive mutant mouse, differences between the cerebral cortex and the brain stem. AB - The Quaking mouse is a genetically determined model of convulsive disorders. We investigated the modulation of noradrenergic neurotransmission through alpha 2 adrenoceptors in the occipital cortex and the brain stem of this mutant. The endogenous levels of noradrenaline were similar in the cerebral cortex of the Quaking mice and their corresponding controls, while a significant increase of endogenous noradrenaline was found in the brain stem of the mutants. The rate of disappearance of noradrenaline in the cerebral cortex and the brain stem after injection of FLA 63 was identical in control and Quaking mice. The calcium dependent electrically evoked overflow of 3H-noradrenaline from slices of occipital cortex was inhibited by clonidine and enhanced by yohimbine in Quaking as well as in normal mice. The negative feed-back mechanism mediated by presynaptic alpha 2-adrenoceptors operates to a similar extent in both strains of mice. In contrast to the occipital cortex, in the brain stem, the amount of neurotransmitter released by electrical stimulation was significantly increased in Quaking mice when compared with controls. However, in the brain stem, the negative feed-back regulation of noradrenaline release operates to a similar extent in both strains of mice. When the endogenous levels of MOPEG were determined in the brain stem, they were found to be significantly higher in the Quaking mice when compared to the controls. The results suggest that an increase in noradrenergic neurotransmission in the brain stem, rather than in the cerebral cortex, could contribute to the behavioural abnormalities exhibited by the Quaking mice. PMID- 6289136 TI - Acute or chronic changes of noradrenergic transmission do not affect the alpha adrenoceptor-mediated inhibition of 3H-serotonin release in the cerebral cortex. PMID- 6289137 TI - Influence of 22-day treatment on the anticonvulsant properties of cannabinoids. AB - Mice were given delta-9-tetrahydrocannabinol (delta-9-THC) cannabidiol (CBD) or phenytoin (PHT) daily for 22 days. Drug activity was measured weekly in three different anticonvulsant tests: the maximal electroshock threshold, the 60-Hz electroshock threshold and the 6-Hz-electroshock threshold. In order to correlate potential pharmacodynamic and pharmacokinetic changes resulting from repeated treatment, brain-drug concentrations were determined at each test time. The results from the delta-9-THC study indicate that, although tolerance developed in all three tests, there were no changes in the brain-drug concentration. For CBD the pharmacodynamics were strikingly different: an increase in sensitivity to the drug developed in two of the tests, tolerance in only one test. Here again, there were no changes in brain-drug concentrations. The results of the PHT study differed from both the cannabinoids, for tolerance developed in one test, an increase in sensitivity in one test, and the activity was unchanged in the third test. Again, the brain concentrations remained constant throughout. The results demonstrate that both tolerance and increased sensitivity can develop concomitantly with anticonvulsant effects of the cannabinoids and PHT, and that these modifications in drug activity appear to result from cellular or functional rather than dispositional changes. PMID- 6289138 TI - An increase in cardiac alpha 1-adrenoceptors following chronic clonidine treatment. AB - Chronic treatment (22 days) of rats with clonidine (0.5 mg/kg s.c. twice a day followed by 20 h of withdrawal) resulted in a significant increase in the specific [3H]WB4101 binding to ventricular and intraventricular septal alpha 1 adrenoceptors but no alteration of the atrial alpha 1-adrenoceptors. Scatchard analysis indicated that the increase in the [3H]WB4101 binding to the clonidine treated cardiac tissue was due to an enhancement of the alpha 1-adrenoceptor density since there was a significant increase in the Bmax value for the [3H]WB4101 binding to the treated ventricles without a change in the Kd value. The specific [3H]WB4101 binding to cardiac alpha 1-adrenoceptors was not altered by the acute (1 day) or 7 days treatment with clonidine. Chronic treatment with clonidine had no significant effect on the specific [3H](-)DHA binding to the atrial and ventricular beta-adrenoceptor. The noradrenaline (NA) concentrations in the clonidine-treated ventricles and intraventricular septae were decreased by 16-20%. These data provide biochemical evidence compatible with a significant reduction of sympathetic outflow to the ventricular myocardium by clonidine. PMID- 6289139 TI - A novel analysis of concentration-dependence of partial agonism Ring demethylation of bupranolol results in a high affinity partial agonist (K 105) for myocardial and tracheal beta-adrenoceptors. AB - 1. Ring-demethylation of the pure antagonist bupranolol results in a ligand (K 105) which induces conformational beta-adrenoceptor changes leading to partial agonistic effects in heart and trachea. However, these conformational receptor changes are not accompanied by changes in receptor affinity, because the affinity estimates for K 105 and bupranolol did not differ for a variety of myocardial tissues (including ventricular beta-adrenoceptors labelled with 3H-(-) propranolol] and trachea, not even for tracheal receptor subtypes. 2. For the analysis of the concentration-dependence of the blocking actions of a partial agonist a double log-plot was derived, which includes the classical Schild-plot as a special case. The plot is based on the statistical analysis of the action of partial agonists by Marano and Kaumann (1976). They defined a slope m for the weighted regression of equieffective concentrations of agonist in the absence and presence of a concentration [P] of partial agonist P. We derived the dependence of m on [P], which is suitably expressed as: log (1/m-1) = log [P]-log Kp. For the case of a single class of non-interacting receptors the slope of the double log-regression should be unity. Our plot has incorporated information from complete concentration-effect curves, instead of a single concentration-ratio as in the Schild-plot. Analysis of data of K 105 with the new plot (intrinsic activity greater than 0) and the Schild-plot (intrinsic activity = 0) yielded slopes near unity, consistent with simple competition. PMID- 6289140 TI - Analysis of the beta 1 and beta 2 adrenoceptor interactions of the partial agonist, clenbuterol (NAB365), in the rat jugular vein and atria. AB - The potent bronchodilator, clenbuterol, was compared to other beta adrenoceptor agonists with regard to affinity and efficacy for interaction with beta 1, and beta 2 adrenoceptors in the rat jugular vein and atria. Clenbuterol was a potent partial beta adrenoceptor agonist in both tissues based on the following observations: 1. Maximal relaxation of the jugular vein and increases in atrial rate to clenbuterol were less than maximal responses to other beta adrenoceptor agonists. 2. Clenbuterol antagonized responses to the stronger agonist, isoproterenol, in both tissues and 3. the equilibrium dissociation constant for clenbuterol approximated the ED50 concentration for vascular relaxation and increases in atrial rate, a characteristic of some, but not all, partial agonists. Relative to other beta adrenoceptor agonists, clenbuterol showed high affinity toward both beta 1 and beta 2 adrenoceptors and selectively toward beta 2 adrenoceptors. Equilibrium dissociation constants were 38 and 6.3 nM for beta 1 and beta 2 adrenoceptors, respectively. The high affinity of clenbuterol toward beta 1 and beta 2 adrenoceptors was coupled to a low relative efficacy of clenbuterol to activate either beta 1 or beta 2 adrenoceptors. Most beta 2 adrenoceptor agonists such as isoproterenol or salbutamol require approximately 1 3% adrenoceptor occupation for 40-50% relaxation of the jugular vein whereas clenbuterol required approximately 100% adrenoceptor occupation for a similar response. Thus, based on our analysis, the high agonist potency of clenbuterol results primarily from the high affinity toward beta adrenoceptors rather than efficient activation of the adrenoceptor as occurs with isoproterenol or salbutamol. PMID- 6289142 TI - Rational use of local anesthetic drugs. PMID- 6289141 TI - A study of the sensitivity of rat brain alpha 2-adrenoceptors during chronic antidepressant treatments. PMID- 6289143 TI - [Ratio between uptake and secretion of noradrenaline by nerve endings of the cerebral cortex during elaboration of conditioned reflexes in the rat]. PMID- 6289145 TI - Viruses and Glomerulonephritis. PMID- 6289144 TI - [Blocking of the Ca-dependent inward current in the somatic membrane of mollusk neurons by an elevated intracellular pH]. AB - The action of elevated intracellular pHi (pHi) on the transmembrane ionic currents in the somatic membrane was studied in intracellularly perfused nerve cells from Helix pomatia. Following a change in pHi from 7.3 to 9.0 the amplitude of potassium outward current recorded simultaneously with the calcium inward current was significantly reduced. This was accompanied by a shift of its I-V curve to more positive membrane potential values. In case of the calcium inward current blocking by external Cd2+ ions no reduction of the outward current was observed. Only a shift of its I-V curve along the potential axis remained. The calcium inward current was practically the same. It is suggested that the elevated pHi selectively blocks the Ca-dependent component of the potassium outward current. PMID- 6289146 TI - Automodulation of genes: a proposed mechanism for persisting effects of drugs and hormones in mammals. AB - We propose that hormone receptors are encoded by specially organized genetic elements that have the capacity to manipulate their own structure with enzymes in controlled ways. An automodulating element codes for a cell surface receptor protein. When this detector molecule is stimulated by its ligand (either by a natural hormone or a drug analog), a signal is transmitted back to the nucleus to activate an enzyme pathway which alters the structure of the genetic element. In particular, the DNA alteration changes the expression of the receptor on the cell, either qualitatively or quantitatively, and hence the sensitivity of the cell to future hormone stimulation. Thus, drugs can permanently change cell phenotype. It is suggested further that automodulation also occurs in germ-line cells allowing effects of a drug or hormone exposure to carry over to following generations as a genetic trait. PMID- 6289147 TI - An analysis of altered attention in monkeys exposed to delta-9 tetrahydrocannabinol during development. AB - Monkey infants born to mothers treated chronically with 2.4 mg/kg delta-9 tetrahydrocannabinol (THC) showed an alteration in visual attention characterized by a failure to limit the duration of attention when looking at novel stimuli. Five off-spring of treated mothers were compared to nine controls in a standardized situation in which they could view slides through a small peephole. THC offspring looked longer at slides early in a session and during the first session in which a given slide was seen. The time of appearance of the peak duration in a series of attention episodes also differed between groups. These alterations in visual attention could have an influence on perceptual and cognitive processes. PMID- 6289148 TI - 5'-nucleotidase in sciatic nerves from adult and young rats: recovery in myelin is lower than the recoveries of 2', 3'-cyclic nucleotide-3' -phosphohydrolase and carbonic anhydrase. PMID- 6289149 TI - Polyphosphoinositide mono- and diphosphoesterases of three subfractions of rat brain myelin. AB - Phosphomonoesterase and diesterase that cleave phosphatidylinositol-4-phosphate (diphosphoinositide, DPI) and phosphatidylinositol-4,5-bisphosphate (triphosphoinositide, TPI) were detected in three subfractions of purified rat brain myelin, and some properties of the enzymes were studied. Monoesterase activity was stimulated by KCl, maximally at a concentration of 25 mM, and inhibited at KCl concentrations above 50 mM. Addition of boiled pH 5 supernatant of rat brain homogenate doubled the enzymic activity; EDTA was inhibitory. The specific activities were nearly equal in the "low density", "medium density", and "heavy density" myelin fractions but about 30% lower than in whole brain homogenate. The monophosphatase could be solubilized by extraction with 0.2% Triton X-100. The phosphodiesterase activity was inhibited by EDTA and EGTA and not stimulated by KCl or pH 5 supernatant. Specific activities were nearly equal in whole brain and myelin but were by about 60 percent elevated in the "heavy density" over the "low density" myelin fractions. These results show that hydrolases operative in the fast turnover of the inositide phosphate groups are distributed over the entire myelin structure. PMID- 6289150 TI - Effects of phenylalanine and its deaminated metabolites on Na+,K+-ATPase activity in synaptosomes from rat brain. AB - The effects of phenylalanine (PHE) and its deaminated metabolites phenylpyruvate (PHP), phenyllactate (PHL) and phenylacetate (PHA) on sodium and potassium activated adenosinetriphosphatase (Na+,K+-ATPase) in synaptosomes from rat brain were investigated. At very low concentrations (5-10 microM). PHE, PHL and PHA inhibited the activity, while PHP stimulated the activity. At intermediate concentrations (50-100 microM), all compounds had no effect, but at higher (0.5 1.0 mM) concentrations they inhibited the enzyme activity. Thus all the compounds tested showed a biphasic effect on the enzyme activity. Hydroxylamine inhibited the Na+,K+-ATPase activity when present alone; simultaneous addition of hydroxylamine and PHE, however, eliminated the inhibitory effects of each other. Reversal of mutual inhibition also occurred in the presence of hydroxylamine and very low (5-10 microM) concentrations of PHL or PHA. The inhibitory effects of PHE at aLl concentrations, and of PHL or PHA at low concentrations, were also eliminated in the presence of EGTA. The data indicate that inhibition of brain membrane Na+,K+-ATPase by PHE and by low concentrations of PHL and PHA may involve metal ions, but that the inhibition by high concentrations of these metabolites must occur by a different mechanism. Since Na+,K+-ATPase plays a central role in neuronal function, and the presence of excess PHE and its deaminated metabolites occurs in brain tissue under conditions of experimentally induced hyperphenylalaninemia and genetic phenylketonuria, the neurologic impairment in experimental and genetic PKU may in part be related to the deleterious effects of these compounds on brain ATPase. PMID- 6289151 TI - Effects of corticosterone on brain cholinergic enzymes in chick embryos. AB - The effects of corticosterone on the cholinergic enzymes, choline acetyltransferase (ChAT) and acetylcholinesterase (AChE) were studied in the chick embryonic brain. Chick embryos received either 0.25, 0.5, or 1.0 microgram of corticosterone via the air sac daily for three days during either embryonic days 6 through 8 (E6-E8), of cerebral neurogenesis, or days 10 through 12 (E10 E12), a period of cerebellar neurogenesis. Enzyme activities were determined in cerebral hemispheres, optic lobes, cerebellum and remaining brain at 10, 15, and 20 days of incubation. In embryos treated from E6 to E8, ChAT activity was generally higher at day 10 in cerebral hemispheres and optic lobes (cerebellum was not determined) while AChE activity was not affected. At day 20 ChAT activity of treated chick embryos was lower in the cerebral hemispheres and optic lobes, but not in the cerebellum; AChE activity was higher in the cerebral hemispheres, lower in the optic lobes, and not changed in the cerebellum as compared to controls. However, in embryos treated from E10 to E12 both cerebellar ChAT and AChE activities were higher at day 15 in comparison to controls. These data show that the hormonal effects were most prominent only in the brain areas undergoing neurogenesis during the period of hormonal treatment. Since AChE activity is also present in nonneuronal cells, the observed alterations caused by corticosterone may reflect glial cell responses to the hormone. Whether the hormone affects the final number and/or maturation of cholinergic neurons and/or glial cells remain to be investigated. PMID- 6289152 TI - [Simple metastasis of hepatocellular carcinoma to the skull--a case report]. AB - The patient, a 53-year-old man, was admitted to the Department of Neurosurgery, Nagasaki University Hospital in August 1979, because of a painful tumor in the lt parieto-occipital region. Physical examination on admission revealed a fist sized tumor in the lt-parieto-occipital region. The liver edge was palpable at two finger breadths beneath the right costal margin, but the liver itself was not palpable as a mass. Liver function was almost good except for GOT and ChE. Plain craniography showed an osteolytic change of about 10 cm X 10 cm in size without marginal hyperostosis and spicular formation. Bilateral external carotid angiography demonstrated a hypervascular mass, which was fed by the bilateral superficial temporal arteries and occipital arteries. CT scan also demonstrated a fist-sized tumor originating from the diploe with destruction of the skull bone. Tumor was strongly enhanced. A radical removal of tumor was performed. Tumor was solid with profuse bleeding and adhered tightly to the dura and the skin. The postoperative course was uneventful and postoperative CT scan demonstrated complete removal of the tumor. Histological diagnosis was hepatocellular carcinoma and the skull tumor was considered to be metastatic. CT scan of the liver showed a circumscribed low density area with ring enhancement at the posterior ventral site. As no metastasis was found in other organ except the skull, so primary lesion was radically removed at the Second Department of Surgery. The incidence of metastasis to the bone in hepatocellular carcinoma in autopsy studies has been considered rarely approximately from 4 to 14%. Moreover, the common sites of skeletal metastasis are the vertebral column, the ribs, the long bones and so on. Metastases to the skull bone was exceptionally rare and all cases hitherto reported had multiple metastases in other organ(s) and no literature was found about the distant metastasis to the skull bone only. PMID- 6289153 TI - Complete cerebral ischaemia in the rat: an ultrastructural and stereological analysis of the distal stratum radiatum in the hippocampal CA-1 region. AB - An ultrastructural study and a stereological analysis of the distal stratum radiatum in the hippocampal CA-1 region of the rat were performed after 10 min of complete cerebral ischaemia followed by 10, 20 and 60 min of blood reflow periods. Post-ischaemic changes were mainly limited to the region of synaptic terminals which showed either clumping or dispersion of the synaptic vesicle pools and damage to synaptic membranes. Presynaptic terminals and astrocytes were swollen after 10 and 20 min of reflow, but this abated after 60 min. Mitochondria in neurons showed varying degrees of swelling, but in astrocytes their structure was normal. There were no changes in capillaries. After 20 and 60 min of blood reflow, disruption of cell membranes was observed, mainly in the vicinity of the synaptic terminals. The size of the extracellular space diminished by approximately 30% in all three ischaemic groups. The data show that synaptic terminals are a primary and early target in the development of postischaemic nerve cell damage. PMID- 6289155 TI - An adrenergic link in the hypothalamic pathways which mediates morphine- and beta endorphin-induced hyperthermia in the rat. AB - The mechanism underlying the hyperthermia induced by intrahypothalamic administration of either morphine or beta-endorphin has been investigated in conscious rats. Direct administration of morphine (1--8 micrograms in 1 microliter) or beta-endorphin (1--3 micrograms in 1 microliter) into the anterior hypothalamus caused hyperthermia in rats at the ambient temperature (8, 22 and 30 degrees C) studied. The hyperthermia in response to opiods was brought about by both increased metabolism and cutaneous vasoconstriction. This hyperthermia, unlike the hypothermia induced by intraventricular administration of opiods was not blocked by naloxone nor did tolerance develop to the response. However, the hypothermia induced by intrahypothalamic administration of opioids was greatly reduced by pretreatment with intrahypothalamic administration of either yohimbine (a blocking agent of alpha-adrenergic receptors) or DL-propranolol (a blocking agent of beta-adrenergic receptors) in the rat. These observations suggest that an adrenergic link occurs in the hypothalamic pathways which mediate morphine- and beta-endorphin-induced hyperthermia in the rat. PMID- 6289154 TI - Assessment of the potential agonistic and antagonistic properties of ketamine at opiate receptors in the guinea-pig ileum. AB - The anesthetic agent ketamine was studied for its ability to interact with opiate receptors in the longitudinal smooth muscle-myenteric plexus preparation of the guinea-pig ileum. The drug was found to possess agonistic, but not antagonistic, activity on opiate receptors. Naloxone antagonized the effect of ketamine although to a lesser degree than the antagonism exhibited toward morphine. In addition to the opiate action, ketamine also produced a depression of the contractile responses of the ileal smooth muscle to acetylcholine and histamine. The concentrations of ketamine that produced this non-specific depression were generally higher than those needed to demonstrate the opiate effect. However, some overlap in the concentration ranges for the two actions were observed. Although the nature of the opiate action of ketamine suggested that it interacts with opiate receptors or sub-types of these differently than does morphine, the non-specific action of the drug on the ileal smooth muscle precluded a definitive analysis of differences in the opiate receptor preferences of the drug. PMID- 6289156 TI - Block of sodium current in myelinated nerve fibre with enkephalins. AB - The effects of leu- and met-enkephalin were investigated on the node of Ranvier of isolated nerve fibers of frog under current and voltage clamp conditions. When added to the external solutions, enkephalins (1--5 mM) caused a slight decrease in peak Na+ and steady state K+ currents. The action potential was not significantly affected. When added to the internal medium (by diffusion from the two cut ends of the fibre), enkephalins (less than 5 mM) drastically reduced the peak Na+ current without significantly affecting the steady state K+ current. The block of Na+ current was greatly accelerated and enhanced by repetitive depolarizations. The sodium current slowly recovered after the end of pulsing. The extent of the block and rate of accumulation increased with increasing magnitude and frequency of the depolarizing pulses. Internal applications of enkephalins induced a blockade of the action potential when the fibre was stimulated at frequencies above 0.1 Hz. The results suggest that during depolarizations, enkephalin molecules plug the inner end of Na+ channels or immobilize the channel gates leaving the channels in a closed configuration, and remain in or near the pores for a long time after the end of depolarizations. Possible physiological significance and molecular mode of action of enkephalins on myelinated nerve fibres are discussed. PMID- 6289157 TI - Relationships between norepinephrine and cyclic nucleotides in brain and seizure activity. AB - To characterize further the roles of norepinephrine (NE) and cyclic nucleotides in seizure mechanisms, an examination was made of the effects of several drugs purported to depress noradrenergic influence in the CNS on pentylenetetrazol induced seizure activity and regulation of cyclic AMP levels in the cerebral cortex and hippocampus in mice. Depletion of brain stores of NE with reserpine or treatment of neonatal mice with 6-hydroxy-dopamine decreased seizure latency and/or threshold and diminished seizure-induced accumulation of cyclic AMP in brain. Propranolol, a beta-adrenergic receptor antagonist, and yohimbine, an alpha 2-adrenergic receptor antagonist, had effects qualitatively similar to reserpine and 6-hydroxy-dopamine, but phentolamine, a mixed alpha-adrenergic antagonist, increased seizure threshold and latency and did not reduce the accumulation of cyclic AMP. None of the drugs tested had any consistent effect on the regulation of cyclic GMP levels in brain during seizures. These data are consistent with the hypothesis that cyclic AMP in brain may be mediating an inhibitory influence of NE on seizure activity. PMID- 6289158 TI - Slowing of cortical somatosensory evoked activity by delta 9-tetrahydrocannabinol and dimethylheptylpyran in alpha-chloralose-anesthetized cats. AB - Somatosensory and visual evoked potentials in the classical afferent primary pathways in cats anesthetized with alpha-chloralose were studied in order to characterize the effects of delta 9-tetrahydrocannabinol (THC) and the synthetic analog, dimethylheptylpyran (DMHP) on central sensory processing. THC and DMHP slowed primary cortical responses to radial nerve or ventralis posterolateralis (VPL) stimulation in a dose dependent manner. THC did not alter VPL activity evoked by radial nerve stimulation. Amplitude of the primary somatic response was depressed at the 4 mg/kg dose. Responses of visual cortex and lateral geniculate nucleus, evoked by stimulation of optic chiasm, were unchanged at the doses studied. However, the nonmodality-specific response of the anterior marginal association cortex to stimulation of either modality was depressed by THC. Data suggest the association cortex which exhibits sensory convergence was most sensitive to the effects of THC. Findings were consistent with a thalamocortical site of action for THC. DMHP produced similar effects and was more potent. PMID- 6289159 TI - Benzodiazepine mechanisms and drinking in the water-deprived rat. AB - Benzodiazepines (diazepam, midazolam, flurazepam and chlordiazepoxide) produced a hyperdipsia in rats which were well-adapted to a daily 22 hr water-deprivation schedule. The hyperdipsia occurred as a result of extensions in the time-course of drinking without impairment in the efficiency of drinking (rate of water intake). At doses larger than those associated with hyperdipsia, the rate of water consumption was markedly impaired, so that any extension in the duration of drinking was offset by the impaired efficiency. As a result, a non-monotonic relationship between dose of benzodiazepine and volume of water intake could be generated. The convulsant benzodiazepine, Ro5-3663, produced a reduction in drinking, at sub-convulsant doses and without general motor interference. This hypodipsia was completely reversed by concurrent treatment with either diazepam or midazolam. The results are discussed in terms of possible behavioural and biochemical mechanisms which may underlie benzodiazepine-induced hyperdipsia. PMID- 6289160 TI - Cholecystokinin octapeptide, caerulein and caerulein analogues: effects on thermoregulation in the mouse. AB - Cholecystokinin octapeptide (CCK-8), caerulein and seven out of ten analogues of caerulein produced in mice, after subcutaneous administration, a dose-dependent drop in rectal temperature. For the hypothermic effect, i.e. a decrease in temperature by at least 1.5 degree C, ED50's were estimated with these peptides and with morphine, haloperidol and chlorpromazine. Caerulein was most potent, having an ED50 of 4.6 nmol/kg. With the aid of naloxone, atropine, phenytoin and desipramine the hypothermic effect of caerulein could be separated from that of morphine and haloperidol. These experiments, together with those on the relationship between chemical structure and effect, led to the conclusion that the thermoregulatory action of CCK-like peptides may be independent from other central actions, and vice versa. PMID- 6289161 TI - In vitro and in vivo inhibition by lithium of enkephalin binding to opiate receptors in rat brain. AB - The in vitro and in vivo effects of lithium ions on opiate receptor binding were studied in the cerebral cortex, the hippocampus and the basal ganglia of the rat. In vitro, lithium ions inhibited enkephalin binding to opiate receptors through a reduction in the number of binding sites, whereas the affinity was unchanged or only slightly decreased. In vivo, long term ingestion of lithium (3 weeks), during which the rats were maintained at a serum lithium level of approximately 1 mM, also inhibited enkephalin binding to rat neuronal membranes (P2-fractions) through a reduction of the number of opiate receptor binding sites, whereas the affinity was unchanged. No lithium could be detected in the suspension of neuronal membranes from the lithium-treated rats, and no difference in the concentration of endogenous opioid peptides was found between control rats and lithium-treated rats. The opiate receptors from control rats and lithium-treated rats did no display any difference in lithium sensitivity. This data suggest that administration of lithium to rats in small doses reduces opiate receptor binding of enkephalin. PMID- 6289162 TI - Effects of acute delta 9-THC administration on EEG and EEG power spectra in the rat. AB - This study was designed to determine the acute effects of delta 9-THC on the cortical EEG with the spectral analysis technique. Adult female Sprague-Dawley rats were implanted with chronic cortical and temporalis muscle electrodes. Intraperitoneally administered delta 9-THC (5 and 10 mg/kg) produced a reduction in peak-to-peak voltage of the desynchronized cortical EEG during wakefulness. Associated spectral power was reduced to about 50% of control during the first hour after injection of delta 9-THC and gradually returned toward the control value over an 8-hr period. Occurrences of delta 9-THC-induced high-voltage EEG bursts, overriding the reduced EEG tracing, were associated with an EEG spectral peak at 6 Hz. The first few slow-wave sleep (SWS) episodes appearing after delta 9-THC administration were associated with more slow-frequency waveforms and more slow-frequency spectral power than with control slow-wave sleep episodes. During control rapid eye movement (REM) sleep episodes, an EEG theta wave pattern, with an associated spectral peak at about 8 Hz, was characteristic. Conversely, the first few REM sleep episodes emerging after delta 9-THC administration contained overriding high-voltage bursts, the related power spectra of which had two peaks at about 7 and 11 Hz. PMID- 6289163 TI - New implantable continuous administration and bolus dose intracarotid drug delivery system for the treatment of malignant gliomas. AB - A totally implantable system for the continuous and bolus delivery of intra arterial chemotherapeutic agents to patients with malignant gliomas is described. The system utilizes an Infusaid pump (Infusaid Corp., Sharon, Massachusetts), which discharges the drug directly into the internal carotid artery and is percutaneously refillable. This system has been utilized experimentally in primates and in the treatment of six patients with malignant gliomas. It seems that this system can be utilized safely as an experimental technique in the treatment of malignant gliomas. PMID- 6289164 TI - Measurements of the volume and density of intracerebral tumors by CT following therapy. AB - For the interpretation of curative measures in patients with cerebral tumors CT is of increasing importance. The therapeutic effects can be demonstrated by close follow-up studies without any of the disadvantage of invasive neuroradiological methods. Our investigations of 125 patients with cerebral tumors are based on volume and density determinations. The CT studies of removed or inoperable tumors followed by radiation and/or cytostatic therapy prove that the best results follow a combination of both. In the present cases however, if CT proves postoperatively, at the end of radiation or at the beginning of the application of cytostatics that there is a residual mass, a complete remission cannot be obtained. PMID- 6289165 TI - Primary nasopharyngeal chemodectoma. PMID- 6289166 TI - The selectivity of ion channels in nerve and muscle. PMID- 6289167 TI - Postsynaptic factors controlling the shape of potentials at the squid giant synapse. AB - The roles of rectification and cable properties of the squid giant axon in determining the shape of synaptic potentials generated at the giant synapse were investigated. Excitatory postsynaptic potentials were recorded in response to selective stimulation of the main presynaptic axon at various temperatures. Excitatory postsynaptic potentials elicited at low temperatures (less than 18 degrees C) exhibited a marked after-hyperpolarization or undershoot, while those recorded at higher temperatures did not. The postsynaptic current, recorded under voltage clamp conditions, did not show an undershoot. Furthermore, intracellular injection of tetraethylammonium chloride, to block the voltage-dependent rise in potassium conductance, also eliminated the undershoot of the excitatory postsynaptic potential. These results indicate that the duration of synaptic potentials at the squid giant synapse is reduced by rectification due to a delayed rise in potassium conductance. Computer simulations of these synaptic potentials suggested that the effects of rectification will be more prominent in spherical (isopotential) cells than in cells with more complicated geometries. PMID- 6289168 TI - The effect of yohimbine on sodium and gating currents in frog Ranvier node membrane. AB - The indolalkylamine alkaloid yohimbine induced two phenomenologically-different types of sodium current (INa) inhibition in the voltage-clamped frog node of Ranvier, a tonic and a phasic ('use-dependent') block. The latter developed during a repetitive membrane stimulation with short (5 ms) depolarizing pulses at frequencies at 1 to 10 Hz. Unlike repetitive pulsing, a single-long lasting (1 s) depolarizing step did not produce a phasic block. Turning on a hyperpolarizing prepulse (50 ms to E = -123 mV) immediately before each test pulse produced a gradual unblocking of Na channels, while a depolarizing prepulse (to -86 mV) enhanced the phasic block. Yohimbine blocked the outward INa much more strongly than the inward ones. Reduction of external Na+ ions concentration from 112 to 55 mM caused a shift in the voltage-department of yohimbine block to more negative voltages, which coincided with the shift of INa reversal potential. Sodium current inhibition produced by yohimbine was accompanied by partial depression of the intramembrane charge movements ('ON-response'). Modification of Na channels by batrachotoxin made the Na channels resistant to both tonic and phasic blocking action of yohimbine. The features of the yohimbine-induced block suggest an interaction of the drug with open Na channels. The current-dependence of yohimbine block indicates an electrostatic interaction between Na+ ion and charged (protonated) form of yohimbine within the channel lumen and suggests the localization of the receptor at the inner mouth of the channel. Binding of yohimbine to the channel receptor promotes the inactivation of this channel. Comparison of the effects of yohimbine on NA and gating currents with those of local anesthetics leads us to suggest that these drugs share a common receptor. PMID- 6289169 TI - Presynaptic depolarization of unmyelinated primary afferent fibers in the spinal cord of the cat. AB - Low intensity (1-20 micro A) intraspinal stimulation produces in the sural nerve of the anesthetized cat short latency responses (3-4 ms) due to antidromic activation of fibers conducting in the A range (43-65 m/s). With higher stimulus intensities (up to 400 micro A) late responses (120-250 ms latency) may also be recorded. Simultaneous recording from two sites in the sural nerve shows that the peripheral processes of the fibers generating the late responses have a conduction velocity between 0.8-1.3 m/s. Collision between antidromic and orthodromic responses further indicates that these fibers have a peripheral threshold 20-25 times that of the A fibers. The late responses were largest when the intraspinal stimulating electrode was located in the dorsal horn, in the region corresponding to Laminae II and III of Rexed. The above observations suggest that the late responses are due to population responses of C fibers which are antidromically activated in the dorsal horn. The excitability of the C fiber terminals is increased by conditioning stimuli applied to other cutaneous afferents with a time course resembling that of the excitability increase of the A fibers on the same nerve. It is suggested that the effectiveness of synaptic transmission from C fibers to second order cells may be modulated presynaptically. In the decerebrate cat the antidromic responses of C fibers are reduced, but not abolished, by reversible spinalization produced by cooling or by sectioning the thoracic spinal cord. This suggests in addition that in the decerebrate preparation the presynaptic effectiveness of the C fiber (presumably nociceptive) input may be tonically decreased by supraspinal influences. PMID- 6289170 TI - Polarization of primary afferent terminals of lumbosacral cord elicited by the activity of spinal locomotor generator. AB - The changes of electrical polarization of primary afferent terminals in the lumbosacral spinal cord have been investigated on immobilized, decorticated and spinal cats during fictive locomotion. Fictive locomotion was spontaneous or provoked by stimulation of either dorsal root or dorsal funiculi in the lumbar segments. The activation of the locomotor generator and appearance of fictive locomotion were always associated with a sustained dorsal root hyperpolarization. On the background of this positivity periodic negative dorsal root potential oscillations appeared synchronously with efferent discharges in motor hind-limb nerves. These periodic waves of primary afferent depolarization occurred in phase in different ipsilateral lumbosacral segments. On the contralateral side the periodic changes in dorsal root potential were out of phase during fictive stepping and in phase during fictive galloping. The use of Wall's technique has shown that tonic and periodic changes in dorsal root potential reflect the changes occurring in polarization of central terminals of cutaneous and muscle (Ia and Ib) groups of afferent fibres. It is concluded that the level of electrical polarization of primary afferent terminals is determined directly by the activity of the spinal locomotor generator; activation of the generator is followed by hyperpolarization of primary afferent terminals. By so modulating the polarization of afferent terminals, the locomotor generator can perform tonic and phase-dependent selection of afferent information. PMID- 6289171 TI - Synaptic excitation of the second and third order auditory neurons in the avian brain stem. AB - Synaptic potentials were examined in the second- and third-order auditory neurons of nucleus magnocellularis and nucleus laminaris in the chick. Brain stems of mature chick embryos were explanted and maintained in vitro for 4 to 8 h. Field potentials, extracellular spike potentials and intracellular potentials evoked by 8th-nerve stimulation were examined. Eighth-nerve stimulation reliability elicited four identifiable field potentials which could be attributed to: (i) the afferent volley of the 8th-nerve axons, (ii) postsynaptic responses of n. magnocellularis neurons, and (iii) ipsilaterally and, (iv) contralaterally-evoked n. laminaris postsynaptic responses. Intracellular-recorded postsynaptic potentials were characterized by a rapid rise time and short duration. They were apparently monosynaptic with a synaptic delay of 0.4 ms. In each n. magnocellularis neuron the 'fast' excitatory postsynaptic potentials were composed of 1 to 3 all-or-none components. 'Slow' excitatory postsynaptic potentials were characterized by a longer latency, a longer duration and graded amplitude variation in proportion to the intensity of 8th-nerve stimulation. Both 'fast' and 'slow' excitatory postsynaptic potentials had similar reversal potentials. Since the 8th nerve makes monosynaptic connection with n. magnocellularis neurons, it is likely that at this synapse the 'fast' excitatory postsynaptic potentials were produced, while the 'slow' potential may be attributable to the convergence of many boutonal synapses of unknown origin. Intracellular injections of horseradish peroxidase into n. magnocellularis revealed that its efferents bifurcate below the nucleus and send one axon to the contralateral n. laminaris while the other axon forms a highly divergent projection to the ipsilateral laminar nucleus. The intracellular records obtained from n. laminaris are consistent with this anatomical finding in that graded excitatory postsynaptic potentials were elicited by 8th-nerve stimulation. PMID- 6289172 TI - Excitatory effect of acetylcholine on different types of neurons in the first somatosensory neocortex of the rat: laminar distribution and pharmacological characteristics. AB - In rats anaesthetized with either urethane, pentobarbital or fluothane the effects of acetylcholine, cholinergic agonists and antagonists (applied by iontophoresis) were studied on single cortical neurons of first somatosensory region. The laminar distribution of the neurons excited by acetylcholine was determined by the reconstruction of each electrode track based on a dye-deposit made at the last recording site. Neurons were identified using antidromic stimulation of the pyramidal tract, the ventrobasal thalamus and the corpus callosum. Neurons excited by acetylcholine could be segregated into two groups: one encompassing layer Vb and the upper part of layer VI, the other more deeply located at the limit between the cerebral cortex and the subjacent white matter. Neuronal responses to glutamate and nicotine, unlike those to actylcholine were evenly distributed in the cortex. Pyramidal tract neurons had corticothalamic neurons were frequently excited by acetylcholine and were shown to be located with the first group of acetylcholine sensitive neurons. Commissural neurons were rarely excited by acetylcholine and were not restricted to either group. The analysis of neuronal responses to acetylcholine and various agonists (carbachol, nicotine, acetyl-beta-methylcholine, carbamyl-beta-methylcholine, butyrylcholine) and antagonists (atropine, mecamylamine) revealed a prominent but not exclusive muscarine character. It is included (i) that cortical neurons of first somatosensory cortex which are excited by acetylcholine belong to two populations, one consisting, at least in part, of projection neurons (upper group) and the other of interneurons (lower group); (ii) that cortical acetylcholine receptors are of a 'mixed' type strongly weighted toward the muscarinic side. PMID- 6289173 TI - Properties of the receptive fields of frog retinal ganglion cells as revealed by their response to moving stimuli. AB - Quantitative mapping of the excitatory receptive fields of frog retinal ganglion cells revealed that the precise spatial position and locus of maximal activity within the excitatory receptive field were dependent upon the exact sequence with which the moving stimulus scanned the region of visual space in which the excitatory receptive field was embedded. A first-order asymmetry dependent upon the direction from which the moving stimulus entered the excitatory receptive field was noted. A second-order asymmetry, orthogonal to the first and sensitive to the direction in which a moving stimulus systematically traversed successive columns with in the excitatory receptive field was also described. These findings indicate that complicated interactions, both excitatory and inhibitory, occur between component parts of the receptive field in the frog retina. PMID- 6289174 TI - Are presynaptic dopamine autoreceptors and postsynaptic dopamine receptors in the rabbit caudate nucleus pharmacologically different? AB - Slices of the rabbit caudate nucleus were preincubated with [3H]dopamine or [3H]choline and then superfused and stimulated electrically. Apomorphine reduced the stimulation-evoked overflow of tritium over the same concentration range, independently of whether slices had been pre-incubated with [3H]dopamine or with [3H]choline. Each of three antagonists--molindone, sulpiride and metoclopramide- increased the evoked overflow of tritium over the same concentration range in experiments with [3H]dopamine and those with [3H]choline. For each antagonist, the pA2 values against apomorphine obtained in [3H]dopamine experiments and in [3H]choline experiments were very similar. This study is a functional in vitro approach to receptor characterization, as opposed to radioligand binding studies or in vivo investigations. The results show that the dopamine receptor agonist apomorphine and three antagonists are unable to distinguish between the presynaptic, release-inhibiting dopamine autoreceptors and those postsynaptic dopamine receptors which, when activated, depress the release of acetylcholine. Although there are certainly more dopamine receptors in the caudate nucleus, these two physiologically important groups seem to be closely related. PMID- 6289176 TI - The gadolinium ion: a potent blocker of calcium channels and catecholamine release from cultured chromaffin cells. PMID- 6289175 TI - A peptide-containing fraction of plasma from schizophrenic patients which binds to opiate receptors and induces hyper-reactivity in rats. AB - A serum fraction from schizophrenic patients has been investigated for its effect on opiate receptor sites and on behaviour in rats. Serum from schizophrenic patients was ultrafiltered and fractionated on DEAE-Sephadex A-25. The concentration of peptide material eluting under 0.1 M HCl (fraction I) was further purified on Sephadex-G10 and four major peaks were identified (fractions II to V). When injected intracerebroventricularly in rats, fraction II produced a characteristic behavioural syndrome, which included hyperactivity associated with hyperemotionality. The effects were long lasting, bouts of hyperemotionality accompanied by analgesia were recorded over a two-week period. The same fraction from control non-patients produced a transient and much reduced, but qualitatively similar response. There was evidence that fraction III was also active. An in vitro opiate receptor binding test showed that fraction II from schizophrenic patients inhibited [3H]naloxone binding. PMID- 6289177 TI - Parallel changes in ultrastructure and noradrenaline content of nerve terminals in rat vas deferens following transmitter release. AB - Transmural electrical stimulation and exposure to incubation media where some or all of the Na+ had been replaced with K+ were used to elicit transmitter release. Changes in noradrenaline content and ultrastructure of the nerve terminal varicosities in rat vas deferens were measured. Electrical stimulation in the presence of 4-aminopyridine had little effect, but high [K+] solutions caused a parallel reduction in noradrenaline content and the number of small dense-cored vesicles; large dense-cored vesicles showed no change, and small clear vesicles increased in number. In spite of a reduction in total vesicle number there was no evidence of expansion of the varicosity membrane. The parallel fall in noradrenaline content and in the number of small dense-cored vesicles suggests that the latter are the source of the released noradrenaline under the conditions of high [K+] stimulation we have used. PMID- 6289178 TI - Stereoselectivity of opiate antagonists in rat hippocampus and neocortex: responses to (+) and (-) isomers of naloxone. AB - The relative potencies of the (+) and (-) isomers of naloxone in antagonizing electrophysiological responses to D-alanine2-methionine enkephalinamide were compared in rat frontal cortex and hippocampus. In the in vitro hippocampus, the (-) isomer was found to be at least a 100 times more potent than the (+) isomer in antagonizing opiate-induced changes in field potentials. Similar stereoselectivity was observed in vivo in both frontal cortex and hippocampus in terms of the antagonism of enkephalin-induced changes in spontaneous cell firing. The direct effects of (+) and (-)-naloxone were examined as well. In hippocampus both in vivo and in vitro, no differential effect was observed, whereas in the neocortex (-)-naloxone was considerably more potent than the (+) isomer in eliciting depressions of spontaneous activity. These direct effects of naloxone in the cortex do not appear to be due to an antagonism of the effects of endogenously released opioids. These results demonstrate that the stereoselectivity of naloxone isomers in antagonizing electrophysiological responses to opiates in the cortex and hippocampus parallels that previously observed in other brain regions and in other tissues. In addition, they suggest that naloxone may have interactions with other unknown opiate (or possibly non opiate) receptors which are of physiological significance. PMID- 6289180 TI - Ammonium sulfate induced uncouplings of crayfish septate axons with and without increased junctional resistance. AB - Cytoplasmic pH (pH1) of crayfish lateral giant axons was monitored using antimony microelectrodes placed near septate junctions and variations of internal pH was induced by short applications of ammonium sulfate in the perfusing bath of the preparation. This treatment produced a rapid cell alkalinization followed, after wash, by acidification rebound. Simultaneously, two successive phases of uncoupling of the septate junction occurred; they had the same time course as those of their associated pH1 movements. Calculation of the electronic coupling parameters indicated that, during alkalinization, coupling was accompanied by an increased axonal membrane conductance (the intimate origin of which was beyond the scope of this study) and resulted from a shunt of the gap junctions; the resistance proper of the latter was unaffected; thus involvement of Ca2+ was ruled out and uncoupling was only an indirect consequence of the electrotonic junction's network configuration. In contrast, and as expected from previous investigations, the junctional membrane resistance was increased during the second phase of cytoplasmic acidification. Evidence that uncoupling can be brought about by a non-junctional membrane increased permeability raises questions about some of the criteria commonly used during investigations of electrotonic transmission. PMID- 6289179 TI - Electrotonic and dye coupling in hippocampal CA1 pyramidal cells in vitro. AB - The presence of electrotonic and dye coupling in region CA1 of the guinea-pig hippocampus was investigated in the in vitro hippocampal slice preparation. No electronic coupling potentials were observed in simultaneous recordings from 101 pairs of pyramidal cells. Also, no electrotonically-coupled short latency depolarizations were observed in more than 75 pyramidal cells in response to antidromic activation of the pyramidal cell population, either in normal bathing medium or in medium with lowered Ca2+ concentration and added Mn2+. When the fluorescent dye Lucifer Yellow was injected into pyramidal cell somas, spread of the dye to other cells (dye coupling) was often observed. Injection of Lucifer Yellow into the dendrites of these neurons resulted in many fewer cases of dye coupling. The failure to find electrophysiological evidence of electrotonic coupling among CA1 pyramidal cells suggests that such coupling is not a functionally important feature of this area of the CNS. The lack of electrophysiological evidence of coupling combined with the observation that the site of Lucifer Yellow injection influences the extent of dye coupling further suggests that at least part of the observed dye coupling may be artifactual. Electrotonic coupling may exist in a small percentage of hippocampal pyramidal cells. However, it is not clear that this small amount of coupling is either necessary or sufficient for the synchronization of neural activity as has been hypothesized to occur during epileptogenesis. PMID- 6289181 TI - Comparison of the effects of repeated electroconvulsive shock on alpha 2- and beta-adrenoceptors in different regions of rat brain. AB - The effects of ten, once-daily electroconvulsive shocks on alpha 2 and beta adrenoceptor binding were investigated in the rat cortex, hippocampus, hypothalamus and cerebellum. [3H]Clonidine and [3H]dihydroalprenolol were used as radioligands for alpha 2 and beta-adrenoceptors respectively. Twenty-four hours after the last shock, the density of beta-adrenoceptors was reduced in the cortex and hippocampus, but not in the hypothalamus or cerebellum. There was no change in the apparent affinity of the beta-receptors in any of the regions studied. Alpha 2-adrenoceptor density was reduced in all the regions studied (cortex, hippocampus, hypothalamus) with, again, no change in their apparent affinity. It is concluded that the effects of electro-convulsive shock on alpha 2 and beta adrenoceptors show regional variation; possible reasons for this variation are discussed. PMID- 6289182 TI - Differential distribution of immunoreactive angiotensin and angiotensin converting enzyme in rat brain. AB - To help resolve the controversy about the brain renin-angiotensin system, the distribution of immunoreactive angiotensin in the brains of male rats was analyzed using twelve different antibodies to angiotensin II, two of which had previously been reported to stain nerve fibers in the central nervous system. The distribution of angiotensin-converting enzyme immunoreactivity was also examined using an antibody to rabbit lung converting enzyme, and the distribution of this immunoreactivity was compared to that of immunoreactive angiotensin. Weak angiotensin-like immunoreactivity was found in cell bodies of the hypothalamic magnocellular nuclei of colchicine-treated rats and in nerve terminals of the median eminence, neurohypophysis, central nucleus of the amygdala, bed nucleus of the stria terminalis and various other sites in the brain and spinal cord of untreated rats. Staining could be demonstrated with only three antisera. Antigenic specificity was carefully studied in these antisera. Each was similar in that staining could be blocked with angiotensins I, II or III and tetradecapeptide renin substrate, although angiotensins II and III were most potent. Because of the relatively few angiotensin II antisera which could stain brain and because they are blockable with angiotensin I and tetradecapeptide renin substrate, the precise nature of immunoreactive angiotensin remains an open question. Intense converting enzyme-like activity was localized in endothelial cells of capillaries throughout the brain, in the subfornical organ and in the 'brush border' of choroidal epithelial cells in contact with cerebrospinal fluid. No activity was detected in neural tissue other than the subfornical organ and occasional weak activity in some ependymal elements elsewhere. These findings indicate that angiotensin and converting enzyme immunoreactivities are not co distributed and raises several questions regarding the nature of, and pathway for, formation of immunoreactive angiotensin in the brain. PMID- 6289183 TI - An ultrastructural study of neurons and non-neuronal cells in the myenteric plexus of the rabbit colon. AB - The myenteric plexus of the rabbit colon showed a degree of structural organization that was unusually high for the peripheral nervous system, providing a basis for the complex integrative activity which is known to occur. It resembled central nervous tissue in several respects: a wide range of neuron types was present; the proportion of glial cells to neurons was about 2:1; and there was a densely packed, avascular neuropil, not penetrated by connective tissue. Most neurons had at least one surface exposed to the extraganglionic space. Clear evidence was obtained for spontaneous neuronal degeneration. Three types of non-neuronal (glial) cells were observed: Type 1, which was most common, contained many 10 nm 'gliofilaments' and resembled enteric glial cells or astrocytes in the central nervous system; Type 2, composing about 5% of the glial cells, had few filaments; Type 3 was seen only rarely, had a small dark nucleus, little cytoplasm, may have been of extraganglionic origin and resembled microglia of the central nervous system. Fibroblast-like cells were also present in extraganglionic sites. Schwann cells could not be identified within the myenteric ganglia. PMID- 6289185 TI - Quantitative investigations of development of synaptic junctions in neocortex. PMID- 6289184 TI - The development and modifiability of the cerebral cortex. Overview. PMID- 6289186 TI - Normal and abnormal development of cerebral cortex in man. PMID- 6289187 TI - Peripheral neuropathy in multiple system atrophy with autonomic failure. AB - Two patients with multiple system atrophy, autonomic failure, and peripheral neuropathy are reported. EMG conduction study in both muscle and sural nerve histology in one patient documented the involvement of the neuromuscular system. Morphologic study of the biopsied nerve showed marked reduction of large myelinated fibers, whereas the unmyelinated axons were totally spared. The latter finding provides evidence that the syspathetic nervous system contributes few, if any, axons to the total population of unmyelinated fibers in the human sural nerve. Peripheral nerve damage may be common in the Shy-Drager syndrome. PMID- 6289188 TI - Fabry disease: detection of heterozygotes by examination of glycolipids in urinary sediment. AB - Fabry disease is an X-linked sphingolipid disorder that is manifest clinically as a disease of nerves, kidneys, and blood vessels. Precise identification of Fabry heterozygotes is essential for genetic counseling. Heterozygote detection by enzyme assay does not consistently distinguish them from unaffected females. We describe a method for Fabry heterozygote detection, based on quantitation of urinary sediment glycolipids by high-performance liquid chromatography. In specimens from 12 Fabry heterozygotes, the total glycolipid fraction was increased (10 to 100-fold) and trihexosyl ceramide (CTH) was 2- to 70-fold times normal. Digalactosyl ceramide (Digal-Cer), which is normally present in trace amounts in urine, was also increased. The ratio of CTH and Digal-Cer to hydroxy fatty acid glucosyl ceramide was increased and seemed to be characteristic of Fabry disease. This method provides rapid and accurate detection of Fabry heterozygotes. PMID- 6289189 TI - Asymptomatic sensorimotor polyneuropathy in workers exposed to elemental mercury. AB - Neurologic and electrodiagnostic evaluations and urine mercury level determinations were performed on 138 chlor-alkali plant workers, some of whom were chronically exposed to inorganic mercury vapor. Eighteen subjects had a mild polyneuropathy on clinical examination. These subjects had significantly (p less than 0.05) elevated urine mercury indexes, reduced sensation on quantitative testing, prolonged distal latencies with reduced sensory evoked response amplitudes, and increased likelihood of abnormal needle electromyography compared with the remaining 120 subjects. Similar results were found for subgroups matched by sex and age. We conclude that elemental mercury exposure is associated with a sensorimotor polyneuropathy of the axonal type; the degree of neurologic impairment appears related to the magnitude of exposure. PMID- 6289190 TI - [Angiodysplasias of the limbs]. PMID- 6289191 TI - [Surgical treatment of endocrine tumors of the pancreas]. AB - Following a brief review of the literature dealing with the rarity of this disease, and the diagnostic progress made to date, the surgical techniques applicable to the various locations of the neoplasia are described. Personal experience with 8 insuloma patients is reported. PMID- 6289192 TI - [Current role of angiography in the study of liver diseases]. AB - The Authors remember that also nowadays the angiographic studies for hepatic neoplasms is still interesting. These methods must be used after other less invasive screening methods, such as scintigraphy, ecotomography and computed tomography. Angiographic studies are essential in presurgical planning. Some angiographic studies of primitive hepatic neoplasms are presented. PMID- 6289193 TI - [Kinetic and clinical studies on a new cephalosporin: Cefotiam]. AB - Kinetic and clinical evaluation of cefotiam, a new cephalosporin, is reported. It was found that the drug is rapidly distributed to the tissues. Equilibrium between tissues and plasma is reached in about an hour. Some 90-91% of the dose administered is excreted in the urine, and accumulation does not occur. A clinical cure was obtained in 27 of a series of 35 patients (77.1%). Improvement was observed in 7 cases (20%). The antibiotic proved ineffective in the remaining cases (2.8%). Tolerance was excellent and there were no side-effects worthy of note. PMID- 6289194 TI - [Paget's disease of the vulva. Review and case report]. PMID- 6289196 TI - Simultaneous recording of somatic and dendritic field potentials and combined microiontophoresis in the rat Ammon's horn in situ: effects of GABA and acetylcholine. AB - In rats under urethane anesthesia, a twin set of multibarreled micropipettes was used to record simultaneously in situ the somatic population spike, the positive field and the dendritic negative field (EPSP) evoked by commissural stimulation, and to apply microiontophoretically GABAergic and cholinergic agents at each level. Relying on the typical somatic responses evoked by local application of GABA and acetylcholine (ACh), an electrode can be positioned in the pyramidal layer with good accuracy (in the range of 50 microns) and the pharmacology of the excitatory drive impinging upon the apical dendrite concomitantly investigated. PMID- 6289195 TI - [Value of the endocyte in detection of carcinoma of the endometrium]. PMID- 6289197 TI - Role of GABAergic mechanisms in the substantia nigra pars reticulata in modulating morphine-induced muscular rigidity in rats. AB - Systemic administration of morphine (15 mg/kg i.p.) induced a muscular rigidity in rats, which was recorded from the gastrocnemius-soleus muscle as tonic activity in the electromyogram (EMG). Administration of muscimol (12.5 or 25 ng) into the substantia nigra pars reticulata (SNR) antagonized the rigidity in a dose-dependent manner, whereas bicuculline enhanced it. It was concluded that GABAergic mechanisms in the SNR play an important role in mediating or modulating the expression of the morphine-induced rigidity, which is known to be due to an alteration of striatal function via opioid receptors located in this brain area. PMID- 6289198 TI - Choroid plexus, brain and kidney Na+,K+-ATPase: comparative activities in fetal, newborn and young adult rabbits. AB - We have studied maturation of brain barrier systems in fetal, newborn, juvenile and adult rabbits. We have compared choroid plexus and brain Na+,K+-ATPase levels in each age group, as well as serum and CSF Na+ concentration as a measure of the ability of the choroid plexus to generate a gradient from blood to CSF. The choroid plexus appears functionally mature at all ages studied, both in ability to produce a Na+ gradient and in ATPase levels. In contrast, brain ATPase levels rose markedly with age. Kidney ATPase measured for comparison showed a pattern similar to brain. PMID- 6289199 TI - The regulation of pyruvate dehydrogenase activity in rat hippocampal slices: effect of dichloroacetate. AB - The effects of dichloroacetate (DCA), an inhibitor of pyruvate dehydrogenase kinase, on the phosphorylation of the alpha-subunit of pyruvate dehydrogenase and on the activity of pyruvate dehydrogenase (pyruvate:lipoamide oxidoreductase (decarboxylating and acceptor-acetylating), EC 1.2.4.1, PDH) were investigated in rat hippocampal slices. Incubating hippocampal slices with increasing concentrations of DCA resulted in an increase in the active portion of PDH, without changes in the total PDH activity, as well as an increase in the in vitro phosphorylation of alpha-PDH. The effect of DCA on PDH activity was very rapid, being almost maximal after 5 min. These results indicate that DCA in the hippocampal slice preparation inhibits PDH kinase and consequently stimulates PDH activity by decreasing its endogenous state of phosphorylation. Moreover the time course of the effect of DCA suggests that the turnover rate of the phosphate group carried by alpha-PDH is very rapid and can be manipulated by altering PDH kinase activity. PMID- 6289200 TI - Caffeine down-regulates beta adrenoreceptors in rat forebrain. AB - We studied the influence of caffeine treatment (50 mg/kg for 3 doses) on catecholamine utilization and adrenergic receptor binding in female rats. Caffeine enhanced the reduction in forebrain norepinephrine levels following alpha-methyl-p-tyrosine without altering the reduction in dopamine levels. Caffeine reduced the apparent number of beta receptors in forebrain as measured by the Bmax for [3H]dihydroalprenolol binding. No changes in alpha1 or alpha2 receptor binding, as measured with [3H]prazosin and [3H]clonidine, respectively, were noted. These data show that caffeine selectively increases the rate of norepinephrine utilization in rat forebrain and that this is associated with a small, but significant, reduction in beta receptor density in this brain area. PMID- 6289201 TI - Receptor--ion channel interactions in Torpedo electric organ: influence of thiol group modification. AB - The influence of sulfhydryl and disulfide reagents on nicotinic cholinergic receptors and ion channels and their interactions was investigated using specific probes for receptor and channel binding sites in electric organs from Torpedo californica. N-ethylmaleimide, a sulfhydryl alkylating agent, did not alter receptor or ion channel binding, or receptor-mediated ion channel binding activation or desensitization. Alteration of receptor--ion channel coupling produced by treatment with an organic sulfhydryl, dithiothreitol, could be accounted for on the basis of decreases in receptor affinity for agonists. These changes were reversed by exposure to an oxidizing agent. Following reduction by dithiothreitol, N-ethylmaleimide treatment produced large decreases in the extent and affinity of both receptor and channel binding. No evidence for a direct role of thiol groups in ion channel function was obtained. PMID- 6289202 TI - Silicon overdosage in man. PMID- 6289204 TI - Wilms' tumor. PMID- 6289205 TI - Cefotaxime and moxalactam: the new third-generation cephalosporins. PMID- 6289203 TI - Studies of docosahexaenoic and eicosapentaenoic acids in trout and frogs. PMID- 6289206 TI - Neutralization of herpes simplex virus by antibody in amniotic fluid. AB - The ability of amniotic fluid to neutralize herpes simplex virus type 2 (HSV-2) was quantitated and compared with the serum neutralization titer in 158 pregnant women. Neutralizing activity was expressed as the ability of 0.1 ml of amniotic fluid to reduce the expected number of plaque-forming units (pfu) in the inoculum by 99%. All amniotic fluid samples from 32 women with serum titers of 1:40 or greater neutralized 5 pfu or more; at term, 96% of these fluid samples neutralized 50 pfu or more, 83% neutralized 500 pfu or more, and 61% neutralized 5000 pfu or more. Only 76% of the amniotic fluid samples obtained at term from women with serum titers of 1:5 to 1:39 contained detectable neutralizing activity and only 8% neutralized 5000 pfu or more. None of the amniotic fluid samples from 30 women with serum titers less than 1:5 neutralized 5 pfu or more. All neutralizing activity was removed when immunoglobulin G was removed from the amniotic fluid samples. Sera were obtained from 51 pregnant women 4 to 8 weeks prior to delivery. All women with serum titers of 1:40 or higher gave birth to infants who also had serum titers of 1:40 or higher. Therefore, it is possible to predict the neutralization titer in amniotic fluid and in the infant's serum by measuring the mother's titer in the third trimester. PMID- 6289207 TI - Gestational trophoblastic disease: treatment results at the Brewer Trophoblastic Disease Center. AB - Three hundred fifty-nine patients with gestational trophoblastic disease (choriocarcinoma and invasive mole) received complete treatment at the Brewer Trophoblastic Disease Center of Northwestern University Medical School from 1962 through 1978. Data were gathered as of December 31, 1978, to permit a minimum follow-up of 2 years. An overall remission rate of 92% was achieved: 100% (185/185) for nonmetastatic disease and 83% (144/174) for metastatic disease. All 200 patients with invasive mole and 129 of 159 patients (81%) with choriocarcinoma were cured. Chemotherapy was the main form of treatment, with adjuvant surgery and radiation therapy being used in selected patients. Five factors were determined to significantly influence response to treatment in patients with metastatic disease: 1) clinicopathologic diagnosis of choriocarcinoma versus invasive mole (71 versus 100%, P much less than .0005); 2) pretreatment human chorionic gonadotropin titer greater than 100,000 IU/liter and time greater than 4 months from pregnancy event to treatment (62 versus 93%, P much less than .0005); 3) metastases to sites other than lung and/or vagina (37 versus 92%, P much less than .0005); 4) antecedent term gestation compared with hydatidiform mole, abortion, and ectopic pregnancy (56 versus 79%, P less than .02); and 5) prior unsuccessful chemotherapy compared with no previous treatment (48 versus 83%, P much less than .0005). The value of secondary chemotherapy and adjuvant irradiation was evaluated. Relapse from remission was also studied. PMID- 6289208 TI - Clinical and virologic studies on female genital herpes. AB - Ninety women with genital herpes were studied clinically and virologically. From a clinical standpoint, these patients could be classified into 3 clinical types- acute, recurrent, and provoked. When herpes simplex virus (HSV) strains isolated from these patients and asymptomatic virus secretors were typed, 36 strains were determined to be type 1 (HSV-1) and the remaining 54 to be type 2 (HSV-2). The acute type was caused by primary infection of HSV-1 or HSV-2 as well as by infection of HSV-2 in the absence of heterologous antibody. In addition, 35 of 40 HSV strains isolated from recurrent, provoked, and asymptomatic virus secretors were identified as HSV-2, suggesting that HSV-2 may play a major role in establishing latent infection in the female genital region. PMID- 6289209 TI - [Treatment results in unselected small cell bronchial carcinoma patients]. AB - 18 of 36 consecutive patients with small cell carcinoma of the lung received a combination of chemotherapy (Adriblastin, Oncovin, Cyclophosphamide) and radiotherapy (primary tumor and CNS). This treatment resulted in a response rte of 78%, and in a median survival of 10.6 months ("limited disease" 13 months, "extensive disease" 9 months). Complete remissions were obtained only in 4 of 18 patients, 4 patients of these were long-term survivors (greater than 18 months). Due to heart diseases, age, or poor general condition 18 patients received a milder combination of chemotherapy ("COM", "VP-O-C") and tumor irradiation. The response rate was 66%. Complete remission was achieved in only 2 of 18 patients. The median survival rate was 8.6 ("limited disease" 10.5 months, "extensive disease" 6.0 months). Survival exceeded 18 months in 4 of these patients. The toxicity was mild, the mean cumulative duration of hospitalization was 78 (30 120) days per patient. PMID- 6289210 TI - Comparison of ultracytochemical methods for glucose-6-phosphatase activity in rat pigment epithelium. PMID- 6289211 TI - Ultracytochemistry in sections osmicated and embedded in epoxy resin and chemical analysis of chelated zinc on cytochrome c adjective reaction. PMID- 6289212 TI - Studies on intramitochondrial inclusions in the pancreatic acinar cells of the Japanese newt, Triturus pyrrogaster. II. The disappearance of intramitochondrial inclusions in rat-fed animals following starvation. PMID- 6289213 TI - Electron microscopic cytochemical analysis of paracrystalline intranuclear inclusions in human osteosarcoma cells. PMID- 6289214 TI - Effects of orchiectomy on the adrenal zona reticularis of adult rats: an ultrastructural morphometric study. PMID- 6289216 TI - Ultrastructural studies on filamentous inclusion bodies in the transitional epithelium of the monkey urinary bladder. PMID- 6289215 TI - Comparative-immunohistochemical study of the mammalian pituitary corticotrophs. PMID- 6289217 TI - Culture-proven cytomegalovirus retinitis in a homosexual man with the acquired immunodeficiency syndrome. AB - A 35-year-old homosexual man with cytomegalovirus viremia developed retinitis. He also had a new syndrome consisting of a persistent T-lymphocyte deficit, pneumocystis pneumonia, recurrent Candida albicans esophagitis, skin ulcerations caused by herpes simplex virus, Type 2, disseminated Mycobacterium avium intracellulare infection, and molluscum contagiosum. Histopathologic examination revealed bilateral necrotizing retinitis with virions in retinal, choroidal, and optic nerve tissues. Postmortem cultures of retina and vitreous were positive for cytomegalovirus. PMID- 6289218 TI - Cytomegalovirus retinitis in a young homosexual male with acquired immunodeficiency. AB - A case is reported of histopathologically documented CMV retinitis. It is part of a recently appreciated syndrome in young homosexual men, in which cellular immune deficiency has been documented and in which CMV infection may play a role. This case demonstrates that CMV retinitis is not excluded by negative CMV serology or cultures. PMID- 6289219 TI - Regression patterns of uveal melanomas after proton beam irradiation. AB - Forty-four uveal melanomas were evaluated for patterns of tumor regression after proton beam irradiation. All tumors were followed for a minimum of ten months after treatment. Seven lesions completely disappeared, 33 have decreased in size, and 4 remained unchanged. Associated signs of tumor regression were: resolution of the secondary serous retinal detachments, central apical yellow discoloration of the tumor, destruction of the tumor's vasculature, and elimination of fluorescein leakage. This study demonstrated that regression after proton beam irradiation is a relatively slow process, and the effects of radiation could be observed in some cases more than a year after treatment. PMID- 6289220 TI - Clinical and pathological studies in adult sheep and goats experimentally infected with Wesselsbron disease virus. AB - The clinical symptoms and pathology in 33 adult sheep and 31 adult goats experimentally infected with Wesselsbron disease virus are described. There was moderate to severe hyperthermia in most animals, but no other clinical signs of disease or deaths were recorded. Eleven sheep and 6 goats were sacrificed for pathological studies at various stages during the febrile response. The macroscopic and microscopic lesions in these cases are described. Microscopic studies revealed that the liver was consistently affected and showed small foci of necrosis. These were sparsely distributed and associated with a marked localized Kupffer cell response ("retothelial nodules"). In addition, acidophilic bodies and small groups of necrotic hepatocytes were evident in some lobules. Apart from the hepatic lesions, mild to moderate pyknosis and karyorrhexis of lymphocytes were seen in the spleen and lymph nodes. This report also compares the microscopic lesions in the livers of adult sheep and goats with those of new born lambs for Wesselsbron disease as well as with those reported for Rift Valley fever in adult sheep. PMID- 6289221 TI - Management of the primary dentition in vitamin D-resistant rickets. AB - Vitamin D-resistant rickets (familial hypophosphatemia) is a systemic disease secondary to defective renal-tubular reabsorption of phosphate. The major oral manifestations are spontaneous abscesses in a caries-free dentition. Pulpotomies or extractions were frequently described in the dental literature as the therapies of choice. This article presents a new prophylactic approach utilizing zinc oxide--eugenol pulpectomies and full crown coverage in an attempt to retain the primary dentition and prevent abscess formation. PMID- 6289222 TI - [Cytomegalovirus infection and hyaline membrane lung in children with acute leukemia]. PMID- 6289223 TI - [Possible causes of hemangiomatosis with reference to its clinical picture]. PMID- 6289224 TI - Aspirin clearly depresses responses of ventrobasal thalamus neurons to joint stimuli in arthritic rats. AB - This study dealt with the effect of aspirin upon activities of 17 ventrobasal thalamic neurons recorded in 17 rats rendered arthritic by injection of Freund's adjuvant into the tail. These neurons presented reproducible responses to mobilization and/or mild lateral pressure on a joint and were recorded for at least 40 min after aspirin administration. After intravenous injection of aspirin at the dose of 50 mg/kg (13 neurons tested), there was a progressive decrease in the number of spikes in the discharges. The maximum effect occurred at 30 min where the mean value of the response expressed as a percentage of the control was m = 34.62 +/- 7.5% (n = 13, p less than or equal to 0.001). Recovery was progressive and could be considered as complete at 60 min. By contrast, no significant modification of the spontaneous firing has been observed. With lower doses of aspirin (12.5 or 25 mg/kg tested with 4 neurons) there was respectively no clear depressive effect or only a transient decrease of the response. PMID- 6289225 TI - Macrophage requirement for host defence against experimental hepatic amoebiasis in the hamster. AB - The role of macrophages in hepatic amoebiasis in hamsters has been investigated by means of their treatment with bacille Calmette-Guerin (BCG) for activation, and with silica for elimination of these cells. Silica-treated animals inoculated intrahepatically with 1 x 10(5) trophozoites of Entamoeba histolytica developed amoebic abscesses in the liver and more metastases to other organs than control animals, and this effect was silica-dose-dependent. In contrast, BCG-treated animals developed significantly smaller abscesses in the liver and fewer metastatic foci. These data suggest that macrophages are involved in host defence against the establishment of amoebic liver abscess and metastatic dissemination of amoebae. PMID- 6289226 TI - [Penetration of cefotaxime into bronchial secretions]. AB - The objective of this study was to evaluate the penetration into bronchial secretions, of cefotaxime, a new highly-active cephalosporin. The study was performed in 45 patients with respiratory infections. The doses and the route of administration of the drug were different in 3 groups of patients: 10 patients received 0.750 g and 20 received 1 g intramuscularly; 10 patients received a 30 min IV Infusion of 2 g of cefotaxime. Bronchial samples were taken by means of fiberoptic bronchoscopy, after a single dose in all patients, and, respectively, after 3 and 7 days treatment in 30 and 15 patients. Simultaneous serum samples were collected in order to determine relationship between bronchial and corresponding serum levels. Assays were performed by means of the microbiological agar diffusion technique. In 30 cases bacteriological analysis was performed in order to determine the MICs for cefotaxime of the bacteria isolated in sputum. The results of the study showed a mean bronchial peak reaching about 2 microgram/ml at the 3d h. Individual concentrations were varying according to doses, route of administration and underlying pathology; the ratios between bronchial and corresponding serum levels were about 15 to 23 p. cent as usual for other cephalosporins. This study indicates that cefotaxime realizes significant bronchial amounts superior to MICs of microorganisms responsible for respiratory infections. PMID- 6289227 TI - [In vitro activity of ceftazidime compared with five beta-lactamase stable compounds against clinical strains of Acinetobacter calcoaceticus ]. AB - Acinetobacter calcoaceticus is recognized as one of the most resistant nosocomial pathogens. Clinical isolates of Acinetobacter are usually resistant to most beta lactam antibiotics. The objective of this study was to evaluate the in vitro activity of ceftazidime, a new broad spectrum highly potent beta-lactam antibiotic, able to inhibit especially Pseudomonas and Providencia. Its activity against 96 clinical strains of Acinetobacter was compared with the activity of 5 recent beta-lactam antibiotics which are resistant to beta-lactamase degradation (cefoxitine, Cefotaxime, moxalactam, cefotiam, cefamandole). The results of this comparative study of the in vitro activity of the 6 beta-lactam antibiotics exhibit a higher activity of ceftazidime: 50 p. cent of the strains were inhibited at a concentration of 4 micrograms/ml while the other drugs inhibited 50 p. cent of the strains at concentrations superior to 10 micrograms/ml; the geometric mean was 7 micrograms/ml for ceftazidime while for the other drugs it was more than 10 micrograms/ml and for 3 of them, the geometric mean was more than 40 micrograms/ml. Otherwise one could notice a bimodal distribution of the strains which suggests the presence of 2 populations of Acinetobacter, respectively inhibited with 4 micrograms/ml (susceptible) and 64 or 128 micrograms/ml (resistant). Finally this study shows that ceftazidime is one of the most active compounds against clinical isolates of Acinetobacter calcoaceticus among the 3rd generation of cephalosporins. PMID- 6289228 TI - [Ceftriaxone (Rocephine) in major African infectious pathology. Results at the Niamey Hospital (Niger) ]. AB - Ceftriaxone is a wide-spectrum-third generation cephalosporin characterized by outstandingly high efficacy as well as pharmokinetic properties making it suitable for administration in a single daily injection. Ceftriaxone has been found to be useful for treatment of the very severe infectious pathology in countries where hygiene and medical superstructures are still rudimentary. Eighteen of 20 patients with purulent meningitis (13 to Neisseria meningitidis A, 3 to Streptoc. pneumoniae, 1 to Listeria and 3 aseptic) recovered (there being 2 deaths at the 36th hour) after a mean 6 days of hospitalization. Despite the very delicate patient condition, recovery was seen in all 11 cases of very grave bronchopneumopathy, generally due to Streptoc, pneumonia. A dose of 2 g/day in 1 or 2 IV injections is sufficient in the adult, 0.50 g in a single dose being injected to infants weighing less than 10 kg, Meningitis required 4 to 7 days treatment (9 days in a case of Listeria) while the treatment period was longer for respiratory infections. Seven patients had been refractory to treatment with beta-lactamines and/or aminosides, and no adverse drug reactions were noted. PMID- 6289229 TI - [Influence of antibiotic disk charge on concordance curve of cefotaxime ]. AB - The activity of new cephalosporins is considerably higher than that of the first molecules. However antibiotic desk charge used for the agar diffusion method is still 30 micrograms. The disk diffusion - MIC distribution is not linear. The authors observed the influence of disk charge on the aspect of this distribution in order to determine the change permitting the best linearity. One hundred sixty five strains were studied using the agar diffusion and dilution methods. Their MICs were distributed from 0.015 to 128 mg/l. The disk charges studied were 5, 10, 15, 20, 25 and 30 micrograms. The results confirmed the parabolic aspect of concordance curve. This phenomenon was observed at higher disk charges (20, 25 and 30 micrograms). The linearity is better with the curves calculated from 5 and 10 micrograms. The correlation coefficients between these two charges were 0.92. However the inhibition diameters in 5 micrograms were too small to permit a clear differentiation between resistant and intermediate strains. The best discrimination is obtained with 10 micrograms. The zone diameter breakpoints were greater than or equal to 18 mm for susceptible 12 to 17 mm for intermediate, and less than or equal to 11 mm for resistant. PMID- 6289230 TI - [Bacteriological, pharmacological and clinical comparison between amoxycillin and ceftriaxone in the treatment of 300 purulent meningitis ]. AB - Two series of patients suffering purulent meningitis were treated: one (137 patients) with amoxycillin (200 mg/kg/day) by 4 intramuscular injections each day, the other with ceftriaxone (163 patients)/42 mg/Kg/day IM by intramuscular injections each day in the first 23 patients and then only one injection by day in the 140 other patients. Bacteriologically the superiority of CFTRX appears the whole studied strains: MIC of CFTRX are four times lower than those of AMOX for pneumococci, ten times lower for H. influenzae, hundred time lower for meningococci. Amongst the add strains the percentage of resistance to AMOX reaches 64 and only 7 to CFTRX. Pharmacologically, after a same dose of 50mg/kg the peak concentrations in CSF has the same level: CFTRX: 6.8 micrograms/ml, AMOX: 6 micrograms/ml. CSF levels remain efficient for 2 hours with AMOX and for 24 hours with CFTRX. The therapeutic index (mean antibiotic concentration in CSF/mean MIC of the strains) is higher with CFTRX than with AMOX (X 4 for pneumococci, X 15 for H. influenzae, X 100 for meningococci). Clinical results are better with CFTRX than with AMOX in each of the aetiological groups except meningococcal meningitis but the only significative difference concerns pneumococcal meningitis. Clinical tolerance of the two treatments was good. However 2 neonates treated with CFTRX has a severe eczematous erythrodermia and 8 other patients treated with CFTRX had diarrhoea due to elimination of the sensitive flora. PMID- 6289231 TI - Pathologic studies in colorectal cancer. A guide to the surgical pathology examination of colorectal specimens and review of features of prognostic significance. AB - The results of a careful and systematic examination of surgical specimens from patients with tumors of the colorectum play an important role in patient care and the assessment of prognosis. An accurate and detailed gross examination of the specimen is important. Photographs and line drawings are very useful. Proper handling of the specimen and fixation for 24 to 48 hours are essential before satisfactory blocks can be taken for sectioning. Lymph nodes adjacent to the lesion and at the point of ligation of the vascular pedicle should be removed for sectioning. Multiple blocks of the tumor and adjacent tissues should be taken for histologic study. All blocks should be appropriately labeled and properly identified. I have found the use of large sections of the entire lesion a very satisfactory method of studying cancers and polyps; in such preparations the anatomic relationships remain undisturbed. The pathology report should include information on the site or sites of tumor, the size, configuration, and circumference of the bowel wall involved, obstruction, distance of the resected margin from the tumor, depth of infiltration, tumor grade, tumor margin, local inflammatory reaction, lymph node involvement and location, and venous and perineural invasions. There is evidence in the literature to suggest that all of the above pathologic features are either essential or desirable for pathology reports and have varying degrees of prognostic value. The literature on the prognostic value of immunomorphologic features in nontumorous regional lymph nodes is contradictory. Cortical, paracortical, and sinus hyperplasia have been reported to be both associated with or to have no relation to prognosis. However, careful documentation of the changes in lymph nodes may in time shed light on their value, if any, with regard to survival. We have found a standardized pathology reporting system very useful in compiling data and evaluating patient prognosis. Moreover, this system of reporting can be used as a method of quality control in establishing minimum standards for data collection, as originally suggested by Buckwalter and Kent. In a regional multiinstitutional study such as ours, differences attributable to subjective variations among different observers are likely to occur, and may have some bearing when different pathologic features are eventually correlated with survival. PMID- 6289232 TI - The ultrastructure of malignant epithelial neoplasms of the lung. PMID- 6289233 TI - The pathology of tuberous sclerosis. PMID- 6289234 TI - Shedding and survival of herpes simplex virus from 'fever blisters'. AB - Shedding of herpesvirus by adults with herpes labialis and survival of the virus in the environment were examined. In nine adults with virus-positive herpes labialis, herpesvirus was detected in the anterior oral pool of seven (78%) and on the hands of six (67%). Herpesviruses isolated from patients with oral lesions were found to survive for as long as two hours on skin, three hours on cloth, and four hours on plastic. These findings support earlier recommendations for the protection of neonates from adults with "fever blisters." In addition, environmental surfaces may be a source of transmission of herpesvirus to the neonate. PMID- 6289235 TI - An outbreak of airborne nosocomial varicella. AB - An outbreak of nosocomial varicella was traced to airborne spread from an immunocompromised child hospitalized from Nov 11-19, 1980. Seventy potentially susceptible children were hospitalized on the ward during that period. Although the index patient remained in strict room isolation throughout his hospital stay, eight of these patients contracted varicella. The afternoon of November 12 was the period of highest risk for acquiring varicella. Eight of 36 patients (22%) present that afternoon, compared to none of 34 patients not present that afternoon, acquired the infection. A patient's risk of contracting varicella was significantly related to how near he/she came to the index patient's room that afternoon. Airflow studies, using the tracer gas, sulfur hexafluoride (SF6), demonstrated that patient rooms on this ward were at positive pressure with respect to the corridor. Despite isolation procedures, SF6 released in the index patient's room achieved concentrations in the corridor as high as 10% of those inside the room. Airborne spread of varicella has rarely been reported, but it may be a common mode of transmission in hospitals. We suggest that patients hospitalized with varicella be placed in strict isolation in negative-pressure rooms to reduce the risk of nosocomial transmission. PMID- 6289236 TI - Varicella zoster in hospital personnel: skin test reactivity to monitor susceptibility. AB - An outbreak of chickenpox, involving three patients and one nurse on a pediatric ward, necessitated rapid identification of susceptible employees in conjunction with standard epidemiologic intervention in order to prevent spread to other high risk patients. Of 46 hospital personnel 15 (33%) gave a negative or unknown history of prior disease. Response to a varicella-zoster skin test was compared with antibody determination as measured by fluorescent antibody to membrane antigen. Correlation of these two screening methods was absolute. Of 46 hospital personnel four (9%) were susceptible to infection (negative skin test and antibody less than 1:4) requiring their removal from the ward. All those with positive histories for prior disease with the virus and 11/15 (73%) with negative or unknown histories were immune as indicated by both tests. A readily available varicella zoster skin test would be an extremely useful epidemiologic tool for screening hospital personnel. PMID- 6289237 TI - [Neonatal hepatitis and growth hormone deficiency]. PMID- 6289238 TI - The magical number fourteen: making a very great deal of non-sense. PMID- 6289239 TI - The endocrine pancreas of the neonate and infant. PMID- 6289240 TI - Desensitization in denervated mouse muscles. AB - In denervated mouse soleus (DSOL) muscle preparations washed in Na methanesulfonate solutions containing 30 mmol X 1(-1) K+, bath-applied ACh (55 mumol X 1(-1) ) caused the resting potential to decrease from about -36 to -3.2 mV within 2-4 s; the potential remained stable or became slightly more negative when ACh was applied for 1- 2 min. Two components of the membrane current change (ACh current) were found in DSOL when using a point voltage clamp, an initial current component declining to 1/2 the initial amplitude in 11-15 s (desensitization) and a steady late current comprising 16-47% of the maximum ACh current. Membrane conductance (in microseconds X cm-2) was 0.35 in the absence of ACh, 20.3 at the peak of the initial current, and 1.57 during late current. The late current saturated at 0.55 to 5.5 mumol X 1(-1) ACh, whereas the initial current required 55 or more mumol X 1(-1) ACh to saturate. The null (reversal) potential was 6-13 mV more positive for the late current than for the initial current. The late current was masked when Na+ was replaced by Tris+, sucrose, or K+. An initial and a late current could also be distinguished in non-denervated endplates. The late current was more sensitive to ACh than the initial current, but the null potential was more negative than that for the initial current in endplates. In denervated membrane, the half time of desensitization was increased when Mg2+ was replaced by Ca2+ but the changes were on the average less than 15% the control values. It was concluded that desensitizing and non-desensitizing receptors may exist in extrajunctional membranes of denervated muscles and in endplates, the two being attached to different ionic channels. PMID- 6289241 TI - Temperature-dependence of 45Ca fluxes and contraction in vascular smooth muscle cells of rabbit ear artery. AB - The effect of cooling from 35 to 20 degrees C on the 45Ca-exchange and on the contractile response of rabbit ear artery has been investigated. The amplitude of the contraction induced by K-depolarization at 20 degrees C is reduced to about 60% of its value at 35 degrees C, whereas the response to noradrenaline is not significantly affected. Cooling induces a 2 to 4-fold reduction of the 45Ca efflux rate. This effect also occurs in Ca-free medium and in solutions containing 1 mM La. It also occurs in Na-free medium and in tissues in which the transmembrane Na-gradient has been reduced. At 20 degrees C, the 45Ca-influx in unstimulated tissues and in K-depolarized preparations is significantly lower than at 35 degrees C. In Ca-depleted tissues, i.e. tissues in which the noradrenaline-sensitive Ca-store has been emptied by a stimulation with the agonist in Ca-free solution, the 45Ca-influx is not significantly affected by cooling. The gradual depletion of the noradrenaline-sensitive Ca-store in Ca-free solutions is at 20 degrees C much slower than at 35 degrees C. The amount of Ca released by noradrenaline is not affected by cooling, whereas for the same amount of Ca released the contractile response is higher at 20 degrees C. These findings indicate that temperature affects the transmembrane Ca-extrusion and the Ca influx through voltage-dependent channels. The properties of the noradrenaline sensitive Ca-store are less sensitive to temperature. PMID- 6289243 TI - Prejunctional effects of anticholinesterase drugs at the endplate: mediated by presynaptic acetylcholine receptors or by postsynaptic potassium efflux? PMID- 6289242 TI - Cyclic AMP inhibits contractility of detergent treated glycerol extracted cardiac muscle. AB - Glycerinated myocardial fibres treated with a detergent (Lubrol WX) and suspended in ATP salt solution produce half maximum isometric tension at pCa 6.2 (at pH 6.7). After addition of cyclic AMP (1-100 microM), the pCa required for half maximum activation is 5.9. c-AMP in concentrations of 1-100 microM induces a dose dependent inhibition (up to 40% at pCa6), and this effect can be amplified by the phosphodiesterase inhibitor IBMX (3-isobutyl-1-methylxanthine) 10(-4) M. The effect is similar in presence and absence of sodium fluoride 10 mM. Since in detergent treated skinned fibres the cell membrane and the sarcoplasmic reticulum are extracted and since the Ca2+ ion concentration was kept constant and buffered, we propose that c-AMP does not act via the cell membrane or the sarcoplasmic reticulum, but via phosphorylation of troponin I. The latter is the only component which becomes phosphorylated in skinned fibres during c-AMP induced relaxation, an effect which is also responsible for the inhibition of actomyosin ATPase at constant Ca2+ ion concentration (cf. Ray and England 1976). PMID- 6289244 TI - Tizolemide-induced changes of passive transport components across the basolateral membrane of isolated frog skin. AB - The effect on transepithelial Na transport of tizolemide was investigated in isolated frog skin (Rana temporaria). It was found that tizolemide (2-5 mM, serosal side) decreased transepithelial Na transport (measured as short circuit current and as net sodium flux) within 60 min to 25-40% of the control level resulting from reduction of the unidirectional sodium influx. Intracellular recording with microelectrodes revealed that these changes were associated with depolarization of the intracellular space to less than 40% of the control values (averaging - 71.7 +/- 5.1 mV) which is a consequence of a decrease in conductance of the basolateral border to about 25% of the control values. The conductance of the apical border was only slightly reduced. It is suggested that tizolemide blocks the partial conductance of potassium at the basolateral border which secondarily diminishes transepithelial Na transport due to a decrease of the driving force for apical border Na entry. A certain degree of inhibition of the Na-K-ATPase by tizolemide cannot be excluded. When vasopressin (ADH) was added to frog skin after treatment with tizolemide, the response was markedly reduced compared to that of untreated control preparations. Under these conditions, the conductance of the basolateral border increased while the apical border remained little influenced by the hormone--opposite to the response of frog skins under control conditions. It is concluded that the mode of action of ADH is more complex than has been recognized hitherto and includes effects at the basolateral border. PMID- 6289246 TI - Single channel Ca2+ currents in Helix pomatia neurons. AB - Unitary Ca2+ currents of TEA injected Helix neurons were recorded in the "Giga seal" situation (6,7) from microscopic membrane patches exposed to 50 mM [Ca2+]o, O [Na+]o, 20 mM [TEA+]o and 2.5 microM [TTX]o. Constant field assumptions yield a channel permeability of 2.9 +/- 1.0 x 10(-14) cm3s-1 corresponding to slope conductances of 5 to 15 pS between 0 and -30 mV. Frequency of occurrence of the units strongly increased with depolarization. Mean open time of the Ca2+ channels was about 3 ms without obvious dependence on voltage. A similar open time was seen with [Ba2+]o, yielding about double the current strength when compared with [Ca2+]o. PMID- 6289245 TI - The time courses of intracellular free calcium and related electrical effects after injection of CaCl2 into neurons of the snail, Helix pomatia. AB - Controlled quantities of 100 mM aqueous CaCl2 solutions were pressure injected into voltage-clamped neurons with a resolution of 10(-11) 1. Ca2+-selective microelectrodes monitored the time course of changes in [Ca2+]i. At a membrane potential of -50 mV CaCl2 quantities in the range of 1% of the cell volume induced an inward current, associated with a conductance increase and having an equilibrium potential between -20 and +20 mV, which accompanied the rise in [Ca2+]i. An artifactual origin of the inward current by the injection procedure or by calcium screening of membrane sites could be excluded. The calcium-induced hyperpolarizing conductance, producing an outward current at -50 mV, followed the inward current and reached maximum during the late decline in [Ca2+]i. In most cases its development was separated from the inward current by an intermediate relative decrease of the membrane conductance. Neither of the two transient conductance increases showed a particular dependence on voltage. Renewed Ca2+ injection quickly decreased the calcium-induced hyperpolarizing conductance for several seconds. Ca2+ injections below 0.05% of the cell volume mostly produced pure outward currents or hyperpolarizing responses. Partial substitution of extracellular CaCl2 by NiCl2 decreased the hyperpolarizing response but not the initial inward current. The immediate effects of increased [Ca2+]i are activation of a depolarizing conductance and the partial block of the late hyperpolarizing conductance. The latter is probably produced through intermediate steps after increasing [Ca2+]i. PMID- 6289247 TI - Nigral output neurons are engaged in regulation of static fusimotor action onto flexors in cat. AB - Picrotoxin and muscimol were unilaterally injected into the postero-lateral part of the reticular zone of substantia nigra (plSNR) through chronically implanted guide cannulae in ketamine-anesthetized cats. Afferent activity of pretibial flexor muscle spindle primary and secondary endings was recorded before and after drug administration, and spindle sensitivity monitored during both sinusoidal and ramp stretch of the receptor-bearing muscle. From changes in spindle sensitivity after drug injection it is deduced that unilateral block by picrotoxin of the action of GABA on postsynaptic receptors in plSNR removes tonic static fusimotor action from flexor muscle spindle primary endings. Secondary endings seemed largely unaffected. The effect on primary endings is reversed by a subsequent injection of muscimol. It is concluded that the central nervous system, through GABA-modulated nigral output neurons, can control static fusimotor action onto flexor muscle spindle primary and secondary endings separately to some extent. PMID- 6289248 TI - Does the organic calcium channel blocker D600 act from inside or outside on the cardiac cell membrane? AB - The effects of extra- and intracellularly applied D600 (methoxyverapamil) and D890 (a quarternary derivative) on the action potentials of isolated guinea pig myocytes were compared. We also studied the extracellular myocytes were compared. We also studied the extracellular effects of these drugs on the calcium current (hybride sucrose gap) and contractile force of right ventricular trabeculae of the cat heart. The following results were obtained: 1. In ventricular trabeculae D600 suppressed the calcium current, tension and the plateau of the action potential. In contrast, D890 even in a 50 times higher concentration did not display any effect on these parameters. 2. In single isolated cells external application of D890 did not alter the configuration of the action potential. In contrast, external application of D600 suppressed the plateau and shortened the action potential in a dose-dependent way. 3. Intracellular injection of D600 or D890 strongly lowered the height of the plateau and abbreviated the action potential. The onset of the effects of both drugs was more rapid on intracellular application than that of external D600. Whereas the effect of an intracellular injection of D600 was reversible, that of D890 was not. These results support the hypothesis that the organic calcium channel blocker D600 enters the cell in the uncharged lipid soluble form and reaches its receptor associated with the calcium channel from inside. Because of its inability to pass the hydrophobic cell membrane, D890 is ineffective from outside but displays blocking effects on intracellular application. PMID- 6289249 TI - The effect of internal barium on the K current of the node of Ranvier. AB - Voltage clamp experiments were performed on single myelinated nerve fibres of the frog Rana esculenta to study the influence of internally applied Ba on the potassium outward current IK. Potassium currents of Ba-treated fibres decay rapidly during 30 ms depolarizations of more than 60 mV. The time constant tau of decay and the steady state potassium current IKss are strongly voltage dependent. The equilibrium dissociation constant K of the Ba-receptor complex decreases e fold for a 39 mV change of potential. The analysis suggests that the binding site is located at 30% of the membrane field seen from the inside of the axon. PMID- 6289250 TI - An analysis of the epileptogenic potency of CO2+- its ability to induce acute convulsive activity in the isolated frog spinal cord. AB - The action of Co2+ on the isolated frog spinal cord was studied by extracellular application of the ion in the superfusing solution. A complete and reversible blockade of chemical synaptic transmission by Co2+ (3 mmol/l) could be achieved after a superfusion period of 20-30 min. During continued Co2+ application (greater than 60 min) the following effects upon the motoneuron membrane, dorsal root and ventral root fibres were observed. Motoneurons and ventral root fibers: 1. prolongation of initial segment action potential to a maximum of 30 ms, 2. blockade of the long afterhyperpolarization, 3. abolition of adaptation, 4. increased duration of fibre action potential in the ventral root, 5. backfiring after ventral root stimulation. Dorsal root fibres: 1. prolongation of the extraspinal fibre action potential, 2. marked prolongation of the action potential of the terminal region, 3. backfiring of multiple action potentials after dorsal root stimulation. Even in the presence of Co2+, when synaptic transmission was completely blocked, strong convulsive reactions of the isolated spinal cord were observed. Intracellular injection of Co2+ into motoneurons did not affect the action potential, but led to a shift of the EIPSP towards the membrane potential. The results indicate that the induction of convulsive reactions by Co2+ is mainly due to a prolongation of action potentials. The plateau-like deformation of the action potential of the initial segment membrane and presumably of the terminal region of nerve endings results in retrograde propagation of action potentials and in some cases induces oscillatory discharge of single neurons. PMID- 6289251 TI - Voltage clamp analysis of the effect of cationic substitution on the conductance of end-plate channels. AB - The effect of two commonly used sodium substitutes, tris and glucosamine, on the amplitude and kinetics of miniature end-plate currents (MEPCs), acetylcholine (ACh) induced end-plate currents (EPC) and EPC fluctuations was studied in voltage clamped single muscle fibres from a monolayer preparation of the cutaneous pectoris muscle. Total replacement of sodium with each substitute shifted the reversal potential from -4.7 mV (normal sodium solution) to -3.6 mV (tris) and -49.0 mV (glucosamine). In tris and glucosamine substituted solutions the current (MEPC or EPC) - voltage relation became markedly nonlinear, with peak current decreasing with membrane hyperpolarization. Peak current at +40 mV, was unaltered in tris solutions and reduced in glucosamine substituted solutions. MEPCs decayed with a single exponential time course and the EPC fluctuation spectra were characterized by single Lorentzian functions in both normal sodium solution and each substituted solution. Analysis of EPC fluctuations demonstrated that both tris and glucosamine decrease single channel conductance and increase channel lifetime. Both effects were enhanced by either membrane hyperpolarization or by increasing the concentration of each substitute. In the presence of each cationic substitute, single channel conductance increased and mean channel lifetime decreased with membrane depolarization. Analysis of the data according to the constant field assumptions (Goldman, Hodgkin, Katz equation) provided an inadequate description of experimental currents obtained at hyperpolarized membrane potentials with total ion substitution. Shifts in reversal potential with partial substitution were, however, adequately predicted by the GHK equation. These results suggest that tris and glucosamine ions interact with end plate channels to reduce cation permeability and decrease channel closing rates. The dependence of this block on the level of membrane potential suggests that these cations bind to site(s) within open end-plate channels. PMID- 6289252 TI - Mechanism of adenosine 3',5'-monophosphate (cAMP)-induced increase in Ca2+ uptake by the sarcoplasmic reticulum in functionally skinned myocardial fibers. AB - The increased rate of Ca2+ uptake and ATPase activity in isolated cardiac sarcoplasmic reticulum (SR) by adenosine 3',5'-monophosphate (cAMP) has been shown to be activated by a cAMP-dependent protein kinase (cAMP kinase). Functionally skinned myocardial fiber preparations were used to study the mechanisms of cAMP action on the SR at the same time that tension was monitored. cAMP effects were studied on Ca2+ -activated tension of the contractile proteins, and on Ca2+ uptake and release from the SR using caffeine-induced tension transients. Neither cyclic AMP (0.1-5 microM) nor the catalytic subunit of cAMP kinase (0.1-1 microM) (PK-C) significantly changed either the maximal or the submaximal Ca2+ -activated tension. The areas of the tension transients were unchanged when cAMP was present in the releasing solution (release phase), and were significantly increased up to a mean of about 80% when cAMP or PK-C was present in the Ca2+ loading solutions (uptake phase). The increased tension transient was blocked by heat-stable inhibitor of cAMP kinase. We conclude that cAMP-induced increases in Ca2+ uptake by the SR could play an important role in the positive inotropic effect. cAMP kinase could thus play a crucial role in the regulation of myocardial contractility. PMID- 6289253 TI - Properties of a calcium- and voltage-activated potassium current in Helix pomatia neurons. AB - A calcium- and voltage-dependent current was found to be the principal outward current in identified Helix neurons. The current depends on the presence of [Ca2+]0, with half maximal activation at 1 mM [Ca2+]0, and it saturates beyond about 5 mM. The current is termed IK(Ca) since the charge carried by it corresponds to the amount of potassium ions transferred from the cell interior, as determined from the increase in K+ concentration in the external volume with K+ liquid ion-exchanger microelectrodes. IK(Ca) is characterized by cell shaped isochronal I/V curves. The peaks of these curves move from +30 mV to about +70 mV with an increase of the time of measurement from 30-200 ms. IK(Ca) rise times have a minimum of 10-15 ms at low depolarization around 0 mV, but increase about exponentially with more positive potentials. A tenfold decrease in [Ca2+]0 over the range of 30 to 0.3 mM also produces an increase in rise time, equivalent to a positive shift of potential by 20 mV. On repolarization of the membrane IK(Ca) disappears much faster than the intracellularly accumulated Ca2+, with a time constant which is similar to the minimum activation time constant. PMID- 6289254 TI - Activation characteristics of the calcium-dependent outward potassium current in Helix. AB - The activation of calcium-dependent outward potassium current [IK(Ca)] by shortlasting Ca2+ inward currents was studied. These Ca2+ currents were produced either by small depolarizing pulses preceding the larger depolarizations or by interposed repolarizations (I.R.s) starting from depolarized membrane potentials. IK(Ca) then develops with a potential-invariant time course (half time 6-12 ms) and the normally bell-shaped isochronal IK(Ca)/V curve, measured at between 30 and 300 ms, is straightened. However, there Ca2+ - injecting pulses, of any amplitude and duration, do not increase the steady-state conductance to values beyond those measured with single step depolarizations to lower potentials. varied in length, activation of IK(Ca) increased linearly with the during depolarization to near the supposed calcium equilibrium potential with no further Ca2+ influx. When I.R.s are varied in length, activation of I(Ca) increases linearly with the amount of Ca2+ current. Fading of activation during I.R. follows a time course nearly ten times slower than activation and is not expressed in tail currents. The time course of IK(Ca) is described by a function defined only by voltage parameters of activation combined with a minimum activation time constant which is similar to that found in tail currents. Peak location and general form of the IK(Ca)/V relationship for times up to several hundred milliseconds are well predicted without the necessity of explicitly account for actual calcium entry. PMID- 6289255 TI - Effects of barium on the membrane currents in the rabbit S-A node. AB - In small preparations of rabbit sinoatrial node voltage clamp experiments with the two microelectrode technique were carried out. The effects of extracellular barium ions on the slow inward current and outward currents were studied and the following results were obtained: 1. Ba increased the amplitude of the slow inward current without a change in the time course of inactivation. In 10 mM Ba the steady-state inactivation curve (f infinity) shifted in the positive direction (3 4 mV), suggesting a neutralization of negative surface charges. A similar shift of the steady state activation curve (d infinity) could not be detected. 2. Ba reduced the amplitude of the time-dependent (IK, Ix) and time-independent (Ibg) potassium currents in a concentration-dependent manner. 3. The block of the time dependent potassium current, IK, depended on the membrane potential. The block was stronger at negative than at positive potentials. The block could be relieved by depolarizing pulses, the degree of unblock increased with longer duration of the depolarizing pulse. 4. Ba blocked the slow outward current Ix in a voltage- and time-dependent manner. Like for IK, the block of Ix was stronger at negative than at positive potentials. A given concentration of Ba produced stronger block of Ix than of IK and the removal of block of Ix by depolarizing pulses was slower than the removal of IK block. 5. The effects of Ba on Ix suggest that this current is a potassium current. PMID- 6289256 TI - Conductance of the slow inward channel in the rabbit sinoatrial node. AB - The elementary conductance of the slow inward current channel in the rabbit sinoatrial node was measured by analysis of the current fluctuations. The preparations were voltage-clamped to -30 mV, where d infinity and f infinity of the slow inward current (Isi) intersect. In the presence of Ba, which increases Isi and decreases outward currents, a small steady-state component of Isi could be detected. The fluctuations of the current in 10 mM Ba were smaller due to the block of the outward channels. They were further reduced after the inhibition of Isi by the Ca channel blocker D600. The spectral power density distribution of the current fluctuations originating from Isi could be fitted at frequencies less than 30 Hz with a single Lorentzian which was attributed to the inactivation process. The corner frequency was 5.28 +/- 1.16 Hz (n = 10), corresponding to an average open time of the single channel of about 30 msec at -30 mV. The single channel conductance was determined to 6.50 +/- 3.15 pS (S.D., n = 10). PMID- 6289257 TI - External K+ ions increase rate of opening of outward current channels in snail neurons. PMID- 6289258 TI - Effect of low potassium-diet on Na-K-ATPase in rat nephron segments. AB - Na-K-ATPase activity was determined in 10 segments of the rat nephron using a fluorometric microassay method [4]. The enzyme activity showed three peaks (greater than 200 pmol ADP min-1 mm-1) along the nephron of normal rats. These peaks were in the S1 portion of the proximal tubule, the medullary thick ascending limb from the inner stripe and the distal convoluted tubule. Feeding the rats a low potassium diet for 8 weeks produced a significant decrease in Na-K ATPase activity in the cortical collecting duct, but no significant change in this enzyme in any other segment. The low potassium diet did not produce a significant change in Mg-ATPase in any nephron segments. We conclude that Na-K ATPase activity along the rat nephron shows a pattern that is qualitatively similar to that seen in the rabbit nephron [4]. However, quantitatively the Na-K ATPase activity in the rat nephron is greater than in the corresponding segments of the rabbit nephron. The results are consistent with the greater rate of glomerular filtration and Na+ reabsorption per rat nephron. Furthermore, our results suggest that the decrease in potassium excretion during potassium deficiency is modulated, at least in part, by the level of Na-K-ATPase activity in the cortical collecting duct. PMID- 6289259 TI - Patch-voltage-clamp method for measuring fast inward current in single rat heart muscle cells. AB - A patch-voltage-clamp method was used to measure fast inward ionic currents in single heart muscle cells. 1. Theoretical analysis including computer simulation has shown that the method provides fast settling of membrane potential (within 10 microseconds) and reliable voltage clamp on the tested membrane patch when the area of the patch is 200-300 times smaller than the area of the whole cell membrane. 2. The transient time at the I-V converter output is increased up to 70 75 microseconds due to the stray capacity in the I-V converter feedback. When fast-response operational amplifiers are used in the set up this transient time may be decreased to 30 microseconds by partial restoration of the high-frequency components of the signal. 3. Experimental data have shown that the ionic channel population in the membrane patch of about 5 micrometers in diameter is on the one hand large enough to directly observe integral ionic current and on the other hand small enough for the fluctuations of ionic current to be appreciable. This permits the method to be applied to ionic current investigations both by classical methods and by statistical analysis. PMID- 6289260 TI - Calcium channel block and recovery from block in mammalian ventricular muscle treated with organic channel inhibitors. PMID- 6289261 TI - [Computed tomographic and ultrasonographic diagnosis of portal vein tumor thrombus in hepatocellular carcinoma]. PMID- 6289262 TI - [Biological studies of proton irradiation under combination with a hypoxic cell sensitizer]. PMID- 6289263 TI - Processing of eukaryotic tRNA precursors: secondary structure of the precursor specific sequences affects the rate but not the accuracy of processing reactions. AB - The primary transcriptional product of eukaryotic tRNA genes is a precursor molecule with extranucleotides at the 5' and at the 3' end. We show that the 5' and 3' sequences, uniquely present in the RNA precursor molecule do not play any role in the efficiency and accuracy of processing reactions. If, however, as a consequence of in vitro manipulation, these extranucleotides form a base-paired extension of the aminoacid acceptor stem, the rate of processing is slowed down. The rate of processing is brought back to normal in a single base-pair deletion mutant probably as a consequence of a destabilization of the base-paired extension of the aminoacid acceptor stem. PMID- 6289264 TI - On the operon structure of the nitrogenase genes of Rhizobium leguminosarum and Azotobacter vinelandii. AB - The transcription of the nitrogenase genes in Rhizobium leguminosarum was studied by analysing total cellular RNA from bacteroids for the presence of nitrogenase messenger RNA. The RNA was separated by agarose gel electrophoresis and blotted onto nitrocellulose filters. Messenger RNA for nitrogenase was detected by hybridization with probes derived from plasmid pSA30, a recombinant plasmid carrying the nitrogenase genes of Klebsiella pneumoniae. In the same way nitrogenase mRNA was detected in RNA isolated from Azotobacter vinelandii and from Klebsiella pneumoniae, but only if both were cultured under conditions of derepression of the nitrogenase genes. The size of the RNA hybridizing with the probes indicates that in all three organisms the genes for the subunits of the two components of nitrogenase constitute a single operon. PMID- 6289265 TI - Glutamate tRNA genes are adjacent to 5S RNA genes in Drosophila and reveal a conserved upstream sequence (the ACT-TA box). AB - In Drosophila melanogaster at least six transfer RNA genes are located adjacent to the 3' end of the 5S RNA gene cluster. Three of these have been sequenced and identified as coding for glutamate tRNA4. In the chromosome they are arranged as tandem repeats on the same DNA strand and transcribed in the same direction as is 5S DNA, towards the centromere. We have also identified a sequence, the ACT-TA box, that is highly conserved among the polymerase III transcribed genes. Usually the sequence is located at 37 +/- 8 base pairs upstream from the first nucleotide of the structural gene. A similar sequence is also observed upstream of yeast and silkworm tRNA genes and the mitochondrial tRNA genes of mouse and humans. PMID- 6289267 TI - Ribosomal RNA genes in Mycoplasma. AB - Using Southern blotting analysis with labelled mycoplasmal ribosomal RNA as probe, two fragments (1 Kb and 5 Kb) were detected in an EcoR I digest of Mycoplasma capricolum DNA. This analysis revealed that the 5 Kb fragment carries both 16S rRNA sequences and the entire 23S rRNA gene of this mycoplasma. The 1 Kb fragment contains 16S rRNA sequences only. The 5 Kb EcoR I fragment has been cloned and used to characterize the structure of rRNA cistrons in various Mycoplasma strains. These experiments clearly demonstrate a substantial homology of Mycoplasma capricolum rRNA sequences with the E. coli rRNA cistron on one hand, and with Mycoplasma mycoides subsp. capri and Acholeplasma laidlawii on the other hand. This analysis also reveals two rRNA cistrons in Mycoplasma mycoides subsp. capri and Acholeplasma laidlawii whereas one rRNA cistron is present in Mycoplasma capricolum. PMID- 6289268 TI - Determination of the transcription initiation site of Tetrahymena pyriformis rDNA using in vitro capping of 35S pre-rRNA. AB - Approximately 700 nucleotide sequences surrounding the transcription initiation site were determined with a cloned rDNA fragment of Tetrahymena pyriformis and the transcription initiation site was localized on these sequences using purified 35S pre-rRNA. A considerable portion of the 35S pre-rRNA was found to be capped in vitro. The 32P-labeled, capped 35S pre-rRNA, on nucleus P1 protection mapping, gave the protection band which is identical in size with that obtained with bulk 35S pre-rRNA. Both reverse transcription extension and nuclease P1 mapping localized the 5'-end of the 35S pre-rRNA at the same adenine nucleotide, 496 base pairs upstream from the HindIII site of the cloned rDNA fragment. Furthermore, sequencing of the 5'-terminal region of the in vitro capped 35S pre-rRNA unambiguously confirmed the above result. The strategy adopted in the present experiment could serve as a general procedure for determining the transcription initiation point even in cases where the concentration of the primary transcript is low. PMID- 6289266 TI - Immunoglobulin switch region-like sequences in Drosophila melanogaster. AB - We found immunoglobulin switch (S) region-like sequences in DNAs of wide variety of organisms including sea urchin, yeast and Drosophila that do not produce immunoglobulins. DNA fragments carrying Smu-like sequences were cloned from Drosophila and the nucleotide sequence of a clone is almost identical to that of the mouse Smu region. Restriction fragments of Drosophila Smu-like sequences and their flanking regions seem to vary among Drosophila species. Possible evolutionary significance of the Smu-like sequence in invertebrates was discussed. PMID- 6289269 TI - Restriction fragment length polymorphism of the rat albumin gene in Sprague Dawley rats and its application in genetic study of analbuminemia. AB - Restriction fragment length polymorphism of the rat albumin gene was discovered in a stock of Sprague-Dawley rats by Southern blots of rat liver DNAs using cloned albumin cDNA, prAlb-1 (1), as a probe. The polymorphic DNA fragments were observed when rat DNAs were digested with either Hind III or Pst 1 and the difference in length of the DNA fragments in Hind III or Pst 1 digests was estimated as 1.4 kbp. When DNAs were digested with EcoR I, restriction fragment length polymorphism was not observed. Therefore, this polymorphic DNA was concluded to be located in the flanking sequence. Structural analysis of the cloned albumin gene showed that the polymorphism was located in the 3'-flanking sequence. With this polymorphism as a marker of the albumin structural gene, the phenotype of analbuminemia, which is an autosomally recessive trait, was found to be linked to the structural gene of albumin. PMID- 6289270 TI - Rearrangement of repeated DNA sequences during development of macronucleus in Tetrahymena thermophila. AB - Three clones of non-repetitive sequences and six clones containing repetitive sequences were obtained from micronuclear DNA of Tetrahymena thermophila. All the non-repetitive and three repetitive sequences had the same organization in micro- and macronuclear DNAs as revealed by blot hybridization. On the other hand, the remaining three clones with repetitive sequences had apparently different organization in the two nuclear DNAs. All these repetitive sequences showed a smear on the blot in addition to a number of discrete bands when micronuclear DNA was digested with EcoR I. In macronuclear DNAs, these sequences invariably became one or two bands and the smear disappeared. We conclude that, when a macronucleus develops from a micronucleus, the non-repetitive sequences amplify by more than 20 times with relatively few rearrangement, whereas some selected portions of repeated and/or repeat-contiguous sequences are amplified with rather extensive reorganization. PMID- 6289271 TI - Different levels of DNA modification at 5'CCGG in murine erythroleukemia cells and the tissues of normal mouse spleen. AB - The DNA of a Friend erythroleukemic cell line (clone DS-19) and that of mouse spleen from which it was derived were compared with respect to modification at 5'CCGG. Methylation patterns in clone DS-19 DNA were very different from those in normal spleen DNA. Our results suggest that the mechanism by which the internal cytosine in the 5'CCGG sequence is modified has been partially inactivated in clone DS-19. We observed a general decrease of about two-fold in total modification at this site in DS-19 DNA. This general decrease was shown to extend to 5'CCGG sites in specific classes of repeated sequences with two-dimensional displays of restriction fragments. Interspersed repeated sequences which are concertedly modified (i.e., modified at many different chromosomal locations) in spleen, were found to be unmodified at these locations in the cell line. However, we did not detect a difference in DNA modification at 5'CCGG with these techniques when uninduced and hexamethylene-bis-acetamide (HMBA) induced cells of clone DS-19 were compared. In other experiments, we obtained further evidence that selected classes of repeated sequences in Physarum polycephalum are methylated to the same extent independent of chromosomal location. PMID- 6289272 TI - Characterization of a rat tRNA gene cluster containing the genes for tRNAAsp, tRNAGly and tRNAGlu, and pseudogenes. AB - The putative genes for tRNAGAUAsp(C), tRNAGGAGly(G) and tRNAGAGGlu are in a cluster on the rat chromosome and are present exclusively in a 3.3 kb region cleaved with a restriction endonuclease EcoRI. The cluster reiterates about 10 times on the haploid DNA. Four lambda clones each containing an independent repeating unit were isolated from a rat gene library. The studies on the cloned DNA revealed that the length of the repeating unit including the 3.3 kb EcoRI fragment was at least 13.5 kb. Nucleotide sequence analysis of the 3.3 kb DNA in the isolated clones showed sequence variations among the repeating units and incomplete genes for tRNAGly and tRNAGlu within the clusters. PMID- 6289273 TI - [Advances in the physiopathology, diagnosis and therapy of bronchial asthma]. PMID- 6289274 TI - Myocardial damage following transthoracic direct current countershock in newborn piglets. AB - The effect of transthoracic direct current countershock on the myocardium of 21 newborn piglets was studied. Myocardial damage was quantified by measuring the myocardial uptake of technetium-99m pyrophosphate injected 24 hours after countershock. Substantial myocardial damage occurred in animals given greater than 150 joules/kg but not at lower energy doses. Damage occurred in both ventricular free walls, but more frequently in the right ventricle. The epicardial half of the myocardium was more severely affected than the endocardial half. The relationship between myocardial damage and total countershock energy dose was well modeled by an exponential function. Transthoracic direct current countershock appears unlikely to cause myocardial damage in newborn piglets unless greatly elevated energy doses are employed. PMID- 6289275 TI - Bioassays in modern peptide research. AB - Bioassays have been used in pharmacology since its beginning, nearly one hundred years ago. With the recent development of new techniques such as radioreceptor assays, radioimmunoassays, high pressure liquid chromatography (HPLC) and recombinant DNA technology, it might be asked if bioassays are still up-to-date in a fast moving field like peptide research? This brief review describes the possible uses of bioassays as well as the basic criteria used to characterize a bioassay. Recent results obtained with bioassays for various peptides will demonstrate the importance of this technique in modern peptide research. PMID- 6289276 TI - Correlated cholinomimetic-stimulated beta-endorphin and prolactin release in humans. AB - Previous studies, both in animals and humans, have demonstrated that the intravenous or intraventricular administration of endogenous opioids and opiates produce dose dependent increases in plasma concentrations of prolactin. Notably, in humans, intravenous infusion of centrally active cholinomimetic drugs, such as physostigmine or arecoline, may produce significant increases in plasma concentrations of prolactin and beta-endorphin immunoreactivity. In three separate studies, conducted collaboratively between the National Institute of Mental Health and the University of California at San Diego, physostigmine and arecoline associated increases in plasma concentrations of beta-endorphin immunoreactivity were highly correlated with increases in plasma prolactin concentrations. These results are of interest because centrally active cholinomimetic drugs have been variously reported either to have no effect, to increase, or to inhibit anterior pituitary prolactin release. We propose that cholinergic stimulation of hypothalamic beta-endorphin may represent an interesting example of peptidergic modulation of primary neurochemical effects on hypothalamic-pituitary hormonal regulation. PMID- 6289277 TI - The opiomelanotropinergic neuronal and endocrine systems. PMID- 6289279 TI - Evidence for an opiomelanotropin acetyltransferase in the rat pituitary neurointermediate lobe. AB - Results of this study demonstrate two distinct forms of acetyltransferase activity which will acetylate alpha-MSH. These acetyltransferases are distinguished by pH optima, subcellular distribution and sensitivity to magnesium and several solubilizing detergents. A general acetyltransferase, as characterized in the rat pituitary anterior lobe and lens, has a pH optima of 7.4 and is inhibited by magnesium. Subcellular fractionation of anterior and neurointermediate lobes revealed that this acetyltransferase is primarily localized in the cytosol fraction of these tissues. An alpha-MSH acetyltransferase (MAT) and a beta-endorphin acetyltransferase (EAT) have a pH optima of 6.0-6.6, are inhibited by detergents, and are specifically localized in the secretory granules of the neurointermediate lobe. Comparative studies of MAT and EAT suggest a single enzyme responsible for the acetylation of opioid and melanotropin peptides, and we term this enzyme opiomelanotropin acetyltransferase (OMAT). PMID- 6289278 TI - Pro-opiocortin processing in the pituitary: a model for neuropeptide biosynthesis. AB - The biosynthesis of alpha-MSH, beta-endorphin and ACTH in the pituitary is reviewed. These neuropeptides are synthesized from a common pro-protein, pro opiomelanocortin. The pro-protein is cleaved intragranularly, at pairs of basic residues in the molecule to yield the respective peptide products. An unique, thiol protease (pro-opiocortin converting enzyme) and a carboxypeptidase B-like enzyme, both localized within pituitary secretory granules and having a pH optimum of 5-6, appear to be involved in the proteolytic processing of pro opiomelanocortin. PMID- 6289280 TI - Central nervous system and peripheral effects of ACTH, MSH, and related neuropeptides. AB - Adrenocorticotropic Hormone (ACTH), Melanocyte-Stimulating Hormone (MSH), and related peptides have been shown to have several neurogenic effects: alteration of cerebral protein synthesis, RNA synthesis, protein phosphorylation, and neurotransmitter turnover. Furthermore, there appears to be an ACTH containing circuit in the CNS which originates in the arcuate nucleus. Changes in concentration of the peptides in this family have been shown to alter electrophysiology, neuromuscular function, and behavior (e.g., grooming, learning) in infrahuman subjects. These findings suggest that the neuropeptides MSH and ACTH influence the capacity of an organism to efficiently evaluate information and influence the affective functioning of humans. PMID- 6289281 TI - Endorphinergic and alpha-noradrenergic systems in the paraventricular nucleus: effects on eating behavior. AB - Brain cannulated rats were injected with the opioid peptide beta-endorphin (beta EP) directly into the hypothalamic paraventricular nucleus (PVN) where norepinephrine (NE) is most effective in stimulating eating behavior. Beta Endorphin (1.0 nmole) reliably increased food intake in satiated animals, and this response was blocked by local administration of the selective opiate antagonist naloxone. The eating induced by beta-EP was positively correlated in magnitude with the NE response and, like NE, was antagonized by PVN injection of the alpha-noradrenergic blocker phentolamine. Naloxone had no effect on NE induced eating, and the dopaminergic blocker fluphenazine failed to alter either beta-EP or NE eating. When injected simultaneously, at maximally effective doses, beta-EP and NE produced an eating response which was significantly larger than either of the responses elicited separately by beta-EP or NE and was essentially equal to the sum of these two responses. The evidence obtained in this study suggests that beta-EP and NE stimulate food ingestion through their action on PVN opiate and alpha-noradrenergic receptors, respectively, and that beta-EP's action is closely related to, and in part may be dependent upon, the PVN alpha noradrenergic system for feeding control. PMID- 6289282 TI - Peptide influences on the development and regeneration of motor performance. AB - ACTH 1-39 (0.2 U IP daily for up to 18 days) has a beneficial effect on the functional reorganization of regenerating motor units of the extensor digitorum longus (EDL) in the adrenalectomized adult rat following crushing of the peroneal nerve. Motor unit activity (maximum twitch tension amplitude/mean increment in twitch tension as voltage is increased by 0.1 V gradations) and nerve-muscle efficiency (tetanic tension from indirect stimulation/tetanic tension from direct stimulation of EDL) were enhanced by ACTH 1-39. Other electrophysiological and contractile parameters were unaffected by the peptide. Spontaneous motor activity in cold stressed 13 day old rats was prolonged by Org 2766, a substituted analogue of ACTH/MSH 4-9, (0.1 microgram/kg daily) but unaffected by the same dosage of ACTH/MSH 4-10. The responsiveness of developing and regenerating motor systems to neuropeptides indicates a plasticity of neuronal connections, which depends on peptide sequence, dosage and the physiological state of the animal (normal, depressed, regenerating or developing, at rest or stressed). PMID- 6289283 TI - Proteolysis of beta-endorphin in brain tissue. AB - The processing of beta-endorphin by brain enzymes into peptides related to the behaviorally active gamma- and alpha-type endorphins and the sequence of proteolytic events in the conversion process are described. Multiple enzyme activities contribute to the generation of the peptides with neurotropic activity. It is proposed that the processing into gamma- and alpha-type neuropeptides is a post-secretional event. The enzymes involved may have a key role in the nature and levels of neurotropic beta-endorphin fragments in the brain. PMID- 6289284 TI - Effects of desmopressin acetate (DDAVP) on the learning of a brightness discrimination. AB - Sixty male albino rats received DDAVP, a placebo, or control treatment and were tested on a brightness discrimination task. Three groups (DDAVP, placebo, and control) were tested in the morning and three groups were tested in the evening. The acquisition and reversal of the brightness discrimination, along with the retention of the reversal problem after a 5-day retention interval were analyzed. Inspection of forward and backward learning curves plotted for each task revealed facilitated acquisition along with an initial impairment of reversal learning in those animals treated with DDAVP. These results support a memorial interpretation of DDAVP's effects. This was short-term in duration, as no retention effects were obtained. It was also found that DDAVP's effects were not influenced by diurnal processes. PMID- 6289285 TI - The role of central and peripheral cholecystokinin in mediating appetitive behaviors. AB - Cholecystokinin (CCK) reduces total food consumption in mice, rats, pigs, sheep, monkeys and humans. Behaviors associated with an underlying state of satiety are reported after CCK administration. Reductions in exploration and social interactions by CCK are not due to true sedation or sleep, as measured by cortical EEG recordings. The satiety effects appear to be mediated by peripheral CCK receptors, through a feedback loop involving the vagus nerve. The conceptual link between the behavioral functions of CCK as a putative satiety signal and its established digestive functions are discussed. PMID- 6289287 TI - [Electrophysiological studies of peripheral nerves in children with various grades of chronic renal failure]. PMID- 6289286 TI - Modulation of [3H]-dopamine binding by cholecystokinin octapeptide (CCK-8). AB - Cholecystokinin-octapeptide (CCK-8) is a putative neurotransmitter which has been demonstrated previously to occur in midbrain dopamine neurones. We observe that CCK-8 causes changes in both the affinity and density of binding sites for [3H] dopamine in rat striatal homogenates, in vitro, upon incubation with the peptide at a concentration of 1 micromolar. A dose-response study of the competetion of CCK-8 with [3H]-dopamine binding indicates an IC50 for the peptide of 450 nM; desulfated CCK-8 and the related peptide caerulin are at least 4-fold less active than CCK-8. CCK-8 was also administered to rats in a separate study; the binding of [3H]-dopamine was evaluated to homogenates of striata and olfactory tubercles obtained from these animals, which had been treated with systemic injection at a dose of 20 micrograms/kg, daily, for four days. A decrease in the number of striatal binding sites for the radioligand was observed, with a concomitant increase in the number of binding sites in the olfactory tubercle. These data collectively suggest a possible regulatory role for CCK-8 in the ascending dopamine systems. PMID- 6289288 TI - Histological classification of gastric cancer from initial to advanced stages. AB - An ideal classification of malignant tumors should imply not only the histogenesis of the tumor and adequate reproducibility of its specific characteristics by other investigators, but also an exact definition of the stages of tumor development to facilitate prognostic prediction. The present paper proposes a classification of gastric carcinoma with emphasis on the initial stages of malignant transformation. The classification is based on the analysis of early invasive gastric cancer observed in biopsy material from the Institute of Pathology of Munster University. Since 1974, all cases of severe dysplasia or early gastric cancer were selected from a total of 200,000 gastric biopsies and more than 1500 gastric resections, and subjected to special work-up. In analogy to Clark's classification of malignant melanoma (Clark et al., 1969) the proposed classification defines consecutive stages of development in gastric carcinoma with special regard on the maximum depth of invasion and vertical thickness of the tumor, together with its histologic type and the predominant direction of tumor spread. PMID- 6289289 TI - Ultrastructure of peripheral lymphocytes in generalized ceroid-lipofuscinosis. Report of a case. AB - Electron microscopic observation was made on the peripheral blood cells obtained from a 4-year-old boy with generalized ceroid-lipofuscinosis (GCL), and two kinds of significant inclusions were found in the cytoplasm of lymphocytes. One was ceroid-lipofuscin accompanied with the fingerprint pattern, and the other was a lamellar body which showed the concentric lamellar structure or the myelin figure. Ceroid-lipofuscin was found in 40% of peripheral lymphocytes. It is important in diagnosing GCL that ceroid-lipofuscin can be detected with high incidence in lymphocytes of peripheral blood. PMID- 6289290 TI - CEA positivity in sera and breast tumor tissues obtained from the same patients. AB - It is well known that the concentration of serum CEA correlates best with colorectal carcinomas but less well with other solid tumors. The aim of the present study was to elucidate further the poor relationship between confirmed primary mammary carcinomas and CEA serum levels, 108 of these tumors have hence been studied for their tissular CEA positivity and the results obtained were compared with the CEA serum values found preoperatively in the same patients. Techniques employed were the indirect immunoperoxydase method for the histologic studies and the enzymoimmuno-assay for the serum measurements. The discordance and concordance of the CEA results obtained with the two techniques were analyzed in respect to the histologic types of tumors classified following the ICD-O nomenclature. Our results seem to prove that: 1) although most tumors produce the antigen only a fraction of them releases it, and 2) the most common histologic type of tumor is the one secreting the antigen the less frequently. These observations are apparently independant of the tumor stages T1-T4, N- or N+, M-. PMID- 6289291 TI - Chronic constipation in children: can it be managed by diet alone? AB - Chronic constipation is a common childhood problem. In a study of 60 children aged 2 to 12 years, a special diet that involved daily intake of raw bran and high-fiber foods and exclusion of milk and other constipating foods was found to be successful. Within six weeks, the problem resolved in all 60 patients. Milk and other restricted foods were returned slowly to the diet if they were found not to be a contributing factor to constipation. We believe our clinical observations support the prescription of a special diet for childhood constipation. For patients in whom specified organic causes have been excluded, we recommend a trial of this diet before other, more complex therapeutic efforts are undertaken. Implementation of the diet necessitates careful explanation to the child and family, as well as long-term follow-up. PMID- 6289292 TI - Simple purification and some properties of beef spleen exonuclease. AB - A method for purification of beef spleen exonuclease is described, leading to electrophoretically homogeneous enzyme preparation. The method consists of three step fractionation of crude enzyme (after ammonium sulfate precipitation) as follows - ion exchange chromatography on ECTEOLA-cellulose, affinity chromatography on Concanavalin A-Sepharose and molecular sieving. The enzyme thus obtained is practically free of any contaminating activities - endonuclease or phosphomonoesterase. The molecular weight of the exonuclease was determined (98 000 +/- 3 000 daltons) and some other parameters of the enzyme were calculated. The investigation of the pH and thermo-stabilities showed significantly narrow limits of the exonuclease activity. The effect of the urea on the enzyme activity has also been evaluated. PMID- 6289293 TI - Separation of SP1 alpha and SP1 beta on hydroxylapatite. PMID- 6289294 TI - [Oxidative-reductive processes in diabetes mellitus]. AB - Altogether 70 patients with diabetes mellitus of medium severity were examined to determine sulfhydryl (SH) groups, total glutathione (tG) and its individual forms, reduced (rG) and oxidized (oG), cytochrome oxidase activity (CCO), blood and urine V-O, and underoxidation coefficient of the urine with respect to the degree of hyperglycemia and glucosuria. Effect of sugar-reducing therapy on these parameters was studied in 26 subjects. A statistically significant reduction in SH groups balance, in oG content at the expense of rG, increase in CCO activity, in the blood and urine V-O, and in underoxidation coefficient of the urine were detected. The observed changes depend on the sugar content in the blood and urine, and do not disappear even when normoglycemia and aglycosuria are achieved after the use of sugar-reducing drugs. PMID- 6289296 TI - [Various problems in the pathogenesis of the early stage of leukemias]. PMID- 6289295 TI - [Immunoenzyme analysis. The comparative characteristics of 2 methods of obtaining a hormone and enzyme complex]. AB - Two methods for protein conjugation are routinely used in biochemical studies. One of them is based on the binding of free protein amino groups via functionally active elements of glutaraldehyde, whereas the other one makes use of covalent binding of proteins; carbohydrate groups of one of these proteins are preoxidized with sodium periodate to form free aldehyde groups. The possibility of employing these two methods for enzyme immunologic analysis of hypophyseal hormones was examplified by obtaining the complex of the ox growth hormone with horse radish peroxidase. PMID- 6289297 TI - Reduced availability of endogenously synthesized methionine for S adenosylmethionine formation in methionine-dependent cancer cells. AB - Methionine (Met) dependence--i.e., the inability of cultured cells to grow when Met is replaced by its immediate precursor homocysteine (Met-Hcy+ medium)--is a frequent component of the oncogenically transformed phenotype. Normal cells, on the other hand, grow in this medium. There have been reports [Hoffman, R. M. & Erbe, R. W. (1976) Proc. Natl. Acad. Sci. USA 73, 1523-1527; Hoffman, R. M., Jacobsen, S. J. & Erbe, R. W. (1978) Biochem. Biophys. Res. Commun. 82, 228-234] of normal or higher rats of Met biosynthesis in Met-dependent cells and a postulation that Met-dependent cells are deficient in utilization of endogenously synthesized Met as opposed to exogenously supplied Met. To answer the critical question of what biochemical reaction(s) requires preformed Met in Met-dependent cels, we labeled cells with Met-free [35S]Hcy or [35S]Met and determined the levels of Met, S-adenosylmethionine (AdoMet), and S-adenosylhomocysteine (AdoHcy). We report here experiments that demonstrate that Met-dependent cells synthesize a normal amount of endogenously synthesized Met and are deficient in utilizing this Met for AdoMet synthesis. In contrast, exogenously supplied Met is utilized normally for AdoMet biosynthesis. The ratio of AdoMet to AdoHcy is low in Met-dependent cells growing in Met-Hcy+ medium but is normal in Met+Hcy- medium. We determined that the low AdoMet/AdoHcy ratio probably limits growth of Met-dependent cells in Met-Hcy+ medium. PMID- 6289298 TI - Tissue-specific expression of mRNAs for dipeptidyl carboxypeptidase isoenzymes. AB - The molecular weight of newly synthesized dipeptidyl carboxypeptidase (angiotensin-converting enzyme; peptidyldipeptide hydrolase, EC 3.4.15.1) polypeptide primed in a reticulocyte lysate by poly(A)-containing RNA from mature rabbit testis was only about 65% that of the immunologically related species programmed by pulmonary RNA. Furthermore, in contrast to the pulmonary RNA dependent product, the synthesis of this testicular protein was not directed by RNA from testes of immature animals. These findings indicate that a shorter polypeptide chain and pubertal expression--the structural and regulatory properties that distinguish the testicular dipeptidyl carboxypeptidase isozyme- are determined pretranslationally. PMID- 6289299 TI - Control of transmembrane ion fluxes to select halorhodopsin-deficient and other energy-transduction mutants of Halobacterium halobium. AB - We describe a selection method for mutants altered in the generation and regulation of transmembrane ion flux in Halobacterium halobium. The method is based on experimental control of ion fluxes by a combination of light, ionophore, and external pH to generate an imbalance in the cells' proton circulation through their membranes. The steady-state proton circulation is increased by the introduction of a small inward proton leak with a protonophore. The cells are then illuminated to excite halorhodopsin, which hyperpolarizes the membrane and drives protons into the cells. As a result, wild-type cells suffer cytoplasmic acidification, which causes a dramatic loss of motility and suppresses their growth. These properties can be used to select for mutants that escape cytoplasmic acidification because either they lack halorhodopsin function or they have a greater capacity to eject protons during the illumination. In a popular selected by this method, 97% of the individual cells were demonstrably altered in ion flux properties. Cells were selected with alterations in the halobacterial rhodopsin, specifically with deficiencies in membrane potential generation by halorhodopsin and with increased cellular proton ejection by bacteriorhodopsin. We describe properties of one of the halorhodopsin-deficient strains, Flx37. PMID- 6289300 TI - Exocytosis regulates urinary acidification in turtle bladder by rapid insertion of H+ pumps into the luminal membrane. AB - Urinary acidification by the turtle bladder is due to a H+-ATPase that is located in the luminal membrane. The rate of H+ transport is stimulated by an increase in the ambient CO2. Using the fluorescent dye acridine orange, we showed that the mitochondria-rich cell of this equilibrium contains vesicles whose internal pH is acidic. We measured the pH of these vesicles by using endocytosed fluorescein isothiocyanate-labeled dextran and found it to be near 5.0. The pH increased after treatment with protonophores or metabolic inhibitors, suggesting that it was due to a H+ pump rather than to a Donnan effect. In bladders preloaded with fluorescent dextran, CO2 stimulated exocytosis and H+ transport measured simultaneously in the same bladder. The increase in the H+ current correlated well with the extent of exocytosis, and both were inhibited by pretreatment with colchicine. We conclude that the turtle bladder contains an intracellular reserve of vesicles containing H+ pumps and CO2 stimulates rapid fusion of these vesicles with the luminal membrane with consequent insertion of H+ pumps, thereby stimulating H+ secretion across the whole epithelium. PMID- 6289301 TI - Effects of adrenal cortical steroids and osmotic blood-brain barrier opening on methotrexate delivery to gliomas in the rodent: the factor of the blood-brain barrier. AB - The effect of adrenal cortical steroids and osmotic blood-brain barrier modification on methotrexate delivery to normal and glioma-bearing rats was studied. In animals with the avian sarcoma virus-induced glioma, osmotic blood brain barrier modification resulted in significantly increased delivery of methotrexate to the tumor-bearing hemisphere (including the tumor, the brain around the tumor, and the brain distant to the tumor), compared to the nonmodified hemisphere or to control animals. The administration of adrenal steroids, followed by intracarotid methotrexate, resulted in slightly decreased chemotherapeutic agent (methotrexate) delivery to the tumor, the brain around the tumor, and the brain distant to the tumor. When adrenal steroids were given prior to barrier modification and methotrexate therapy, the level of methotrexate was significantly less in the tumor. These studies provide evidence that the blood brain barrier exists in tumors and is a factor in drug delivery to tumors. Steroid administration greatly interferes with the enhancement of drug delivery to tumors that can be achieved with osmotic blood-brain barrier modification. PMID- 6289302 TI - Isolation of a human papillomavirus from a patient with epidermodysplasia verruciformis: presence of related viral DNA genomes in human urogenital tumors. AB - The DNA genome of a human papillomavirus (HPV), tentatively designated HPV-EV, was molecularly cloned from hand to leg lesions of a patient with epidermodysplasia verruciformis, a chronic skin disease associated with a 30% risk of developing cancer. Using stringent hybridization conditions, we observed less than 5% homology between HPV-EV and the cloned genomes of HPV-1, HPV-4, HPV 5, and HPV-5a. HPV-EV DNA showed approximately 6% homology with HPV-2 and 36% homology with HPV-3. These data suggest that HPV-EV is partially related to HPV 3. Using 32P-labeled cloned HPV-EV as probe in Southern blot hybridization experiments, we detected HPV-EV-related DNA in the carcinoma in situ (Bowenoid lesion) of the vulva of the patient from which HPV-EV was isolated. HPV-EV related DNA was detected in 2 of 10 vulva carcinomas and in 2 of 31 cervical carcinomas. Related DNA sequences were found in papillomas from each of two patients with condyloma acuminata (anogenital warts), which is of interest considering that condylomas have been reported to convert occasionally to carcinomas. The positive vulva DNAs were also probed with other cloned HPV DNAs: HPV-1, HPV-4, and HPV-5a-related sequences were not detected; HPV-3 and HPV-2 DNA probes detected strong and weak DNA bands, respectively, of the same size as found with HPV-EV. The HPV DNA sequences were present in the positive tumors mainly as free viral DNA molecules; no evidence for integration into cellular DNA was found. The emerging biological picture with papillomaviruses is that cells transformed by these viruses are maintained in a transformed state by free episomal genomes. Thus, our findings are consistent with the idea, but by no means establish, that HPVs play a role in human cancer by a similar mechanism. PMID- 6289303 TI - Electron microscopic visualization of the tetrodotoxin-binding protein from Electrophorus electricus. AB - Preparations of highly purified tetrodotoxin-binding protein (sodium channel) from the electric organ of the eel Electrophorus electricus were examined in negatively stained preparations. Structures observed in preparations exhibiting the highest tetrodotoxin binding tended to aggregate into ordered clusters with a unique ribbon-like conformation. The individual particles of these aggregates are elongated or rod-shaped, approximately 40 A wide and 170 A long. Stereoscopic imaging of the three-dimensional aspects of the structures revealed that the rod like image is not an edge view of a flattened disc but represents a cylindrical structure. Individual rods in nonclustered forms were also observed but with greater frequency in preparations with lower specific activity. The dimensions of the particles suggest that they represent a protein core formed by perhaps one copy of the large glycopeptide previously identified as being part of the sodium channel. The structure of the sodium channel component visualized by negative staining is discussed in the context of the excitable properties it contributes to biological membranes. PMID- 6289304 TI - Cointegrate formation by Tn5, but not transposition, is dependent on recA. AB - We have studied the effect of the recA-dependent homologous recombination system of Escherichia coli on both Tn5-mediated cointegrate formation and Tn5 transposition. We demonstrate here that, whereas transposition of Tn5 is independent of the recA gene product (as has been shown by other workers), Tn5 mediated cointegrate formation is strongly dependent on recA. The structures of both the simple transposition products and the cointegrates formed in a recA- background seem to be the same as those produced in a recA+ background. These results provide strong evidence that Tn5 does not transpose via an obligate cointegrate intermediate and suggest that the recA effect on cointegrate formation is exerted during the process of transposition. PMID- 6289305 TI - Adsorptive endocytosis of fibrin monomer by macrophages: evidence of a receptor for the amino terminus of the fibrin alpha chain. AB - Human fibrinogen and fibrin monomer were labeled with heme-octapeptide for cytochemical examination of their interaction with rabbit peritoneal macrophages in vitro. Upon short exposure to labeled fibrin monomer in solution with unlabeled fibrinogen, the cells became covered with surface-adsorbed monomer in nonaggregated form and having a characteristic trinodular shape. Within 30 min, the adsorbed monomer became fully internalized by vesicular uptake, with much of it being incorporated into lysosomal bodies. A concomitant loss of adsorptive capacity of the cell surface for uptake of more monomer accompanied the internalization. By contrast, labeled fibrinogen was not adsorbed by the macrophage surface. Some internalization of fibrinogen by passive fluid-phase pinocytosis was evident, but it was not accompanied by loss of absorptive capacity for fibrin monomer. The active uptake of monomer may have depended on binding to the amino terminus that is blocked by fibrinopeptide A in fibrinogen, because addition of synthetic peptide corresponding to the terminal Gly-Pro-Arg segment inhibited both the adsorption and the internalization of monomer. PMID- 6289306 TI - Labeling and selective recovery of newly synthesized viral DNA from simian virus 40-infected cells incubated with inorganic thiophosphate. AB - African green monkey CV-1 cells infected with simian virus 40 (SV40) were incubated in the presence of sodium [35S]thiophosphate. The viral DNA was prepared and the newly synthesized SV40 DNA molecules containing phosphorothioate groups were selectively recovered by affinity chromatography on organomercurial Sepharose columns. The thio-derivatized viral DNA occurred in both superhelical (form I) and relaxed circular (form II) configurations, and it gave the expected cleavage products upon digestion with the restriction endonucleases Kpn I and HindIII. Enzymatic digestion of 35S-labeled SV40 DNA to its component nucleotides and chromatographic separation of the resultant 5'-labeled phosphorothioate esters established the presence of the label in all four major nucleotides, but mainly in thymidylic acid. The ability to label and recovery selectively the newly synthesized polynucleotides by incubating cells with inorganic thiophosphate complements earlier findings [Sun, I. Y.-C. & Allfrey, V. G. (1982) J. Biol. Chem, 257, 1347--1353] that thiophosphate enters intracellular nucleotide pools and serves as an effective precursor for the phosphorylation of nuclear proteins in vivo. PMID- 6289307 TI - Site-directed psoralen crosslinking of DNA. AB - A technique has been developed for placing site-specific crosslinks into DNA by using a psoralen derivative containing a sulfhydryl group. Plasmid pBR322 was nicked at the unique BamHI restriction site, and mercurated pyrimidines were introduced by nick-translation. Then the psoralen was specifically directed to this site in the dark through a Hg-S linkage prior to irradiation to form a crosslink. The location of interstrand cross-links was examined by electron microscopy of Pst I-cut molecules and found to be at or near the BamHI site. PMID- 6289308 TI - In vitro synthesis of the first dipeptide of the beta subunit of Escherichia coli RNA polymerase. AB - Plasmids pNF1337 and pNF1341, which contain part of the L10 operon including the RNA polymerase beta-subunit gene, have been used as templates in vitro to investigate expression of the beta-subunit gene. For these studies, the synthesis of the first dipeptide of the beta subunit, fMet-Val, was measured instead of that of the entire protein. By using this dipeptide system, we studied the effects of RNA polymerase holoenzyme and L factor (nus A gene product) on fMET Val synthesis and compared the relative effects of the primary and secondary promoters in the L10 operon on expression of the beta-subunit gene. The results show that the inhibitory effect of RNA polymerase on beta-subunit synthesis and the stimulatory effect of L factor occur before formation of the first dipeptide bond. In this in vitro system, the secondary promoters account for about 50% of the total fMet-Val synthesized. Although the primary promoter is sensitive to guanosine 5'-diphosphate 3'-diphosphate in vitro, the secondary promoters are not affected by this nucleotide. PMID- 6289309 TI - Evidence for a spin-coupled binuclear iron unit at the active site of the purple acid phosphatase from beef spleen. AB - The purple acid phosphatase from beef spleen, which contains two iron atoms per molecule, is EPR silent in its native (oxidized) purple form. Treatment with mild reducing agents results in conversion to a pink, enzymatically active form, which exhibits an unusual EPR signal centered at g approximately equal to 1.77; double integration of the EPR spectrum gives one spin per two iron atoms. A similar EPR spectrum is observed for enzyme reduced anaerobically by one electron, using sodium dithionite. Variable-temperature magnetic susceptibility measurements show that the oxidized and reduced proteins are both antiferromagnetically coupled systems, with S = 0 and 1/2 ground states, respectively. Replacement of one of the iron atoms by zinc produces an FeZn enzyme with full catalytic activity. The FeZn enzyme exhibits a highly temperature dependent g = 4.3 EPR signal, and magnetic susceptibility data are consistent with an S = 5/2 paramagnet. Treatment of the FeZn enzyme with phosphate, a competitive inhibitor, results in sharpening of the EPR spectrum; double integration at 77 K gives one spin per iron. These results strongly suggest the presence of a spin-coupled bimetallic unit at the active site of the enzyme. PMID- 6289310 TI - Integration and expression of a truncated simian virus 40 early gene fragment in mammalian cells. AB - The recombinant plasmid p102 based on pBR322 carrying approximately equal to 50% of the replicator proximal early region of simian virus 40 (SV40) DNA, including the viral origin of replication, has been constructed. It lacks a major part of the large tumor (T) antigen 3'-coding region, the T-antigen termination codon, and the polyadenylylation site. The plasmid was transferred together with the herpes simplex virus thymidine kinase (TK) gene as a selectable marker to mouse LTK- cells. TK+ cell clones were isolated and their high molecular weight DNAs were shown by DNA blotting and hybridization experiments to contain the SV40 DNA fragment from the recombinant. In some of these clones, heterogeneous expression of the SV40 DNA fragment could be detected by immunofluorescence while, in control experiments in which a plasmid containing the complete SV40 early DNA region was used, this extensive heterogeneity of T-antigen expression was not observed. RNA . DNA hybridization experiments showed that the SV40-specific RNA of those clones is polyadenylylated. The molecular weight of the T-antigen related protein coded by p102 corresponded well to the expected coding capacity of the SV40 DNA fragment. Small tumor antigen was not expressed. PMID- 6289311 TI - A unique cell-surface protein phenotype distinguishes human small-cell from non small-cell lung cancer. AB - We have used radioiodination (125I) and two-dimensional polyacrylamide gel electrophoresis to determine that small- (oat) cell lung carcinoma (SCC)--a tumor with neuroendocrine features--possesses a surface protein pattern distinct from the other types of lung cancer cells (squamous, adeno-, and large-cell undifferentiated carcinoma). Twelve distinguishing proteins, 40 to 70 kilodaltons (kDal), characterized four separate lines of SCC; three of these, designated E (60 kDal; pI = 7.3), S (30 kDal; pI = 6.0), and U (57 kDal; pI = 5.6), may be unique SCC gene products and were identified only in [35S]methionine labeling of SCC and not in non-SCC or human fibroblasts. Two lines of adeno-, one of squamous, and one of undifferentiated large-cell lung carcinoma exhibited similar surface protein patterns to one another. Nine distinguishing proteins (40 to 100 kDal) and at least five large proteins (greater than 100 kDal) were unique to these lines. The surface protein phenotypes for SCC and non-SCC were distinct from those for human lymphoblastoid cells and fibroblasts. However, the neuroendocrine features of SCC were further substantiated because 6 of the 12 distinguishing SCC surface proteins, including E and U, were identified on human neuroblastoma cells. The proteins identified should (i) help define differentiation steps for normal and neoplastic bronchial epithelial cells, (ii) prove useful in better classifying lung cancers, and (iii) be instrumental in tracing formation of neuroendocrine cells. PMID- 6289312 TI - Mitochondrial DNA polymorphism in a maternal lineage of Holstein cows. AB - Two mitochondrial genotypes are shown to exist within one Holstein cow maternal lineage. They were detected by the appearance of an extra Hae III recognition site in one genotype. The nucleotide sequence of this region has been determined and the genotypes are distinguished by an adenine/guanine base transition which creates the new Hae III site. This point mutation occurs within an open reading frame at the third position of a glycine codon and therefore does not alter the amino acid sequence. The present pattern of genotypes within the lineage demands that multiple shifts between genotypes must have occurred within the past 20 years with the most rapid shift taking place in no more than 4 years and indicates that mitochondrial DNA polymorphism can occur between maternally related mammals. The process that gave rise to different genotypes in one lineage is clearly of fundamental importance in understanding intraspecific mitochondrial polymorphism and evolution in mammals. Several potential mechanisms for rapid mitochondrial DNA variation are discussed in light of these results. PMID- 6289313 TI - Chromosomal mapping of the simian sarcoma virus onc gene analogue in human cells. AB - The primate cell-derived transforming gene (v-sis) of simian sarcoma virus (SSV) is represented as a single copy marker within cellular DNAs of mammalian species including human. The human analogue of v-sis can be distinguished from its rodent counterparts by Southern blotting analysis of EcoRI-restricted DNAs. By testing for the presence of the human v-sis-related fragment, c-sis (human), in somatic cell hybrids possessing varying numbers of human chromosome, as well as in segregants of such hybrids, it was possible to assign c-sis to human chromosome 22. PMID- 6289314 TI - Structural basis for receptor binding heterogeneity of apolipoprotein E from type III hyperlipoproteinemic subjects. AB - The three major isoforms of human apolipoprotein E (apo-E2, -E3, and -E4) are coded for by three alleles (epsilon 2, epsilon 3, and epsilon 4) which have a common genetic locus. Previously, we demonstrated that E2, E3, and E4 differ in primary structure from one another at two substitution sites, site A (residue 112) and site B (residue 158). At sites A/B, apo-E2, -E3, and -E4 contain cysteine/cysteine, cysteine/arginine, and arginine/arginine, respectively. We demonstrated that the substitution of cysteine for arginine at site B is at least partly responsible for the defective binding of apo-E2 to human fibroblast low density lipoprotein receptors, compared to the normal binding activity of apo-E3 or -E4. Subjects with the genetic disorder type III hyperlipoproteinemia are phenotypically homozygous for apo-E2, but the binding activity of apo-E to the fibroblast receptor differs considerably from one type III individual to another. We therefore undertook a partial comparative sequence analysis of apo-E2 from three type III subjects whose apo-E displayed this heterogeneity. The subject with the poorest binding apo-E2 was genotypically homozygous for an apo-E allele (epsilon 2); cysteine was found at sites A and B. The subject with the most active apo-E2 was genotypically homozygous for an apo-E allele (epsilon 2); cystine was found at site A and at a new site (site C, residue 145). The epsilon 2 allele specifies a protein that has arginine at site B (residue 158); the epsilon 2 allele specifies a protein that has arginine at site C (residue 145). Therefore, the two alleles differ from one another by cysteine/arginine interchanges at two positions, sites B and C. The third subject, whose apo-E2 displayed binding activity intermediate between the activities of the other two, was genotypically heterozygous, having one epsilon 2 allele and one epsilon 2 allele. The intermediate binding activity of apo-E2 from this subject resulted from having a mixture of severely defective apo-E (specified by epsilon 2) and slightly defective apo-E (specified by epsilon 2). PMID- 6289315 TI - Chromosomal assignment of the human homologues of feline sarcoma virus and avian myeloblastosis virus onc genes. AB - Retroviral transforming genes, v-onc genes, are derived from normal cellular sequences that are called cellular onc (c-onc) genes. DNA from mouse-human somatic cell hybrids that have selectively lost human chromosomes was used in Southern blots to map the chromosomal location of two human onc genes. Cloned human homologues of retroviral onc genes were used as probes. Because the human c fes gene, which is homologous to feline sarcoma virus, segregates concordantly with human chromosome 15, and the human c-myb gene, which is homologous to avian myeloblastosis virus onc genes, segregates concordantly with human chromosome 6, we have assigned the c-fes and the c-myb genes to human chromosomes 15 and 6, respectively. Nonrandom chromosomal defects involving these human chromosomes have been observed in neoplasms. These studies should be valuable in determining whether specific rearrangements involving these chromosomes result in the abnormal expression of these onc genes in human malignancies. PMID- 6289316 TI - Transmembrane channel formation by complement: functional analysis of the number of C5b6, C7, C8, and C9 molecules required for a single channel. AB - Earlier studies have shown that sequential treatment of resealed erythrocyte ghosts with C5b6, C7, C8, and C9 leads to insertion of hydrophobic peptides from these complement proteins into the membrane and assembly of transmembrane channels. The number of molecules of each of the proteins required for assembly of the membrane-associated channel structure was evaluated by measuring the quantitative relationship between the doses of the individual proteins and the release of two trapped markers, sucrose and inulin, from ghosts after channel formation. The incubation period was sufficient to attain equilibrium of marker distribution between the ghosts and the extracellular fluid. Two markers of different size (sucrose and inulin, 0.9 and 3 nm molecular diameter, respectively) were used in order to develop information on the molecular composition of small and large channels, respectively. We found that participation of C5b6, C7, and C8 in channel formation displayed one-hit characteristics, regardless of marker size. By contrast, the participation of C9 was one-hit with respect to the sucrose marker, whereas with respect to the inulin marker the C9 reaction was multi-hit. Our results are compatible with the view that these markers are released through a channel structure in the membrane that is a monomer of C5b--9 of the composition C5b61 C71C81C9n, in which n = 1 for channels permitting passage of sucrose and n = 2 for channels allowing transit of inulin. PMID- 6289317 TI - IgG2a monoclonal antibodies inhibit human tumor growth through interaction with effector cells. AB - Monoclonal antibodies of IgG2a isotype specifically inhibited growth of human tumors in nude mice. Twenty-three monoclonal antibodies of other isotypes showed no tumoricidal reactivity. Complement depletion of nude mice had no effect on tumor suppression by monoclonal antibody. The role of T and killer cells as mediators of the monoclonal antibody effect in nude mice was virtually excluded. On the other hand, macrophages were strongly incriminated as effector cells because silica treatment of nude mice abolished the tumoricidal effect of monoclonal antibody. IgG2a monoclonal antibody-dependent macrophage-mediated cytotoxicity assays with human tumor cells in culture resulted in specific lysis of tumor cells. PMID- 6289318 TI - Dimer-size DNA circles in a leukemic cell immortalized with the Epstein-Barr virus. AB - The intracellular state of the 30 viral genome equivalents of Epstein-Barr virus (EBV) DNA carried in latent form by the CII cell line, established from a chronic lymphocytic leukemia patient, has been partially characterized. The CII line, which has markers confirming its tumor origin, extends the analysis of the intracellular state of EBV DNA to include other, non-Burkitt lymphoid tumor cells. Monomer-size, free, circular EBV genomes, the major intracellular viral DNA species in other EBV-transformed cells, were absent or present in only minor amounts. Instead, EBV DNA sequences were found associated with a circular DNA form twice the size of the 110 x 10(6) Mr EBV genome. Though circular dimers of mtDNA have been found exclusively in human leukemic lymphocytes, the CII line is similar to normal cells in having only monomer-size mtDNA molecules, which can occur either singly or as catenated forms of two or more interlocking 5 micrometer mtDNA circles. PMID- 6289319 TI - Healthy carriers of a human retrovirus, adult T-cell leukemia virus (ATLV): demonstration by clonal culture of ATLV-carrying T cells from peripheral blood. AB - Adult T-cell leukemia virus (ATLV) or ATLV-associated antigen (ATLA)-positive cell clones were isolated from peripheral blood lymphocytes of all five anti-ATLA seropositive healthy adults tested by a limiting-dilution culture method in the presence of T-cell growth factor (TCGF) but from none of six seronegative adults similarly tested. Although ATLA-positive cells were not always detected in mass cultures of the seropositive lymphocytes, they were found consistently in T-cell cloned cultures of those lymphocytes. Continuous cultures of the ATLA-positive clones obtained were dependent on TCGF. Five clones derived from each of the five donors were maintained for greater than 4 months and, during culture, all of them acquired the ability to grow without added TCGF. The ATLA-positive clones formed rosettes with erythrocytes and expressed Leu 1, Leu 3a, and Leu 4 antigens but not Leu 2a antigen. These results indicate that anti-ATLA-seropositive healthy individuals carry ATLV in T cells circulating in their peripheral blood. PMID- 6289320 TI - Human genome contains four genes homologous to transforming genes of Harvey and Kirsten murine sarcoma viruses. AB - Harvey and Kirsten murine sarcoma viruses each encode a structurally and functionally related 21-kilodalton protein (p21), which is the transforming protein of each virus. Using probes from the 0.9-kilobase (kb) p21-coding region of each virus (called v-Ha-ras and v-Ki-ras, respectively), we have molecularly cloned from normal human genomic DNA the sequences that hybridize to these probes. Four evolutionarily divergent restriction endonuclease fragments were isolated. Two hybridized preferentially to v-Ha-ras and were designated human c Ha-ras1 and c-Ha-ras2; two hybridized preferentially to v-Ki-ras and were called c-Ki-ras1 and c-Ki-ras2. Human c-Ha-ras1 contained 0.9 kb of sequence homologous with v-Ha-ras interspersed with three intervening sequences; this gene was closely related to a previously cloned rat c-Ha-ras gene that also contained intervening sequences. Human c-Ha-ras2 was more divergent from v-Ha-ras and also hybridized poorly to human c-Ha-ras1. One c-Ki-ras gene contained 0.9 kb homologous to v-Ki-ras and had one intervening sequence, whereas the other contained only 0.3 kb homologous to v-Ki-ras. The results indicated that human DNA contains several copies of the c-ras family and that c-Ha-ras1 (with intervening sequences) was more highly conserved evolutionarily than was c-Ha ras2. PMID- 6289321 TI - Inhibition of hepatic cholesterol synthesis in mice by sterols with shortened and stereochemically varied side chains. AB - Mice were fed cholesterol or various other sterols for 26 hr, after which the amount of hepatic cholesterol synthesis was measured in a cell-free system. The following sterols were as effective as cholesterol itself in depressing the conversion of acetate into sterol: pregn-5-en-3 beta-ol, which lacks an isohexyl group on C-20; (E)-17(20)-dehydrocholesterol, in which the isohexyl group is fixed to the right; (E)-20(22)-dehydrocholesterol, in which C-23 is oriented away from the nucleus; and 20-epicholesterol. Moreover, when the isohexyl group was fixed to the left in (Z)-17(20)-dehydrocholesterol, this dietary sterol, identified in the liver, caused not only a depression in the conversion of both mevalonate and squalene into sterols. The incorporation of acetate into fatty acids was not depressed, nor did the (Z)-sterol appear to have a generalized effect on membranous enzymes, because the activity of glucose-6-phosphatase was unaffected. Thus, feedback inhibition was retained when the stereochemistry of cholesterol's side chain was drastically changed and even after the nearly complete removal of the side chain. This implies that the side chain is only minimally recognized by the mechanisms involved in feedback inhibition. PMID- 6289322 TI - Aluminum: a requirement for activation of the regulatory component of adenylate cyclase by fluoride. AB - Activation of the purified guanine nucleotide-binding regulatory component (G/F) of adenylate cyclase by F- requires the presence of Mg2+ and another factor. This factor, which contaminates commercial preparations of various nucleotides and disposable glass test tubes, has been identified as Al3+. In the presence of 10 mM Mg2+ and 5 mM F-, AlCl3 causes activation of G/F with an apparent activation constant of approximately 1-5 muM. The requirement for Al3+ is highly specific; of 28 other metals tested, only Be2+ promoted activation of G/F by F-. PMID- 6289323 TI - Differentiation between alpha promoter and regulator regions of herpes simplex virus 1: the functional domains and sequence of a movable alpha regulator. AB - The herpes simplex virus genome consists of at least three groups of genes- alpha, beta, and gamma--whose expression is coordinately regulated and sequentially ordered in a cascade fashion. We have established that the elements involved in regulation of alpha genes are a sequence that promotes gene expression and a sequence that confers alpha regulation on the gene by responding to trans-acting regulatory signals. The domains of these sequences were mapped by determining the regulation of thymidine kinase (TK) in L cells converted to TK+ phenotype by chimeric TK indicator genes. The chimeric genes were constructed from appropriate portions of the TK gene fused to donor sequences derived from the 5' nontranscribed and nontranslated leader portions of the viral alpha gene 4. The results were as follows. (i) The natural beta TK indicator extending 5' up to -80 and the chimeric alpha TK extending 5' up to -110 both converted cells to TK+ phenotype but were not regulated. (ii) A segment of the regulator region of the alpha gene 4, extending 5' from position -110, confers inducible alpha-type regulation when fused to the nonregulated but expressible beta TK indicator described above. (iii) The extent of gene induction appears to hinge on the size of the regulatory region inserted into the chimeric gene and correlates with the presence of repeated consensus sequences and G+C-rich inverted repeats in the regulatory region of the alpha gene 4 and other alpha genes. PMID- 6289324 TI - Construction of poxviruses as cloning vectors: insertion of the thymidine kinase gene from herpes simplex virus into the DNA of infectious vaccinia virus. AB - We have constructed recombinant vaccinia viruses containing the thymidine kinase gene from herpes simplex virus. The gene was inserted into the genome of a variant of vaccinia virus that had undergone spontaneous deletion as well as into the 120-megadalton genome of the large prototypic vaccinia variant. This was accomplished via in vivo recombination by cotransfection of eukaryotic tissue culture cells with cloned BamHI-digested thymidine kinase gene from herpes simplex virus containing flanking vaccinia virus DNA sequences and infectious rescuing vaccinia virus. Pure populations of the recombinant viruses were obtained by replica filter techniques or by growth of the recombinant virus in biochemically selective medium. The herpes simplex virus thymidine kinase gene, as an insert in vaccinia virus, is transcribed in vivo and in vitro, and the fidelity of in vivo transcription into a functional gene product was detected by the phosphorylation of 5-[125I]iodo-2'-deoxycytidine. PMID- 6289325 TI - Nucleotide sequences involved in bacteriophage T4 gene 32 translational self regulation. AB - We have determined the nucleotide sequence of a cloned segment of the bacteriophage T4D chromosome, which contains the regulatory sequences and the structural gene (gene 32) for the single-stranded DNA binding protein (gp32). The amino acid sequence predicted by translation of the structural gene agrees well with that published for gp32 [Williams, K. R., Lo-Presti, M. B., Setoguchi, M. & Konigsberg, W. H. (1980) Proc. Natl. Acad. Sci. USA 77, 4614-4617]. To localize the nucleotide sequence involved in translational self-regulation of gene 32, we have constructed a series of plasmids in which gene 32 is fused to an amino terminal deletion mutant of the beta-galactosidase gene of Escherichia coli. Expression of a beta-galactosidase fusion protein that contains only the first seven amino acids of gp32 is still repressed by gp32. The ribosomal binding site of gene 32 is flanked by a repetitive A+T-rich sequence. Preferential and cooperative binding of gp32 to this region of its mRNA could inhibit translation initiation and, thus, would account for the autoregulation. PMID- 6289326 TI - Activation of early adenovirus transcription by the herpesvirus immediate early gene: evidence for a common cellular control factor. AB - Adenovirus mutants carrying a defective E1A gene, such as dl312, are unable to express any of the early viral genes upon infection of HeLa cells. However, efficient expression of the other early adenovirus genes was obtained when dl312 infected HeLa cells were coinfected with pseudorabies virus, a herpesvirus. By employing a temperature-sensitive pseudorabies mutant (tsG1) it was demonstrated that the herpesvirus function responsible for the induction of adenovirus transcription was the immediate early gene, a gene required for the activation of herpesvirus early gene expression and the maintenance of early and late herpesvirus transcription. Specifically, HeLa cells coinfected with dl312 and tsG1, when shifted to the nonpermissive temperature, lost their capacity to express the early adenovirus genes. Furthermore, activation of early adenovirus gene expression in herpesvirus coinfection occurred earlier and at a higher level than in wild-type adenovirus infection. Therefore, the herpesvirus immediate early protein not only activates the early adenovirus transcription units but apparently does so more efficiently than the adenovirus E1A gene product. Because of this fact, we argue that the activation, either by the E1A protein or the herpesvirus immediately early protein, most likely occurs indirectly through interaction with a cellular protein rather than by a direct recognition of regulatory sequences at the adenovirus promoters. PMID- 6289327 TI - Prostaglandins E1 and E2 interact with prostaglandin F2alpha to regulate initiation of DNA replication and cell division in swiss 3T3 cells. AB - Prostaglandin (PG) E1 or E2 added at 2-1,000 ng/ml to quiescent cultures of Swiss 3T3 cells synergistically enhanced the rate of initiation of DNA replication stimulated by PGF2 alpha alone or with insulin. Neither PGD2 nor PGF1 alpha had any effect with PGF2 alpha. An increase in the rate of entry into S phase also occurred when PGE1 or PGE2 was added 8 or 15 hr after addition of PGF2 alpha. However, adding PGE1 and PGE2 together with PGF2 alpha did not further enhance the synergistic effect observed with PGE1 or PGE2 separately. The synergistic effect was also observed in stimulation of 2-deoxyglucose uptake but not in early changes of intracellular levels of cAMP. These results may be relevant in understanding the control of fibroblastic proliferation in wound healing and may provide an alternative mechanism for oncogenic transformation. PMID- 6289328 TI - Transcriptional activity of the transposable element Tn10 in the Salmonella typhimurium ilvGEDA operon. AB - Polarity of Tn10 insertion mutations in the Salmonella typhimurium ilvGEDA operon depends on both the location and the orientation of the Tn10 element. One orientation of Tn10 insertions in ilvG and ilvE permits low-level expression of the downstream ilvEDA and ilvDA genes, respectively. Our analysis of Salmonella ilv recombinant plasmids shows that this residual ilv expression must result from Tn10-directed transcription and does not reflect the presence of internal promoters in the ilvGEDA operon, as was previously suggested. The opposite orientation of Tn10 insertion in ilvE prevents ilvDA expression, indicating that only one end of Tn10 is normally active in transcribing adjacent genes. Both orientations of Tn10 insertion in ilvD exert absolute polarity on ilvA expression. Expression of ilvA is known to be dependent on effective translation of ilvD, perhaps reflecting the lack of a ribosome binding site proximal to the ilvA sequence. Therefore, recognition of the ability of Tn10 to promote transcription of contiguous genes in the ilvGEDA operon apparently requires the presence of associated ribosome binding sites. PMID- 6289329 TI - Transposon Tn10 provides a promoter for transcription of adjacent sequences. AB - Promoters located within the Tn10 insertion element cause transcription of "host" sequences adjacent to both ends of the inserted Tn10 element. These promoters are usually not observed in genetic experiments because their transcripts are efficiently terminated at nearby rho-dependent termination sites. The observations presented here provide an explanation for several confusing aspects of transposon behavior and suggest the possibility that many transposons possess promoters that have escaped detection for similar reasons. PMID- 6289330 TI - Identity of human epidermal growth factor (EGF) receptor with glycoprotein SA-7: evidence for differential phosphorylation of the two components of the EGF receptor from A431 cells. AB - A 165-kilodalton (kDal) surface glycoprotein encoded by human chromosome 7 (SA-7) had been characterized by using antisera raised against human chromosome 7 containing somatic cell hybrids. We now present evidence that SA-7 is the human receptor for epidermal growth factor (EGF) and that these antisera recognize human-specific determinants. The gene coding for the human EGF receptor is localized to the p12 to p22 region of chromosome 7. We have characterized the 145 kDal/165-kDal EGF receptor doublet of A431 cells after immunoprecipitation of radiolabeled cell extracts with these antisera. We find that a protein with endogenous kinase activity copurifies with the A431 receptor doublet and that both components of the doublet contain phosphotyrosine and phosphothreonine and the 165-kDal component contains phosphoserine as well. Further, although each component of the receptor doublet has an average pI of 7, both display charge heterogeneity and appear to have unique charge isomers. The relationship between the two components of the A431 EGF receptor is discussed. PMID- 6289331 TI - Isolation of 16L virus: a rapidly transforming sarcoma virus from an avian leukosis virus-induced sarcoma. AB - We have isolated a replication-defective rapidly transforming sarcoma virus (designated 16L virus) from a fibro-sarcoma in a chicken infected with td107A, a transformation-defective deletion mutant of subgroup A Schmidt-Ruppin Rous sarcoma virus. 16L virus transforms fibroblasts and causes sarcomas in infected chickens within 2 wk. Its genomic RNA is 6.0 kilobases and contains sequences homologous to the transforming gene (fps) of Fujinami sarcoma virus (FSV). RNase T1 oligonucleotide analysis shows that the 5' and 3' terminal sequences of 16L virus are indistinguishable from (and presumably derived from) td107A RNA. The central part of 16L viral RNA consists of fps-related sequences. These oligonucleotides fall into four classes: (i) oligonucleotides common to the putative transforming regions of FSV and another fps-containing avian sarcoma virus, UR1; (ii) an oligonucleotide also present in FSV but not in UR1; (iii) an oligonucleotide also present in UR1 but not in FSV; and (iv) an oligonucleotide not present in either FSV, UR1, or td107A. Cells infected with 16L virus synthesize a protein of Mr 142,000 that is immunoprecipitated with anti-gag antiserum. This protein has protein kinase activity. These results suggest that 16L virus arose by recombination between td107A and the cellular fps gene. PMID- 6289332 TI - Evidence from in vitro murine immunologic assays that some phenolic food additives may function as antipromoters by lowering intracellular cyclic GMP levels. PMID- 6289333 TI - Neonatal herpes simplex virus infection in guinea pigs. PMID- 6289334 TI - Receptor binding affinities and biological activities of angiotensin analogues in the rat uterus. PMID- 6289335 TI - Depressant effects of drugs used in myotonia therapy. PMID- 6289336 TI - An alpha-adrenergic component of reflex bronchoconstriction induced by unilateral, orthodromic stimulation of the carotid sinus nerves. PMID- 6289337 TI - Cytochrome P450 dependent monooxygenases in nasal epithelial membranes: effect of phenobarbital and benzo(A)pyrene. PMID- 6289338 TI - Urotensin I - a novel CRF-like peptide in Catostomus commersoni urophysis. PMID- 6289339 TI - Alpha adrenoceptor mediated responses in aorta from three month streptozotocin diabetic rats. PMID- 6289340 TI - Programming of rat hepatic arylhydrocarbon hydroxylase activity during late neonatal period. PMID- 6289341 TI - Regulation of cytoplasmic Ca++ by smooth muscle membranes. PMID- 6289342 TI - Effects of verapamil with calcium chloride and calcium chloride plus A23187 on rat cerebellar levels of cyclic GMP. PMID- 6289344 TI - Clonidine and carbachol inhibit neurotransmission to mesenteric arteries in the guinea pig. PMID- 6289343 TI - The role of alpha adrenoreceptor subtypes in the asymmetric response of the rabbit ear artery to intimal or adventitial amines. PMID- 6289345 TI - Effects of sodium substitutes on ouabain induced transient inward current. PMID- 6289346 TI - 3H-(3MeHis2)thyrotropin releasing hormone receptors in rat central nervous system. PMID- 6289347 TI - Differential block of A beta, A delta and C fibers with ouabain. PMID- 6289348 TI - Postnatal development of the norepinephrine stimulated cyclic AMP response in rat spinal cord. PMID- 6289349 TI - The role of the coxal gland in ionic, osmotic, and pH regulation in the horseshoe crab Limulus polyphemus. PMID- 6289350 TI - The nature of the binuclear copper site in Limulus and other hemocyanins. PMID- 6289351 TI - Chloride and pH dependence of cooperative interactions in Limulus polyphemus hemocyanin. PMID- 6289352 TI - Transplantation of the human insulin gene into fertilized mouse eggs. PMID- 6289353 TI - Cell-cell contact, cyclic AMP, and gene expression during differentiation of the cellular slime mold Dictyostelium discoideum. PMID- 6289354 TI - Studies on the chalcone synthase gene of two higher plants: petroselinum hortense and matthiola incana. AB - Two higher plant systems are presented which allow to study coordinated gene expression of the light-induced metabolic pathway of flavonoid biosynthesis: tissue culture cells of Petroselinum hortense (Apiaceae) and different developmental stages of various genotypes of Matthiola incana (Brassicaceae). The gene structure of the chalcone synthase is mainly studied. A cDNA clone (pLF56) of parsley has been constructed and characterized conferring the chalcone synthase gene sequence. Strong cross hybridization between the parsley cDNA and Matthiola DNA allowed to identify a HindIII fragment (6000 bp) identical in size for parsley and different Matthiola wild type lines and a mutant line. PMID- 6289356 TI - Apparent influence of adrenocorticotrophic hormone (ACTH) in the USP rabbit pyrogen test and evaluation of LAL as an alternative procedure. PMID- 6289355 TI - Detection of Limulus amebocyte lysate reactive material in capillary flow hemodialyzers. PMID- 6289357 TI - Inherited disorders of hormone resistance. PMID- 6289358 TI - Phenotypic manifestations of Gaucher disease: clinical features in 48 biochemically verified type 1 patients and comment on type 2 patients. PMID- 6289359 TI - Human brain receptor alterations in suicide victims. AB - A comparison was made of human postmortem muscarinic-cholinergic, beta-adrenergic and serotonergic (presynaptic) recognition sites in cortical tissues derived from suicide and homicide (control) victims. An elevation of 47% and 35% in the suicide group compared to controls was observed in receptor ligand binding for 3H quinuclidinyl benzilate (QNB, muscarinic antagonist) and 3H-imipramine (IMI, a presynaptic serotonin marker), respectively. In contrast, no appreciable differences in 3H-dihydroalprenolol (DHA, beta-adrenergic antagonist) binding were observed between the two groups. Additionally, tissues from both groups of subjects were analyzed for tricyclic antidepressive agent (TAD) content. High performance liquid chromatographic (HPLC) tissue analysis revealed no detectable levels of tricyclic agents with an assay sensitivity of 50 picograms/mg tissue. The results presented herein demonstrate neurotransmitter-receptor alterations in suicide subjects compared to homicide (control) victims. The attendant roles of serotonergic and muscarinic-cholinergic processes in the psychobiology of suicide and depression are addressed. PMID- 6289360 TI - Regional glucose metabolism in mouse brain following ACTH peptides and naloxone. AB - Following intracerebroventricular (ICV) injection of ACTH1-24 significant decreases in 2-deoxyglucose (2DG) uptake were observed in frontal cortex and pyriform cortex, and an increase in thalamus. No such changes were observed following ICV MSH/ACTH4-10. Regional changes in 2DG uptake in olfactory bulb, pyriform cortex, thalamus and cerebellum were significantly correlated with the excessive grooming induced by ACTH1-24. Grooming behavior not induced by ACTH1-24 was not correlated with 2DG changes in any of these regions. Naloxone treatment did not significantly alter the regional pattern of 2DG uptake. In naloxone pretreated mice ACTH1-24 did not induce significant changes in regional 2DG uptake. Following a series of footshocks, 2DG uptake increased in the hypothalamus, tectum and hippocampus. This pattern of changes is different from that observed following ICV ACTH1-24 and cannot therefore be attributed to ACTH secretion during the stress. PMID- 6289361 TI - 3H-Codeine binding in the guinea pig lower brain stem. AB - Saturable binding of (-)-3H codeine was found in the guinea pig medulla (KD = 5.6 x 10(-7) M, Bmax = 1.4 pmol/mg protein), whereas little stereospecific binding was detected (KD = 4.4 x 10(-5) M). The saturable binding of (-)-3H codeine was slightly enhanced by Na+ and by Mg++ but not by Li++ and Ca++. The enhancement appears to be due to an increase in the number of receptor sites. (-)-3H-Codeine binding was displaced by (-)- and (+)-codeine, morphine, (-)- and (+)-methadone but not by barbiturates. Naloxone, at a high concentration (1 x 10(-5) M), inhibits the binding by only 40%. This agrees with our previously published data which shows that the optical isomers of codeine had significant antitussive effects in the cat, these effects not being antagonized by naloxone. A class of opiate antitussive receptors, which are less naloxone-sensitive and less stereoselective than the mu receptors, is implicated. PMID- 6289362 TI - Effects of oral atenolol and propranolol on blood pressure, heart rate and plasma cyclic adenosine 3':5'-monophosphate in borderline hypertensive patients. AB - The effects of oral treatment with atenolol and propranolol on blood pressure, heart rate and plasma cyclic adenosine 3':5'-monophosphate (cAMP) were studied in a group of young borderline hypertensives. Observations were accomplished after a period of recumbent relaxation and also following physiological stress testing. The latter significantly increased blood pressure, heart rate and plasma cAMP. A direct relationship was found between heart rate and plasma cAMP at rest and also following psychological stress. This relationship was lost following drug treatments. Both beta-blockers significantly reduced all measured variables in both conditions. However, plasma cAMP was most affected by propranolol. PMID- 6289363 TI - Neural plasticity. Part 4. Lesion-induced reorganization of the CNS: recovery phenomena. AB - Most patients treated by physical therapists have suffered some neurological trauma resulting from disease or injury. The traditional teaching used to be that damage of central neurons is irreversible. Within the last decade, however, it has been necessary to cast aside this traditional view because of accumulating evidence that the brain is endowed with remarkable plasticity. This paper reviews experimental evidence revealing morphological and functional changes occurring in the CNS in response to neural lesions. Morphological responses to injury include collateral and terminal sprouting, retrieval of vacated synapses, alterations in the ultrastructure of surviving synapses, and denervation supersensitivity. Functional and adaptive changes induced by injury include the unmasking of ineffective synapses, shifts in receptive fields, and reorganization or altered effectiveness of surviving neural networks. These recovery phenomena attest to the brain's dynamic properties. These new insights contradict our conventional view of the absence of growth and reorganizational capabilities in CNS neurons. These newly identified "recovery phenomena" are destined to have a significant impact on physical therapy in the future. PMID- 6289364 TI - Applications of flash spectroscopy to molecules of biological relevance. PMID- 6289365 TI - Resolved fluorescence emission spectra of iron-free cytochrome c. PMID- 6289366 TI - Photochemical reactions of dibromothymoquinone. PMID- 6289367 TI - Agonistic behaviour in rats: evidence for non-involvement of opioid mechanisms. AB - It has recently been proposed that a stress-activated, endogenous analgesia mechanism would be adaptive in situations in which pain perception might otherwise disrupt effective behavioural performance. In a semi-natural test situation, the current study examined two predictions arising from this hypothesis: (1) in a manner analogous to other stressors, agonistic experience should produce analgesia and, if naloxone-sensitive opioid mechanisms are implicated, then (2) pretreatment with naloxone should block the development of this response and alter the displayed behaviour patterns. Neither prediction was substantiated by the data. Experience of an agonistic encounter failed to produce analgesia in either resident or intruder animals. Furthermore, naloxone hydrochloride (1-25 mg/kg) was also without effect on patterns of offense or defense. Data are discussed in relation to the critical nature of the stimulus factors involved in the activation of endogenous analgesic mechanisms and the postulated involvement of such mechanisms in biologically-adaptive behaviours. PMID- 6289368 TI - [Care of patients after attempted suicide: tasks, problems and possibilities of improvement]. AB - Inpatient and post-inpatient psychiatric/psychotherapeutic treatment of suicides poses a considerable number of problems which have to be handled by the institutions concerned: These problems are formidable because of the high incidence of attempted suicides and the considerable amount of work necessarily involved in caring for such persons. Care should by no means be restricted to the acute crisis; attempts should also be directed by preventing relapses and at improving the overall psychopathological status and social readjustment. The present manner in which suicides are looked after has quite a number of drawbacks, e.g. insufficient attention is paid to the requirements of the inpatient; patients fail to refer to the hospital during the period of their post suicide attempt care; the incidence of "dropouts" during outpatient aftercare is very high; there is insufficient cooperation between the persons and institutions involved in the care programme. Improvements are presented which have been successfully tried out in practice. PMID- 6289369 TI - Strain difference in morphine-induced increase in plasma cyclic AMP and cyclic GMP levels in relation to locomotor activity in male mice. AB - The effects of morphine on plasma cyclic nucleotide levels and on locomotor activity were investigated in four inbred strains of male mice. Morphine increased both cyclic AMP and cyclic GMP levels as well as locomotor activity in C57BL/6N mice. In BALB/cAnN mice, morphine increased plasma cyclic GMP levels and motor activity without changes in plasma cyclic AMP levels. In C3H/HeN mice, morphine increased plasma cyclic GMP levels without changing cyclic AMP levels and motor activity, but neither plasma cyclic nucleotide levels nor motor activity were increased by morphine in DBA/2N mice. Epinephrine and carbachol increased plasma cyclic AMP and cyclic GMP levels, respectively, in both C57BL and DBA mice. These results show that there is a significant strain difference in the effects of morphine on plasma cyclic nucleotide levels as well as motor activity. The major cause of strain difference in the effects of morphine on cyclic nucleotide levels is unlikely to be due to the difference in the regulation of adenylate cyclase linked to adrenoceptors or that of guanylate cyclase connected with cholinoceptors. PMID- 6289370 TI - Learning impairment in the radial-arm maze following prolonged cannabis treatment in rats. AB - Chronic oral administration of cannabis extract to rats (daily delta 9 tetrahydrocannabinol dose 20 mg/kg) was examined in three experiments for its residual effect on radial-arm maze learning following a 1-month drug-free period. Learning a simple eight-arm maze was significantly impaired in rats treated for either 6 months (Experiment I) or 3 months (Experiment II) with the drug. In Experiment III, animals that received the extract for 3 months exhibited significant learning deficits on a much more difficult 12-arm radial maze. The results demonstrate that the deleterious effects of cannabis on radial-arm maze learning are probably due to a tendency toward increased vigilance and perseveration, possibly combined with an impaired utilization of spatial cues. PMID- 6289371 TI - Age response of EUE cells exposed to 31-MeV protons. PMID- 6289372 TI - [Dynamics of the content and aspects of the metabolism of nicotinamide coenzymes in irradiated animals]. PMID- 6289374 TI - [Diagnosis of insulinoma of the pancreas by computed tomography ]. AB - By example of a case the possibility for diagnosis of insulinoma by sequence computed tomography is demonstrated. If clinical symptoms are clear computed tomography should be performed before angiography, as in most cases sonography is not very successful for localisation. Because of high vascularisation of the tumor it can be found by computed tomography after one or more intravenous bolus injections or after high dose infusion of a renal excreted contrast medium. By aid of magnification technique and sequence computed tomography insulinomas of 5 mm in diameter can be recognized. PMID- 6289373 TI - [Tumors around the jugular foramen--diagnosis and clinical findings ]. PMID- 6289375 TI - Work in progress: 252Cf neutron brachytherapy for hemispheric malignant glioma. AB - It has been shown that 252Cf neutron brachytherapy of hemispheric malignant glioma can be readily carried out and may be combined with external, whole brain photon beam therapy of 6,000 rad (60 Gy) administered over a five-week to seven week period. The ten patients who were studied tolerated both procedures well. There was improvement in performance status, and a decrease in tumor size was observed upon computed tomographic scanning. PMID- 6289376 TI - Three-dimensional structure determination by electron microscopy of two dimensional crystals. PMID- 6289377 TI - [ATP synthesis and H+-translocation in H+-ATP synthetase (Fo.F1)]. PMID- 6289379 TI - Neutrophil aggregating and chemokinetic properties of a 5,12,20-trihydroxy 6,8,10,14-eicosatetraenoic acid isolated from human leukocytes. AB - Suspensions of human peripheral blood leukocytes were incubated with arachidonic acid and calcium ionophore A23187. Acidic lipid extracts were purified by silicic acid chromatography and reversed phase high performance liquid chromatography. A polar metabolite with an ultraviolet absorption spectrum identical to that of leukotriene B4 was identified by gas chromatography and mass spectrometry as a 5,12,20-trihydroxy-6,8,10,14-eicosatetraenoic acid. The data indicated that this compound was the 20-hydroxy metabolite of leukotriene B4. The compound showed chemokinetic activity towards human polymorphonuclear leukocytes and aggregating activity towards rat peritoneal polymorphonuclear leukocytes. Its activity was greater than that of 5-hydroxy-6,8,11,14-eicosatetraenoic acid but less than that of leukotriene B4. It is concluded that the 15,12,20-trihydroxyeicosatetraenoic acid may play a role as a mediator of inflammatory reactions. PMID- 6289378 TI - [Proton-pump mechanism in bacteriorhodopsin]. PMID- 6289381 TI - Studies on the mechanism of formation of the 5S, 12S-dihydroxy-6,8,10,14(E,Z,E,Z) icosatetraenoic acid in leukocytes. AB - Incubation of peripheral blood leukocytes with arachidonic acid (and ionophore A23187) led to the formation of leukotriene B4, delta 6-trans-leukotriene B4, delta 6-trans-12-epi-leukotriene B4, 5-hydroxy-icosatetraenoic acid, 12-hydroxy icosatetraenoic acid and of 5S,12S-dihydroxy-6,8,10,14-(E,Z,E,Z)-icosatetraenoic acid (5S,12S-DiHETE). Incubation of leukocytes with leukotriene A4 resulted in the formation of leukotriene B4 and of its two delta 6-trans-isomers but not of the 5S, 12S-DiHETE. 18O2 labeling experiments have shown that the hydroxyl groups at C5 and C12 in the 5S,12S-DiHETE are derived from molecular oxygen. The tetraacetylenic analog of arachidonic acid was found to be potent inhibitor of the formation of the 5S,12S-DiHETE whereas it potentiated the synthesis of the 5 hydroxy acid and of leukotriene B4. Addition of the 12-hydroxy-icosatetraenoic acid to leukocytes, or of the 5-hydroxy-icosatetraenoic acid to a suspension of platelets caused the formation of the 5S,12S-DiHETE. It is concluded that the 5S,12S-DiHETE is not derived from leukotriene A4 but is a product of the successive reactions of arachidonic acid with two lipoxygenases of different positional specificities. PMID- 6289380 TI - Effects of prostaglandins on cyclic nucleotide phosphodiesterase activity in the human term placenta. AB - Slices of human full-term placentas, obtained by elective cesarean section, were incubated in the absence or presence of prostaglandins (PGs) and the cyclic AMP phosphodiesterase (cAMP PDE) activity was measured. PGE1 and PGI2 were shown to stimulate cAMP PDE activity. The effect of PGE1 is related to an increase in the Vmax of the low Km activity without alteration of this apparent Km. Several findings suggest that the cAMP PDE is activated by its own substrate; PGE1 and PGI2, promote an increase of cAMP formation which is observed before the cAMP PDE activation. Dibutyryl cAMP or theophylline also activate cAMP PDE. In contrast, PGF2 alpha does not influence either adenylate cyclase or AMP PDE. In addition, we found that the ability of the placenta to degrade cAMP, increases after parturition. PG levels are higher in the foeto-placental unit during labor, and a causal relationship between these two phenomena is possible. Our data supporting the concept of hormonal control of cAMP PDE is consistent with the hypothesis that an accelerated cAMP metabolism in placenta contributes to the maintenance of a constant equilibrium of the cyclic nucleotide levels in the foeto-placental unit. PMID- 6289382 TI - [Perspectives of the clinical genetics development]. PMID- 6289383 TI - Arginine vasopressin deficient Brattleboro rats fail to develop tolerance to the hypothermic effects of ethanol. AB - We have tested the hypothesis that animals with reduced levels of arginine vasopressin (AVP) would show reduced tolerance to ethanol. Brattleboro rats either heterozygous or homozygous for the diabetes insipidus (DI) trait and normal Sprague-Dawley rats were exposed to ethanol vapor for 21 days. Two days later, tolerance was evaluated by monitoring body temperature reductions after intraperitoneal injection of 2 g/kg (20% w/v) ethanol. Under the same conditions of chronic ethanol exposure, Sprague-Dawley rats, but not Brattleboro rats, displayed tolerance to the hypothermic effects of intraperitoneal ethanol. This phenomenon did not appear to be related to differences in ethanol metabolism or blood alcohol levels in Brattleboro rats. These data support a possible role for AVP in the development or maintenance of tolerance. PMID- 6289384 TI - Comparison of the biologic actions of corticotropin-releasing factor and sauvagine. AB - Corticotropin-releasing factor (CRF), a peptide isolated from ovine hypothalamus, and sauvagine, a peptide isolated from frog skin, share significant structural homology and elicit a number of similar biological responses. CRF is more potent than sauvagine in stimulating pituitary ACTH secretion. Sauvagine, however, is 5 10 times more potent than CRF to act within the brain to increase plasma levels of catecholamines and glucose and to elevate mean arterial pressure. Sauvagine is likewise more potent than CRF to act outside the brain to increase superior mesenteric artery flow and plasma glucose concentrations and to decrease mean arterial pressure. CRF and sauvagine produce important effects representative of biologically active peptides. PMID- 6289386 TI - beta-Endorphin release by angiotensin II: studies on the mechanism of action. AB - Blood-borne angiotensin II induces release of beta-endorphin-like immunoreactivity (beta-EI) from rat anterior pituitary gland. To study the mechanism of action we investigated in rats the effect of transection of subfornical organ efferent projections on angiotensin-induced beta-EI release in vivo and also the direct action of angiotensin II on beta-EI release from isolated adenohypophyses in vitro. (i) No effect of transection of subfornical organ efferents on the increase in plasma beta-EI following intravenous infusions of angiotensin II was found. (ii) When anterior pituitary quarters were continuously superfused in vitro, angiotensin II (1-10 nM) caused release of beta EI into the superfusion medium in a dose-dependent manner. The stimulatory effect of angiotensin II (3 nM) was blocked by the receptor antagonist saralasin (300 nM). We conclude that beta-endorphin release by blood-borne angiotensin II, in contrast to other central effects of angiotensin, is not mediated by the subfornical organ; instead a direct action of angiotensin II on the adenohypophysis could be a mechanism of action responsible. PMID- 6289385 TI - Bombesin production by human small cell carcinoma of the lung. AB - A series of continuous cell lines of human small cell carcinoma of the lung (SCCL) have been evaluated for the production of bombesin (BN). In early established cultures BN was detected in the medium of 9 out of 11 cell lines and in 6 out of 7 cell homogenates examined. Levels in the medium were frequently higher in cultures of later passages compared to earlier passages of the same line and low levels developed in the two previously negative cell lines. Plasma concentrations were greater than 80 pmol/l in 2 out of 27 (7%) randomly selected patients with SCCL. A culture (DMS 406) established from the tumor of a patient with the highest plasma level (1240 pmol/l) was the highest producer in vitro. The results indicate that BN, which has been demonstrated immunocytochemically to be present in normal bronchial mucosal cells, is frequently produced by SCCL in vitro but elevated plasma levels are infrequently found in patients with this neoplasm. PMID- 6289387 TI - [Late results in 170 patients operated on for breast cancer]. AB - The authors investigated the late results, or survival at 5 years after the intervention, in 170 patients operated for breast cancer. The total number of survivors at 5 years was of 71 patients, of which 78 belonged to stages I and II, and only 3 to stage III. The authors present their personal attitude for such cases, and stress some of the difficulties encountered in the application of complex therapy. Of the late complications noted, which totalled 58 cases, the authors mention generalized carcinomatosis in 17 patients, pleuro-pulmonary metastases in 14 patients and 11 cases of chronic oedema of the arm, or the so called "thick arm after Halstead operation". PMID- 6289388 TI - [Epidemiological study of hepatitis A using the measurement of anti-VHA antibody]. PMID- 6289389 TI - [Use of the simple radial hemolysis technic for the detection of antibodies to various flaviviruses]. PMID- 6289390 TI - [Analysis of the contraction induced by isoproterenol in isolated cerebral and femoral arteries of cat]. AB - The vasoconstrictor responses induced by isoproterenol in cylindrical segments of posterior communicating and femoral arteries of the cat were analyzed. For this purpose, dose-response curves for isoproterenol before and after addition of propranolol (5 X 10(-9) to 10(-6) M) or phentolamine (10(-6) M) to the bath were determined. Propranolol did not significantly change the contraction induced by isoproterenol in cerebral and femoral arteries. Phentolamine reduced the contractile responses of cerebral arteries evoked by all concentrations of isoproterenol. However phentolamine shifted the dose-response curve to isoproterenol in femoral arteries to the right, the pA2 value for this antagonist being 6.74. These results indicate that the alfa adrenoceptors are involved in the contraction caused by isoproterenol in femoral arteries, whereas in brain arteries other receptors or mechanisms are also likely involved. PMID- 6289392 TI - Structure, expression, and evolution of growth hormone genes. PMID- 6289391 TI - Receptors reconsidered: a 20-year perspective. PMID- 6289393 TI - Glucocorticoid regulation of proopiomelanocortin gene expression in rodent pituitary. PMID- 6289394 TI - The role of nonsteroidal regulators in control of oocyte and follicular maturation. PMID- 6289396 TI - Characteristics of the guanine nucleotide-binding regulatory component of adenylate cyclase. PMID- 6289395 TI - Organization of the thyroid hormone receptor in chromatin. PMID- 6289397 TI - Increased hepatotoxicity of carbon tetrachloride by the hypolipidemic drug nafenopin in rats. AB - Pretreatment of rats with the potent hypolipidemic drug nafenopin enhanced the hepatotoxicity of carbon tetrachloride as judged by the serum activity of glutamic pyruvic transaminase (SGPT), hepatic triglyceride content and glucose-6 phosphatase activity as well as by light and electronmicroscopy. Nafenopin alone caused an increase in liver weight, proliferation of peroxisomes and of smooth endoplasmic reticulum in hepatocytes. No marked changes of serum SGPT activity were observed, but hepatic glucose-6-phosphatase activity decreased. Results suggest that the hypolipidemic compound can act on endoplasmic reticulum membranes so that they become more susceptible to the attack by carbon tetrachloride or it(s) metabolite(s). PMID- 6289398 TI - Effects of chronic histamine and ovalbumin aerosols on pulmonary beta adrenergic receptors in sensitized guinea pigs. AB - Using (3H) dihydroalprenolol, we demonstrated that the concentration of pulmonary beta adrenergic receptor binding sites was lower in guinea pigs showing a marked bronchoconstrictive response to histamine and in ovalbumin-treated guinea pigs. This result suggests that the changed concentration of the pulmonary beta receptors in an animal model of asthma could be resulted from bronchoconstriction stress. PMID- 6289399 TI - Effects of cadmium on the level of serum corticosterone and adrenocortical function in male and female rats. AB - The present study was designated to obtain information about the effects of cadmium (Cd) on the level of serum corticosterone and on the adrenocortical function in male and female rats. The animals were injected twice a day with cadmium chloride at a dose of 1 mg/Kg of body weight for 2 and 7 consecutive days. After Cd treatment for 2 and 7 consecutive days, the adrenal weight increased and the level of serum corticosterone decreased in male and female rats. The steroid response to injected ACTH of the treated rats of both sexes was similar to that of the control rats but there was a marked increase in the disappearance rate of corticosterone from plasma in Cd-treated rats of both sexes. These results suggest that the low level of serum corticosterone in Cd treated rats of both sexes is due to the marked increase in the disappearance rate of corticosterone from plasma. PMID- 6289401 TI - Medullary axonal projections of respiratory neurons of pontile pneumotaxic center. AB - In decerebrate, cerebellectomized, vagotomized, paralyzed and ventilated cats, activities were recorded from the phrenic nerve and single respiratory neurons in the area of the nucleus parabrachialis medialis and Kolliker-Fuse nucleus. Stimuli were delivered in the medulla and cervical spinal cord to elicit antidromic action potentials for these neurons and, hence, establish their axonal projections. Antidromic activation was obtained for 18 of 193 neurons following medullary stimulations. Following spinal stimulations, only two respiratory neurons exhibited some responses characteristic of antidromic activation. In the same pontile areas, a number of neurons with no respiratory-modulated or spontaneous activities were antidromically activated by medullary or spinal stimulations. Results are considered in the context of neuroanatomical studies which have established possible interconnections within the brainstem respiratory control system, and hypotheses for functions of the pontile pneumotaxic center in ventilatory control. PMID- 6289400 TI - The effect of partial ileal bypass on receptor-mediated uptake and catabolism of low density lipoprotein in the rhesus monkey. AB - The fractional rates of catabolism (FCR) of 125I-labelled low-density lipoprotein (LDL) and 131I-labelled LDL coupled with 1,2-cyclohexanedione (LDL-CHD) were determined before and 6 weeks after partial ileal bypass in three Rhesus monkeys. Receptor-mediated catabolism, determined as the difference between the FCRs of 125I-LDL and 131I-LDL-CHD, increased significantly after partial ileal bypass. Analysis of tissue distribution of radioactivity in one monkey, killed at the conclusion of a third turnover study 6 months after partial ileal bypass, suggested that the liver was the main tissue site of accumulation of LDL, and that uptake had occurred mainly via a receptor-mediated mechanism. It is concluded that partial ileal bypass stimulates receptor-mediated LDL catabolism in the Rhesus monkey as in man, and that this increase is probably mediated via hepatic LDL receptors. PMID- 6289403 TI - [Primary liver carcinoma. Clinicopathological study of 109 patients []. PMID- 6289402 TI - [High fiber diet prepared with regional foods as an aid in the control of patients with diabetes []. PMID- 6289404 TI - Schistosomiasis: nutritional implications. AB - Most studies on the interactions of nutrition and schistosomiasis have involved experimental animals. Generally, these studies show that schistosomiasis may be alleviated by severe malnutrition, especially calorie deprivation. However, nutritional modulation of schistosomiasis in experimental animals is observed only when severe deficiencies are induced during the acute disease. Except during overt famine, in humans such severe forms of malnutrition occur mainly in the young, who are not yet exposed to heavy burdens of schistosomal infection. Older individuals, who may be heavily infected, are usually not so severely malnourished. Therefore, the nutritional rehabilitation of severely malnourished children, which should improve immune function, is unlikely to exacerbate the immune-mediated pathogenesis of schistosomal disease. Nutritional interventions can be planned independently of the prevalence or intensity of schistosomiasis in the population. Heavy infection with Schistosoma mansoni can cause both direct nutritional losses and severe disease manifestations. These constitute two important reasons for the administration of mass chemotherapy to the heavily infected segment of a population. Both of these adverse effects can be corrected by a reduction in the intensity of infection; complete cure is not necessary. PMID- 6289405 TI - Amebiasis: nutritional implications. AB - Studies on the role of nutrition in amebiasis in humans and experimental animals are meager. Some reports suggest that malnutrition of the host increases the incidence of infection and potentiates the severity of the disease. Others suggest that malnutrition protects the host against invasion. A few reports indicate that dietary regimens can alleviate symptoms and even eradicate the parasite. Others doubt a correlation between diet and rate of infection or disease manifestations. The problem is complex because the ameba is influenced by its own diet, which in turn depends on the host's diet, the bacterial flora of the gut, and coexisting infections. The host is variously altered by dietary depletions and supplementations, which affect susceptibility and resistance, and by the presence of other disease conditions. Carefully designed and executed studies of infections in humans and experimental animals, combined with studies in vitro of the nutritional requirements and physiology of the parasite, are needed for definition of the influence of host nutrition in amebiasis. PMID- 6289406 TI - Receptor sites for mycoplasmal viruses on Acholeplasma laidlawii. AB - All three groups of mycoplasmal viruses tested, MVL1, MVL2, and MVL3, were inactivated by both membranes and lipoglycan from Acholeplasma laidlawii strain JA1. The interaction was specific for components of A. laidlawii since no inactivation occurred with either membrane or lipoglycan from Acholeplasma oculi. Adsorption of virus was proved by the demonstration of radiolabeled virus lipoglycan or virus-membrane particles that were separable in density gradients from either component alone. Receptors essential for adsorption of MVL2 are part of the oligosaccharide chain of the lipoglycan since deacylation not only did not interfere but actually increased adsorption of virus per unit weight. More complex interactions involving membrane proteins were observed for MVL1 and MVL3. PMID- 6289407 TI - Guanine-plus-cytosine content, hybridization percentages, and EcoRI restriction enzyme profiles of spiroplasmal DNA. AB - The guanine-plus-cytosine (G + C) content of spiroplasmal DNA was calculated from the melting temperature determined spectrophotometrically and the buoyant density determined by equilibrium density gradient centrifugation in CsCl. Only two ranges of G + C values were found: 25-27 mol% and 29-32 mol%. The DNA of the following spiroplasmas has 25-27 mol% G + C: Spiroplasma citri (serogroup I-1); the spiroplasmas pathogenic to the honeybee (KC3, BC3, and B63; serogroup I-2); the corn stunt strain (E275; serogroup I-3); the tick strain 277F (serogroup I 4); the drosophila strain (serogroup II); and one group of flower spiroplasmas (serogroup III). The DNA of a second group of flower spiroplasmas (serogroup IV) and the SMCA strain (serogroup V) has a G + C content of 29-31 mol/. The classification of flower spiroplasmas into two groups on the basis of G + C content agrees well with the groupings based on serologic and protein analysis. Spiroplasmas isolated from honeybees in Morocco (B13) or froghoppers in Corsica (L89) have 29-31 mol% G + C, a value that corroborates the relatedness of these strains and the flower spiroplasmas of serogroup IV found by serologic analysis. Reannealing experiments between the vivo-labeled DNA of S. citri and unlabeled DNA of other spiroplasmas gave the following percentages of hybridization: 64% with honeybee spiroplasma DNA, 49% with corn stunt spiroplasma DNA, and 19% with tick spiroplasma 277F DNA; no significant hybridization was observed with DNA of any other spiroplasma. The taxonomic position of the tick spiroplasma 277F within serogroup I was confirmed by hybridization experiments involving [3H]DNA of this strain. The value of polyacrylamide gel analysis of DNA fragments produced by the action of EcoRI restriction enzyme on DNAs from various spiroplasmas is also discussed. PMID- 6289410 TI - [Giant Abrikosov's tumor located in the abdominal wall. Surgical treatment]. PMID- 6289409 TI - Molecular organization and selective solubilization of lipids and proteins in the envelope of mycoplasmal virus L2. AB - The composition and molecular organization of mycoplasmal virus L2 (MVL2) were studied and compared with those of the cell membrane of the host, Acholeplasma laidlawii strain JA1. The virus contained 0.2-0.25 mumol of polar lipids per mg of viral protein. The lipid species of the MVL2 were the same as those of the host cells. Nevertheless, the proportions between the various polar lipids were different, with a much lower content of phosphatidylglycerol in the viral lipids. Despite these quantitative differences, the fatty acid composition of MVL2 was similar to that found in the host cells, a similarly allowing alteration of the fatty acid composition and study of its effect on viral absorption, penetration, and release. Pulse-chase experiments revealed that the lipids incorporated into the virus were synthesized before and after infection. Electron paramagnetic resonance spectrometry suggested that the viral lipid domain had the properties of a lipid bilayer. Nevertheless, the hydrocarbon chains in the MVL2 envelope were less mobile than those in membranes of the host cells, a difference apparently due to the different content, composition, and disposition of proteins in the MVL2 envelope. The electrophoretic pattern of MVL2 polypeptides was dominated by four major and five minor bands distinct from the polypeptides present in A. laidlawii membranes. None of the polypeptides gave a positive periodic acid-Schiff reaction, a result suggesting the absence of glycoproteins. Selective solubilization experiments excluded the possibility that one or more of the major polypeptides was associated with a capsid structure. Lactoperoxidase mediated iodination of intact viral particles revealed that at least two of the major polypeptides were localized on the external surface of the viral envelope. The susceptibility of these polypeptides to brief proteolytic treatment and the finding that the infectivity of the virus was dramatically affected by such treatment suggested that these polypeptides were playing a role in recognition and/or attachment of the virus to the host cells. PMID- 6289408 TI - Cholesterol and phospholipid uptake by mycoplasmas. AB - Mycoplasma offer several unique advantages for investigating the mechanism controlling transfer and uptake of exogenous cholesterol and phospholipids by biomembranes, as their plasma membrane interacts directly with exogenous lipid donors and their endogenous lipid synthesis is restricted. Growing cells of five species of Mycoplasma were found to take up significant quantities of phosphatidylcholine and sphingomyelin as well as free and esterified cholesterol. In contrast, growing cells of three species of Acholeplasma failed to take up any of the exogenous phospholipids and incorporated only low amounts of free cholesterol and no esterified cholesterol. It is hypothesized that Mycoplasma species have receptors for serum lipoproteins and phospholipid-cholesterol vesicles that facilitate the transfer of cholesterol and phospholipids to the growing cell membrane. Our finding that gentle trypsin treatment of growing Mycoplasma capricolum cells decreased their cholesterol uptake ability by about 50% but did not affect cholesterol uptake by growing Acholeplasma laidlawii cells appears to support the existence of protein receptors for lipoproteins on the surface of Mycoplasma but not on Acholeplasma species. Digestion of membrane phospholipids by phospholipase A2 decreased the cholesterol-binding capacity of isolated A. laidlawii and M. capricolum membranes, roughly in proportion to the amount of phospholipid digested. The total removal of phosphatidylglycerol and diphosphatidylglycerol from A. laidlawii membranes by phospholipase A2 only decreased but did not abolish cholesterol uptake, an indication that glycolipids also participate in cholesterol uptake. PMID- 6289411 TI - [Centenary of the discovery of the Koch bacillus]. PMID- 6289412 TI - [100 years after the discovery by Robert Koch of the etiological agent of tuberculosis]. PMID- 6289413 TI - [Quantitative relations in bacteriological research in tuberculosis]. AB - The authors have carried out a comparative analysis of the sensitivity of the microscopic examination as compared with that of the bacteriologic cultures in the evidencing of the Koch bacilli in biologic products obtained from new cases of tuberculosis, before the start of any kind of treatment with tuberculostatics. The authors attempted to establish the proportion of positive results with the two tests in the conditions of an increased number of slides to be examined by microscopic methods from 1 to 4. At the same time other reciprocal quantitative relationships have been determined between these two tests, and differences were evidenced at the same time between the positive results obtained in symptomatic and asymptomatic patients. It was concluded that for achieving an optimal output with bacteriological investigations of tuberculosis the ideal solution is to examine 3 slides prepared from a specimen, and to seed 3 culture tubes, or 4 tubes for asymptomatic patients. PMID- 6289414 TI - [Bacteriological and epidemiological observations on atypical mycobacteria isolated at the laboratory of the Cluj-Napoca Tuberculosis Hospital]. AB - Excluding cases of clinically manifest mycobacteriosis atypical mycobacteria can be detected in the excreta of healthy subjects, of tuberculous patients, or of other patients suffering from other diseases, without any major clinical implication. In certain environment conditions however epidemiologic manifestations may be induced by these organisms, of the hospitalism type. Atypical strains of the mycobacteria isolated over a period of 10 years represent 3.17% of all isolated strains. The highest percentage according to the Runyon groups were found in the IV-th group (43%), followed by the II-nd group (with 42%), and the III-rd group (with 11%). Diffusion of mycobacteria from the II-nd and the IV-th Runyon groups has determined hospitalism phenomena that developed in two distinct epidemic outbreaks determined by different factors. PMID- 6289415 TI - [Aspects of dysmicrobism in the isolation of Koch bacilli and its pathological implications]. AB - The results are presented, of a follow-up over several years concerning the changes which occurred in the products sampled from tuberculous patients, with regard to the ecologic balance between Mycobacterium tuberculosis, and other mycobacteria and fungi under the influences of chemical drugs and antibiotics. Thus atypical mycobacteria have been recorded, in a proportion of 1.07% (and in some periods as high as 2.52%). Other species were also identified, classified according to morphological and chemical characteristics. These species are included in a synoptic scheme used in laboratories which are concerned with the isolation of mycobacteria. At present different species of mycobacteria can be isolated on the same culture medium, as appears also from the results of the authors of this study. Beside manifestations of dysmicrobism another fact was also demonstrated, namely the frequent isolation of fungi in patients undergoing anti-tuberculous therapy, or secondary to the anti-tuberculous treatment. This has permitted, on the basis of the authors' cases to confirm some pulmonary mycoses according to generally accepted rigorous laboratory techniques. The present study stresses the new pathologic phenomena interrelated with tuberculosis. These data expand the knowledge of laboratory techniques and methods, and improve cooperation between laboratories and clinical units involved in the diagnosis and treatment of pulmonary affections. PMID- 6289416 TI - [Arguments for extending morphological research in pneumological practice]. PMID- 6289417 TI - [Reminiscences of the scientific work of Robert Koch]. PMID- 6289418 TI - [Economic and social implications of post-tuberculosis syndromes]. AB - The fact that 30.5% of patients with post-tuberculous syndromes belonging to a category of active population have been pensioned off for health reasons is a demonstration that these syndromes may play a significant role in determining the incapacity for work. The practical conclusion is that the prophylaxis of post tuberculous syndromes--early diagnosis, chemotherapy with new drugs, and kinesitherapy--in all cases of tuberculosis at their first treatment may determine a deceased incidence of these syndromes. The reduction of the percentage of post-tuberculous syndromes recorded for 100 cases of tuberculosis treated with modern therapeutic regimens will result in a collective benefit consisting in a lower pressure on the general and specialized sanitary network with regard to the number of procedures provided. The health network potentialities could this be used for the solving of other problems of public health. At the same time by reducing the percentage of post-tuberculous syndromes, important funds can be spared, which are used for treatment, sick leave pay, and invalidity pensions. The continuous improvement of the organizational frame of health protection in our country should consider also the official regulatory of the medico-social status of patients with post-tuberculous syndromes. PMID- 6289420 TI - [Case of poisoning as a result of ingesting fish containing ciguatoxin]. PMID- 6289419 TI - Ketoconazole in paracoccidioidomycosis. A long-term therapy study with prolonged follow-up. PMID- 6289421 TI - [Meningoencephalitis associated with orbivirus infection]. PMID- 6289422 TI - Preparation of ferritin conjugates and antibodies for the localization and identification of antigens in tissues and cells by electron microscopy. AB - Ferritin, due to its high electron density, serves s a good marker in immunoelectron microscopic studies to localize antigens on and in cells and tissues. A step-by-step methodology for the conjugation of antibodies to ferritin is presented. Special attention has been placed on the techniques for the purification of ferritin and antibodies, conjugation procedures, and the treatment of specimens with conjugates. Methods for obtaining the separation of conjugated antibodies from unconjugated antibodies and for testing the purity and specificity of the labeling reagents are discussed. PMID- 6289423 TI - The effects of ionizing irradiation on the ciliated cells of the central nervous system (CNS) in man--a scanning electron microscopy study. AB - In order to investigate the effects of radiotherapy on the ciliated ependymal lining of the CNS, samples were taken at autopsy from the lateral ventricles down to the IVth ventricles in patients irradiated for Glioblastoma multiforme in one hemisphere. Doses higher than 20 Gy affected the cilia and the lesions were more pronounced the heavier the dose. No regeneration of cilia were found in those regions which had received more than 20 Gy. In the areas irradiated with 20 - 15 Gy the cilia had a more normal form and cilia irradiated with less than 15 Gy were considered to be normal. PMID- 6289424 TI - Characterization of marrow-derived adherent cells. Evidence against an endothelial subpopulation. AB - Cultured, marrow-derived, adherent cells (MDAC) provide a microenvironment which supports the proliferation of haemopoietic stem cells (HSC) for extended periods in vitro. Morphological characterization suggested that MDAC populations consisted of a variety of cell types, including mononuclear phagocytes, fibroblastoid cells, fat cells and vascular endothelial cells. Recently performed functional characterization studies suggest that they consist largely of collagen producing, fibroblastic cells. MDAC were not, however, examined systematically for endothelial cell characteristics. Unrecharged cultures of MDAC, shown in parallel studies to support in vitro haemopoiesis, were examined for endothelial cell markers. These included the presence of Weibel-Palade bodies and synthesis of factor VII related antigen. They were also examined biochemically for synthesis of basement membrane (type IV) collagen. The results of these investigations were negative in all cultures examined. It is thus concluded that vascular endothelial cells are not present as a significant component of the unrecharged MDAC population and do not, therefore, contribute to the functional haemopoietic microenvironment in vitro or in vitro. PMID- 6289425 TI - Ischemic tissue injury--peripheral nerves. PMID- 6289426 TI - Distribution of cyclic AMP in human seminal plasma and its relation to sperm progressive motility. AB - Organelles and amorphous substance (pellet II) isolated from human seminal plasma contained 3'5' AMP (cyclic AMP, cAMP) in manifold smaller amounts than did the particle-free seminal plasma. The amount of cAMP associated with pellet II did not differ significantly between normospermic and oligozoospermic or teratozoospermic ejaculates. In analyses of split ejaculate fractions, the distribution of cAMP coincided with that of fructose and protein (but not with the Mg2+- and Ca2+-dependent ATPase activity or with zinc), indicating secretion of cAMP by the seminal vesicles. The distribution profiles of cAMP in the various ejaculate fractions were similar for particle-free seminal plasma and for pellet II material. The cAMP contents of the fractions were compared with sperm motility in the same fractions. An inverse relationship was found, with the first three fractions displaying higher sperm motility and the last three fractions higher cAMP content. PMID- 6289427 TI - [Fiasco in medicine?]. PMID- 6289428 TI - [Signet ring cell carcinoma of the small intestine in sheep]. PMID- 6289429 TI - [Composite recipe No. II for coronary heart disease and platelet functions]. PMID- 6289430 TI - [The performing-mechanism for hormone of phosphatidylinositol system]. PMID- 6289431 TI - Cell biology yields clues to lung cancer. PMID- 6289432 TI - Are ions involved in the gating of calcium channels? AB - The rates of activation and deactivation of the currents carried by calcium, strontium, or barium ions through the voltage-sensitive calcium channel of Paramecium are different. The differences cannot be attributed to complications due to internal ion concentration, calcium channel inactivation, potassium current activation, surface charge effects, or incomplete space clamping. The findings indicate participation of the divalent cations in the voltage-driven calcium channel gating process. PMID- 6289433 TI - Postural asymmetry and movement disorder after unilateral microinjection of adrenocorticotropin 1-24 in rat brainstem. AB - A unilateral microinjection of adrenocorticotropin 1-24 in the rat brainstem in the region of the locus ceruleus resulted in postural asymmetry and movement disorder that resembled human dystonia, the severity and duration (2 to 3 days) being dose-dependent. These results show for the first time that neuropeptides in the brainstem may modulate posture and movement, and they suggest that some forms of movement disorder such as dystonia may be due to a disordered regulation of postural and locomotor mechanisms by adrenocorticotropin 1-24. PMID- 6289434 TI - An estrogen-binding protein and endogenous ligand in Saccharomyces cerevisiae: possible hormone receptor system. AB - A protein macromolecule in the cytosol of the unicellular eukaryotic yeast Saccharomyces cerevisiae selectively binds the vertebrate estrogen hormone 17 beta-estradiol with high affinity. Lipid extracts of the yeast cells or the conditioned growth medium yield a substance that can bind competitively to the tritiated estradiol-binding sites in the yeast and to mammalian estrogen receptors. These findings suggest that the binding protein may be a primitive hormone receptor and that the lipid-extractable substance represents the endogenous ligand. PMID- 6289435 TI - Transposition of cloned P elements into Drosophila germ line chromosomes. AB - Recombinant DNA carrying the 3-kilobase transposable element was injected into Drosophila embryos of a strain that lacked such elements. Under optimum conditions, half of the surviving embryos showed evidence of P element-induced mutations in a fraction of their progeny. Direct analysis of the DNA of strains derived from such flies showed them to contain from one to five intact 3-kilobase P elements located at a wide variety of chromosomal sites. DNA sequences located outside the P element on the injected DNA were not transferred. Thus P elements can efficiently and selectively transpose from extrachromosomal DNA to the DNA of germ line chromosomes in Drosophila embryos. These observations provide the basis for efficient DNA-mediated gene transfer in Drosophila. PMID- 6289436 TI - Genetic transformation of Drosophila with transposable element vectors. AB - Exogenous DNA sequences were introduced into the Drosophila germ line. A rosy transposon (ry1), constructed by inserting a chromosomal DNA fragment containing the wild-type rosy gene into a P transposable element, transformed germ line cells in 20 to 50 percent of the injected rosy mutant embryos. Transformants contained one or two copies of chromosomally integrated, intact ry1 that were stably inherited in subsequent generations. These transformed flies had wild-type eye color indicating that the visible genetic defect in the host strain could be fully and permanently corrected by the transferred gene. To demonstrate the generality of this approach, a DNA segment that does not confer a recognizable phenotype on recipients was also transferred into germ line chromosomes. PMID- 6289437 TI - Gene transfer into the Drosophila germ line. PMID- 6289439 TI - Inhibition of adrenocorticotropic hormone secretion in the rat by immunoneutralization of corticotropin-releasing factor. AB - Intravenous administration of rabbit antiserum to ovine corticotropin-releasing factor (CRF) markedly reduced the CRF-induced rise of plasma adrenocorticotropic hormone (ACTH) in intact nonstressed adult male rats while blocking more than 75 percent of the ACTH release observed in rats exposed to ether stress. Furthermore, antiserum to CRF significantly lowered ACTH levels in adrenalectomized animals. These results suggest that endogenous CRF plays a physiological role in regulating ACTH secretion. PMID- 6289438 TI - Hormonal regulation of gonadotropin receptors and steroidogenesis in cultured fetal rat tests. AB - Gonadotropic activation of the adult rat testis in vitro and in vivo is followed by down-regulation of luteinizing hormone receptors and decreased androgen responses to subsequent hormonal stimulation. In contrast, treatment of cultured fetal testes with gonadotropins and dibutyryl adenosine 3',5'-monophosphate enhanced steroidogenic responsiveness and did not cause the luteinizing hormone receptor loss and desensitization that is characteristic of the adult gonad. The analysis of gonadotropin receptors and action in cultured fetal testis cells facilitates developmental studies of gonadal function, and has revealed significant differences in the responses of fetal and adult Leydig cells to gonadotropic regulation. PMID- 6289440 TI - Herpes simplex virus type-1 glycoprotein D gene: nucleotide sequence and expression in Escherichia coli. AB - The protein coding region of the herpes simplex virus type-1 glycoprotein D (gD) gene was mapped, and the nucleotide sequence was determined. The predicted amino acid sequence of the gD polypeptide was found to contain a number of features in common with other virus glycoproteins. Insertion of this protein coding region into a bacterial expressor plasmid enabled synthesis in Escherichia coli of an immunoreactive gD-related polypeptide. The potential of this system for preparation of a type-common herpes simplex virus vaccine is discussed. PMID- 6289441 TI - Opiate receptor distribution in the cerebral cortex of the Rhesus monkey. AB - The distribution of opiate receptors in the cerebral cortex of the rhesus monkey (Macaca mulatta) was determined by autoradiographic visualization of [3H]naloxone binding to tissue sections. Naloxone was bound in relatively large amounts to the cortical laminae containing the cell bodies of output neurons, to a varying set of additional laminae in different cortical fields, to fields closer to more primitive types of cortex, and to polysensory cortical fields. From these laminar and areal variations in distribution, it appears that opiate receptors play a role in specific aspects of cortical function. PMID- 6289442 TI - Molecular biology of learning: modulation of transmitter release. AB - Until recently, it has been impossible to approach learning with the techniques of cell biology. During the past several years, elementary forms of learning have been analyzed in higher invertebrates. Their nervous systems allow the experimental study of behavioral, neurophysiological, morphological, biochemical, and genetic components of the functional (plastic) changes underlying learning. In this review, we focus primarily on short-term sensitization of the gill and siphon reflex in the marine mollusk, Aplysia californica. Analyses of this form of learning provide direct evidence that protein phosphorylation dependent on cyclic adenosine monophosphate can modulate synaptic action. These studies also suggest how the molecular mechanisms for this short-term form of synaptic plasticity can be extended to explain both long-term memory and classical conditioning. PMID- 6289443 TI - Receptors for maleylated proteins regulate secretion of neutral proteases by murine macrophages. AB - Receptors for maleylated or acetylated proteins as well as for alpha-2 macroglobulin-protease complexes on macrophages serve as scavengers by mediating the uptake of macromolecules from the extracellular compartment. Described in this report is a novel function of these receptors on macrophages: regulation of neutral protease secretion. The binding of maleylated bovine serum albumin to macrophages triggered secretion of three neutral proteases: neutral caseinases, plasminogen activator, and cytolytic proteinase. Release of acid phosphatase, however, was not induced. An important biological consequence of protease secretion by macrophages, tumor-cytolysis, was also triggered by engagement of the receptor for maleylated bovine serum albumin. By contrast, the binding of alpha-2-macroglobulin-protease complexes to the macrophages suppressed secretion of all three proteases. Thus two receptors heretofore believed to serve principally as scavengers also regulate secretory functions of macrophages. PMID- 6289444 TI - [Value of the study of the kinetics of 85 strontium for the classification of osteogenic sarcomas ]. AB - Because of the polymorphism of osteogenic sarcomas, examination of a biopsy specimen, which is necessarily small, cannot give an accurate evaluation of the differentiation of the whole tumor. Therefore, a test which provides an overall assessment of the functional capacity of the tumor is needed. For the last ten years, we have been using 85 strontium, which has a metabolism similar to that of calcium and radioactive characteristics that allow external measurements. With this isotope, classification of osteogenic sarcomas is more accurate, ensuring better therapeutic trials. PMID- 6289446 TI - [Bilateral lung irradiation as adjuvant therapy in osteogenic sarcoma ]. AB - Bilateral lung irradiation as well as chemotherapy are still controversial in the "prevention" of pulmonary metastases from osteosarcomas. This paper presents theoretical and experimental evidence in support of such an irradiation. Doses which can be tolerated by the normal lung are recalled. Data from clinical experience is analyzed and the optimal modalities of irradiation are described. Confirmation of the presumptive effectiveness of radiation therapy can be obtained only through controlled trials such as the one which is ongoing at the EORTC. PMID- 6289445 TI - [Osteogenic sarcomas of the limbs. Radical treatment of the primary tumor: choosing between irradiation and amputation ]. AB - In osteogenic sarcomas of the extremities, radical therapy of the primary tumor now seems more precisely delineated. In most cases, especially in large tumors with lytic lesions or fractures, and in children. Irradiation is indicated only in small tumors with condensation, and in tumors of the proximal metaphysis of the humerus. Conservative surgical treatment with carcinologically satisfactory resection of involved bone followed by reconstruction, is currently under evaluation. Such treatment is indicated in specific cases: children in the final stage of growth, and small tumors without major involvement of soft tissues that respond to preoperative chemotherapy. PMID- 6289447 TI - [Adjuvant therapy in the management of osteosarcomas: the O3 trial (EORTC and ISPO) ]. AB - The European Organization for Research on the Treatment of Cancer (EORTC) and the international Society of Pediatric Oncology (ISPO) have set up a randomized controlled trial designed to compare three different regimens of adjuvant therapy applied after treatment of the primary tumor (usually by radical surgery): a prolonged course of chemotherapy (41 weeks) combining adriamycin, methotrexate in high doses followed by folinic acid, vincristine, and cyclophosphamide; radiotherapy delivering 20 grays to the lungs; and a short course of chemotherapy (8 weeks) combining adriamycin, methotrexate in high doses, folinic acid and vincristine, associated with irradiation of the lungs as in the preceding protocol. At the time being, 156 patients have been included in this study. If conclusions are to be drawn from this trial, a total of 300 patients seems necessary. PMID- 6289448 TI - [Changes in the management of osteosarcomas of long bones in the Leon Berard Center, in the last twenty years ]. AB - From 1958 to 1977, 68 patients with long bone osteosarcoma were treated with high dose radiation (greater than 80 grays) followed by amputation when needed. The survival rate free of disease is 33% at three years (23/68) and 31% at five years (20/64). Sterilization of the limb was obtained in 72% of cases (18/25 amputation specimens). Preservation of a functional limb often proved impossible because of the frequent occurrence of trophic disorders, especially fractures, after radiotherapy. The main benefit of primary irradiation is to avoid amputations that have become useless because pulmonary metastases have arisen. Surgical excision of pulmonary metastases can lead to prolonged survival (3 out of 6 patients alive free of disease at three years). As no conclusive randomized trials are as yet available, the benefits of adjuvant therapy (chemotherapy and/or radiotherapy) are still prospective. The present policy is to use adjuvant therapy in all cases, conservative treatment for small lesions, primary amputation in children under fourteen, and primary irradiation followed by systematic amputation one year later for extensive lesions in older patients. PMID- 6289449 TI - [An experimental model of osteosarcomas in rats ]. AB - Satisfactory experimental models for preclinical prediction in cancerology must answer the following criteria: reproducibility of the method used for inducing tumors; clinical, pathological and kinetic similarity with the corresponding human tumors. We have developed a model of osteosarcoma locally induced by insoluble radioactive cerium chloride (144Ce CI3) in Sprague Dawley rats. This method yields over 80% of bone tumors at the injection site, of which approximately half are histologically similar to human tumors. These tumors double their volume fairly slowly (in approximately 20 days); lung metastases occur both early and frequently (80% of animals). A transplantable tumor was developed from an induced osteosarcoma and adapted to the Curie strain. Transplantation in the bone, next to the bone, or under the skin is followed by widespread metastatic dissemination. The kinetics and histological features of the primary tumor are maintained. Tumor 85 strontium uptake is similar to that seen in human osteosarcomas. These new models of osteosarcomas are being used for evaluating new cancer chemotherapeutic agents and interferon, etc. PMID- 6289450 TI - [Ejaculation problems in traumatic paraplegia ]. AB - Basing themselves on 149 cases, the authors describe the spinal neurophysiology of ejaculation in spinal man, and put forward certain hypotheses. They suggest that the reflex of seminal emission is a lumbar reflex, which may be dependent on conductive factor originating in the upper neurological structures, and that the orthosympathetic system, which leaves the spinal cord through the thoracolumbar junction, could provide a highway for the psychological nerve impulses inducing seminal emission. PMID- 6289451 TI - [Acute pancreatitis. Seventy-one cases ]. AB - This article reviews 71 cases of acute pancreatitis on the basis of 153 criteria. All the patients concerned underwent operations or anatomical checkups. The indications and results of the different types of operation are analysed. It is the authors' policy to operate as soon as possible in order to assess the gravity of the lesions. This attitude is justified by the risk of unsuspected biliary stones and the uncertain evolution of extravasated peripancreatic effusion tracks. Strictly controlled drainage still has its place in the treatment of pancreatitis, alongside of exeresis and necrosectomy. PMID- 6289452 TI - [Cytological complications of IUDs ]. AB - The cytological modifications due to IUDs are incompletely or inaccurately known to clinicians and cytologists. IUDs of all types bring about a modification of the maturation index, and particularly of the estrogenic index which, throughout the cycle, appears to be lower than that of a control group. The presence of a sub-normal inflammatory contest in smears of IUD users is no proof of an increased incidence of microbic infections. Presently there is no sign of any carcinogenic role played by IUDs. The papillary metaplasia induced by IUDs appeared more frequently than squamous endometrial metaplasia. PMID- 6289453 TI - [Peroxisomes and hypolipidemic agents ]. AB - Peroxisomes (microbodies) have been localized in the cytoplasm of various cells, mainly the hepatocytes and the renal tubular cells. Various enzymes were found in the granular matrix and sometimes in the nucleus; they were involved in the production and the degradation of hydrogen peroxide, in regulation of the energetic processes of the cell, and in the oxidation of fatty acids. Several xenobiotica and drugs such as hypolipidemic agents clofibrate-like, have induced in rodents a proliferation of peroxisomes and frequently, the appearance of malignant tumors. In men, the precise role of peroxisomes in the physiology of the cell, their link with the hypolipidemic action of some drugs and the development of cancer remain to be established. PMID- 6289454 TI - Renal radiopharmaceuticals--an update. AB - Noninvasive radionuclide procedures in the evaluation of renal disease have been accepted increasingly as effective and valuable alternatives to older clinical methods. The development of suitable radiopharmaceuticals labeled with high photon intensity radionuclides and with 99mTc in particular has stimulated this modality during the last few years. Currently several nearly ideal agents are available for anatomical and functional studies of kidney imparting very low absorbed radiation doses. These include 99mTc-GHA and 99mTc-DMSA for renal morphology and differential function evaluation, 99mTc-DTPA for GFR and 123I orthoiodohippurate for ERPF measurements. A suitable agent as a replacement for the latter labeled with 99mTc is actively being sought. Computer-assisted processing of dynamic renal function studies enables the observer to obtain a wealth of information related to the renal extraction, uptake, parenchymal transit and pelvic transit parameters of the agent administered into the bloodstream. Each of these parameters either globally or differentially contributes to a detailed evaluation of renal disease states. Several of these procedures have been validated against classical techniques clinically but more detailed information is being sought with the recently introduced radiopharmaceuticals. With the detailed validation and increasing recognition of the clinical utility of several of the radionuclidic procedures at many centers, it is hoped that radionuclide assessment of renal disorders ultimately will be made available routinely at all medical facilities. PMID- 6289455 TI - Nuclear renal imaging in acute pyelonephritis. AB - Patients with acute pyelonephritis may present with a spectrum of clinical signs and symptoms. There are few noninvasive diagnostic studies, however, to confirm or exclude this diagnosis. A small number of patients, generally those with severe disease, will demonstrate radiographic changes on excretory urography, but the lack of sensitivity of the IVP in early, acute pyelonephritis is well documented. Several radionuclide techniques have been proposed to assist in the earlier detection of this clinical problem including imaging with Mercury-197 chlormerodrin, Gallium-67 citrate, Technetium-99m glucoheptonate. Technetium-99m DMSA, and, more recently, Indium-111 labeled white blood cells. The success of the renal cortical imaging agents as well as those which localize in infection are described in this report. There appears to be a complimentary role or the cortical imaging agents and the radiopharmaceuticals which localize in bacterial infection. Cortical agents offer the advantage of specific assessment of functioning renal tissue and a convenient, rapid method for following the response to treatment in a noninvasive manner. A pattern is described which may be diagnostic; correlation with Gallium-67 citrate of Indium-111 WBCs may increase the probability of infection as the cause for the cortical abnormality. The measurement of differential renal function using cortical agents provides additional information to assist the clinician in predicting the late effects of infection. Improved sensitivity and specificity, and a reproducible method for following the response to therapy in patients with acute pyelonephritis are the advantages of the techniques described. PMID- 6289456 TI - Radionuclides in the evaluation of urinary obstruction. AB - Radionuclide renography and renal scanning techniques are ideally suited to the initial and follow-up evaluation of patients with obstructive uropathy. While other modalities are superior in their ability to provide anatomic information, the radionuclide study yields functional information for each kidney without the necessity to resort to invasive studies. In addition, the Nuclear Medicine study is well suited to the evaluation of obstruction where serial studies often are required because of a lower radiation burden compared to urography. This lower radiation dose is especially important in obstruction because of the recurrent nature of several kinds of obstructive uropathy and because of the high incidence in the pediatric age group. The ability to control urine flow rate during the procedure through dehydration or administration of diuretics is an additional benefit. Increasing availability of computerization of nuclear medicine procedures as well as interest in studies employing physiologic intervention (including the diuresis renogram) have assured an important place for radionuclide studies in the evaluation of patients with urinary obstruction. PMID- 6289458 TI - Introduction: a success history for surgery and chemotherapy--gestational trophoblastic neoplasia. PMID- 6289457 TI - Nuclear medicine in acute and chronic renal failure. AB - The diagnostic value of renal scintiscans in patients with acute or chronic renal failure has not been emphasized other than for the estimation of renal size. 131I OIH, 67gallium, 99mTcDTPA, glucoheptonate and DMSA all may be valuable in a variety of specific settings. Acute renal failure due to acute tubular necrosis, hepatorenal syndrome, acute interstitial nephritis, cortical necrosis, renal artery embolism, or acute pyelonephritis may be recognized. Data useful in the diagnosis and management of the patient with obstructive or reflux nephropathy may be obtained. Radionuclide studies in patients with chronic renal failure may help make apparent such causes as renal artery stenosis, chronic pyelonephritis or lymphomatous kidney infiltration. Future correlation of scanning results with renal pathology promises to further expand nuclear medicine's utility in the noninvasive diagnosis of renal disease. PMID- 6289459 TI - Immunology of gestational trophoblastic neoplasms. PMID- 6289460 TI - Invasive mole. PMID- 6289461 TI - Beta-subunit hCG and the control of trophoblastic disease. PMID- 6289462 TI - Nonmetastatic and low-risk metastatic gestational trophoblastic neoplasms. PMID- 6289463 TI - The management of poor prognosis trophoblastic disease. PMID- 6289464 TI - The role of surgery in the management of gestational trophoblastic disease. PMID- 6289465 TI - Surgery in metastatic trophoblastic disease. PMID- 6289466 TI - Gestational trophoblastic disease in Hong Kong. PMID- 6289467 TI - Difficult cases of trophoblastic disease and its classification and management. PMID- 6289468 TI - New chemotherapeutic agents in the management of gestational trophoblastic disease. PMID- 6289469 TI - Pneumocystis carinii pneumonia in an apparently nonimmunocompromised patient. PMID- 6289470 TI - Malignant fibrous histiocytoma of the breast. PMID- 6289471 TI - Villonodular synovitis: pigmented and nonpigmented variations. AB - During the past 20 years, 34 cases of pigmented villonodular synovitis have been seen at Jackson Memorial Hospital, University of Miami Medical Center. There were 18 cases in the knee, eight in the hip, one in the elbow, two in the finger, three in the ankle, one in the wrist, and one in the sternoclavicular joint. In reviewing the pathology slides, it was difficult to differentiate fibrous histiocytoma from villonodular synovitis. The cause is considered to be inflammatory. The treatment is subtotal synovial resection with joint replacement as necessary. Recurrences after synovectomy are rare, with none in our series of 34, and only two cases lost to follow-up. I believe synovectomy and debridement to be the treatment of choice, although in the hip, the more destructive pressures involving the acetabulum may force the replacement of the total joint. Radiation therapy has not been used in the treatment of this condition at the University of Miami. PMID- 6289472 TI - Anorexia nervosa: review of current treatment practices. AB - The etiology of anorexia nervosa has defied elucidation for over 100 years and no systematic treatment has yet been developed for the illness. We have reviewed some current findings that relate to both neuroendocrinologic and psychologic defects. Review of current treatment practices reveals that a variety of approaches are used, alone or in combination. Special attention is directed to the rationale for the use of certain neuropharmacologic agents that influence eating behavior. We advocate a flexible, multimodal approach to treatment of this illness, the understanding of which is still in its early stage. PMID- 6289473 TI - Liver tumors and oral contraceptives. PMID- 6289474 TI - Nonbacterial thrombotic endocarditis as a cause of stroke in mitral valve prolapse. PMID- 6289475 TI - [Clinical picture and etiology of serous meningitis in children]. PMID- 6289476 TI - [Possibilities of radionuclide diagnosis of acute myocardial infarct]. PMID- 6289477 TI - BK virus excretion in Fanconi's anaemia. PMID- 6289478 TI - The effects of lithium on the functions of human neutrophils and lymphocytes in vitro and in vivo. AB - The effects of lithium sulphate (LiSO4) at concentrations ranging from 10(-7)M to 10(-2)M on human polymorphonuclear leucocyte (PMNL) and lymphocyte functions in vitro were investigated. The leucocyte function assessed were PMNL motility, post phagocytic hexose-monophosphate shunt activity, myeloperoxidase-mediated iodination of Candida albicans and lymphocyte transformation to mitogens. These same functions as well as the results of serological studies were assessed in normal volunteers prior to ingestion of lithium carbonate (LiCO3), 2 hours and 24 hours after the ingestion of a single oral dose of 480 mg LiCO3, and on the 4th day of ingestion of 2 X 480 mg LiCO3 tablets daily. Incubation of PMNL with LiSO4 at concentrations up to 10(-3)M had no detectable effects on motility or post phagocytic metabolic activity. Higher concentrations (10(-3)M) inhibited these functions. Likewise, at concentrations up to 1 X 10(-4)M LiSO4 had no effects on mitogen-induced transformation of lymphocytes, although higher concentrations did inhibit this activity. These same leucocyte functions were unaffected by ingestion of LiCO3. Levels of serum immunoglobulins and complement components, total haemolytic complement activity and salivary IgA values also remained unaltered. In vitro investigations showed that at a concentration of 10(-3)M LiSO4 had no inhibitory effects on the stimulation of PMNL motility mediated by ascorbate, levamisole and thiamine. PMID- 6289479 TI - [Metastatic calcifications as demonstrated by technetium-99m-pyrophosphate bone scanning. Case reports]. AB - Two cases of chronic renal failure showing very interesting technetium-99m pyrophosphate bone scans are presented. In both cases striking uptake of activity was shown in the left ventricle of the heart, the mucosa of the stomach and in both lungs. This picture was attributed to metastatic calcification in these organs. Cases of metastatic calcification demonstrated with bone-seeking agents have been presented previously, but have mainly shown intense uptake of activity in the lungs and in a few cases, and to a lesser degree, in the stomach. However, we believe that our cases are unique in showing especially the left ventricle of the heart as well as the mucosa of th stomach with exceptional clarity. PMID- 6289480 TI - [Antibodies against cytomegalovirus in blood donors ]. PMID- 6289481 TI - [Pharmacologic and dietary management of prostaglandin synthesis in the prevention of thrombosis]. PMID- 6289482 TI - Computed tomography in the management of soft tissue tumors. AB - Forty-six patients with primary soft tissue sarcomas underwent computed tomographic evaluation. Among 21 patients with retroperitoneal sarcomas, computed tomography was misleading in three patients and accurate and helpful in 18 patients. It was also useful in detecting recurrence by follow-up scanning and determining response to chemotherapy. Among 16 patients with tumors of the extremity, computed tomography demonstrated clinically helpful findings in only two patients and was misleading in two others. Computed tomographic scans were helpful and accurate in evaluating six sarcomas involving the trunk and abdominal wall. Computed tomographic scans were more accurate than ultrasonography or xeroradiography when patients had both imaging modalities. Computed tomographic scans should not supplant clinical examination for tumors of the extremity but are valuable in the preoperative evaluation of sarcomas arising at other sites. They are also valuable in the postoperative follow-up study of patients and may detect a recurrence prior to clinical examination. PMID- 6289483 TI - The Brohee lecture 1982. Gastrointestinal hormones: a field of increasing complexity. PMID- 6289484 TI - Progress in amebiasis. PMID- 6289485 TI - Pathogenic mechanisms and new perspectives in the treatment and prevention of enteric infections. AB - Enteric infections cause more than a billion episodes of diarrhoeal disease in humans each year killing many millions of people, especially young children, in developing countries. Recent progress, reviewed in this article, has enabled that a specific pathogen now can be isolated in the majority of patients with acute diarrhoea, and has also elucidated fundamental pathogenic mechanisms and their pathophysiological effects for several of these agents. Based on this understanding it now seems possible to devise new techniques for the treatment and prevention of diarrhoeal disease to complement those based on fluid replacement therapy and sanitation; prospects for the development of new or improved vaccines, receptor-prophylactic binding agents, and antisecretory drugs are discussed. PMID- 6289486 TI - Hepatic resection for secondary neoplasms. AB - Despite recent advances in chemotherapy, patients with hepatic metastases treated nonsurgically continue to have a dismal survival rate. Using our experience with surgical resection of pulmonary metastases as a model, we have adopted an aggressive surgical approach to secondary hepatic neoplasms. Hepatic resection for metastatic disease in 64 patients undergoing 75 celiotomies resulted in a cumulative survival rate of 45% at 2 years and 34% at 5 years. Operative procedures included 37 wedge resections, 20 segmentectomies, 16 lobectomies, and 2 total hepatectomies in preparation for liver allograft transplantation. The operative mortality rate (less than 30 days) was significant (20%). The most favorable prognosis was associated with Wilms tumor (4 patients) and colorectal carcinomas (29), the 2-year survivals being 80% and 67%, respectively. Long-term survivors include patients with Wilms (17 and 14 years), leiomyosarcoma (12 years), and colorectal (11 and 10 years) primary tumors. Thirty patients with synchronous resections of the primary tumor and the hepatic resections had 2- and 5-year survival rates of 29% and 26%, whereas 34 patients with metachronous resections had survival rates of 64% and 30%. Eighteen patients underwent resections of multiple hepatic metastases with a survival rate essentially no worse than that of patients with isolated metastases, but careful case selection may have played an important role in the outcome. The extent of hepatic resection was primarily dictated by location and number of metastases. More extensive resections were not associated with a higher long-term survival rate but did have a higher operative mortality. We conclude that an aggressive surgical approach in the treatment of metastatic disease confined to the liver offers a chance for long-term survival and significant palliation in selected patients whose primary tumor is controlled. PMID- 6289489 TI - [Endocrine pancreas tumors]. PMID- 6289488 TI - Perchloric acid treatment of human blood for quantitative endotoxin assay using synthetic chromogenic substrate for horseshoe crab clotting enzyme. AB - A new reliable human blood treatment was established for quantitative endotoxin assay using synthetic chromogenic substrate [Boc-Leu-Gly-Arg-rho-nitroanilide]. Addition of perchloric acid in a final concentration of 1.25% to platelet-rich plasma or serum in a 2:1 volume ratio completely eliminated nonspecific amidase activities as well as inhibitors. By this method, the recovery of added endotoxin was nearly 100%, and was almost independent of sample dilutions and anticoagulants. PMID- 6289487 TI - [Indications for immunosuppressive treatments and for plasma exchange in inflammatory neuropathies]. PMID- 6289490 TI - A new Epstein-Barr virus negative Burkitt's lymphoma derived cell-line. I. Analysis of cell surface markers and abnormal expression of HLA antigens. AB - CHEV, a new Epstein-Barr virus negative Burkitt's lymphoma derived cell line has been studied. Karyotype analyses demonstrated the t (8; 14) characteristic translocation. Cell surface characterisation of this line showed the presence of mu and chi immunoglobulin chains and beta 2-microglobulin and the absence of the complement receptor. We are unable to detect HLA--A, B alloantigens on the cell surface which was in contrast with an apparently normal expression of these antigens in the cytoplasm. PMID- 6289491 TI - Liver microsomal enzyme induction and toxicity studies with 2,4,5,3',4' pentabromobiphenyl. PMID- 6289493 TI - Influence of age and nephrectomy on metastatic patterns in Wilms' tumors. AB - Data from 194 autopsy cases of Wilms' tumor were analyzed statistically to investigate the mode of dissemination by classifying them according to the number of organs involved in metastasis, age and history of nephrectomy. Metastasis-free cases were more frequent in cases under two years of age, in bilateral cases, and in non-nephrectomized cases. The frequencies of metastasis to various organs were essentially similar among the various age groups, except those to the contralateral kidney and intestines in cases under two years of age, which were significantly more frequent. Nephrectomized cases showed a significantly higher incidence of metastasis to the peritoneum than non-nephrectomized cases. In cases with metastasis to one organ, metastasis to the lungs, lymph nodes and liver showed low frequencies (17%, 13% and 17%, respectively), but that to the peritoneum showed a rather high frequency (21%). Radiation therapy for the peritoneum after nephrectomy is recommended. PMID- 6289492 TI - Synergistic toxicity of carbon tetrachloride and several aromatic organohalide compounds. AB - Subacute (20 days) oral administration of hexachlorobenzene (HCB) or the organohalide mixtures polybrominated biphenyls (PBB) or polychlorinated biphenyls (PCB) greatly increased th susceptibility of male rats to the toxic effects of carbon tetrachloride (CCl4). CCl4-induced acute growth retardation, renal tubular functional impairment and hepatocellular necrosis were quantitatively greater in rats pretreated with the aromatic organohalides than in naive rats. Pretreatment with HCB, PBB or PCB also reduced survival after i.p. administration of CCl4 and increased the severity of morphological liver injury. Despite functional impairment, only minor histological alterations attributable to CCl4 were detected in the kidney. CCl4 as well as HCB, PBB or PCB increased the lipid content of the liver, but not the kidney. CCl4 administration depressed energy dependent accumulation of organic ions by renal cortical slices in vitro in a dose-dependent manner and increased the basal rate of respiration of renal cortical tissue in vitro. It is concluded from these results that exposure to certain commercial aromatic organohalides can greatly alter organ response to toxicants such as CCl4. PMID- 6289494 TI - Differences of microviscosity and lipid composition in normal human T and B lymphocytes. AB - Human T and B lymphocytes differ morphologically and immunologically although little is known concerning their physiological parameters and biochemical constituents. In this study, microviscosity and lipid composition in normal human B and T lymphocytes were investigated. Microviscosity was examined using 1,6 diphenyl-1,3,5-hexatriene as a probe and lipid composition was analyzed by thin layer chromatography with flame ionization detection. Microviscosity of T lymphocytes was higher than that of B lymphocytes throughout the temperature ranges tested, and microviscosity of whole lymphocytes was similar to that of B lymphocytes rather than that of T lymphocytes. There was no significant difference in lipid composition between B and T lymphocytes. However, it was postulated that the concentration of lipid per area of plasma membrane in B lymphocytes might be less than that in T lymphocytes. It was concluded that B lymphocytes show a less rigid membrane fluidity than T lymphocytes because of the lower concentration of lipid per area of plasma membrane. PMID- 6289495 TI - Surgical approaches to orbital lesions. PMID- 6289496 TI - Tumors of the lacrimal gland and lacrimal sac. PMID- 6289497 TI - HLA-DR typing of fetal human spleen and liver lymphoblastoid cells transformed by Epstein-Barr virus. AB - Because few HLA-DR-positive cells are present in the fetal spleen and liver, full HLA typing cannot be performed. However, B lymphocyte precursors can be transformed with Epstein-Barr virus to produce lymphoblastoid cells which express HLA-A, B, and DR antigens. Successful transformation was achieved, usually with spleen and liver, in nine fetuses aged from 15 to 18 weeks, mostly within 7 to 14 days of initiation of the cultures. Spleen-derived lymphoblasts were more suitable for typing because of their greater homogeneity and higher viability. Tissues from two 13-week fetuses from prostaglandin-induced abortions and from a spontaneously aborted 22-week fetus could not be transformed. This is probably attributable to prolonged ischemia before the tissues were obtained but, in the 13-week fetuses, absence of B lymphocyte precursors was not excluded. HLA-DR typing may be useful in obtaining well matched donor-recipient pairs in fetal pancreatic islet transplantation. PMID- 6289498 TI - [Aconitine modification of cell membrane sodium channels of a neuroblastoma]. AB - Currents through normal and aconitine-modified sodium channels in the perfused neuroblastoma cell are measured under voltage clamp conditions. Aconitine shifts the voltage range of activation of the sodium channels towards more negative potentials by about 20 mV, and changes the selectivity, so that channels become more permeable to NH4+ than to Na+ ions. The currents through aconitine--modified channels are inactivated almost completely like those through the normal ones. Aconitine is effective when applied to both sides of the cell membrane. Steady state characteristics of gating are discussed in terms of the model assuming three main states of the gate machinery: closed, open and inactivated. PMID- 6289499 TI - [Characteristics of "photo depletion" of the green fluorescence of tumor cells]. AB - The green (oxidized flavoproteins) fluorescence intensity was found to increase during investigation of NADH and oxidized flavoproteins fluorescence with the use of optimal excitation of different fluorescence bands. This effect was observed under excitation with blue light (436 nm). It is suggested that in some malignant cells, the structure of flavoproteins (probably of mitochondrial ones) may be altered in the way of increasing the quantum yield under the action of light irradiation. PMID- 6289500 TI - Recognition of pseudocowpox in Nigeria. PMID- 6289501 TI - Degree of malignancy of maxillary sinus cylindromas in relation to histologic characteristics. PMID- 6289503 TI - Adenoid cystic carcinoma of female periurethral area. Light and electron microscopic study. PMID- 6289502 TI - The outcome of Wilms' tumor in infants. Italy 1970-79. AB - Thirty-four infants under 1 year of age with Wilms' tumor were diagnosed and treated in 14 Italian pediatric oncology units during 1970-79. The 3-year survival rates decreased with higher group unilateral tumors: 95% in group I Wilms' tumor, 75% in group II and 20% in group III. The survival rates for children with group I and II Wilms' tumor were similar for those who were treated with surgery and chemotherapy and those who also received postoperative radiotherapy. During 1975-79 fewer patients with group I Wilms' tumor received radiotherapy (1 of 11) than during 1970-74 (4 of 6, p less than 0.05). All these children are alive at this writing. PMID- 6289504 TI - [Acute abdomen syndrome in rupture of a kidney tumor in children]. PMID- 6289505 TI - [Effect of the morphologic structure and degree of tumor differentiation on the survival of lung cancer patients]. PMID- 6289506 TI - Experimental louping-ill virus infection in two species of British deer. PMID- 6289507 TI - Infectious bronchitis haemagglutination inhibition test. PMID- 6289508 TI - Safety and efficacy of live and inactivated infectious bovine rhinotracheitis vaccines. AB - Two commercial live virus infectious bovine rhinotracheitis (IBR) vaccines for intranasal administration and an inactivated polyvalent calf pneumonia vaccine were compared for safety and efficacy in calves against experimental IBR infections. All three products were clinically safe for use in young calves; a mild, transient, febrile response was induced by one of the live vaccines. Vaccinal virus was recovered for up to 16 days after vaccination from nasal secretions of all calves given live vaccine. Both live vaccines stimulated a serum neutralising antibody response, but the inactivated vaccine failed to elicit any serological response. Following intranasal challenge four months after the first dose of vaccine, all live virus vaccinates remained systemically healthy. A slight nasal discharge and a few rapidly healing ulcers of the nasal mucosa were the only abnormalities observed. Both the group given the inactivated vaccine and the unvaccinated controls developed clinical IBR with pyrexia, ocular and nasal discharges, severe ulceration of the nasal mucosa and tracheitis and tachypnoea to varying degrees of severity. Parenteral administration of dexamethasone six months after challenge induced reactivation of virus shedding followed by a rise in humoral antibody titre irrespective of the original vaccination history. PMID- 6289509 TI - Transfer of embryos from bovine leukaemia virus-infected cattle to uninfected recipients: preliminary results. AB - One hundred and fifty-one, day 6 or 7, embryos collected from cattle infected with bovine leukaemia virus (BLV) were transferred to uninfected recipients. Thirty-two pregnancies resulted. Two animals aborted at seven months. Three sets of twins and one single calf were still-born. The remaining 26 pregnancies produced 27 live calves which were raised to six months of age. All of the recipients, pregnant and non-pregnant, and all of the calves remained serologically negative for antibodies to BLV-glycoprotein antigen. PMID- 6289510 TI - Bluetongue in western Turkey. AB - In October 1977 clinical bluetongue broke out in Aydin province, western Turkey and spread to adjacent provinces in the autumn months of 1978 and 1979. The outbreak was caused by a virus of serotype 4 and appeared to occur in a totally susceptible population. It was eventually controlled by widespread use of attenuated type-specific vaccine. Bluetongue virus was isolated from sheep on several occasions and also from a calf with congenital arthrogryposis and hydranencephaly. This latter finding is discussed in relation to Akabane virus, a recognised arbovirus teratogen thought to be present in the same area at the same time. PMID- 6289511 TI - Incidence and histological structure of the storiform pattern in benign and malignant fibrous histiocytomas. AB - A starlike arrangement of cells and fibers, the "storiform pattern", was found to be a typical, but not obligatory, histological feature of benign and malignant fibrous histiocytomas. In 155 benign fibrous histiocytomas storiform structures were missing in 29 cases, chiefly of the fibroblastic type comparable with classical "dermatofibroma". 12 of 70 malignant fibrous histiocytomas did not reveal storiform structures, especially the cellular pleomorphic variant, i.e. the classical pleomorphic sarcoma. Storiform structures were either small and highly cellular with few fibers (collagen type III), or larger, less cellular, but with abundant fibers (collagen type I). There was no sharp demarcation between these two extremes, but many transitional structures or patterns were seen. The histiocytic nature of the cells was demonstrated in both variants of storiform structures by immunhistochemical methods on paraffin embedded material. Alpha 1-antichymotrypsin was especially valuable in this respect. PMID- 6289512 TI - Cellular haemangioma. Light and electron microscopic studies of two cases. AB - Light and electron microscopic studies were conducted on the immature vascular tumors of two infants, containing various stages of differentiation of the blood vessels and both benign haemangioendotheliomas and haemangiopericytomas. We were able to confirm the existence of two kinds of hyperplastic, immature cells i.e. endothelial cells and pericytes in the same tumor. Presence of crystalloid inclusions in the endothelial cells and absence of the Weibel-Palade bodies, as well as a deficiency in factor VIII-related antigen and no tissue fibrinolytic activity, suggested that the endothelial cells in these lesions were immature. Electron microscopic studies appear more decisive in the diagnosis of heterogenous cellular vascular tumors than light microscopy and if available should be used to aid in the final diagnosis. The authors propose that the term cellular haemangioma would be more appropriate in describing this vascular entity. PMID- 6289513 TI - Primary malignant mixed mesodermal tumor of the gallbladder. Report of a case and critical review of diagnostic criteria. AB - A rare case of primary malignant mixed mesodermal tumor of the gallbladder arising in a 75 year old woman is reported. The previously published cases of similar tumors were reviewed in order to outline the histological features and the histogenesis. In diagnosing a malignant mixed mesodermal tumor of the gallbladder it was imperative that we excluded malignant neoplasms with multiple histological patterns. The diffuse and close intermingling of the epithelial and mesenchymal component ruled out a collision tumor. The high mitotic rate, the typical reticulin pattern and the obviously malignant osteoblasts excluded a spindle cell carcinoma with osseous metaplasia. The authors conclude that this is the first case of malignant mixed mesodermal tumor with evident osteosarcomatous areas, described in the gallbladder. PMID- 6289514 TI - Neurohormonal peptides and argyrophil cells in an androblastoma of the ovary. AB - An ovarian epithelial tumour is described with a compound histological structure containing features of an androblastoma with well differentiated Sertoli-Leydig cell structures and features resembling a carcinoid tumour. Scattered argyrophil cells were demonstrated in both components of the tumour. Neurohormonal peptides were found immunocytochemically in a minor fraction of the argyrophil cells. These included pancreatic polypeptide (PP), glucagon/enteroglucagon and enkephalin. These findings indicate a high degree of multipotentiality of the progenitor cells of the neoplasm. PMID- 6289515 TI - Structural characterization of the adenovirus 18 inverted terminal repetition. PMID- 6289516 TI - Nearest neighbor relationships of major structural proteins within bovine leukemia virus particles. PMID- 6289517 TI - Structural similarities of proteins encoded by three classes of avian sarcoma viruses. PMID- 6289518 TI - Use of transcription probes for genotyping rotavirus reassortants. PMID- 6289520 TI - Kirsten murine sarcoma virus-coded p21ras may act on multiple targets to effect pleiotropic changes in transformed cells. PMID- 6289519 TI - The nuclear matrix is involved in herpes simplex virogenesis. PMID- 6289521 TI - Regulation of polyoma virus early transcription in transformed cells by large T antigen. PMID- 6289522 TI - Characterization of cytomegalovirus immediate-early genes. I. Nonpermissive rodent cells overproduce the IE94K protein form CMV (Colburn). PMID- 6289523 TI - The effect of the vesicular stomatitis virus-associated protein kinase on viral mRNA transcription in vitro. PMID- 6289524 TI - Varied responses of human B-lymphoblastoid cell lines to infection with vesicular stomatitis virus. PMID- 6289525 TI - Restriction endonuclease analysis of variants of phage T1: correlation of the physical and genetic maps. PMID- 6289526 TI - [Role of food fiber in nutrition]. AB - Numerous experiments indicate that in the pathogenesis of so-called civilization diseases, low consumption of dietary fibers seems to be one of etiological factors. Therefore, it is not surprising that dietary fibers play an ever increasing role both in prevention and dietetic management of the diseases. Among these fibers wheat bran containing about 50% of dietary fibers has the most beneficial physiological effect. Besides, wheat bran is fit very well for production of different foodstuffs. Data are reported on the content of crude and dietary fibers in cabbage, radish, carrot, wheat bran and brown bread. PMID- 6289527 TI - [Specific class M immunoglobulins in viral hepatitis A patients (their diagnostic and prognostic significance)]. AB - The study included 119 children and 138 adults suffering from viral hepatitis A. The controls consisted of 114 adult patients with hepatitis B. Examinations of the total antibody to hepatitis. A virus revealed a rise in titres from 1 : 450 in the first days of the disease to 1 : 4 400 in the convalescent period. In the acute period the antibodies were represented by immunoglobulins M which circulate in the blood up to 4-5 months of convalescence. At 3-4 weeks after the onset of the disease, synthesis of immunoglobulins G begins which are anamnestic antibodies. The importance of specific immunoglobulins M in virus elimination from the host and their role in the early diagnosis of the disease have been demonstrated. PMID- 6289528 TI - [Determination of hepatitis A virus in the feces of patients in the dynamics of the infectious process]. AB - Excretion of hepatitis A virus (HAV) in the time course of the infectious process is described. Among 197 fecal specimens collected from 45 patients HAV was found in 40 (20.5%); in 51% of the patients the virus was isolated in the 1st week of the icteric period, in 25% in the 2nd week and in 4% in the 3rd week. The buoyant density in cesium chloride gradient of virus specimens obtained on the 1st-2nd day of jaundice was represented by one peak of 1.30-1.35 g/cm3, whereas the material collected at late periods of the disease had heterogeneous buoyant density in zones from 1.24 to 1.43 g/cm3. The immune electron microscopic studies showed the fecal specimens collected early in the disease to be richer in virus than those collected at later intervals. PMID- 6289529 TI - [Heterogeneity of virus-specific flavivirus proteins]. AB - The polyacrylamide gel analysis of large intracellular virus-specific proteins NV5, NV4, and the intracellular form of structural protein V3 established differences in the electrophoretic mobility of each of these proteins formed in cells infected with tick-borne encephalitis, Powassan, Langat, and West Nile viruses. It is assumed that these differences in the electrophoretic mobility of NV5, NV4 proteins, and the intracellular form of V3 protein reflect the differences in the primary structure of each of these proteins in the viruses examined. PMID- 6289531 TI - [Typing of herpes simplex virus strains by analyzing the viral DNA using restriction endonucleases]. AB - Analysis of the products of viral DNAs cleavage by restrictive endonucleases showed the strains of herpes simplex virus Us and L2 used for herpes vaccine manufacture to belong to type 1 and the VN strain to type 2 of herpes simplex virus. This method is found to be optimal for typing of herpes simplex virus strains. PMID- 6289530 TI - [Humoral immunity to herpes simplex virus type 2 in cervical cancer]. AB - The analysis of the results of serological studies revealed differences between the number of positively reacting sera from patients with cervical carcinoma and from normal women. The activity of complement-fixing and virus-neutralizing antibodies in the patients' sera and the frequency of their detection were higher than in the sera from normal subjects. These results in combination with other methods of laboratory examinations may be used for early detection of cervical carcinoma as well as for determination of high risk groups with regard to oncological diseases. PMID- 6289532 TI - [Circular dichroism study of the ribonucleoprotein structure of Sendai virus isolated from infected chick embryo cells]. AB - Thermostability and secondary structure of RNA, a component of ribonucleoprotein (RNP) of Sendai virus isolated from infected chick embryo cells (cRNP) and from virions (vRNP) were studied by the method of circular dichroism. The secondary structure of RNA in cRNP was shown to be practically no different from that of free RNA whereas incorporation of RNA into vRNP was accompanied by a significant decrease in the number of paired bases in it. Comparison of thermodenaturation parameters of cRNP and vRNP also revealed significant differences in their structural organization. Thus, melting of cRNP as well as of free RNA is of markedly non-cooperative nature indicating poor RNA-protein interactions in the complex. In contrast, the process of vRNP thermodenaturation occurs in a step wise manner in a narrow temperature range indicating a significant role in the maintenance of this RNP structure of both RNA-protein and, apparently, protein protein interactions. PMID- 6289533 TI - [Solubilization of myxovirus surface glycoproteins by using a nonionic detergent octylglucoside]. AB - Using alpha- and beta-forms of the non-ionic detergent octyl glucoside the conditions have been developed for treatment of influenza virus and Sendai virus suspensions with the purpose of solubilization from virions of exclusively surface glycoproteins. Both alpha- and beta-forms of the detergent have been shown to have similar effectiveness for the recovery of chromatographically pure preparations of surface glycoproteins. PMID- 6289534 TI - [Isolation of the Tribec virus in Byelorussia]. PMID- 6289535 TI - [Polyradiculoneuritis with concomitant arrhythmia caused by Coxsackie B virus]. PMID- 6289536 TI - [Small intestine cancer as a cause of acute intestinal occlusion]. PMID- 6289537 TI - [Complete triploidy (69,XXX) surviving until the age of 7 months]. AB - A patient with complete triploidy surviving until the age of 7 months is described. The diagnosis was established by the observation of 69,XXX chromosomes in lymphocyte and fibroblast cultures. Our findings suggest two active X chromosomes, since only one Barr body was found in buccal smear and drumsticks were within the normal range for females. Clinical findings disclosed a left sided hemihypertrophy. There were several degenerative stigmata such as craniofacial dysmorphism, hypertelorism, low-set malformed ears and a high-arched palate. The CNS malformation consisted of microcephaly, diffuse cortical atrophy and a cavum septi pellucidi. There was soft tissue syndactily of all extremities and polydactily of the right lower extremity. Psychomotor development was severely retarded. Our patient presented with severe convulsions, retardation and recurrent bacterial infections, particularly of the respiratory tract. The girl died suddenly at the age of 7 months fulminating bilateral pneumonia. Permission for autopsy could not be obtained. PMID- 6289539 TI - [Chronic constipation and the abuse of laxatives]. PMID- 6289538 TI - Bronchoplasty for bronchogenic carcinoma. PMID- 6289540 TI - [Development and use of radio- and enzyme immunoassays in microbiological and immunological diagnosis. III. Comparative studies on viral antibody determination with hemagglutination, radio- and enzyme immunoassays]. PMID- 6289541 TI - [Experimental studies of the circulation of enteropathogenic viruses]. PMID- 6289542 TI - [Use of passive hemagglutination in the diagnosis of mumps virus infections]. PMID- 6289544 TI - [Virus-induced changes in heart and skeletal muscle]. PMID- 6289543 TI - [Current problems of visual-vestibular interaction]. PMID- 6289545 TI - [Viral infections of the hematopoietic system: Epstein-Barr-virus and other less characterized viral infections]. PMID- 6289546 TI - [Lymph node lesions caused by virus infections]. PMID- 6289548 TI - [Rapid diagnosis of herpes simplex encephalitis (HSE) using the immunoperoxidase method]. PMID- 6289547 TI - [Lymphoepithelial carcinoma of the nasopharynx: a special tumor?]. PMID- 6289549 TI - [Investigations to demonstrate latent viral infection of varicella-Zoster-virus in human spinal ganglia]. PMID- 6289550 TI - [Discussion of possible pathogenetic relations between virus infection, immune complex vasculitis, and chronic graft versus host reaction in bone marrow transplant recipients]. PMID- 6289551 TI - [Proliferation kinetics of a virus-induced sarcoma in rats]. PMID- 6289552 TI - [Cross-reaction of human breast cancers with the envelope glycoprotein gp 52 of the mouse mamma tumorvirus (MMTV)]. PMID- 6289553 TI - [Electron microscopical observations of the junctional elements in virus producing murine mammary tumors]. PMID- 6289554 TI - [Virus infections and their effect on development in growth]. PMID- 6289556 TI - [Intrauterine coxsackie B infection in arthrogryposis multiplex congenita syndrome]. PMID- 6289555 TI - [Possibilities and limits in the detection of a viral pathogenesis in diagnosis and research. Electron microscopy]. PMID- 6289557 TI - [Observations in transplacental infection with H-SE polyoma virus in inbred Lewis rats]. PMID- 6289558 TI - [Differences in morphology of polyoma H-SE induced tumors in fetal and newborn rats]. PMID- 6289559 TI - [Virus infections of the skin]. PMID- 6289561 TI - [The use of nucleic acid hybridization exemplified by Epstein-Barr virus associated diseases]. PMID- 6289560 TI - [Virus and skin. Dermatological and pathomorphological aspects]. PMID- 6289563 TI - Survival after complete excision of renal vein of a solitary kidney. PMID- 6289562 TI - [Molecular pathogenetic mechanisms and immunity in candidiasis]. PMID- 6289564 TI - [Comparison of antimicrobial activity and stability to beta-lactamases of cefoperazone, cefotaxime, lamoxactam and ceftriaxon]. AB - Ceftriaxon (Ro 13-9904) is a new cephalosporin, whose structure resembles cefotaxime. This study compares the minimal inhibitory concentrations (MIC-s) of Ceftriaxon, cefoperazone, cefotaxime, lamoxactam and gentamicin against 622 isolates of various Enterobacteriaceae, non-fermenters and Aeromonas hydrophila. Furthermore the minimal bactericidal concentrations (MBC-s) were determined for some organism; additionally destruction of beta-lactamantibiotics in the presence of beta-lactamases was studied by a bioassay technique. The in vitro activity of Ceftriaxon against gramnegative bacteria was found very similar to that of cefotaxime. The susceptibility of Enterobacter sp. to the three cephalosporines was exceeded by lamoxactam while cefoperazone was the most active beta-lactam antibiotic against Pseudomonas. Lactamases, which inactivated cefotaxime also destroyed Ceftriaxon in a greater extend. No hydrolysis of lamoxactam and cefoperazone occurred by nearly all bacterial extracts tested. PMID- 6289565 TI - Bacteriological investigations with beta-lactamase inhibitors. AB - MIC's of beta-lactamase producing strains of P. aeruginosa, E. coli, P. vulgaris, P. rettgeri, E. cloacae, K. pneumoniae, S. marcescens are determined against piperacillin and in the case of S. aureus against amoxycillin and penicillin G. All the strains studied were resistant to these antibiotics. Their MIC's were determined in the combination with different concentrations (2, 5, 10 mg/l) of clavulanic acid and penicillanic acid sulfone respectively. The combination of piperacillin and clavulanic acid acts synergisticly against Klebsiella sp., indolpositive Proteus sp., and E. coli. Penicillanic acid sulfone was less active than clavulanic acid apart from indolpositive Proteus sp. Against S. aureus however both inhibitors showed the same synergistic efficacy. In detail both inhibitors are very active against constitutive penicillinase, they also exhibited a good activity against Type OXA-2, OXA-3 and SHV-1 beta-lactamase; they were less active to Type TEM-1 and OXA-1 enzyme and did not work against Type Ib, TEM-2 and Type IV beta-lactamase. In investigations using the biophotometer it could be demonstrated that the beta-lactamase inhibitor and the penicillin should be given simultaneously in order to obtain an optimal synergistic effect. PMID- 6289566 TI - Scanning electron microscopical investigations on the respiratory epithelium of the Syrian golden hamster. IV. In vitro effects of dimethylsulphoxide and benzo(a)pyrene. AB - The biological effects of in vitro exposure of cultured fetal hamster tracheal epithelium to benzo(a)pyrene in 0.5% dimethylsulphoxide (BaP: 5 and 10 micrograms/ml medium) were studied by light and scanning electron microscopy. The solvent, DMSO, alone induced a slight, temporary retardation in development and differentiation of cilia. At day 28 of culture, however, differences in growth and maturity disappeared between DMSO-exposed and untreated explants. No dose related differences found in the surface morphology of alterations were seen in BaP-treated tracheae; the explants revealed metaplastic foci and reduced, incomplete ciliogenesis compared to DMSO- and non-treated controls. Towards the end of the culture period, ciliogenesis tended slowly to replace epithelial alterations. PMID- 6289567 TI - [Neural therapy and accupuncture in gynaecology and obstetrics]. AB - A brief characterisation is made of the working principles underlying neural therapy under local anaesthesia or accupuncture. Common approaches to therapy are offered by disorders of autonomous regulation, including inflammatory processes, and by purely functional disorders.--There are many applications in gynaecology and obstetrics. A brief statistical information on lumbosacral pain is quoted as an example. Optimum performance can be expected from them, when used in combination with proven therapeutic methods. They provide a low-cost approach to reducing both the consumption of antibiotics and other pharmaceuticals as well as time of morbidity. PMID- 6289568 TI - [Gestational luteoma]. PMID- 6289569 TI - Transient viraemia with bovine leukaemia virus in bulls. PMID- 6289570 TI - [Genetics of staphylococcal plasmids]. PMID- 6289572 TI - [Study of the DNA of the causative agent of whooping cough, Bordetella pertussis]. AB - A number of physico-chemical properties of Bordetella pertussis DNA has been studied. The values of its floating density and melting point have been established, which has allowed one to calculate the GC composition of B. pertussis DNA. The average molecular weights of the fragments of B. pertussis DNA, resulting from its hydrolysis with specific endonucleases EcoR, BamH 1, Sal 1, Pst 1, have been determined, and thus the basis has been provided for establishing the number of clones necessary for obtaining the complete lset of B. pertussis genes. PMID- 6289571 TI - [Carbamoyl phosphate biosynthesis in Streptococci]. AB - Nine streptococcal strains belonging to different serological groups (A, B, C, D) were shown to synthesize carbamoyl phosphate from ammonium hydrocarbonate and ATP. The reaction was catalyzed by carbamate kinase (EC 2.7.2.2). The speed of the reaction was evaluated according to the increase of the content of citrullin (the combination of carbamate kinase and ornithine transcarbamoylase). The representatives of different serological groups were found to have quantitative differences in carbamate kinase activity: the highest specific activity (13 nmol of citrullin per minute in 1 mg of dried microbial biomass) was detected in group A streptococci, while group D streptococci showed the lowest specific activity (0.5 nmol). PMID- 6289573 TI - [Analysis of insertions of the determinant of drug resistance to kanamycin and chloramphenicol into bacteriophage lambda att80]. AB - The insertion sites of elements Tn9 and Tn601 which determine chloramphenicol and kanamycin resistance have been detected restriction analysis. The functioning of transposons i.e. their stability or instability, has been found to influence the specificity of their insertions into the genome of lambda att80 bacteriophage. During transposition from stable integration sites both transposons are inserted into the regions of the lambda att80 bacteriophage genome, definite for each transposon. However, during transposition from the site of unstable integration both determinants of drug resistance are inserted into different regions of the phage genome. PMID- 6289574 TI - [Enzymes involved in energy metabolism in the brains of schizophrenic patients (electron-cytochemical study)]. PMID- 6289575 TI - ACTH and brain membrane phosphorylation: a model for modulation by neuropeptides. AB - It has been hypothesized that changes in the phosphorylation of synaptic membrane constituents (proteins and lipids) may affect transmission in certain types of synapses. In this paper some of the recent evidence that neuropeptides like ACTH may bring about their behavioral activity by influencing brain protein and lipid phosphorylation is reviewed. An ACTH-sensitive, cAMP-independent protein kinase was isolated from rat brain synaptosomal plasma membranes. This enzyme was partially characterized and it was observed that its activity greatly depended on the presence of calcium ions. One of its substrate proteins B-50 (MW 48,000; IEP 4.5) may play a key role in the turnover of a special class of membrane phospholipids i.e. the (poly)phosphoinositides. Evidence was obtained to suggest that the degree of phosphorylation of the B-50 protein determines the conversion of diphosphoinositol to triphosphoinositol. A model which links the protein phosphorylation to lipid phosphorylation and which points to a functional role for peptides in the regulation of the permeability of brain membranes for calcium ions will be discussed. As the structure-activity relationship for the peptide effects on grooming behavior closely resembles that on phosphorylation, it is assumed that this neurochemical event may indeed be of relevance to the biological activity of the peptide. As the ion permeability may be altered by the peptide it can be suggested that this may lead to modulation of transynaptic information processing in the brain. PMID- 6289576 TI - Action of a calcium blocking agent (D600) on electrogenesis in the normal heart and the ischaemic heart in the dog. PMID- 6289577 TI - Relationship between cytomorphologic features and prognosis in small-cell carcinoma of the lung. AB - Small-cell bronchial carcinoma cells obtained from 18 patients by fiberbronchoscopic brushing were examined to determine whether the cytomorphologic features could be related to the prognosis. Nuclear features such as small (less than 10 micrometer) or large (greater than 11 micrometer) size, round or aberrant shape and dark or light staining, were evaluated. In small-cell carcinoma cases with a very poor prognosis (death within four months), the incidence of small, round, darkly stained nuclei was much greater and a coarse or pyknotic chromatin pattern was more frequent in comparison with small-cell carcinoma cases with a better prognosis (survival greater than two years). In cases with a good prognosis, the incidence of large, lightly stained nuclei with aberrant shapes was much greater and a fine chromatin pattern was more frequent in comparison with cases with a very poor prognosis. Pertinent cytomorphologic features thus seem to be related to the prognosis. PMID- 6289578 TI - Malignant fibrous histiocytoma. Cytologic, light microscopic and ultrastructural studies. AB - Studies of the cytologic, light-microscopic and electron-microscopic features of three malignant fibrous histiocytomas (MFH) confirmed the presence of histiocytelike and fibroblastlike cells. The ultrastructural features suggest that the neoplasm possibly originates in undifferentiated mesenchymal cells that are capable of differentiating into histiocytic and fibroblastic cells. Cytologic features seen in smears obtained by thin needle aspiration included loose aggregates of markedly pleomorphic, elongated cells, histiocytelike cells and isolated bizarre giant cells. Phosphotungstic acid hematoxylin (PTAH) and oil red O staining of air-dried smears were helpful in differentiating MFH from pleomorphic variants of liposarcoma and rhabdomyosarcoma. PMID- 6289580 TI - Cytopathologic aspects of a metastatic malignant mixed Mullerian tumor of the uterus. Report of a case with transabdominal fine needle aspiration biopsy. AB - The cytologic details of a fine needle aspiration biopsy of an intraabdominal metastatic malignant mixed Mullerian tumor of the uterine corpus are presented. Cytologically, the malignant stromal and epithelial elements resembled those in previous histologic materials. Given the aggressive nature of this lesion, the patient may initially present with an inoperable tumor; cytologic examination of an aspiration biopsy may be diagnostic without the need for an exploratory laparotomy. PMID- 6289579 TI - Cytology of synovial sarcoma metastases in pleural fluid. AB - Report is made of the cytology of pleural fluid arising from extensive pleuropulmonary metastases of a biphasic synovial sarcoma of the leg of a 35-year old male. Both fibrous, spindle-shaped and epithelioid types of tumor cells were present. The epithelioid cells were seen in groups or arranged in glandlike structures. Transformation of the fibrous, spindle-shaped tumor cells into the epithelioid ones was observed, suggesting that these two types of tumor cells are cytogenetically related. PMID- 6289581 TI - Endometrial cytologic sampling: utility of the Mi-Mark Endometrial Sampler. PMID- 6289582 TI - Aspiration cytology of oat-cell carcinoma metastatic to the parotid gland. PMID- 6289584 TI - Effects of diets rich and poor in fibres on the development of hereditary diabetes in mice. AB - Young mice with different genetic dispositions to obesity and hyperglycaemia were fed isocaloric diets rich (B-diet) or poor (O-diet) in fibre from rye. Normal C57BL/KsJ=+/+-mice developed equally well on the two diets with no differences in serum glucose or insulin. In non-inbred ob/ob-mice both diets were as effective as standard laboratory diet in supporting the fat accumulation and weight gain typical of these animals. No effect on serum insulin was observed, but at the age of 10 and 13 weeks the ob/ob-mice were more hyperglycaemic on the O-diet than on the B-diet. C57BL/KsJ-db/db-mice on the B-diet developed and maintained obesity until 20-25 weeks of age; they then began to lose weight and died at a median age of 36 weeks. KsJ-db/db-mice on the O-diet soon began to mobilize their fat reserves and died at a median age of 28 weeks; they also exhibited a more severe hyperglycaemia and less pronounced hyperinsulinaemia than KsJ-db/db-mice fed the B-diet. It is concluded that, at least in mice, the manifestations of a defined genetic disposition to severe diabetes mellitus can be modified by such simple an environmental factor as rye bran. PMID- 6289583 TI - Restoration of oestrogen-positive feedback effect on LH release in women with prolactinoma by transsphenoidal surgery. AB - The effects of hyperprolactinaemia on the hypothalamo-pituitary axis by iv injection of 100 micrograms luteinizing hormone releasing hormone (LRH) in 7 women with prolactinoma before and 3 months after normalization of the Prl level by transsphenoidal surgery. A dose of 20 mg of conjugated oestrogen (Premarin) was also infected iv into patients with prolactinoma before and 4 months after surgery, and the serum LH levels were determined serially for 120 h after the injection. Surgical treatment caused significant reduction of the mean (+/- SE) serum prolactin (Prl) level from 123.3 +/- 7.8 to 19.4 +/- 5.6 ng/ml. But the differences in the basal levels of HL (11.3 +/- 2.2 to 8.6 +/- 1.5 mIU/ml), FSH (8.3 +/- 2.4 to 10.6 +/- 3.7 mIU/ml) and oestradiol (26.6 +/- 8.6 to 37.5 +/- 5.5 pg/ml) before and 4 months after surgery were not significant. An exaggerated LH response to LRH in untreated prolactinoma patients was also observed after surgical treatment. After surgical treatment, patients showed LH release with a peak between 48 and 72 h after the injection of Premarin, whereas before treatment they did not show any LH discharge. The mean percent increase in LH between 48 and 72 h was also significantly higher after operation than before operation. These results suggest that the hyperprolactinaemia in prolactinoma patients may cause an impaired positive feedback effect of oestrogen on LH release and that this derangement can be reversed by reduction of the Prl level by adenomectomy. PMID- 6289585 TI - Studies on the mechanism by which ACTH stimulates renin activity and angiotensin II formation in man. AB - Ten IU of ACTH (1-24) per day was infused for 34 h (starting at 7 a.m.) into 8 normal men on a constant diet containing 135 mM Na+ per day. All subjects retained between 152 and 181 mM of sodium. Potassium balance was negative. Plasma renin activity (PRA) and plasmaangiotensin II (P-A-II) started to rise in most subjects after 6 to 8 h of infusion, reached a maximum after 24 h and then tended to decline. As shown previously, the rise in PRA is not due to a rise in plasma renin substrate concentration. Systolic, but not diastolic blood pressure increased significantly on the second day of ACTH-infusion. Plasma cortisol (P-F) was continuously stimulated by ACTH. Plasma aldosterone (P-aldo) increased rapidly 1 h after ACTH administration, then tended to fall, and increased again in most subjects, roughly in parallel with PRA. No significant changes in electrolyte balance, PRA, P-A II, P-F, and P-aldo occurred in 3 subjects receiving 'sham' infusions. Additional experiments in subjects treated with propranolol or indomethacin allowed the conclusion that the effect of ACTH on PRA and P-A II is not mediated by renal beta-adrenergic receptors, but perhaps (partially?) by prostaglandins. Since the infusion rate of ACTH was not much higher than the secretion rate of ACTH in the early morning hours, it is possible that ACTH is physiologically involved in the regulation of renin secretion. PMID- 6289586 TI - ACTH suppression after oral administration of cortisone in Addisonian and adrenalectomized patients. AB - Plasma cortisol and the corresponding ACTH concentrations were determined before, and for 6 h following a single oral dose of 25 mg cortisone acetate in 7 patients with Addison's disease, 6 patients adrenalectomized for Cushing's disease and 1 patient adrenalectomized for congenital adrenal hyperplasia. The basal plasma cortisol concentrations 12 h after an evening dose of cortisone acetate 12.5 mg were below 100 nmol/l, and the corresponding ACTH concentrations were markedly elevated in all patients. Great interindividual variations were found in cortisol peak concentrations (Cmax) and the time to peak values, but without significant differences between the two patient groups. The maximal ACTH suppression occurred within 60-330 min after the cortisol Cmax, and was not significantly different in the two groups. The suppressed plasma ACTH concentrations were considerably above normal in 3 of the patients with Addison's disease and in 4 of the 6 patients adrenalectomized for Cushing's disease, including 2 patients with Nelson's syndrome. A similar degree of impaired ACTH suppression in patients with Addison's disease as in adrenalectomized patients suggests the occurrence of a secondary hypothalamic-pituitary dysfunction with ACTH hypersecretion in Addison's disease. The adequacy of the commonly used adrenocortical replacement therapy and its possible relation to the impaired ACTH suppression is discussed. PMID- 6289587 TI - Conversion of 22S-hydroxy-cholesterol and its effects on the metabolism of other sterols in rat adrenal cells and bovine adrenal mitochondria. AB - This study provides evidence that 22S-OH-cholesterol inhibits the conversion of 25-OH-cholesterol but has no effect on the conversion of 22R-OH-cholesterol. The latter sterol is an intermediate in the cholesterol side-chain cleavage, whereas for the conversion of 25-OH-cholesterol into pregnenolone the complete side-chain cleaving enzyme system is necessary. This complements a previous study in which it was shown, that 22S-OH-cholesterol has an inhibitory effect on the ACTH induced conversion of cholesterol into corticosterone in isolated rat adrenal cells. The available evidence thus suggests an inhibition by 22S-OH-cholesterol of the first step in the cholesterol side-chain cleavage. The results, obtained from the experiments with rat adrenal cells and with bovine adrenal mitochondria, allow the hypothesis, that a causal relationship exists between conversion of 22S OH-cholesterol and production of corticosterone, respectively pregnenolone. We conclude, that 22S-OH-cholesterol is a substrate for steroid production in the adrenal cell. This sterol inhibits the ACTH-stimulated corticosterone production. The site of this inhibition is located at one of the first steps in the cholesterol side-chain cleavage, probably the binding of cholesterol to the cytochrome P450-complex. PMID- 6289588 TI - Catecholamine stimulation of cyclic AMP and progesterone production in rat corpora lutea of different ages. AB - In vitro effects of catecholamines (adrenaline and noradrenaline) and adrenergic antagonists on adenosine 3',5'-cyclic monophosphate (cAMP) and progesterone production by rat corpora lutea (CL) of different ages (1-8 days old) were studied. To obtain defined ages of CL a pregnant mare's serum gonadotrophin (PMSG) model was used. The effect of catecholamines on cAMP decreased with luteal age while the effect on progesterone production was maximal on 5 day old CL. The beta-blocker propranolol inhibited the effects of catecholamines in concentrations around 10-(5) M. The effects of LH could only be inhibited with higher doses of propranolol known to exert unspecific effects. These results support the theory that LH and catecholamine effects on rat corpora lutea are mediated through different receptors. PMID- 6289589 TI - Pituitary and testicular functions in sexually mature rhesus monkeys under high dose LRH-agonist treatment. AB - Treatment with high doses of LRH-agonists leads to a down-regulation of testicular LH/hCG receptors and is accompanied by a suppression of spermatogenesis in some laboratory animals. In order to test whether this may provide an approach to male fertility control, 4 adult rhesus monkeys were treated with very high doses of LRH-agonist, D-Ser(TBu)6-LHR-ethylamide (100 micrograms daily) for 10 weeks during the breeding season. There was a decrease in testicular volume after 3 weeks of treatment. Serum LH and testosterone levels were suppressed. Sperm counts were stimulated during the first 6 weeks of treatment. At the end of treatment sperm counts, although lower, were still in the normal range. The chronic treatment schedule resulted in a decrease in pituitary responsiveness to an acute challenge with 4 micrograms agonist iv, indicating a desensitization of the pituitary. However, the testosterone response remained unchanged. This study using extremely high doses of the agonist indicates that, compared to the rat, male primates are far more resistant to the suppressive effects of LRH-agonists. PMID- 6289590 TI - Immunocytochemical study of the hypophysis in 25 dogs with pituitary-dependent hyperadrenocorticism. AB - Pituitary adenomas were found in 21 (84%) of 25 dogs with spontaneous pituitary dependent hyperadrenocorticism. Six dogs had pars intermedia adenomas, whereas 15 had tumours of the pars distalis. Diffuse corticotroph cell hyperplasia was found in 1 of the 4 pituitaries without adenoma; in 2 dogs with pituitary adenoma, coexisting hyperplasia of the surrounding corticotrophs was also present. Immunocytochemical staining of the pituitaries revealed positive staining for ACTH, beta-lipotrophin, and beta-endorphin in the majority of both pars distalis and pars intermedia adenomas. The most frequent and intense staining was found with anti-beta-endorphin. In most part intermedia tumours, many cells stained strongly for alpha-MSH; double immunostaining of one pars intermedia adenoma for ACTH and alpha-MSH showed that some tumour cells stained only for ACTH or alpha MSH whereas others contained both peptides. Only occasional cells stained for alpha-MSH in pars distalis adenomas. PMID- 6289591 TI - The discrepancy between plasma 11-deoxycortisol and ACTH concentrations in a single dose metyrapone test in normal men. AB - A single dose methyrapone test (MTP test) was carried out on 6 normal men by administering 1.0 g of metyrapone at 08.00-09.00 h with and without dexamethasone (DXM-MTP test) pre-treatment. Plasma 11-deoxycortisol, pregnenolone, ACTH and cortisol were measured before administration of the drug, and at hourly intervals for 6 h. In the MTP test, 11-deoxycortisol increased significantly at 1 h with a peak at 5 h, whereas significant increases in pregnenolone and ACTH were not seen until 3 h. There was a definite decrease in the cortisol level at 1 h with the lowest level measured at 2 h. Thus, a time discrepancy between plasma 11 deoxycortisol and ACTH concentrations was observed. The increase in 11 deoxycortisol after metyrapone should be divided into two phases: the increase in phase II (after 3 h) is due to the pituitary ACTH reserve, and that in phase I (the first 2 h) is due to some mechanism other than the pituitary ACTH reserve. The increased amount of 11-dexoycortisol in phase II (218.1 nmol) occupied 60.5% of the total increased amount in phases I and II (360.6 nmol). The cortisol/(cortisol + 11-deoxycortisol) ratio reached its lowest point 3 h after metyrapone treatment. This might be due to the initiation of an additional surge in 11-deoxycortisol by the ACTH reserve at 3 h. PMID- 6289592 TI - Simultaneous measurement of beta-endorphin, lipotrophins and met-enkephalin in phaeochromocytomas. AB - Tissue concentrations of immunoreactive lipotrophin, beta-endorphin, and met enkephalin were determined in 10 phaeochromocytomas, 3 of which were responsible for the ectopic ACTH syndrome. Lipotrophin and beta-endorphin immunoreactivities could be detected in all cases, whether or not Cushing's syndrome was present, and their tissue concentrations were significantly correlated (r = 0.95, P less than 0.001). Chromatographic studies showed that gamma-lipotrophin and beta endorphin were the main peptides in the tumours. Met-enkephalin immunoreactivity was also found in all tumours examined, at much higher concentration and showing no correlation with either lipotrophin or beta-endorphin immunoreactivity. Although beta-endorphin and met-enkephalin are thought to originate from different precursor molecules, these data show that the two opioid peptides may be secreted by the same tumour. The evidence for excess secretion of opioid peptides and their pathological significance in phaeochromocytomas remain to be established. PMID- 6289593 TI - A comparative study of complex mitochondrial DNA in human lymphocytes transformed by Epstein-Barr virus and PHA. AB - Mitochondrial DNA (mt DNA) molecules, isolated from normal human lymphocytes, lymphocytes stimulated with phytohemagglutinin (72 h) and a cultured Burkitt's lymphoma cell line, were examined by electron microscopy. Only monomeric forms of mt DNA were found in normal human lymphocytes, whereas in the other cell types, catenated forms were also observed. Unicircular complex forms were apparently absent in all the cell types studied, suggesting that these forms are not a common malignant transformation in human cells. No clear correlation between mitochondrial modifications--as detected by stereological analysis--and the presence of catenated forms could be established. It is suggested that other mechanisms rather than replication of mt DNA would appear to be the mechanism responsible for the formation of these forms. PMID- 6289595 TI - Rosenthal fibers in glioblastoma multiforme. PMID- 6289594 TI - Measurement of edema in the nervous system. Use of Percoll density gradients for determination of specific gravity in cerebral cortex and white matter under normal conditions and in experimental cytotoxic brain edema. AB - A method is presented by which density measurements can be performed on samples from cerebral cortex and white matter of normal and intoxicated animals using nontoxic ingredients as an alternative to the bromobenzene-kerosene technique described by Nelson et al. (1971). A continuous density gradient is prepared in a calibrated glass cylinder by using a new product, Percoll, which consists of colloidal silica particles coated with polyvinyl pyrrolidone. The gradient is stable and the same column can be used for repeated experiments over a long period of time. Interactions between the gradient media and the samples are evaluated and various methodological aspects concerning removal and handling of the tissue samples are presented. Experiments with acute triethyltin (TET) intoxication in the mouse and the hamster show that the Percoll technique can be used as an alternative to the bromobenzene-kerosene method in quantitative studies on cytotoxic brain edema. PMID- 6289596 TI - Intra-astrocytic glycogen granules and corpora amylacea stain positively for polyglucosans: a cytochemical contribution on the fine structural polymorphism of particulate polysaccharides. AB - A cytochemical procedure for polysaccharides was carried out on a brain biopsy specimen, the thin-section study of which had shown excess glycogen granules and the corpora amylacea variety of polyglucosan bodies. Both granules and amyloid bodies were stained positively in contrast to the remaining structures of the brain tissue which remained unstained. This demonstrates that beta-granules as well as filamentous and amorphous components of amyloid bodies are just different aspects of the polysaccharide molecule. Up to now the same kind of cytochemical evidence has been supplied for Lafora bodies of human material and Lafora-like bodies of rat material. The present study on corpora amylacea of human material shows that amyloid, Lafora, and Lafora-like bodies all behave the same way when stained for polysaccharides. PMID- 6289597 TI - Is the relaxing effect of beta-adrenergic agonists on the human myometrium only transitory? AB - The beta-adrenergic receptor function in myometrial strips taken from pregnant women at cesarean section was studied. The material consisted of myometrial strips from 10 patients treated with terbutaline for threatening premature parturition and an untreated control group of 10 patients. The basal cAMP levels were lower and the cAMP production after receptor stimulation in vitro was less in myometrial strips taken from terbutaline-treated women than in the untreated control group. The phosphodiesterase activity was greater in myometrial strips taken from terbutaline-treated women, indicating a faster rate of degradation of cAMP in these women. Studies on the beta-adrenergic receptor function in leukocytes from venous blood samples during treatment with terbutaline for preterm labor demonstrated a gradual decline in receptor function, as shown by a successive reduction in cAMP production after stimulation with a beta-adrenergic agonist in vitro. The results presented may explain the merely temporary inhibitory effect on uterine contractions often obtained during treatment with beta-mimetics. It is suggested that intermittent usage or combined treatment with drugs inhibiting phosphodiesterase activity might improve the results in the treatment and prevention of premature birth. PMID- 6289598 TI - Development of tolerance to the metabolic actions of beta 2-adrenoceptor stimulating drugs. PMID- 6289599 TI - Influence of argon laser stapedotomy on cochlear potentials. I: Alteration of cochlear microphonics (CM). AB - In guinea pigs the cochlea microphonics (CM) were recorded during and after argon laser stapedotomy. Short termed drastic depression of CM was observed. This effect involved the entire frequency range of the cochlea. The recovery was mostly complete and often started already during laser irradiation or at the end of the laser exposure. A marked sensitivity loss of the cochlea only occurred with large stapedotomy holes followed by massive perilymph leakage. A momentary heat related instability of the entire stapes is suggested to cause the CM depression. In addition we observed a thermoacoustic effect on the CM. This laser evoked response seems to cause no harm to the cochlea. PMID- 6289600 TI - Non-dysplastic villous changes in endoscopic biopsies in ulcerative colitis with carcinoma. AB - Three cases of long-standing ulcerative colitis and invasive adenocarcinoma are described. In all three cases, pre-operative rectal and/or colonoscopic biopsies demonstrated the presence of a villous adenoma: two without dysplasia and the third with moderate dysplasia. The presence of structures compatible with villous adenoma should be, even in the absence of epithelial dysplasia, an indication for panproctocolectomy in patients with long-standing ulcerative colitis. PMID- 6289601 TI - Sphingomyelinase activity of Staphylococcus aureus strains from recurrent furunculosis and other infections. AB - The sphingomyelinase (beta-hemolysin) production by 180 Staphylococcus aureus strains was determined in a qualitative enzyme impression test (EIT) and all but 4 strains in a quantitative enzyme assay test (EAT) using 14C-sphingomyelin as test substrates. A total of 59 strains (33%) were positive for enzyme production in the EIT. The highest frequency was found in strains isolated from recurrent furunculosis (45%) as compared with strains from chronic osteomyelitis, tropical pyomyositis and healthy nasal carriers (18, 8 and 19%, respectively). High producers of sphingomyelinase in EAT were mostly found among furunculosis strains, phage type 55 and/or 71. The furunculosis strains lysed by phages 3A and/or 3C mostly had a low sphingomyelinase production. In 3 families, high- and low- producing strains of the same phage group were isolated from 2 different members of the same family. The high-producing strains caused more intense skin lesions. PMID- 6289602 TI - Methane and hydrogen production by human intestinal anaerobic bacteria. AB - The gas above liquid cultures of a variety of human intestinal anaerobic bacteria was sampled and analysed by headspace gas chromatography. Hydrogen production was greatest with strains of the genus Clostridium, intermediate with anaerobic cocci and least with Bacteroides sp. Very few strains produced methane although small amounts were detected with one strain of B. thetaiotaomicron, C. perfringens and C. histolyticum. There may be a relationship between these anaerobic bacteria and several gastrointestinal disorders in which there is a build up of hydrogen or methane in the intestines. PMID- 6289603 TI - The influence of age and sex on weight variation in rats treated chronically with lithium chloride. AB - Weight increase is a common side-effect after chronic lithium salt treatment in man. This effect has not always been found in animal models using the rat. The possibility that the age and sex of animals might interfere with the results was studied, using groups of male and female rats, 2, 5, and 12 months old, injected intraperitoneally twice daily for 28 days with a solution of lithium chloride, or sodium chloride as control. The dose used was 1.5 meq./kg body weight (total daily dose of 3.0 meq./kg). In 2 month old animals, lithium induced an increase in weight significantly greater than that of the controls in the female group, while the weight of the male rats in this age group remained indistinguishable from the control rats. The 5 and 12 month old lithium treated female rats showed no differentiation in weight. Lithium treatment of the males in these two age groups caused a large number of deaths and, considering only those animals which survived, average weights were significantly less than those of control in the 5 month olds rats, and equal to control in the 12 month olds. It is concluded that age and sex have to be taken into consideration when studying the effects of chronic lithium treatment on the weight of rats. PMID- 6289604 TI - Adenylate cyclase in rat synaptosomal plasma membranes and caudate nucleus homogenate: effects of dopamine, E prostaglandins, guanyl-5'-yl-imidodiphosphate and morphine. AB - The adenylate cyclase in two particulate preparations from rat brain, a homogenate from caudate nucleus (CN-homogenate) and a synaptosomal plasma membrane fraction (SPM-fraction) from whole rat brain was investigated. Stimulation of the enzyme by dopamine and prostaglandins E1 and E2 was found in the CN-homogenate while only a weak prostaglandin E1 and E2 stimulation and no dopamine stimulation could be found in the SPM-fraction. Guanyl-5'-yl imidophosphate (GppNHp) and NaF could stimulate the adenylate cyclase in both preparations. Morphine up to 10(-5) M altered neither the basal enzyme activity nor any of the stimulated enzyme activities. PMID- 6289605 TI - Assay of adenylate cyclase in rat brain homogenates and lysed turkey erythrocytes: two different methods give different results with brain but not with erythrocytes. AB - Experiments showed that the cyclic AMP (cAMP) fraction isolated by alumina or Dowex 50/alumina chromatography from brain adenylate cyclase reaction mixtures contained 32P radioactivity 10-12 times in excess of that which could be accounted for by determination of cAMP using binding assays. No such discrepancy was found when lysed turkey erythrocytes were assayed. This indicated that special precautions must be taken for the purification of 32PcAMP from brain adenylate cyclase assays due to the formation of 32P-labelled contaminants. PMID- 6289606 TI - Effects of methyl mercury and triethyllead on Na+K+ATPase and pyruvate dehydrogenase activities in glioma C6 cells. AB - The effect of methyl mercury (MeHg) and triethyllead (Et3Pb) on the membrane bound SH-enzymes Na+K+ATPase and pyruvate dehydrogenase (PDH) was studied in relation to the effect on the galactosyl ceramide sulfotransferase (CST) and to morphological changes in glioma C6 cells. Two-day-old cultures were incubated for 1 or 20 hrs with 5-30 microgram MeHgC1 and 2-8 microgram Et3PbC1/mg cell protein. The results show that both compounds induced morphological changes and a reduction of CST activity at growth inhibitory concentrations. A less marked reduction of Na+K+ATPase was induced with increasing exposure time only in MeHgC1 treated cultures, and PDH activity was not affected by either of the compounds under the experimental conditions. Thus, an interference with Na+K+ATPase and PDH activities do not appear to be a primary effect of MeHg and Et3Pb intoxication. PMID- 6289607 TI - Adrenergic transmission and prostaglandins. PMID- 6289608 TI - Constellation system of transmitter receptors at the terminals of the sensitive nerves of eye cornea, arterial blood vessels, heart and uterus. PMID- 6289609 TI - Herpes simplex virus (HSV) antibodies in child psychiatric patients and normal children. AB - The prevalence of herpes simplex virus (HSV) antibodies has been investigated in 123 child psychiatric patients and 86 normal children. HSV antibodies were measured by ELISA technique. The prevalence of HSV antibodies in different diagnostic groups (conduct disorder, emotional disorder, hyperkinetic syndrome, anorexia nervosa, infantile autism and borderline schizophrenia in childhood) was compared with age-matched normal children, but no significant differences were found. PMID- 6289610 TI - Radiation retinopathy in diabetes mellitus: report of a case. AB - A case of radiation retinopathy in a diabetic individual who received a total dose of 45 Gy for lymphoblastic lymphoma of the orbit is reported. The relationship between radiation retinopathy and diabetes mellitus is discussed. PMID- 6289611 TI - Resistance to prolonged irradiation in mice influenced by thyroid hormones and chemical radiation protectors. AB - After the end of 10 and 21 days' treatment of mice with dried thyroid gland (0.6 g/100 g diet) the effect of some radiation protectors against prolonged irradiation (38 mGy/min) increased. The combination of propranolol, phenobarbital and cystamine was more effective than a mixture of cystamine and 5 methoxytryptamine both in mice fed on a normal and a thyroid gland diet. A reduced oxygen consumption after the application of protectors reflected the degree of protection. The number of thrombocytes in peripheral blood increased after the end of thyroid treatment, which indicated a possible shift in the differentiation of stem cells in favour of megakaryocytes. PMID- 6289612 TI - Neoplasms in ovaries of CBA mice 90Sr-treated as foetuses. AB - Female CBA mice were 90Sr-contaminated in utero by amounts ranging from 46 to 740 kBq given to the pregnant dams on the 19th day post coitum. The females were killed at an average age of 10 months. The ovaries were microscopically analysed and morphologic changes were recorded. Multiple corpora lutea were frequent in all but the two highest dose groups. Ovarian cysts were frequent as well. Proliferative events such as hyperplasia of luteinized interstitial cells and down-growths of the germinal epithelium did not occur until a dose of 185 kBq. The findings indicate a non-linear dose-tumor relationship, but also a direct proportion of animals with down-growths and those with tubular adenomas beyond a threshold of 14 per cent. PMID- 6289613 TI - Influence of sera from tumor bearing mice on immune reaction. AB - Immunopotentiation abilities of sera from mice bearing fibrosarcoma or lymphoma for various periods of time were assayed using the plaque forming cell (PFC) technique. The influence of such sera on cell proliferation, evaluated by the spleen index, was determined in lethally irradiated mice reconstituted with syngeneic bone marrow cells. Sera from tumorous animals stimulated the number of PFC significantly, fibrosarcoma sera being more effective in the advanced stage and lymphoma in the initial phase. Sera from mice with both tumors in the early phase increased spleen indices significantly, and sera from mice with advanced tumors were less effective. The possible role of lymphokines in these events is discussed. PMID- 6289615 TI - Ferrous sulphate dosimeter for control of ionization chamber dosimetry of electron and 60Co gamma beams. AB - A check of the dosimetry using ionization chambers and applying the procedures described in the Nordic dosimetry protocols (NACP 1980, 1981) was carried out using the ferrous sulphate dosimeter as reference. Measurements have been carried out in electron beams in the energy range 2.8 to 27.5 MeV and in 60Co gamma beams. Consistent results were obtained with the two systems within about one per cent. Larger differences were only observed in electron beams containing a large proportion of scattered low energy electrons and contaminating roentgen rays which were not corrected for in the ionization chamber method. PMID- 6289614 TI - Radiation-dose dependence of the formation of micronuclei in misonidazole treated cell cultures. AB - The effect of misonidazole at different concentrations on the anoxic radiation sensitivity of Chinese hamster cells was investigated using the frequency of radiation induced micronuclei as criterion. The result indicates that, in a high radiation dose region, sensitization with a dose modifying factor of about 1.9 and 1.3 occurs after treatment with the substance at a concentration of 8 and 0.2 mmol/l, respectively. In a low dose region the corresponding values were 1.7 and 0.8. It was concluded that high concentration of the substance in combination with high radiation doses are most beneficial. PMID- 6289616 TI - Invasive squamous cell carcinoma of the uterine cervix. V. Late local recurrences after radiation therapy. AB - In 26 patients with late local recurrent invasive squamous cell carcinoma of the uterine cervix, primary and recurrence biopsies have been compared with respect to a malignancy grading system. In patients with 16 points or more (n = 13) there was no significant change between estimations. In patients scoring less then 16 points (n = 13), 7 patients had a significant rise in tumour host score indicating the possibility of new prognostic factors occurring. The concept of recurrence is discussed against the background of these findings. PMID- 6289617 TI - Carcinoma of the nasopharynx: results of radiation therapy. AB - Of a series of 186 patients with nasopharyngeal neoplasm 138 cases were treated with curative irradiation between 1959 and 1978. All cases were reviewed and retrospectively classified according to the TNM system. The 5-year crude survival rate was 39.5 per cent. A better prognosis was found for women and for younger age groups. The degree of neck node involvement had more prognostic value than the extent of the primary tumor. The causes of failure were analyzed. In 35 cases distant metastases were observed. PMID- 6289618 TI - Radical irradiation in the primary management of localized breast carcinoma. AB - One hundred-twelve females with stage I-III breast carcinoma were treated primarily with radical megavoltage irradiation. The loco-regional area received 50 Gy/5 weeks with a boost to the primary site of 10 Gy/1 week to 20 Gy/2 weeks. Local control rates were as follows: 94 per cent in stage I, 86 per cent in stage II and 69 per cent in stage III. Survival rates were comparable to those reported by other authors. Complications correlated with disease stage and extent of radiation fields. PMID- 6289619 TI - Tissue culture of primary rat anterior pituitary cells. PMID- 6289620 TI - Interplay between opioid peptides and pituitary hormones. PMID- 6289621 TI - Forms of opioid peptides circulating in human blood: investigation with multiple radioimmunoassays and radioreceptor assay. PMID- 6289622 TI - Plasma beta-endorphin and met-enkephalin in physiological and pathological conditions. PMID- 6289624 TI - In vitro studies of the biosynthesis and release of alpha-N-acetyl-beta-endorphin from the intermediate lobe of rat pituitary. PMID- 6289623 TI - Gene expression of peptides: the pro-opiomelanocortin model. PMID- 6289625 TI - Hydrolysis of enkephalins by human converting enzyme and localization of the enzyme in neuronal components of the brain. AB - Homogeneous human CE hydrolyzed Met5-enkephalin fastest of the biologically active substrates tested. The inactivation of Met5-enkephalin was inhibited by antiserum to Ce and by specific inhibitor of CE (MK 421; I50 = 8 x 10(-9)M). Direct radioimmunoassay of CE extracted from basal ganglia indicated immunological identity of lung, kidney, and brain CE. Immunocytochemically, CE was demonstrated in the choroid plexus, where the enzyme appears to be bound, as in kidney and gut, to the surface of the epithelial cell bathed by luminal fluid. CE immunoreactivity in brain parenchyma was demonstrated in neuronal elements in the hippocampus, hypothalamus, neocortex, globus pallidus, and substantia nigra. PMID- 6289626 TI - Receptors and post-receptors alterations induced by opiates and solubilization of opiate receptors. PMID- 6289627 TI - Affinity labeling and purification of the opiate receptor from rat brain. AB - Affinity labeling of the opiate receptor has been performed on neural membranes from rat brain utilizing[125I]-14-bromoacetamidomorphine, an opiate agonist, and [125I]-14-chloracetylmorphine, an antagonist. With the use of SDS gel electrophoresis it could be shown that the agonist labeled three proteins with molecular weights of 43,000, 35,000 and 23,000, whereas the antagonist only labeled the 23,000 component. The preferential labeling of the 23,000 protein by the antagonist suggests that this component may be a primary recognition site for opiate antagonists. Calcium was stimulatory to the affinity labeling of all three proteins while sodium was inhibitory. With the use of affinity columns prepared by conjugating either ligand to omega-aminohexyl Sepharose, a receptor complex was obtained consisting all three proteins. Stereospecific opiate binding was demonstrable in the complex prepared from either column. PMID- 6289628 TI - Recent work on opiate receptors: heterogeneity and solubilization. PMID- 6289629 TI - Rational design and biological properties of highly specific mu and delta opioid peptides. PMID- 6289631 TI - Met-enkephalin and related peptides in man. PMID- 6289630 TI - Enkephalin and beta-endorphin as mediators of electro-acupuncture analgesia in rabbits: an antiserum microinjection study. PMID- 6289632 TI - Putative new regulatory peptides in mammals: phenypressin (Phe2-Arg8-vasopressin) and mesotocin (Ile8-oxytocin). PMID- 6289634 TI - Beta-Endorphin receptor and its possible relationship to other opioid receptors. PMID- 6289633 TI - Characterization of multiple forms of beta-E in pituitary and brain: effect of stress. PMID- 6289635 TI - Characterization of synaptic phosphoproteins modulated by ACTH. PMID- 6289636 TI - Polyamines in mammalian tumors. Part II. PMID- 6289637 TI - Epstein-Barr virus antigens-a challenge to modern biochemistry. PMID- 6289638 TI - Kinetic studies of cytochrome-c-oxidase: significance of different functional states of the enzyme. PMID- 6289639 TI - Non-heme components of electron transfer systems: interactions and conformational effects. PMID- 6289640 TI - Intermediates in the reduction of dioxygen y laccase and cytochrome c oxidase. PMID- 6289641 TI - Structure and function of the redox site of cytochrome oxidase. PMID- 6289642 TI - Inactivation of microbial pyridoxal kinase by pyridoxal. AB - Pyridoxal kinase from Escherichia coli and bakers' yeast was inactivated by pyridoxal while the enzyme from rat and pig brain was not. The inactivation of the enzyme purified from E. coli was reversible and was rendered irreversible by the reduction with NaBH4. This finding as well as a similar inactivation by 5' deoxypyridoxal but not by 4'-deoxypyridoxine suggested that the inactivation was due to Schiff base formation. The suggestion was confirmed by the incorporation of tritium label into the enzyme by the reaction of the enzyme with [3H] pyridoxal followed by the treatment with NaBH4. Correlation between the loss of enzyme activity and the amount of pyridoxal bound to the enzyme showed that binding of pyridoxal to one crucial site completely inactivated the enzyme. Pyridoxine and 4'-deoxypyridoxine did not have a protective effect against inactivation indicates that the binding site was not the substrate site. The results of kinetic and equilibrium analyses were consistent with a one-step inactivation mechanism. PMID- 6289643 TI - Vitamin D and its metabolites. Advances in the diagnosis and treatment of rickets. AB - Diagnostic and therapeutical uses of vitamin D3 and its metabolites are reviewed. Special emphasis is dedicated to the fetomaternal relationships of 1,25 (OH)2 D3 and 25-OH-D3 at term. The serum levels of 1,25 (OH)2 D3 have been found to be higher in the maternal serum then in the corresponding fetus (85.3 pg/ml and 50.9 pg/ml, respectively). The highest serum levels of 1,25 (OH)2 D3 were found in October and the lowest ones in January showing that there is a dependence on the ultraviolet light. It has been found that there is a correlation between the fetomaternal serum levels of 1,25 (OH)2 D3 and 25-OHD. However, there is no correlation between the serum levels of 1,25 (OH)2 D3 and 25-OHD3, neither in the fetus nor in the mother. PMID- 6289644 TI - Health and the environment-a comparative study of agricultural and industrial workers in Nigeria. AB - Although several attempts have been made in the developed countries to study the effect of various environmental factors on morbidity, it is disappointing that in developing countries where very poor conditions exist, very little information is available on the subject. This study was therefore designed to examine the influence of the living and working environment on the state of health of two groups of Nigerian workers namely farmers and industrial workers. The study was carried out in two villages Badeku and Ewekoro. 200 farmers from Badeku and 150 industrial workers from Ewekoro were selected by appropriate sampling methods. Most of the data were obtained by means of a questionnaire administered by the authors. In addition to a detailed clinical examination of each subject, blood stool and urine specimens were obtained for laboratory investigations. A high incidence of hookworm and ascaris infection, anaemia, leg ulcer, malaria parasitaemia and onchodermatitis was found among the farmers. The industrial workers on the other hand had a higher incidence of chronic bronchitis and hypertension. Some of these findings have been attributed to poor environmental hygiene of the homes and work places, atmospheric pollution and other health hazards associated with living and working in the rural areas. PMID- 6289645 TI - Alpha 1-antitrypsin phenotypes in a Nigerian population. AB - Alpha 1-antitrypsin phenotypes of 350 normal Nigerian blood donors were determined by isoelectric focusing in polyacrylamide gel. The distributions obtained (MM, 66.86%; MS, 16.29%; MZ, 0.57%; SS, 2.0%; SZ, 0.28%) were different from those reported for caucasian populations. Forty-nine of these donors could not be classified into any of the above common phenotypes suggesting that some rare alleles occur in Nigeria. The probable cause and implications of the high frequency of the S gene in the Nigerian population are discussed. PMID- 6289647 TI - Gastric acid secretion using pentagastrin: dose-response studies. AB - The results of gastric stimulation using varying doses of pentagastrin in twenty five Nigerians (twenty cases of duodenal ulcer and five controls) are presented. Doses used were 4, 6, 8, and 10 micrograms/kg bodyweight. The optimal response was obtained using 6 micrograms/kg bodyweight in the test cases while the maximum acid output (MAO) was 15.14 mEq/hr (19 mEq/hr in males, 12.50 mEq/hr in females). The recommended dose for optimal response in Caucasians is 6 micrograms/kg bodyweight which correlates with the dose in our patients. However, the maximum acid output at this dose is higher in Caucasians. PMID- 6289646 TI - Metabolic consequences of mycoplasmal contamination of cell cultures. AB - KB cells originally derived from epitheloid carcinoma of the nasopharynx were found to be contaminated cells showed significantly higher glycolytic (FMP) and respiratory (TCA) rates when compared with non-contaminated or "cured" cells. The activity of the hexose monophosphate (HMP) shunt, an alternative pathway of glucose metabolism, was shown to be reasonably higher than normal KB cell levels. Treatment involving combination of heat (41 degrees C) and kanamycin (350 micrograms/cm3) for 21 hours was found to adequately and selectively inactivate the mycoplasma population. Following cure, the metabolism of the cell fell well within normal ranges. The treatment showed no deleterious effects of the KB cell population. The possibility and the significance of an independent hexose, monophosphate shunt activity in the mycoplasma population in addition to the already established partial TCA and EMP activities, and the overall significance of detection of carbohydrate metabolism in these organisms, are discussed. PMID- 6289649 TI - Anaesthetic aspects of the first successful open heart surgery at the University College Hospital, Ibadan, Nigeria. PMID- 6289648 TI - Early onset and significance of imcomplete left posterior hemiblock in Nigerians. AB - Following detailed clinical and electrocardiographic analysis, twenty-five patients with incomplete left posterior hemiblock without associated complete right or left bundle branch block are presented. First-degree atrioventricular block was relatively common in these patients and associated incomplete right or left bundle branch block was also present in a few. Hypertension was the commonest underlying cardiovascular disease but two of them, without any clinically overt cardiovascular disease, presented with a history of episodic dizziness or syncopal attacks. With the present knowledge of the anatomy and histopathology of intraventricular conduction system, the relevance of tendency for Africans to form excessive fibrous tissue has been emphasized. Considering the unexpectedly low average age of these patients, suggestions have been made with a view to reducing the rate of progression of such atrioventricular conduction defects to the need for implanted cardiac pacemakers. PMID- 6289650 TI - Investigation of the presence of Salmonella in two Nigerian meat packing plants. AB - Two large and very well established meat processing plants supervised by the Veterinary Public Health Department were examined to assess the incidence of Salmonella within the premises and on the processed carcasses. The overall incidence was 65 out of the 252 samples or 25.9%. Salmonella was found in forty one of 167 samples (25%) in one and twenty-four out of eighty-five samples (29%) in the other. The high incidence shows a high degree of probability of contamination of the products from both plants. The public health significance of the findings is emphasized and control problems are high-lighted. PMID- 6289652 TI - Torsion of testis in Nigerians. AB - Torsion in a black population is presented. The clinical features are the same as the white counterpart, but the mean age is, however, much higher. There is no bias against any side of testis. Physicians are to blame for the poor salvage rate. Gangrenous testes should not be conserved and early diagnosis and treatment remain the only means of saving torted testis. Prophylactic fixation of contralateral testis is mandatory and must not be postponed. External manual detorsion is not a substitute for surgery and should not remove the sense of emergency. Relevant pathology, the mechanism of torsion and differential diagnosis are discussed. PMID- 6289651 TI - Curing antibiotic resistance in Bacteroides species by aminoacridines and ethidium bromide. AB - Three clinical isolates of Bacteroides fragilis and one faecal isolate of B. thetaiotaomicron resistant to one or more of the antibiotics chloramphenicol, erythromycin, clindamycin and tetracycline, were cured of their resistance markers by treatment with subinhibitory levels (16 micrograms/ml) of acriflavine, acridine orange and ethidium bromide. Chloramphenicol, erythromycin and clindamycin resistance markers were cured en bloc after exposure to the agents for 24 hr but elimination to tetracycline resistance markers required longer incubation (17-21 days) with the reagents. The minimum inhibitory concentrations of the antibiotics for the Bacteroides strains before and after elimination are compared. Elimination of these antibiotic resistances indicates that the resistance markers are located as extrachromosomal plasmids (R-factors). The emergence of plasmid-mediated antibiotic resistances may compromise treatment of bacteroides infections and may also provide a reservoir of antibiotic resistance in the intestinal flora. PMID- 6289653 TI - Acquired immunodeficiency in homosexual men. PMID- 6289654 TI - Effects of angiographic contrast medium on isolated canine coronary arteries. AB - In isolated canine coronary arteries previously contracted by high potassium concentration, angiographic contrast medium decreased active tension by 61 +/- 2%. The relaxant effect was dose dependent and was not prevented by beta-blockade with d-l-propranolol (10(-5) M). This effect was similar to that obtained with nitroglycerin (10(-6) M), and further relaxation was evident when this vasodilator was administered after exposure to the contrast medium. When arteries were precontracted by alpha-receptor stimulation with norepinephrine (10(-5) M) at normal potassium concentration, a maximal relaxation of 83 +/- 6% was elicited after exposure to contrast medium. The relaxant effect could not be reproduced by a similar increase in osmolarity brought about by addition of sucrose. When arterial strips were processed by radioimmunoassay for dosage of cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP) after the relaxing action of the contrast medium occurred, a decrease in cAMP from 2.61 +/- 0.86 to 0.63 +/- 0.1 pmol/mg protein (p less than 0.05) was observed, whereas no significant changes in cGMP were detected. These nucleotides do not appear to be involved in the relaxant effect of the dye in the same way as they are when relaxation is elicited by some other coronary vasodilators. PMID- 6289655 TI - Dietary intakes and stool characteristics of patients with the irritable bowel syndrome. AB - Detailed dietary records were obtained from 30 women with the irritable bowel syndrome and 25 healthy, asymptomatic women. Stool transit times and wet stool weights were measured. Analysis of the dietary data revealed no significant difference between energy, protein, fat, carbohydrate, or total fiber intakes. Total dietary fiber intake was 17 g/day in the irritable bowel group and 18 g/day in the control group, but vegetable fiber intake was significantly lower in the patients (6.6 g/day) than the controls (8.2 g/day). Cereal and fruit fiber were comparable for both groups as were the stool transit times and wet stool weights. These results do not support the hypothesis that symptoms of the irritable bowel syndrome are directly related to total or cereal fiber depletion. PMID- 6289656 TI - On deafness, cytomegalovirus, and neonatal screening. PMID- 6289657 TI - Symptomatic congenital cytomegalovirus. Disorders of language, learning, and hearing. AB - Seventeen patients with symptomatic congenital cytomegalovirus (CMV) were studied longitudinally, with emphasis given to disorders of language, learning, and hearing. At a mean age of 5.5 years (range, 1 go 10 years), nine children (53%) performed in the retarded range. Eleven (65%) experienced sensorineural hearing loss, in three of whom it was progressive. Developmental verbal dyspraxia was documented in two children and suspected in a third. Disabilities in several areas of the learning process exhibited by four children with normal intelligence and hearing loss. Although the effects of congenital CMV were diverse, all of the children had developmental disorders that necessitated special education. Such patients require longitudinal follow-up that includes more than tests of intelligence and hearing. All areas of development must be evaluated to appreciate the full effect of CMV encephalitis in utero. PMID- 6289659 TI - Electrophysiologic changes in workers with "low" blood lead levels. AB - In spite of numerous studies, the minimum level of lead exposure at which "sub clinical" electrophysiologic abnormalities appear is still under discussion. Furthermore, it has not been clarified whether the electrophysiologic changes are directly related to PbB levels or to duration of exposure. This study was conducted on a group of 62 subjects occupationally exposed to lead with average blood lead levels below 50 microgram/100 ml and durations of exposure of less than 10 years. A reduction of motor and sensory nerve conduction velocities and sensory action potential amplitude of the median nerve was found in the subjects exposed to lead, as compared with a control group. Such abnormalities were already present in workers with the lowest blood lead levels, but were more severe in workers whose blood lead levels had exceeded 70 microgram/100 ml, even if this occurred only once in the last two years. The electrophysiologic changes did not correlate with duration of exposure but occurred very soon after initial exposure to lead. PMID- 6289658 TI - Demonstration of the heterozygous state for I-cell disease and pseudo-Hurler polydystrophy by assay of N-acetylglucosaminylphosphotransferase in white blood cells and fibroblasts. AB - The biochemical abnormalities of I-cell disease (mucolipidosis II) and pseudo Hurler polydystrophy (mucolipidosis III) can be explained by a deficiency of the enzyme UDP-N-acetylglucosamine:lysosomal enzyme N-acetylglucosamine-1 phosphotransferase. We demonstrate here that obligate heterozygotes for these autosomal recessive diseases have intermediate levels of this enzymatic activity in homogenates of peripheral blood white cells and in extracts from cultured fibroblasts. This finding provides further evidence that the enzyme deficiency is the primary genetic defect in these diseases. In addition, the previous observation that obligate heterozygotes for mucolipidosis III have elevations of total serum beta-hexosaminidase outside the range of normal was confirmed. In studies of three pedigrees of patients with mucolipidosis III, these techniques were used to score individuals at risk for the carrier state. PMID- 6289660 TI - Hepatomas: hormone receptors and therapy. AB - Prompted by studies that suggested a causal relationship between sex steroids and human liver cancer, we assayed specimens of hepatoma from five patients and adjacent liver tissue from three of the same patients for estrogen receptors. After finding that the assay material contained specific cytosolic receptor proteins for estradiol, we treated a second group of five patients who had hepatomas with progestin. This therapy resulted in tumor regression in two of these patients. PMID- 6289661 TI - Randomized study of high-dose versus low-dose methotrexate in the treatment of extensive small cell lung cancer. AB - Forty patients with extensive small cell lung cancer randomly received either high-dose or low-dose methotrexate with leucovorin rescue in combination with cycles of cyclophosphamide, doxorubicin, and vincristine alternating with cycles of VP-16, vincristine, and hexamethylmelamine. Nineteen patients were treated with the high-dose methotrexate regimen, and 21 received the low-dose methotrexate treatment protocol; both treatment groups were similar in median age, performance status and spread of disease. Response rates (74 percent for high-dose therapy; 67 percent for low-dose therapy), median survival (nine months versus nine months), and overall survival were similar for the two treatment groups. Myelosuppression was equivalent in both treatment groups but moderate to severe mucositis developed more often when patients were treated with the high dose methotrexate regimen as compared with low-dose methotrexate therapy (p less than 0.001). Central nervous system recurrences developed after three patients received high-dose methotrexate therapy. This study indicates that when used with other antineoplastic agents, high-dose methotrexate therapy does not improve the remission rate or survival nor does it decrease central nervous system metastasis in patients with small cell lung cancer when compared with standard doses of methotrexate; high-dose methotrexate is associated with greater cost and toxicity. PMID- 6289662 TI - Primary empty sella syndrome, ACTH hypersecretion, and normal adrenocortical function. Report of two cases. AB - Although the primary empty sella syndrome (PESS) is associated with normal endocrine function or subtle pituitary insufficiency, pituitary hormone hypersecretion associated with PESS has also been recognized. ACTH hypersecretion and primary empty sella syndrome have previously been reported in patients with either Cushing's disease or Addison's disease. This report describes two unique patients with ACTH hypersecretion, primary empty sella syndrome, and normal cortisol dynamics. The investigators speculate that this association may have resulted from infarction of hyperplastic adenohypophyseal corticotrophes due to production of an ACTH peptide with reduced biologic activity. These two cases emphasize that primary empty sella syndrome may be associated with ACTH hypersecretion and normal adrenocortical function. PMID- 6289663 TI - Angiotensin-converting enzyme in diseases of the liver. AB - Serum angiotensin-converting enzyme activity was measured in various diseases of the liver. Activity increased in progressive order in patients with chronic persistent hepatitis, chronic aggressive hepatitis, and liver cirrhosis. Activity was increased also in patients with acute hepatitis. On the other hand, patients with fatty liver had normal angiotensin-converting enzyme activity and patients with extrahepatic obstructive jaundice showed subnormal activity. Although the mechanism for these enzymatic changes in diseases of the liver remains to be elucidated, serum angiotensin-converting enzyme determination may be useful in the diagnosis of diseases of the liver under certain conditions. PMID- 6289664 TI - Prolactin binding sites on human chorion-decidua tissue. AB - An effective procedure has been developed and utilized to demonstrate the presence of prolactin receptors on the plasma membranes of human chorion-decidua cells. Particulate fractions from human chorion-decidua sedimenting between 1,500 and 45,000 x g display optimal binding of 215I-labeled ovine prolactin when incubated at a membrane protein concentration of 200 micrograms per assay tube for 2 hours at 22 degrees C. Specific binding was increased by pretreatment of the membrane particles with 5M magnesium chloride to remove endogenous prolactin. These receptors show binding parameters (affinity, 0.92 x 10(9) L/mode; capacity, approximately 80 fmoles/mg) similar to those of lactogenic receptors in the rabbit mammary gland and, the rabbit and rat liver. The presence of prolactin receptors in human chorion-decidua suggests that may play a role in mediating local action(s) of prolactin such as involvement in the decidualization reaction or in maintaining fetal osmoregulation. PMID- 6289665 TI - The effect of prostaglandins on the multiplication and cell-to-cell spread of herpes simplex virus type 2 in vitro. AB - Herpes simplex virus type 2 (HSV-2) after infecting an individual has the ability to remain latent in nervous tissues. The factors that control herpesvirus infection, latency, and reactivation are poorly understood. Fever, menstruation, emotional stress, exposure to sunlight, and surgical resection have been associated with activation of latent herpes. The situations which activate latent herpes are associated with a local or systemic rise in prostaglandins. The data presented show that prostaglandin F2 alpha (PGF2 alpha) and E2 (PGE2) enhanced cell-to-cell spread of HSV-2 in an in vitro model. Ibuprofen, a prostaglandin inhibitor, suppressed HSV-2 multiplication as well as cell-to-cell spread. Prostaglandins may play an important role in herpesvirus infections and latency. PMID- 6289666 TI - Fatal gestational trophoblastic disease: an analysis of treatment failures. AB - Forty-eight of 399 patients referred to the John I. Brewer Trophoblastic Disease Center of Northwestern University Medical School from 1962 to 1979 for treatment of gestational trophoblastic disease (invasive mole or choriocarcinoma) died. All patients who died had histologically documented metastatic choriocarcinoma. The time from pregnancy event to treatment was greater than 4 months and/or the pretreatment human chorionic gonadotropin titer was greater than 100,000 IU/L in 64% of these patients. Seventy-one percent of fatal cases developed in association with term pregnancies, abortions, or ectopic pregnancies rather than hydatidiform moles. Fifty percent of patients who died had metastases to the liver, brain, and/or peritoneal cavity when they first presented for treatment. The most common causes of death were hemorrhage from one or more metastatic sites (42%) and pulmonary insufficiency (31%). Factors primarily responsible for the treatment failures in these patients were: (1) presence of extensive disease at the time of initial treatment; (2) inadequate initial treatment; and (3) failure or presently used chemotherapy protocols in advanced disease. Secondary chemotherapy, radiation therapy to sites other than the brain, and adjuvant surgical procedures failed to improve survival in these high-risk patients. PMID- 6289667 TI - Placental protein 5 in gestational trophoblastic disease: localization and circulating levels. AB - With the use of an indirect immunoperoxidase technique (peroxidase antiperoxidase) placental protein 5 (PP5) has been shown to be present in a high percentage of gestational trophoblastic tumors, especially the better differentiated ones. By sensitive radioimmunoassay, PP5 has further been found to be present in sera from patients with persistent gestational trophoblastic disease but not to the same frequency or extent as human chorionic gonadotropin. The only finding of clinical importance was the association in one case of high levels of PP5 and concurrent intravascular coagulation. PMID- 6289668 TI - Radioimmunoassay of placental protein 12: levels in amniotic fluid, cord blood, and serum of healthy adults, pregnant women, and patients with trophoblastic disease. AB - A radioimmunoassay for placental protein 12 (PP12) is described and the levels in amniotic fluid, cord blood, and serum of nonpregnant individuals, pregnant women, and patients with trophoblastic disease are presented. During pregnancy, the highest PP12 levels were found at 22 to 23 weeks (mean +/- SD, 169 +/- 123 ng/ml), and there was a transient decline at 32 to 33 weeks (63 +/- 23 ng/ml). In amniotic fluid, the levels were 100 to 1,000 times higher than in maternal serum. In cord blood at birth, the values were of the same magnitude as in maternal serum. Also healthy nonpregnant women and men had PP12-like immunoreactivity in serum. Nonpregnant women (9 to 47 ng/ml) had higher levels than men (undetectable to 21 ng/ml). Elevated levels up to 84 ng/ml were occasionally observed in trophoblastic disease, both hydatidiform mole and choriocarcinoma, but they bore no correlation with the human chorionic gonadotropin levels. On the basis of these results PP12 is not a suitable marker for trophoblastic disease. PP12 values in normal pregnancy provide the basis for the evaluation of PP12 levels in abnormal pregnancy. PMID- 6289669 TI - Rapid adrenocorticotropic hormone test. PMID- 6289671 TI - Animal model of human disease. Pleomorphic salivary adenoma. Virally induced pleomorphic salivary adenoma in the CFLP mouse. PMID- 6289670 TI - Immunohistochemical and immunoelectron-microscopic study of pituitary adenomas associated with Cushing's disease. A report of 13 cases. AB - Thirteen pituitary adenomas were removed from patients with Cushing's disease by the transphenoidal route. All cases demonstrated a typical histochemical and ultrastructural pattern. Immunocytochemical study by means of the immunoperoxidase technique and light or electron microscopy demonstrated 1-24/1 39 adrenocorticotropic hormone (ACTH) in all cases, lipotropin/melanotropin (beta LPH/beta-MSH) in 10 cases, beta-endorphin in 8 cases, and an absence of calcitonin in all cases. In addition, in 2 cases tumor tissue contained a few antiprolactin immunoreactive cells. These ACTH, beta-LPH, and beta-endorphin immunoreactivities may reflect either the peptides themselves or their precursors or intermediate products. The authors also suggest a possible intermediate-lobe like processing of beta-LPH leading to beta-endorphin production, which may act on PRL cells. In addition, no positive arguments for the existence of a common precursor for calcitonin and ACTH could be provided from this study. PMID- 6289672 TI - Hormone-induced cell death. Purification ad properties of thymocytes undergoing apoptosis after glucocorticoid treatment. AB - A high proportion of cortical thymocytes obtained from suckling rats undergo apoptosis on exposure in vitro to the glucocorticoid methylprednisolone. The apoptotic cells can be separated from apparently normal thymocytes by isopyknic centrifugation on Percoll gradients. This experimental system provides homogeneous populations of apoptotic cells and thus permits a more incisive study of this physiologic mode of cell death than has been hitherto possible. It is shown that apoptosis involves a sharp but transient increase in buoyant density, concomitant with the appearance of characteristic morphologic changes in nucleus and cytoplasm. More than 80% fo the chromatin of apoptotic cells has a molecular weight sufficiently low to resist sedimentation at 27,000g and consists of short oligonucleosome chains, apparently as a result of endogenous endonuclease activity. By contrast, the chromatin of thymocytes that retain normal density (whether treated or control) is of high molecular weight. Apoptotic cells, unlike those of normal density, show little or no incorporation of nucleosides and amino acids into macromolecules. These investigators were unable to detect populations of cells intermediate between apoptotic and normal in buoyant density, morphologic characteristics, chromatin cleavage, or leucine incorporation, but evidence is presented suggesting that thymidine and uridine incorporation may fall prior to development of apoptosis. PMID- 6289673 TI - Hormonal and hemodynamic responses to vena caval obstruction in fetal sheep. AB - To test the hypothesis that ACTH and vasopressin responses are quantifiable as functions of induced changes in central venous or arterial pressures, we produced various degrees of vena caval obstruction in fetal sheep (118--134 days gestation). In seven experiments, vena caval obstruction increased heart rate 18 +/- 7 beats/min and carotid arterial oxygen saturation 8.4 +/- 2.1%, but did not alter any measured vascular pressure or hormones. More severe vena caval obstruction (n = 10) decreased mean arterial pressure 13 +/- 2 mmHg, central venous pressure 1.3 +/- 0.3 mmHg, and heart rate 47 +/- 12 beats/min, and increased fetal plasma ACTH 1,047 +/- 448 pg/ml, cortisol 4.4 +/- 2.2 ng/ml, and vasopressin 47.9 +/- 24.2 pg/ml, but did not alter 11-deoxycortisol. The stimulus increased plasma cortisol (radioimmunoassay after chromatography) 100% and "corticosteroids" (radiotransinassay without chromatography) 20%, demonstrating the nonlinear relationship between these two variables. End-inflation plasma ACTH and vasopressin concentrations were significantly related to the induced decreases in mean arterial and central venous pressures, suggesting that the hormonal responses to vena caval obstruction were mediated by cardiovascular mechanoreceptors. Plasma vasopressin concentrations were linearly related to plasma ACTH concentrations (4 = 0.94; P less than 0.001), suggesting parallel release of the two hormones. PMID- 6289674 TI - Brown adipose tissue of rats with obesity-inducing ventromedial hypothalamic lesions. AB - Male and female Holtzman rats were made hyperphagic and obese with bilateral radiofrequency heat lesions of the ventromedial hypothalamic (VMH) area. When VMH rats were maintained at 28 degrees C, their brown adipose tissue (BAT) DNA, protein, and cytochrome oxidase contents were normal although more stored lipid was present, as judged from a threefold increase in wet weight. Thermogenic activity of BAT mitochondria was normal in male VMH rats, as judged from the unchanged level of guanosine diphosphate (GDP) binding (known to be a sensitive index of the functional activity of the thermogenic proton conductance pathway), and reduced in female VMH rats. When rats with VMH lesions were exposed to cold (4 degrees C for 24 h), the visible hyperemia of their BAT and normal large increase in mitochondrial GDP binding indicated normal thermogenic responsiveness. We conclude that the medial nuclei of the hypothalamus and associated afferent or efferent nerve tracts do not represent an essential central nervous system link for cold-induced, sympathetic-mediated activation of BAT thermogenesis. It is possible, however, that diet-induced, sympathetic mediated activation of BAT function and growth might require an intact VMH region because no enhancement of BAT mitochondrial function normally associated with hyperphagia was detected in these hyperphagic VMH-lesioned animals. PMID- 6289675 TI - Lack of solvent drag of NaCl and NaHCO3 in rabbit proximal tubules. AB - Using in vitro microperfusion of rabbit nephron segments we measured the effects of osmotically induced water flow on net transport of HCO3 and Cl. Measurements were made in superficial and juxtamedullary proximal convolutions and in superficial pars recta. In addition, measurements were taken in the presence and absence (hypothermia) of active transport. Using osmotic gradients of 25 mM raffinose in superficial and 50 mM in juxtamedullary segments, we observed increases in water flow equal to or greater than the normal rates of volume reabsorption observed in these tubule segments. However, there were no significant changes in HCO3 and Cl flux. This lack of significant solvent drag was seen both when osmotic water flow was in the lumen-to-bath direction and when osmotic flow was in the bath-to-lumen direction. The results of these studies suggest that solvent drag does not contribute significantly to NaCl and NaHCO3 reabsorption in proximal tubules. The lack of significant solvent drag of these salts can be interpreted as indicating either that osmotically induced transepithelial water flow in proximal tubules almost exclusively traverses transcellular pathways or that proximal tubule tight junction reflection coefficients for these salts are close to unity. PMID- 6289676 TI - Distribution of enzymes of adenylate and guanylate nucleotide metabolism in rat nephron. AB - In a previous study of discrete segments of rat nephron, we reported the levels of high-energy adenylate and guanylate phosphates to be highest in the distal straight and convoluted tubules. Those findings stimulated the study of the distribution of seven enzymes involved in the following metabolic pathways of these nucleotides [Formula: see text]. The patterns of distribution of enzymes in each pathway differed greatly. The phosphodiesterases, 1 and 2, were high in glomeruli and distal tubular segments and low in proximal segments. Adenylate kinase, 3, in contrast, was high in glomeruli, proximal segments, thick ascending limb of Henle, and distal convoluted tubules. Guanylate kinase levels, 4, however, were similar in all segments. The pattern of nucleosidediphosphate kinase, 5, was high in proximal convoluted, thick ascending limb, and distal convoluted tubules. The pattern of the degradative enzyme, 5'-nucleotidase, 6, whose levels were highest in proximal segments, was opposite from that of AMP deaminase, 7, highest in the distal nephrons. These dissimilar patterns underscore the extent of nephron heterogeneity. PMID- 6289677 TI - Redox state of cytochrome aa3 in isolated perfused rat kidney. AB - Optical spectroscopy was used to evaluate mitochondrial anoxia in the isolated perfused rat kidney by determining the percentage of cytochrome aa3 in the reduced state. Despite high levels of plasma flow (greater than 20 ml . g-1 . min 1) and venous PO2 (greater than 300 mmHg), 25--40% of cytochrome aa3 appeared to be in its reduced form. Agents that alter metabolism or transport primarily in renal cortex had little or no effect on the redox state of cytochrome aa3. These included 3-mercaptopicolinate, an inhibitor of gluconeogenesis; 2 tetradecylglycidic acid, an inhibitor of fat metabolism via carnitine-fatty acyltransferase, and acetazolamide. In contrast, a decrease in medullary transport induced by the loop diuretics bumetanide or furosemide produced an increase in cytochrome aa3 oxidation consistent with a decrease in transport related oxygen consumption in the hypoxic area. The results suggest that substantial portions of the kidney, particularly in the renal medulla, may normally operate on the brink of anoxia despite high PO2 in artery and vein. PMID- 6289678 TI - Does Na+-K+-atpase have any role in bile secretion? AB - In this review, I have attempted to summarize the evidence supporting the view that Na+-K+-ATPase (the enzymatic basis for the sodium pump) plays a key role in bile secretion. First, experiments in the isolated perfused liver, in isolated and cultured hepatocytes, and more recently in hepatocyte membrane vesicles strongly suggest that bile acid uptake by the hepatocyte is a secondary active transport that is energized by the Na+ gradient maintained by the Na+-K+-ATPase. Thus, Na+-K+-ATPase appears important in coupling the energy from ATP to transport activity, resulting in the so-called bile acid-dependent bile flow. Second, experiments with liver plasma membranes have shown a correlation between Na+-K+-ATPase activity in these preparations and the so-called bile acid independent bile flow. In view of the recognized sinusoidal-lateral localization of the Na+-K+-ATPase in the liver cells, several possibilities are proposed to explain how the enzyme activity could be coupled to ion transport and secretion into bile. These possibilities include Na+ movement from the intercellular space into the canalicular lumen through the paracellular pathway and movement of another ion using the Na+ gradient. PMID- 6289679 TI - Effect of phenolphthalein on monkey intestinal water and electrolyte transport. AB - To assess Na-K-ATPase inhibiton and prostaglandin synthesis stimulation as the mechanism of the secretory (cathartic) action of phenolphthalein in the primate, we investigated water and electrolyte transport and Na-K-ATPase levels in monkey intestine. Both jejunum and colon were studied with in vivo perfusion and in vitro Ussing chamber techniques. Water, Na, and Cl absorption was inhibited or secretion was induced by phenolphthalein (10(-3) M) in the jejunum and colon when the drug was present in the mucosal bathing (perfusion) solution. Serosal addition of phenolphthalein (10(-4) or 10(-3) M) induced Na and anion absorption in the jejunum but not in the colon. Phenolphthalein inhibited Na-K-ATPase activity in the test tube, but assays of intestine previously perfused or bathed in the drug showed no inhibiton. Indomethacin, in doses sufficient to inhibit prostaglandin synthesis in the intestine, inhibited the secretion induced by phenolphthalein in the jejunum but not in the colon. These inconsistencies cast doubt on the role of Na-K-ATPase inhibition or the role of prostaglandin synthesis stimulation in the mechanism of action of phenolphthalein. PMID- 6289680 TI - Quantitative electron microscope autoradiographs of 125I-cholecystokinin in pancreatic acini. AB - To morphologically evaluate the interaction of cholecystokinin (CCK) with its receptors on pancreatic acinar cells, we incubated isolated mouse acini at 37 degrees C with radioiodinated CCK and then prepared quantitative electron microscope autoradiographs. Specific binding of CCK to acini was one-half maximal at 2 min of incubation and maximal after 10 min. The cell-associated radioactivity was extracted and analyzed on Sephadex G-50. After 2 min, 90% of the total cellular radioactivity remained as intact CCK; after 30 min, the intact radioactivity decreased to 65% of total. At 2 min, the fraction of bound hormone that fixed to acini was 84% of total; this amount decreased to 78% after 30 min. Thus, the majority of radioactivity in the autoradiographs at both time points was intact CCK; however, at 30 min, a small amount was also degraded hormone. After both 2 and 30 min of incubation, silver grains were highly concentrated over the basolateral plasma membrane. A significant number of grains were in the cell interior at both time points, increasing from 13% of total grains at 2 min to 42% at 30 min. At both times, the largest fraction of internalized grains was localized over the endoplasmic reticulum. At 30 min, a significant concentration of CCK grains was observed over multivesicular bodies. The present study demonstrates, therefore, that CCK binds to specific receptors on the basolateral surface of pancreatic acinar cells. After binding, the hormone is internalized, locates predominantly on the endoplasmic reticulum, and is then degraded. PMID- 6289681 TI - Age-related effects of digoxin on myocardial contractility and Na-K pump in sheep. AB - The age-dependent effects of an acute nontoxic, positively inotropic dose of digoxin on myocardial monovalent cation active transport were determined in fetal, newborn, and adult sheep. Thirty-five lightly sedated, closed-chest animals were instrumented to record electrocardiogram, left ventricular (LV) pressure, and rate of change of LV pressure (LV dP/dt). Ouabain-inhibitable uptake of Rb+ (86Rb+) was measured in both right ventricular (RV) and LV slices from control animals and in animals infused with [3H]digoxin (0.04 mg/kg) sufficient to cause an increase in LV dP/dt without toxicity. Sixty minutes after digoxin, LV dP/dt increased 123% over base-line values in fetuses, 131% in newborns, and 165% in adult animals. RV and LV myocardial digoxin concentrations were similar in all groups. Rb+ active transport was significantly reduced in both RV and LV tissue from all animals 60 min after digoxin. Control animals showed no significant changes in contractility or Rb+ active transport among the control group of fetal, newborn, or adult sheep. Acute infusions of digoxin increased LV contractility in each age group and was accompanied by digoxin induced inhibition of myocardial Rb+ active transport. No age-related differences in the extent of Rb+ active transport among control or among digoxin-treated animals were observed under these experimental conditions. These studies suggest that the differential response to the therapeutic and toxic effects of digoxin in sheep of various ages does not reside in an age-dependent response of the myocardial sodium pump to digoxin. PMID- 6289682 TI - High plasma norepinephrine levels in patients with major affective disorder. AB - The authors found that patients with major affective disorder had higher levels of plasma norepinephrine and higher pulse rates (tachycardia) than healthy control subjects, but their blood pressures were normal. These measurements were similar in all three subgroups of patients with affective disorder--manic, bipolar depressed, and unipolar. Because norepinephrine is the primary neurotransmitter of the sympathetic nervous system, these data suggest sympathetic hyperactivity in the major affective disorders. This conclusion is compatible with recent speculation based on the effect of antidepressants on noradrenergic receptors and a failure of alpha-receptors to downregulate normally in patients with major affective disorder. PMID- 6289683 TI - Tubular carcinoma of the breast. Clinical and pathological observations concerning 135 cases. AB - Clinical and pathological features of 135 tubular carcinomas are discussed. Tumor size varied from 0.2 to 2.5 cm with a mean diameter of 0.9 cm. In situ carcinoma was found associated with tubular cancer in 86 cases (63.6%). In 82 of 86 (95.3%) it was of micropapillary/cribriform intraductal type. Twelve of 109 patients in whom axillary dissection was performed were found to have axillary metastases. Six patients (4%) developed recurrent or disseminated metastatic carcinoma during a mean follow-up period of 7.2 years. Two of these patients are dead of disease. Tubular carcinoma should be distinguished from microglandular adenosis, an uncommon form of sclerosing adenosis. PMID- 6289685 TI - Krukenberg tumor. PMID- 6289684 TI - New oncologic associations for the Epstein-Barr virus. PMID- 6289686 TI - Winter survival of blood-fed and nonblood-fed Culex pipiens L. AB - Comparisons were made between groups of Culex pipiens L. with different physiologic histories to test their ability to sucessfully overwinter under field conditions. On 14 December 1978, each group of mosquitoes was marked with a distinctive fluorescent dust and released inside an abandoned ammunition bunker at Fort Washington, Maryland. To insure that dead mosquitoes could be dissected and information obtained on their ovarian development, a sample of females from each group was also released into a plexiglass cage that was attached to the inside wall of the room. The physiologic histories of each group of mosquitoes were as follows: (a) "wild caught", those which had entered the bunker prior to the release date, (b) "lab-reared diapausing nonblood-fed," (c) "lab-reared diapausing blood-fed nongravid, " (d)"lab-reared diapausing blood-fed gravid," (e) "lab-reared nondiapausing nonblood-fed," and (f) "lab-reared nondiapausing blood-fed." By 8 March 1979, all of the lab-reared nondiapausing groups, of mosquitoes released in the room had died, whereas 15.7, 22.4 and 24.7% were recovered from the "lab-reared diapausing nonblood-fed," "lab-reared diapausing blood-fed" (gravid and nongravid) and "wild caught" mosquitoes, respectively. For the mosquitoes in the cage, only 0, 2.1 and 7.0% of the "lab-reared nondiapausing blood-fed," "lab-reared nondiapausing nonblood-fed" and "lab-reared diapausing blood-fed gravid," respectively, survived. This compared to 45.4, 56.8 and 58.0%, respectively, for the "lab-reared diapausing nonblood-fed," "lab-reared diapausing blood-fed nongravid" and the "wild caught" groups. These data provide evidence to support the theory that a significant number of diapausing Cx. pipiens which have taken a prehibernation (possibly viremic) blood meal do not develop eggs and can survive the winter at rates comparable to diapausing nonblood-fed mosquitoes. PMID- 6289687 TI - Chemotherapy of locally aggressive head and neck tumors in the pediatric age group. Desmoid fibromatosis and nasopharyngeal angiofibroma. AB - Our experience with the use of systemic chemotherapy in the management of locally aggressive head and neck tumors in the pediatric age group (desmoid fibromatosis and nasopharyngeal angiofibroma) is presented. Objective decreases in the size of tumors was found in all patients with desmoid fibromatosis was treated with chemotherapy before definitive surgical resection was performed. The changes noticed on examination correlated with the histologic and radiologic findings. Two patients with recurrent juvenile nasopharyngeal angiofibromas showed striking therapeutic improvement in their residual tumor mass as witnessed by radiographic studies and biopsies. Toxicity and side effects of the treatment are discussed. PMID- 6289688 TI - [Clinico-morphological characteristics of hydatidiform mole]. PMID- 6289689 TI - [Hormonal aspects of ovarian tumors]. PMID- 6289690 TI - [Treatment of hormonally active ovarian tumors]. PMID- 6289691 TI - [Current patholophysiologic and morphologic concepts of polycystic disease of the ovary]. PMID- 6289692 TI - A coupled optical enzyme assay phosphopentomutase. PMID- 6289693 TI - Determination of proton dissociation constants by ion-exchange high-performance liquid chromatography. PMID- 6289694 TI - Purification of human kidney angiotensin I converting enzyme using reverse immunoadsorption chromatography. PMID- 6289695 TI - The examination and quantitation of tissue cytosolic receptors for 2,3,7,8 tetrachlorodibenzo-p-dioxin using hydroxylapatite. PMID- 6289697 TI - Assay of calmodulin with Bordetella pertussis adenylate cyclase. PMID- 6289696 TI - The importance of silica type for reverse-phase protein separations. PMID- 6289698 TI - An improved method for the determination of oligonucleotide chain length using phosphodiesterase hydrolysis and ion-pair high-performance liquid chromatography. PMID- 6289699 TI - Sperm acrosin: liberation from the acrosome and activity of the free proteinase in the presence of nonoxinol-9. AB - The effects of the non-ionic detergent nonoxinol-9 and a vaginal contraceptive containing nonoxinol-9 on the liberation of the cellular proteinase acrosin from human spermatozoa and on the activity of the solubilized proteinase were investigated. Detachment of acrosin from the acrosome was observed in the presence of 0.01 to 1% nonoxinol-9 as assessed by activity assays and the gelatin substrate film technique. Activity of the free proteinase was modulated by nonoxinol-9 in a biphasic manner, i.e. lower concentrations of the detergent (0.001 to 0.1%) stimulated the activity of acrosin up to 180%, while higher concentrations of the detergent (1%) resulted in an extensive inhibition of acrosin (30% residual activity). It is demonstrated that local intravaginal nonoxinol-9 concentrations of 1% equalling the post-coital concentration of the intravaginal suppository Patentex oval when properly applied will result in an effective inhibition of the liberated sperm proteinase acrosin thus preventing unspecific proteolysis of the vaginal epithelia. PMID- 6289700 TI - Gallamine administered by combined bolus and infusion. AB - A technique combining an intravenous bolus and intravenous infusion regimen of gallamine based on its pharmacokinetics was developed to produce continuous relaxation during surgery. The combination of a bolus dose of gallamine, 2.5 mg/kg, and infusion, 0.8 mg/kg/hr, was tested in 11 patients. In 10 patients, surgery continued long enough to allow demonstration of an apparent plateau in the serum gallamine concentrations. At the cessation of the infusion, the mean gallamine concentration of 11.8 microgram/ml was associated with an average paralysis intensity of 92%. Pharmacokinetic analysis of the gallamine serum concentration-time data was fitted to a three-compartment model. In this study of 50- to 76-year-old patients, the most striking difference from other studies was that the elimination halflife averaged 247 minutes in this study whereas 128 to 141 minutes has been reported previously. PMID- 6289701 TI - Nitroprusside increases cyclic guanylate monophosphate concentrations during relaxation of rabbit aortic strips and both effects are antagonized by cyanide. AB - The authors have confirmed previous observations that sodium cyanide (CN-) partially reverses the vasodilator effects of sodium nitroprusside (SNP) on vascular smooth muscle. As tested on rabbit aortic strips contracted by norepinephrine (NE), the final tension is independent of the order of addition of reagents. In the same concentration, CN- alone had no effect on tension also as reported by others. The ED50 values for relaxation of aortic strips for a series of directly acting agonists ("nitric oxide vasodilators") were: sodium azide (N 3) 2.1 X 10(-7) M; SNP 2.7 X 10(-7) M; hydroxylamine (H2NOH) hydrochloride 2.5 X 10(-6) M; human nitric oxide hemoglobin (HbNO) 3.5 X 10(-6) M; and sodium nitrite (NO-2) 1.2 X 10(-4) M. In addition to SNP, CN- antagonized the vasodilator effects of N-3 and H2NOH, but it failed to reverse relaxation by HbNO, NO gas, NO 2 (as observed by us), glyceryl trinitrate, adenosine, or papaverine (as observed by others). The only change noted in cyclic-adenosine monophosphate (c-AMP) concentrations in aortic strips exposed to 1) NE, 2) NE + NO-2 or SNP, or 3) NE + NO-2 or SNP + CN- was an increase due to NE. The only statistically significant change noted in cyclic-guanosine monophosphate (c-GMP) concentrations exposed to 1) NE, 2) NE + NO-2 or 3) NE + NO-2 + CN- was also an increase due to NE. In contrast, SNP resulted in further increases in c-GMP after NE, and when cyanide was added, a significant decrease in c-GMP followed. These results are only partially consistent with a role for c-GMP in relaxation of vascular smooth muscle, but cyanide may become a useful tool for the study of mechanisms of action of the nitric oxide vasodilators. PMID- 6289702 TI - Herpes simplex virus from the lower respiratory tract in adult respiratory distress syndrome. AB - Herpes simplex virus (HSV) was found in the tracheobronchial secretions of 14 of 46 (30%) consecutive patients with the adult respiratory distress syndrome (ARDS). The HSV has not hitherto been associated with ARDS, and most previous reports of HSV in the lower respiratory tract have come from autopsy material. In the present study, the diagnosis during life was initially made by identification of the characteristic inclusion bodies of HSV in bronchial epithelial cells obtained from tracheobronchial aspiration. The presence of HSV in the lower respiratory tract was associated with the need for more prolonged respiratory support and an increased late mortality. PMID- 6289703 TI - Five-year longitudinal study of workers employed in a new toluene diisocyanate manufacturing plant. AB - The respiratory health of 277 workers in a new toluene diisocyanate (TDI) manufacturing plant was studied prospectively during 5 yr of exposure. Personal TDI monitors were used to continuously measure peak and 8-h time-weighted average (TWA) concentrations in over 2,000 samples. Longitudinal change in pulmonary function was assessed in 223 men in whom 3 or more data points allowed construction of individual slopes of annual change. Regression of annual change on smoking (pack-years), atopic status, and cumulative TDI exposure dichotomized at 68.2 parts per billion (ppb) months into low and high exposure groups showed significant effects of smoking on spirometric tests and lung volumes. After adjusting for pack-years of smoking, the 74 men in the high cumulative TDI exposure category had significantly larger declines in FEV1, %FEV, and FEF25--75% than did the 149 men in the low category. Annual change in FEV1 was then examined in 6 smoking-exposure categories: in never smokers, average annual decline was 38 ml/yr greater in those with higher cumulative TDI exposure. Current and previous cigarette smokers did not show this effect of cumulative TDI exposure. Analysis of FEV1 change by time above 20 ppb TDI yielded a similar result in never smokers, a 24 ml/yr excess average decline attributable to longer time above 20 ppb. In current cigarette smokers, those with longer time above 20 ppb had excess decline of 18 ml/yr (42 versus 24 ml/yr). our low and high cumulative exposure groups spent 2 and 15%, respectively, of their working time above 5 ppb TDI. The different health effects observed in these groups supports the NIOSH-recommended standard of 5 ppb TDI as an 8-h TWA. PMID- 6289704 TI - Identification of two monohydroxyeicosatetraenoic acids synthesized by human pulmonary macrophages. PMID- 6289705 TI - Elevation of the bronchoalveolar concentration of angiotensin I converting enzyme in sarcoidosis. AB - The concentration of angiotensin converting enzyme (ACE) and that of albumin (AIb) were assayed in the serum (SACE, SAlb) and in bronchoalveolar lavage fluid (LACE, LAlb). Three groups of patients were studied: 14 healthy volunteers (Group I), 45 patients with active sarcoidosis (Group II), and 7 patients with sarcoidosis in remission (Group III). The SACE in Group II (4,466 +/- 2,202 U/100 ml, mean +/- SD) was higher (p less than 0.001) than in Group I (2,470 +/- 547 U/100 ml) or in Group III (2,640 +/- 610 U/100 ml); LACE was higher in Group II (65.2 +/- 48.4 U/100 ml, p less than 0.001) than in Group I (21.1 +/- 14.7 U/100 ml), or in Group III (25.7 +/- 14.6 U/100 ml). The SAlb was found to be, respectively, 3,908 +/- 385 mg/100 ml, 3,982 +/- 965 mg/100 ml, and 3.613 +/- 222 mg/100 ml in Groups I, II, and III. The LAlb in Group II (8.2 +/- 6.2 mg/100 ml) was higher (p less than 0.01) than in Group I (2.5 +/- 1.4 mg/100 ml) or in Group III )4.1 +/- 1.0 mg/100 ml). The LACE in Group II increased with the number of alveolar lymphocytes, in nonsmokers (4 = + 0.56, df = 34, p less than 0.001) and in smokers (4 = + 0.88, df = 7, p less than 0.01). In the smokers in this group, LACE was higher with respect to the number of lymphocytes than in the nonsmokers. We conclude from this study (1) that the permeability of the alveolocapillary membrane to albumin and to ACE is increased in active pulmonary sarcoidosis, (2) that LACE increases during sarcoidosis and returns to normal when the disease is cured, and (3) that the concentration of ACE in alveolar fluid increases with tobacco use. PMID- 6289706 TI - Ceruloplasmin: plasma inhibitor of the oxidative inactivation of alpha 1-protease inhibitor. AB - When leukocyte lysosomal extracts are used as a source of elastase and are combined with a fraction of plasma containing sufficient alpha 1-protease inhibitor (alpha 1-Pi) to inhibit all but 30 to 40% of the elastase amidase activity, elastolysis occurs at 69% of the rate of the uninhibited elastase controls (0.125 M NaCl; pH, 6.5). Proteolysis of elastin requires the presence of NaCl. At pH 8.6, elastolysis is decreased to 30 to 40% of free elastase controls by 1.0 M NaCl. At pH 6.5, on the other hand, elastolysis is increased to 83% of the control values by these higher NaCl concentrations. The activity of human leukocyte myeloperoxidase is optimal at pH 6 to 6.5 and at NaCl concentrations between 0.25 and 1.0 M. Purified myeloperoxidase, alpha 1-Pi, and elastase, in the presence of NaCl and hydrogen peroxide, can reproduce this phenomenon at pH 6.5, suggesting that the occurrence of elastolysis in lysosomal extract-plasma mixtures may in part be a result of the oxidative inactivation of alpha 1-Pi by myeloperoxidase present in the lysosomal extract. Human ceruloplasmin, the major antioxidant of plasma, inhibits this myeloperoxidase-dependent reaction, without interfering either with free elastase activity or with the appearance of activity in plasma-lysosomal extract mixtures at pH 8.6. The "antioxidant" activity of ceruloplasmin is inhibited by azide. These results suggest that antioxidants such as ceruloplasmin may be an important determinant of lung defense in persons chronically exposed to oxidants. PMID- 6289707 TI - The effect of caffeine on diaphragmatic muscle force in normal hamsters. AB - The effect of caffeine on the tension generated by fresh and fatigued diaphragmatic muscle was examined in 32 Syrian hamsters. Studies were performed in vitro on innervated diaphragmatic muscle strips activated by electrical stimuli applied either directly or via the phrenic nerve. The degree of activation of the muscle was varied by altering the frequency of stimulation. When the stimulus frequency was low (less than 50 Hz), caffeine (1 mM and 5 mM) augmented the tension generated by fresh muscle in response to both phrenic nerve and direct stimulation in a dose-dependent fashion. At high stimulus frequencies (greater than 50 Hz), however, the response to caffeine was different in directly activated and phrenic-nerve-activated muscles. Caffeine augmented tension production at all stimulus frequencies in directly activated muscles, but decreased the tension developed in response to phrenic stimulation. The increase in tension elicited by caffeine during direct stimulation was relatively smaller at high compared with at low stimulus frequencies. Caffeine augmented peak tension by increasing the rate of tension development and prolonging the period during which tension increased. Similar effects were observed in muscle strips fatigued by prolonged activity. We conclude that caffeine augments tension production in both fresh and fatigued diaphragmatic muscle by a direct effect on the processes that activate contraction and that this effect is greatest when the muscle is minimally activated (low stimulus frequencies). Caffeine, however, may adversely affect muscle tension by impairing neuromuscular transmission. PMID- 6289708 TI - Role of smooth muscle alpha 1-receptors in nonspecific bronchial responsiveness in asthma. AB - It has been postulated that the wide range of bronchial hyperresponsiveness to nonspecific stimuli, such as methacholine, found in asthmatics could be due to increased alpha-adrenergic receptor activity in the airways. We examined alpha 1- receptor responsiveness by comparing the responses obtained after inhaling the alpha 1-agonist phenylephrine (2 to 32 mg/ml) with the responses obtained after inhaling buffered saline in 10 asthmatics in whom the provocation concentration of methacholine causing a 20% fall in the forced expiratory volume in one second (FEV1) ranged from 0.13 to 9.19 mg/ml. Subjects were pretreated with inhaled atropine (3 mg nebulized during tidal breathing) and inhaled propranolol (3 mg nebulized during tidal breathing) to exclude cholinergic and beta receptor activity influencing bronchial smooth muscle responsiveness. After pretreatment drugs, the mean value for FEV1 before phenylephrine inhalations did not differ significantly from that before the control inhalations of saline. There was no significant change in FEV1 after phenylephrine compared with that after saline, suggesting that bronchial smooth muscle alpha 1-adrenergic receptor activity was not present. We conclude that increased bronchial smooth muscle alpha 1-receptor activity is not the primary abnormality producing the variability between asthmatics in nonspecific bronchial hyperresponsiveness. PMID- 6289709 TI - Early detection of oxygen-induced lung injury in conscious rabbits. Reduced in vivo activity of angiotensin converting enzyme and removal of 5 hydroxytryptamine. AB - Changes in lung endothelial metabolic function, determined in vitro, have been proposed as sensitive indexes of hyperoxic lung damage. However, it is unclear whether these changes are also seen in vivo. We studied the possibility, using conscious rabbits in which jugular and carotid catheters had previously been placed under halothane anesthesia. Approximately 24 h later, test animals were exposed to normobaric hyperoxia (96 +/- 2%), while a second group was maintained in room air. Multiple indicator dilution methods were used to study (1) metabolism of 3H-benzoyl-phe-ala-pro (BPAP), a synthetic substrate for angiotensin converting enzyme (ACE), and (2) removal of 14C-5-hydroxytryptamine (5-HT) during a single transpulmonary passage in conscious animals. Determinations were made serially during exposure (room air or hyperoxia) or until death occurred in the oxygen-treated animals. Lungs of air-exposed animals hydrolyzed 81 +/- 2% of injected BPAP (0.1 to 0.15 nmoles) during a single passage. Percent metabolism was unaltered during the next 72 h. However, in test animals, ACE activity, as reflected by BPAP metabolism, was significantly reduced after 16 h of exposure to oxygen (77 +/- 2%, p less than 0.01) and continued to decrease to a nadir of 66 +/- 3% at 40 h. Single-pass lung uptake of 14C-5-HT (77 +/- 2%) was unchanged throughout the 72-h period in air-exposed rabbits. In test animals, 14C-5-HT removal decreased to 65 +/- 4% (p less than 0.01) after 24 h of oxygen exposure; 5-HT removal remained depressed compared with the 0 h control determination for the oxygen group at all subsequent measurement intervals. Light and electron microscopy of lungs from oxygen-exposed rabbits demonstrating reduced 5-HT removal and ACE activity at 24 h revealed normal endothelial and type I cell morphologic features. We conclude that exposure to 100% oxygen produced significant reduction in pulmonary 5-HT removal and BPAP metabolism prior to the onset of morphologic damage. PMID- 6289710 TI - Lung phagocyte recruitment and metabolic alterations induced by cigarette smoke in humans and in hamsters. AB - Cigarette smokers had an increased number of alveolar macrophages (AM) that had temporally related increases in oxidative metabolism in vitro compared with that in nonsmokers. The AM from young asymptomatic human cigarette smokers had a selective increase in superoxide anion (O2) release compared with those from nonsmokers. The AM from older smokers had a more intense, generalized enhancement of oxidative metabolism. Smoking hamsters had similar patterns of lung phagocyte recruitment and increased macrophage oxidative metabolism. The accumulation of AM within the alveolar ducts in smoking hamsters was strikingly similar to that seen in human smokers. The temporal patterns of smoke-induced changes in oxidative metabolism by AM from hamsters and humans were the same. Filtration of particulate constituents from cigarette smoke completely abrogated the distal airway inflammation and the metabolic alterations observed in smoking hamsters. PMID- 6289711 TI - In vitro susceptibility of Mycobacterium avium complex and Mycobacterium tuberculosis strains to a spiro-piperidyl rifamycin. AB - The spiro-piperidyl rifamycins are newly synthesized rifamycin S compounds. One of these compounds, LM 427, was tested in vitro against strains of the Mycobacterium avium complex and strains of M. tuberculosis; LM 427 inhibited 81.3% of 155 strains of the M. avium complex tested at a concentration of 1.0 microgram/ml compared with 5.8% inhibited by the same concentration of rifampin. Twenty-nine strains were resistant to both LM 427 and rifampin at 1.0 microgram/ml. Further testing of these 29 strains showed LM 427 inhibitory for all but 5 strains at 2.0 micrograms/ml and inhibitory for all but 1 at 5.0 micrograms/ml. Rifampin, on the other hand, inhibited none at 2.0 micrograms/ml and 11 strains at 5.0 micrograms/ml. The in vitro activity of LM 427 was also compared with rifampin by testing both compounds against M. tuberculosis at 1.0 microgram/ml. This comparison showed that all strains susceptible to rifampin were also susceptible to LM 427. However, 16 strains were susceptible to LM 427 and resistant to rifampin. The inhibition of drug-resistant mycobacterial species that cause pulmonary disease makes this compound an important consideration for future clinical studies. PMID- 6289712 TI - The value of technetium99m pyrophosphate scanning in the diagnosis of myocardial contusion. AB - Twelve patients who sustained significant blunt chest trauma and had abnormal electrocardiographic changes unrelated to hypotension, hypoxia, or pre-existing myocardial disease were studied prospectively during a one-year period. The patients were divided into two groups on the basis of serum creatine phosphokinase (CPK)/MB isoenzyme concentrations: Group A (six patients) had normal CPK/MB isoenzymes (less than 8 IU), and Group B (six patients) had elevated CPK/MB isoenzymes (greater than 8 IU). All patients underwent cardiac scanning with technetium99m pyrophosphate. All studies were interpreted as normal. These data suggest that technetium99m pyrophosphate scanning is not a reliable adjunctive test to confirm myocardial contusion in patients with significant blunt trauma of the chest. However, the diagnosis of myocardial contusion in such patients can be established by the presence of abnormal electrocardiographic changes associated with an elevation of the serum CPK/MB isoenzyme concentration. PMID- 6289713 TI - [Pancreatic resection in the treatment of infantile hypoglycaemia]. AB - Authors report four patients with hyperinsulinemic unremitting hypoglycaemia due to pancreatic nesidioblastosis. Onset was neonatal in three of them and at the end of the first year in the remaining one. After variable periods of only partially successful medical therapy, the four patients were operated and subtotal pancreatectomy was carried out. This alone was sufficient in one case, and in another one diazoxide made control possible. The other two children had a total pancreatectomy, and one of them has needed insulin ever since. Hypoglycaemia is under control in all children and only one has mental impairment. Diagnostic work-up and surgical techniques are described, and the need for a prompt operation when medical control is incomplete is stressed. Although apparently a blind and major undertaking, surgery is still the only way of preserving neuronal function in some selected cases. PMID- 6289715 TI - Influenza vaccines 1982-1983. Recommendation of the Immunization Practices Advisory Committee. Centers for Disease Control, Department of Health and Human Services; Atlanta, Georgia. PMID- 6289714 TI - Mycobacterium avium-intracellulare: a cause of disseminated life-threatening infection in homosexuals and drug abusers. AB - Five men developed disseminated infection with Mycobacterium avium intracellulare. These patients all lived in the New York City area and presented with their illnesses between January 1981 and September 1981; four were homosexual and one was an intravenous drug abuser. Four patients died. All five patients had defects in the cell-mediated immune response. The infections were characterized histopathologically by poor or absent granulomatous tissue reaction. Clinical isolates of M. avium-intracellulare from all five patients agglutinated commonly used antimycobacterial drugs. The spectrum of opportunistic infections among populations of homosexuals and drug abusers should be expanded to include disseminated disease due to M. avium-intracellulare. PMID- 6289716 TI - Toxic shock syndrome: a reprise. PMID- 6289717 TI - Precipitation of verapamil. PMID- 6289718 TI - Fast neutron therapy for orbital adenoid cystic carcinoma. PMID- 6289719 TI - Collagenolytic activity in keratoconus. AB - Corneal buttons were obtained from eight young adult patients with advanced keratoconus and were incubated in an organ culture system. The medium was assayed for collagenolytic activity using 14C-labeled reconstituted native collagen fibers as substrate. The assay was performed with and without prior collagenase activation with trypsin. The results indicated that the medium obtained from keratoconic corneas showed higher levels of collagenolytic activity compared with normal control corneas. We suggest that the level of collagenase activation may be a pathogenic factor in the development of keratoconus. PMID- 6289720 TI - Experience treating syndactyly. PMID- 6289721 TI - Combined embolization and surgery for craniofacial angiomas--report of a complex case. PMID- 6289722 TI - Evaluation of a simple colorimetric assay for serum angiotensin-converting enzyme: comparison with a new ion-pair liquid chromatography-assisted assay. AB - The development and validation of two different assays for serum angiotensin converting enzyme are reported. The first step in both analytical systems is based on enzymatic cleavage of the synthetic substrate, hippuryl-histidyl leucine, under conditions advocated by Cushman and Cheung. The hippuric acid released is then assayed either by colorimetry following an azlactone condensation reaction with an aromatic aldehyde, or by reversed-phase ion-pair high-performance liquid chromatography (HPLC). Both procedures reveal good linearity in the range 0-80 nmol/ml per min, with an inter-assay coefficient of variation of 6.2% for the colorimetric assay and 4.5% for the HPLC-assisted assay. Recovery values measured for the colorimetric method ranged from 97% to 102% and for the HPLC-assisted method from 98% to 101%. The colorimetric method is suitable for performance in small hospital laboratories since it can be carried out with simple analytical instrumentation. It is obvious nevertheless, that the HPLC-assisted assay reveals better validation criteria. The method is also simple and rapid and it has successfully been used in our laboratory for the last two years. PMID- 6289724 TI - [Mechanism of polyene antibiotic inactivation. Possible ways for levorin stabilization]. AB - Various means for levorin isolation were studied with the EPR method and approaches to stabilization of the antibiotic on storage under natural conditions were discussed. It was shown that formation of the radicals begins already at the first stage of the antibiotic isolation, i.e. during extraction from the mycelium. Treatment of the solvents with an inert gas or addition of antioxidants decreased the number of free radicals in a freshly isolated product. The antibiotic inactivation rate depended on the initial concentration of the free radicals and conditions of natural storage. The levorin stability increased when oxygen was thoroughly removed from the solvents at all isolation stages and the antibiotic was subsequently stored under conditions preventing any access of the air. The stabilizing effect was also observed when the oxidative effect of the amino sugar moiety on destruction of the polyenic chromophore during the antibiotic complex formation with respect to the amino group was decreased. PMID- 6289723 TI - The effect of 13-cis-retinoic acid on dibutylnitrosamine induced polyploidy changes in mouse urothelium. AB - The epithelium in the normal urinary bladder contains cells with diploid to octoploid DNA-content. The carcinogen dibutylnitrosamine (DBN), given subcutaneously in repeated doses causes a loss of polyploidy prior to cancer development. In this study the changes in polyploidy caused by DBN was followed by use of flow cytometry. 13-cis-retinoic acid did not prevent this loss of polyploidy, and did not affect the polyploidy in the normal urothelium. PMID- 6289725 TI - [Method of determining the antibiotic sensitivity of anaerobic microorganisms using standard disks]. AB - Three variants of the procedure for determination of antibiotic sensitivity in anaerobic microorganisms with the use of standard paper discs were developed. According to the first variant the solid nutrient medium is melted at 46 degrees C and mixed with the culture of the microbe being tested. The mixture is added to the cover of a Petri dish. When the medium becomes solid, antibiotic sensitivity discs are placed onto the agar surface. After that one more layer of the medium is added. The medium is allowed to solidify and some more medium is poured near the cover edge. Immediately after that the Petri dish is placed with its flat surface onto the agar layer in its cover. According to the first and second variants the mixture of the medium and culture is added to a Petri dish and immediately a transparent gas-proof polymer film of the dish size is placed onto the agar surface. Previously antibiotic paper discs or solutions are fixed on the films. THe incubation temperature for all three variants is 37 degrees C. The procedure allows one to observe the culture growth and to obtain the results earlier than in case the culture is incubated in an aerostate. The procedure is simple and saves labor and time. PMID- 6289726 TI - In vitro activity of Ro 13-9904, a new beta-lactamase-stable cephalosporin. AB - The minimal inhibitory concentration (MIC) of Ro 13-9904 against 245 clinical isolates was determined by an agar dilution method. The activity of Ro 13-9904 against most Enterobacteriaceae was similar to that of cefotaxime; it was slightly more active than cefotaxime against Proteus mirabilis, Providencia species, and Serratia marcescens, but slightly less active against Klebsiella species. Ro 13-9904 was twofold more active than cefotaxime and threefold more active than ticarcillin against ticarcillin-susceptible Pseudomonas aeruginosa, with a mean MIC of 7.2 micrograms/ml; isolates highly resistant to ticarcillin were inhibited by a mean MIC of 17.2 micrograms/ml. Ro 13-9904 was fourfold more active than ampicillin against susceptible Haemophilus influenzae and was equally active against beta-lactamase-producing isolates. Ro 13-9904 was highly active against pneumococci and moderately active (MIC, 4 micrograms/ml) against Staphylococcus aureus isolates, whether they were susceptible or resistant to penicillin G. Oxacillin-resistant S. aureus and Streptococcus faecalis were completely resistant to Ro 13-9904 (MIC, greater than 128 micrograms/ml). PMID- 6289727 TI - Susceptibility of shigellae to mecillinam, nalidixic acid, trimethoprim, and five other antimicrobial agents. AB - A total of 199 strains of shigella (1 Shigella dysenteriae, 15 S. boydii, 47 S. flexneri, and 136 S. sonnei) isolated in Malmo, Sweden, within a 3-year period (1977 through January 1980) were tested with the agar plate dilution method for susceptibility to commonly used and newer antimicrobial agents. Mecillinam, nalidixic acid, and trimethoprim had the best in vitro activity. S. flexneri dominated among strains resistant to three or more antimicrobial agents and were less susceptible to ampicillin, chloramphenicol, and doxycycline than other types studied. Sixty-four percent of the strains were resistant to sulfamethoxazole. In vitro, a synergistic effect with trimethoprim was shown only in strains susceptible to sulfamethoxazole. The amidinopenicillin mecillinam was highly active against shigellae. When resistance occurred, it was linked to ampicillin in 17 of 18 strains. The quinolines, here represented by nalidixic acid, might be the drugs of choice. PMID- 6289728 TI - Macrolide-lincosamide-streptogramin resistance patterns in Clostridium perfringens from animals. AB - Different patterns of resistance against commonly used macrolide, lincosamide, and streptogramin antibiotics were found in Clostridium perfringens of animal origin. The patterns were designated as (i) macrolide-lincosamide-streptogramin group B generalized resistance, (ii) macrolide-lincosamide generalized resistance, (iii) macrolide-lincosamide inducible resistance, and (iv) macrolide lincosamide-streptogramin low-level generalized resistance. The strains of the fourth pattern were able to inactivate pristinamycin and virginiamycin. The macrolide-susceptible strains showed a bimodal distribution of lincomycin and clindamycin susceptibility levels. The susceptible strains were inhibited by 0.25 micrograms of lincomycin per ml and 0.03 micrograms of clindamycin per ml. The low-level resistant strains were inhibited at concentrations of 2 to 4 micrograms of lincomycin per ml and 0.5 to 2 micrograms of clindamycin per ml. PMID- 6289729 TI - Stimulation of human and canine neutrophil metabolism by amphotericin B. AB - The effect of an antifungal agent, amphotericin B, on human and canine neutrophil metabolism was studied. Commercial preparations of amphotericin B in concentrations ranging from 5 to 100 micrograms/ml stimulated neutrophil chemiluminescence in the presence of 10(-8) M luminol. This response was blocked by 2-deoxyglucose, a metabolic inhibitor, and by the absence of extracellular calcium ions. Neither pure amphotericin B nor the solubilizing agent present in the commercial preparation, alone or in combination, stimulated neutrophil chemiluminescence. Commercial amphotericin B caused an increase in oxygen uptake by neutrophils but no detectable superoxide anion production. Neutrophils were injured by commercial amphotericin B, as shown by an increase in trypan blue dye uptake but not cell lysis. Binding of amphotericin B to neutrophil membrane sterol with a subsequent alteration in membrane configuration is the most likely cause of metabolic stimulation. PMID- 6289730 TI - Treatment of experimental Salmonella typhimurium infection with mecillinam and ampicillin. AB - The activities of mecillinam and ampicillin, alone and in combination, were evaluated in mice infected with the LT-2 strain of Salmonella typhimurium. The minimal inhibitory concentrations of mecillinam and ampicillin for this strain were, respectively, 6.2 and 0.4 microgram/ml of culture medium. In vitro synergy was demonstrated. CF-1 mice inoculated intraperitoneally with 10(4) colony forming units of the LT-2 strain were used in the therapeutic assessments. Treatment of subgroups with graded doses of the respective penicillins or their combination was initiated 24 h after inoculation and repeated at 6-h intervals for 5 consecutive days. Animals were observed during 21 days for mortality or sacrificed for quantitative cultures of spleen homogenates at the end of the treatment. Ampicillin in doses of greater than or equal to 0.03 mg and mecillinam in doses of greater than or equal to mg reduced mortality rates from 77% in the saline-treated controls to a range of 0 to 4% (P less than 0.05). The same doses of antibiotics also extended the median times to death and lowered significantly the means of splenic bacterial counts. When both drugs were combined in doses that were partially effective or subinhibitory alone, no synergistic effects were observed. These results showed that mecillinam and ampicillin given alone were effective in treating S. typhimurium infection but that combinations of the two drugs were not synergistic in controlling the course of infections. PMID- 6289731 TI - Comparative synergistic activity of cefoperazone, cefotaxime, moxalactam, and carbenicillin, combined with tobramycin, against Pseudomonas aeruginosa. AB - A broth dilution checkerboard synergy assay was used to assess the activity of cefoperazone, cefotaxime, moxalactam, and carbenicillin, in combination with tobramycin, against 38 strains of Pseudomonas aeruginosa. Synergy occurred significantly more often (P less than 0.001) when tobramycin was combined with cefotaxime (63%) than when it was combined with carbenicillin (26%), cefoperazone (21%), or moxalactam (18%). Of the 25 synergistically inhibited strains, the combination cefotaxime-tobramycin was synergistic against 24 and was the only synergistic combination against 10. Of six strains initially resistant to cefotaxime, five were susceptible to this agent (minimum inhibitory concentration, less than or equal to 32 micrograms/ml) when it was combined with tobramycin. Clinical trials are needed to determine the therapeutic efficacy of cefotaxime-aminoglycoside combinations in the treatment of serious Pseudomonas infections. PMID- 6289732 TI - Inconsistency of synergy between the beta-lactamase inhibitor CP-45,899 and beta lactam antibiotics against multiply drug-resistant Enterobacteriaceae and pseudomonas species. AB - Synergy between CP-45,899 and ampicillin or newer beta-lactam antibiotics against multiply drug-resistant Enterobacteriaceae and Pseudomonas species was inconsistent. In contrast, synergy between CP-45,899 and ampicillin against beta lactamase-producing strains of Haemophilus influenzae type b and Bacteroides fragilis was consistent and marked. PMID- 6289733 TI - Effect of ribavirin on Rous sarcoma virus transformation. AB - Ribavirin inhibited the expression of cellular transformation in normal rat kidney cells transformed by a temperature-sensitive mutant of rous sarcoma virus and chicken embryo fibroblasts infected with either a temperature-sensitive mutant or wild-type Rous sarcoma virus. Ribavirin also inhibited replication of the Rous sarcoma viruses in chicken embryo fibroblasts. The effect of ribavirin on cellular transformation was not permanent, as removal of the drug resulted in reversion to the transformed phenotype. The concentration of ribavirin necessary to inhibit the expression of cellular transformation was cytostatic for the cell lines used in this study. PMID- 6289734 TI - Ceftriaxone: in vitro studies and clinical evaluation. AB - The in vitro activity of ceftriaxone against 437 clinical isolates of gram negative bacilli was determined. Ceftriaxone was found to have high in vitro activity against Enterobacteriaceae, with the exception of Enterobacter cloacae. Ceftriaxone was only minimally active against Pseudomonas aeruginosa and Acinetobacter calcoaceticus. We evaluated the clinical efficacy and toxicity of ceftriaxone in 55 adult patients. Bacterial infection was confirmed by the isolation of etiological bacteria in 30 patients. Infectious disorders treated included 10 pneumonias, 13 urinary tract infections, and 7 soft tissue or bone infections. Pathogens identified were 25 isolates of gram-negative bacilli, 5 isolates of Staphylococcus aureus, 5 isolates of pneumococci, and 4 isolates of other streptococci. The overall efficacy of ceftriaxone was excellent. The clinical cure rate was 93%, and the bacteriological cure rate was 93%. A total of 30 adverse reactions were noted in 22 of 55 patients receiving ceftriaxone, but only one necessitated discontinuation of treatment. Adverse effects frequently noted were elevated hepatic enzymes (16%), thrombocytosis (16%), and eosinophilia (8%). Ceftriaxone is an effective and well-tolerated antimicrobial agent that appears promising for the treatment of serious gram-negative bacillary infections. PMID- 6289735 TI - Efficacy of a twelve-hourly ceftriaxone regimen in the treatment of serious bacterial infections. AB - Eighteen patients with 21 serious infections were treated with ceftriaxone, 1 g intravenously every 12 h, for a mean duration of 8 days. Eighteen gram-negative and two gram-positive organisms were isolated. Sites of infection included blood (three patients), urinary tract (six patients), respiratory tract (seven patients), biliary tract (three patients), ascitic fluid (one patient), and skin (one patient). Serum, bile, and ascitic fluid concentrations of ceftriaxone were in excess of the minimal bactericidal concentration required for the infecting organism in all cases. A bacteriological response was demonstrated in 94% of the infections. A clinical response occurred in four infections from which no pathogens were recovered. In one patient, ceftriaxone failed to eradicate a peritoneal infection due to Bacteroides fragilis. In two patients, superinfection with enterococci developed both during and after therapy. Systemic tolerance to ceftriaxone was excellent. PMID- 6289736 TI - Aminoglycoside-resistant gram-negative bacilli in a community hospital: comparative in vitro activity of cefotaxime, moxalactam, cefoperazone, and piperacillin. AB - All aminoglycoside-resistant gram-negative bacilli isolated during 1 year at a community hospital were tested for in vitro sensitivity to cefotaxime, moxalactam, cefoperazone, and piperacillin. The majority of Enterobacteriaceae were susceptible to all four antibiotics. Cefoperazone and piperacillin were the most active for pseudomonas aeruginosa, and activity for other nonfermentative gram-negative bacilli was variable. PMID- 6289737 TI - Comparative in vitro activities of azlocillin-cefotaxime and azlocillin tobramycin combinations against blood and multi-drug resistant bacterial isolates. AB - The in vitro activities of azlocillin and cefotaxime in combination and of azlocillin and tobramycin in combination against 100 blood isolates and 50 multidrug-resistant isolates were compared. With both combinations, antibacterial spectrums were complementary. Although synergy against individual strains was infrequently observed (except for azlocillin-cefotaxime against Streptococcus faecalis and azlocillin-tobramycin against Pseudomonas aeruginosa), 97% of blood isolates and 76% of multidrug-resistant isolates were susceptible to azlocillin cefotaxime, and 94% of blood isolates and 36% of multidrug-resistant isolates were susceptible to azlocyclin-tobramycin. PMID- 6289738 TI - Inhibition of herpes simplex virus replication by methyl daunosamine. AB - Methyl daunosamine inhibited the replication of herpes simplex virus type 1 in a dose-dependent manner. The growth of the host Vero cells was not affected by daunosamine levels that had significant antiviral activity (2.5 mM) but was inhibited by concentrations of 5 mM or greater. Methyl daunosamine appears to be unique among the sugars with antiviral activity because at antiviral concentrations it did not inhibit the glycosylation of macromolecules. PMID- 6289739 TI - Ceftriaxone (Ro 13-9904) therapy of serious infection. AB - Ceftriaxone (Ro 13-9904), a newly developed cephalosporin with a long half-life, was evaluated for efficacy and safety in 19 patients with serious infections. Underlying illnesses were present in 16 patients. Ceftriaxone was given intravenously every 12 h. Infections treated included gram-negative bacillary pneumonias (two cases), staphylococcal and streptococcal soft tissue-skeletal infections (six cases), spontaneous peritonitis (two cases), and complicated urinary tract infections (nine cases). Bacteremia was present in three patients. Microbiological and clinical cures were achieved in all but one case, although three patients with urinary infection had recurrences 6 weeks posttherapy. The only failure occurred in a patient with pneumonia who had a Pseudomonas aeruginosa isolated from sputum with an initial minimal inhibitory concentration of 4 micrograms/ml, but after 9 days of therapy, a repeat isolate had a minimal inhibitory concentration of 32 micrograms/ml. The minimal inhibitory concentrations for the other isolates ranged from less than or equal to 0.6 to 8.0 micrograms/ml. The mean peak plasma level of ceftriaxone was 99.9 micrograms/ml. The only side effects noted were drug fever in one patient, phlebitis in two patients, and thrombocytosis in four patients. PMID- 6289740 TI - Pharmacokinetics of ketoconazole in patients with neoplastic diseases. AB - Twenty-seven patients with advanced malignancies were given 200 mg of ketoconazole orally every 6 or 12 h. Blood samples were collected during these intervals and after the last dose to determine plasma concentrations and half lives. The mean plasma concentrations measured after the initial dose were 1.7 +/ 1.1 microgram/ml at 2 h, 0.9 +/- 0.2 microgram/ml at 6 h, and 0.7 +/- 0.4 microgram/ml at 8 h. Plasma concentrations rose significantly in patients on the every-6-h schedule. Concentrations were more variable in patients on the every-12 h schedule, and changes in mean plasma concentrations after 7 and 14 days were not significant. Half-lives ranged from 1.3 to 11.6 h in individual patients. The mean half-life for all patients studied was 3.7 +/- 0.6 h on day 1. The calculated area under the curve was 12.0 +/- 4.7 micrograms-h/ml on day 1; it increased after 7 and 14 days of administration (every-6-h schedule), suggesting plasma binding or wide drug distribution or both. Saturation of storage compartments is also suggested. Less than 1% of the administered dose was recoverable as active drug from the urine over 6 h. PMID- 6289741 TI - A new nucleoside analog, 9-[[2-hydroxy-1-(hydroxymethyl)ethoxyl]methyl]guanine, highly active in vitro against herpes simplex virus types 1 and 2. AB - A novel nucleoside analog, 9-[[2-hydroxy-1-(hydroxymethyl)ethoxy]methyl]-guanine (BIOLF-62), was found to have potent antiviral activity against herpes simplex virus types 1 and 2 at concentrations well below cytotoxic levels. For example, the Patton strain of herpes simplex virus type 1 was susceptible at concentrations 140- to 2,900-fold below that which inhibited cell division by 50%, depending upon the cell line used for assay. Different herpesvirus strains varied considerably in their susceptibility to the drug, as did results obtained with the same virus strain in different cell lines. BIOLF-62 compared favorably with 5-iodo-2'-deoxyuridine and acyclovir with respect to ratios of viral to cell inhibitory drug concentrations. Patterns of drug resistance to herpesvirus mutants suggested that the primary mode of action of BIOLF-62 is different from that of known antiviral compounds. Human adenovirus type 2, varicella-zoster virus, and Epstein-Barr virus were inhibited by this drug but at concentrations within the cell inhibitory range. Vaccinia virus and human cytomegalovirus were not inhibited at high drug concentrations. PMID- 6289742 TI - Herpes simplex virus variants restraint to high concentrations of acyclovir exist in clinical isolates. AB - Acyclovir (ACV) has been shown to inhibit the replication of herpes simplex virus (HSV) in vitro. We examined a wide variety of HSV clinical isolates for the presence of naturally occurring ACV-resistant (ACVr) variants. Although the ACV doses that inhibited 50% of these isolates were within the range of doses inhibiting 50% of the ACV-susceptible wild-type strains, we successfully isolated variants resistant to high ACV concentrations (25 to 75 microM) from each virion population even in the absence of prior drug exposure. Furthermore, we demonstrated, by fluctuation analysis of two encephalitis strains, that the ACVr variants were clonally distributed in the virus populations before exposure to ACV and did not result from rapid adaptation to ACV. All variants isolated after a single exposure to a high dose of ACV were true ACVr variants, as demonstrated by their plating efficiencies in the presence of ACV. We found that 36 and 50% of the ACVr variants of the two strains examined in detail displayed plating efficiencies in phosphonoacetic acid of greater than 0.1, possibly indicating that many of the ACVr variants contained alterations in the DNA polymerase gene locus. Because the distribution of ACVr variants in natural populations is relatively high (10(-4), these results suggest that selection of ACVr strains during ACV therapy is possible. PMID- 6289743 TI - Decreased susceptibility to 4'-deoxy-6'-N-methylamikacin (BB-K311) conferred by a mutant plasmid in Escherichia coli. AB - Escherichia coli MP6 contains a plasmid that encodes aminoglycoside 3' phosphotransferase II, which phosphorylates kanamycin and confers high-level kanamycin resistance, Amikacin is a minor substrate of this enzyme, but MP6 is susceptible to amikacin. Strain MP10 has a spontaneous mutation in the plasmid of MP6 that increases the aminoglycoside 3'-phosphotransferase II activity not only against kanamycin but also against amikacin. This mutation is also responsible for the appearance of resistance to amikacin in MP10. Resistance to 4'-deoxy-6'-N methylamikacin (BB-K311) by enzymatic modification has not been reported previously. As with amikacin, MP6 was susceptible to BB-K311 and its aminoglycoside 3'-phosphotransferase II did not phosphorylate this amikacin derivative appreciably. We found that the plasmid-borne mutation in MP10, however, localized by being cloned with a 3.7-megadalton HindIII fragment containing the aminoglycoside 3'-phosphotransferase II gene, resulted in increased phosphorylation of BB-K311 and resistance to it. Thus, the mutation distinguishing MP6 and MP10 has increased the activity of an existing aminoglycoside-modifying enzyme and produced new bacterial resistance to two previously minor substrates of the enzyme. PMID- 6289744 TI - Influence of high-fiber diet on bacterial populations in gastrointestinal tracts of obese- and lean-genotype pigs. AB - Bacterial populations from gastrointestinal tracts of genetically lean and obese pigs fed a low- or high-fiber diet (0 or 50% alfalfa meal, respectively) were enumerated with rumen fluid media and specific energy sources. Total culture counts in rectal samples declined 56 (P greater than 0.05) and 63% (P less than 0.05) in lean and obese animals, respectively, 3 weeks after feeding the high fiber diet. After 8 weeks, culture counts had risen and were similar to those obtained before alfalfa was fed (0 week). At slaughter, 12 to 17 weeks after feeding the high-fiber diet, total counts from rectal samples of lean pigs continued to rise and were 13% greater than the 0-week counts, whereas counts from obese animals declined 37% (P greater than 0.05). The number of cellulolytic bacteria in rectal samples of lean-genotype pigs fed the high-fiber diet increased 80 and 71% from 0 to 3 weeks and 3 to 8 weeks, respectively. This overall increases from 0 to 8 weeks in lean pigs was significant (P less than 0.05); however, these increases were not seen in obese pigs. These data suggest that the microflora is initially suppressed when exposed to a high-fiber diet and that later some adaptation takes place, apparently more so in lean than in obese pigs. When specific energy sources were used to delineate the distribution of different bacterial populations in the cecum, colon, and rectum, trends could be detected between high- and low-fiber diets. These data also support the concept that bacteria populations from different sites in the large bowel differ. PMID- 6289745 TI - Development of a shaker culture of Buffalo green monkey kidney cells: potential use for detection of enteroviruses. AB - Buffalo green monkey kidney cells were adapted to grow as shaker cultures. Replication of environmental and clinical isolates of poliovirus, coxsackievirus, and echovirus in these cultures was analyzed by plaque assay and compared with replication in Buffalo green monkey kidney cell monolayers and HEp-2 cell shaker cultures. Dose-response tests with various concentrations of Mahoney type 1 poliovirus indicated that Buffalo green monkey kidney cell shaker cultures could detect as little as 1 PFU in an inoculum of 0.2 ml. These data suggest that Buffalo green monkey kidney cell shaker cultures can be effectively used for the detection of small quantities of enteroviruses from environmental sources. PMID- 6289746 TI - Fate of virus in wastewater applied to slow-infiltration land treatment systems. AB - The removal of seeded coliphage f2 and indigenous enteroviruses from primary and secondary wastewaters applied by spray irrigation to sandy loam and silt loam soils in field test cells was examined. The amount of f2 recovered from 170-cm deep soil percolate samples taken over a 53-day period never exceeded 0.1% of applied virus levels and was usually below detection limits. Indigenous enterovirus levels in percolate waters also constituted only a small portion of those found in the wastewaters. At 10 days after seeding, f2 virus was present throughout the soil column but tended to accumulate around the soil core middepths. Coliphage f2 disappeared from the soil surface regions at a high rate, and by 53 days very little virus could be detected within the length of the soil columns. Sterilized soil core segments from different depths were studied to determine their virus adsorption capabilities when suspended in either wastewater, test cell percolate water, or distilled water containing divalent cations. The adsorptive capacity of Windsor and Charlton soils for poliovirus 1 and coliphage f2 increased greatly with the soil sample depth until leveling off at the midcore depths. Soil suspended in wastewater had the least virus adsorption capability for all depths studied. PMID- 6289747 TI - Histamine-producing bacteria in decomposing skipjack tuna (Katsuwonus pelamis). AB - Spoilage in skipjack tuna (Katsuwonus pelamis) was studied under controlled conditions by incubating whole, fresh fish in seawater at 38 degrees C, the optimum temperature for histamine formation. Bacterial isolates were obtained from the loin tissue of a decomposing tuna containing 134 mg of histamine per 100 g and a total anaerobic count of 3.5 x 10(5)/g after incubation for 24 h. Over 92% of the 134 isolates obtained were facultatively or obligately anaerobic bacteria. Eighteen isolates produced histamine in culture media containing histidine, and these were identified as Clostridium perfringens, Enterobacter aerogenes, Klebsiella pneumoniae, Proteus mirabilis, and Vibrio alginolyticus. Histidine decarboxylase activity of several isolates was measured in a tuna broth medium and with resting cells suspended in a buffered histidine solution. PMID- 6289748 TI - Location of lysyl residues at the allosteric site of fructose 1,6-bisphosphatase. PMID- 6289749 TI - Ca2+-ATPase from sarcoplasmic reticulum: acylphosphatase activity. PMID- 6289750 TI - The effect of ethanol and acetaldehyde on [14C]pantothenate incorporation into CoA in cultured rat liver parenchymal cells. PMID- 6289751 TI - Studies on the adenosine 3',5'-monophosphate-dependent protein kinase of Blastocladiella emersonii. PMID- 6289752 TI - Heparin inhibition and polyamine stimulation of a glycogen synthase kinase (PC0.7) from rabbit skeletal muscle. PMID- 6289753 TI - Purification of plant mitochondria by isopycnic centrifugation in density gradients of Percoll. PMID- 6289754 TI - Structure-function relationship in Escherichia coli initiation factors. Environment of the Cys residue and evidence for a hydrophobic region in initiation factor IF3 by fluorescence and ESR spectroscopy. PMID- 6289756 TI - Partial purification and characterization of cyclic AMP phosphodiesterases from Funaria hygrometrica. PMID- 6289755 TI - Beta-endorphin: thermodynamics of the binding reaction with rat brain membranes. PMID- 6289758 TI - Wilms's tumour and aniridia: clinical and cytogenetic features. AB - A survey carried out to detect children with aniridia/Wilms's tumour syndrome identified 8 living and 3 dead children. The incidence of aniridia was found to be 1 in 43 among Wilms's tumour patients in the UK. The clinical features included complete bilaterial aniridia, cataracts, glaucoma, mental retardation, hyperkinesis, hypospadias, and undescended testes. A high incidence of bilateral tumours (36%), male sex, presentation at a young age, and advanced maternal age appeared to be associated with the syndrome. The 8 living children each had a deletion on the short arm of chromosome 11. In contrast, although 2 patients with sporadic aniridia without Wilms's tumour had other malformations, neither had genitourinary anomalies, and the only additional problems in patients with familial aniridia were cataracts. Among 49 children with Wilms's tumour without aniridia ony one had bilateral tumours. No chromosome abnormalities were detected in patients with familial aniridia, nor were they detected in patients with Wilms's tumour without aniridia or in those with sporadic aniridia without Wilms's tumour. While many infants with the Wilms's tumour/aniridia syndrome are clinically diagnosable at birth, chromosome analysis using the elongated chromosome method is especially valuable to confirm the diagnosis in girls with sporadic aniridia and in boys who lack the genitourinary malformations. The presence of an 11p13 deletion confirms the diagnosis of the Wilms's tumour/aniridia syndrome and indicates a very high risk for the development of Wilms's tumour. PMID- 6289757 TI - Involvement of superoxide in the interaction of 2,3-dichloro-1,4-naphthoquinone with mitochondrial membranes. PMID- 6289759 TI - A contribution to the natural history of breast cancer. II. Precursors and lesions associated with small cancers of the breast. PMID- 6289760 TI - A contribution to the natural history of breast cancer. III. Changes in the basement membranes in breast cancers with stromal microinvasion. AB - Twenty-eight small breast cancers and 17 other breast cancer specimens from which appropriate semi-thin sections had been prepared were examined by electron microscopy and special staining techniques for alterations in the basement membranes and the basal lamina. In each case disruptions of the basement membranes were invariably observed at the point of initial invasion. Two types of invasion were noted: (1) bud-like protrusions of intraduct cancer with disruption of the basement membranes at the point of invasion; (2) extensive basement membrane defects with single cancer cell invasion. Our study confirms that invasive breast cancer originates in intraductal or lobular carcinoma in situ. PMID- 6289762 TI - Immunosuppression and experimental amoebiasis in guinea-pigs. PMID- 6289761 TI - Enzymatic release of microspheroids containing hydroxyapatite crystals from synovium and of calcium pyrophosphate dihydrate crystals from cartilage. AB - Incubation of minced, focally calcified, chondromatosis tissue obtained at operation from a patient with Milwaukee shoulder (rotator cuff defect and glenohumeral osteoarthritis associated with synovial fluid, hydroxyapatite crystals in microspheroidal masses, collagenase, and neutral protease) with partially purified mammalian synovial cell collagenase released masses of hydroxyapatite crystals of the same size as those originally found in the patient's synovial fluid. Incubation of mineral articular cartilage obtained from a shoulder joint at arthroplasty for a destructive arthropathy in a patient with generalised calcium pyrophosphate dihydrate (CPPD) crystal deposition with partially purified synovial cell collagenase freed CPPD crystals from their matrix. These data are compatible with a previously postulated mechanism linking microcrystals to destructive arthropathies, that is, crystal endocytosis by synovial cells stimulating collagenase secretion with subsequent enzymatic crystal "strip-mining', releasing additional crystals into the synovial fluid in a self-perpetuating cycle. PMID- 6289763 TI - Resistance to thyroid hormones. A disorder frequently confused with Graves' disease. AB - Five patients from two unrelated families were found to have goiter and elevated serum concentrations of thyroxine (T4) and triiodothyronine (T3) without symptoms or signs of hyperthyroidism. All had measurable concentrations of thyroid stimulating hormone (TSH), and in four who were tested, there was an increase in TSH concentration following thyrotropin releasing hormone (TRH) administration. We believe these five patients have general resistance to the effects of thyroid hormones and need elevated concentrations of T4 and T3 to maintain a eumetabolic state. Study of nuclear T3 receptors from cultured fibroblasts of one patient disclosed a normal equilibrium association constant and a maximal binding capacity that was greater than normal control values. These findings suggest that thyroid hormone resistance in this patient is not due to a decrease in either the affinity or the number of specific nuclear T3 receptors. This disorder can easily be confused with Graves' disease and result in inappropriate treatment for hyperthyroidism, as was the case in three of our patients. PMID- 6289765 TI - Hepatoma seen as an asymptomatic mediastinal mass. PMID- 6289764 TI - Apparent ketoconazole failure in candidal cholecystitis. AB - Fungal acalculous cholecystitis is a rarely recognized complication of disseminated candidal infections. We report such a case of systemic candidiasis that developed in a man who was diabetic while he was receiving broad-spectrum antibiotic therapy for an infected foot ulcer. Ketoconazole, a new imidazole antifungal agent, was used successfully to treat the systemic candidal infection but it failed to eradicate the fungus in the biliary tract. PMID- 6289766 TI - Comparison between viral RNA electrophoresis, ELISA and complement fixation techniques with electronic microscopy to demonstrate rotavirus. PMID- 6289767 TI - Biosynthesis of hexamannophosphoinositides in Mycobacterium smegmatis ATCC 607. AB - The biosynthesis of hexamannophosphoinositides in Mycobacterium smegmatis ATCC 607 was examined using labelled tri- and tetraacylated dimannophosphoinositides (PIM2(-3)F and PIM2(-4)F) as precursors, by in vivo and in vitro incorporation. Tetraacylated dimannoside was metabolically more active as compared to triacylated dimannoside and seems to be the precursor for the synthesis of hexamannophosphoinositides. PMID- 6289768 TI - [Infection eradication studies based on infectious bovine rhinotracheitis/infectious pustulous vulvovaginitis virus. 1. Modification of virus latency following natural IBR/IPV infection by subsequent immunization with Riems live IBR/IPV vaccine]. PMID- 6289769 TI - Search for retrovirus expression in men--failure to demonstrate retrovirus specific antigens in normal and malignant tissue. AB - Extracts for different normal and malignant human tissues were checked for retrovirus antigens by Sepharose bead immuno-fluorescence assay. No convincing evidence could be obtained for the presence of type D and mammalian type C virus antigens in the tissues tested by three different immunoassays. It is concluded from these results that such viruses have no ubiquitous distribution in human beings. PMID- 6289770 TI - Primary embryonal carcinoma and choriocarcinoma of the mediastinum. A case report. AB - Primary germinal tumors of the mediastinum are very rare. We studied a 67-year old man with a primary embryonal carcinoma and choriocarcinoma of the mediastinum. To our knowledge, this is the oldest patient described as having this lesion. The diagnosis of primary germinal tumor of the mediastinum should not be made unless the possibility of a primary testicular lesion has been eliminated by thorough clinical evaluation. The prognosis is very poor for patients with teratocarcinoma, embryonal carcinoma, or choriocarcinoma, despite aggressive therapy. PMID- 6289772 TI - Hypertrophic mononeuropathy. A report of two cases and review of the literature. AB - Two cases of hypertrophic mononeuropathy were studied. This entity is characterized grossly by focal enlargement of a single peripheral nerve, and microscopically by loss of nerve fibers, fibrosis, and formation of numerous onion bulbs. Our cases were diagnosed by exploratory surgery, biopsy, and light microscopy. Electron microscopic evaluation was performed in one case. Recognition of this entity by the surgical pathologist is important, so that it can be distinguished from generalized forms of onion bulb neuropathies, most of which give a much less favorable long-term prognosis. Hypertrophic mononeuropathy has usually resulted in irreversible local neurologic deficit but, unlike the polyneuropathies, in all the previously reported cases (as well as in our two cases to date) it has remained a localized process. PMID- 6289771 TI - Arthropathy in calcium pyrophosphate dihydrate crystal deposition disease. Pathologic study of 12 cases. AB - The pathologic features of calcium pyrophosphate crystal deposition disease (CPDD), particularly the synovial abnormalities, have not been adequately described or depicted in textbooks or journals; this report details the findings in 12 cases. Attention is drawn to the practical reasons for distinguishing CPDD arthropathy from other arthropathies, particularly osteoarthritis; clinical and gross pathologic features that should suggest CPDD arthropathy in cases that are not suspected preoperatively; and characteristics of the tophaceous deposits in CPDD. PMID- 6289773 TI - Epithelioid granulomas associated with hepatocellular carcinoma. PMID- 6289774 TI - Hypercalcemia and primary hepatic tumors. AB - We treated a case of hypercalcemia and primary liver tumor and reviewed a series of such cases treated at the Mayo Clinic (Rochester, Minn). Primary tumor of the liver was diagnosed in 192 patients (152 had hepatocellular carcinomas; 40, cholangiocarcinomas) between 1969 and 1980. Hypercalcemia of unknown cause was found in eight patients with hepatocellular carcinoma (5.3%) and seven with cholangiocarcinoma (17.5%). Five hypercalcemic patients had serum immunoreactive parathyroid hormone values consistent with ectopic hyperparathyroidism. An additional five patients had high serum calcium, low phosphate, and low chloride concentrations that met Lafferty's criteria for pseudohyperparathyroidism. Our results suggest that hypercalcemia associated with primary hepatic tumors is relatively common, and incidences vary according to the type of primary tumor. Hypercalcemia may be controlled when surgical excision of the primary tumor is possible. PMID- 6289775 TI - A comparison of cervical cytomegalovirus (CMV) excretion in gynaecological patients and post-partum women. AB - CMV was isolated from the cervix of 4.2 per cent of 191 gynaecological patients and from 9.8 per cent of 51 women post-partum; all patients were attending the same general practice clinic. The CMV excretion rate was particularly high in the early post-partum period decreasing to nearly normal levels as menstruation returned. Three of 14 (21.4 per cent) post-partum patients excreted CMV before menses had restarted whereas virus was isolated from only two of 36 (5.6 per cent) women who had returned to a normal menstrual cycle. Although this difference was not statistically significant, the excretion rate early post partum was significantly higher than in the gynaecological group (p less than 0.05). Five of seven excretors in the gynaecological group were in the first half of a menstrual cycle at the time of virus isolation thus suggesting that hormonal changes may lead to CMV reactivation in the genital tract. Other factors which may influence the presence of CMV in the genital tract of non-pregnant women are discussed. Three of four infant born to women excreting virus on the cervix post partum became infected with CMV. PMID- 6289776 TI - Characterization of a coronavirus isolated from rats with sialoadenitis. AB - A causative agent, provisionally designated as CARS, was isolated from the enlarged submaxillary gland of rat which was characterized as a sialoadenitis, using mouse-derived Balb/c3T3 clone A31 (3T3) cell culture. The virus could be propagated in 3T3 cell culture where it produced multinucleated giant cells and formed clear plaques. It was identified as a member of the coronavirus group from the following results: RNA content was suggested by the lack of the effect of cytosine arabinoside, the infectivity was sensitive to lipid solvents and inactivated at 56 degrees C for 5 minutes, the viral particle showed typical coronavirus morphology which was approximately 100 nm in diameter. Serologically, although CARS and sialodacryoadenitis virus (SDAV) actually belonged to the rat coronavirus group, some antigenic variations existed between these two agents in the results of both neutralization and complement-fixation tests using monovalent antisera. When inoculated intranasally into susceptible rats, CARS caused clinically and histologically overt sialoadenitis as observed in the natural outbreak, and retained its virulence for rats in several passages of mouse brain and even when cell culture was used, while rats which were inoculated with SDAV were asymptomatic. PMID- 6289777 TI - Lymphoid cell killing by human cytomegalovirus. AB - The interaction of human cytomegalovirus (CMV) with two replicating lymphoid cell lines, Ra-1 and MOLT-4F was studied. It was determined that while the cells exhibited viral receptors, the production of viral antigen and fully infectious virus could not be documented. Instead, cell death proportionate to the quantity of virus inoculated was seen. Neutralization of cell killing was effected with CMV-specific antiserum. In addition it was ascertained that cell death appeared to require a viral genomic function. This conclusion was based on the observation that UV irradiation of virus stocks reduced the cell killing capacity of the preparation. Infectivity of virus preparations was more sensitive to UV than the cell killing property. When compared on the basis of infectivity for fibroblasts and lymphoid cell killing capacity, pools of CMV with high infectivity had relatively less cell-killing capacity than low titered preparations. PMID- 6289778 TI - Immune response to a DNA free herpes simplex vaccine in man. AB - A DNA-free subunit herpes simplex virus (HSV) vaccine was administered to 15 volunteers without past evidence of HSV infection and to 25 patients with severe recurrent HSV infection. The immune response to the vaccine in these patients was compared to the immunological status of 20 non-vaccinated control patients with recurrent HSV infection. The vaccine elicited antibody and cell-mediated immunity (CMI) in the 15 subjects without past evidence of HSV infection and this response was similar to that observed after a natural infection. Among the 25 patients who were suffering from recurrent HSV infection the vaccine elicited complement dependent cytotoxic antibodies in 13 of these patients who did not possess these antibodies and increased significantly the titers of these antibodies in the 12 other patients. The vaccine gave a significant increase of the titers of the other specific antibodies as well as the level of cell-mediated immunity. The increase of the immunity level in these latter patient was not due to normal variations since in the non-vaccinated control group the antibody titers and CMI remained stable during the same period of time. PMID- 6289779 TI - Ultrastructural study on type C virus particles in a human cord T-cell line established by co-cultivation with adult T-cell leukemia cells. PMID- 6289780 TI - Morphological method for estimation of simian virus 40 infectious titer. AB - The cytomorphologic method previously reported for titration of adenoviruses has been employed for estimating the infectious titer of simian virus 40 (SV 40). Infected cells forming intranuclear inclusions were determined. The method examined possesses a number of advantages over virus titration by plaque assay and cytopathic effect. The virus titer estimated by the method of inclusion counting and expressed as IFU/ml (Inclusion Forming Units/ml) corresponds to that estimated by plaque count and expressed as PFU/ml. PMID- 6289781 TI - Recovery of latent herpes simplex virus from human trigeminal nerve roots. PMID- 6289782 TI - [Rheumatoid synovitis (problems of immunomorphology and immunopathogenesis)]. AB - The immunopathogenesis of rheumatoid synovitis (RS) is assumed to be as follows: antigen X interacts with the organism of subjects genetically predisposed to rheumatoid arthritis (RA) as a result of which the immune response regulation is disturbed ("immunological discomfort") leading to changes in proliferation and differentiation of immunocompetent cells immunologically manifested by disturbed ratios of lymphocyte subpopulations and morphologically by immune inflammation of synovial membranes, i.e. RS. Immune inflammation in RS is a systemic, chronic, persisting, recurrent, self-sustained process based on profound changes in qualitative and quantitative aspects of immunological homoeostasis. The results of the authors' own studies indicate that in the inflammation field (eluates of synovial membrane and synovial fluid) rearrangement of lymphocyte subpopulations is more marked than in the peripheral blood of patients with RA. The quantitative deficiency in eluates of synovial membrane and peripheral blood of T gamma lymphocytes having the suppressive-cytotoxic function confirms the important role of immunoregulation disturbance in the pathogenesis of RA. PMID- 6289783 TI - [Morphology of thrombolysis as affected by a plasmakinase preparation]. AB - The effect of a new thrombolytic preparation, plasmakinase, on experimental coagulates and thrombi was studied morphologically. Histological and electron microscopic studies revealed changes in the structure of fibrin fibers in the form of separation of fibers into individual protofibrils and subsequent degradation into globular particles. There were disorders in the fibrin association with blood cells, washing out of partially hemolyzed red blood cells into the bloodstream, decrease in the size of thrombi and restoration of the vessel lumen. PMID- 6289784 TI - [Ovarian thecoma with amyloidosis]. AB - A case of diagnosis of a variety of ovarian thecoma, a thecoma with amyloidosis of the stroma, in a patient with fibromyoma of the uterus is described. Brief clinical data, characteristics of the tumor morphology including polarization microscopy and histochemistry, and differential diagnosis of forms of amyloidosis are presented. The presence of amyloid in the thecoma is considered to reflect the capacity of the tumor theca-cells of mesenchymal origin to produce amyloid. PMID- 6289785 TI - [Incidental syringohydromyelography: report of a case]. AB - The radiological findings of syringomyelography are described in a five years old patient who underwent a percutaneous injection of opaque contrast medium (Lipiodol) into the intraspinal syringomyelic cavity. The clinical picture, the usual diagnostic methods and the place of syringomyelography for the diagnosis of syringomielia are discussed. PMID- 6289786 TI - An experimental study of lithium and dental caries in the rat. AB - Lithium chloride, given via the drinking water, was tested for its effect on dental caries. In a preliminary experiment lithium at concentrations of 0, 5, 20, 40, 60, 125 and 250 microgram/l showed a significant dose-related effect by ANOVA with the lowest smooth-surface scores at 20 microgram Li. Lithium was also used at 20 microgram/l with and without fluoride at 5, 10 and 20 mg/l. No greater effect on caries was seen of the lithium-fluoride combination than fluoride alone. Using 0, 5, 20 and 40 microgram Li/l, and larger groups of rats, a trend for lower caries was seen but the variance of the data was too large to be statistically significant. Despite a number of epidemiologic studies associating low caries prevalence with lithium, animal model research has not demonstrated a clear reduction in caries due to lithium added to the drinking water. PMID- 6289787 TI - Differential response of type I and type II cyclic AMP-dependent protein kinases in submandibular gland of isoproterenol-treated rats. AB - The cytosolic protein kinase isozymes were studied in relation to the secretion of saliva and the proliferative process induced by isoproterenol. At the same time as salivary secretion began, cytosolic protein kinase was activated and, in the absence of cyclic AMP, increased to a maximum level at 10 min after administration of the drug. The active state of the enzyme was maintained for 1-2 h. Chromatographic analysis of the cytosol enzyme revealed that in an activated state type II kinase easily dissociates into the catalytic and regulatory subunits while type I kinase hardly dissociates. Thus we conclude that type I kinase, at least in submandibular gland, is less dissociable in the presence of cyclic AMP than type II kinase but highly active in a ternary complex (R2-4 cAMP C2). The presence of cyclic AMP-independent protein kinase, which co chromatographed with protein kinase II, was demonstrated by use of a heat-stable inhibitor from bovine muscle, which totally inhibited the cyclic AMP-dependent enzymes, but stimulated the cyclic AMP-independent protein kinase. The level of protein kinase I decreased to its lowest value, about half that of the control value, at 10 h after isoproterenol injection and then increased slightly at 15 h, whereas type II kinase remained virtually unchanged, suggesting that the two protein kinase isozymes are independently regulated in the proliferating cells. PMID- 6289789 TI - Pattern of cortisol release in sheep following administration of synthetic ACTH or imposition of various stressor agents. AB - This paper describes experiments with sheep, in which changes in plasma cortisol, after imposition of various stressor agents, were compared to the changes following administration of synthetic ACTH. The influence of stress associated with shearing, yarding, oestrogen administration (30 microgram oestradiol benzoate, i.v.), feeding and fasting on the plasma concentration of cortisol was monitored in four mature Merino ewes. They were placed in the experimental environment for 21 days before monitoring began. The ewes were treated in pairs, each pair being visually and acoustically isolated from the other. One pair of ewes was exposed to the particular stress and the remaining pair acted as their own controls. The treatments were reversed 2 days later. In the second part of the experiment, 0, 0.01, 0.1, 1.0 or 10.0 i.u. synthetic ACTH were injected as an intravenous bolus, after endogenous secretion had been suppressed by administration of synthetic glucocorticoid. All blood samples were taken via an indwelling jugular catheter. A comparison of cortisol release--estimated from a plot of cortisol in plasma versus time--following imposition of various stressor agents and administration of synthetic ACTH, allowed stress to be defined in terms of synthetic ACTH equivalents. The most severe stress was associated with shearing (0.84 i.u. synthetic ACTH equivalents), less stress was imposed by yarding and handling (0.45 i.u.), and there appeared to be no effect attributable to feeding, fasting or oestrogen administration. The similarity in the pattern of cortisol release following ACTH administration offers the possibility of defining acute, but not chronic, stress in terms of ACTH equivalents. PMID- 6289788 TI - Sensory transduction and neuronal transmission as related to ultrastructure and encoding of information in different labyrinthine receptor systems of vertebrates. AB - Mechano-electric transduction and neuronal transmission were studied in sensory systems ascending from and descending to single receptor cells of the labyrinth organs in submammalian vertebrates. The animals were young crocodiles (Caiman crocodilus), geckos (Gekko gecko, Tarentola mauritanica), and turtles (Pseudemys scripta elegans, Chinemys reevesii). Intracellular receptor potentials from the apical region of the hair cell (or from the ciliary surface) were recorded in the ampullar, macular, and papillar sensory cells. These single-cell responses are, within limits, proportional to stimulus amplitude, frequency, or phase and are bidirectional in that they show depolarization by kinociliopetal stereociliar displacement and hyperpolarization by kinociliofugal displacement. Synaptic potentials (presynaptic from the basal region of the hair cell, postsynaptic from the contacting nerve endings) were recorded in the utricular, saccular, and lagenar neuroepithelia with electron-optic localization of the in situ fixed microelectrode tip. As local excitatory or inhibitory processes, respectively, they follow the stimulus and receptor potential with latency and with nonlinear distortion. Action potentials (spikes), as synchronized by the excitatory synaptic potentials, were recorded from single nerve fibers or bipolar cells, related to ampullar, macular, or papillar receptor units. Unit responses and synaptic potentials were recorded from the first, second, and following centripetal and central neurons of the ascending systems, or from neurons of the descending systems in the brain stem or from centrifugal neurons. Such records were achieved during adequate mechanical or acoustical stimulation of the different receptor systems, with additional electrical stimulation, uni- or bilaterally. Thus, the influence of centripetal-centrifugal bilateral interaction on the receptor functions was measured, as inhibition or disinhibition, respectively. The input-output relations of these sequential stages of information transfer were plotted as histograms of different types, as characteristic curves, power spectra, or by correlation operations, with or without feedback, from the different systemic levels.U PMID- 6289790 TI - The characterisation and pathogenicity of porcine enteroviruses isolated in Victoria. AB - Approximately 23 viruses were isolated from healthy pigs, pigs with encephalitis, and in cases of reproductive failure. Five viruses were identified as enteroviruses and a total of 10 isolates were shown to cross-react serologically to varying degrees. Twenty viruses were neutralised by a reference antiserum of serotype 8 porcine enterovirus. Intracerebral inoculaton of colostrum-deprived piglets with 2 of the characterised viruses caused lesions of encephalomyelitis which were not induced by oral infection. Intrafoetal inoculation of 2 sows with one characterised faecal isolate caused foetal death and abortion, but no adverse effects followed oral dosage. PMID- 6289791 TI - Functioning adrenocortical tumour in a dog. AB - Naturally-occurring hyperadrenocorticism was diagnosed in an 11-year-old female Dachshund with signs of polydipsia, polyuria, pendulous abdomen, weakness, depression and lethargy, and laboratory test abnormalities comprising lymphocytopaenia, eosinopaenia, hypercholesterolaemia and increased plasma alkaline phosphatase concentration. While awaiting hormonal test results, an adrenocorticolytic drug (o,p'-DDD) was administered for 14 days, during which the patient deteriorated. Hormonal assays suggested a functioning adrenocortical tumour, but the poor condition of the patient precluded adrenalectomy. An adrenocortical carcinoma with hepatic metastases was found at necropsy. PMID- 6289792 TI - Classification of the orbiviruses. Confusion in the use of terms bluetongue virus, bluetongue-like virus, bluetongue-related virus and the overall nomenclature. PMID- 6289793 TI - Aetiological aspects of chronic arthritis (big knee) in goats. PMID- 6289795 TI - Nictitating membrane reflex of the frog: effects of paraorbital shock and body temperature. PMID- 6289794 TI - Lithium chloride-induced avoidance of a conspecific odor: effect of prior exposure to the conditional stimulus. PMID- 6289796 TI - Preliminary observations on Hofbauer cells in short-term culture. PMID- 6289798 TI - Application of a live varicella vaccine to hospitalized children and its follow up study. PMID- 6289797 TI - Neuromuscular blocking in acutely tetanus intoxicated mice. AB - The effects of tetanus toxin on neuromuscular transmission of mice in acute intoxication produced by intravenous injection of a large amount of the toxin were examined by (1) recording the phrenic nerve impulses, the electromyograms (EMGs) of the diaphragm and the electrocardiograms, and (2) the evoked EMGs of the gastrocnemius muscle in response to electrical stimulation of the sciatic nerve. The evoked EMGs of the gastrocnemius muscle were analyzed in terms of kinetic and tonic components by their different latencies. Just before death of animals, the EMGs of the diaphragm appeared with some delay relative to the corresponding phrenic discharges. Finally, the EMG of the diaphragm disappeared even in the presence of phrenic discharge, but cardiac electrical activities continued. The amplitudes of the evoked EMGs of the gastrocnemius muscle invariably became low before death, but the muscle action potential could be recorded by direct muscle stimulation for several minutes after death. The latencies of the evoked EMGs were constant until about the middle of the survival time when the latencies suddenly became prolonged. The longer latency was the same as that of the tonic action potentials. Thus, in acutely tetanus-intoxicated mice, neuromuscular transmission was blocked rapidly and the kinetic component of the muscle was blocked earlier than the tonic component. PMID- 6289799 TI - Protective effect of immediate inoculation of a live varicella vaccine in household contacts in relation to the viral dose and interval between exposure and vaccination. AB - The protective effect on 45 household contacts of immediate inoculation of a live varicella vaccine was examined in relation to the viral dose and days after exposure to varicella. The viral doses used were 300 PFU to 15,000 PFU, and the intervals between exposure and vaccination were 0 to 5 days. All 30 contacts given viral doses of 800 to 15,000 PFU within 3 days after exposure were protected from clinical varicella. With lower viral doses of 300 to 600 PFU within 3 days after exposure, 6 to 10 contacts developed rashes 13 to 25 days after vaccination, but all showed mild symptoms. When vaccine was given 5 days after exposure, even with a dose of 1,500 to 4,000 PFU, typical varicella symptoms appeared 6 to 9 days after vaccination. These results suggest that vaccination of household contacts with viral doses of over 500-800 PFU within 3 days after exposure is effective in preventing manifestation of clinical varicella and that clinical symptoms may be modified to a mild form by vaccination with even a lower viral dose when given within 3 days after exposure. PMID- 6289800 TI - An Fc-receptor activity of plasma membranes from guinea-pig peritoneal polymorphonuclear leucocytes. AB - Plasma membranes prepared from guinea-pig peritoneal polymorphonuclear leucocytes showed an immune-complex-binding activity that corresponded well with the activity in intact cells. The characteristics of this activity were reversible binding, dependence on the Fc portion of antigen-complexed IgG (immunoglobulin G), competition with aggregated IgG and independence from energy metabolism. These results support the conclusion that the binding activity found in the isolated plasma membranes is an Fc-receptor activity of guinea-pig peritoneal polymorphonuclear leucocytes. The activity showed Kd = 6.7 x 10(-8) M-IgG and maximum binding of 17 pmol of IgG/mg of membrane protein when measured with an immune complex of alpha-amylase and homologous guinea-pig anti-(alpha-amylase) IgG. Inhibition of the Fc-receptor activity by a series of various salts indicated the contribution of the hydrophobic interaction to the binding. Inhibitory effects of salts or metal-chelating reagents on the Fc-receptor activity were also observed on superoxide generation by these cells induced by the immune complex, suggesting a role of the Fc receptor as the immune-complex binding site responsible for the initiation of superoxide generation. PMID- 6289801 TI - Alternative-substrate inhibition and the kinetic mechanism of the beta galactoside/proton symport of Escherichia coli. AB - The effects of competing alternative substrates on the rate of uptake by galactoside/proton symport were investigated. These experiments produced a decrease in apparent maximum velocity with increased alternative-substrate concentration that cannot be accounted for by a simple ordered mechanism. This, together with non-linearities in the variation of the apparent kinetic constants with alternative-substrate concentration, can be accounted for by a random mechanism for galactoside and proton binding. PMID- 6289802 TI - Effects of adenosine, 2-deoxyadenosine and N6-phenylisopropyladenosine on rat islet function and metabolism. AB - Adenosine (1.0-100 mum). N(6)-phenylisopropyladenosine (0.1-10 mum) and 2 deoxyadenosine (10 mm) all produced a dose-dependent inhibition of glucose stimulated insulin release. The inhibition of glucose-stimulated insulin release by adenosine and N(6)-phenylisopropyladenosine was abolished by 3-isobutyl-1 methylxanthine (0.1 mm), whereas 2-deoxyadenosine inhibited insulin release even in the presence of 3-isobutyl-1-methylxanthine. These adenosine nucleosides also inhibited the release of insulin induced by 4-methyl-2-oxopentanoate (20 mm), dl glyceraldehyde (30 mm) and l-leucine (20 mm). Adenosine (10 mum). N(6) phenylisopropyladenosine (10 mum) and 2-deoxyadenosine (10 mm) did not inhibit insulin biosynthesis or [U-(14)C]glucose oxidation at concentrations of the nucleosides that gave maximal inhibition of insulin release. However, adenosine, 2-deoxyadenosine and N(6)-phenylisopropyladenosine produced marked inhibition of the glucose-stimulated increases seen in islet cyclic AMP accumulation. Similar to its effects on insulin release, 3-isobutyl-1-methylxanthine (0.1 mm) antagonized the inhibitory effects of cyclic AMP accumulation produced by adenosine and N(6)-phenylisopropyladenosine, but had no effect on the inhibition of cyclic AMP accumulation seen with 2-deoxyadenosine. These results show that adenosine and its specifically modified analogues, 2-deoxyadenosine and N(6) phenylisopropyladenosine, are strong inhibitors of insulin release from rat islets, a function that appears to be the consequence of their ability to inhibit the accumulation of cyclic AMP. It is proposed that the B cells, in common with many other tissues, may possess two different sites at which adenosine nucleosides interact to produce their biological effects; these are the so-called ;P' and ;R' sites first described by Londos & Wolff [(1977) Proc. Natl. Acad. Sci. U.S.A.74, 5482-5486]. PMID- 6289804 TI - Labelling of prolyl hydroxylase tetrameric subunits in freshly isolated chick embryo tendon cells and in certain chick-embryo tissues in vivo. AB - The labelling of the subunits of prolyl 4-hydroxylase tetramers was studied in freshly isolated chick-embryo tendon cells and in chick-embryo tissues. In the former both the alpha- and beta-subunits of the tetramer were labelled during a 4 h labelling and 2 h chase period, although the radioactivity in the beta-subunit was much lower than in the alpha-subunit. The corresponding subunits of the enzyme from 12-day chick-embryo cartilaginous bone and heart were labelled in 7 h, again the beta-subunit much less than the alpha-subunit, the ratio of radioactivity in the beta-subunit to that in the alpha-subunit (beta/alpha radioactivity) being 0.20 and 0.32 respectively. The beta/alpha-radioactivity then increased almost linearily with time between 7 and 24 h, by 9.5-fold in the cartilaginous bone and 3-fold in the heart, and beta/alpha-radioactivity values above 1.0 were reached. The free beta-subunit-size protein (the beta'-protein), which is also present in cells, had been labelled quite heavily by 7 h. The beta/alpha-radioactivity at 7h, determined in four tissues with different ratios of prolyl hydroxylase tetramers to total immunoreactive protein (tetramer percentage), was low in tissues with a high tetramer percentage. It is thus proposed that only a minor fraction of the beta'-protein must be processed to the tetrameric beta-subunit and utilized in the synthesis of the prolyl 4-hydroxylase tetramers. PMID- 6289803 TI - 1 alpha,25-Dihydroxycholecalciferol and a human myeloid leukaemia cell line (HL 60). AB - Human promyelocytic leukaemia cells (HL-60) can be induced to differentiate into mature granulocytes in vitro by 1 alpha,25-dihydroxycholecalciferol [1 alpha,25(OH)2D3], the active form of cholecalciferol. The differentiation associated properties, such as phagocytosis and C3 rosette formation, were induced by as little as 0.12 nM-1 alpha,25(OH)2D3, and, at 12 nM, about half of the cells exhibited differentiation on day 3 of incubation. Concomitantly the viable cell number was decreased to less than half of the control. Among various derivatives of cholecalciferol examined, 1 alpha,25(OH)2D3 and 1 alpha,24R dihydroxycholecalciferol were the most potent in inducing differentiation, followed successively by 1 alpha,24S-dihydroxycholecalciferol, 1 alpha hydroxycholecalciferol, 25-hydroxycholecalciferol and 24R,25 dihydroxycholecalciferol. A cytosol protein specifically bound to 1 alpha,25 (OH)2D3 was found in HL-60 cells. Its physical properties closely resembled those found in such target tissues as intestine and parathyroid glands. 1 alpha,25(OH)2D3 bound to the cytosol receptor was transferred quantitatively to the chromatin fraction. The specificity of various derivatives of cholecalciferol in inducing differentiation was well correlated with that of their association with the cytosol receptor. These results are compatible with the hypothesis that the active form of cholecalciferol induces differentiation of human myeloid leukaemia cells by a mechanism similar to that proposed for the classical concept of steroid hormone action. PMID- 6289805 TI - Non-co-ordinate development of beta-adrenergic receptors and adenylate cyclase in chick heart. AB - We have studied the properties of beta-adrenergic receptors and of their interaction with adenylate cyclase in the chick myocardium during embryogenesis. Between 4.5 and 7.5 days in ovo the number of receptors determined by (-) [3H]dihydroalprenolol ([3H]DHA) binding is constant at approx. 0.36 pmol of receptor/mg of protein. By day 9 the density decreases significantly to 0.22 pmol of receptor/mg of protein. At day 12.5--13.5 the number was 0.14--0.18 pmol of receptor/mg of protein. This number did not change further up to day 16. The same results were obtained with guanosine 5'-[beta, gamma-imido]triphosphate (p[NH]ppG) added to the assay mixtures. There was no significant change in receptor affinity for the antagonist [3H]DHA between days 5.5 and 13. Despite the decrease in numbers of beta-adrenergic receptors, there was no change in basal, p[NH]ppG-, isoprenaline- or isoprenaline-plus-p[NH]ppG-stimulated adenylate cyclase activity between days 3 and 12 of development. We conclude that beta adrenergic receptors and adenylate cyclase are not co-ordinately regulated during early embryonic development of the chick heart. Some of the beta-adrenergic receptors present very early in the ontogeny of cardiac tissue appear not to be coupled to adenylate cyclase since their loss is not reflected in decreased activation of the enzyme. PMID- 6289806 TI - Effect of 5'-deoxy-5'-isobutylthioadenosine on putrescine uptake and polyamine biosynthesis by chick embryo fibroblasts. AB - The effect of 5'-deoxy-5'-S-isobutylthioadenosine (SIBA) on polyamine biosynthesis has been studied by using cultured chick embryo fibroblasts. It has been shown that the drug inhibits the uptake of [14C]putrescine and its conversion into labelled spermidine or spermine. The inhibitory effect is reversed by removing the inhibitor after exposing the cells to the drug for 24 h. SIBA also caused a significant decrease in cellular spermine levels and an accumulation of putrescine. These changes are reversed by removing the inhibitor. SIBA had the same effect on chick embryo fibroblasts transformed by Rous sarcoma virus; a decrease in cellular spermine levels in SIBA-treated cells was observed. In all the experiments SIBA caused a reduction in the spermine/putrescine and spermidine/putrescine ratios. It is suggested that SIBA is not only an inhibitor of transmethylation but also interferes with polyamine biosynthesis, probably by blocking aminopropyltransferase. PMID- 6289807 TI - Structural role of the tyrosine residues of cytochrome c. AB - The tertiary structures of horse, tuna, Neurospora crassa, horse [Hse65,Leu67]- and horse [Hse65,Leu74]-cytochromes c were studied with high-resolution 1H n.m.r. spectroscopy. The amino acid sequences of these proteins differ at position 46, which is occupied by phenylalanine in the horse proteins but by tyrosine in the remaining two, and at positions 67, 74 and 97, which are all occupied by tyrosine residues in horse and tuna cytochrome c but in the other proteins are substituted by phenylalanine or leucine, though there is only one such substitution per protein. The various aromatic-amino-acid substitutions do not seriously affect the protein structure. PMID- 6289808 TI - Evidence for catalytic dismutation of superoxide by cobalt(II) derivatives of bovine superoxide dismutase in aqueous solution as studied by pulse radiolysis. AB - By using the technique of pulse radiolysis to generate O2-., it is demonstrated that Co(II) derivatives of bovine superoxide dismutase in which the copper alone and both the copper and zinc of the enzyme have been substituted by Co(II), resulting in (Co,Zn)- and (Co,Co)-proteins, are capable of catalytically dismutating O2-. with 'turnover' rate constants of 4.8 X 10(6) dm3.s-1.mol-1 and 3.1 X 10(6) dm3.s-1.mol-1 respectively. The activities of the proteins are independent of the pH (7.4-9.4) and are about three orders of magnitude less than that of the native (Cu,Zn)-protein. The rate constants for the initial interaction of O2-. with the Co-proteins were determined to be (1.5-1.6) X 10(9) dm3.s-1.mol-1; however, in the presence of phosphate, partial inhibition is apparent [k approximately (1.9-2.3) X 10(8) dm3.s-1.mol-1]. To account for the experimental observations, two reaction schemes are presented, involving initially either complex-formation or redox reactions between O2-. and Co(II). This is the first demonstration that substitution of a metal into the vacant copper site of (Cu,Zn)-protein results in proteins that retain superoxide dismutase activity. PMID- 6289811 TI - Limitations of commonly used spectrophotometric assay methods for phosphoenolypyruvate carboxykinase activity in crude extracts of muscle. AB - Phosphoenolpyruvate carboxykinase activity in crude extracts of muscle has frequently been determined by using a continuous spectrophotometric method, which is shown to grossly overestimate enzyme activity. NADH oxidation attributed to phosphoenolpyruvate carboxykinase activity in the assay is due to lactate production. Under the normal assay conditions. Na+ ions stimulate pyruvate kinase, providing pyruvate for lactate formation by lactate dehydrogenase and sufficiently to account for most of the observed NADH oxidation. PMID- 6289810 TI - Deoxynucleotide-interconverting enzymes and the quantification of deoxynucleoside triphosphates in mammalian cells. AB - We have demonstrated that methanol extracts of human cells are heterogeneous with regard to content of dNDP (deoxynucleoside diphosphate) and dNMP (deoxynucleoside monophosphate) kinases. The presence of these enzymes can affect the reliability of techniques used to measure intracellular pools of deoxynucleotides. An optimized extraction procedure and enzymic assay for dNTP species in haematopoietic cells are described which provide sensitivity to measure 0.1 40pmol of dATP, dTTP and dGTP, and 1.0-40pmol of dCTP. The extraction and assay give linear results with (2.5-15)x10(6) nucleated cells and (0.1-1.5)x10(9) red blood cells. Under these conditions, extracts equivalent to ~0.5x10(6) nucleated haematopoietic cells catalyse the phosphorylation of 0-8% of dNDP and dNMP standards to dNTP and incorporate them into deoxynucleotide polymer under circumstances where 100% of an equimolar dNTP standard would be incorporated. By contrast, extracts of 0.4x10(6) HeLa cells totally converted dADP, dTDP and dGDP into dNTP with subsequent polymerization. Conversion of dCDP was somewhat less efficient. The results demonstrate conclusively that the activities of deoxynucleotide interconverting enzymes differ in different types of human cells. They can interfere with assay of nucleotides, but may not do so in many types of cell extracts. In particular, dNTP concentrations can be measured in human haematopoietic cells after extraction with 60% (v/v) methanol and are not artificially elevated by deoxynucleotide interconversions. It is apparent that extraction and assay procedures for measurement of dNTP species should be analysed for each cell type in order to minimize contaminating enzyme activities and ensure accuracy of dNTP quantification. PMID- 6289812 TI - Specific binding of 1,25-dihydroxycholecalciferol in human medullary thyroid carcinoma. AB - A specific 1,25-dihydroxycholecalciferol-binding protein has been detected in high-salt cytosols prepared from human medullary thyroid carcinomas. The binding protein had the same equilibrium dissociation constant (Kd = 0.17 +/- 0.05 nM; n = 4) and sedimentation coefficient on sucrose gradients (3.7S) as than seen in established vitamin D target tissues. This protein was not detected in normal thyroid cytosols, which may reflect the low proportion of C-cells within the gland. PMID- 6289809 TI - Fructose 2,6-bisphosphate 2 years after its discovery. PMID- 6289813 TI - A specific L-tri-iodothyronine-binding protein in the cytosol fraction of human breast adipose tissue. AB - 1. Binding of l-tri-[(125)I]iodothyronine to the cytosol fraction of normal human female breast adipose tissue was investigated by the charcoal adsorption method. Equilibrium of binding was reached after 120s at 25 degrees C. 2. The l-tri [(125)I]iodothyronine-binding component is a protein; this was confirmed by experiments in which binding was totally lost after heating the cytosol fraction for 10min at 100 degrees C and in which binding was diminished after treatment with proteolytic enzymes and with thiol-group-blocking reagents. The binding protein was stable at -38 degrees C for several months. 3. It displayed saturability, high affinity (apparent K(d) 3.28nm) and a single class of binding sites. 4. High specificity for l-tri-iodothyronine and l-3,5-di-iodo-3' isopropylthyronine was observed, whereas other iodothyronines were less effective in displacing l-tri-[(125)I]-iodothyronine from its binding site. 5. The binding of the hormone by the cytosol fraction did not show a pH optimum. 6. When cytosol fractions of adipose tissue from different females were subjected to radioimmunoassay for the determination of thyroxine-binding globulin a value of 0.304+/-0.11mug/mg of cytosol protein (mean+/-s.d., n=4) was obtained; the mean concentration in plasma was 0.309+/-0.07mug/mg of plasma protein (mean+/-s.d., n=3). 7. The K(a) value of 6.3x10(8)m(-1) of l-tri-[(125)I]iodothyronine for binding to plasma, the similar thermalinactivation profiles of binding and the reactivity to thiol-group-blocking reagents were some properties common between the binding components from the cytosol fraction and plasma. 8. These results suggest that the cytosol fraction of human female breast adipose tissue contains thyroxine-binding globulin; the protein that binds l-tri-[(125)I]iodothyronine with high affinity and specificity appears to be similar to thyroxine-binding globulin. PMID- 6289814 TI - ADP-ribosyl-protein conjugate subclasses in various tissues. Specific influence of thyroid hormone on liver conjugates. AB - The amounts of endogenous mono (ADP ribose)-protein conjugates and their hydroxyl amine-sensitive and hydroxylamine-resistant subfractions in various tissues of the mouse were determined and compared with poly (ADP-ribose) conjugates. Total mono-(ADP-ribose) conjugates did not correlate with poly (ADP-ribose) residues or the cellularity of a tissue, but were related to the protein content and the amounts of NAD+ +NADH. The hydroxylamine-resistant subfraction, which in liver is mainly associated with the mitochondria [Adamietz, Wielckens, Bredehorst, Lengyel & Hilz (1981) Biochem. Biophys. Res, Commun. 101, 96-103], did not correlate with the tissue content of cytochrome c oxidase. In hypothyroid mice hydroxylamine resistant mono (ADP-ribose) conjugates of the liver were increased by a factor of two while the hydroxylamine-sensitive conjugates did not change significantly under these conditions. Upon administration of thyroxine the hydroxylamine resistant subfraction returned to normal. PMID- 6289815 TI - Ornithine decarboxylase and polyamine levels are reduced in CHO cells deficient in cAMP-dependent protein kinase. PMID- 6289816 TI - Forskolin as a novel lipolytic agent. PMID- 6289817 TI - Localization of a site interacting with human platelet receptor on carboxy terminal segment of human fibrinogen gamma chain. PMID- 6289818 TI - Hypomethylation of HeLa cell DNA and the absence of 5-methylcytosine in SV40 and adenovirus (type 2) DNA: analysis by HPLC. PMID- 6289819 TI - Properties of Na+ channels in fibroblasts. PMID- 6289821 TI - Metabolism of carbon tetrachloride to electrophilic chlorine by liver microsomes: exclusion of cytochrome P-450 catalyzed chloroperoxidase reaction. PMID- 6289820 TI - The alpha and beta subunits of lamb kidney Na,K-ATPase are both glycoproteins. PMID- 6289822 TI - Thiol-dependent lipid peroxidation. PMID- 6289823 TI - Paradoxical activation by glucose of quinine-sensitive potassium channels in the pancreatic B-cell. PMID- 6289824 TI - Synthesis of collagen, collagenase and collagenase inhibitors by cloned human gingival fibroblasts and the effect of concanavalin A. PMID- 6289825 TI - Photolabeling of glucose-sensitive cytochalasin B binding proteins in erythrocyte, fibroblast and adipocyte membranes. PMID- 6289826 TI - Adenosine kinase from Trypanosoma cruzi. PMID- 6289828 TI - Dibutyryl cyclic AMP treatment mimics ovariectomy: new genomic regulation in mammary tumor regression. PMID- 6289827 TI - Inhibition of membrane-bound adrenodoxin-adrenodoxin reductase activity by puromycin and cycloheximide, which antagonize the adrenal steroidogenic action of ACTH. PMID- 6289829 TI - Phosphorylation with cyclic adenosine 3':5' monophosphate-dependent protein kinase renders bovine cardiac troponin sensitive to the degradation by calcium activated neutral protease. PMID- 6289830 TI - (Na+,K+)ATPase levels in preadipocyte cell lines established from genetically obese and non-obese mice. PMID- 6289831 TI - Ligation of restriction endonuclease-generated DNA fragments using immobilized T4 DNA ligase. PMID- 6289832 TI - Ion conducting pores induced by oligo-L-alanine. PMID- 6289833 TI - Heterogeneity of the calcium-dependent phosphatidylinositol-phosphodiesterase of rat liver kidney, as revealed by column chromatofocusing. PMID- 6289834 TI - Induction of ethylene biosynthesis in tobacco leaf discs by cell wall disesting enzymes. PMID- 6289835 TI - The dissociation properties of native C1. PMID- 6289836 TI - Kinetic properties of rat hepatic prolactin receptors. PMID- 6289837 TI - Vinculin phosphorylation by the src kinase: inhibition by chlorpromazine, imipramine and local anesthetics. PMID- 6289840 TI - Epidermal growth factor receptors in fetal and maternal rabbit lung. PMID- 6289838 TI - A multifunctional cyclic nucleotide- and Ca2+-independent protein kinase from rabbit skeletal muscle. PMID- 6289839 TI - Photoaffinity labeling of pituitary gonadotropin releasing hormone receptors during the rat estrous cycle. PMID- 6289841 TI - Preparation and characterization of a stable half met derivative of type 2 depleted Rhus laccase: exogenous ligand binding to the type 3 site. PMID- 6289842 TI - Anion binding to oxidized type 2 depleted and native laccase: a spectroscopically effective model for exogenous ligand binding to the type 3--type 2 active site. PMID- 6289843 TI - Prostaglandins and cannabis--IX. Stimulation of prostaglandin E2 synthesis in human lung fibroblasts by delta 1-tetrahydrocannabinol. AB - Preliminary data [S. Burstein and S. A. Hunter, Biochem. Pharmac. 27, 1275 (1978)] showed that cannabinoids at levels of 1 microM or greater elevated the concentrations of prostaglandins in cell culture models. Further study [S. Burstein and S. A. Hunter, J. clin. Pharmac. 21, 240S (1981)] led to the suggestion that this effect was due to a stimulation of phospholipase A2 resulting in the release of free arachidonic acid which was then partly converted into the prostaglandin(s) normally synthesized by the particular target system. The present report gives detailed data on the cannabinoid-induced synthesis of prostaglandin E2 by te WI-38 fibroblast derived from human lung. The effect could be blocked by pretreatment with mepacrine, a phospholipase inhibitor, and aspirin, a cyclooxygenase inhibitor. These findings lend support to the hypothesis that some of the in vivo actions of the cannabinoids are due to modulations in prostaglandin synthesis at various tissue sites. PMID- 6289844 TI - Effect of thyroid status on beta-adrenergic receptor, adenylate cyclase activity and guanine, nucleotide regulatory unit in rat cardiac and erythrocyte membranes. PMID- 6289845 TI - Urinary metabolites of delta 1-tetrahydrocannabinol in man. AB - The metabolism of delta 1-tetrahydrocannabinol (delta 1-THC) was studied in man after oral administration. Twelve acidic metabolites were isolated from human urine and identified by gas chromatography-mass spectrometry. One additional metabolite was tentatively identified. Hitherto, only one of the metabolites (delta 1-THC-7-oic acid, ) has been identified in man. Three of the metabolites have not been identified in any species before. delta 1-THC-7-oic acid (1) corresponding to 27% of the urinary metabolites was the most abundant metabolite. 4"-Hydroxy-delta 1-THC-7-oic acid (6) was the second most abundant monocarboxylic acid. The remaining eleven monocarboxylic acids predominantly oxidized at C-7 and at C-2" and C-3" of the side-chain, unsubstituted as well as monohydroxylated, were present in small amounts. PMID- 6289847 TI - [Amphicrine, mucoid and argyrophile tumors of the breast. Histochemical and ultrastructural study of 2 cases]. AB - The authors report two cases of mucoargyrophilic carcinomas of the breast, occurring in 73 and 81 year old women. Both tumors consist of a central mucoid carcinoma surrounded by solid areas with lobular or endocrinoid features. Most cells showed an abundant eosinophilic granular cytoplasm or a mucinous appearance. The histochemical reactions (Grimelius-Alcian Blue, Grimelius Mucicarmine) and the ultrastructural study of the peripheral areas disclosed several cell types: poorly differentiated or mucinous cells were present in both cases; but most of the cells were argyrophilic and contained in their hyaloplasm numerous dense core "neurosecretory" granules; such cells revealed many microfilaments arranged in bundles or in whorls. Amphicrine cells were easily demonstrated by histochemical staining and their cytoplasm showed both endocrine granules and mucinous vesicles. These features suggest a possible relationship to other argyrophilic tumors of the breast. The histogenesis of these tumors is still little known. PMID- 6289846 TI - Comparative pharmacokinetics of oestradiol, oestrone, oestrone sulfate and "conjugated oestrogens" after oral administration. AB - The time course of free oestradiol and oestrone as well as of total (free and conjugated) oestrone was determined in plasma of women after oral ingestion of 3.75 mg conjugated equine oestrogens, or 9.74 micromol of oestrone sulfate, oestrone, and oestradiol, respectively. All these oestrogen preparations led to transiently increased plasma oestrogen levels which fell to normal values after 48 h. The main difference observed between administration of free oestrone or oestradiol and of conjugated oestrogens (oestrone sulfate or equine oestrogens) was a much more protracted influx of oestrogens from the intestine into the plasma compartment, with a tendency to more sustained plasma levels, if conjugated oestrogens were administered. There was a consistent discontinuity in plasma oestrogen levels 10--12 h after oral ingestion of all the preparations examined indicating enterohepatic circulation. Comparison of "areas under the curve" obtained with the present preparations to similar previous studies on ethinyl oestradiol indicated that the bioavailability of the non-ethinyl oestrogens is by more than one order of magnitude less than that of ethinyl oestradiol after oral administration. The ratio of oestrone/oestradiol was the highest with free oestrone and similar among the other three preparations indicating an increased metabolism of sulfoconjugated oestrone to oestradiol after oral application when compared with free oestrone. PMID- 6289848 TI - Evidence that opioid receptors in the substantia nigra pars reticulata are relevant in regulating the function of striatal efferent pathways. AB - A tonic activity in the electromyogram (EMG) was induced in conscious rats by injections of morphine either systemically or into the caudate nucleus. This activity was antagonized by injections of naloxone into the substantia nigra pars reticulata. These findings suggest that opioid receptors in this brain area are relevant in regulating the function of striatal efferent pathways. PMID- 6289849 TI - [Epidemic of viral hepatitis in a mountain community of western Sicily]. AB - Epidemiological investigations were carried out during and after a viral hepatitis A outbreak in a mountain town, Geraci Siculo, in western Sicily. Blood samples were obtained through finger prick or by venipuncture from patients, their family contacts and from healthy school children of different age groups (3 to 15 year old). Serum markers for hepatitis A (anti HAV and IgM anti HAV) and hepatitis B (HBsAg and anti HBs) were tested. All but two of 36 cases, which occurred from August through March with a peak in November and December were under 15; 33 of them, in which laboratory data were available, were due to HAV. Seroepidemiological investigations, performed in December 1979 and in May 1980, have shown a low prevalence of anti HAV antibodies: 11.5% only of the children tested were positive in the first sampling and 22% in the second one. Also prevalence of HBV markers was low, as compared with similar observations carried out previously in other different area of Western Sicily. PMID- 6289850 TI - [Quantitative determination of diazepam in pharmaceutical preparations by PMR]. PMID- 6289851 TI - Effect of lead on physical state of erythrocytes and model membranes. AB - Lead has an high affinity for erythrocyte and influences the membrane bound enzymes activities but its interactions with biological membranes is still unclear. We have investigated the effect of lead as free ion on physical state of erythrocytes and liposomes by Electron Spin Resonance (ESR) using two spin labels 5-doxyl-stearate (5-NS) and 16-doxyl-stearate (16-NS) and by DPH (1,6-diphenyl 1,3,5-hexatriene) fluorescence polarization analysis. Lead induces decrease of fluidity in natural membrane which is more evident in membrane hydrophobic core; the effect is not correlated with lead concentration. Lead ion interacts with negative charged liposomes inducing an increase of their microviscosity. PMID- 6289852 TI - [Comparative evaluation of the effects of intravenous administration of cimetidine and ranitidine on several cardiovascular parameters]. AB - We have compared the effects of intravenous administration of cimetidine and ranetidine on some cardiovascular parameters. Five healthy volunteers received both cimetidine (3, 5 mg/Kg) and ranetidine (1, 5 mg/Kg). Heart rate, blood pressure and PEP/LVET were recorded at baseline and 5, 10, 30, 45 minutes after administration of both drugs. Intravenous administration of cimetidine and ranetidine did not induce any significant alterations in cardiovascular variables. PMID- 6289853 TI - [Immune response in the chicken after intrabursal administration of silica: preliminary observations]. AB - Immune response was studied in various experimental conditions using silica, a macrophage-toxic substance, in order to assess the role attributable to FAE cells, which have been demonstrated to be sensitive to this substance. Antibody response was tested in four different groups comprising seven animals each; the priming effect was studied in particular. It was observed that the silica considerably reduced the priming effect when the dose of antigen was administered in the bursal lumen at times immediately subsequent to the administration of the silica. The priming effect, on the contrary, was still present and was more marked when six hours elapsed between the administration of the silica and the priming dose. PMID- 6289854 TI - [Platelet anti-aggregant drugs and ischemic cardiopathy: evaluation of the effect on platelet aggregation and on antithrombin III]. PMID- 6289855 TI - [An autopsy case of Parkinson's disease associated clinically with dementia terminating in akinetic mutism and pathologically with multiple Lewy's Bodies in the cerebral cortex]. AB - An autopsy case of a 50 year-old male with Parkinson's disease associated with multiple Lewy bodies in the cerebral cortex was reported. His clinical symptoms began at the age of 26 with the speech and actions indicative of a persecution complex accompanied by irritability and were followed by progressive dementia from the age of 37 and Parkinsonism since the age of 41. He was in a state of akinetic mutism thereafter till his death at the age of 50. Autopsy disclosed in addition to the typical findings of Parkinson's disease in the brain stem multiple intracytoplasmic Lewy bodies in medium-sized neurons of the fifth and sixth layers of the cerebral cortex. They were atypical in the sense that they did not have any haloes. They were especially numerous in the cingulate gyrus. In addition, findings of non-specific neuronal degeneration were obtained in the cerebral cortex such as cellular atrophy with massive deposition of lipofuscin pigments, central chromatolysis, cell loss and cellular gliosis in the third, fifth and sixth layers. These neuronal findings were also prominent in the cingulate gyrus. Such senile changes as senile plaques or granulo-vacuolar degeneration were not found, although there were a few foci of neurofibrillary degeneration in the hippocampal gyrus. Histochemically and electron microscopically, no difference was observed in the constituents of Lewy bodies between the brain stem and the cerebral cortex. Such autopsy findings and a review of the literature indicate that the dementia in this case may be related not only to the presence of Lewy bodies but also to the above-described, non specific neuronal degeneration in the bilateral cingulate gyri and surrounding frontal gyri. The standpoint of regarding a Parkinson's disease with multiple Lewy bodies in the cerebral cortex as an independent disease entity was criticized. PMID- 6289856 TI - Influence of premedication on plasma ACTH and cortisol concentrations in children during adenoidectomy. AB - The endocrine response to stress, as reflected by the plasma concentrations of ACTH and cortisol, was investigated in 14 children receiving two different premedications during halothane anaesthesia for adenoidectomy. Seven children (group A) were premedicated with diazepam 5 mg rectally and atropine 0.3-0.4 mg sublingually and seven (group B) received a rectal combination of diazepam 0.5 mg kg-1, morphine 0.15 mg kg-1 and hyoscine 0.01 mg kg-1. Before and after surgery plasma concentrations of ACTH and cortisol were lower in group B than in group A. In group A mean values for ACTH increased from 40.7 ng line-1 before adenoidectomy to 352.9 ng litre-1 (P less than 0.001) after adenoidectomy. The corresponding increase in group B was from 12.1 ng litre-1 to 82.1 ng litre-1 (P less than 0.01). In group A mean cortisol concentrations increased from 235.7 nmol litre-1 to 655.7 nmol litre-1 after adenoidectomy (P less than 0.01) and in group B from 121.4 nmol litre-1 to 427.9 nmol litre-1 (P less than 0.01). End tidal carbon dioxide tension was approximately the same in both groups. It was concluded that the combination of diazepam, morphine and hyoscine decreased the endocrine response to stress. PMID- 6289857 TI - Pharmacodynamics of converting enzyme inhibition: the cardiovascular, endocrine and autonomic effects of MK421 (enalapril) and MK521. PMID- 6289858 TI - Enalapril maleate and a lysine analogue (MK-521): disposition in man. AB - 1 The disposition of two angiotensin converting-enzyme inhibitor drugs was studied in normal volunteers. One drug was enalapril maleate (MK-421), which requires in vivo esterolysis to yield active inhibitor (MK-422). The other was a lysine analogue of MK-422 (MK-521), which requires no bioactivation. 2 Absorption of enalapril maleate (10 mg, p.o.) was rapid, with peak serum concentrations of enalapril observed 0.5-1.5 h after administration. Based upon urinary recovery of total drug (enalapril plus MK-422), absorption was at least 61%. Bioactivation appeared to be largely post-absorptive. From the ratio of MK-422 to total drug in urine, the minimum extent of bioactivation was estimated at 0.7. 3 A similar dose of MK-521 was absorbed more slowly, reaching peak serum concentrations 6-8 h following drug administration. Minimum absorption, based upon urinary recovery, was 29%. 4 Serum concentration v time profiles for both drugs were polyphasic and exhibited prolonged terminal phases. 5 Recovery in urine and faeces of administered enalapril maleate (intact and as MK-422) was 94%. Recovery of MK-521 was 97%. These results indicate lack of significant metabolism of these agents, apart from the bioactivation of enalapril. PMID- 6289860 TI - The effects of fenfluramine on in vitro platelet aggregation. PMID- 6289859 TI - Enalapril maleate and a lysine analogue (MK-521) in normal volunteers; relationship between plasma drug levels and the renin angiotensin system. AB - 1 Two single doses of 10 mg each of the converting enzyme inhibitor enalapril maleate or MK-421 and of its lysine analogue (MK-521) were administered p.o. to twelve male volunteers. 2 The active diacid metabolite of MK-421 and the lysine analogue were determined by radioimmunoassay and MK-421 by the active metabolite method following in vitro hydrolysis. 3 Peak serum levels of MK-421, active metabolite and lysine analogue were reached within 1, 3 to 4, and 6 h respectively. Practically all MK-421 had disappeared from serum within 4 h. 4 A close correlation between percent inhibition of plasma converting enzyme activity and the serum concentration of active metabolite was observed ( r = 0.98, n = 171, P less than 0.001). Similarly, converting enzyme blockade as expressed by the ratio plasma angiotensin II/angiotensin I was closely correlated with serum active metabolite levels (r = 0.93, n = 15, P less than 0.001). PMID- 6289861 TI - Computed tomography of the thorax in the diagnosis and management of malignant disease. AB - The role of computed tomography (CT) scanning in the investigation of patients with intrathoracic malignancy has not yet been clearly defined. Current evidence suggests that it is the most sensitive diagnostic imaging technique for identifying the extent of primary lung cancers and their relationship to intrathoracic anatomy. It is an advance in the staging of intrathoracic malignancy as it identifies adenopathy in the paratracheal region and the mediastinum more readily. Discovery of small (less than 0.8 mm) pulmonary nodules is common in patients with some extrathoracic malignant conditions, but differentiating malignant deposits from benign nodules remains a problem. CT has a potential place in radiotherapy field planning and in identifying lung cancers in the occasional patient presenting with suspicious symptoms or positive cytology and a normal chest radiograph. PMID- 6289863 TI - The interaction of alpha thalassaemia with heterozygous beta thalassaemia. AB - The alpha globin genotypes of 55 beta thalassaemia heterozygotes have been determined by restriction endonuclease analysis to identify those with interacting alpha thalassaemia genes. A comparison of the haematological and haemoglobin synthesis findings of individuals with normal alpha genotypes (alpha alpha/alpha alpha) with those with one (-alpha/alpha alpha) or two (-alpha/ alpha) alpha genes deleted shows that the latter two groups have more balanced globin chain synthesis ratios, higher haemoglobin levels, and larger, better haemoglobinized red cells. This suggests that the degree of globin chain imbalance is a significant factor in determining the red cell characteristics in heterozygous beta thalassaemia. Screening programmes for thalassaemia, based on the detection of low MCVs, could miss cases of the interaction of alpha and beta thalassaemia. PMID- 6289864 TI - The molecular basis for beta o thalassaemia intermedia in an Iranian individual. AB - A symptomless Iranian patient homozygous for beta thalassaemia has haematological changes similar to the beta thalassaemia trait. This remarkably mild phenotype is probably the result of coexistent alpha thalassaemia and increased gamma chain synthesis. Restriction endonuclease mapping analysis of the beta globin genes indicates that the patient is homozygous for a single nucleotide substitution at the 5' donor splice junction in the second intervening sequence of the beta globin gene. No other changes were observed in the non-alpha globin gene cluster. It seems unlikely that the augmented gamma chain synthesis in this patient is related to the molecular defect responsible for this beta o thalassaemia. PMID- 6289865 TI - Androblastoma and raised serum alpha-fetoprotein with familial multinodular goitre. Case report. PMID- 6289862 TI - Pagetoid reticulosis--is it a disease entity? PMID- 6289866 TI - Cloned complementary deoxyribonucleic acid probes for untranslated messenger ribonucleic acid components of mouse sarcoma ascites cells. AB - Mouse sarcoma ascites cells contain several abundant mRNA species that occur to a large extent in an untranslated state. RNA preparations enriched in these species were used as starting material to construct recombinant plasmids. Cloned plasmids bearing sequences homologous to four of the untranslated mRNA species were identified by translation of hybrid-selected material. These plasmids, as well as a recombinant plasmid derived from chick alpha-actin mRNA, were used as probes for the estimation of mRNA levels in polyribosomes and in small ribonucleoprotein (RNP) particles of the ascites cells. Considerable amounts of the mRNA molecules belonging to the untranslated species were present in polyribosomes as well as in mRNPs. The actin mRNA, on the other hand, was present almost exclusively in polyribosomes. The distributions obtained by the hybridization assay resembled those estimated by translation of the same RNA preparations in cell-free systems. This indicates that the mRNA molecules of a given species engaged in translation in the cells and those present as untranslated RNP particles are equally effective in cell-free translation systems. PMID- 6289867 TI - Resolution of multiple heme centers of hydroxylamine oxidoreductase from Nitrosomonas. 1. Electron paramagnetic resonance spectroscopy. PMID- 6289868 TI - Proton nuclear magnetic resonance studies on cyclic nucleotide binding to the Escherichia coli adenosine cyclic 3',5'-phosphate receptor protein. PMID- 6289869 TI - Equilibrium binding of spin-labeled fatty acids to bovine serum albumin: suitability as surrogate ligands for natural fatty acids. AB - Electron paramagnetic resonance (EPR) and saturation transfer EPR (ST-EPR) spectroscopies were used to characterize the binding of spin-labeled fatty acid (SLFA) to bovine serum albumin (BSA). Association constants of three stearic acid derivatives labeled with a nitroxyl radical at C-5, C-12, or C-16 were estimated by EPR spectroscopy as the ratio of SLFA to BSA was increased from about 0 to 9. The values were compared to those for unmodified stearate. With all three SLFA, it was apparent that the nitroxyl residue modified the binding pattern. For SLFA:BSA ratios up to 1, which probably involves the site(s) on BSA most specific for long-chain FA, the C-16 derivative bound with an affinity similar to that of the natural FA. At higher ratios, the association constants for this SLFA were lower than those for stearate. The C-12 and C-5 derivatives showed only low affinity binding relative to stearate. The spectral parameter, W, was constant for SLFA:BSA ratios between 0 and 1 in the case of C-16 compound, indicating physical homogeneity of the high-affinity binding site. At higher ratios, the spectra changed progressively, indicating inhomogeneity of the lower affinity binding sites although parallel changes in association constants were not observed. Changes in W due to Heisenberg spin exchange were ruled out. By examining the mobility profile of the bound SLFA by both EPR and ST-EPR techniques, it was shown that the nitroxyl group was maximally immobilized when attached near the center of the carbon chain of the bound SLFA. PMID- 6289870 TI - Syntheses of biologically active ubiquinone derivatives. AB - Various 6-alkylubiquinone or 6-(omega-haloalkyl)ubiquinone derivatives were synthesized through a radical coupling reaction between alkanoyl or omega haloalkanoyl peroxides and ubiquinone 0. The latter was synthesized from 2 methoxy-4-methylphenol via nitration, methylation, reduction, and oxidation by modifications of the reported methods. 6-(omega-Haloalkyl)ubiquinones were converted to 6-(omega-hydroxyalkyl)ubiquinones by a mercuric-assisted solvolysis technique. The 6-(omega-hydroxyalkyl)ubiquinones were then esterified with carboxylic acid anhydrides or carboxylic acid bearing reporting groups, such as a photoaffinity label, N-(4-azido-2-nitrophenyl)-beta-alanine, or a spin-label, 3 carboxy-2,2,5,5-tetramethyl-3-pyrrolinyl-1-oxy. The esterification was catalyzed by dicyclohexylcarbodiimide and pyridine, and the esters were purified by preparative silica gel thin-layer chromatography, developed by 3% ethanol in benzene. The spectral properties and biological functions of the synthesized ubiquinone derivatives were studied. The biological function of the synthesized compounds was followed by the ability to serve as an electron acceptor, donor, or mediator in the isolated mitochondrial electron transfer complexes of succinate-Q reductase, ubiquinol-cytochrome c reductase, and succinate-cytochrome c reductase, respectively. The concentration effect of these ubiquinone derivatives on the electron transfer reaction was compared with that of ubiquinone 10. The study of the inhibitory effect of synthesized arylazidoubiquinone on succinate cytochrome c reductase after photolysis confirmed the existence of specific Q binding proteins in this segment of the respiratory chain. The specific interaction between ubiquinone and protein has also gained support from the immobilization of the spin-label of a synthesized spin-labeled ubiquinone derivative. PMID- 6289871 TI - Iodination catalyzed by the xanthine oxidase system: role of hydroxyl radicals. PMID- 6289872 TI - Reconstitution of the transferrin receptor in lipid vesicles. Effect of cholesterol on the binding of transferrin. AB - Purified rabbit reticulocyte transferrin receptors were incorporated into phosphatidylcholine vesicles containing varying amounts of cholesterol. The binding of transferrin to the receptor in the reconstituted vesicles had three distinct characteristics: (1) The binding of transferrin exhibited the two components characteristic of transferrin binding to erythroid cells, a saturable, specific component and a nonsaturable, nonspecific component. (2) Transferrin binding exhibited positive cooperativity at low cholesterol/phospholipid (C/P) molar ratios. However, the cooperativity diminished and then disappeared as the C/P molar ratios were increased to the levels found in circulating red blood cells. (3) The amount of specific transferrin binding to the reconstituted vesicles also decreased as the C/P molar ratio was increased. These results indicate that in the reconstituted system the lipid environment plays a significant role in the expression of transferrin receptors. PMID- 6289873 TI - Two pH optima of adenosine 5'-triphosphate dependent deoxyribonuclease from Bacillus laterosporus. AB - The various catalytic activities of the ATP-dependent deoxyribonuclease (DNase) of Bacillus laterosporus have pH optima at 6.3 and 8.3. Although the pH profile of ATP-dependent DNase activity on duplex DNA is bell shaped with a maximum at about pH 8.3, ATP-dependent DNAse activity on single-stranded DNA has optima at pH 6.3 and 8.3. ATPase activities dependent on double-stranded and single stranded DNA have a high bell-shaped peak with a maximum at pH 6.3 with a low and broad shoulder at about pH 8.3. ATP-independent DNase activity also has optima at pH 6.3 and 8.3. The ratio of the amount of ATP hydrolyzed per number of cleaved phosphodiester bonds in DNA increases with decrease in the pH value of the reaction. The ratios obtained at pH 8.3 and 6.3 were respectively about 3 and 22 with duplex DNA as substrate and 5 and 17 with single-stranded DNA as substrate. Formation of a single-stranded region of 15000-20000 nucleotides, which is linked to duplex DNA and about half of which has 3'-hydroxyl termini, was observed at about pH 6.3, but not at above pH 7.5. Furthermore, the optimum concentrations of divalent cations for the activity producing the single-stranded region and the activity hydrolyzing ATP were identical (3 mM Mn2+ or 5 mM Mg2+). Thus the two activities are closely related. These results indicate that the enzyme has two different modes of action on duplex DNA which are modulated by the pH. PMID- 6289874 TI - Studies on the methylene/methyl interconversion catalyzed by methylenetetrahydrofolate reductase from pig liver. PMID- 6289875 TI - Oxidative decarboxylation of benzoate to carbon dioxide by rat liver microsomes: a probe for oxygen radical production during microsomal electron transfer. PMID- 6289876 TI - Evidence on the existence of a purine ligand induced conformational change in the active site of bovine pancreatic ribonuclease A studied by proton nuclear magnetic resonance spectroscopy. AB - The titration curves of the C-2 histidine protons of RNase A and of derivative II -a covalent derivative obtained by reaction of the enzyme with the halogenated nucleotide 9-beta-D-ribofuranosyl-6-chloropurine 5'-phosphate--in the presence of a number of purine nucleosides, nucleoside monophosphates, and nucleoside diphosphates were studied by means of proton nuclear magnetic resonance at 270 MHz. The examination of the perturbations found on the chemical shifts and pKs of the C-2 protons of His-12, -48, and -119 are consistent with the following conclusions: (1) The interaction of adenosine in the primary purine binding site of the enzyme (B2R2) induces a conformational change in the active center of the enzyme [for the nomenclature of the RNase A binding subsites, see Pares et al. [Pares, X., Llorens, R., Arus, C., & Cuchillo, C. M. (1980) Eur. J. Biochem. 105, 571-579]]. (2) The phosphate moiety of the ligands, independently of its position, probably acts as a general carrier of the nucleotide to the active center, while the substituents of the base are the generators of the specificity of the binding and control the binding equilibrium between subsites B2R2 and B1R1. (3) There is no overlapping between the binding sites occupied by the labeling nucleotide in derivative II (B3R3p2) and the primary binding site for purine mononucleotides (B2R2p1). PMID- 6289877 TI - Identification and partial purification of a factor that stimulates calcium dependent proteases. PMID- 6289878 TI - Nitrocellulose filter binding studies of the interactions of Escherichia coli RNA polymerase holoenzyme with deoxyribonucleic acid restriction fragments: evidence for multiple classes of nonpromoter interactions, some of which display promoter like properties. PMID- 6289879 TI - Poly(7-deazaguanylic acid), the homopolynucleotide of the parent nucleoside of queuosine. AB - Poly(7-deazaguanylic acid) was enzymatically synthesized by the polymerization of 7-deazaguanosine 5'-diphosphate with polynucleotide phosphorylase from Micrococcus luteus in high yield. The homopolymer showed a similar thermal and total hypochromicity to poly(G) at the long wavelength absorption maximum. No sigmoid melting profile was observed for poly(c7G) as is found for poly(G), implying a single-stranded structure in aqueous solution. From the circular dichroism spectra it can be concluded that the 7-deazapurine nucleotide is much more flexible than the purine nucleotide. In analogy to poly(G), the homopolymer poly(c7G) forms a 1:1 complex with poly(C) under neutral conditions, melting at a similar temperature to the poly(G) complex. However, at pH 2.5, where a poly(G) X 2poly(C) complex is observed, poly(c7G) still binds only one poly(C) strand. This is due to the lack of N-7 in poly(c7G), not allowing Hoogsteen base pair formation, which occurs with poly(G). RNase T1 cleaves poly(c7G), indicating that N-7 of guanosine is not a requirement for nucleotide binding to the enzyme, as has been suggested. Because of the single-stranded structure of poly(c7G), the polynucleotide chain is rapidly hydrolyzed by the single-strand-specific nuclease S1, whereas multistranded poly(G) is completely resistant. PMID- 6289880 TI - A kinetic study of interactions of (Rp)- and (Sp)-adenosine cyclic 3',5' phosphorothioates with type II bovine cardiac muscle adenosine cyclic 3',5' phosphate dependent protein kinase. AB - The stereoselectivity of the adenosine cyclic 3',5'-phosphate (cAMP) binding sites on the regulatory subunit of the type II bovine cardiac muscle cAMP dependent protein kinase was investigated by examining the interactions of (Rp)- and (Sp)-adenosine cyclic 3',5'-phosphorothioates (cAMPS) with these sites. While activation of the holoenzyme and binding to the regulatory subunit of the type II kinase were observed for both of these diastereomers, there were significant differences between the interactions of the cAMPS isomers with the enzyme. In particular, the Sp isomer is more potent than the Rp species not only in the activation of reconstituted, as well as directly isolated, holoenzyme but also in the inhibition of [3H]cAMP binding to the regulatory subunit. A marked preference for the binding of the Sp isomer to site 2 in the regulatory subunit exists. Hydrogen bonding of a functional group on the regulatory subunit with preferential orientation toward the exocyclic oxygen rather than the sulfur of the thiophosphoryl residue may be involved in the observed selectivity of cAMPS binding and activation. In addition to our findings on the stereoselectivity of the binding of cAMPS to cAMP-dependent protein kinase, we have established a method for the reconstitution of holoenzyme from the purified subunits without subjecting the regulatory protein to denaturing conditions. PMID- 6289881 TI - Proton release during the pre-steady-state oxidation of aldehydes by aldehyde dehydrogenase. Evidence for a rate-limiting conformational change. AB - A transient release of protons with an amplitude corresponding to one proton per active site has been observed for the oxidation of propionaldehyde, acetaldehyde, and benzaldehyde by sheep liver cytoplasmic aldehyde dehydrogenase at pH 7.6 with phenol red as indicator. At saturating substrate levels, the rate constants for the proton burst are in each case the same, and for acetaldehyde and propionaldehyde show the same dependence on the concentrations of the substrates, as the rate constants for the transient production of NADH reported previously [MacGibbon, A.K.H., Blackwell, L.F., & Buckley, P.D. (1977) Biochem. J. 167, 469 477]. Although, with propionaldehyde as a substrate, a full proton burst is also observed at pH 6.0, no proton burst is observed at pH 9.0. For 4 nitrobenzaldehyde, there is no burst in NADH production, but a burst in proton release is observed, showing that proton release precedes hydride transfer. No protons were released during the binding of the substrate analogues acetone and chloral hydrate nor on reaction of the enzyme with the inhibitor tetraethylthiuram disulfide (disulfiram). A model is proposed in which the rate limiting step in the pre-steady-state phase of the reaction is a conformational change which occurs after the binding of aldehydes to the enzyme. As a result of the conformational change, the environment of a functional group on the enzyme, which initially has a pKa of about 8.5, is perturbed to give a final pKa value for the group of less than 5. Computer simulations were used to show that the model accurately reproduces all of the experimental data. The lack of observation of a second transient proton release, as required by the overall stoichiometry, argues that its release occurs in a slow step prior to NADH dissociation. PMID- 6289882 TI - Oxidation of indole-3-acetic acid by peroxidase: involvement of reduced peroxidase and compound III with superoxide as a product. AB - Kinetic and spectral data establish that peroxidase may oxidize indole-3-acetic acid by either of two pathways depending on the enzyme/substrate ratio. When relatively low enzyme/substrate ratios are employed, the oxidation proceeds through a reduced peroxidase in equilibrium compound III shuttle. Conversely, peroxidase operates through the conventionally accepted pathway involving native enzyme and compounds I and II only when high enzyme/substrate ratios are used. Compound III, a specific oxidase, constitutes the dominant steady-state form of peroxidase when the reduced peroxidase in equilibrium compound III shuttle is operational. Activation of this shuttle also produces a flux of superoxide anion radical at the expense of molecular oxygen. Thus, important biological consequences may follow activation of this shuttle under physiological conditions. PMID- 6289884 TI - Proton stoichiometry of adenosine 5'-triphosphate synthesis in rat liver mitochondria studied by phosphorus-31 nuclear magnetic resonance. PMID- 6289883 TI - Models for slow anisotropic rotational diffusion in saturation transfer electron paramagnetic resonance at 9 and 35 GHz. AB - Model systems of cholestane and 5-doxylstearic acid analogue spin probes in lipid bilayer dispersions of dipalmitoylphosphatidylcholine and cholesterol (9:1 w/w) are used to analyze saturation transfer electron paramagnetic resonance spectral behavior for slow rotational diffusion in an anisotropic medium. Measurements are made at both 9 and 35 GHz to provide enhanced spectral resolution for different types of motion. Parameter correlation plots of spectral parameters from different regions of the saturation transfer spectra appear to be potentially useful in characterizing different types of motion. Anisotropic rotational diffusion about a symmetry axis coincident with the nitroxide y principal axis is clearly distinguishable from isotropic rotational diffusion and may be distinguishable from rotational diffusion about the nitroxide z principal axis. Approximate anisotropic rotational diffusion about a symmetry axis coincident with the nitroxide z principal axis is distinguishable from isotropic rotational diffusion under some, but not all, conditions. PMID- 6289885 TI - Organization of herpes simplex virus type 1 deoxyribonucleic acid during replication probed in living cells with 4,5',8-trimethylpsoralen. AB - The structure of herpes simplex virus type 1 (HSV-1) DNA in the nuclei of living infected cells was studied with the DNA photoaffinity probe 4,5',8 trimethylpsoralen. The rate of photobinding to HSV-1 DNA was compared to that of a suitable internal control at different times during infection. The rates of photobinding to DNA packaged in virions, capsids, and prereplicative and postreplicative DNA were characteristically different. By 4 h after infection, after the initiation of DNA replication, the rate of photobinding to HSV-1 DNA increased 4 times relative to the rate of binding to the host DNA. The enhanced rate of photobinding to HSV-1 DNA was maintained at all later times during infection and was not affected when frequent single-strand breaks were introduced in HSV-1 DNA by gamma irradiation of infected cells. The results suggest that the bulk of the replicating herpes DNA is free of torsional tension and that the differing rates of photobinding are attributable to changes in accessibility of the HSV-1 DNA. The results are compatible with previous proposals, based on in vitro studies, that intranuclear HSV-1 DNA is primarily free of nucleosomal organization and suggest that there are few, if any, unrestrained DNA supercoils averaged over the entire HSV-1 genome. PMID- 6289887 TI - Mitochondrial lipid peroxidation by cumene hydroperoxide and its prevention by succinate. AB - Rat liver mitochondria form lipid hydroperoxides when they are incubated aerobically with cumene hydroperoxide. The rate of reaction is dependent on the initial concentration of the latter and involves the consumption of oxygen. Gradient-separated and cytochrome c-depleted mitochondria, mitoplasts and submitochondrial fractions also undergo this peroxidation. Mitochondrial lipid peroxidation by cumene hydroperoxide is strongly inhibited by SKF52A (an inhibitor of cytochrome P-450), by antioxidants and to a lesser extent by the enzymes superoxide dismutase and catalase. Conversely, rotenone and N ethylmaleimide stimulate the reaction. Succinate protects against the lipid peroxidation and in some mitochondrial fractions the associated oxygen uptake is also inhibited. This protection by succinate is prevented by malonate but not by N-ethylmaleimide or antimycin. Lipid hydroperoxides present in previously peroxidised mitochondria are partly lost on reincubation with succinate and this reaction is also unaffected by N-ethylmaleimide but inhibited by both malonate and antimycin. The results suggest that reduction of mitochondrial ubiquinone may prevent the generation of lipid hydroperoxides but that their subsequent removal may require reduction at or beyond cytochrome b. PMID- 6289886 TI - Purification and characterization of highly purified cytochrome b from complex III of baker's yeast. AB - A simple, high-yield purification procedure for cytochrome b from yeast Complex III has been developed. This procedure involves solubilization using chemical modification of the lysine residues with 3,4,5,6-tetrahydrophthalic anhydride followed by hydroxyapatite column chromatography. This purified cytochrome b has a heme content of 37.0 nmol cytochrome b/mg and a molecular weight on SDS gels of 25000-26000. Amino acid analysis indicates high hydrophobicity and is very comparable to the composition deduced from the gene sequence (Nobrega, F.G. and Tzagoloff, A. (1980) J. Biol. Chem. 255, 9828-9837). The latter data indicate a molecular weight of 42000 for the polypeptide; our heme analyses thus imply the presence of two hemes per polypeptide chain. Optical and MCD spectra are typical of a low-spin b-type cytochrome. MCD-potentiometric titration indicates a one electron carrier with a single midpoint potential of -44 mV at pH 7.4 and 25 degrees C. The EPR spectrum of isolated cytochrome b has only one gz signal at 3.70, indicating that the 'strained' heme structure (Carter, K., T'sai, A. and Palmer, G. (1981) FEBS Lett. 132, 243--246) is still maintained. No indication of antimycin binding was demonstrated either by the direct-fluorescence method or binding-precipitation method although stoichiometric binding to the parent Complex III was readily demonstrated. PMID- 6289888 TI - The role of the sites for ATP of the Ca2+ -ATPase from human red cell membranes during Ca2+ -phosphatase activity. PMID- 6289889 TI - Characterization of the the (Na + +K+)-ATPase in a membranous preparation from the optic ganglion of the squid (Loligo pealei). AB - (1) A membrane fraction enriched in (Na+ +K+)-ATPase (EC 3.6.1.3) was obtained from optic ganglia of the squid (Loligo pealei) by density gradient fractionation of membranes followed by treatment with either SDS or Brij-58. The resulting membrane had an (Na+ +K+)-ATPase specific activity of approx. 2 units/mg and was greater than 95% ouabain-sensitive. (2) The (Na+ +K+)-ATPase had a Km for ATP of 0.42 +/- 0.04 mM and a pH optimum of 7.0. It was inhibited by ouabain with a Ki of 0.32 +/- 0.04 microM. (3) Optimum monovalent cation concentrations were: 240 mM NaCl, 60 mM KCl, tested with NaCl + KCl = 300 mM. (4) The Mg2+ dependence of hydrolysis varied with the absolute ATP concentration. At 3 mM ATP, the Km for Mg2+ was 0.86 +/- 0.10 mM, and at 6 mM ATP, the Km was 1.86 +/- 0.44 mM. High levels of Mg2+ caused inhibition of hydrolysis. (5) The interactions of Na+ and K+ were examined over a range of conditions. K+ levels caused modulations in the Na+ dependence in the range of 1-150 mM. (6) The (Na+ +K+)-ATPase prepared from squid optic ganglion displays properties similar to those of the sodium pump in injected nerves. PMID- 6289890 TI - Phospholipid and enzyme arrangements of rat liver rough microsomal subfractions from control and methylcholanthrene-treated animals. AB - Rough microsomes from rat liver of both control and methylcholanthrene-treated animals were subfractionated on a discontinuous sucrose gradient into three fractions according the their sedimentation velocity. The slowly sedimenting vesicles were enriched in electron transport enzymes, while those in the pellet showed higher phosphatase and ATPase activities. Methylcholanthrene treatment introduced typical changes in enzyme composition, mainly an increase of the cytochrome P-448. The individual phospholipids exhibited an identical distribution pattern in the three subfractions and no change occurred after induction with methylcholanthrene treatment. Nearest neighbour analysis of phosphatidylethanolamine with dinitrodifluorobenzene revealed a similar pattern in the enzymatically different subfraction, that is, no cross-linking with phosphatidylserine occurred. One-third of the phosphatidylethanolamine was in monomer and dimer form and about two-thirds was protein linked. When membrane and enzyme synthesis was induced, cross-linking to proteins were substantially decreased. The experiments indicate that the phospholipids are distributed in a homogenous fashion in the lateral plane of the rough microsomal membrane and do not support the possibility that phosphatidylethanolamine is specifically associated with cytochrome P-450. PMID- 6289892 TI - Polycation-induced fusion of negatively-charged vesicles. AB - Sonicated vesicles of 20-50 nm in diameter consisting of neutral phospholipids and a variety of acidic phospholipids were interacted with polylysine, cytochrome c, Ca2+ and Mg2+. The addition of polycations caused massive aggregation accompanied by an increase of membrane permeability as determined by leakage of fluorescent dye. Aggregation was followed by fusion of the vesicles into structures that in some cases exceeded 1 micron in diameter. Polylysine induced aggregation and appreciable fusion at charge ratios (polylysine/phospholipid) of 0.5-2, while divalent cations did so only at charge ratios (cation/phospholipid) greater than 10. Aggregation and fusion induced by polylysine were dependent also on the size of the polycation, i.e., the longer the molecule the less needed to induce similar aggregation. It appears that, due to the concentration of charges on a single molecule, polylysine is at least an order of magnitude more effective than divalent cations at inducing fusion of membranes. Cytochrome c induced fusion of similar vesicles at moderately acidic pH (pH 4.2). PMID- 6289891 TI - Crystallization patterns of membrane-bound (Na+ +K+)-ATPase. AB - Extensive formation of two-dimensional crystals of the proteins of the pure membrane-bound (Na+ +K+)-ATPase is induced during prolonged incubation with vanadate and magnesium. Some membrane crystals are formed in medium containing magnesium and phosphate. Computer-averaged images of the two-dimensional crystals show that the unit cell in vanadate-induced crystals contains a protomeric alpha beta-unit of the enzyme protein. In phosphate-induced crystals an (alpha beta) 2 unit occupies one unit cell suggesting the interactions between alpha beta-units can be of importance in the function of the Na+, K+ pump. PMID- 6289893 TI - Guanidine hydrochloride-induced shedding of a Dictyostelium discoideum plasma membrane fraction enriched in the cyclic adenosine 3',5'-monophosphate receptor. AB - The cell surface cyclic AMP receptor of Dictyostelium discoideum is under study in a number of laboratories with respect to both its role in development of the organism and the physiology of excitation-response coupling. We report here that when starved amoebae are exposed to the chaotrope guanidine hydrochloride at 1.8 M, they shed a particulate cyclic AMP binding activity into the medium. This activity is due to membrane vesicles which originate from the cell surface. The vesicles are enriched up to 150-fold in cyclic AMP binding activity and up to 14 fold in phospholipid content when compared to the starting amoebae. The cyclic AMP binding activity of the membrane vesicles is identical to that of the cell surface receptor with respect to the following properties; (i) it is lacking in preparations from unstarved, vegetative amoebae; (ii) it is not inhibited by cyclic GMP and is stimulated by calcium ions; (iii) it has very rapid rates of association and dissociation of bound cyclic AMP; (iv) it has two classes of binding sites with dissociation constants similar to those of the surface receptors of whole amoebae. The binding activity of the isolated membranes is stable for several days at 4 degrees C and the lower affinity binding sites are stable up to several months when stored at -80 degrees C. Due to enrichment and stability of the receptor in this preparation, it should be highly suitable for many types of studies. The usefulness is enhanced by the fact that the preparation does not contain detectable cyclic AMP phosphodiesterase activity. PMID- 6289894 TI - K+ transport in 'tight' epithelial monolayers of MDCK cells. Evidence for a calcium-activated K+ channel. AB - Measurements of 86Rb efflux across the apical and basal-lateral aspects of intact monolayers of 'high-resistance' MDCK cells mounted in Ussing chambers have been made. A transient increase in 86Rb efflux across both epithelial borders upon stimulation with adrenalineeeeeee or ionophore A23187 is observed. The increased 86Rb across the basal cell aspects is of greatest quantitative importance. Measurements of total cellular K+ contents by flame photometry of tissue extracts indicate a net loss of K+ following adrenalin addition. The effects of adrenalin and ionophore A23187 upon 86Rb efflux are abolished in 'Ca2+ -free' media. The properties of the Ca2+ -dependent increase in 86Rb efflux show similarities to Ca2+ -activated K+ conductances in other tissues, notably human red cells, including inhibition by quinine (1 mM), tetraethylammonium (25 mM) and insensitivity to bee venom toxin (apamin) (25 nM). Adrenalin is only effective when applied to the basal bathing solution suggesting that the receptors mediating adrenalin action are located upon the basal-lateral membranes. Half maximal stimulation of 86Rb efflux by adrenalin is observed at 9.1 X 10(-7) M. The action of various adrenergic receptor agonists and antagonists are consistent with adrenalin action being mediated by an alpha-adrenergic receptor. PMID- 6289895 TI - Studies on (K+ + H+)-ATPase V. Chemical composition and molecular weight of the catalytic subunit. AB - (1) A (K+ + H+)-ATPase preparation from porcine gastric mucosa is solubilized in sodium dodecyl sulfate, and is subjected to gel filtration. (2) A main subunit fraction is obtained, which is a protein carbohydrate lipid complex, containing 88% protein, 7% carbohydrate and 5% phospholipid. The Detailed composition of the protein and carbohydrate moieties are reported. (3) Sedimentation analysis of the subunit preparation, after detergent removal, reveals no heterogeneity, but the subunits readily undergo aggregation. (4) Acylation of the subunit preparation with citraconic anhydride causes a clear shift of the band obtained after SDS gel electrophoresis, but the absence of broadening and splitting of the band pleads against subunit heterogeneity. (5) Treatment of the subunit preparation with dansyl chloride indicates that the NH2 terminus is blocked, which favors the assumption of homogeneity of the protein. (6) Binding studies with concanavalin A indicate that at least 86% of the subunit preparation is composed of glycoprotein. (7) These findings, taken together, strongly suggest that there is a single subunit which is a glycoprotein and which represents the catalytic subunit of the enzyme. From sedimentation equilibrium analysis a molecular mass value of 119 kDa (S.E. 3, n = 6) is calculated for protein + carbohydrate and of 110 kDa (S.E. 3, N = 6) for protein only. (8) In combination with the molecular mass of 444 kDa (S. E. 10, n = 4) obtained for the intact enzyme by radiation inactivation we conclude that the enzyme appears to be composed of a homo tetramer of catalytic subunits. PMID- 6289896 TI - Use of percollTM in the isolation and purification of rabbit small intestinal brush border membranes. AB - (1) Intestinal absorption is altered under a variety of circumstances in health and disease and to determine a possible relationship between intestinal absorptive function and intestinal brush border membrane composition, we undertook the isolation and purification of rabbit jejunal and ileal brush borders, to allow further studies of their lipid composition under varied experimental conditions. (2) A modification of an established method (Schmitz, J., Preiser, H., Maestracci, D., Ghosh, B.K., Cerda, J.J. and Crane, R.K. (1973) Biochim. Biophys. Acta 323, 98-112) utilized CaCl2 aggregation and sequential centrifugation followed by purification of the brush border pellet (P2) at 27,000 X g on a PercollTM (Pharmacia) self-forming gradient. The PercollTM was removed by ultracentrifugation for 30 min at 100 000 X g, utilizing a batch rotor in the Beckman airfugeTM. (3) Pure brush border membrane vesicles were obtained and characterized by specific marker analysis and electron microscopy. Comparative marker analyses performed on P2 and final PercollTM preparations from animals showed that the purification achieved was 8-11-fold greater when compared to the original homogenates. Verification of purity was also demonstrated by the absence of DNA and very low levels of Beta-gluconridase and (Na+ + K+)-ATPase in the PercollTM preparations. (4) Comparative lipid analyses of P2 and final PercollTM preparations showed that levels of total phospholipid and free fatty acids were several-fold higher in the PercollTM preparations on a per mg protein basis. (5) A comparison of the activity of enzyme markers and the levels of total free fatty acids in P2 pellets obtained after Cacl2 and MgCl2 aggregation showed that CaCl2 aggregation gave the more consistently reproducible results. (6) Although standard procedures of membrane preparations not involving density gradient separation provide membranes of reasonable purity for the estimation of lipid components, we consider the final purification step of density gradient separation using PercollTM is essential for determining small quantitative changes which might occur in the membrane lipid composition under experimental conditions were intestinal absorptive function is altered. PMID- 6289897 TI - PhoE protein pore of the outer membrane of Escherichia coli K12 is a particularly efficient channel for organic and inorganic phosphate. AB - This study was undertaken to investigate the proposed in vivo pore function of PhoE protein, an Escherichia coli K12 outer membrane protein induced by growth under phosphate limitation and to compare it with those of the constitutive pore proteins OmpF and OmpC. Appropriate mutant strains were constructed containing only one of the proteins PhoE, OmpF or OmpC, or none of these proteins at all. By measuring rates of nutrient uptake at low solute concentrations, the proposed pore function of PhoE protein was confirmed as the presence of the protein facilitates the diffusion of Pi through the outer membrane, such as a pore protein deficient strain behaves as a Km mutant. Comparison of the rates of permeation of Pi, glycerol 3-phosphate and glucose 6-phosphate through pores formed by PhoE, OmpF and OmpC proteins shows that PhoE protein is the most effective pore in facilitating the diffusion of Pi and phosphorus-containing compounds. The three types of pores were about equally effective in facilitating the permeation of glucose and arsenate. Possible reasons for the preference for Pi and Pi-containing solutes are discussed. PMID- 6289898 TI - Mechanism of the Na+, K+ pump. Protein structure and conformations of the pure (Na+ +K+)-ATPase. PMID- 6289899 TI - Cloning in a cosmid vector of complete 37 kb and 25 kb ribosomal DNA repeat units from the chicken. AB - DNA fragments of up to 40 kb containing rRNA-coding sequences have been isolated from a chicken liver DNA library prepared in the cosmid pHC79. Characterization of the cloned DNA by R-loop and restriction mapping has shown that there are two size classes of repeat unit, one of 37 kb and one of 25 kb, the larger of which is a family of units which vary slightly in size. These two classes were shown to be present in the DNA of a single chicken. The size of the internal transcribed spacer in the chicken was measured to be 4.4 kb from analysis of R-loops and heteroduplexes between chicken and Xenopus laevis rDNAs. No introns were observed in either the 18 S or the 28 S coding sequences. The number of copies of the chicken rDNA unit was measured by titration against the cloned sequences to be 202 +/- 51 per haploid genome. PMID- 6289900 TI - Phosphorylation of ribosomal proteins during the cell cycle of Physarum polycephalum. AB - Physarum polycephalum has been used as a model system to study the phosphorylation of ribosomal proteins during the cell cycle. The results showed that the phosphate content of S3, the major ribosomal phosphoprotein in this organism, was constant during all phases of the cell cycle. No additional ribosomal phosphoproteins were observed. These results differ significantly from those reported earlier by Rupp, R.G., Humphrey, R.M. and Shaeffer, J.R. (Biochim. Biophys. Acta (1976) 418, 81-92) and suggest that the use of thymidine or hydroxyurea to synchronize cell population may affect the phosphorylation of ribosomal proteins. The results are discussed in relation to protein synthesis and cAMP level during the cell cycle. PMID- 6289901 TI - The activity of pulmonary indoleamine 2,3-dioxygenase in rats and mice is not altered by oxygen exposure. AB - We tested the hypothesis that pulmonary indoleamine 2,3-dioxygenase (indole:oxygen 2,3-oxidoreductase (decyclizing), EC 1.13.11.17), an enzyme that consumes superoxide anion (O-2), might have an antioxidant role under conditions of hyperoxia. We measured indoleamine 2,3-dioxygenase in three experimental models in which pulmonary superoxide dismutase, catalase and glutathione peroxidase (the known antioxidant enzymes) show increased activity and are associated with greater tolerance to 96-98% O2 exposure: (1) adult rats preexposed to 85% O2 for 5-7 days; (2) neonatal rats exposed directly to greater than 95% O2; and (3) adult rats treated with bacterial endotoxin during O2 exposure. Indoleamine 2,3-dioxygenase did not increase in response to O2 exposure in any of these rat models. Conversely, in adult mice treated with endotoxin, lung indoleamine 2,3-dioxygenase activity did increase, but no protection against O2 toxicity occurred. Thus, a rise in indoleamine 2,3-dioxygenase is neither necessary nor sufficient to confer resistance to O2 toxicity. These data taken together are evidence against its having any important role in the antioxidant defense system of the lung. PMID- 6289902 TI - Identification and kinetic studies of an inducible mannokinase from a Streptomyces strain. AB - Crude extracts from cells of a Streptomyces strain isolated from a palm-grove soil and grown on different carbon sources showed a constitutive glucokinase. Specifically inducible kinase activity for mannose was found in cells grown on mannose or beta-D-mannan. The activity on mannose was due to a highly specific mannokinase (ATP:D-mannose 6-phosphotransferase, EC 2.7.1.7) which has been separated from the glucokinase by Ultrogel AcA 54 gel filtration chromatography. Initial velocity and inhibition product studies were carried out to investigate the reaction pathway. In the absence of products, reciprocal plots intersecting on the abscissas were observed when either mannose or ATP concentration was varied in the presence of several fixed concentrations of the non-varied substrate. Km values for mannose and Mg-ATP complex are 0.33 and 1.1 mM, respectively. In the product inhibition studies, mannose-6-P was observed to be competitive with mannose and non-competitive with Mg-ATP. The reverse was observed when ADP was used as inhibitor. These data are consistent with a random sequential Bi-Bi mechanism with two dead-end ternary complexes. PMID- 6289903 TI - Carboxypeptidase Y from Saccharomyces cerevisiae. Conformational differences reflected in kinetic behaviour in water and deuterium oxide. AB - The glycoenzyme carboxypeptidase Y (peptidyl--amino-acid hydrolase, EC 3.4.16.1), from baker's yeast (British Fermentation Products Strain, Ng 72), of molecular weight 60 000, had a protein portion closely similar to those in the literature for carboxypeptidase Y isolations from other yeast sources, but was 25.3 wt% carbohydrate (mannose 83.3% by wt., glucosamine 10.3% by wt. with traces of galactose and galactosamine). Circular dichroic spectra indicated that the enzyme lost its beta-structure as the pH was lowered from 8.08 to 4.16. At p2H 8.22 in 2H2O media the conformation of this enzyme was different from that observed at pH 8.08. A tyrosine residue appeared to be perturbed by lowering the pH of the medium. Carboxypeptidase Y was rapidly, and essentially irreversibly, inactivated at low p2H. The pH profile of kcat for the carboxypeptidase Y-catalysed hydrolysis of 4-nitrophenyltrimethylacetate showed two inflections at 45 degrees C: one at pKapp approximately 3.7 insensitive to temperature variation (ascribed to a carboxyl group), and one of pKapp approximately 5.7 markedly temperature dependent and possibly caused by a histidine residue. PMID- 6289904 TI - Estimation of molecular weights of membrane proteins in the presence of SDS by low-angle laser light scattering combined with high-performance porous silica gel chromatography. Confirmation of the trimer structure of porin of the E. coli outer membrane. AB - An assessment study was carried out to evaluate the performance of the low-angle laser light-scattering technique combined with high-performance porous silica gel chromatography in the presence of sodium dodecyl sulfate and precision differential refractometry. It was found that the combined technique is highly promising as a reliable method for determining the molecular weight of a membrane protein solubilized by the surfactant. As a test, molecular weights of porin forming the permeability channel of the outer membrane of E. coli B in an oligomeric form were measured before and after heat treatment, which is known to cause dissociation. The results obtained indicate that the porin oligomer is a trimer with stoichiometric composition. PMID- 6289906 TI - The demonstration in rat liver cell sap of protein kinase and phosphoprotein phosphatase active on fructose-bisphosphatase. AB - A protein kinase active on fructose-bisphosphatase (D-fructose-1,6-bisphosphate 1 phosphohydrolase, EC 3.1.3.11) was demonstrated in rat liver cell sap. The protein kinase activity was stimulated by cyclic AMP and coincided with the activity of cyclic AMP-dependent protein kinase type I. In addition, three different peaks of phosphoprotein phosphatase active on [32P] phosphofructose bisphosphatase were found on chromatography of rat liver cell sap on a DEAE cellulose column. These phosphatases needed divalent cations for full activity. 5'-AMP, a negative modulator of fructose-bisphosphatase, had no effect on the phosphorylation-de-phosphorylation reactions of the enzyme. ATP and Ca2+ did not influence the dephosphorylation reaction of fructose-bisphosphatase. PMID- 6289905 TI - The role of the nicotinamide moiety of NAd+ for negative cooperativity in glyceraldehyde-3-phosphate dehydrogenase as studied by spin-labeled cofactors. AB - Two derivatives of NAD+ spin-labeled at N6 or C-8 of the adenine ring have been shown previously to be active coenzymes of glyceraldehyde-3-phosphate dehydrogenase (D-glyceraldehyde-3-phosphate: NAD+ oxidoreductase (phosphorylating), EC 1.2.1.12). When more than two equivalents of either spin labeled NAD+ are bound to the tetrameric enzyme, spin-spin interaction is observed in the ESR spectra (Deparade, M.P., Gloggler, K. and Trommer, W.E. (1981) Biochim. Biophys, Acta 659, 422-433). After reduction of enzyme-bound NAD+ spin-labeled at C-8 to the corresponding NADH derivative, the additional peaks due to this spin-spin interaction disappear, which implies that the distance between the two radicals increases. It is proposed that the coenzyme slide further towards the active site upon reduction. ADPR spin-labeled at C-8 binds non-cooperatively, exhibiting a dissociation constant of Kd = 33 microM. Even with 3.5 equivalents bound to the enzyme, spin-spin interaction is not observed. AMP spin-labeled at C-8 combines with two sites per monomer, or a total of eight per tetramer. The respective dissociation constants are Kd1 = 30 microM and Kd2 - 2.3 mM. Phosphate competes with AMP bound to the weak site. Spin-spin interaction is not observed. ATP spin-labeled at C-8 is bound about 10-fold tighter than the corresponding AMP derivative. Four equivalents of ATP are bound per tetramer, but it exhibits no spin-spin interactions. It is concluded that the structure of the pyridine moiety of the coenzymes plays a role in orienting the adenine ring and, thus, affects the cooperativity. The N6 derivative of NAD+ also shows spin-spin interaction; however, only data for the C-8 derivatives are shown in detail. PMID- 6289907 TI - Parathyroid hormone stimulation of renal phosphoinositide metabolism is a cyclic nucleotide-independent effect. AB - The effects of parathyroid hormone (PTH) and cyclic nucleotides on renal phosphoinositide metabolism were studied using cortical tubules isolated from dog kidneys. PTH stimulated the initial rates of 32Pi incorporation into phosphatidylinositol 4'5'-diphosphate, phosphatidylinositol 4'-monophosphate, phosphatidylinositol and phosphatidic acid. PTH also caused a 45-55% increase in the actual tissue levels of these phospholipids by 5 min of incubation. By 30 min of incubation, the levels of 32Pi incorporated were similar in PTH and control flasks, but the actual levels of the phosphoinositides remained elevated, indicating stimulation of their turnover. Additional evidence of increased turnover of phosphatidylinositol was obtained from tubules pre-incubated with myo [2-3H]inositol. PTH stimulated a rapid short-lived decrement in [3H]phosphatidylinositol while total phosphatidylinositol levels increased, indicating increased turnover rates of phosphatidylinositol. In tubules pre incubated with [14C]arachidonic acid, indicating utilization of diacylglycerol produced during turnover for resynthesis of phosphatidic acid and phosphoinositides. Cyclic nucleotides and phosphodiesterase inhibition failed to reproduce the effect of PTH on phosphoinositide metabolism. These studies indicate that PTH stimulates renal phosphoinositide metabolism through a mechanism independent of cAMP which results in a net synthesis of the phosphoinositides. PMID- 6289908 TI - The role of lecithin:cholesterol acyltransferase for the low density lipoprotein composition and specific binding to the B receptor. PMID- 6289909 TI - Separate monoacylglycerol and diacylglycerol acyltransferases function in intestinal triacylglycerol synthesis. PMID- 6289910 TI - A saturable, high-affinity binding site for human low density lipoprotein on freshly isolated rat hepatocytes. AB - Freshly isolated rat hepatocytes bind the solely apolipoprotein B-containing human low density lipoprotein (LDL) with a high-affinity component. After 1 h of incubation less than 30% of the cell-associated human LDL is internalized and no evidence for any subsequent high-affinity degradation was obtained. Scatchard analysis of the binding data for human 125I-labeled LDL indicates that the high affinity receptor for human LDL on rat hepatocytes possesses a Kd of 2.6 x 10( 8)M, while the binding is dependent on the extracellular Ca2+ concentration. Competition experiments indicate that both the apolipoprotein B-containing lipoproteins (human LDL and rat LDL) as well as the apolipoprotein E-containing lipoproteins (human HDL and rat HDL) do compete for the same surface receptor. It is concluded that hepatocytes freshly isolated from untreated rats do contain, in addition to the earlier described rat lipoprotein receptor which does not interact with human apolipoprotein B-containing LDL, a high-affinity receptor which interacts both with solely apolipoprotein B-containing human LDL and apolipoprotein E-containing lipoproteins. PMID- 6289911 TI - Incorporation of exogenous fatty acids into phospholipids by cultured hamster fibroblasts. Effect of SV40 transformation. AB - In situ incorporation of two saturated (palmitic, 16:0; stearic, 18:0) and three unsaturated fatty acids (oleic, 18:1; linoleic, 18:2; arachidonic, 20:4) into the four major phospholipids, sphingomyelin, PC, PI and PE, was followed. Transformed cells incorporated unsaturated fatty acids more rapidly, whereas no significant differences were found concerning saturated fatty acids. In vitro determination of phospholipid acylation showed that incorporation of coenzyme A-activated forms of two saturated fatty acids (16:0 and 18:0) and one unsaturated fatty acid (18:1) into phospholipids was increased in transformed cells. Comparison of results obtained in situ and in vitro strongly suggests that incorporation of fatty acids into phospholipids in cultured cells is not limited by acyltransferase activities. PMID- 6289912 TI - Preparative isolation of polyphosphoinositide fractions from ox brain. AB - A simple preparative method for chromatographic isolation of pure fractions of di and triphosphoinositides (1-phosphatidylinositol 4-phosphate and 1 phosphatidylinositol 4,5-bisphosphate) from ox brain is described. Polyphosphoinositide fractions have been obtained by ion-exchange chromatography of the lipid extract using gradient elution with 0-0.6 M ammonium acetate in chloroform/methanol/water (20:9:1) from a DEAE-cellulose column. Before chromatography, divalent metal ions were removed from the lipid extract by passing through a Dowex-50 (H+) column and lipids were converted to the sodium salt by neutralisation with sodium hydroxide in methanol solution. After chromatography, fractions of di- and triphosphoinositides were precipitated in methanol/water mixture (1:1) by evaporation in a vacuum to a final concentration of about 4 M ammonium acetate. Necessary salts of di- and triphosphoinositides were obtained by passing the ammonium salts of the lipids through Dowex-50 (H+) and neutralising with corresponding base in methanol solution. About 0.35 mmol of diphosphoinositide and 0.63 mmol of triphosphoinositide were obtained from 1 kg of wet ox brain tissue. PMID- 6289913 TI - Turnover, change of composition with rate of cell growth and effect of phenylxyloside on synthesis and structure of cell surface sulfated glycosaminoglycans of normal and transformed cells. AB - The synthesis of sulfated glycosaminoglycans was analysed in mouse fibroblasts during the transition from exponential growth to quiescent monolayers. 'Normal' Swiss 3T3 fibroblasts were compared with SV40 transformed 3T3, C6, ST1 and HeLa cells. p-Nitrophenyl-beta-D-xyloside, an artificial acceptor for glycosaminoglycans synthesis, was used as a probe. Exponentially growing 'normal' 3T3 cells synthesized both dermatan sulfate and chondroitin 4-sulfate, retaining the latter and releasing the former to the medium. Upon reaching quiescence these cells switched to retention of dermatan sulfate and release of chondroitin 4 sulfate. SV3T3 cells synthesized several fold less sulfated glycosaminoglycans than 'normal' 3T3. Even though SV3T3 cells are able to synthesize dermatan sulfate, they only retained chondroitin 4-sulfate, never switching to retention of dermatan sulfate. These results indicated that the transition from rapidly proliferating to resting G0 state in normal cells is accompanied by a switch from chondroitin-sulfate rich to dermatan-sulfate-rich cells. This switching was not observed with transformed cells, which are unable to enter the G0 state. Phenylxyloside caused a several fold increase in glycosaminoglycans released to the medium in both cell types, but it did not interfere with either growth rate or cell morphology. Particularly the phenylxyloside treatment led to an increase of more than 10-fold in production of dermatan and chondroitin sulfate by SV3T3, C6, ST1 and HeLa cells. This demonstrated that transformed cells have a high capacity for glycosaminoglycan synthesis. Analysis of enzymatic degradation products of glycosaminoglycans, synthesized in the presence of phenylxyloside, by normal and transformed cells, led to the finding of 4- and 6-sulfated iduronic and glucuronic acid-containing disaccharides. This result indicated that the xyloside causes the synthesis of a peculiar chondroitin sulfate/dermatan sulfate, in both normal and transformed cells. PMID- 6289914 TI - Negative control of norepinephrine on the thyroid cyclic AMP system. AB - Negative control on the thyroid cyclic AMP system has been studied. The increase of cyclic AMP levels induced by TSH in dog thyroid slices and its consequent secretion were inhibited by norepinephrine. This effect was different from the previously described activation of cyclic AMP disposal by acetylcholine: it was not calcium-dependent, was observed in the presence of isobutyl methylxanthine and was not inhibited by atropine. The inhibitory action of norepinephrine was abolished by phentolamine but not by L-propranolol. Clonidine and epinephrine also markedly inhibited the elevation of cyclic AMP levels, but phenylephrine did not. The inhibitory effect of norepinephrine and clonidine was abolished by yohimbine but not by prazosin. These results suggest that the effect of adrenergic agents on dog thyroid follicular cells is mediated by alpha 2 receptors. Similar results were obtained on dog thyroid adenylate cyclase activity: norepinephrine diminished the activation of adenylate cyclase induced by TSH, in a sodium-dependent process. This inhibition was abolished by phentolamine and yohimbine, but not by L-propranolol and and prazosin. This shows that the negative alpha 2-adrenergic effect bears directly on adenylate cyclase. PMID- 6289915 TI - The roles of degranulation and superoxide anion generation in neutrophil aggregation. AB - Human neutrophils when exposed to appropriate stimuli aggregate, generate O(2) and secrete lysosomal constituents. To determine whether a causal relationship may exist between these responses neutrophils were exposed to either N-formyl methionyl-leucyl-phenylalanine, phorbol myristate acetate, or the two calcium ionophores, A23187 and prostaglandin Bx. Each agent elicited all of the above responses. The concentrations required to elicit the aggregation of 30 . 10(6) neutrophils/ml were comparable to that required for O(2) generation or lysozyme release. In a series of experiments designed to dissociate these responses, cells were suspended in a concentration too dilute (3 . 10(6) neutrophils/ml) to permit aggregation to occur. O(2) generation and lysozyme release was measurable and varied in a dose-dependent fashion to the concentration of stimulus. In a second series of experiments, neutrophils were treated with 4,4'-diisothiocyanostilbene 2,2'-disulfonic acid to inhibit degranulation without affecting O(2) generation. Aggregation was inhibited in a parallel fashion with lysozyme release. When detectable O(2) was removed from the medium by superoxide dismutase and catalase, aggregation and lysozyme release unaffected showing that aggregation can not be due to the presence of O(2) or its products in the extracellular medium. Neither aggregation of resting cells nor augmentation of fMet-Leu-Phe-induced aggregation was observed when cells were exposed to either supernatants of degranulated neutrophils or constituents of specific granules (lysozyme, lactoferrin). Kinetic analysis showed that in the absence of cytochalasin B degranulation preceded aggregation, while in its presence aggregation preceded degranulation. PMID- 6289917 TI - [Reliability of enzyme systems and molecular mechanisms of aging]. AB - A stochastic model of animal mortality and ageing is proposed. In a living being there is a "biological clock", constructed from Q kinds of functional elements (genes), The reliability of functioning of these genes determines the biological age of the being. Oxidative processes in the cells are accompanied by random failures of electron transfer enzyme systems, which result in appearance of free superoxide radicals. Failures of electron transfer enzymes and of defence enzymes lead to accumulation of functional damages in the genes of the "clock". In terms of the model experimental data of gerontology (Gompertz's law, Strehler-Mildvan's compensation effect, etc) are easily explained. The value of Q for the man is estimated to be of the order of 10. Some possible mechanisms of action of "geroprotectors" have been discussed. PMID- 6289916 TI - [D2O as a modifier of ionic specificity of Na, K-ATPase]. AB - Effect of heavy water D2O on the rate of hydrolysis of ATP and pNPP by Na,K ATPase was studied. Heavy water of high concentration inhibits the rate of ATPase reaction in all the studied ratios of the ions Na/K at constant ionic strength 150 mM. Activation of the enzyme was observed in the solution with low concentration of heavy water (less than 5%). The value of isotope effects depended on the ratio between sodium and potassium ion concentrations in the medium. At low temperature no activation of the enzyme with heavy water in low concentration was observed. Substitution of usual water for the heavy one was accompanied by a decrease of apparent constants of enzyme activation with sodium and potassium ions. During pNPP hydrolysis with Na,K-ATPase an increase of reaction rate in the medium with heavy water was observed. Substitution of potassium ions by cesium resulted in an increase of isotope effects during ATP and pNPP hydrolysis. Analysis of isotope effects in terms of the molecular model of sodium pump proposed permits a conclusion that the isotope effects of heavy water are explained by its influence as a solvent, the binding centres of potassium and sodium ions are localized in different regions of the enzyme differing in physico-chemical properties. The structure of sodium centres is controlled by hydrogen bonds, and of potassium ones--by hydrophobic interactions; the transport of ions by the enzyme is accompanied by dehydration of ions. PMID- 6289918 TI - [Study of organo-mineral gels using viscosimetry and electron spin exchange]. AB - Water systems with different content of soil organo-mineral complex (OMC) were studied by viscosimetry in a capillary and by the spin probe. The viscosimetric data suggest the existence of structural reconstructions in the range of 32 degrees C. The ESR data are analysed assuming simultaneous existence of two mechanisms of relaxation of spin exchange and dipole-dipole interaction. Localization of the hydrophile probe (tanole) in the aqueous region of the spectra was stated. The facts that the rotationary diffusion of the probe is independent of viscosity and that translation diffusion of the probe strongly depends on viscosity are in agreement with the assumption concerning the formation of trimeric net of couplings in the systems with OMC content above 4%. PMID- 6289919 TI - [Ion channels, induced by amphotericin B introduced unilaterally into lipid bilayers]. AB - Single ionic channels of approximately 10 pS in magnitude and approximately 100 ms duration (in 2 M KCl solution) were recorded when amphotericin B (AB) was added to one side of a lipid bilayer. Using blocking ions it has been shown that these channels are asymmetrical half-pores (similar to those postulated by Marty and Finkelstein) which are capable of forming long-living symmetrical pores if AB is added to both sides of the membrane. PMID- 6289920 TI - [Age changes of structural state of synaptic membranes of brain]. AB - In the brain synaptic membranes from old rats (24-26 months) in comparison with the mature ones (6-7 months) an increase was shown in disintegration rate with DDS Na, mobility of 5-, 16-doxyl-stearate spin probes and in thermosensitivity of acetylcholinesterase. The total number of SH-groups decreased. While protein composition remained constant lysophosphatidylcholine and saturated fatty acid content in old animals increased. The age-dependent structural rearrangement of synaptic membranes is suggested. PMID- 6289921 TI - [Formation of free radicals at photoreduction of allophycocyanin B]. AB - A photoinduced ESR signal is detected in the presence of 10(-2) M of dithionine in phycobilisomes and in fractions enriched in allophycocyanin B. The signal is characterized by the line width of 12G and g-factor of 2.004. It is demonstrated that the appearance of the free radicals is related to photoreduction of allophycocyanin B. PMID- 6289922 TI - [2 components of calcium current in mollusk neurons]. PMID- 6289923 TI - Regulation of prostaglandin E2 and F alpha productions by rat brown fat during the perinatal period; effects of ambient temperature. PMID- 6289925 TI - [Comparative enzymatic study on the structure of chromatin in the nuclei of the growing microplasmodium of Physarum polycephalum]. AB - The structure of chromatin in the growing myxomycetes P. polycephalum can be interpreted on the basis of three types of substructure: (i) soluble in a low ionic strength buffer and 2 mM EDTA (I), (ii) soluble in 0.6 M NaCl (II) and (iii) non-soluble and nuclease-insensitive (III). The ratio of these structures in the nuclear pellet obtained after autolysis was found to be 71 : 0, 4 : 12 for the young and 7.7 : 62 : 20.6 for the old cultures, respectively. The ageing of the culture involves transition of substructure I to substructure II. The substructures differ in the nuclease sensitivity. The specific features of enzyme fragmentation of chromatin from P. polycephalum microplasmodium nuclei are discussed. PMID- 6289924 TI - [Peculiarities of specific binding of serotonin by blood leukocytes, peritoneal cells and synaptosomes of mice]. AB - The properties of serotonin-active sites were studied on peritoneal cells, blood leukocytes and synaptosomes of mice (CBA line). Treatment of cell suspensions with EDTA, ouabain, strophanthin, 2,4-dinitrophenol, dithiothreitol and trypsin demonstrated that serotonin binding by peritoneal cells and leukocytes depends on bivalent ions and K+, Na+-pump operation, requires energy and intact disulfide bonds and is determined by a protein structure. ATP and ADP were found to inhibit amine adsorption by peritoneal cells. These cells specifically bind ATP 10 times more intensively than leukocytes. The data obtained are suggestive of differences in the composition of serotonin-active structures of blood leukocytes, peritoneal cells and synaptosomes. PMID- 6289926 TI - Pituitary and ovarian luteinizing hormone releasing hormone receptors during the estrous cycle, pregnancy and lactation in the rat. AB - Female Sprague-Dawley rats were decapitated at various stages of the estrous cycle, pregnancy, lactation and following ovariectomy. Anterior pituitary and ovarian tissues were collected and assayed to quantify luteinizing hormone releasing hormone (LHRH) receptors. No changes were noted in receptor affinity either between tissues or physiological stages studied. Pituitary LHRH receptor concentrations and content were greater (P less than 0.05) during diestrus II and proestrus than during estrus. Pituitary LHRH receptor concentrations and content during pregnancy were not different from those during estrus, however, a significant decrease was noted in pituitary LHRH receptor content and concentrations during lactation compared to estrus. Ovarian LHRH receptor content did not change with stage of reproduction (P less than 0.05). There was, however, a decrease (P less than 0.05) in ovarian LHRH receptor concentrations at Week 3 of pregnancy and Week 1 of lactation which was possibly due to the increase ovarian weight noted at both these physiological stages. There was no correlation (P less than 0.1) between ovarian and pituitary LHRH receptor numbers (r = 0.096). These findings suggest that the internal mechanisms which control changes in pituitary LHRH receptor numbers do not control ovarian LHRH receptor numbers. PMID- 6289927 TI - Differential action of sulfated glycosaminoglycans on follicle-stimulating hormone-induced functions of cumuli oophori isolated from mice. PMID- 6289928 TI - Model for the adsorption of cytochrome c at the water-mercury interface. PMID- 6289930 TI - [Effect of thyroidectomy and hyperthyroidism on Ca2+/2H+-antiporter activity in rat liver mitochondria]. AB - The manifestation of Ca2+/2H+ antiporter activity in rat liver mitochondria was shown to be inhibited in thyroidectomy and stimulated in hyperthyroidosis. Experiments with measuring the kinetics of the swelling of deenergized mitochondria in isoosmotic solutions Ca (NO3)2, pH 8.1 demonstrated inhibition of the swelling of liver mitochondria during thyroidectomy and stimulation because of administering thyroid hormones in vivo. During thyroidectomy, the phosphate induced swelling of rat liver mitochondria was powerfully inhibited. Meanwhile administration of thyroxine to rats stimulated the swelling of mitochondria. PMID- 6289929 TI - Spin label study of erythrocyte membrane submitted to a bending stress. AB - Red blood cells were submitted to a bending stress by incubation in hyperosmolar media and their membrane molecular structure studied by a fatty acid paramagnetic label which probes its hydrophobic part. Measurement of the apparent rotation frequency of the label, and numerical analysis of the spectrum shapes indicate that a high fluidity phase appears in the stressed membrane. The physiological implications of this finding are discussed in the light of the rheological properties of the circulating red blood cell. PMID- 6289931 TI - [Influence of lithium hydroxybutyrate on the electroencephalographic effects of fenamin]. AB - Lithium hydroxybutyrate (10 mg/kg) prevents the amphetamine-induced EEG arousal and amplitude frequency alterations in the motor and visual cortex, posterior hypothalamus, midbrain reticular formation, and caudate nucleus but potentiates the action of the psychostimulant on the EEG of the hippocamp and amygdala. The response to the light flickering rhythm in the visual cortex remains within initial upon concurrent administration of both the drugs. PMID- 6289932 TI - [Ultrastructural characteristics of the parenchymatous cells in breast cancer]. AB - Electron microscopy was used to examine ultrastructure of parenchymatous cells of lobular mammary carcinoma and preservation of an ability of these cells to organ specificity. Ultrastructural characteristics of the cells of intact mammary gland lobules and parenchymatous cells of infiltrative carcinoma were studied and compared. It was shown that lobular mammary carcinoma has dissimilar cellular composition. It is represented by the cells with ultrastructural features characteristic for the three cell types occurring in the intact mammary gland (secretory epithelium, myoepithelium and cambial cells). As regards ultrastructure, parenchymatous cells of lobular mammary carcinoma, like the cells of the normal lobule, are at varying levels of differentiation and in diverse phases of the functional status. PMID- 6289933 TI - [Fibronectin and fibronectin receptors on the surfaces of polymorphonuclear leukocytes]. AB - Interaction between human neutrophils and fibronectin bound to gelatin-Sepharose granules was studied. It was found that fibronectin following cell-induced dissociation from gelatin-Sepharose caused neutrophil aggregation which was inhibited by EDTA or pretrypsinization of the cells. Besides, polymorphonuclear lymphocytes were found to be capable to adhere to and to spread on gelatin Sepharose granules. This process was enhanced in the presence of heparin and inhibited by neutrophil pretrypsinization. The conclusion has been drawn on simultaneous existence of fibronectin and fibronectin receptors on the surface of neutrophils. A hypothesis is advanced of the existence in the fibronectin molecule of two independent binding sites, one being responsible for cell attachment and the other for gelatin binding. PMID- 6289934 TI - [Functional maturation of the human corticotropin releasing factor adrenocorticotropic hormone system during intrauterine life. An in vitro study]. AB - Radioimmunoassay was used to determine ACTH secretion by cultured hypophyses of human fetuses from the 6th to the 30th week of intrauterine life and their responsiveness to hypothalamic extracts obtained from adult animals. CRF-like activity in the human hypothalamus was measured within the 6th to the 32nd week of prenatal development from changes in ACTH release by cultured cells of the adult rat hypophysis. It was established that starting from weeks 6-7 of embryogenesis, the human fetal hypophysis is capable of synthesizing and secreting immuno-reactive ACTH in vitro. The human fetus hypothalamus of the first trimester of gestation contained no CRF-like substance. The fetus hypothalamus of the second and third trimesters of pregnancy manifested a considerable amount of CRF-like substance. It is suggested that CRF appears at the end of the first trimester of pregnancy. PMID- 6289935 TI - [Chronobiological studies of the antimutagenic action of methyluracil in cytogenetic lesions induced in mice by the vaccinal strain of the poliomyelitis virus]. PMID- 6289936 TI - [Role of cAMP in regulating hippocampal neuronal reactivity in vitro]. AB - The author studied the effect of the elevated cAMP content on the efficacy of the synaptic systems of the hippocamp. The population spike (PS) response to Shaffer collateral electric stimulation was recorded in the CA1 field. The PS amplitude served as criterion of cell reactivity. Use was made of dibutyryl-cAMP (db-cAMP), an analog of cAMP, well penetrating the membrane, and of 8-/Cl acetylaminoethylthio/-cAMP, an inhibitor of phosphodiesterase (PDE) of irreversible action, leading to cAMP accumulation by the cell. Introduction of db cAMP into the bath medium evoked an abrupt increase in the PS amplitude, followed by gradual diminution of the response until complete depression PDE inhibitor evoked a gradual and irreversible increase of the PS amplitude. The data suggest that the secondary messenger cAMP plays an important role in synaptic processes occurring in the hippocamp. PMID- 6289938 TI - [Participation of cGMP in realizing the effect of cholecystokinin-pancreozymin on the bile-excreting function of the human liver]. AB - The paper concerns studying the participation of cyclic nucleotides in the mechanisms of action of cholecystokinin on gallbladder function in man. Cyclic nucleosides (cAMP and cGMP) were identified by radioimmunoassay in the duodenal contents obtained from men in response to intravenous injection of cholecystokinin. The data obtained suggest that the action of cholecystokinin on gallbladder function in man is mediated via cGMP, whereas cAMP is not implicated in the effect of of cholecystokinin on gallbladder function in man. Disturbances in cyclic nucleotide systems may be viewed in the light of the evidence obtained as a possible pathogenetic factor in the development of gallbladder dyskinesia. PMID- 6289937 TI - [Lipid peroxidation and the myeloperoxidase activity of neutrophilic leukocytes in shereshevskyi-Turner syndrome]. AB - The level of lipid peroxides, activity and thermostability of myeloperoxidase have been studied in neutrophil extracts from the peripheral blood of patients with the Shereshevsky-Turner syndrome and in normal subjects. A statistically significant rise in the level of lipid peroxides and respective decrease in the activity and thermostability of myeloperoxidase have been revealed in the patients as compared with the control group. These findings point to functional inferiority of neutrophils. It is probable that at the basis of the disorders seen there lie an abnormal set of sex chromosomes and upset hormonal regulation determined by the abnormality. PMID- 6289939 TI - [Genetic characteristics of plasmid pAP53 derepressed in transfer functions detected in the cells of an opportunistic E. coli strain]. AB - A study was made of plasmid pAP53 derepressed as regards transfer functions (Tra functions) detected in E. coli strain cells, serogroup 0128, after its labeling with transpozones Tn1 and Tn9. The compatibility tests demonstrated that the plasmid belongs to the incompatibility group FIII and is partially incompatible with the group FII reference-plasmid. Plasmid pAP53 is unable to inhibit Tra functions of plasmid F'lac and is not inhibited by the fin type genetic regulation on the OP group plasmids under study. At the same time Tra-functions of plasmid pAP53 are inhibited in the presence of pAP41 plasmid, which indicates that this plasmid has a special type of genetic regulation. PMID- 6289940 TI - [Crystalline lens "tumor immunity"]. AB - Three series of experiments on 72 rabbits have shown that lens tumor immunity is not related to the anaerobic nature of its energy metabolism and is not a consequence of lens capsule impermeability for chemical cancer genes. Ionizing radiation causes cataracts without malignancy of lens epithelial cells. PMID- 6289941 TI - Biochemical correlates of the differential sensitivity of subtypes of human leukemia to deoxyadenosine and deoxycoformycin. AB - Leukemic cells incubated in vitro with 2'-deoxyadenosine (dAdo) plus an inhibitor of adenosine deaminase, 2'-deoxy-coformycin (DCF), show different metabolic responses depending on the histologic and immunologic type of the leukemia. Leukemic cells were obtained from 54 patients with acute lymphoblastic leukemia (ALL), 9 with myeloid or nonlymphoblastic leukemia, 3 with chronic lymphocytic leukemia (CLL), and 3 with lymphoma. There was a wide variation in the LD50, the concentration of dAdo that caused 50% inhibition of the incorporation of 3H thymidine into cells in the presence of 20 microM DCF. T-cell leukemia specimens were much more sensitive to dAdo than were specimens of pre-B-ALL and null-ALL. In leukemic cells that had been incubated with 14C-dAdo plus DCF, a good correlation was observed between the LD50 and the ratio of 14C-deoxyATP to ATP (correlation coefficient for the fit to a hyperbola = 0.853). The accumulation of deoxyATP by the leukemic cell specimens was correlated best with the activity of ecto-ATPase, less well with cytoplasmic 5'-nucleotidase and deoxyadenosine kinase, and poorly with adenosine deaminase and ecto-5'-nucleotidase. The clinical response to DCF therapy of a patient with T-ALL and another with pre-B ALL was consistent with the in vitro metabolic response of their cells to DCF and dAdo. PMID- 6289942 TI - The inhibitory effects of exogenous arachidonic acid on rabbit platelet aggregation and the release reaction. AB - Although arachidonic acid causes rabbit platelet aggregation and the release of granule contents in suspensions of washed platelets when used in concentrations of approximately 50-300 microM, higher concentrations (500 microM) cause neither aggregation nor release. Suspensions of platelets from rabbits wee exposed to arachidonic acid (250 microM) for 15 min, allowed to recover in the presence of PGE1 for 30 min, washed, and resuspended; in some experiments, the platelets were treated with aspirin before being exposed to arachidonic acid. Aggregation of platelets pretreated with arachidonic acid was inhibited in response to ADP; this effect was greater with the non-aspirin-treated platelets and persisted for at least 4 hr after resuspension. The association of 125I-fibrinogen with the platelets as a result of ADP stimulation was also inhibited. Aggregation and release of granule contents in response to collagen and low concentrations of thrombin was inhibited, but the inhibition could be overcome by higher concentrations. Thrombin induced further release of granule contents from platelets exposed to arachidonic acid without pretreatment with aspirin. Platelets that had been exposed to arachidonic acid, either with or without pretreatment with aspirin, did not aggregate or undergo further release upon stimulation with arachidonic acid after they were washed and resuspended. Inhibition of the lipoxygenase pathway with eicosatetraynoic acid (ETYA) or nordihydroguaiaretic acid (NDGA) did not affect the inhibition caused by arachidonic acid, so it is unlikely that a product of this pathway is responsible for the inhibition. Mixing experiments indicated that the pretreated platelets did not form a thromboxane-A2-like activity, and that they were unresponsive to aggregation and release induced by products formed from arachidonic acid. Experiments with 3H-arachidonic acid showed that after 45 min of incubation with platelets, only 1.1% of the 3H-arachidonic acid remained as free arachidonic acid in the platelets. Although cyclic-AMP was slightly increased 1 min after the addition of arachidonic acid, the cyclic-AMP concentration was the same as that of control platelets after the platelets were washed and resuspended, indicating that increased cyclic-AMP is not likely to be responsible for the persistent inhibitory effect. Thus, the inhibitory effect of pretreatment with arachidonic acid is a general effect on responses to a variety of aggregating agents that act through different mechanisms, and the inhibition is not related to thromboxane-A2 formation. The possibility of membrane perturbation resulting in the unavailability of receptors may explain the persistent inhibitory effect, but the responsible reactions have not been identified. PMID- 6289944 TI - Hemolytic anemia in hereditary pyrimidine 5'-nucleotidase deficiency: nucleotide inhibition of G6PD and the pentose phosphate shunt. AB - We evaluated the erythrocytes of two patients with hereditary pyrimidine 5' nucleotidase deficiency. Significant findings included an increased reduced glutathione content, increased incubated Heinz body formation, a positive ascorbate cyanide test, and decreased intraerythrocytic pH. The pentose phosphate shunt activity of the patients' red cells as measured by the release of 14CO2 from 14C-1-glucose was decreased compared to high reticulocyte controls. Glucose 6-phosphate dehydrogenase (G6PD) activity in hemolysates from control erythrocytes was inhibited 43% by 5.5 mM cytidine 5'-triphosphate (CTP) and 50% by 5.5 mM in uridine 5'-triphosphate (UTP) at pH 7.1. CTP was a competitive inhibitor for G6P (Ki = 1.7 mM) and a noncompetitive inhibitor for NADP+ (Ki = 7.8 mM). Glutathione peroxidase, glutathione reductase, and 6-phosphogluconate dehydrogenase were not affected by these compounds. Pentose phosphate shunt activity in control red cell hemolysate at pH 7.1 was inhibited to a similar degree by 5.5 mM CTP or UTP. Since the intracellular concentrations of G6P and NADP+ are below their KmS for G6PD, these data suggest that high concentrations of pyrimidine 5'-nucleotides depress pentose phosphate shunt activity in pyrimidin 5'-nucleotidase deficiency. Thus, this impairment of the pentose phosphate pathway appears to contribute to the pathogenesis of hemolysis in pyrimidine 5'-nucleotidase deficiency hemolytic anemia. PMID- 6289943 TI - Con-A-stimulated superoxide production by granulocytes: reversible activation of NADPH oxidase. AB - Stimulation of granulocyte (PMN) superoxide (O2-) production by concanavalin-A (Con-A) can be monitored continuously in the spectrophotometer. Both the rate of activation and final activity of the O2--generating system is dependent on the concentration of Con-A. Alpha methylmannoside (alpha MM) can prevent Con-A, but not phorbol myristate acetate (PMA) or zymosan, induced O2- production. Alpha MM inhibits both the rate of activation and the final rate of O2- production. When alpha MM is added after the attainment of a maximal rate of O2- production with Con-A, O2- production continues for another minute before it ceases. When PMA is added to such treated cells, it restores O2- production. Although the inhibition of O2- production by alpha MM on previously activated cells requires time, most of the bound concanavalin-A is removed immediately after the addition of alpha MM. Treatment of cells with L-1-tosylamido-2-phenylethyl-chloromethyl ketone (TPCK) prevents activation of PMN by Con-A to a greater extent than it does for either PMA or zymosan. TPCK has no effect on the binding of Con-A. TPCK, when added after Con-A, will inactivate O2- production by the cells. The addition of PMA after TPCK treatment restores O2--generating activity. Membrane-enriched particles from PMN activated with Con-A, alpha MM, and PMA demonstrate that the change in O2- production seen by whole cells is due to an alteration of the activity of the NADPH oxidase. Thus, Con-A stimulation of human PMN O2- production can be prevented and reversed by the addition of either alpha MM or TPCK and that PMA can reactivate Con-A and either alpha MM- or TPCK-treated cells. The activation, inactivation, and reactivation occur as a result of changes in the plasma membrane NADPH-dependent O2--generating enzyme. PMID- 6289945 TI - Abnormal biochemistry of subcellular membranes isolated from nonvascular smooth muscles of spontaneously hypertensive rats. AB - Microsomal fractions enriched in plasma membranes and endoplasmic reticulum were isolated from stomach fundus and vas deferens from age-matched Okamoto spontaneously hypertensive (SHR) rats and corresponding Kyoto-Wistar normotensive rats (KWR). Alterations of several enzyme activities and Ca2+ accumulation of the isolated microsomal fraction from these nonvascular smooth muscles provide direct evidence of abnormal smooth muscle membrane biochemistry in SHR. Decreased Ca2+ accumulation in the presence of but not in the absence of adenosine triphosphate by the microsomal fractions of both fundus and vas deferens from SHR is consistent with previous findings using plasma membranes from vascular smooth muscles from SHR and cannot be explained in terms of adaptation induced by elevation of blood pressure in SHR. Defective Ca2+ handling now observed in both vascular and nonvascular smooth muscles from hypertensive animals not only provides a cellular basis for the altered reactivity and contractility of smooth muscles observed in SHR, but also supports the hypothesis that spontaneous hypertension is associated with a generalized widespread alteration in smooth muscle membrane fraction. PMID- 6289946 TI - [Influence of B locus on avian viral induced tumours]. AB - Since early in the century Avian Cancers were described as induced by viruses which later were known of DNA and RNA types. The susceptibility of birds was found different according to the genetic lines of the chickens and specially to the B locus blood group. Since the B locus of birds was strongly associated with the Major Histocompatibility Complex (MHC) it was of interest to review the last reports on the influence of the B locus on viral induced tumours. In Marek's Disease due to a DNA virus (Herpes type) the B21 allele expresses the greater resistance compared with other B alleles although non-B factors could be involved as demonstrated with the lymphocyte factor Ly-4. The possible mechanisms of the influence of B locus on the resistance against Marek's Disease are discussed. The tumours induced by RNA viruses (Avian Leucosis Sarcomas) develop or regress following genetic characters closely related to MHC. Differences of resistance exist between B alleles. Complementary genes should be present to fully express the resistance. The relationship between MHC-B locus and resistance to tumors stimulates the actual assays. Since a number of parameters remain still unknown further researches should be done in order to evidence the involved mechanisms of the resistance. PMID- 6289947 TI - Common epithelial tumors of borderline malignancy (carcinomas of low malignant potential). AB - The creation of the category of ovarian borderline tumors has been a step forward in classification because it has segregated from the general group of common epithelial cancers a subgroup with an unusually good prognosis. Of all borderline tumors, the serous variety is the least difficult for the pathologist to diagnose. The mucinous borderline tumor is not as easy to recognize when an absence of invasion is used as the sole diagnostic criterion, but evaluation of other architectural as well as cytological features is helpful in making the diagnosis. Although the treatment of the majority of serous and mucinous borderline tumors is well standardized, therapy of residual serous borderline tumor is highly controversial and the treatment for pseudomyxoma peritonei, unsatisfactory. Borderline tumors exist in the endometrioid, clear cell, Brenner, and mixed common epithelial categories, but these tumors have been too rare to date in order to clearly characterize their clinical and pathological features. It is hoped that more can be learned about this general group of tumors by a more widespread recognition of their distinctive clinico-pathologic features and by their acceptance as a special subgroup of ovarian cancer. Their behavior differs strikingly from that of other types of malignant ovarian tumors and it is clear that their management should differ accordingly. PMID- 6289948 TI - [Ovarian cancer: therapeutic indications in the adult]. PMID- 6289949 TI - Enzymatic hydrolysis of concentrated diazinon in soil. PMID- 6289950 TI - Field persistence studies with triallate and trifluralin both singly and in combination with chloramben. PMID- 6289951 TI - Management of bacterial infections in haematological patients. PMID- 6289952 TI - Inhibition of ionic currents in frog node of Ranvier treated with naloxone. AB - 1 Myelinated nerve fibres of frog sciatic nerve were investigated under current and voltage clamp conditions. In the presence of 68 microM external naloxone, the action potential was completely, though progressively, blocked within 15 min of drug superfusion. The resting potential remained constant. 2 Under voltage clamp conditions both peak Na+ and steady-state K+ currents were decreased reversibly by external naloxone. Both currents were reduced in a dose-dependent manner but, whereas sodium current was affected by the smallest concentrations of naloxone (1.3 up to 12.5 microM), potassium current was decreased only by higher concentrations (25 up to 112 microM). 3 The time-course of development of the effect on both Na+ and K+ currents after exposure to 112 microM naloxone (a concentration giving more than 50% of decrease) showed that the effect develops quickly within the first 2 min of exposure to the drug, but afterwards both currents continue to fall more slowly, though progressively. 4 Experiments with constant test pulses to Em = - 10 mV and conditioning prepulses of various amplitudes, showed that the Na inactivation curve, h infinity (Em), was shifted in a negative direction along the potential axis; the shape of the curve was also slightly changed in the presence of naloxone since the shift was larger near the top of the curve. All the observed effects were reversible after returning to the standard Ringer solution. 5 Internal naloxone (less than 0.2 mM) reduced the amplitude of the action potential as well as peak Na+ and steady-state K+ currents; the sodium inactivation curve, h infinity (Em), was shifted to more negative potentials. 6 A possible anaesthetic-like activity of naloxone on the nodal membrane is discussed. PMID- 6289953 TI - Desensitization of prostacyclin receptors in a neuronal hybrid cell line. AB - 1 Prostacyclin and its stable analogue, carbacyclin, bind competitively to a single population of receptors, and activate adenylate cyclase of the NCB-20 neuronal somatic cell hybrid (Kact = 40.1 nM and 96.1 nM respectively). 2 Culture of NCB-20 cells in the presence of 1 microM carbacyclin for 4 to 16 h results in a progressive decrease in the prostacyclin-dependent activation of adenylate cyclase in cell homogenates with an increase at 16 h of the Kact from 64.1 nM to 174.0 nM and decrease in the maximum adenylate cyclase activation from 41.2 to 15.1 pmol cyclic AMP min-1 mg-1 protein. 3 The prediction that the apparent decrease in affinity in the prostacyclin-dependent activation of adenylate cyclase was secondary to a reduction in receptor numbers was tested directly by measuring binding of [3H]-prostacyclin to membranes of cells exposed to carbacyclin for 16 h. This showed an actual decrease in affinity of the prostacyclin-receptor interaction, as well as a decrease in the total receptor numbers. Thus prolonged exposure of NCB-20 cells to carbacyclin caused reductions in both receptor numbers and affinity, reflected by measurements both of binding and adenylate cyclase activation. PMID- 6289954 TI - Presynaptic gamma-aminobutyric acid receptors in the rat anococcygeus muscle and their antagonism by 5-aminovaleric acid. AB - 1 The effects of gamma-aminobutyric acid (GABA) and related drugs on the isolated anococcygeus muscle of the rat were determined. 2 GABA caused a dose-related inhibition of the electrically-evoked twitch response. 3 The maximum response to GABA was a 56.8% depression of twitch response, with an EC50 of 0.68 microM. 4 (+/-)-Baclofen mimicked the effect of GABA (EC50 0.9 microM). (+)-Baclofen was more than 100 times less active than (--)-baclofen. 5 The response to GABA was unaffected by picrotoxin or bicuculline but was antagonized by 5-aminovaleric acid (0.5) mM). 6 Our results suggest that GABAB receptors are present on motor nerve terminals in the rat anococcygeus muscle and that 5-aminovaleric acid is an antagonist of these receptors. PMID- 6289955 TI - Receptor interactions of imidazolines: alpha-adrenoceptors of rat and rabbit aortae differentiated by relative potencies, affinities and efficacies of imidazoline agonists. AB - 1 Noradrenaline and a series of imidazolines were used to characterized and differentiate the postsynaptic alpha-adrenoceptors of rat and rabbit aortae. 2 Dose-response curves in each tissue revealed marked differences in the profile of agonist activity among the compounds. Based on the ED50 values for each compound, a rank order of potency of oxymetazoline greater than noradrenaline greater than tramazoline greater than tetrahydrozoline greater than clonidine was obtained in rabbit aorta and an order of noradrenaline greater than clonidine greater than tramazoline greater than oxymetazoline was obtained in rat aorta. Tetrahydrozoline had no agonist activity in rat aorta. 3 Dissociation constants were determined for each agonist in rat and rabbit aortae. Again, differences between the tissues were observed to the extent that the rank order of affinities for the imidazolines were exactly opposite for the two tissues. In rabbit aorta the order was, oxymetazoline greater than tramazoline greater than tetrahydrozoline greater than clonidine, whereas in rat aorta it was, clonidine greater than tetrahydrozoline greater then tramazoline greater than oxymetazoline. The extremes in tissue selectivity were observed with clonidine, which had approximately 125 fold higher affinity in rat aorta, and oxymetazoline, which had approximately 4 times higher affinity in rabbit aorta. 4 The absolute values of relative efficacies of the imidazolines studied, and their rank order, also differed between the two tissues. The relative efficacies of oxymetazoline and tramazoline were more than 15 fold greater in rabbit aorta than in rat aorta. Furthermore, tetrahydrozoline had a greater relative efficacy than clonidine in rabbit aorta while the converse was true in rat aorta. 5 Differences in the rank order of potency, affinity and relative efficacy of noradrenaline and a series of imidazolines in rat and rabbit aortae indicate that the postsynaptic alpha adrenoceptors in these tissues are different. While the postsynaptic alpha adrenoceptor of rabbit aorta is clearly of the alpha 1-subtype, the exact nature of the postsynaptic alpha-receptor of rat aorta is not clear. The unique alpha receptor of rat aorta has properties of both alpha 1- and alpha 2-adrenoceptors. PMID- 6289956 TI - Radiobiological studies with therapeutic neutron beams generated by p+ leads to Be or d+ leads to Be. AB - Mammalian cells cultured in vitro were used to study the radiobiological characteristics of neutron beams generated by 43 MeV protons on beryllium or 25 MeV deutrons on beryllium. For an unfiltered beam of neutrons generated by 43 MeV p+ leads to Be the relative biological effectiveness was found to be 8-12% higher at a depth of 2 cm than at a depth of 12 cm due to the presence of a large component of low-energy neutrons. The addition of a hydrogenous filter 4 cm thick preferentially removed the low-energy neutrons from the beam and, as a result, the neutron RBE was independent of depth. There was no significant difference in the oxygen enhancement ratio between the filtered neutrons produced by 43 Mev p+ leads to Be and neutrons produced by 25 MeV d+ leads to Be; for both beams the OER value was about 1.6. PMID- 6289958 TI - Postexposure immunoprophylaxis against B virus infection. PMID- 6289957 TI - Diagnostic value of 9 am plasma adrenocorticotrophic hormone concentrations in Cushing's disease. AB - Morning plasma adrenocorticotrophic hormone (ACTH) concentrations were measured in 58 normal subjects and seven patients with pituitary-dependent Cushing's syndrome (Cushing's disease). Particular note was taken of the time of venepuncture. The range of values for the normal subjects irrespective of timing was 9-77 ng/l. The range between 9 0 am and 9 30 am was 9-24 ng/l. In the patients with Cushing's disease the ACTH concentrations were in the range 39-109 ng/l. To distinguish patients with Cushing's disease from normal subjects it is therefore important to define accurately the 9 am normal range, since these results show no overlap. PMID- 6289959 TI - Prevention of hepatocellular carcinoma by active immunisation against hepatitis B. PMID- 6289960 TI - Arterial hypertension developing 10 years after radiotherapy for Wilms's tumour. AB - Three patients developed arterial hypertension more than 10 years after receiving irradiation for Wilm's tumour. Scattered radiation appeared to have caused changes in the remaining kidney which were not severe enough to inhibit compensatory hypertrophy but which produced a rise in blood pressure at a later date. Since arterial hypertension appears to be a delayed complication of radiotherapy which is easily detected and controlled and can occur at any age long-term surveillance after successful treatment of malignant tumours in childhood is necessary. PMID- 6289961 TI - High dose of antacid (Mylanta II) reduces bioavailability of ranitidine. AB - To investigate the effect of antacid on the bioavailability and disposition of ranitidine six healthy volunteers were studied on two occasions one week apart. In the first study the received ranitidine 150 mg with 60 ml water, and in the second study they received ranitidine 150 mg plus 30 ml of an aluminium/magnesium hydroxide mixture (Mylanta II) and 30 ml water. Giving antacid reduced both the maximum plasma ranitidine concentration and the area under the curve by one third; elimination of the drug was not changed. Thus giving a high dose of antacid significantly diminished the bioavailability of ranitidine. PMID- 6289962 TI - Acyclovir. PMID- 6289963 TI - Ceftriaxone in the treatment of ordinary and penicillinase-producing strains of Neisseria gonorrhoeae. AB - Ceftriaxone, a third generation cephalosporin, was used in a single intramuscular dose with oral probenecid to treat 124 men with infections due to non penicillinase-producing Neisseria gonorrhoeae (non-PPNG) and 64 men with infections due to PPNG. Three different doses of ceftriaxone were used--125 mg, 62.5 mg, ad 32.5 mg, and 32.5. The cure rate for all PPNG infections with the different doses was 100%. The cure rate for the non-PPNG infections with ceftriaxone 125 mg was 100%; those for non-PPNG infections treated with ceftriaxone 62.5 mg and 32.5 mg were 96.2% and 97.3% respectively. The 160 strains of non-PPNG and 60 strains of PPNG isolated were all susceptible to ceftriaxone with minimum inhibitory concentrations of 0.008 microgram/ml. These results are compared with those using kanamycin 2 g. Ceftriaxone is a safe and effective treatment for PPNG and non-PPNG infections. PMID- 6289964 TI - Responses of intracellularly recorded cortical neurons to the iontophoretic application of dopamine. AB - Considering that a well-defined dopaminergic projection from the mesencephalic structures to the rat frontal cortex has been demonstrated, the purpose of this research was to study the action of iontophoretically applied dopamine (DA) on intracellularly recorded rat frontal neurons. The stimulation of the substantia nigra (SN) and the ventral tegmental area (VTA) evoked EPSP-IPSP sequences in these cells. About 50% of the tested neurons, widely distributed in all the frontal cortex, responded to DA application and no difference in the response to DA was observed between neurons with monosynaptic inputs and neurons with polysynaptic inputs. The catecholamine depolarized the cell membrane and decreased the firing rate, generally without significant changes in membrane resistance, as already observed in rat and cat striatal cells. In some neurons the decrease of the spikes preceded the membrane depolarization. Considering the complex effect of DA on the electrical properties of these neurons, these results seem to be indicative of a mechanism of action dependent on metabolic changes. PMID- 6289965 TI - Naltrexone-induced opiate receptor supersensitivity. AB - Chronic administration of the long-lived narcotic antagonist naltrexone resulted in a marked increase in brain opiate receptors. Similar changes in receptor density were observed for binding of the putative mu agonist [3H]dihydromorphine, the mu antagonist [3H]naloxone, the putative delta ligand [3H]D-Ala2,D-Leu5 enkephalin and [3H]etorphine. In addition, the sensitivity of agonist binding to guanyl nucleotide inhibition increased significantly. In contrast, no such changes in opiate binding were observed following acute administration of naltrexone. The increase in opiate receptor number following chronic naltrexone was highest in the mesolimbic and frontal cortex areas, and lowest in the dorsal hippocampus and periaqueductal gray. These results indicate a degree of plasticity in the opiate receptor system that may correlate with specific functional pathways. PMID- 6289966 TI - Angiotensin-converting enzyme in discrete areas of the rat forebrain and pituitary gland. AB - With the use of a sensitive radioisotopic method we have examined the activity of the angiotensin-converting enzyme (ACE, E.C. 3.4.15.1) in specific nuclei of the rat forebrain and in the anterior, intermediate and posterior lobes of the pituitary gland of the rat. We reported that ACE activity is heterogeneously distributed in the rat forebrain, with a 200-fold difference between the lowest and the highest values. Highest enzyme activities were found in the subfornical organ and in the posterior lobe of the pituitary gland. High ACE activity was also detected in the intermediate and anterior lobes of the pituitary gland, the caudate nucleus, and the medial habenular nucleus. Substantial activity also existed in the globus pallidus, the median eminence, the supraoptic and paraventricular nuclei, the lateral habenular nucleus and the organon vasculosum laminae terminalis. Our results demonstrate that one of the components of the renin-angiotensin system, the angiotensin-converting enzyme, is highly localized to a few discrete brain structures and the pituitary gland. These findings suggest that angiotensin II could be formed locally in some of these structures, supporting previous immunohistochemical data. PMID- 6289967 TI - Effects of sex steroids on brain beta-endorphin. PMID- 6289968 TI - Capsaicin blocks one class of K+ channels in the frog node of Ranvier. AB - Capsaicin (10-20 microM) blocks reversibly one component of the K+ current (IKf2) in frog node of Ranvier. The block is enhanced with depolarization but it is not voltage-dependent indicating that the affinity of receptor sites for capsaicin is larger with the channels in the open configuration. The results provide pharmacological evidence of the existence within the nodal membrane of two classes of fast K+ channels, in addition to the slow K+ channels. PMID- 6289969 TI - Mu and kappa opioid agonists elevate brain stimulation threshold for escape by inhibiting aversion. AB - Rats were trained to press a lever to escape electrical stimulation of the nucleus reticularis gigantocellularis and to obtain stimulation of the lateral hypothalamus. Morphine sulfate and ethylketocyclazocine (EKC) both elevated the intensity of stimulation required to sustain escape at doses which did not affect self-stimulation. Parallel dose-response lines were obtained for the two opioid agonists but the effect of EKC was more resistant to naloxone antagonism. These results suggest that both mu-and chi-sub-types of opiate receptor mediate the inhibition of supraspinally-elicited aversion. PMID- 6289970 TI - Branched projections from the nucleus prepositus hypoglossi to the oculomotor nucleus and the cerebellum. A retrograde fluorescent double-labeling study in the cat. AB - The retrograde transport of fluorescent substances was used in order to investigate divergent axon collaterals of neurons in the nucleus prepositus hypoglossi (Ph). Fast blue (FB) was injected into the flocculus, paraflocculus and/or the vermis, while nuclear yellow (NY) was injected into the oculomotor nucleus alone or combined with injections in the nucleus of Darkschewitsch, the interstitial nucleus of Cajal and the medial longitudinal fascicle. Within optimal survival time, separate populations of single-labeled neurons of both dyes were found in Ph in all cases. Double-labeled neurons were seen in the rostral Ph following FB injections into the flocculus and the paraflocculus and NY injections restricted to the oculomotor nucleus. The present findings demonstrate that many neurons in the rostral Ph give collateral branches to the cerebellum and to the oculomotor nucleus. PMID- 6289972 TI - Vasopressin-reversible increase in angiotensin-converting enzyme in specific hypothalamic nuclei of Brattleboro rats. AB - The activity of the angiotensin-converting enzyme (ACE) (kininase II, EC 3.4.15.1) was examined in 5 discrete hypothalamic nuclei of rats lacking vasopressin (homozygous Brattleboro rats, DI, di/di) and their corresponding controls (heterozygous Brattleboro rats, HZ, di/+, and Long Evans, LE, +/+ rats), with and without hormonal replacement with arginine-vasopressin (AVP). DI rats showed a vasopressin-reversible increased ACE activity when compared with LE controls, HZ rats showing intermediate activity. These changes occurred only in the supraoptic and periventricular hypothalamic nuclei, and were absent in other hypothalamic areas studied, including the paraventricular nucleus. These results provide biochemical evidence in support of previous anatomical and physiological data, for an interaction between the brain vasopressin and angiotensin systems in discrete hypothalamic nuclei, and suggest that vasopressin could regulate the formation of brain angiotensin II by modulating the activity of the converting enzyme. PMID- 6289971 TI - Age-related changes in the distribution of visual callosal neurons following monocular enucleation in the rat. PMID- 6289973 TI - Hypoglycemia-induced elevation of immunoreactive beta-endorphin level in cerebrospinal fluid in the cat. PMID- 6289974 TI - Chronic lithium administration has no effect on haloperidol-induced supersensitivity of pre- and postsynaptic dopamine receptors in rat brain. PMID- 6289975 TI - Multiple action of acetylcholine at a muscarinic receptor studied in the rat hippocampal slice. AB - Responses of hippocampal pyramidal cells to topical application of acetylcholine (ACh) were measured in the in vitro hippocampal slice preparation. ACh but not cyclic GMP produced a short-latency hyperpolarization associated with a decrease in input resistance. This was followed by a long-latency but long duration depolarization associated in some cells with an increase in input resistance. This change in resistance followed the depolarization and outlasted it by 5--20 min, until complete recovery. During the depolarization there was a reduction in magnitude of EPSPs produced by activation of the Schaffer collateral excitatory afferents. The reversal potential for the hyperpolarization was about -95 mV, and it was blocked by 4-aminopyridine. The depolarization, but not the hyperpolarization was markedly attenuated in slices maintained in low (25 degrees C) temperature. The responses to ACh were blocked by atropine but not by D tubocurarine. The hyperpolarization as well as the depolarization were present in slices treated with tetrodotoxin (TTX)., but were reduced in slices superfused with a low Ca2+-high Mg2+ medium, and in slices treated with Mn2+ and Co2+ ions. It is suggested that ACh causes a fast increase in gK+, followed by a long lasting energy-dependent depolarization associated with action potential discharges, a decrease in conductance and a suppression of EPSPs. PMID- 6289976 TI - Role of serotonin and cyclic AMP on facilitation of the fast conducting system activity in the leech Hirudo medicinalis. AB - In the nervous system of the leech Hirudo medicinalis it has been possible to study short-term plastic changes. Depression and facilitation have been demonstrated in the fast conducting system (FCS) activity; this pathway consists of a chain of electrically linked neurons present in each ganglion. In semi intact animals or in preparation of nerve cord and segments of body wall, both electrical stimulation of peripheral roots and tactile stimulation of the skin induced, after repetitive stimulation (0.1/s) a prolonged decrement of FCS response. Strong nociceptive stimulation applied onto the head or the body wall produced a sustained facilitation of the waned response. The same potentiation has been observed by perfusing the isolated ganglion with serotonin (5 x 10(-5) M). Such a potentiation is abolished by preincubation with methysergide, an antagonist of serotonin, and with imidazole, a cAMP-phosphodiesterase activator. Such an effect is mimicked by an analog of cAMP, db-cAMP. Simultaneous recordings of both T neurons (intracellularly) and FCS firing discharge showed that, during FCS response decrement, the T cell activity remained unchanged and no modification of conductance occurred, excluding therefore a detectable involvement of sensory neurons in the depression. These results suggest that short-term plastic changes of the FCS of the leech are due to a prolonged potentiation of synaptic transmission as a result of serotonin-mediated increase in cAMP. PMID- 6289977 TI - Multiple forms of phosphoprotein phosphatase and its protein inhibitors from rat brain. AB - By means of Sephadex A-50 ion exchange chromatography 5 peaks of phosphoprotein phosphatase (PPase) activity have been detected in rat brain. These molecular forms of the enzyme differ from each other by their specific activity as well as by a number of other properties--pH optima, heat and storage stability, and substrate specificity. It was shown that cyclic AMP (10(-5)-10(-6) M) decreased PPase activity of the majority of the peaks (I-III, V) and increased that of the peak IV. Ion exchange chromatography of brain tissue extracts revealed 4 peaks of PPase protein inhibitors. The results obtained on dialysis and tryptic digestion point to the protein nature of the inhibitors that differed from each other by their effect on the enzyme activity and heat stability (95 degrees C, 5 min). The significance of the specificity of separate molecular forms of PPases is discussed. PMID- 6289978 TI - Kainic acid receptor sites in the cerebellum of nervous, Purkinje cell degeneration, reeler, staggerer and weaver mice mutant strains. PMID- 6289979 TI - Trigeminal primary afferents project bilaterally to dorsal horn and ipsilaterally to cerebellum, reticular formation, and cuneate, solitary, supratrigeminal and vagal nuclei. PMID- 6289980 TI - Modulation of an inhibitory circuit by adenosine and AMP in the hippocampus. PMID- 6289981 TI - Further studies on the action of 5-hydroxytryptamine in the dorsal lateral geniculate nucleus of the cat. AB - The possibility that 5-HT has a presynaptic inhibitory influence on the optic nerve input to the LGNd has been examined. We have compared the effect of iontophoretic application of 5-HT on the excitatory responses of LGNd neurons to visual stimulation and iontophoretic pulses of ACh and DLH. In contrast to previous studies the results suggest that 5-HT has only postsynaptic depressant effects and we found no evidence for a selective inhibition of the optic nerve input. It is proposed that the difference between the present result and those previously reported may reflect the method of stimulation of the optic nerve input. The weaker excitatory responses produced with electrical stimulation and diffuse light flashes of the earlier studies may be more susceptible to the postsynaptic depressant actions of 5-HT than the powerful visually evoked responses of the present study. Our data are only consistent with a postsynaptic depressant role for 5-HT in the LGNd. PMID- 6289983 TI - The effect of elbow joint afferent discharge on transmission in forelimb flexion reflex pathways to biceps and triceps brachii in decerebrate cats. AB - The present experiments were performed to investigate the influence of elbow joint afferent discharge on the excitability of reflex arcs mediating flexion withdrawal and crossed extensor reflexes in the forelimb muscles of decerebrate cats. The excitability of flexion withdrawal and crossed extensor reflexes in forelimb muscles was shown to be modulated by elbow joint position. Flexion withdrawal reflexes were most easily elicited when the elbow was extended and crossed extensor reflexes were most easily elicited when the elbow was flexed. This modulation of transmission was not confined to reflex pathways projecting to muscles acting at the elbow but also included pathways to muscles acting at the wrist and shoulder in addition to muscles in the contralateral forelimb. The changing excitability of reflex pathways caused by elbow movement was unaltered by degloving the skin covering the joint and tenotomy of the medial head of triceps and the long head of biceps. However, modulation of reflex excitability by joint movement was totally abolished by local anaesthesia of the joint in an otherwise intact limb. Thus, the present experiments indicate that transmission in forelimb flexion reflex pathways can be powerfully influenced by elbow joint afferent discharge. PMID- 6289982 TI - Brain beta-endorphin concentrations in experimental chronic liver disease. PMID- 6289984 TI - Protein phosphorylation in isolated synaptic junctional structures: changes during development. AB - Endogenous phosphorylation in subcellular fractions enriched in synaptic plasma membranes (SPM) and purified synaptic junctions (SJ) has been examined in vitro at various stages of postnatal development. Protein kinase activity was measured using both endogenous and exogenous (histones) proteins as substrates. Protein phosphorylation that is regulated by cyclic AMP also was investigated. SPM and SJ fractions displayed large increments in kinase activities and cyclic AMP stimulated protein phosphorylation between postnatal days 5 and 15. SJ fractions exhibited a dramatic age-dependent change in the cyclic AMP-stimulated phosphorylation of endogenous proteins of molecular weights 73,000 (1a), 68,000 (1b), 65,000 (1c) and 55,000. Experiments in which isolated SJs were phosphorylated with soluble cyclic AMP-dependent kinase showed that the phosphorylation of proteins 1a, 1b and 1c resulted from their de novo appearance in newborn SJ fractions and not from a maturation-dependent coupling of kinases and substrates that were already present in newborn SJ fractions. The levels of regulatory (R) subunits of cyclic AMP-dependent kinases in synaptic fractions were measured by [3H]cyclic AMP binding and photoaffinity labeling with [32P]8-N3 cyclic AMP and were observed to change little during postnatal development. These findings show that there is a strong correlation between the in situ appearance of synapses and the enzymatic maturation of endogenous, cyclic AMP-stimulated protein phosphorylation in SJ fractions isolated at different stages of development. PMID- 6289985 TI - Persistent effects of amphetamine on cerebellar Purkinje neurons following chronic administration. AB - The spontaneous discharge of cerebellar Purkinje neurons was studied in rats after withdrawal from chronic treatment with amphetamine (2 mg/kg per day x 21 days). Discharge rates in withdrawn animals remained significantly lower than those of controls for up to 50 days. Disruption of the adrenergic input to these neurons from the locus coeruleus by treatment with propranolol, clonidine or reserpine, partially restored these discharge rates. Acute administration of amphetamine in amphetamine-withdrawn rats did not further depress Purkinje neurons discharge rate, whereas in a previous study in this lab, Purkinje neurons from naive animals were markedly slowed. Moreover, Purkinje neurons from amphetamine-withdrawn rats were also significantly less sensitive than controls to locally applied norepinephrine. These results demonstrate that chronic amphetamine can lead to very long-term changes in neuronal activity, and suggest that these changes may be mediated, in part, by the noradrenergic transmitter systems. PMID- 6289986 TI - Effects of intrathecal ACTH on opiate analgesia in the rat. PMID- 6289987 TI - Neurotransmitter receptor sites in human hippocampus: a quantitative autoradiographic study. PMID- 6289988 TI - Alterations of membrane integrity and cellular constituents by arachidonic acid in neuroblastoma and glioma cells. AB - Effects of arachidonic acid on cellular metabolism, cation content, lipid peroxidation, sodium pump activities and release of labeled arachidonic acid were studied in C-6 glioma cells and N18TG2 neuroblastoma cells. Arachidonic acid caused a significant increase in intracellular sodium levels concomitant with a decrease in intracellular potassium in both cell lines. Both (Na+ + K+)-ATPase and p-nitrophenyl phosphatase of glioma cells were inhibited by arachidonic acid whereas only the p-nitrophenyl phosphatase of neuroblastoma cell was inactivated. Low concentrations of arachidonic acid stimulated lactic acid release whereas high concentrations had an opposite effect. In addition, the lipid peroxide content of glioma cells was increased abruptly by 50 microM arachidonic acid whereas only a slight increase of malondialdehyde was observed in neuroblastoma cells. When the cultured cells of both cell lines were incubated with exogenous labeled arachidonic acid, 78-95% of the label was incorporated into membrane phospholipids. Only a very small fraction of prostaglandin E2 and prostaglandin F2 alpha was synthesized. Exogenous arachidonic acid and free radicals generated with xanthine-xanthine oxidase caused a significant release of endogenous labeled arachidonic acid from cellular membrane phospholipids. These data further support our hypothesis that the arachidonic acid and its oxygen radical metabolites induce pathological alterations in membrane permeability and cellular volume. PMID- 6289989 TI - Striatal lesions change the behavioral effects of morphine in cats. AB - Cats injected with a relatively low single dose of morphine sulfate (0.5-3.0 mg/kg i.p.) exhibit a long-lasting group of behaviors which we quantified via a time-sampling video technique. The dominant events are complex head movements accompanied by discrete paw, ear and body movements with the animal in a quiet posture, all of which appeared to be visually mediated. Cats with extensive lesions of the caudate nuclei do not show this profile; instead they show unspecific locomotor activity proportional to the size of the ablation and to the dose of morphine. These effects are blocked by naloxone in both intact and lesioned animals. The robustness of these results indicate that (i) the striatum is involved in the behavioral effects of morphine, and (ii) that the cat is a useful, sensitive model for the study of the behavioral effects of opiates. PMID- 6289990 TI - Effects of ACTH4-10 on vestibular compensation. AB - ACTH4-10, a fragment of the adrenocorticotropic hormone (ACTH) molecule, has marked effects on the compensation process following unilateral labyrinthectomy. In Rana temporaria ACTH4-10-treatment (5-250 micrograms/kg) influences both the acquisition and the maintenance of the compensated state. The compensation process is slowed down by hypophysectomy but can then be restored by the administration of ACTH4-10. It is concluded that ACTH-like neuropeptides might physiologically be involved in the plastic processes underlying functional recovery from CNS lesions. PMID- 6289991 TI - The effect of castration on stria terminalis neurone absolute refractory periods using different antidromic stimulation loci. PMID- 6289993 TI - Opiate and opioid peptide binding in rat goldfish: further evidence for opiate receptor heterogeneity. AB - The binding of a series of 3H-labeled mu, kappa, sigma and delta opioid agonists and an antagonist has been examined in rat and goldfish brain membranes. A variety of treatments, including NaCl, N-ethylmaleimide, trypsin and chymotrypsin show dramatic differences both between the two species and between the different 3H-ligands. PMID- 6289992 TI - Effect of transection of subfornical organ efferent projections on vasopressin release induced by angiotensin or isoprenaline in the rat. AB - Transection of subfornical organ efferents in the rat prevented the vasopressin release in response to intravenous angiotensin II infusion or following a small dose of the beta-sympathomimetic amine isoprenaline (30 micrograms/kg i.m.). In contrast, this lesion had no effect on vasopressin release after hypertonic saline injection or a high dose of isoprenaline (480 micrograms/kg i.m.). We conclude that blood-borne angiotensin II induces vasopressin release by acting on the subfornical organ; depending on the dose of isoprenaline, activation of the endogenous renin-angiotensin system may mediate isoprenaline-induced vasopressin release. PMID- 6289994 TI - Muscarinic receptors on dopamine terminals in the cat caudate nucleus: neuromodulation of [3H]dopamine release in vitro by endogenous acetylcholine. AB - The directly acting muscarinic receptor agonist oxotremorine (1.8-10 microM) produced an increase in electrically evoked [3H] dopamine release from slices of the cat caudate. The maximal increase caused by oxotremorine was 40%, and was antagonized by the muscarinic receptor blocking agent atropine (0.1 microM). Exposure to the acetylcholinesterase (AChe) inhibitor physostigmine (1 microM) resulted in a 50% increase in electrically evoked [3H]dopamine release. The increase caused by physostigmine was also antagonized by atropine (0.1 microM). Atropine did not, however, alter the modulations in [3H]dopamine release mediated by the dopamine autoreceptor: the increase in electrically evoked [3H]dopamine release caused by the dopamine receptor antagonist S-sulpiride (0.1 microM) and the decrease caused by the dopamine receptor agonist pergolide (30 nM) were unaffected by atropine (0.1 microM). These results indicate that the muscarinic receptor-mediated and dopamine autoreceptor-mediated presynaptic effects on [3H]dopamine release are independent. The present results suggest that in the electrically depolarized caudate slice in vitro, released endogenous acetylcholine may interact with muscarinic receptors facilitating depolarization evoked [3H]dopamine release, if AChE is inhibited. These muscarinic receptors may be located on dopamine nerve terminals. In the context of present neuroanatomical knowledge, the action of released endogenous acetylcholine on dopamine terminals may be a non-synaptic neuromodulation. PMID- 6289995 TI - Medullary ventral surface GABA receptors affect respiratory and cardiovascular function. AB - We previously demonstrated that GABA and muscimol administered either into the cisterna magna or the fourth ventricle to chloralose-anesthetized cats cause respiratory depression, hypotension, and bradycardia. Injection of these substances into the lateral and third ventricles had no effect. In order to localize the site of action, muscimol and GABA were applied by Perspex rings to the ventral surface of the medulla. Application of muscimol (0.25-2.66 micrograms) to Schlaefke's area in 6 cats reduced minute ventilation from 443 +/- 38 to 291 +/- 52 ml/min by reducing tidal volume from 31.8 +/- 2.3 to 17.6 +/- 1.4 ml, without changing respiratory rate and duration of inspiration. Hypotension and bradycardia were also observed. Application of GABA (0.14-4.86 mg) produced similar effects on respiratory activity and arterial blood pressure. No significant effects occurred when high doses of these agents were applied to Loeschcke's and Mitchell's areas. Application of bicuculline (5-25 micrograms) to Schlaefke's area had the opposite effect of muscimol and GABA on respiratory activity and blood pressure, and reversed the respiratory and cardiovascular depressant effects of both agents. We conclude that GABA receptors are present at Schlaefke's area, and that activation of these receptors results in respiratory depression, hypotension, and bradycardia. Our results suggest that GABA may be an important inhibitory neurotransmitter in the modulation of respiratory and cardiovascular control. PMID- 6289996 TI - Diethylstilbestrol regulates the number of alpha- and beta-adrenergic binding sites in incubated hypothalamus and amygdala. AB - Previous work has identified an effect of circulating estrogens on the number of central adrenergic binding sites. We have further characterized this effect by performing the experiments in vitro and have taken advantage of a well-described hypothalamic preparation in which diethylstilbestrol (DES), is known to elevate cAMP levels through a pathway which involves adrenoreceptors. We show that DES induces a reciprocal change in the numbers of alpha- and beta-adrenergic binding sites in incubated hypothalami obtained from intact immature female rats as well as from ovariectomized adult rats. The alpha-adrenergic binding sites are reduced by 25-30% whereas the beta-adrenergic sites are increased by 60-100%. The effect is maximal at 3 h in vitro (20 microM DES) and largely reversible following a 2 h wash in the absence of DES. Using the change in beta-adrenergic binding sites as a probe, we were further able to show that estradiol (100 microM) and 2 hydroxyestradiol (50 microM) had no effect. Further, the effect of DES was not blocked by the anti-estrogens clomiphene or tamoxifen. Since DES is able to elevate beta-adrenergic binding sites in hypothalamus and amygdala (brain areas known to contain high levels of estrogen receptors) but has no effect in cerebellum, we conclude that we have observed an effect of DES not shared by estradiol but which may be confined to estrogen target areas of the brain. PMID- 6289997 TI - Opiate receptor binding sites in human brain. AB - Subclasses of opiate receptor binding sites in human brain membranes were investigated by means of competitive binding techniques. The experimental data were analyzed by use of a computerized non-linear regression curve fitting program. mu-, delta-and chi-types of opiate binding were found in 5 different regions of the brain. A more extensive analysis of the regional distribution of subclasses of opiate binding sites was performed using a simple sequential inhibition technique. This method was shown to yield results which are comparable to those obtained by computer analysis of multiple tracer displacement curves. Chi-and mu-sites represented the major component of binding in most brain areas whereas delta-sites were fewer in number. The 3 types of binding showed different distribution patterns, suggesting that they are independent from each other. The distribution pattern observed in human brain resembled the one observed in rat brain, although chi-sites appear to represent a more important, and delta-sites appear to represent a less important, fraction of binding in human as compared to rat brain. PMID- 6289998 TI - Alterations in the ipsi- and contralateral afferent inputs of dorsal horn cells produced by capsaicin treatment of one sciatic nerve in the rat. AB - Dorsal horn cells in the lumbar spinal cord of decerebrate spinal rats were examined 7-21 days following local application of capsaicin to the sciatic nerve. Such local capsaicin treatment is known not to influence the size of the incoming A and C fibre afferent volley. The receptive field properties and primary afferent input of cells on both sides of the cord, that is ipsi and contralateral to the treated nerve, were examined. On the treated side, the percentage of cells excited by C fibres from the capsaicin treated nerve was 30% of normal and the number of cells responding to noxious heating of the cutaneous receptive field was reduced by 50%. A fibre input and low and high threshold mechanical input were normal. The receptive field size was larger in many cells innervated by the treated nerve. On the side opposite to the treated nerve, responses to noxious and non-noxious stimulation of the untreated limb were unaffected as was the input from the untreated sciatic nerve. Receptive fields were somewhat larger than normal. Effects were also observed from contralateral stimuli. Cells on both sides of the cord were found with excitatory contralateral receptive fields and excitatory responses to trains of high intensity stimulation to the contralateral sciatic nerve. In untreated animals the effect of such contralateral stimulation is inhibitory. The results show that peripheral nerve capsaicin treatment causes long lasting reduction of the C fibre input to dorsal horn cells on the treated side. However, it also results in changes in the inhibitory and excitatory receptive fields of cells on both sides of the cord. PMID- 6289999 TI - Effect of serum-free medium on growth and differentiation of sympathetic neurons in culture. AB - Primary cultures of dissociated superior cervical ganglia (SCG) were grown either in serum-containing medium (+FBS) or in serum-free, defined medium (N1). The cultures were then compared for several differentiated properties. Neuronal survival was similar in the two media. Background cell, especially fibroblast, proliferation was less in N1. Small, intensely fluorescent cells were occasionally seen only in +FBS. Catecholamine fluorescence in neuronal processes and cholinergic synaptic activity persisted beyond 1 month in culture in both media. Quantitatively, however, neuritic outgrowth, nerve terminal fluorescence, and synaptic interaction were less in N1. Muscarinic depolarization and electrical membrane properties, including the presence of at least 4 voltage sensitive outward currents, were similar in the two media. Thus, SCG neurons differentiate in N1 essentially to the same degree as they do in +FBS. PMID- 6290000 TI - Fetally-induced noradrenergic hyperinnervation of cerebral cortex results in persistent down-regulation of beta-receptors. AB - Methylazoxymethanol acetate (MAM)-induced cortical hypoplasia resulted in a 2 fold increase in the concentration of norepinephrine and its metabolite, 3 methoxyl-4-hydroxyphenylglycol sulfate, and a 26% decrease in the Bmax for the beta-receptor ligand, [3H]dihydroalprenolol (DHA) in the rat cortex. Chronic treatment with desmethylimipramine did not further decrease DHA-labeled sites although prior lesion of the noradrenergic terminals with 6-hydroxydopamine markedly increased the number in the MAM-lesioned cortex. PMID- 6290001 TI - [Arterial, venous and lymphatic components of liver cancer microcirculation]. PMID- 6290002 TI - [Probable inhibition of a bacterial collagenase by serotonin]. AB - We have previously demonstrated that intraperitoneal injections in rats of collagenase (from Clostridium histolyticum) are followed by severe lesions. Here we show that the injection of collagenase together with serotonin has no or only small effects. It appears to be partly due to serotonin. Creatinin and sulfate which form an injectable complex with serotonin have no influence on this phenomenon. The part of pH 4,4 of the injected solution is discussed. These facts suggest that serotonin may perhaps lower the collagenolysis and be thus responsible for the fibrogenetic effects of carcinoid tumors, many of these being characterised by their high production of serotonin. PMID- 6290003 TI - [Urinary excretion of calcium and cyclic adenosine monophosphate in children. Comparison between the two variables]. AB - The urinary excretion of calcium (mg/g creatinine) and cyclic adenosine 3'5' monophosphate (cyclic AMP) (mumol/g creatinine) was determined in 79 healthy children aged 2 to 12 years. An ag--related pattern to the urinary excretion of calcium was not seen. But urinary cyclic AMP levels are inversely correlated with age (r = -0,66; p less than 0,01) and there is a significant correlation between urinary calcium and cAMP levels (r = 0,2; p less than 0,05). PMID- 6290004 TI - [Common proviral integration sites in C57BL mouse lymphoma induced by mouse radiation leukemia virus and absence of novel proviral sequences in DNA from radiation-induced lymphoma]. AB - Radiation leukemia virus (RadLV)-induced lymphomas of C57BL mice display the appearance of novel proviral sequences at common sites of their DNAs. These results are compatible with the hypothesis of viral oncogenesis by promotor insertion. Radiation-induced tumors do not acquire detectable novel provirus, suggesting that activation of a transforming gene proceeds by a mechanism distinct from the former. PMID- 6290005 TI - [Relationship between hepatocellular carcinoma, liver cirrhosis and hepatitis B: a pathological study]. PMID- 6290006 TI - [Isolation of cytomegalovirus]. PMID- 6290007 TI - [Detection of IgM and IgG antibodies to hepatitis A virus by reversed passive hemagglutination inhibition test]. PMID- 6290008 TI - Comparison of complexes containing lysyl-tRNA synthetase from normal and virus transformed cells. AB - We compared the lysyl-tRNA synthetases from normal (Balb/3T3) and murine sarcoma virus-transformed (KA31) mouse fibroblasts. In agreement with several other reports of mammalian systems, the lysyl-tRNA synthetases from these cells occurred in very large postmicrosomal complexes as determined by gel filtration on agarose columns. Arginyl-, isoleucyl-, methionyl-, phenylalanyl-, and tyrosyl tRNA synthetases also occurred as part of a large complex or complexes. Activity of glycyl- or leucyl-tRNA synthetase was not detected in a complex. The specific activities of arginyl- and methionyl-tRNA synthetases were three- and five-fold higher, respectively, in a complex from KA31 as compared with a complex from Balb/3T3. In contrast, the specific activity of lysyl-tRNA synthetase from the Balb/3T3 complex was 50% higher than that of the KA31 complex. tRNALys obtained from the complexes of Balb/3T3 and KA31 was fractionated into isoacceptors on columns of RPC-5. The relative amounts of lysine isoacceptors in total preparations of tRNA from normal whole cells and in tRNA obtained from the normal enzyme complex were the same. However, two isoacceptors were present in greater amounts and two were present in lesser amounts in the KA31 enzyme complex as compared with lysine isoacceptors in a total preparation of tRNA from KA31 cells. PMID- 6290009 TI - ESR studies on the lipid bilayers of separated outer and cytoplasmic membranes of a moderately halophilic bacterium. PMID- 6290010 TI - Tonin as activator of renin. AB - Tonin, a new serine protease, found in high concentration in rat submaxillary glands, leads to a significant activation of human amniotic fluid renin. The optimum pH on renin activation by tonin was found at pH 6.0. The reaction was time dependent and the initial rate of angiotensin I generation was constant up to 2 h. The two amniontic fluid samples studied showed an increase in renin activity after incubation with tonin to about five times the control level (268 to 1240 pmol x h-1 x mL-1 and 1490 to 7480 pmol x h-1 x mL-1). PMID- 6290012 TI - Immune responses of dairy cattle to parainfluenza-3 virus in intranasal infectious bovine rhinotracheitis-parainfluenza-3 virus vaccines. AB - Two hundred and fifty dairy heifers were vaccinated at three to six months of age with an intranasal infectious bovine rhinotracheitis-parainfluenza-3 vaccine. Eighteen additional heifers were tested prior to vaccination and again three to four weeks after vaccination. Neither cell-mediated nor humoral immunity was significantly raised to parainfluenza-3 virus in either group of cattle. PMID- 6290013 TI - Fall in antibody titer to bovine leukemia virus in the periparturient period. AB - Twenty-seven cows with antibodies to bovine leukemia virus were bled before, during and after calving. All serum samples were tested quantitatively for bovine leukemia virus antibodies using both the agar-gel immunodiffusion test with a glycoprotein antigen and the radioimmunoprecipitation assay with an internal p24 protein antigen. A significant fall (P less than 0.001) in bovine leukemia virus antibody titer was demonstrated with both tests at the time of calving, with a subsequent rise in antibody titer within one month of parturition. Bovine leukemia virus antibodies were not detectable using the agar-gel immunodiffusion test in two of these cows at the time of calving. PMID- 6290014 TI - Aerosol vaccination of calves with pasteurella haemolytica against experimental respiratory disease. AB - Three experiments were conducted on calves in which the efficacy of vaccination with live Pasteurella haemolytica in aerosol was tested by challenge with sequential aerosol exposure to bovine herpesvirus 1 and P. haemolytica. Neither single nor multiple aerosol vaccinations protected against the experimental disease. Macroscopically recognizable rhinitis, tonsillitis, tracheitis and pneumonia occurred in both controls and vaccinates. In one experiment as many as three aerosol vaccinations with live P. haemolytica for up to 20 minutes failed to elicit clinical signs in exposed calves. Pasteurella haemolytica was isolated less frequently from tissues of vaccinated calves than from those of nonvaccinated calves. Pasteurella haemolytica was isolated from deep nasal swabs of 4/14 vaccinated calves five and six days after viral exposure. It was concluded that although bovine herpesvirus 1 vaccination has been shown previously to prevent the experimental disease produced by bovine herpesvirus 1 P. haemolytica, live P. haemolytica vaccination by aerosol will not provide the same protection. PMID- 6290016 TI - Oxytocin action: lipid metabolism in adipocytes from homozygous diabetes insipidus rats (Brattleboro strain). AB - Oxytocin, like insulin, stimulates glucose oxidation in normal rat adipocytes. Fat cells from homozygous Brattleboro rats that exhibit diabetes insipidus (HoDI animals) and that have a normal number of oxytocin receptors, however, are unable to respond to oxytocin in terms of glucose oxidation. We now report that in adipocytes from HoDI animals that are responsive to insulin, oxytocin was also unable to stimulate lipogenesis. In contrast, oxytocin like insulin was able to inhibit epinephrine-stimulated lipolysis in adipocytes from HoDI animals. Thus, in HoDI adipocytes, the results indicate that the receptor-effector system is only partially defective. PMID- 6290015 TI - Respiratory disease in calves produced with aerosols of parainfluenza-3 virus and Pasteurella haemolytica. AB - In four experiments, 22 calves were exposed to aerosols of parainfluenza-3 virus, followed by Pasteurella haemolytica at intervals of three to 13 days. The purpose of each experiment was to study viral-bacterial interactions in the respiratory tracts. Two experiments, in which the viral aerosols were diluted by the addition of air, produced sporadic temperature elevations while two experiments with undiluted viral aerosols produced consistent temperature elevations. Diluted viral aerosols produced lobular sized lesions in the lungs and hemagglutinating inhibition antibodies in sera, whilst undiluted aerosols produced a synergistic effect in the form of purulent pneumonia in ten of 14 calves when the interval between viral and bacterial aerosols was from three to ten days. Histopathological changes attributable to the virus only were seen in all experiments, and the histopathological changes due to mixed infection of parainfluenza-3 virus and P. haemolytica are described in detail. This is the first report of extensive purulent pneumonia in calves after parainfluenza-3 virus and P. haemolytica exposure. This was achieved using much smaller inocula than in experiments previously reported. PMID- 6290017 TI - Nonspecific proteases: a new approach to the isolation of adult cardiocytes. AB - The potent collagenolytic activity of nonspecific proteinases suggested their use as enzymatic agents for the dissociation of single adult heart cells. This was assessed in guinea pig hearts perfused for 1 min with solutions containing hyaluronidase (100-10 000 U/mL), trypsin (100-10 000 U/mL), crude collagenase (100-500 U/mL), or nonspecific protease (0.1-100 U/mL). No rod-shaped cells were observed among the cells isolated with these concentrations of hyaluronidase, trypsin, or crude collagenase. By contrast, 45-80% of the cells released with nonspecific protease (5-10 U/mL) were rod shaped and Ca2+ tolerant. Resting and action potentials recorded from cells dispersed with nonspecific protease were similar to those recorded from cells isolated after prolonged collagenase exposure. PMID- 6290018 TI - Antagonism with dibenamine, D-600, and Ro 3-7894 to estimate dissociation constants and receptor reserves for cardiac adrenoceptors in isolated rabbit papillary muscles. AB - In papillary muscles isolated from reserpinized rabbits, positive inotropic responses to the alpha (alpha)-adrenergic agonist, (-)-phenylephrine in the presence of 10(7) M timolol and the beta (beta)-adrenergic agonist. (-) isoproterenol were antagonized with the irreversible alpha-adrenergic antagonist, dibenamine, the irreversible beta-adrenergic antagonist. Ro 3-7894, and the calcium blocker, D-600. D-600 was employed as a functional antagonist of both alpha- and beta-adrenoceptor responses. Dissociation constants (Ka values) for drug-receptor interactions were calculated by the method of Furchgott and used to estimate fractional receptor occupancy and agonist efficacies. Comparison of responses showed that the receptor reserve for cardiac beta-adrenoceptors was greater than for alpha-adrenoceptors. D-600 was an effective inhibitor of both cardiac alpha- and beta-adrenoceptor responses; however, estimates of KA and receptor reserves were similar to estimates using an irreversible antagonist for alpha-but not beta-adrenoceptors. PMID- 6290019 TI - Beta 2-mediated hypotension and myocardial injury. AB - This study was designed to investigate the importance of beta 2 receptor mediated hypotension in the pathogenesis of myocardial injury. The effect of isoproterenol and the putative beta 2 agonist albuterol on arterial blood pressure, heart rate, the myocardial content of ATP and cAMP, and the serum content of MB-CPK was examined in conscious rats. Isoproterenol (5.25 mg/kg, s.c.) and albuterol (45 mg/kg, s.c.) lowered blood pressure and elevated heart rate to the same extent. Also, both agonists increased the myocardial content of cAMP, decreased the myocardial content of ATP, and elevated serum MB-CPK. The beta 1 antagonist practolol, but not the ganglionic blocking agent chlorisondamine, attenuated the elevation in heart rate to albuterol without reducing its effect on blood pressure. Practolol, but not chlorisondamine, abolished the effects of albuterol on cAMP, ATP, and MB-CPK. These data suggest that the myocardial injury which is associated with an increased heart rate and changes in cAMP, ATP, and MB-CPK following the administration of albuterol is not the result of beta 2-mediated hypotension, but is due to stimulation of myocardial beta 1 receptors. PMID- 6290011 TI - A review of infectious bovine rhinotracheitis, shipping fever pneumonia and viral bacterial synergism in respiratory disease of cattle. AB - Unanswered questions on the etiology and prevention of shipping fever pneumonia have allowed this disease to remain one of the most costly to the North American cattle industry. Research in this area has indirected that while Pasteurella haemolytica and, to a lesser extent, P. multocida are involved in most cases, they seem to require additional factors to help initiate the disease process. Bovine herpes virus 1 has been shown experimentally to be one such factor. This review examines in some detail the topics of infectious bovine rhinotracheitis, shipping fever, and viral-bacterial interactions in the production of respiratory disease in various species. It deals with history, definitions, etiologies, clinical signs and lesions, and considers exposure levels, transmission and various pathogenetic mechanisms that are postulated or known to occur. PMID- 6290020 TI - Methotrexate with citrovorum factor rescue as primary therapy for gestational trophoblastic disease. AB - Methotrexate with citrovorum rescue (MTX-CF) was administered as primary treatment in 106 patients with gestational trophoblastic disease (GTD). Ninety six patients (90.6%) achieved complete remission with MTX-CF and 77 of these patients (80.2%) required only one course of MTX-CF to attain remission. MTX-CF induced sustained remission in 89 (94.7%) of 94 patients with nonmetastatic GTD and in seven (59.3%) of 12 patients with low-risk metastatic GTD. Resistance to MTX-CF was more common in patients with disseminated disease and with pretreatment hCG titers greater than or equal to 50,000 milliIU/ml. Following MTX CF, granulocytopenia, thrombocytopenia and hepatotoxicity was observed in only seven (6.6%), three (2.8%), and ten (9.4%) patients, respectively. MTX-CF should be the preferred primary treatment in nonmetastatic and low-risk metastatic GTD. PMID- 6290022 TI - A comparison of World Health Organization (WHO) classification of lung tumors by light and electron microscopy. AB - Forty-nine cases of lung tumor were initially classified according to the World Health Organization (WHO) system by light microscopy. Additional study by electron microscopy using only a minimum of ultrastructural features as criteria for each type made more precise classification of some of the poorly differentiated or undifferentiated tumors possible. Most significant was the finding of evidence of a second tumor type in nine of the 35 tumors previously classified as moderately to well differentiated thus reclassifying them as mixed tumors. Sixteen of the 49 tumors were reclassified and a more precise classification was made in five others. Only two cases of undifferentiated small cell tumors were available and no unequivocal neurosecretory granules could be found in either. Correlation of the more precise typing of lung tumors by electron microscopy with the clinical data and modes of therapy will provide necessary information for determining the usefulness of the WHO classification. PMID- 6290021 TI - Experimental studies on malignant fibrous histiocytomas in rats. I. Production of malignant fibrous histiocytomas by 4-hydroxyaminoquinoline 1-oxide in bone of Fischer 344 strain rats. AB - Malignant fibrous histocytomas (MFHs) were induced by a single injection of 4 hydroxyaminoquinoline 1-oxide (4-HAQO) into periosseous tissue of the tibia or by inserting solid 4-HAQO into the bone marrow of the tibia of male Fischer 344 rats. Periosseous MFHs were induced by doses of 2 mg and 4 mg of 4-HAQO per rat in three of 13 (23%) rats and nine of 13 (69%) rats, respectively, at 18 to 27 weeks after treatment. Bone MFHs were induced by doses of 2 mg, 4 mg, and 8 mg of 4-HAQO per rat in one of 15 (7%) rats, 11 of 18 (61%) rats, and 12 of 14 (86%) rats, respectively, after 18 to 29 weeks. Radiologic examination of bone MFH revealed bone destruction with or without a periosteal reaction, pathologic fracture, and tumor invasion into periosseous soft tissue. The serum alkaline phosphatase levels were elevated in rats with bone MFHs. Histologically, these MFHs were divided into fibrous, giant cell, myxoid, and inflammatory types. The incidence of fibrous MFHs was highest. Lung and inguinal and/or retroperitoneal lymph node metastases were observed in some rats, and these were of the fibrous type. These MFHs were quite similar to those in humans histologically and electron microscopically. This work establishes an animal model for studying the pathogenesis of bone MFH. PMID- 6290023 TI - The frequency and clinical biology of the ectopic hormone syndromes of small cell carcinoma. AB - Eighty-four patients with small cell carcinoma of the lung were reviewed with respect to the frequency and biologic behavior of patients with a demonstrated ectopic hormone syndrome. This subgroup of 12 patients (14%) was compared with regard to stage of disease, distribution of metastases, response rate, and survival to those patients not demonstrating a clinical syndrome. Stage, distribution, and response rates are comparable in the two groups but differences were observed with regard to sites of metastases and survival. The ectopic hormone syndrome patients had an increased likelihood of liver metastasis and CNS metastasis. The frequency of CNS metastasis in patients without the ectopic hormone syndrome was 11/72 (15%) of which four of the 11 were present at initial presentation. In contrast, 5/12 (42%) of the patients with ectopic hormone syndromes developed CNS metastasis, and all five had clinically demonstrated SIADH. Survival was consistently inferior in those patients with the ectopic hormone syndrome. PMID- 6290024 TI - The pathological findings of breast cancer in patients surviving 25 years after radical mastectomy. AB - The authors report the findings in 107 women who are known to have survived 25 years from among a population of 746 consecutive patients who underwent radical mastectomy for breast carcinoma at the University of Chicago Hospitals and Clinics from 1929 to 1955. Of these patients, 103 had invasive carcinomas, two had intraductal carcinomas, and two had subareolar papillomatosis. Six patients had to be excluded because of inadequate pathologic material. The pathologic findings in 93 cases were compared with those in an equal number of control cases dying within a comparatively short period (median, 3.4 years; range 0.9-9.9 years) after radical mastectomy. These were matched for age, tumor size, and number of positive nodes. Only two of our patients suffered recurrences, and none died of her original tumor; however, 12 developed second primaries in the opposite breast, and four died from them. Compared with all patients who underwent radical mastectomy in this period, the 25-year survivors were younger (69 versus 43% were younger than age 50 years), had smaller tumors (39 versus 26% less than 2 cm in diameter), and a larger number (60 versus 39%) had negative nodes. Nonetheless, 12% of the survivors had tumors larger than 5 cm in diameter and 11% had four or more positive nodes. Histologically, 19% of the 25-year survivors had medullary, mucoid, infiltrating lobular, tubular or lipid rich carcinomas, whereas there was only one lobular and one apocrine carcinoma in the control group. Compared with controls, the survivors had a higher percentage of Grade I tumors and a lower incidence of lymphatic and vascular invasion in the breast. Only one 25-year survivor compared with 16 controls had blood vessel invasion. A surprising 63% of the 25-year survivors had lymphatic or vascular invasion within the tumor, or lymph node metastases compared with 82% of controls. While our studies confirm the importance of these well-known prognostic indicators, it also shows that some patients with pathologically unfavorable lesions, i.e., large tumors of high grade with extensive lymphatic invasion and many positive nodes, treated by radical mastectomy may survive for 25 years. However, we could not accurately predict, among the cases we studied, who would be expected to survive 25 years or who would die within four years. PMID- 6290025 TI - Endocrine function in small cell undifferentiated carcinoma of the lung. AB - The endocrine status of 106 patients with undifferentiated small cell carcinoma of the lung was evaluated before treatment was begun. Almost one half of the patients had evidence of abnormal control of the secretion of adrenal cortical steroids, manifested by loss of diurnal rhythmicity or dexamethasone suppressibility. Only two had the clinical syndrome of ectopic ACTH secretion. Evidence of inappropriate secretion of vasopressin was found in 38% of the patients, most of whom also had abnormalities of corticosteroid secretory pattern. About one half of the patients had evidence of abnormal glucose tolerance, and many also had a paradoxical rise of plasma growth hormone concentration after glucose administration. The levels of the other hormones studies were normal. The pattern of hormone abnormality observed in these patients appears to be relatively specific for small cell undifferentiated carcinoma, and is different from that observed in other pulmonary tumors. Patients with abnormal control of plasma cortisol had a worse prognosis than those with normal adrenal function, largely because of decreased response rates to chemotherapy. Other endocrine abnormalities were of no prognostic significance. PMID- 6290027 TI - Paget's disease of the skin. A unifying concept of histogenesis. AB - Utilizing the peroxidase antiperoxidase technique we explored the presence and the distribution of carcinoembryonic antigen (CEA) in seven cases of mammary and sixteen of extramammary Paget's disease of skin. In every case positive immunostaining was observed in all the Paget's cells and the underlying tumor (where one was present), whereas intervening keratinocytes and melanocytes did not stain. CEA was also present in the cells and secretions of normal eccrine and apocrine glands. Our observations confirm that Paget's cells are of glandular origin. PMID- 6290026 TI - Plasma CEA levels in small cell lung cancer. Correlation with stage, distribution of metastases, and survival. AB - Plasma CEA levels were determined in 61 patients with small cell lung cancer entering chemotherapy protocols between 1976 and 1980. Five quantitative categories were determined: less than 2.5 ng/ml (the standard normal for Hansen assay); 2.6-5.0 ng/ml (the extended normal for smokers); 5.1-20.0 ml; (the intermediate elevation and the transition for the indirect to the direct assay); 20-100 ng/ml; and greater than 100 ng/ml. Forty-seven percent of patients had levels less than 5 ng/ml and only ten of 61 or 16% had levels greater than 20 ng/ml. There was no clearcut relationship of plasma CEA level to stage of disease, in that 40% of patients with extensive disease (32 patients) had levels less than 5 ng/ml and 45% of patients with limited disease (29 patients) had levels greater than 5 ng/ml. Similarly, there was no relationship of CEA level to site of metastases, although levels greater than 100 ng/ml were always associated with liver metastases. The median survival for each quantitative category was similar, ranging from seven to nine months. The use of sequential determinations of CEA to correlate with tumor response was studied in only eight patients with levels greater than 20 ng/ml and a measurable parameter of disease. The qualitative direction of change of CEA was appropriate in those patients responding to treatment but in three nonresponding patients the direction of CEA change paradoxically decreased. In five patients who developed progressive disease, the plasma CEA level predicted the clinical relapse. CEA as a tumor marker for oat cell carcinoma must be applied in conjunction with other tumor parameters and is meaningful in only a small proportion of the total small cell patient population. PMID- 6290028 TI - Activation of Epstein-Barr virus expression in human lymphoblastoid P3HR-1 and Raji cells with propionic acid and with culture fluids of propionic acid producing anaerobes. AB - Epstein-Barr virus (EBV)-associated early (EA) and virus capsid antigens (VCA) were efficiently induced in the viral genome-carrying human lymphoblastoid cells, P3HR-1 and Raji, by the culture fluids of Propionibacterium acnes, P. avidum, P. lymphophilum and Arachnia propionica, the anaerobes which are commonly seen among the normal flora of man. The active principle for EBV-induction in the 2 cell lines was the propionic acid produced by the microbes and such activity was shown to correlate with the fatty acid content of the culture media. PMID- 6290029 TI - Simple sandwich enzyme immunoassay for quantification of mouse mammary tumor virus in mouse milk. AB - We have developed a sandwich enzyme immunoassay in order to measure quantitatively mouse mammary tumor virus (MMTV) in mouse milk. In this assay, the antibody-beta-D-galactosidase complex and antibody-bound silicon rubber pieces pieces as solid phase are used. The assay is able to detect 10 ng/ml of MMTV in the milk sample. PMID- 6290031 TI - Heterogeneity of expression and induction of mouse mammary tumor virus antigens in mouse mammary tumors. AB - Seven spontaneous BALB/cfC3H mouse mammary tumors were heterogeneous in expression of murine mammary tumor virus-associated cell surface antigens. To determine the basis of this heterogeneity, cells from spontaneous tumors and from five subpopulations isolated from a single spontaneous tumor were examined for expression of viral antigens under both conventional conditions and conditions known to induce synthesis of murine mammary tumor virus antigens (5-iodo-2' deoxyuridine and dexamethasone). Induction with 5-iodo-2'-deoxyuridine resulted in further manifestation of the antigenic heterogeneity of spontaneous tumors. The five subpopulations from a single tumor differed in the amount of viral antigens present in untreated and in induced cultures. Coculturing showed that viral antigen expression was independent in each subpopulation within a heterogeneous mixture and was not influenced by the presence of other subpopulations with different potentials for viral antigen synthesis. The expression of murine mammary tumor virus structural antigens, a protein with a molecular weight of 28,000 and a glycoprotein with a molecular weight of 52,000, differed within the heterogeneous subpopulations, and was noncoordinate. The data suggest that the antigenic heterogeneity in spontaneous tumors reflects the existence of cells within them that differ in both expression of viral antigens and in their response to inducers of viral antigen synthesis. PMID- 6290030 TI - Characterization of an adenosine 5'-triphosphate- and deoxyadenosine 5' triphosphate-activated nucleotidase from human malignant lymphocytes. AB - The kinetic properties of a soluble, magnesium-dependent 5'-nucleotidase from human malignant lymphocytes have been determined. The partially purified enzyme is distinct from plasma membrane-associated 5'-nucleotidase and is free of nonspecific phosphatase activity. Among purine ribonucleotides, it reacted efficiently with inosine 5'-monophosphate and guanosine 5 -monophosphate and to a lesser degree with deoxyguanosine 5'-monophosphate. Adenosine 5'-monophosphate and deoxyadenosine 5'-monophosphate were 30-fold less efficient substrates. Increasing concentrations of adenosine 5'-triphosphate and deoxyadenosine 5' triphosphate from 0 to 3 mM enhanced 5'-nucleotidase activity up to 7-fold. Guanosine 5'-triphosphate and deoxyguanosine 5'-triphosphate were much less effective enzyme activators, while uridine 5'-triphosphate was without effect. Inorganic phosphate inhibited dephosphorylating activity in both adenosine 5' triphosphate-supplemented and unsupplemented buffer. The activation of this 5' nucleotidase by deoxyadenosine 5'-triphosphate, combined with the relative inability of the enzyme to dephosphorylate deoxyadenosine 5'-monophosphate, conceivably may contribute to the adenine nucleotide degradation induced by deoxyadenosine in normal and malignant lymphocytes. PMID- 6290032 TI - Protection and potentiation of nitrogen mustard cytotoxicity by WR-2721. AB - The radioprotective agent WR-2721 was examined for its effects on modifying the cytotoxicity of HN2 against normal and tumor cells in the AKR mouse. Quantitation was carried out by the spleen colony assay for both normal hematopoietic stem cells and AKR leukemia cells. Protection from drug toxicity and normal cell cytotoxicity was noted. Potentiation of cytotoxicity to AKR leukemia was found. PMID- 6290033 TI - Differentiation of platelet-aggregating effects of human tumor cell lines based on inhibition studies with apyrase, hirudin, and phospholipase. PMID- 6290034 TI - Comparison of some carcinogenic, mutagenic, and biochemical properties of S vinylhomocysteine and ethionine. PMID- 6290036 TI - Epidermal growth factor receptors and effect of epidermal growth factor on growth of human breast cancer cells in long-term tissue culture. AB - Epidermal growth factor (EGF) may be important in regulating the proliferation of mammary epithelial cells. In the present study, we examined EGF binding and effect on growth in nine human mammary cell lines. The T-47D, MCF-7, SK-Br-3, AIAb 496, BT-20, and BT-474 tumor cell lines and a cell line (HBL-100) derived from milk exhibited EGF binding; both high (Ka 10(10) M-1)- and low (Ka 10(9) M 1)-affinity sites were detected. The total number of EGF receptors per cell of different cell lines varied from 1.6 X 10(3) sites/cell (for AIAb 496) to 1.5 X 10(6) sites/cell (for BT-20). The two floating cell lines, DU4475 and Lev III, had no detectable EGF binding. Effect of EGF on growth was studied by monitoring cell number and the incorporation of [3H]thymidine into DNA of cells maintained in Dulbecco's modified Eagle's medium supplemented with 0.1% fetal bovine serum. Using these procedures, only T-47D cells were stimulated by EGF at low concentrations (0.1 to 1 ng/ml). At concentrations higher than 10 ng/ml, EGF was inhibitory to varying degrees in most cell lines that contained EGF receptors. The growth of the two floating cell lines that had no detectable EGF binding was unaffected by EGF. Our results show that EGF receptors are not present in all human breast cancer cell lines. There is no apparent correlation between EGF binding and its mitogenic activity in cell lines with EGF receptors. EGF may have biological roles in human breast cancer other than growth regulation. PMID- 6290035 TI - Identification and characterization of a high-affinity saturable binding protein for the carcinogen benzo(a)pyrene. AB - A protein that binds the chemical carcinogen, benzo(a)pyrene (BP), in a high affinity and saturable manner has been identified in cell extracts from the AKR 2B mouse embryo cell line. The BP bound to the carcinogen-binding protein (CBP) was exchangeable in a time- and temperature-dependent fashion and has a Kd of 1.8 X 10(-9) M. Competitive binding studies indicate that chemical carcinogens [3 methylcholanthrene, benz(a)anthracene, dibenz(a,c)anthracene] and other inducers of aryl hydrocarbon hydroxylase (5,6- and 7,8-benzoflavone) compete to varying degrees with BP for binding. Steroids, 2,3,7,8-tetrachlorodibenzo-p-dioxin, and phenobarbital did not compete with BP for binding to the CBP, BP prevented the heat inactivation of CBP. Additional properties of CBP reveal it to have a Stokes' radius of 31 A, molecular weight of 61,000, frictional ratio of 1.2, an apparent isoelectric point of 5.2, and an S value of 4.8 in linear sucrose gradients. It was estimated that there are about 20,000 molecules of CBP per AKR 2B mouse embryo cell. The CBP was found in two nontransformed and one chemically transformed cell line; a fourth cell line A-431 (human vaginal carcinoma) had significantly reduced amounts of CBP. PMID- 6290037 TI - Efficient metabolism of benzo(a)pyrene at nanomolar concentrations by intact murine hepatoma cells. AB - We have studied the metabolism of benzo(a)pyrene (BP) by intact mouse hepatoma cells, at nM concentrations of the carcinogen, using an assay in which we directly measure the rate of BP fluorescence disappearance. The rate of BP metabolism is half-maximal, at limiting cell dilution, when the concentration of BP is about 4 nM. This apparent Km for BP metabolism is much lower than those reported previously for several reasons. (a) Partitioning of BP into cells markedly influences kinetic measurements, and we account for these effects. (b) Enzyme inducers can competitively inhibit BP metabolism and thus may introduce artifacts into kinetic measurements. (c) Under the conditions of this assay, phenolic BP metabolites are produced but do not accumulate, due to their further metabolism; therefore, assays of BP metabolism which measure the production of phenols, such as the commonly used aryl hydrocarbon hydroxylase assay, may markedly underestimate the rate of BP metabolism when intact cells and low substrate concentrations are used. Our results show that cells can efficiently metabolize BP when exposed to BP concentrations similar to those present in the environment. PMID- 6290038 TI - Effect of disulfiram (tetraethylthiuram disulfide) amd diethyldithiocarbamate on the bladder toxicity and antitumor activity of cyclophosphamide in mice. AB - Cyclophosphamide is the most commonly prescribed alkylating agent in clinical medicine. The usefulness of cyclophosphamide is often limited, however, by its propensity to cause hemorrhagic cystitis especially in children or patients receiving concomitant radiotherapy. Administration i.p. of cyclophosphamide at doses of 100 mg/kg or more to mice produced a significant increase in urinary bladder weight within 48 hr of treatment. The present studies demonstrate that disulfiram prevented cyclophosphamide-induced bladder damage when administered p.o. within 1 hr of cyclophosphamide treatment. Diethyldithiocarbamate, a sulfhydryl-containing metabolite of disulfiram, had identical uroprotective activity. Unlike disulfiram, diethyldithiocarbamate was effective only when administered 2 to 4 hr after cyclophosphamide. Disulfiram augmented slightly the antitumor activity of cyclophosphamide against L1210 murine leukemia in vivo when administered 30 min prior to cyclophosphamide. In contrast, diethyldithiocarbamate had no effect on the antitumor activity of cyclophosphamide when administered 4 hr after cyclophosphamide. PMID- 6290039 TI - Mutagenesis and DNA binding of benzo(a)pyrene in cocultures of rat hepatocytes and human fibroblasts. AB - The genotoxicity of benzo(a)pyrene (BP) was investigated in combined cultures of rat hepatocytes and human diploid fibroblasts. Freshly isolated rat hepatocytes were shown to activate BP to a species which bound to and damaged hepatocyte and fibroblast DNA. A significant increase in the hypoxanthine-guanine phosphoribosyltransferase mutation frequency was induced when 10 to 100 microM BP was added to the cocultures. A comparative analysis of the binding of BP metabolites to hepatocyte and fibroblast DNA revealed that approximately 4 times more [3H]BP metabolites were bound to the fibroblast DNA than were bound to the hepatocyte DNA (per microgram DNA). Activation of BP by the fibroblasts themselves was shown not to be the cause of the relatively greater binding of BP to fibroblast DNA than to the hepatocyte DNA. These results suggest that proximate and/or ultimately carcinogenic metabolites of BP are readily released from isolated hepatocytes and that the metabolites are sufficiently stable and long lived so as to bind to the DNA of an adjacent cell. The relative protection of the hepatocytic DNA from BP metabolites that generated in the cytoplasm of the hepatocyte may be significant in view of the observations that the liver is not under normal conditions a target of BP carcinogenicity in vivo. PMID- 6290040 TI - Synthesis of alpha 1-antichymotrypsin and alpha 1-acid glycoprotein by human breast epithelial cells. AB - Malignant and uninvolved human breast tissues were maintained in organ culture for 3 to 6 days. Under these conditions, the three-dimensional glandular architecture is maintained with the least disruption of tissue integrity. The biosynthesis and release of glycoproteins were studied by using the incorporation of [14C]glucosamine and [14C]leucine by the breast surgical specimens. Five major families of labeled glycoproteins were identified from culture supernatants using two-dimensional gel electrophoresis. Quantitative immunoprecipitation established that 16 to 30% of the total of labeled glycoproteins were recognized as normal serum components. Two of these glycoproteins were antigenically related to normal human serum components as demonstrated with crossed immunoelectrophoresis. Evidence was obtained for the active synthesis of alpha 1-antichymotrypsin and alpha 1-acid glycoprotein by human breast epithelial cells. alpha 1 Antichymotrypsin accounted for 0.9 to 7.8% of the biosynthetically labeled glycoproteins from organ culture supernatants. This component was 11.9% of the glycoproteins released by a monolayer culture of the established breast carcinoma cell line, MCF-7. alpha 1-Acid glycoprotein made up 0.7 to 3.1% of the labeled glycoproteins. alpha 1-Antichymotrypsin is a known neutral serine proteinase inhibitor with a particularly strong affinity for cathepsin G. alpha 1-Acid glycoprotein may function primarily as a potent immunomodulator by suppressing lymphoblastogenesis. These glycoproteins may thus have regulatory roles in the proteolytic modification of breast tissue and represent the tissue's own protecting shield against invading leukocytes. PMID- 6290041 TI - Effects of 12-O-tetradecanoylphorbol-13-acetate on the differentiation of simian virus 40-infected human keratinocytes. AB - We have studied the effects of the tumor promoter 12-O-tetradecanoylphorbol-13 acetate on the differentiation of human epidermal keratinocytes infected by the oncogenic virus Simian Virus 40. The cells in monolayers exposed to a concentration of 12-O-tetradecanoylphorbol-13-acetate as low as 5 X 10(-10) M redistributed into dense patches within 48 hr. This morphological change was accompanied by a 10- to 30-fold increase in the production of cells cytochemically stained by orange G:acid fuchsin indicating keratinization and a 2 to 3-fold increase in the exfoliation of cornified envelope-bearing cells. The induced cellular differentiation occurred in parallel with an inhibition of cell growth. The biologically inactive congener 4 alpha-phorbol-12,13-didecanoate at equimolar concentrations did not affect the growth or differentiation of these cells. Binding studies using [3H]12-O-tetradecanoylphorbol-13-acetate revealed a binding site with characteristics similar to those found for other cell types (Kd = 17 nM; 1.25 X 10(5) available sites/cell). PMID- 6290043 TI - Presence of a novel recombinant murine leukemia virus-like glycoprotein on the surface of virus-negative C57BL lymphoma cells. PMID- 6290042 TI - Human breast cell-mediated mutagenesis of mammalian cells by polycyclic aromatic hydrocarbons. AB - A system has been developed in which human breast cells activate chemical procarcinogens to mutagenic compounds. The degree of activation is quantitated by the estimation of induction of 6-thioguanine-resistant specific locus mutants in a cocultured Chinese hamster V-79 cell population which does not activate carcinogens. Both mammary stromal and parenchymal cells could activate the procarcinogen 7,12-dimethylbenz(a)anthracene. In addition, it is shown that the two mammary cell populations converted both 7,12-dimethylbenz(a)anthracene and benzo(a)pyrene to water-soluble metabolites. The stromal cells produced substantial amounts of glucuronic acid conjugates, but the parenchymal cells did not. Both cell types metabolize benzo(a)pyrene to the organic-soluble metabolites 9,10- and 7,8-dihydrodiol and both 9- and 3-hydroxybenzo(a)pyrene. These results suggest that the human breast may be a target for polycyclic aromatic hydrocarbon carcinogenesis. PMID- 6290044 TI - Effect of cell density on growth rate and amino acid transport in simian virus 40 transformed 3T3 cells. AB - Rate of proliferation and amino acid transport were assessed in simian virus 40 (SV40)-transformed 3T3 cells by measurements of growth rate quotient and L-serine uptake via System ASC, respectively. Growth rate and cell density of the cultures were varied by modifying: (a) the number of cells initially plated; and (b) the period spent by the cells in culture. The growth rate quotient of SV40 3T3 cells was not correlated with cell density. Sparse cultures exhibited marked fluctuations in their growth rate as a function of time, whereas, under comparable conditions, crowded cultures retained some form of growth control by density. Transport activity by System ASC decreased as a function of increased cell density following a complex trend described by a double-exponential equation. The density-dependent decrease in amino acid transport was not accompanied by a parallel change in the rate of cell proliferation. These results indicate that alterations in amino acid transport are not linked with cell growth and suggest that an increase in transport activity is not a prerequisite for an optimal rate of proliferation in SV40-transformed 3T3 cells. PMID- 6290045 TI - Distribution of myoepithelial cells and basement membrane proteins in the normal breast and in benign and malignant breast diseases. AB - An immunocytochemical method for fixed and paraffin-embedded human breast biopsies is reported for the detection of myoepithelial and epithelial cells using antibodies to myosin and keratin, respectively, and of basement membranes using antibodies to laminin and type IV collagen. Using these markers, myoepithelial cells can be clearly distinguished in the normal breast and in the benign breast diseases sclerosing adenosis, epitheliosis, and fibroadenoma. In sclerosing adenosis, myoepithelial cells form a major cellular component. A stromally derived spindle cell is identified which stains with myosin but not with keratin antibodies (myofibroblast). These cells are seen in one-fifth of the fibroadenomas. Although cells staining with myosin antibodies are seen in the infiltrating component of all 18 carcinomas examined, elongated cells staining with both myosin and keratin antibodies (myoepithelial-like) are seen in only one infiltrating carcinoma where they are interposed at the stromal-epithelial junction of the infiltrating tumor cells. In contrast to the situation in benign breast diseases, mature myoepithelial cells form a very minor component of the majority of infiltrating ductal carcinomas. Basement membrane proteins, laminin, and type IV collagen are present in normal breast, benign breast disease, and grade I infiltrating ductal carcinomas but are absent in carcinomas of grades II and III. PMID- 6290046 TI - Synergistic interaction of two classes of transforming growth factors from murine sarcoma cells. AB - Transforming growth factors (TGFs) isolated from murine sarcoma virus-transformed 3T3 cells have been separated by high-pressure liquid chromatography into two subsets. One subset, called TGF alpha, competes with epidermal growth factor (EGF) for receptor sites, whereas the other, called TGF beta, does not. TGB beta, purified by high-pressure liquid chromatography, will not induce formation of large colonies of cells in soft agar in the absence of TGF alpha or EGF. However, the combined action of either TGF alpha or EGF (which by themselves are relatively ineffective in promoting growth of cells in soft agar) together with TGF beta results in a potent synergistic effect, with formation of large colonies. Chemically modified analogs of EGF also potentiate TGF beta activity to the extent that they bind to the EGF receptor. It is suggested that TGF beta may be an important mediator of the known effects of both TGF alpha and EGF on neoplastic transformation. PMID- 6290047 TI - Metabolism and tumorigenicity of 7-, 8-, 9-, and 10-fluorobenzo(a)pyrenes. AB - The skin tumor-initiating activities of 7-, 8-, 9-, and 10-fluorobenzo(a)pyrenes have been compared to that of benzo(a)pyrene in female Sencar mice after 16 weeks of promotion with 12-O-tetradecanoylphorbol-13-acetate. Single initiating doses of 200 or 400 nmol of each hydrocarbon were tested, and the mice were treated twice weekly with 3.2 nmol of the promoter. Under these conditions, benzo(a)pyrene caused an average of 2.9 and 5.7 papillomas/mouse, respectively, whereas none of the four fluorinated hydrocarbons had significant tumor initiating activity. Examination of the hepatic metabolism of 7- and 8 fluorobenzo(a)pyrene revealed that a 7,8-dihydrodiol was not detected as a metabolite; thus, the bay-region diol-epoxide pathway known to be responsible for the tumorigenic activity of benzo(a)pyrene is blocked. Although 7,8-dihydrodiols are formed from 9- and 10-fluorobenzo(a)pyrene, these dihydrodiols with fluorine substituted on the 9,10-double bond may not be converted to diol-epoxides by the cytochrome P-450 system, or such fluorinated 7,8-diol-9,10-epoxides may not be tumorigenic. PMID- 6290048 TI - Effects of aminoglutethimide on delta 5-androstenediol metabolism in postmenopausal women with breast cancer. AB - delta-5-Androstene-3 beta, 17 beta-diol has potential estrogenic activity because it is known to bind to receptors and translocate to the nucleus of certain estrogen target tissues. Its role in the biology of breast cancer is unclear. Aminoglutethimide plus hydrocortisone ("medical adrenalectomy") has been used to treat postmenopausal women with metastatic breast cancer. We studied delta 5 androstene-3 beta, 17 beta-diol metabolism in postmenopausal women with breast cancer before and during aminoglutethimide-plus-hydrocortisone therapy, utilizing the constant infusion technique. The metabolic clearance rate for five subjects was 799 +/- 89 liters/24 hr (470 +/- 47 liters/24 hr/sq m) before and 751 +/- 93 liters/24 hr (444 +/- 57 liters/24 hr/sq m) during therapy. Plasma delta 5 androstene-3 beta, 17 beta-diol and delta 5-androstene-3 beta, 17 beta-diol free index decreased despite absence of change in the metabolic clearance rate. Increased dehydroepiandrosterone/delta 5-androstene-3 beta, 17 beta-diol conversion ratios in individual patients suggested an increase in 17 beta hydroxysteroid dehydrogenase activity during therapy. There were no alterations in the formation of the estrogen precursors testosterone and delta 4-androstene 3,17-dione. PMID- 6290049 TI - Doxorubicin-vincristine therapy for Wilms' tumor: a pilot study. AB - Doxorubicin plus vincristine chemotherapy was given to 31 children following nephrectomy for Wilms' tumor. Radiation therapy was used as indicated. Disease free survival by stage is: eight of nine patients (stage I), eight of nine (stage II), nine of ten (stage III), and two of three (stage IV). Median follow-up of survivors is 28 months (range, 2-67); for all but four patients, follow-up is greater than 12 months. Two of the three stage I-III failures occurred in children with unfavorable histologies; the third failure was due to fatal anthracycline cardiomyopathy. Lowering the maximal cumulative doxorubicin dose from 450 to 240 mg/m2 did not increase failures. Doxorubicin-vincristine appears to be effective chemotherapy for Wilms' tumor. PMID- 6290050 TI - Invited comment on the Milwaukee pilot study of doxorubicin-vincristine therapy for Wilms' tumor. PMID- 6290051 TI - Mitomycin, hexamethylmelamine, and vindesine as second-line treatment for small cell cancer of the lung. PMID- 6290052 TI - Intensive timed sequential combination chemotherapy in extensive-stage small cell carcinoma of the lung. PMID- 6290053 TI - Small-cell bronchogenic carcinoma--primary and relapse therapy with etoposide (VP 16), methotrexate and CCNU. PMID- 6290054 TI - Etoposide after ACO treatment for small-cell bronchogenic carcinoma. PMID- 6290055 TI - Etoposide single-agent chemotherapy for solid tumors. PMID- 6290056 TI - Etoposide salvage therapy for refractory germ cell tumors: an update. PMID- 6290057 TI - Etoposide as single agent and in combination chemotherapy of bronchogenic carcinoma. PMID- 6290058 TI - Combination cis-platinum and etoposide in small-cell lung cancer. PMID- 6290059 TI - Combination chemotherapy of small-cell bronchogenic carcinoma with etoposide (VP 16), ifosfamide and vindesine (VPIV), and with adriamycin, cis-platinum and vincristine (APO). PMID- 6290060 TI - Some new data on the distribution of benzo (a) pyrene in fresh water and seawater. AB - Data on the content of benzo(a)pyrene (BP) in various bodies of water in the Estonian SSR including lakes of agricultural regions, lakes and rivers in industrial areas, bodies of water in the National Park and nature protection areas used for drinking water supply, sea gulfs and bays used for recreation are reviewed. Some data on the distribution of BP in bottom sediments and biota of the aqueous medium as well as the accumulation of BP in various organs and tissues of different species of fish are reported. It has been found that the BP level in the bodies of water of the Estonian SSR is relatively low, the highest BP content has been found in seawater, but even there the average concentration of BP was below the safety limit for BP in bodies of water (0.005 microgram/l). Bottom sediments, algae, higher waterplants, zooplankton and fish accumulate BP where its content is about 1,000-100,000 times higher than in water. PMID- 6290062 TI - Steroid receptors in cystosarcoma phyllodes. AB - Seven cases of cystosarcoma phyllodes were studied. All but one of the patients were premenopausal. Histologic appearance permitted us to classify 5 as low-grade and 2 as high-grade tumors; 40% of the low-grade tumors were estrogen-receptor positive (ER-positive), 100% were progesterone receptor-positive (PR-positive), and 60% glucocorticoid receptor-positive (GR-positive). In these tumors, mean PR values were 26.4 fmoles/mg protein; mean GR values were 27.2 fmoles/mg protein; mean ER values were 10,0 fmoles/mg protein. ER and PR were present in one of the high-grade tumors at slightly higher values than in low-grade tumors. GR was present in both high-grade tumors at a mean concentration of 275 fmoles/mg protein. Sucrose gradient analyses showed only the presence of 4S-specific receptors for ER and PR. GR receptors sedimented faster (6-7S). PMID- 6290061 TI - Perchloric acid-soluble serum glycoprotein as an index for tumor cell burden in the early detection of cancer. AB - Neoplastic cells, particularly human mammary carcinoma cells, shed or secrete glycoproteins which are tumor-specific. These compounds contain N acetylneuraminic acid (NANA) and they differ from the bulk of serum proteins and glycoproteins in being soluble in perchloric acid. Graded number of R3230 adenocarcinoma (AdCa) cells were implanted subcutaneously in groups of female Fisher rats. At time intervals, while under anesthesia the spleens were dissected out, and blood was drawn from the animals. The blood was examined for NANA levels and the spleen cells for lymphocyte migration inhibition. The serum perchloric acid (PA) soluble proteins and the PA-NANA levels were time-dependent and increased with the number of implanted tumor cells. Maximum levels were found in sera from blood drawn 196 hours or later from animals which received 1,000 tumor cells/rat, or more. At 72 hours or more after tumor cell implantation, migration of splenic lymphocytes from animals which received 100 or more R3230 AdCa cells per animal was inhibited on contact with neuraminidase-treated formalinized R3230 AdCa cells. The magnitude of inhibition increased with time and with the number of implanted tumor cells. Therefore, blood PA-NANA levels and lymphocyte migration inhibition rae parameters for the in-vivo early detection and monitor tumor cell proliferation. PMID- 6290063 TI - Immunologic response in chickens from different genetic lines to Rous sarcoma virus. AB - Twelve young chickens of the Tumor Progressor and Tumor Regressor Lines were studied in respect to their humoral immunologic response to challenge with Rous sarcoma virus via inoculation into the wing web. On day 1, prior to virus inoculation and on days 10, 15, and 20 after injection of virus, the birds were bled and their serums tested for the presence of anti-RSV factors by standard procedures of serum-neutralization tests. From the conducting of statistical analyses of the data, it was determined that a strong association occurs between an early, increasing, and sustained presence of biologically active anti-RSV factors in the blood of the chicken and its ability to effect complete regression of the Rous sarcoma. PMID- 6290064 TI - Problems associated with the micropolarographic measurement of the arterial wall pO2. PMID- 6290065 TI - Research in neuromuscular disease. PMID- 6290066 TI - Clinical pharmacokinetics of cefotaxime and penetration in sputum, bone, and prostatic tissue. AB - Cefotaxime is a new powerful methoxycephalosporin with a broad anti-microbial spectrum, suitable for parenteral administration. In the present study, the concentrations of cefotaxime in serum and in bronchial secretion were determined after intramuscular injection of 1 gm every eight hours for seven days. Subjects were patients suffering from an exacerbation of chronic bronchitis. Serum levels versus time curve were interpreted in terms of a one-compartment open model. Pharmacokinetic parameters after single and multiple doses were investigated. No evidence of significant accumulation was found. Furthermore, a type of in vivo rate of killing with cefotaxime was investigated by evaluating the decrease in the number of colonies in bronchial mucus cultures daily for seven days. In two groups of volunteers who had to undergo surgery, bone and prostatic concentrations of cefotaxime were determined and correlated with serum levels. PMID- 6290067 TI - [Labile essential hypertension. The effect of Isoprenaline Spofa on plasma level of cyclic adenosine monophosphate]. PMID- 6290068 TI - [5 years of pyrophosphate scanning of the heart. A set of results from clinical practice]. PMID- 6290069 TI - Morphological studies on neuroglia. VI. Postnatal development of microglial cells. AB - The postnatal development of microglial cells was investigated in the neonatal rat brain by use of light- and electron microscopy, including enzyme histochemical techniques. Microglial cells were selectively stained by demonstration of their nucleoside diphosphatase (NDPase) activity and classified into three types: 1) In the early postnatal period "primitive microglial cells" showing scantily ramified processes were found in the cerebral cortex, the hippocampal formation, and the hypothalamus. During the course of the first postnatal week the processes of this cell type developed gradually and the cells were transformed into typical ramified microglial cells, called "resting microglial cells". 2) "Amoeboid microglial cells "showing typical features of macrophages were characteristic of the cerebral white matter. 3) "Round microglial cells" possessing a round soma and few pseudopodia but no characteristic processes occurred in large numbers in the subventricular zone of the lateral ventricle and as single elements in the vicinity of blood vessels. Histochemically, thiamine pyrophosphatase (TPPase) was demonstrated only in the fully developed, ramified microglial cells ("resting microglial cells"), which could be readily observed in the central nervous tissue from the age of 14 day. "Round and amoeboid microglial cells" did not show TPPase activity and disappeared after 14 days of postnatal life. By use of electron microscopy, in neonatal rats NDPase activity was apparent in the plasma membrane of the three types of microglial cells ("primitive, round, and amoeboid" types). They showed basically similar submicroscopic characteristics, i.e., well-developed Golgi apparatus, long strands of rough-surfaced endoplasmic reticulum, single dense bodies and vacuoles, and numerous ribosomes. "Amoeboid microglial cells" were characterized by their well-developed cytoplasmic vacuoles and phagocytic inclusion bodies. The present study strongly suggests a mesodermal origin for these microglial elements. PMID- 6290070 TI - The fine structure of the bovine Descemet's membrane with special reference to biochemical nature. AB - A freeze-etch replica method combined with biochemical analyses was used to investigate the ultrastructural organization of the bovine Descemet's membrane. The freeze-etch replica observations revealed that the intact Descemet's membranes were composed of stacks of two-dimensionally arranged hexagonal lattices, in which four components were resolved; (1) round densities as nodes, (2) rod-like structures connecting the densities, (3) randomly oriented fine filaments within the lattices, and (4) amorphous materials covering the lattices. When the membranes were treated with sodium dodecyl sulfate (SDS) and mercaptoethanol, only the amorphous materials were solubilized. However, both the amorphous materials and rod-like structures disappeared in SDS-mercaptoethanol urea-treated membranes. When the membranes were treated with a very low concentration (0.0005%) of collagenase, rod-like structures and round densities remained insoluble. If the concentration was raised to 0.01%, only the round densities persisted. Comparing these data with the amino acid analysis of each fraction, the following conclusions may be drawn: rod-like structures and fine filaments contain collagenous proteins of different solubility, while round densities and amorphous materials are non-collagenous in nature. PMID- 6290071 TI - Primary headaches: reduced circulating beta-lipotropin and beta-endorphin levels with impaired reactivity to acupuncture. AB - Eleven patients affected by common migraine (CM), eleven affected by daily chronic headache (DCH), and eight healthy volunteers were studied. Plasma levels of beta-endorphin (beta EP), beta-lipotropin (beta LPH). ACTH and cortisol were measured in basal conditions and after traditional Chinese acupuncture (TCA). Basal beta LPH and beta EP plasma levels (pg/ml) in the DCH patients (57.6 +/- 9.5 and 16.8 +/- 2.5, respectively; M +/- SE) were lower than those found in the controls (83.6 +/- 13.7 and 26.0 +/- 6.1; p less than 0.001), while those found in the CM cases showed inter-mediate values (75.3 +/- 12.0 and 24.4 +/- 5.8). ACTH and cortisol concentrations in both the CM and DCH patients were in the same range as those of the control group. TCA caused an increase in beta LPH and beta EP plasma concentrations in the control group (beta LPH: 117 +/- 16.9; beta EP: 44.1 +/- 6.7). Opioid plasma levels, however, remained unmodified after TCA in both the CM and DCH groups. ACTH plasma levels remained stable after TCA in all three subject groups. Patients suffering from primary headache are characterized by low beta LPH and beta EP plasma levels and by a poor reactivity of circulating opioids to non-stressful stimuli. PMID- 6290072 TI - Intermittent hypoendorphinaemia in migraine attack. AB - Beta-endorphin (RIA method, previous chromatographic extraction) was evaluated in plasma of migraine sufferers in free periods and during attacks. Decreased levels of the endogenous opioid peptide were found in plasma sampled during the attacks but not in free periods. Even chronic headache sufferers exhibited significantly lowered levels of beta-endorphin, when compared with control subjects with a negative personal and family history of head pains. The mechanism of the hypoendorphinaemia is unknown: lowered levels of the neuropeptide, which controls nociception, vegetative functions and hedonia, could be important in a syndrome such as migraine, characterized by pain, dysautonomia and anhedonia. The impairment of monoaminergic synapses ("empty neuron" condition) constantly present in sufferers from serious headaches, could be due to the fact that opioid neuropeptides, because of a receptoral or metabolic impairment, poorly modulate the respective monoaminergic neurons, resulting in imbalance of synaptic neurotransmission. PMID- 6290073 TI - Biosynthesis of polyprotein precursors to regulatory peptides. PMID- 6290074 TI - Absence of nucleosomes in a histone-containing nucleoprotein complex obtained by dissociation of purified SV40 virions. AB - Reduction of disulfide bonds involving the major capsid protein with dithiothreitol and removal of the calcium ions by EGTA disrupts the simian virus 40 virions. This process yields normal circular viral minichromosomes containing the four core histones and traces of the capsid proteins at pH values higher than 8.5. However, when carried out at pH 7.5, this procedure yields nucleoprotein complexes that contain both histones and the viral structural proteins. These pH 7.5 complexes appear as circular structures with a mean of 93 +/- 17 beads with a diameter of 7 nm and no visible nucleosomes when observed by electron microscopy. In contrast to the compaction of the viral DNA in minichromosomes, the length of these beaded structures is roughly the same as free DNA. We suggest that VP1, the major capsid protein, can act as a nucleosome unfolding agent in neutral pH and low ionic strength. PMID- 6290075 TI - Propagation of globin DNAase I-hypersensitive sites in absence of factors required for induction: a possible mechanism for determination. AB - We tested whether DNAase I-hypersensitive sites, once induced, can be propagated to daughter cells in the absence of the original inducer. Chicken embryo fibroblasts infected with a temperature-sensitive Rous sarcoma virus at 41 degrees C and then shifted to 36 degrees C become transformed and begin to transcribe globin RNA. DNAase I-hypersensitive sites appear in the alpha- and beta-globin chromatin domains. Neither transcription nor hypersensitive sites are detected in cells infected and maintained at 41 degrees C. Activation of the globin hypersensitive sites occurs within 30 min of a temperature shift to 36 degrees C and does not require new protein synthesis. To test for the self propagation of these hypersensitive structures, we inactivated the v-src gene product by a shift back up to 41 degrees C, and allowed the cells to divide 20 times at 41 degrees C. Although transcription of the globin genes was minimal after this treatment, the DNAase I-hypersensitive sites remained. The same sites can be induced by NaCl shock of cells. After the cells are returned to normal medium and allowed to grow for 20 doublings, the hypersensitive sites remain. This suggests that once formed, DNAase I-hypersensitive sites have the capacity to template their own structure independent of the initial "inductive" event. The single-stranded character of these DNAase I-hypersensitive sites could explain these results. PMID- 6290077 TI - Transposon-mediated site-specific recombination: identification of three binding sites for resolvase at the res sites of gamma delta and Tn3. AB - The tnpR gene product, resolvase, of the transposable element gamma delta mediates site-specific recombination between two copies of gamma delta directly repeated on the same replicon, and it negatively regulates transcription of the tnpA (transposase) gene and its own gene. The recombinational site, res, and the regulatory region both are located in the tnpA-tnpR intercistronic region. In studying the interaction of purified resolvase with DNA fragments derived from gamma delta and the related transposon, Tn3, that span this region, we have demonstrated that three sites specifically bind resolvase. Site I overlaps the recombinational crossover point and both transcriptional promoters. Sites II and III cover most of the DNA between the crossover point and the translational start of the tnpR gene. These are the only binding sites we have detected in a region of about 400 base pairs centered on the crossover point. Studies of cointegrates that contain only part of the region that binds resolvase indicate that site I is not sufficient for efficient site-specific recombination and suggest that all three sites are probably required. PMID- 6290076 TI - Differences in human alpha-, beta- and delta-globin gene expression in monkey kidney cells. AB - We have compared the function of the human alpha-, beta- and delta-globin genes using various plasmid expression vectors derived from pBR322. Amplification of recombinants occurred after their introduction, by calcium-phosphate-mediated DNA transfer, into monkey kidney cells that constitutively produce T antigen (COS cells). The human alpha-globin gene promoter functioned independently, but the beta-globin gene promoter was nearly totally dependent on the enhancing activity of the 72 bp direct repeats from the SV40 genome. Furthermore, when the human alpha- and beta-globin genes were linked in the same vector, the alpha promoter was active but the beta promoter was not. Function of the delta-globin gene promoter also depended on the enhancer element. In vectors containing the 72 bp repeats and the beta- or delta-globin gene, the activity of the beta-globin gene was approximately 50 times greater than that of the delta-globin gene, approximating the ratio of beta and delta mRNA observed in normal human bone marrow cells. PMID- 6290078 TI - Response of murine epidermis to 2,3,7,8-tetrachlorodibenzo-p-dioxin: interaction of the ah and hr loci. AB - 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) and related halogenated aromatic hydrocarbons produce epidermal hyperplasia, hyperkeratosis and sebaceous gland metaplasia in the skin of mice bearing the recessive mutation (hr/hr) hairless. This response is mediated through the cytosol receptor protein: the structure activity relationship for receptor binding corresponds to that for production of the skin lesion, and these histopathological changes segregate with the genetic polymorphism at the Ah locus, the locus determining the cytosol receptor. In HRS/J mice, an inbred strain segregating for the hr locus, both hairless (hr/hr) and haired (hr/+) mice possess the high-affinity cytosol receptor and respond to TCDD with the induction of epidermal aryl hydrocarbon hydroxylase activity, a receptor-mediated biochemical response; however, only hr/hr mice develop the proliferative/metaplastic skin response. We propose a genetic model for the interaction of the Ah and hr loci, to account for the differential response to TCDD observed in the skin of HRS/J hr/hr and hr/+ mice. PMID- 6290079 TI - A novel type of transposon generated by insertion element IS102 present in a pSC101 derivative. AB - We describe a novel type of transposon in the tetracycline resistance plasmid pYM103, a derivative of pSC101 carrying a single copy of an insertion element IS102. The new transposons we found were identified as DNA segments, approximately 6 kb (Tn1021) and 10 kb (Tn1022) in length, able to mediate the cointegration of pYM103 with plasmid Col E1. The resulting cointegrate contains either of these pYM103 segments duplicated in a direct orientation at the junctions of the parent plasmids. A direct duplication of a 9 bp sequence at the target site in Col E1 is found at the junctions for cointegration. Both transposons have IS102 at one end and also contain different lengths of the pYM103 DNA adjacent to IS102, including the tetracycline resistance gene. Each transposon contains terminal inverted repeats of a short nucleotide sequence. These results and the fact that IS102 can itself mediate plasmid cointegration, giving rise to a duplication of a 9 bp target sequence, indicate that IS102 is responsible for generation of Tn1021 and Tn1022. They are quite different from the common IS-associated transposons, which are always flanked by two copies of an IS element, and may be similar to transposons such as those of the Tn3 family and phage Mu. PMID- 6290080 TI - The herpes simplex virus amplicon: a new eucaryotic defective-virus cloning amplifying vector. AB - We have employed repeat units of herpes simplex virus (HSV) defective genomes to derive a cloning-amplifying vector (amplicon) that can replicate in eucaryotic cells in the presence of standard HSV helper virus. The design of the HSV amplicon system is based on the previous observation that cotransfection of cells with helper virus DNA and seed monomeric repeat units of HSV defective genomes results in the regeneration of concatemeric defective genomes composed of multiple reiterations of the seed repeats. Cotransfection of cells with helper virus DNA and chimeric repeat units containing bacterial plasmid pKC7 DNA resulted in the generation of defective genomes composed of reiterations of the seed HSV-pKC7 repeats. These chimeric defective genomes were packaged into virus particles and could be propagated in virus stocks, with the most enriched passages containing more than 90% chimeric defective genomes. Furthermore, monomeric chimeric repeat units could be transferred back and forth between bacteria and eucaryotic cells. A derivative vector constructed so as to contain several unique restriction enzyme sites could be potentially employed in the introduction of additional viral or eucaryotic DNA sequences into eucaryotic cells. PMID- 6290081 TI - An essential role for cyclic AMP in growth control: the case for yeast. PMID- 6290082 TI - A small tandem duplication is responsible for the unstable white-ivory mutation in Drosophila. AB - The white-ivory (wi) mutation, an unstable allele of the white locus in Drosophila, reverts to wild-type at frequencies of 5 X 10(-5) in homozygous females, and 5 X 10(-6) in males and deletion heterozygous females. We show by molecular cloning and Southern blot analysis of DNA from wi flies that a 2.9 kilobase tandem duplication within the white locus is responsible for the mutation. Phenotypic reversion appears, in most cases, to be due to an exact excision of the extra copy of the sequence. Two derivative alleles of wi, one phenotypically wild-type, the other a partial revertant, carry insertions of moderately repetitive DNA from outside the locus, in addition to suffering deletions of some white locus DNA. Earlier genetic data preclude unequal crossing over between homologs as an explanation for the precise reversions. Rather, an intrachromosomal meiotic event seems to be responsible. Our results suggest that intrachromosomal recombination may be responsible in other systems for a larger number of rearrangements than has been suspected, and that interallelic recombination frequencies in Drosophila do not always correlate in a simple way with DNA length or extent of homology. PMID- 6290084 TI - DNA gyrase is a host factor required for transposition of Tn5. AB - We show that DNA gyrase is required for transposition of Tn5. Coumermycin, a potent inhibitor of DNA gyrase subunit B, inhibits transposition in a wild-type strain, but has no effect on strains carrying a coumermycin-resistant allele in gyrB. In addition, strains containing a thermolabile subunit A of gyrase (gyrA43) are defective for transposition at a nonpermissive temperature. The requirement for gyrase is due to a requirement for supercoiled DNA. We showed this by introducing into the gyrA43 strain a deletion of the gene encoding topoisomerase I. The introduction of the second mutation caused an increase in the superhelical density of DNA as well as an increase in the transposition frequency. This also implies that if the DNA is supercoiled there is no further requirement for gyrase. Experiments with coumermycin support this, because the drug does not inhibit transposition if the recipient DNA remains supercoiled. This indicates that if the DNA acting as recipient of the transposon is deficient in supercoils, it will be a poor substrate for transposition. We also describe a system in which a gene on a multicopy plasmid can be efficiently introduced into the Escherichia coli chromosome. PMID- 6290085 TI - Regulation of the cell cycle in neuroblastoma cells: the role of ion transport. PMID- 6290083 TI - Structure of the Drosophila mutable allele, white-crimson, and its white-ivory and wild-type derivatives. AB - The white locus in Drosophila is required for a normal brick-red eye color; deletions of this locus result in a bleached-white eye color. The white-crimson (wc) allele of white was isolated as a partial revertant of another mutant white allele, white-ivory (wi), a mutation due to duplication of sequences within the white locus. The wc allele reverts at high frequencies to wild-type and wi phenotypes and generates white-eyed derivatives, including deletions with one endpoint at the white locus. We analyzed the structure of the wc allele by molecular cloning and by Southern blot analysis of genomic DNA and found that the wc phenotype results from the insertion of a 10 kilobase DNA sequence into the wi duplication. Five independent phenotypic revertants of wc to wi were examined, and in each case reversion was accompanied by apparently precise excision of the insertion. Reversion of wc to a wild-type phenotype in each of the six cases examined was mediated by excision of both the insertion and one copy of the wi duplication, restoring gene structure to wild-type. PMID- 6290086 TI - Intracellular antibody against Clostridium perfringens enterotoxin fails to counteract toxin-induced damage. PMID- 6290087 TI - Irreversible inhibition of water transport in erythrocytes by fluoresceinmercuric acetate. PMID- 6290088 TI - [Cerebral gliomas, their appearance in CT and rational therapy]. PMID- 6290089 TI - [On the perimyelographic appearance of glioblastoma multiforme diffusum]. PMID- 6290090 TI - Inhibitors of cyclic adenosine monophosphate phosphodiesterase in Polygala tenuifolia. PMID- 6290091 TI - Determination of SA-446 in human whole blood and urine by electron capture-gas liquid chromatography. PMID- 6290092 TI - [Immunoenzymatic assay of IgG and IgM to EBV and CMV in sera of patients with virus hepatitis and normal control]. PMID- 6290093 TI - [Congenital epulis of the newborn (granulous Abrikosov tumor). Remarks apropos of a personal case]. AB - The authors report a case of middle and superior gingival tumor in a newborn patient. Macroscopic and topographic aspects are a "congenital epulis". The surgical treatment is easy. Histologically finding shows a benign tumor with granulous cells called "Abrikossoff Tumor". The literature is reviewed to study histological aspect and pathogeny of this tumor. They try to distinguish the macroscopic aspect of the "congenital epulis" with the histologically finding. It is not systematically a granulous cells tumor. PMID- 6290095 TI - Enzyme histochemical characteristics of spontaneous and induced hepatocellular neoplasms in mice. AB - Hepatocellular neoplasms are known to differ in enzyme activity from the surrounding non-neoplastic liver. We have compared histochemically the enzyme activity of spontaneous hepatocellular tumors in mice with tumors induced by diethylnitrosamine and dieldrin. Some neoplasms had increased activity, others had decreased enzyme activity, yet other had the same activity as the surrounding liver. Alkaline phosphatase, glucose-6-phosphatase, succinic dehydrogenase and adenosine triphosphatase, as well as glycogen levels were studied. Carcinomas differed from adenomas in having elevated enzyme activity significantly more often than adenomas. However, the carcinomas showed elevated glycogen levels less frequently than adenomas. Histochemically, pulmonary metastases resembled the primary hepatocellular carcinomas from which they were derived. Tumors of dieldrin animals were notable in having increased activity of all the enzymes which we studied more frequently than tumors of diethylnitrosamine animals or of controls. Differences in enzyme activity between the three mouse strains were slight. PMID- 6290094 TI - Evaluation of weekly escalating doses of dichloromethotrexate in patients with hepatocellular carcinoma and other solid tumors. AB - Twenty-one patients with solid tumors were treated with weekly 6-h intravenous infusions of dichloromethotrexate (DCM), with escalating doses every other week. Frequently observed toxicities included leukopenia, thrombocytopenia, and mucositis. Nausea, vomiting, diarrhea, and elevation of hepatic enzymes and bilirubin occurred less often. The toxicity of DCM was dose-dependent; the maximum tolerated dosage excalation plan was 400 mg/m2 x 2 weeks, 800 mg/m2 x 2 weeks, and then 1,200 mg/m2 weekly. Plasma concentrations of DCM were measured during 61 infusions and apparent half-lives determined. The plasma elimination of DCM appears to be similar to that of methotrexate. Three objective tumor responses seen in the seven hepatocellular carcinoma patients treated warrant further investigation. PMID- 6290096 TI - Regulation of phosphorylase kinase in rat ventricular myocardium. Role of calmodulin. AB - Conversion of phosphorylase b to a which is catalyzed by the enzyme phosphorylase kinase is known to require Ca++. Trifluoperazine, an inhibitor of calmodulin dependent enzymes, was utilized in the present study to clarify the role in vivo of calcium-calmodulin regulation of phosphorylase kinase. Twenty-minute preperfusion of isolated rat ventricles with 10(-5) M trifluoperazine had no effect on basal levels of phosphorylase a but significantly attenuated phosphorylase activation induced by either calcium (3.75 mM) or isoproterenol (3 x 10(-9) M, 3 x 10(-8) M). The positive inotropic effect of both agents and cyclic adenosine 3',5'-monophosphate (cAMP) levels were not altered by trifluoperazine in the perfused hearts. In addition, no effects of 10(-5) M trifluoperazine were noted on beta-adrenergic receptor binding of [3H](+/ )carazolol or on adenylate cyclase activity. In vitro studies with partially purified rat cardiac phosphorylase kinase demonstrated 1.5- to 3-fold stimulation by exogenous calmodulin. The addition of 10(-5) M trifluoperazine prevented calmodulin stimulation but had little effect on activity in the absence of exogenous calmodulin. The present results suggest that reversible binding of calcium-calmodulin may represent a physiological means for regulating phosphorylase kinase activity in rat cardiac muscle. PMID- 6290098 TI - Uptake of iodine and radionuclides by the infarcted cardiac muscle. PMID- 6290097 TI - beta-Adrenergic receptor subtypes in the rat renal cortex. Selective regulation of beta 1-adrenergic receptors by pheochromocytoma. PMID- 6290099 TI - The late prognostic value of acute scintigraphic measurement of myocardial infarction size. AB - Infarct, perfusion and blood pool scintigraphy were performed in 62 patients during hospitalization for acute myocardial infarction. The largest measured infarct or perfusion image defect and left ventricular ejection fraction were related to the late prognosis determined a mean of 16 months after the event. Breakpoint values for all scintigraphic variables could separate those who were asymptomatic on follow-up from those who died. The best indicators for selection of survivors and nonsurvivors were a scintigraphic infarct size greater than or equal to 25 cm2 and a perfusion abnormality greater than or equal to 35% of the projected left ventricular area. Among patients with perfusion abnormalities above this limit, 61% died; 93% of those with small perfusion abnormalities survived. Scintigraphic measurements of relative myocardial perfusion and function best separated patients asymptomatic on follow-up from those who developed heart failure and also best identified those with an unfavorable evolution, who developed heart failure or died. Early scintigraphic parameters appeared more accurate than other clinical laboratory indicators for determining late prognosis and could be important in planning treatment after acute infarction. PMID- 6290101 TI - Cyclic GMP in the human erythrocyte. Intracellular levels and transport in normal subjects and chronic hemodialysis patients. AB - We have studied the intracellular levels of cyclic GMP in the red blood cells of chronic hemodialysis patients before and after dialysis treatment and in normal subjects. Cyclic GMP is present at levels (mean +/- 1SD) of 19.6 +/- 4.7 and 18.4 +/- 5.8 nM/L before and after treatment, respectively. The levels in normal subjects are 3.6 +/- 0.9. The uptake of cyclic GMP was also studied and the data demonstrate that the red blood cell is slightly permeable to cyclic GMP, suggesting that cyclic GMP uptake could substitute for guanylate cyclase activity in the human erythrocyte. PMID- 6290100 TI - Radionuclide angiographic assessment of global and segmental left ventricular function at rest and during exercise after coronary artery bypass graft surgery. AB - Left ventricular ejection fraction (LVEF) was measured by radionuclide angiography at rest and during supine bicycle exercise before and 3 months after coronary artery bypass graft surgery (CABG) in 20 patients with chronic stable angina. The right anterior oblique gated first-pass technique was used to assess LVEF response to maximal exercise (Wmax), while the left anterior oblique equilibrium-gated technique was used to assess LVEF and relative LV volume changes during graded submaximal exercise. Mean LVEF was unchanged at rest after CABG by both the first-pass (60 +/- 12% vs 60 +/- 12%) and equilibrium-gated (61 +/- 13% vs 62 +/- 13%) measurements. At Wmax, mean first-pass LVEF was significantly higher postoperatively than preoperatively (63 +/- 17% vs 53 +/- 17%; p less than 0.01) with a higher Wmax (750 +/- 182 vs 590 +/- 202 kpm/min; p less than 0.001) and higher rate-pressure product (302 +/- 59 vs 222 +/- 57 units; p less than 0.001). Similarly, equilibrium-gated LVEF levels during graded exercise, using stepwise regression analysis, were significantly higher postoperatively than preoperatively (p less than 0.001); at the highest graded work load, they averaged 63 +/- 19% postoperatively and 53 +/- 17% preoperatively, with higher work loads (500 +/- 190 vs 417 +/- 155; p less than 0.05) and higher rate-pressure products (271 +/- 55 vs 207 +/- 53; p less than 0.001). The increase in exercise LVEF after surgery was due to a marked decrease in the ratio, relative to resting values, of counts-based end-systolic volumes during submaximal exercise (preoperatively 1.91 +/- 1.04; postoperatively 1.14 +/ 0.46; p less than 0.01). The five subjects in whom LVEF decreased significantly during exercise postoperatively all had one or more blocked or stenosed grafts. This study documents, by two independent radionuclide techniques, an improved LVEF during exercise at an increased maximal work capacity and rate-pressure product 3 months after successful CABG. PMID- 6290102 TI - Evaluation of a modified alcohol dehydrogenase assay for the determination of ethanol in blood. AB - We evaluated a new alcohol dehydrogenase (EC 1.1.1.1) enzymic assay (ADH-glycine, Sigma Chemical Co.) for the determination of ethanol in blood. This assay differs from the manufacturer's previous assay (ADH-pyrophosphate) in that glycine replaces pyrophosphate as the buffer and hydrazine replaces semicarbazide as the trapping agent. The standard curve for the assay was linear over blood ethanol concentrations of 0.50-5.00 g/L. The reaction time of the assay was 10 min. At 1.00 g/L within-run and between-run CVs were 3.96% (n = 20) and 4.01% (n = 20), respectively. Mean analytical recovery of ethanol added to whole blood at 0.50 5.00 g/L was 99.7% (SD 2.6%). We performed 100 consecutive clinical and forensic determinations by the ADH-glycine assay, the ADH-pyrophosphate assay, and gas chromatography. Correlation coefficients of the results by least-square linear regression were 0.995 for ADH-pyrophosphate vs ADH-glycine, and 0.990 for gas chromatography vs ADH-glycine. The major advantage of the ADH-glycine assay over the ADH-pyrophosphate assay is the shorter reaction time, 10 min vs 30 min. PMID- 6290103 TI - An improved assay of mammalian collagenase activity, and its use to determine hepatic extracellular matrix susceptibility to degradation. AB - This rapid, sensitive method of determining collagenase (EC 3.4.24.7) activity incorporates several advantages of previous methods. Soluble [14C]acetylated collagen is prepared as the enzyme substrate and collagen-cleavage products are separated from noncleaved collagen by precipitation with dioxane/methanol. The assay is more reproducible than previous methods and has a lower detection limit, 15 mU of enzyme activity. We used the method in a competitive substrate assay with isolated extracellular hepatic matrix from cirrhotic and normal rat liver. Purified collagenase was consistently bound to normal rat matrix to a greater extent than to cirrhotic matrix, suggesting that in hepatic fibrosis the extracellular matrix is not as susceptible to collagenase degradation as that in normal liver. PMID- 6290104 TI - Radioimmunoassay of corticotropin from plasma. AB - We describe a specific and highly sensitive radioimmunoassay for corticotropin (ACTH) in human plasma. As little as 1.9 pg of circulating ACTH can be detected per milliliter (0.44 pmol/L). ACTH is first extracted from plasma samples by adsorption onto pretreated Vycor glass powder, eluted from the adsorbent by acetone, and then lyophilized. For purification of 125I-labeled ACTH, a small octadecasilyl silica column is used. The mean (and SD) concentration of ACTH in plasma from 18 ostensibly normal subjects was 10.3 (4.3) pmol/L. After overnight suppression with dexamethasone in seven of these normal subjects, their ACTH values fell to 2.62 (1.3) pmol/L (p less than 0.0005). This assay will permit clinical evaluation of ACTH plasma concentrations in clinical investigation and of the effects of drug administration on circulating ACTH. PMID- 6290105 TI - Quantitative determination of lecithin and sphingomyelin in amniotic fluid. AB - A quantitative method for lecithin and sphingomyelin is presented. In this method, which may be calibrated with a primary standard phosphate salt or pure phospholipids, 2 mL of amniotic fluid is extracted, and the solvent passed through a column of hydroxyapatite to remove all phospholipids except lecithin and sphingomyelin. After evaporation, the residue is treated with methanolic sodium hydroxide, which hydrolyzes lecithin to inorganic phosphate and a methyl ester. The unreacted sphingomyelin is removed by extraction, and the solvent evaporated. Both fractions are digested and assayed for phosphate by reduced phosphomolybdate. The total time required is 2 h. Analytical recoveries were 91 97%, and excellent agreement was found for controls. Amniotic fluid specimens were assayed to illustrate application of the proposed method. If this method is used for obtaining a lecithin/sphingomyelin ratio, a new ranges must be established, as this method is not equivalent to thin-layer chromatographic ones. PMID- 6290106 TI - Solid-phase colorimetric determination of potassium. AB - A nonpolar organic film (plasticized polyvinyl chloride) containing the ionophore valinomycin was incubated with an aqueous solution containing potassium ion and a detectable anion (erythrosin B). The amount of erythrosin B retained by the film after washing could be measured by absorbance or reflectance, and was directly related to the potassium concentration. This dye-binding method is quantitative for potassium and is suitable for both aqueous and serum-based solutions. There was no interference by sodium in the range found in serum. Several polyvinyl chloride plasticizers and anionic dyes and some other ionophores were found to be useful. The anion binding is thought to be restricted to the surface. PMID- 6290107 TI - Erythrocyte glycolysis in protein-energy malnutrition. AB - Erythrocyte glycolysis has been studied in the anaemia associated with protein energy malnutrition (PEM) in Kivu. Several results were compatible with a lowering of the mean age of the erythrocyte population, notably raised levels of glucose-6-phosphate, hexokinase, Na+-K+- adenosinetriphosphatases and potassium, and low sodium concentration. Non-significant differences were observed for glucose utilization, lactate formation, and for concentrations of fructose-6 phosphate, fructose-1,6-diphosphate, adenosine diphosphate and pyruvate kinase; there was no gross disturbance of cation transport. The level of adenosine triphosphate was slightly decreased and that of 2,3-bisphosphoglycerate was not elevated, in spite of anaemia. The latter could not be explained by an instability of this metabolite. It is concluded that slight erythrocyte glycolytic abnormalities may occur in the anaemia of Kivu PEM, but that they are not the main cause of the haemolysis observed in this syndrome. PMID- 6290108 TI - Semi-micro techniques for the genotyping of galactokinase and galactose-1 phosphate uridyltransferase. PMID- 6290109 TI - Recovery of hypothalamo-pituitary-adrenal function in asthmatics whose oral steroids have been stopped or reduced. AB - Hypothalamo-pituitary--adrenal (HPA) function was studied using tetracosactrin and insulin hypoglycemia tests in eleven asthmatic patients who were receiving or had taken oral steroids. The patients had been on prednisone, 5--20 mg/day for 2- 17 years. As the dose was reduced they used beclomethasone inhalation, up to 800 micrograms/day. Seven insulin hypoglycemia tests were performed when the patients had been off oral steroids for 15--37 months. Plasma cortisol responses were normal in all seven, three had subnormal responses of ACTH. In five patients serial tetracosactrin and insulin hypoglycaemia tests were performed during reduction of steroid dose. Two patterns of recovery of HPA function were observed. In one, both hypothalamo--pituitary and adrenal function seemed to return simultaneously, in the other normal response to tetracosactrin appeared before that to insulin hypoglycaemia. One patient had normal ACTH and cortisol responses to the stress of a vasovagal attack, but failed to respond to subsequent hypoglycaemia. We conclude that a normal response to tetracosactrin does not indicate recovery of the HPA axis after stopping prolonged oral steroid therapy for asthma, and that patients should continue to carry steroid therapy cards for at least 5 years after such treatment has been discontinued. PMID- 6290110 TI - Simultaneous study of somatotrophic and corticotrophic pituitary secretions during ornithine infusion test. AB - An ornithine infusion test was performed in fifty-four children with constitutional short stature. The ornithine infusion induced an elevated level of GH at 45 min (mean value = 873 pmol/l) and a similar rise of cortisol levels (mean value = 544 nmol/l). An important peak of ACTH appeared 15 min before the increase of cortisol. In three panhypopituitary dwarfs studied, no elevation of GH or cortisol was observed. The well tolerated ornithine infusion test allows the simultaneous study of the somatotropic and corticotropic pituitary secretions in children with delayed growth. PMID- 6290111 TI - [Experimental and clinical studies on somatomedin]. PMID- 6290112 TI - [Evaluation of determination of plasma cortisol concentration by enzyme immunoassay]. PMID- 6290113 TI - Metabolic defects in immunodeficiency diseases. AB - There are two ways to find the cause of primary immunodeficiency diseases. One approach is to start with the immune defect and work backwards, using in vitro techniques to define where the primary abnormality lies in the immune response. Immunologists favour this approach for obvious reasons; and it is not without virtue since, for example, it has shown that the defect in most cases of primary hypogammaglobulinaemia is due to a failure of B lymphocyte maturation. The alternative approach is to screen empirically for defects in biochemical pathways in the hope of finding a clue which will eventually lead to the underlying disorder. This is a sensible approach in diseases which are clearly inherited (e.g. X-linked hypogammaglobulinaemia) but is less attractive in a disease such as late onset hypogammaglobulinaemia which is not obviously inherited. In practice, such procedures involve screening the urine for abnormalities in the quality or quantity of excreted compounds. Another way is to screen for abnormalities in organelle integrity by measuring the activity of various enzymes in subcellular fractions. In reality, the clue to the metabolic defect is usually discovered by accident, the prime example in our field being the discovery of adenosine deaminase (ADA) deficiency. PMID- 6290114 TI - Humoral immune response in Epstein-Barr virus infections. I. Elevated serum concentration of the IgG1 subclass in infectious mononucleosis and nasopharyngeal carcinoma. AB - Using radial immunodiffusion we measured IgG subclass concentrations and studied their distribution in serum samples from patients with infectious mononucleosis (IM) and nasopharyngeal carcinoma (NPC), two Epstein-Barr virus (EBV)-associated diseases, in comparison with two control groups [completely anti-EBV negative persons and subjects carrying antibodies to the viral capsid antigen (VCA)]. Antibody titres to VCA and to the early antigen (EA) were determined by indirect immunofluorescence and revealed characteristic patterns for the respective diagnostic groups. Nephelometric assays served for quantitating total protein, albumin, total IgG, IgA and IgM in all the sera. In the IM and NPC groups the concentration of IgG1 was significantly elevated by more than 50% whereas the other three subclasses remained unchanged as compared with the controls. Correspondingly, we found a significant increase of total IgG in IM and NPC. In IM, the only disease where VCA-specific IgM antibodies have been reported to occur, IgM levels were markedly elevated. Our data suggest that the IgG1 subclass plays an important role in the humoral immune response to EBV-determined antigens and that it is possibly involved in the control of virus infection. PMID- 6290115 TI - Effects of muramyl dipeptide on superoxide anion release and on anti-microbial activity of human macrophages. AB - Exposure of human monocyte derived macrophages to muramyl dipeptide in vitro increased phorbol myristate acetate-stimulated O2- release by these cells; the activity of such macrophages against Toxoplasma gondii or against Staphylococcus aureus was not increased. The failure of muramyl dipeptide to enhance human macrophage anti-microbial activity correlated with the failure of muramyl dipeptide to enhance O2- release in response to phagocytic stimuli. These results and those previously published by others indicate that muramyl dipeptide may enhance release of certain inflammatory mediators (O2- and endogenous pyrogen) without enhancing the anti-microbial activity of human macrophages. PMID- 6290116 TI - Defects of serum chemoattraction and polymorphonuclear leucocyte movement in patients with primary hepatocellular carcinoma. AB - Movement of polymorphonuclear leucocytes to the site of tumour cells may be an important stage in host defences against tumours in a variety of organs. In this study, sera from 29 of 30 patients with primary hepatocellular carcinoma had reduced ability to stimulate the movement in vitro of normal polymorphonuclear leucocytes. The serum defect was more severe in 11 patients with underlying cirrhosis but was not related to abnormalities of tests of liver function, levels of serum alphafetoprotein, or deficiency of complement factors C3 and C5. Serial studies showed that the defect was persistent and progressive in patients in whom the tumour did not respond to treatment. In 35% of patients, mainly those with cirrhosis, the sera contained antagonists to normal serum chemotactic factors which were heat stable and dialysable, but could be distinguished by their effect on complement factor C5a. A heat labile dialysable antagonist(s) was found in sera from 28% of the patients (mainly those without cirrhosis) which antagonized the movement of normal polymorphonuclear leucocytes (cell directed antagonism). In addition to these serum defects, polymorphonuclear leucocytes from two of seven patients studied had reduced movement which was not related to the presence in the serum of cell directed antagonists. These serum and cellular defects have not been reported previously in patients with primary hepatocellular carcinoma, and could compromise the body's defences against the tumour. PMID- 6290117 TI - The effects of imidazole on contractility and cyclic AMP levels of guinea-pig tracheal smooth muscle. AB - 1. Imidazole contracted guinea-pig isolated tracheal smooth muscle, and at low concentrations (50 mumol/l to 2 mmol/l) also enhanced contractions induced by histamine. 2. Imidazole had no effect on contractile responses to potassium chloride, acetylcholine, prostaglandin F2 alpha, or the ionophore A23187; nor did it affect anaphylactic contractions in tracheal strips from guniea-pigs sensitized to ovalbumin. 3. Imidazole had no effect on the content on cyclic AMP in guinea-pig tracheal smooth muscle, nor did it affect the accumulation of cyclic AMP induced by histamine, isoprenaline, theophylline plus isoprenaline, or prostaglandin E1. 4. Imidazole-induced contractions were partially antagonized by atropine, mepyramine maleate and the omission of extracellular calcium ions. 5. Imidazole-induced hyperreactivity to histamine was not affected by atropine or indomethacin, and was more pronounced in the absence of extracellular calcium ions. 6. It is concluded that imidazole does not affect cyclic nucleotide phosphodiesterase activity in guinea-pig trachea, and its effects on contractile responses of the smooth muscle are unrelated to any action of cyclic AMP metabolism. The potentiating effect of imidazole may involve inhibition of histaminase, or the facilitation of calcium ion release from intracellular storage sites. PMID- 6290119 TI - Methylaplysinopsin: a natural product of marine origin with effects on serotonergic neurotransmission. AB - 1. Methylaplysinopsin is an indolyl-methylene derivative of creatinin which was isolated from a sponge collected on the Great Barrier Reef. 2. It is a potent inhibitor of tetrabenazine-induced responses in rats and mice and appears to act through several mechanisms, each of which enhances central serotonergic function. 3. As a reversible inhibitor of type A monoamine oxidase (MAO) methylaplysinopsin is relatively short-acting and has a Ki of 0.2 mumol/l when serotonin is the substrate used. 4. Methylaplysinopsin (10(-4) mol/l) also releases [3H]-serotonin from prelabelled synaptosomes, but has little effect on release of [3H] noradrenaline ([3H]-NA) from a similar preparation. 5. In blocking uptake of [3H] serotonin into cerebral cortical synaptosomes, methylaplysinopsin has a potency similar to that of imipramine (Ki = 2 X 10(-7) mol/l cf. 5 X 10(-7) mol/l), but it is considerably less potent in blocking [3H]-NA uptake. 6. Methylaplysinopsin is a weak displacer of [3H]-serotonin specific binding (IC50 = 160 mumol/l for hippocampal membrane preparations and 66 mumol/l for crude homogenates of rat brain). 7. Thus, methylaplysinopsin has effects on various aspects of serotonergic function. Whilst some of these effects are weak, they appear relatively selective for this neurochemical system. PMID- 6290118 TI - The affinity of 17 alpha-hydroxyprogesterone and 17 alpha, 20 alpha dihydroxyprogesterone for classical mineralocorticoid or glucocorticoid receptors. PMID- 6290121 TI - Maternal cytomegalovirus infection and perinatal transmission. PMID- 6290120 TI - Effect of complement depletion on hepatic amoebiasis in hamsters. PMID- 6290122 TI - Prevention of viral infections during pregnancy. PMID- 6290123 TI - [Progressive supranuclear palsy with unusual argentophilic inclusion bodies in neurons]. PMID- 6290124 TI - [Adrenoleukomyeloneuropathy presenting as hyperpigmentation, neuropathy, visual disturbance and hearing loss]. PMID- 6290125 TI - [A case of chronically progressed peripheral neuropathy, spastic paraplegia, cerebellar ataxia and dementia associated with decreased alpha-galactosidase activity]. PMID- 6290126 TI - [Polyneuropathy accompanied by restrictive cardiomyopathy in primary systemic amyloidosis. A case report]. PMID- 6290127 TI - [A case of palpable nodular type of muscular sarcoidosis with special reference to serum angiotensin-converting enzyme (SACE)]. PMID- 6290128 TI - Should inferior venacavography be performed in management of children with Wilms' tumor? PMID- 6290129 TI - Reversal of human lymphocyte beta-adrenoceptor desensitization by glucocorticoids. AB - To investigate the effect of glucocorticoids on beta-agonist-induced desensitization, we studied the effect of a single intravenous dose of methylprednisolone (2 mg/kg) on beta-receptor density and affinity in lymphocytes from four normal and four mildly asthmatic subjects at the end of 3 to 5 wk of terbutaline therapy and from four normal subjects taking no other drug. Terbutaline decreased (-)[3H]-dihydroalprenolol binding sites by 53% in normal and by 42% in asthmatic subjects. Methylprednisolone restored the number of binding sites to levels statistically indistinguishable from the preterbutaline values in both groups of subjects. In subjects not exposed to terbutaline beforehand there was no significant alteration in receptor density after methylprednisolone, nor in normal lymphocytes incubated in vitro for 90 min with hydrocortisone (10(-5)M). No significant change in the dissociation constant was observed in any situation. A single intravenous dose of methylprednisolone reverses terbutaline-induced down-regulation of beta-adrenoceptors. This may provide a mechanism for the beneficial effect of steroids in restoring catecholamine responsiveness in asthmatic subjects. PMID- 6290130 TI - Temporal enhancement of renin-aldosterone blockade by enalapril, an angiotensin converting enzyme inhibitor. AB - Interruption of the renin-aldosterone system with angiotensin-converting enzyme inhibitors (CEI) should result in a low aldosterone secretion, but most investigators have measured aldosterone production only indirectly by plasma aldosterone (PA) levels or urinary metabolites. We evaluated the effects of CEI of the aldosterone secretion rate (ASR) and compared them with PA, urinary tetrahydroaldosterone (THA), plasma renin activity (PRA), and electrolyte balance in six normotensive subjects in a metabolic unit during a control period (5 days) and during administration of 10 mg/day enalapril for 28 days. Our results demonstrated that (1) the ASR did not decline until after 1 wk of CEI therapy and this was reflected by a corresponding decline in the urine potassium:sodium ratio, (2) upright PA levels at day 1 declined, but supine PA levels were unchanged, (3) THA excretion remained essentially unchanged and the THA:ASR ratio rose progressively during therapy, (4) PRA rose and was maximal on day 3, but subsequently declined. In conclusion, enalapril-induced hypoaldosteronism required several days to become demonstrable and this was not accurately assessed by PA or THA--possibly due, in part, to altered aldosterone metabolism. The simultaneous decline in both PRA and ASR could be due to a decrease in renin substrate. Caution is therefore warranted when assessing aldosterone secretion indirectly by either PA levels or urinary metabolites during CEI therapy. PMID- 6290131 TI - Radionuclide assessment of right ventricular ejection fraction: a comparison of first pass studies with 133Xe and 99Tcm. AB - Right ventricular ejection fraction (RVEF) was determine in 26 patients using the first pass radionuclide angiocardiography with 133Xe and 99Tcm as tracers. A good correlation (r = 0.88) was found. Duplicate determinations in 13 patients with 133Xe showed good reproducibility, the absolute value for the standard error of a single determination being 2.6% and the coefficient of variation 5.7%. In a reference group of 17 individuals RVEF was 49 +/- 5% (range 42-61%). It is concluded that 133Xe can be used for RVEF determinations. Due to the rapid elimination and the low radiation dose 133Xe is preferable when repeated determinations of RVEF are desired. PMID- 6290132 TI - Influence of fibre ingestion on carbohydrate utilization and absorption. AB - To evaluate the mechanism whereby the addition of fibre to the diet lowers postprandial plasma glucose levels, the effects of ingestion of 10 g guar gum, a non-absorbable storage polysaccharide, on carbohydrate utilization and absorption were examined in healthy subjects. Total glucose utilization independent of gastrointestinal absorption of glucose was determined by the hyperglycaemic clamp technique in which constant hyperglycaemia is maintained with a variable intravenous infusion of glucose. Net splanchnic glucose uptake was also measured. As compare to a control study without guar, ingestion of guar failed to increase total glucose utilization or the uptake of glucose by splanchnic tissues. In contrast, when guar was ingested together with 25 g D-xylose, a non-metabolizable pentose, blood xylose levels were 25-40% lower than when xylose was ingested alone (P less than 0.01). We conclude that the plasma-glucose-lowering effects of high-fibre diets are a consequence of decreased carbohydrate absorption rather than increased total glucose utilization or augmented uptake of glucose by the liver. PMID- 6290133 TI - Viral diseases: infections of the gastrointestinal tract. AB - In the 1970s investigations generated a remarkable amount of information on the viruses responsible for acute gastroenteritis. The two viruses responsible for most cases occur in epidemiologically distinct clinical forms. Although occasionally epidemic, rotavirus infections are usually sporadic and primarily affect infants and young children. This illness is characterized by severe diarrhea commonly lasting five to eight days and frequently associated with upper respiratory tract symptoms, fever, and vomiting. The Norwalk agent and Norwalk like viruses are characteristically epidemic and responsible for community-wide outbreaks of gastroenteritis among all ages. The disease this agent produces usually lasts 24 to 48 hours and is characterized by various combinations of vomiting, diarrhea, nausea, headache, and low-grade fever. PMID- 6290134 TI - Extraction of latent-type collagenase from cultured bovine dental pulps with NaCl or urea or by collagen degradation. AB - All of the collagenase activity extracted from cultured bovine dental pulp tissue with NaCl or urea solutions was due to enzyme in a latent form and identified as a typical animal collagenase. Isotonic sucrose solution solubilized no detectable collagenase activity from cultured dental pulps. Also, no collagenase activity was extracted from either fresh bovine dental pulps or those cultured in Tyrode's solution containing 50 micrograms/ml cycloheximide. A two-day difference was observed between the appearance of collagenase activity in the cultured pulps and in the culture medium. The activity profiles in the culture media showed essentially no difference with or without the addition of cyclohexamide on and after the 10th day of culture, indicating that the biosynthesis of collagenase in the cultured pulp might have terminated around that time. About 80% of the total pulp collagenase activity was extracted by a bacterial collagenase procedure. Nearly half (43.5%) of the collagenase activity extracted from the cultured pulps with NaCl or urea solution was precipitated with collagen molecules in the presence NaCl, and most of the precipitated activity retained in the precipitate even after washing it with NaCl-buffer solution. These facts suggest a close association of collagenase with the collagen in cultured dental pulp tissue. PMID- 6290135 TI - Cellular response to rabies virus infection. AB - The effects of rabies virus on host cells were studied and compared to those obtained with another rhabdovirus, vesicular stomatitis virus [J. Virol. 34, 777 781 (1980)]. We show here: (1) that rabies infection has no effect on cell morphology, while infection with vesicular stomatitis virus caused cell retraction. Thus, only vesicular stomatitis virus induced a depolymerization of the microfilaments; and (2) that microtubules and microfilaments do not play a major role in rabies virus production, as it is suggested by results obtained with several effectors (colcemid, colchicine and cytochalasin-B) which directly or indirectly affect cytoskeleton organization. The same properties were observed with directly or indirectly affect cytoskeleton organization. The same properties were observed with vesicular stomatitis virus. Furthermore, the use of cytochalasin-B shows that an inhibition of glycosylation of the virion spike protein occurs only in rabies infected cells. As vesicular stomatitis viral glycosylation is normal in cytochalasin-B treated cells, results obtained indicate that two types of interactions can occur between a virion and the host cell depending on the rhabdovirus type. PMID- 6290136 TI - Phylogeny and ontogeny of the phosphoglycerate mutases.--V. Inactivation of phosphoglycerate mutase isozymes by histidine-specific reagents. AB - 1. The three isozymes of glycerate-2,3-P2 dependent phosphoglycerate mutase present in tissues of mammals and reptiles were inactivated by both treatment with diethylpyrocarbonate and photooxidation with rose bengal. 2. Inactivation of type M isozyme purified from rabbit muscle was complete when two histidine residues per enzyme subunit were carboethoxylated. Hydroxylamine removed the carboethoxy groups, with partial recovery of the enzymatic activity. The cofactor protected the enzyme against inactivation. 3. The inactivation of rabbit muscle phosphoglycerate mutase by photooxidation with methylene blue and rose bengal was sharply pH dependent. The pH profile of enzyme inactivation followed the titration curve of histidine, suggesting that this amino acid was critical for enzyme activity. Glycerate-2,3-P2 did not protect phosphoglycerate mutase against photoinactivation. PMID- 6290138 TI - Hormonal effects on the liver glucose metabolism in rainbow trout (Salmo gairdneri). AB - 1. Glucagon, adrenaline and dibutyril cyclic AMP increased the release of glucose to the medium during incubation of liver slices from rainbow trout (Salmo gairdneri) while insulin had no effect. 2. Glycogen content decreased only slightly after cyclic AMP addition and even increased in the presence of glucagon and adrenaline. Consequently, the release of glucose was due mainly to gluconeogenesis. 3. This is corroborated by the reduction of glucose liberation in presence of alpha-cyanocinnamate, an inhibitor of gluconeogenesis. PMID- 6290137 TI - The isoelectric points of cytochromes in the reduced and oxidised forms. PMID- 6290139 TI - Heterogeneity in the liver mitochondria of the tadpole, Rana catesbeiana. AB - 1. The heterogeneity of liver mitochondria of the tadpoles, Rana catesbeiana, undergoing metamorphosis was investigated by a combination of pulse-chase labeling of mitochondria with [methyl-3H]thymidine and centrifugation of mitochondria on a density gradient of metrizamide. 2. The liver mitochondria of tadpole at premetamorphic stage are separated into two populations with mean densities of 1.128 (M2) and 1.112 (M3). 3. At metamorphic stage a population with mean densities of 1.174 (M1) appears additionally. 4. The activity of mitochondria to take up [methyl-3H]thymidine in vivo is 2-3 times higher at metamorphic stage than at premetamorphic stage. 5. The M1 population has a prominently high activity to take up L-[4.5-3H]leucine in vitro and also a high specific activity of cytochrome c oxidase. 6. These findings suggest that the mitochondrial populations found are of alternate stages in the mitochondrial maturation. PMID- 6290140 TI - Water content of equine articular cartilage: effects of enzymatic degradation and "artificial fibrillation". AB - A method is described for accurate and reproducible determination of wet and dry weights of small articular cartilage samples. The water content of the articular cartilage samples before and after treatment with selective degradative enzymes and "artificial fibrillation" was determined. It was found that the water content of articular cartilage can be altered by changing the physical and chemical composition of articular cartilage. The combination of protein polysaccharide degradation and "fibrillation" causes an increase in articular cartilage water content, while collagenase and the combination of collagenase and "fibrillation" causes a decrease. The relationship between these findings and function of normal and osteoarthritic cartilage is discussed. PMID- 6290141 TI - Computed tomography of the gastrointestinal tract. PMID- 6290142 TI - Bronchoalveolar lavage, serum angiotensin-converting enzyme, and gallium-67 scanning in extrathoracic sarcoidosis. AB - Results of bronchoalveolar lavage (BAL), 67Ga scanning, and serum angiotensin converting enzyme (SACE) assay are compared in the assessment of pulmonary involvement in ten cases of extrathoracic sarcoidosis. Standard clinical, radiologic, and pulmonary function tests detected no pulmonary changes in these patients, but BAL demonstrated an increased alveolar lymphocytosis in eight of ten cases. SACE levels were increased in two cases, and the thoracic gallium uptake was normal in all cases. BAL appears to be the best technique for diagnosing latent pulmonary involvement in extrathoracic sarcoidosis. PMID- 6290143 TI - Longitudinal observations of serum angiotensin-converting enzyme activity in sarcoidosis with and without treatment. AB - Serial measurements of serum angiotensin-converting enzyme activity were made to estimate its usefulness in following the course of 17 patients with sarcoidosis. In nine cases with spontaneous remissions, the enzyme levels decreased gradually, accompanied by improvements in chest radiologic findings. In eight patients given corticosteroids, the enzyme levels decreased rapidly preceding improvements in the chest roentgenograms. The levels returned to pretreatment values when there was radiologic relapse of disease. Reelevation of the enzyme level was also observed without determination of the chest radiologic findings in four of five patients who responded to therapy, but the elevated enzyme level remained lower than the pretreatment level. These observations suggest that the serum angiotensin-converting enzyme level reflects the activity of disease in untreated and corticosteroid-treated patients with sarcoidosis. However, a partial reelevation of the decreased enzyme activity in corticosteroid-treated patients does not necessarily indicate a relapse of the disease. PMID- 6290145 TI - Pancoast's syndrome and small cell lung cancer. AB - Four patients with small cell lung cancer (SCC) presenting with Pancoast's syndrome are described. Superior sulcus tumors are usually caused by epidermoid carcinoma or adenocarcinoma of the lung, and are routinely treated with radiotherapy followed by radical surgery. SCC, on the other hand, is widely disseminated at diagnosis and is best treated with chemotherapy. Although not previously reported as a cause of Pancoast's tumor, these four cases of SCC presenting as such clearly indicate the need for pretreatment histologic diagnosis to avoid unnecessary surgical intervention. Transcutaneous needle aspiration biopsy is a means by which the diagnosis can be safely made in patients presenting with apical lung tumors. PMID- 6290144 TI - Sympathoadrenal reactivity in exercise-induced asthma. AB - The possibility that sympathoadrenal activity is altered in asthma was examined in eight patients with a history of exercise-induced asthma (EIA), eight matched patients with nonexercise induced asthma (NEIA), and eight matched healthy control subjects. No medication was allowed for at least one week before examination. In a pretrial exercise test diagnosis of EIA was confirmed and each individual's work capacity (Vo2 max) was determined. The trial consisted of an orthostatic test and a standardized exercise test at 80 to 90 percent of VO2 max on a treadmill. The trial exercise test caused a decrease in FEV1 in EIA patients only, whereas measurements of Sgaw revealed a significant but less pronounced postexercise bronchoconstriction in NEIA-patients as well. Basal plasma catecholamine levels were similar in all groups. Noradrenaline and adrenaline levels were approximately doubled by the orthostatic test and increased approximately ten-fold following exercise, with no differences between the groups. Plasma cAMP levels were approximately doubled by the exercise test. In the EIA patients there was an inverse correlation between increases in plasma cAMP and decreases in Sgaw. Our study does not support earlier claims that exaggerated catecholamine response to exercise causes postexercise bronchoconstriction by alpha-adrenoceptor stimulation in EIA. Differences in study results appear to have methodologic explanations. PMID- 6290146 TI - An improved method of right ventricular gated equilibrium blood pool radionuclide ventriculography. AB - Gated blood pool radionuclide ventriculography provides a means for obtaining repeated studies of both cardiac ventricles with a single dose of radionuclide. Quantitative assessment of right ventricular (RV) function using this technique has been complicated by several technical problems. We describe a new method of RV blood pool analysis which attempts to solve these problems using well established concepts for left ventricular (LV) blood pool analysis: (1) variable regions of interest; (2) computer edge detection with operator intervention; and (3) computer selected background. Results showed a strong linear correlation between gated first pass RV ejection fraction (RVEF) and the gated blood pool RVEF (n = 22; r = 0.93; blood pool RVEF = 0.03 + 0.89 X first pass RVEF; Sy.x = 0.04). There was also a strong linear correlation between LV and RV stroke counts in patients without valvular regurgitation, intracardiac shunts, or ventricular aneurysms (n = 19; r =0.86; RV counts = 72 + 0.94 X LV counts; Sy.x = 116). In terms of both of these validation standards this method proved superior to three published methods of RV blood pool analysis that used hand-drawn regions, and is suitable for analysis of rest, exercise, and intervention studies of RV function. PMID- 6290147 TI - Melanotic neuroectodermal tumor of the neurocranium in infancy. AB - Melanotic neuroectodermal tumors of the neurocranium are a rare but life threatening disorder of infancy. 11 previously reported cases are reviewed in terms of clinical presentation, radiological diagnosis, and management. A twelfth case, a 4-month-old infant who developed three discrete sites of tumor unilaterally in the neurocranium is presented. Several hypotheses for the mechanism of formation of these tumors are reviewed. The authors propose that the mechanism of formation involves a dysontogenesis of neural crest tissue and that these tumors form, at least in part, from fragments of melanin-containing arachnoid villi which are displaced during embryonic development. PMID- 6290149 TI - Extraction and serologic study of hepatitis A antigen. PMID- 6290148 TI - CT and ACTH treatment in infantile spasms. AB - Computed tomography of 8 cases with West's syndrome before, during and after ACTH treatment are reported. The scans, performed at the third week of therapy, showed consistent widening of the sulci, cisterns and ventricles in all the patients. Of these, 2 patients underwent ICP monitoring which showed higher than normal values. A return to the normal ICP values in association with the disappearance of the CT findings was observed in both cases. It is concluded that widening of the sulci, cisterns and ventricles are not findings of atrophy, but a condition of initial communicating hydrocephalus, which is in accordance with the hypotheses of Riikonen and Lyen. PMID- 6290150 TI - Cytomegalovirus infection of the uterine cervix. Report of 2 cases. PMID- 6290152 TI - [Diagnostic problems of insulinoma--with analysis of clinical diagnosis of 61 cases]. PMID- 6290151 TI - Polymorphism and stability in the histone gene cluster of Drosophila melanogaster. AB - Histone genes in Drosophila melanogaster are organized into repeats of 4.8 and 5.0 kb (Lifton et al., 1978). We find these repeat sizes in every one of the more than 20 Drosophila strains we have examined. Strains differ in the relative amounts of the two repeat types, with ratios varying from 1:1 to 1:4, the 5.0 kb repeat always present in equal to or greater amounts than the 4.8 kb repeat. Restriction enzyme digestion and blotting analysis reveals that the strains also differ in a number of far less abundant fragments containing histone DNA sequences. In the Amherst and Samarkand strains, there are, in addition, many copies of 4.0 and 5.5 kb repeat-like fragments respectively. A series of stocks were made isogenic for single second chromosomes from the Amherst strain. The hybridization patterns of the histone DNA from these stocks containing different Amherst chromosomes are very similar but a number of differences in the minor fragments were seen. The stability but a number of differences in the minor fragments were seen. The stability of the histone locus restriction pattern was tested by following the DNA derived from a single second chromosome of the b Adhn2 pr cn strain over a two year period. The restriction pattern of major and minor bands remained identical. Finally, histone loci distinguishable by their restriction pattern on blots were recombined with visible markers. These chromosome will be useful in tracing the fate of specific histone loci during genetic manipulations. PMID- 6290153 TI - [A preliminary study on the benzo(a)pyrene content of pastries baked with various heat sources]. PMID- 6290154 TI - [Anesthetic management of insulinoma: experiences in 76 cases]. PMID- 6290155 TI - [Study of the metabolism of cyclic AMP in human myometrium at the end of pregnancy]. PMID- 6290156 TI - Perinatal vitamin D metabolism. PMID- 6290157 TI - Lack of effect of refining on the glycemic response to cereals. PMID- 6290158 TI - Nutritional management of pregnancy complicated by diabetes: historical perspective. AB - The important role of diet in the management of pregnancies complicated by diabetes has been recognized since the nineteenth century. In this historical review we have traced the evolution of the diabetic diet from the pre-insulin era, when diabetic pregnancies were not only rare, but accompanied by high maternal mortality and fetal loss to 1981, when the nutritional management of carbohydrate intolerance during gestation is again raising provocative questions. Our recent understanding of diabetes as a heterogeneous syndrome, the 1979 dietary guidelines of the American Diabetes Association (ADA), and the 1980 revised Recommended Dietary Allowances (RDA) for pregnant and lactating women are summarized as representative of the current approach to the nutritional management of diabetic pregnancies. Many questions remain unanswered. These include the hundred-year-old debate concerning the optimal amount of carbohydrate in the diabetic diet, the possible beneficial role of large amounts of dietary fiber, and the nagging concern about total caloric intake in type I insulin dependent diabetes mellitus (IDDM) versus type II non-insulin-dependent diabetes mellitus (NIDDM) pregnant diabetic women. We suggest that nutritional counseling during gestation in the future may need to be more highly individualized as metabolic distinctions between different types of patients with carbohydrate intolerance are more clearly delineated. PMID- 6290159 TI - [Clinical and pathological study of Paget's disease of the breast--a report of 40 cases]. PMID- 6290160 TI - [Characteristics of the ultrastructural changes in the synapses of hippocampal field CA-3 in different functional states in vitro]. PMID- 6290161 TI - [Secondary integration of excised transposons]. PMID- 6290162 TI - [Inhibition of the translocation of ampicillin transposon Tn1 by the F plasmid of Escherichia coli K-12]. PMID- 6290163 TI - [Role of direct and inverted repetitions in the transformation of Bacillus subtilis by plasmid DNA]. PMID- 6290164 TI - [Lipid dependence of nuclear glucose-6-phosphatase in the tumor growth process and the change in its nature after the irradiation of tumor carriers]. PMID- 6290165 TI - [Plasmid vectors containing early promotors of phage lambda]. PMID- 6290166 TI - Oxidative metabolism of polycyclic aromatic hydrocarbons to ultimate carcinogens. PMID- 6290167 TI - [GRID syndrome]. PMID- 6290168 TI - [Prevention of central nervous system involvement with intrathecal 198Au colloid and methotrexate in non-Hodgkin lymphoma, acute non-lymphatic leukaemia and Ewing's sarcoma]. PMID- 6290169 TI - [Sodiumbicarbonate and glucose treatment for asphyxial calves]. PMID- 6290170 TI - [Aujeszky's disease of swine--demonstration of specific antibodies in organs with ELISA]. PMID- 6290171 TI - Development of beta-adrenergic receptors in normal and mutant mouse cerebellum. AB - [3H]-dihydroalprenolol was used to examine the development of beta-adrenoceptors in the cerebellum of weaver, reeler, staggerer, and jimpy neurologically mutant mice and their normal counterparts. In normal animals the greatest increase in [3H] binding occurred during the postnatal interval of 10-15 days, and maximum values were obtained at about 3 weeks. Binding was saturable with an apparent dissociation constant of 0.26 nM, and the affinity of [3]-dihydroalprenolol for its receptor did not change significantly during development. [3H] dihydroalprenolol binding was significantly reduced in weaver, reeler, and staggerer (14-22% of control values) at 20 days, but not in the jimpy mutant. These results are discussed in relation to the ontogeny of beta-adrenoreceptors with the known noradrenergic innervation of the cerebellum, and to the paucity of both granule neurons and myelination which occurs in the neurological mutants. PMID- 6290172 TI - Regional distribution of perchloric acid soluble proteins during the development of rat brain. AB - The regional contents of perchloric acid (PCA)-soluble proteins were determined during the development of rat brain. The soluble proteins in cortex, mesencephalon and medulla oblongata decreased gradually to adult age, whereas the levels in cerebellum increased significantly 12 days after birth followed by a decline to adult age. The developmental changes in the soluble proteins in retina, which showed the highest content of these proteins, were different from that of the other regions. Changes in the staining intensities of the main protein bands following SDS-gel electrophoresis were observed during the development of all regions. The patterns of the soluble proteins from retina and cerebellum were less complex than those of the other brain regions and liver and resembled that of the chromosomal high-mobility group proteins and H 1 histones. PMID- 6290173 TI - Development of the mammalian pineal gland. AB - The literature concerning the morphological, biochemical, and pharmacological development of the mammalian pineal gland is reviewed with an emphasis on the ontogeny of pineal innervation by noradrenergic neurons. Mature pineal responses to various pharmacological manipulations and the well-characterized rhythms in pineal biochemistry appear only after postnatal axogenesis of sympathetic neurons, synaptogenesis between sympathetic nerve terminals and pineal adrenergic receptors, and pineal nerve terminal synthesis, storage, and inactivation of catecholamine neurotransmitter. Since postsynaptic biochemical capacities for pineal enzymatic and endocrine function appear at a relatively early age, it is proposed that development of presynaptic inputs to the pineal is rate-limiting in the maturation of the pineal gland as a circadian neuroendocrine transducer. Because of this, the pineal gland is an excellent model for the functional development of noradrenergic inputs to selective target organs. PMID- 6290174 TI - Postnatal development of GABA binding sites and their endogenous inhibitors in rat brain. AB - Developmental changes in the affinity and density of binding sites for the inhibitory neurotransmitter, GABA, were studied in well-washed, frozen-thawed or Triton detergent extracted forebrain and cerebellar membranes from rats of 1 day, 10 days and 12 weeks postnatal age. Two populations of binding sites with similar affinities to those found in adult brain membrane preparations were found in the corresponding preparations from 1- and 10-day-old animals. While there appeared to be little developmental change in binding affinities, the density of GABA binding sites increased markedly with increasing age. At each age, freeze-thawing or Triton extraction caused successive increases in the affinity of GABA binding found in washed synaptosomal membranes. This indicates the presence of endogenous inhibitors of GABA binding in rat forebrain and cerebellum before most synapses have formed. Furthermore, there appeared to be greater endogenous inhibition of GABA binding in neonatal rat brain than in the adult, suggesting that decreases in endogenous inhibitor levels during maturation may be related to the development of synaptic activity. PMID- 6290175 TI - Ontogeny of influence of clonidine on high potassium-induced release of noradrenaline and specific [3H]clonidine binding in the rat brain cortex. AB - A high (20 mM) K+-induced, Ca2+-dependent release of L-[3H]-noradrenaline (NA) from cerebral cortex slices was detected in rats on day 18 of gestation. Depolarization-evoked release (% of total radioactivity) increased with age between day 18 of gestation and day 70 of postnatal life. Clonidine (0.1 microM) significantly (p less than 0.05) reduced a high K+-induced L-[3H]NA release on days 7 and 70 but not on day 1. Both KD and Bmax of specific [3H]clonidine binding to cerebral cortex membranes rapidly increased between days 1 and 7, followed by a gradual increase to the adult level. Presynaptic alpha 2 adrenoceptors which regulate NA release seems to become sensitive to alpha 2 agonists and to reach functional maturity between days 1 and 7 after birth in the cerebral cortex of rats. PMID- 6290176 TI - Developmental patterns of three X chromosomal enzyme activities in the brains of female and male mice. Lack of sex-dependent reaction to a fractionated prenatal x ray dose. AB - The developmental profiles of the activities of the X chromosome-linked enzymes alpha-galactosidase, hypoxanthine guanine phosphoribosyltransferase, and glucose 6-phosphate dehydrogenase were studied in the brains of mice. X chromosome inactivation, which takes place at an early stage of development, led to identical enzyme activities in females and males between the 15th day of gestation and the 64th day after birth. The enzyme activities were also studied after an X-ray dose of 3 times 1.05 Gy, delivered on gestational days 11, 12 and 13. Treatment with this dose, but not with 3 times 0.95 Gy or 1.15 Gy, respectively, was followed by the death of predominantly female offspring within 2 days after birth. The assumption that the reason for this might be a reactivation of inactive female X chromosomal genes in brain cells was tested in the present experiment. There were marked radiation-induced alterations of the activities of the above-mentioned enzymes. However, sex differences of these enzyme activities were not found after the irradiation treatment. A participation of X chromosome reactivation in the elevated mortality of female offspring is therefore unlikely. PMID- 6290177 TI - Benzodiazepine receptor development in cultures of fetal mouse cerebral cortex mimics its development in vivo. AB - Using a recently developed assay method which maintains cell integrity, we have performed a developmental study of the benzodiazepine (BDZ) receptor on cells of fetal mouse cortex in cultures which were either 'neuron-poor' or 'neuron-rich'. The developmental profile of the BDZ receptor in culture, when assayed on the intact cell, closely mimicked its development in vivo, reaching maximum values at 35-42 days after conception. Further, the time course of development paralleled that reported for GABA neurons both in vivo and in vitro. We have also shown that the relative delay in development of the clonazepam-displaceable portion of BDZ binding approximated a similar delay in the reported development of glutamic acid decarboxylase activity, both parameters presumably reflecting specific neuronal development. Thus, the ontogenesis of specific BDZ receptor binding in culture bears marked similarity to the time course of development of the GABAergic system. PMID- 6290178 TI - An in situ assay for determination of benzodiazepine binding. AB - Benzodiazepine (BDZ) receptors have been demonstrated recently in a variety of mammalian tissues. However, assay methods for such receptors have required that disrupted tissues be used. We have developed an in situ assay for this receptor utilizing intact cells cultured from the cerebral cortices of fetal mice which is more sensitive and physiologic than those used previously. Results obtained with this assay differ in the following ways from those in which disrupted tissues are used: (1) total and specific BDZ binding was as much as 10-fold higher in the in situ assays; (2) Scatchard analysis of the binding data is consistently nonlinear, revealing at least two binding sites with KD values of 5.5 and 303 nM, and (3) presumed nonneuronal receptors were found in abundance. PMID- 6290179 TI - Metabolic fate of [U-14C]-labeled glutamate in primary cultures of mouse astrocytes as a function of development. AB - The metabolic fate of [U-14C]-labeled glutamate in astrocytes grown in primary cultures for 1-3 weeks in the absence or presence of dibutyryl cyclic AMP was followed using dansylation with 3H-dansyl chloride and subsequent thin layer chromatography of the dansylated amino acids. No indication was found of classical metabolic compartmentation, i.e., the specific radioactivity of glutamine (14C/3H ratio) never exceeded that of its precursor, glutamate. In accordance with a relatively late maturation of glutamine synthetase activity the rate of formation of labeled glutamine was much faster in 3-week-old than in 1 week-old cultures. The opposite was found for aspartate formation, but under all conditions incorporation of radioactivity into aspartate was pronounced. PMID- 6290180 TI - Some haematological observations of Kenyan hepatocellular carcinoma (HCC) patients. PMID- 6290181 TI - Effects of human chorionic gonadotropin and progesterone on follicular development in the immature rat. PMID- 6290182 TI - Acute inhibition of corticosteroidogenesis by inhibitors of calmodulin action. AB - To identify the possible role of calmodulin in ACTH function, we tested the ability of chlorpromazine (CP) and other calmodulin antagonists to inhibit steroidogenesis of isolated adrenocortical cells of the rat. CP reversibly inhibited maximal ACTH-induced corticosterone (B) production. The presence of the drug did not alter the ED50 of ACTH stimulation (3.2 X 10(3) pg/ml), suggesting that it inhibited ACTH-induced steroidogenesis in a noncompetitive manner. The CP concentration required for half-maximal inhibition was 8.2 microM, a value close to the dissociation constant of the CP-calmodulin complex (5.3 microM). Concentrations greater than 40 microM resulted in complete inhibition. Similar concentrations of CP inhibited ACTH-induced cAMP accumulation in a dose-dependent manner, indicating an effect of the drug on early events in ACTH action. In addition, CP also apparently acted at a site distal to the point of cAMP formation, as shown by the finding that it inhibited cAMP-induced B production. CP inhibition of ACTH-induced B production was independent of the Ca2+ concentration, suggesting that the drug did not compete with Ca2+ directly. Concentrations of CP greater than 20 microM inhibited protein synthesis as measured by leucine incorporation into cellular proteins. Thus, although the inhibitory effect of high concentrations of CP on steroidogenesis might be explained by an effect on protein synthesis, the inhibition seen at 10 microM appeared to be independent of protein synthesis. Other antagonists of calmodulin action inhibited maximal ACTH-induced B production with the following relative potencies: trifluoperazine greater than CP greater than haloperidol greater than chlordiazepoxide. This order is similar to that reported for inhibition of calmodulin-activated phosphodiesterase and for binding to calmodulin. These findings suggest that calmodulin may modulate the effect of ACTH on steroidogenesis at multiple sites. PMID- 6290183 TI - 3,5,3'-Triiodothyronine-induced synthesis of rat liver phosphoenolpyruvate carboxykinase. AB - Recent studies have indicated that the starvation induced increase in hepatic phosphoenolpyruvate carboxykinase (PEPck; EC 4.1.1.32) is accelerated in hyperthyroid animals, whereas enzyme degradation is unaffected by the thyroid state. Therefore, the present study was undertaken to investigate the possible direct effect of T3 on the synthesis of this important gluconeogenic regulatory enzyme in vivo and in the isolated perfused liver. T3 injection in hypothyroid animals stimulated PEPck synthesis within 6-12 h. The effect, being dose dependent and significant for 0.1 microgram T3/100 g BW, could be demonstrated in animals fasted or fed a carbohydrate-rich diet. Although varying in the basal rate of synthesis, the T3-induced increase in PEPck synthesis was similar in intact, thyroidectomized, adrenalectomized, and hypophysectomized animals. No additive effect with glucocorticoids was observed, suggesting that endogenous glucocorticoids are not necessary for the hormone action. The T3-induced effect on PEPck synthesis was not mediated by alterations in the endogenous cAMP level, as was indicated (1) by the different time course of PEPck induction via (Bu)2cAMP or T3, and 2) by the finding that T3 was effective also in diabetic animals, despite maximally enhanced tissue cAMP levels. In these animals insulin antagonized the T3 action on the enzyme. T3-mediated PEPck synthesis was not prevented by propranolol. Conversely, an additive effect with isoproterenol on enzyme activity was observed. T3 (1 nM) added to the isolated liver of hypothyroid rats perfused with the synthetic fluorocarbon medium supplemented with 10% iodothyronine free serum, stimulated incorporation of labeled leucine into PEPck protein within a 6-h perfusion time. Taken together, our data demonstrate that T3 at a physiological dose stimulates hepatic PEPck synthesis. PMID- 6290184 TI - D-Ala2-Met-enkephalinamide, a potent opioid peptide, alters pituitary adrenocortical secretion in rats. AB - This study was designed to examine in rats the effects of systemic administration of a potent met-enkephalin analog, D-Ala2-met-enkephalinamide (DALA), on plasma ACTH and corticosterone secretion under basal conditions. Plasma radioimmunoassayable ACTH and corticosterone showed biphasic responses to intraarterial administration of DALA in chronically cannulated, conscious, freely moving, nonstressed animals. Both plasma ACTH and corticosterone increased at 5 10 min and decreased at 45-65 min after the administration of DALA (500 micrograms/kg) compared with either the basal concentrations or the corresponding responses in saline vehicle-injected control rats. Pretreatment with the specific opioid receptor antagonist naltrexone (2 mg/kg, intraarterially) completely prevented the DALA-induced increase in the plasma ACTH concentration and blunted the DALA-induced increase in the plasma corticosterone concentration. Naltrexone also increased plasma concentrations of ACTH and corticosterone above basal levels at a later time in DALA-treated animals. The DALA-induced increase in plasma corticosterone appears to be mediated by ACTH, mice DALA failed to increase plasma corticosterone in either hypophysectomized or dexamethasone suppressed rats. DALA decreased the adrenocortical responsiveness to ACTH in hypophysectomized, but not in untreated or dexamethasone-suppressed, rats. Naltrexone increased the adrenocortical response to ACTH in DALA-treated rats. These data suggest that endogenous opioid peptides regulate pituitary adrenocortical secretion under basal conditions by modifying both ACTH secretion and the glucocorticoid response to ACTH. PMID- 6290185 TI - Effects of preincubation on the ability of rat adipocytes to bind and respond to growth hormone. PMID- 6290186 TI - Coupling of the canine renal parathyroid hormone receptor to adenylate cyclase: modulation by guanyl nucleotides and N-ethylmaleimide. PMID- 6290187 TI - Influence of hypothyroidism on growth hormone binding by rat liver. AB - The binding of 125I-iodinated human GH ([125I]iodo-hGH) to crude, membrane-rich preparations from the livers of thyroidectomized male rats was examined to determine if alterations in GH binding might explain hypothyroid-induced growth failure. Membrane preparations from chronically hypothyroid rats bound nearly twice as much labeled hGH (12.8 +/- 2.5%; mean +/- 1 SD) as those from normal rats (6.6 +/- 1.4%) or thyroxine-treated thyroidectomized animals (7.0 +/- 1.1%). Binding by membrane preparations from hypothyroid rats treated with hGH for 2 weeks was 12.3%. By Scatchard analysis the apparent affinities of the membrane preparations for hGH were relatively constant (Ka = 2.30-2.88 X 10(9) M-1) among the experimental groups. Ovine PRL was only 1.4% as potent as hGH in displacing [125I]iodo-hGH from the liver preparations, indicating that hGH was bound primarily to somatogenic sites. Since administration of hGH did not reduce the binding of [125I]iodo-hGH, it is unlikely that the increase in hGH binding in hypothyroidism is mediated by a reduction in the ambient GH concentration. Furthermore, this increase in [125I]iodo-hGH binding indicates that an alteration in binding of GH to its receptor probably does not mediate either the low somatomedin levels or the growth failure that result from hypothyroidism. PMID- 6290188 TI - Proopiocortin-converting enzyme activity in bovine neurosecretory granules. AB - Neurosecretory granules (NSGs) from neural lobes of bovine pituitary glands were isolated in a highly purified form by metrizamide-sucrose gradient centrifugation. The purified NSGs were lysed and centrifuged, and the supernatants were further fractionated by gel filtration on Sephadex G-75. Proopiocortin-converting enzyme activity was assayed by incubation of [3H]arginine- or [3H]phenylalanine-labeled toad proopiocortin with NSG supernatant fractions. The processed products were identified by immunoprecipitation with ACTH and beta-endorphin antisera, followed by acid-urea gel electrophoresis. The optimum pH for the enzyme-mediated conversion was around pH 5.0. Conversion of toad proopiocortin by NSG converting enzyme activity was inhibited by leupeptin, antipain, p-chloromercuribenzoate, and pepstatin A, but not by diisopropyl fluorophosphate, EDTA, or N-alpha-p-tosyl-L-lysine chloromethyl ketone HCl. The results suggest that the proopiocortin-converting enzyme activity in bovine neurosecretory granules is due to an acid-thiol protease which may contain secondary hydrophobic binding sites that are involved in substrate recognition. PMID- 6290189 TI - In vitro and in vivo evidence for an indirect mechanism mediating enhanced aldosterone secretion by metoclopramide. AB - Metoclopramide (MCP), a dopaminergic antagonist, increases the levels of plasma aldosterone in man and sheep. The present studies were designed to determine how MCP exerts this effect. In in vitro studies using collagenase-dispersed rabbit adrenal zona glomerulosa cells, MCP (10(-4) M) failed to increase aldosterone biosynthesis and had no effect on the dose-related increases induced by angiotensin II (AII) or ACTH. Dopamine (10(-5) M) had no effect on the AII- or ACTH-induced aldosterone responses of these cells. Aldosterone production of cells pretreated with dopamine and stimulated by AII or ACTH was unaltered by the addition of MCP. Bolus intraarterial injections of MCP increased plasma aldosterone significantly; however, this response was completely abolished by concomitant administration of L-dopa. Chronic im administration of MCP produced significant elevations of plasma aldosterone that were associated with increases in adrenal weight and in the adrenal weight to body weight ratio. Glomerulosa cells isolated from these adrenal glands had normal basal aldosterone production and exhibited enhanced sensitivity to AII but normal responses to ACTH. These results suggest that MCP is devoid of intrinsic steroidogenic activity and that it increases aldosterone production by antagonizing a tonic inhibitory dopaminergic mechanism that leads to enhanced aldosterone production. This enhanced aldosterone production is mediated in part by increased adrenocortical sensitivity to AII. PMID- 6290190 TI - Effects of cytochalasin B on unstimulated and adrenocorticotropin-stimulated adrenocortical tumor cells in vitro. PMID- 6290191 TI - Dose-related influence of indomethacin on parathyroid hormone-stimulated adenosine 3',5'-monophosphate release from the perfused rat hindlimb. AB - We examined the effect of indomethacin (INDO) on PTH-stimulated cAMP release from the perfused rat hindlimb. Since this preparation has not been used previously to study the effects of PTH, we first determined the dose-response curve for cAMP release in response to the 1-34 fragment of synthetic bovine PTH. cAMP release peaked 3-6 min after the PTH bolus and declined gradually toward baseline, even with sustained PTH infusion. The rate of cAMP release was directly related to the PTH dose. The lowest PTH priming dose that provoked a significant increase in cAMP release was 0.6 IU. Maximal cAMP release, occurring in response to a PTH priming dose of 30 IU, was 3- to 4-fold greater than baseline. PTH caused no increase in cAMP release from or the cAMP content of isolated skeletal muscle in vitro, suggesting that cAMP released from the hindlimb in response to PTH is derived solely from bone. PTH-stimulated cAMP release was unaltered by pretreatment of the intact rat with 2 mg/kg INDO, a dose that blocks prostaglandin synthesis. In contrast, PTH-stimulated cAMP release was significantly attenuated by pretreatment with 75 mg/kg INDO. The effect was not dependent on the addition of drug to the perfusate and was not altered by thyroparathyroidectomy at the time of INDO administration. We conclude that 1) the perfused rat hindlimb can be used to examine PTH effects on bone; 2) 2 mg/kg INDO has no effect on PTH-stimulated cAMP release from the perfused rat hindlimb; and 3) INDO in high doses blunts PTH activation of adenylate cyclase. PMID- 6290192 TI - Paradoxical response to parathyroid hormone of renal handling of phosphate in hyperparathyroid rats. AB - The effect of PTH given as a bolus on renal handling of phosphate in rats was studied by a perfusion balance study method. In vitamin D-fed rats, iv PTH as a bolus (10 USP units) elicited a phosphaturic response and an increase in urinary cAMP, whether or not the rats were thyroparathyroidectomized. When given to thyroparathyroidectomized rats receiving continuous infusion of PTH (2.5 USP units/h) for 16 h, PTH led to a paradoxical decline of phosphate clearance in spite of an increase in urinary cAMP. Vitamin D-deficient intact rats also showed this paradoxical response. Vitamin D-deficient thyroparathyroidectomized rats showed an ordinary phosphaturic response. When PTH had been supplied continuously (2.5 USP units/h) to these rats, the PTH bolus again produced the paradoxical response. Phosphate supplementation did not restore the phosphaturic response. (Bu)2cAMP infusion reproduced the paradoxical effect in vitamin D-fed, thyroparathyroidectomized rats receiving continuous infusion of PTH. These data indicate that a long-lasting excess of circulating PTH alters the nature of the response of the kidney to an iv bolus of PTH, so that PTH leads to a paradoxical decline of phosphate clearance, and that the mechanism responsible for the paradox may lie in a step (or steps) subsequent to cAMP production. PMID- 6290193 TI - Effects of cyclic nucleotides on estradiol binding in human endometrium. AB - The addition of molybdate to intact or homogenized cells of the endometrial adenocarcinoma line HEC-1 during incubation with [3H]estradiol ([3H]E2) at 4 C causes substantial increases in cytoplasmic E2 binding levels. A similar effect can be observed in homogenates of normal human endometrium. These effects of molybdate appear to involve activation of E2-binding sites. Fractionation of the homogenates and recombination of different fractions revealed that activation of specific E2-binding sites by by MoO4= requires cytosolic factors as well as factors associated with the cell membrane. In homogenates of neoplastic cells (HEC-1) and normal endometrium, the addition of ATP, GTP, or cGMP was also found to increase E2 binding to levels as high as those obtained by the addition of MoO4=. In contrast, the addition of cAMP was found to lower specific E2 binding levels and to counteract the effects of MoO4=, ATP, GTP, and cGMP. Levels of intracellular cAMP and cGMP can change rapidly in cells in culture. Since cGMP causes E2 binding levels to increase while cAMP causes them to decrease, changes in the levels of these two cyclic nucleotides may explain the fluctuation in concentrations of specific estrogen binders that we have previously reported to occur in cultured endometrial cells. PMID- 6290194 TI - Extraction of parathyroid hormone and release of adenosine 3',5'-monophosphate by isolated perfused bones obtained from dogs with acute uremia. PMID- 6290195 TI - The mechanism of mineralocorticoid action of carbenoxolone. AB - The principal side effects of the drug carbenoxolone (Biogastrone; 18 beta glycyrrhetinic acid sodium hemisuccinate) are sodium retention, hypokalemic alkalosis, suppressed plasma renin, and hypertension. In previous animal studies, carbenoxolone appeared not to have intrinsic mineralocorticoid activity but, rather, to enhance aldosterone action by displacing it from nonspecific binding sites. We here report studies showing that carbenoxolone has demonstrable affinity for rat kidney mineralocorticoid receptors, intrinsic mineralocorticoid activity in the adrenalectomized rat at doses consistent with its receptor affinity, and, in addition, a powerful action of amplifying the electrolyte effects of near-maximal doses of aldosterone. PMID- 6290196 TI - Brain receptors for blood-borne calcitonin in rats: circumventricular localization and vasopressin-resistant deficiency in hereditary diabetes insipidus. AB - Specific binding sites for blood-borne calcitonin were localized by means of quantitative radioautography to the circumventricular organs of the rat brain. By this method, using normal Long-Evans rats as controls, specific binding of blood borne calcitonin in the median eminence region of the hypothalamus was reduced by one-third in homozygous Brattleboro rats, which are genetically deficient in vasopressin. Competitive binding analysis in vitro of the hypothalami from these animals confirmed the binding deficit in homozygous rats, and Scatchard analysis suggested a reduction in the number of binding sites. In homozygous rats daily vasopressin replacement therapy restored normal water balance but did not normalize the hypothalamic calcitonin binding deficit. These studies delineate for the first time specific sites within the central nervous system which could serve to mediate direct actions of blood-borne calcitonin on brain function. The deficit in the Brattleboro rat may provide a model for further investigation of the role of calcitonin within selective regions of the central nervous system. PMID- 6290197 TI - Acquisition of regulatory mechanisms for gonadotropin receptors and steroidogenesis in the maturing rat testis. PMID- 6290198 TI - Vasopressin potentiates cyclic AMP accumulation and ACTH release induced by corticotropin-releasing factor (CRF) in rat anterior pituitary cells in culture. PMID- 6290199 TI - Photoaffinity labelling of the rat liver nuclear thyroid hormone receptor with [125I]triiodothyronine. AB - [125I]Triiodothyronine (T3) was used as a photoreactive probe for the thyroid hormone nuclear receptor in photoaffinity labelling experiments. Autoradiograms of photolysis products electrophoresed on either one or two-dimensional gels showed that [125I]T3 covalently, but nonspecifically, labelled many proteins in the partially purified receptor preparations used. However, one of these proteins with an estimated molecular weight of 47,000 and an isoelectric point of approximately 6.2 +/- 0.5 pH units appears to be the thyroid hormone receptor, since, in contrast to the other proteins, its photoinduced labelling was blocked by concentrations of T3 and thyroxine (T4) similar to those that inhibit binding of [125I]T3 by the receptor in equilibrium binding assays. In addition, the isoelectric point of the photolabelled protein agrees with that determined in separate equilibrium isoelectric focusing studies. These results indicate that [125I]T3 can serve as a photoreactive probe for the thyroid hormone nuclear receptor, and they suggest that this receptor is a single polypeptide chain of molecular weight 47,000 with an isoelectric point of 6.2 +/- 0.5 pH units. PMID- 6290200 TI - A case with isolated ACTH deficiency accompanying chronic thyroiditis. AB - An unusual case of isolated ACTH deficiency with coexisting chronic thyroiditis in a 53-year-old man is reported. The patient was admitted with a 2-year history of generalized fatigue, a 13-kg weight loss, muscular weakness, and frequent hypotensive and hypoglycemic attacks. On admission serum thyroxine and triiodothyronine were significantly elevated. Basal TSH concentration was not detected and TSH showed no response to TRH, but one month after replacement therapy with hydrocortisone it was shown that serum T3, T4 and TSH response were all within normal limits. Thyroid antibodies were positive and biopsy of the thyroid gland showed chronic thyroiditis. Arginine and 1-Dopa provoked a subnormal rise in GH with a maximum of 5.6 ng/ml and 5.0, respectively. One month after treatment with hydrocortisone, GH response to 1-Dopa and arginine increased to the normal range. Prolactin response to TRH was normal and FSH response to LHRH was also normal. LH showed an exaggerated response to LHRH, although a normal response was revealed after treatment with hydrocortisone. We also presented a summary of 44 Japanese cases, 23 males (mean age; 46 yrs old) and 21 females (mean age; 48 yrs old), with isolated ACTH deficiency. PMID- 6290201 TI - Absence of response of chick embryonic limb to the growth stimulatory effect of parathyroid hormone in vitro after exposure to 6-aminonicotinamide in ovo. AB - The biochemical effect of a nicotinamide analogue, 6-aminonicotinamide (6-AN), on developing chick embryonic femur was studied. Growth of femur from 9-day-old embryos that had been exposed to 6-AN for 5 days in ovo was not stimulated by PTH in an in vitro culture system. PTH caused a much smaller increase in the cyclic AMP content in 6-AN-treated femur than in control femur. However, dibutyryl cyclic-AMP stimulated growth of 6-AN-treated femur. A defect in response of 6-AN treated femur to the growth stimulating action of PTH may explain the production of micromelia by 6-AN. PMID- 6290202 TI - Measurement of human FSH by radioreceptor assay. AB - We established a sensitive RRA system for human FSH, employing PMS-treated immature rat ovary. The dissociation constant of the binding of the receptor preparation to NIAMDD human FSH-2 was 1.15 x 10(-10) M. The standard curve was obtained with 0.2-25.6 ng/tube of NIAMDD hFSH-2. Purified hLH, hTSH, and HCG had no significant effect on the binding. When the anterior pituitary homogenates obtained from humans were assayed by this system, the intra-assay and inter-assay coefficients of variation were 11.9% and 13.4% respectively, and the assay values correlated well with those obtained by RIA. This assay is applicable for the measurement of FSH in serum, when the non-specific inhibitor effects of serum are compensated for by the addition of merthiolate and when FSH-free serum is used instead of the buffer for the standard curve. The intra-assay and inter-assay coefficients of variation were 9.31% and 19.7% respectively. The assay values correlated with those obtained by RIA under the same physiological state. The ratio of the assay values RRA/RIA, was dependent upon the physiological state, e.g. 6.29 in men, 3.84 and 4.18 in women at follicular and luteal phase respectively and 2.40 in menopausal women. During the menstrual cycle, our results showed that the value of RRA/RIA derived from serum did not change significantly. PMID- 6290203 TI - In vitro effect of LH-releasing factor on the content of cyclic cytidine 3' 5' monophosphate (c-CMP) in the rat anterior pituitary. AB - The effect of LH-RF on pituitary content of cyclic cytidine 3' 5' monophosphate (c-CMP), one of the newly detected cyclic nucleotides, was examined by the RIA procedure. In results showed that the pituitary content of c-CMP was lower than in various other organs of the rat. When incubated in vitro in the presence of 0.5 ng LH-RF/ml incubation medium, c-CMP content of the anterior pituitaries was reduced slightly, but the difference was not statistically significant when compared with the controls. No difference in c-CMP content was observed between the controls and the 5 ng LH-RF/ml group. C-CMP content in the rat anterior pituitary tissue did not change significantly during an in vitro time-course study (5, 15, and 30 min.) in the presence of 5 ng LH-RF/ml. In contrast, c-AMP content of the pituitary was significantly (P less than 0.05) elevated by the stimulation of 5 ng of LH-RF/ml at a 15 minutes of incubation. These data suggest that c-CMP content, unlike c-AMP, might not be changed significantly by hypothalamic gonadotropin releasing hormone. PMID- 6290204 TI - Effect of sodium loading and depletion on cyclic nucleotides in plasma and aorta. Interaction between prostacyclin and cyclic nucleotides. AB - The present study evaluated the effect of sodium loading and sodium depletion on cAMP and cGMP content of rat aorta. Enhanced generation of prostacyclin (PGI2) in aorta was noticed in sodium loading and sodium-depletion. As PGI2 is potent vasoactive agent, the interaction between PGI2 generation in the aorta and cyclic nucleotide accumulation in the aorta was investigated. Sodium depletion of rats induced the elevation of both cAMP and cGMP in the aorta and of cAMP in plasma. No change in cyclic nucleotides was noticed following sodium loading. These changes in cyclic nucleotides were felt to reflect the fluctuation of vasoactive agents under various sodium metabolism conditions. These observations may indicate that PGI2 is not a major factor in the regulation of cyclic nucleotide metabolism under sodium loading and sodium depletion. The increased accumulation of cAMP and cGMP in rat aorta in sodium depletion may be due to the cumulative effects of PGI2, catecholamine and probably angiotension II which are induced by sodium depletion. PMID- 6290205 TI - Augmentation of ACTH-induced steroidogenesis in isolated rat adrenal cells by rat tissue extracts and by hormone preparations. PMID- 6290206 TI - Stimulatory effect of pertussis toxin on tissue cyclic AMP levels in canine thyroid slices. AB - The in vitro effects of the protein purified from the culture medium of Bordetella pertussis (pertussis toxin) on cyclic AMP levels in canine thyroid slices were studied. The toxin caused a three-fold increase in tissue cyclic AMP levels after 18 h or longer incubation. Furthermore, when the slices were incubated with the toxin for 6 h and then incubated without the toxin for further 12 h, similar effects were observed. These results indicate that pertussis toxin elevates tissue cyclic AMP levels after a time lag in a manner different from other thyroid stimulators. PMID- 6290207 TI - Effects of diethyldithiocarbamate (DDC), fusaric acid, catecholaminergic and serotonergic receptor blockers on the ascending pathway from the ventrolateral part of the medulla oblongata. AB - The effect of a potent inhibitor of dopamine-beta-hydroxylase, diethyldithiocarbamate (DDC) or fusaric acid, on the antidromically evoked potentials by electrical stimulation of the preoptic suprachiasmatic area (POSC) was examined. Thirty seven neurons in the ventrolateral part of medulla oblongata (VLMO) were successfully tested after DDC injection. Nineteen of them decreased in amplitude to approximately 50% of control levels at 120 min after DDC injection, 20% at 180 min. These 19 neurons were characterized by having a notch in the rising phase and taking a longer time to complete the initial deflection. After fusaric acid injection, ten neurons in the VLMO were successfully tested and four of them decreased in amplitude similar to DDC injection. Effects of receptor blockers of noradrenaline, dopamine and serotonin on the single unit responses of the POSC neurons to the VLMO stimulation in proestrous rats were examined in order to know which of noradrenaline, dopamine or serotonin are involved in the neural transmission from the VLMO to the POSC. After the injection of phenoxybenzamine, an alpha-adrenergic receptor blocker, the rates of facilitatory and inhibitory responses in the POSC neurons to the VLMO stimulation were significantly decreased. Percentages of facilitatory and inhibitory responses were not changed significantly after injection of pimozide, a dopaminergic receptor blocker, or methysergide, a serotonergic receptor blocker. These findings suggest that noradrenaline might be involved in the transmission of the VLMO stimulation effect on the POSC neurons and that VLMO noradrenaline containing neurons send their axons directly to the POSC. PMID- 6290208 TI - The effect of high level CaCi2 injection on parathyroid hormone dependent cAMP in stone formation patients. AB - Patients with a history of idiopathic calcium oxalate stones but without current stone formation do not react to stimulation of parathyroid extract as expected after high dose calcium pretreatment. With Ca2+ pretreatment, these patients show higher serum and also renally generated levels of cAMP after rapid injection of extreme parathyroid extract-concentrations. Healthy controls show a modest increase of serum levels cAMP after similar stimulation with parathyroid extracts. In contrast to patients, calcium pretreatment of controls significantly decreases the renal cAMP response to parathyroid extract stimulation. Without Ca2+ pretreatment, both patients and controls show a strong increase in urinary and also in renally generated cAMP levels following parathyroid extract stimulation. PMID- 6290209 TI - Rotational motion of cytochrome c derivatives bound to membranes measured by fluorescence and phosphorescence anisotropy. AB - Molecular motion of metal-free and metal-substituted cytochrome c derivatives was examined using the anisotropy of emissions from the singlet and the triplet states. The anisotropy of fluorescence provides a means to study the motion of cytochrome c in the nanosecond time scale, since the fluorescence lifetime of metal-free cytochrome c is around 10 ns. We find that the anisotropy of fluorescence of metal-free cytochrome c when bound to mitochondria does not decay, but when bound to phospholipids has a small component which decays independently of the rotation of the whole molecule. The use of phosphorescence extends the time scale for study into the millisecond regime, since the lifetime of the excited triplet state of zinc cytochrome c, as measured by triplet-triplet absorption and phosphorescence emission is approximately equal to 9 ms for free zinc cytochrome c and 7 ms for mitochondrial membrane-bound zinc cytochrome c at room temperature. The decay of anisotropy of phosphorescence emission of mitochondrial membrane-bound zinc cytochrome c is clearly biphasic; the fast component corresponds to a rotational relaxation time of 300 mus and the slow component with relaxation time of approximately equal to 6 ms. The slow component appears to be due to the rotation of the entire mitochondrion, whereas the fast component was interpreted to be due to the rotation of cytochrome c in a cone about a single axis perpendicular to the plane of the membrane surface. PMID- 6290210 TI - A cytochrome b/c1 complex with ubiquinol--cytochrome c2 oxidoreductase activity from Rhodopseudomonas sphaeroides GA. AB - A cytochrome b/c1 complex which catalyses the reduction of cytochrome c by ubiquinol has been isolated from Rhodopseudomonas sphaeroides GA. It contains two hemes b and substoichiometric amounts of ubiquinone-10 and of the Rieske Fe-S center per cytochrome c1, and is essentially free of reaction center and bacteriochlorophyll. The complex consists of three major polypeptides with apparent molecular masses of 40, 34 and 25 kDa. The 34-kDa polypeptide carries heme. Cytochrome c1 has a midpoint potential of 285 mV. For cytochrome b two midpoint potentials, at 50 and -60 mV, at pH 7.4, can be derived if one assumes two components of equal amount. Ubiquinol--cytochrome c oxidoreductase activity is specific for ubiquinol and bacterial cytochromes c, and is inhibited by antimycin A and 5-n-undecyl-6-hydroxy-4,7-dioxobenzothiazole. The complex shows oxidant-induced reduction of cytochrome b. PMID- 6290211 TI - A restriction map of Xenopus laevis mitochondrial DNA. AB - The mitochondrial DNA from Xenopus laevis is a 17.4 x 10(3)-base-pair circular DNA molecule. The mapping of this DNA, using 19 different restriction endonucleases is reported here. The sites are as follows: 1 for BamHI, PstI, SacI, SalI, BalI; 2 for BglII, SacII, EcoRI, ClaI, 3 for XhoI, 4 for AvaI, XbaI, PvuII, 5 for HindIII, 6 for HhaI, BclI, HpaI, 10 for AvaII and 11 for HincII. The same sites (except for one of the two ClaI sites) are observed in the molecule cloned in pBR322 DNA. The fragments corresponding to 62 cleavage sites have all been ordered and precisely located. They provide suitable conditions for further investigations connected with the study of replication and nucleotide sequence determination of this molecule. PMID- 6290212 TI - ATP synthetase (F1F0) of Escherichia coli K-12. High-yield preparation of functional F0 by hydrophobic affinity chromatography. AB - 1. The purified ATP synthetase complex (F1F0) from Escherichia coli was adsorbed to immobilized poly-(L-lysine)-deoxycholic acid. About 0.7 mg F1F0 were bound per ml of settled gel. The hydrophilic F1 part was dissociated from the complex by treatment with 7 M urea. F0 was eluted in high yield either with deoxycholate (6 mM) or taurodeoxycholate (10 mM). About 14% of the total protein bound to the column was eluted as F0, which corresponds to 64% of the total F0 in the F1F0 complex. 2. The purified F0 preparation obtained was composed of three different kinds of subunits with apparent molecular weights of 24000 (a), 19000 (b) and 8300 (c), respectively as determined by sodium dodecyl sulfate gel electrophoresis. 3. After incorporation into liposomes and the generation of a potassium diffusion potential by valinomycin, the F0 preparation mediated H+ translocation. This H+ uptake is inhibited by either dicyclohexylcarbodiimide or purified F1 ATPase. 4. Incubation of F0-containing liposomes with F1 led to the reconstitution of an ATP-driven quenching of acridine-dye fluorescence. The quenching was abolished by uncoupler and prevented by dicyclohexylcarbodiimide. PMID- 6290213 TI - Biosynthesis of mitochondrial porin and insertion into the outer mitochondrial membrane of Neurospora crassa. AB - Mitochondrial porin, the major protein of the outer mitochondrial membrane is synthesized by free cytoplasmic polysomes. The apparent molecular weight of the porin synthesized in homologous or heterologous cell-free systems is the same as that of the mature porin. Transfer in vitro of mitochondrial porin from the cytosolic fraction into the outer membrane of mitochondria could be demonstrated. Before membrane insertion, mitochondrial porin is highly sensitive to added proteinase; afterwards it is strongly protected. Binding of the precursor form to mitochondria occurs at 4 degrees C and appears to precede insertion into the membrane. Unlike transfer of many precursor proteins into or across the inner mitochondrial membrane, assembly of the porin is not dependent on an electrical potential across the inner membrane. PMID- 6290214 TI - Proton-magnetic-resonance studies on the coat protein of alfalfa mosaic virus. PMID- 6290215 TI - Source of rapidly labeled ATP tightly to non-catalytic sites on the chloroplast ATP synthetase. AB - Bound [32P]ATP is found on deenergized, washed chloroplast thylakoids which were illuminated in the presence of ADP and [32P]Pi. Tight binding of [32P]ATP occurred both during and after energization. Different classes of bound [32P]ATP were distinguished on the basis of their rates of formation, susceptibility to hexokinase and displacement by unlabeled ATP. 1. The rates of formation and discharge of the rapidly labeled tightly bound ATP class were much lower than that of ATP formation. The level of this bound ATP saturates at lower concentrations of substrates than does the rate of phosphorylation. Unlabeled ATP, present in the reaction medium, displaces the rapidly labeled tightly bound ATP without affecting the rate of phosphorylation. 2. We therefore conclude that the rapidly labeled bound ATP class does not fulfill the requirements expected for a catalytic intermediate and that the nucleotide tight binding site(s) on the ATP synthetase differ from the catalytic site(s) for ATP formation. 3. Since the rapidly labeled tightly bound [32P]ATP is not abolished by high concentrations of hexokinase, but is nevertheless displaced by exogenous ATP, we propose that tight binding of ATP to non-catalytic sites occurs via a free species of newly synthesized ATP which diffuses slowly to the medium from a space accessible to ATP but not to hexokinase. PMID- 6290216 TI - Ferredoxin from Methanosarcina barkeri: evidence for the presence of a three-iron center. AB - Methanosarcina barkeri ferredoxin was purified and characterized by electron paramagnetic resonance (EPR) and Mossbauer spectroscopy. The purification procedure included chromatographic steps on DEAE-cellulose and gel filtration. The isolated protein is unstable under aerobic conditions. The ferredoxin exhibits charge transfer bands at 283 nm and 405 nm with an absorption ratio A405/A283 = 0.73. Its molecular weight has been estimated to be 20000-22000 by gel filtration chromatography. The native ferredoxin exhibits an intense EPR signal at g = 2.02 and only a very weak g = 1.94 signal develops upon reduction with dithionite. The Mossbauer spectra of the reduced protein are characteristic of a [3Fe-3S] center. The combined EPR and Mossbauer studies show that M. barkeri ferredoxin contains only [3Fe-3S] clusters, similar to Azotobacter vinelandii Fd[Emptage, M.H., Kent, T.A., Huynh, B.H., Rawlings, J., Orme-Johnson, W.H. & Munck, M. (1980) J. Biol. Chem. 255, 1793-1796], Desulfovibrio gigas FdII [Huynh, B.H., Moura, J.J.G., Moura, I., Kent, T.A., LeGall, J., Xavier, A.V. & Munck, E. (1980) J. Biol. Chem. 255, 3242-3244] and mitochondrial beef heart aconitase [Kent, T.A., Dreyer, J.-L., Kennedy, M.C., Huynh, B.H., Emptage, M.H., Beinert, H. & Munck, E. (1982) Proc. Natl Acad. Sci. USA, 79, 1096-1100]. PMID- 6290217 TI - Complete primary structure of protein phosphatase inhibitor-1 from rabbit skeletal muscle. AB - The complete primary structure of protein phosphatase inhibitor-1 has been determined. The protein consists of a single polypeptide chain of 165 residues, molecular weight 18640. The threonine residue that must be phosphorylated for activation is at position 35 and the active cyanogen bromide peptide, CB-1, comprises residues 2-66. The N-terminal methionine is acetylated and 40% of the inhibitor-1 molecules lack the C-terminal dipeptide Ala-Val. Serine-67 is substantially phosphorylated in vivo, but this phosphoserine residue does not appear to influence the activity of inhibitor-1. PMID- 6290219 TI - Phosphoribosylpyrophosphate synthetase of Escherichia coli, Identification of a mutant enzyme. AB - From an Escherichia coli purine auxotroph a mutant defective in phosphoribosylpyrophosphate (PRib-PP) synthetase has been isolated and partially characterized. In contrast to the parental strain, the mutant was able to grow on nucleosides as purine source, whereas growth on purine bases was reduced. Kinetic analysis of the mutant PRib-PP synthetase revealed an apparent Km for ATP and ribose 5-phosphate of 1.0 mM and 240 muM respectively, compared to 60 muM and 45 muM respectively for the wild-type enzyme. ADP, which inhibits the wild-type enzyme at a concentration of 0.5 mM ribose 5-phosphate, stimulated the mutant enzyme. The activity of PRib-PP synthetase in crude extract was higher in the mutant than in the parent. When starved for purines an accumulation of PRib-PP was observed in the parent strain, while the pool decreased in the mutant. During pyrimidine starvation derepression of PRib-PP synthetase activity was observed in both strains, although to a lesser extent in the mutant. Our data suggest that the mutant harbors a mutation in the structural gene for PRib-PP synthetase. The mutation responsible for the altered PRib-PP synthetase was located in the purB hemA region at 26 min on the recalibrated linkage map. PMID- 6290218 TI - Temperature dependence of ADP/ATP translocation in mitochondria. AB - The temperature dependence of the adenine nucleotide exchange in mitochondria has been determined by employing a rapid mixing, quenching and sampling apparatus and the inhibitor quench-back exchange method. Thus the exchange is resolved down to 0.1 s. Rates are evaluated from accumulating the time-dependent progress at about 10 points. The exchange rate in liver mitochondria was determined from -10 degrees C to + 10 degrees C in the presence of 20% glycol, from 0 degrees C to 25 degrees C, and from 20 degrees C to 40 degrees C under partial inhibition by carboxyatractylate. The total range between -10 degrees C to + 40 degrees C has only one temperature break at 13 degrees C. From the Arrhenius plot between -10 degrees C to + 13 degrees C, EA approximately equal to 140 kJ and above 13 degrees C, EA approximately equal to 56 kJ is evaluated, corresponding to a Q10 of 8 and 2 respectively. In beef heart mitochondria the exchange rate was measured between 0 degrees C and 20 degrees C, and between 15 degrees C and 30 degrees C under partial inhibition with carboxyatractylate. There is a temperature break around 14 degrees C with EA approximately equal to 143 kJ between 0 degrees C and 14 degrees C and EA approximately equal to 60 kJ from 15 degrees C to 30 degrees C. The extrapolated translocation rates at 37 degrees C are 500 and 1800 mumol min-1 (g protein)-1 for rat liver and for beef heart mitochondria respectively. The temperature break is suggested to reflect a conformation change since there is no reversed break at low temperature, the temperature break changes in sonic particles and no lipid phase transition at 14 degrees C in mitochondria has been reported. PMID- 6290220 TI - Isolation of zymogen granules from rat pancreas and characterization of their membrane proteins. AB - A zymogen granule fraction has been isolated from rat pancreas, and its purity has been assessed by biochemical and morphological criteria. Specific activities of two marker enzymes, amylase and chymotrypsin, are increased by 4.6 and 5.4 fold, respectively, as compared to the homogenate. The purified fraction is devoid of detectable RNA, DNA and 5'-nucleotidase, glucose-6-phosphatase, and cytochrome c oxidase activities. Electron micrographs confirm the absence of mitochondria, lysosomes, and rough endoplasmic reticulum fragments. Zymogen granule membranes were isolated from this fraction on a sucrose gradient following lysis in alkaline buffer. Secretory contaminants were efficiently removed from the membranes as indicated by experiments in which labeled secretory proteins were added during the isolation procedure and secondly by measuring residual levels of amylase and chymotrypsin. Three enzyme activities were found in the membranes: thiamine pyrophosphatase, ATP-diphosphohydrolase, and low levels of acid phosphatase. Membrane proteins were solubilized by urea-Triton X 100 and separated in double-dimension (isoelectric focusing and sodium dodecyl sulfate-polyacrylamide gel electrophoresis). Isoelectric point and molecular weight of each protein band were determined. PMID- 6290221 TI - Lysolecithin as a modifier of induced pinocytosis in Amoeba proteus. AB - Lysolecithin was found to modify cation-induced pinocytosis in Amoeba proteus. It is shown here that lysolecithin (LPC) in the concentration range of 10(-15) to 10(-10) g/ml has the same effect on Na+ -induced pinocytosis as cAMP and a pinocytosis regulating factor (PRF) which is secreted by the amoeba. Thus, LPC activated Na+-induced pinocytosis in starved amoebae and decreased the sensitivity to the inducer in normal cells. Pinocytosis depressed by treatment with EGTA or dibucaine became normal upon addition of LPC to the inducer. These effects were also obtained with lysolecithin isolated from the amoeba. It is suggested that PRF and amoeba LPC may be closely related and that phospholipase activity of the amoeba may regulate its capacity for pinocytosis. PMID- 6290222 TI - Reduction of benzo(a)pyrene induced chromosomal aberrations by DL-alpha tocopherol. AB - The antioxidant, DL-alpha-tocopherol (vitamin E), has been demonstrated to significantly reduce the percentage of benzo(a)pyrene (BP) induced chromosomal aberrations in vitro. Chinese hamster lung (Don) and Chinese hamster ovary (CHO) cells were treated with either 1 microgram/ml or 5 micrograms/ml BP for 4 to 28 h; some cultures were treated with S9 mix activated BP. Additional cultures of Don and CHO were treated simultaneously with 100 micrograms/ml of DL-alpha tocopherol and BP. In CHO cells 1 microgram/ml non-activated BP significantly increased the chromosomal aberration percentage above the control level. Aberrations observed included breaks, gaps, fusions, rings, dicentrics, and polyploids. Chinese hamster Don cells treated with 1 microgram/ml or 5 micrograms/ml S9 mix activated BP contained significant increases in aberration percentages above the control levels. When Don cells were treated simultaneously with activated BP and DL-alpha-tocopherol for 4 h, there was a slight decrease in the total aberration frequency to less than that of cells treated with activated BP only; however, when Don cells were treated with BP and DL-alpha-tocopherol for 28 h, there was a significant reduction in the aberration percentage below that of BP-treated cells alone. Similar results have been obtained with CHO cells treated with nonactivated BP and DL-alpha-tocopherol. The results reported here provide further evidence that antioxidants may prevent the potential mutagenic and carcinogenic effects of certain polycyclic compounds. PMID- 6290223 TI - The treatment of 48 patients affected by ovarian carcinoma at stages III and IV F.I.G.O. AB - The Authors analyzed the treatment of 28 patients in stage III (FIGO) and 20 patients in stage IV affected by epithelial ovarian cancer. In these cases, surgery was based on the criterion of cytoreduction and, where possible, maximum radicality. Within a month from surgery, all patients underwent a program of combined chemotherapy (VCR + 5 FU + CTX) envisaging successive cycles, every four weeks. Minimal side effects were generally observed. 13/28 (46.4%) responses were recorded in stage III and 4/14 (28.6%) in stage IV. The highest percentage of responses, 80%, was obtained in patients with a residual tumor less than 2 cm, all belonging to stage III. Stage III and stage IV patients with a residual tumor greater than 2 cm responded in 45% and 33% of the cases respectively. Patients who underwent explorative laparotomy showed negligible responses. There was a statistically significant difference survival between stage III and stage IV patients (p less than 0.001), among patients with a similar residual tumor greater than 2 cm, but in different stages (p less than 0.05), and among stage III patients who responded or not to chemotherapy (p less than 0.001). PMID- 6290224 TI - Radionuclide ejection fraction: comparison of response to treadmill and bicycle exercise. AB - Most exercise radionuclide studies utilise the supine bicycle ergometer during imaging, although exercise on a treadmill results in greater cardiovascular stress. Twenty-three patients were studied to compare the radionuclide left ventricular ejection fraction (LVEF) estimated immediately following treadmill exercise with that obtained at peak supine bicycle exercise in patients with coronary artery disease (CAD) and patients with normal coronary arteries. In 14 patients with CAD the rest LVEF was 47 +/- 7% (mean +/- SD) by first pass and 49 +/- 10% by equilibrium blood pool techniques. Immediately following maximum treadmill exercise, 13 of the 14 patients with coronary artery disease showed either no change or a decrease in LVEF, the LVEF was 49 +/- 7% (P = NS) and 47 +/ 8% (P = NS) by first pass and equilibrium determinations respectively. In comparison the LVEF during peak supine bicycle exercise decreased significantly to 42 +/- 12% (P less than 0.01). In nine patients with angiographically normal coronary arteries the rest LVEF was 51 +/- 4% by first pass and 54 +/- 6% by equilibrium, increasing to 67 +/- 7% (P less than 0.01) and 64 +/- 7% (P less than 0.01) respectively following treadmill exercise. During peak bicycle exercise LVEF increased in the normal patients to 61 +/- 7% (P less than 0.05). These data suggest that quantitative radionuclide angiography following either maximum treadmill exercise or during peak bicycle exercise can discriminate between patients with coronary artery disease and normal subjects, although the magnitude of left ventricular functional changes are greater during bicycle stress. PMID- 6290225 TI - 67Ga scanning for assessment of disease activity and therapy decisions in pulmonary sarcoidosis in comparison to chest radiography, serum ACE and blood T lymphocytes. AB - In 60 patients with histologically proven sarcoidosis, 67Ga scanning was evaluated in terms of sensitivity and specificity for assessing disease activity and compared with chest radiography, serum ACE and blood T-lymphocytes. While 67Ga scans had the highest sensitivity (94%), the specificity was only 68%. The sensitivity of chest radiography was 80%, of serum ACE and blood T-lymphocytes 77% and 48%, respectively. While in patients with radiographical type I, 67Ga scanning, chest radiography and serum ACE had a sensitivity of 92%-100%, in patients with radiographical type II-III, only 67Ga scans had a sensitivity exceeding 90%. A 67Ga score correlated significantly with serum ACE levels (r = 0.59, P less than 0.001). After effective steroid treatment, 67Ga uptake and serum ACE activities decreased markedly. While in 25% of cases, chest radiography failed to provide reliable information, 67Ga scanning and serum ACE activities always proved useful in evaluating the course of the disease and the patient's response to steroid therapy. A negative 67Ga scan together with normal serum ACE levels seem to have a high predictive value for excluding active sarcoidosis. PMID- 6290226 TI - Scintigraphy of thoracic aortic and innominate arterial aneurysms: the value of the 99mTc-pertechnetate first pass study with subsequent in vivo labeling of red blood cell pool. PMID- 6290227 TI - The adhesiveness of normal and SV40-transformed BALB/c 3T3 cells: effects of culture density and shear rate. AB - The adhesiveness of BALB/c 3T3 cells and their SV40 virally transformed counterparts as assessed by aggregation kinetics was found to vary as a function of cell culture density and aggregation conditions. In culture, SV3T3 cells were found to have a faster growth rate and a smaller cell volume than 3T3 cells. Although both cell-types displayed increasing adhesiveness with increasing culture density, the adhesiveness of SV3T3 cells was consistently lower than that of 3T3 cells of comparable culture density when the cells were aggregated under shear rate conditions less than or equal to 45/sec. When the shear rate was increased from 90 to 450/sec, however, the aggregation profile inverted, with the 3T3 cells becoming less adherent than the SV3T3 cells. The ability of the transformed SV3T3 to remain adherent under conditions of relatively high shear may facilitate extravasation during the process of tumour spread. PMID- 6290228 TI - Effect of ceruletide on rest pain in patients with arterial insufficiency of the lower extremity. AB - The effect of ceruletide (CRL), a synthetic decapeptide analogue of cholecystokinin, on rest pain and arterial blood flow was evaluated in 8 patients with advanced, occlusive atherosclerosis of the lower extremities. CRL 1, 2, or 4 ng kg-1 or placebo were infused intravenously in random order, and in a double blind fashion. Pain relief, assessed by a scoring system, was significantly better (p less than 0.01) following the 2 and 4 ng kg-1 doses of CRL (2.71 and 2.66, respectively) than following placebo (0.75). Arterial blood flow was not affected by either CRL in any dose or by placebo. Pretreatment with naloxone, a pure opioid antagonist, abolished the analgesic effect of CRL. Following the 2 ng dose of CRL, beta-endorphin levels were significantly elevated from a basal value of 125 +/- 15 pg/ml to 191 +/- 35 pg/ml 5 h after CRL administration (p less than 0.05). Circulating levels of ACTH, prolactin and GH were not affected by CRL. It is concluded that CRL was effective in relieving ischaemic rest pain, and that the mechanism was related to the release of endogenous opioids. PMID- 6290229 TI - Elevated liver enzymes in serum during suloctidil treatment. PMID- 6290230 TI - Up-regulation of rat cortical adenosine receptors following chronic administration of theophylline. PMID- 6290231 TI - Inhibition of postsynaptic dopamine-sensitive adenylate cyclase in carp retina by the putative dopamine autoreceptor agonist, 3-PPP. PMID- 6290232 TI - High-affinity binding sites for gastrin on isolated rabbit gastric mucosal cells. AB - A biologically active gastrin analogue, [125I](Nle11)-HG-13, appears to bind specifically to saturable binding sites on isolated rabbit gastric mucosal cells: Kd = 70 pM at pH 7.4 and at 37 degrees C. Increasing incubation temperature from +4 degrees C to +37 degrees C increased specific binding. Gastrin binding was shown to be reversible and the dissociation rate was enhanced with cold gastrin. The binding sites were saturated with 0.2 fmol of labelled gastrin per 10(6) mucosal cells. Gastrin binding was not inhibited by secretin, glucagon, Met enkephalin, physalaemin, eledoisin, BPP, VIP, carbachol, histamine, atropine or cimetidine. Gastrin analogues (HG-4, HG-8, (Leu15)-HG-17), CCK-7 and gastrin antagonists (proglumide or benzotript) inhibited [125I](Nle11)-HG-13 specific binding. We concluded that isolated cells from rabbit gastric fundic mucosa contain high-affinity binding sites for a gastrin analogue (Nle11)-HG-13. PMID- 6290233 TI - Stimulation of ureogenesis through alpha 1- and beta-adrenoceptors in juvenile rat hepatocytes. PMID- 6290234 TI - Autoreceptors modify the evoked release of [3H]GABA from cerebellar neurons in dissociated cell culture. AB - We investigated the effect of GABA, muscimol and THIP on the K+ -stimulated and spontaneous release of [3H]GABA from neuron-enriched cell cultures of the rat cerebellum. Each agonist produced significant reductions in evoked [3H]GABA without causing marked changes in spontaneous release. The agonist-induced inhibition of K+ -stimulated [3H]GABA release was reversed by the GABA antagonists bicuculline and picrotoxin. It is suggested that GABAergic neurons in cerebellar cell cultures possess GABA receptors which are involved in the regulation of evoked transmitter release. PMID- 6290235 TI - Non-opioid psychotomimetic-like discriminative stimulus properties of N allylnormetazocine (SKF 10,047) in the rat. AB - The purpose of this study was to characterize the discriminative stimulus property of dl-SKF 10,047 in the rat and to investigate if this property is the result of an interaction with opiate receptors or dopaminergic mechanisms. Male Sprague Dawley rats were trained in a two-lever food-reinforced procedure to discriminate between the effect of saline and of dl-SKF 10,047 (5 mg/kg). The trained rats dose dependently generalized the effect of dl-, d- and l-SKF 10,047, as well as cyclazocine, phencyclidine and ketamine but did not generalize the effect of either fentanyl, morphine, bremazocine, ethylketocyclazocine, pentazocine, nalorphine, naloxone, MR 2266, d-lysergic acid diethylamide, apomorphine or d-amphetamine. Neither naloxone, MR 2266, fentanyl, ethylketocyclazocine nor haloperidol antagonized the discriminative stimulus produced by dl-SKF 10,047. These data suggest that the discriminative stimulus property of dl-SKF 10,047 in the rat is not due to its interaction with either opiate receptors or dopaminergic mechanisms but may be related to its psychotomimetic effect in man. PMID- 6290236 TI - Norepinephrine constricts the canine coronary bed via postsynaptic alpha 2 adrenoceptors. AB - The effect of alpha 2-blockade (0.3 mg/kg i.v. rauwolscine) and alpha 1-blockade (1.2 mg/kg i.v. prazosin) on coronary constrictions induced by intracoronary injections of azepexole (B-HT 933, alpha 2-agonist, 0.1-10 microgram/kg), phenylephrine (0.3-3 microgram/kg) and norepinephrine (0.001-0.1 microgram/kg) were studied in dog hearts perfused in situ under beta-blockade. Constrictions by azepexole (antagonized by rauwolscine, yet resistant to prazosin and methysergide) demonstrated coronary alpha 2-adrenoceptors. Norepinephrine-induced constrictions were more attenuated (22-fold) by alpha 2-blockade than by alpha 1 blockade (2.6-fold) and thus were mediated mainly by activation of postsynaptic alpha 2-receptors. PMID- 6290237 TI - Opiate receptor binding of nicomorphine and its hydrolysis products in rat brain. PMID- 6290238 TI - Does the reversal of the anticonflict effect of phenobarbital by beta-CCE and FG 7142 indicate benzodiazepine receptor-mediated anxiogenic properties? AB - In mice and rats, the high affinity ligand for brain benzodiazepine (BZ) receptors beta-CCE, and the more stable congener FG 7142, failed to exert anticonflict activity in conflict situations but instead reversed the anticonflict effect of lorazepam. In contrast to Ro 15-1788, beta-CCE and FG 7142 also antagonized the anticonflict effect of phenobarbital in rats. This effect suggests that beta-CCE and FG 7142 may produce anxiety by either inducing a conformational change in the BZ receptors which is directly opposite to that induced by the benzodiazepines, or binding to a particular subclass of BZ receptors. PMID- 6290239 TI - Differentiation of benzodiazepine receptor agonist and antagonist: sparing of [3H]benzodiazepine antagonist binding following the photolabelling of benzodiazepine receptors. PMID- 6290240 TI - VIP stimulates ACTH release and adenylate cyclase in human ACTH-secreting pituitary adenomas. AB - The effect of vasoactive intestinal polypeptide (VIP) on in vitro ACTH release and adenylate cyclase activity was investigated in human ACTH-secreting pituitary adenomas from 4 patients with Cushing's disease and 2 patients with Nelson's syndrome. In all the tumors tested, VIP elicited a dose-dependent stimulation of hormone release from adenoma fragments (90-247% at 10-7 M VIP) and of cAMP formation in membrane preparations (75-140% at 3 X 10-6 M VIP). Therefore a role of VIP in the control of ACTH secretion in human ACTH-secreting adenomas is suggested; a cAMP-dependent mechanism of action can also be hypothesized. PMID- 6290242 TI - Radioenzymatic assay measurement of yohimbine's influence on adrenergic neurotransmission of rat vas deferens: unique consequence of using normetanephrine as an uptake2-blocking agent. AB - The effect of the alpha 2-receptor antagonist, yohimbine on norepinephrine overflow was studied in the transmural-stimulated isolated rat vas deferens. A radioenzymatic assay was used to measure the endogenous norepinephrine overflow. In initial studies conducted in the presence of uptake1 blocker desipramine (10( 6) M) and uptake 2 blocker, normetanephrine (10(-5) M) there was an apparent uncoupling of the influence of yohimbine on nerve-stimulated contractile response from norepinephrine overflow. These results were found to be due to the electrolytic O-demethylation of normetanephrine with the resultant generation of large quantities of norepinephrine obscuring the influence of yohimbine on nerve stimulated norepinephrine overflow from the vas deferens. These findings serve as a warming to the use of normetanephrine as an uptake1 blocker when radioenzymatic assay is used to measure norepinephrine overflow from transmural-stimulated isolated tissue preparations. Yohimbine (10(-6) M), in the absence of uptake blockade, causes a 3-fold enhancement of nerve-stimulated norepinephrine overflow at 1 Hz and a 2-fold enhancement at 10 Hz. This report demonstrates utilization of a radioenzymatic assay to study endogenous norepinephrine overflow from rat vas deferens. Results for yohimbine are complementary to others using measurement of 3H-label overflow from [3H]norepinephrine prelabeled isolated tissue. PMID- 6290241 TI - The cyclic AMP system in sensitized and desensitized guinea-pig tracheal smooth muscle. AB - Egg albumin decreased the cyclic AMP content and the protein kinase ratio of sensitized guinea-pig tracheal smooth muscle. The effect preceded the contraction of the tracheal preparation. Antigen challenge increased cyclic AMP phosphodiesterase activity without changing the Km value. In tracheal preparations of desensitized guinea-pigs, the mechanical and metabolic effects of egg albumin were markedly reduced. These results suggest that the effects on the cyclic AMP system in response to immunological challenge might represent an important mechanism modulating the contractile response of guinea-pig tracheal smooth muscle. PMID- 6290243 TI - Effect of pertussis vaccine on alpha-adrenoceptors of the circulatory system of the rat. AB - Administration of pertussis vaccine to rats markedly diminished the sensitivity of their vascular alpha 2-adrenoceptors as evidenced by shifts to the right and decreased maximal pressor responses to clonidine and azepexole in pithed rats. No effect was observed on the sensitivity of alpha 1-adrenoceptors as evidenced by the dose-response curve to methoxamine. A slight persistent tachycardia was also produced by the vaccine. A model to explain the effects of the vaccine on the vascular system of the rat is proposed. PMID- 6290244 TI - Increased renal beta-adrenoceptors in stroke-prone spontaneously hypertensive rats. PMID- 6290245 TI - Effects of agents which enhance GABA-mediated neurotransmission on licking conflict in rats and exploration in mice. AB - On the holeboard, exploration (dipping) and locomotion of mice were enhanced by non-sedative doses of anxiolytics; clobazam, diazepam, nitrazepam and flunitrazepam. After chronic dosing the sedative effects of flunitrazepam showed tolerance and the increases in exploration remained while locomotion was less increased. Sodium valproate and to a lesser extent GAG also increased both exploration and locomotion at non-sedative doses. Muscimol, AOAA and urethane increased locomotion at threshold sedative doses with little or no increase in exploration. Baclofen only decreased behaviour. In a drinking test benzodiazepines evoked large increases in punished drinking. After chronic dosing of flunitrazepam there was a greater increase. Sodium valproate evoked a response of similar magnitude to the benzodiazepines whilst GAG evoked a smaller increase. Muscimol did not increase punished drinking. It is suggested that anxiolytic drug actions result from a more selective enhancement of brain GABA transmission. PMID- 6290246 TI - Demonstration of a slowly dissociating form of bovine hippocampal synaptic membrane opiate receptors. AB - We studied the binding characteristics of opiate receptors in synaptic plasma membranes isolated from bovine hippocampus. On the basis of kinetic binding studies the interaction of [3H][D-Ala2,D-Leu5]enkephalin (DADL) with its receptor was an extremely slow process. Rates of association and dissociation were determined and a pseudo-first order rate constant for association calculated to be 5.68 x 10(5) l/mol . s at 25 degrees C. The rate of dissociation (t 1/2 = 70 min) was accelerated by GTP and changed from linear to biphasic. The kinetically derived equilibrium dissociation constant (0.29 nM) was considerably lower than the KD obtained from Scatchard and Hill plots (1.24 nM). Measurement of DADL association as a function of temperature yielded a linear Arrhenius plot. Finally, competition binding assays revealed that delta-specific agonists exhibited relatively high potency in displacing [3H]DADL from synaptic plasma membranes receptors whereas mu-specific agonists and antagonists were less effective. These results with bovine hippocampus may be explained by the formation of a slow-dissociating, high affinity agonist conformation of the delta opiate receptor which has been predicted for the system by the cyclic-allosteric and ternary complex models. PMID- 6290247 TI - Isolation of large numbers of fully viable human neutrophils: a preparative technique using percoll density gradient centrifugation. AB - Techniques were developed for the isolation of human neutrophils from large volumes of human blood. This preparative procedure avoids exposure to Ficoll/Hypaque and offers a low cost alternative to centrifugal elutriation. First, leukocyte-enriched suspensions were obtained by treating buffy coats with 2% dextran. The leukocyte suspensions were then fractionated on three steps Percoll density gradients. The gradients were designed to carry large numbers of cells while maintaining resolution of the cell peaks. As a result, at least 320 X 10(6) leukocytes could be processed on a single 12 ml gradient. Extensive testing of the procedure showed that the purity of the neutrophil band was 96.5 +/- 3.16% (n = 50). The band contained 82.4 +/- 14.6% (n = 39) of the neutrophils applied to the gradient. Viability assays (bacterial killing, chemotaxis) demonstrated that neutrophils isolated on Percoll gradients were just as active as those harvested by centrifugal elutriation. Recovery data are presented and compared to the results obtained by others. PMID- 6290248 TI - Inhibition by cholera toxin of clonal growth of murine granulocyte/macrophage progenitor cells in soft agar cultures. AB - Proliferation in vitro of the committed granulocyte-macrophage progenitor cell (CFUC) is inhibited by cholera toxin (CT) in a dose-dependent manner. This inhibition is reduced and counteracted by higher doses of colony stimulating factor (CSF), an obligatory growth stimulator of CFUC. Mixing of CT with its specific receptor, the monosialoganglioside, GM1, before exposure to bone marrow (BM) cells, blocks the toxin's effect, and a restoration of colony formation is achieved. A dose-dependent inhibition of CSF-induced colony formation is also observed in the presence of choleragenoid, the B subunit of the toxin which binds to the specific receptor on the cell surface, but is biologically inactive. Incubation of BM cells with CT prior to cloning in soft agar cultures supplemented with CSF inhibits clonal proliferation of CFUC, whereas a brief in vitro exposure of BM cells to CSF prior to CT protects the cells against subsequent CT inhibition. The ability of CT to inhibit the CSF-induced clonal proliferation of CFUC and the effectiveness of CSF in reducing and even counteracting CT inhibition suggests that CT, by binding to BM cells through the B subunit, might either directly or indirectly interfere with the stimulatory activity of CSF upon its target cells. PMID- 6290249 TI - Some aspects of the regulation of the bronchial tree in obstructive lung disease: an introduction. AB - Hyperreactivity of the bronchial tree is a common characteristic in chronic obstructive lung disease. The mechanism underlying this phenomenon may be a disturbed equilibrium between contracting and relaxing forces acting upon the effector organ, the bronchial smooth muscle. The most important bronchial obstructive forces result from an increased vagal activity and from mediator release from the mast cell. The adrenergic system may be considered as a counter regulating force inducing a bronchodilatation. This system was studied by challenge tests with histamine, acetylcholine, and propranolol. Special attention will be paid to the relationship between the propranolol response and allergic characteristics in asthmatic patients. PMID- 6290250 TI - The bronchial tree and the beta-adrenergic system. PMID- 6290251 TI - The role of the adrenergic system in allergy and bronchial hyperreactivity. AB - The adrenergic system may have an influence on the severity and activity of bronchial asthma. It has been suggested that there might be a relationship between the allergic process and the adrenergic system. Therefore this system was studied before and after an allergen challenge. Two parameters were measured: in vivo: propranolol threshold measurement to assess adrenergic involvement; in vitro: the beta-adrenergic response of peripheral lymphocytes as parameter for beta-receptor function. Seven allergic asthmatics who were not treated with beta stimulants or corticosteroids and seven normals inhaled house dust mite extract. On the days before and after the challenge, propranolol threshold measurements were performed. Six patients had a dual reaction after a house dust mite inhalation; one patient had an early response. The propranolol threshold changed from 1.32% (SEM 0.16) on the day before challenge to 0.86% (SEM 0.19) on the day after (0.02 less than P less than 0.025). In the same patients the maximal cAMP response of lymphocytes in the presence of 10(-6) mM isoproterenol changed from 339% (SEM 18.9) above basal level before the allergen challenge to 194% (SEM 24.6) after the challenge (0.001 less than P less than 0.005). The pre-challenge beta-response of lymphocytes of the asthmatic patients was comparable with that of normal lymphocytes (298% SEM 59.8). The results suggest that an allergen induced asthmatic attack may lead to an impaired beta-adrenergic function. PMID- 6290252 TI - Transplantation of tectal tissue in rats. III. Functional innervation of transplants by host afferents. AB - Fetal tectal tissue was transplanted adjacent to the superior colliculus of neonatal rats. In mature recipient animals, ultrastructural studies provided evidence of host cortical synapses on cells within the transplants. To determine whether these synapses were functionally effective in driving transplant cells, tungsten in glass microelectrodes were used to record from single units in the transplants and the host cerebral cortex was electrically stimulated. Of the 214 tested units which were shown histologically to be within the transplants, 25 (11.7%) were orthodromically excited from host cortex at latencies of 15 ms or less (mean 7.3 ms). A further four transplant units were activated at relatively long latencies (40-300 ms). The presence of a functional host retinal input was also tested by electrically stimulating the host optic nerves. No transplant units were activated from these sites, apparently because very few electrode penetrations passed through the localized zones of optic axon distribution. The possible functional interrelationships between transplant and host are discussed. PMID- 6290253 TI - Leishmania mexicana: conversion of dihydroorotate to orotate in amastigotes and promastigotes. PMID- 6290254 TI - Experimental tumors with features of malignant fibrous histiocytomas. Light microscopic and electron microscopic investigations on tumors produced by cell transplantation of an established fibrosarcoma cell line. AB - An established fibrosarcoma cell line (RFS) derived from a cadmium induced fibrosarcoma of rat and RFS-cells in different subcultures produced tumors in nude mice and baby rats which showed characteristics of fibrosarcoma, malignant fibrous histiocytoma and completely undifferentiated sarcoma. After thorough examination of cells in culture and in experimental tumors by light microscopic and electron microscopic methods that conclusion has been drawn that malignant fibrous histiocytoma can develop from fibrosarcoma. By this the assumption that malignant fibrous histiocytomas derive from tissue histiocytes is questioned. After the discussion of the general unspecifity of the storiform growth pattern and the cytology of malignant fibrous histiocytomas the idea is presented that many defined malignant soft tissue tumors inclusively fibrosarcomas may pass a phase which should morphologically be diagnosed as malignant fibrous histiocytoma. Conceding the practical value of this diagnosis we emphasize that the concept of the entity "malignant fibrous histiocytoma" should be critically reevaluated because of the probable heterogeneity of this tumor group. PMID- 6290255 TI - The dietary origin of the urinary lignan HPMF. PMID- 6290256 TI - Effect of 5-thio-D-glucose on blood glucose and glucose-6-phosphatase activity in mice. PMID- 6290257 TI - Effect of ricin, of its subunits and of modeccin on cAMP level in Yoshida ascites cells. PMID- 6290258 TI - Cellular basis of the photoresponse of an extraretinal photoreceptor. PMID- 6290259 TI - Synthetic corticoliberin needs arginine vasopressin for full corticotropin releasing activity. PMID- 6290260 TI - Epidermal growth factor (EGF) accelerates the maturation of fetal mouse intestinal mucosa in utero. PMID- 6290261 TI - Action of ACTH, cAMP and cytochalasin B on steroid production by Y-1 mouse adrenal tumor cells in culture. PMID- 6290263 TI - Effects of VIP on glucose and lactate metabolism in isolated rat liver cells. PMID- 6290262 TI - [5'-Nucleotidase as an "internal probe" in studying steroid hormone interaction with the plasma membranes of target cells]. AB - Changes in the activity of 5'-nucleotidase, the marker enzyme of plasma membranes, in membrane cells of the uterus and liver were examined upon action of tropic and nontropic steroid hormones. In case of tropic steroids, the curves of changes in the activity with rise in the hormone concentration are complicated and broken with a marked minimum at 10(-8) M. Upon the action of nontropic hormones there takes place a monotonous rise in the enzymatic activity. The characteristic differences as shown by the curves make it possible to apply 5' nucleotidase as endogenous probe during studies into hormone-membrane interactions. PMID- 6290264 TI - A translational inhibitor activated in rabbit reticulocyte lysates under high pO2. AB - An inhibitor of protein synthesis was activated under high oxygen partial pressure (pO2) in hemin-supplemented and glutathione disulfide-free lysates from rabbit reticulocytes. This inhibitor shared some common features with other translational inhibitors from rabbit reticulocytes; that is, hemin-controlled repressor, glutathione disulfide-activated inhibitor and high pressure-activated inhibitor. It caused biphasic kinetics of inhibition which could be potentiated by ATP. Its activation was prevented by cAMP or glucose 6-phosphate. The high pO2 inhibitor could be partially purified from post-ribosomal supernatant containing ribosomal salt wash by precipitation between 0-50% (NH4)2SO4-saturation, Sephadex G-100, and DEAE-cellulose chromatography. PMID- 6290265 TI - An Asp-Asn substitution in the proteolipid subunit of the ATP-synthase from Escherichia coli leads to a non-functional proton channel. PMID- 6290266 TI - Isolation of mitochondrial porin from Neurospora crassa. PMID- 6290267 TI - On the inhibition of the b-c1 segment on the mitochondrial respiratory chain by quinone analogues and hydroxyquinoline derivatives. PMID- 6290268 TI - The nature of DNA breakage by 4'-[(9-acridinyl)amino]methane-sulphone-m anisidide. PMID- 6290269 TI - Determination of the conformations of cyclic nucleotides bound to the N-terminal core of the cyclic AMP receptor protein of Escherichia coli by 1H-NMR. PMID- 6290270 TI - Poly(ADP-ribose) synthetase inhibitors enhance streptozotocin-induced killing of insulinoma cells by inhibiting the repair of DNA strand breaks. PMID- 6290271 TI - An EPR study of the production of superoxide radicals by neutrophil NADPH oxidase. PMID- 6290272 TI - Hemin regulates the expression of transferrin receptors in human hematopoietic cell lines. PMID- 6290273 TI - Structure at restriction endonuclease MboI cleavage sites protected by actinomycin D or distamycin A. AB - Restriction endonuclease MboI cleavage of DNA was inhibited by actinomycin D and distamycin A. The two inhibitors protected different subsets of the 8 cleavage sites in polyoma DNA. The cleavage reactions were analyzed both in the presence of minimal inhibitory concentrations of the compounds and at higher concentrations, allowing cleavage at only 1 site/DNA molecule. The experiments showed that cleavage sites most efficiently protected by actinomycin D had putative inhibitor binding sites at a distance of 1-2 base pairs from the MboI recognition sequence. Distamycin A, in contrast, apparently has to bind immediately adjacent to the MboI recognition sequence to protect from cleavage. PMID- 6290274 TI - Regulation of smooth muscle contractile proteins by calmodulin and cyclic AMP. AB - The various protein components of a reversible phosphorylating system regulating smooth muscle actomyosin Mg-ATPase activity have been purified. The enzyme catalyzing phosphorylation of smooth muscle myosin, myosin-kinase, requires Ca2+ and the Ca2+-binding protein calmodulin for activity and binds calmodulin in a ratio of 1 mol calmodulin to 1 mol of myosin kinase. Myosin kinase can be phosphorylated by the catalytic subunit of cyclic AMP (cAMP)-dependent protein kinase, and phosphorylation of myosin kinase that does not have calmodulin bound results in a marked decrease in the affinity of this enzyme for Ca2+-calmodulin. This effect is reversed when myosin kinase is dephosphorylated by a phosphatase purified from smooth muscle. When the various components of the smooth muscle myosin phosphorylating-dephosphorylating system are reconstituted, a positive correlation is found between the state of myosin phosphorylation and the actin activated Mg-ATPase activity of myosin. Unphosphorylated and dephosphorylated myosin cannot be activated by actin, but the phosphorylated and rephosphorylated myosin can be activated by actin. The same relationship between phosphorylation and enzymatic activity was found for a chymotryptic peptide of myosin, smooth muscle heavy meromyosin. The findings reported here suggest one mechanism by which Ca2+ and calmodulin may act to regulate smooth muscle contraction and how cAMP may modulate smooth muscle contractile activity. PMID- 6290276 TI - Pain associated with hysterosalpingography: Ethiodol versus Salpix media. PMID- 6290275 TI - Isolation and properties of plasma membrane from smooth muscle. AB - A generalized approach to obtain relatively pure fractions of plasma membrane from smooth muscle tissues for studying calcium transport is described. The use of various markers for cellular membranes to establish the purity of various fractions is critically considered. Plasma membranes from rat myometrium have been isolated in a purity estimated to be 95-99%. Plasma membrane purifications to 70-80% have been achieved from rat mesenteric arteries and veins, canine tracheal smooth muscle, rabbit intestinal muscle, rat vas deferens, rat fundus, and dog gastric corpus. The ATP-dependent transport of Ca is correlated with the distribution of plasma membrane markers. Ca gradient of greater than 1000-fold have been achieved. ATP-dependent active Ca transport by plasma membranes could sometimes be stimulated by oxalate or phosphate. Anion activation of Ca active transport is not a marker for endoplasmic reticulum. In some smooth muscles (e.g., rat vas deferens) ATP-dependent Ca uptake did not correlate exclusively with the distribution of plasma membrane markers. Instead, the correlation seemed to be with NADPH-cytochrome reductase EC 1.6.2.5 activity (putative endoplasmic reticulum marker) as well as with plasma membrane markers. In all smooth muscles, active Ca transport appears to be a property of the plasma membrane; in some it may also be a property of the endoplasmic reticulum. Mitochondria actively transport Ca, but in most systems studied to date, the Km for Ca2+ for this transport is higher than that for plasma membrane. Thus the plasma membrane may be the major physiological mechanism of active transport for Ca out of cytoplasm of smooth muscle cells. In two plasma membrane fractions (from rat myometrium and mesenteric arteries) it has been possible to demonstrate the existence of an Na Ca exchange system. Its contribution to lowering cytoplasmic Ca is unknown. PMID- 6290277 TI - [Excitation-contraction coupling in the smooth muscle cells of the portal vein as affected by noradrenaline]. AB - Verapamil (10(-5) M) blocked phasic and tonic components of potassium contracture of smooth muscle cells (SMC) of the rat portal vein but failed to block completely noradrenaline, induced contraction of these cells, neither did it suppress the NA-induced contraction of the portal vein SMC in the presence of potassium contracture. These data suggest that NA-induced contraction is activated by external Ca2+ ions entering the SMC through chemosensitive (NA) voltage-independent Ca channels of the membrane. The contraction blocked by verapamil is activated by external Ca2+ ions entering through fast voltage dependent inactivating Ca channels participating in AP generation, and slow voltage-dependent inactivating Ca channels opened by NA depolarization. The basal tone and contraction evoked by transmitters and physiologically active substances in the SMC of all blood vessels seem to be activated primarily by external Ca2+ entering the SMC via the above mentioned types of Ca channels. PMID- 6290278 TI - Ultrasonic detection of fetal ascites associated with polyhydramnios. AB - The combination of fetal ascites and polyhydramnios is an uncommon problem that can be detected in utero by sonography. A case is reported where the etiology was found to be generalized cytomegalovirus infection. The differential diagnosis, etiology and diagnostic procedures are discussed. A thorough sonographic study is essential in the diagnosis and evaluation of fetal ascites in utero. PMID- 6290279 TI - Long-interval masculinizing Krukenberg tumor of the ovary. PMID- 6290280 TI - Iron-dependent free radical damage to DNA and deoxyribose. Separation of TBA reactive intermediates. AB - 1. Iron-dependent free radical damage to DNA and deoxyribose results in the formation of thiobarbituric acid (TBA) reactive intermediates. 2. These intermediates have been compared chromatographically and spectrophotometrically after incubation with the enzymes xanthine oxidase and peroxidase. 3. Loss of TBA reactivity occurred in the bleomycin-iron(II) derived products incubated with xanthine oxidase and in a standard solution of sodium malondialdehyde incubated with peroxidase. PMID- 6290281 TI - The membrane relationship of microsomal nucleoside diphosphatase. AB - 1. The effects of some agents which alter the activity or sedimentability of microsomal nucleoside diphosphatase (NDPase) were studied. 2. Trypsin activated NDPase by stimulating endogenous lipid peroxidation. 3. Eighty percent of the NDPase in vesicles was protected from anaerobic proteolytic attack as shown by activity measurements and electrophoresis. 4. Toluene activated NDPase which remained particulate. 5. NDPase and other proteins could then be easily released from microsomes. 6. The nature of the NDPase-membrane interaction is discussed. PMID- 6290282 TI - Characterization of six nucleoside-nucleotide phosphotransferases from the chromatin of Morris hepatoma 9121 cells by physicochemical and biochemical techniques. AB - 1. Six nucleoside-nucleotide phosphotransferases A-F were isolated from chromatin of Morris Hepatoma 9121 cells and characterized. 2. The apparent mol wt are: 13,000 daltons for A, 35,000 or 70,000 for B and E, 240,000 for C, 180,000 for D and 300,000-400,000 for F. 3. The sedimentation values are: 2.5-3 S for A, 3 or 5 S for B and E, 11 S for C, 7-8 S for D and 14 S for F. 4. The activity of the six enzymes A-F was stimulated with 1-2.5 micrograms calf thymus DNA using thymidine/ATP as substrates. 5. In the presence of defined polydeoxynucleotides, each enzyme A-F showed a differential preference for its substrates. PMID- 6290283 TI - Phosphotransferase properties of human erythrocyte phosphoglycolate phosphatase. AB - 1. Human erythrocyte phosphoglycolate phosphatase (PGP) (EC 3.1.3.18) shows transferase properties. Using p-nitrophenylphosphate (p-NPP) as substrate, methanol, at a concentration of 4.9 M, was the most efficient phosphate acceptor tested (60% phosphate transfer). 2. The branched alcohols i-propanol and i butanol accept the phosphate better than the unbranched compounds. The acceptor potency is methanol greater than ethanol greater than i-propanol greater than n propanol greater than i-butanol greater than n-butanol. 3. The relative transferase activity could be demonstrated to be independent of substrate concentration, pH, and the inhibitory effect of NaF at 2 and 4 mM. 4. PGP shows no transferase activity towards glucose and fructose, and is even inhibited by 250 mM of lactose and lactic acid to 67 and 55%, respectively. 5. Using p nitrophenyl-[32P]-phosphate (p-NP[32P]P), the direct transfer of phosphate from donor to acceptor could be demonstrated. PMID- 6290284 TI - Purification, isolation and characterization of a phosphoglycolate phosphatase isoenzyme from human erythrocytes. AB - 1. Preparation, purification and characterization of a phosphoglycolate phosphatase (PGP) isoenzyme from human erythrocytes was achieved by DEAE Sepharose CL-6B chromatography and isoelectric focusing using carrier ampholytes, pH 4-6. 2. The isoenzyme has an isoelectric point of 5.00 +/- 0.05 and could be purified 33,000 fold to a specific activity of 32.7 U/mg of protein. It represents the PGP phenotype 1 consisting of a single isoenzyme. 3. The enzyme is composed of two subunits (mol. wt 35,000) which are identical and not connected by SS-bridges. 4. At 4 degrees C the isoenzyme is more stable in the pH range of 7-9 than at acid pH values. 5. Incubation at 30 and 40 degrees C for 4 hr does not affect the activity of the isoenzyme. 6. It has a Km-value of 0.28 mM for phosphoglycolate (PG) as substrate. PMID- 6290285 TI - Candida utilis NAD+ kinase: purification, properties and affinity gel studies. AB - 1. NAD+ kinase (ATP:NAD+ 2' phosphotransferase, EC 2.7.1.23) has been purified to apparent enzymic homogeneity on Blue Sepharose CL-6B. 2. The molecular weight of the active species is about 260,000 as determined by PAGE and gel chromatography. Protein staining (PAGE) revealed minor bands with molecular weight values of 40,000, 140,000 and 550,000. Subunit studies (SDS-PAGE) gave evidence of a single band of molecular weight approximately 32,000. 3. On the basis of the release patterns of this enzyme from several affinity gels, an elution diagram is proposed as a device to assess the contribution of any of the several displacing agents that can be used to manipulate the desorption of a (enzyme) ligate from an immobilized ligand. PMID- 6290286 TI - Candida utilis NAD+ kinase: kinetic and inhibition studies with ADP. AB - 1. Initial velocity and product inhibition studies using ADP were carried out on cytoplasmic NAD+ kinase (ATP:NAD 2' phosphotransferase, EC 2.7.1.23) purified from Candida utilis. Initial velocity studies were also carried out on a sample of chicken liver NAD+ kinase. 2. The data indicate both enzymes followed a sequential mechanism of reactant binding. 3. Product inhibition studies on C. utilis NAD+ kinase suggest the mechanism of NAD+, ATP addition is best described as rapid equilibrium random with multiple binding of ADP to the free enzyme and the E X ATP and E X NAD+ complexes. 4. The characteristics of this enzyme, prepared from several sources, are briefly summarized. PMID- 6290287 TI - A new neuroregulator: the vasoactive intestinal peptide or VIP. AB - VIP is a neuroregulator occurring in the central and peripheral nervous system which exhibits the function of neurotransmitter in the brain, neuroendocrine substance at the pituitary level, and neuroparacrine substance in peripheral organs. The structure and the specificity of the molecule as studied by antibody and receptor, and its location in brain and peripheral organs are summarized as well as its numerous biological effects. The method used to demonstrate the involvement of VIP in a physiological regulation is described and illustrated by two examples: the effect of VIP on gut epithelium and the neuroendocrine action of VIP in inducing prolactin release from pituitary cells. The consequence of this recent progress in the knowledge of VIP release and action in human physiology and disease is indicated. PMID- 6290288 TI - Calcium facilitates endogenous proteolysis of the EGF receptor-kinase. PMID- 6290289 TI - Glycosaminoglycans in bovine cumulus-oocyte complexes: morphology and chemistry. AB - Bovine cumulus-oocyte complexes from small (1-5 mm) follicles were cultured for 24 h in 0.25 ml minimum essential medium supplemented with 10% fetal bovine serum and 20 microCi [3H]glucosamine. Treatment groups consisted of supplementing the culture medium with no hormone (control), 0.5 IU/ml follicle-stimulating hormone (FSH) or 10 mM 8-Br-adenosine cyclic monophosphate (cAMP). After culture, the complexes were fixed for light and scanning electron microscopy. Electron photomicrographs revealed that complexes induced to expand with FSH or cAMP contained a copious glycosaminoglycan (GAG) matrix extending between and around the cumulus cells. Control complexes did not exhibit expansion or an extracellular matrix. The radiolabeled GAG material was isolated for chemical identification. Chemical analyses included: (1) electrophoresis of GAG material, (2) electrophoresis of GAG material after enzyme or nitrous acid treatment, (3) thin-layer chromatography of GAG hydrolysates. The results from electrophoresis showed that the radiolabeled GAG co- migrated with hyaluronic acid. The GAG material was resistant to chondroitinase ABC and nitrous acid degradation but was digested by hyaluronidase. Complexes treated with FSH and cAMP incorporated higher (P less than 0.1 and P less than 0.025 respectively) amounts of [3H]glucosamine into hyaluronic acid than control cultures. Thin-layer chromatography identified the primary amino sugar of the GAG to be glucosamine. These data collectively showed that the radioactive GAG produced by bovine cumulus-oocyte complexes was hyaluronic acid. PMID- 6290290 TI - Cyclic nucleotide phosphodiesterase in developing rat testis. Identification of somatic and germ-cell forms. AB - Several forms of phosphodiesterase are present in the male gonad, and their relative concentrations vary during testicular development. On the basis of kinetic analysis and chromatography on DEAE cellulose, 3 forms were separated: (a) a high-affinity cGMP phosphodiesterase, regulated by Ca2+ and calmodulin, similar to a form described in different tissues (peak I); (b) a high-affinity cAMP form insensitive to Ca2+, which cannot be readily compared with forms described elsewhere (peak II); and (c) a high-affinity cAMP phosphodiesterase, Ca2+- and calmodulin-insensitive, corresponding to the "hormone-regulated" form described in several systems (peak III). The elution pattern of peak I and calmodulin stimulation were dependent on free calcium concentration during cytosol preparation and chromatography. This datum and rechromatography in the presence or absence of excess calmodulin suggested that the enzyme complexes calmodulin in a Ca2+-dependent manner and is therefore activated. Moreover, whereas peak I was observed in all cell compartments of the testis, peak II was present in germ cells and peak III was found mainly in somatic cells. During development of the testis, a relevant enhancement in the ratio between cAMP and cGMP hydrolytic activity was observed together with an overall increase of phosphodiesterase activity, thus suggesting that the previously described increase in cAMP phosphodiesterase activity during testis maturation should be ascribed to forms present in both germ cells and somatic cells. PMID- 6290291 TI - Effects of vitamin D3 metabolites on production of prolactin and growth hormone in rat pituitary cells. PMID- 6290292 TI - Functional properties of porcine-thyroid follicles in suspension. AB - Porcine thyroid follicle cells were isolated (about 10(7) cells per gram of tissue) and cultured in small aggregates in agarose-coated culture dishes. The aggregates became arranged into follicle-like structures capable of iodide uptake and organification. In the presence of TSH (0.2 mU/ml), the aggregation of follicles was enhanced, and iodide uptake as well as TSH-stimulated organification of iodide was increased compared with that in the control. In culture, the active iodide metabolism was gradually lost over a 7-day period. This was not due to a disappearance of the TSH-adenylate cyclase system, since cAMP production was retained and stimulated by TSH (half-maximal effect at about 1 mU/ml). Acutely TSH stimulated iodide efflux and iodide organification (half maximal effect at about 20 microU/ml). The stimulatory effect on organification was transient: within an hour further organification proceeded as in the absence of hormone. The effects on efflux and organification were already maximal at low TSH concentrations, whereas cAMP production was stimulated with up to 50-fold higher TSH levels, i.e. the findings were typical of spare receptors. In the continued presence of epidermal growth factor, a potent mitogen for thyroid cells, the follicles increased in size and contained one single large lumen. Their capability to take up and organify iodide was reduced. PMID- 6290293 TI - Studies on pituitary follitropin. XI. Induction of hormonal antagonistic activity by chemical deglycosylation. AB - Chemically deglycosylated ovine follitropin (DG-FSH) retained its binding ability to bovine and rat testicular membrane fractions but was completely inactive in stimulating cyclic AMP accumulation in immature rat seminiferous tubular suspensions. The native hormone induced cyclic AMP accumulation in a dose dependent manner but no significant response was detectable at any dose of DG-FSH even in the presence of a potent phosphodiesterase inhibitor. By virtue of these different biological characteristics all preparations of DG-FSH exerted potent inhibitory action when added to the cells along with the native hormone. Complete inhibition of FSH action was observed at equivalent concentrations of the DG-FSH. It was also effective against hFSH. DG-FSH had no effect on hCG action in rat interstitial cells. The accumulation of cyclic AMP induced by a non-hormonal agent such as cholera toxin in the tubular suspensions was unaffected by the concomitant presence of DG-FSH indicating that its antagonistic activity is mediated through its binding to specific follitropin receptors. PMID- 6290294 TI - Studies on pituitary follitropin. XII. Enhanced thermal stability induced by chemical deglycosylation. AB - The receptor binding, immunological and biological activities of ovine follitropin were rapidly destroyed when aqueous solutions were kept in a boiling water bath. Similar treatment of the chemically deglycosylated follitropin (DG FSH) showed less drastic effects on its hormonal properties. The latter preparation was more stable to heat treatment. A 30-min heat-treated DG-FSH was as active as native follitropin in the receptor-binding assay. Heat-treated DG FSH solution still retained its capability to antagonize the action of native follitropin in the in vitro bioassay. The conformational features of deglycosylated follitropin required for receptor binding and immunological reactivity are preserved after heat treatment. PMID- 6290296 TI - Adenylate cyclase in Xenopus laevis oocytes: characterization of the progesterone sensitive, membrane-bound form. AB - Progesterone-induced reinitiation of meiosis in Xenopus laevis oocytes involves a decrease in cAMP level. In these cells, adenylate cyclase is compartmentalized, with 25-30% in the plasma membrane fraction P-10000 (sedimenting at 10000 X g) and greater than 50% in the cytosol. Soluble adenylate cyclase appears not to be regulated via a GTP-binding regulatory protein (G/F) and is insensitive to progesterone. In contrast, membrane-bound adenylate cyclase seems to be linked to G/F, since it is stimulated by sodium fluoride, guanyl-5'-imidodiphosphate (Gpp(NH)p) and by cholera toxin; it is also inhibited by progesterone. The steroid inhibition is displayed towards basal and stimulated activities. The extent of progesterone inhibition of basal activity is dependent on Mg2+ and Mn2+ concentrations. The hormonal effect is independent of GTP concentration and is observed even in the presence of the non-hydrolysable analogue of GTP, Gpp(NH)p. The progesterone effect is not mediated by adenosine. Exposure of the P-10000 fraction to 5'-deoxy-5'-S-isobutylthioadenosine (a methyltransferase inhibitor) increases adenylate cyclase activity. PMID- 6290295 TI - Norepinephrine stimulates testosterone aromatization and inhibits 5 alpha reduction via beta-adrenoceptors in rat pineal gland. AB - The possible modulatory role of the sympathetic nervous system on testosterone aromatization and 5 alpha reduction by rat pineal gland was examined in vitro. NE (10 microM) added to pineal organ cultures increased by 72% the conversion of [14C]testosterone into estradiol and depressed by 39 and 53% that into the 5 alpha-reduced metabolites 5 alpha-dihydrotestosterone (5-DHT) and 5 alpha androstanediol (5 alpha-diol). Both effects of NE were negated by the addition of the beta-adrenoceptor antagonist propranolol but not by the alpha-adrenoceptor antagonist phentolamine. Dibutyryl cAMP (0.1 mM) mimicked the effect of NE on pineal [14C]testosterone metabolism; it also mimicked the NE-induced inhibition of [14C]progesterone reduction to 5 alpha-pregnanedione and 3 alpha-hydroxy-5 alpha-pregnan-20-one by rat pineal gland explants. At the end of the dark phase of the daily photoperiod, pineal aromatization of testosterone was significantly higher, and 5 alpha reduction lower, than in rats killed at noon. Pineal glands obtained from rats subjected to superior cervical ganglionectomy 12 h earlier exhibited increased conversion of [14C]testosterone into estradiol, and depressed synthesis of 5 alpha-reduced metabolites, as compared with their respective sham operated controls. 3 days after ganglionectomy a diminished testosterone aromatization was found. These results suggest that the increased release of NE from pineal sympathetic nerve endings stimulates testosterone aromatization and inhibits 5 alpha reduction via a beta-adrenoceptor. PMID- 6290297 TI - Archenteron formation induced by ascorbate and alpha-ketoglutarate in sea urchin embryos kept in SO2- 4 -free artificial seawater. PMID- 6290298 TI - Phenotypic dissections of the Blastocladiella emersonii zoospore's developmental choice. PMID- 6290299 TI - Evidence that intracellular cGMP is involved in regulating the extracellular cAMP phosphodiesterase and its specific inhibitor in Dictyostelium discoideum. PMID- 6290300 TI - Factors regulating prostaglandin uptake by rat small intestine. AB - The uptake of prostaglandin E1 by rat intestine mucosal strips was investigated. The results indicate that: (1) PGE1 did not accumulate in the epithelial cells of rat intestine. The intracellular to extracellular distribution ratio did not reach unity in spite of the extended periods of incubation. (2) A decrease in the sodium gradient across the brush-border membrane of intestinal cells significantly inhibited PGE1 uptake by the mucosal strips. (3) PGE1 uptake increases with increasing concentration of potassium in the incubation medium with a tendency for saturation at high K+ levels. (4) Calcium and magnesium ions had no effect on the steady-state uptake of PGE1 by the epithelial cells of rat intestine. (5) Addition of sodium carbonate or phosphate to the incubation medium significantly decreased PGE1 uptake by the intestinal cells. (6) Further studies of PGE1 uptake reveal a temperature-dependent process with a Q10 of 2.8. PMID- 6290301 TI - In vitro inhibition of pancreatic enzyme activities by dietary fiber. AB - Trypsin, amylase, lipase and phospholipase activities were assayed in buffer solutions and in human duodenal juice after incubation with different types of dietary fiber. In buffer solutions, trypsin activity was slightly reduced and amylase activity heavily reduced by pectin of low methylic esterification (LM pectin). Lipase activity was markedly reduced by LM pectin and also moderately reduced by pectin of high methylic esterification (HM pectin). Phospholipase was hardly influenced at all by fiber. Activities of pancreatic enzymes in human duodenal juice were examined after in vitro incubation with pectins, guar gum, wheat bran and ispaghula. Ispaghula did not affect the enzymes except for lipase activity, which was moderately inhibited. The other fiber preparations examined reduced amylase activity by 35-100% at fiber concentrations of up to 1.5 g%, lipase by 40-95% and trypsin by 40-85%. LM pectin had the strongest inhibitory effect among the different fiber preparations studied. Phospholipase activity was only influenced by the pectins, which caused a 75% reduction. It is concluded that dietary fiber of different kinds has the capacity to inhibit pancreatic enzyme activities. This inhibitory effect is dependent on the type of fiber and differently affects the different enzymes. Further, the inhibition seems to be more pronounced when exerted in human duodenal juice than in conventional buffer systems. PMID- 6290302 TI - Islet cell function in gold thioglucose-induced obesity in mice. AB - Blood insulin, blood glucose and the biosynthesis and release of insulin have been studied in mice made obese with a single injection of gold thioglucose. In such mice, blood glucose levels were normal, though serum insulin rose in parallel with the development of obesity. When compared with controls, insulin secretion and synthesis were increased in isolated islets of Langerhans from obese mice, over a wide range of glucose concentrations. However, in obese animals, insulin biosynthesis was augmented above control levels at 2 mmol/l glucose, whilst the increase in insulin secretion accompanying obesity only became evident at glucose concentrations greater than 5 mmol/l. After 2 min incubation, cyclic AMP rose more in islets from obese mice than in controls, though cyclic AMP levels did not significantly differ in either group after 10 min incubation with glucose. Glucose oxidation was also increased in islets of Langerhans from obese mice. It seems possible that changes in glucose oxidation, as well as in cyclic AMP levels, contribute to the alteration in the B cell response in this type of obesity. PMID- 6290303 TI - Quantitative comparison of Epstein-Barr virus receptor expression on sIgM and sIgG cell lines and B-cell lymphoma biopsies. AB - Over 50 B-cell derived lines and B-cell lymphoma and leukemia biopsies were screened for expression of the Epstein-Barr virus (EBV) receptor. The 13 sIgM positive lines bound more than five times as much virus as the six IgG lines. Among the biopsies, the 17 sIgM, 11 sIgM and sIgD, and seven sIgG expressing biopsies were further divided according to expression of the C3 receptor. C3 receptor-positive biopsies, which expressed sIgM alone or along with sIgD, had the largest subpopulation of cells which expressed the EBV receptor (EBVR). C3 receptor-negative biopsies only expressed the EBVR on half as many cells as their C3 receptor-positive counterparts. However, the relative number of EBVR on individual EBVR-positive cells was independent of C3 receptor expression. Within the sIgG class, both C3 receptor-negative and positive cells expressed equally low levels of EBVR, both in terms of subpopulation and relative number of EBVR per positive cell. These results suggest that subpopulation expression of the EBV receptor is related to the C3 receptor but that relative number of receptors per cell is associated with sIg phenotype. PMID- 6290304 TI - [Clinical study of a new cephalosporin ceftriaxone (Ro 13-9904) in sepsis and other infectious forms of particular severity]. PMID- 6290305 TI - Electron spin resonance study on the metabolism of 2-naphthylamine and 1 naphthylamine in rat liver microsomes. AB - Incubation of 2-naphthylamine (2-NA) with rat liver microsomes fortified with NADPH resulted in the formation of two types of free radicals which were identified as being due to N-hydroxy-2-naphthylamine and 2-amino-1-naphthol. The former radical was shown to be oxidized further to 2-nitrosonaphthalene radical. Compared with the case of the constitutive microsomes, the amount of the free radicals increased progressively on using the microsomes from methylcholanthrene (MC)-induced or phenobarbital (PB)-induced rats. On the other hand, the amount of the free radical formed from 1-naphthylamine (1-NA) was much less than that from 2-NA, so that the radical structure was not identified unambiguously. The enzymes involved in this case were not induced by either MC or PB. Thus, from the viewpoint of free radical formation, the metabolic patterns of 2-NA and 1-NA are quite different and such different behavior might be correlated with the distinct difference in their carcinogenicities. From the inhibitory effects of carbon monoxide, SKF-525A, alpha-naphthoflavone and 1-(1-naphthyl)-2-thiourea on the free radical formation, it was concluded that both mixed function oxidases (cytochrome P-450s) and mixed function amine oxidase (dimethylaniline monooxygenase) are involved in the metabolism of 2-NA. PMID- 6290306 TI - Sex steroid receptors in diverse human tumors. AB - The estrogen receptor was assayed in 8 intracranial meningiomas, 14 lung cancers, 3 thymomas and 2 malignant lymphomas using the dextran-charcoal method. Progesterone and androgen receptors were also assayed in all the cases of lung cancer. The estrogen receptor assay was positive in 3 meningiomas, 2 lung cancers, and all thymomas and malignant lymphomas. The androgen receptor assay was positive in one case of lung cancer but the progesterone receptor was negative in all assayed cases. No cases showed multiple receptor positivity. An antiestrogen (tamoxifen) was administered to one case each of intracranial meningioma and lung cancer with positive estrogen receptor. The receptor content was decreased to an undetectable level in the former after 4 months of administration. This finding suggests the estrogen dependency of the tumor. The lung cancer patient, however, died 2 weeks later without any evidence of hormone dependency. PMID- 6290307 TI - Acute viral enteritis and exacerbations of inflammatory bowel disease. AB - In order to determine whether or not acute viral gastroenteritis predisposes to exacerbations of inflammatory bowel disease, patients with Crohn's disease and ulcerative colitis were observed longitudinally. Assessment of disease activity was correlated with evidence for viral infection by serology and stool antigen testing. Disease exacerbations were rarely associated with rotavirus, Norwalk agent, or adenovirus infection, primarily because these infection rates were very low. However, the few patients having these viral infections often had relapse symptoms. PMID- 6290308 TI - A solid-phase sandwich radioimmunoassay for Entamoeba histolytica proteins and the detection of circulating antigens in amoebiasis. AB - A sensitive and specific immunochemical assay for Entamoeba histolytica antigens would be a valuable tool for clinical diagnosis, to study the sequelae of amoebiasis, and to screen for the expression of amoebic proteins in recombinant bacterial clones. The major impediment toward developing such an assay is the cross-reactivity of anti-E. histolytica antisera with a wide range of mammalian serum proteins. A rabbit anti-E. histolytica antiserum was repeatedly passed over three bovine serum protein immunoadsorbents and affinity purified over an E. histolytica protein-Sepharose 4B matrix. The purified antibody was radiolabeled and formed the upper layer of two specific and sensitive sandwich radioimmunoassays for amoebic proteins. One assay used a rabbit antiamoebic antibody solid phase and the other a human antibody solid phase. The latter assay proved capable of detecting amoebic antigens in the polyethylene glycol precipitates of sera from 21 of 21 patients with amoebiasis (and none of 22 control subjects). PMID- 6290309 TI - Cell culture of rat gastric fundic mucosa. AB - The purpose of this study was to develop a primary cell culture system of rat gastric fundic epithelial cells. The cells, isolated enzymatically, were cultured in Coon's modified Ham's F-12 medium supplemented with 10% fetal bovine serum, 15 mM HEPES buffer, fibronectin, and antibiotics. The inoculated cells started to grow rapidly on day 1 (doubling time, 26 h). The cells reached confluency on day 3. On phase contrast microscopy, over 90% of cells possessed epithelial characteristics. Histochemical studies showed (a) 90% of the epithelial cells contained PAS positive granules, (b) 5% of the cells gave a strong reaction for succinic dehydrogenase activity (presumably parietal cells), and (c) immunohistochemical localization of pepsinogen was negative. Ultrastructurally, microvilluslike structures, junctional complexes, Golgi apparatus, mitochondria, rough-surfaced endoplasmic reticulum, and mucous granules were observed. Mitotic figures were clearly observed on Giemsa staining and the mitotic index was maximum on day 2. Autoradiographic and biochemical studies showed these cells possessed the capability to synthesize deoxyribonucleic acid and this ability was maximum on day 2. These cells were able to synthesize and to secrete glycoprotein and this function was significantly increased by 16,16-dimethyl prostaglandin E2. Cyclic adenosine monophosphate produced by the cultured cells was enhanced by addition of 16,16-dimethyl prostaglandin E2 (p less than 0.01). This in vitro system provides a valuable model for studies of cellular functions of gastric mucosa. PMID- 6290310 TI - Can we diagnose amebiasis? PMID- 6290311 TI - In vitro studies on the effects of octopamine on locust fat body. PMID- 6290312 TI - [Specificity of Tn9 insertion into the genome of bacteriophage lambda att80 depending on its preceding location]. AB - It was shown that the site of previous integration (the donor site) of Tn9 affects the specificity of its next integration into the target molecule--phage lambda att80 DNA. The transposon integration sites were mapped by restriction and heteroduplex analysis following Tn9 transposition from chromosomal sites of Escherichia coli K-12 differing in location and Tn9 stability. When transposed from chromosomal galT::IS1 gene, Tn9 inserted into the site with coordinates 44,5 +/- 2 kb of lambda att80; when transposed from chromosomal attTn9A site, the transposon inserted into the sites with coordinates 31 +/- 0,7 kb or 33,3 +/- 0,5 kb. In the course of transposition of Tn9 from chromosomal attTn9N site the transposon inserted into the lambda att80 site with coordinates 26,5 +/- 5 kb. In the latter case, the increase of Tn9 single-stranded loop and the appearance of two new HindIII cleavage sites were observed in heteroduplex experiments. The data were interpreted as indicating structural rearrangements of Tn9 or linked sequences in the course of transposition. PMID- 6290313 TI - [Location of the traS- and finO-type genes and of the oriT region of plasmid R6K]. AB - The regions of B6K DNA corresponding to oriT, finO and traS are mapped using a number of hybrid plasmids and deletion mutants pAS3::Tn5, pAS3::Tn7 and pAS3::Tn9 obtained in vitro after treatment with restriction endonucleases EcoRI and BamHI. FinO- and traS-like genes were mapped in the regions of 10 and 25,0-25,4 MD, respectively, oriT being situated in the region of 4,6-4,7 Md. PMID- 6290314 TI - [Method of selecting mutations for the plasmid genes of streptomycin resistance in Escherichia coli K-12 cells]. AB - Evidence is presented that streptomycin-dependent mutants of Escherichia coli K 12 are incompatible with the functional activity of streptomycin-resistance genes coding for aminoglycoside adenilyl transferase. On the basis of this phenomenon, a method for selection of streptomycin-sensitive mutations in these genes is proposed, and the possibility of the direct selection of hybrid DNA molecules carrying inserts inactivating streptomycin-resistance genes is discussed. The effectiveness of the method for isolation of streptomycin-sensitive R100.1 plasmid derivatives was demonstrated. PMID- 6290316 TI - Nucleotide sequence of the transcription initiation region of a rat ribosomal RNA gene. AB - We cloned the part of the rDNA containing the transcription initiation region, and determined the exact site of initiation of the 45S RNA transcription. The nucleotide sequence of the region surround the initiation site was determined. Comparison of the mouse and rat genes revealed extensive homology between the initiation regions of the two species. Notably, upstream the transcription initiation site had higher homology (77%) than downstream (51%), suggesting functional significance of this region. PMID- 6290315 TI - The nucleotide sequence of the promoter region of hisS, the structural gene for histidyl-tRNA synthetase. AB - A plasmid has been constructed which carries hisS, the structural gene for histidyl-RNA synthetase of E. coli, on a 1.6-kb fragment bounded by PvuII and BstEII sites. The DNA sequence of both ends of this fragment was determined. The amino-terminal sequence of histidyl-tRNA synthetase was also determined to locate the promoter proximal coding region and the frame in which it is read. Three promoters were identified by consensus criteria. The region surrounding these promoters contains extensive twofold symmetry. PMID- 6290317 TI - Physical mapping of human adenovirus type 7 DNA using a simple and sensitive method of terminal labeling. AB - The tyrosine-containing peptide covalently attached to each 5'-terminus of adenovirus type 7 (Ad 7; Greider) DNA was labeled with 125I. The 5'-labeled DNA was subjected to digestion with several restriction endonucleases and the size of the labeled terminal fragments was determined. Partial hydrolysis by these endonucleases generated a series of labeled fragments which were fused to the terminal fragments and could, therefore, be detected by autoradiography. From the sizes of the partial products the location of the cleavage sites of the enzymes on Ad7 DNA could be determined. The subgenomic DNA extracted from incomplete particles by protease treatment could also be labeled with 125I, since it was found to contain the tyrosine-containing peptide covalently attached to the preferentially packaged left end of the genome. PMID- 6290318 TI - Cloning and expression of a Streptomyces fradiae neomycin resistance gene in Escherichia coli. AB - A Streptomyces fradiae DNA sequence, which codes for a neomycin phosphotransferase, has been subcloned from the Streptomyces recombinant plasmid pIJ2 [a chimera between the Streptomyces plasmid SLP1.2 and chromosomal DNA containing a neomycin (Nm) resistance gene] into the BamHI restriction enzyme site of pHV14. Three different recombinant plasmids (pWHR1, pWHR2, pWHR3) have been isolated which transform Escherichia coli to Nm resistance. Southern transfer hybridization experiments show that the recombinant plasmids contain the cloned Streptomyces Nm resistance gene, and lysates of E. coli containing the recombinant plasmids were shown to have Nm phosphotransferase activity, demonstrating that a gene from Streptomyces can be expressed in E. coli. PMID- 6290320 TI - Somatic DNA rearrangement generates functional rat immunoglobulin kappa chain genes: the J kappa gene cluster is longer in rat than in mouse. AB - The kappa immunoglobulin (Ig) genes from rat kidney and from rat myeloma cells were cloned and analyzed. In kidney DNA one C kappa species is observed by Southern blotting and cloning in phage vectors; this gene most likely represents the embryonic configuration. In the IR52 myeloma DNA two C kappa species are observed: one in the same configuration seen in kidney and one which has undergone a rearrangement. This somatic rearrangement has brought the expressed V region to within 2.7 kb 5' of the C kappa coding region; the rearrangement site is within the J kappa cluster which we have mapped. The rat somatic Ig rearrangement, therefore, closely resembles that seen in mouse Ig genes. In the rat embryonic fragment two J kappa segments were mapped at 2 and 4.3 kb 5' from the C kappa coding region. Therefore, the rat J kappa cluster extends over about 2.3 kb, a region much longer than the 1.4 kb of the mouse and human J kappa clusters. In the region between C kappa and the expressed J kappa of IR52 myeloma DNA, and XbaI site present in the embryonic kappa gene has been lost. A somatic mutation has therefore occurred in the intervening sequence DNA approx. 0.7 kb 3' from the V/J recombination site. Southern blots of rat kidney DNA hybridized with different rat V kappa probes showed non-overlapping sets of bands which correspond to different subgroups, each composed of 8-10 closely related V kappa genes. PMID- 6290319 TI - Gene organization of the transforming region of adenovirus type 7 DNA. AB - The sequence of the leftmost 11% of the weakly oncogenic human adenovirus type 7 (Ad7) DNA has been determined. This part of the Ad7 viral genome encompasses early region E1 which has been shown to be involved in the process of cell transformation in vitro (Dijkema et al., 1979). From the nucleotide sequence and determined coordinates of the E1 mRNAs, we are able to predict the primary structure of the polypeptides encoded by the transforming region of Ad7. The organization of the E1 region of Ad7 and of other adenovirus serotypes (Bos et al. 1981) leads to the proposal of a novel mechanism for gene regulation at the translational level in which protein synthesis can initiate at either the first or the second AUG triplet available in mRNA. The differences between the large E1b-specific tumor antigens of adenovirus types 12, 7 and 5 may explain the differences in oncogenicity of these viruses. PMID- 6290321 TI - The relationship between region E1a and E1b of human adenoviruses in cell transformation. AB - Baby rat kidney (BRK) cells were transfected either with intact region E1 DNA of adenovirus type 5 (Ad5) or with mixtures of DNA fragments containing the separated E1a and E1b regions. The results showed that mixtures of regions E1a and E1b transform with a similar efficiency as intact region E1. DNA fragments containing region E1b alone have no detectable transforming activity in primary BRK cells nor in established rat cell lines. When region E1a and Ad5 was combined with region E1b and Ad12 complete transformation was also obtained. Characterization of the cell lines transformed by separated E1a and E1b regions have led to the following conclusions: (1) Expression of region E1b is not dependent on specific linkage to region E1a as it occurs in the intact E1 region. (2) Region E1b is normally expressed into the corresponding major adenovirus T antigens (65,000 and 19,000 Mr with region E1b of Ad5; 60,000 and 19,000 Mr with E1b or AD12). (3) Region E1b of Ad12 can be activated by region E1a of Ad5 indicating that the Ela regions of both serotypes are functionally similar in transformation. (4) Cell lines containing region E1b of Ad5 are weakly oncogenic in nude mice whereas cells containing E1b of Ad12 are highly oncogenic in nude mice, indicating that the degree of oncogenicity is determined by region E1b. PMID- 6290322 TI - The isolation and partial characterization of linked alpha A- and alpha D-globin genes from a duck DNA recombinant library. AB - Genomic DNA from an adult duck (Cairina moschata) was used to construct a library of cloned DNA fragments in the vector lambda Charon 4A. Screening of the DNA library resulted in the isolation of a recombinant, D alpha G-1, which carries both the adult duck alpha A- and alpha D-globin genes. The two globin genes are separated by approx. 2.2 kb of DNA, they are encoded by the same DNA strand and their orientation with respect to the direction of transcription is 5'- alpha D- alpha A-3'. Partial sequence analyses indicate that the two alpha-globin genes contain intervening sequences at positions homologous to those in chicken and mammalian alpha-globin genes. PMID- 6290323 TI - A common sequence in the inverted terminal repetitions of human and avian adenoviruses. AB - The termini of the avian chick embryo lethal orphan (CELO) virus DNA have been sequenced. The results revealed a 63-bp-long inverted terminal repetition (ITR) which shared the sequence ATAATA with all adenovirus termini, thus far analyzed. The CELO virus ITR differed from those of the mammalian adenoviruses in two major aspects: (i) it is not a perfect duplication; (ii) it begins with a 5'-guanylic acid residue instead of the cytidylic acid normally observed in adenoviruses. PMID- 6290324 TI - Variation in mouse major urinary protein (MUP) genes and the MUP gene products within and between inbred lines. AB - The mouse major urinary proteins (MUPs) and the unprocessed in vitro translation products of MUP mRNA were each resolved by isoelectric focusing (IEF). The urinary MUPs showed about 15 distinct components, and the unprocessed MUPs about 20. In each case wide variation was observed in the relative intensities of individual bands. A comparison of three inbred lines (C57BL, BALB/c and JU) showed inter-line variation in the patterns both of the urinary MUPs and of the unprocessed MUPs. A series of experiments was carried out with a cloned MUP cDNA probe. All three inbred lines contain the same number (about 20) of MUP genes per haploid genome. In Southern blot analysis of genomic DNA the MUP genes displayed complex patterns which we interpret as showing variation on a common basic MUP gene sequence. For each combination of restriction enzymes tested, one size of fragment carried more than half of the total label, and this fragment was always the same in the three inbred lines. Inter-line differences were observed in the patterns of some of the less reactive fragments. MUP mRNA consists of at least two distinct species with sizes of 1 and 1.2 kb, which reacted with the probe in a label ratio of about 0.5 to 1. In the three inbred lines this ratio was essentially the same. PMID- 6290325 TI - Variation between mouse major urinary protein genes isolated from a single inbred line. AB - We describe ten Charon 4A genomic DNA clones from BALB/c mice which include at least seven different major urinary protein (MUP) genes. We have established the orientation of all seven sequences, and have placed six of them in precise register by means of restriction site maps and Southern blot hybridization with cloned cDNA sequences. Four of the seven genomic sequences (family I sequences) form hybrids with six independent cDNA clones that have a high thermal stability and hybridize more strongly with mRNA from three inbred mouse lines. Hybrids between the remaining three genomic sequences and the cDNA clones have a lower thermal stability and hybridize less strongly with mRNA from the three inbred lines. Homologies between different cloned sequences extend over as much as 15 kb. No clone contains parts of two MUP genes, and no homology has been detected between the 3' flanking region of one MUP gene and the 5' flanking region of another. PMID- 6290326 TI - M13 vectors for selective cloning of sequences specifying initiation of DNA synthesis on single-stranded templates. AB - M13 cloning vectors have been developed for the selection of DNA sequences capable of directing initiation of DNA synthesis on single-stranded templates. These vectors are derived from viable M13 mutants containing large deletions in the region of the complementary strand origin. The deletion mutants are defective in the conversion of viral single strands to the duplex replicative form (SS leads to RF) both in vivo and in vitro, give a reduced phage yield and form turbid plaques. A receptor site for foreign single strand initiation determinants has been introduced into the mutants by the insertion of EcoRI linker sequences at the deletion sites. Specific cloned sequences from bacteriophage G4 RF and from Co1E1 DNA restore a clear plaque type and normal phage growth. Selection of clear-plaque isolates obtained by transfection with RF from one of these vectors, M13 delta E101, carrying inserted Co1E1 HaeIII fragments resulted in the selective cloning of one specific fragment, the HaeIII-E fragment. Insertion of either the H or L strand of the HaeIII-E fragment into the M13 delta E101 viral strand gives a clear plaque phenotype, indicating the presence of initiation determinants on both the H- and L-strands of the Co1E1 HaeIII-E fragment. These cloning vectors provide a new means for the functional dissection of replication origins and for the identification of DNA sequences that determine the enzymatic mechanism of discontinuous synthesis along the length of the bacterial chromosome. The ability to assess initiation capability on the basis of plaque morphology also provides a means for rapid genetic analysis of initiation determinants. PMID- 6290327 TI - Selective cloning of Co1E1 DNA initiation sequences using the cloning vector M13 delta E101. AB - DNA sequences required to direct single-strand to double-strand conversion by a phi X174-type mechanism have been detected in Co1E1 plasmid DNA. The sites responsible for this DNA strand initiation, named rriA and rriB, have been localized to the L strand of the HaeII-E fragment and to the H strand of the HaeII-C fragment of Co1E1. To define rriA and rriB more precisely, random fragments of Co1E1 DNA have been cloned into the single-stranded phage cloning vector M13 delta E101. This phage contains a viable deletion of the M13 complementary strand origin and makes small turbid plaques. Cloning of DNA initiation determinants into the unique EcoRI site of M13 delta E101 rescues the mutant phage and restores the ability to form clear plaques. DNA sequence analysis of the random, 100-400 bp inserts in clear plaque isolates has localized the rriA and rriB determinants within DNA fragments of about 100 bp. These initiation determinants promote assembly of a functional, multiprotein priming complex, "primosome", identical to that utilized in the in vitro conversion of phi X174 single-stranded DNA to the duplex replicative form. PMID- 6290328 TI - Specific protection of nucleotides in the lac operator from DMS methylation and DNase I nicking by crude bacterial cell extracts. AB - Crude bacterial cell extracts prepared from an Escherichia coli lacIq strain were shown to protect specific nucleotides in the lac operator from methylation by dimethyl sulfate (DMS) or digestion by DNase I, whereas no protection was observed using extracts prepared from a nearly isogenic lacI- strain. These experiments show that it is not necessary to use purified regulatory proteins in experiments designed to localize sequences on DNA which interact with proteins. Therefore, crude cell extracts should be useful in DNA "footprinting" experiments to define regions of DNA which bind to unknown regulatory proteins. PMID- 6290329 TI - Molecular cloning and amplification of the gene for thymidylate synthetase of E. coli. AB - The thyA gene of Escherichia coli, which directs the synthesis of the enzyme thymidylate synthetase, has been subcloned from a recombinant lambda phage (Hickson et al., 1982) into the multicopy plasmid pBR325 to give the plasmid pPE245. To identify the thyA gene product, the transposon Tn1000 was inserted into pPE245 and derivative plasmids isolated that were no longer able to complement thyA mutations. When proteins synthesised by these plasmids and by pPE245 were labelled and analysed on SDS-polyacrylamide gels a protein of 33000 Mr, presumably the thyA+ gene product was absent whenever the thyA gene was inactivated. On assaying cell extracts prepared from cells harbouring pPE245 for thymidylate synthetase, the level of this enzyme was found to be elevated by a factor of at least 25. PMID- 6290330 TI - Physical localisation and direction of transcription of the structural gene for Escherichia coli ribosomal protein S15. AB - The coding sequence for the Escherichia coli ribosomal protein S15 (rpsO) has been shown to lie immediately adjacent to the structural gene for polynucleotide phosphorylase (pnp). Based on DNA sequencing data, it is deduced that rpsO is transcribed counterclockwise with respect to the standard E. coli genetic map. PMID- 6290331 TI - Isolation and characterization of yeast ring chromosome III by a method applicable to other circular DNAs. AB - A method is described for the isolation and purification of covalently closed circular (ccc) DNA from yeast (Saccharomyces cerevisiae). Spheroplasts are lysed at pH 12.45 which denatures linear but not ccc DNA. Next, the lysate is taken through a gentle high-salt-phenol extraction to remove single-stranded DNA. The ccc DNA, recovered by ethanol precipitation, can be further studied by agarose gel electrophoresis, can be cut with restriction endonucleases and can be used to transform Escherichia coli. This method efficiently purifies large (approx. 190 kb) and small (approx. 1.5 kb, TRP1-RI Circle) circular DNAs and thus has general applicability for isolation and purification of plasmids from yeast. PMID- 6290332 TI - Construction of a broad host range cosmid cloning vector and its use in the genetic analysis of Rhizobium mutants. AB - We have constructed a cosmid derivative of the low copy-number broad host-range cloning vector pRK290 (Ditta et al., 1980) by inserting a 1.6-kb Bg/II fragment containing lambda cos into the unique Bg/II site in pRK290. The new vector, pLAFR1, is 21.6 kb long, confers tetracycline resistance, contains a unique EcoRI site, and can be mobilized into and stably replicates within many Gram-negative hosts. We constructed a clone bank of Rhizobium meliloti DNA in pLAFR1 using a partial EcoRI digest. The mean insert size was 23.1 kb. When the clone bank was mated (en masse) from Escherichia coli to various R. meliloti auxotrophic mutants, tetracycline-resistant (Tcr) transconjugants were obtained at frequencies ranging from 0.1 to 0.8, and among these, prototrophic colonies were obtained at frequencies ranging from 0.001 to 0.007. pLAFR1 cosmids were mobilized from R. meliloti prototrophic colonies into E. coli and then reintroduced into R. meliloti auxotrophs. In most cases, 100% of these latter Tcr transconjugants were prototrophic. PMID- 6290333 TI - Bacteriophage T4 as a generalized DNA-cloning vehicle. AB - We have designed a method for inserting foreign DNA segments into bacteriophage T4. A plasmid containing T4 DNA is opened within the T4 sequence and the foreign DNA is inserted in vitro. Recombination in vivo, between T4 and the doubly chimeric plasmid, results in insertion of the foreign DNA into the genome of viable T4 phage. We have demonstrated the method by inserting a 203-bp DNA fragment from the lactose operon of Escherichia coli, into the dispensable region of the rIIB gene of T4. With minor modifications, the method should make possible the cloning of very large DNAs into any one of a large number of sites on the T4 chromosome. PMID- 6290334 TI - Physical map of the nrdA-nrdB-ftsB-glpT region of the chromosomal DNA of Escherichia coli. AB - Seven pLC plasmids (pLC 3-46, 8-12, 8-24, 8-29, 14-12, 19-24 and 42-17) which complemented nrdA, nrdB, ftsB and/or glpT mutations of Escherichia coli were analyzed. A restriction map of each plasmid was constructed and restriction fragments were subcloned into pBR322. A physical map of approx. a 15 X 10(6) Mr segment of the chromosomal DNA was deduced from the overlapping region of the pLC plasmids. The pLC plasmids and newly constructed plasmids were examined for the ability to rescue the mutations. The complementation tests defined the location of the genes in the 15 X 10(6) Mr segment in the following order: nrdA-nrdB-ftsB glpT. Functional nrdAB and ftsB genes were located in the 3.1 X 10(6) Mr EcoRI PstI fragment. PMID- 6290335 TI - Effect of strong promoters on the cloning in Escherichia coli of DNA fragments from Streptococcus pneumoniae. AB - Attempts to clone wild-type DNA containing the malM gene of Streptococcus pneumoniae in plasmid pBR322 of Escherichia coli were unsuccessful. However, it was possible to clone a PstI fragment of DNA containing this gene in a plasmid of S. pneumoniae. Cells carrying the recombinant plasmid produced large amounts of the malM product, amylomaltase, and a fragment of the protein coded by the adjacent malX gene, apparently as a result of transcription in opposite directions from strong promoters located between the two genes in the plasmid insert. Under derepressed conditions these products represented 10% of the total protein. No transcription terminators appeared to be included within the cloned segment. The effect of various mutations in the segment on its ability to be cloned in pBR322 was examined. Of those tested, only a down promoter mutation that affected production of both the amylomaltase and the X-protein rendered the segment clonable in E. coli. Fragments of the S. pneumoniae vector, pMV158, which appear to lack strong promoters, were readily cloned in the pBR322-E. coli system. Although it is possible that large amounts of the X-fragment are toxic for E. coli, a more general explanation would be that excessive transcription of the pBR322 vector portion interferes with maintenance of the recombinant plasmid. PMID- 6290336 TI - Physical mapping of the genome and sequence analysis at the inverted terminal repetition of adenovirus type 4 DNA. AB - The genome of adenovirus type 4 (Ad4), the unique member of Ad group E, has been mapped with nine restriction endonucleases. Comparison of the occurrence of restriction endonuclease cleavage sites on Ad2, Ad7, Ad12 and Ad4 indicates that there is very little homology between these serotypes. Sequence analysis at the ITR of Ad4 showed that the "CAT" box which is present in all the ITRs of Ad's so far sequenced is absent in Ad4. The length of 116 bp for the ITR of Ad4 is also different from that of other Ad subgroups. PMID- 6290337 TI - Versatile low-copy-number plasmid vectors for cloning in Escherichia coli. AB - Small low-copy-number plasmid vectors were constructed by in vitro and in vivo recombinant DNA techniques. pLG338 and pLG339 are derived from pSC105, have a copy number of six to eight per chromosome, and carry genes conferring resistance to tetracycline and kanamycin. pLG338 (7.3 kb) has unique restriction endonuclease sites for BamHI, SalI, HincII, SmaI, XhoI, EcoRI and KpnI, the first five lying within a drug resistance gene. pLG339 (6.2 kb) lacks the KpnI site, but has unique SphI and PvuII sites. These versatile vectors should be useful for cloning many genes coding for membrane and regulatory proteins which cannot be cloned into high-copy-number plasmids. PMID- 6290338 TI - Synthesis of the nutL DNA segments and analysis of antitermination and termination functions in coliphage lambda. AB - The nut antiterminator sequence, when present between a promoter and a terminator, permits the N-mediated antitermination of transcription in phage lambda. The efficiency of nutL was determined by assaying the activity of gene galK placed on a plasmid downstream from the promoter, nutL and terminator modules. As a reference and an estimate of the plasmid copy number, we have used an improved and very reproducible assay for bla activity. Sequences consisting of the 17-bp nutL core flanked by two HindIII cohesive sites were synthesized by the phosphite coupling method, and cloned in proper orientation between the Pp promoter of pBR322 and lambda gene N followed by the tL1 terminator on a galK expression plasmid. The antitermination efficiencies for two synthetic 17-bp nutL sequences, one wild type and one point mutant at the base of the nutL stem, are similar but substantially reduced in comparison with the native 25-bp nutL sequence cloned at the same site in the otherwise identical galK-expression plasmid. Multiple tandem insertions of the synthetic 17-bp nutL segment successively increase antitermination efficiency, but also to levels below those of comparable plasmids carrying multiple copies of the native 25-bp nutL sequence. Thus, several specific base pairs in the flanking sequences appear to be important for the efficient nut function. In an inverted orientation the 17-bp nutL sequence has lost its antitermination function. It also lost the termination activity exhibited by inversion of the longer 25-bp and 74-bp native nutL sequences. PMID- 6290339 TI - Human cytomegalovirus DNA: BamHI, EcoRI and PstI restriction endonuclease cleavage maps. AB - The cloned HindIII fragments of human cytomegalovirus (HCMV) strain AD169 DNA were mapped with respect to the BamHI, EcoRI and PstI restriction endonuclease cleavage sites. Composite restriction endonuclease cleavage maps for the entire virus genome were constructed using the previously established linkages between the HindIII fragments. PMID- 6290340 TI - [Radiobiological equivalent of benzo(a)pyrene carcinogenicity when it is inhaled]. PMID- 6290341 TI - [Preparation of an AqNaI sorbent for the extraction of carbon monoxide from the air]. PMID- 6290342 TI - [Hygienic evaluation of a textile dressing used in consumer services enterprises]. PMID- 6290343 TI - [Experience in identifying the source of benzo(a)pyrene contamination of the finished product in polyamine manufacture]. PMID- 6290344 TI - [Blastomogenic action of surface-active compounds, the components of synthetic detergents for the population]. PMID- 6290345 TI - [Hygienic evaluation of zeolites as filter material in practical water treatment]. PMID- 6290347 TI - [Clinical and pathogenetic characteristics of different forms of pneumoconiosis from exposure to quartz-containing dust]. PMID- 6290346 TI - [Content of natural radionuclides and benzo(a)pyrene in the environment in areas located next to the thermoelectric power plants operating on shale and mazut]. PMID- 6290348 TI - [Information from the Soviet Toxicology Center]. PMID- 6290349 TI - Bran works! PMID- 6290350 TI - Cytosolic estrogen and progestin receptors in human endometrium from different regions of the uterus. PMID- 6290351 TI - Identification and characterization of receptors for oxytocin in the myometrium of the pregnant ewe. AB - Receptors for oxytocin were identified in a particulate fraction from the myometrium of the pregnant ewe. Specific binding of [3H]-oxytocin was rapid, reversible, and saturable. It showed an absolute requirement for Mg2+ and the selectivity among oxytocin analogues expected for the receptor. On fractionation of the myometrium by differential and discontinuous gradient centrifugation, [3H] oxytocin binding showed a fractionation pattern which was similar to that for plasma membrane markers, but different from those for endoplasmic reticulum or mitochondrial markers. It is concluded that oxytocin receptors are located in the plasma membrane of the ewe myometrium. This subcellular fraction is a useful preparation for the study of receptor mechanisms and the regulation of myometrial contractility. PMID- 6290352 TI - Plasma adrenocorticotropic hormone, beta-lipotropin, and beta-endorphin in the human fetus at delivery. Correlation with birth weight and placental weight. PMID- 6290353 TI - Fibrinogen-bound sialic acid levels in the dysfibrinogenaemia of liver disease. AB - Fibrinogen-bound sialic acid levels were determined in 75 normal controls and 80 patients with liver disease. Patients with abnormal fibrin monomer polymerisation (FMP) had sialic acid levels significantly higher than controls or patients with normal FMP. Enzymatic removal of sialic acid from the abnormal fibrinogens corrected the abnormal FMP and thrombin-clotting times to the range of desialated controls. The accelerating effects of calcium ions, protamine sulphate and Polybrene were largely abolished by desialation, suggesting that these cations accelerate FMP by neutralising the negativity charged sialic acid residues. PMID- 6290354 TI - Lipid inclusions in oocytes and preimplantational embryos of different strains of mice. PMID- 6290355 TI - [Cephem antibiotics and alcohol metabolism: (1) Disulfiram-like reaction resulting from intravenous administration of cephem antibiotics]. AB - Fifty to 500 mg/kg doses of the cephem antibiotics were intravenously injected to male rats twice a day for 3 days. After the last injection, the rats were fasted for 17 hours and then orally administered 2 g/kg or 20% ethanol. The blood levels of ethanol and acetaldehyde (AcH) were determined by gas chromatography. Cefotiam, cefsulodin, and cefazolin did not affect the blood levels of ethanol and AcH as compared with those of the control. Cefmetazole, cefamandole, and cefoperazone did not change the blood ethanol level, but these antibiotics increased the blood AcH level dose-dependently. Cefamandole was especially able to sustain a high blood AcH level for over 8 hours. All of the antibodies which increased blood AcH levels contain the 1-methyl-1H tetrazole-5-thiol (TZ) group in their chemical structure. Intravenous injection of TZ caused a significant increase of the blood AcH level without influence on the blood ethanol level. 1 (2-Dimethyl-aminoethyl 1H-tetrazole-5-thiol (MTZ), the functional group which is contained in cefotiam, did not affect the blood levels of ethanol and AcH. These results suggested that the disulfiram-like reaction of cefmetazole, cefamandole, and cefoperazone results from an increase of the blood AcH level, and the 3 substituent group in in aminocephalosporanic acid, i.e., TZ, is an important factor for the reaction. PMID- 6290356 TI - [Effects of protizinic acid on the prostaglandins system and the production of oxygen radicals]. AB - Effects of protizinic acid (PRT) on prostaglandins (PG) and the production of oxygen radicals were compared with those of other non-steroidal anti-inflammatory agents. Oral administration of 30 mg/kg of PRT, indomethacin (IM), or ibuprofen (IB) significantly inhibited arachidonic acid-induced erythema in guinea pigs. Although 30 mg/kg of PRT significantly inhibited PGE2-induced erythema, IM and IB did not significantly inhibit it. PRT inhibited phospholipase A2 (PLA2) activity, and the IC50 value was 2.1 X 10-4 M. On the other hand, IM and IB exerted no effect on the PLA2 activity at 3 X 10-4 M. These results suggest that PRT possesses a broader pharmacological activity on the PG system than IM and IB. As for effects on the production of oxygen radicals, in order of relative inhibitory potency was PRT greater than metiazinic acid (MA) = IM greater than IB = phenylbutazone (PB) in the xanthine oxidase assay, PB great than IM greater than PRT greater than MA = IB in the rabbit neutrophil myeloperoxidase assay, and IM greater than PB greater than PRT greater than MA greater than IB in the guinea pig macrophage assay. In the rabbit neutrophil and aggregated IgG-bound micropore filter assay, the order was PRT greater than MA greater than PB greater than IM = IB. Thus, the inhibitory effects of PRT was verified in all experiments on the production of oxygen radicals in contrast to IB. In particular, it could be especially meaningful that PRT showed the most potent activity in the aggregated IgG-bound micropore filter assay which has been reported to be a good model for studying the pathogenesis of inflammatory diseases believed to be caused by immune complexes. PMID- 6290358 TI - [Effect of nitroglycerin on the heart]. AB - Nitroglycerin injection into the left coronary artery triggers an increase of ionized Ca concentration in coronary sinus blood. This coincides with a significant dilatation of the coronary artery and a decrease in left ventricular contractility. According to the results of the investigation it seems to be possible that nitroglycerin stimulates the Ca-ion efflux from the smooth muscle cells and the myocytes. PMID- 6290357 TI - Familial neuroaxonal dystrophy with principal lesions of nigro-pallido subthalamic degeneration. AB - Clinico-neuropathological studies were conducted on three patients of a family with parkinsonism. The clinical features of these three cases included juvenile parkinsonism with subcortical dementia. Neuropathological changes observed in common were the simple degeneration of the substantia nigra, globus pallidus and subthalamic nucleus, spheroid bodies in the reticular zone of the substantia nigra, intracytoplasmic deposits of an eosinophilic substance in the nerve cells of the locus caeruleus and a swelling of nerve cells of the thalamus and brainstem nuclei. In one of the cases, electron microscopy of the locus caeruleus demonstrated dense bodies scattering within meshes of tubules in the cytoplasm of nerve cells. All these findings led to the conclusion that the pathologic diagnosis was identified as familial neuroaxonal dystrophy with the principal lesions of nigro-pallido-subthalamic degeneration. PMID- 6290359 TI - [Progress in dermatology: new biochemical aspects]. AB - Recent biochemical advances have contributed to clarification of certain skin diseases and metabolic disturbances with predominantly cutaneous symptoms. This is illustrated by the various forms of porphyria. Today we differentiate four hepatic forms: acute intermittent porphyria, variegate porphyria, hereditary coproporphyria and porphyria cutanea tarda, and two erythropoietic forms: congenital erythropoietic porphyria and erythropoietic protoporphyria, all of which are due to an inborn enzymatic deficiency of the heme biosynthesis. From the different forms of ichthyosis, the X-recessive ichthyosis has an underlying enzymatic deficiency of the steroid sulfatase, which seems of significance in the disturbance of keratinization. In epidermolysis bullosa dystrophica type Hallopeau-Siemens an increased collagenase activity was detected. Inhibition of this enzyme by phenytoin results in improvement of the blistering in this genodermatosis. The etiology and pathogenesis of psoriasis are unclear despite extensive efforts. The recently detected deficiency of the arylhydrocarbon hydroxylase and its inducibility must be confirmed, additionally its significance in the pathogenesis of this disease is yet to be evaluated. PMID- 6290360 TI - [Therapeutic effect of lithium]. PMID- 6290361 TI - Ontogenesis of hepatic growth hormone receptors in the rat. AB - Detailed ontogenic studies of the binding of human (hGH) and bovine growth hormone (bGH) have been performed in liver preparations from male and female rats during the neonatal, weanling, pre- and post-pubertal periods. Specific binding of both hormones was readily detected at all ages, with no apparent interference due to occupancy by endogenous hormones. No sex difference in binding was observed prior to weaning (22 days) for hGH, which binds to both somatotrophic and lactogenic sites. However, after weaning a marked sex-related dissociation in the pattern of binding did occur, with female rats binding 3-4 times more hGH than in the pre-weaning period and male rats binding hGH to only half their pre weaning levels. A very similar pattern was seen for binding of bGH (which binds only to somatotrophic sites) except that in male rats, the post-weaning levels did not fall. Binding patterns for either hGH or bGH prior to weaning did not mirror the known age-related pattern of circulating rat GH levels, suggesting the absence of a definitive auto-regulation system for the GH-GH receptor system under normal circumstances in vivo. The possible role of the weaning process per se in the post-weaning changes of GH binding seen in male and female rats still requires elucidation. PMID- 6290362 TI - Late-onset 21-hydroxylase deficiency is an allelic variant of congenital adrenal hyperplasia characterized by attenuated clinical expression and different HLA haplotype associations. AB - Three families with late-onset 21-hydroxylase deficiency were studied. Homozygous females presented with symptoms of mild hyperandrogenism such as acne, hirsutism, oligomenorrhea and menometrorrhagia. A homozygous male was asymptomatic and had reached normal adult height. The diagnosis of 21-hydroxylase deficiency was based upon markedly elevated responses of plasma 17-hydroxyprogesterone during a short (30-min) ACTH infusion test. The propositi of two of the families were diagnosed despite long-standing glucocorticoid therapy and adrenal suppression by using a prolonged (48-hour) ACTH infusion. Heterozygotes of late-onset 21-hydroxylase deficiency had mildly elevated 17-hydroxy-progesterone responses to ACTH. Late onset 21-hydroxylase deficiency was inherited as an autosomal recessive trait with close linkage to the histocompatibility leukocyte antigens. The B14 haplotype was present in all affected members. One affected female had a daughter with classic, salt-losing 21-hydroxylase deficiency. Mixed heterozygosity of this patient for a classic and a late-onset 21-hydroxylase deficiency allele may have caused the classic phenotype in her daughter (homozygote for 2 classic alleles). PMID- 6290364 TI - The optimal length of hospitalization for psychiatric patients: a review of the literature. AB - In recent years, several controlled studies have evaluated the value of different lengths of psychiatric hospitalization and of alternatives to hospitalization. The author reviews such studies, noting that most findings suggest that longer stay does not decrease subsequent hospitalization, and does not clearly improve social adjustment or diminish psychopathology. Longterm hospitalization may increase the patient's commitment to continued psychiatric care, but short stay with optimal aftercare planning may be just as beneficial. Long-term hospitalization is necessary clinically for some patients, but the evidence is consistent and convincing in indicating that hospitalization should be kept as short as feasible. PMID- 6290363 TI - Synthesis of immunogenic C-6 derivatives of 2-methoxyestrone and 2 methoxyestradiol-17 beta and characterization of the corresponding antisera. AB - The synthesis, purification and structural confirmation of the 6 (carboxymethoxyimino) derivatives of 2-methoxyestrone and 2-methoxyestradiol-17 beta are described. These derivatives were coupled to bovine serum albumin by the mixed anhydride method, and rabbits were immunized with the product. The resulting antisera showed high affinity and specificity for 2-methoxyestrone and 2-methoxyestradiol-17 beta, respectively, with low cross reactivities to structurally related estrogens. PMID- 6290365 TI - Rapidly growing lesions on the abdominal skin. PMID- 6290366 TI - Tissue mast cells in immediate hypersensitivity. PMID- 6290367 TI - Kaposi's sarcoma: an unexpected story. PMID- 6290369 TI - Chemodectoma involving the cavernous sinus and semilunar ganglion. AB - This paper reports a case of paraganglioma involving the cavernous sinus and semilunar ganglion. The origin of the tumor cells, from paraganglia in this region, is also discussed. Recognition of paraganglia in this area may aid the clinician and pathologist in the differential diagnosis of tumors in this region. PMID- 6290368 TI - Adenoid cystic carcinoma: a comparative pathologic study of tumors in salivary gland, breast, lung, and cervix. AB - Histologic, histochemical, and ultrastructural features of eight adenoid cystic carcinomas arising at diverse sites were compared in order to determine diagnostic values and to investigate histogenetic mechanisms. These tumors originated in the salivary glands, breast, uterine cervix, and tracheobronchial tree. By light microscopy each tumor was seen to have morphologic features of adenoid cystic carcinoma, yet only five of the eight cases showed differential staining for the two mucin types, stromal and epithelial, which are reportedly present in these tumors. In contrast, every case showed a set of fine structural features which, in aggregate, are specific for adenoid cystic carcinoma. These features include pseudocysts, intercellular spaces, basal lamina, and true glandular lumens. The most prominent feature is the pseudocyst, which mimics a glandular lumen by light microscopy but is actually a rounded extracellular space containing basal lamina. Ultrastructurally, the variation in composition of the extracellular compartments, including pseudocysts and true lumens, appears to explain the lack of uniformity in the histochemical staining. The tumors also contained cytoplasmic microfilaments in parallel bundles, consistent with myofilaments. The presence of these filaments combined with basal lamina suggests myoepithelial differentiation, yet it is not known whether these tumors truly originate from myoepithelium or show differentiation toward myoepithelium as a part of the neoplastic process. Regardless of their histogenesis, this study shows that true adenoid cystic carcinomas do arise in different organs. Knowledge of the specific ultrastructural features of adenoid cystic carcinomas can be useful in classifying these tumors in some cases. PMID- 6290370 TI - Kaposi's sarcoma following chemotherapy for testicular cancer in a homosexual man: demonstration of cytomegalovirus RNA in sarcoma cells. AB - A 35-year-old white Jewish homosexual man who had undergone surgery and chemotherapy for an embryonal carcinoma of the testis subsequently developed Kaposi's sarcoma. The neoplasm involved the skin as well as visceral tissues. Tissue derived from a biopsy specimen of one of the skin lesions was used in the in situ hybridization technique for the detection of genetic material. Cytomegalovirus messenger RNA was identified in the neoplastic Kaposi cells in the skin. The significance of this finding is discussed. PMID- 6290371 TI - Granular cell variant of neurofibromatosis: ultrastructure of benign and malignant tumors. AB - A case of neurofibromatosis with a metastasizing malignant tumor is presented. The benign lesions were identified by light microscopy as neurofibromas containing granular cells typical of granular cell tumors. The malignant tumor was classified as a malignant granular cell tumor. Ultrastructural studies demonstrated common histogenetic and subcellular features of the benign and malignant tissues. The case is the first reported in which granular cells are seen in the lesions. The typical granular cell morphologic features of the malignant lesion support the concept of Schwann cell derivation of this group of tumors. PMID- 6290373 TI - Purine metabolism in fibroblasts of patients with Duchenne's muscular dystrophy. PMID- 6290372 TI - Complete moles have paternal chromosomes but maternal mitochondrial DNA. PMID- 6290374 TI - Anchorage independent growth of SV40 transformed human epithelial cells from amniotic fluids: differences within and among cell donors. AB - Cells from seven individuals were cultured separately for the induction of morphological transformation by SV40. Sixty-three transformed colonies were tested for anchorage independent growth in soft agar at various passage levels. Colony formation was consistent for all clones of respective cell donors. Four donors yielded clones that grew in soft agar in the first passage. Clones from three donors were similar to controls and formed no colonies. The size of the agar colonies was constant in the early passages. Size differences were observed in later passages and for negative clones that gained anchorage independence during time in culture. The early passage positive type of anchorage independence is expressed concomitantly with morphological transformation. Considering that the clonal isolates were genetically homogeneous within cell donors and heterogeneous among cell donors, it is concluded that the phenotype of anchorage independence is determined by at least two genetic mechanisms; namely, the genotype of the cell donor (the hereditary type) or by culturally derived new genetic variability, or both. Family history on cancer incidence showed that one grandparent for each of the four positive donors for the hereditary type of anchorage independence had cancer, whereas the grandparents of the three negative donors were asymptomatic. The incidence of cancer did not appear to be age related. Chromosome analyses of two morphologically transformed colonies from each of the cell donors by the in situ technique, showed diploid and tetraploid cells and a small number of cells with rearrangements. It is concluded as previously that the progenitor transformed cell to the colony of cells is normal diploid. PMID- 6290376 TI - Strain-specific differences of the major structural proteins of murine mammary tumour virus. PMID- 6290377 TI - Structure of DNA--binding to hydroxyapatite as a probe. PMID- 6290375 TI - A test for hormonal responsiveness in a mammary epithelial cell line, NMuMg. AB - The NMuMG cell line derived from normal mouse mammary epithelial cells was tested for responsiveness to hormones. The hormones studied included insulin, glucocorticoids (cortisol and dexamethasone), and prolactin. In addition to membrane bound insulin receptors and prolactin receptors, the cells had 2 X 10(4) cytoplasmic glucocorticoid receptors per cell. Morphological changes were observed in response to hormones. Clusters of cells appeared with greatly increased diameter, and the number of cells per plate was reduced. The rate of DNA synthesis, corrected by cell number, indicates that cell division, and hence cell turnover, was increased by the combination of all three hormones. Insulin greatly enhanced protein synthesis, but glucocorticoid and prolactin did not further increase the rate. The combination of three hormones produced a change in the synthesis of histones, consistent with the increase in cell turnover. There were substantial responses of enzyme activities to hormonal treatment of the cells. Insulin by itself induced a doubling of the activity of glyceraldehyde phosphate dehydrogenase and perhaps a modest increase in NADH-cytochrome c reductase. Lactose synthetase activity showed a three- to fourfold induction of both A and B subunits of the enzyme when the cells were treated with insulin, glucocorticoid, and prolactin, and the effect of the latter two hormones was shown to be additional to that of insulin. PMID- 6290378 TI - [The pathogenetic importance of peanut lectin binding sites in infectious, toxic and neoplastic processes]. AB - Labelled lectins are generally used for identification of mono-, oligo- and polysaccharides in histochemistry. The aim of this work is to demonstrate in addition the importance of the labelled (FITC and peroxidase) lectin from peanut (peanut agglutinin = PNA) with regard to the pathogenesis of some infectious and neoplastic diseases. Thereby, PNA-binding sites have been shown as differentiation marker in embryonic kidneys and in dysontogenetic. PMID- 6290379 TI - Development of acute autoimmune encephalomyelitis in mice: factors regulating the effector phase of the disease. AB - The development of acute experimental autoimmune encephalomyelitis (EAE) in mice is potentiated by the use of Bordetella pertussis vaccine as an adjuvant. Histamine sensitizing factor (HSF) extracted from B. pertussis is the active adjuvant agent and causes a mild increase in cerebrovascular permeability. During the development of EAE, there is an additional increase in vascular permeability of the brain and spinal cord. The adjuvant action of B. pertussis HSF does not appear to mimic a generalized beta-adrenergic blockade, since the course of EAE is not potentiated by adrenalectomy. The cerebrovascular permeability changes observed in EAE are probably mediated by vasoactive amines, since the expression of EAE can be blocked by vasoactive amine antagonists. PMID- 6290380 TI - Cyclic AMP reduces and cyclic GMP increases the traffic of lymphocytes through peripheral lymph nodes of sheep in vivo. AB - Acute and chronic infusions of a lipophilic analogue of cyclic AP (dibutyryl cyclic AMP) into cannulated afferent lymphatics of popliteal lymph nodes of sheep have reduced the output of both small and blast lymphocytes into efferent lymph and have tended to reduce the volume of efferent lymph flow (collected by chronic cannulation). Similar infusions of a cyclic GMP analogue (8-bromo cyclic GMP) have increased the output of both small and blast lymphocytes into efferent lymph and the volume of efferent lymph flow. It is suggested that cyclic nucleotide metabolism of lymphocytes and of vascular endothelium may play an important role in the regulation of lymphocyte traffic through peripheral lymph nodes in vivo and that secondary alterations in cellular cyclic nucleotide levels may be a common pathway by which the many and varied agents arriving in afferent lymph may affect lymphocyte traffic. PMID- 6290382 TI - Evidence for the presence of specific receptors for inhibin in human prostate. PMID- 6290381 TI - T-cell regulation of pokeweed-mitogen-induced polyclonal immunoglobulin production in mice. III. Role of Lyt 1+2- non-helper T cells in the suppression. AB - A large number of cells which can replicate vesicular stomatitis virus (VSV) were generated in pokeweed mitogen (PWM)-stimulated cultures of normal mouse spleen cells. The optimal dose of PWM for the generation of VSV-replicating cells was within the range 25 and 50 micrograms/ml, which had previously been shown to be optimal for the induction of suppressor cells. The VSV-replicating cells in this system were shown to be Thy 1+ and Lyt 1+2-. These cells, however, were unlikely to be helper T cells, but may be a subset of T cells involved in suppression. Thus, inoculation of VSV into a PWM-stimulated culture of spleen cells resulted in marked augmentation of polyclonal immunoglobulin production. Further, it was shown that the suppressive activity of T cells which had been precultured with a high dose of PWM was abolished by the infection with this virus. PMID- 6290383 TI - Binding characteristics of inhibin to rat pituitary plasma membrane. PMID- 6290384 TI - Physiochemical properties of polymorphonuclear leukocyte surface structures associated with the f-Met-Leu-Phe receptor. AB - Surface property changes of polymorphonuclear leukocytes (PMNL) as a result of development of functional receptors to f-Met-Leu-Phe and as a result of f-Met-Leu Phe binding have been studied by aqueous biphasic partitioning of the cells in systems of dextran and polyethylene glycol (PEG) with part of the PEG exchanged for positively charged or hydrophobic PEG. The Phe was associated with increased exposure of hydrophobic surface structures as well as of negatively charged groups on the PMNL surface. Binding of f-Met-Leu-Phe to the PMNL surface receptors caused hiding of hydrophobic and charged structures, indicating that these properties are of importance in the interaction between PMNL and the chemotactic peptide. PMID- 6290385 TI - Influence of dietary omega-3 fatty acids on granulocyte function. AB - Manipulation of dietary fatty acid content has been shown to influence platelet aggregation responses. Because granulocytes and platelets interact in a variety of biologic systems, we wondered whether a similar effect might be observed on granulocytes. Granulocyte function was therefore studied in three donors prior to and after three weeks upon a diet supplemented with large amounts of eicosapentaenoic acid. The previously reported attenuation of platelet aggregation was observed, but no effect was seen on granulocyte aggregation, chemotaxis, or superoxide production. Although several other explanations are possible, we suggest that the most likely explanation for this dichotomy is that granulocyte aggregation and chemotaxis are not centrally dependent upon production of thromboxane A. PMID- 6290386 TI - Influence of lithium and fluoride on degranulation from human neutrophils in vitro. AB - We have demonstrated that degranulation from normal human neutrophils in whole blood was stimulated by low concentrations of lithium salts and was produced by noncytolytic means. Significant amounts of beta-glucuronidase could be released from the primary granules, in addition to vitamin B12- binding protein from the secondary granules, by 10 mM lithium. Release was almost totally inhibited by 1 mM fluoride, under the same conditions. There was no release of lactate dehydrogenase and no loss of viability of cells incubated in either lithium or fluoride at the concentrations used. Lithium was also observed to have no effect on reactive oxygen production by phagocytic stimulation of isolated neutrophils. In addition, lithium and fluoride were shown to manipulate the intracellular levels of cAMP. The results demonstrated a direct effect of lithium on release of inflammatory mediators from neutrophils in vitro. The methods used also provide a simple and effective test to study an important function of neutrophil activity and can be used to evaluate degranulation in a number of clinical conditions. PMID- 6290387 TI - Infection of marmosets with parainfluenza virus types 1 and 3. AB - Infection of wild marmosets (Saguinus mystax) with strains of parainfluenza virus types 1 and 3 resulted in acute respiratory infection. Virus replication in the upper respiratory tract was of a degree similar to that seen in children acutely infected with parainfluenza viruses. Serum antibody developed with both virus types; however, local secretory antibody was not detectable. The infection was transmissible to susceptible animals up to 3 days inoculation of the primary animal. PMID- 6290388 TI - Role of macrophages in innate and acquired host resistance to experimental scrub typhus infection of inbred mice. AB - Mechanisms of innate resistance to infection with the Gilliam strain of Rickettsia tsutsugamushi were examined using congenic strains of mice resistant (C3H/RV) or susceptible (C3H/He) to intraperitoneal infection. Both strains of mice were resistant to infection with 1,000 50% mouse lethal doses of rickettsiae if given intravenously. In both systems rickettsial replication occurred after intravenous infection, as evidenced by an increase in rickettsial numbers in the spleens of infected animals, followed by a decrease in rickettsiae to low levels by day 14 postinfection. Administration of the antimacrophage agents silica and carrageenan to C3H/He mice intravenously rendered these animals susceptible to lethal infection. Neither irradiation nor silica given individually rendered C3H/RV mice susceptible to intravenous infection. However, if silica and irradiation were given together, a lethal infection occurred after intravenous infection. C3H/RV mice became susceptible to lethal infection after sublethal doses of irradiation only if they were infected intraperitoneally. Administration of silica or carrageenan had no effect on the outcome of intraperitoneal infection of these mice with Gilliam rickettsiae. These data suggest that both strains of mice share innate resistance mechanisms to intravenous infection that consist of fixed macrophages. Resistance of C3H/RV mice to intraperitoneal infection, in contrast, apparently was dependent only on an irradiation-sensitive process. PMID- 6290389 TI - Resistance to infections in mice with defects in the activities of mononuclear phagocytes and natural killer cells: effects of immunomodulators in beige mice and 89Sr-treated mice. AB - Beige mice, which are a homolog of the Chediak-Higashi syndrome, and mice treated with 89Sr to destroy the bone marrow provide animal models of defects in mononuclear phagocyte and natural killer cell functions. The innate resistance of these mice to viruses such as herpes simplex and encephalomyocarditis viruses, however, is normal. Moreover, treatment of the mice with immunomodulators such as Propionibacterium acnes (formerly designated Corynebacterium parvum) and pyran produced a significant increase in resistance to encephalomyocarditis virus. The antiviral effect of P. acnes in 89Sr-treated mice was exhibited during marked monocytopenia and without evidence for an inflammatory influx of macrophages into the peritoneal cavity. Treatment with P. acnes was also effective in increasing the resistance of beige mice to infection with Listeria monocytogenes. Thus, immunomodulators can be effective in mice that exhibit impaired macrophage and natural killer cell functions. PMID- 6290390 TI - Protection against lethal challenge of BALB/c mice by passive transfer of monoclonal antibodies to five glycoproteins of herpes simplex virus type 2. AB - Monoclonal antibodies secreted by six hybridomas and recognizing antigenic sites on glycoproteins gC, gAB, gD, gE, and gF of herpes simplex virus type 2 were examined for their ability to protect BALB/c mice from lethal infection by the virus. Administration of monoclonal antibodies to individual glycoproteins intraperitoneally 3 h before footpad challenge with 10 times the 50% lethal dose of virus protected between 35 and 75% of the mice, except for one of two monoclonal antibodies recognizing antigens on gC. The antibodies did not neutralize virus in vitro and protected A/J mice deficient in the fifth component of complement as efficiently as complement-sufficient BALB/c mice. A good correlation was found between protection and titers of monoclonal antibodies assessed by antibody-dependent cell-mediated cytolysis. The results indicate that any of the glycoproteins can serve as antigens for a protective immune response. In addition, the data are compatible with protection being mediated by an antibody-dependent cell-mediated cytolysis mechanism. PMID- 6290391 TI - Role of Ia antigen expression and secretory function of accessory cells in the induction of cytotoxic T lymphocyte responses against herpes simplex virus. AB - Splenocytes from mice which have been primed in vivo with herpes simplex virus type 1 (HSV-1) can be restimulated in vitro with infectious or UV-inactivated HSV 1 to generate HSV-specific, H-2-restricted cytotoxic T lymphocytes (CTL). HSV primed splenocytes which have been depleted of adherent cells by sequential incubation on plastic, nylon wool, and Sephadex G-10 are not able to respond with a CTL response when restimulated in vitro. A variety of Ia-positive and Ia negative (Ia(+) and Ia(-)) cell populations were assessed for their ability to supply accessory cell functions to spleen cells which had been exhaustively depleted of adherent cells, as measured by the restoration of a CTL response to HSV-1. Of these, only those populations which included Ia(+) cells were capable of providing accessory cell function. The ineffective populations were devoid of Ia antigens, except for B lymphocytes, which are Ia(+) and still incapable of serving as accessory cells. Splenic adherent cells and resident peritoneal cells were both proficient at restoring anti-HSV CTL responses, although splenic adherent cells were clearly superior at limiting cell numbers. Neither population was capable of accessory cell activity, however, if it was pretreated with anti Ia antiserum plus complement or if anti-Ia serum was present during induction. Peritoneal cells lose virtually all of their membrane-associated Ia antigen after a brief period of in vitro culture (24 to 48 h). Cultured peritoneal cells, as well as P388D(1) cells (normally Ia(-)), can be induced to express Ia antigens within 48 h if they are cultured with concanavalin A-stimulated spleen cell supernatants. Ia(+) P388D(1) cells without the extraneous macrophage factor are not able to restore CTL responsiveness to HSV-1 in vitro, whereas Ia(+) cultured macrophages are fully competent accessory cells. When Ia(+) P388D(1) cells were supplemented with the macrophage-derived soluble factor interleukin 1, they displayed a modest, but significant, capacity to restore secondary anti-HSV CTL responses. In addition, glutaraldehyde-fixed, HSV-1-infected Ia(+) peritoneal cells, which could not restore the CTL response alone, were capable of providing accessory cell function if extraneous interleukin 1 was provided. In contrast, Ia(-) cultured peritoneal cells, Ia(-) P388D(1) cells, and various other Ia(-) cell lines were unable to participate in the generation of CTL even in the presence of interleukin 1. The adherent cell population would therefore appear to be making at least two essential contributions to the process of CTL development, namely, the secretion of interleukin 1 and the presentation of antigen in the context of membrane-associated Ia antigen. PMID- 6290392 TI - Immunocompetence of chickens during early and tumorigenic stages of Rous associated virus-1 infection. AB - A study was designed to determine the effects of congenital infection with the Rous-associated virus-1 (RAV-1) on the immune function chickens during the early and late tumorigenic stages of infection. In another experiment, the effects of niridazole on the immune competence and the tumor incidence in chickens congenitally infected with RAV-1 were studied. Lymphocyte stimulation by phytohemagglutinin, the phytohemagglutinin skin test, the response to immunization with sheep erythrocytes and Brucella abortus, and histological evaluation of lymphoid organs were used to determine the immune competence in normal and infected chickens. Results indicated that both B- and T-cell immune functions during the early and late stages of RAV-1 infection were comparable to those of normal uninfected chickens. Administration of niridazole to congenitally infected chickens at 5 weeks of age for 7 or 21 days had no effect on the T-cell mediated immunity; however, administration of the drug for 21 days eliminated lymphoma development. Unlike infection with other oncogenic viruses such as those causing Marek's disease and reticuloendotheliosis, infection with RAV-1 caused no detectable immunodepression during the early and late stages of infection. PMID- 6290393 TI - In vivo and in vitro models of demyelinating disease: endogenous factors influencing demyelinating disease caused by mouse hepatitis virus in rats and mice. AB - Intracerebral inoculation of JHM virus (JHMV), the neuropathic strain of mouse hepatitis virus, into Wistar Furth, Wistar Lewis, and Fischer 344 rats at various ages indicated that Wistar Furth rats are more susceptible to the virus than are the other strains. Fischer 344 and Wistar Lewis rats were more resistant to inoculation at 2 and 5 days of age and completely resistant by 10 days of age. In contrast, Wistar Furth rats which were very susceptible at both 2 and 5 days of age remained susceptible until 21 days of age. Intracerebral challenge of an F1 cross between Wistar Furth and Wistar Lewis rats at 10 days of age indicated that resistance to JHMV infection is dominant. Cyclophosphamide treatment 28 days after intracerebral inoculation exacerbated an inapparent infection, leading to paralysis in eight of nine and death in six of nine Wistar Furth test rats. In such immunosuppressed animals, grey- and white-matter lesions were noted throughout the central nervous system, in contrast to the purely demyelinating lesions noted previously. Since rats, unlike mice, were not susceptible to disease after intracerebral injection with the serorelated viscerotropic strain MHV-3, we wished to extend our understanding of the neurological disease process elicited by the two viruses in rodents. For this reason, various mouse strains, including some with recognized immunodeficiencies, were challenged by different routes of inoculation. Intraperitoneal infection of nude and beige mice with JHMV indicated that lack of natural killer cell functions does not markedly enhance the susceptibility to virus, whereas T-cell activity appears to be essential for resisting infection. JHMV and MHV-3 replication in peritoneal macrophages from highly resistant A/J mice was reduced in comparison with that noted in macrophages from susceptible C57BL6/J mice. An initial intraperitoneal inoculation of JHMV was able to protect C57BL6/J mice against fatal intracerebral challenge within 3 days, whereas A/J mice remained susceptible beyond day 3. The protective effect did not appear to result from increased levels of circulating interferon, preceded elevation in serum JHMV-neutralizing antibody titers, and persisted for at least several weeks after intraperitoneal inoculation. Based on the combined studies described here and on previous work by us and others, it appears that the factors influencing the outcome of coronavirus disease in rodents are age at inoculation, route of challenge, genetic constitution of the virus and host, and competence of the immune system, particularly cellular immunity involving T-cells. PMID- 6290394 TI - Genetic and molecular studies of plasmids coding for colonization factor antigen I and heat-stable enterotoxin in several escherichia coli serotypes. AB - Plasmids coding for colonization factor antigen I (CFA/I) and heat-stable enterotoxin (ST) were identified in 10 strains of human enterotoxigenic Escherichia coli. The strains, which belonged to serogroups O63, O114, O128, and O153, were isolated in Bangladesh, Latin America, Spain, and South Africa. Two strains produced heat-labile enterotoxin in addition to ST. CFA/I-ST plasmids were mobilized from two O128 strains into E. coli K-12 with the R factor R1-19K-. Like the prototype CFA/I-ST plasmid NTP113, mobilized previously from an E. coli O78 strain into K-12, these two plasmids were non-autotransferring. All 10 CFA/I ST plasmids were incompatible with NTP113 and had molecular weights ranging from 59 X 10(6) to 72 X 10(6). The molecular properties of seven of these plasmids were compared with those of six CFA/I-ST plasmids previously mobilized from O78 strains from Ethiopia, South Africa, and Bangladesh and with those of one plasmid coding for CFA/I, ST and heat-labile enterotoxin from a South African strain of serogroup O63. Digestion with the restriction endonuclease HindIII showed that several plasmids had very similar fragment patterns and two were identical. Generally, a larger proportion of HindIII fragments were of common size in digests of plasmids identified in strains from related geographical areas, regardless of serogroup. However, all except one plasmid shared five or six HindIII fragments of the same size, one of which had been shown previously to be involved in CFA/I production. There was at least 90% DNA homology between CFA/I ST plasmids with a molecular weight of about 58 X 10(6) from O78 strains from different sources. Most of the DNA sequences of these plasmids were present in a larger CFA/I-ST plasmid (72 X 10(6) from an O128 strain. The results of genetic and molecular studies suggest that CFA/I and ST production is determined by very similar plasmids in different serogroups of human enterotoxigenic E. coli from several sources. PMID- 6290395 TI - Influence of antibodies to mannophosphoinositides on phospholipid synthesis in Mycobacterium smegmatis ATCC 607. AB - Antiserum to mannophosphoinositides had a significant inhibitory effect on phospholipid synthesis in Mycobacterium smegmatis ATCC 607 at different phases of growth; however, the inhibition was more pronounced in the exponential phase. The rate of synthesis of total phospholipids and individual phospholipids, viz., cardiolipin, phosphatidylethanolamine, and mannophosphoinositides, decreased in the presence of antiserum as compared with normal serum. The maximum inhibitory effect of antiserum was exerted on the synthesis of mannophosphoinositides, particularly on tetraacylated dimannophosphoinositide. Antiserum also had a growth inhibitory effect as compared with normal serum. PMID- 6290396 TI - Seroepidemiology of heat-labile enterotoxigenic Escherichia coli and Norwalk virus infections in Panamanians, Canal Zone residents, Apache Indians, and United States Peace Corps volunteers. AB - Serum antibody titrations against the heat-labile enterotoxin (LT) of Escherichia coli were carried out on Panamanians, U.S. citizens resident in the Panama Canal Zone, Apache Indians living on the reservation in Whiteriver, Arizona, and Peace Corps volunteers before they traveled overseas. Antibody titers to Norwalk virus were also carried out on serum from Panamanian and Canal Zone residents. A high prevalence of low-titer LT antibodies was found in infants and adults from Panama, the Canal Zone, and Whiteriver. Panamanian children aged 1 to 5 years had the highest LT antibody titers. Peace Corps volunteers had a low prevalence and titer of LT antibodies. Prevalence and titer of antibodies to Norwalk virus were generally higher in Panamanians compared with Canal Zone residents of the same age. In the populations we studied, various modes of transmission and mechanisms of immunity likely explain the differences which we observed in antibody prevalence and titer to these two enteric pathogens. PMID- 6290398 TI - Inhibition of mitogen-induced human lymphocyte responsiveness by polymixin antibiotics. AB - Polymixin antibiotics, polymixin B and polymixin E (colistin) inhibited the mitogen-induced lymphoproliferative response of human lymphocytes. Inhibition of the lymphocyte response to PHA, PWM and Con A was evident at a low concentration of 1 U/ml of antibiotics. Lymphocytes in which the signals for proliferation had occurred were similarly prevented from proliferating. The effects were not due to cell death (toxicity). Since polymixin concentrations at which inhibition of lymphocyte proliferation was observed are employed in tissue culture medium and are also attained in plasma of patients, the results suggest that the use of the antibiotics in lymphocyte cultures limits lymphocyte responsiveness and that patients receiving polymixin antibiotics may experience a state of immunosuppression. PMID- 6290397 TI - Early interactions of herpes simplex virus with mouse peritoneal macrophages. AB - Adsorption of herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2) to resident peritoneal macrophages (PM) of 4-week-old Swiss albino (SA) and GR/AFib mice was studied. A significantly (P less than 0.05) higher HSV-2 adsorption rate was found with PM of SA mice than with PM of GR/AFib mice. Of added HSV-2 65% bound to the cells of SA mice over a 120-min period versus 15% to PM of GR/AFib mice. Only 15 to 20% of added HSV-1 bound to PM regardless of the mouse strain. These patterns of adsorption were found with all four HSV-1 and four HSV-2 strains tested. Pretreatment of PM with an HSV-2 mutant blocked the adsorption of added HSV-2. Thus, the receptors for HSV attachment seemed to be virus type selective. To avoid masking of adsorption by phagocytotic activity, the adsorption studies had to be performed at 4 degrees C. Transport of attached HSV-1 and HSV-2 to the nuclei of SA PM was studied with purified virus labeled with 32Pi and [3H]thymidine. In double-isotope experiments, only transport of HSV-2 was detected. The possible importance of differences in density or avidity of virus binding receptors on the plasma membrane of PM is discussed in relation to macrophage-dependent focal liver necrosis, which was only demonstrable after intraperitoneal inoculation of HSV-2, not HSV-1, only in SA, not GR/AFib, mice. PMID- 6290399 TI - Role of 3'-end of viral genome in tumor heteroinduction by the avian sarcoma virus. AB - Transformation defective virus was derived by restriction endonuclease cleavage from a clone of the avian sarcoma virus Schmidt-Ruppin strain, strongly oncogenic for rats. The transfection experiments of chicken cells by digested proviral DNA gave rise to transformation defective virus. The td virus was possible to recover in vivo in chickens. The tumors obtained after a long latent period contained the sarcoma virus which was able to transform chicken cells in vitro and to induce tumors in chickens. All viruses, parental, td- and recovered were of D subgroup specificity. The tumor induction experiments in rats have shown that the recovery of viral genome deletion in td mutant by cellular sequences was not enough to regain the oncogenicity for rats. The results stressed the importance of 3-end sequences of the virus genome, probably the sequences in C region for heteroinduction ability of the avian sarcoma virus. PMID- 6290400 TI - Leukemogenic activity of B-ecotropic C-type retroviruses isolated from tumors induced by radiation leukemia virus (RadLV-RS) in C57BL/6 mice. PMID- 6290401 TI - The human type-C retrovirus, HTLV, in Blacks from the Caribbean region, and relationship to adult T-cell leukemia/lymphoma. AB - Type-C RNA tumor viruses have been implicated in the etiology of naturally occurring leukemias and lymphomas of animals. Human T-cell leukemia/lymphoma virus (HTLV) is the first human virus of this class consistently identified in association with a specific type of human leukemia/lymphoma. The isolation of HTLV was made possible by the ability to grow mature T-cells in tissue culture usually with T-cell growth factor (TCGF). We now report a cluster of adult T-cell leukemia/lymphoma among Blacks from the Caribbean in which all eight cases are positive for HTLV virus and/or antibody. These patients have disease that appears indistinguishable from Japanese adult T-cell leukemia/lymphoma which, as we have also reported, is associated with HTLV in over 90% of cases. The finding of HTLV antibodies in some of the normal population in the Caribbean and Japan, and the clustering of a specific form of T-cell leukemia/lymphoma in these virus-endemic areas, suggest that HTLV infection may be associated with the occurrence of a distinctive clinico-pathologic entity. PMID- 6290402 TI - Cyclic AMP mediated modulation of complement protein production. AB - We have investigated the mechanisms by which cAMP analogues and phosphodiesterase inhibitors, reduced the production of C2 by monocytes in culture. Pulse label studies with 3H-labelled aminoacids showed that dibutyryl cAMP (dbcAMP) impaired the secretion of newly synthesised protein, both total (acid-precipitable) and individual complement proteins (precipitated antibody by antisera to C4, C2, C3, C5, B, P, C3b inactivator and beta 1H). The intracellular degradation of newly synthesised protein was increased in dbcAMP-treated cultures and protein synthesis was reduced. Studies aimed at defining the temporal relationships between these changes showed that protein secretion was impaired on the first day of culture, and increased degradation of newly synthesised protein was obvious by day 2. Protein synthesis was not significantly reduced until day 3 of culture. It is proposed that changes in intracellular cAMP levels may act as a second signal in the control of protein production by monocytes. PMID- 6290404 TI - Incidence and secular trends of congenital limb defects in Finland. AB - The total incidence rate of three selected congenital limb defects was 12.8 per 10 000 births in Finland from 1964 to 1977; 1235 cases were reported to the Registry of Congenital Malformations. The incidence rate of reduction limb deformities was 5.0 per 10 000 births (481 cases), of polydactyly 5.8 (559) and of syndactyly 3.1 (299), respectively. The total incidence rate and the rates for all the subgroups had a statistically significant increasing trend even after the incidence rates were compared to the reporting rate of all malformations (p less than 0.01). No significant variations in seasonal distributions were found. In 69% of the reduction deformities only one or both upper limbs were affected, 20% of the children had only lower limb defects, and 11% of the cases had defects in both upper and lower limbs. In 28% of the cases additional malformations were reported. For comparison, incidence rates of the selected limb defects from 13 other national surveillance systems are presented. PMID- 6290403 TI - Oral ketoconazole in the treatment of leishmaniasis. PMID- 6290405 TI - Prevalence of hepatitis markers in Roman children. AB - A random sample of sera from children admitted over a 23-month period to a large paediatric hospital in Rome for diagnoses believed unrelated to viral hepatitis, was tested for HBsAG, anti-HBs, anti-HBc and anti-HAV. No sex or area of residence differences were found. The prevalence for markers of prior hepatitis B infection in children aged 1-12 years was 6.5%, in agreement with other observations in Italy and European countries. Prevalence of anti-HAV among children age 1-12 admitted for diagnoses believed unrelated to viral hepatitis was 10%. Prevalence of hepatitis markers in Roman women of reproductive age is estimated as 23.5 for B and 68.2 for A. PMID- 6290407 TI - Ephedrine, a potential slimming drug, directly stimulates thermogenesis in brown adipocytes via beta-adrenoreceptors. AB - Ephedrine is a potential slimming drug that stimulates thermogenesis in man and laboratory animals. Considering that brown adipose tissue is an important site of catecholamine-induced thermogenesis in homeotherms, we tested the thermogenic efficiency of several ephedrine stereoisomers on adipocytes isolated from rat interscapular brown adipose tissue. Addition of (-)-ephedrine (0.1 mM) to brown adipocyte suspensions rapidly stimulated cellular respiration eight times above basal values. A stable Vmax of 335 nmol O2/min/10(6) cells was reached less than 5 min after the onset of respiratory stimulation. This value represents 85 percent of the maximal respiration observed with norepinephrine, the physiological effector of thermogenesis. The (-)isomer of ephedrine (1/2 Vmax = 20 microM) was more potent that other stereoisomers (+)-psi-ephedrine, (-)-psi ephedrine (racephedrine) in enhancing brown adipocyte respiration. Beta Adrenergic antagonists (alprenolol and propranolol) were much more effective than alpha-adrenergic antagonists (phentolamine and phenoxybenzamine) in inhibiting the respiratory effects of ephedrine. It is concluded that (-)-ephedrine mimics the calorigenic action of norepinephrine by directly stimulating brown adipocyte respiration via beta-adrenoreceptors. PMID- 6290406 TI - Some aspects of the human carcinogenicity of aflatoxin B1 and 3, 4 benz [a] pyrene. PMID- 6290408 TI - beta-Endorphin: synthesis and properties of human beta-endorphin-(1--28) and its analogs. AB - Three analogs of human beta-endorphin (beta h-EP) were synthesized by the solid phage method: beta h-EP-(1--28) (I). [Gln8]-beta h-EP-(1--28) (II), and [D-Ala2, Gln8]-beta h-EP-(1--28) (III). Radioreceptor binding assay with use of tritiated beta h-EP as primary ligand gave relative potencies as follows: beta h-EP, 100; I, 85; II, 380; III, 146. Relative potencies in an analgesic assay were: beta h EP; 100; I, 18; II, 36; III, 13. PMID- 6290409 TI - Dehydro-enkephalins. VI. Dehydroalanine3-enkephalin: a potent enkephalin analog for the delta opiate receptor. AB - The Gly3 residue in Gly2- and D-Ala2-enkephalins has been replaced by delta Ala3 and Ser3 in order to examine the effect on binding to the delta and mu opiate receptors. [D-Ala2, delta Ala3, Leu5]-enkephalin maintains most of its receptor binding affinities for both sites. It is suggested that the proper conjunctions of positions 2 and 3 of the enkephalin sequence are important to receptor preference and that position 3 may have a very specific interaction with the delta receptor. The in vivo analgesic and CNS activities of the peptides are also discussed. PMID- 6290410 TI - Visible-light-induced OH radicals in DNA-proflavine complexes: an e.p.r. and spin trapping study. AB - Using the spin-trapping technique, irradiation with visible light of complexes between DNA and proflavine was shown to generate OH radicals. The characteristic spectra were not obtained when proflavine or DNA were irradiated alone, nor when oxygen was absent. Using DMPO as a spin trap we found that the intensity of the DMPO-OH e.p.r. signal was enhanced when the molar ratio between bound proflavine and the DNA phosphorus increased up to a value of 0.15 demonstrating the efficiency of the intercalated dye molecules. A strong decrease of the e.p.r. signal was observed in the presence of various OH. scavengers like t-butanol, isopropanol or sodium benzoate. The OH. production was also decreased when the irradiation was made in the presence of SOD, ceruloplasmin or catalase and after addition of Chelex 100 resin. PMID- 6290411 TI - Oxygen-dependent damage involving OH radicals in irradiated bacteria. PMID- 6290412 TI - Spin-trapping studies of hydroxyl radical addition to cytosine and 2' deoxycytidine. PMID- 6290413 TI - Young woman with Cushing's syndrome. PMID- 6290414 TI - Radionuclide measurement of differential glomerular filtration rate in urinary tract obstruction. AB - In a previous study using dogs whose renal function was rendered asymmetric by unilateral infarction, the efficacy of technetium-99m (99mTc) DTPA and DMSA in measuring differential glomerular filtration rate (GFR) was demonstrated. The present study was undertaken to determine whether the same techniques were applicable to unilateral ureteral obstruction. Five normal dogs and nine dogs with partial unilateral ureteral obstruction had determination of glomerular filtration rate by standard techniques using constant infusions of iothalamate and creatinine after ureteral catheterization. These results were compared with total GFR as measured by single injection of 99mTc DTPA and analysis of the plasma disappearance curve. Calculated differential GFR was obtained by multiplying total GFR from double exponential analysis of this curve (DTPA2) by each of three measures of differential function. These included the percent differential uptake of 99mTc DTPA and 99mTC DMSA in the posterior projection as well as the geometric mean of 99mTc DMSA uptake. There were good correlations between differential GFR determined by iothalamate clearances at ureteral catheterization and all noninvasive methods involving radionuclides and DTPA2 ( r = 0.93-0.99). Single exponential analysis of the 99mTc DTPA plasma disappearance curve was less satisfactory than double exponential analysis. These results and those reported previously support the use of radionuclides in the determination of differential GFR in a variety of clinical situations. PMID- 6290415 TI - Influence of cell culture conditions on the inhibition of herpes simplex virus type 1 replication by acyclovir. AB - Inhibition of herpes simplex virus type 1 (HSV-1) replication, as measured by plaque reduction by acyclovir (ACV), was studied with different cell cultures and under varying conditions. Human lung fibroblasts (HL) required much lower concentrations of ACV for inhibition of virus replication than green monkey kidney (GMK) cells. In both cell types, ACV had to be added within 7 h after infection to cause a full antiviral effect. Pretreatment of cells with ACV before infection did not increase the antiviral activity. ACV caused a stronger inhibition in HL cells which had been confluent for 4 or 7 days as compared with cells just reaching confluence. Addition of ACV and its subsequent removal caused an irreversible plaque reduction in our experiments. ACV gave a pronounced inhibition of thymidine kinase (TK)-positive HSV-1 strains. A relatively small inhibitory effect was seen with a TK-negative HSV-1 strain in HL cells, and no measurable inhibition was seen in GMK cells. Inhibition of HSV-1 replication by ACV was concluded to depend on the type of virus, the cell type used, and the condition of the cells. PMID- 6290416 TI - Characterization of SV40-transformed human liver cells before and after passage through crisis. AB - SV40-transformed human liver cells could be passed through crisis after transfection with various DNAs (SV40, BKV, or salmon sperm). The pre- and post crisis cells were compared with respect to growth behavior and the state of the viral genome. Comparison of the viral inserts in the pre- and post-crisis cells showed minor changes in the integration pattern of the SV40 DNA after recovery from crisis. PMID- 6290417 TI - Heat-resistant infectivity of herpes simplex virus revealed by viral transfection. AB - Following apparently effective inactivation of herpes simplex virus types 1 and 2 at 56 or 70 degrees, significant infectivity was found upon viral transfection using the calcium-dimethylsulfoxide technique. Infectivity was quantitated in RSB 1 (rabbit skin fibroblast) and BHK-21 cells, and dose-response curves were found to depend on both the cell system and side effects of heat. Titers were dependent upon thermal treatment, as well as initial infectivity and cell origin of virus, and showed no or only moderate decrease after 6-hour exposures. There was no liberation of DNA or DNA-protein complexes, since infectivity was DNase- and trypsin-DNase-resistant. Upon purification, activity was found associated with particles that had retained their gross physical integrity. Obviously, protein(s) involved in initiation of infection was primarily altered. Heated virus was also insensitive to antiserum, which prevented replication of unheated, transfected virus. PMID- 6290418 TI - Isolation of a Golgi rich fraction from rat ventral prostate. AB - Unmodified procedures for isolation of fractions rich in Golgi elements from other tissues have not proved applicable to the rat ventral prostate because of the tendency of membranous material to aggregate. We have devised a new procedure whereby: 1) a Golgi rich fraction from rat ventral prostate was released by a gentle two-step homogenization and isolated by centrifugation through discontinuous sucrose density gradients; 2) the specific activity of UDP galactose: glycoprotein galactosyltransferase increased 69-fold in this fraction; 3) the isolated Golgi fraction was reasonably free from mitochondria, lysosomes, endoplasmic reticulum and plasma membranes as shown by the relatively low activities of marker enzymes; 4) the specific activities of acid phosphatase and 5'-nucleotidase in the Golgi rich fraction was 4 times greater than that in prostate homogenate. Both enzymes are secretory products and their presence in Golgi elements is probably associated with their packaging in secretory granules. PMID- 6290419 TI - Membrane fluidity and cholesterol in thymus and spleen cells from mice treated with immunomodulatory drugs. AB - We have used spin labeling, fluorescence polarization, and chemical analysis to characterize membrane properties of thymocytes from mice treated with immunomodulatory drugs. The number of thymocytes was reduced 90-95% by treatment of 6-9 week old mice with hydrocortisone acetate (HCA) or methylprednisolone (both 125 mg/kg) or with cyclophosphamide (250 mg/kg). Electron spin resonance (esr) examination of thymocytes labeled with 5-nitroxyl stearic acid indicated that the membranes of cells remaining after treatment with any of these drugs were more rigid than those from saline-treated controls. The total cholesterol/phospholipid (C/PL) molar ratio of the HCA-resistant thymocytes was twice that of the control mice. Treatment of mice with other immunomodulatory drugs, cyclophosphamide, cytosine arabinoside (Ara-C), 2-amino-5-bromo-6-phenyl-4 (3H)-pyrimidinone (ABPP) and 15(S)-methyl prostaglandin E1 (15(S)-methyl PGE1), also altered the C/PL ratio in thymocytes and, in some cases, in spleen cells. Fluorescence polarization measurements of thymocytes labeled with 1,6-diphenyl 1,3,5-hexatriene (DPH) did not reveal the differences between cells from HCA-and saline-treated mice that were detected by spin labeling and chemical analysis. Our results indicate that the greater rigidity detected by spin labeling of hydrocortisone-resistant thymocytes may be due, at least in part, to greater membrane cholesterol content. Of the methods employed, chemical analysis was the most sensitive in revealing drug-induced alterations in thymocyte populations. PMID- 6290420 TI - Malignant tumours of the bones of the hand (from the records of the Rizzoli Institute). AB - The authors present ten cases of malignant neoplasm of the bones of the hand, four primary tumours and six solitary metastatic tumours, from the case material of the Rizzoli Institute. They stress the importance of differential diagnosis of these rare neoplasms, in particular from benign neoplastic or hyperplastic lesions, which are common in the hand. PMID- 6290421 TI - Soft tissue sarcoma of the hand. AB - The authors report twelve cases of soft tissue sarcoma distal to the flexor crease of the wrist. These were isolated from a total of 414 cases of soft tissue sarcoma treated at the Rizzoli Institute. This case material comprises three epitheloid sarcomas, four fibrosarcomas, two rhabdomyosarcomas, one synovial sarcoma, one myxoid malignant fibrous histiocytoma, and one myxoid chondrosarcoma. The authors deal in detail with the biological behaviour, the criteria of differential diagnosis, and the indications for treatment in each histological type of sarcoma. PMID- 6290422 TI - Determination of transverse shielding for proton accelerators using the Moyer model. PMID- 6290423 TI - Dose calculations for the respiratory tract from inhaled natural radioactive nuclides as a function of age--II. Basal cell dose distributions and associated lung cancer risk. AB - Dose calculations for inhaled radon decay products presented in Part I (Ho79) have revealed that the doses to tracheobronchial and pulmonary compartments of the ICRP lung model are significantly dependent on age. From a consideration of the nonuniform dose distribution within the tracheobronchial region, doses are now calculated for the bronchial epithelium basal cells which are commonly regarded as the critical target for the induction of lung cancer. For the simulation of deposition and clearance mechanisms a refined mathematical model for postnatal growth of the human respiratory tract was developed on the basis of the Weibel model A. A reference atmosphere of 1 pCi/l for each nuclide with a mean respiratory minute volume, corresponding to a mean physical activity, was used to determine doses for the basal cells in different generations of the tracheobronchial tree as functions of age. The results obtained show again a strong dependence on age. In general a continuous decrease of dose with increasing age can be seen, with distinct differences between the various generations. If, however, the physical activity distribution and the ratio for the decay products as already defined in Part I are used, maximum dose values appear again in all generations at the age of about six years. Application of data on the relative risk of the induction of various malignancies versus age, taken from the BEIR report, results in even more pronounced dose maxima with a significantly higher radiation risk for children between birth and ten years of life of about one order of magnitude as compared to adults. PMID- 6290424 TI - Dose conversion factors for radon daughters in underground and open-cut mine atmospheres. AB - As uranium will be mined in Australia by open-cut methods, previous work on calculating dose conversion factors for radon daughters has been re-examined. The fractions of radon daughters deposited on lung airways and the factors for converting from equilibrium activity of radon daughters on airways to dose to basal cells are re-calculated. The variation around these estimates through variability of lung morphology and the depth of the basal cells is discussed. Average dose conversion factors calculated for atmospheres which may be typical of underground mines range from 12 mGy/WLM to 33 mGy/WLM. Use has been made of measurement results on unattached fraction at an open-cut mine in the Northern Territory, Australia, to derive dose conversion factors, ranging from 50 mGy/WLM to 135 mGy/WLM which are applicable to this environment. PMID- 6290425 TI - The activity of radon daughters in high-rise buildings. PMID- 6290426 TI - The tissue distribution and excretion of actinides absorbed from the gastrointestinal tract of rodents. PMID- 6290427 TI - Nosocomial urinary tract infections. PMID- 6290428 TI - Southern blot hybridization in cetaceans, using killer whale restriction fragment as a probe. PMID- 6290429 TI - Human papilloma virus infection (condyloma) of the uterine cervix in Japanese women. PMID- 6290430 TI - Ultrastructural cytochemistry of phosphatases in the ductal component of pleomorphic adenoma of human parotid and submandibular salivary glands. AB - Some ductal cells of pleomorphic adenomas showed evidence of secretory activity, with apical secretory granules, thiamine pyrophosphatase activity in the Golgi apparatus, and acid phosphatase activity in GERL-like structures and in immature secretory granules. Alkaline phosphatase activity was demonstrated rarely at the luminal plasma membrane and in intracellular vesicles, suggesting resorptive activity. ATPase reaction product was associated with contiguous surfaces of tumour cells, particularly of those cells adjacent to the basement membrane, these latter cells apparently differentiating in a different manner to the luminal cells. A comparison of luminal ductal cells of the tumours with normal salivary glands revealed most similarity with intercalary ductal cells. PMID- 6290431 TI - Histochemical studies of fibroblasts from patients with Menkes kinky hair disease and Wilson's disease. AB - Cultured fibroblasts from Menkes kinky hair disease patients showed markedly reduced succinate dehydrogenase and amine oxidase activities. Cytochrome oxidase activity, however, was greatly reduced in some cells and almost normal in others. Cultured fibroblasts from patients with Wilson's disease showed moderately reduced succinate dehydrogenase and cytochrome oxidase activities. Amine oxidase activity was only slightly reduced when compared to that of normal. These results indicated that the histochemical phenotype observed in fibroblasts from patients with Menkes kinky hair disease and Wilson's disease were distinctly different from each other and from normal fibroblasts. PMID- 6290432 TI - Heterogeneity in polymorphonuclear leukocyte neutrophil granules. PMID- 6290433 TI - Dietary survey in 40-year-old Edinburgh men. AB - One hundred and seven healthy 40-yr-old Edinburgh men were asked to participate in a dietary survey involving a 7-d weighed record. Ninety seven successfully completed the record. The results showed that the total energy was made up of 13 per cent protein, 38 per cent fat, 40 per cent carbohydrate and 9 per cent alcohol. A small proportion of the men had exceptionally high alcohol intakes. The mean intake of fat was high, 121 g, it is nevertheless lower than amounts consumed in other areas of Britain. Of the energy from fat, 16 per cent came from saturated-fatty acid, 15 per cent from monounsaturated-fatty acid and 4 per cent from polyunsaturated-fatty acid. Linoleic acid contributed 3 per cent of the mean energy intake. In some mens' diet this essential fatty acid provided under 2 per cent of their total energy consumption. PMID- 6290435 TI - Hemodynamic response to converting enzyme inhibition at rest and exercise in humans. AB - The response of the systemic circulation to acute inhibition of the converting enzyme with 25 mg of oral Captopril (Squibb) was studied in six normal sodium replete male volunteers at rest and during exercise, together with its effects on exercise capacity for graded uninterrupted exercise. In recumbent subjects at rest Captopril did not affect arterial pressure or heart rate, and plasma renin activity rose 2.5-fold (P less than 0.05). In subjects in the sitting position, at rest and during exercise until exhaustion, Captopril reduced mean brachial intra-arterial pressure by an average of 7 Torr in comparison to placebo (P less than 0.001). Captopril's hypotensive effect was caused by a reduction of systemic vascular resistance (P less than 0.01), without changes of cardiac output (measured by CO2 rebreathing), heart rate, or stroke volume. Plasma renin activity was significantly higher during Captopril (P less than 0.001). Peak oxygen uptake and exercise duration were the same after administration of Captopril or placebo. The data demonstrate that the renin-angiotensin system is not involved in the homeostasis of blood pressure in supine sodium-replete humans, but has a modest role in blood pressure regulation when posture is changed from supine to upright. The orthostatic effect of Captopril is maintained during upright exercise. Furthermore the reduction of systemic vascular resistance by Captopril does not affect peak oxygen uptake. PMID- 6290434 TI - The localization of converting enzyme in kidney vessels of the rat. AB - An antibody against pure rabbit lung converting enzyme (CE) showing cross reaction with CE from other species was used for immunocytochemical studies in the kidney of rats. Using the indirect labelling PAP-technique, specific immunostaining was found in the endothelial layer of all arteries and arterioles of kidney cortex and in some descending vasa recta. CE-positive reactions were also seen in most glomeruli, the reaction product being confined to only a few capillary loops in connection with the glomerular stalk. A few immunostained capillaries in the cortical labyrinth were suspected to belong to the first ramifications of the efferent arteriole. The bulk of all other of the glomerular and peritubula capillaries as well as all veins of the kidney showed no obvious immunostaining. The functional significance of this specific localization pattern of CE in the endothelium of kidney vessels is discussed with respect to the actions of the systemic and the local, intrarenal renin-angiotensin-system on kidney functions. PMID- 6290436 TI - Hyperoxia inhibits stimulated superoxide release by rat alveolar macrophages. AB - Factors responsible for the loss of respiratory burst capacity (stimulated extracellular O2-. release) of alveolar macrophages (AM) exposed to prolonged hyperoxia were assessed. Specific pathogen-free rats were exposed to 1 ATA O2 for 24-72 h, and lungs of survivors lavaged. Release of O2-. by cells after addition of concanavalin A, which stimulated AM but not polymorphonuclear leukocytes (PMN), or digitonin, which stimulated both cell types, was measured using cytochrome c reduction +/- superoxide dismutase. O2-. release by AM declined 47.2% (P less than 0.05) after 24 h of hyperoxia and 100% after 60 h. Percent PMN in the lavage was less than 3% at 0-36 h but increased to 16% at 48 h and to 44% at 72 h. Although addition of PMN to AM in vitro caused inhibition of AM O2-. release, the percent PMN required for inhibition was not reached in vivo until after a significant decline in AM O2-.-releasing capacity had already occurred. Cell-free lavage fluid from either control or hyperoxic rats did not affect AM O2 . release. AM in culture for 24 h in hyperoxia lost 76.7% (P less than 0.005) of O2-.-releasing capacity vs. cells incubated in 20% O2, although dye exclusion was unaffected. The results indicate that the major cause of loss of AM O2-. release by hyperoxia is a direct effect of O2 on the cells. PMID- 6290438 TI - Raccoons are not susceptible to canine parvovirus. PMID- 6290437 TI - Colostral and milk antibody titers in cows vaccinated with a modified live rotavirus-coronavirus vaccine. AB - Colostral and milk whey rotavirus (RV) and coronavirus (CV) antibody titers stimulated in 15 beef heifers by vaccination with a modified live-RV-CV vaccine were compared with titers in 15 nonvaccinated heifers. Geometric mean antibody titers to RV in colostral and 3-day milk whey from vaccinated heifers were 2,807 and 92, respectively, and in control heifers were 1,613 and 71, respectively. Geometric mean antibody titers to CV in colostral and 3-day milk whey of vaccinated heifers were 877 and 13, respectively, compared with titers were 877 and 13, respectively, compared with titers in nonvaccinated heifers of 731 and 7, respectively. Differences in antibody titers between vaccinated and nonvaccinated heifers were not significantly different. PMID- 6290439 TI - Abomasal adenocarcinoma in a cow. PMID- 6290440 TI - The ACTH cells in the pituitary gland of the nine-spined stickleback, Pungitius pungitius L. AB - The ACTH cells form a layer 1-8 cells thick, dorsal to the prolactin cells in the rostral pars distalis. They react only mildly with a variety of stains including PAS-lead haematoxylin. Their nuclear diameters vary seasonally in a manner that closely parallels that of the prolactin cells. The relative volumes of the ACTH and prolactin cell zones are remarkably constant in animals of different sizes. It is suggested that the two hormones may act synergistically at various body sites and that this accounts for the related morphological features of the ACTH and prolactin cells. There are no changes in the surface density of the ACTH zone with increasing animal size. Consequently, the ACTH/neurohypophysial border is highly convoluted in large animals. The proximity of the neurohypophysis also influences cell ultrastructure, as small processes, packed with secretory granules are more numerous near the basal lamina separating the adeno- and neurohypophyses. A morphometric analysis of ACTH cells provides base-line ultrastructural data for experimental studies and for comparisons with other teleosts. The cells have small, secretory granules, 100-300 nm in diameter, and of variable electron density. There is little rough endoplasmic reticulum and a small Golgi apparatus. There is no evidence of granule release by exocytosis and various explanations for this are suggested. PMID- 6290441 TI - Crude fiber determination using ceramic fiber to replace asbestos. AB - Crude fiber was determined in a wide range of feed products by a method which specifies ceramic fiber as a filter medium instead of the more hazardous and difficult to obtain asbestos. Results correlated well with those obtained by using AOAC official final action method 7.061-7.065 (correlation coefficient, 0.9994). For 8 samples, the coefficients of variation ranged from 0.74 to 4.80%. Compared with the AOAC method the proposed method showed a slight negative bias of 0.1%. Compared with asbestos, ceramic fiber was easier to prepare for use, filtering was faster, and samples bumped less. PMID- 6290442 TI - Primary liver cell carcinoma. PMID- 6290443 TI - The radiological investigation of primary hepatic tumours in infancy and childhood. A report of 8 cases. PMID- 6290445 TI - Effect of 4,5',8-trimethylpsoralen interstrand cross-links present in recipient Bacillus subtilis on the integration of transforming DNA. AB - When recipient Bacillus subtilis carrying chromosomal trimethylpsoralen cross links were transformed, the donor marker activity decreased with the extent of cross-linking. Additional donor marker activity was lost upon incubation of the reextracted DNA with nuclease S1, particularly at higher levels of cross-linking. Physical analysis of the reextracted DNA showed that the donor DNA was progressively excluded from heteroduplex formation as the frequency of cross links in the recipient DNA increased. In the donor-recipient complexes still being formed, increasing amounts of donor DNA became susceptible to nuclease S1 digestion under these conditions. These results suggest that resident interstrand cross-links interfere both with initiation of recombination and with the completion of heteroduplex formation. PMID- 6290444 TI - Escherichia coli phenylalanyl-tRNA synthetase operon: characterization of mutations isolated on multicopy plasmids. AB - Plasmid pB1 carries the genes for threonyl-tRNA synthetase, phenylalanyl-tRNA synthetase, and translation initiation factor IF3. Strains carrying this plasmid overproduce phenylalanyl-tRNA synthetase about 100-fold. Spontaneous mutant plasmids were obtained which no longer caused the overproduction of the enzyme. Three classes of mutations were found. (i) Deletion mutations were found, some of which had the interesting property of fusing different genes together, e.g., putting phenylalanyl-tRNA synthetase under the control of the threonyl-tRNA synthetase promoter. (ii) Insertion mutations were found; one insertion in particular was studied. This insertion is located in front of the structural gene for phenylalanyl-tRNA synthetase and is shown to interrupt a cis-acting regulatory region. (iii) Mutations that showed no major change in DNA structure were found. One of these mutations is apparently purely structural, as it produces a small subunit of phenylalanyl-tRNA synthetase with a reduced molecular weight. This protein is less stable than the wild-type enzyme. These mutations represent useful tools to investigate how the phenylalanyl-tRNA synthetase operon is regulated. PMID- 6290446 TI - Enzymes of agmatine degradation and the control of their synthesis in Streptococcus faecalis. AB - Streptococcus faecalis ATCC 11700 uses agmatine as its sole energy source for growth. Agmatine deiminase and putrescine carbamoyltransferase are coinduced by growth on agmatine. Glucose and arginine were found to exert catabolite repression on the agmatine deiminase pathway. Four mutants unable to utilize agmatine as an energy source, isolated from the wild-type strain, exhibited three distinct phenotypes. Two of these strains showed essentially no agmatine deiminase, one mutant showed negligible activity of putrescine carbamoyltransferase, and one mutant was defective in both activities. Two carbamate kinases are present in S. faecalis, one belonging to the arginine deiminase pathway, the other being induced by growth on agmatine. These two enzymes have the same molecular weight, 82,000, and seem quite different in size from the kinases isolated from other streptococci. PMID- 6290447 TI - Cloning of and complementation tests with alkaline phosphatase regulatory genes (phoS and phoT) of Escherichia coli. AB - The regulatory genes of alkaline phosphatase, phoS and phoT, of Escherichia coli were cloned on pBR322, initially as an 11.8-kilobase EcoRI fragment. A restriction map of the hybrid plasmid was established. Deletion plasmids of various sizes were constructed in vitro, and the presence of phoS and phoT genes on the cloned DNA fragments was tested by introducing the plasmids into phoS64 and phoT9 strains for complementation tests. One set complemented only phoS64 but not phoT9; the other set complemented only phoT9 but not phoS64. We conclude that phoS64 and phoT9 mutations belong to different complementation groups and probably to different cistrons. The hybrid plasmid with the 11.8-kilobase chromosomal fragment also complemented the phoT35 mutation. A smaller derivative of the hybrid plasmid was constructed in vitro which complemented phoT35 but did not complement phoS64, phoT9, or pst-2. Our results agree with the suggestion that phoT35 lies in a different complementation group from phoS, phoT, or pst-2 (Zuckier and Torriani, J. Bacteriol. 145:1249--1256, 1981). Therefore, we propose to designate phoT35 as phoU. The effect of amplification of phoS or phoT on alkaline phosphatase production was examined. It was found that multiple copies of the phoS gene borne on pBR322 repressed enzyme production even in low phosphate medium, whether it was introduced into wild-type strains (partially repressed) or phoR (phoR68 or phoR17) strains (fully repressed), whereas the introduction of multicopy plasmids bearing the phoT gene did not affect the inducibility of the enzyme. PMID- 6290448 TI - Replication and incompatibility properties of plasmid pE194 in Bacillus subtilis. AB - pE194, a 3.5-kilobase multicopy plasmid, confers resistance to the macrolide lincosamide-streptogramin B antibiotics in Bacillus subtilis. By molecular cloning and deletion analysis we have identified a replication segment on the physical map of this plasmid, which consists of about 900 to 1,000 base pairs. This segment contains the replication origin. It also specifies a trans-acting function (rep) required for the stable replication of pE194 and a negatively acting copy control function which is the product of the cop gene. The target sites for the rep and cop gene products are also within this region. Two incompatibility determinants have been mapped on the pE194 genome and their properties are described. One (incA) resides within the replication region and may be identical to cop. incB, not located in the replication region, expresses incompatibility toward a copy control mutant (cop-6) but not toward the wild-type replicon. PMID- 6290450 TI - Molecular mapping of glyW, a duplicate gene for tRNA3Gly of Escherichia coli. AB - By the use of [5'-32P]tRNA3Gly from Escherichia coli as a hybridization probe, glyW was located on cloned fragments of the uvrC pgsA region of the bacterial chromosome. After determination of the sites of action of several restriction enzymes, glyW was found to be within approximately 300 base pairs of pgsA. The order of genes in this region is uvrC, pgsA, glyW, flaI. Comparison of the order of determined restriction sites with the sites predicted from the nucleotide sequence of tRNA3Gly indicates that the direction of transcription of glyW is counterclockwise on the circular E. coli map. PMID- 6290449 TI - An amber mutation in the gene encoding the beta' subunit of Escherichia coli RNA polymerase. AB - An Escherichia coli strain carrying an amber mutation (UAG) in rpoC, the gene encoding the beta prime subunit of RNA polymerase, was isolated after mutagenesis with nitrosoguanidine. The mutation was moved into an unmutagenized strain carrying the supD43,74 allele, which encodes a temperature-sensitive su1 amber suppressor, and sue alleles, which enhance the efficiency of the suppressor. In this background, beta prime is not synthesized at high temperature. Suppression of the mutation by the non-temperature-sensitive amber suppressor su1+ yields a protein which is functional at all temperatures examined (30, 37, and 42 degrees C). PMID- 6290451 TI - Chromosomal location of a Bacillus subtilis DNA fragment uniquely transcribed by sigma-28-containing RNA polymerase. AB - A fragment of the Bacillus subtilis chromosome containing a gene whose transcription is dependent on sigma-28-containing RNA polymerase has been genetically mapped by means of an integrable plasmid. This gene resides on the chromosome adjacent to the stage 0 sporulation locus spo0E between metC and citL. The gene was insertionally inactivated by cloning an internal EcoRI-HindIII fragment in the integrable plasmid pJH101 and by inserting the plasmid into the chromosome by transformation. Transformants bearing an inactivated gene were indifferent to this inactivation for both growth and sporulation. PMID- 6290452 TI - Biosynthesis of oxygen-detoxifying enzymes in Bdellovibrio stolpii. AB - Axenically grown Bdellovibrio stolpii (i.e., grown independently of the host) was examined for superoxide dismutase, catalase, and peroxidase activities. Kinetics of enzyme synthesis were determined for aerobically grown cultures and for cultures exposed to 100% oxygen. Enzymatic activities varied with the age of the culture. Normally grown cultures exhibited maximum activity during the first 10 h of growth and again as the stationary phase was approached, beginning at about 48 h. Polyacrylamide gel electropherograms of cell-free extracts revealed that B. stolpii contained one major band (1) and two minor bands (II, III) of superoxide dismutase activity. Each of these enzymes was inactivated by H2O2, indicating that they were iron-containing enzymes. Manganese-containing superoxide dismutase was not detected in B. stolpii. Increased oxygenation did not appreciably stimulate enzyme synthesis, for only superoxide dismutase was induced, reaching maximum activity at 10 h and then rapidly falling to normal levels. Superoxide dismutase appears to be the main enzymatic defense against oxygen toxicity in B. stolpii. Induction of superoxide dismutase with 100% oxygen was manifested as an increase in the intensities of the two minor bands of activity, suggesting that isozyme I is constitutive, whereas isozymes II and III are inducible. The induction of isozymes II and III by 100% oxygen was prevented by an inhibitor of protein biosynthesis, chloramphenicol. PMID- 6290453 TI - Integration and partial excision of a cryptic plasmid in Pseudomonas syringae pv. phaseolicola. AB - A virulent strain of Pseudomonas syringae pv. phaseolicola, a pathogen of the common bean Phaseolus vulgaris (L.), was shown to harbor a 98-megadalton cryptic plasmid, pMC7105. After exposure of this strain to the plasmid-curing agent mitomycin C, a colony was isolated which had no detectable extrachromosomal DNA. Hybridization of labeled pMC7105 probe to nitrocellulose filters containing Southern-blotted BamHI cleavage products of cellular DNA revealed that pMC7105 was integrated into the chromosome rather than cured from this strain. Imprecise excision of pMC7105 resulted in the formation of three smaller plasmids of 34, 50, and 58 megadaltons. BamHI and EcoRI fingerprint analyses revealed that these plasmids were excised from a common region of pMC7105. The BamHI fragments of pMC7105 which were not present in the excision plasmids remained integrated and could be detected by hybridization of pMC7105 probe to Southern-blotted cellular DNA from these strains. Certain chromosomal fragments also had homology with the pMC7105 probe. The excision plasmids were stably maintained and neither integration nor excision altered the pathogenicity of these strains. PMID- 6290454 TI - Molecular cloning of the spo0B sporulation locus in bacteriophage lambda. AB - The stage 0 sporulation locus spo0B has been mapped by transformation between the pheA and spoIVF loci. Analysis of the behavior of alleles of the spo0B locus in trpE26 merodiploid strains indicates that all of the known alleles of this locus comprise a single complementation group. The spoIVF88 mutation was found to reside in a separate complementation group. The chromosomal region surrounding and including the spo0B locus was cloned in the lambda vector Charon 4A. Extensive restriction endonuclease analyses of the inserts in these phage revealed that an EcoRI fragment of DNA of 2.3 kilobases had transforming activity for spo0B mutations. Examination of the physical and genetic maps of the locus suggested that the entire spo0B locus is contained within this fragment. Subcloning of restriction endonuclease fragments of the lambda inserts and transformation analyses allowed assignment of surrounding genetic loci to specific DNA fragments. PMID- 6290455 TI - The incompatibility product of IncFII R plasmid NR1 controls gene expression in the plasmid replication region. AB - The incompatibility properties of IncFII R plasmid NR1 were compared with those of two of its copy number mutants, pRR12 and pRR21. pRR12 produced an altered incompatibility product and also had an altered incompatibility target site. The target site appeared to be located within the incompatibility gene, which is located more than 1,200 base pairs from the plasmid origin of replication. The incompatibility properties of pRR21 were indistinguishable from those of NR1. Lambda phages have been constructed which contain the incompatibility region of NR1 or of one of its copy mutants fused to the lacZ gene. In lysogens constructed with these phages, beta-galactosidase was produced under the control of a promoter located within the plasmid incompatibility region. Lysogens containing prophages with the incompatibility regions from pRR12 and pRR21 produced higher levels of beta-galactosidase than did lysogens containing prophages with the incompatibility region from the wild-type NR1. The introduction into these inc lac lysogens of pBR322 plasmids carrying the incompatibility regions of the wild type or mutant plasmids resulted in decreased levels of beta-galactosidase production. For a given lysogen, the decrease was greater when the pBR322 derivative expressed a stronger incompatibility toward the plasmid from which the fragment in the prophage was derived. This suggested that the incompatibility product acts on its target to repress gene expression in the plasmid replication region. PMID- 6290456 TI - Ornithine-containing lipid of Bordetella pertussis that carries hemagglutinating activity. AB - The proposed structure of the ornithine-containing lipid of Bordetella pertussis is 3-hydroxyhexadecanoic acid amide-linked to ornithine and esterified to the second hexadecanoic acid. The aminolipid strongly agglutinates type A and B human erythrocytes. PMID- 6290457 TI - Identification of chromosomally integrated TOL DNA in cured derivatives of Pseudomonas putida PAW1. AB - Some plasmid-free Tol- strains derived from Pseudomonas putida PAW1 (which carries the TOL plasmid pWW0) have a segment of TOL DNA located chromosomally. Of three independently isolated strains, PAW86 had an integrated TOL segment of 16 kilobases and PAW85 had two copies of this segment in different chromosomal locations, whereas the chromosomal DNA of PAW82 showed no homology with the TOL plasmid. In cultures of the parental strain, it appears that a 56-kilobase TOL DNA segment is located chromosomally in some cells. PMID- 6290459 TI - Physical map of a plasmid from Caedibacter taeniospiralis 51. AB - Caedibacter taeniospiralis 51 carries at least two plasmids, pKAP51 and pKAP52. The smaller plasmid, pKAP51, contains 43 kilobase pairs. The larger plasmid, pKAP52, contains more than 110 kilobase pairs. Relative positions of recognition sequences for seven different restriction endonucleases were determined, and a physical map of pKAP51, consisting of a total of 28 restriction sites, was constructed. PMID- 6290458 TI - Expression of the transposable lac operon Tn951 in Rhodopseudomonas sphaeroides. AB - The transposon Tn951 (lac) was introduced into the photosynthetic bacterium Rhodopseudomonas sphaeroides 2.4.1, which is normally Lac-, via the P-group plasmid RP1. beta-Galactosidase was produced constitutively in both chemotrophically and phototrophically grown cells, and the levels were found to be the same but low. Mutants were isolated, however, that were able to grow on lactose minimal medium and which expressed different levels of beta-galactosidase when grown chemotrophically or phototrophically. The beta-galactosidase levels found in all R. sphaeroides strains were much less than those found in Escherichia coli. PMID- 6290460 TI - Hybridization studies reveal homologies between pBF4 and pBFTM10, Two clindamycin erythromycin resistance transfer plasmids of Bacteroides fragilis. AB - Two clindamycin-erythromycin resistance transfer factors of Bacteroides fragilis, pBF4 and pBFTM10, were analyzed for regions of DNA homology. Although the plasmids were derived from different clinical isolates of B. fragilis and have different sizes, they showed homology in the clindamycin-erythromycin resistance region; no homology could be detected outside of this region. PMID- 6290461 TI - Cloning of the trp-1 gene from neurospora crassa by complementation of a trpC mutation in Escherichia coli. AB - Studies with a hybrid plasmid containing 4.0 kilobase pairs of Neurospora crassa DNA cloned into plasmid pBR322 indicated that the plasmid restored to prototrophy a trpC mutant of Escherichia coli which lacked phosphoribosyl anthranilate isomerase but not a trpC mutant which lacked indole glycerol phosphate synthetase, that the relevant transcription was initiated at a promoter within the N. crassa DNA, and that the phosphoribosyl anthranilate isomerase could be specified by a subcloned segment of the original DNA. PMID- 6290462 TI - Calcium-induced weakening of skeletal muscle Z-disks. AB - Structural changes in the Z disk were sensitively detected by measuring fragmentation indexes of myofibrils. The Ca2+-induced weakening of Z disks and the Z-disk removal by muscle calpain could be clearly distinguished by using muscle calpastatin, an endogenous inhibitor of muscle calpain. The Ca2+-induced weakening of Z disks occurred without concomitant release of alpha-actinin and had maxima at 10(-4) M Ca2+ and 45 degrees C and a minimum at pH 6.5, while the Z disk removal by calpain had similar optima to the caseinolytic activity of calpain, at 10(-3) M Ca2+, 20 degrees C and pH 7.0. The Ca2+-induced weakening of Z disks is therefore not due to the proteolytic action of calpain. In postmortem muscle, moreover, the Ca2+-induced weakening of Z disks was inferred to be predominate over calpain proteolysis, and therefore to be the major factor in the characteristic weakening of Z disks. PMID- 6290463 TI - Characterization of the Ca2+-gated cation channel in sarcoplasmic reticulum vesicles. AB - Two types of sarcoplasmic reticulum (SR) vesicles, i.e., normal (NSR) and heavy (HSR), were prepared by using a centrifugation technique, and further characterization of the cation transport in the two types of SR vesicles was performed by measuring the choline influx. The choline influx in SR vesicles was measured by following the change in light scattering intensity using a stopped flow apparatus. Analysis of the choline influx gave the following results. (1) There exists a Ca2+-gated cation channel in SR vesicles other than the K+ selective, voltage-dependent cation channel. The Ca2+-gated cation channel was activated by both extravesicular Ca2+ and caffeine, and inhibited by procaine and N,N'-dicyclohexylcarbodiimide. (2) About 70% of the HSR vesicles have Ca2+-gated cation channels, whereas only 20% of NSR vesicles have these channels. On the other hand, 60% of HSR vesicles also have the voltage-gated cation channels and 80% of NSR vesicles have these channels. On the basis of various properties of the Ca2+-gated cation channel, it is suggested that this channel must be the one which mediates the Ca2+-induced Ca2+ release. PMID- 6290464 TI - Kinetics of the actions of caffeine and procaine on the Ca2+-gated cation channel in sarcoplasmic reticulum vesicles. AB - The effects of caffeine and procaine on the Ca2+-gated cation channel in sarcoplasmic reticulum (SR) vesicles were studied by measuring choline influx. The choline influx in SR vesicles was measured by following the change in light scattering intensity using a stopped flow apparatus. From the kinetic analysis of the rate of choline influx, the following results were obtained. (1) The rate of choline influx was enhanced when Ca2+ bound to the Ca2+-receptor site of the Ca2+ gated cation channel. (2) Caffeine enhanced the choline influx by increasing only the affinity of Ca2+ for the receptor site of the channel and thus regulated the equilibrium between open and closed states of the channel. The affinity increased about 14-fold upon caffeine binding. The dissociation constant of caffeine was 10 mM. (3) In contrast, procaine itself blocked the choline influx mediated by the Ca2+-gated cation channel. The blockade followed a single-site titration curve with a Ca2+-dependent dissociation constant of 0.44 mM at 2 x 10(-6) M Ca2+. The Ca2+-dependence was explained by assuming that procaine would bind to the inhibitory site only when the channel was open. (4) Procaine also inhibited the choline influx enhanced by caffeine. The blockade could be explained on the basis of the above kinetic model. PMID- 6290465 TI - Mechanism and function of the Ca2+-gated cation channel in sarcoplasmic reticulum vesicles. PMID- 6290466 TI - Antibody to the nonapeptide Glu-Glu-Glu-Glu-Tyr-Met-Pro-Met-Glu is specific for polyoma middle T antigen and inhibits in vitro kinase activity. AB - Middle T antigen of polyoma virus has an associated tyrosine kinase activity which phosphorylates tyrosine residue 315 on middle T in immunoprecipitates. A peptide representing the sequence of middle T from residue 311 to 319 has been synthesized. This peptide acts as a weak inhibitor of the kinase reaction. An antiserum has been raised against this peptide after conjugation to bovine serum albumin. The antibody is middle T-specific. Middle T antigen precipitated by this serum is largely inactive in the kinase reaction. Dissociation of the immune complex with peptide releases middle T in a kinase-active form. PMID- 6290467 TI - Evidence for two structurally different forms of skeletal muscle Ca2+-activated protease. AB - Recent studies on calcium-activated protease (CAF) have indicated that there are two forms of this enzyme, one requiring millimolar levels of Ca2+ and one requiring micromolar levels of Ca2+ for maximal activation. We have attempted to elucidate the biochemical nature of the difference between the two forms by the use of one dimensional peptide maps and immunoautoradiography, and have found that the 80,000-dalton subunits from the two forms differ substantially while the 30,000-dalton subunit appear to be identical. PMID- 6290468 TI - Regulation of uridine kinase quaternary structure. Dissociation by the inhibitor CTP. AB - Uridine kinase from mouse Ehrlich ascites cells can exist in a variety of different aggregation states, from monomer up to aggregates that may contain 32 or more subunits. With very crude enzyme preparations, uridine kinase activity is always associated with several different coexisting molecular weight species. Changes in the aggregation state are produced in the presence of normal effectors (orthophosphate, ATP and CTP) at physiological concentrations. With uridine kinase that has been purified 9,000-fold, enzyme activity is associated with only a single molecular weight species, but is still responsive to the same physiological effectors. In the presence of orthophosphate, uridine kinase has a molecular weight of 380,000 (appropriate for a dodecamer). In the presence of CTP, the enzyme dissociates with concomitant loss of activity. The dissociated enzyme can be reassociated to the native size. These results imply that alteration of the enzyme's quaternary structure by normal effectors constitutes a mechanism for regulating uridine kinase activity in vivo. PMID- 6290469 TI - Mossbauer study of a bacterial cytochrome oxidase: cytochrome c1aa3 from Thermus thermophilus. AB - Cytochrome c1aa3 from Thermus thermophilus has optical and EPR properties similar to bovine cytochrome c oxidase. We have studied 87Fe-enriched samples with Mossbauer spectroscopy in the fully oxidized and fully reduced states and in the oxidized state complexed with cyanide. The cytochromes a and c1 yielded spectra quite similar to those reported for the cytochromes c and b5; in the oxidized state the spectra reflect noninteracting, low spin ferric hemes, whereas the a- and c1-sites of the reduced enzyme are typical of low spin ferrous hemochromes. The spectra of the reduced enzyme show that reduced cytochrome a3 is high spin ferrous, with Mossbauer parameters quite similar to those of deoxymyoglobin. Upon addition of cyanide to the oxidized enzyme, the a3-site exhibits in the absence of an applied magnetic field and at temperatures down to 1.3 K a quadrupole doublet with parameters typical of low spin ferric heme-CN complexes. The low temperature spectra taken in applied magnetic fields show that the electronic ground state of the a3-CN complex has integer electronic spin, suggesting ferromagnetic coupling of the low spin ferric heme (S = 1/2) to Cu2+ (S = 1/2) to yield as S = 1 ground state. We have examined the oxidized enzyme from two different preparations. Both had good activity and identical optical and EPR spectra. The Mossbauer spectra, however, revealed that the a3-site had a substantially different electronic structure in the two preparations. Neither configuration had properties in accord with the widely accepted spin-coupling model proposed for the bovine enzyme. PMID- 6290471 TI - Gene for Escherichia coli glycyl-tRNA synthetase has tandem subunit coding regions in the same reading frame. AB - Glycyl-tRNA synthetase is one of two Escherichia coli aminoacyl tRNA synthetases which has two different subunits. A 5.1-kilobase pair HindIII chromosomal DNA fragment was isolated, cloned into pBR322 (to give plasmid pTK201), and shown to direct synthesis in maxicells of both subunits (Mr = 35,000 (alpha) and Mr = 65,000 (beta) of glycyl-tRNA synthetase. Locations of alpha- and beta-subunit coding regions were established by introduction of Tn5 insertions into various positions within the 5.1-kilobase pair HindIII segment of pTK201 and by determining the effect of each Tn5 insertion on synthesis of alpha- and beta subunits and on enzymatic activity. From the Tn5 insertion analysis, regions encoding the NH2 terminus of the alpha-subunit and of the beta-subunit were approximately defined and these regions were sequenced. To locate rigorously the respective NH2-terminal encoding sections in the DNA sequence, NH2-terminal amino acid sequences of alpha- and beta-subunits were established by standard Edman degradations and these sequences were aligned with the DNA sequence. This analysis established the following: 1) coding regions for the subunits are in tandem; 2) a single promoter is used for transcription of both coding sections and the order of transcription is from alpha to beta; 3) in the 500 nucleotides 5' to the start of the alpha-subunit coding section, there is no sequence arrangement like that found for regulatory regions of bacterial amino acid biosynthetic operons; 4) nine nucleotides serve as the spacer between the TAA stop of the alpha- and the ATG start of the beta-subunit coding regions, thus making both coding regions in the same reading frame; and 5) the TAA stop of the alpha-subunit and the next for nucleotides associated with the intersubunit region are complementary to the 3'-end of 16 S rRNA; this arrangement suggests ribosome re-initiation in the spacer region gives balanced synthesis of both subunits. PMID- 6290470 TI - Antagonistic effects of insulin and beta-adrenergic agonists on the activity of protein phosphatase inhibitor-1 in skeletal muscle of the perfused rat hemicorpus. AB - The extent of phosphorylation of protein phosphatase inhibitor-1 in skeletal muscle rose about 2.5-fold during 60 min of perfusion of the rat hemicorpus preparation and then did not change over the following 30 min. Addition of insulin at 60 min resulted in a 35% fall in inhibitor-1 phosphorylation by 90 min. The rise in inhibitor-1 phosphorylation was due to the presence of catecholamines as evidenced by an accumulation of epinephrine in the perfusate. Removal of the adrenal glands or cannulation of the vena cava prevented the accumulation of epinephrine and the rise in inhibitor-1 phosphorylation. Insulin did not alter the phosphorylation state of inhibitor-1 in the presence of the beta-adrenergic antagonist propranolol where the degree of phosphorylation was low (less than 10%) or at concentrations of isoproterenol (10 nM) where inhibitor 1 was highly phosphorylated (greater than 60%). In preparations with the adrenal glands removed, 0.5 nM isoproterenol produced a 2-fold rise in inhibitor-1 phosphorylation, an effect that was completely prevented by the addition of insulin. The antagonism of 0.5 nM idoproterenol by insulin correlated with a decrease in the muscle content of cyclic AMP. These results suggest that the dephosphorylation of inhibitor-1 may play an important role in the metabolic effects of insulin in vivo. PMID- 6290472 TI - Tityus serrulatus venom contains two classes of toxins. Tityus gamma toxin is a new tool with a very high affinity for studying the Na+ channel. AB - The interaction of TiTx gamma, the major toxin in the venom of the scorpion Tityus serrulatus, with its receptor in excitable membranes was studied with the use of 125I-TiTx gamma. This derivative retains biological activity, and its specific binding to both brain synaptosomes and electroplaque membranes from Electrophorus electricus is characterized by a dissociation constant equal to that of the native toxin-receptor complex, about 2 to 5 pM. This very high affinity results mainly from a very slow rate of dissociation, equivalent to a half-life longer than 10 h at 4 degrees C. There is a 1:1 stoichiometry between TiTx gamma binding and tetrodotoxin binding to the membranes, but neither tetrodotoxin nor any of 7 other neurotoxins that are representative of 4 different classes of effectors of the Na+ channel interfere with TiTx gamma binding. Similarly, local anesthetics and other molecules that affect other types of ionic channels or neurotransmitter receptors have no effect on TiTx gamma binding. However, toxin II from Centruroides suffusus suffusus does compete with TiTx gamma, though its affinity for the receptor is much lower. Since the Centruroides toxin II is known to affect Na+ channel function, these two scorpion toxins must be put into a fifth class of Na+ channel effectors. PMID- 6290473 TI - Studies of the mechanism of cell intoxication by diphtheria toxin fragment A asialoorosomucoid hybrid toxins. Evidence for utilization of an alternative receptor-mediated transport pathway. AB - In order to study the variables important for determining the cytotoxicity of protein-protein hybrid toxins, the disulfide and thioiether conjugates between diphtheria toxin fragment A (DTA) and asialoorosomucoid (ASOR) were synthesized, purified, and tested for cytotoxicity toward isolated rat hepatocytes as monitored by the inhibition of protein synthesis. After 4-h incubation at 37 degrees C, both DTA-ASOR conjugates were nontoxic, but were highly toxic in the presence of an inhibitor of 125I-ASOR degradation. At concentrations inhibiting 125I-ASOR degradation to the same extent, the efficiency of inhibitors to enhance the toxicity of both conjugates varied with the following rank order: colchicine greater than chloroquine greater than NH4Cl greater leupeptin greater than cytochalasin B. In the absence of an inhibitor, both conjugates, after 10-h incubation, also exhibited measurable toxicity at concentrations as low as 10(-9) M, and DTA-S-S-ASOR was 60 times more toxic than DTA-S-ASOR. In a physiological salt solution containing energy source and bovine serum albumin, both conjugates also exhibited partial inhibition of protein synthesis after 4-h incubation and were enhanced by 2 microM colchicine toward complete inhibition with similar C0.5 of 2 X 10(-10) M. The toxicity of the conjugates were prevented in the presence of excess ASOR. These results not only indicate that intracellular degradation is an important factor restricting the toxicity of the conjugates, but also indicate that the toxic entry of DTA activity from these hybrids is independent of lysosomal degradation. Consequently, it appears that conjugate toxicity is dependent on the release of DTA activity from an extralysosomal compartment whose pool size is increased by the inhibitors and the change of medium. The effects of the inhibitors to inhibit 125I-ASOR uptake and degradation were also studied to facilitate interpretation of their enhancement of conjugate toxicity. Specifically, colchicine inhibited both internalization and degradation of the ligand in a saturable fashion with a C0.5 of 0.5 microM and a maximum inhibition of 65%. The data of the present study are interpreted so as to support a hypothesis that two receptor-mediated pathways are involved in the internalization and transport to lysosomes of ASOR and DTA-ASOR conjugates, only one of which is colchicine-sensitive. PMID- 6290474 TI - Proton transport by cytochrome c oxidase from the thermophilic bacterium PS3 reconstituted in liposomes. AB - Cytochrome oxidase from the thermophilic bacterium PS3 which contains three types of polypeptide subunits are reconstituted into liposomes by a freeze-thaw technique. The reconstituted enzyme caused acidification of the medium during cytochrome c oxidation with a stoichiometry of up to 0.8 H+/e. Uptake of K+ ions in the presence of valinomycin occurred with a stoichiometry between 1.5 and 2 K+/e. Dicyclohexylcarbodiimide inhibited the acidification and decreased the stoichiometry of K+ ion uptake to about 1 K+/e. This bacterial oxidase thus appears to be a proton pump with properties similar to the mitochondrial enzyme. PMID- 6290475 TI - Role of carbohydrate in human chorionic gonadotropin. Effect of periodate oxidation and reduction on its in vitro and in vivo biological properties. PMID- 6290476 TI - Characterization of right-side-out membrane vesicles rich in (Na,K)-ATPase and isolated from dog kidney outer medulla. AB - When purified on a sucrose gradient, basolateral membranes from dog kidney outer medulla are found to be very rich in (Na,K)-ATPase; about 50% of the membrane protein is comprised of this enzyme. (Na,K)-ATPase activity is activated 3- to 5 fold by detergent treatment, and this has been previously attributed to the impermeable vesicular nature of the membranes. Porcine trypsin inactivates only that fraction of (Na,K)-ATPase activity seen without detergent, consistent with a right-side-out orientation of membrane vesicles; the trypsin sensitivity and detergent activation of [3H]ouabain binding in the presence of Na+ + Mg2+ + ATP or Mg2+ + Pi are also consistent with this hypothesis. Using nearly isosmotic Hypaque density gradient centrifugation a population of impermeable right-side out membrane vesicles (H1) is separated from a leaky population (H2). (Na,K) ATPase activity in the H1 population is 20-fold activated by detergent and insensitive to porcine trypsin. The vesicle volume is 2.4 microliters/mg, and monovalent cations passively equilibrate with the intravesicular volume on a time scale of 5-30 min. Very rapid ouabain sensitive 22Na efflux from the vesicles is observed when ATP is photolytically released from intravesicular caged ATP. PMID- 6290477 TI - Immunological approaches to the study of membrane receptors. A monoclonal antibody that inhibits the binding of asialoglycoproteins to the rat liver receptor. AB - The major polypeptide (43,000 daltons) of the rat liver receptor for asialoglycoproteins was isolated by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis. Antibodies raised in a goat against this SDS-treated polypeptide exhibited marked cross-reactivity toward the SDS-denatured forms of the two other prominent polypeptides (54,000 and 64,000 daltons) of the receptor preparation. Monoclonal antibodies directed against the receptor were prepared using the spleen cells of mice immunized with the soluble, active receptor purified by affinity chromatography. The most extensively characterized of the monoclonal antibodies, designated D3-5D3, recognized the solubilized receptor and bound to the exterior surface of isolated rat hepatocytes. The binding of D3-5D3 to hepatocytes prevented subsequent binding of the ligand, 125I-asialo orosomucoid. Conversely, occupation of the receptor with ligand inhibited binding of 125I-IgG prepared from D3-5D3 ascites fluid. The secondary structure of the receptor appears to be critical for recognition by D3-5D3, since denaturation of the receptor with 1% SDS, 5% beta-mercaptoethanol at 100 degrees C abolished antibody binding. Under less denaturing conditions (0.1% SDS, 25 degrees C), antigenic reactivity was retained by the receptor. Preparative electrophoresis using the latter conditions permitted the demonstration that D3-5D3 recognized a unique determinant that is present in each of the three polypeptides. PMID- 6290478 TI - Rapid decrease of phosphatidylinositol 4,5-bisphosphate in thrombin-stimulated platelets. AB - The addition of thrombin to horse platelets prelabeled with 32P induces a rapid decrease of the radioactivity from phosphatidylinositol 4,5-bisphosphate. Maximum loss of the radioactivity from phosphatidylinositol 4,5-bisphosphate occurs with 10 s of stimulation and is followed by an increased incorporation of 32P into this lipid. The stimulation of phosphatidylinositol 4,5-bisphosphate loss by thrombin is concentration-dependent. The ionophore A23187, which mobilizes Ca2+, is ineffective in inducing the degradation of phosphatidylinositol 4,5 bisphosphate. Measurements of polyphosphoinositides by phosphorus estimation show that, 10 s after thrombin stimulation, there is a decrease of 15-20% of the total phosphatidylinositol 4,5-bisphosphate without any significant change in phosphatidylinositol 4-monophosphate. It appears that thrombin causes a rapid and transient degradation of phosphatidylinositol 4,5-bisphosphate and that this effect might be related to the initiation of platelet activation. PMID- 6290479 TI - Inhibition by calmodulin of the cAMP-dependent protein kinase activation of phosphorylase kinase. AB - Calmodulin is shown to inhibit both the activation and phosphorylation of phosphorylase kinase by cAMP-dependent protein kinase. Maximal inhibition of both processes was approximately 66% at the highest calmodulin concentration tested (5.5 microM). It was found that the inhibition of phosphorylation was calcium dependent, reversible by trifluoperazine, and specific for the beta subunit of phosphorylase kinase with no significant inhibition of phosphorylation of the alpha subunit. This inhibitory activity of calmodulin appears to be due to an interaction between calmodulin and the substrate, phosphorylase kinase. This finding implies either that the site of exogenous calmodulin interaction with phosphorylase kinase is at the beta subunit or that this interaction results in a conformational change of phosphorylase kinase that inhibits the interaction between cAMP-dependent protein kinase and the beta subunit of phosphorylase kinase. The beta subunit may contain a regulatory site that is recognized by either protein kinase or calmodulin. These findings further substantiate the role of the beta subunits in the activation of phosphorylase kinase and provide an additional example of substrate-directed control of phosphorylation. PMID- 6290480 TI - Newcastle disease virus contains a linkage-specific glycoprotein sialidase. Application to the localization of sialic acid residues in N-linked oligosaccharides of alpha 1-acid glycoprotein. AB - Newcastle disease virus sialidase was found to exhibit strict specificity for hydrolysis of the NeuAc alpha 2 leads to 3Gal linkage contained in glycoprotein oligosaccharides both N-linked to asparagine and O-linked to threonine or serine under conditions that left oligosaccharides containing the NeuAc alpha 2 leads to 2 leads to 6Gal and NeuAc alpha 2 leads to 6GallNAc linkages intact. This was determined, in part, by examining the viral sialidase for its ability to hydrolyze glycoprotein oligosaccharides derivatized with purified sialyltransferases to contain the [14C]NeuAc alpha 2 leads to 3Gal, [14C]NeuAc alpha 2 leads to 6GalNAc, and [14C]NeuAc alpha 2 leads to 6Gal linkages. The viral sialidase was also tested for hydrolysis of the NeuAc alpha 2 leads to 3Gal and NeuAc alpha 2 leads to 6Gal linkages on the N-linked oligosaccharides of alpha 1-acid glycoprotein. Selective hydrolysis of the NeuAc alpha 2 leads to 3Gal linkage was shown by periodate oxidation and by 500-MHz 1H-NMR spectroscopy of native and sialidase-treated glycopeptides. The NMR spectra, together with composition data, further indicated that the NeuAc alpha 2 leads to 3Gal and NeuAc alpha 2 leads to 6Gal linkages were localized to specific branches of the major tri- and tetraantennary oligosaccharides of alpha 1-acid glycoprotein. The results indicate that the Newcastle disease virus sialidase can initiate the selective degradation of N-linked oligosaccharide branches containing the NeuAc alpha 2 leads to 3Gal linkage. PMID- 6290482 TI - In vitro transcription of herpes simplex virus genes. Partial purification and properties of RNA polymerase II from uninfected and infected Hep-2 cells. PMID- 6290481 TI - The cytochrome c peroxidase-catalyzed oxidation of ferrocytochrome c by hydrogen peroxide. Steady state kinetic mechanism. AB - Initial velocities for the cytochrome c peroxidase-catalyzed oxidation of ferrocytochrome c by hydrogen peroxide have been measured as functions of both the ferrocytochrome c (0.27-104 microM) and hydrogen peroxide (0.25-200 microM) concentrations at 25 degrees C, 0.01 M ionic strength, and pH 7 in a cacodylate/KNO3 buffer system Eadie-Hofstee plots of the initial velocity as a function of ferrocytochrome c concentration at constant hydrogen peroxide are nonlinear. A mechanism is proposed which includes random addition of the two substrates to the enzyme and a single catalytically active cytochrome c binding site. The mechanism is consistent with prior studies on cytochrome c peroxidase and fits the steady state kinetic data well. PMID- 6290483 TI - Euglena gracilis chloroplast transfer RNA transcription units. Nucleotide sequence analysis of a tRNATyr-tRNAHis-tRNAMet-tRNATrp-tRNAGlu-tRNAGly gene cluster. AB - A tRNA coding locus in the Bam-Sal 9 region of Euglena gracilis Pringsheim strain Z chloroplast DNA was chosen for detailed study. This DNA contains the previously mapped tRNA coding sequences of the adjacent Euglena chloroplast EcoRI products of EcoV and EcoH (Orozco, E. M., Jr., and Hallick, R. B. (1982) J. Biol. Chem. 257, 3258-3264). The 3.2-kilobase pair Bam-Sal 9 fragment was cloned into the BamHI and SalI cut plasmid vector pBR322, resulting in the recombined plasmid pPG76. The tRNA coding locus was mapped to a region of Bam-Sal 9 that contains portions of both EcoH and EcoV. The DNA sequence of 1-kilobase pair Bam-Sal 9, containing the entire tRNA coding locus, was determined. A cluster of six tRNA genes was found. The gene organization is as follows, where bp is base pair: tRNATyrGUA-64 bp spacer-tRNAHisGUG-14 bp spacer-tRNAMetCAU-4 bp spacer-tRNATrpCCA 27 bp spacer-tRNAGluUUC-6 bp spacer-tRNAGlyUCC. The tRNAMetCAU is believed to be an elongator tRNA. The first four genes are within EcoV. The EcoRI cleavage site that separates EcoV and EcoH is in the tRNAGlu gene. The tRNAGly gene is in EcoH. This is the largest known chloroplast tRNA gene cluster. PMID- 6290485 TI - Low density lipoprotein receptor activity in homozygous familial hypercholesterolemia fibroblasts. AB - We have identified specific low affinity low density lipoprotein (LDL) receptors in skin fibroblasts from two patients previously classified as having LDL receptor-negative homozygous familial hypercholesterolemia (FHC). Km and maximum capacity for cell-associated and degraded 125I-LDL were determined by two independent methods, a traditional technique in which increasing amounts of 125I LDL were added until receptor saturation was achieved and a new technique in which the displacement of a small amount of 125I-LDL tracer was observed during the addition of variable amounts of unlabeled LDL. The Km for specific cell associated 125I-LDL in FHC cells was 3.5-7.3 times that of normal cells and the maximum specific capacity was reduced to 11% of normal. Thus, some FHC cells have reduced affinity as well as reduced capacity for LDL. The FHC cell receptors share many but not all properties of the normal skin fibroblast LDL receptor. Specific degradation of bound 125I-LDL occurred concomitantly with LDL binding and was greatly reduced by the addition of chloroquine, an inhibitor of lysosomal function. Preincubation of FHC cells with cholesterol or LDL resulted in significant suppression of receptor function. Modification of lysine residues of LDL abolished receptor activity in both normal and FHC cells. Treatment of FHC cells with compactin, a cholesterol synthesis inhibitor, resulted in significant increases in specific 125I-LDL binding and degradation compared to FHC cells without compactin treatment. Normal cells also showed increases in 125I-LDL binding and degradation with compactin treatment, but the mean percentage increase in specific 125I-LDL degradation was significantly greater in FHC cells (strain GM 2000, 160 +/- 18%) than in normal cells (29 +/- 8%). PMID- 6290484 TI - Stimulation of mannose-binding activity in the rabbit alveolar macrophage by simple sugars. AB - Mammalian alveolar macrophages are known to bind glycoconjugates with terminal D mannosyl residues (Stahl, P. D., Rodman, J. S. Miller, M. J., and Schlesinger, R. H. (1978) Proc. Natl. Acad. Sci. U. S. A. 75, 1399-1403). Although macromolecules containing D-mannosyl residues, such as bovine serum albumin modified with 2 imino-2-methoxyethyl 1-thio-alpha-D-mannopyranoside (mannose-BSA) (Lee, Y. C., Stowell, C. P., and Krantz, M. J. (1976) Biochemistry 15, 3956-3963), were very potent inhibitors of 125I-mannose-BSA binding to macrophages, the monosaccharides D-mannose, L-fucose, N-acetyl-D-glucosamine, and D-glucose doubled to tripled the binding of 125I-mannose-BSA to intact rabbit lung macrophages at 2 degrees C. The monosaccharide concentration required for maximum stimulation and the extent of stimulation were dependent on the number of D-mannosyl residues attached to the BSA ligand. The lower the number of D-mannosyl residues coupled to BSA, the smaller the effect by D-mannose and the lower the D-mannose concentration at which it occurred. Equilibrium binding analysis indicated that the apparent affinity of cell surface receptor for ligand increased from Kd = 1.0 nM to Kd = 0.3 nM under conditions of maximal stimulation of 125I-mannose43-BSA binding. PMID- 6290486 TI - Energetics and stoichiometry of oxidative phosphorylation from NADH to cytochrome c in isolated rat liver mitochondria. PMID- 6290487 TI - Effects of neighboring DNA homopolymers on the biochemical and physical properties of the Escherichia coli lactose promoter. I. Cloning and characterization studies. AB - To assess the role of neighboring DNA sequences in gene regulation, poly(dA).poly(dT) and poly(dG).poly(dC) were cloned adjacent to promoters of the lactose control region. Recombinant plasmids were constructed which were suitable for large scale purification of restriction fragments containing these promoters, 95-base pair (bp) AluI fragments containing the lack operator and promoter for the lac wild type and for the catabolite gene activating the protein-independent mutant, lac UV5, were cloned into pBR322. Homopolymers of varying lengths were inserted into the -60 region of these promoters using recombinant DNA techniques. Six of the recombinant plasmids were chosen for detailed analysis: wild type (wt); wt-AT, containing 70 bp of poly(dA).poly(dT); wt-GC, containing 23 bp of poly(dG).poly(dC); UV5; UV5-AT, containing 70 bp of poly(dA).poly(dT) and finally UV5-GC, containing 43 bp of poly(dG).poly(dC). These plasmids were characterized by restriction mapping and DNA sequencing. The effects of the DNA homopolymers on the interaction of the Escherichia coli RNA polymerase with the promoters were studied using nitrocellulose filter binding. The results show that poly(dA).poly(dT) increases the level of RNA polymerase binding, whereas poly(dG).poly(dC) has no detectable effect. PMID- 6290488 TI - Effects of Neighboring DNA homopolymers on the biochemical and physical properties of the Escherichia coli lactose promoter. II. In vitro transcription analyses. AB - Transcription studies were conducted on the primary promoter (which initiates at +1) of lac wild type (wt) and UV5 that had insertions of poly(dA).poly(dT) or poly(dG).poly(dC) in their -60 region. This series of recombinant DNAs was designed to study systematically the effect of neighboring DNA sequences on the lac promoter. Investigations using linear templates showed that wild type expression was enhanced by the poly(dA).poly(dT) insertion, whereas UV5 activity was not affected by this polymer. Neither the wt nor the UV5 promoter was influenced by the presence of poly(dG).poly(dC). Quantitation of abortive transcripts from supercoiled templates showed that the influence of the polymers on promoter activity was small. Comparative studies of the primary promoter with a secondary promoter (which initiates near -22), activated by 25% glycerol, were performed. PMID- 6290489 TI - Import of proteins into mitochondria. Cytochrome b2 and cytochrome c peroxidase are located in the intermembrane space of yeast mitochondria. AB - Yeast mitochondria were fractionated into inner membrane, outer membrane, matrix, and intermembrane space. Identity and purity of each fraction were monitored by enzyme assays, dodecyl sulfate-polyacrylamide gel electrophoresis, and immunological detection of characteristic mitochondrial polypeptides. Cytochrome b2 and cytochrome c peroxidase were found to be components of the intermembrane space. The most reliable marker of the outer membrane was a major 29,000-dalton polypeptide component. The availability of submitochondrial fractions provides a basis for studying import of precursor polypeptides into isolated yeast mitochondria. PMID- 6290490 TI - Import of proteins into mitochondria. The precursor of cytochrome c1 is processed in two steps, one of them heme-dependent. AB - The apoprotein of yeast cytochrome c1 is made outside the mitochondria as a larger precursor which is then processed in at least two steps. In the first step, it is transported across both mitochondrial membranes and converted by a matrix-localized protease to an intermediate form whose molecular weight is between that of the precursor and the mature form. The intermediate form is bound to the outer face of the inner membrane. This first step requires an energized mitochondrial inner membrane, but no heme. In the second step, the intermediate form is converted to the mature cytochrome. This second step requires heme; it is blocked in a heme-deficient mutant or in wild type cells treated with an inhibitor of heme synthesis. Import of cytochrome c1 into mitochondria thus proceeds via two distinct heme-free precursors and at least two maturation steps, one of them dependent on heme. PMID- 6290491 TI - Import of proteins into mitochondria. Yeast cells grown in the presence of carbonyl cyanide m-chlorophenylhydrazone accumulate massive amounts of some mitochondrial precursor polypeptides. AB - Cytoplasmically synthesized precursors of mitochondrial polypeptides have previously been observed in trace amounts after pulse labeling of yeast spheroplasts or after in vitro translation of yeast mRNA (Maccecchini, M. L., Rudin, Y., Blobel, G., and Schatz, G. (1979) Proc. Natl. Acad. Sci. U. S. A. 76, 343-347). Some of these precursors are shown here to accumulate in large amounts (up to 150 micrograms/g of cell protein) during growth of a cytoplasmic petite (rho-) mutant in the presence of carbonyl cyanide m-chlorophenylhydrazone, an uncoupler of oxidative phosphorylation. Cytochrome c1 precursor accumulated under these conditions is unstable; it is degraded with a half-life of about 10 min. In contrast, the F1-ATPase beta-subunit precursor is degraded considerably more slowly and, following removal of the uncoupler, can be post-translationally imported into mitochondria where it is processed to the mature polypeptide. PMID- 6290492 TI - Import of proteins into mitochondria. Import and maturation of the mitochondrial intermembrane space enzymes cytochrome b2 and cytochrome c peroxidase in intact yeast cells. AB - The import of cytochrome b2 and cytochrome c peroxidase into mitochondria was investigated by pulse-chase experiments with intact yeast cells combined with subcellular fractionation. Import and processing of the precursors of these intermembrane space proteins is blocked by uncouplers of oxidative phosphorylation, indicating that an "energized" inner membrane is required. Cytochrome b2 is processed in two steps. The first step involves energy-dependent transport across both mitochondrial membranes and cleavage by a matrix-located protease to yield an intermediate which is smaller than the precursor, but larger than the mature protein. The second step involves conversion of the intermediate to the mature form. Whereas the precursor and the mature form are soluble, the intermediate is membrane-bound and exposed to the intermembrane space. The maturation of cytochrome c peroxidase is much slower than that of cytochrome b2. Proteolytic processing rather than import is rate-limiting since cytochrome c peroxidase precursor labeled during a 3-min pulse is already found attached to the outer face of the mitochondrial inner membrane. Import of cytochrome b2 and probably also of cytochrome c peroxidase thus involves energy-dependent transport to the matrix and cleavage by a matrix-localized protease. Maturation of cytochrome b2 proceeds in the sequence: soluble precursor leads to membrane-bound intermediate form leads to soluble mature form. PMID- 6290493 TI - The amino acid sequence of cytochrome c oxidase subunit VI from Saccharomyces cerevisiae. AB - The complete amino acid sequence of the nuclearly coded cytochrome c oxidase subunit VI was determined for a genetically defined haploid strain of Saccharomyces cerevisiae. The subunit contains 108 amino acids, has Mr = 12,627, is acidic (net charge of -9.7 at pH 7) and is quite polar (polarity index, 50.9%). Distribution of charges within the polypeptide chain is highly non random. The NH2- and COOH-terminal regions are predominantly acidic whereas an apolar and a basic region are found in the interior, Subunit VI shows between 28 and 40% sequence homology (depending on the method of alignment) with subunit V of bovine cytochrome c oxidase; since the yeast subunit VI lacks methionine and contains only a single histidine residue very close to the NH2 terminus, it is unlikely that either of the two subunits carries heme alpha in the native enzyme. PMID- 6290494 TI - Glucagon and the Ca2+-linked hormones angiotensin II, norepinephrine, and vasopressin stimulate the phosphorylation of distinct substrates in intact hepatocytes. AB - Recent studies have demonstrated that angiotensin II, catecholamines, and vasopressin can stimulate the phosphorylation of hepatic cytosolic proteins via a Ca2+-linked cyclic AMP-independent mechanism. The present study used high resolution, two-dimensional gel electrophoresis to determine if the proteins phosphorylated in response to the Ca2+-linked hormones were distinct from those affected by glucagon acting via the cyclic AMP-dependent pathway. Intact hepatocytes labeled with [32P]PO4(3-) were stimulated with glucagon, angiotensin II, l-norepinephrine, and vasopressin and over 100 phosphorylated proteins resolved by two-dimensional electrophoresis and autoradiography. Six important enzymes known to be regulated through covalent modification were positively identified, including phosphorylase, phosphofructokinase, pyruvate kinase, fructose-6-phosphate, 2-kinase, phenylalanine hydroxylase, and fructose-1,6 bisphosphatase. Computer analysis of the autoradiograms from control and hormone treated cells demonstrated that glucagon increased the phosphorylation state of 12 phosphoproteins and reduced the phosphorylation of one protein with a Mr = 21,000 and a pI = 5.9. The Ca2+-linked hormones stimulated the phosphorylation of 7 phosphoproteins and also reduced the phosphorylation state of the 21,000-dalton protein. Angiotensin II, l-norepinephrine, and vasopressin had equivalent effects on protein phosphorylation. There were six protein substrates uniquely affected by glucagon and one phosphoprotein uniquely stimulated by the Ca2+-linked hormones. Seven substrates were affected by stimulation of the cell with either glucagon or the Ca2+-linked hormones. These results demonstrate that, while there is overlap in the substrates affected by glucagon and the Ca2+-linked hormones, each pathway is able to affect the phosphorylation of unique substrates. This finding suggests that the two types of hormones may have some distinct effects on hepatic function.U PMID- 6290495 TI - Immunoblot analysis of low density lipoprotein receptors in fibroblasts from subjects with familial hypercholesterolemia. AB - This paper describes a sensitive method for study of the isoelectric point and molecular weight of immunoreactive low density lipoprotein (LDL) receptors of cultured human fibroblasts. The fibroblast receptors are solubilized with Triton X-100, partially purified by batch elution from DEAE-cellulose, and subjected to two-dimensional isoelectric focusing/sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The proteins are transferred electrophoretically to nitrocellulose paper which is then incubated with a mouse monoclonal antibody (IgG-C7) directed against the LDL receptor, followed by an 125I-labeled antibody against mouse IgG. The receptor-bound monoclonal antibody is localized by autoradiography. By this technique, the immunodetectable LDL receptors from normal human fibroblasts migrate as a single spot with an isoelectric point of 4.3 and a Mr of approximately 160,000. In one patient with homozygous familial hypercholesterolemia whose cells fail to bind 125I-labeled IgG-C7, no immunoreactive LDL receptor spot was detected after electrophoresis. We also studied LDL receptors from three homozygotes whose cells bind 125I-IgG-C7, i.e. cross-reacting material-positive mutants. Their immunodetectable receptors were indistinguishable from normal receptors in terms of isoelectric point and molecular weight. Similarly, the receptors from one patient with the internalization-defective form of familial hypercholesterolemia showed normal electrophoretic migration. The immunoblotting technique should prove useful in analyzing structural alterations, if they exist, in LDL receptors from other subjects with cross-reacting material-positive forms of familial hypercholesterolemia. PMID- 6290496 TI - Isolation and physical mapping of nitrogen fixation genes from the cyanobacterium Anabaena 7120. PMID- 6290497 TI - The elution of foot-and-mouth disease virus from vaccines adjuvanted with aluminium hydroxide and with saponin. PMID- 6290498 TI - Factors affecting the reproducibility of the serum neutralization test for infectious bursal disease. PMID- 6290499 TI - The effect of initial temperature on free radical decay in PMMA bone cement. AB - The effect of varying the initial temperature of the components of poly(methyl methacrylate) (PMMA) bone cement has been investigated. Electron paramagnetic resonance (EPR) spectroscopy was used to monitor free radical decay during the curing of the cement. Samples cured in saline at 37 degrees C were found to exhibit first-order decay kinetics for the polymerization radicals for approximately one week after mixing. This indicates that the decay did not take place by combination or disproportionation and was probably due to a transfer process. Decreasing the temperature of the bone cement components prior to mixing resulted in smaller decay rates, but still with first-order kinetics. This decrease in decay rate with lower initial temperature may be due to decreased porosity of the cement, possibly due to decreased monomer evaporation. SEM micrographs of the samples were consistent with this change in porosity. PMID- 6290501 TI - The tumor-protective effect of selenium in an experimental model. AB - The possibility of a tumor-protective effect of selenium on the growth of benzpyrene-induced sarcoma was studied in an assay with Balb/c mice. The animals received 4 parts/10(6) selenium in their drinking water for 12 months prior to subcutaneous injection of benzpyrene to induce sarcomas. In contrast to unpretreated controls, the selenium-exposed animals developed significantly less and smaller tumors in a given time. These results are compared to those of other authors. Several mechanisms for selenium influence on benzpyrene metabolism are discussed. PMID- 6290500 TI - Hemopoietic stem cells and target cells in murine-virus-induced leukemias. PMID- 6290502 TI - Alteration of binding properties and cytoskeletal attachment of nerve growth factor receptors in PC12 cells by wheat germ agglutinin. AB - Incubation of PC12 cells preloaded with 125I-nerve growth factor (NGF) reveals rapidly and slowly dissociating binding components indicative of a heterogeneous population of receptors. If the cells are previously exposed to wheat germ agglutinin (WGA) for 30 min, NGF now binds to an apparently homogeneous receptor population which exhibit slow dissociation kinetics. Total binding is also reduced by 50%. If WGA is added subsequent to 125I-NGF, total binding is not diminished, but rapidly dissociating receptors occupied with NGF are all converted to the slowly dissociating form. This conversion of receptors occurs rapidly, reaching completion within 2 min at 37 degrees or 4 degrees C, and is unaffected by metabolic energy poisons, suggesting that WGA-induced slowly dissociating receptors are not the product of internalization. The effects of the lectin are blocked by the sugar N-acetyl-D-glucosamine, and the lectin-induced slowly dissociating receptors are converted back to rapidly dissociating receptors by addition of this same sugar. WGA also affects the association of the NGF receptor with the Triton X-100 cytoskeleton. Greater than 90% of bound 125I NGF becomes associated with Triton X-100 insoluble cytoskeletons in the presence of the lectin, compared with less than 20% before lectin addition. Cytoskeleton association of the NGF receptor by WGA shows similar kinetics as the conversion of rapidly to slowly dissociating receptors. This interaction may be involved in the alteration of NGF-receptor binding properties produced by this lectin. PMID- 6290503 TI - Dual pathways for epidermal growth factor processing after receptor-mediated endocytosis. AB - The binding, internalization, intracellular translocation, and degradation of epidermal growth factor (EGF) were studied in mouse Swiss/3T3 fibroblasts under two different physiological conditions at 37 degrees C. In serum-containing medium the maximal level of cell-bound EGF was maintained for at least 8 h without appreciable degradation in contrast to serum-free conditions. These phenomena were correlated with a difference in the intracellular site to which the receptor-bound EGF was delivered as studied using Percoll density gradients. In serum-containing medium the majority of cell-bound EGF was initially taken up into a Golgi-like vesicle of density 1.046, corresponding to the marker galactosyl transferase, and then delivered to a population of vesicles with similar density as lysosomes ( = 1.068-1.110). A portion of the EGF became degraded and was released from the cell into the medium while the remainder stayed with the cells, intact, for a long period of time. In serum-free medium, EGF became associated with a heterogeneous population of vesicles with a mean density of 1.050 which do not correspond to any of the marker enzymes for subcellular organelles for which we have tested (Golgi, endoplasmic reticulum, plasma membrane, lysosomes). It is then transferred to lysosome-like vesicles ( = 1.068-1.110). We therefore propose that EGF is processed through two separate endocytotic routes which are regulated by the cell depending upon its physiological state. PMID- 6290506 TI - Nuclear ligation of RNA polynucleotide kinase products. AB - RNA polynucleotide kinase has been shown to transfer [gamma 32P] from ATP to 5-OH termini of endogenous nuclear RNA. The products of this reaction have been isolated in RNA larger than 125 after in vitro incubation of mouse L cell nuclei. About 20%-30% of these 5'-OH kinase products are polyadenylated. A sizeable fraction of the [gamma 32P] label from ATP is also found in internal phosphodiester bonds after 30-minute nuclear incubation in vitro. The possibility of substantial [32P] 1) 2mM nucleoside triphosphates in the incubation medium, 2) limited nearest-neighbor distribution 3' and 5' to the phosphodiester bond compared with that from [alpha 32P] UTP, 3) different nearest-neighbor distribution for RNA molecules greater than 125 and 12-3S, 4) relative insensitivity of the [gamma 32P] incorporation to alpha-amanitin as compared with total RNA synthesis, 5) internal [32P] appearance in RNA greater than 12S in less than five minutes of incubation, and 6) less than 0.03% to 0.6% of the total [32P] in the alpha position of nucleoside triphosphates after 30 minutes of incubation. The [gamma 32P] incorporation was dependent on high ATP concentration and was insensitive to competition by inorganic phosphate. These results are consistent with the levels of 5' RNA polynucleotide kinase activity in L cell nuclei and suggest the presence of an RNA ligase that can utilize the termini generated by the 5'-OH RNA kinase in a ligation reaction. PMID- 6290504 TI - Electrophysiological and pharmacological properties of cultured rat thyroid cells. AB - The electrophysiological properties of a hormone-dependent, differentiated thyroid epithelial cell strain were studied using intracellular microelectrodes. The average membrane potential of solitary, isolated cells was -78.4 +/- 1.3 mV. The membrane potential depolarized 55 mV per tenfold increase in extracellular potassium concentration. Weak electrical coupling was recorded between contiguous cells. Like thyroid cells in vivo, these cells did not generate action potentials. In some cells a spontaneous, slow transition in the membrane potential from -80mV to -30 mV was accompanied by an increase in input resistance. Membrane potential transitions could be induced by perfusing cells with isotonic Hanks solutions saturated with CO2 (pH = 5.5) or by perfusing cells with hypotonic Hanks solutions (190-290 mOsm/kg). Membrane potential transitions were due to a decreased potassium permeability. Noradrenaline elicited both a fast depolarization and a slow depolarization. The fast depolarization was due to an increase in conductance of Na+ channels and of Cl- channels. Intracellular injection of Ca++ elicited the fast depolarization. Intracellular injection of EGTA or cobalt abolished the fast depolarization. Replacement of extracellular Ca++ by Mg++ did not affect the fast depolarization. Thus, the fast depolarization was due to accumulation of intracellular Ca++. The fast depolarization was abolished by the alpha adrenergic blocker phentolamine (10(-6) M), and was not abolished by the beta adrenergic blocker propranolol (10(-5) M). PMID- 6290505 TI - Murine cell surface transferrin receptor: studies with an anti-receptor monoclonal antibody. AB - A rat monoclonal antibody against the murine transferrin receptor has been identified. The receptor is a 95,000 molecular weight species that exists in the cell membrane as a disulphide-bonded dimer. Whereas 29 of 29 murine hematopoietic tumor cell lines express detectable numbers of transferrin receptors, less than 1% of adult thymocytes or spleen cells and only 5% of bone marrow cells are positive. However, fetal liver and neonatal spleen contain substantial numbers of transferrin receptor-positive cells. Induction of Friend cells in vitro with dimethyl-sulphoxide leads to an overall increase in the expression of transferrin receptors on the cell surface. The anti-transferrin receptor antibody we have obtained partially blocks iron uptake from 59Fe-transferrin by a variety of murine cell lines and inhibits the growth of a murine myeloma cell line in vitro. PMID- 6290507 TI - Down regulation and recovery of the epidermal growth factor receptor in serum supplemented versus defined medium. AB - The down regulation of surface membrane receptors for 125I) epidermal growth factor (EGF) has been evaluated in normal and SV40-transformed human fibroblasts (WI38) under conditions of serum-supplemented versus defined growth media. Both normal and transformed WI38 cells down regulate and recover the EGF receptor and these processes do not differ significantly in serum-supplemented versus defined media. These data are in contrast to a recent study that reported that the HeLa cell does not down regulate the EGF receptor in defined media, whereas it does in serum-supplemented media. PMID- 6290508 TI - The amount of a specific cellular protein (p53) is a correlate of differentiation in embryonal carcinoma cells. AB - A specific cellular protein of molecular weight of 53-55,000 (p53) has been shown to be induced in all SV40 transformed cells. A similar protein has also been shown to be present in embryonal carcinoma cells and in midgestation murine embryo primary cells, which are not infected by SV40. In embryo cell primaries the amount of the protein was shown to decrease with the increase in the stage of embryo development. As differentiation or decrease in cell growth rate can account for this, and since the growth rate of embryo primary cells cannot be measured, we chose to investigate various embryonal carcinoma cells. We report that the p53 is present in a pluripotent embryonal carcinoma cell OTT6050, and in its differentiated parietal endoderm derivative, PYS-2 cells. The amount of p53 is higher in the undifferentiated EC stem cells than in the differentiated PYS-2 (parietal endoderm) cells. The amount of the protein decreases in F9 embryonal carcinoma cells induced to differentiate to a parietal endoderm cell type by treatment with retinoic acid, as it does following spontaneous differentiation of OTT6050 EC cells. To determine if a change in growth rate, rather than differentiation, might account for the diminished levels of this protein, the amount of p53 was measured in growing and in growth arrested cell populations. When the growth rate of F9 cells was reduced by treatment with 8-bromocyclic AMP there was no change in the amount of p53. The half life of the p53 was compared in the undifferentiated and the differentiated cell types to determine if a change in stability might account, in part, for the altered levels of this protein. The p53 is found to be most stable in the SV40 transformed established clonal cells. It is less stable in the fibroblast clonal cells which were not transformed by SV40. The results of these experiments indicate that a decrease in the amount of p53 primarily correlates with differentiation in the embryonal carcinoma cell lines studied and not with cell growth rate. Furthermore, the decrease appears to be related (in part) to the decreased stability of the p53. PMID- 6290509 TI - Oxygen-stimulated cytochrome oxidase assembly in hepatocyte monolayer cultures. PMID- 6290510 TI - Regulation of Na+,K+ pump activity by nerve growth factor in chick embryo dorsal root ganglion cells. AB - Nerve growth factor (NGF) is required for the growth and development of sensory and sympathetic neurons. Incubation of chick dorsal root ganglionic cells without NGF resulted in a decrease of active (Na+,K+-pump-mediated) K+ influx over a period of several hours. Addition of NGF to NGF-deprived cells caused 1) a return of the active K+ influx to the values occurring in cells continuously exposed to NGF, preceded by 2) a very rapid, but transient overstimulation of the Na+,K+ pump-mediated K+ influx. Restoration of normal Na+,K+-pump activity occurred at NGF concentrations of 1 biological unit/ml or greater, whereas the NGF concentration in the 1-100 biological unit/ml range affected the rapidity with which the pump restoration took place. The transient pump behavior was only observed in NGF-deprived cells and could not be elicited in NGF-supported steady state cells or in cells having already received delayed NGF once. This transient Na+,K+-pump behavior was exclusively displayed in conjunction with a high intracellular Na+ concentration. Decreasing the external Na+ concentration below 70 mM reduced the hyperstimulation response to NGF, until at 10 mM Na+ the delayed presentation of NGF caused no overshoot at all. The effect of NGF on the Na+,K+-pump was specific for the NGF molecule and could not be mimicked by other proteins. PMID- 6290511 TI - Temperature-dependent alteration of cellular morphology by cholera toxin in rat liver epithelial cells which are ts for maintenance of transformed properties. AB - Cholera toxin via its ability to increase intracellular cyclic AMP levels can induce drastic changes in cell morphology. This report describes a temperature sensitive mutant of chemically transformed rat liver epithelial cells which only display cell shape alterations in response to cholera toxin at the permissive temperature. Shift up-shift down experiments indicate that the change in the response occurs fairly rapidly, i.e., within 2 hours at the new temperature. The behavior of the temperature sensitive cells at the nonpermissive temperature mimics that of the untransformed rat liver epithelial cells (i.e., no morphological change in response to cholera toxin) while at the permissive temperature the positive cell shape change is identical to that exhibited by chemically transformed rat liver epithelial cells. The temperature sensitive response to cholera toxin is not a function of cyclic AMP production, since the amount of cyclic AMP found as a function of either time or concentration of cholera toxin is quite similar in cells treated at either temperature. PMID- 6290513 TI - Regulation of amino acid transport in L6 myoblasts. II. Different chemical properties of transport after amino acid deprivation. AB - The mechanism of stimulation of amino acid transport system A caused by amino acid deprivation in L6 cells was investigated. In cells loaded with alpha aminoisobutyric acid (AIB), amino acid deprivation increased the rate of proline uptake only after the intracellular [AIB] dropped below 7 mM. Efflux of proline was not sensitive to the presence of proline in the outer medium (with or without external Na+), suggesting that efflux through system A (and possibly uptake) is not susceptible to transinhibition. Transport (stimulated uptake) into amino acid deprived cells and that into amino acid-supplemented cells differed in several chemical properties: 1) In the former group, transport was higher at lower pH values than in the latter, and the optimum pH values were 7.5 and 7.8, respectively. 2) Unlike proline uptake in supplemented cells, uptake in deprived cells was inhibited by 50% with N-ethylmaleimide (1 mM) or by 50 microM p chloromercuribenzoate (PCMBS). Inhibition by PCMBS was not due to collapse of the Na+ gradient. The mercurial inhibited only the deprivation-induced stimulation of transport, bringing the rate of proline uptake to the "basal" uptake level observed in amino acid-supplemented cells. Proline uptake was not stimulated by a second deprivation following treatment with PCMBS and a supplementation deprivation cycle. However, in untreated cells, or by reversing mercaptide formation with dithiotreitol, the second deprivation stimulated transport. Deprivation at 4 degrees C did not elicit stimulation of proline uptake. Cycloheximide prevented the stimulation and decreased the rate of proline uptake in deprived cells more efficiently than in supplemented cells. Actinomycin D prevented stimulation when added at the onset of deprivation. The above data indicate that stimulation of transport by deprivation is protein synthesis dependent and that the stimulated transport had chemical properties distinct from the "basal" transport in supplemented cells. The evidence presented is consistent with a model of activation of a finite pool of transporters upon deprivation, the chemical characteristics of which differ from those of the "basal" transport system. PMID- 6290512 TI - Regulation of transferrin receptor expression in concanavalin A stimulated and Gross virus transformed rat lymphoblasts. AB - Expression of the cell surface receptor for the serum glycoprotein transferrin has been correlated with cellular proliferation in normal lymphocytes undergoing mitogen or antigen induced proliferative responses. In the present study, the expression of transferrin receptor in Concanavalin A stimulated rat lymphocytes or Gross virus transformed lymphoma cells has been examined with respect to the following questions: (1) is expression of receptor activity related to blastogenesis or to the subsequent IL-2 dependent DNA synthetic activity, and (2) is transferrin receptor expression regulated in similar fashion in both normal and malignant lymphoblasts? Scatchard analysis of saturation binding data illustrated that binding site number increased and subsequently decreased during the response while the receptor affinity for transferrin remained constant. These findings were confirmed by SDS-polyacrylamide gel electrophoretic analysis of radiolabeled cell surface proteins which specifically interact with transferrin. Examination of nonproliferating normal lymphoblasts (96 hr post Con A stimulation) compared with the same population of cells stimulated to reinitiate DNA Synthesis with a partially purified preparation of Interleukin 2 (IL-2) showed that transferrin receptor expression was tightly linked to the IL-2 dependent stimulation of DNA replication. This coordinate regulation of receptor expression was markedly less stringent in retrovirus transformed thymic lymphoma cells. PMID- 6290514 TI - The role of calcium in the initiation of superoxide release from alveolar macrophages. AB - The role of calcium in the release of superoxide anion (O2-) was examined in alveolar macrophages after stimulation with the soluble stimuli: concanavalin A (Con A), N-formyl methionyl phenylalanine (FMP), and the calcium ionophore. A23187. The release of O2- by Con A was unaffected over a wide range of extracellular calcium concentrations (20 microM to 3 mM), whereas increasing the extracellular calcium above 2 mM inhibited FMP-stimulated O2- release. In contrast, A23187 did not stimulate O2- release in calcium-free medium (less than or equal to 30 microM). The addition of EGTA (50 microM) to calcium-free medium had no effect on Con A stimulation of O2- release or FMP-stimulated O2- release. These results suggest that, for the three soluble stimuli, there are different roles for Ca+2 in the activation and transmission of stimulatory signals across the cell membrane. Con A- or FMP-stimulated calcium efflux from calcium-loaded cells in either calcium-free medium or 0.5 mM calcium-containing medium. In calcium-free medium, FMP transiently retarded 45Ca+2 uptake, while in 0.5 mM calcium-containing medium, FMP transiently stimulated 45Ca+2 uptake. For either Con A or FMP, calcium efflux preceded O2- release by 30-45 sec. Quinine, an agent that blocks membrane hyperpolarization in macrophages, completely blocked O2- release by concanavalin A or FMP and inhibited 45CA+2 efflux by 50% or more for both agents. These results support the hypothesis that redistribution of cellular Ca+2 is one of the initial steps leading to the release of O2-. PMID- 6290515 TI - Glass capillary or fused-silica gas chromatography--mass spectrometry of several monosaccharides and related sugars: improved resolution. AB - The gas chromatographic separation of several monosaccharides and related sugars derivatized by methoxylation and trimethylsilylation reactions was optimized with glass capillary (SP-2250) and fused silica (SP-2100) columns. Individual sugars included aldoses, ketoses, polyols, acidic forms and N-acetylated amino sugars. Peaks were detected by selected ion monitoring (SIM). The fused silica column gave complete resolution of all peaks (two per hexose and one per hexitol) arising from glucose, galactose, mannose, fructose, sorbitol, mannitol and dulcitol. The resolution of these sugars with the glass capillary column was not as good, but full differentiation was possible on the basis of SIM. Because the fused silica column gave a better resolution of 33 sugars tested and was more easily installed than the glass capillary column, it was utilized for quantitative analysis. A deuterated algal sugar mixture used for quantitation by isotope dilution was found to contain glucose, galactose, mannose, xylose, arabinose, ribose and rhamnose. Full recoveries were obtained of various amounts of glucose, galactose, mannose, fructose and xylose added to human serum. PMID- 6290516 TI - Simultaneous determination of thioridazine and its S-oxidized and N-demethylated metabolites using high-performance liquid-chromatography on radially compressed silica. AB - A method for simultaneously quantifying thioridazine, northioridazine, thioridazine-2-sulfoxide, thioridazine-2-sulfone and thioridazine-5-oxide in serum and plasma is described. Following solvent extraction these compounds were separated by high-performance liquid chromatography on radially compressed silica gel and detected by UV absorbance at 254 nm. Chromatography time is less than 7 min. The relative retention of these compounds as a function of the methanol and methylamine content of the mobile phase is discussed. Practical limits of detection, based upon on assayed plasma or serum volume of 1 ml, were 20 ng/ml for thioridazine-5-oxide and 10 ng/ml for the other compounds. The coefficient of variation for all compounds was less than 13%. The method is compared with more conventional high-performance liqiud chromatographic and gas chromatographic methodology. PMID- 6290517 TI - Determination of impurities in propranolol hydrochloride by high-performance liquid chromatography on dynamically modified silica. AB - A rapid high-performance liquid chromatographic method has been elaborated for the separation and determination of small amounts of impurities in propranolol hydrochloride. The separation was achieved on a column of bare silica (Zorbax SIL) with methanol-water-0.2 M phosphate buffer pH 8.0 (70:25:5) containing 2.5 mM of cetyltrimethylammonium (CTMA) bromide as the eluent. The concentrations of methanol and CTMA as well as the pH of the phosphate buffer were found greatly to affect the separation. The selectivity of the system towards a mixture of propranolol and three possible impurities was investigated using different brands of silica. Only minor variations were found relative to those of a chromatographic system based on chemically bonded ODS silicas from the same sources. The method is also suitable for identification purposes, being able to separate most beta-blocking drugs of structures similar to that of propranolol. PMID- 6290518 TI - Simultaneous assay of cyclic AMP and cyclic GMP phosphodiesterase activity by anion-exchange column chromatography. PMID- 6290519 TI - Purification of retrovirus genomic RNA suitable for chemical radioiodination. AB - An efficient method for the purification of genomic RNA from the retrovirus, caprine arthritis-encephalitis virus, is described. The method utilizes proteinase K, extraction with sodium perchlorate and chromatography on oligo(dT) cellulose and results in highly purified RNA capable of being chemically iodinated with Na125 I to high specific radioactivity. The iodinated RNA exhibits 80-90% precipitability in 5% trichloroacetic acid and is greater than or equal to 99% sensitive to hydrolysis by ribonuclease. Several alternative methods which are effective for the preparation of eukaryotic ribosomal RNA are unreliable for purification of retrovirus RNA suitable for radioiodination. PMID- 6290520 TI - A method for the purification of rotaviruses and adenoviruses from faeces. AB - Rotaviruses and non-cultivable enteric adenoviruses were purified from large volumes of faecal extracts using polyethylene glycol precipitation, centrifugation through 45% w/v sucrose and segregation by density in a cesium chloride density gradient. Yields were high and the preparations were pure enough to use in the production of type-specific antisera and in viral polypeptide analysis. Human rotavirus particles of density 1.38 g/ml were shown to be non infectious for LLC-MK2 tissue culture cells. PMID- 6290521 TI - Inhibitory influence of exogenous oxytocin on adrenocorticotropin secretion in normal human subjects. AB - Oxytocin has been suggested to have behavioral effects opposite to those of vasopressin, and exogenous vasopressin is known to induce ACTH release in man. Thus, we tested the influence of exogenous oxytocin on blood levels of ACTH and cortisol during insulin-induced hypoglycemia and after vasopressin injection. Our results demonstrate an inhibitory effect of exogenous oxytocin on ACTH release and support the hypothesis of a reciprocal, balanced modulation of behavioral and neuroendocrine function by the two closely related neurohormones, vasopressin and oxytocin. PMID- 6290522 TI - Effects of synthetic corticotropin-releasing factor and dopamine on the release of immunoreactive beta-endorphin/beta-lipotropin and alpha-melanocyte-stimulating hormone from human fetal pituitaries in vitro. AB - The effects of synthetic corticotropin-releasing factor (CRF) and dopamine on immunoreactive beta-endorphin/beta-lipotropin (i beta-END/LPH) and alpha MSH release were studied in superfused human fetal pituitary glands. CRF (20 ng) stimulated the release of i beta-END/LPH in four anterior hemipituitaries from fetuses older than 20 weeks in gestation. There was no effect on three anterior hemipituitaries from fetuses of 19-20 weeks gestation. CRF had no effect on i beta-END/LPH or alpha MSH secretion from neurointermediate lobes regardless of fetal age. Dopamine (10(-6) M) had no effect on i beta-END/LPH or alpha MSH secretion from either anterior or neurointermediate lobes. The data suggest that anterior pituitary responsiveness to CRF develops at about 20 weeks gestation and that fetal neurointermediate lobe secretion of peptides is not regulated by CRF. PMID- 6290523 TI - Studies on primate gonadotropin receptors: characterization of the rhesus monkey testicular follicle-stimulating hormone receptors. AB - The properties of adult rhesus monkey testicular FSH receptor was investigated in these experiments. The interaction of 125I-labeled human FSH with a monkey testicular particulate fraction is a time- and temperature dependent phenomenon. Equilibrium of hormone-receptor interaction occurred by about 4-6 at 37 or 34 C, was slow at 25 C, and was extremely slow at 4 C. Maximum binding occurred at pH 7 7.5, with a requirement of 5-10 mM MgCl2 or CaCl2. The half-life of the receptor with exposed sites for hormone interaction was temperature related (1 h at 37 C, 1.5 h at 34 C, 6 h at 25 C, and 36 h at 4 C). Occupancy of these sites by the labeled hormone rendered the receptor more stable. The hormone-receptor complex was highly stable, as shown by the fact that excess unlabeled hormone was unable to displace the already bound labeled hormone from the receptor. Conditions unfavorable for hormone-receptor interaction, such as pH 5.0 or pH 10 or high salt concentration (0.5 M MgCl2), induced the maximum dissociation of the preformed hormone-receptor complex. The primate testis FSH receptor was inactivated by trypsin, phospholipase C, and reducing agents, but it was not influenced by nucleases. Neuraminidase treatment of the particulate receptor may have enhanced its ability to bind labeled human FSH. PMID- 6290524 TI - Adrenocorticotropin and cortisol responsiveness to thyrotropin-releasing hormone and luteinizing hormone-releasing hormone discloses two subsets of patients with Cushing's disease. AB - In 23 consecutive patients with Cushing's disease and 52 control subjects, the responses of ACTH and cortisol to TRH and LRH were investigated. From the pattern of cortisol levels after the administration of the releasing hormone in the controls, a criterion for paradoxical responsiveness could be derived (maximum cortisol increase, greater than 6.0 micrograms/100 ml). According to this criterion, 9 patients with Cushing's disease showed a paradoxical responsiveness to one or both releasing hormones (3 to both TRH and LRH, 3 to TRH alone, and 3 to LRH alone; group I). In all patients tested, paradoxical responses of cortisol were preceded by paradoxical increments in ACTH. The remaining 15 patients showed no paradoxical increments in ACTH or cortisol after TRH or LRH (group II). ACTH levels in group I (89 +/- 28 pg/ml) were significantly lower than those in group II (185 +/- 164 pg/ml; P less than 0.02). Nevertheless, in both groups, a similar plasma cortisol level was found, suggesting a relatively higher bioactivity of ACTH in group I. A second difference between both groups was a lower amplitude of cortisol variability during the day in group I. The 2 groups did not differ in clinical data, such as age, sex distribution, sellar volume, and duration of disease, or dexamethasone suppressibility, bromocriptine sensitivity, and basal PRL levels. These latter findings do not favor an intermediate lobe origin of Cushing's disease in patients with paradoxical responses to TRH/LRH. To conclude, TRH/LRH responsiveness of ACTH/cortisol discloses two subsets of patients with Cushing's disease. PMID- 6290526 TI - Monoclonal antibodies to human T cell subsets: use for immunological monitoring and immunosuppression in renal transplantation. AB - Sequential monitoring of peripheral blood T cell populations was performed in normal control individuals and in 55 renal allograft recipients. In addition, 66 patients exhibiting prolonged renal allograft survival were each evaluated at least once with the same monoclonal antibodies. A normal or elevated OKT4:OKT8 ratio, especially in the presence of rising numbers of OKT4+ cells was predictive of possible future graft rejection. This observation, when followed by rising serum creatinine values coupled with biopsy evidence of glomerulopathy, was found in at least 15% of graft recipients. In a small number of patients, graft rejection episodes were successfully suppressed by administration of therapeutic doses of OKT3 antibody. However, production of anti-OKT3 antibodies was observed and may be expected to limit administration of this agent to a single short course in each patient. PMID- 6290525 TI - Altered fractional tetrahydroaldosterone excretion during pharmacological blockade and activation of the renin-aldosterone system. AB - Tetrahydroaldosterone (THA) is the principal metabolite and generally a good index of aldosterone secretion. This study undertakes the evaluation of the aldosterone secretion rate (ASR) and excretion of THA during pharmacological blockade and activation of the renin-aldosterone system. THA was measured by a simplified RIA and compared to ASR over a period of 28 days in 18 normotensive volunteers receiving either 1) a diuretic [hydrochlorthiazide (HCTZ), 50 mg/day], 2) an angiotensin-converting enzyme inhibitor (MK-421, 10 mg/day), or 3) combined therapy [HCTZ (50 md/day) plus MK-421 (10 mg/day)] in a metabolic unit on a controlled diet. Results of this study at 28 days indicate that 1) HCTZ, while producing secondary hyperaldosteronism, lowered the fractional THA excretion (defined as THA/ASR) (from 0.51 to 0.33; P less than 0.05); 2) MK-421 produced hypoaldosteronism and a slight increase in the THA/ASR (from 0.43 to 0.53; 0.05 less than P less than 0.01); 3) combined HCTZ and MK-421 resulted in a normalization of both aldosterone secretion and THA/ASR (from 0.51 to 0.50). In conclusion, HCTZ decreases the THA/ASR whereas MK-421 tends to increase it. Combined administration of HCTZ and MK-421 restores the THS/ASR to normal. Therefore, the determination of THA excretion can be an inaccurate index of aldosterone secretion when measured during either pharmacological blockade or activation of the renin-aldosterone system. PMID- 6290527 TI - OKT3 incubation of donor marrow for prophylaxis of acute graft-vs.-host disease (GvHD) in allogeneic bone marrow transplantation. AB - Following allogeneic (matched or minor mismatched) bone marrow transplantation with marrow incubated with OKT3, 17 patients have been observed for at least 60 days for the development of acute graft-vs.-host disease. One milligram of the monoclonal (murine) antibody OKT3 was incubated with the donor marrow for one hr at room temperature prior to infusion. It was anticipated that coated/opsonized immune competent T lymphocytes would subsequently be phagocytosed by the recipient reticulo-endothelial system. Three of 17 patients developed Grade II or greater acute graft-vs.-host disease. In two, this proved fatal when complicated by cytomegalovirus infection. Eleven patients showed no evidence of acute graft vs.-host disease and three had transient limited skin rashes (Grade I graft-vs. host disease). Opsonization of T lymphocytes has reduced the incidence of severe acute graft-vs.-host disease in this unit from 79% (historical group) to 18% when added to prophylactic methotrexate given according to the Seattle protocol. PMID- 6290528 TI - T cell subset abnormalities in tissue lesions developing during autoimmune disorders, viral infection, and graft-vs.-host disease. AB - The authors review a large body of contemporary immunohistologic findings on the tissue distribution of T lymphocytes in normal and pathological conditions. The suggestions for technological advances in this field are: signal amplification using mixtures of monoclonal antibodies directed against different epitopes on the same antigen (e.g. OKT4A+B+D), triple layer amplification systems using hapten-labelled antibodies, and informative double staining methods with combinations of antibodies labelled with different fluorochromes or enzymes. Review of histological observations in a series of human diseases suggests that imbalances of OKT4+ and OKT8+ subsets of T lymphocytes may represent different types of immunoregulatory disorders. Rheumatoid arthritis and sarcoidosis appear to involve a high level of OKT4+ subpopulation response coupled with an associated appearance of a special type of HLA-DR+ macrophages. It remains to be seen whether normal or self-limited immunological responses (early stages of bacterial infection or delayed-type hypersensitivity reactions) produce OKT4+ and macrophage responses that are characteristically different. Meanwhile, excessive levels of OKT8+ cells have been found in a wide range of recognized or presumed immunoregulatory disorders including: graft-vs.-host reaction and viral infections. These disorders, as well as primary biliary cirrhosis and lichen planus, appear to possess both overlapping and disparate clinical characteristics, and the immunohistological observations may reflect the functional heterogeneity of OKT8+ populations in these diseases. These studies show that histologically meaningful heterogeneity can already be demonstrated for the OKT8+ lymphocyte group. PMID- 6290529 TI - The effects of immunomodulation on peripheral T cell subsets. AB - Monoclonal antibody assays were employed to monitor modifications induced in human peripheral lymphocyte subsets by thymectomy or the administration of a series of immunomodulating drugs: synthetic thymic factor, cimetidine or various combinations of anti-thymocyte globulin, azathioprine and steroids. In patients with myasthenia gravis, thymectomy produced a gradual progressive decrease in the elevated OKT4/OKT8 ratios associated with this disease until normal ratios were achieved after one year. Administration of synthetic thymic factor to three immunodeficient children for one month produced increased serum IgA levels accompanied by a normalization of proportions of total T cells and T cell subsets. Four of five uremic patients receiving cimetidine exhibited a marked increase in the percentage of OKT8+ T cells observed in subsequent blood samples with a concomitant increase in immature (OKT4+, OKT8+) lymphocytes that suggested an increase in release of such lymphocytes from the thymus. Assessment of 29 longterm renal allograft recipients by repeated T cell monitoring over an extended period of time confirmed the findings of other investigators that an increase in the OKT4+/OKT8+ ratio was predictive of subsequent allograft rejection episodes while subnormal OKT4+/OKT8+ ratios were indicative of possible cytomegalovirus or herpes virus infections. PMID- 6290530 TI - Antigenic relationships among some animal rotaviruses: virus neutralization in vitro and cross-protection in piglets. AB - The serotype, RNA electropherotype, and cross-protection properties of rotaviruses isolated from canine, simian, porcine, and human species were compared. The bovine strain B:USA:78:1A and the canine strain C:USA:81:2 were adapted to cell culture and cloned in this study. The other viruses, i.e., simian strain S:USA:79:2, porcine Ohio State University strain P:USA:77:1, and human strain WA, were already cell culture adapted, although they were further cloned for this work. The serum neutralization test was used to classify the viruses into serotype groups. Viruses which exhibited a difference of 20-fold or greater in neutralization titer were separated into different serotype groups. In this study, four major serotype groups were found, and these groups were represented by bovine, human, porcine, and canine-simian strains. From cross-protection studies, these serotype groups were found to be significantly different. With the exception of the porcine strain, none of the viruses used as vaccines protected gnotobiotic piglets from challenge with the virulent porcine Ohio State University strain of rotavirus. Furthermore, the canine virus protected piglets from challenge with the simian virus. The RNA electropherotype confirmed that the canine and simian strains were different in eight RNA segments and eliminated the possibility that they were the same virus. From these findings, it was concluded that only viruses belonging to the same serotype group can be expected to confer cross-protection, and thus, vaccines should be made with the serotypes to which the animal is likely to be exposed. PMID- 6290531 TI - Methylcellulose media for plaque assay of murine leukemia virus. AB - When ecotropic murine leukemia virus was assayed by a methylcellulose-XC cell procedure, plaque titers showed less test-to-test variation, more uniform dose response curves, and larger plaque sizes, as compared with results of the conventional liquid overlay-XC cell test system. This assay therefore seems to be reliable and useful for the titration of ecotropic murine leukemia virus. PMID- 6290532 TI - Comparison of Rotazyme and direct electron microscopy for detection of rotavirus in human stools. AB - A total of 115 stools were examined for Rotavirus, using direct electron microscopy (EM) and Rotazyme. The overall agreement was 88.7%. Of the negative results, there was 91.95% agreement. Rotazyme reactions of three-plus or more gave a 100% agreement with EM. The Rotazyme test is a useful diagnostic aid in laboratories not capable of performing EM. PMID- 6290533 TI - Incidence of rotavirus infection in different age groups of pediatric patients with gastroenteritis. AB - An enzyme immunoassay was used to detect rotavirus in the stools of 176 pediatric patients presenting with gastroenteritis. The highest incidence of rotavirus infection was found among patients less than 1 year of age. In contrast to previously reported studies, 23% of neonates with gastroenteritis had rotavirus in their stools. This relatively easy and rapid method was helpful in the management of pediatric patients with gastroenteritis. PMID- 6290535 TI - Acetyl glyceryl ether phosphorylcholine stimulates leukotriene B4 synthesis in human polymorphonuclear leukocytes. AB - Acetyl glyceryl ether phosphorylcholine (AGEPC) and leukotriene B4 (LTB4) induce concentration-dependent neutrophil aggregation. On a molar basis, LTB4 is approximately 10 to 100 times more potent than AGEPC. AGEPC-induced aggregation is attenuated by two inhibitors of arachidonate lipoxygenation, eicosatetraynoic acid and nordihydroguaiaretic acid, and to a lesser extent by the cyclooxygenase inhibitor, indomethacin. LTB4-induced aggregation is not readily reduced by the above inhibitors of arachidonic acid metabolism. Reverse phase high performance liquid chromatography, coupled with selective ion gas chromatography/mass spectrometry, shows that AGEPC stimulates neutrophils to synthesize sufficient LTB4 to account for the AGEPC response. In addition, the rate of LTB4 biosynthesis in response to AGEPC correlates well with the rate of AGEPC- and/or LTB4-induced neutrophils aggregation, and desensitization experiments indicate that AGEPC and LTB4 cross-desensitize. These data suggest that AGEPC-induced neutrophil aggregation may be mediated by LTB4. PMID- 6290534 TI - Membrane-bound lactoferrin alters the surface properties of polymorphonuclear leukocytes. AB - Polymorphonuclear leukocytes (PMN) aggregate and avidly attach to endothelium in response to chemotactic agents. This response may be related in part to the release of the specific granule constituent lactoferrin (LF). We found by using immunohistology and biochemical and biophysical techniques that LF binds to the membrane and alters the surface properties of the PMN. Upon exposure of PMN treated with 5 micrograms/ml cytochalasin B to 2 x 10(-7) M formyl-methionine leucine-phenylalanine for 5 min, the PMN mobilized LF to their surface as observed by immunoperoxidase staining for LF. At added LF levels ranging from 4 to 15 micrograms/10(7) PMN there was a dose-dependent reduction in PMN surface charge reaching 4 mV, when the partitioning into the membrane of a charged amphipathic nitroxide spin label was measured by electron spin resonance spectroscopy, whereas transferrin was without effect. When 125I-FeLF was added to human PMN in increasing amounts and the results corrected for the residual amount of free LF contaminating the cells, the PMN were saturated with LF at concentrations between 100 and 200 nM in the medium. Human PMN bound 1.35 x 10(6) molecules per cell and the calculated value for the association constant for these receptors was 5.2 x 10(6) M-1. Additionally, 6 micrograms/ml LF served as an opsonin for rabbit MN to promote PMN uptake by rabbit macrophages, when assessed by electron microscopy, but lysozyme did not. These studies indicate that LF can bind to the surface of the PMN and reduce its surface charge. This correlates with enhanced "stickiness" leading to a variety of cell-cell interactions. PMID- 6290536 TI - Neutralization of Epstein-Barr virus by nonimmune human serum. Role of cross reacting antibody to herpes simplex virus and complement. AB - These studies were carried out to investigate the mechanism of neutralization of purified Epstein-Barr virus (EBV) by fresh human serum from normal individuals lacking antibody to the EBV viral capsid (VCA) and nuclear antigens (EBNA). Such individuals thus lack serological evidence of immunity to EBV. Although an enzyme linked immunosorbent assay (ELISA) with highly purified immobilized EBV detected low levels of IgG antibody reactive with EBV in these normal nonimmune sera, this antibody failed to neutralize EBV in the absence of complement. Studies with depleted sera and mixtures of purified complement proteins at physiologic concentrations showed that the IgG antibody and C1, C4, C2, and C3 of the classical pathway were able to fully neutralize EBV. Mixtures of the purified components of the alternative pathway at physiologic concentrations failed to neutralize purified EBV in the presence or absence of the antibody and the alternative pathway did not potentiate classical pathway-mediated neutralization. No evidence for a requirement for C8 was obtained, precluding lysis as the mechanism of neutralization. Since C3 deposition on the viral surface accompanied classical pathway activation, viral neutralization is most likely secondary to the accumulation of complement protein on the viral surface. A coating of protein on the virus could interfere with attachment to, or penetration of potentially susceptible cells. Experiments were undertaken to determine the specificity of the IgG antibody in the sera of EBV nonimmune individuals which, together with complement, neutralized EBV. Both purified EBV and herpes simplex I (HSV-1) absorbed the EBV ELISA reactivity and EBV-neutralizing activity of nonimmune sera, whereas another member of the herpesvirus group, cytomegalovirus, was inactive in this regard. HSV-1 was quantitatively more efficient than EBV in absorbing reactivity, a finding that indicates that the antibody has a higher affinity for HSV-1 than for EBV. Further absorption studies indicated that the cross-reaction occurred in both directions as EBV also absorbed HSV-1 reactive antibodies as tested in an HSV-1 ELISA. EBV was also less efficient than HSV-1 in absorbing reactivity with HSV-1. A serum lacking detectable antibodies to both EBV and HSV-1 failed to neutralize EBV. These studies cumulatively indicate that fresh serum from EBV nonimmune individuals neutralizes EBV by the combined action of a previously undescribed cross-reacting antibody apparently elicited by HSV-1 and C1, C4, C2, and C3 of the classical complement pathway. PMID- 6290537 TI - Bile acid synthesis by long-term cultured cell line established from human hepatoblastoma. AB - Bile acids in the spent medium for the cell culture were analyzed by gas-liquid chromatography and gas-liquid chromatography-mass spectrometry to determine whether human hepatoblastoma cell line could synthesize bile acids. Cholic, chenodeoxycholic, and lithocolic acids were found in the culture medium, and a portion of chenodeoxycholic acid and all of lithocholic acid were sulfated. Since the cells had been cultured in serum-free medium, it is clear that the bile acids were newly synthesized and sulfated by the cultured cells. Chenodeoxycholic acid was the main bile acid in the medium, suggesting that the cell line might predominantly synthesize chenodeoxycholic acid. On the other hand, the cells had fetal or hepatoma characters such as marked alpha-fetoprotein production. These results suggest that fetal or hepatoma type bile acid metabolism might occur in the cell line, and that the established cell line could be an useful in vitro model for the study of bile acid metabolism in hepatoma. PMID- 6290538 TI - Specific binding sites for the triiodothyronine in the plasma membrane of rat thymocytes. Correlation with biochemical responses. AB - As a prerequisite to studies of whether the plasma membrane of the rat thymocyte contains specific, saturable binding sites for the thyroid hormone 3,5,3' triiodothyronine (T(3)), a method was developed for the isolation of a plasma membrane fraction from these cells. As judged from both electron microscopic and marker enzyme studies, the fraction was composed principally of plasma membrane vesicles, was free of nuclear contaminants, and was only slightly contaminated with other subcellular components. At 37 degrees C and pH 7.4, binding of [(125)I]T(3) by the fresh membrane preparation was rapid, reaching a maximum at 5 min and then declining with time, so that by 60 min binding was virtually nil. Decreased binding with time was due to a loss of functional binding sites, but did not reflect desensitization, since the decrease in binding activity with time was independent of the presence or absence of T(3). Scatchard analysis of saturation studies revealed the presence of two binding sites, one with an apparent dissociation constant (K(d)) of 0.95 nM and a maximum capacity of 5.3 x 10(10) sites/100 mug protein, and the other with an apparent K(d) of 25 nM and a binding capacity of 1.4 x 10(12) sites/100 mug protein. Measurement of the ability of several thyronine analogues to inhibit the binding of [(125)I]T(3) revealed the following rank order of potency: l-T(3) > l-T(4) > d-T(3) = d-T(4) > l-3,5-T(2) > rT(3) > d,l-thyronine. Binding of T(3) was inhibited by the omission of calcium from the medium or by the addition of the beta adrenergic antagonist alprenolol. As judged from studies of the lower affinity binding site, these manipulations decreased the affinity, but not the number, of binding sites for T(3). The relative potencies of thyronine analogues to inhibit the binding of [(125)I]T(3) were generally parallel to their previously reported potencies in stimulating the uptake of the sugar analogue 2-deoxy-glucose (2-DG) in intact rat thymocytes in vitro. Further, the inhibition of T(3)-binding produced by l alprenolol or by excluding calcium from the medium resembled the previously reported inhibition that these manipulations produce with respect to T(3)-induced enhancement of 2-DG uptake. These findings suggest that the binding sites for T(3) present in the plasma membrane of rat thymocytes act as functional receptors linked to the stimulation of 2-DG uptake that T(3) induces in these cells. PMID- 6290540 TI - Somatotroph hyperplasia. Successful treatment of acromegaly by removal of a pancreatic islet tumor secreting a growth hormone-releasing factor. AB - A 21-yr-old woman with Turner's syndrome presented with signs and symptoms of acromegaly. The serum growth hormone (GH) (95+/-9.4 ng/ml; mean+/-SEM) and somatomedin C (11 U/ml) levels were elevated, and an increase in GH levels after glucose instead of normal suppression, increase after thyrotropin-releasing hormone (TRH) administration instead of no change, and decrease after dopamine administration instead of stimulation were observed. The pituitary fossa volume was greater than normal (1,440 mm(3)) and the presence of a pituitary tumor was assumed. After tissue removal at transsphenoidal surgery, histological study revealed somatotroph hyperplasia rather than a discrete adenoma. Postoperatively, she remained clinically acromegalic and continued to show increased GH and somatomedin levels. A search was made for ectopic source of a growth hormone releasing factor (GRF). Computer tomographic scan revealed a 5-cm Diam tumor in the tail of the pancreas. Following removal of this tumor, serum GH fell from 70 to 3 ng/ml over 2 h, and remained low for the subsequent 5 mo. Serum somatomedin C levels fell from 7.2 to normal by 6 wk postoperatively. There were no longer paradoxical GH responses to glucose, TRH, and dopamine. Both the medium that held the tumor cells at surgery and extracts of the tumor contained a peptide with GRF activity. The GRF contained in the tumor extract coeluted on Sephadex G-50 chromatography with rat hypothalamic GH-releasing activity. Stimulation of GH from rat somatotrophs in vitro was achieved at the nanomolar range, using the tumor extract. The patient's course demonstrates the importance of careful interpretation of pituitary histology. Elevated serum GH and somatomedin C levels in a patient with an enlarged sella turcica and the characteristic responses seen in acromegaly to TRH, dopamine, and glucose do not occur exclusively in patients with discrete pituitary tumors and acromegaly. This condition can also occur with somatotroph hyperplasia and then revert to normal after removal of the GRF source. Thus, in patients with acromegaly a consideration of ectopic GRF secretion should be made, and therefore, careful pituitary histology is mandatory. Consideration for chest and abdominal computer tomographic scans before pituitary surgery, in spite of their low yield, may be justified. PMID- 6290541 TI - Morphometric study of liver cell nuclei in hepatomas using an interactive computer technique: (i) nuclear size and shape. AB - Liver cells from 20 normal livers and 20 hepatomas have been studied in histological sections using an interactive computer method which measures nuclear size and shape. The variables which gave best discrimination between malignant and benign nuclei were the standard deviations of nuclear shape measurements. Though the liver is an ideal tissue for computer study it is considered that such measurements may act as a model for analysis of nuclei of other tissues. PMID- 6290539 TI - Cyclic nucleotide-induced maturation of human promyelocytic leukemia cells. AB - Myeloid differentiation in vitro is characterized by the sequential appearance of morphological, functional, and biochemical markers of maturation. We examined the effect of agents that increased the intracellular concentration of adenosine 3'5' cyclic monophosphate on the expression of these markers by human promyelocytic leukemia cells (HL60). Cells treated with 500 muM N(6),O(2)-dibutyryl adenosine 3'5'-cyclic monophosphate expressed formyl peptide and complement receptors, reduced nitroblue tetrazolium, adhered to substrate, demonstrated chemotaxis and stimulated lysosomal enzyme release, rapidly ceased proliferation, and assumed the morphology of myelocytes and metamyelocytes. Prostaglandin E(2) (100 nM) and theophyllin (500 muM) induced similar functional changes but the cells did not mature beyond the myelocyte stage. Cholera toxin (1 or 50 nM) induced formyl peptide receptor expression and adherence, but the cells did not reduce nitroblue tetrazolium, continued to proliferate, and were unchanged morphologically. Formyl peptide receptor expression was the earliest marker of these modified programs of maturation. The receptor appeared within 2 h after treatment and increased linearly for 72 h. Receptor expression was dependent on new protein synthesis. At 48 h, Scatchard analysis demonstrated 2.4 x 10(5) receptors/ cell with a K(D) of 1.3 nM. In contrast to induction of HL60 differentiation by dimethyl sulfoxide, retinoic acid, or phorbol myristate acetate, the developmental programs initiated by agents that raised intracellular adenosine 3'5'-cyclic monophosphate shared several unique features: (a) plasma membrane maturation was dissociated from morphological maturation; (b) no latent period was evident following induction the earliest membrane marker was expressed within 2 h; (c) commitment to terminal differentiation was delayed. PMID- 6290542 TI - Wilm's tumour and renal dysplasia: an hypothesis. AB - The incidence of renal dysplasia in a series of Wilms' tumours is presented. The distribution of such lesions is discussed, together with their course of development and regression. The kidney is regarded as a particularly suitable organ for studying the relation between dysplasia and neoplasia. A schema is suggested for this association with regard to Wilms' tumour. PMID- 6290543 TI - The in vitro responses of Bacteroides fragilis to Moxalactam, Cefotaxime, Cefmetazole, Josamycin and erythromycin. PMID- 6290544 TI - A model for oral dyskinesia in rats. AB - Rats subjected to bifrontal cortical ablations developed vacuous chewing movements and jaw tremors that began 22 weeks after surgery, reached a peak rate of 4 to 8 per minute, and remained stable for an 8-week observation period. Chronic haloperidol administration to rats with bifrontal cortical ablations produced 15 to 20 movements per minute that persisted for 7 weeks after drug withdrawal. Drugs given to decrease dopaminergic, cholinergic, and gamma aminobutyric acid (GABA)-ergic neurotransmission suppressed the movements, whereas cholinergic agonists increased them. This model of oral dyskinesia in rats may be useful in developing drugs for the treatment of facial dyskinesias in humans. PMID- 6290546 TI - The distribution of cholecystokinin-like immunoreactive neurons and nerve terminals in the retrohippocampal region in the rat and guinea pig. AB - The distribution of cholecystokinin (CCK)-like (CCK-L) immunoreactive cells and nerve terminals was studied in the brains from rats and guinea pigs by using antibodies to the octapeptide cholecystokinin (CCK-8). Analysis of serial horizontal and sagittal sections through the retrohippocampal region in colcicine pretreated rats revealed a relatively large number of CCK-L immunoreactive cells in the pre- and parasubiculum, subiculum, and the medial and lateral entorhinal area (EA) at all dorsal to ventral levels of the region. In the EA, the CCK positive cells were scattered in all layers without any clear pattern. Analysis of CCK-positive cells in the retrohippocampal region showed that these cells form a morphologically heterogeneous group. The types of CCK-L immunoreactive cells ranged from small (approximately 10 micrometers) round, ovoid, or fusiform to large (approximately 30 micrometers) multipolar and pyramidal. CCK-L immunoreactive nerve fibers and preterminal processes were unevenly distributed in the retrohippocampal region. The densest innervation was found in the parasubiculum, subiculum, and the ventrolateral entorhinal area. Only a few scattered fibers were detected in the molecular layers of these structures and the outer layers of the presubiculum. Within the EA and CCK innervation indicated a heterogeneous laminar distribution that was densest in layers II and IV of the medial and lateral EA and diffuse in layers I and II. In layer II the immunoreactive nerve terminals encircled the pyramidal cell bodies, while in layers IV to VI and the most ventral part of lateral entorhinal area (LEA) and the transitional area between LEA and piriform cortex the CCK processes were distributed in a netlike fashion without clear relation to the cytoarchitectural characteristics of the area. PMID- 6290545 TI - A comparison of dendritic spine number and type on pyramidal neurons of the visual cortex of old adult rats from social or isolated environments. AB - The present study determined the effect of the housing condition experienced by old adult male rats on the appearance and number of dendritic spines. Specifically, 20-month-old rats were killed following 6 months of living in either a social environment (three to a cage) or living alone. The total number of dendritic spines per unit length was examined along segments of oblique, basal, and apical dendritic branches of pyramidal cells from layers II, III, Va, and Vb of the visual cortex. In addition to determining the total spine number, the spines were differentiated into two topographical categories: those with a lollipop configuration (type L) and those with a nubbin configuration (type N). Our results show that neither the total spine density nor the type L spine density were generally influenced by the two housing conditions. However, the density of type N spines was almost always greater on neurons from rats which had been living alone irrespective of the cortical layer or the dendritic segment counted. Some differences in total spine density and type L spine density were noted when neurons from the same environment but different cortical layers were compared, and these findings are discussed. However, the major focus of this paper was to extend our previous report of a selective increase in type N spines with age. We now show that in addition to increasing with age, type N spine density is also selectively increased by the condition of social deprivation. PMID- 6290547 TI - Histochemical evidence for sympathetic innervation of hair receptor afferents in cat skin. AB - The goal of the present study was to demonstrate histochemically fine catecholamine-containing sympathetic nerve fibers which might be associated with hair receptor afferent fibers in cat skin. Positive results would be consistent with the hypothesis that the sympathetically mediated desensitization of hair afferents, observed in this an earlier studies, is mediated by direct neurotransmitter release onto the afferent fibers. Activity in single primary afferent fibers was recorded electrophysiologically during mechanical stimulation of hairs and electrical stimulation of the lumbar sympathetic trunk in anesthetized cats. The mechanical threshold for activation was seen to increase markedly during low-frequency sympathetic stimulation. Fresh-frozen sections of skin from cat hindleg were treated with glyoxylic acid for detection of fluorescent catecholamines. Fluorescent fibers were observed in association with hair follicles as well as with arterioles and arrector-pili muscles. Subsequent silver staining of the identical skin sections revealed that the fine catecholamine-containing fibers located near hair follicles were closely associated with hair receptor afferent fibers. The results of hair receptor sensitivity is mediated by direct neurotransmitter release onto the afferent fibers. PMID- 6290548 TI - Paget's disease of the male breast. AB - Paget's disease of the breast is a malignant lesion consisting of Paget cells in the epidermis and an underlying ductal carcinoma of the breast tissue. It is estimated that perhaps 3% of female breast carcinoma cases are Paget's disease. If one estimates male breast carcinoma to be approximately 1% of the rate in the female population, then Paget's disease in men is clearly a rare clinical event. Although Paget's disease has an epidermal component, little attention has been given to this entity in the dermatologic literature. We present a case report of this disease in an elderly male patient and comment on the clinical features which should alert the physician to the presence of the malignancy. PMID- 6290549 TI - Hidroacanthoma simplex: an ultrastructural study. PMID- 6290550 TI - Functional compartments in cyclic nucleotide action. AB - Cyclic AMP-dependent protein kinase (cAMP-PK) is a ubiquitous enzyme that, when activated by cAMP, is capable of phosphorylating a variety of intracellular proteins. The central postulate of cAMP-mediated hormone action is that hormones regulate intracellular cAMP concentration and cAMP-PK mediates the effects of this second messenger. Although this postulate accurately describes cAMP action in certain systems, it does not adequately provide for recent observations of the accumulation of cAMP and the activation of protein kinase without the anticipated effects on protein kinase's substrates. Both biochemical and cytochemical technics provide evidence that hormonally-specific regulation of cAMP action occurs and is important. Our thesis is that hormonal regulation of metabolic events via cAMP is localized intracellular phenomenon. We propose that occupation of some cell-surface hormone receptors leads to cAMP accumulation and the activation of protein kinase in subcellular compartments, with the consequent phosphorylation of specific, rather than all, substrates of protein kinase. circumstances potentially contributing to this specificity include: (a) physical and kinetic compartmentation of hormone-receptor-adenylate cyclase complexes non randomly within the cell membrane; and, (b) a fixed spatial relationship of hormonally activated adenylate cyclase and specific intracellular regions by the participation of cytoskeletal proteins. PMID- 6290551 TI - Rabbit kidney cortex phosphorylase phosphatases: evidence for complexes between high molecular weight forms and heat-stable inhibitor proteins. AB - Two forms of high molecular weight phosphorylase phosphatase have been partially resolved by gel filtration chromatography of rabbit kidney cortex extracts. Two heat-stable inhibitor proteins co-eluted with the phosphatase peaks. Phosphorylase phosphatase and heat-stable inhibitor activity also co-migrated on gel electrophoresis of cortex extracts. When extracts were heated to 95 degrees for 5 minutes prior to gel filtration or electrophoresis, phosphorylase phosphatase inhibitor activity eluted at a lower molecular weight and a higher mobility, respectively. Storing cortex extracts at -20 degrees for 6 months resulted in partial conversion of both phosphatase and inhibitor activities to lower molecular weight forms which co-eluted on gel filtration. The two inhibitor peaks from gel filtration chromatography were heat-treated and characterized. Both inhibitor peaks had molecular weight of 25,000 to 35,000. The inhibitory activity of one of the peaks was increased about 3.5-fold by incubation with cyclic AMP-dependent protein kinase and ATP, and required higher concentrations of TCA to be precipitated. Hence, one of the inhibitor peaks resembled rabbit muscle inhibitor -1, while the other peak may represent an inhibitor similar to rabbit muscle inhibitor -2. These results represent the first indication that low molecular weight heat-stable inhibitor proteins may be bound to high molecular weight phosphorylase phosphatases in the cell. PMID- 6290552 TI - Melanotic neuroectodermal tumor of infancy and postsurgical dental complications: report of case. PMID- 6290553 TI - Asymptomatic, nonulcerated swelling of the posterior hard palate. AB - It is very important for clinicians to understand that nonulcerated, firm, dome shaped, nonpainful palatal swellings, if not inflammatory in nature, are probably arising in the palatal accessory salivary glands. The differential diagnosis must include benign mixed tumors; adenoid cystic carcinoma, and necrotizing sialometaplasia, which is usually ulcerated. Statistically, the possibility that the lesion is malignant is slightly greater than the possibility that it is benign. Incisional biopsy must be performed on these lesions to determine the proper treatment and management regimen. PMID- 6290554 TI - The guinea pig model of diisocyanate sensitization. I. Immunologic studies. AB - Two strains of guinea pigs were parenterally immunized with well-characterized diisocyanate-protein conjugates. Hapten-specific IgE antibodies were detected in the sera of English short-hair strain guinea pigs immunized with either toluene diisocyanate-human serum albumin (TDI-HSA) or hexamethylene diisocyanate-HSA (HDI HSA) when these sera were analyzed by the 168 hr passive cutaneous anaphylaxis (PCA) technique followed by intravenous challenges with conjugates of respective ligands coupled to an unrelated carrier protein, transferrin. IgG1 antibodies and precipitating antibodies were demonstrated in Hartley strain guinea pigs immunized with TDI/HDI-HSA conjugates. The hapten specificity of these antibodies was proved by PCA inhibition experiments and antibody absorption experiments. In the precipitating antibody system, this was further confirmed by immunoelectrophoretic analysis. Cross-reactivity between HDI and TDI was not observed in the PCA experiments. However, apparent cross-reactivity in the double gel diffusion experiments was due to new antigenic determinants formed by isocyanates after conjugation with proteins. It was therefore apparent that immune responses of guinea pigs immunized with protein conjugates of bifunctional isocyanates were heterogeneous and involved multiple specificities for hapten, carrier protein, and new antigenic determinants. It was postulated that the complex nature of the immune response generated by diisocyanate compounds in the guinea pig may also serve as a more appropriate model of isocyanate-induced human sensitivity reactions, which are known to involve diverse immunologic and nonimmunologic mechanisms. PMID- 6290556 TI - Small cell lung carcinoma. PMID- 6290555 TI - The guinea pig model of diisocyanate sensitization. II. Physiologic studies. PMID- 6290557 TI - Proceedings of the Symposium on the Physiology of the Autonomic Ganglia. Kiev, October 12-16, 1981. PMID- 6290558 TI - Voltage-dependent actions of short-chain polymethylene bis-trimethylammonium compounds on sympathetic ganglion neurons. AB - Effects of polymethylene bis-trimethylammonium compounds (with 4-7 carbons in the polymethylene chain, C4-C7) on voltage-dependence of fast excitatory postsynaptic current (EPSC) were studied in voltage-clamped neurons of the isolated rabbit superior cervical ganglion. All these compounds shortened the EPSC decay (which remained single-exponential) and decreased (or reversed) the dependence of the EPSC decay on membrane hyperpolarization. All drugs slightly decreased the EPSC amplitude; in addition, C6 and C7 decreased their dependence on membrane hyperpolarization. It is suggested that shortening of the EPSC decay produced by ganglion-blocking agents results from their binding to the open ionic channel (channel-blocking effect). The ratio of channel-blocking activities of these drugs correlates with the well-known ratio of their ganglion-blocking activities. It is suggested that the channel-blocking activities of polymethylene bis trimethylammonium compounds determine their ganglion-blocking activities. The model of channel-blocking action is discussed. PMID- 6290559 TI - Modulation of nicotinic transmission by biogenic amines in bullfrog sympathetic ganglia. AB - Studies of transmission in isolated paravertebral sympathetic ganglia of the bullfrog and at the sciatic-sartarius muscle synapse in the frog yielded evidence that biogenic amines such as catecholamines or 5-hydroxytryptamine can modulate transmission in sympathetic ganglia and at the neuromyal junction. These two transmitter substances are able to modulate transmission by affecting the amount of ACh release from presynaptic terminals and also by affecting the sensitivity of nicotinic Ach receptors of the subsynaptic membrane. Information is presented as to how these compounds exert their modulatory effects on these synapses. PMID- 6290560 TI - Intracellular recordings from lateral horn cells fo the spinal cord in vitro. AB - Intracellular recording was used in studies of the preganglionic neurons of the autonomic nervous system. These were carried out on isolated segments of the cat spinal cord. It was found that the lateral horn cells have electrical membrane characteristics similar to postganglionic neurons, but many of them have a much longer afterhyperpolarization. 5-Hydroxytryptamine, noradrenaline and aspartate induce depolarizations in lateral horn cells which are characteristically associated with an increased membrane resistance. EPSPs in lateral horn cells are not cholinergic in nature, though many cells are endowed with excitatory nicotinic receptors. Some interneurons also appear to have excitatory muscarinic receptors. Glutamate can depolarize many lateral horn cells. This excitatory amine does not seem to be responsible for the production of an EPSP, since the EPSP persisted during the continued presence of glutamate in the superfusing medium. All the neurons examined in the lateral horn are susceptible to the hyperpolarizing and shunting actions of GABA and glycine. In a small group of neurons, noradrenaline caused a hyperpolarization. PMID- 6290561 TI - Hormone regulation of adrenaline and noradrenaline release in the inferior mesenteric ganglion of the dog. AB - Studies were carried out on perfused sympathetic ganglia to determine the actions of hormones on their states and functions. The effects of hormones, hydrocortisone, cortin, prostaglandin E2 (PGE2), insulin, and triiodothyronine, on the release of adrenaline (A) and noradrenaline (NA) into the superfusate of the inferior mesenteric ganglion (IMG) of the dog were studied in rest and during 3 stimulations: (1) electrical stimulation of lumbar splanchnic and (2) hypogastric nerves; and (3) after acetylcholine application. Hydrocortisone and PGE2 do not affect A and NA release during all stimulations. Cortin increases the process during (2) and (3). Insulin increases the release of A, though not affecting NA during stimulations 1-3. Triiodothyronine increases the release of NA and decreases that of A during all 3 influences. The mechanism of hormonal action on catecholamine (CA) release by the ganglion's adrenergic structures is discussed, and the presence of insulin receptors in them is suggested. PMID- 6290562 TI - Extracellular recording of neuronal activity of the cat heart ganglia. AB - An analysis of results of extracellular recording of neuronal activity of the cat heart ganglia showed that neurons of the intracardiac ganglia, in contrast to the neurons of other visceral ganglia, were characterized by the absence of spontaneous electrical activity. Variations in the duration and frequency of mechanically induced spike discharge are indicative of a different degree of neuronal membrane depolarization and its stability in time after mechanical action of the tip of the recording microelectrode on the cell. The evoked electrical activity was not dependent on the rhythm of local contractions of the myocardium. Some evidence of interneuronal interaction at the ganglion level under the conditions of complete isolation of the right auricle has been presented. PMID- 6290563 TI - The ultrastructural organization of sympathetic ganglia of the cat. AB - The organization and relations between nerve, glial, chromaffin and other structural components of sympathetic ganglia of the cat were studied electron microscopically. Cholinergic, adrenergic and, probably, peptidergic terminals of axons making different contacts with the surrounding ganglionic cells and processes are shown with electron histochemical methods. Ultrastructural features of some dendrites are revealed which characterize them as sensory terminals of sympathetic ganglia. Besides the known axo-dendritic, axo-somatic and axo-axonal synapses, the ganglia show axo-glial synapses, and also dendro-dendritic and dendro-somatic contacts. It is established that preliminary destruction of ganglionic connections with the central nervous system is fatal only to a proportion of these interneuronal contacts while the rest of them persist. Synaptic complexes are found showing contacts of several axons and dendrites and representing a morphological substrate for convergence and divergence. No synapses are found whose presynaptic pole would be formed by a chromaffin cell. The functional role of these cells in the ganglia is discussed. PMID- 6290564 TI - Transmission of impulses from pre- to postganglionic vasoconstrictor and sudomotor neurons. AB - Transmission of impulses of pre- to postganglionic neurons supplying skeletal muscle and skin of the cat's hindlimb and tail was investigated. The objective of the study was to determine whether these postganglionic neurons can be influenced from the preganglionic side by non-nicotinic synaptic mechanisms in the lumbar sympathetic chain ganglia. The activity of the postganglionic neurons was recorded from their axons being isolated from peripheral skin and muscle nerves. (1) Vasoconstrictor neurons can be activated by muscarinic action of released acetylcholine and by a non-cholinergic synaptic mechanism. This type of non nicotinic excitation of postganglionic vasoconstrictor neurons requires the activation of thin, probably unmyelinated preganglionic axons and considerable summation. Postganglionic sudomotor and pilomotor neurons cannot be activated in this way. (2) Ongoing activity in postganglionic vasoconstrictor neurons, but not in sudomotor neurons, can be enhanced for up to 60 min by brief trains of stimuli applied to the preganglionic site. Also this enhancement requires the activation of thin preganglionic axons. (3) Stimulation of thin preganglionic axons leads to an activation of muscle vasoconstrictor neurons via non-nicotinic synaptic mechanisms in the ganglia after complete block of nicotine transmission. (4) Postganglionic vasoconstrictor neurons and sudomotor neurons may be inhibited by a catecholaminergic autogenic mechanism in the ganglia. (5) The results indicate that integration may take place in the sympathetic chain ganglia by other than divergent and convergent processes. In this integration muscarinic actions of released acetylcholine and non-cholinergic synaptic mechanisms may be involved. PMID- 6290565 TI - Intracellular localization of basement membrane precursors in the endodermal cells of the rat parietal yolk sac. I. Ultrastructure and phosphatase activity of endodermal cells. AB - The parietal layer of the rat yolk sac includes a 5 microliter thick sheet known as Reichert's membrane that exhibits properties of basement membranes. Its inner side is lined by a single layer of loosely distributed cells referred to as endodermal cells. Both Reichert's membrane and endodermal cells were examined at 13-14 days' gestation with emphasis on the ultrastructure of the Golgi apparatus, the identification of its component parts by specific phosphatase activities, and its possible role in the cells' secretory process. Reichert's membrane is composed of a series of stacked layers similar to basal laminae and composed of a network of fibrils with a diameter of 2-8 nm along which dots are located at irregular intervals. The endodermal cells contain the usual organelles, including interconnected rough endoplasmic reticulum (rER) cisternae and a prominent Golgi apparatus. With the help of phosphatase reactions, the stacks of Golgi saccules were divided into a) "phosphatase-free" saccules, the first ones on the cis or forming side, b) one or two "intermediate" saccules in the middle of the stacks, containing nicotinamide adenine dinucleotide phosphatase activity, c) one or two "last" saccules rich in thiamine pyrophosphatase activity on the trans or mature side, and d) continuing beyond the trans side, the GERL element displaying acid phosphatase activity. The latter is associated with profiles equally rich in acid phosphatase and tentatively considered to be prosecretory granules. Finally, the ectoplasm adjacent to Reichert's membrane displays large, acid phosphatase containing structures tentatively considered to be secretory granules. Thus, the extensive rER network, the well-compartmentalized Golgi apparatus, and the presence of structures which may be prosecretory and secretory granules indicate that the endodermal cells are well-equipped for the secretion of the components of Reichert's membrane. PMID- 6290566 TI - A comparison of the virucidal properties of chlorine, chlorine dioxide, bromine chloride and iodine. AB - Chlorine dioxide, bromine chloride and iodine were compared with chlorine as virucidal agents. Under optimal conditions all disinfectants were effective at low concentrations, but each disinfectant responded differently to acidity and alkalinity. Disinfection by chlorine was impaired by the presence of ammonia, but the other disinfectants retained much of their potency. Disinfection of poliovirus by iodine resulted in structural changes in the virions as seen by electron micrroscopy, but the other disinfectants were able to inactivate poliovirus without causing any apparent structural changes. PMID- 6290568 TI - The inhibitory effect of polymyxin B on endotoxin-induced endogenous pyrogen production. AB - The effect of polymyxin B (PMB) on the endogenous pyrogen (EP)-induced property of lipopolysaccharide (LPS) in vitro was examined. PMB inhibited LPS when added to leukocyte suspension 5 min before or up to 30 min after the addition of LPS. The inhibitory effect was dose-related and appeared to be specific for LPS (including naturally occurring endotoxin). EP production in response to a different stimulus (staphylococci) was not prevented even when LPS-PMB complexes were presumably present. These data suggest that when experimental agents are found to stimulate the production of EP or lymphocyte activating factors (LAF, interleukin-1) in vitro, or when apparently spontaneous production of EP or LAF is seen, incubation with PMB may be a useful technique to exclude th effects of endotoxin contamination - especially when negative results have been obtained in the limulus gelation test. PMID- 6290567 TI - A non-lethal test of the resistance of inbred mice to herpes simplex virus type 1. AB - Inbred strains of mice varying in susceptibility to intraperitoneal (i.p.) inoculation of HSV-1 were tested by inoculating 10(5) p.f.u. of the SC 16 strain into the ear pinna. Increase in ear thickness was less in resistant than in susceptible strains. In the resistant C57BL/6 strain, local replication of virus, spread to cervical ganglia and development of latent infections of the ganglia were all less than in susceptible DBA/2 mice; there was also less cellular infiltration of the inoculation site. Like resistance to i.p. inoculation, resistance to ear inoculation appears to be inherited as a dominant characteristic. The test provides a non-lethal method of distinguishing between resistance and susceptibility of inbred strains of mice to HSV-1 and may also be useful in defining resistance factors. PMID- 6290570 TI - Hyporesponsiveness to the immunosuppressant effects of delta-8 tetrahydrocannabinol. AB - Delta-9-tetrahydrocannabinol (delta 9-THC) and delta 8-THC have previously been shown to suppress humoral immunity in mice. The purpose of these investigations was to determine whether a hyporesponsiveness develops to the immunosuppressive effect of delta 8-THC. BALB/c mice were administered 60 mg/kg delta 8-THC i.p. 2 days after an i.p. immunizing dose of sheep erythrocytes (sRBC). Significant inhibition of direct hemolytic plaque-forming cells (PFC)/spleen was observed on days 3--6, with peak day of inhibition occurring on day 4. The effective inhibiting dose 50 (ED50) measured on peak day of response was 40 mg/kg for PFC/10(6) spleen cells and 38 mg/kg for PFC/spleen. When mice were pretreated daily for 5 days with different doses of delta 8-THC, similar ED50's were detected. Under this pretreatment regimen, 5 or 10 mg/kg produced no immunosuppression. Daily treatment with 5 or 10 mg/kg delta 8-THC for 5 days prior to sRBC and varying doses of delta 8-THC administered 2 days after sRBC resulted in significantly higher ED50's and increased values for the slopes of the dose response curves. These results suggest that a hyporesponsiveness develops to the immunosuppressive activity of delta 8-THC. PMID- 6290569 TI - The metabolism of adenosine and distribution of adenosine receptor lymphocytes in two human circulating T cell subsets. AB - Human circulating E rosette forming cells (ERFC), rerosetted with sheep erythrocytes in the presence of adenosine, yielded two T-lymphocyte subpopulations: a major fraction forming E-rosettes (E resistant = ER) and a minor non-rosetting fraction (E sensitive - ES). Both T cell subpopulations converted adenosine mainly into inosine. However, ES cells metabolized adenosine more extensively than ER cells. Adenosine deaminase (ADA) activity was significantly higher in ES cells. Purine nucleoside phosphorylase (PNP) activity, as well as hypoxanthine guanosine phosphoribosyl transferase (HGPRT) activity were similar in both T cell subsets. The ratio of ADA/PNP in ES cells relative to ER cells was 1.8 suggesting that ES cells are at an earlier stage of differentiation. Enrichment of lymphocytes bearing a receptor for adenosine was demonstrated in ES cells. PMID- 6290571 TI - Lipolymph node. PMID- 6290572 TI - The receptor concept: basic to clinical science. PMID- 6290573 TI - Herpetic proctitis and meningitis: recovery of two strains of herpes simplex virus type 1 from cerebrospinal fluid. AB - A patient with simultaneous proctitis and meningitis due to herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2) was extensively investigated. In both disease locations the infection was clinically evident and culture-proven. Analysis by sodium dodecylsulfate-polyacrylamide gel electrophoresis of rectal isolates revealed both HSV-1 and HSV-2. The cerebrospinal fluid harbored two apparently different strains of HSV-1, one of which was shown by restriction endonuclease analysis to be identical with the rectal isolate of HSV-1. PMID- 6290574 TI - Entamoeba histolytica cytotoxin: purification, characterization, strain virulence, and protease activity. AB - A heat-labile cytotoxin was isolated from virulent strains of axenically cultivated Entamoeba histolytica. Strains of E. histolytica representing a spectrum of virulence as determined in animal and in vitro models of disease were examined for cytotoxic activity. Extracts of virulent strain HM1 possessed marked cytotoxic activity, those of moderately virulent strain 200 showed intermediate activity, and those of avirulent strains 303 and Rahman showed no activity. The cytotoxin was partially purified from the cell-free supernatant of sonicated E. histolytica HM1 trophozoites by ammonium sulfate precipitation and gel filtration. Cytotoxic activity was stable in a narrow pH range (6-7.2) and in 1 M NaCl, urea, and guanidine. Specific immune rabbit and human antiserum as well as the protease inhibitors aprotinin, pepstatin, and leupeptin inhibited cytotoxicity. The partially purified cytotoxin did not have any detectable degradative enzymatic activities. Thus, virulent strains of E. histolytica possess an immunogenic cytotoxic protein which may be important in the pathophysiology of amoebiasis. PMID- 6290576 TI - Prolonged excretion of group A coxsackievirus in an infant with agammaglobulinemia. PMID- 6290575 TI - Pathogenicity of acyclovir-resistant herpes simplex virus type 1 from an immunodeficient child. AB - Sequential isolates of herpes simplex virus type 1 (HSV-1) from a child with severe combined immunodeficiency were examined for sensitivity to acyclovir. Early intravenous courses of acyclovir resulted in dramatic clinical improvement and were associated with the isolation of sensitive strains of HSV-1 (ID50[dose inhibiting 50% of control plaques], 0.010-0.106 microgram/ml), whereas later recurrences following intravenous, oral, and ophthalmic therapy were characterized by low-grade chronic lesions (ID50, 1.04-9.43 microgram/ml) that were unresponsive to acyclovir despite serum levels of up to 10.45 microgram/ml. Diminished sensitivity was associated with reduced viral thymidine kinase activity, and linked resistance with idoxuridine was detected in the isolates from the patient's eye. Intracerebral and cutaneous snout inoculation of a resistant isolate into BALB/c, hairless, and athymic nude mice revealed a 100- to 1,000-fold decrease in virulence as compared with an early sensitive isolate. Acyclovir-resistant HSV-1 can emerge in certain clinical settings but may be associated with diminished virulence. PMID- 6290577 TI - Alteration of diabetes and interferon induction by the diabetogenic strain of encephalomyocarditis virus through cell passage. AB - The diabetogenic strain of encephalomyocarditis virus (D virus) was propagated in several continuous cell lines. Each virus stock was tested for its ability to produce diabetes in mice and induce L-cell interferon (IFN-beta). The effect of insulin on virus replication and IFN-beta induction was also determined. It was found that the severity of the diabetes and the amount of IFN-beta produced was dependent on the cell line used for virus propagation. Virus synthesis was augmented and IFN-beta production was altered in insulin-treated cell cultures. It is concluded that D virus either consists of more than one virus or that its diabetes and IFN-beta inducing characteristics are unstable. PMID- 6290578 TI - Mechanism of interferon action: eIF-2 alpha phosphatase in interferon-treated mouse fibroblasts is double-stranded RNA independent. AB - The effect of double-stranded RNA on the dephosphorylation of purified, 32P labeled eIF-2 was examined in cell-free extracts prepared from interferon-treated mouse L929 cells. Dephosphorylation of the alpha subunit of eIF-2 occurred at comparable rates in the presence and in the absence of reovirus dsRNA. By contrast, the beta subunit of eIF-2 was not dephosphorylated to any significant extent in either the presence or the absence of dsRNA. These results indicate that the enhanced phosphorylation of eIF-2 alpha observed in IFN-treated systems in the presence of double-stranded RNA (dsRNA) is indeed caused by an activation of a protein kinase rather than to an inhibition of a phosphoprotein phosphatase by the dsRNA. PMID- 6290579 TI - [A case of hepatoma associated with Vibrio vulnificus bacteremia and its bacteriological findings]. PMID- 6290580 TI - [Seroepizootiological studies on type A influenza virus against porcine serum in Tochigi Prefecture over a period from July, 1979 to June, 1980]. PMID- 6290581 TI - [Biosynthesis of vitamin D]. PMID- 6290582 TI - [Studies on the mechanism of human parturition]. PMID- 6290583 TI - [Liver diseases. C 1. Treatment of liver cancer]. PMID- 6290584 TI - [Extra-nasopharyngeal extensions of angiofibroma]. AB - Four cases of NPA with extra-nasopharyngeal extensions have been presented, two of which had an intracranial extension. On of the cases with intracranial extension underwent a transtemporal craniotomy removal and four temporal fossa. The other three cases were managed by the sublabial and transantral routes. Methods of pre-operative diagnosis of such extensions have been discussed and the importance of pre-operative recognition of extension in every NPA has been emphasized. Various approaches to the lateral and intracranial extensions have been reviewed. We feel that, for lateral extensions, sublabial and/or transantral or transzygomatic approaches are adequate, while the radical approach of Karnik is to be preferred only in large lateral extensions and in cases of intracranial extensions without symptoms of a space-occupying intracranial lesion. However, for patients with intracranial symptoms, a staged procedure is indicated and is safe for the patient. PMID- 6290585 TI - Comparison of the effects of prostaglandin E2, prostacyclin and 1-24 adrenocorticotrophin on plasma cortisol levels of fetal sheep. AB - The changes in plasma cortisol levels in response to intravenous infusions of prostaglandin E2 (PGE2), prostacyclin and 1-24 ACTH have been studied in chronically catheterized fetal sheep during the last third of gestation. All three drugs increased plasma cortisol levels with prostacyclin being sigificantly more potent than either PGE2 or 1-24 ACTH. No interaction between the steroidogenic actions of 1-24 ACTH and either PGE2 or prostacyclin could be demonstrated. The steroidogenic action of PGE2 was not significantly modified by fetal hypophysectomy. It is concluded that neither PGE2 nor prostacyclin is likely to be involved in the enhanced adrenal responsiveness to 1-24 ACTH observed in fetal sheep in the period immediately before birth. PMID- 6290586 TI - Effects of 2-hydroxyoestradiol-17 beta on plasma luteinizing hormone, follicle stimulating hormone and prolactin, and nuclear translocation of pituitary oestrogen receptors in ovariectomized ewes. AB - The ability of oestradiol-17 beta (OE2) and 2-hydroxyoestradiol (2OH-OE2) to translocate pituitary oestrogen receptors to the nuclear compartment and to affect plasma concentrations of LH, FSH and prolactin was studied in ovariectomized ewes. Mean (+/- S.E.M.) nuclear oestrogen receptor levels (% of total pituitary oestrogen receptors) after intracarotid injections of 1.25 micrograms OE2, 400 micrograms 2OH-OE2 or vehicle were 8.7 +/- 2.3, 16.9 +/- 3.2 and 0.8 +/- 0.1% respectively. Whereas 1.25 or 12.5 micrograms OE2 significantly lowered plasma LH and FSH, 400 micrograms 2OH-OE2 did not affect plasma LH or FSH levels. Injection of 4000 micrograms 2OH-OE2 however, significantly affected plasma LH and FSH. Plasma prolactin levels were not significantly affected by the treatments. These data indicate a discrepancy between oestrogen receptor occupancy and effects on gonadotrophin and prolactin secretion after injection of catechol oestrogen. PMID- 6290587 TI - A rapid direct radioimmunoassay for the measurement of oestrone sulphate in the milk of dairy cows and its use in pregnancy diagnosis. AB - A radioimmunoassay for oestrone sulphate in unextracted samples of milk has been developed. The assay was validated by comparison with a method involving hydrolysis and extraction. The direct assay was used to measure oestrone sulphate in milk samples taken at weekly intervals throughout pregnancy in a commercial dairy herd. Concentrations of oestrone sulphate increased approximately 100 days after insemination and were maintained throughout the remainder of pregnancy in the range of 1.85-3.70 nmol/l. PMID- 6290588 TI - The acid-base responses of gills and kidneys to infused acid and base loads in the channel catfish, Ictalurus punctatus. AB - Acid and base loads infused into the dorsal aorta of the freshwater channel catfish (Ictalurus punctatus Rafinesque) were excreted by both gills and kidneys. The gills excreted 2-3 times as much as the kidneys. After a 2 mmol . kg-1 load of NaHCO3, the gills excreted 50% of the net OH- load in the first 5 h, while the kidneys excreted 18% in the first 6 h. After a 2 mmol . kg-1 load of NH4Cl, the gills excreted 32% of the load in 6 h, and 16% was excreted renally in 20 h. There was no evidence of tissue damage after either NaHCO3 or NH4Cl infusions, whereas infusion of 2 mmol . kg-1 HCl or 1 mmol . kg-1 of L(+)-lactic acid caused significant kidney damage and extensive tissue necrosis within 24 h. After both NaHCO3 and NH4Cl infusions, the majority of the load had been transferred to the intracellular compartment within 2 h. From there it was excreted slowly, presumably by transfer back through the extracellular compartment. Due to the relative compartment volumes and buffer values, the change in intracellular pH was less than 0.05 units, while the blood pH was changed by as much as 0.3 units. PMID- 6290589 TI - Structure and expression of endogenous ecotropic murine leukemia viruses in RF/J mice. AB - High leukemic mouse strains possess proviral genomes that are more inducible for virus expression by halogenated pyrimidines than the proviral genomes harbored by low leukemic mice. We investigated the induction and arrangement of ecotropic proviruses in RF mice, a strain of mouse that develops a moderate incidence of leukemia late in life. We found that RF mice, unlike either high or low leukemic inbred strains, carried both a gene for high efficiency virus induction (Rjv-1) and a gene for low efficiency virus induction (Rjv-2). Virus induction from mice that contained Rjf-2 alone was observed only in crosses with two other strains that carried ecotropic proviruses, i.e., DBA/2 and C57BL/6, and not in crosses performed with mice that lacked ecotropic proviruses, i.e., 129, SWR, and NFS. Inheritance of the Rjv-1 gene frequently resulted in viremia when a virus suppressive gene(s) of RF (most likely Fv-1) was not present in the same individual. Rjv-1 and Rjv-2 virus induction genes co-segregated with ecotropic proviruses integrated in different cellular DNA sequences. Rjv-2, the less inducible ecotropic provirus in RF mice, is located in cellular DNA sequences very similar to those found adjacent to the ecotropic provirus of BALB/c. These results document a second system of virus interaction or complementation and demonstrate that ecotropic proviruses of different phenotypes can be found within an individual mouse strain. PMID- 6290590 TI - Increased urinary excretion of angiotensin converting enzyme in patients with renal diseases. AB - We measured the activity of angiotensin converting enzyme (ED 3.4.15.1) in urine samples from normal subjects and patients with nephrotic syndrome and chronic glomerulonephritis. Urinary excretion of angiotensin converting enzyme in patients with nephrotic syndrome (1.58 +/- 0.50 (SD) units/day; n = 15) and chronic glomerulonephritis (1.01 +/- 0.45 units/day; n = 12) was significantly increased compared with normal subjects (0.38 +/- 0.10 units/day; n = 18). It was demonstrated that a high molecular weight form of the enzyme (more than 400000) was mainly excreted in urines from patients with nephrotic syndrome. After trypsin treatment, this form was altered to a low molecular weight form (290000). These two different forms of urinary angiotensin converting enzyme were compared with the enzyme purified from human kidney, and found to be identical with respect to Km values (2 mmol/l), pH optimum (pH 8.3), chloride ion dependency (0.8 mol/l), inhibitory effect of captopril (I50, 21 nmol/l), and behavior towards antiserum to human kidney angiotensin converting enzyme. PMID- 6290591 TI - DNA sequence analysis of an immediate-early gene region of the herpes simplex virus type 1 genome (map coordinates 0.950 to 0.978). AB - The nucleotide sequence of 4 kilobases of DNA from within the short region of the genome of herpes simplex virus type 1 has been determined. This portion of DNA contains the junctions of the terminal and inverted repeat sequence components with the unique sequence component and the 5'-regions of the genes which encode the Vmw 12, Vmw 68 and Vmw 175 immediate-early polypeptides. The transcription and translation initiation sites of all three genes and the 5' and 3' boundaries of the Vmw 12 and Vmw 68 gene introns have been localized on the DNA sequence and shown to be flanked by sequences which resemble those found in similar positions in other eukaryotic genes. For the Vmw 12 and Vmw 68 genes the promoters, the 5' non-coding regions of the mRNAs, and the introns lie within the terminal and internal inverted repeats respectively while the polypeptide-coding regions lie in the short unique component. The introns consist largely of tandemly reiterated copies of a 22-nucleotide sequence: the Vmw 12 gene intron contains seven of these and the Vmw 68 gene intron five. The Vmw 175 gene is located entirely within the short repeats, of which those regions sequenced here have the extremely high G + C content of 78%, in marked contrast to the value of 66% obtained for the adjacent region of the unique sequence component. Prediction of the complete amino acid sequence of the Vmw 12 polypeptide, accounting for a mol. wt. of 9830, and of the first 523 amino-terminal amino acids of the Vmw 175 polypeptide has been made from the DNA sequence. The polypeptide-coding region of the Vmw 175 gene has an average G + C content of 80% but nevertheless specifies a wide range of amino acid types because of the maximal assignment of G and C residues to colon third base positions. PMID- 6290592 TI - Further observations on the ultrastructure of human rotavirus. AB - The inner capsid layer of human rotavirus was found to preferentially break up into large ring-like morphological units. The outer capsid was found to be composed of capsomeres covered by a thin protein or glycoprotein covering. These capsomeres appeared to be broad headed and short stemmed, similar to the type of pin used to mark locations on a map (pushpin). PMID- 6290593 TI - Genomic location and lack of phosphorylation of the HSV immediate-early polypeptide IE 12. AB - Herpes simplex virus type 1 (HSV-1) induces an immediate-early (IE) polypeptide IE 12. An equivalent polypeptide coded by HSV-2 which migrated slightly more slowly on SDS-polyacrylamide gels was identified and designated IE 12.3. Analysis of the serotype of the IE polypeptide induced by five HSV-1/HSV-2 intertypic recombinants when correlated with their genome structures showed that IE 12/IE 12.3 mapped in the region spanning the TRs/Us junction. Unlike four other IE polypeptides induced by HSV-1 (IEs 175, 110, 68 and 63), IE 12 was not detectably phosphorylated. PMID- 6290594 TI - Differential incorporation of thymidylate analogues into DNA by DNA polymerase alpha and by DNA polymerases specified by two herpes simplex viruses. AB - Several triphosphates (TP) of 5-substituted deoxyuridine (dU), like 5-ethyl (Et), 5-n-propyl (n-Pr), 5-iso-propyl (iso-Pr), 5-n-hexyl (n-Hx), and 5 trifluorothymidine (F3-dT) were used as substrates for HeLa DNA polymerase alpha and for two herpes simplex virus (HSV)-coded DNA polymerases isolated from HeLa cells infected with HSV-1, strain C42 (wild-type), or its mutant resistant to phosphonoformate (PFAr). All polymerases were purified up to the DNA-cellulose column step and they showed comparable specific activities. The incorporation into DNA studied with all the alkyl analogues of dUTP is several times higher with the virus enzymes than with DNA polymerase alpha. The DNA polymerase of the mutant virus incorporates dUTP analogues to a lower extent than the wild-type polymerase. The two virus enzymes also differ in the Km and Vmax values for different substrates, indicating that the mutation to PFAr has affected the structure of the virus DNA polymerase. Surprisingly, all three enzymes use F3 dTTP as substrate for DNA synthesis to an equal but limited extent. PMID- 6290595 TI - Variation in murine cytomegalovirus replication in fibroblasts from different mouse strains in vitro: correlation with in vivo resistance. AB - The in vitro growth of murine cytomegalovirus (MCMV) in mouse embryo fibroblasts (MEF) from Balb/c, C57BL, C3H and CBA mouse strains has been shown to correlate with the genetically determined in vivo susceptibility of these strains to MCMV infection. MEF from Balb/c and C57BL mice were more susceptible to MCMV than those from C3H and CBA mice. This was evident regardless of whether replication was measured by cytopathic effect (c.p.e.) score, virus yield, plaque count, plaque size or time of onset of c.p.e. The growth of MCMV in tracheal organ cultures from different mouse strains was similar to that observed in MEF from these strains. The replication in MEF when measured by c.p.e. score and virus yield was affected by the density of the cell cultures. The strain of mouse used to produce MCMV also affected the comparative sensitivity of MEF to the virus. This appeared to be due to reduced growth of MCMV in its homologous MEF type, an unexpected result. In contrast, cell density had little effect on the replication of herpes simplex virus type 1 (HSV-1), but again the in vitro susceptibility of MEF reflected the in vivo susceptibility of mouse strains to infection with this virus. PMID- 6290596 TI - A comparison of the intracellular precursor polyproteins of simian sarcoma associated virus [SiSV(SiAV)] and three human virus isolates: HL23V, HEL12V and A1476V. AB - The intracellular precursor polyproteins of simian sarcoma-simian-associated virus [SiSV(SiAV)] were compared to the intracellular proteins of the human retrovirus isolates. HL23V, HEL12V and A1476V, by radioimmunoprecipitation followed by SDS-polyacrylamide gel electrophoresis and tryptic peptide analysis. Cells infected with SiSV(SiAV) were characterized by polyproteins Pr200gag-pol, gPr80env, Pr80gag, Pr60gag and Pr40gag. Identical intracellular precursor polyprotein profiles were obtained from cells infected with HL23V, HEL12V and A1476V. Tryptic digest mapping of peptides containing [3H]leucine showed the structural composition of Pr60gag to be the virus core proteins, p28, p15/p12 and p10. The SiSV(SiAV) envelope precursor, gPr80env, contained the structural determinants of mature viral gp70 and a non-glycosylated protein termed p15E. The homology of the human isolate viruses, HL23V, HEL12V and A1476V, to the SiSV(SiAV)/GaLV (gibbon ape leukaemia virus) family of viruses was confirmed by mapping studies. Both gPr80env and Pr60gag of SiAV were identical by tryptic peptide mapping to the respective proteins from the three human retrovirus isolates examined. The potential significance of these results to considerations of the origins of SiAV and the SiAV-like human isolates is discussed. PMID- 6290597 TI - Inhibition of vesicular stomatitis virus glycoprotein expression by chloroquine. AB - Addition of 50 micrograms/ml chloroquine to neuroblastoma cells 1 h before infection with temperature-sensitive mutant ts G31 (III) of vesicular stomatitis virus (VSV) prevented virus-induced cell fusion from occurring. Interestingly, addition of chloroquine after infection still inhibited cell fusion. Based on the number of fusion events required to produce the polykaryocytes observed, cell fusion was inhibited 92% when chloroquine was added 1 h post-infection and 77% when chloroquine was added 2 h post-infection. The inhibition of virus-induced cell fusion could not be accounted for by an inhibition of virus protein synthesis because the virus protein synthesis measured 6 h post-infection was 90% of that in untreated, infected cells with chloroquine added 1 h post-infection, and the same as untreated, infected cells when chloroquine was added 2 h post infection. No virus proteins were made, however, when chloroquine was added before infection, which is consistent with a chloroquine-mediated inhibition of virus uncoating. The release of infectious virions was completely inhibited when chloroquine was added before infection or 1 or 2 h post-infection, which indicated an inhibition of virus maturation in the later stages of virus assembly. By indirect immunofluorescence the virus glycoprotein (G protein) could not be detected on the surface of chloroquine-treated, infected cells, but the G protein was present inside the treated cells. With 125I-labelled anti-G protein IgG, 16% of the G protein found on the surface of untreated, infected cells was on the cell surface when chloroquine was added 2 h post-infection. When chloroquine was removed from infected cells, the G protein accumulated at the cell surface, and this accumulation could not be prevented by tunicamycin, an inhibitor of glycosylation. Furthermore, galactose was incorporated into the G protein in the presence of chloroquine. Therefore, the VSV G protein was being synthesized and glycosylated in the presence of chloroquine but the drug prevented the expression of the glycoprotein at the cell surface during the final stages of G protein assembly.U PMID- 6290598 TI - A simple and rapid test for the identification of clinical herpes simplex virus isolates. AB - A simple procedure is described which permits the rapid identification of clinical herpes simplex virus isolates. The test utilizes Staphylococcus aureus to which anti-viral immunoglobulins had been adsorbed. Adherence of the antibody coated bacteria to virus-infected cells is readily seen by light microscopy. Indirect immuno-fluorescence and the S aureus adherence reaction were found to be approximately of equal sensitivity for the detection of virus antigens. PMID- 6290599 TI - Detection of antibodies to cytomegalovirus by an improved counterimmunoelectrophoretic procedure. AB - An improved counterimmunoelectrophoretic (CIE) procedure for detection of antibodies to human cytomegalovirus (CMV) is described. The procedure, which uses high ionic strength buffer and concentrated antigen, is as sensitive and specific as complement fixation (CF), indirect hemagglutination (IHA), and anti-complement immunofluorescence (ACIF) but requires concentration of some sera. In studies on sera from 118 normal blood donors, the percent correlation of CIE results with those by CF, IHA, and ACIF were 97.5, 98.3, and 98.3, respectively, and exceeded slightly the correlations among the latter three procedures. The appearance of precipitation immediately or early after a CIE run and of more than one line of precipitate was indicative of high titers by the other procedures, while single lines of late precipitation occurred with most low titer sera but also with many of those of high titer. PMID- 6290600 TI - Transmission of hepatitis A virus among recently captured Panamanian owl monkeys. AB - The presence of antibody to hepatitis A virus (anti-HAV) in 60% of procured owl monkeys (Aotus trivirgatus) held within the United States prompted a study of recently captured A trivirgatus in Panama. Only 2 of 145 newly captured monkeys, but all of 35 A trivirgatus held within a colony for over 100 days, were found to have anti-HAV. Of 41 sero-negative, newly captured monkeys followed prospectively, 25 became infected with hepatitis A virus (HAV) as evidenced by seroconversion or demonstration of virus in the liver at death. Only one monkey that survived over 60 days within the colony was not infected. HAV was identified in the feces of most infected monkeys prior to the development of antibody and was antigenically indistinguishable from human HAV in cross-blocking radioimmunoassays. This colony-centered epizootic provides strong evidence that A trivirgatus is susceptible to HAV and should be investigated further as a potential model of human hepatitis A. PMID- 6290601 TI - Neonatal rotavirus infection: role of lacteal neutralising alpha1-anti-trypsin and nonimmunoglobulin antiviral activity in protection. AB - Expressed breast milks (EBMs) were collected from mothers of rotavirus (RV) excreting babies and from mothers whose babies were RV free during an outbreak of asymptomatic RV infection in a newborn nursery to determine the role of lacteal anti-RV neutralising activity, alpha1-anti-trypsin activity, and nonimmunoglobulin antiviral factor in protection of neonates from RV infection, and although all of the above factors were present in the majority of the EBMs, no correlation could be found between their presence in EBM and protection from RV infection. A significant rise in both neutralising activity and subgroup 2 antibodies, was demonstrated in the EBM of one mother who experienced a subgroup 2 RV-associated diarrhoea during lactation. However, the alpha1-anti-trypsin activity and the nonimmunoglobulin antiviral levels remained the same. The importance of these factors in passive immunity with reference to virus dose is discussed. PMID- 6290602 TI - Morning injections of large doses of melatonin, but not of 5-methoxytryptamine, prevent in the hamster the antigonadotropic effect of 5-methoxytryptamine administered late in the afternoon. PMID- 6290603 TI - The ionic dependence of black widow spider venom action at the stretch receptor neuron and neuromuscular junction of crustaceans. AB - The effects of black widow spider venom (BWSV) on the crayfish stretch receptor and the lobster neuromuscular junction were examined. In crayfish stretch receptor neurons, BWSV caused a slight hyperpolarization followed by a large depolarization. The venom-induced depolarization of the stretch receptor was caused by an increase in membrane conductance to Na+ and Ca2+. Black widow spider venom also caused an increase in the frequency of miniature inhibitory postsynaptic potentials recorded in the stretch receptor. The ability of BWSV to increase the frequency of miniature excitatory postsynaptic potentials (MEPSPs) at the lobster neuromuscular junction was dependent on the divalent cation composition of the bathing medium. Ringer solutions containing Ca2+ supported the greatest venom-induced increase in MEPSP frequency, Mg2+ and Mn2+ supported a moderate increase in MEPSP frequency, while Co2+ and Zn2+ blocked this venom effect entirely. Black widow spider venom did not block axonal conduction in lobster walking leg axons or in the axon of the crayfish stretch receptor. The results suggest that in crustaceans, BWSV interacts specifically with membrane of the soma-dendritic region of the stretch receptor and with nerve terminal membrane, causing an increase in Na+ and Ca2+ conductance. PMID- 6290604 TI - All-or-none control of the bursting properties of the pacemaker neurons of the lobster pyloric pattern generator. AB - In Crustacea the central pattern generator for the pyloric motor rhythm (filtration to the midgut) is known to be located within the stomatogastric ganglion (STG); its cycling activity is known to be organized by three endogenous burster neurons acting as pacemakers and driving 11 follower neurons. In Homarus, recordings from the isolated stomatogastric nervous system (Fig. 1) indicate that (1) the pyloric output can be generated only when the STG is afferented (i.e., connected to the more rostral oesophageal and commissural ganglia) (Fig. 2) and (2) the deafferentation of the STG results in a complete loss of the bursting properties of the pacemaker neurons (Fig. 4). Manipulation of the STG inputs responsible for unmasking the properties of the pacemakers strongly suggests that (1) they are not phasic inputs (Fig. 5) and (2) they are long-term acting inputs (Fig. 6). These results provide evidence for a neural all-or-none control of the bursting properties of the pacemaker neurons of a motor pattern generator. PMID- 6290605 TI - The relationship between the size of a muscle afferent volley and the cerebral potential it produces. AB - This study examined the relationship between the size of an afferent neural input produced by electrical stimulation of the posterior tibial nerve at the ankle and the size of the early components of the evoked cerebral potential. For five of six subjects the first peak of the afferent neural volley recorded in the popliteal fossa was uncontaminated by either motor efferents or cutaneous afferents. This was established by measuring the conduction times of motor fibres in the posterior tibial nerve and cutaneous fibres in the sural and posterior tibial nerves over the ankle to popliteal fossa segment. It is likely therefore that the first peak of the afferent volley contained predominantly, if not exclusively, activity in rapidly conducting afferents from the small muscles of the foot. The size of the two earliest components of the cerebral potential did not increase in direct proportion to the size of the afferent volley which produced it. The early components of the cerebral potential reached a maximum when the responsible muscle afferent volley was less than 50% of its maximum. PMID- 6290606 TI - Early and late components in the human anal reflex. AB - Perianal electrical stimulation induces reflex activity in the superficial part of the external anal sphincter muscle. Several components of differing latency can be recognised. It is suggested that these correspond with the ripples of contraction observed clinically after perianal scratch stimuli. Some of our earlier studies suggested an erroneously short latency for the first component of this cutaneously-elicited reflex. The reasons for this are discussed. PMID- 6290607 TI - Anal sphincter responses after perianal electrical stimulation. AB - By perianal electrical stimulation and EMG recording from the external anal sphincter three responses were found with latencies of 2-8, 13-18 and 30-60 ms, respectively. The two first responses were recorded in most cases. They were characterised by constant latency and uniform pattern, were not fatigued by repeated stimulation, were most dependent on placement of stimulating and recording electrodes, and always had a higher threshold than the third response. The third response was constantly present in normal subjects. It had the longest EMG response and the latency decreased with increasing stimulation to a minimum of 30-60 ms. This response represented the clinical observable spinal reflex, "the classical anal reflex". The latencies of the two first responses were so short that they probably do not represent spinal reflexes. This was further supported by the effect of epidural anaesthesia which left the first responses unaffected but abolished the classical anal reflex. The origin of the two first responses is discussed and models involving antidromal impulse propagation in the efferent fibre as the afferent limbs of the responses are proposed. PMID- 6290608 TI - Investigations on the nervous mechanisms underlying the somatosensory cervical response in man. AB - The main features (amplitude, latency and shape) of the cervical activity evoked by stimulation of the median nerve, recorded throughout the cervical spine, have been concurrently investigated by monopolar, bipolar longitudinal and bipolar transverse recordings. In some subjects the derivation C7-Sn (suprasternal notch) has been employed as well. A comparative evaluation of the refractory period of each component of the cervical responses under investigation has been performed to differentiate presynaptic from postsynaptic events. Additional information has been obtained by cervical activity recorded by longitudinal and transverse bipolar derivations upon stimulation of the lower limb. It was thus demonstrated that both presynaptic and postsynaptic events were responsible for the cervical sensory evoked potential, as appearing when recorded against a cranial reference (that is the upper midfrontal region). The structures involved were the brachial plexus (N9), the cervical roots (P10 and a minor part of N11a), the dorsal columns both at caudal (N11a) and rostral (N11b) cervical levels, and the dorsal column nuclei (N13). However a contribution of the spinal segmental activity to the postsynaptic portion of the cervical response, more specifically to N13, should be considered as well, though direct evidence is still inadequate. PMID- 6290609 TI - Friedreich's ataxia. Early detection and progression of peripheral nerve abnormalities. PMID- 6290610 TI - Plasma cyclic nucleotide responses to methacholine in patients with Adie's syndrome. PMID- 6290611 TI - SV-40 virus-induced neoplastic transformation of hamster brain cells in vitro. Studies with scanning and transmission electron microscopy. AB - Neoplastic transformation of one-day-old hamster brain cells was produced by infection with SV-40 virus and verified by phase contrast microscopy, growth in semisolid media, and intracranial tumor production after inoculation of the cells into other one-day-old hamsters. Transformed cells were studied by transmission and scanning electron microscopy. The numerous alterations in cell surface structure and in nuclear and cytoplasmic organization suggest a marked increase in cell metabolism and in the rate of mitosis and cell division. Cilia with a nine-to-zero pattern of microtubule doublets were present in cells with intermediate size filaments which stained for glial fibrillary acidic protein. The findings indicate that infection of one-day-old hamster brain cells in culture by SV-40 virus results in their transformation to a neoplastic state and the transformed cells are differentiating neoplastic astrocytes. PMID- 6290612 TI - Hydrocephalus in weanling mice induced by a temperature-sensitive mutant of vesicular stomatitis virus. AB - Hydrocephalus developed in weanling Swiss-Webster mice after intracerebral (IC) inoculation of a naturally selected temperature-sensitive (ts) mutant of vesicular stomatitis virus (VSV). This spontaneous ts mutant was isolated from a persistent infection (pi) of mouse L cells with VSV, and named VSV-tspi 364 (complementation Group I). High doses of the mutant virus induced hydrocephalus in 87% of the mice. Infected mice were clinically asymptomatic, except for a few with transient hind-limb paralysis and proximal muscle weakness. After inoculation, mice were killed every other day for the first two weeks, and weekly thereafter for two months. Virological studies showed replication in the brain in the first nine days post-inoculation (DPI). Neutralizing antibody titers increased rapidly after 15 DPI, and elevated titers were measured at 30 DPI. Pathologically, there was patchy ependymal cell necrosis in the aqueduct and lateral ventricles, as early as the second DPI. Mild meningoencephalitis and severe ependymal cell necrosis with focal aqueductal stenosis were present iun the first two weeks of infection. Hydrocephalus began as early as 10 DPI and became severe at 28 DPI. This represents the first animal model for hydrocephalus following IC inoculation of a spontaneous ts mutant of a rhabdovirus. In our study, inoculation of mice with wild-type VSV and with other spontaneous and chemical ts mutants of VSV IC as well as with tspi 364 by other routes did not cause hydrocephalus. PMID- 6290613 TI - Direct afferent excitation and long-term potentiation of hippocampal interneurons. PMID- 6290614 TI - Effects of GABA on stimulus-evoked changes in [K+]o and parallel fiber excitability. PMID- 6290615 TI - Antagonism of GABA-mediated responses by d-tubocurarine in hippocampal neurons. PMID- 6290616 TI - Transmission at a central inhibitory synapse. I. Magnitude of unitary postsynaptic conductance change and kinetics of channel activation. PMID- 6290617 TI - Transmission at a central inhibitory synapse. III. Ultrastructure of physiologically identified and stained terminals. PMID- 6290618 TI - Short time scale correlations between spike activity of neighboring respiratory neurons of nucleus tractus solitarius. PMID- 6290619 TI - Associative neural and behavioral change in Hermissenda: consequences of nervous system orientation for light and pairing specificity. PMID- 6290620 TI - Stimulus and recovery dependence of cat cochlear nerve fiber spike discharge probability. PMID- 6290621 TI - Ultrastructural changes in rat peripheral nerve following pneumatic tourniquet compression. AB - The sciatic nerves of 12 male rats were examined in the electron microscope 14 days after pneumatic tourniquet compression. Tourniquet pressure was maintained at 300 mmHg for varied lengths of time (30 minutes to 3 hours). Nerves compressed for 30 minutes showed very mild fissuring of the myelin without axonal degeneration. Examination of nerves compressed for 1 to 3 hours showed progressively more varied and extensive damage. Changes included splaying of myelin lamellae, axonal shrinkage with periaxonal edema. Schwann cell hypertrophy, and an increase in the number of microtubules and mitochondria per unit area. The myelin sheaths of some fibers, compressed for more than 2 hours, were completely ruptured. These changes resemble nerve lesions which could be induced by a variety of experimental procedures. Ultrastructural changes produced by tourniquet compression are apparently time-related and affect large-diameter nerves more profoundly than smaller-diameter nerves. The data reported provide an explanation for delayed muscle rehabilitation experienced by patients who have undergone extremity surgery with pneumatic tourniquet application. The evidence presented suggests that the incidence of tourniquet palsy may be far greater than previously recognized. PMID- 6290622 TI - A noninvasive method for evaluating portal circulation by administration of Ti 201 per rectum. AB - A new method for evaluating portal systemic circulation by administration of TI 201 per rectum was performed in 13 control subjects and in 65 patients with various liver diseases. In normal controls, the liver was visualized on the 0--5 min image whereas the images of other organs such as the heart, spleen, and lungs were very poor. In patients with liver cirrhosis associated with portal-systemic shunt, and in many other patients with hepatocellular damage, the liver was not so clearly visualized, whereas radioactivity in other organs, especially the heart, became evident. The heart-to-liver uptake ratio at 20 min after administration (H/L ratio) was significantly higher in liver cirrhosis than in normals and patients with chronic hepatitis (p less than 0.001). The patients with esophageal varices showed a significantly higher H/L ratio compared with that in cirrhotic patients without esophageal varices (p less than 0.001). The H/L ratio also showed a significant difference (p less than 0.01) between Stage 1 and Stage 3 esophageal varices. Since there were many other patients with hepatocellular damage who had high H/L ratios similar to those in liver cirrhosis, the effect that hepatocellular damage has on the liver uptake of TI 201 is also considered. Our present data suggest that this noninvasive method seems to be useful in evaluating portal-to-systemic shunting. PMID- 6290624 TI - Relative biopotency of dietary ergocalciferol and cholecalciferol and the role of and requirement for vitamin D in rainbow trout (Salmo gairdneri). AB - A growth assay was conducted for six consecutive 28-day periods by using triplicate groups of 110 rainbow trout with an average initial body weight of 3.0 g. Ergocalciferol (vitamin D2 or D2) and cholecalciferol (vitamin D3 or D3) were included to provide levels of 200, 400 and 800 iu/kg in a semipurified casein, gelatin diet. Further treatments with 0 vitamin D and 1600 IU/kg of D3 were also included. The resulting growth curves were significant for parallelism. Statistical analysis showed that D3 was 3.27 times as potent as D2 (limits 2.33 to 4.58). The dietary requirement for D3 was found to be in excess of 800 iu/kg of diet. Vitamin D-deficient fish showed no change in bone ash but exhibited clinical manifestations of tetany with no hypocalcemia. A complete absence of tetany was seen only in the groups fed 800 and 1600 IU of D3 per kilogram. None of the levels of D2 used were sufficient to completely alleviate symptoms of this disorder. These studies of rainbow trout provide evidence that vitamin D is required for the normal functioning of white muscle without altering the calcium content of the plasma or epaxial musculature. PMID- 6290623 TI - Graves' disease: thyroid function and immunologic activity. AB - Patients with Graves' disease were studied for two years during and after a twelve-month course of treatment. Disease activity was determined by repeated measurements of thyroidal uptake of [99mTc]pertechnetate during tri-iodothyronine administration. These in-vivo measurements of thyroid stimulation were compared with the results of in-vitro assays of Graves, immunoglobulin (TSH binding inhibitory activity--TBIA). There was no correlation between the thyroid uptake and TBIA on diagnosis. Pertechnetate uptake and TBIA both declined during the twelve months of antithyroid therapy. TBIA was detectable in sera from 19 of the 27 patients at diagnosis; in 11 of these 19 patients there was a good correlation (p less than 0.05) throughout the course of their disease between the laboratory assay of the Graves, immunoglobulin and the thyroid uptake. Probability of recurrence can be assessed but sustained remission of Graves' disease after treatment cannot be predicted from either measurement alone or in combination. PMID- 6290625 TI - The biological activity of 25-hydroxycholecalciferol and 1,25- dihydroxycholecalciferol for rainbow trout (Salmo gairdneri). AB - Dietary 25-hydroxycholecalciferol (25-hydroxycholecalciferol (25-OH-D3) and 1,25 dihydroxycholecalciferol [1,25-(OH)2D3] showed vitamin D activity in rainbow trout. However, inclusion of dietary cholecalciferol (vitamin D3 or D3), ergocalciferol (vitamin D2 or D2), 25-OH-D3 or 1,25-(OH)2D3 did not result in the presence of detectable levels of vitamin D or 25-OH-D in the blood plasma of the fish. Fish fed the diet devoid of vitamin D over an extended period of time showed symptoms of a droopy-tail or "lordosis-like" syndrome that appeared to be related to muscle weakness since x-ray examination indicated no abnormality in vertebral development. The requirement for vitamin D as cholecalciferol was in excess of 1600 IU/kg diet and may be as high as, or higher than 2400 IU/kg diet. PMID- 6290626 TI - Effect of pectin structure on protein utilization by growing rats. AB - The effect of pectins' molecular weight (MW) and degree of esterification (DE) on apparent digestibility (D), net protein utilization (NPU) and biological value (BV) of casein were tested. The pectins prepared differed in MW and DE respectively as follows: P-1 (180,000, 73%); P-2 (300,000, 28%); P-3 (60,000, 74%) and P-4 (75,000, 40%) and were fed as 10% of a purified diet containing 10% casein to growing male rats for 10 days. All preparations decreased D, with P-2 and P-4 showing milder effects than P-1 or P-3. Increased NPU was observed with pectins of low NW and/or DE, being significant only with P-4. Diets containing P 2, P-3 or P-4 had higher BV for casein than the pectin-free diet. Feeding these pectins in a protein-free diet did not cause a significant change in the rats' body N content. Thus, the increase in the BV of casein was attributed to improvements in the pattern of amino acids absorbed by rats fed pectins with low MW and/or low DE. PMID- 6290627 TI - Effects of wheat bran on the metabolism of calcium-45 and zinc-65 in rats. AB - Metabolic balances of orally administered tracer doses of 45Ca and 65Zn were carried out in 20 weanling male rats to examine the effect of different levels of wheat bran (0.5 10 and 15%) on the bioavailability of these minerals in diets containing respectively 9.32, 9.26, 9.28 and 10.76 mg/g of calcium and 58, 63, 68, and 76 ppm of zinc. To estimate the influence of adaptation to feed consumption, the experiment was repeated after a 10-day period of adaptation; a split-plot-in-time design was used for statistical analysis. The growth rate was not affected by bran administration. No significant change was found in 45Ca or 65Zn absorption due to the addition of wheat bran to the diets. The results in the two repetitions were the same. PMID- 6290628 TI - Isolation of sterically 'hindered' 7-cis-beta-carotene. PMID- 6290629 TI - Primary central adenoid cystic carcinoma of the mandible. PMID- 6290630 TI - Metastatic hepatocellular carcinoma of the mandible. PMID- 6290631 TI - A new liquid medium for xenic cultivation of Entamoeba histolytica and other lumen-dwelling protozoa. PMID- 6290632 TI - Effect of taurine on the gastric absorption of drugs: comparative studies with sodium lauryl sulfate. AB - In order to clarify the mechanism of taurine-induced enhancement of drug absorption, some comparison was made between the actions of taurine and sodium lauryl sulfate (SLS), an anionic surfactant, by using the loop of rat stomach in situ. SLS enhanced the gastric absorption of aspirin and salicylamide, and the SLS-induced enhancement seems to be Na+-dependent. SLS failed to enhance the absorption of both drugs when NaCl was completely replaced by mannitoL or in the presence of ouabain even in the NaCL medium. In addition, even at the lower concentration of 0.05 mM, SLS enhanced the absorption of the two drugs, the extent of which is nearly corresponding to those of taurine. The actions of SLS and taurine on drug absorption are considered to have some similar points. The mechanism of the action was discussed in relation to the cyclic AMP system. PMID- 6290633 TI - Renal tubular effects of sodium fluoride. AB - Administration of sodium fluoride results in vasopressin-resistant polyuric "renal failure" resembling nephrogenic diabetes insipidus. However, the renal tubular site of action of fluoride is not clear. Fischer 344 rats received acute i.v. infusions of sodium fluoride (0.3, 1.47 and 2.20 mumol/min/kg b.wt.) for 2.5 hr which resulted in dissipation of the renal medullary tissue osmotic gradient and a sustained, dose-related increase in fractional sodium excretion and urine flow. In additional experiments, free water reabsorption and excretion were decreased by fluoride, but the decrease in free water excretion occurred only when the fluoride-induced polyuria preceded the onset of the water diuresis. Slices of renal medulla from fluoride-treated rats had lower cyclic AMP concentrations than did slices from control rats and the responsiveness of the medullary tissue to vasopressin was markedly reduced. These data indicate that the fluoride ion dissipates the concentration gradient in the renal medulla largely by inhibiting NaCl reabsorption in the ascending limb of Henle's loop and inhibits antidiuretic hormone-mediated water reabsorption across the collecting duct. PMID- 6290634 TI - Evidence for sigma opioid receptor: binding of [3H]SKF-10047 to etorphine inaccessible sites in guinea-pig brain. AB - A portion of the specific binding of tritiated SKF-10047 to the guinea-pig brain suspension of the particulate fraction is not inhibited by the strong narcotic analgesic l-etorphine. The binding properties of these etorphine-inaccessible (EI) sites were examined. The specific binding of [3H]SKF-10047 to the EI sites is saturable. Scatchard analysis of the saturation curve revealed a single class of binding sites with apparent Kd of 252 nM and an estimated Bmax of 663 fmol/mg of protein. The EI binding was reduced by heat treatment, trypsin digestion and phospholipase C digestion. The presence of sodium ions slightly increased specific EI binding. Lithium ion increased the EI binding by about 38% at the optimal concentration of 1 mM. Divalent cations such as Mg++, Ca++ and Mn++ reduced EI binding. Morphine-like drugs such as morphine, levorphanol and naltrexone were poor inhibitors for the EI binding, whereas opioid derivatives such as pentazocine, dextrallorphan, cyclazocine, SKF-10047 and dextrorphan were potent inhibitors. Nonopioid drugs such as haloperidol, imipramine, pimozide and propranolol were also potent inhibitors of the EI binding. Distribution of the EI sites in brain was different from that of the mu receptor: highest concentration of EI sites was found in brainstem, midbrain and cerebellum, whereas lower concentrations were found in striatum and cortex. It is suggested that EI sites are not mu receptors but may represent sigma receptors in the central nervous system, mediating psychotomimetic effects of several opioids and other drugs. PMID- 6290635 TI - Baclofen selectively inhibits transmission at synapses made by axons of CA3 pyramidal cells in the hippocampal slice. AB - The effects of baclofen, an antispastic drug, on excitatory transmission were tested by bath application to the hippocampal slice preparation. (+/-)-Baclofen (20 microM) strongly depressed extracellularly recorded synaptic responses to stimulation of projections that originate from CA3 hippocampal pyramidal cells. Responses to stimulation of four other excitatory pathways were little affected and the amplitudes of presynaptic fiber potentials and antidromic responses were unaltered. When tested on the Schaffer collateral-commissural-CA1 pyramidal cell synapse. (-)-baclofen depressed the amplitude of the extracellular excitatory postsynaptic potential with an IC50 of 3.7 microM and was 180 times more potent than (+)-baclofen. gamma-Aminobutyric acid, 3-aminopropanesulfonic acid and imidazole-4-acetic acid also inhibited transmission at this site. Baclofen could suppress the response completely, and its action was unaffected by bicuculline. In contrast, imidazole-4-acetic acid could suppress the response by a maximum of only 75%, and its action was highly sensitive to bicuculline. gamma-Aminobutyric acid and 3-aminopropanesulfonic acid could suppress the response completely, and their actions were relatively weakly antagonized by bicuculline. These results are consistent with the hypothesis that baclofen inhibits excitatory transmission by interacting with a bicuculline-insensitive gamma-aminobutyric acid receptor. These receptors may be located on one type of glutamatergic/aspartergic synaptic terminal, exemplified by axon terminals of CA3 hippocampal pyramidal cells. Synapses made by these axons may therefore serve as models for studying the mechanism of action of baclofen. PMID- 6290636 TI - Effects of phenoxybenzamine or N-methyl chlorpromazine on regional cerebral blood flow: comparison of central and peripheral alpha adrenergic receptor antagonism. AB - A comparison of the central and peripheral effects of alpha adrenergic receptor antagonism on regional cerebral blood flow was examined utilizing N-methyl chlorpromazine, an alpha adrenergic receptor antagonist which does not cross the blood-brain barrier, or phenoxybenzamine, an alpha adrenergic receptor antagonist which does enter the brain when injected systemically. Regional cerebral blood flow was monitored in 17 brain regions of 16 pentobarbital-anesthetized New Zealand White rabbits using radioactively tagged microspheres (15 +/- 3 micrometer in diameter). There was a significant overall difference in regional cerebral blood flow distribution in the control state. Although phenoxybenzamine had no significant effect on average global cerebral blood flow, this agent significantly redistributed flow among the various regions. Flow was significantly decreased in some cortical regions, the hippocampus and cerebellum, whereas flow was increased in the pons and substantia nigra. There were no significant effects of N-methyl chloropromazine on average global cerebral blood flow or its regional distribution. Central alpha adrenergic receptor blockade probably decreased regional cerebral blood flow in brain regions rich in neuronal alpha adrenoreceptors by decreasing regional metabolic and/or neuronal activity, while increasing metabolic and/or neuronal activity in the pons and substantia nigra via a positive feedback mechanism causing an increased flow rate in these regions. Peripheral alpha adrenergic receptor blockade appears to have little or no effect on regional cerebral blood flow. PMID- 6290639 TI - Facilitation of ganglionic transmission by sulpiride: evidence for an inhibitory role of dopamine in the canine sympathetic ganglion. AB - Effects of the (R) and (S) enantiomers of sulpiride, a potent dopamine (DA) antagonist, on ganglionic transmission were studied in anesthetized dogs. The pre and postganglionic nerves of cardiac sympathetic ganglia were stimulated electrically, and heart rate was monitored as a measure of ganglionic transmission and sympathetic nerve activity. The heart rate was free from influence of the central nervous system. (R)- And (S)-sulpiride injected i.a. close to the blood supply of the ganglia did not alter basal heart rate, but facilitated ganglionic transmission as demonstrated by an increase in the tachycardia induced by preganglionic nerve stimulation. The (R) enantiomer was 4 times more active than the (S) enantiomer in this respect. Neither enantiomer affected the tachycardia induced by postganglionic nerve stimulation. Norepinephrine and DA injected i.a. caused inhibition of the tachycardia induced by preganglionic nerve stimulation. The inhibitory effect of both catecholamines was antagonized by the sulpiride enantiomers (R)-sulpiride was about 4-fold more potent than (S)-sulpiride in antagonizing DA, whereas (S)-sulpiride was more active against norepinephrine. The sulpiride enantiomers affected neither the tachycardia induced by i.a. administration of acetylcholine nor the bradycardia induced by vagal nerve stimulation. Thus, cholinesterase inhibition and ganglionic stimulation were excluded. These data are, therefore, consistent with the hypothesis that the facilitatory action of the sulpiride enantiomers is related to the antagonism of catecholamines. Positive correlation between the activity of the (R) enantiomer to facilitate ganglionic transmission and to antagonize DA suggests that DA is a physiologically released catecholamine modulating transmission in the cardiac sympathetic ganglia of the dog. PMID- 6290637 TI - Effect of reserpine pretreatment on mechanical responsiveness and [125I]Iodohydroxybenzylpindolol binding sites in the guinea-pig right atrium. AB - The inotropic and chronotropic responses of the guinea-pig right atrium to several pharmacologic agents were measured after acute (0.1 mg/kg/day x 1) and chronic (0.1 mg/kg/day x 7) reserpine administration. A small increase in the sensitivity of the pacemaker to isoproterenol occurred after acute reserpine treatment which was followed by a much greater change in sensitivity to the beta agonist when pretreatment was extended for 7 days. Chronotropic responsiveness to calcium, histamine and pilocarpine was not altered by reserpine pretreatment. The acute administration of reserpine resulted in a slight inotropic supersensitivity of paced right atria to isoproterenol, calcium and histamine. Pretreatment for 7 days produced an additional increase in inotropic sensitivity to isoproterenol but did not affect contractile responses to the other agents. The catecholamine specific nature of the supersensitivity induced by chronic reserpine treatment suggested that a change in the number and/or affinity of beta adrenergic receptors was involved. The radiolabeled beta adrenoceptor antagonist [125]iodohydroxybenzylpindolol (I-HYP) was used to test this hypothesis. Preliminary experiments revealed the presence of a single class of noninteracting (nH = 0.99), high affinity (Kd = 100 pM) binding sites which exhibited stereospecificity and saturability (47.2 fmol/mg of protein). The agonist potency series for the inhibition of I-HYP binding was identical to the series for mediating mechanical responses. Taken collectively this information suggests that the high affinity I-HYP binding site in the guinea-pig right atrium represents the beta adrenergic receptor. As determined by Scatchard analyses, neither acute (1-day) nor chronic (7-day) low-dose (0.1 mg/kg/day) reserpine administration altered the number or affinity of I-HYP binding sites. It is concluded that changes in beta receptor characteristics are not responsible for reserpine induced supersensitivity in this tissue. PMID- 6290638 TI - Bronchodilator mechanisms in bullfrog lung: differences in response to isoproterenol, theophylline and papaverine. AB - The effects of bronchodilators and smooth muscle relaxants on mechanical responses and lung cyclic nucleotide levels in the isolated hemilung of Rana catesbeiana demonstrate striking differences in intensity and time course of drug action in an unstimulated preparation of airway smooth muscle. Isoproterenol, nitroprusside and nitroglycerin elicit a fast onset relaxation (minutes) with ceiling effects at 20, 22 and 43%, respectively, of maximal relaxation. Theophylline, dibutyryl cyclic AMP and papaverine produce maximal or near maximal relaxation, but require 8 to 32 hr for peak effect. Papaverine-induced relaxation is accompanied by a slow increase in lung cyclic AMP and cyclic GMP and is markedly accelerated by isoproterenol. Theophylline (10(-3) M) produces no change in cyclic nucleotide levels and its relaxant effect is not accelerated by isoproterenol. The hierarchy of relaxant responses suggests drug action at discrete loci in a highly compartmentalized effector chain, with cyclic AMP dependent mechanisms separable into at least two components. The first is activated by isoproterenol and elicits a rapid, limited response, presumably reflecting an increase in cyclic AMP in a relatively restricted pool. The second is activated by papaverine and elicits a very slow, but complete relaxation, presumably reflecting a more pervasive or diffuse accumulation of cyclic AMP secondary to phosphodiesterase inhibition. The major portion of theophylline induced relaxation in this preparation appears to be independent of changes in cyclic nucleotide metabolism. PMID- 6290640 TI - Enhancement of cardiac actions of ouabain and its binding to Na+, K+-adenosine triphosphatase by increased sodium influx in isolated guinea-pig heart. AB - The rate of development of the positive inotropic action of ouabain is enhanced when the heart is stimulated at higher frequencies. A hypothesis that this enhancement is due to a stimulation of the glycoside binding to sarcolemmal Na+,K+-adenosine triphosphatase (ATPase) caused by an increase in intracellular Na+ available to the sodium pump was tested in isolated left atrial muscle preparations of guinea-pig heart, incubated at 30 degrees C and electrically stimulated at 0.5, 1 or 2 Hz. The rate of development of the positive inotropic action of ouabain was dependent on the frequency of stimulation. Each preparation was homogenized at a predetermined time and the fractional occupancy of Na+,K+ ATPase by ouabain was estimated from the decrease in the initial velocity of ATP dependent [3H]ouabain binding reaction. A parallel relationship was observed between effects of stimulation frequency of the positive inotropic action and those on the occupancy of Na+,K+-ATPase by ouabain. In quiescent preparations, a sodium ionophore, monensin, enhanced the development of contracture caused by a toxic concentration of ouabain and also the glycoside binding to Na+,K+-ATPase. Similar effects on the ouabain-induced contracture and on the glycoside binding were observed with either grayanotoxin I or batrachotoxin, agents known to increase sodium influx, when muscle preparations were exposed to these agents under 1.5 Hz stimulation and were subsequently tested for the actions of ouabain in quiescence. When the exposure to ouabain and either grayanotoxin I or batrachotoxin was restricted to quiescent period, the development of ouabain induced contracture and glycoside binding to Na+,K+-ATPase were not significantly altered. Monensin, grayanotoxin I or batrachotoxin failed to significantly affect [3H]ouabain binding to muscle homogenates when added to the medium for the labeled glycoside binding assay. These results indicate that intracellular sodium ions promote the ouabain binding to Na+,K+-ATPase and thereby enhance the development of glycoside actions in the isolated atrial muscle of guinea-pig heart. The "beat-dependent" onset of the glycoside action is at least partially explained from the effect of membrane depolarization to increase Na+ available to the sodium pump and to enhance the glycoside binding. PMID- 6290641 TI - Operational characteristics of the inhibitory feedback mechanism for regulation of dopamine release via presynaptic receptors. PMID- 6290642 TI - Cannabimimetic activity from CP-47,497, a derivative of 3-phenylcyclohexanol. AB - CP-47,497 (cis-3-[2-hydroxy-4(1,1-dimethylheptyl)phenyl]-cyclohexan-1-ol) is characterized as a cannabimimetic agent, with 3 to 28 times greater potency than delta 9-tetrahydrocannabinol delta 9-THC), based on the following. In common with delta 9-THC and other active structures closely related chemically to delta 9 THC, CP-47,497 exerts analgesic, motor depressant, anticonvulsant and hypothermic effects. It elicits vocalization in palpated rats and ataxia in dogs. In drug discrimination studies in rats, the stimulus properties of delta 9-THC (3.2 mg/kg i.p.) are generalized to CP-47,497, with an absolute threshold dose 3 to 14 times lower than the threshold dose of delta 9-THC itself, depending on route. Furthermore, rats are unable to discriminate between the stimulus properties of equated i.p. doses of delta 9-THC and CP-47,497 after prolonged training. Despite its potent behavioral effects, CP-47,497, like delta 9-THC, does not resemble standard antipsychotic, antidepressant, antianxiety or hypnotic drugs in simple drug interaction tests. Based on its pharmacology, CP-47,497 exemplifies a simplified structure capable of producing many effects in common with those of delta 9-THC and 9-normethyl-9 beta-OH-hexahydrocannabinol. PMID- 6290643 TI - Effect of chronic ethanol treatment on temperature dependence and on norepinephrine sensitization of rat brain (Na+ + K+)-adenosine triphosphatase. AB - Rat brain (Na+ + K+)-adenosine triphosphatase is inhibited by ethanol (EtOH) in vitro, the inhibition being greater in the presence of norepinephrine (NE). Enzyme preparations from EtOH-tolerant rats show less inhibition by EtOH in vitro and less sensitization by NE. To investigate the mechanism of these changes, the enzyme activity of brain microsomes from tolerant and sucrose-control rats was measured at temperatures from 10-40 degrees C. Preparations from nonwithdrawn and 24-hr withdrawn rats were studied in the absence of in vitro additions, and in the presence of 1 microM NE, 50 mM EtOH or 440 mM EtOH separately, and of 1 microM NE + 50 mM EtOH. From Arrhenius plots of the results, the transition temperature (Td) was calculated by a method of successive approximations, and the activation energies were calculated from the segments above and below Td. Chronic EtOH treatment significantly decreased Td, but increased activation energies below Td. These findings suggest different effects on membrane matrix lipids than on boundary lipids adjacent to the enzyme. However, EtOH-tolerant preparations showed less effect of EtOH in vitro than did control preparations, on both Td and activation energies. Preparations from EtOH-tolerant and withdrawn rats behaved almost identically, indicating that the changes accompany tolerance and are not withdrawal effects. NE + 50 mM EtOH produced the same effects as 440 mM EtOH alone, in all preparations. EtOH tolerance reduced the sensitizing effect of NE. EtOH is interpreted as affecting both the boundary lipids and the apoenzyme itself. PMID- 6290644 TI - Apparent frequency-dependent effect of clonidine on cardiac sympathetic transmission: the role of neuronal uptake. AB - Decreases in spontaneous atrial period (tachycardia) were measured in atropinized guinea-pig isolated right atria in response to intramural sympathetic nerve stimulation. Electrical field pulses were applied only during the atrial refractory period to avoid arrhythmias. Responses to one to six field pulses delivered in a single refractory period were substantially reduced by clonidine (1-1000 nM), providing no evidence that the blocking action of this drug is frequency-dependent. Stimulus-response lines to one, two or four field pulses delivered as one field pulse per refractory period (1/1) were displaced to the right by clonidine (0.1 microM) such that 8, 16 and 32 field pulses were required to obtain responses equivalent to control. Stimulus-response lines to lower frequencies of field stimulation (one field pulse every 4 or 8 refractory periods) were lower in slope and displaced further by clonidine, whereas lines from stimuli at higher frequencies (2/1 and 4/1) were steeper and less displaced by clonidine. These findings confirmed an "apparent" frequency-dependent blocking action of clonidine. However, substantial blockade of neuronal uptake by desipramine (0.1 microM) raised the slope and decreased the shift of the stimulus response lines to the lower frequencies of field stimulation compared with 2/1 or 4/1. Clonidine (0.1 microM) had no effect on the responses to exogenous norepinephrine. We suggest that clonidine reduces the amount of norepinephrine released per field pulse independently of the frequency of stimulation. The decrease in tissue response may appear to be frequency-dependent under some circumstances due to the effects of uptake and removal processes. PMID- 6290645 TI - Kinetics of the sodium pump in the frog choroid plexus. AB - 1. Active Na+ transport across the choroid plexus is mediated by Na-K pumps in the brush border membrane of the epithelium. We have studied the kinetics of Na+ pumping across the brush border membrane using a 22Na efflux procedure. 2. Frog choroid plexuses were loaded with 22Na in K+ -free, cold Ringer solution containing various Na+ concentrations. The efflux of 22Na from the tissues was monitored at 22 degrees C and the rate constant (k) was estimated for each flux interval. The pump component was obtained from the change in k(delta k) induced by the addition of KCl to the superfusate. 3. Ouabain blocked this K+ -sensitive Na+ efflux and other monovalent cations substituted for K+ in the sequence Tl+, Rb+, K+, greater than NH+4 greater than Cs+ much greater than Li+. Active sodium transport (delta k x Nac) increased in a sigmoidal and saturable way with intracellular Na+ and with extracellular K+ concentrations. 4. Kinetic parameters were estimated using the model of Garay & Garrahan (1973). The data indicate that there were two ligand binding sites for K+ on the outside face and three ligand binding sites for Na+ on the cytoplasmic face of the pump. The Kms for potassium and sodium were 1 . 1 and 7 mM. 5. The results also suggest that the Na-K pump has a coupling ratio of 1 . 5. We conclude that the Na-K pump in this epithelium is very similar to those in single cells such as erythrocytes, nerve and muscle. PMID- 6290646 TI - Passive rubidium fluxes mediated by Na-K-ATPase reconstituted into phospholipid vesicles when ATP- and phosphate-free. AB - 1. Phospholipid vesicles reconstituted with Na-K-ATPase from pig kidney, show slow passive pump-mediated (86)Rb fluxes in the complete absence of ATP and phosphate.2. The Rb fluxes are inhibited in vesicles prepared from enzyme pre treated with either ouabain or vanadate ions. Rb fluxes through Na-K pumps oriented inside-out or right-side out by comparison with the normal cellular orientation can be distinguished by effects of vanadate on one or both sides of the vesicle.3. (86)Rb uptake into Rb-loaded vesicles represents a (86)Rb-Rb exchange. The maximal rate of exchange through inside-out and right-side out oriented pumps is equal, suggesting a random arrangement of the pumps across the vesicle membrane. This Rb-Rb exchange is half-saturated on inside-out and right side out pumps at about 0.6 and 0.2 mM-external Rb respectively.4. (86)Rb uptake into Rb-free vesicles represents a net Rb flux. The Rb uptake through inside-out pumps has a maximal rate about equal to the Rb-Rb exchange, half-saturates at an external Rb concentration of roughly 0.5 mM, and shows evidence for co operativity. Net Rb uptake through right-side out pumps is very slow, and half saturates at roughly 0.1 mM external Rb.5. K ions at low concentrations in the exterior medium stimulate (86)Rb uptake, but at high concentrations, inhibit. Na ions in the exterior medium always inhibit (86)Rb uptake. The result suggests that K ions are transported in co-operative fashion together with Rb ions, while Na ions block the Rb fluxes.6. The presence of Rb congeners at the vesicle interior raises the (86)Rb uptake through inside-out pumps with the decreasing order of effectiveness: Li > Na > Cs > K > Rb. Stimulation by Na ions involves a Rb-Na exchange.7. Turnover numbers were estimated from parallel measurement of Na/K pump mediated fluxes and amount of covalent phosphoenzyme. In units of moles of ion per mole of phosphoenzyme per second at 20 degrees C the following values were obtained: ATP-dependent Na-Rb exchange, 43; (ATP+phosphate)-stimulated Rb-Rb exchange, 7. For (ATP+phosphate)-independent fluxes: Rb-Rb exchange 0.25; net Rb uptake 0.15 and Rb-Na exchange 0.65.8. Mg ions in the exterior medium inhibited both net and exchange Rb fluxes through inside-out pumps in a manner antagonistic with respect to Rb. Mg and vanadate ions inhibit the Rb fluxes in a synergistic fashion.9. The results are interpreted in terms of a model in which net and exchange (86)Rb fluxes occur via conformational transitions between form E(1) which binds Rb at the cytoplasmic face of the protein, the form E(2) (Rb)(occ) containing occluded Rb ions and a form E(2) which binds Rb at the extracellular face of the protein. A kinetic analysis allows us to identify rate-limiting steps of the transport cycle by making use of our transport data in combination with values of rate-constants for conformational transitions observed directly in isolated Na-K-ATPase. PMID- 6290647 TI - Effects of atp or phosphate on passive rubidium fluxes mediated by Na-K-ATPase reconstituted into phospholipid vesicles. AB - 1. The passive Rb fluxes mediated by the Na-K pump in reconstituted vesicles, described by Karlish & Stein (1982), are affected by ATP or by phosphate acting separately.2. Rb-Rb exchange through inside-out pumps is stimulated by ATP at low concentrations and is inhibited at high concentrations. There are mutual effects of Rb at cytoplasmic sites and ATP. The higher is the Rb concentration, the greater is the degree of stimulation and the less is the inhibition of exchange by ATP, and the higher are the concentrations of ATP required to produce effects. ATP stimulates Rb-Rb exchange maximally by about 5-fold.3. There are similar effects of ATP on zero-trans net Rb uptake through inside-out pumps. However, much lower degrees of stimulation and greater inhibition of the net flux by ATP are observed, and much lower concentrations of ATP are required for these effects, by comparison with those on Rb-Rb exchange.4. Rb uptake on inside-out pumps in Na-loaded vesicles shows only inhibition by ATP.5. Phosphate effects require the presence of Mg(0) ions. At low Mg(0) concentrations (up to 100 muM) phosphate moderately stimulates Rb uptake into Rb-free or Rb-loaded vesicles (about 50%), but has no effect on Rb uptake into Na-loaded vesicles. At millimolar concentrations of Mg(0) ions, phosphate strongly inhibits the Rb uptake into Rb-free or Na-loaded vesicles but has no effect on Rb uptake into Rb loaded vesicles.6. The separate effects of ATP and of phosphate are explained in terms of the model proposed by Karlish & Stein (1982), modified to take into account stimulation of the conformational transition E(2)(Rb)(occ) --> E(1) Rb by ATP, and stimulation of the conformational transition E(2)(Rb)(occ) --> E(2) Rb by phosphate due to phosphorylation of the protein. PMID- 6290648 TI - Combined effects of ATP and phosphate on rubidium exchange mediated by Na-K ATPase reconstituted into phospholipid vesicles. AB - 1. Phospholipid vesicles reconstituted with Na-K-ATPase show an (ATP+phosphate) stimulated Rb-Rb exchange, with properties similar to the K-K exchange of human red cells. This includes a rate 15-20% of the rate of active ATP-dependent Na-K exchange.2. We have studied activation of this Rb-Rb exchange by ATP at fixed phosphate concentrations and by phosphate at fixed ATP concentrations. It is found for both ATP and phosphate that with low concentrations of the fixed ligand an increase in concentration of the complementary ligand produces first stimulation and then inhibition of Rb-Rb exchange. At high concentrations of the fixed ligand the complementary ligand shows only saturation behaviour.3. The pattern of activation and of inhibition by ATP and by phosphate is affected by the Rb(0) concentration in the exterior medium, in that higher concentrations of Rb(0) counteract inhibitory effects of high concentrations of ATP and phosphate.4. (ATP+phosphate)-stimulated Rb-Rb exchange is activated by Rb(0) in the exterior medium along a sigmoid curve. An increase of Rb(i) within the vesicles, which raises the maximal velocity of Rb-Rb exchange, is accompanied by a smaller increase in the Rb(0) concentration required for half-maximal stimulation of the Rb-Rb exchange.5. The data are interpreted in terms of a model similar to those proposed by Karlish & Stein (1982a,b), but extended to include simultaneous effects of ATP and phosphate. Inhibitions by high concentration of ATP or phosphate arise as a result of stabilization of E(1) ATP or E(2)-P forms respectively, in the presence of low concentrations of the complementary ligand. With high concentrations of the fixed ligand, saturation behaviour of the varying ligand is observed because the occluded Rb forms become the dominant transport intermediates. The occluded Rb forms bind both ATP and phosphate weakly and independently. The effects of ATP together with phosphate are accounted for by a simple combination of their separate effects on the Rb-Rb exchange.6. We suggest that the functional role of the occluded Rb form E(2) (Rb)(occ) in active transport is to minimize passive cation leaks through the system and allow control of the direction of cation movements by binding of physiological ligands such as ATP or phosphate. PMID- 6290649 TI - On the nature of histamine-mediated slow hyperpolarizing synaptic potentials in identified molluscan neurones. AB - 1. Standard intracellular stimulating and recording techniques were used to test the correspondence between monosynaptic post-synaptic potentials (p.s.p.s) evoked by histamine-containing C-2 neurones and responses to focally applied histamine recorded from two classes of identified post-synaptic neurones in the cerebral ganglion of Aplysia californica.2. Two types of p.s.p.s were examined: (1) a monophasic slow hyperpolarizing potential (I(s)p.s.p.) lasting 1-2 sec; and (2) a biphasic p.s.p. consisting of a fast depolarizing component <0.5 sec in duration (E(f)p.s.p.) plus a slow hyperpolarizing potential (I(s)p.s.p.) designated the E(f)I(s)p.s.p.3. Ionophoretic or pressure applied histamine mimicked both p.s.p.s and produced conductance increases in the post-synaptic neurones similar to those associated with the evoked p.s.p.s.4. The reversal potentials (E(rev)) for the I(s)p.s.p. and E(f)I(s)p.s.p., estimated by extrapolation, were -85+/-5.3, -35+/ 5.5, and -83+/-8.1 mV (mean+/-S.D.), respectively. The I(s)p.s.p.s were produced by an increase in potassium conductance because their E(rev)s were shifted about 16 mV by doubling or halving the concentration of extracellular potassium and they could be eliminated by cooling or by intracellular injection of TEA ions.5. The average E(rev) values for the slow hyperpolarizing histamine responses were similar to those for the I(s)p.s.p.s; about -83 and -86 mV in neurones receiving the monophasic I(s)p.s.p.s and biphasic E(f)I(s)p.s.p., respectively.6. Cimetidine, an antihistamine drug that selectively blocks histamine receptors associated with potassium conductances in Aplysia, reversibly abolished the I(s)p.s.p.s and slow hyperpolarizing responses to focally applied histamine. In similar concentrations, cimetidine had no discernible effects on the E(f)p.s.p. and depolarizing response to histamine or on several different types of p.s.p.s mediated by the C-2 neurones.7. It is proposed that the I(s)p.s.p.s are mediated by histamine released from the C-2 neurones. PMID- 6290650 TI - Ungulate cardiac purkinje fibres: the influence of intracellular pH on the electrical cell-to-cell coupling. AB - 1. Passive electrical properties of sheep cardiac Purkinje fibres were assessed by performing linear cable analysis. In a separate set of experiments pH(i) was monitored using recessed-tip pH-sensitive micro-electrodes.2. In Tris-buffered Tyrode solution (nominally CO(2)-free), the pH(i) was 7.27, in bicarbonate buffered solution equilibrated with 6% CO(2), the mean pH(i) was 7.02.3. Application of 15 mM-NH(4)Cl produced a rapid intracellular alkalinization (0.19 pH units), followed by a slower acidification. Removal of NH(4)Cl gave rise to a slow and transient intracellular acidification (0.5 pH units).4. The biphasic and transient shift in pH(i), induced by the NH(4)Cl treatment, was accompanied by a change of the inside longitudinal resistance per unit fibre length, r(i), displaying a similar time course. The increase in pH(i) produced a maximum decrease in r(i) of 16.4%, while the decrease in pH(i) yielded a maximum increase in r(i) of 30.4%.5. Changing from bicarbonate-buffered Tyrode solution equilibrated with 6% CO(2) to Tris-buffered Tyrode solution led to an increase in pH(i) (0.26 pH units). A subsequent change to bicarbonate-buffered Tyrode solution equilibrated with 15% CO(2) produced a decrease in pH(i) (0.48 pH units). Both changes were sustained.6. This CO(2) protocol gave rise to corresponding changes in r(i); the intracellular alkalosis was associated with a decrease in r(i) (21.2%), and the intracellular acidosis was accompanied by an increase in r(i) (30%).7. Based on recent findings showing an interaction between pH(i) and pCa(i) (Hess & Weingart, 1980), it is concluded that the changes in r(i) are directly caused by protons and not indirectly via secondary changes of the [Ca(2+)](i).8. The pH(i)-dependent changes in r(i) are likely to reflect alterations of the nexal resistance, r(n), because the cytoplasmic resistance, r(c), has the inverse sensitivity to pH(i).9. Unlike pCa(i), pH(i) would seem to be able to modify the cell-to-cell coupling by increasing or decreasing r(i) over a rather narrow range, without ever producing electrical uncoupling.10. Because of basic differences in the action of calcium and protons on the cell-to-cell coupling (magnitude of the effect, operative concentration range), it is tempting to conclude that there is more than one kind of binding site which controls the nexal conductance. PMID- 6290651 TI - [Physiology of nociception]. PMID- 6290652 TI - [Corticohypophysiotropic and corticotropic activity in adrenalectomized male and female rats after psychic stress]. AB - We show modifications in the hypothalamic CRF activity and plasma ACTH concentration in adult rats of both sexes, which were five day sham-operated or adrenalectomized and killed either under basal conditions or after a 3 min period or psychological stress. 1. Under basal conditions, the inhibition of the basal release of ACTH is suppressed in 5 day adrenalectomized rats and a sex difference appears: plasma ACTH concentration is twice as great in females than in males. 2. After a 3 min period of psychological stress, the usual increase in hypothalamic CRF activity observed in sham-operated rats, which seems to be sex-related, does not appear in adrenalectomized male or female rats. However, in adrenalectomized rats, the maximal increase in plasma ACTH concentration occurred more rapidly, with a rate 10 times as great in males and 4 times as great in females, than in sham-operated rats. Differences between the sexes in the maximal increase in plasma ACTH concentration remains 1,6 times as great in females than in males. 3. Our results confirm that corticosteroids exert: (1) a tonic feedback inhibition of the basal release of ACTH, (2) a fast feedback inhibition of the stress induced release of ACTH; the promote an increase in the hypothalamic CRF content. Relative intensity of these two inhibitory mechanisms seems to be sex-related. PMID- 6290653 TI - ACTH and plasma corticosteroid levels in young calves. AB - 1. Plasma sodium, potassium, aldosterone and cortisol levels were measured in seven 11-day-old calves after ACTH infusion and in five control animals. 2. During the 24 h following ACTH infusion the concentration of sodium and potassium in plasma and urine showed no significant variation either in treated or in control calves. 3. ACTH infusion induced a significant increase in plasma cortisol levels, which did not appear in control calves. Plasma aldosterone levels did not vary significantly in treated or in control calves. PMID- 6290654 TI - Multicentric glomus tumour of the stomach (a case report). PMID- 6290655 TI - Conditions favoring gametocytogenesis in the continuous culture of Plasmodium falciparum. PMID- 6290656 TI - Characterization of anti oLH beta-antibodies acting as contraceptives in rhesus monkeys. I. In vitro binding properties. AB - Female monkeys actively immunized with oLHbeta are infertile when circulation antibody titers become sufficiently high to bind 30% of an iodinated hCG standard. We report here the extensive characterization of the oLHbeta antibodies. Their binding affinity and capacity for human and rhesus gonadotropins were determined. The affinity was high (Ka - 10(9) M-1) and was similar for high, medium and low titer antisera. In contrast, the binding capacity for hCG correlated well with antibody titers (n = 18, r = 0.888, P less than 0.001) and ranged from 0.09 to 8.3 x 10(-7)M. Column chromatographic analysis showed that anti-oLHbeta was mostly IgG. A temporal increase of affinity ('maturation') after booster was absent in the majority of monkeys. Cross reaction between hCG and hLH as well as rhCG and rhLH was high. The latter, however, did not interfere with regular cycles and ovulation, suggesting that oLHbeta may be a useful antigen for human contraception. PMID- 6290657 TI - The validity of reporting results of cultures for herpes simplex virus after four days. PMID- 6290658 TI - Development of protective immunity against bacterial and viral infections in tumor-bearing mice coincident with suppression of tumor immunity. AB - This report addresses the question whether Meth A (methylcholanthrene-induced fibrosarcoma) tumor bearing Balb/c mice are able to develop specific antimicrobial immunity. Although specific suppressor T lymphocytes appeared during tumor growth which prevented expression of antitumor immunity, the development of protective immunity to L monocytogenes, S. pneumoniae or ectromelia virus infections was unimpaired. The Meth A tumor produced a soluble immunosuppressive factor which inhibited lymphocyte and macrophage functions in vitro. Tumor growth failed to inhibit the formation of immunoglobulin essential to antipneumococcal immunity, or the development of a specific acquired cellular resistance of primary importance in immunity to listeria and ectromelia virus infections. That tumor growth did not interfere with the development of cell mediated immunity was demonstrated by the effective transfer of antilisteria immunity by immune spleen from tumor-bearing mice. PMID- 6290659 TI - Lack of type C virus antibodies in systemic lupus erythematosus. AB - Twelve systemic lupus erythematosus (SLE) and 10 normal sera were similarly reactive in an assay for cytotoxic antibody to type C virus-infected and uninfected cells, but both were less cytotoxic for infected cells. Thus antibodies to type C viruses were not detected in SLE or normal sera, but type C infection may interfere with antibody recognition of other cell membrane antigens. PMID- 6290661 TI - Sodium periodate-induced human suppressor cells for polyclonal B cell activation. AB - Sodium periodate (SP) induces proliferation of mature T cells. In this study, human peripheral blood mononuclear cells (MNC) pretreated for 10 minutes at room temperature with increasing concentrations (0.1 to 5 mM) of SP before culture for 7 days in the presence of pokeweed mitogen (PWM) exhibited a dose-dependent inhibition of IgG production, contrasting with an increase in 3H-thymidine uptake. When MNC from 70 normal individuals were pretreated with 1 and 2 mM SP, IgG production in culture was suppressed by 46.8 +/- 4.5% and 60.4 +/- 4.4% (mean +/- S.E.M.), respectively, as compared to IgG synthesis in the presence of PWM alone. Longitudinal studies of MNC obtained from the same normal individuals over 6-10 months showed similar degrees of suppression, indicating that the level of SP-inducible suppressor cell activity remains relatively constant, although the degree of suppression varies among normal persons. Both T cells and monocytes were required for PWM-driven IgG production and for SP-induced suppression. A soluble factor elaborated by SP-treated monocytes was also able to suppress IgG production. This model should provide useful information about abnormal regulation of IgG synthesis in various pathological conditions. PMID- 6290660 TI - Thymic humoral factor effect on intracellular lymphocyte cAMP in patients with ankylosing spondylitis. AB - Intracellular cAMP was measured in peripheral blood mononuclear cells from patients with ankylosing spondylitis and from controls. The effect of thymic humoral factor (THF) on T lymphocyte cAMP content was monitored in both groups to determine if there were any differences in immature T cell proportions. Equivalent cAMP levels were found in patients and controls in the absence of THF and again after stimulation with this immune modulator. PMID- 6290662 TI - Susceptibility of squirrel and Cebus monkeys to foot-and-mouth disease virus. PMID- 6290663 TI - Repeated intravenous administrations of human albumin and human fibrinogen in the baboon: survival measurements. PMID- 6290665 TI - Clinical use of recombinant DNA techniques: the antenatal diagnosis of sickle cell anemia. PMID- 6290666 TI - Adenoid cystic carcinoma; a report of two cases. PMID- 6290664 TI - Enhanced sensitivity to stimulation of sodium transport and cyclic AMP by antidiuretic hormone after Ca2+ depletion of isolated frog skin epithelium. AB - The role of Ca2+ in the stimulation by antidiuretic hormone (ADH) of active sodium transport across the isolated epithelium of frog skin was investigated. This has been done by bathing the blood side with Ca2+-free solution containing 0.1 mM EGTA. This Ca2+ depletion halved the resistance but had no significant effect on the short-circuit current (SCC). The sensitivity of both cAMP- and SCC stimulation to ADH was increased 40-fold by Ca2+ depletion. Sensitivity to stimulation by theophylline was only changed a little, while stimulation by exogenous cAMP was completely unaltered. The increase in sensitivity to ADH was dependent on the duration of preincubation in Ca2+-free solution, which indicates that a slowly exchanging Ca2+ pool is involved in the determination of sensitivity to ADH. We suggest this pool is of cellular origin and the increased sensitivity is due to the decrease of a Ca2+ inhibition of the ADH-stimulated adenylate cyclase. But a direct effect of Ca2+ on binding of ADH to the receptor cannot be excluded. Our results are not compatible with the hypothesis that entry of extracellular Ca2+ is an obligatory step in the natriferic action of ADH, although it may be so in the hydroosmotic action of ADH. We also found the maximal response to ADH to be higher after Ca2+ depletion. This is in agreement with the hypothesis of intracellular Ca2+ as a modulator of the sodium permeability of the outward-facing membrane. PMID- 6290667 TI - Three-dimensional structures of cytochrome c oxidase vesicle crystals in negative stain. PMID- 6290668 TI - Relationship between membrane and cytoplasmic domains in cytochrome c oxidase by electron microscopy in media of different density. PMID- 6290669 TI - A systemic DNA sequencing strategy. PMID- 6290670 TI - Actin genes and actin messenger RNA in Acanthamoeba castellanii. Nucleotide sequence of the split actin gene I. PMID- 6290671 TI - Identification of polypeptides encoded by the replication of resistance factor R100. PMID- 6290672 TI - Testing models of the arrangement of DNA inside bacteriophage lambda by crosslinking the packaged DNA. PMID- 6290673 TI - Influence of cyclic nucleotides on the internal longitudinal resistance and contractures in the normal and hypoxic mammalian cardiac muscle. PMID- 6290674 TI - The cyclic AMP-dependent modulation of cardiac sarcolemmal slow calcium channels. PMID- 6290676 TI - The diffusion and electrogenic components of the membrane potential of rabbit ventricle after long exposure to different Ko. PMID- 6290675 TI - Antiadrenergic action of adenosine on ventricular myocardium in embryonic chick hearts. PMID- 6290677 TI - Glaucoma, hypertension, and marijuana. PMID- 6290678 TI - Accelerated appearance of chemically induced mammary carcinomas in obese yellow (Avy/A) (BALB/c X VY) F1 hybrid mice. AB - Latent periods and cumulative incidence of mammary carcinomas (MT) up to 50 wk after initial gavage with 7.12-dimethylbenz[a]anthracene (DMBA) were determined in virgin yellow (Avy/A) and agouti (A/a) (BALB/cStCrlfC3Hf/Nctr X VY/WffC3Hf/Nctr-Avy) F1 hybrid female mice. When subcutaneous masses reached 5-10 mm in diameter, the mice were killed and necropsied, and the tissues examined histologically. No MT were found in control mice. Cumulative MT incidence in the 1.5-mg DMBA group (A) was 43% (41/95) among yellow mice, and 33% (32/96) among agoutis. In the 6.0-mg DMBA group (B), corresponding MT incidences were 86% (83/96) and 71% (67/95). In group A, the first percentile of MT detection was 13.0 wk after initial carcinogen treatment in yellow mice; it was 18.0 wk in agoutis. Corresponding latent periods for the 20th percentile were 34.3 and 47.0 wk. In group B, latencies for the first percentile were 8.3 and 9.0 wk. Corresponding latencies for the 20th percentile were 15.3 and 16.0 wk. Within genotypes and dose groups, rates of weight gain of mice that developed MT and those that did not were similar. We conclude that MT induced by low doses of DMBA arise more rapidly in yellow mice than in nonyellow littermates. The absence of spontaneous MTs, acceleration of chemically induced MT formation at a low dose level that does not induce general toxicity, and availability of genetically identical (except for one gene) normal control animals make this experimental system suitable for development of an assay to efficiently test the carcinogenic potential of low dose levels of chemical substances. PMID- 6290679 TI - Metabolism of benzo[a]pyrene by fish cells in culture. AB - Benzo[a]pyrene (BaP) metabolism was studied in cell lines derived from rainbow trout (RTG-2), bluegill fry (BF-2), and fathead minnow (FHM). Confluent cultures were exposed to 3H-BaP (0.5 nmol/ml), and, after various exposure times, metabolites were extracted from the media with an organic solvent and analyzed by high-pressure liquid chromatography. BF-2 and RTG-2 cells converted 63% of the BaP to water-soluble metabolites within 24 h, while FHM cells converted only 12%. BF-2 and RTG-2 cells metabolized more than 90% of the BaP by 48 h, while only 67% of the BaP was converted to water-soluble metabolites by FHM cells after 96 h. The major organic-solvent-extractable metabolites in all three cell lines were 9,10-dihydroxy-9,10-dihydrobenzo[a]pyrene and unidentified polar metabolites. Of the water-soluble metabolites formed by BF-2, FHM, and RTG-2 cells, 67, 42, and 19%, respectively, were converted to ethyl-acetate-extractable metabolites by treatment with beta-glucuronidase. All three cell lines formed a glucuronide of 7,8-dihydroxy-7,8-dihydrobenzo[a]pyrene (7,8-diol); in BF-2 and FHM cells, the 7,8-diol represented almost half of the metabolites released by beta glucuronidase treatment. Thus, cell lines derived from three widely distributed species of freshwater fish have the capacity to metabolize BaP to a form that is a proximate carcinogen in rodents and to produce a water-soluble conjugate of this metabolite. PMID- 6290680 TI - Beta-adrenergic receptor binding in different regions of rat brain after various intensities of electroshock: relationship to postictal EEG. AB - Relationships among different intensities of electroshock treatment (EST), EEG recordable primary after discharge (PAD), and beta-adrenergic binding in various brain regions were investigated in adult rats. Different intensities of EST (1-2, 6-8, 20-30 mamp) were applied through ear-clip electrodes for 7 consecutive days. A sigmoidal relationship was observed between the PAD and the applied current intensity. 3H-dihydroalprenolol (DHA) was used as a ligand to evaluate beta adrenergic receptor binding in membrane preparation of the following brain regions: Cerebral cortex, cerebellum, pons-medulla, and "midregion"-which consisted of the remainder tissue including thalamus, striatum, and midbrain. A significant decrease (28%) in binding was observed in cortex tissue after EST of 6-8 mamp but no changes were observed after either 1-2 mamp or 20-30 mamp. In "midregion," a significant increase (40%) in binding was observed after EST with the subconvulsive intensity (1-2 mamp) whereas no changes were seen at higher intensities. The results suggest that in cortex, the function relating beta adrenergic binding to current intensities may not be monotonic. The increase in binding seen in the "midregion" after subconvulsive EST may not be associated with changes related to the clinical efficacy of EST. PMID- 6290681 TI - Synaptosomes and mitochondria from rat brain cerebral cortex: in vivo interference on some enzymatic activities by SAMe and CDP-choline. PMID- 6290682 TI - Determining the effective emetic dosage of tetrapotassium pyrophosphate (TKPP) in dogs. AB - The effect of TKPP on emesis was examined in adult "conditioned random source" mongrel dogs. Only dogs that exhibited emesis with CuSO4, were used. The effect of dosage and concentration of TKPP on emesis was highly significant. The effective combinations of dose and concentration were 4,800, 2,400, or 1,200 mg/head at 5, 10, or 20%. In all the tested dogs, 4,800 mg of 20% TKPP was effective inducing emesis. The mean latency of emesis in dogs was 7 min, 33 sec (95% confidence limits: 5 min, 58 sec 9 min, 7 sec). PMID- 6290683 TI - Adrenal response to repeated hemorrhage: implications for studies of trauma. AB - Endocrine responses to trauma are often inferred from responses seen after a single injury or elective surgery. However, major trauma frequently is followed by emergent surgery, necessitating evaluation of responses to repeated stimuli. We used sequential hemorrhage to study such responses. Splenectomized dogs anesthetized with pentobarbital (25 mg/kg) were hemorrhaged 7.5 ml/kg, 2 days following adrenal vein cannulation. Secretion rates of cortisol and catecholamines were determined in timed adrenal samples by HPLC. The bled volume was reinfused after 1 hour; the procedure was repeated 24 hours later. A significant response in adrenal secretion of cortisol was seen following hemorrhage each day (p less than 0.001), but the response on day 2 was 40% greater (p less than 0.05). Secretion rates of epinephrine and of norepinephrine did not change after hemorrhage on day 1 (p greater than 0.20). However, each hormone showed a dramatic response on day 2 increasing to 14x control levels (p less than 0.005). There were no differences in any cardiovascular variable during control period or after hemorrhage on the 2 days. The results demonstrate dramatic potentiation of the response to a second insult that persists for at least 24 hours after the first, and suggest that the endocrine response to traumatic injury with emergent surgery cannot be evaluated by studying responses either to isolated insults or to elective surgery. PMID- 6290684 TI - Exchange transfusion with Fluosol 43: in vivo assessment of cerebral cytochrome c oxidase redox state. AB - Rats were serially exchange transfused to hematocrits less than 2% (50-cc exchange) with either Krebs-Henseleit-albumin (KHA) solution or Fluosol-43 (F 43). Reflectance spectrophotometry was employed to monitor cerebral cytochrome c oxidase redox states. Determination of mean arterial pressure (MAP), arterial and venous oxygen content difference (A-VO2), and cerebral cortical ATP and lactate concentrations were also made. As hemodilution progressed, neither solution provided normal quantities of oxygen as evaluated by the cytochrome c oxidase redox state and a decrease in A-VO2 difference, although F-43 was significantly (p less than 0.01) better than KHA in terms of cytochrome c oxidase redox state as well as maintenance of MAP. There was no significant difference between the two test solutions in terms of cortical ATP and lactate. PMID- 6290686 TI - Other cerebellopontine angle (non-acoustic neuroma) tumors. PMID- 6290685 TI - Severe peripheral neuropathy after exposure to monosodium methyl arsonate. AB - A case of severe peripheral neuropathy after several days of exposure to a pesticide spray containing monosodium methyl arsonate (MSMA) is reported. The clinical manifestations of symmetrical peripheral neuropathy with stocking-glove sensory deficit, decreased position sense, decreased and absent deep tendon reflexes, and muscle wasting are consistent with those described in other cases of arsenic intoxication. The anemia, leukopenia, and bone marrow changes of dyserythropoiesis in this case are also similar to those previously described with arsenic intoxication. The authors discuss the possible contribution of toxicity from exposure to other pesticides. PMID- 6290687 TI - A novel structure seen when foot-and-mouth disease virus-induced poly (U) polymerase acts in a cell-free system. PMID- 6290688 TI - The control of freeze-drying with deuterium oxide (D2O). PMID- 6290689 TI - Bile duct adenocarcinoma in a pallid bat (Antrozeous pallidus). PMID- 6290690 TI - THC or analogues may alleviate some tremors. PMID- 6290691 TI - Unexplained fever in burn patients due to cytomegalovirus infection. PMID- 6290692 TI - Encephalomyocarditis virus myocarditis in inbred strains of mice--chronic stage. AB - Inbred strains of A/J, BALB/c, C3H/He, C57BL/6 and DBA/2 mice were inoculated with the M variant of encephalomyocarditis virus having a titer of 100 TCID50/0.1 ml. Myocardial lesions were seen in 73 of 150 BALB/c mice (48.7%), 160 of 259 C3H/He mice (61.8%) and 115 of 174 DBA/2 mice (66.1%). No pathologic findings were noted in A/J and C57BL/6 mice. In C3H/He and DBA/2 mice, dilatation and hypertrophy of the heart accompanying myocardial lesions persisted up to the 8th month after virus inoculation. The present study revealed that myocardial lesions similar to those in congestive (dilated) cardiomyopathy persisted for a long period after viral infection. PMID- 6290693 TI - Technetium-99m pyrophosphate uptake in spontaneously occurring perimyocardial lesions in an inbred strain of DBA/2 mice. AB - The spontaneous occurrence of perimyocardial calcification localized in the right ventricle has been reported in an inbred strain of DBA/2 mice. In this paper we examined the correlation between the myocardial uptake ratio of technetium-99m pyrophosphate (99mTc-PYP) and pathological findings in these mice ranging in age from one to 12 months and became lower with age, while calcification and fibrosis in the perimyocardium of the right ventricle became more prominent in aged mice. This may be due to an insidious onset of degeneration with calcification and fibrosis in spontaneously occurring perimyocardial lesions in these mice. Persistent abnormal uptake of 99mTc-PYP observed in the present study suggests the usefulness of such scintigrams in the diagnosis of chronic perimyocardial disease. PMID- 6290694 TI - [Clinical studies on obstetric analgesia by intrathecal beta-endorphin]. PMID- 6290695 TI - [Transfer of cefotiam into the tissues of gynecological organs]. AB - Uterine infections, particularly myometritis have been shown to be severe because of the difficult passage of the antibiotics into the affected tissues. Consequently, a biochemically novel drug, cefotiam (CTM) was investigated for the distribution into the uterine body, uterine cervix, ovary and oviduct of 10 patients with benign tumor, 1 g of CTM in 250 ml of 5% glucose was infused for 2 hours. The mean concentration of the antibiotics in the serum, uterine body, uterine cervix, ovary and oviduct at 30 minutes after the intravenous drip infusion was, respectively 19.7 micrograms/ml, 10.1 micrograms/g, 12.9 micrograms/g, 10.7 micrograms/g and 11.7 micrograms/g. The concentration ratio of the tissue to serum was found to be relatively high (51.5--65.5%), as compared to other antibiotics. In this experiments, no adverse effects, such as laboratory findings were found. PMID- 6290696 TI - [Concentration of cefotiam in the exudate of pericardium]. AB - The concentrations of cefotiam (CTM) in the exudate of pericardium were investigated, and determined in 8 patients accompanied open heart surgery after a single intravenous injection or intravenous drip infusion of 1 or 2 grams. The peak concentrations of CTM have been appeared after 30 minutes or 1 hour, and prolonged for 3 hours, these concentrations of CTM showed about 8--30 times of MIC values which inhibit the growth of E. coli, K. pneumoniae. P. mirabilis and S. aureus. Therefore, it is recommended that CTM will be administered by intravenous infusion just prior to the operation. PMID- 6290697 TI - [Clinical evaluation of cefotiam in internal medicine]. AB - Cefotiam (CTM) was administered to 52 patients with infectious disease associated with respiratory system, hematological malignancy, urinary system and other system. Good clinical responses were obtained in 38 out of 52 cases (73.1%). Neither objective and subjective side effects nor extreme abnormalities of laboratory tests were observed in these patients. It can be, therefore, concluded that CTM is 1 of the most useful drugs for infectious diseases in respiratory system, hematological malignancy, urinary system and other system. PMID- 6290698 TI - [Clinical evaluation of cefotiam chloride in the surgical field, with special reference to its intrathoracic distribution and prevention of postoperative infections following thoracic surgery]. AB - There are few report about the concentration of antibiotics in the tissue of intrathoracic organs. At present, it was analized the concentration in serum and tissue of intrathoracic organ and urinary excretion for CTM, and clinical effect against postoperative infection and side effect of CTM were discussed. 1) The peak concentration in serum was 48.7 micrograms/ml and half lives of CTM was about 30 minutes in 1 hour drip infusion of CTM 1g. 2) The concentration of CTM in lung tissue was about 30--50% of serum level. The concentration of CTM to pulmonary lesion was relatively high. 3) The concentration of CTM to the secretion of normal bronchiole was high as well. 4) The concentration of CTM to the bone marrow of rib was less than lung tissue. 5) Urinary excretion of CTM was 50--77% of total dose by 6 hours after injection. 6) CTM has wide spectrum and is useful to prophylaxis of postoperative infection. 7) It may be ineffective for Pseudomonas aeruginosa infection of lung. PMID- 6290699 TI - [Laboratory and clinical investigations of cefotiam in surgical biliary tract infections]. AB - Laboratory and clinical comparative investigations with cefotiam (CTM) and cefazolin (CEZ) were performed to confirm efficacy and safety in surgical, biliary tract infections. The following results were obtained. 1) The MICs of CTM against organisms, 20 strains, which were isolated from bile of patients with cholecystitis, were studied, especially those of CTM against E. coli and Klebsiella were 0.2 - greater than or equal to 100 micrograms/ml and 0.1--12.5 micrograms/ml, respectively, considerably lower than those of CEZ. And moreover against CEZ-resistant Proteus morganii and Serratia marcescens, CTM showed potent activities, that is, MIC values, 12.5--50 micrograms/ml and 0.78 microgram/ml, respectively. 2) In cholecystectomized patients, 2 grams of CTM was injected intravenously, followed by determination of bile concentration in gallbladder, common bile duct and concentration of gallbladder and liver tissue about 2 hours after administration. The mean bile concentrations of CTM in gallbladder and common bile duct were 1,213.2 micrograms/ml and 1,287.8 micrograms/ml, respectively. The peak concentration of CTM was 2,919.0 micrograms/ml. The mean concentrations of CTM in gallbladder and liver tissue were 28.5 micrograms/g and 45.7 micrograms/g, respectively. On the other hand, the mean bile concentrations of CEZ in gallbladder and common bile duct were 138.7 micrograms/ml and 128.8 micrograms/ml, respectively. 3) Bile concentrations of CTM was compared with those of CEZ by crossover method. The concentration of CTM was 37.7 micrograms/ml even at 5 or 6 hours after 2 grams intravenously administration. CTM showed extremely higher concentration than CEZ in bile. 4) The clinical effect was studied in 6 cases of surgical, biliary tract infections. The results were excellent in 2 cases, good in 3 cases and poor in 1 case, and the clinical efficacy was 83%. 5) CTM was administered to 6 patients who showed negative to intracutaneous reaction test. Nausea, itching, and eruption were observed in each 1 case after intravenously administration of CTM 2 g, however these adverse reactions disappeared within several hours. Throughout the course of treatment, any unusual laboratory findings related to CTM were not observed. PMID- 6290700 TI - [Therapeutic effects of cefotiam in biliary tract infections]. AB - As useful antibiotics for biliary tract infections, smooth transmigration to lesion and potent antibacterial activities will be demanded. Usually the antibiotics having potent antibacterial activities against E. coli and Klebsiella, which are considered to be main causative organisms of biliary tract infections, will be used for treatment. The concentrations of cefotiam (CTM) in bile and gallbladder wall tissue were investigated, and CTM showed very high concentrations in these tissues for several hours. And very good correlation between concentration of CTM in bile and laboratory findings, especially ICG R15 value, was obtained. CTM showed also very potent antibacterial activities against E. coli and Klebsiella. These results suggest that CTM will be useful agent for the treatment of biliary tract infections. PMID- 6290701 TI - [Clinical investigation of cefotiam against infections complicated by acute leukemia]. AB - Clinical investigation of cefotiam and aminoglycosides combination use against infections complicated by acute leukemia was performed, and the results obtained were as follows. 1) Sixteen patients were treated with cefotiam in doses 4--6 grams per day parenterally. The clinical effectiveness were excellent 47.6%, good 14.3%, fair 9.5% and poor 28.6%. No significant differences in therapeutic efficacy was observed between the patients given CTM 4 g/day and CTM 6 g/day. 2) It was considered that the clinical effectiveness of CTM has not been influenced by the difference of a number of maturation granulocyte. CTM has showed a certain, though not distinctive, efficacy compared with CFX. 3) As side effects with CTM, eruption appeared in 1 case, and no remarkable laboratory findings was observed. Cefotiam is thus considered to be a useful drug for the treatment of infections complicated by acute leukemia. PMID- 6290702 TI - [A clinical study on high dosages of cefotiam in serious infections associated with hematological disorders]. PMID- 6290703 TI - [Incompatibility of cefotiam dihydrochloride in parenteral preparation by high performance liquid chromatography]. AB - Change in external appearance, pH values and residual antimicrobial potency of cefotiam dihydrochloride (CTM) upon combination with infusion solutions and other injections was investigated. The combinations of CTM with bromhexine hydrochloride 2 mg/ml, 2 ml, dipyridamole 5 mg/ml, 2 ml and gabexate mesilate 100 mg/vial respectively were found to be incompatible on account of turbidity produced. As for 34 other combinations, no change in their appearance, pH and potency was observed. PMID- 6290704 TI - [Concentration of cefotiam in the cerebrospinal fluid]. AB - Cefotiam of 1 to 2 g was intravenously given during 15 to 60 minutes in 10 cases, and blood levels and cerebrospinal fluid (CSF) levels were studied. Following the drip infusion of cefotiam, maximum blood levels of 25.2 to 305 micrograms per ml was an average of 101.2 micrograms per ml were achieved at 15 to 60 minutes in 9 cases. Half life of cefotiam in serum was from 20 to 50 minutes, and mean time was 39.4 minutes. In contrast, maximum CSF levels of cefotiam were ranged 1.4 to 17.2 micrograms per ml and mean value was 5.7 micrograms per ml in 8 cases. The ratios of CSF levels to blood levels were calculated from 1.7 to 6.6% with an average of 4.6%. The CSF levels of cefotiam showed long delay and long decay. The period between the drip infusion and peak levels of cefotiam in CSF showed 60 to 420 minutes and mean time was 194 minutes. Half life varied between 45 and 270 minutes with an average of 178 minutes. No side effects were found in all cases. PMID- 6290705 TI - [Clinical bacteriological studies on cefotiam and cefsulodin in the field of otorhinolaryngology]. AB - Clinical bacteriological studies on cefotiam and cefsulodin in the field of otorhinolaryngology were carried out and the following results were obtained. 1) Aerobic and anaerobic Gram-positive bacteria were dominantly isolated from the clinical materials sent to the center from the clinical institutes. 2) It was considered that Streptococcus pneumoniae, Haemophilus influenzae and beta Streptococcus played an important role in the primary infections in the field of otorhinolaryngology. Staphylococcus aureus was also frequently isolated from the primary infections. Peptostreptococcus spp. was dominantly isolated from peritonsillar abscess. Gram-negative bacilli (GNB) were mainly isolated from the chronic secondary infections. Among GNB, Pseudomonas aeruginosa and Proteus spp. were more frequently isolated. Staphylococcus aureus was also constantly detected in the secondary infections together with GNB. Anaerobic bacteria were isolated from 20.1% of the patients with chronic otitis media and 27.1% of sinusitis. 3) Cefotiam showed potent antibacterial activities against most isolates of Gram positive and Gram-negative bacteria. 4) Cefsulodin showed potent antibacterial activities against clinically isolated Pseudomonas aeruginosa. Staphylococcus aureus and beta-Streptococcus were also susceptible to cefsulodin. PMID- 6290706 TI - [Clinical evaluation of tissue concentrations of cefotiam applied in the field of thoracic surgery]. AB - Seven patients who were performed thoracotomy for the disease of the chest, were administered cefotiam dihydrochloride 2.0 g for about an hour by intravenous drip infusion during their operation. Antibiotic levels of serum and intrathoracic tissues (a piece of lung parenchyma, parietal pleura, pulmonary hilar lymph node, chest wall muscle, pulmonary cyst and nerve) were determined, and an evaluation of bactericidal effect was discussed. In this study, we found that antibiotic level of poor blood supplied lung, so called the destroyed lung was remarkably high. This means that a high concentration of cefotiam dihydrochloride to intrathoracic tissue is effective against postoperative infection. PMID- 6290707 TI - [Clinical laboratory approach for determination of effective dose of cefotiam. Observation from disc susceptibility test and MICs]. AB - Reliability of cefotiam disc diffusion susceptibility test was investigated for determining whether the disc results correlate with minimal inhibitory concentration (MIC) or not. The results of Showa disc and Oxoid disc were well correlated. The inhibitory zone size of Showa disc and MIC were also well correlated, indicating reliability of the cefotiam disc test. The disc result, was observed over 84% of clinically isolated S. aureus, S. epidermidis, S. pneumoniae, S. pyogenes, E. coli, K. pneumoniae, P. mirabilis and H. influenzae. Their mean MICs were less than about 1.5 microgram/ml. Such a result should be useful for the determination of drug dose in various clinical conditions in order to obtain the proper drug levels in blood and tissues. The results of antibiotic disc diffusion susceptibility test with various bacteria isolated from clinical materials were compared. Cefotiam revealed a potent antimicrobial activity against Gram-positive cocci, showing a similar potency to that of cephalothin and cefazolin. However, it showed a stronger activity against Gram-negative bacilli than that of cephalothin, cefazolin, cephalexin and cefoxitin, revealing less resistant strains. Cephalosporins developed recently such as cefotaxime, cefoperazone, ceftizoxime, latamoxef etc. show very potent antimicrobial activity against Gram-negative bacilli, but their activity against S. aureus and S. epidermidis is weaker than that of cefotiam, cephalothin, cefazolin etc. Therefore, cefotiam can be considered as one of the drugs of first choice for surgical perioperative prophylaxis and blind therapy. PMID- 6290709 TI - [General pharmacological studies on cis-1-acetyl-4-[4-[[2-(2, 4-dichlorophenyl)-2 (1H-imidazol-1-ylmethyl)-1,3-dioxolan-4-yl] methoxy] phenyl] piperazine]. PMID- 6290710 TI - [Two cases of acute non-lymphoblastic leukemia treated with allogeneic bone marrow transplantation]. PMID- 6290711 TI - [Role of cyclic GMP in blood platelets]. PMID- 6290708 TI - [Experimental and clinical studies of cefmenoxime in the field of obstetrics and gynecology]. AB - The study group was organized to evaluate the usefulness of cefmenoxime (CMX) injection, a new synthetic cephalosporin, for the treatment of infections in the field of obstetrics and gynecology. Fundamental and clinical studies were made by the society and the following results were obtained. 1. The peak distribution of CMX's MIC for E. coli, Klebsiella sp., Enterobacter sp., Bacteroides sp. and Peptococcus sp. isolated from obstetrical and gynecological infections with relatively high frequencies area 0.1, less than or equal to 0.05, 0.2, 3.13, 1.56 micrograms/ml, respectively, with an inoculation of 10(6) cells/ml. 2. When 1 g of CMX is administered by intravenous drip infusion for 1 hour, the maximum concentrations in various tissues of female genital organs were as follows: 14.2 and 13.2 micrograms/g in ovary and oviduct, respectively, at 1.20 hours after the start of administration, and 16.9 and 26.3 micrograms/g in corpus uteri and cervix uteri, respectively, after 1 hour. As for the transfer to the exudate in the pelvic dead cavity, the peak concentration was 15.6 micrograms/ml after 2.13 hours. 3. In the clinical studies, CMX was given to 258 cases with female genital organ infections and others. As for the clinical effects, with exclusion of 3 cases in which other antibiotics are concomitantly used, responses were excellent in 76 cases, good in 162 cases and poor in 17 cases, among 255 cases in total. The efficacy rate was 93.3%. The efficacy rates by diseases were 97.1% (68/70) for intrauterine infections, 88.8% (79/89) for intrapelvic infections, 98.4% (62/63) for adnexitis, and 100% (23/23) for infections of external genital organs. As for the clinical effects on causative bacteria, the efficacy rates were 100% (19/19) for single infections due to Gram-positive bacteria, 94.8% (55/58) for single infections due to Gram-negative bacteria, and 88.2% (15/17) for single infections due to anaerobic bacteria. And its efficacy rates were 89.6% (69/77) for mixed infection cases. Side effects were observed in 2 cases (0.8%); 1 case with eruption, and 1 case with diarrhea and vomiting. As for abnormal laboratory findings, lower white blood cell count was observed in 2 cases and elevation of the values regarding hepatic functions in 9 cases. All cases were returned to the normal after the completion of the administration. Cefmenoxime showed a satisfactory clinical efficacy and a potent bacteriological effect in treatment of the infections in the field of obstetrics and gynecology, and it has been concluded that cefmenoxime will be useful addition to the antibiotics for the therapy of these infections. PMID- 6290712 TI - [Calcium activated neutral protease in the blood platelet]. PMID- 6290713 TI - [Platelet functions and cyclic nucleotide--studies with the use of inhibitors]. PMID- 6290715 TI - Immune response to alien histocompatibility antigens on Epstein-Barr virus transformed cells. AB - Expression of alien histocompatibility antigens on Epstein-Barr virus transformed, cultured lymphoblastoid cell lines (LCL), which were established from normal peripheral blood lymphocytes (PBL), was studied by means of mixed lymphocyte reaction (MLR), cell mediated lysis (CML) and primed lymphocyte typing (PLT). Stimulation of PBL by autologous LCL resulted in some MLR responses and generation of cytotoxic effector cells against autologous LCL. Restimulation of PBL by 16 individual allogeneic PBL failed to prime an individual lymphocytes against autologous LCL in PLT tests. However, stimulation of PBL by a pooled normal PBL resulted in generation of cytotoxic cells against autologous LCL. Culturing of stimulated PBL in the presence of T cell growth factor (TCGF) for 30 days was shown to maintain cytotoxic effector cells in the cell population. PMID- 6290714 TI - Enzymatic treatment of formalin-fixed and paraffin-embedded specimens for detection of antigens of herpes simplex, varicella-zoster and human cytomegaloviruses. AB - Attempts were made to detect antigens of herpes simplex (HS), varicella-zoster (VZ) and human cytomegalo (CM) viruses in pathological preparations made by formalin fixation and paraffin embedding as well as in infected culture cells by means of enzymatic treatment followed by indirect immunofluorescent staining, and the following results were obtained. 1) Specimens from autopsy cases, in which diagnosis was established by detection of viral antigens or virus isolation, showed specific antigens in specimens of various organs of all cases of HS (6 cases), VZ (3 cases) and CM (2 cases). In 17 other cases, which were suspected for those viral infections on the basis of ordinary light microscopic observation, also demonstrated respective viral antigens specifically, enabling differential diagnosis. In one case, HS antigen was detected in the esophagus and CM antigen in the liver, testifying occurrence of a double infection. 2) The time spun during which the specific antigens could be preserved was tested, and it was learned that preservation of stainable antigens was better in paraffin-embedded specimens than in specimens kept in formalin. It was possible to detect HS and VZ antigens in paraffin-embedded specimens prepared 20 and 18 years ago, respectively. 3) The optimal conditions of trypsin treatment varied depending upon the organ as well as upon each individual specimen, but a commonly recommendable procedure was found to by warming at 37 degrees C for 3 hours with 0.25% trypsin. A test with HS virus-infected Vero cells indicated that the effect of trypsin treatment was influenced by the concentration of formalin and the time of fixation. PMID- 6290716 TI - [Angiographic diagnosis of liver cancer]. PMID- 6290717 TI - [A study on carcinoma of the duodenal papilla: with special reference to the relationship to the relationship between radiological and pathological features]. PMID- 6290718 TI - [Clinicopathological studies on hepatic fibrosis in an area of high prevalence of hepatoma and liver cirrhosis]. PMID- 6290719 TI - [Clinical picture and geographical pathology of non-B type hepatocellular carcinoma]. PMID- 6290721 TI - [Experimental study on in vivo erythrocyte labeling with 99m Tc using stannous chloride]. PMID- 6290720 TI - [Local findings of renal cell carcinoma on 99m-Tc-DMSA renal, 99m-Tc-MDP bone and 67-Ga-citrate scintigraphies]. PMID- 6290723 TI - [Effects of the modes of sensory transmission on nociceptive responses--an experimental study based on the hypothesis by J.E. Johnson]. PMID- 6290722 TI - [Clinical significance of 99m Tc-HIDA scintigraphy in liver cell carcinoma and its metastatic foci]. PMID- 6290724 TI - Resting metabolism in hypoxia-acclimated rats during acute hypoxic exposure. AB - In contrast to the decreased metabolism in non-hypoxia-acclimated (CT) rats, acute hypoxia (9.5% O2) did not change the resting metabolism in hypoxia acclimated (HX: 12% O2 for 2 months) rats at 25 degrees C. After beta-adrenergic blockade, acute hypoxia greatly reduced the resting metabolism in HX rats. The calorigenic response to norepinephrine (0.4 mg . kg-1) during acute hypoxia was, however, identical with that observed in CT rats. These results suggest an enhanced beta-adrenergic stimulation in HX rats during acute-exposure to hypoxia. PMID- 6290725 TI - Reversible effects of monothiol (D-penicillamine) and dithiol (dimercaptosuccinic acid) chelating compounds on methylmercury-inhibited choline acetyltransferase activity and high affinity choline uptake. AB - The effects of thiol compounds on methylmercury chloride (MMC)-inhibited choline acetyltransferase (ChAT) activity and MMC-inhibited high affinity choline uptake of rat brain tissue were studied in vitro. D-penicillamine (D-Pc) and dimercaptosuccinic acid (DMS) reversed the MMC-inhibited ChAT activity dose dependently. Equilibrium dialysis of MMC-inhibited ChAT against the buffer containing 10(-3) M D-Pc reversed the ChAT activity almost completely. The reversal effect of D-Pc (monothiol compound) on MMC-inhibited ChAT was significantly more potent than that of DMS (dithiol compound). D-Pc and DMS almost equally reversed the MMC-inhibited high affinity choline uptake by synaptosomes in a dose dependent fashion. Washing with a solution containing D-Pc or DMS equally reversed the MMC-inhibited high affinity choline uptake in a dose dependent fashion. Neither D-Pc nor DMS could reverse the hemicholinium-3 inhibited high affinity choline uptake. PMID- 6290726 TI - Developmental changes in alpha-adrenergic and muscarinic receptor-mediated contractions of rat vas deferens. AB - Studies were made on developmental changes in phasic contractions of isolated rat vas deferens in response to norepinephrine (NE) and acetylcholine (ACh), which are mediated by alpha-adrenergic and muscarinic cholinergic receptors (alpha-R and m-R). The ED50 values of both contractions were almost constant during development, but the maximum responses to NE and ACh per wet weight and the relative contractions by NE and ACh to that by 100 mM K+ decreased greatly between 3 and 8 weeks after birth. However, the alpha- and m-receptors, assayed by measuring binding of [3H]-WB4101 and [3H]-quinuclidinyl benzylate (QNB), increased quantitatively per g wet weight and per mg protein between 2 and 8 weeks after birth, but did not change qualitatively. Thus, a discrepancy between the increases of alpha- and m-receptors and decreases of the maximum responses to NE and ACh was found, and possible mechanisms for this are discussed. PMID- 6290727 TI - Operative results of extrahepatic bile duct carcinoma. AB - To determine the benefits of surgical treatment for patients with carcinoma of the extrahepatic bile duct, data on 100 patients with this disease who had been surgically treated in our clinic during the past 18 years were evaluated. These patients were grouped into three, i.e., upper, middle and lower bile duct groups. Patients with periampullary tumor were excluded from this study. Surgical procedures consisted of resection of the tumor, including hepatic resection and dissection of the regional lymph nodes, and a bypass operation of the extra- or intrahepatic bile duct. Resectability rates of the tumor were 21.6% in upper, 82.4% in middle, and 50% in lower bile duct groups. Average survival times of patients who had a resection of the tumor were 30.3 months in the upper bile duct group, 35.9 months in middle (the longest, 13 years and 3 months) and 22.5 months in lower bile duct group. Survival rates of overall patients with resection of tumors were 64.5% at one year, 29.0% at 3 years, and 12.9% at 5 years after surgery, respectively. The middle bile duct group showed the most favorable operative results of all the groups, an extended resection of the tumor should be carried out to obtain for a longer survival. PMID- 6290728 TI - Relationship between the content of estrogen and progesterone receptors and the pathological characteristics in human breast cancer. AB - Assays of estrogen receptors (ER) and progesterone receptors (PgR) were performed by using the dextran-coated charcoal (DCC) method in 124 cases of invasive breast cancer. The results were correlated with clinical and pathological characteristics. There was no correlation between steroid hormone receptor contents and menopausal status, size of tumor, axillary lymph node status, or histological type. The presences of ER and PgR were significantly correlated with histological grade and its mitotic component. 78.3% of well-differentiated (Grade I) tumors were ER positive. Of this number, 61.1% were also PgR positive. In contrast, 69.0% of poorly differentiated (Grade III) tumors were ER and PgR negative. Tumors with a prominent lymphoid infiltration demonstrated a low frequency of positive ER and PgR. There was a significant inverse correlation between the degree of lymphoid infiltration and histological grade. These results suggest that the ER and PgR status of tumors may indicate a malignancy, and prognostic information can thus be obtained independently of other known factors such as size of the tumor and axillary lymph node status. PMID- 6290729 TI - [Clinical significance of serum levels of angiotensin-converting enzyme and lysozyme in sarcoidosis, with special reference to its prognosis and reaction to steroids]. PMID- 6290730 TI - [A case of resected solitary pulmonary metastasis and local recurrence 10 years after the removal of submandibular adenoid cystic carcinoma]. PMID- 6290731 TI - Myristate-induced release of superoxide and hydrogen peroxide from peritoneal macrophages in mice immunized to Toxoplasma gondii and Plasmodium berghei. PMID- 6290732 TI - Equine Getah virus infection: pathological study of horses experimentally infected with the MI-110 strain. PMID- 6290733 TI - A pathologic study on fetuses and placentas from cows affected with enzootic bovine leukosis with reference to transplacental infection of bovine leukemia virus. PMID- 6290734 TI - Expression of fibronectin and laminin in the rat liver after partial hepatectomy, during carcinogenesis, and in transplantable hepatocellular carcinomas. AB - The distribution in the F344 rat liver of two extracellular matrix and basement membrane components, fibronectin and laminin, was studied by immunofluorescence. Fibronectin was found diffusely in normal liver lining the sinusoids and in connective tissue surrounding blood vessels and bile ducts; laminin was present predominantly in the basement membranes of blood vessels and bile ducts and was only inconsistently seen lining the sinusoids. After partial hepatectomy (PH), there was a transient decrease of fibronectin in the central and midzone sinusoidal hepatic areas. This decrease was most marked on day 3 after the PH. Carcinogens caused marked changes in the distribution of fibronectin. Large extracellular deposits of fibronectin were seen in areas of oval cell proliferation in livers of rats treated with N-2-fluorenylacetamide (2-FAA) while being fed a choline-deficient diet. In contrast, the nodules that developed in these livers were almost completely devoid of fibronectin staining. Neoplastic nodules produced in rats by cyclic feedings of 2-FAA r by injections of diethylnitrosamine also contained little or no fibronectin. Laminin staining did not change markedly during these treatments, but increased staining was seen associated with the newly formed ductlike structures and oval cells in liver of rats treated with carcinogens. Transplantable hepatomas varied in their fibronectin staining from fibronectin-negative hepatomas to ones with fibronectin staining within or around every tumor cell. Laminin was only found around the vascular structures within the tumors. The presence or absence of fibronectin in hepatomas did not show an obvious correlation to growth rate or metastatic potential of the tumors studied. PMID- 6290735 TI - Spontaneous leukemia viruses: lymphomagenic ecotropic viruses of AKR mice. AB - The spontaneous leukemia (SL) viruses are ecotropic lymphomagenic viruses isolated from AKR spontaneous lymphomas. These viruses are produced stably by continuous cell lines from spontaneous lymphomas and by a cell line derived from the bone marrow stroma of an AKR mouse neonatally inoculated with an SL virus. All cell lines cloned from the parent lymphoma cell lines consistently produce SL viruses. These viruses can be passaged in vivo and maintain their leukemogenic properties. Cloned isolates of SL viruses accelerate lymphoma in AKR mice and induce thymic lymphoma in mice of other strains. Thus their lymphomagenic properties are conclusively shown. In a study with the use of a sensitive host range assay, xenotropic and/or dual-host range viruses are consistently found in spontaneous lymphoma and cell lines derived from them. However, viruses able to replicate in mink lung cells are not expressed in SL virus-induced lymphomas or their derived cell lines. PMID- 6290736 TI - [The menopause]. PMID- 6290737 TI - The effect of electroconvulsive therapy on plasma cyclic-AMP, non-esterified fatty acid, tryptophan and tyrosine in depression. PMID- 6290738 TI - [Usefulness of scintigraphy with the use of Tc-99m-labeled pyrophosphate in the diagnosis of a fresh myocardial infarction]. PMID- 6290739 TI - [Diagnosis of acute myocardial infarction by means of scintigraphy with technetium 99m-labeled pyrophosphate]. PMID- 6290740 TI - [Diagnosis of complications arising after cardioversion and reanimation by means of myocardial scintigraphy using technetium 99m-labeled pyrophosphate]. PMID- 6290742 TI - In vivo effect of cellular calcium uptake on osmotic and nonosmotic release of arginine vasopressin. PMID- 6290743 TI - Choline kinase activity along the rabbit nephron. AB - Choline kinase catalyzes the phosphorylation of choline to phosphorylcholine which is thus made available for phosphatidylcholine biosynthesis. Choline kinase activity was determined in defined microdissected structures of rabbit nephron with a radiochemical microprocedure. Enzyme activity was present in all segments tested. When referred to tubular length, the highest activities were found in proximal convoluted tubules. Due to the lower protein content of distal structures these segments exhibited higher enzyme activities when referred to micrograms of protein. From this distribution pattern it was concluded that all nephron segments are able to use extracellular choline for phosphatidylcholine biosynthesis. PMID- 6290744 TI - Sodium homeostasis in chronic renal disease. PMID- 6290741 TI - Elevated urinary PCO2 in the rat: an intrarenal event. AB - During a bicarbonate diuresis, final urine Pco2 considerably exceeds systemic Pco2, an effect thought to reflect the postpapillary delayed dehydration of carbonic acid. To test this explanation, Pco2 tensions along the inner medullary collecting duct (IMCD) of bicarbonate-loaded rats were measured directly using Pco2 microelectrodes. With systemic Pco2 held at 40 mm Hg, IMCD Pco2 exceeded systemic Pco2 in every measurement by an average of 20 to 30 mm Hg. A significant increment in Pco2 was seen between 50% IMCD length and the papilla tip. During the infusion of carbonic anhydrase, IMCD Pco2 was reduced but not to systemic levels. Finding elevated Pco2 along the terminal IMCD deemphasizes the importance of postpapillary delayed dehydration and suggests the possibility that bicarbonaturia is associated with papillary accumulation of carbon dioxide. PMID- 6290745 TI - Suppressor cells assayed by numerical and functional tests in chronic renal failure. AB - Suppressor cells were assayed by numerical and functional tests in adults on chronic hemodialysis. Peripheral blood mononuclears (PBM) were classified as total T-cells by E-rosettes and by the monoclonal antibody OKT3, as T-cell subsets by OKT4 (inducer/helper T-cells) and OKT8 (cytotoxic/suppressor T-cells) and as B-cells by the presence of surface immunoglobulin. The suppressive effect of PBM pretreated with either Concanavalin A (Con A), sodium periodate, or serum rich in immune complexes, on normal homologous phytohemagglutinin (PHA) lymphocyte transformation, was determined. Usual tests of T-cell function were not done. T lymphopenia was due to significant diminution (P less than 0.002) in numbers of OKT4+ cells in patients (516 +/- 44 cells/mm3, mean +/- sem) as compared to controls (906 +/- 96 cells/mm3). The number of OKT8+ cells in patients was not different from normal although their percentage (45 +/- 4%) was slightly higher than controls (36 +/- 5%) (P less than 0.10). Suppressor activity using only a suboptimal dose of Con A (5 micrograms/ml), was significantly lower (P less than 0.002) in uremic patients (36 +/- 12%) than in controls (67 +/- 7%). An important finding was that no significant correlations were detected between the numerical and functional assays of suppression used or between any of these immunological tests and biochemical parameters studied. The implications of these results for immunoparesis in uremia are discussed with particular reference to the discordance between marker and functional assays of suppressor cells. PMID- 6290747 TI - Histochemistry of angiotensinase A in the glomerulus and the juxtaglomerular apparatus. AB - The aminopeptidase A (APA; angiotensinase A, E.C. 3.4.11.7) can be demonstrated histochemically (by simultaneous azo coupling) in the glomerulus and JGA of the rat and mouse kidney by light and electron microscopy. In the rat JGA, the APA is localized in Goormaghtigh's cells (mainly at the cell membranes), and in the mouse it is to be found in epitheloid cells (at the cell membranes and in lysosomal structures). From quantitative histochemical investigations of subcortical glomeruli, it can be proven that the aminopeptidase A determined by histochemical means is the angiotensinase A that splits N-terminal aspartate from AI or AII. Animal experiments (sodium depletion, sodium loading, adrenalectomy) bring about marked APA alterations in the glomeruli and in the JGA. The strongest APA alterations can be seen in the adrenalectomized animals, namely an increase of the APA activity in the glomeruli and parts of the JGA. The findings suggest that this enzyme is important for the regulation of the glomerular blood flow and the renin secretion or production by means of angiotensin degradation. PMID- 6290746 TI - Immunocytochemistry of the renin-angiotensin system: renin, angiotensinogen, angiotensin I, angiotensin II, and converting enzyme in the kidneys of mice, rats, and tree shrews. PMID- 6290748 TI - [Use of iruxol in surgical clinical practice]. PMID- 6290749 TI - [Influence of local application of isoptomax eye drops on neuromuscular transmission]. AB - The muscle-relaxing effect of antibiotics, particularly that of aminoglycosides and polymyxins after oral, intravenous, intramuscular and local application is described. After local application of Isoptomax eye drops, one patient suffered from relaxation of the muscular system of the neck and throat. The phenomenon did not recede until several hours after therapy was discontinued. The mechanism of action is described and a possible treatment discussed with reference to the relevant literature. PMID- 6290750 TI - [Virological diagnosis of ocular adenovirus infection]. AB - For laboratory diagnosis, virus isolation and identification from conjunctival swabs should be combined with serology (two blood specimens: early and 14 days or later after onset of disease). The group-specific complement fixation fails in almost one-half of the cases. Additional type-specific reactions (neutralization, hemagglutination inhibition, type-specific complement fixation) may be performed with adenovirus 8 and, in epidemic outbreaks, also with other types. Immunofluorescence procedures have proved to be of limited value; rapid diagnostic procedures are not yet available. PMID- 6290751 TI - Problems caused by pharmacologic glucocorticoid treatment of nephrotic syndrome. PMID- 6290752 TI - [Pathology of soft tissue sarcomas: 238 cases of the childhood tumor registry]. AB - Until April 1981 malignant soft tissue sarcomas were registered from 238 patients. Rhabdomyosarcoma was the most common tumor (115/238 = 48.3%). The embryonal subtype was predominantly seen among the rhabdomyosarcomas (83/115 = 72.2%). Rhabdomyosarcomas were localized most frequently in the head and neck area (40/115 = 34.8%), followed by genitourinary system (15/115), pelvis soft tissue (12), abdomen (10) and extremities (10). Non-rhabdomyosarcomatous soft tissue sarcomas (123/238 = 51.7%) were synovial sarcomas (20 = 8.4%), fibrosarcomas including spindle cell sarcoma (17 = 7.4%), leiomyosarcomas (12 = 5.0%), malignant tumors of the vascular system (11 = 4.6%) and neurofibrosarcomas (9 = 3.8%). Other types of sarcoma were extremely rare. 42 (17.6%) of all soft tissue sarcomas could not be classified histogenetically. Rhabdomyosarcomas could be diagnosed much more accurately (105/115 = 91.3%), compared to all other soft tissue sarcomas (99/121 = 81.8%). At present, the most difficult diagnostic problems remain with the tumors of connective tissue, in particular with fibrosarcomas and with the differential diagnosis of juvenile fibrosarcomas versus juvenile fibromatoses. PMID- 6290755 TI - [Trends in the development of clinical endocrinology in the eighties]. PMID- 6290753 TI - [Islet cell cancer with organic hyperinsulinism. Clinical aspects, diagnosis and therapy]. AB - About 8%-15% of the patients with organic hyperinsulinism have an islet cell carcinoma (13% in our series). In addition to a history of complaints of relatively recent onset, the patients present clinically the typical intermittent neurologic-psychiatric symptoms concurrently associated with hypoglycemia. The diagnosis is established biochemically on the basis of hypoglycemia, with inadequate incrementation of the insulin concentration subsequent to suppression and provocation tests. Elevated serum proinsulin and, in most patients, an increased insulin secretion rate are usually found after administration of agents such as glucose or leucine. Localization of the tumors is achieved by selective coeliacography as well as abdominal computerized axial tomography. The islet cell carcinoma is found most frequently in the tail of the pancreas, less frequently in the body and head of the pancreas. Metastatic spread is seen early into adjacent lymph nodes and especially in the liver. The treatment of choice is surgical resection of the tumor. Even in cases with advanced metastatic involvement, surgical intervention appears indicated. Medical treatment includes the administration of diazoxide, long-acting glucagon as well as the cytostatic agent streptozotocin. The average survival time is 30-40 months after diagnosis (in our series 79 months). Thus, the prognosis of patients with islet cell carcinoma appears relatively favorable, especially when compared with adenocarcinoma of the pancreas. PMID- 6290756 TI - [Maternal happiness has many question marks]. PMID- 6290758 TI - ["Krankenpflege" offers for discussion: part-time work]. PMID- 6290759 TI - [Part-time work in the hospital: attempt at an objective opinion]. PMID- 6290757 TI - [After care and rehabilitation: challenge to medicine and nursing]. PMID- 6290754 TI - [Short-term induction and cyclic maintenance therapy in inoperable small cell bronchial cancer]. AB - Since July 1978 one hundred and three consecutive patients with unresectable small cell bronchogenic carcinoma were treated with a combination of doxorubicin, cyclophosphamide and vincristine (ACO). In limited disease patients (64) the second chemotherapy course was followed by prophylactic cranial irradiation, the fourth by irradiation towards primary disease sites. Complete responders were randomised to either receive etoposide or no further maintenance therapy. Objective responses were reached in 88/100 evaluable patients, with 72% of complete remissions in limited-stage disease and 33% in extensive disease, respectively. The actuarial median survival time for limited disease patients was 15.8 months compared to 9.3 months in extensive disease (p less than 0.005). 29 of the 100 patients remain still alive, 4 for more than 24 months without disease recurrence. The survival advantage of patients reaching complete remissions relative to those who did not is highly significant (p less than 0.001). Acute gastrointestinal and hematological side effects were common, with possibly three drug-related deaths from infections during transient granulocytopenia (mean nadir: 600-900 cells/mm3). The present induction regimen using only four courses of chemotherapy produces high complete remission rates on roentgenography and bronchoscopy and improved survival in the majority of patients. Thus far any effectiveness of etoposide-maintenance therapy following ACO could not be substantiated. PMID- 6290760 TI - [Part-time work. Which motives justify it?]. PMID- 6290761 TI - [4 nurses come before the court. "Equal pay for equal work"]. PMID- 6290762 TI - [Patients' rights. The dignity of man]. PMID- 6290763 TI - [Patient's right]. PMID- 6290764 TI - [Patients' rights. Information]. PMID- 6290765 TI - [Rights of the hospitalized child]. PMID- 6290766 TI - [Patients' rights. The hospital is not a 3-star hotel]. PMID- 6290767 TI - [Patients' rights. To be back in charge]. PMID- 6290768 TI - [Patients' rights. The voice of health care consumers]. PMID- 6290769 TI - [Patients' rights. The right to economic care]. PMID- 6290770 TI - [At the nursing school of the Vaud University Hospital Center. A much celebrated 50th anniversary]. PMID- 6290771 TI - [The effect of words; their representation and utilization by a professional corps in search of themselves]. PMID- 6290772 TI - [19th century Ticino and the cholera]. PMID- 6290774 TI - [Violence. Submission to authority]. PMID- 6290773 TI - [Amnesty International: first international meeting of AI nurses]. PMID- 6290775 TI - [Public health nurses of Vaud and Fribourg cantons in their patient contacts]. PMID- 6290776 TI - [Visiting nurse, a new profession accredited with a diploma]. PMID- 6290778 TI - [Educational structure. Models of education. Initial results]. PMID- 6290777 TI - [A nursing future in home care services]. PMID- 6290779 TI - [Patients' rights. The Swiss Nurses' Association/Geneva participates in the launching of the proposal for the rights of patients]. PMID- 6290780 TI - [The Swiss Air Rescue Guard. Corporate member of the Swiss Red Cross]. PMID- 6290781 TI - [American nurse, are you better off?]. PMID- 6290782 TI - [Why do students who love nursing become resigned nurses?]. PMID- 6290783 TI - [Operating room nursing. Human weaknesses and incidental human weaknesses in the operating room]. PMID- 6290784 TI - Biology of disease: free radicals and tissue injury. AB - A free radical is any molecule that has an odd number of electrons. Free radicals, which can occur in both organic (i.e., quinones) and inorganic molecules (i.e., O(2)), are highly reactive and, therefore, transient. Free radicals are generated in vivo as by products of normal metabolism. They are also produced when an organism is exposed to ionizing radiation, to drugs capable of redox cycling, or to xenobiotics that can form free radical metabolites in situ. Cellular targets at risk from free radical damage depend on the nature of the radical and its site of generation. In this review we survey cellular sources of free radicals and the reactions they can undergo and discuss cellular defenses and adaptive mechanisms. PMID- 6290785 TI - Human papillomavirus. Frequency and distribution in plantar and common warts. AB - Fifty-nine (24 plantar, 35 common) warts surgically excised from 44 patients (15 female, 29 male), 18 to 32 years old, were examined by electron microscopy (EM) for papillomavirus particles and by fluorescent antibody (FA) and peroxidase antiperoxidase (PAP) tests for human papillomavirus type 1 (HPV-1)-specific antigens and papillomavirus genus-specific (common) antigens. Fifty per cent of plantar warts and 52 per cent of common warts were positive for HPV by EM. When examined by FA and PAP for genus-specific antigens, 58 per cent of plantar warts and 68 per cent of common warts were positive. Fifty per cent of plantar warts and 11 per cent of common warts were caused by HPV-1 as determined by reactivity with HPV-1 type-specific antiserum. All warts positive by EM were positive by FA and PAP. Sampling error accounted for warts positive by FA and PAP but not by EM. Forty-two per cent of plantar warts and 31 per cent of common warts were negative by both EM and FA and PAP. The warts caused by HPV-1 contained more virus and were more clinically aggressive than warts caused by other HPV. This study shows that the results obtained by the PAP technique using papillomavirus genus- and type-specific antisera on formalin-fixed, paraffin-embedded tissue available to the diagnostic pathologist are concordant with the results obtained by methodology usually available only to the experimental pathologist. PMID- 6290786 TI - Effect of hormone injections on levels of cytosolic receptors for estrogen, androgen and progesterone in dog prostate. AB - We have studied the effects of estradiol injections on cytosolic estrogen, androgen and progesterone receptor levels in order to understand the role of this steroid in the induction of prostatic hyperplasia in the dog. Adult mongrel dogs were castrated and were then given injections of estradiol (0.25, 0.8 or 2.5 mg) dissolved in olive oil containing 5% benzyl alcohol on days 0, 2, 5 and 7 after castration. Steroid receptor levels were determined by Scatchard analysis using charcoal assay one day after the last injection. All three receptors were increased maximally with the 0.8 mg dosage when compared with castrated controls. Estradiol binding increased from 89 +/- 10 (mean +/- SEM) to 361 +/- 37 fmol per mg prot., androgen binding from 47 +/- 5 to 123 +/- 11 fmol per mg prot. and progesterone binding from 26 +/- 3 to 211 +/- 36 fmol per mg prot. The small dose of estradiol. (0.25 mg) produced a significant (P less than 0.05) increase of the progesterone receptor levels from 25 +/- 3 to 48 +/- 14 fmol/mg prot. Substitution of estradiol by 5 alpha-androstan-3 beta,17 beta-diol (2.5 or 25 mg) resulted in receptor levels similar to castrated animals. However treatment with 5 alpha-androstan-3 alpha,17 beta-diol (25 mg) alone or in combination with estradiol (0.25 mg) increased significantly the androgen receptor while it decreased the estrogen receptor. These results show that the administration of estradiol at doses used to induce experimental prostate hyperplasia produce measurable effects in the prostate and suggest that the estradiol receptor may be implicated in this phenomenon. PMID- 6290787 TI - Androgen and progestogen production in cultured interstitial cells derived from immature rat testis. AB - Interstitial cells derived from intact immature rats were cultured as monolayers. Their response to gonadotropins was evaluated by radioimmunoassay of 3',5'-cyclic AMP and steroids in the medium. Steroids were measured either directly (testosterone and progesterone) or after previous oxidation and thin layer chromatographic purification of the steroid extracts (4-androstene-3,17-dione, 5 alpha-androstane-3,17-dione, progesterone, 5 alpha-pregnane-3,20-dione). It could be demonstrated that these cells respond to gonadotropins with increased secretion of C19- and C21-steroids for at least 10 days. The total amount of steroids secreted in the medium, however, decreases markedly. During the first days of culture C19-steroid production falls dramatically whereas the secretion of C21 derivatives increases. A major fraction of the extracted steroids has undergone 5 alpha-reduction. A characteristic feature of cultured interstitial cells is the bell-shaped profile of the dose-response curve for gonadotropin stimulated androgen production. This profile is the result of a steroidogenic lesion situated at the level of the 17 alpha-hydroxylase and/or 17,20-desmolase and induced by high concentrations of gonadotropins. Daily changes with medium supplemented with LH or FSH, initiated on day 3 of culture, prevent a further loss of steroidogenic potential, restore the ability to produce C19-derivatives, and tend to normalize the dose-response curve for gonadotropin stimulated production of androgens. PMID- 6290788 TI - In vitro metabolism of testosterone to 17 beta-hydroxy-5 alpha-androstane-3-one and 5 alpha-androstane-3 alpha, 17 beta-diol by the rat intestine. AB - The metabolism of testosterone to 17 beta-hydroxy-5 alpha-androstane-3-one and 5 alpha-androstane-3 alpha, 17 beta-diol by the 800 g supernatant fraction by different parts of the gastrointestinal tract from male rats was investigated. This metabolism tended to be higher in immature than in mature animals. Administration of dexamethasone or long-acting ACTH to immature and mature rats increased testosterone metabolism to 17 beta-hydroxy-5 alpha-androstane-3-one and 5 alpha-androstane-3 alpha, 17 beta-diol by ileum tissue. No such effect could be observed following administration of progesterone, estradiol, prolactin, LH or FSH in mature animals. Development of the gastrointestinal tract from the immature to the mature stage was associated with augmented metabolism of testosterone to 17 beta-hydroxy-5 alpha-androstane-3-one and 5 alpha-androstane-3 alpha, 17 beta-diol in the ileum. PMID- 6290789 TI - Evaluation of tumor aggressiveness in breast carcinoma according to a scoring method. AB - A scoring system was used to classify 310 patients with breast carcinoma. Tumors were divided and scored according to their architectural patterns and histological features of known prognostic significance such as tumor volume, nuclear grade, mitosis, etc. According to the scoring value, tumors were classified as aggressive (A), moderately aggressive (M), and low aggressive (L). Results showed a higher rate of metastasis in patients with A tumors (72%) and M tumors (63%) when compared with L tumors (22%). There were 175 patients with axillary lymph node metastasis. Of these, 101 had A tumors, 91 had recurrences (90%), and 81 died (80%); of the 56 patients with M tumors, 26 had recurrences (46%) and 9 died (16%) (P less than 0.001). None of the 18 cases classified as L recurred (P less than 0.001). There were 135 patients with no nodal metastasis: 39 had A tumors (21 recurrences [54%] and 19 deaths [49%] and 96 had M and L tumors (1 recurrence [1%] and no deaths) (P less than 0.001). PMID- 6290791 TI - Patterns of breast cancer detection in the United States. AB - The American College of Surgeons Commission on Cancer Short-term Survey of Breast Cancer in 12,315 patients showed that 73% of malignant tumors are found by patients, 23% by physicians, and 4% by mammography. It also indicated that younger women are more likely to discover tumors than older women, and that mammography is more likely to detect small tumors with negative axillary nodes. The effectiveness of mammography is most evident in women 50 to 74 years of age, although in women 45 to 49 years, the frequency of tumors detected by mammography nearly equals that for other groups. In black women, mammography may not be currently fully utilized. Analysis of the survey data would indicate that patients appear to demonstrate adequate skill in detecting tumors, as compared to physicians. A delay in diagnosis of longer than 3 months is associated with larger tumors and increased likelihood of axillary metastasis. PMID- 6290790 TI - Long-term intraarterial chemotherapy infusion of ambulatory head and neck cancer patients. AB - The complications of percutaneous intraarterial infusion chemotherapy of the head and-neck-cancer patient has dampened enthusiasm for this approach. A totally implantable infusion pump system circumvents many of these complications and will in the long term enhance the opportunity to expand upon the advantages of infusion chemotherapy for the benefit of all cancer patients. PMID- 6290792 TI - [Interaction between calcium antagonists and vascular postsynaptic alpha receptors]. AB - 1. The present review deals with the vascular action of calcium antagonists (CA) at the cellular level, emphasizing the interference between CA and postsynaptic alpha 2-adrenoceptors in the resistance vessels. 2. This interference, which can be demonstrated in various animal species, implicates that the pressor effect of an alpha 2-adrenoceptor agonist is diminished by the CA and by N alpha 2-EDTA. However, the vasoconstriction initiated by alpha 1-adrenoceptor agonists is hardly reduced by CA. Accordingly, the interaction is selective for alpha 2 adrenoceptors. 3. The following hypothesis is submitted: After formation of the alpha 2-adrenoceptor/agonist complex there occurs a transmembranous influx of extracellular calcium ions. This influx, which is inhibited by CA, is probably indispensable for the activation of contractile proteins. 4. Accordingly, the vasodilator activity of the CA might be explained as follows: the CA inhibit the alpha 2-receptor mediated component of the vasoconstriction, induced by endogenous catecholamines, without influencing the alpha 1-receptor component of the pressor effect. PMID- 6290793 TI - [Comparative study of piperoxan and 2-(2-imidazolinyl)-1, 4-benzodioxane (170 150) on pre- and postsynaptic receptors in the rat]. AB - 1. 170 150 (imidazolinyl-2)-2-benzodioxane 1-4), as does piperoxan, competitively antagonizes the hypertension induced by clonidine in the pithed rat. Piperoxan appears slightly less potent than 170 150 in this preparation as shown by the comparison of the apparent pA10 values: 5.3 for piperoxan versus 5.4 for 170 150. 2. The two drugs antagonize the reduction of the electrically-induced tachycardia produced by clonidine. 170 150 appears to be 3-fold more potent than piperoxan in this preparation. 3. These results are compatible with a blockade of alpha 2-pre and postsynaptic adrenoceptors of the rat by piperoxan and 170 150 appears to be 3-fold more potent than piperoxan in this preparation. 3. These results are compatible with a blockade of alpha 2-pre and postsynaptic adrenoceptors of the rat by piperoxan and 170 150 and they are in agreement with our previous results which indicate that compound 170 150 shows a preferential affinity for alpha 2 adrenoceptors. PMID- 6290795 TI - Theory of metabolic regulation including hormonal effects on the molecular level. PMID- 6290796 TI - A proton motive force transducer and its role in proton pumps, proton engines, tobacco mosaic virus assembly and hemoglobin allosterism. PMID- 6290794 TI - [Modulator effect of N6-monobutyryl-cAMP in experimental inflammation of rat conjunctiva]. PMID- 6290797 TI - Electronic aspects of enzyme catalysis proton-electron density displacements. PMID- 6290798 TI - Muscimol and some aza analogues: molecular orbital calculations relating to their potency as GABA mimetics. PMID- 6290799 TI - DNA cruciform structures: implications for telomer replication in eukaryotes and instability of long palindromic DNA sequences in prokaryotes. PMID- 6290800 TI - A model for the mechanism and control of eukaryote gene splicing. PMID- 6290801 TI - Generation of models of protein structure from solution scattering experiments. PMID- 6290802 TI - Influence of histologic type on survival after curative resection for undifferentiated lung cancer. AB - Undifferentiated carcinoma of the lung carries a worse prognosis overall than other cell type, but it is unclear whether these tumors represent a homogeneous group with uniformly poor survival. This study identifies certain histologic subtypes of large cell and small cell undifferentiated carcinoma which have a better prognosis after curative resection than other similarly treated undifferentiated carcinomas. From 1947 through 1975, a total of 2,352 patients with lung cancer were admitted to one hospital. Follow-up to death was available in 98%. Pathological material was reviewed from 1,979 cases by a team of three pathologists during a single 6 month period without knowledge of clinical outcome. Curative resection was carried out in 632, with 170 (27%) 5 year survivors. Small cell cancer occurred in 481 patients and nine (1.6%) survived 5 years. Curative resections were performed in 34 with polygonal small cell carcinoma, 20 with normal lymph nodes and 14 with diseased lymph nodes. Seven survived 5 years (21%), six of 20 with normal and one of 14 with diseases lymph nodes. Eleven with nonpolygonal small cell carcinoma (eight oat cell, three fusiform) (five normal, six diseases nodes) underwent curative resection, with no survivors. Large cell carcinoma occurred in 151 and 19 survived 5 years. Curative resection was performed in 24 having large cell carcinoma with stratification (16 normal, eight diseased nodes), and 12 (50%) survived 5 years. In 26 with nonstratified large cell carcinoma undergoing curative resection (18 normal, eight diseases nodes); six (23%) survived 5 years (chi 2 = 4.06 p less than 0.05). Thus patients with resectable polygonal small cell carcinoma appear to have a better prognosis than those with nonpolygonal small cell carcinoma, and their prognosis approaches that of all patients with resectable lung cancer. Patients having resectable large cell carcinoma with stratification have a significantly better prognosis than those with nonstratified large cell carcinoma. Patients with these subtypes should therefore not be denied an attempt at curative resection because of the diagnosis of undifferentiated lung cancer. PMID- 6290803 TI - A reappraisal of limited-stage undifferentiated carcinoma of the lung. Does stage I small cell undifferentiated carcinoma exist? AB - Patients with small cell undifferentiated carcinoma of the lung (SCUC) have a poor prognosis. Surgical excision is avoided if the diagnosis can be made with small biopsy specimens or cytologic preparations. We reviewed 323 consecutive patients with pulmonary neoplasms diagnosed as SCUC, oat cel carcinoma, and undifferentiated or poorly differentiated carcinoma. At the time of diagnosis, only 18 patients had neoplasms classified as clinical Stage I, and only one of these had SCUC after histologic review. Fifteen patients had atypical carcinoid, a tumor with features intermediate between ordinary bronchial carcinoid and SCUC. In two instances, there was insufficient tissue for definitive diagnosis. Cumulative survival of the 15 patients with Stage I atypical carcinoid tumor was 80% at 1 year and 60% at most recent follow-up (mean follow-up 20 months). Mean survival for the 305 remaining patients was 7.9 months. Atypical carcinoid may be misdiagnosed as SCUC or poorly differentiated carcinoma, particularly with limited tissue samples or cytologic preparations. Stage I SCUC exists but is exceedingly rare. Many examples of purported Stage I SCUC probably represent atypical carcinoid. Because atypical carcinoid has a far better prognosis than SCUC, precise diagnosis is important and surgical resection should be considered. PMID- 6290804 TI - Melanotic neuroectodermal tumor of infancy: a malignant tumor of the femur. AB - An 18-month-old white girl had a malignant melanotic neuroectodermal tumor of infancy of the left femur. Light microscopic findings of a biopsy specimen of the femoral lesion demonstrated the characteristic histologic features, that is, nests of small cells with hyperchromatic nuclei and scant cytoplasm lying within cleftlike spaces lined by cuboidal, melanin-producing cells. Electron microscopy confirmed both neuroblastic and melanocytic differentiation. Despite radiation and chemotherapy, an extensive pelvic tumor developed, which prompted laparotomy 20 months after the femoral biopsy, The apparently metastatic tumor was largely necrotic, but viable nests of undifferentiated small cells and rare individual melanin-containing cells were identified. The patient died 4 months later, at 3 years of age. This case is of particular importance because it is the first example of this tumor found in a long bone. No similar lesions were seen in 17,000 primary tumors of bone treated and seen in consultation at this institution. Its aggressively malignant clinical course is also unusual; only 6 of the 159 cases of melanotic neuroectodermal tumor of infancy reported to date have expressed malignant behavior. PMID- 6290806 TI - Constipation and fecal impaction. PMID- 6290805 TI - Age-dependent alterations in lipids and function of rat heart sarcolemma. AB - The objective of the present study was to examine the effect of age on heart sarcolemma structure and function. Sarcolemmal fractions were prepared from hearts of young (1-1.5 months) and adult (10-12 months) rats and assayed for marker enzyme activities. The membrane fractions were found to be devoid of other cellular organelles upon examination by electron microscopy. They were enriched with 5'-nucleotidase and devoid of succinate dehydrogenase activity. The only age related lipid compositional changes noted in these membranes were changes in the fatty acid composition of membrane phospholipids with increasing age. Most changes were detected in phosphatidylcholine and phosphatidylethanolamine with very little alteration of sphingomyelin and phosphatidylserine plus phosphatidylinositol fatty acids. Polyunsaturated fatty acids, especially 18:2 and 20:4, were decreased with saturated fatty acids increased in membrane phosphatidylcholine and phosphatidylethanolamine fractions as the animal develops. There was a decrease in the specific activities of (Na+ + K+)-ATPase and 5'-nucleotidase of these membranes with age. On the other hand, membrane (K+) rho-nitrophenylphosphatase was not affected by age. PMID- 6290807 TI - Malignant hyperthermia. AB - The true incidence of malignant hyperthermia is unknown, but the frequency has been estimated as high as 1/14,000 anesthetic events. Review of the literature reports mortality rates up to 70%. Without prompt medical intervention, it is a uniformly fatal disease. Thus, it behooves the physician to have an awareness of the syndrome and its features, so that early recognition and adequate treatment take place. This paper presents a review of the literature on the occurrence, pathology, symptoms and treatment of malignant hyperthermia. PMID- 6290809 TI - [Receptors: from a pharmacological theory to the basis for medical thinking. 1. The biochemical reality of receptors]. PMID- 6290808 TI - [Double immunofluorescence detection of HBsAg and HBcAg in Councilman bodies of the liver]. AB - It could be shown by double immune fluorescence methods that HBsAg and/or HBcAg were present in Councilman bodies of the liver as well as in normal hepatocytes in biopsy specimens of a patient with HBsAg-seronegative chronic active hepatitis. These findings support earlier observations, that HBsAg is deposited in necrotic liver cells, and it supports the hypothesis, that Councilman bodies are formed under the influence of virus. PMID- 6290810 TI - Effects of phytohaemagglutinin, wheat-germ agglutinin, and concanavalin-A on the physical state of sialic acid and membrane proteins in human erythrocyte ghosts: a spin label study. PMID- 6290811 TI - Caffeine stimulates beta-endorphin release in blood but not in cerebrospinal fluid. AB - Plasma beta-endorphin and prolactin profiles were obtained from groups of unstressed, adult male rats. The infusion of caffeine (20 mg/kg) via a chronic, indwelling intra-atrial cannula results in a prompt and sustained (2-2.5 h) rise In plasma beta-endorphin levels. The infusion of the opiate antagonist naloxone causes a modest (40%) decrease in plasma beta-endorphin and blunts the elevation in plasma beta-endorphin following caffeine administration. In contrast, plasma prolactin levels were unchanged following caffeine administration and were decreased by treatment with naloxone. Caffeine treatment did not effect CSF beta endorphin levels or the release of beta-endorphin from hemipituitaries incubated in vitro. PMID- 6290812 TI - Domperidone elevates rat plasma beta-endorphin-immunoreactivity when administered peripherally but not intracerebroventricularly. AB - Domperidone, a dopamine receptor antagonist which apparently does not penetrate the blood-brain barrier in rats was administered to adult males. Domperidone 500 micrograms and 100 micrograms, given through intracarotid cannula, significantly elevated plasma beta-endorphin-immunoreactivity (beta-EP-I) at +15 min. To show that only a peripheral site(s) of action is implicated, domperidone was given to rats by cannulae implanted into both lateral ventricles. Plasma beta-EP-I was unaffected by this route of administration. These results suggest that plasma beta-EP-I is tonically inhibited by dopamine acting at site(s) outside of the blood-brain barrier. PMID- 6290813 TI - A characterization of beta-adrenergic receptors on cellular and perigranular membranes of rat peritoneal mast cells. AB - Beta-adrenergic receptors were characterized by measuring the specific binding of 3H-dihydroalprenolol (DHA) on intact isolated rat peritoneal mast cells (RPMC) and on perigranular membranes derived from purified RPMC granules. The specific binding of 3H-DHA reaches an equilibrium within 30 min at 5 degrees C and is linear with cell number. Scatchard analysis reveals two populations of binding sites on intact cells: with KD = 10.6 +/- 2.6 and 129 +/- 4.7 nM and Bmax of 186 +/- 38 and 1200 +/- 415 fmol/10(6) cells, respectively. Each cell contains 120 X 10(3) high-affinity binding sites and 720 X 10(3) low-affinity binding sites. There appears to be neither alpha-adrenergic nor muscarinic cholinergic receptors on the RPMC. Specific binding of 3H-DHA also occurred to isolated granules with perigranular membranes. The binding was saturable with a single population of binding sites with an affinity (KD) of 7.0 +/- 0.45 nM. Maximum binding (Bmax) was calculated at 56.6 +/- 1.9 fmol/10(9) granules. Subfractionation of granule components demonstrated that the specific binding sites appear to be localized exclusively on the perigranular membrane. PMID- 6290814 TI - Synthesis and biological properties of ovine corticotropin-releasing factor (CRF). AB - The 41-residue sequence of recently identified ovine corticotropin-releasing factor (CRF) was assembled on a benzhydrylamine resin support. Deprotection and cleavage from the resin were accomplished by HF treatment. The crude peptide was purified by gel filtration and reverse-phase, medium pressure, followed by high performance liquid chromatography (HPLC). In addition to the usual criteria, the homogeneity of the final material, obtained in 7% yield, was assessed by the isolation and examination of cyanogen bromide cleavage and tryptic digestion fragments by HPLC and amino acid analysis. The synthetic 41 amino acid CRF stimulated the release of corticotropin (ACTH) in three in vitro systems: isolated rat pituitary quarters, monolayer cultures of dispersed pituitary cells, and superfused pituitary cells on a column, the responses being related to the log-dose of CRF in the range of 0.05-125 ng/ml. The synthetic peptide also augmented in vivo release of ACTH in rats pretreated with chlorpromazine, morphine, and Nembutal, as assessed by the measurement of serum corticosterone. The data indicates chemical purity and high biological activity of synthetic material. PMID- 6290815 TI - Specific interaction among some enzymes and sodium dodecyl sulfate. AB - The effect of 1-butanesulfonic acid sodium salt and sodium dodecyl sulfate on the activity of highly purified and crystalline enzymes with marked differences in structure and function has been studied. The enzymes were: alcohol dehydrogenase; lactate dehydrogenase; malate dehydrogenase; isocitrate dehydrogenase; glucose-6 phosphate dehydrogenase; lipase; alkaline phosphatase. While 1-butanesulfonic acid sodium salt, at the studied concentrations, resulted generally inactive, sodium dedecyl sulfate showed a selective inhibitory effect, always under the critical micellar concentration. A kinetic analysis of the inhibitory action was also carried out. PMID- 6290817 TI - Cardiac beta-receptor variation in rat strains selectively bred for differences in susceptibility to stress. AB - The radioligand 3H-DHA was used to estimate the density and affinity of cardiac beta-receptors in rat strains selectively bred for differences in response to stress. Maudsley Reactive rats selected for heightened reactivity to stress had a greater density of beta-adrenergic binding in cardiac membranes than rats of two genetically distinct Maudsley Non-Reactive strains selected for decreased reactivity to stress, and compared with one of these Non-Reactive strains the MNR/Har, increased affinity for 3H-DHA. Together with previous findings the present results demonstrate a negative correlation between estimates of basal sympathetic activity on the on hand, and post-synaptic beta-receptors in heart on the other, that are consistent with the notion that these receptor alterations have occurred as a result of long-term differences in pre-synaptic release of transmitter. The Maudsley strains may, therefore, provide a useful model for the study of beta-adrenergic receptors as a physiological locus for regulation of end target responsiveness to sympathetic stimulation. PMID- 6290816 TI - Depolarizing agent-induced cyclic AMP accumulation in isolated rat spinal cord. AB - The stimulation of cyclic adenosine 3',5'-monophosphate (cyclic AMP) accumulation by the depolarizing agents K+, ouabain and veratridine, was studied in rat and guinea pig spinal cord tissue slices. Significantly increased accumulation of cyclic AMP was produced by each of the agents in a concentration-dependent manner. Veratridine and ouabain were equipotent (EC50 = 5 x 10(-5)M) and approximately 500 fold more potent than K+ (EC50 = 10(-2)M). Depolarizing agent induced cyclic AMP accumulation in slices from guinea pig spinal cord was approximately double the response in rat spinal cord. Maximum stimulation occurred within 2.5 min of incubation with these agents and lasted for at least 30 min. Regional studies demonstrated that the maximal accumulation of cyclic AMP occurred to a greater degree in tissue slices from the dorsal section of spinal cord from both rat and guinea pig. Whereas the ouabain and veratridine stimulatory responses are completely dependent on extracellular Ca++, the K+ response is only partially dependent. Stimulation due to ouabain and veratridine is dependent, and K+ is independent, of release of neurohumoral substances such as norepinephrine or adenosine from spinal neurons. These experiments indicate the possible modulatory role of depolarization-linked events in regulating the spinal cord cyclic AMP system. PMID- 6290818 TI - Discriminative stimulus properties of stereoisomers of cyclazocine in phencyclidine-trained squirrel monkeys. AB - Squirrel monkeys were trained to discriminate 0.16 mg/kg phencyclidine (PCP) from saline in a two-layer drug discrimination task on a fixed-ratio 32 schedule of food presentation. After reliable discriminative control of lever choice was established, dose-response determinations for generalization to the training dose of PCP were made with several doses of PCP, a racemic mixture of cyclazocine and the pure (+)- and (-)-isomers of cyclazocine. Only PCP and the (+)-isomer produced dose-dependent PCP-appropriate responding. Neither the racemic mixture nor (-)-cyclazocine produced over 25% PCP-appropriate responding at any of the doses tested. (+)-Cyclazocine was eight times less potent than PCP in producing drug-lever appropriate responding. (-)-Cyclazocine was about 30 times more potent than PCP and over 200 times more potent than (+)-cyclazocine in overall response rate suppression. The potency of the racemic mixture for response-rate suppression was consistent with an additive effect of the isomers. Neither the PCP-lever appropriate responding produced by (+)-cyclazocine nor the response rate suppression produced by (-)-cyclazocine were antagonized by naloxone. Thus, racemic cyclazocine is composed of two isomers with differing behavioral effects. The (-)-isomer is more potent, and the (+)-isomer has more specificity for PCP like effects. PMID- 6290820 TI - Current concepts: II. Hormonal regulation of molecular events during aging. AB - This review is an attempt to establish a role for both glucocorticoid and thyroid hormones in the aging process as factors which mediate the regulation of transcription and translation of specific genetic domains. Furthermore, through a review of the current knowledge of the effect of long-term hypophysectomy on physiological and biochemical functions which change with age in the rat an attempt has been made to establish a role for a pituitary factor in the regulation of the aging process through an affect on transcription and translation of hormonally induced genetic domains. Although we can propose a possible mechanism of action of this factor, the proposal is premature and requires extensive systematic testing. It will be interesting to carefully consider the possible relationship, if any, of this pituitary factor to the effects of dietary restriction on animal longevity. How this pituitary-aging interaction might be exploited is open to speculation but it is certainly worth further consideration based on evidence currently at hand. PMID- 6290819 TI - Dopamine accumulation after dopamine beta-hydroxylase inhibition in rat heart as an index of norepinephrine turnover. AB - Dopamine concentration in rat heart is normally very low, only a few percent of the concentration of norepinephrine. After treatment of rats with a dopamine beta hydroxylase inhibitor, 1-cyclohexyl-2-mercapto-imidazole (CHMI), there was a rapid increase in dopamine concentration even before norepinephrine concentration had decreased perceptibility. This accumulation of dopamine was readily measured by liquid chromatography with electrochemical detection. Since the percentage change in dopamine was much greater than the percentage change in norepinephrine, especially at early times, measurement of dopamine accumulation rather than norepinephrine decline was considered as a useful measure of norepinephrine turnover. Drugs that act on noradrenergic receptors and are known to alter norepinephrine turnover were found to alter the rate of dopamine accumulation. Clonidine and guanabenz decreased dopamine accumulation after CHMI, whereas piperoxan (but not prazosin) increased dopamine accumulation after CHMI. Pergolide, a dopamine agonist whose lowering of blood pressure and cardiac rate has been suggested to be due to suppression of neurogenic release or norepinephrine, also decreased dopamine accumulation after CHMI. The results suggest that measuring dopamine accumulation may have advantages over measuring norepinephrine disappearance after dopamine beta-hydroxylase inhibition as an indicator of norepinephrine turnover in heart. PMID- 6290821 TI - Litorin suppresses food intake in rats. AB - Litorin (LIT), a bombesin-like nonapeptide, decreased food intake in rats in a dose-related manner after parenteral injection. LIT decreased deprivation-induced water intake only at a dose much higher than required to suppress feeding. LIT administration did not significantly alter the frequency of observed feeding associated behaviors, nor did it result in subsequent aversion to an associated novel solution. Litorin shares with bombesin structural features and pharmacological actions that include the suppression of food intake in a manner that mimics natural satiation. PMID- 6290822 TI - Sleep: sequential reduction of paradoxical (REM) and elevation of slow-wave (NREM) sleep by a non-convulsive dose of insulin in rats. AB - Administration of a single non-convulsive dose of insulin (1.0, I.U./kg., I.P.) which produced no observable gross behavioral changes in rats, reduced rapid eye movement (REM) sleep time 100% in the first 3 hrs. and 82% by the 4th hr., reaching control subject levels (saline-treated) by the 6th hr. In contrast, slow wave sleep (NREM) time in insulin treated animals exceeded control subject levels by 49% by the end of the 2nd hr., returning to normal by the 5th hr. Although there was no difference between insulin and saline treated rats for the total 8 hr. post-injection recording period for total percentage of time awake, or slow wave sleep time, a 44% reduction in REM sleep time was observed in insulin treated animals compared to that of a saline treated control. The significance of these findings are discussed in terms of known neurochemical changes i.e., an increase of both brain tryptophan and serotonin in rats, induced by a subconvulsive dose of exogenous insulin. PMID- 6290823 TI - In vitro recording of raphe unit activity: evidence for endogenous rhythms in presumed serotonergic neurons. AB - The spontaneous activity of single neurons in the nucleus raphe dorsalis was recorded in vitro in mouse brain slices. The neurons displayed the slow and regular discharge pattern characteristic of raphe neurons recorded in vivo. When magnesium ion was added to increase the medium concentration to 20-30 mM for the purpose of inhibiting all synaptic transmission, raphe neurons continued to display the same discharge pattern and rate. The data suggest that the steady rhythmic firing of nucleus raphe dorsalis neurons is generated by an intracellular pacemaker mechanism. PMID- 6290824 TI - Hormonal regulation of beta-adrenergic receptors in fetal rabbit lung in organ culture. AB - Explants of fetal rabbit lung were established on the 25th day of gestation. These were maintained in serum-free medium for periods up to 10 days. During this time, the cultures exhibited morphological changes typical of terminal lung differentiation. Morphological evidence was also obtained for synthesis and secretion of pulmonary surfactant in these explants. beta-Adrenergic receptors were identified in these lung explants. Exposure of the explants to 10(-7)M dexamethasone on the third day of culture resulted in a significant increase in the number of beta-adrenergic receptors in the tissue without a change in receptor affinity. The effect of dexamethasone in organ culture was dose dependent, a maximum increase in receptor number being observed within 48 hours of incubation with a hormone concentration of 1 x 10(-7)M. Exposure of the explant tissue to 1 x 10(-7)M triiodothyronine resulted in no significant increase in the concentration of beta-adrenergic receptors and no change in receptor affinity. These results suggest that glucocorticoids may potentiate the effects of beta-adrenergic agents in the fetal lung by increasing the numbers of their receptors. The effects of triiodothyronine upon the fetal lung do not appear to be mediated by this mechanism. PMID- 6290825 TI - Diseases of the fetus and neonate due to human cytomegalovirus: a laboratory perspective. PMID- 6290826 TI - [Radiotherapy of chemodectomas of the neck]. PMID- 6290827 TI - [Computer processing of radionuclide scintigrams for the diagnosis of acute myocardial infarct]. PMID- 6290828 TI - [Hormones and ferritin in children with lymphogranulomatosis]. PMID- 6290829 TI - Apparent suicide by carbon monoxide poisoning in a case of insulinoma. PMID- 6290830 TI - [Multimorbidity of cancer patients. Study of 480 cancer patients]. PMID- 6290831 TI - [Experiences with the oral antimycotic ketoconazole in vaginal fungal involvement]. PMID- 6290832 TI - [Response of calomys musculinus to experimental infection with Junin virus]. PMID- 6290833 TI - Stimulation of cortical bone mineralization and remodeling by phosphate and 1,25 dihydroxyvitamin D in vitamin D-resistant rickets. AB - Besides rachitic and osteomalacic bone lesions specific disturbances of intracortical bone remodeling have been described in children with vitamin D resistant rickets (VDRR). The effects of phosphate and 1,25-dihydroxyvitamin D3 [Pi + 1,25(OH)2D] on the abnormal cortical bone remodeling were assessed by static and dynamic histomorphometric analysis of dual labeled undecalcified iliac crest bone biopsies obtained from 12 young VDRR children. Bone mineralization was markedly improved as shown by reduction of the osteoid thickness, shortening of the mineralization lag time and of the osteon calcification period. In conjunction with improved bone mineralization the extent of dual labeled bone surface was increased together with the osteoblast population, indicating that normal bone calcification requires the presence of osteoblasts. At the tissue level the birthrate of new Basic Multicellular remodeling Units (BMU) was clearly enhanced; while at the cellular level, the low calcification rate remained unchanged in most cases. The data show that treatment with Pi + 1,25(OH)2D stimulates the bone turnover in young patients with VDRR by inducing creation of new BMU after restoration of bone mineralization. Unlike the increased recruitment of new BMU caused by treatment, the persistence of a low calcification rate may reflect the existence of a primary osteoblast defect in some VDRR patients. PMID- 6290834 TI - A case of cataract formation during the lactating period associated with galactose-1-phosphate uridyl transferase deficiency. PMID- 6290835 TI - Modulation of insulin secretion by cyclic AMP and prostaglandin E: the effects of theophylline, sodium salicylate and tolbutamide. PMID- 6290836 TI - Evidence for independent actions of vasopressin on renal inner medullary cyclic AMP and prostaglandin E production: relationship of the prostaglandin E response to hormone pressor activity. AB - Arginine vasopressin (AVP) has been shown to stimulate prostaglandin (PG) production in renal medulla, while PGs have been implicated in the suppression of the antidiuretic activity of AVP. These findings have suggested a local negative feedback system involving PGs in the modulation of the antidiuretic activity of AVP. However, coupling of the antidiuretic activity of AVP to its action to increase medullary PG production is not established. In the present study of rat inner medullary slices, we concurrently examined in the same incubate the relationship between the actions of AVP to increase media immunoreactive PGE (iPGE) and tissue cAMP, the presumed first biochemical step in expression of the antidiuretic activity of the hormone. The synthetic AVP analogue [1, d(CH2)5Tyr(Me)AVP], which selectively blocks the pressor but not the antidiuretic activity of AVP, abolished the action of AVP to increase media iPGE in inner medullary incubates but did not alter AVP induced increases in tissue cAMP in slices from the same incubates. By contrast, the analogue [d(CH2)5Tyr(Et)VAVP], which blocks both the pressor and antidiuretic activity of AVP, inhibited both the cAMP and iPGE responses to AVP. The analogue 1-deamino-8-D-AVP (dD'AVP), which has potent antidiuretic activity but little if any pressor activity, markedly stimulated inner medullary cAMP accumulation without altering media iPGE. These results indicate that the acute actions of AVP to increase inner medullary cAMP and iPGE are separable and independent. The latter effect of AVP appears to be linked to the pressor rather than the antidiuretic activity of the hormone.U PMID- 6290837 TI - Phenformin has opposite effects on insulin and growth hormone binding to IM-9 lymphocytes. AB - We studied simultaneously the effect of various concentrations of phenformin on insulin and growth hormone binding to IM-9 lymphocytes, a cell type known to have receptors for both these hormones. After 24 hr preincubation with phenformin at 2 x 10(-5) M, insulin binding to IM-9 cells was increased by 80.4 +/- 10.5% over control (mean +/- SE of 10 experiments). In parallel experiments HGH binding was decreased by 43.1 +/- 2.2% (mean +/- SE). This effect of phenformin was dose dependent for both HGH and insulin binding over the concentration range 1.5 x 10( 6) M to 5 x 10(-5) M, and was already detectable 3 hr after phenformin addition. These data indicate that phenformin has an opposite effect on insulin and growth hormone binding to IM-9 cells. Several possible mechanisms might be suggested for the decrease of HGH binding sites induced by phenformin: the simultaneous opposite effect on HGH and insulin receptors raises the possibility that some metabolic event triggered by the drug is able to induce opposite changes in the binding of these two hormones with different biological activities. PMID- 6290838 TI - Obesity and the regulation of phosphofructokinase in heart: an apparent insensitivity to adrenergic activation in mature-age genetically obese rats. AB - The activity ratio of phosphofructokinase in perfused rat heart and its activation by epinephrine was examined in non-obese, fat-fed obese, and genetically obese rats. For non-obese colony rats there was an age-dependent increase in the activity ratio of phosphofructokinase from 0.2 at 40 days to 0.4 at mature age (greater than 200 days). Epinephrine (10 microM) treatment of the heart for 5 min increased the ratio at all ages but the proportional increase diminished with age. For mature-age lean Zucker rats carrying the genetic determinant for obesity the results were similar to those obtained for comparable non-obese colony rats. For fat-fed obese rats the activity ratio of phosphofructokinase at 200 days of age was 0.2 and was increased to 0.6 by epinephrine treatment. For mature-age obese Zucker rats the activity ratio was 0.2 and no significant response to epinephrine occurred. The activity ratio of glycogen phosphorylase and its response to epinephrine (beta-adrenergic receptor mediated) in heart was unaffected by age, diet or the gene for obesity. The present findings indicate a specific defect in the adrenergic regulatory mechanism for phosphofructokinase in genetically obese rats. PMID- 6290839 TI - Purification and assay of 15-ketoprostaglandin delta 13-reductase from bovine lung. PMID- 6290840 TI - Receptors for PGI2 and PGD2 on human platelets. PMID- 6290841 TI - Distribution of PGE and PGF2 alpha receptor proteins in the intracellular organelles of bovine corpora lutea. PMID- 6290842 TI - A receptor for prostaglandin F2 alpha from corpora lutea. PMID- 6290843 TI - Purification and characterization of leukotrienes from mastocytoma cells. PMID- 6290844 TI - Rapid extraction of arachidonic acid metabolites from biological samples using octadecylsilyl silica. PMID- 6290845 TI - Prenatal analysis of human DNA-sequence variation. PMID- 6290848 TI - Changed properties of the A subunit in DNA gyrase with a B subunit mutation. AB - To investigate the interaction of subunits A and B of DNA gyrase during DNA supercoiling, a Cour mutant of Escherichia coli was obtained and the effect of nalidixic acid on the supercoiling of DNA by wild-type and mutant enzymes was assayed. The enzyme of the Cour strain proved to be more sensitive to nalidixic acid than the wild-type DNA gyrase. Hence the mutation affecting the B subunit can also change the properties of the A subunit, which fact suggests that the two subunits of DNA gyrase are in contact during DNA supercoiling. PMID- 6290846 TI - Negative control of ornithine decarboxylase and arginine decarboxylase by adenosine-3':5'-cyclic monophosphate in Escherichia coli. AB - The polyamine biosynthetic enzymes, ornithine decarboxylase (EC 4.1.1.17) (ODC) and arginine decarboxylase (EC 4.1.1.19) (ADC), are negatively controlled by cAMP in Escherichia coli. The specific activities of ODC and ADC were determined in crude extracts prepared from E. coli strains carrying a mutation in the adenylate cyclase (EC 4.6.1.1) structural gene (cya) and wildtype strains. These strains were cultured on various carbon sources in the presence and absence of cAMP. In wild-type strains, ODC and ADC activities were diminished in cells grown on glycerol compared to these strains cultured on glucose. When cya strains were grown on glucose or glycerol, ODC and ADC activities were the same. Addition of 1 mM cAMP to glucose-based medium repressed ODC and ADC activities in both the wild type and cya strains. Furthermore, cAMP exerts its negative control through the cAMP receptor protein, since strains carrying a mutation in the crp structural gene fail to repress ODC and ADC activities in response to increased cAMP obtained by carbon source manipulation or cAMP supplementation of the growth medium. This evidence suggests that negative control of ODC and ADC by cAMP occurs at the level of transcription. PMID- 6290847 TI - The Escherichia coli dnaW mutation is an allele of the adk gene. AB - A dnaW mutant, isolated on the basis of inability to effect conjugal DNA transfer at high temperature, has been shown by complementation and enzyme assay to be defective in the adk (adenylate kinase; EC 2.7.4.3) locus. The adk mutant, known to have reduced ATP concentration at the nonpermissive temperature (Cousin and Belaich 1966), was used to demonstrate a donor energy requirement for stable aggregate formation and for chromosome transfer in conjugation. PMID- 6290849 TI - Binding of HBs antigen to HeLa cells by use of reconstitution into liposomes and fusion by Sendai virus. AB - Hepatitis B surface antigen (HBsAg) was reconstituted into liposomes composed of phosphatidylserin. An isolated membrane fraction of HeLa cells was mixed with the liposomes, then the liposomes were incubated with HeLa cells in the presence of Sendai virus. In this system the attachment of HBsAg to the cell surface was enhanced and became resistant to treatment with trypsin-EDTA. The presence of the HBsAg on the cell surface was revealed by immunoelectronmicroscopy. This technique may provide a model system for studying immunological reactions to HBsAg-bearing target cells in vitro. PMID- 6290850 TI - Mechanism of antibody-mediated protection against herpes simplex virus infection in athymic nude mice: requirement of Fc portion of antibody. AB - Experiments were performed to investigate the resistance of the host due to antibody-mediated mechanisms to herpes simplex virus (HSV) infection. Transfer of hyperimmune anti-HSV mouse serum inhibited the development of skin lesions and prolonged the survival of lethally HSV-infected nude mice. Relatively high concentrations of antibody were required to achieve this protection. Antisera prepared in heterologous animals were also effective, while administration of anti-cowpox virus serum or interferon provided no protection. This type of protection is therefore due to specific antibody and cannot be attributed to interferon. In order to delineate the requirement for antibody in antibody mediated protection, human gamma globulin preparations were transferred to lethally HSV-infected nude mice. Transfer of intact human gamma globulin (GG) was effective in controlling infection. S-sulfonation of GG did not diminish the protective ability. However, purified F(ab')2 did not have any protective action even when it was administered frequently to maintain serum neutralizing antibody titer. GG was effective in C5-deficient mice lethally infected with HSV. These results indicate that in vivo antibody-mediated protection to HSV infection requires the Fc region of the intact IgG molecule and suggest that antibody dependent cell-mediated cytotoxicity may be operative in vivo. PMID- 6290851 TI - Interferon induction by glycyrrhizin and glycyrrhetinic acid in mice. PMID- 6290852 TI - Rapid identification and typing of herpes simplex virus in clinical specimens by a direct microneutralization test. PMID- 6290853 TI - Expression of target antigen for Epstein-Barr virus-specific cytotoxic T cells on BJAB cells freshly infected with EBV. AB - The target antigen for Epstein-Barr virus (EBV)-specific cytotoxic T cells (Tc) was expressed on BJAB cells exposed to the B95-8 strain of EBV for at least one hour. Ultraviolet-light (UV)-irradiated noninfectious B95-8 virus also induced the target antigen on BJAB cells. Cold target competition tests suggested that the target antigen expressed on EBV-infected BJAB cells was distinct from the lymphocyte-detected membrane antigen (LYDMA) which was also recognized by the EBV specific Tc and expressed on autologous EBV-Transformed lymphoblastoid cell line (LCL) cells. Neither of these target antigens for EBV- specific Tc was detected on the surface of EBV-genome positive BJAB cells which had been kept in a long term culture after EBV-infection. Thus, the virion antigen, especially the EBV membrane antigen (MA), is a possible candidate for the target antigen expressed on EBV-infected BJAB cells. Lysis of EBV-infected BJAB cells was inhibited by target cell treatment with anti-beta 2 microglobulin (anti-beta 2M) antibody and induction of the effector Tc was dependent on the donor individual. These results suggested the possibility that the Tc recognizing EBV-infected BJAB cells are restricted by the major histocompatibility complex (MHC). PMID- 6290854 TI - In vitro activity of cefotaxime, ceftizoxime, cefmenoxime, and latamoxef in comparison with other beta-lactam antibiotics against recent clinical isolates of Haemophilus influenzae and Haemophilus parainfluenzae. PMID- 6290855 TI - [Products of the enzymatic hydrolysis of collagen by Bacillus subtilis proteases]. PMID- 6290856 TI - [Survival of melanin-containing fungi under superhigh vacuum]. PMID- 6290857 TI - [Sensitivity of bentonite-immobilized enteroviruses to organic solvents]. PMID- 6290858 TI - Cytochrome c reduction by triiodothyronine (T3). AB - Triiodothyronine (T3), the active thyroid hormone, reduced cytochrome c non enzymatically. This reduction was partially inhibited by superoxide dismutase (SOD). Thyroxine (T4) was only minimally effective as a reductant in this system. The effect of T3 on mitochondrial oxidative phosphorylation may involve a direct redox mechanism. PMID- 6290859 TI - Viral hepatitis: a four-year hospital and general-practice study in Sydney 1. Epidemiological features, natural history, and laboratory findings. AB - We studied 761 patients admitted to hospital with viral hepatitis between 1971 and 1974, and 53 patients with viral hepatitis seen in general practice in Sydney, following up some of them for one to two years. We evaluated factors contributing to each type of hepatitis. We noted differences in the patterns in hepatitis A, B and non-A non-B between Anglo-Saxon and non-Anglo-Saxon sectors of the community. All patients with hepatitis A regained normal liver function within 20 months of the acute illness. Of 115 hepatitis B patients seen at 12 months, 6% had chronic hepatitis Bs antigenaemia, 60% had developed anti-HBs antibodies, and 7.3% still had abnormal liver function. Of 20 non-A non-B patients followed for 12 months, liver function was still abnormal in three, but one of these had developed hepatitis B. The case fatality rate for the whole series was 0.66%. PMID- 6290860 TI - Skeletal muscle biopsy: indications and results in 200 patients. PMID- 6290861 TI - [Effect of a fiber-rich diet on digoxin resorption]. AB - In five female healthy volunteers the influence of dietary fiber (wheat bran or carob seed flour) on absorption of digoxin was investigated. Five minutes after ingestion of a formula diet alone or in combination with wheat bran or carob seed flour 0,8 mg beta-acetyldigoxin was given per os. The plasmaconcentration-time curve over eight hours, the area under curve and the cumulative urinary excretion were not changed significantly. It was concluded that there is no influence of dietary fiber on rate or degree of digoxin-absorption. PMID- 6290862 TI - Streptozocin (Zanosar). PMID- 6290863 TI - Triple-barrelled ion-sensitive microelectrode for simultaneous measurements of two extracellular ion activities. AB - A method has been described by which triple-barrelled ion-sensitive microelectrodes for simultaneous measurements of two extracellular ion activities could be produced with a tip diameter of 1-2 microns. The following combinations with two ion exchangers could be achieved: Ca++/K+, Ca++/Cl-, K+/Na+. The mean steepness for the potassium electrode was 55.8 mV, for the calcium electrode 26 mV, for the chloride electrode 48.8 mV, and for the sodium electrode 55.2 mV. The change of steepness varied 4%-5%/day and some of the electrodes could be used up to 5 days. The mean response time for all electrodes did not amount to more than 200 ms. An example is given for simultaneous in vivo measurement of both extracellular potassium and calcium activity. PMID- 6290865 TI - Inhibition of prostaglandin synthesis antagonizes the colchicine-induced reduction of vasopressin-stimulated water flow in the toad urinary bladder. PMID- 6290866 TI - Muscarinic cholinergic receptors on cultured thyroid cells. I. Biological effect of carbachol and characterization of the receptors. AB - A muscarinic cholinergic effect on thyrotropin-stimulated cyclic AMP accumulation in cultured porcine thyroid cells is characterized. The muscarinic agonists carbachol, acetylcholine, oxotremorine, and pilocarpine decreased the acute cyclic AMP response to stimulation with thyrotropin (20 mU/ml). A 50% decrease was obtained in the presence of 0.1 mM carbachol. Maximal effects were observed when cells were cultured in the presence of thyrotropin (100 microU/ml) or prostaglandin E2 (1 microM), whereas cells cultured in basal medium or in the presence of dibutyryl cyclic AMP showed a low response to carbachol. Evidence is reported suggesting that carbachol acutely decreases cyclic AMP synthesis. The properties of the muscarinic receptors present in thyroid cells were defined by using the binding of the muscarinic antagonist [3H]quinuclidinyl benzilate to cell homogenates. Scatchard plots revealed a single population of binding sites with a KD of 0.3 nM. The numbers of binding sites per cell after 4 days in culture were 880 in control cells, 4335 in thyrotropin (100 microU/ml)-treated cells, 5600 in prostaglandin (1 microM)-treated cells, and 1420 in dibutyryl cyclic AMP-treated cells. Therefore the sensitivity to carbachol appeared to be related to the number of antagonist binding sites and to be independent of the sensitivity of the cells to acute thyrotropin stimulation. PMID- 6290864 TI - Similarities of lipid metabolism in mammalian and protozoan cells: an evolutionary hypothesis for the prevalence of atheroma. PMID- 6290867 TI - Muscarinic cholinergic receptors on cultured thyroid cells. II. Carbachol-induced desensitization. AB - Muscarinic cholinergic receptors were previously characterized on cultured porcine thyroid cells. The receptor number was increased by chronic thyrotropin or prostaglandin E2 treatments [Champion, S., and J. Mauchamp. Mol. Pharmacol. 21:66-72 (1982)]. The long-term effect of carbachol was studied. After chronic treatment with carbachol, cells were completely desensitized to acute carbachol stimulation. This process was blocked by muscarinic antagonists. A complete desensitization was obtained after 6 hr of treatment with 100 microM agonist. Under these conditions the quinuclidinyl benzilate binding capacity of cell homogenates was reduced by 50%. Withdrawal of carbachol allowed the complete restoration of the sensitivity of cells within 6 hr with only a partial recovery of the binding capacity (25%). The complete complement of receptors was obtained after 24 hr of recovery. Desensitization and agonist-induced decrease in receptor number were not affected by cycloheximide, whereas the recovery of both effect and binding sites required active protein synthesis. PMID- 6290868 TI - Interaction of beta-endorphin and other opioid peptides with calmodulin. AB - A highly purified preparation of calmodulin activated a calmodulin-deficient phosphodiesterase by more than 10-fold. This activation of phosphodiesterase by calmodulin was completely inhibited by two opioid peptides, beta-endorphin and dynorphin, at concentrations that had no appreciable effect on the basal phosphodiesterase activity. By contrast, similar concentrations of other structurally related peptides, including alpha-endorphin, (des-Tyr1)-gamma endorphin, Leu-enkephalin, and Met-enkephalin, failed to block calmodulin's activation of phosphodiesterase. The inhibition by beta-endorphin of calmodulin's action was not reversed by calcium or by the opiate antagonist naloxone but was overcome by increasing the concentration of calmodulin. Equilibrium dialysis studies showed that 125I-labeled beta-endorphin bound directly to calmodulin in a saturable, calcium-dependent manner with a dissociation constant of approximately 4.6 microM. There was substantially less binding of beta-endorphin to troponin-C and little or no calcium-dependent binding of beta-endorphin to bovine serum albumin, lactalbumin, or histone. This interaction of beta-endorphin with calmodulin was similar in several respects to the interaction of certain antipsychotic drugs to calmodulin and may explain certain of the peptide's biochemical effects. PMID- 6290869 TI - Rearrangements and deletions of immunoglobulin heavy chain genes in the double producing B cell lymphoma I.29. AB - The B cell lymphoma I.29 consists of a mixture of cells expressing membrane-bound immunoglobulin M (IgM) (lambda) and IgA (lambda) of identical idiotypes. Whereas most of the cells express either IgM or IgA alone, 1 to 5% of the cells in this tumor express IgM and IgA simultaneously within the cytoplasm and on the cell membrane (R. Sitia et al., J. Immunol. 127:1388-1394, 1981; R. Sitia, unpublished data). When IgM+ cells are purified from the lymphoma and passaged in mice or cultured, a portion of the cells convert to IgA+. These properties suggest that some cells of the I.29 lymphoma may undergo immunoglobulin heavy chain switching, although it is also possible that the mixed population was derived by a prior switching event in a clone of cells. We performed Southern blotting experiments on genomic DNAs isolated from populations of I.29 cells containing variable proportions of IgM+ and IgA+ cells and on a number of cell lines derived from the lymphoma. The results were consistent with the deletion model for heavy chain switching, as the IgM+ cells contained rearranged mu genes and alpha genes in the germ line configuration on both the expressed and nonexpressed heavy chain chromosomes, whereas the IgA+ cells had deleted both mu genes and contained one rearranged and one germ line alpha gene. In addition, segments of DNA located within the intervening sequence 5' to the mu gene, near the site of switch recombination, were deleted from both the expressed and the nonexpressed chromosomes. Although mu genes were deleted from both chromosomes in the IgA+ cells, the sites of DNA recombination differed on the two chromosomes. On the expressed chromosome, Smu sequences were recombined with S alpha sequences, whereas on the nonexpressed chromosome, Smu sequences were recombined with S gamma 3 sequences. PMID- 6290870 TI - Human gene (c-fes) related to the onc sequences of Snyder-Theilen feline sarcoma virus. AB - The onc gene (v-fes) of the acutely transforming feline sarcoma virus (Snyder Theilen strain) has homologous cellular sequences (c-fes) in all vertebrate species, including humans. We isolated from a human DNA library recombinant phages containing overlapping c-fes sequences. The human c-fes locus spans a region of 3.4 kilobases and contains 1.4 kilobases of DNA homologous to the viral onc sequence interspersed with three intervening sequences. PMID- 6290872 TI - Saccharomyces carlsbergensis fdp mutant and futile cycling of fructose 6 phosphate. AB - In Saccharomyces, the addition of glucose to cells grown in media lacking sugars causes irreversible inactivation of fructose bisphosphatase. One function of this process might be to prevent a futile cycle of formation and hydrolysis of fructose 1,6-bisphosphate. We tested such cycling by assessing the labeling of the 1-position of glucose in polysaccharides from [6-14C]glucose (J.P. Chambost and D. G. Fraenkel, J. Biol. Chem. 225:2867-2869, 1980) by using mutants impaired in glucose growth and known not to inactivate the phosphatase normally (i.e., the fdp mutant of Saccharomyces carlsbergensis [van de Poll et al., J. Bacteriol. 117:965-970, 1974] and the similar cif mutant of Saccharomyces cerevisiae [Navon et al., Biochemistry 18:4487-4499, 1979] ), as well as in the wild-type strain tested in the 1-h period before inactivation is complete. There was marginal, if any, cycling in any situation, and we conclude that the phosphatase activity is controlled by means other than inactivation or that the extent of cycling is too low to be significant, or both. For the fdp mutant data are also presented on growth, rate of glucose metabolism, metabolite accumulations, enzyme levels, and glucose transport, but the primary lesion is unknown. PMID- 6290873 TI - Mutant strains of Tetrahymena thermophila defective in thymidine kinase activity: biochemical and genetic characterization. AB - Three mutant strains, one conditional, of Tetrahymena thermophila were defective in thymidine phosphorylating activity in vivo and in thymidine kinase activity in vitro. Nucleoside phosphotransferase activity in mutant cell extracts approached wild-type levels, suggesting that thymidine kinase is responsible for most, if not all, thymidine phosphorylation in vivo. Thymidine kinase activity in extracts of the conditional mutant strain was deficient when the cells were grown or assayed or both at the permissive temperature, implying a structural enzyme defect. Analysis of the reaction products from in vitro assays with partially purified enzymes showed that phosphorylation by thymidine kinase and nucleoside phosphotransferase occurred at the 5' position. Genetic analyses showed that the mutant phenotype was recessive and that mutations in each of the three mutant strains did not complement, suggesting allelism. PMID- 6290871 TI - Two structurally and functionally different forms of the transforming protein of PRC II avian sarcoma virus. AB - The primary translation product of the PRC II avian sarcoma virus genome is a protein of 105,000 daltons (P105), and we have previously shown that approximately 50% of the P105 molecules are converted to molecules of 110,000 daltons (P110) by posttranslational modification. Fractionation of PRC II infected cells showed that P105 was contained primarily in a nonionic detergent soluble compartment, whereas P110 partitioned almost exclusively with a nonionic detergent-insoluble or crude cytoskeletal fraction. The tyrosine-specific protein kinase activity previously observed in immunoprecipitates which presumably contained both P110 and P105 was found predominantly in the P110-containing immunoprecipitates made from the cytoskeletal fraction and was essentially absent from the P105-containing immunoprecipitates prepared from the soluble fraction. Individual analysis of 32P-labeled P110 and P105 prepared by this fractionation technique revealed that P110 contained more phosphotyrosine per mole of protein than did P105. Examination of the tryptic peptide maps of 32P-labeled P110 and P105 suggested that the additional phosphotyrosine in P110 resulted from phosphorylation at discrete sites within the protein. From these experiments, we conclude that PRC II-infected cells contain two discrete forms, P105 and P110, of the transforming protein and that each of these proteins exhibits distinct structural and functional characteristics. PMID- 6290875 TI - Introduction and recovery of a selectable bacterial gene from the genome of mammalian cells. AB - The simian virus 40 (SV40)-pBR322 recombinant, pSV2, carrying the origin of SV40 replication and the gpt gene of Escherichia coli, has been stably introduced into Chinese hamster ovary hprt- cells. All gpt-transformed cell lines were found to contain one or more insertions of pSV2 sequences exclusively associated with high molecular-weight DNA. Additional analyses showed that at least one integrated copy in each cell line retained an intact gpt gene and flanking SV40 sequences required for expression of xanthine-guanine phosphoribosyltransferase. Most cell lines contained pSV2 sequences which had integrated with partial sequence duplication. Upon fusion with COS-1 cells, a simian cell line permissive for autonomous pSV2 replication, most gpt-transformed cell lines produced low molecular-weight DNA molecules related to pSV2. The majority of these replicating DNAs were indistinguishable from the original transfecting plasmid in both size and restriction enzyme cleavage pattern. In addition, the recovered DNA molecules were able to confer ampicillin resistance to E. coli and to transform mouse L cells and Gpt- E. coli to a Gpt+ phenotype. These studies indicate that all of the genetic information carried by this SV40-plasmid recombinant can be introduced into and retrieved from the genome of mammalian cells. PMID- 6290874 TI - Isolation and characterization of human DNA fragments with nucleotide sequence homologies with the simian virus 40 regulatory region. AB - A recombinant library of human DNA sequences was screened with a segment of simian virus 40 (SV40) DNA that spans the viral origin of replication. One hundred and fifty phage were isolated that hybridized to this probe. Restriction enzyme and hybridization analyses indicated that these sequences were partially homologous to one another. Direct DNA sequencing of two such SV40-hybridizing segments indicated that this was not a highly conserved family of sequences, but rather a set of DNA fragments that contained repetitive regions of high guanine plus cytosine content. These sequences were not members of the previously described Alu family of repeats and hybridized to SV40 DNA more strongly than do Alu family members. Computer analyses showed that the human DNA segments contained multiple homologies with sequences throughout the SV40 origin region, although sequences on the late side of the viral origin contained the strongest cross-hybridizing sequences. Because of the number and complexity of the matches detected, we could not determine unambiguously which of the many possible heteroduplexes between these DNAs was thermodynamically most favored. No hybridization of these human DNA sequences to any other segment of the SV40 genome was detected. In contrast, the human DNA segments isolated cross hybridized with many sequences within the human genome. We tested for the presence of several functional domains on two of these human DNA fragments. One SV40-hybridizing fragment, SVCR29, contained a sequence which enhanced the efficiency of thymidine kinase transformation in human cells by approximately 20 fold. This effect was seen in an orientation-independent manner when the sequence was present at the 3' end of the chicken thymidine kinase gene. We propose that this segment of DNA contains a sequence analogous to the 72-base-pair repeats of SV40. The existence of such an "activator" element in cellular DNA raises the possibility that families of these sequences may exist in the mammalian genome. PMID- 6290876 TI - Cloning and transcriptional control of a eucaryotic permease gene. AB - The uracil permease gene of the yeast Saccharomyces cerevisiae was cloned on a hybrid plasmid which replicates autonomously in both yeast and Escherichia coli. Cloning was carried out by complementation in yeast. The smallest DNA fragment found to complement the uracil permease deficiency in recipient yeast cells measured approximately 2.3 kilobases. In strains transformed by the plasmid with the uracil permease gene inserted, initial rates of uracil uptake increased up to 25 times more than the rates found in the wild type. Using DNA probes carrying several regions of the cloned gene, I showed that a strain carrying the dhul-I mutation, which is not linked to the permease structural gene and is responsible for enhanced uptake velocity of uracil, had enhanced transcription of the permease gene. By using DNA probes recloned in phage M13 mp7, the direction of transcription of the permease gene relative to the restriction map was deduced. A half-life of 2 min was found for the permease mRNA in labeling kinetics experiments. PMID- 6290877 TI - Covalent guanylyl intermediate formed by HeLa cell mRNA capping enzyme. AB - Guanylyltransferase, an enzyme that catalyzes formation of mRNA 5'-terminal caps, was isolated from HeLa cell nuclei. The partially purified preparation, after incubation with [alpha-32P]GTP, yielded a single radiolabeled polypeptide by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The guanylylated product was stable at neutral and alkaline pHs and had a pI of 4 by isoelectric focusing. An apparent molecular weight of approximately 68,000 was estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis under reducing conditions. The formation of a covalently linked, radiolabeled GMP-protein complex and the associated release of PPi required the presence of [alpha-32P]GTP and divalent cations and incubation between pH 7 and 9. Reaction with [beta 32P]GTP, [alpha-32P]CTP, [alpha-32P]UTP, or [alpha-32P]ATP did not label the approximately 68,000-dalton polypeptide. Phosphoamide linkage of the GMP-enzyme complex was indicated by its sensitivity to cleavage by acidic hydroxylamine or HCl and not by NaOH or alkaline phosphatase. Both formation of the GMP-enzyme intermediate and synthesis of cap structures of type GpppApG from GTP and ppApG were remarkably temperature independent; the rates of enzyme activity at 0 to 4 degrees C were 30% or more of those obtained at 37 degrees C. Radiolabeled GMP enzyme complex, isolated by heparin-Sepharose chromatography from reaction mixtures, functioned effectively as a GMP donor for cap synthesis with 5' diphosphorylated oligo- and polynucleotide acceptors. Alternatively, protein bound GMP could be transferred to PPi to form GTP. The formation of a guanylylated enzyme intermediate appears to be characteristic of viral and cellular guanylyltransferases that modify eucaryotic mRNA 5' termini. PMID- 6290878 TI - Murine T-lymphoma 'immunoglobulin' is identical to leukemia virus gp 70. AB - We have used chicken anti-mouse immunoglobulin antiserum to precipitate molecules from mouse T-lymphoma cells that had been radioiodinated. We analysed the immunoprecipitates by two-dimensional gel electrophoresis and compared the results with immunoprecipitates generated by other antisera. We found that the molecule precipitated by chicken anti-mouse immunoglobulin from T-lymphoma cells was identical to the mouse leukemia virus envelope glycoprotein (gp 70) produced by the T-lymphoma. Mouse IgM myeloma proteins block precipitation of T-lymphoma molecules by chicken anti-mouse immunoglobulin. We conclude that mouse IgM and gp 70 share antigenic determinants. This may lead to erroneous conclusions about the presence of immunoglobulin on T-cells. PMID- 6290879 TI - [Treatment of malignant tumors. Therapeutic possibilities and organizational requirements]. PMID- 6290880 TI - Polyneuropathy, papilledema, and plasmacytoma. PMID- 6290881 TI - The frequency of muscle involvement in myasthenia gravis correlates with mean muscle temperature. PMID- 6290882 TI - [Clinical features, diagnosis and therapy of chromomycosis]. PMID- 6290883 TI - Successful treatment of chronic esophageal candidosis with oral ketoconazole. PMID- 6290884 TI - In vitro susceptibility of Basidiobolus haptosporus and other zygomycetes to ketoconazole. PMID- 6290885 TI - Multiple sclerosis (second of two parts). PMID- 6290886 TI - Masking of macrocytosis by alpha-thalassemia in blacks with pernicious anemia. PMID- 6290887 TI - Progressive multifocal leukoencephalopathy in a male homosexual with T-cell immune deficiency. PMID- 6290888 TI - Peripheral neuropathy. PMID- 6290889 TI - Cutaneous phaeohyphomycosis of the sole caused by Exophiala jeanselmei and its susceptibility to amphotericin B, 5-FC and ketoconazole. PMID- 6290890 TI - Selective effects of two systemic fungicides on soil fungi. AB - BAS 317 00F was not toxic to the total count of fungi after 2 days but was regularly significantly toxic at the three doses after 5, 20 and 40 days and toxic at the low and the high doses after 80 days. In the agar medium, it was toxic to the counts of total fungi. Aspergillus, A. terreus, Rhizopus oryzae and Mucor racemosus at the high dose. Only the mycelial growth of Trichoderma viride which was significantly inhibited by the three doses when this fungicide was added to the liquid medium. Polyram-Combi induced two effects on the total population of soil fungi. One inhibitory and this was demonstrated almost regularly after 2, 10 and 40 days and the other stimulatory after 80 days of treatment with the low and the high doses. In the agar medium, this fungicide was very toxic to total fungi and to almost all fungal genera and species at the three doses. Several fungi could survive the high dose. In liquid medium, the test fungi showed variable degree of sensitivity and the most sensitive was Gliocladium roseum which was completely eradicated by the three doses. PMID- 6290891 TI - Bacterial toxins and cyclic AMP. PMID- 6290892 TI - Dihydroouabain is an antagonist of ouabain inotropic action. AB - The Na+, K+-pump controls a wide variety of cellular systems and its inhibition by cardiac glycosides modifies important physiological functions and evokes several pharmacological effects (refs 1, 2 and refs therein). However, not all the actions of cardiac glycosides can be attributed to Na+, K+-pump inhibition and several observations show that, at low doses, cardiac glycosides stimulate the pump. It has been proposed that their positive inotropic effect could be the sum of two processes: the inhibition of the pump and a still unknown additional inotropic mechanism. In guinea pig heart, low doses of ouabain interact with high affinity binding sites, which differ from the lower-affinity sites responsible for Na+, K+-pump inhibition. It has been suggested that ouabain interaction with these high-affinity sites could be responsible for the additional inotropic mechanism. The existence of two classes of ouabain-binding sites has been documented not only in guinea pig heart, but also in dog, rat and human heart. Dihydroouabain, a derivative of ouabain in which the lactone ring is saturated, is about 50-fold less potent than ouabain as an inhibitor of Na+, K+-pump and does not stimulate the pump at low doses. Its inotropic effect can be entirely accounted for by the inhibition of the pump. We have examined the pharmacological action of ouabain in the presence of dihydroouabain and report here that dihydroouabain reduces ouabain inotropic action but not Na+, K+-pump inhibition. PMID- 6290893 TI - Preferential effect of gamma interferon on the synthesis of HLA antigens and their mRNAs in human cells. AB - Interferons produce a variety of biological effects on cells. They induce resistance to virus proliferation, inhibit cell growth, modify cell structure and differentiation, stimulate some immune functions and inhibit others. However, the different interferon (IFN) species may vary in their mechanism of action and, hence, in their relative efficiency for inducing each of the effect. IFN-gamma (type II) appears to show stronger immunoregulatory and growth inhibitory effects than antiviral effects, but this conclusion has been challenged in other reports. The aim of the present work is to compare the action of IFN-gamma and other (type I) interferons on the induction of (2'-5') oligo(A) synthetase which is probably part of the antiviral response and the induction of the histocompatibility HLA-A, B,-C antigens. We have shown previously that the induction of both proteins is regulated by interferons at the mRNA level, but show here that IFN-gamma from stimulated human lymphocytes and from monkey cells transfected by cloned human IFN-gamma cDNA induced the HLA-A,-B,-C and beta 2-microglobulin mRNAs or proteins at concentrations over 100 times lower than those needed to induce the (2' 5')oligo(A) synthetase and the antiviral state. This difference was not found with IFN-alpha and -beta (type I). PMID- 6290894 TI - Genetic and molecular mechanisms of viral pathogenesis: implications for prevention and treatment. AB - The pathogenesis of infection of mice by the mammalian reoviruses involves several discrete steps. Each of the three viral outer capsid proteins has a highly distinct and specialized role: one protein (sigma 1) binds to cell surface receptors; a second protein (mu 1C) determines the capacity for viral growth at mucosal surfaces; and the third protein (sigma 3) is responsible for inhibiting cell macromolecular synthesis. A detailed picture of the molecular basis of reovirus virulence and attention is now emerging. PMID- 6290895 TI - A molecular map of the immune response region from the major histocompatibility complex of the mouse. AB - A stretch of 200 kilobases (kb) of DNA from the I region of the mouse major histocompatibility complex has been cloned and characterized. It contains the genes for the biochemically defined class II proteins E alpha, E beta and A beta. DNA blot analyses suggest that the I region may contain only 6-8 class II genes. Correlation of our molecular map with the genetic map of the I region confines two of the five I subregions, I-J and I-B, to less than 3.4 kb of DNA at the 3' end of the E beta gene where a hotspot for recombination has been observed. Indeed, the I-A and I-E subregions may be contiguous. If so, the I-B and I-J subregions are not encoded in the I region between the I-A and I-E subregions. PMID- 6290896 TI - Cholecystokinin activation of single-channel currents is mediated by internal messenger in pancreatic acinar cells. PMID- 6290897 TI - Mechanism of activation of a human oncogene. AB - The oncogene of the human EJ bladder carcinoma cell lines arose via alteration of a cellular proto-oncogene. Experiments are presented that localize the genetic lesion that led to activation of the oncogene. The lesion has no affect on levels of expression of the oncogene. Instead, it affects the structure of the oncogene encoded protein. PMID- 6290898 TI - Pathological studies on encephalitis in mice experimentally inoculated with bovine coronavirus. AB - One- to 21-day-old mice were examined pathologically after inoculated intracerebrally or subcutaneously with the Kakegawa strain of bovine coronavirus. In 1- to 4-day-old mice inoculated intracerebrally, the brain contained a small number of neutrophils and lymphocytes having infiltrated diffusely and perivascularly and some degenerative neurons. In mice inoculated intracerebrally at 7 to 21 days of age, severe necrosis of pyramidal cells was shown in Ammon's horn. Perivascular infiltrations of neutrophils and lymphocytes were moderate to severe. Some neurons were degenerative in the cerebral cortex, thalamus and midbrain. Degeneration of some neurons and mild infiltration of neutrophils and lymphocytes were observed in the brain of mice inoculated subcutaneously at 1 to 7 days of age. Perivascular infiltration of neutrophils and lymphocytes was prominent in the cerebral cortex of mice inoculated subcutaneously at 14 days of age. Cellular infiltration was also seen in the thalamus, Ammon's horn, midbrain, cerebellum and medulla oblongata. All the mice, except one, inoculated subcutaneously at 21 days of age were free from neural changes. Electron microscopically, virus particles were observed in and outside of the degenerative neurons. They had a core 70 nm in diameter and an envelope with spikes. PMID- 6290899 TI - Survey on antibody against egg drop syndrome-1976 virus among chicken flocks in Japan. AB - A survey on antibody against egg drop syndrome-1976 virus was carried out with 4,518 sera collected from 667 flocks over a period from 1971 to 1981 in Japan. Antibody-positive sera were initially detected in 1976, and the number of positive sera increased suddenly in 1979, and a total of 270 sera from 23 flocks were positive. The positive flocks consisted of 20 broiler parent flocks belonging to 9 farms and 3 layer flocks belonging to 2 farms. It seemed from the results of this survey that the prevalence of the virus might have been limited in Japan. PMID- 6290900 TI - The effect of microelectrophoretically applied clonidine on single cerebral cortical neurones in the rat. Evidence for interaction with alpha 1 adrenoceptors. AB - The technique of microelectrophoresis was used in order to examine the effects of clonidine on single neurones in the somatosensory cortex of the rat, and to compare its actions with those of noradrenaline and phenylephrine. Clonidine evoked only excitatory responses on cortical neurones. The clonidine-sensitive neurones were also excited by noradrenaline and phenylephrine. Clonidine had a consistently lower apparent potency than either noradrenaline or phenylephrine. Responses to clonidine had a slower time-course than responses to the other two adrenoceptor agonists, both the latencies to onset and the recovery times being longer for responses to clonidine than for responses to noradrenaline and phenylephrine. When the mobilities of clonidine and phenylephrine were compared using an in vitro method, no significant difference was found between the mobilities of the two ionic species, suggesting that they have similar transport numbers. Thus the difference between the potencies and time-courses of responses to clonidine and phenylephrine are presumably of biological origin. Responses to clonidine were antagonised by microelectrophoretically applied prazosin; responses to phenylephrine were equally antagonised, while responses to acetylcholine were not affected. Clonidine could reversibly antagonise excitatory responses to both noradrenaline and phenylephrine, without affecting responses to acetylcholine. The results suggest that clonidine may act as a partial agonist at excitatory alpha 1-adrenoceptors on cortical neurones. PMID- 6290901 TI - Involvement of catecholamines in Haemophilus influenzae induced decrease of beta adrenoceptor function. AB - The deeper airways of patients with asthmatic bronchitis are often infected with Haemophilus influenzae. Vaccination of guinea pigs with H. influenzae resulted in a significant impairment of the isoproterenol induced relaxation of isolated tracheal spirals by approximately 50% 4 days following vaccination. In the present study we further investigated the effects of some drugs affecting catecholamine release on the H. influenzae induced functional desensitization of tracheal spirals. Benserazide, an inhibitor of dopa-decarboxylase, completely prevented the reduction in isoproterenol-induced relaxation after H. influenzae vaccination, while no effect on relaxation of tracheal spirals from control animals was detected. On the other hand, inhibiting the re-uptake of catecholamines with desipramine did not influence the relaxation in the H. influenzae vaccinated tracheal spirals. Treatment of control animals with desipramine however resulted in a decreased relaxation of the isolated spirals by 40%. One day following vaccination with H. influenzae the level of norepinephrine in lung tissue was significantly elevated by 71%, and in plasma by 77%, while after 4 days no significant effects were observed. The spontaneous release of norepinephrine, epinephrine and dopamine of tracheal incubates was increased at days 1 and 4 following vaccination. The release of catecholamines from minced lung incubates of H. influenzae pretreated guinea pigs did not differ from that of controls. On the basis of these results it may be suggested that catecholamine metabolism is changed in lungs from H. influenzae vaccinated animals. Catecholamines, accordingly may play a role in the desensitization of beta adrenoceptors by H. influenzae. PMID- 6290902 TI - beta-Adrenoceptor ligand binding and supersensitivity to isoprenaline of ventricular muscle after chronic reserpine pretreatment. AB - 1. Isolated papillary muscles from guinea-pig hearts were paced at a constant frequency and isometric contractions reported. 2. Guinea-pigs were either untreated or pretreated with reserpine. Three pretreatment schedules were used; a) 0.5 mg kg-1 i.p. at 24 h, b) 5.0 mg kg-1 at 72 h and 3.0 mg kg-1 at 48 and 24 h, or c) 0.1 mg kg-1 daily for 7 days. 3. Cumulative concentration-response curves for the isoprenaline-induced increases in tension were obtained. The geometric mean EC50 values after the 3 and 7 day reserpine pretreatment schedules were significantly (P less than 0.05) less than for untreated guinea-pigs indicating a supersensitivity. 4. EC50 values for the positive inotropic responses to histamine and calcium in papillary muscles from reserpine-pretreated guinea-pigs did not differ significantly (P less than 0.05) from those from untreated animals. This suggests that the supersensitivity to isoprenaline is beta-adrenoceptor specific. 5. Membrane fractions were prepared from the ventricles of the untreated and reserpine-pretreated guinea-pigs from which papillary muscles had been removed. Binding of [3H]-dihydroalprenolol ([3H]-DHA) to beta-adrenoceptors of these membranes was determined. Equilibrium dissociation constants (KD) and total numbers of binding sites (Bmax) were determined by Scatchard analysis of the saturation curves for [3H]-DHA binding. 6. There was no increase in affinity (fall in KD value) or change in the total number of binding sites associated with reserpine-induced supersensitivity. The equilibrium inhibition constant (Ki) for the displacement of [3H]-DHA binding by isoprenaline was also identical in membranes from untreated and reserpine-pretreated animals. Thus reserpine-induced supersensitivity to isoprenaline does not appear to involve a change in affinity for the beta-adrenoceptor or in receptor numbers as determined by [3H]-DHA binding. PMID- 6290903 TI - Seasonal variation in responses of the toad renal vasculature to adrenaline. AB - Adrenaline and noradrenaline produced both constriction and dilatation of the toad renal vasculature: constrictor effects were mediated by alpha-adrenoceptors; dilator effects were mediated by beta-adrenoceptors. Vasoconstriction was the predominant response to these amines. Dilatation was only revealed after blockade of alpha-adrenoceptors and constriction of the vasculature. There was a marked seasonal variation in the constrictor responses to adrenaline, but not to noradrenaline. The maximal increase in renal vascular resistance produced by adrenaline in summer was greater, by a factor of three, than the maximum constrictor response in winter. The response to adrenaline in summer was also significantly greater than the responses to noradrenaline in both summer and winter. However, after treatment with propranolol there was no difference between the maximum vasoconstrictions to these two amines, in summer or in winter. Determination of dissociation constants for phentolamine indicated that adrenaline and noradrenaline acted on the same population of alpha-adrenoceptors in both summer and winter. The enhanced vasoconstriction to adrenaline in summer appears to be due to a reduced potency of adrenaline on renal vascular beta adrenoceptors. The reduction in potency may be caused by a subtle configurational change in the beta-adrenoceptors, perhaps induced by hormonal changes associated with the onset of breeding. PMID- 6290904 TI - Noradrenaline inhibits central serotonin release through alpha 2-adrenoceptors located on serotonergic nerve terminals. AB - The effect of noradrenaline on the depolarization-evoked release of 3H-5 hydroxytryptamine was investigated in superfused synaptosomes prepared from rat cortex and hippocampus and prelabelled with the radioactive indoleamine. Noradrenaline reduced in a concentration-dependent way the release of 3H-5 hydroxytryptamine elicited by 15 mM KCl. The inhibition was counteracted by the alpha-adrenoceptor antagonists phentolamine or yohimbine, but not by prazosin. The results indicate that, in rat brain, the inhibition of 5-hydroxytryptamine release by noradrenaline is mediated by adrenoceptors of the alpha 2-type localized on the terminal serotonergic fibres. PMID- 6290906 TI - [Neuropathy of the ulnar nerve in the elbow region and its treatment]. PMID- 6290905 TI - [Cholesterol metabolism in atherosclerosis. I. Receptor specific catabolism]. PMID- 6290907 TI - [Insulinoma]. PMID- 6290908 TI - [Percutaneous transhepatic portography for the localization of insulinoma]. PMID- 6290909 TI - [Radiodiagnosis of insulinomas]. PMID- 6290910 TI - Interactions between the growth effects of a kidney epithelial cell growth inhibitor and extracellular concentrations of cyclic nucleotides. AB - The effect of dibutyryl cyclic nucleotides, dbcAMP and dbcGMP, on the growth of BSC-1 cells has been studied in the presence and absence of kidney epithelial cell growth inhibitor. The growth response of the cells was dependent on concentrations of the dibutyryl cyclic nucleotides and on the presence or absence of serum in the medium. In the presence of serum, a high concentration (10(-3) M) of dbcGMP largely overcame the action of the kidney epithelial cell growth inhibitor. In the absence of serum, a high concentration (10(-3) M) of dbcAMP increased growth inhibition observed with the growth inhibitor. In the presence of serum, low concentrations of dbcAMP were growth stimulatory and partially overcame the action of the growth inhibitor. PMID- 6290911 TI - Epstein-Barr virus is not associated with head and neck neoplasms other than nasopharyngeal carcinoma. PMID- 6290912 TI - Cell surface alterations of avian sarcoma virus B77- and 3,4-benzo(a)pyrene transformed rat fibroblasts. I. Cell surface proteins characterized by two dimensional electrophoresis. AB - Plasma membrane proteins of avian sarcoma virus B77-, 3,4-benzo(a)pyrene- and spontaneously transformed rat fibroblasts were metabolically- and cell surface radiolabeled and analyzed by electrophoresis under denaturing conditions (SDS PAGE) or by two-dimensional electrophoresis. Most extensive series of protein alterations on transformed cells was detected by two-dimensional electrophoresis of cell surface proteins radiolabeled by lactoperoxidase catalyzed radioiodination: a fibronectin-like 220k protein with two isoelectric forms (pI 5.9 and 6.2-6.6), a 180k protein of pI 6.2-6.6 and two proteins of 48k and 50k with pI approximately 5.4-all decreased on examined transformed cells. In addition to these proteins, a series of proteins more markedly detectable on transformed cells was found: a 110-120k (pI 5.6) protein found particularly on 3,4-benzo(a)pyrene- and spontaneously transformed cells, a series of proteins in the region of molecular weights 46-54k with pI 5.5-6.0, two proteins of 35k (pI 6.2 and 6.4) and two further, basic proteins of 38k (pI approximately 7.2 and 7.8). Some of these changes were visualized also by further techniques for radiolabeling of proteins, i.e. reductive methylation of cell surface proteins, metaboling radiolabeling by 3H-leucine followed by plasma membrane isolation, as well as by protein staining after two-dimensional electrophoresis of isolated plasma membrane proteins. A 70k and a 120k radioiodinated proteins were immunoprecipitated from all transformed cells and, to a markedly lesser extent from untransformed fibroblasts by the immune serum against rat type-C endogenous virus. A similar 70k protein was immunoprecipitated from avian sarcoma virus B77 transformed cells also by the antiserum against Friend murine leukemia virus envelope glycoprotein gp70. PMID- 6290913 TI - Cell surface alterations of avian sarcoma virus B77-and 3,4-benzo(a)pyrene transformed rat fibroblasts. II. Cell surface glycoproteins characterized by two dimensional electrophoresis. AB - Cell surface proteins of avian sarcoma virus B77-, 3,4-benzo(a)pyrene- and spontaneously transformed rat fibroblasts were tritium-radiolabeled by mild periodate oxidation followed by tritiated sodium borohydride reduction and by galactoseoxidase/NaB3H4 technique. Radiolabeled cell surface sialoglycoproteins were analyzed by electrophoresis under denaturing conditions (SDS-PAGE), or by two-dimensional electrophoresis (isoelectric focusing, SDS-PAGE). Following major glycoproteins were quantitatively decreased on all examined transformed cells: a 220k glycoprotein with pI of approximately 6.2, a 180k glycoprotein with pI of approximately 6.0-6.2, a 110k glycoprotein (pI 6.2). A series of further sialoglycoproteins was quantitatively increased on all examined transformed cells, as follows: a markedly increased 70k glycoprotein (pI approximately 4.8) and a 120k glycoprotein (pI 5.0-5.2) increased more markedly on onco-virus transformed and spontaneously transformed cells. Further sialogalactoprotein (28k, pI 5.8) was visualized as increased on all examined transformed cells only by galactoseoxidase/NaB3H4 technique. A 70k sialoglycoprotein was immunoprecipitated from transformed cells by the immune serum against Friend murine leukemia virus envelop glycoprotein gp70 and, to a markedly lesser extent, from untransformed cells. A similar glycoprotein, together with an another 120k glycoprotein were precipitated from transformed cells and to a minor extent from untransformed cells also by an antiserum against endogenous rat type-C virus. PMID- 6290914 TI - Studies on antibodies reactive with glycoproteins of primate type C viruses in patients with myeloid leukemias and with potential preleukemia. AB - Human blood plasma from patients with myeloid leukemias, potentially preleukemic disorders and healthy individuals contains antibodies which react with purified glycoproteins of the baboon endogenous virus and the gibbon ape leukemia virus. Experiments are presented which illustrate characteristic distributions of antiviral antibodies in the plasma of different investigated groups. The presence of high-titer antibodies is associated with remission of acute myeloid leukemia and longer survival of patients with preleukemia. PMID- 6290915 TI - [The effect of lithium on calcium-dependent processes in nerve cells. Attempt at an explanation for therapeutic observations based on membrane physiologic examinations]. PMID- 6290916 TI - [Porphyria cutanea tarda, polyneuropathy and myopathy in small cell bronchial cancer]. PMID- 6290917 TI - [Treatment of tumors in relation to the transverse portion of the vertebral artery]. AB - Among 28 lesions, 22 tumors and 6 anteriovenous malformations, operated with control of the vertebral artery, 15 are located close to the transverse canal. Analysis of arteriographic datas from 20 cases of tumors related to the vertebral artery, points out the interest of the surgical control of this vessel, in the treatment of these tumors. This control is mainly indicated in case of hypervascularized tumors, or compression of the vertebral artery, especially if it is the dominant vessel or if it gives a radiculo-medullary branch. For this control, the lateral retro-jugular approach is used, and sometimes associated with other techniques: embolization, posterior approach, or anastomosis between carotid and vertebral artery about the lesion. PMID- 6290918 TI - Changes in neurotransmitter actions in the aged rat hippocampus. AB - Intracellular activity was recorded from pyramidal cells in region CA1 of young and aged rat hippocampal slices. There were no apparent differences between the two age groups in the passive or active membrane properties tested. These included resting membrane potential, input resistance, spike size and overshoot, after potentials, and EPSP's. There were marked differences between the two age groups in reactivity to chemical stimulation. The response to acetylcholine (ACh) in young rat hippocampal cells consisted of an initial hyperpolarization followed by a late, slow depolarization. These were accompanied by a decrease in reactivity of the recorded cell to afferent stimulation. In the old cells the initial hyperpolarization and late depolarization were markedly reduced, yet the decreased reactivity to afferent stimulation was maintained. Both young and old cells were depolarized by GABA to the same magnitude, yet the recovery was slower in the old cells. 5-HT evoked a smaller response in old compared to young cells. These data indicate that some specific physiological processes which are not paralleled by reported biochemical changes are affected by aging. The mechanisms underlying these changes are subject to further investigations. PMID- 6290920 TI - [Steroid treatment in neurosurgical practice -the effects of glucocorticoid on adrenal cortical function]. PMID- 6290919 TI - [Treatment of salaam seizures in children at the Maternal and Child Care Specialist Center in Cracow]. PMID- 6290921 TI - Secretion of corticotrophin and its hypothalamic releasing factor in response to morphine and opioid peptides. AB - The influence of morphine and enkephalins on the functional activity of the hypothalamo-pituitary-adrenocortical system in the rat was studied by investigating their effects on the secretion in vivo and in vitro of corticotrophin (ACTH) by the pituitary gland and corticotrophin-releasing factor (CRF) by the hypothalamus. A single injection of morphine caused a rise followed by a fall in hypothalamic CRF content and increases in the concentrations of ACTH in the plasma and adenohypophysis. In addition, the stress-induced increments in hypothalamic CRF and pituitary and plasma ACTH were exaggerated in morphine treated rats. The production of ACTH by pituitary segments in vitro was not affected by the addition to the incubation medium of morphine, met-enkephalin, leu-enkephalin or naloxone. However, morphine and the enkephalins stimulated the secretion of CRF by isolated hypothalami and their effects were antagonized by naloxone. The results indicate that morphine and the enkephalins evoke hypothalamo-pituitary-adrenocortical activity by stimulating specific receptors in the hypothalamus and raise the possibility that opioid peptides and their receptors are physiologically important in the control of the secretion of CRF. PMID- 6290922 TI - Differential effects of catechol estrogens, progestins and CI-628 administered by constant infusion on the central and peripheral action of estradiol. AB - By means of continuous infusion by Alzet minipumps, we tested the ability of the catechol estrogens, 2-hydroxyestrone (2-OHE1) and 2-hydroxyestradiol (2-OHE2), as well as two progestins and the anti-estrogen, CI-628, to block the action of estradiol (E2) on sexual behavior, LH surge frequency, uterine weight increase and glucose-6-phosphate dehydrogenase (G6PDH) activity in the uterus and pituitary of ovariectomized rats. Neither catechol estrogen produced inhibitory effects, whereas the potent progestin, R-5020 (5 micrograms/h), and CI-628 (50 micrograms/h) inhibited the E2-induced increase in the LH surge frequency and lordosis response. Progesterone at 20 micrograms/h and CI-628 at 5 micrograms/h did not influence these end points. In the pituitary and uterus, the concurrent administration of R-5020 and E2 attenuated the increase of G6PDH activity. As expected, CI-628 inhibited the action of E2 on G6PDH but was more effective in doing so at the lower of the two doses. CI-628 (50 micrograms/h) also blocked the neuroendocrine and behavioral effects of 2-OHE2 (1 microgram/h) and 4 hydroxyestradiol (0.1 microgram/h). Taken together with our previous studies, these results are consistent with the notion that the catechol estrogens exert effects on sexual behavior and LH surge frequency by virtue of their estrogenic properties rather than by other means. PMID- 6290923 TI - Immunocytochemical localization of angiotensin immunoreactivity in gonadotropes and lactotropes of the rat anterior pituitary gland. AB - Utilizing an immunocytochemical technique wherein two different antigens can be visualized on the same tissue section, angiotensin-like immunoreactivity was found in rat adenohypophyseal cells that showed staining with antisera to hLH, hFSH or human prolactin. Angiotensin-like immunoreactivity was not found in cells that showed staining with antisera to hTSH, C-terminal ACTH or hGH. The nature and possible physiologic significance of the angiotensin-like immunoreactivity is discussed. PMID- 6290924 TI - Release of prolactin in response to microinjection of morphine into mesencephalic dorsal raphe nucleus. AB - Blood samples were collected via jugular catheters from ovariectomized rats at 10 min intervals for 1 h before and 2 h after microinjection of 0.5 microliter of either saline vehicle or morphine sulfate (10 micrograms) into the dorsal raphe nucleus (DRN) or adjacent periaqueductal gray by means of chronically-implanted guide cannulae. Prolactin was measured by radioimmunoassay, and mean preinjection and mean postinjection values were compared for each rat (t test) as well as for each treatment group (paired t test). Neither saline in DRN nor morphine in the surrounding periaqueductal gray significantly altered circulating prolactin. A significant rise in prolactin was observed following injection of morphine into DRN. This effect of morphine was prevented by pretreatment of the animals with the narcotic antagonist naltrexone (10 mg/kg i.v.), indicating the involvement of opiate receptors. These results indicate that DRN is one site at which systemically-administered morphine might act, and suggest the possibility of participation of this mechanism in modulation of prolactin release by endogenous opioids. PMID- 6290925 TI - Biochemical and physiological studies of the beta-adrenoceptor and the D-2 dopamine receptor in the intermediate lobe of the rat pituitary gland: a review. AB - The intermediate lobe (IL) of the rat pituitary gland responds to catecholamines. Catecholamines interacting with the beta-adrenoceptor stimulate adenylate cyclase activity, enhance cyclic AMP formation and thereby trigger the release of alpha melanocyte-stimulating hormone (alpha-MSH). Catecholamines interacting with a D-2 dopamine receptor (in the classification schema of Kebabian and Calne) diminish adenylate cyclase activity and thereby decrease the capacity of IL cells to synthesize cyclic AMP. Dopaminergic agonists also inhibit the release of alpha MSH from IL cells. The homogeneity of the IL facilitates biochemical investigations of this tissue. PMID- 6290926 TI - Use of ACTH fragments of children with infantile spasms. PMID- 6290927 TI - Trial of an ACTH4-9 Analogue (ORG 2766) in children with intractable seizures. AB - The ACTH4-9 Analogue (ORG 2766) is an orally administered neuropeptide which has been showed to have behavioral effects in human subjects, but does not have any steroidogenic effects, nor other significant side effects. This study was a double-blind, crossover trial of the ACTH4-9 analogue in four children with intractable seizures. There was some slight improvement in seizure frequency in two patients, with an equivocal improvement in a third child. A fourth child was unchanged, while the fifth dropped out of the study. There were no side effects from the therapy. This questionable improvement in seizure frequency in some of the most intractable patients with seizure disorders indicates the need to further assess this ACTH4-9 analogue in a larger study over a longer period of time. PMID- 6290928 TI - Neural and vascular tissue reaction of cyanoacrylate adhesives: a further report. AB - The effect of a new tissue adhesive, carbohexoxymethyl 2-cyanoacrylate monomer (Ethicon CHC), was evaluated in six cats divided into two groups. With the cats under barbiturate anesthesia. Ethicon CHC was applied to the left cruciate cortex and the left femoral neurovascular bundle. Normal saline was applied to the right side for control. The first group (n = 4) and the second group (n = 2) were killed 4 and 7 days, respectively, after application of the adhesive. Neuropathological examination revealed meningeal astrocytosis, vascular wall degeneration, hemorrhage, and inflammatory reaction in both groups. This adhesive does not display the ideal property of inertness, which would permit its safe use. Focal tissue reactions caused by the adhesives in sensitive areas of the cortex can result in significant neurological deficit. PMID- 6290929 TI - Transplantable canine glioma model for use in experimental neuro-oncology. AB - The need for a large animal tumor model in experimental neuro-oncology led us to re-evaluate and to modify the transplantable canine glioma of Wodinsky and Walker. Successive passages of the original tumor brei were made in purebred beagles, from beagle to mongrel, and between various mongrel strains until an intracerebral injection of 0.1 cc on Days 1 to 3 of life produced a 93% incidence of tumor take in all breeds. The mean survival was 13.5 +/- 1.9 days after injection (range, 10 to 19 days) in 10 litters. The tumor was invariably fatal and possessed many of the histological characteristics of human glioblastoma (i.e., capillary proliferation, pseudopallisading, frequent mitotic figures, and multinucleated giant cells). The animals were large enough to be scanned on the Pfizer 450 scanner, and the tumors were visualized in vivo as typical "ring" lesions after the injection of contrast agent. Intravital staining with Evans blue outlined the areas of contrast enhancement observed in the same tumors by computed tomography. The apparent defect in the blood-brain barrier could be explained in part by the absence of endothelial tight junctions on electron microscopy. Stability in the histology and activity of the tumor could be demonstrated after more than 14 months of storage at -70 degrees C. The transplantable canine glioma model has many advantages including low cost, reproducible morphology, a short survival time, and relative safety for the investigator. The large size of the animal preparation allows the use of complex surgical instrumentation and radiographic study, as well as repeated sampling of cerebrospinal and other fluids. PMID- 6290930 TI - Clinical toxicity of combined modality treatment with nitrosourea derivatives for central nervous system tumors. AB - Two nitrosourea compounds--1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU) and 1-(2 chloroethyl)-3-cyclohexyl-1-nitrosourea (CCNU)--have been used in the treatment of primary and metastatic brain tumors after operation and/or radiotherapy. Hematological and nonhematological toxicity were recorded in 272 patients treated between May 1973 and June 1978. BCNU was given to 135 patients (80 mg/m2 i.v. daily for 3 days) and CCNU was given to 137 patients (130 mg/m2 orally, single dose) every 8 weeks until progression of the primary disease process or for a total of 12 cycles. Radiation therapy (5500 +/- 500 rads in 6 to 7 weeks) was carried out after the first course of chemotherapy. BCNU and CCNU induced the same hematological and clinical toxicity. The bone marrow toxicity seemed to be dose-related, delayed, and cumulative. One case of acute nonlymphoblastic leukemia arising 2 months after the end of CCNU therapy is reported. PMID- 6290931 TI - Origin and properties of spinal cord field potentials. AB - Electrical or natural stimulation of cutaneous, muscle, or visceral nerves evokes a field potential in the spinal cord; the field potential elicited by each type of afferent fiber differs in wave form. Cutaneous nerve-evoked spinal cord field potentials consist of a triphasic spike, one to three negative waves, and a slow positive wave. Muscle nerve-evoked spinal cord field potentials are composed of a triphasic spike, a slow positive wave, and one to three negative waves with different relative latencies and durations than the negative waves evoked by cutaneous nerves. Visceral nerve-evoked potentials in the spinal cord comprise only a single negative wave and a slow positive wave. The triphasic spike is a compound action potential propagating through the large primary afferent fibers; the negative waves are generated by dorsal horn interneurons; the positive wave is a reflection of primary afferent depolarization. A knowledge of the origin and properties of spinal cord field potentials has practical usefulness for both the experimental neurophysiologist and the clinician. PMID- 6290932 TI - Simple reaction times to lateralized visual stimuli in a case of callosal agenesis. AB - Simple reaction times (RTs) of a young acallosal woman were measured using three different intensity levels of the visual stimulus. It was found that "'crossed" stimulus/hand combinations yielded finger RTs about 20 msec longer, on average, than "uncrossed" combinations. In addition, an inverse relation was found between stimulus intensity and the magnitude of this crossed/uncrossed difference in RT. It was also found that vocal RTs displayed a large advantage (18 msec) for right visual-field stimulation. It is suggested that the stimulus information in crossed finger reactions must cross the midline through visually-coded neurones in the acallosal brain. PMID- 6290933 TI - Changes in acetylcholine sensitivity of sensomotor cortical neurons during extinction of a conditioned reflex. PMID- 6290934 TI - Change in the receptive fields of the visual cortex of the cat in relation to the level of wakefulness. PMID- 6290935 TI - Characteristics of unit activity in hippocampal structures (the subiculum). PMID- 6290936 TI - Dynamics of the response of neurons of the visual cortex of the rabbit to repetitive nonvisual stimuli and their complexes with light. PMID- 6290937 TI - Suppressing effect of hydrocortisone and corticotropin on the self-stimulation reaction. PMID- 6290938 TI - Anterograde tracer and field potential analysis of the neocortical layer I projection from nucleus ventralis medialis of the thalamus in cat. AB - The projection of the ventromedial nucleus of the thalamus to the neocortex was studied in cat by means of anterograde and retrograde transport of horseradish peroxidase, by the depth profile of evoked thalamocortical field potentials, and by superfusion of the cortex with manganese to block transmitter release. Horseradish peroxidase injected into the ventromedial nucleus was anterogradely transported to the outer third of layer I in the neocortex, extending from the depth of the cruciate sulcus anterior to the olfactory bulb and tract. The region of projection from the ventromedial nucleus extended mediolaterally from the medial wall of the proreus gyrus to the ventral tip of the coronal gyrus. Horseradish peroxidase injections or applications in these areas of the neocortex resulted in the retrograde labeling of neurons in the ventromedial nucleus. Injections in many other cortical areas did not result in labeled neurons in this nucleus. Stimulation of the ventromedial nucleus with single pulses elicited surface-negative waves in the medial part of the precruciate region that had superficial isoelectric points. Superfusion of the precruciate area with manganese resulted in the suppression of the ventromedial-evoked wave, whereas control extracellular waves in deeper layers were unaffected. An additional additional finding was that horseradish peroxidase injections in the ventromedial nucleus led to a dense reciprocal retrograde labeling of neurons in layer VI of that part of the cortex to which the ventromedial nucleus projects. We conclude that, in cat, (1) the ventromedial nucleus projects to layer I of the cerebral cortex anterior to the cruciate sulcus and receives a dense reciprocal projection from layer VI; (2) stimulation of neurons in the ventromedial nucleus causes depolarization of structures in layer I and these neurons are responsible for recruiting responses in the anterior cortex. PMID- 6290939 TI - A quantitative electron microscopic study of synaptic reorganization in the rat medial habenular nucleus after transection of the stria medullaris. PMID- 6290940 TI - LHRH pathways in rat brain: 'deafferentation' spares a sub-chiasmatic LHRH projection to the median eminence. AB - Localization of luteotrophic hormone-releasing hormone (LHRH) was examined by immunocytochemistry in untreated male rats and rats that received an anterior hypothalamic deafferentation. Gonadotrophic function was assessed by examining testicular weight and morphology. All of the antisera used in this study were able to reveal LHRH cell bodies and fibers. Cells from the medial preoptic area, (particularly the preoptic periventricular or median preoptic nuclei) and lateral anterior hypothalamus sent axons to the median eminence. The fiber tracts were loosely organized into (1) a tract that coursed through the organum vasculum of the lamina terminalis between the optic nerves and along the ventral surface of the optic chiasm; (2) a tract that coursed to the organum vasculum from the preoptic area and then traversed along the floor of the third ventricle; (3) a tract that coursed from the medial preoptic area to the median eminence along the lateral walls of the third ventricle; and (4) a tract that arose from the more caudal portions of the LHRH cell field in the lateral preoptic and lateral anterior hypothalamus, coursed along with the fibers of the medial forebrain bundle and turned medially at the caudal hypothalamus to enter the median eminence. An anterior cut which served most of the connections between the medial preoptic area and hypothalamus but did not penetrate through the optic chiasm served tract 3 and most of tracts 2 and 4, but spared the subchiasmatic projection (tract 1). The fibers that remained in the median eminence were sufficient to retain gonadotrophic function. This study provides an explanation for the variable effects of deafferentation and lesions of the anterior hypothalamus and preoptic area on gonadotrophic function. PMID- 6290941 TI - [Phosphocreatine, skeletal musculature and neuromuscular transmission. In-vitro and in-vivo research]. PMID- 6290942 TI - [Male breast carcinoma. Review of the literature and report of cases]. PMID- 6290943 TI - [Correlations between clinical aspects, electroencephalography and computed tomography in the Lennox-Gastaut syndrome]. PMID- 6290944 TI - [The glucagonoma syndrome. A clinical case]. PMID- 6290945 TI - [Use of "active" vitamin D in uremic osteodystrophy]. PMID- 6290946 TI - [Laboratory contribution in the etiologic definition of viral pericarditis]. PMID- 6290947 TI - [Malignant tumors associated with ovarian carcinoma. Study of 13 cases]. PMID- 6290948 TI - Aggregation of acetylcholine receptors in nerve-muscle cocultures is decreased by inhibitors of collagen production. AB - Coculturing of rat embryonic muscle cells with spinal cord explants resulted in the formation of large numbers of acetylcholine receptor aggregates on the myotube surface, compared to those found on muscle cells grown in the absence of nervous tissue. Remarkably fewer receptor aggregates were formed when, upon addition of nerve explants, these cocultures were treated with either cis hydroxyproline (a specific inhibitor of collagen production) or collagenase. The possibility is raised that collagen participates in the aggregation of acetylcholine receptors and in synapse formation. PMID- 6290949 TI - A transient outward current in rat sympathetic neurones. AB - Voltage-clamped rat sympathetic neurones exhibited a transient outward current similar to that previously described in invertebrate neurones and marine egg cells. It had an activation threshold of about -60 mV and displayed both voltage and time-dependent inactivation. In current clamp, this current manifested as a pronounced outward rectification at the onset of electrotonic potentials, increasing the latency to directly-evoked action potentials. The amplitude of the current was dependent on the external K+ concentration and was reduced by 4 aminopyridine. PMID- 6290950 TI - Involvement of alpha-adrenoceptors in norepinephrine-induced prostaglandin E2 release by rat pineal gland in vitro. AB - Rat pineal glands incubated in vitro with 10 or 100 microM norepinephrine (NE) released 51% and 415% more prostaglandin E2 (PGE2) to the medium than in the absence of NE. Phentolamine (10 microM) prevented fully the effect of NE at both concentrations, whereas propranolol failed to affect it significantly. After superior cervical ganglionectomy (SCGx), NE-induced PGE2 release was significantly higher than in sham-operated controls--an effect also blocked by phentolamine but not by propranolol. These results suggest that NE releases PGE2 in rat pineals via alpha-adrenoceptors, acting at a post-synaptic site, and that SCGx induces alpha-adrenergic supersensitivity in the pinealocytes. PMID- 6290951 TI - Dose-dependent interaction of the pyrethroid isomers with sodium channels of squid axon membranes. PMID- 6290952 TI - Effect of acute organophosphorus exposure on 2', 3',-cyclic nucleotide 3' phosphohydrolase activity in hen neural tissue. AB - The effect of various organophosphorus (OP) compounds on 2', 3'-cyclic nucleotide 3'-phosphohydrolase (CNP) activity and the relationship to neurotoxic response was studied. Adult White Leghorn hens were dosed with either diisopropyl fluorophosphate (DFP), tri-o-cresyl phosphate (TOCP), leptophos, or paraoxon. CNP activity was determined utilizing crude homogenates or purified myelin preparations from whole brain, spinal cord, and sciatic nerves at maximal neurologic dysfunction, evaluated as locomotor impairment. Enzyme assays of CNP revealed a decrease in brain CNP activity at the time of maximal locomotor impairment after a single, oral dose of leptophos. Decreased CNP activity was also noted in spinal cord preparations from DFP-treated hens at the time of maximal locomotor impairment. Sciatic nerve CNP activity was not altered following treatment with OP-compounds. Paraoxon, a non-neurotoxic OP-compound, had little effect on neural tissue CNP activity. Neurotoxic OP-compounds, that cause secondary degeneration of myelin (Wallerian), may be associated with decreased brain and spinal cord CNP activity at the time of maximal locomotor impairment. PMID- 6290953 TI - Effect of cytomegalovirus on renal transplantation. PMID- 6290954 TI - Addison's disease, adrenal autoantibodies and computerised adrenal tomography. AB - The present study describes 53 patients with Addison's disease, who attended Auckland hospitals between 1971-1980, 32 of whom presented for the first time during this period, when the mean annual incidence of Addison's disease was 4.5 cases/million population, and the respective frequency of idiopathic (auto immune) and tuberculous aetiologies was 92 percent and 4 percent for caucasians, but 25 percent and 63 percent for Polynesians. In addition, adrenal reserve was tested by ACTH stimulation in 20 clinically non-Addisonian patients in whom circulating adrenal antibodies had been incidentally demonstrated and present for periods of up to six years, and was normal in all cases. Such antibodies therefore lack functional disease specificity. The diagnostic value of CT adrenal scanning in illustrating the contrasting appearances between tuberculous and auto immune adrenalitis is shown in four patients. PMID- 6290955 TI - [Medroxyprogesterone acetate as glucocorticoid in combination with aminoglutethimide in the treatment of metastatic breast cancer]. AB - One of the central effects of MAP is the intrinsic glucocorticoid activity. In the therapy of metastatic breast cancer with high dose MAP the cortisol like effect could be shown even in long term treatment. The cortisol like activity of MAP leads via the suppression of ACTH to a decrease of endogenous cortisol secretion. This cortisol like activity of MAP is sufficient to replace the obligate cortisol substitution in the therapy of metastatic breast cancer with aminoglutethimide. Thus within the therapy of metastatic breast cancer the combination of two endocrine acting drugs is possible. PMID- 6290956 TI - Ectopic ACTH syndrome in APUD tumors. AB - Rapidly progressing hypercorticism was observed in 2 patients with malignant neoplasms, a 72-year-old man with anaplastic small cell carcinoma and a 48-year old woman with a primary apudoma of the liver. Both patients were treated with chemotherapy in conventional doses. Both patients developed acute dyspnea and sudden death, 7 and 10 days, respectively, after chemotherapy was given. The possibility that patients with ectopic adrenocorticotropic hormone (ACTH) secretion are excessively vulnerable to chemotherapy is raised and judicious management urged. PMID- 6290957 TI - Analysis of genetically defined liver enzymes in transplantable hepatomas and developing liver of the mouse. AB - The activities and isozyme patterns of seven enzymes involved in glycolysis, the malate-aspartate shuttle and the pentose phosphate pathway have been examined in five transplantable murine hepatomas, adult liver and fetal liver. The liver marker enzyme, alcohol dehydrogenase, has also been studied. The developmental expression of the activity and isozyme pattern of these enzymes have been determined. Seven defined loci on six different chromosomes code for these enzymes and isozymes. The tumors express no unique gene product for any of the enzymes examined, but the activities of all the enzymes in the hepatomas are more like fetal liver than adult liver, regardless of whether activities are increased or decreased in hepatomas. The activity of one enzyme is unchanged in hepatomas, and this enzyme has the same activity in fetal and adult liver. The enzyme activities in the five hepatomas vary more than the enzyme activities in liver and fetal liver of the three inbred strains of mice examined. PMID- 6290958 TI - Uracil synthesis via HCN oligomerization. AB - Uracil is released from HCN oligomers upon acid hydrolysis in concentrations of 0.001% for 1 M HCN solutions to 0.005% for 0.1 M solutions. This yield is comparable with earlier reported, minor or nonbiological pyrimidines such as 5 hydroxyuracil and orotic acid. This is the first report of uracil itself via HCN oligomerization. Data are presented which establish that the observed uracil is not formed by decarboxylation of previously formed orotic acid, but via acid hydrolysis of at least two other precursors. PMID- 6290959 TI - The radiolysis and racemization of leucine on proton irradiation. AB - D- and L-Leucine have been subjected to 39-55 percent radiolysis using 0-11 MeV protons, both with the proton beam passing through the sample or being absorbed by it, and with quenching the sample immediately on completion of irradiation or after a 21-day interval. Racemization was small (1.1-1.7 percent) and comparable in all cases, suggesting that radioracemization and secondary degradative effects were not important factors in our recent unsuccessful attempts to induce optical activity in DL-leucine by partial radiolysis using 0-11 MeV longitudinally polarized protons. PMID- 6290960 TI - Foreign body granulomas in the head and neck. PMID- 6290962 TI - Electrophysiological characteristics of dorsal horn cells in rats with cutaneous inflammation resulting from chronic arthritis. AB - (1) Electrophysiological properties of dorsal horn neurones have been investigated in decerebrate, immobilized spinal rats rendered polyarthritic by intradermal injection of Freund's adjuvant. Since arthritis is associated with pronounced erythema and oedema of the foot sole and ankle areas, this particular study was devoted to the induced modifications of responses of units driven by cutaneous inputs. It allowed comparison with previous studies performed in healthy animals. (2) Superficial dorsal horn cells could be separated in: those driven from non-oedematous skin and which had properties similar to those observed in healthy animals; and those driven from oedematous skin and which characterized by unclassical electrophysiological properties such as: (a) a relatively high level of background activity, frequently with bursting pattern and sometimes exhibiting dramatic increases; these neurones are normally silent in healthy animals; (b) a high degree of responsivity to light mechanical stimuli associated with a clear decrease in threshold. It is possible that units which initially only responded to high intensity or frankly noxious stimuli shifted to become highly or maximally responsive to gentle stimuli. (3) Similar modifications were also observed for some of the neurones located in deeper areas of the spinal cord where it is suggested that some units initially responding to both non-noxious and noxious stimuli (class 2 cells) in healthy rats shifted their characteristics to resemble units maximally driven by non-noxious stimuli (class 1 cells) in arthritic animals. (4) The numerous structural alterations reported in cutaneous tissues compared to the relatively normal structure of peripheral nerves and central nervous systems suggest that these observations can be related to peripheral sensitization of the sensory endings of thin peripheral fibres induced by the release of chemalgogens in inflammatory exudates. (5) For both superficial and deeper dorsal horn neurones, arthritis also affected the late responses of neurones to transcutaneous electrical stimulation and their responses to radiant heat. Responses to radiant heat showed an increase in threshold, sometimes associated with a very high degree of adaptation. (6) Another group of neurones displayed fading responses to repetitive mechanical stimuli. All were located in the deeper two-thirds of the dorsal horn. (7) These data indicate that chronic pathological conditions related to articular and/or cutaneous diseases strongly modify the responses of dorsal horn neurones, some being at the origin of ascending tracts. This model seems to be a promising approach to study the fundamental mechanisms involved in chronic pain in order to approach these problems in human beings. PMID- 6290961 TI - Evaluation of bone scan by scintigraphy to detect subclinical invasion of the mandible by squamous cell carcinoma of the oral cavity. AB - A prospective study using scintigraphy was performed to compare the sensitivity of the Panorex roentgenogram and the bone scan in detecting subclinical invasion of the mandible by squamous cell carcinoma of the oral cavity and floor of the mouth. Twenty-five patients with squamous cell carcinoma of the floor of the mouth were evaluated preoperatively by both the Panorex and scintigraphic techniques and the results compared with postoperative pathologic findings. In 13 (52%) of the cases, Panorex and scintigraphic techniques were comparable in detecting tumor involvement in bone. In eight cases (32%) all three modalities had normal pathologic indications of the mandible. However, in four cases (16%) results of the scintigraphic techniques were abnormal and the Panorex, normal. In four separate cases, the extent of lesion demonstrable by scintiscanning was greater than delineated by Panorex; surgical specimen confirmed this finding. Pathologic examination of operative specimens confirmed tumor involvement. These data lead us to believe that the scintigraphic techniques may be more sensitive in detecting early mandibular involvement with squamous cell carcinoma than the Panorex technique and may help alter the therapeutic approach. PMID- 6290963 TI - Dental analgesia produced by non-painful low-frequency stimulation is not influenced by stress or reversed by naloxone. AB - The dental pain threshold elevation produced by non-painful, low-frequency transcutaneous electrical nerve stimulation (TENS) in healthy humans was not reduced by the administration of 0.8 mg of naloxone i.v. Neither ACTH, prolactin nor growth hormone (GH) release were related to the pain threshold elevations. The present study indicates that the dental pain threshold elevation during non painful, low-frequency TENS is not based on the same opioid-dependent mechanisms as the dental pain threshold elevation during acupuncture or the clinical analgesia during low-frequency TENS. Stress or other adenohypophyseal mechanisms involving ACTH, prolactin or GH do not explain the analgesia induced by non painful, low-frequency TENS. PMID- 6290964 TI - Asbestos-related pleuropulmonary diseases: asbestosis, mesothelioma and lung cancer. PMID- 6290965 TI - Detection of tissue CEA-like substance as an aid in the differential diagnosis of malignant mesothelioma. AB - Sections of various adenocarcinomas and malignant mesotheliomas were tested for carcinoembryonic antigen (CEA) localized in tissues by the immunoperoxidase technique; epithelial mucin was demonstrated with the PAS technique. While CEA and mucin were found in many adenocarcinomas, both were absent in the 43 cases of malignant mesothelioma we investigated. In the problem of distinguishing between adenocarcinoma and mesothelioma, the CEA-test in combination with conventional strains for mucin is a useful technique and clearly identifies most adenocarcinomas. A dual negative result for CEA and mucin, although not proving that a given lesion is a mesothelioma, adds considerable support to this histological diagnosis. PMID- 6290967 TI - Mucin secreting cancer with mesangial IgA deposits. AB - Subclinical immune complex glomerular deposits occurred in 20% (11 of 55) of cancer cases examined postmortem. Eight of 11 (72.7%) mucin secreting adenocarcinomas of lung, stomach, large intestine and pancreas showed glomerular mesangial deposits of IgA, with associated IgM and C3 in one-half of them. The glomerular morphology showed predominantly minimal or minor changes, and less frequently mesangial cell proliferation. None of the patients exhibited clinical evidence of glomerulonephritis. The nature of the antigen(s) in the paraneoplastic phenomenon was not defined, but there may have been environmental antigen(s) or specific immune response in the local population to explain the IgA deposition. PMID- 6290966 TI - Symposium: classification of leukemia. 1. The classification of acute leukemia. AB - Two main forms of acute leukemia have been recognized by the French-American British (FAB) group: myeloid (AML) and lymphoblastic (ALL). Some types of AML can be diagnosed on well prepared bone marrow films stained with May-Grunwald-Giemsa. Poorly differentiated types, myeloblastic (M1) and monoblastic (M5PD), need confirmation by positive cytochemical reactions (Sudan Black B, myeloperoxidase and non-specific esterase). There are 2 sub-types of promyelocytic leukemia: M3 typical, hypergranular and M3 variant, microgranular. The M3 variant has a more acute course, higher WBC and may require cytochemistry to demonstrate promyelocytic differentiation. Electron microscopic cytochemistry can also help in the classification of difficult AML cases; the 'platelet-peroxidase' reaction, for example, is essential for the diagnosis of megakaryoblastic leukemia, a disorder often presenting as 'acute' myelosclerosis. Three morphological types are seen in ALL: L1, predominantly in children, L2, more frequently in adults, and the relatively rare L3 or Burkitt type. Immunological and enzyme markers (ALL and la antigens, terminal transferase, etc.) help define the cell phenotype: (1) non-B, non-T ALL with 3 forms (common, null and pre-B), (2) T-ALL, related to but distinct from T-lymphoblastic lymphoma, and (3) B-ALL, usually with L3 morphology, There is growing evidence that the FAB morphological types correlate with prognosis in ALL independently of other factors. The immunologically defined types also correlate with prognosis but not as an independent variable. PMID- 6290968 TI - [Pathogenesis of leukemias induced in XVII Bln mice by two types of oncorna virus]. PMID- 6290969 TI - [Ultrastructural and histocytochemical studies of the liver and kidneys in experimental lead acetate poisoning]. PMID- 6290970 TI - [Liver cell carcinoma--a comparison of the cytological and histological pictures]. PMID- 6290971 TI - Aspiration cytology of the breast. II. Significance of bipolar naked nuclei. PMID- 6290972 TI - The clinical value of CT whole-body scanning in pediatric tumour investigation. AB - The results of 100 consecutive CT whole-body scans in children with suspicion of a tumour and possible malignancy have been analysed. The final diagnoses were these: a tumour in 40 patients; non-tumoural lesions such as cysts, abscesses or malformations in 22; various medical conditions in 30 cases; and no abnormality in eight. The suspicion of malignancy was confirmed in 36 patients. CT proved to be an accurate method in the diagnosing and differentiation between solid or predominantly solid tumours and various non-tumoural lesions. The clinical value of CT was assessed by comparing the information from CT with those obtained by routine clinical and other radiological means. CT was positive informative in 57% of the patients and correctly excluding a tumour in 28%. CT was either non contributory or misleading in 15%. Finally, the quality of the scans were evaluated and the specific problems related to pediatric CT are discussed. PMID- 6290973 TI - Swyer-James-Macleod syndrome: a case with a baseline normal chest radiograph. PMID- 6290974 TI - Use of restriction enzymes to investigate the source of a primary cytomegalovirus infection in a pediatric nurse. AB - The risk of transmission of cytomegalovirus (CMV) infection from congenitally infected infants to nonimmune medical attendants is unknown. The case of a CMV seronegative, pregnant nurse who seroconverted after caring for an infant with symptomatic CMV infection is reported. She elected to be aborted and the fetal tissue contained CMV. Isolates from the nurse, the fetal tissue, and the infant to whom the nurse was exposed were examined for genetic relatedness by restriction enzyme analysis. As expected, the isolates from the nurse and the fetal tissue were identical. However, the virus isolated from the symptomatic infant was different from the strain infecting the nurse. These data indicate that the nurse acquired her infection from a source other than the index infant, either within the hospital or within the community. PMID- 6290975 TI - Restriction endonuclease analysis of cytomegalovirus deoxyribonucleic acid as an epidemiologic tool. AB - The possibility of transmission of cytomegalovirus (CMV) from congenitally infected infants to susceptible medical personnel produces anxiety because the risks have not been defined. A physician conceived her first child while caring for an infant in the intensive care nursery who died with congenital CMV infection. The physician had serologic evidence of active CMV infection confirmed by isolation of virus from multiple sites. She elected to have her pregnancy interrupted. CMV was isolated from the placenta and fetal brain. Restriction enzyme analysis was employed to test all the CMV isolated for genetic relatedness. Virus isolated from the physician and her fetus was identical. The virus from the index nursery infant was different from the strain infecting the physician and her fetus. These data indicate that the physician acquired her virus from a source other than the index infant. PMID- 6290976 TI - Epstein-Barr virus (EBV)-antibody profile in an infant congenitally infected by the EBV. PMID- 6290977 TI - [Cyclic nucleotide content in the peripheral blood leukocytes in acute leukemia in children]. PMID- 6290978 TI - [Latent dissemination of nephroblastomas]. PMID- 6290979 TI - [Characteristics of the cyclic nucleotide system in allergic states]. PMID- 6290980 TI - [Contribution of pirprofen to the medical treatment of radicular neuralgia]. AB - A multicentric therapeutic double blind study, conducted according to the incomplete block method and including 108 observations from 5 hospital centers, was conducted among patients suffering from acute radicular, sciatic, cervico brachial or crural neuralgia. The purpose was to compare the analgesic effect of 4 treatments given in single dose: pirprofen 200, pirprofen 400, ketoprofen 150 and placebo. Overall analysis showed a significant decrease in pain during the 4 hours following administration of the products and in all groups. However, pirprofen 400 and ketoprofen 150 mg caused a decrease in painful phenomena, more intense than that induced by pirprofen 200 and placebo. The great heterogeneity among the different treatment centers led the investigators to carry out a second analysis limited to the 4 most homogeneous centers. This analysis, of 84 observations, shows clearly the analgesic efficacy of both doses of pirprofen and ketoprofen compared to placebo. It also emphasizes the rapidity of action of pirprofen compared to the other products. The overall tolerance of the tested products was good. In the treatment of acute radicular neuralgia, pirprofen 400 mg is active in comparison to the placebo and more quickly effective than ketoprofen 150 mg. PMID- 6290981 TI - [Platelet prostaglandins and apparent components in diabetes mellitus]. PMID- 6290982 TI - Chemical synthesis and cloning of a gene for human beta-urogastrone. AB - A DNA duplex coding for the 53 amino acids of human beta-urogastrone has been synthesised. Computer assisted design of the gene included restriction endonuclease sites for plasmid insertion, a termination codon and two triplets coding for lysine at the 5'-end of the structural gene. The synthesis involved preparation of 23 oligodeoxyribonucleotides by phosphotriester procedures coupled to rapid HPLC techniques. The gene was constructed in two halves by enzymatic ligation of the oligonucleotides and cloned into a specially constructed chimeric plasmid vector. Escherichia coli K12 MRC8 was transformed by the plasmid and clones containing the full gene sequence were isolated and characterised. PMID- 6290983 TI - Sequence variation and methylation of the flax 5S RNA genes. AB - The complete sequence of the flax 5S DNA repeat is presented. Length heterogeneity is the consequence of the presence or absence of a single direct repeat and the majority of single base changes are transition mutations. No sequence variation has been found in the coding sequence. The extent of methylation of cytosines has been measured at one location in the gene and one in the spacer. The relationship between the observed sequence heterogeneity and the level of methylation is discussed in the context of the operation of a correction mechanism. PMID- 6290984 TI - A cloned immunoglobulin cDNA fragment enhances transposition of IS elements into recombinant plasmids. AB - Evidence is presented indicating that a novel DNA sequence arrangement generated by in vitro recombination may elicit high frequency transpositions of IS elements. A 109 bp Bam HI fragment of the cDNA for the immunoglobulin kappa light chain from MOPC 321 myeloma was cloned into the Bam HI site of pBR313. The cloned fragment extends from the codon for Gly 57 to the V-J junction. Insertions of IS1 or IS5 were identified in 6 of 50 plasmid DNAs isolated from freshly transformed clones. Additional transposition events were detected after subculturing for several growth cycles. Three independent insertions of IS1 occurred in the promoter region of the TcR operon. All IS5 and the remaining IS1 insertions were located in the TcR region upstream to the cloned DNA sequence. Sequences homologous to the ends of IS1, or corresponding to the consensus sequence at the target site of IS5 are present near the estimated sites of insertion of IS1 or IS5 respectively. Bacteria harboring recombinant plasmids carrying the cloned DNA in either orientation grew at a reduced rate relative to cells harboring pBR313, suggesting that fused gene products made from the two types of plasmid were inhibitory to cell growth. IS insertions, which relieved this inhibitory effect and thereby provided a selective advantage, were found exclusively in plasmids carrying the cloned DNA in only one of the two orientations. The fact that IS elements were not observed in the other type of recombinant plasmid indicates that selective pressure alone is not sufficient to account for the frequent IS insertions observed and that sequences at a distance from the site of IS insertion may be critical in the regulation of transposition frequency. PMID- 6290985 TI - Spliced and unspliced virus specific RNA sequences are associated with purified simian virus 40 chromatin. AB - We have analyzed SV40 specific RNA sequences which are associated with purified viral chromatin. Three discrete size classes of large unprocessed transcripts were identified: 4.8, 3.6 and 3.0 kb. Furthermore, two polyadenylated species of 2.6 and 1.6 kb were found, the latter of which representing mature 16 S mRNA. S1 analysis revealed both the presence of spliced and unspliced RNA sequences in transcriptionally active SV40 chromatin. PMID- 6290986 TI - Nucleotide sequence of a protamine component CII gene of Salmo gairdnerii. AB - We have isolated, using nick-translated cloned protamine cDNA's as probes, several genomic clones containing protamine gene sequences from a Charon 4A library of Eco R1 digested rainbow trout (Salmo gairdnerii) DNA. One clone was chosen for detailed study and the 2.5 kbp Bam HI-Eco R1 restriction fragment containing the gene was subcloned in the plasmid pBR322. A 920 bp Bg1 II - Bam HI restriction fragment contains a sequence coding for protamine component CII as well as regions 5' and 3' to the mRNA coding portion. Present in the region 5' to the mRNA coding sequence are the promoter associated signals "TATA" box and "CAAT" box. The 5' untranslated region of the mRNA whose length and sequence were not established from the cDNA clones (1) was determined by nuclease mapping and starts within a sequence similar to the "capping signal" found in other genes. The protamine gene for CII contains no introns, a situation common to most histone genes, but, unlike the histone genes does not occur close to other protamine genes in a "cluster". PMID- 6290987 TI - Expression of a X. laevis tRNATyr gene in mammalian cells. AB - Expression of a X. laevis tRNATyr gene has been studied in mammalian cells. This tRNATyr gene has a 13 base intervening sequence adjacent to its anticodon. A fragment containing the tRNATyr gene was cloned into the late region of SV40. Cells infected with a recombinant virus stock vastly overproduce a tRNATyr that is properly spliced, processed and modified. It was also found that the X. laevis tRNATyr is identical or nearly identical to an endogenous tRNATyr of monkey kidney cells. The possibility of using the X. laevis tRNATyr gene to create an amber suppressor for mammalian cells is discussed. PMID- 6290988 TI - A high speed, high capacity homology matrix: zooming through SV40 and polyoma. AB - We present a new homology matrix program which owes its basic conception to the two-dimensional dot matrices previously described (1,2), but has important improvements and new features. It scores sequence homology over an adjustable range and plots the scores which are above an operator-determined filtration level. Its powerful noise-filtration system, capacity for compression without much loss of information, and speed of execution make this program a valuable tool in the analysis of homologies, internal direct repeats and reverse repeats, including palindromic sequences. The properties of the program are exemplified by analysis of SV40 and polyoma DNA sequences. PMID- 6290989 TI - Nucleotide sequence of Aspergillus nidulans mitochondrial genes coding for ATPase subunit 6, cytochrome oxidase subunit 3, seven unidentified proteins, four tRNAs and L-rRNA. AB - The complete nucleotide sequence of a 14 kb segment of A. nidulans mtDNA reveals a rather compact organization of genes transcribed from the same strand and coding for two functionally known proteins, seven unidentified polypeptides (URFs), 24 tRNAs and two rRNAs. One of the URFs is located in the intron of the L rRNA gene and codes for a basic protein of 410 residues. The other URFs are in spacer regions and code for hydrophobic proteins. URFa is homologous to human URF4, and URFb produces a polypeptide of 48 residues resembling the human URF6L product (hydrophobic N-terminus, basic C-terminus). The ATPase subunit 6 genes from mitochondria and E. coli appear to share a common ancestor. The codon frequencies of identified genes and URFs are similar, and codons ending with G or C are rarely used. The structures of tRNAs specific for arginine, asparagine, tyrosine and histidine are deduced from gene sequences. PMID- 6290990 TI - Use of a cellular polyadenylation signal by viral transcripts in polyoma virus transformed cells. AB - The DNA sequences at and around the junctions between viral and cellular DNA in the polyoma virus transformed mouse cell line, TS-A-3T3, have been determined. No common sequence specificity or structural features at the joins have been observed. The sequence indicates that the 94K truncated large T antigen found in TS-A-3T3 cells is a hybrid protein in which the carboxy-terminal 19 amino acids are encoded by adjacent host sequences. Moreover, the three early region transcripts initiated in viral sequences are also hybrid in nature and appear to utilize a host polyadenylation signal associated with the hexanucleotide, AATAAA, found 100 bp beyond a viral-host join. PMID- 6290991 TI - Isolation of the human HDL apoprotein A1 gene. AB - Apo A1 is the major apoprotein of the human plasma high density lipoprotein (HDL). We have isolated apo A1 cDNA and genomic clones and used them to study the gene organisation as defined by its restriction enzyme map. These studies showed that apo A1 is coded by a unique gene. Cross hybridisation was not observed with functionally related apoprotein genes. Increased levels of HDL have been correlated with certain protection against coronary heart disease. If there is a genetic component that contributes to the variable levels of HDL found in the population, it may be possible to correlate these differences with distinct gene organisation patterns. PMID- 6290992 TI - The expression of biologically active cholera toxin in Escherichia coli. AB - Chromosomal DNA from Vibrio cholerae El Tor strain 1621 was digested with Hind III and the products fractionated by centrifugation through a sucrose gradient. A 15kb fragment containing the toxin gene of V. cholerae was identified by its homology with the heat labile toxin (LT) gene of toxigenic E. coli. This fragment was cloned in E. coli using pAT153 and subsequently characterised by digestion with different restriction endonucleases. Sequences homologous to the LT gene were identified by hybridisation and then sub-cloned using either pAT153 or pACYC184. Expression of the cloned CT gene in E. coli was detected using both cell culture and ELISA assays. One recombinant plasmid coded for the synthesis of an immunologically active but biologically inactive derivative of CT. PMID- 6290994 TI - Identification of novel DNA forms in tomato golden mosaic virus infected tissue. Evidence for a two component viral genome. AB - Extracts obtained from cells infected with the geminivirus tomato golden mosaic (TGMV) are shown to contain, in addition to viral single-stranded DNA, several novel species of virus-specific single- and double- stranded DNA (ss and ds DNA). The results of nuclease studies and electron microscopy suggest that three of the intracellular DNAs are unit-genome length duplexes of closed circular, relaxed circular, and linear form. The remaining ds DNA species are of high molecular weight and appear to be concatamers consisting of two or more unit-length circular ds TGMV DNA resulted in fragments whose combined size is twice the unit genome length. Thus ds TGMV is composed of two components of nearly identical size but different nucleotide sequence. PMID- 6290993 TI - Isolation and nucleotide sequence of a mouse histidine tRNA gene. AB - We have sequenced a 1307 base pair mouse genomic DNA fragment which contains a histidine tRNA gene. The sequence of the putative mouse histidine tRNA differs from the published sequence of sheep liver histidine tRNA by a single base change in the D-loop. It does not contain an unpaired 5' terminal G residue, as reported for Drosophila and sheep histidine tRNAs. The gene does not contain introns. The 3' flanking region contains a typical RNA polymerase III termination site of 6 consecutive T residues. 523 residues after the 3' end of the his tRNA coding region, the mouse DNA contains a sequence 72% homologous to part of the consensus sequence of the B1 (alu) family. PMID- 6290995 TI - Molecular cloning and characterisation of the two DNA components of tomato golden mosaic virus. AB - We report the molecular cloning of the tomato golden mosaic virus (TGMV) genome in the E. coli plasmid pAT 153. The results of this work conclusively show that TGMV DNA consists of two components (designated A and B) of almost, but not exactly, the same size. Four different recombinant plasmids are described, two containing component A in opposite orientation and two containing component B in opposite orientation. Southern blot analysis revealed little sequence homology between A and B and showed both components to be equally represented in viral and intracellular DNA forms. Detailed restriction maps of the cloned DNAs are presented, and a comparison of these with digests of intracellular viral dsDNA indicates that the former are full-length faithful copies of TGMV DNA. This is the first report of the cloning of a geminivirus genome. PMID- 6290996 TI - Sequence of the intercistronic region between the ribulose-1, 5-bisphosphate carboxylase/oxygenase large subunit and coupling factor beta subunit gene. AB - The nucleotide sequence of the region between the genes for the large subunit of ribulose-1, 5-bisphosphate carboxylase/oxygenase and the beta subunit of coupling factor from tobacco chloroplast DNA has been determined. The 5' ends of their mRNAs have been located on the DNA sequence using s1 nuclease mapping. The promoter sequences of these chloroplast genes have features in common with those of prokaryotes. E. coli RNA polymerase can recognize these sequences and start transcription at authentic initiation sites. PMID- 6290997 TI - Sequence requirements for the transcription of the rabbit beta-globin gene in vivo: the -80 region. AB - We have performed a detailed analysis of DNA sequences in the -80 region of the rabbit beta-globin gene that are required for transcription. A variety of rabbit beta-globin gene templates deleted at various sites in the -80 region were linked to an SV40-plasmid recombinant and introduced into HeLa cells; under these conditions the rabbit beta-globin gene is expressed from its own promotor. The results show that the conserved GGCCAATCT sequence is required for efficient transcription in vivo and further identify another sequence in the region from about -81 to -96 which is also required for transcription in vivo. PMID- 6290998 TI - The maize chloroplast genes for the beta and epsilon subunits of the photosynthetic coupling factor CF1 are fused. AB - We have cloned and sequenced the maize chloroplast genome fragment Eco RI e which contains the 2.2 kb transcript previously reported (Link, G. and Bogorad, L. (1980) Proc. Nat. Acad. Sci. 77 6821-6825) to lie next to the maize gene for the large subunit of ribulose bisphosphate carboxylase (LS) and to be transcribed divergently. Immunochemical and sequencing data show that the gene codes for the beta subunit of the maize chloroplast coupling factor complex (CF1). The derived amino acid sequence is highly homologous to that of the corresponding E. coli protein (Saraste et al. (1981) Nucleic Acids Res. 9 5287-5296). The last base of the codon for the terminal lysine residue of the beta subunit of CF1 is the first base of the codon for the initiating methionine of an open reading frame whose derived amino acid composition and size closely match that reported for the epsilon subunit (Binder et al. (1978) J. Biol. Chem. 253 3094-3100). The close coupling of the two genes may serve to in sure their stoichiometric production. PMID- 6290999 TI - Characterization of a highly repetitive family of DNA sequences in the mouse. AB - A large proportion (0.5-1%) of total mouse DNA is cleaved by Bam HI into fragments whose size is about 500 base pairs. A cloned member of this repetitive family of DNA sequences (BAM5 family) was sequenced by the dideoxy chain termination procedure and shown to contain 507 base pairs. The sequence exhibited no unusual or remarkable features. Repetitive sequences complementary to the cloned BAM5 fragment were found in rat DNA, but not in feline or human DNA. Restriction mapping suggested that many BAM5 sequences were components of much larger repetitive DNAs which were scattered throughout the mouse genome. The BAM5 sequences within the larger repetitive DNAs did not appear to be arranged tandemly or as members of scrambled tandem repeats. RNA homologous to the cloned BAM5 sequence was detected in cultured mouse cells, but not in cultured rat cells. PMID- 6291000 TI - Transcription initiation sites within an IS2 insertion in a Gal-constitutive mutant of Escherichia coli. AB - Insertion of the insertion sequence Is2(I) directly before the galE gene of the galactose operon results in a Gal minus phenotype (1, 2). The Gal-constitutive allele galc200 (and its deletion derivative galc200 delta 31) arise from such a Gal minus mutant by the insertion of LS2(II) DNA within the LS2(I) sequence (3). We have transcribed in vitro a DNA template representing the IS2-galE region of galc200 delta 31. Gal-directed transcription initiates at two sites within the IS2(I) sequence, 51 and 52 bp from the IS2-galE junction. The promoter for these transcripts, Pgal200 delta 31, is composed of a novel joint between a -10 region from the IS2(I) DNA and a -35 region contributed by the IS2(II) insertion. No promoters intrinsic to the 121 bp of the IS2(II) sequence also present on the template were detected. The relevance of Pgal200 delta 31 to the Galc phenotype of galc200 and to general mechanisms for the constitutive expression of genes adjacent to IS2 is discussed. PMID- 6291001 TI - PpGpp regulates the binding of two RNA polymerase molecules to the tyrT promoter. AB - Bacterial promoters differ in the number of RNA polymerase molecules that bind to form a filterable polymerase-promoter complex. We show that two holoenzyme molecules interact with the tyrT promoter, probably as a dimer. This interaction is inhibited by ppGpp. By contrast a single holoenzyme monomer suffices for complex formation at the lacUV5 promoter. We propose that In vivo promoter selection by monomeric and dimeric forms of the enzyme could coordinate the synthesis of stable RNA with that of mRNA and could also account in part for the switch in transcriptional selectivity during the stringent response. PMID- 6291004 TI - The TCDD receptor in rat intestinal mucosa and its possible dietary ligands. AB - Induction of aryl hydrocarbon hydroxylase (AHH) by polycyclic aromatic hydrocarbons and other inducers such as 2,3,8-tetrachlorodibenzo-p-dioxin (TCDD) is known to occur following binding of the inducer to a soluble receptor protein similar to steroid hormone receptors. This receptor is usually called the TCDD receptor, since TCDD has the highest affinity of all known ligands for the receptor. In the present paper a receptor for TCDD in cytosol from rat intestinal mucosa has been studied, using isoelectric focusing in polyacrylamide gel. This receptor's biochemical properties were found to be similar to those of the TCDD receptor in rat liver cytosol. The dissociation constant (Kd) of the 3H-TCDD receptor complex in rat intestinal mucosa was 0.7-3.1 nM, and it was present at a concentration of 70-80 fmol/mg protein. Starvation did not significantly increase the receptor level. The affinities of some potential dietary ligands for the TCDD receptor in rat intestinal mucosa were also studied. Indole-3-carbinol had 1/2,600 of the affinity of TCDD for the receptor protein. Butylated hydroxyanisole (BHA), transstilbene oxide and quercetinpentamethylether competed even more weakly with 3H-TCDD for binding to the receptor. The biological significance of the occurrence of low-affinity ligands of dietary origin for the TCDD receptor is uncertain at the present time. PMID- 6291005 TI - Absence of sperm-specific lactate dehydrogenase-x in patients with testis cancer. AB - LDH-x is a unique form of lactate dehydrogenase found in germinal epithelium during spermatogenesis. 53 patients with germ cell tumors were screened for the presence of LDH-x in the serum. No patient had detectable levels of the enzyme. 31 of the patients had elevated total serum LDH levels, and the isozyme fraction type 1 and/or type 2 was responsible for the elevation in all cases. LDH is a useful marker in germ cell tumors, but LDH-x is not the responsible isozyme. PMID- 6291003 TI - Increased activity of polynucleotide ligase in 5-iodo-2'-deoxyuridine and mitomycin C-pretreated simian virus 40 (SV40)-infected monkey kidney cells. AB - The DNA ligase activity has been studied in 5-iodo-2'-deoxyuridine (IUdR), mitomycin C (MMC) and IUdR + MMC-treated SV40-infected or mock-infected CV1C11 monkey kidney cells. The results have shown that : 1. The level of enzyme activity is about two or three times higher in IUdR or MMC-treated non-infected cells; 2. SV40 infection doubles the level of ligase activity in the untreated cells; 3. There is an additional increase of ligase activity in IUdR or MMC treated SV40-infected cells; 4. When infected or mock-infected cells are treated with IUdR + MMC, there is no modification of the results obtained with each drug alone; 5. Two peaks of different S values are detected in a partial purification of the drug(s) or viral induced enzyme(s). The increase in the ligase activity in drug-treated cells is independent of semi-conservative DNA synthesis. The drug(s) and viral-induced enzyme(s) is the consequence of a "de novo" synthesis of proteins, as shown by cycloheximide treatment. PMID- 6291002 TI - Influence of nonhistone chromatin protein HMG-1 on the enzymatic digestion of purified DNA. AB - The effect of chicken erythrocyte High Mobility Group protein 1 (HMG-1) on the enzymatic hydrolysis of purified double-stranded and single-stranded bacteriophage lambda DNA was studied. HMG-1 was found to inhibit the digestion of single- and double-stranded DNA by S1 nuclease and DNase I, respectively. HMG-I increased the rate of hydrolysis of double-stranded DNA by micrococcal nuclease, particularly at low HMG-1/DNA ratios, and had little effect on the hydrolysis of single-stranded DNA by micrococcal nucleases, even at high HMG-1 DNA ratios. We also present a semi-quantitative estimate that HMG-1 and HMG-2 occur in chromatin from rapidly dividing, cultured rat hepatoma cells at about 8 times the level that they occur in adult rat liver chromatin. PMID- 6291006 TI - Tumor marker production in human bronchial carcinoma xenografts. AB - A total of 37 human bronchial carcinomas (15 small cell, 10 squamous, six large cell and six adenocarcinomas) have been successfully heterotransplanted into immune-suppressed mice. Monolayer tissue cultures have been established from 23 of the xenograft lines. After contact with tumour cells, supernatant culture medium was analysed by radioimmunoassay for adrenocorticotrophin (ACTH), calcitonin, human chorionic gonadotropin beta subunit (beta HCG) and carcinoembryonic antigen (CEA). Calcitonin production was associated with three (13%) xenograft lines (two small cell, one adenocarcinoma). beta HCG was found in three (13%) lines (one small cell, two large cell anaplastic). CEA was produced in 13 (56.5%) lines (seven small cell, two squamous, one large cell, three adenocarcinomas). ACTH was present in 18 (78.2%) lines (11 small cell, two squamous, two large cell, three adenocarcinomas), but evidence indicates that this hormone is not in all cases produced and secreted by the tumour. PMID- 6291007 TI - Monitoring therapy in trophoblastic diseases by radioimmunoassay of pregnancy specific beta 1-glycoprotein and the beta subunit of human chorionic gonadotropin. AB - The beta-subunit of human chorionic gonadotrophin (hCG) and pregnancy-specific beta 1-glycoprotein (SP1) were measured simultaneously by specific radioimmunoassays in 473 serum samples from 43 patients with trophoblastic tumors. Both SP1 and hCG values were high in 16 untreated patients. 27 patients had been treated previously elsewhere, and their further treatment and monitoring were started simultaneously in our institute. Raised SP2 and hCG levels were found in 22 and 21 cases, respectively. Treatment was followed by a parallel decrease of both SP1 and hCG, but the hCG decline was more marked. Seronegativity was reached only after several months; SP1 was present in the serum for 60-150 days, and hCG could be found even after this time. Both parameters were sensitive indicators of residual tumor activity, calling for further therapy months before any clinical manifestation of recurrence. In three cases only one of the tumor makers could be found, which underlines the importance of the simultaneous measurement of hCG and SP1. PMID- 6291008 TI - Effects of CCK on gastrointestinal function in lean and obese Zucker rats. AB - Cholecystokinin (CCK), a hormone affecting several gastrointestinal functions, has also been shown to elicit satiety and affect daily meal patterns. Since Zucker obese rats are less sensitive to the satiety effects of CCK, two experiments were designed to determine if they are also less sensitive to the gastric emptying and intestinal transit rate effects of CCK. In the first experiment phenol red was administered to 5.5 hr fasted rats 15 minutes after intraperitoneal injection of CCK-8 or saline. Rats were sacrificed after 30 minutes, the stomach and small intestine were removed, and phenol red content was measured. More phenol red was in the stomach of obese but not lean rats treated with CCK-8. The rate of transit of the contents of the small intestine was increased by CCK-8 and the percent of phenol red in the fourth quarter of the small intestine was greater in obese than lean rats (91 vs 37%, p less than 0.05). In the second experiment gastrointestinal transit of ferric oxide was measured during the light and dark phases of the diurnal cycle, and when obese rats were ad lib or yoke-fed to lean pair-mates. Total gastrointestinal transit time of the ferric oxide was decreased 15% when CCK-8 was administered to yoke fed obese rats in either the light or dark portions of the diurnal cycle but was not affected in ad lib-fed obese rats or lean rats. Thus, while Zucker obese rats are less sensitive to satiety effects of CCK, they appear to be more sensitive to the gastrointestinal effects of CCK, and therefore it is not clear what role these gastrointestinal responses have on the feeding behavior responses. PMID- 6291009 TI - An analog of ACTH/MSH (4-9), ORG-2766, reduces cerebral uptake of morphine. AB - The uptake of morphine was significantly reduced in most regions of the brains of conscious, unrestrained rats within 10 minutes after treatment with an analog of ACTH/MSH (4-9), ORG-2766. The effect was most obvious in regions with significant densities of enkephalin receptors, namely basal ganglia, hippocampus and cortex. The results explain, in part, how some fragments and analogs of ACTH/MSH may antagonize behavioral actions of morphine, even though some of these peptides lack significant opiate receptor binding properties. We believe that this effect of ORG-2766 is related to an action on the permeability characteristics of the brain microvasculature. The underlying mechanism is unknown. PMID- 6291011 TI - Morphine, beta-endorphin and [D-Ala2] Met-enkephalin inhibit oxytocin release by acetylcholine and suckling. AB - Morphine inhibits suckling-induced oxytocin (OT) release in lactating mice. Since beta-endorphin and enkephalins have several actions in common with morphine, the action of these opioid peptides on OT release was investigated. In anesthetized lactating rats, OT release was achieved by intraventricular injection of acetylcholine (ACh) or by the physiological stimulus of suckling. The amount of OT released was estimated by comparing milk-ejection responses to these stimuli to those following known amounts of intravenous (IV) OT. Both beta-endorphin and [D-Ala2]Met-enkephalin inhibited ACh-induced and suckling-induced OT release. Naloxone antagonized opiate inhibition in both cases. PMID- 6291010 TI - Neurons containing alpha-melanocyte stimulating hormone and beta-endorphin immunoreactivity in the cat hypothalamus. AB - Immunohistochemical studies were conducted on sections of cat hypothalamus in order to determine the distribution of neurons containing alpha-melanocyte stimulating hormone and beta-endorphin immunoreactivity. A large number of neurons in the arcuate nucleus were stained after incubation of sections with antisera to either substance. Analysis of serial sections suggested that each neuron revealed with one antiserum was also revealed with the other antiserum, indicating the co-existence of alpha-melanocyte stimulating hormone and beta endorphin immunoreactivity within these arcuate neurons. In contrast, a more diffuse group of lateral hypothalamic neurons which extended from the retrochiasmatic level to the posterior hypothalamus were stained only with the antiserum directed against alpha-melanocyte stimulating hormone. The present results largely confirm findings in the rat hypothalamus, although the lateral hypothalamic group of alpha-melanocyte stimulating hormone immunoreactive neurons appears to be more extensive in the cat. PMID- 6291013 TI - Pineal mediation of the thermoregulatory and behavioral activating effects of beta-endorphin. AB - Intraventricular administration of the opioid peptide, beta-endorphin to goldfish altered their body temperatures and activity levels. Low doses (0.5-5.0 pg g-1 body weight) of beta-endorphin significantly increased behaviorally selected body temperatures while higher doses (15 pg g-1) decreased the preferred temperatures selected in horizontal thermal gradients. There was a significant day-night rhythm in the extent of these effects. These thermoregulatory effects could be blocked and reversed by systemic administration of the opiate antagonist, naloxone, supporting mediation of the thermoregulatory effects at opioid receptors. In addition, administration of naloxone by itself significantly decreased preferred temperature. Removal of the pineal gland significantly increased the preferred temperatures selected by goldfish and eliminated the thermoregulatory effects of beta-endorphin administration in both the day and the night. The behavioral activity effects of beta-endorphin were dependent on the thermal conditions. In fish held at a constant temperature (20 degrees C) beta endorphin caused a dose-dependent increase in activity, while in individuals held in thermal gradients administration of beta-endorphin had no effects on activity. In both situations naloxone caused a decrease in activity levels. Pinealectomy also eliminated the behavioral activating effects of beta-endorphin, though it had no apparent effects on the actions of naloxone. These results indicate that the pineal gland is involved in the mediation of the thermoregulatory and behavioral activating effects of beta-endorphin. Speculations are made as to the possible mechanisms of action of the pineal gland in mediating the effects of opioid neuropeptides. PMID- 6291014 TI - Ceruletide, ceruletide analogues and cholecystokinin octapeptide (CCK-8): effects on motor behaviour, hexobarbital-induced sleep and harman-induced convulsions. AB - Ten ceruletide analogues and cholecystokinin octapeptide (CCK-8) were compared with ceruletide regarding neuropharmacological effects in mice after subcutaneous administration. The effects under study were catalepsy, prolongation of hexobarbital-induced sleep and delay in onset of harman-induced convulsions. Ceruletide and several analogues were more potent than the reference drugs, diazepam and haloperidol. Desulfation, deamidation and shortening of the peptide chain by five amino acids strongly reduced or abolished the pharmacological activities of ceruletide. Other chemical modulations mostly weakened the efficacy, but to an unequal extent for the three effects, which altered the pharmacological selectivity. PMID- 6291012 TI - Characteristics of thyrotropin releasing hormone (TRH) receptors in rat brain. AB - Characteristics of TRH-receptors were studied in the rat central nervous system (CNS). Ion species, pH and temperature importantly influenced TRH-receptor binding. Subcellular distribution of TRH-receptor binding revealed that synaptic membranes had the greatest percentage of total sites. Scatchard analysis suggested that the rat CNS had two distinct TRH binding sites with apparent dissociation constants (Kd) of 5 X 10(09) M and 13 X 10(-8) M. Receptor activity is sensitive to trypsin and phospholipase A digestion, suggesting that protein and phospholipid moieties are essential for the binding of [3H]TRH. Thiol reagents reduced the binding activity of the receptor, suggesting that an intrachain disulfide bond may form an important constituent of the binding site for TRH. The TRH-receptor in the rat brain was successfully solubilized with non ionic detergent Triton X-100. On gel chromatography with Sepharose 6B column, the solubilized TRH-receptor molecule eluted at the fraction corresponding to an apparent molecular weight of 300,000 daltons, with Stokes' radius of 5.8 nm. The regional distribution of TRH-receptor binding was examined to clarify the site of TRH action. The highest level of binding was in the hypothalamus, cerebral cortex and hippocampus, indicating that TRH affects the CNS function mainly through the limbic system, cerebral cortex and hypothalamus. Moreover, tricyclic anti depressants and Li+ decreased the binding of [3H]TRH. These findings suggest that endogenous TRH and TRH receptor may play the role of a neurotransmission modulator in the brain to control emotional and mental functions. PMID- 6291015 TI - [Serum 5'-nucleotidase activity in chronic pancreatitis]. PMID- 6291017 TI - [Herpesviruses and chromosomes]. PMID- 6291016 TI - [Current theories on the mechanism of action of narcotic analgesics and endorphins]. PMID- 6291018 TI - [Morphine-like peptides and the digestive tract]. PMID- 6291019 TI - Isolation of high and low molecular weight components from chicken sera that have Rous sarcoma virus neutralizing activity. AB - Blood sera components from Arkansas regressor line (R-line) and progressor line (Pr-line) chickens are compared for the first time for Rous sarcoma virus neutralizing activity. Sera was fractionated by Sephadex G-100 filtration into a high molecular weight fraction I (HMW-I) and a low molecular weight fraction II (LMW-II) component (HMW-I greater than 14,000 daltons, LMW-II less than 5,000 daltons). Both fractions from each line of chickens exhibit activity against Rous sarcoma virus (RSV) judged by a wing web assay. Both HMW-I (principally antibodies) and LMW-II neutralized RSV when obtained from hyperimmune R-chickens and Pr-chickens with large progressing tumors. However, HMW-I and LMW-II obtained from R- or Pr-chickens before challenge contain so RSV neutralizing activity. The novel low molecular weight fraction II disappeared from the sera of R-line chickens 2 weeks after tumor regression, whereas the HMW-I persisted after tumor regression. PMID- 6291020 TI - Field trials with an oil emulsion infectious bursal disease vaccine in broiler breeder pullets. AB - Two independent field trials were undertaken with commercial broiler companies in separate sections of Alabama in order to test the efficacy of an experimental commercially prepared oil emulsion infectious bursal disease (IBD) vaccine in broiler breeder pullets. The IBD vaccine was tested both for safety and the production of neutralizing antibodies in pullets and their progeny as well as its ability to prevent early field exposure to virulent IBD virus (IBDV) in broiler progeny that may lead to immunosuppression resulting in poor performance. The following was demonstrated in both trials: 1) the IBD vaccine had no adverse effect on breeder performance, 2) the IBD antibody titers in breeders peaked at 10 weeks postvaccination (PV) and remained about 1.5 to 4 times higher than nonvaccinated breeders through 25 weeks (PV), 3) 1-day IBDV maternal titers in broilers were related to the parental titers, 4) the IBD vaccination in pullets prevented early infection to field IBDV in progeny as determined by antibody titers and various performance parameters. PMID- 6291021 TI - The effects of a pectic enzyme on the growth-depressing and rachitogenic properties of rye for chicks. AB - Rye has growth-depressing and rachitogenic properties when fed to chicks. Growth depression also is observed when chicks are fed certain polysaccharides, particularly pectin. Rye has about 8% pectin-like material. A commercially available pectic enzyme preparation was fed to chicks at .1% of the rye diet. This enzyme permitted better utilization of rye for growth and also alleviated the rachitogenic effects of rye. The supplemental enzyme permitted maximal bone mineralization with 200 to 300 ICU of vitamin D3/kg diet, whereas 500 ICU/kg otherwise were required with rye diets. PMID- 6291022 TI - [Neurologic diseases caused by cytomegalovirus]. PMID- 6291023 TI - The beta-adrenergic receptor in mammalian placenta: species differences and ontogeny. PMID- 6291024 TI - Identification of cloned Y chromosomal DNA sequences from a lampbrush loop of Drosophila hydei. AB - By recombinant DNA techniques, a Y chromosomal sequence of Drosophila hydei was isolated. This DNA sequence of 8.93-kilobase length is a member of a family of repetitive sequences located in the short arm of the Y chromosome. Tissue specific transcripts complementary to the cloned sequence were found in testes RNA. In situ hybridization demonstrated that such transcripts are present in the lampbrush loop pair "nooses" in primary spermatocyte nuclei--a loop pair that is associated with the only fertility gene in the short arm of the Y chromosome. PMID- 6291025 TI - cAMP stimulates transcription of the gene for cytosolic phosphoenolpyruvate carboxykinase in rat liver nuclei. AB - The effects of starvation, glucose refeeding, dibutyryl cAMP, and dexamethasone on expression of the gene for phosphoenolpyruvate carboxykinase (GTP) [GTP:oxaloacetate carboxy-lyase (transphosphorylating), EC 4.1.1.32] from rat liver cytosol was studied by using a cloned cDNA probe. The rate of transcription of the gene for phosphoenolpyruvate carboxykinase in hepatic nuclei isolated from starved rats decreased rapidly after refeeding with glucose. Administration of dibutyryl cAMP to glucose-refed animals increased the rate of phosphoenolpyruvate carboxykinase gene transcription seven-fold within 20 min. Phosphoenolpyruvate carboxykinase mRNA in the cytosol is 2.8 kilobases long whereas liver nuclei contain four precursor RNA species that are up to 6.5 kilobases long. Feeding glucose to starved rats rapidly decreased the sequence abundance of enzyme mRNA in both nuclei and cytosol. However, the decrease in cytosolic phosphoenolpyruvate carboxykinase mRNA was preceded by a transient increase in enzyme mRNA over the first 20 min after glucose refeeding. Administration of dibutyryl cAMP to glucose-refed starved animals increased the concentration of the nuclear RNA precursors of phosphoenolpyruvate carboxykinase five- to eight fold within 30 min and induced the mRNA for the cytosolic enzyme over a period of 60 min. We conclude that cAMP induces phosphoenolpyruvate carboxykinase mRNA by increasing the rate of gene transcription. PMID- 6291027 TI - Purified glucocorticoid receptor-hormone complex from rat liver cytosol binds specifically to cloned mouse mammary tumor virus long terminal repeats in vitro. AB - Purified glucocorticoid hormone-receptor complex (HRC) from rat liver binds to specific DNA sequences contained in cloned mouse mammary tumor virus (MMTV)DNA. The binding site of the hormone-receptor complex is located in the long terminal repeat (LTR) of MMTV DNA as shown by filter binding studies with labeled restriction fragments and by visualization of DNA-receptor complexes with the electron microscope. The DNAs from cloned MMTV lacking the LTR sequences were neither retained on nitrocellulose filters nor bound specifically to HRCs examined in the electron microscope. The HRC also failed to bind to restriction fragments from pBR322 and phage lambda. Specific binding of the HRC to LTR sequences is dependent upon occupancy of the receptor by a glucocorticoid. Previous work has demonstrated that the MMTV transcription is initiated within the LTR; additionally, MMTV transcription is known to be regulated by glucocorticoids. Our present results therefore support the hypothesis that HRC regulates hormone-induced transcription by binding to specific DNA sequences near the MMTV transcription start site. PMID- 6291026 TI - Methylation of simian virus 40 Hpa II site affects late, but not early, viral gene expression. AB - DNA methylation has been correlated with reduced gene expression in a number of studies, although evidence for a casual link between the two events has been lacking. Because microinjection of simian virus 40 (SV40) DNA into the nucleus of Xenopus laevis oocytes results in the synthesis of both early and late viral gene products, it was possible to test whether a specific methylation event can affect gene expression. The single SV40 Hpa II site at 0.72 SV40 map units was specifically methylated with Hpa II methylase. When this DNA was injected into oocytes, there was a marked reduction in the synthesis of the major late viral capsid protein VP-1, relative to the synthesis by an unmethylated control. However, production of the early proteins (the large and small tumor antigens) was not affected by Hpa II methylation. Therefore, methylation at a single site on the viral DNA located near the 5' end of the late region can specifically repress late gene expression. The possible mechanisms by which this repression is mediated are discussed. PMID- 6291028 TI - Hormone receptor modulates the regulatory component of adenylyl cyclase by reducing its requirement for Mg2+ and enhancing its extent of activation by guanine nucleotides. AB - N-Ethylmaleimide treatment of rat liver plasma membranes results in an adenylyl cyclase (EC 4.6.1.1) system that shows no measurable cyclizing activity but retains both an active glucagon receptor and a receptor-sensitive regulatory component N as assessed by reconstitution into cyclase-negative (cyc-) membranes from S49 murine lymphoma. Treatment of such N-ethylmaleimide-treated membranes, termed C- liver membranes, with guanosine 5'-[gamma-thio]triphosphate (GTP[gamma S] ) and Mg2+, followed by the removal of GTP[gamma S] by washing, yields an activated N which upon mixing with cyc- S49 membranes reconstitutes the cyc- S49 membrane adenylyl cyclase in the absence of added GTP[gamma S]. It was found that GTP[gamma S] activation of the N at saturating concentrations of GTP[gamma S] is slow at low Mg2+ concentration and accelerated by increasing Mg2+ concentrations. Addition of glucagon during the activation results in a lowering of the Mg2+ requirement for full activation from 25 mM to around 10 muM and in concomitant increases in both the rate and the extent of N activation. In contrast to its dramatic effect on Mg2+ requirement, glucagon has little (less than 2-fold) effect on the GTP[gamma S] requirement of N activation. These experiments indicate that the glucagon receptor facilitates activation of N by: (i) decreasing the apparent Km of N for Mg2+, and (ii) increasing the extent of activation that can be elicited by saturating concentrations of guanine nucleotide. It is postulated that the mechanism by which Mg2+ and receptors facilitate N activation involves dissociation of n alpha activated ADP ribosylatable subunits (with guanine nucleotide bound to them) from n beta non ADP-ribosylatable subunits (with receptor and Mg2+ bound to them). PMID- 6291030 TI - A prokaryotic membrane anchor sequence: carboxyl terminus of bacteriophage f1 gene III protein retains it in the membrane. AB - Gene III protein of bacteriophage f1 is inserted into the host cell membrane where it is assembled into phage particles. A truncated form of gene III protein, encoded by a recombinant plasmid and lacking the carboxyl terminus, does not remain in the membrane but instead appears to slip through it. Fusion of a hydrophobic "membrane anchor" from another membrane protein, the gene VIII protein, to the truncated gene III protein (by manipulation of the recombinant plasmid) restores membrane anchoring. A model for the relationship of gene III protein with the Escherichia coli membrane is discussed. PMID- 6291029 TI - Fragments of the simian virus 40 transforming gene facilitate transformation of rat embryo cells. AB - Segments of the simian virus 40 (SV40) genome that encode only fragments of large tumor antigen can facilitate immortalization of secondary rat embryo cells. The phenotypes of the immortalized cells range from nearly "normal" to fully transformed. All of the cell lines contain SV40 sequences and express unstable NH2-terminal fragments of large tumor antigen. SV40 small tumor antigen does not appear to be essential for either immortalization or transformation. PMID- 6291031 TI - Molecular cloning and chromosomal mapping of a human locus related to the transforming gene of Moloney murine sarcoma virus. AB - Human DNA was analyzed for the presence of sequences homologous to the transforming gene (v-mos) of Moloney murine sarcoma virus. A single 2.5-kilobase pair (kbp) EcoRI-generated fragment of human DNA was identified by using cloned v mos as probe. This DNA was molecularly cloned in a bacteriophage vector. By heteroduplex and restriction enzyme analyses, this human DNA fragment, designated c-mos (human), contained a 0.65-kbp region of continuous homology with v-mos and was present as a single copy in human DNA. By testing for the presence of c-mos (human) in somatic cell hybrids possessing various numbers of human chromosomes, as well as in subclones of such hybrids, it was possible to assign c-mos (human) to human chromosome 8. PMID- 6291032 TI - In vitro transcription of the thymidine kinase gene of herpes simplex virus. AB - We transcribed in vitro a cloned 3.5-kilobase fragment of herpes simplex virus type 1 DNA that contains the gene for the viral thymidine kinase. Extracts from uninfected HeLa cells produced five in vitro transcripts, one of which initiated at the in vivo start site for the thymidine kinase mRNA (an early viral message). A second in vitro transcript initiated at or near the start site for a major late in vivo viral mRNA. The remaining three in vitro transcripts may correspond to minor in vivo mRNA species. Sequences similar to the "T-A-T-A" and "C-A-A-T" boxes, which may be involved in the control of transcription of a variety of viral and cellular genes, were found to precede the initiation site of each of the five in vitro transcripts. Considerable overlap of transcription units was observed. PMID- 6291033 TI - Sarcomeric myosin heavy chain is coded by a highly conserved multigene family. AB - pMHC25, a recombinant plasmid containing myosin heavy chain (MHC) cDNA sequences from differentiated myotubes of the L6E9 rat cell line, has been shown to hybridize to all sarcomeric MHC mRNAs so far tested but not to nonsarcomeric MHC mRNAs. In addition, pMHC25 hybridizes to multiple restriction endonuclease digested fragments of rat genomic DNA corresponding to different MHC genomic sequences. Thus, the MHC gene represented by pMHC25 is a member of a sarcomeric MHC multigene family that has regions of sequence homology shared among its members. This sarcomeric MHC multigene family has been estimated to be composed of a minimum of seven genes, some of which are polymorphic in the rat. We have also determined that pMHC25 hybridizes to MHC gene sequences in genomic DNA of all species that have striated muscle, ranging from nematodes to man. Sarcomeric MHC genes, therefore, have been horizontally and vertically conserved in evolution. Additionally, we have used the pMHC25 plasmid to demonstrate that MHC genes do not undergo rearrangement or amplification during muscle cell differentiation. PMID- 6291034 TI - Sequence studies of several alphavirus genomic RNAs in the region containing the start of the subgenomic RNA. AB - The alphaviruses produce two mRNAs after infection: the genomic (49S) RNA which is translated into the nonstructural (replicase) proteins and the subgenomic (26S) RNA which serves as the mRNA for the virion structural proteins. The sequence of the region of the genomic RNA that contains the 5' end of the subgenomic RNA and the 5' flanking sequences in the genomic RNA were determined for several alphaviruses. A highly conserved sequence of 21 nucleotides was found which includes the first two nucleotides of the subgenomic RNA and the 19 nucleotides preceding it. We propose that the complement of this sequence in the minus strand is the recognition site used by the viral transcriptase for initiation of transcription of 26S RNA and that, in general, such short recognition sequences are commonly used among the RNA viruses. The COOH-terminal sequence of the nonstructural polyprotein precursor has been deduced for each virus. These protein sequences are highly homologous and are followed by multiple in-phase termination codons clustered in the nontranslated region of the 26S RNA in each case. In contrast to the proposed transcriptase recognition site, the particular triplets used for a given conserved amino acid have diverged markedly during evolution of these viruses. The protein homology is sufficient, however, for deduction of the correct coding phase of the RNA and allows the alignment of the corresponding nucleic acid sequence data from different alphaviruses without knowledge of the sequence of the entire genomes. PMID- 6291035 TI - Rapid and transient localization of the leader RNA of vesicular stomatitis virus in the nuclei of infected cells. AB - The leader RNA transcript from the 3' end of the genome of vesicular stomatitis virus (VSV) has been detected in both the nucleus and cytoplasm of infected baby hamster kidney (BHK) cells. In the cytoplasm, leader RNA accumulated gradually throughout the infection to about 200 molecules per cell at 6 hr after infection. In the nucleus, however, there was a sharp and rapid increase in the concentration of leader RNA to approximately equal to 300 molecules per cell at about 2 hr after infection that decreased rapidly by 3 hr. This report presents evidence for nuclear localization of transcription products of a (-)-strand RNA virus other than influenza and supports the hypothesis that the leader RNA plays a role in the shutoff of host cell transcription. PMID- 6291036 TI - Electron microscopic study of natural killer cell-tumor cell conjugates. AB - Natural killer (NK) cells were obtained from C3H mouse spleens according to a modified version of the method of Kuribayashi et al. [Kuribayashi, K., Gillis, S., Dern, D. E. & Henney, C. S. (1981) J. Immunol. 126, 2321-2327]. These cells retain in vitro cytotoxicity against certain model tumor cell targets and appear homogeneous by morphological criteria. NK cells, YAC (tumor) cells, and NK cell YAC cell conjugates have been examined with scanning (SEM) and transmission (TEM) electron microscopy. SEM experiments have shown that: (i) NK cells are large and possess various shapes in contrast to the YAC target cells which are smaller and round, (ii) YAC cells have uniformly distributed microvilli whereas the NK cell microvilli are most prominent in the area of effector-to-target contact, and (iii) in the absence of target cells, NK cell microvilli are found in a small number (usually 1-3) of cell surface locations. The region of NK cell-tumor cell contact has also been examined with TEM. The cells were stained with ruthenium red/OsO(4). The electron-dense ruthenium red/OsO(4) reaction product was consistently found in regions of close cell-cell contact, suggesting that carbohydrates were not completely cleared from areas of contact and that target and effector membranes do not fuse extensively. TEM observations indicate that NK cells have structurally unique granules. The granules are composed of at least two distinct compartments. The outer compartment contains the lysosome-associated enzymes acid phosphatase and inorganic trimetaphosphatase. No enzymatic activities have been found associated with the inner compartment. NK cells appear to degranulate when incubated with YAC cells. Under those circumstances, limited areas of the NK cytoplasm contain vacuole-like areas possessing granules and apparent granular debris. Degranulation appears to be involved in the cytotoxic function of NK cells. PMID- 6291037 TI - Immunofluorescent localization of the transforming protein of Rous sarcoma virus with antibodies against a synthetic src peptide. AB - Antisera were raised against a synthetic peptide (src-c) containing the six COOH terminal amino acids of p60src, the transforming protein of Rous sarcoma virus (RSV). Antibodies specific for the src-c peptide were purified by affinity chromatography and then used to study the location of p60src in transformed cells. The distribution of p60src was compared to that of vinculin, a candidate cytoskeletal substrate of p60src, by indirect double immunofluorescence microscopy. In RSV-transformed rat, mouse, and chicken cells, an extensive codistribution of p60src with vinculin was observed. Both proteins were concentrated in the few remaining focal adhesion plaques, in transformation induced rosette clusters at the ventral cell surface, and in cell-cell contact areas. In addition, antibodies to both proteins stained the cytoplasm diffusely. In all cells examined, the immunofluorescent staining patterns produced by antibodies to the src-c peptide were indistinguishable from those obtained by immunolabeling of p60src with sera from RSV-infected tumor-bearing rabbits. The excellent agreement of the results obtained with two completely independent antibody preparations indicates strongly that the observed immunolabeling patterns correctly define the intracellular distribution of p60src. The significance of the intracellular location of p60src to the transforming activities of the protein is discussed. PMID- 6291038 TI - DNA methylation in chicken alpha-globin gene expression. AB - We have investigated certain specific methylation sites of the chicken alpha globin gene cluster in DNA from embryonic and adult erythroid cells as well as from brain and sperm cells. Eight contiguous DNA fragments of the alpha-globin gene cluster were subcloned from a recombinant lambda phage. The subclones were used as probes to map all the Msp I/Hpa II and Hha I sites in the unmethylated cloned DNA and specific sites of methylation in and around the alpha-globin gene cluster in chromosomal DNA. The data show that sperm DNA is totally methylated at these restriction sites in the globin gene region, as is brain DNA, with some exceptions. Interestingly, the methylation status of specific sites 5' to the coding sequences is correlated with expression of the embryonic or adult alpha globin genes in different stages of erythroid development. Some sites showing partial methylation, however, do not conform to the model that transcribed genes are unmethylated or undermethylated. We also find a well-defined 3.5-kilobase region of DNA 5' to the alpha-globin gene cluster in which all C-C-G-G sites are resistant to Msp I digestion in all tissues. This "Msp block" is presumably caused by 5-MeCpC methylation. PMID- 6291039 TI - Tandem gene amplification mediates copper resistance in yeast. AB - Resistance to copper's toxicity in yeast is controlled by the CUP1r locus. This gene was cloned by transforming sensitive recipients (cup1(8)) with a collection of hybrid DNA molecules, consisting of random yeast DNA fragments inserted into the vector YRp7. Four resistant transformants were studied in detail. Autonomously replicating or integrated by homologous recombination into chromosomal sites, the corresponding plasmids and several subclones confer resistance on sensitive recipients carrying the natural variant allele, cup1(8). Tetrad analysis and genetic mapping established that integration occurs typically at the cup1(8) site located 28 centimorgans distal to thr1, a chromosome VIII marker. Restriction endonuclease cleavage and electrophoretic mobility studies revealed that the CUP1r locus consists of a tandem array of repetitive units. Each unit is 1.95 kilobases in length and contains single sites for Kpn I and Xba I and two Sau3A sites. The sensitive allele represents one repeat and the resistant allele embraces 15 tandemly arrayed repeat units. Progressive selections in higher copper concentrations establish strains with markedly enhanced resistance. Resistance, we propose, is mediated by a gene amplification mechanism based on unequal sister chromatid exchange. PMID- 6291040 TI - Incompatibility of plasmids containing the replication origin of the Escherichia coli chromosome. AB - Plasmids containing the replication origin of the Escherichia coli chromosome (oriC plasmids) are unstable in certain recA strains of E. coli. However, they can be maintained more stably in other recA strains. This stable maintenance has allowed us to study the incompatibility properties of oriC plasmids. We have found that two oriC plasmids are incompatible: they cannot be stably coinherited in individual dividing cells. An oriC plasmid is excluded from growing bacteria at a much faster rate in the presence of a hybrid plasmid made from an oriC plasmid and a high-copy-number vector plasmid than in the presence of another oriC plasmid. By inserting various segments around the oriC region into high-copy number vectors, we have shown that two different regions in the vicinity of the oriC region determine incompatibility. One region, which we named incA, includes the region essential for autonomous replication of the oriC plasmid. The other, incB, is adjacent to incA but is not required for autonomous replication. PMID- 6291042 TI - Genetic instability in Drosophila melanogaster: deletion induction by insertion sequences. AB - Females of Drosophila melanogaster heteroallelic or homoallelic for X chromosome giant (gt) mutants generate deletions involving the wild-type alleles at two X chromosome gene loci: yellow body color (y) and white eye color (w). The deletions, bidirectional in the case of y and with fixed endpoint in the case of w, are associated with particular X chromosomes. Distinctive insertion sequences, located proximal to the target loci, are presumed to generate the deletions. PMID- 6291041 TI - Single-copy sequence hybridizes to polymorphic and homologous loci on human X and Y chromosomes. AB - Use of a 4.5-kilobase-pair (kb) segment of single-copy DNA from a human genomic library as a hybridization probe of genomic human DNAs revealed allelic Taq I restriction fragments 10.6, 11.8, and 14.6 kb long. Among 12 unrelated individuals, all 6 males exhibited the 14.6-kb fragment in addition to one of the other fragments. Three of the females displayed 10.6- and 11.8-kb fragments, and the other three displayed only one fragment length; none had the 14.6-kb fragment. Hybridization of this probe to Taq I-digested DNAs from human-rodent hybrid cell lines (which have partial complements of human chromosomes) demonstrated segregation of the 14.6-kb fragment with the human Y chromosome and segregation of the 10.6- and 11.8-kb fragments with the human X chromosome. Furthermore, hybridization of this probe to Taq I-digested DNAs from 48 members of a single kindred revealed Y-linked inheritance of the 14.6-kb fragment and X linked inheritance of the 10.6- and 11.8-kb fragments. These experiments demonstrate homology between single-copy sequences on the human X and Y chromosomes. PMID- 6291043 TI - Viral etiology of juvenile- and adult-onset squamous papilloma of the larynx. AB - Juvenile- and adult-onset laryngeal papillomas were examined for the presence of a human papillomavirus (HPV) genome and capsid antigens. DNA was isolated from a portion of tissue removed for therapeutic purposes, and the presence of a papillomavirus genome was detected by Southern transfer analysis. The viral DNA found in the 12 juvenile-onset and the 8 adult-onset laryngeal papillomas examined was identified as HPV-6 on the basis of size, restriction endonuclease digestion patterns, and homology detected under stringent conditions. Restriction endonuclease analysis of the viral genomes revealed at least four different subtypes, designated HPV-6c through HPV-6f. The most common subtype, HPV-6c, was detected in over half of the papillomas studied, including both juvenile and adult types. The remaining tissue was fixed and processed for immunocytochemistry. The immunoperoxidase technique was used with an antiserum that reacts with capsid antigen(s) common to all HPV serotypes. HPV antigen was found in two of the juvenile-onset papillomas and two of the adult-onset papillomas. The antigen was localized to the nucleus and was distributed in the superficial layers of the epithelium. HPV capsid antigen had not previously been detected in cases of adult-onset papilloma, and the HPV genome in both juvenile- and adult-onset laryngeal papillomas had not been characterized. Despite the absence of detectable viral antigen in most of the specimens examined, the presence of the HPV genome provides strong evidence for the papillomavirus etiology of these tumors. PMID- 6291044 TI - Post-tetanic potentiation at an identified synapse in Aplysia is correlated with a Ca2+-activated K+ current in the presynaptic neuron: evidence for Ca2+ accumulation. AB - We have examined the presynaptic changes underlying post-tetanic potentiation (PTP) in Aplysia by using voltage-clamp techniques combined with specific pharmacological blocking agents. The amplitude and time course of PTP parallel a slow outward clamp current that we have identified as a Ca2+-activated K+ current. Because this current is proportional to intracellular Ca2+ concentration our findings provide evidence for the "residual Ca2+ hypothesis," according to which PTP is caused by the accumulation of intracellular Ca2+ after tetanus. To obtain further evidence for this mechanism we injected EGTA intracellularly and found that it decreased the duration of both PTP and the Ca2+ -activated K+ current. PMID- 6291045 TI - Map of distamycin, netropsin, and actinomycin binding sites on heterogeneous DNA: DNA cleavage-inhibition patterns with methidiumpropyl-EDTA.Fe(II). AB - We report a direct technique for determining the binding sites of small molecules on naturally occurring heterogeneous DNA. Methidiumpropyl-EDTA.Fe(II) [MPE.Fe(II) cleaves double helical DNA with low sequence specificity. Using a combination of MPE.Fe(II) cleavage of drug-protected DNA fragments and Maxam-Gilbert gel methods of sequence analysis, we have determined the preferred binding sites on a Rsa I EcoRI restriction fragment from pBR322 for the intercalator actinomycin D and the minor groove binders netropsin and distamycin A. Netropsin and distamycin A gave identical DNA cleavage-inhibition patterns and bound preferentially to A+T-rich regions with a minimal protected site of four base pairs. We were able to observe the effect of increasing concentration on site selection by netropsin and distamycin A. Actinomycin D afforded a completely different cleavage-inhibition pattern, with 4- to 16-base-pair-long protected regions centered around one or more G.C base pairs. PMID- 6291046 TI - Primary structure of the replication initiation protein of plasmid R6K. AB - The cistron of the replication initiation protein of plasmid R6K has been cloned into the single-strand DNA vectors M13mp8 and M13mp9 and its complete nucleotide sequence has been determined. The amino acid sequence of the initiator protein as predicted from its nucleotide sequence shows that the protein is lysine rich and weakly basic and has a molecular weight of 35,000, which is in close agreement with that estimated from the mobility in NaDodSO4/acrylamide gels. The secondary structure of the protein, approximately by the probabilistic methods of Chou and Fasman [Chou, P. & Fasman, G. (1978) Adv. Enzymol. 47, 45-148], suggests an NH2 terminal domain of primarily positively charged alpha-helical structure, a core region of interspersed short stretches of random coils and beta-sheets and turns, and a COOH-terminal domain of alpha-helix. PMID- 6291047 TI - Detection and cloning of human DNA sequences related to the mouse mammary tumor virus genome. AB - Sequences related to the mouse mammary tumor virus (MMTV) genome have been detected in fragments of restricted human cellular DNA. These results were obtained by using recombinant DNA containing the MMTV proviral genome and lowering the stringency of blot-hybridization conditions. The MMTV genome also reacts with unique families of fragments in restricted cellular DNA from other mammalian species but not with salmon sperm DNA. A clone that reacted with labeled MMTV proviral DNA was selected from a human DNA library in Charon 4A. Under stringent conditions, a 3.7-kilobase MMTV-related EcoRI fragment of this clone hybridized with many of the same EcoRI restriction fragments of human cellular DNA detectable with MMTV proviral DNA under low-stringency conditions. Specific fragments of the human clone were shown to contain sequences related to the molecularly cloned gag, pol, and env regions of the MMTV genome. PMID- 6291048 TI - Abelson murine leukemia virus: structural requirements for transforming gene function. AB - The integrated Abelson murine leukemia virus (A-MuLV) genome cloned in bacteriophage lambda gtWES.lambda B was used to localize viral genetic sequences required for transformation. Comparison of the biological activity of cloned A MuLV genomic and subgenomic fragments showed that subgenomic clones that lacked the 5' long terminal repeat and adjoining sequences (300 base pairs downstream of the repeat) were not biologically active. In contrast, subgenomic clones that lacked the 3' long terminal repeat and as much as 1.3 kilobase pairs of the A MuLV cell-derived abl gene were as efficient as wild-type viral DNA in transformation. The A-MuLV-encoded polyprotein P120 and its associated protein kinase activity were detected in transformants obtained by transfection with Cla I, BamHI, and HindIII subgenomic clones. In contrast, individual transformants obtained with subgenomic Sal I clones expressed A-MuLV proteins ranging in size from 82,000 to 95,000 daltons. Each demonstrated an associated protein kinase activity. These results provide direct genetic evidence that only the proximal 40% of abl with its associated 5' helper viral sequences is required for fibroblast transformation. PMID- 6291049 TI - Affinity labeling of the plasma membrane 3,3',5-triiodo-L-thyronine receptor in GH3 cells. AB - The binding of 3,3',5-triiodo-L-thyronine (T3) to GH3 rat pituitary tumor cells was studied at 15 degrees C and was shown to be saturable, reversible, and stereospecific. Least-squares analysis of the binding data showed two classes of binding sites with dissociation constants of 1.8 +/- 0.2 nM and 260 +/- 30 nM and binding capacities of (5.2 +/- 0.2) X 10(4) and (1.6 +/- 0.2) X 10(6) sites per cell, respectively. Affinity labeling of intact cells was carried out by incubation of cells with 0.3 nM N-bromoacetyl-[125I]T3 at 15 degrees C for 1 hr. Analysis of the cellular extracts by sodium dodecyl sulfate gel electrophoresis showed three labeled protein bands with apparent molecular masses of 55, 47, and 33 kilodaltons (kDal) in a ratio of 86:7:7. The labeling of only the 55-kDal protein band was selectively reduced to 50% by 20 microM unlabeled T3. Highly purified plasma membranes of GH3 cells were prepared and shown to be free of nuclei. Affinity labeling of the purified plasma membranes gave the same labeling pattern as with intact cells. Peptide mapping by Staphylococcus aureus V8 digestion of the 55-kDal protein from cells or plasma membranes gave the identical peptide fragments. Thus the 55-kDal protein labeled from intact cells is the same protein as that from purified plasma membranes. These results together with our earlier findings [Horiuchi, R., Cheng, S.-y., Willingham, M. & Pastan, I. (1982) J. Biol. Chem. 257, 3139-3144] suggest that the 55-kDal protein may be involved in mediating the uptake of T3 in GH3 cells. PMID- 6291050 TI - Microinjected progesterone reinitiates meiotic maturation of Xenopus laevis oocytes. AB - Microinjection of progesterone dissolved in paraffin oil induces the reinitiation of meiotic maturation in the Xenopus oocyte; 50% maturation is obtained when 50 nl of a 50 microM solution is microinjected into the oocyte. The kinetics of the response to microinjected progesterone are similar to the kinetics of response to externally applied hormone. When an aqueous solution of progesterone is microinjected instead of an oil solution, maturation is never observed, a result which confirms previous work. Leakage of the steroid into the external medium was estimated to range from 1.6 pmol/hr when microinjection was performed in oil to 3.6 pmol/hr when it was performed in aqueous solution. Metabolism of the hormone microinjected in oil is weak (less than 20%) as compared to that after aqueous microinjection (greater than 80%). Progesterone microinjected in oil decreases the cAMP content as does externally applied hormone. We therefore conclude that progesterone acts initially on an intracellular site in order to trigger meiotic maturation of the Xenopus oocyte. PMID- 6291051 TI - Frozen tissue sections as an experimental system to reveal specific binding sites for the regulatory subunit of type II cAMP-dependent protein kinase in neurons. AB - Specific binding sites for the regulatory subunit of type II cAMP-dependent protein kinase (RII) were revealed in neurons by an immunohistochemical approach. Fixed frozen sections of several regions of the rat central nervous system were incubated in the presence of bovine RII. Bound bovine RII was subsequently detected by an immunofluorescence procedure using antibodies that recognize bovine but not rat RII. The results indicate that RII binds with high affinity to neurons. Binding is prominent in dendrites and almost undetectable in axons and axon terminals. The morphological distribution of the RII binding sites is almost identical to that of microtubule-associated protein 2 (MAP 2) immunoreactivity. Preadsorption of RII with a MAP preparation highly enriched in MAP 2 completely abolished binding of RII to tissue sections, suggesting that the binding is mediated by MAP 2. Our results indicate that frozen sections of fixed tissues are a suitable experimental system for study of specific interactions of cellular macromolecules at a morphological level. PMID- 6291052 TI - Modulation of the epidermal growth factor receptor by platelet-derived growth factor and choleragen: effects on mitogenesis. AB - The addition of fresh medium supplemented with partially purified platelet derived growth factor (PDGF) to quiescent density-arrested cultures of BALB/c-3T3 cells decreases the subsequent binding of radiolabeled epidermal growth factor (EGF). The decrease in EGF binding can be observed 1 hr after the addition of PDGF. This effect is maximal in 2-3 hr, and binding remains diminished for at least 6 hr. These effects can be accounted for by a decrease in the number of EGF receptors with no change in receptor affinity. The action of PDGF is concentration dependent, but even at very high concentrations of PDGF the reduction in EGF binding is never more than 50%. Similar decreases in EGF binding are produced by other treatments that render BALB/c-3T3 cells competent, such as the addition of fibroblast growth factor or medium previously exposed to the macrophage-like cell line P388D(1). Cholera toxin (choleragen), which alone had no effect on EGF binding, dramatically potentiated the ability of PDGF to down regulate EGF receptors. Two to three hours after the addition of PDGF and choleragen, EGF binding was reduced by 80-90% compared with control values. The ability of PDGF and choleragen together to decrease EGF binding was substantially inhibited by cycloheximide. Autoradiography of [(3)H]thymidine-labeled cells shows that choleragen potentiates the action of PDGF; lower concentrations of PDGF are required to make cells competent after choleragen treatment. Furthermore, cells treated with PDGF and choleragen no longer require EGF for traverse of G(1) phase and initiation of DNA synthesis in defined medium. The reduction in receptor number produced by choleragen and PDGF, which may be due to internalization of the EGF receptor, may mimic the action of EGF and thereby remove the EGF requirement for DNA synthesis. PMID- 6291053 TI - DNA repair in Escherichia coli: identification of the uvrD gene product. AB - A 2.9-kilobase (kb) Pvu II DNA fragment that contains the uvrD gene of Escherichia coli K-12 has been cloned in both low-copy and multiple-copy plasmid vehicles. The low-copy uvrD plasmid (pVMK49) complements a variety of uvrD, uvrE, and recL mutations. In contrast, the same strains carrying the 2.9-kb fragment in a multiple-copy plasmid (pVMK45) remain sensitive to ultraviolet light (UV). Additionally, pVMK45 transformants of wild-type E. coli are sensitive to UV and methyl methanesulfonate and appear to be recombination deficient. The cloned uvrD gene does not complement the dominant uvrD3 allele. The 2.9-kb Pvu II insert in these plasmids encodes a single 76,000-dalton protein, which, on the basis of insertional inactivation experiments with the Tn1000 transposon, must be the uvrD gene product. These data confirm earlier genetic analysis which suggested that recL, uvrE, and uvrD were all allelic. The direction of transcription of the uvrD gene has also been determined. PMID- 6291054 TI - Movement of yeast transposable elements by gene conversion. AB - We have constructed yeast strains in which Ty (transposon yeast) elements at the HIS4 locus are genetically marked with the yeast URA3 gene. By isolating and analyzing Ura- derivatives of these strains, we have detected a variety of Ty mediated recombination events. In this paper, we describe events in which the DNA sequence of the Ty element at the HIS4 locus is replaced by the DNA sequence of a different Ty element. These replacements occur without alterations in the flanking DNA sequence and without chromosomal aberrations. We believe that these events result from gene conversion between the Ty element at HIS4 and a Ty element at a different site in the yeast genome. Gene conversion can occur between Ty elements that differ by large insertion and substitution mutations. These recombination events result not only in the movement of Ty sequences but also in alterations in expression of the adjacent HIS4 gene. Different Ty elements at the same site in the HIS4 regulatory region can result in His-, His+, and cold-sensitive His+ phenotypes. Several Ty elements render expression of the HIS4 gene subject to control by genes at the mating type locus. PMID- 6291055 TI - Herpesvirus-dependent amplification and inversion of cell-associated viral thymidine kinase gene flanked by viral a sequences and linked to an origin of viral DNA replication. AB - The genome of herpes simplex virus 1 or 2 consists of two components, L and S, which invert relative to each other during infection. As a result, viral DNA consists of four equimolar populations of molecules differing solely in the relative orientations of the L and S components. Previous studies have shown that the a sequences, located in the same orientation at the genomic termini and in inverted orientation at the L-S junction, play a key role in the inversion of L and S components. In this report we describe a virus-dependent system designed to allow identification of the viral genes capable of acting in trans to invert DNA flanked by inverted copies of a sequences. In this system, cells are converted to the thymidine kinase-positive phenotype with a chimeric plasmid carrying the thymidine kinase gene flanked by inverted copies of the a sequence and linked to an origin of viral DNA replication derived from the S component. The DNA introduced into the cells is retained and propagated in its original sequence arrangement as head-to-tail concatemers. Infection of these cells with herpes simplex virus 1 or 2 results in as much as 100-fold amplification of the plasmid sequences and inversion of the DNA flanked by copies of the a sequence. In infected cells, the amplified resident DNA accumulates in head-to-tail concatemers and no rearrangement other than the inversions could be detected. These results suggest that the a sequence-dependent inversions required trans acting viral gene products. PMID- 6291056 TI - Restriction map variation in the Adh region of Drosophila. AB - Restriction maps of a 12,000-nucleotide region containing the alcohol dehydrogenase (Adh) locus of the second chromosome of Drosophila were determined for (i) 18 independent second chromosome stocks of D. melanogaster established from several natural populations and (ii) five closely related Drosophila species. All of the detected map differences appear to lie outside the Adh coding block. Four polymorphic restriction sites and four insertion/deletion variants were identified among D. melanogaster chromosomes. The estimated heterozygosity per nucleotide site was 0.006. The relative distributions of restriction sites and insertion/deletion variations among subpopulations and species suggest that the insertion/deletion variation may be mildly deleterious. PMID- 6291057 TI - Human antibody to OFA-I, a tumor antigen, produced in vitro by Epstein-Barr virus transformed human B-lymphoid cell lines. AB - We established two long-term human B-lymphoblastoid cell lines (L55 and L72) transformed by Epstein-Barr virus that produced IgM kappa antibodies to the human tumor antigen, OFA-I. Periphral blood lymphocytes obtained from melanoma patients were used as the source of the B lymphocytes. Antibody specificity was determined by the immune adherence assay using various human cancer and noncancer tissues as targets. L55 antibody (designated anti-OFA-I-1) reacted with a variety of human tumor types whereas L72 antibody (designated anti-OFA-I-2) reacted only with tumor cells of neuroectodermal origin (melanoma, glioma, and neuroblastoma). The levels of IgM detected in the spent medium of 1 X 10(6) L55 and L72 cells were 4 and 9 micrograms/ml, respectively, by radioimmunoassay. PMID- 6291058 TI - Synthesis and secretion of alpha 2-plasmin inhibitor by established human liver cell lines. AB - The site of synthesis of alpha 2-plasmin inhibitor (alpha 2-PI), a physiologic inhibitor of plasmin, is not known with certainty. We have studied the production and secretion of alpha 2-PI by three established human liver cell lines derived from hepatocellular carcinoma and hepatoblastoma (Hep G2, Hep 3B, and PLC/PRF/5). As measured by a specific radioimmunoassay, the titer of alpha 2-PI increased in the medium of Hep G2 and Hep 3B cells with time, but no significant amount of alpha 2-PI was found in the medium of PLC/PRF/5. There was no evidence for a significant intracellular pool of this protein. On immunodiffusion against anti alpha 2-PI serum, alpha 2-PI secreted by Hep G2 (G2 alpha 2-PI) formed a simple precipitin line of complete identity with the alpha 2-PI present in plasma (plasma alpha 2-PI). G2 alpha 2-PI behaved similarly to plasma alpha 2-PI in Sephadex G-150 gel filtration, sucrose density-gradient centrifugation, and crossed immunoelectrophoresis. G2 alpha 2-PI inhibited plasmin activity instantaneously in a functional assay and formed a complex with plasmin demonstrable by crossed immunoelectrophoresis. De novo synthesis of alpha 2-PI was shown by the presence of specific immunoprecipitable radioactivity in the medium after 5 hr of labeling of the cells with [35S]methionine. Analysis of the immunoprecipitates by NaDodSO4/polyacrylamide gel electrophoresis showed a single peak of radioactivity corresponding to Mr 68,000. These results indicate that the liver is a site of alpha 2-PI production. PMID- 6291059 TI - Stable expression in mouse cells of nuclear neoantigen after transfer of a 3.4 megadalton cloned fragment of Epstein-Barr virus DNA. AB - All cells that harbor the Epstein-Barr virus (EBV) genome contain a neoantigen in the nucleus (EBNA). By transfection we located a segment of the genome that encodes or induces an antigen serologically related to EBNA. The responsible genes are found in the 3.4-megaldalton BamHI fragment K of EBV DNA, specifically in the left 1.9 megadaltons represented by HindIII fragment I1. Mouse LTK- cells were cotransformed with recombinant plasmids, containing the herpes simplex virus thymidine kinase gene and either EcoRI fragment B or BamHI fragment of K of EBV DNA. The TK+ cells surviving in selective medium were cloned. About 50% of the clones expressed the neoantigen in every nucleus. These mouse cells were used as antigens in immunofluorescence tests. Antibody to the nuclear antigen was found in 30 human sera known to contain antibody to EBNA; it was not detected in 18 sera that did not have antibody to EBNA. Mouse cells expressing EBNA as the result of acquisition of cloned EBV DNA fragments should prove useful in the characterization of the structure of this antigen and as reagents for the diagnosis of EBV infections. PMID- 6291060 TI - Mapping of polypeptides encoded by the Epstein-Barr virus genome in productive infection. AB - Over 30 viral-specified polypeptides are translated in vitro from RNA of cells productively infected with Epstein-Barr virus (EBV). The polypeptides map to sites in EBV DNA by hybrid selection. Almost all of the polypeptides are reactive with EBV immune human serum. Several of the polypeptides are part of the early antigen complex. Two others are likely to be major structural components of the virus. Genes encoding persistent early and late polypeptides are intermixed through most of the EBV genome. PMID- 6291061 TI - Regulation of dopamine- and adenosine-dependent adenylate cyclase systems of chicken embryo retina cells in culture. AB - We have obtained evidence that receptor-stimulated adenylate cyclase activity [ATP pyrophosphate-lyase (cyclizing), EC 4.6.1.1] is regulated physiologically in both embryonic and mature neurons. In a series of experiments using cultured retina cells from chicken embryos, we found that dopamine-sensitive adenylate cyclase activity spontaneously desensitized as cultures differentiated. The cellular response to dopamine reached a maximum after 5 days in culture and then decreased to 40% during the next 5 days. This spontaneous desensitization appeared to be caused by functional dopaminergic transmission because it could be blocked by the dopamine antagonist haloperidol. The ability of added dopamine at 100 microM to cause near-complete desensitization is consistent with this conclusion. Pharmacologically induced desensitization required 31 hr for maximal effect and was half-maximal at 1-10 microM dopamine. Analogous desensitization of the adenosine-dependent adenylate cyclase system also was noted. When dopamine was removed from the medium of chronically treated cultures, cells resensitized to subsequent stimulation at a very slow rate. Resensitization likely depended on replacement of dopamine receptors because chronic dopamine treatment caused the disappearance of binding sites for the ligand [3H]spiroperidol. In a second series of experiments, using hatched animals, we found that similar regulation of dopamine receptor binding sites and activity could be elicited by manipulation of environmental light, a treatment thought to influence dopaminergic transmission. Retinas from animals in constant light had less specific [3H]spiroperidol binding (35 fmol/mg of protein) than did retinas from animals in constant darkness (66 fmol/mg of protein) and made less cAMP in response to added dopamine. Our results indicate that regulation of the dopamine receptor system begins early in development and continues to function in mature synapses. PMID- 6291062 TI - A rapid method for the measurement of prostaglandin receptors. AB - A method is presented which provides for a simple and rapid determination of prostaglandin receptors in large numbers of tissue samples. Binding of prostaglandin F2 alpha, to preparations of microsomal membranes obtained from the ovaries of PMSG/hCG-treated rats served as the model system for this study. After incubating the membranes with [3H]-PGF2 alpha +/- graded concentrations of PGF2 alpha for 90 min at 22 degrees C, dextran-coated charcoal was added to adsorb the free PGF2 alpha during a 10 minute period. Centrifugation of the mixture for 3 minutes atop a 2.2M sucrose cushion resulted in the charcoal pelleting to the bottom while the [3H]PGF2 alpha-membrane complex remained at the interface. There was no further significant diffusion of PGF2 alpha after centrifugation, enabling large numbers of samples to be handled within a short period of time. The binding data revealed both high and low affinity receptors: Kd = 5.8 nM and 77 nM with binding capacities of 360 and 4000 fmole/mg protein. Both PGE1 and PGF1 alpha partially displaced [3H]PGF2 alpha from the binding sites whereas PGA2 was without effect. Binding equilibrium was reached within 90--140 min. The results are compatible with those of investigators using other techniques. PMID- 6291063 TI - The effects of arachidonic acid, PGI2, and 6-keto-PGF1 alpha on cyclic nucleotide concentrations in a ventilated and artificially perfused isolated dog lung. AB - In 35 isolated dog lung preparations, the pulmonary circulation of the right lung was completely separated from that of the left so that 1 lung of each preparation could serve as a control. The lungs were ventilated with 14% O2, 6% CO2 and 80% N2 and the pulmonary circulations were perfused with a dextran-salt-bicarbonate solution containing theophylline. Samples of perfusate were assayed for cyclic AMP and cyclic GMP (radioimmunoassay). Infusions of arachidonic acid (n=6) and PGI2 (n=4) but not 6-keto-PGF1 alpha (n=3) into the pulmonary circulation led to increases in cyclic AMP compared to control. Cyclic GMP levels were unchanged by the various infusions. Indomethacin (n=4) and acetylsalicylic acid (n=4) (prostaglandin (PG) synthesis inhibitors), and tranylcypromine (n=4) (a PGI2 synthetase inhibitor), prevented the cyclic AMP increases from arachidonic acid. This prevention was not the result of interference with the ability of cells to produce or release cyclic AMP since indomethacin (n=3), acetylsalicylic acid (n=3), and tranylcypromine (n=4) did not prevent cyclic AMP increases from PGI2. We conclude that infusion of arachidonic acid into the canine lung elevated pulmonary cyclic AMP but not cyclic GMP; that part or all of this increase most likely resulted from conversion of arachidonic acid to products of PG synthesis, particularly PGI2; that infusion of PGI2 mimicked arachidonic acid in that pulmonary cyclic AMP but not cyclic GMP was elevated, and that 6-keto-PGF1 alpha, a metabolite of PGI2, is unlikely to account for the cyclic AMP increases in this study. PMID- 6291064 TI - Dose-related facilitation and inhibition of passive avoidance behavior by the ACTH 4-9 analog (ORG 2766). AB - The effects of "low" and "high" doses of the ACTH 4-9 analog (ORG 2766) were studied on passive avoidance behavior of rats compared to ACTH 4-10 and [D-Phe7] ACTH 4-10. All peptides increased avoidance latency in a dose-dependent manner. However, "high" doses of ORG 2766 (500 and 1000 ng/rat) inhibited passive avoidance retention. "High" amounts of ACTH 4-10 and [D-Phe7] ACTH 4-10 still facilitated passive avoidance behavior. "High" doses of ORG 2766 like "low" amounts of this peptide delayed extinction of active avoidance behavior. "High" doses of [D-Phe7] ACTH 4-10 like "low" amounts of this peptide facilitated extinction of active avoidance behavior. The substituted analog apparently carries a dual effect on passive avoidance behavior. PMID- 6291065 TI - Influence of hashish extract on the social behaviour of encountering male baboons (Papio c. anubis). AB - The effects of hashish extract (2 mg delta 9-THC/kg)on the social behaviour of encountering male baboons were tested by ethological methods. In the "approaching" male the drug reduced "approach" and the aggressive elements "hit ground", "brows-back" and "attack" but increased the frequency of "retreat". Social interactions were generally diminished. In the "retreating" male friendly social interactions as "lipsmack" and "touch-back/handle-genitals" were suppressed but the threatening elements "open-mouth" and "tooth-grind" were stimulated. "Retreat" was additionally more frequent. Non-social activities and locomotion were not affected in either of the males. Treating both subjects with hashish resulted in a reduction of "lipsmack", "approach", "fight" and "chase" in the approaching and "lipsmack", "touch-back/handle-genitals", "chase", "retreat" and "flee" in the retreating male. Social activities were generally reduced in both animals. Comparing the behavioural effects of hashish in male baboons to those described in other non-human primates, in rodents but also in man revealed analogous effects in all species. The drug generally impaired social interactions, induced social withdrawal and led to social isolation of the drugged subject. PMID- 6291066 TI - Enkephalin analogue effects in the amygdala central nucleus on conditioned heart rate. AB - The experiment was conducted to assess the effects of enkephalin analogue administration into the amygdala central nucleus on the acquisition of classically conditioned heart rate responding in rabbits. Bilateral injections of either D-Ala2, Met5-enkephalinamide (DALA) or D-Ala2, D-Leu5-enkephalin (DADL) were administered in a 1.0 microliter volume into the central nucleus immediately prior to the conditioning session. Administration of DALA significantly attenuated the acquisition of conditioned heart rate responding whereas groups which received comparable doses of DADL exhibited conditioned responding which did not differ from the vehicle-injected control group. The effect on conditioned responding produced by DALA administration was blocked by concurrent administration of the opiate antagonist naloxone. These results provide some support for the involvement of mu receptor activity within the central nucleus region of the amygdala in conditioning processes. PMID- 6291067 TI - Microstructural analysis of the involvement of beta-receptors in amphetamine anorexia. AB - Rats were trained to take food by pushing the door of the pellet dispenser in an operant chamber. Log survivor analysis of the inter-response time frequency distribution was used to determine whether or not an animal was eating, at any time during a thirty minute session. This information was used to compute eating time, eating rate, and the mean length of bouts of eating and gaps between eating bouts. Video-recordings confirmed that the method discriminated eating from not eating with an accuracy of approximately ninety percent. Amphetamine (0.5 mg/kg) significantly reduced total food intake and eating time, and increased gap length; propranolol (5 mg/kg) significantly increased eating time and bout length. Following propranolol pretreatment, amphetamine significantly reduced eating time and bout length but also significantly increased eating rate; as a result there was no significant decrease in total food intake. The possible mediation of these effects by beta-adrenergic and dopaminergic systems is discussed. PMID- 6291068 TI - Intraseptally injected opiate agents: effects on morphine-induced behaviour of cats. AB - Behavioural effects of intraseptally administered opiate agents were analyzed in cats pretreated with an intraperitoneal injection of morphine. In this way, it became possible to investigate (1) the involvement of septal opiate receptors in the behavioural response of cats to systemic administration of morphine, and (2) the pharmacological character of septal opiate receptors. The following results were obtained with intraseptal injections 15-16 min after intraperitoneal morphine: (1) naloxone decreased frequencies of head and limb movements, and (2) morphine was ineffective. The following results were obtained with intraseptal injections 40-41 min after intraperitoneal morphine: (1) beta-endorphin and, to a lesser extent, fentanyl increased frequencies of locomotor patterns, (2) morphine and Met-enkephalin were ineffective, (3) naloxone and naltrexone decreased frequencies of locomotor patterns in a dose-dependent way, (4) naloxone and naltrexone antagonized the effects of beta-endorphin and fentanyl, and (5) morphine did not attenuate the effect of naloxone. The intraseptal injections affected only the frequencies of the systemically evoked behaviour patterns; the nature of the behaviour patterns remained unchanged. It is concluded that (1) systemically administered morphine does not affect behaviour via a direct action on septal opiate receptors, and (2) the receptors mediating the septally evoked effects are most probably epsilon-type opiate receptors. The hypothesis is put forward that systemic administration of morphine results in an increased release of beta-endorphin from hypothalamo-septal neurons and, as a consequence, changes the beta-endorphin activity at the epsilon-type opiate receptors in the septum. PMID- 6291070 TI - Lack of behavioral evidence for dopamine autoreceptor subsensitivity after acute electroconvulsive shock. AB - A single electroconvulsive shock (ECS) delivered 7 days previously produced a significant decrease in spontaneous locomotion and rearing in non-habituated rats but did not alter the inhibition of these behaviors produced by low dose apomorphine administration. As such apomorphine effects are thought to be mediated by dopamine (DA) autoreceptors, these data are not consistent with previous reports that the ECS treatment produces a persistent subsensitivity of DA autoreceptors. PMID- 6291069 TI - Memory and postsynaptic cholinergic receptors in aging mice. AB - Significant retention deficits were observed on passive avoidance tasks (step down and step-through) in 15-, 20-, and 25-month-old male C57BL/6 mice compared with 4- and 8-month-old mice. In contrast, cholinergic muscarinic receptor binding ( [3H]quinuclidinyl benzilate) in cerebral cortex, striatum, hippocampus, and cerebellum in these same animals revealed no difference in this 4- to 25 month age range. In a separate comparison of 4- and 29-month-old female mice, [3H]QNB binding was significantly decreased in the older group in cerebral cortex, hippocampus, and striatum. Environmental enrichment, compared with an impoverished environment, significantly improved retention in mice on 24-hr step down performance but affected QNB binding only minimally (6-7% decrease of QNB binding in cerebral cortex and hippocampus). Benzodiazepine ( [3H]flunitrazepam) receptor binding was significantly (12-15%) decreased in 29-month-old mice compared with 4-month-old mice in the cerebral cortex, hippocampus, cerebellum, and brain stem. PMID- 6291071 TI - Effect of lithium upon desipramine enhanced shock-elicited fighting in rats. AB - Rats were tested for changes in shock-elicited fighting (SEF) following the chronic administration of saline (IP); lithium (Li+) (20 mEq./l tap water) + saline (IP); desipramine (DMI) (15 mg/kg, IP); and DMI + Li+ for 14 days. The repeated test trials indicated a significant decrease in SEF in Li+-saline group (p less than 0.05), a significant increase (p less than p.05) in the DMI group, but no difference in the DMI + Li+ group in comparison to saline controls. Combined treatment with DMI + Li+ significantly reduced (p less than 0.05) SEF in comprison to the DMI group. These results suggest that enhanced aggressivity resulting from chronic DMI administration and measured by SEF can be a useful behavioral model to study the action of lithium. PMID- 6291072 TI - Phenytoin induces cyclic-AMP accumulation and free fatty acids release in rat brown adipose tissue. PMID- 6291073 TI - Effects of 6-hydroxydopamine pretreatment on in vivo femoral arterial responses to vasodilator agents. AB - Reserpine has been previously shown to generally enhance femoral arterial vasodilator responses as indicated by increased slopes of regression lines relating decreases in resistance produced by injections of vasodilator agents to resistance prior to the injections. We have examined the same parameters in dogs pretreated with 6-hydroxydopamine. In contrast, 6-hydroxydopamine either decreased (nitroglycerin, adenosine) or had no effect (isoproterenol, acetylcholine) on femoral arterial vasodilator responses. In a second set of animals, reserpine was administered after pretreatment with 6-hydroxydopamine, and reserpine again tended to augment responses to vasodilator agents. These data suggest that reserpine's effects on vasodilator responses may be a direct effect on vascular smooth muscle and was not dependent on its effects on sympathetic innervation of the femoral bed. The decreased vasodilator responses to certain agents induced by 6-hydroxydopamine, in contrast, may have been attributable to loss of a presynaptic component of their mechanism(s) of vasodilator action. PMID- 6291074 TI - Structure-activity relationships of peptides derived from ACTH, beta-LPH and MSH with regard to avoidance behavior in rats. PMID- 6291075 TI - Ultrastructure and calcium exchange of the sarcolemma, sarcoplasmic reticulum and mitochondria of the myocardium. PMID- 6291076 TI - Activation of the glucocorticoid-receptor complex. PMID- 6291077 TI - The acetylcholine receptor at the neuromuscular junction. PMID- 6291078 TI - Interactions between the renin-angiotensin system and the brain. PMID- 6291079 TI - Platelet vasopressin receptors in bipolar affective illness. AB - Determinations of the affinity constants (KD) and number of binding sites (Bmax) for platelet vasopressin receptors were done in euthymic bipolar patients and normal volunteers. No significant differences were found between these groups. In addition, lithium treatment did not alter these receptor parameters in the bipolar group. PMID- 6291080 TI - Plasma ACTH and cortisol concentrations before and after dexamethasone. AB - Alteration in the hypothalamic-pituitary-adrenal (HPA) axis occurs in up to 50% of depressed patients and is demonstrated by the failure to suppress cortisol concentrations after dexamethasone administration. Evidence suggesting that these cortisol abnormalities reflect hypothalamic-pituitary dysfunction has been inconsistent. We administered the dexamethasone suppression test to 28 psychiatric inpatients, including 17 cortisol suppressors and 11 nonsuppressors. Adrenocorticotropic hormone (ACTH) concentrations at 8 a.m. pre- and postdexamethasone were significantly greater in cortisol nonsuppressors than in suppressors. Our data support the hypothesis that pituitary ACTH secretion is altered in depressed patients who have HPA axis abnormalities demonstrated by plasma cortisol measurements. PMID- 6291081 TI - Plasma vanadium concentration in manic-depressive illness. AB - 133 samples of plasma taken from 9 normal control and 8 manic-depressive subjects were analysed for vanadium by atomic absorption spectrometry. Mean plasma vanadium concentrations were 0 . 15 microM in normal control, 0 . 34 microM in manic and 0 . 28 microM in depressed subjects, and 0 . 23 microM in manic depressive subjects after recovery. The differences between normal subjects and manic and recovered subjects were statistically significant. Significant negative correlations were found between plasma vanadium concentration and the ratio of Na K-Mg ATPase to Mg-ATPase in 2 manic-depressive subjects, but not in normal subjects. The results suggest that vanadium may be a cause of the variations in Na-K-Mg ATPase and sodium pump activity which are associated with manic depressive illness. PMID- 6291083 TI - Aggressive behavior and benzodiazepine receptor changes in isolated male mice. PMID- 6291082 TI - Some biochemical findings during pregnancy and after delivery in relation to mood change. AB - Thirty-four women who had vaginal deliveries of live infants completed behavioural ratings and supplied blood and 24-hour urine samples on three occasions during pregnancy and on three occasions after delivery. Approximately one third of the women indicated a distinct upswing in mood between Days 2 and 4 following delivery. These "positive mood change' subjects showed changes in urinary cyclic AMP (adenosine 3'5' cyclic monophosphate), plasma cyclic AMP, whole blood cell cyclic AMP, whole blood cell ATP (adenosine triphosphate), haematocrit and urinary 11 OHCS (11-hydroxycortisol steroids) following delivery which were different from those observed in the rest of the subjects and comparable with the biochemical changes described during upswings in mood in short-cycle manic-depressive subjects. There was also an indication that the women showing this upswing in mood following delivery were distinct in some respects on both behavioural ratings and biochemical findings during pregnancy. PMID- 6291084 TI - Discriminative stimulus properties of naloxone. AB - Nonopioid-dependent pigeons were trained to discriminate IM injections of 30.0 mg/kg naloxone from water in the procedure in which 15 consecutive responses on one of two keys resulted in grain presentation. Naloxone-appropriate responding was maximal at doses of naloxone equal to and greater than the training dose. The onset of the naloxone discriminative cue was rapid (less than 5 min) and the duration of the cue was short (less than 60 min). Naltrexone (1.0-10.0 mg/kg). pentazocine (1.0-10.0 mg/kg), levallorphan (1.0-30.0 mg/kg), and nalorphine (3.0 30.0 mg/kg) produced dose-dependent increases in naloxone-appropriate responding, occasioning 100% naloxone-key selections. In contrast, cyclazocine, profadol, and diprenorphine, at doses up to those at which animals did not respond, produced only intermediate degrees of naloxone-appropriate responding. Morphine always produced selection of the water key. These results demonstrate that a pure opioid antagonist, such as naloxone, has discriminative stimulus effects in the nonopioid-dependent animals, that these stimulus effects are shared by certain other opioid antagonists, and that these effects are distinguishable from those produced by pure opioid agonists, such as morphine. PMID- 6291085 TI - Up- and down- regulation of central postsynaptic alpha 2 receptors reflected in the growth hormone response to clonidine in reserpine-pretreated rats. AB - The alpha-adrenergic mechanisms exert a stimulatory influence on the secretion of growth hormone (GH) in the rat. In the present study the alpha receptors involved in GH regulation were characterized with respect to subtype. It was also investigated whether the GH response to alpha receptor agonists can be utilized to assess change in the responsiveness of central alpha receptors. The experiments were performed on rats with implanted intra-aortic cannulae allowing frequent blood sampling from freely moving animals. Plasma GH was determined by radioimmunoassay. Reserpine (10 mg/kg) caused a suppression of the normal pulsatile secretory pattern of GH. The alpha receptor agonist clonidine (CLON) given to reserpine-pretreated animals induced a dose-dependent increase in plasma GH. The effect of CLON (0.2 mg/kg) was prevented by pretreatment with the alpha 2 receptor antagonist yohimbine (3 mg/kg), but not by the alpha 1 receptor antagonist phenoxybenzamine (10 mg/kg). Chronic pretreatment with CLON or imipramine, either of which can be expected to produce a reduced sensitivity of central alpha 2 receptors, resulted in reduced GH responses to CLON. On the other hand, chronic treatment with yohimbine, which should cause denervation supersensitivity of alpha 2 receptors, led to enhanced GH responses to CLON. The results indicate that GH release in the rat is stimulated by postsynaptic alpha 2 receptors. They also suggest that the GH response to CLON can be used as a valid in vivo model reflecting decreased, as well as increased responsiveness of this type of receptor. PMID- 6291086 TI - The effect of hemodialysis on tardive dyskinesia. PMID- 6291087 TI - [The biological action of dithiocarbamates. Changes in serum leucine aminopeptidase induced in vitro by sodium diethyldithiocarbamate]. AB - The Authors demonstrate that serum LAP activity in vitro is inhibited by NaDDTC. This may be related to the NaDDTC chelating-like action on the metallo-enzyme. The trend of the phenomenon follows an exponential pattern. The enzyme activity was completely restored after removal of NaDDTC from the medium by dialysis. The NaDDTC concentration able to inhibit the enzymatic activity in vitro was much higher than in vivo, in experimental animals. The Authors conclude pointing out the problem of a probable effect caused by repeated doses of dithiocarbamates more on enzyme synthesis than on metallic apoenzyme. PMID- 6291088 TI - [Report of experiences with high-dosage single upper and lower half-body irradiation. I. Tumor modification. II. Reaction of lung tissue]. PMID- 6291089 TI - [Report of experiences with high-dosage single upper and lower half-body irradiation. III. Results of hematologic and special enzyme tests]. PMID- 6291091 TI - [Roentgen diagnosis of soft tissue tumors]. PMID- 6291090 TI - [Single half-body irradiation in bronchial carcinoma--initial results of a clinically controlled study]. PMID- 6291092 TI - Therapeutic alternatives in the treatment of intrahepatic biliary obstruction. AB - Intrahepatic biliary obstruction was treated in 60 patients (49 with cholangiocarcinoma and 11 with sclerosing cholangitis) who were classified according to the upper limit of their obstruction (Group I, proximal common hepatic duct; Group 2, right and left main hepatic ducts; Group 3, intrahepatic bile ducts). Thirty-six patients underwent percutaneous transhepatic biliary drainage, and 14 underwent catheterization through a T-tube track, Five of this latter group had the T-tube placed to establish a route of access for later interventional radiologic manipulations. Since most diseases that produce intrahepatic biliary obstruction are progressive, the use of any single approach is limited. The advantages of a surgically created route of access combined with the flexibility of interventional radiologic techniques help to maximize the therapy and extend the palliation that many of these patients receive. PMID- 6291093 TI - Obstruction of the left hepatic duct: diagnosis and treatment by selective fine needle cholangiography and percutaneous biliary drainage. AB - Obstruction of the left hepatic duct due to periportal pathologic conditions was identified by fine-needle transhepatic cholangiography in 28 patients. Selective cholangiography of the left duct was performed in 30 instances and subsequent selective left-sided catheter decompression was carried out in 23 cases. Techniques for selective puncture and drainage of the left duct required procedure modifications to accommodate the specific anatomic and pathologic features of the obstructed left duct system. Ultrasonography was indispensable as an aid to delineation of the left duct anatomy for directing needle puncture. Accurate documentation and successful catheter drainage of left duct obstruction are important contributions to the total management of patients with high biliary obstruction. PMID- 6291094 TI - Scintigraphic diagnosis of tricuspid regurgitation. AB - The authors describe a simple technique for diagnosis of tricuspid regurgitation. Red blood cells were labeled in vivo with 99mTc and 22 patients were studied with ECG-gated blood-pool imaging of the liver. A single region of interest was manually drawn around the liver and a time-activity curve obtained. The per cent change in liver counts during the cardiac cycle was found to be significantly higher in the 12 patients with tricuspid regurgitation (Group I) (mean, 4.04 +/- 1.6%; range, 1.3-21.4%) compared with the 10 controls (Group II) (mean, 0.35 +/- 0.16%; range, 0.013-1.3%) (p less than 0.05). Using a 1% change in liver counts as the criterion of a positive study, all 12 cases in Group I were diagnosed correctly, but there was one false positive in Group II; thus the sensitivity was 100% and the specificity 90%. PMID- 6291096 TI - [Genomic double stranded DNA of plant viruses]. PMID- 6291095 TI - Work in progress: a comparison of data collection protocols for single-photon emission tomography: 180 degrees versus 360 degrees. PMID- 6291097 TI - Higher-order conditioning and sensory preconditioning of a taste aversion with an exteroceptive CSi. PMID- 6291098 TI - The effects of a flavour-toxicosis pairing upon long-delay, flavour aversion learning. PMID- 6291099 TI - Degradation of cholecystokinin octapeptide, related fragments and analogs by human and rat plasma in vitro. AB - The rate of degradation of cholecystokinin octapeptide, related fragments and analogs by human and rat plasma was investigated, using high pressure liquid chromatography for the separation and identification of the degradation products. CCK tetrapeptide showed a half-life of 13 min in human plasma while its cleavage in rat plasma occurred at a very high rate (half-life of less than 1 min). The kinetics of disappearance of both sulphated and unsulphated CCK-8 indicated more than a single rate of degradation; the faster degrading system showed a half-life of 18 min for unsulphated CCK-8 and of 50 min for the sulphated peptide in human plasma as compared respectively with 5 and 17 min in rat plasma. The cleavage of CCK-8 was inhibited by bestatin and puromycin, suggesting that aminopeptidases play a major role in the breakdown of this peptide. CCK-9 analogs were rapidly converted into their corresponding octapeptides (half-life of 2.7 min in human plasma). This conversion was inhibited by puromycin and bestatin, suggesting the participation of aminopeptidase(s) probably specific for basic amino acids. CCK decapeptide exhibited a greater stability than the nonapeptides (half-life of 30 and 45 min in human and rat plasma respectively) and also gave rise to CCK-8 as degradation product. This cleavage was not affected by aminopeptidase inhibitors but was decreased by aprotinin (Trasylol), suggesting that trypsin-like and/or kallikrein-like enzyme(s) were involved in the plasma metabolism of this peptide. PMID- 6291100 TI - Specific photoaffinity crosslinking of [125I]cholecystokinin to pancreatic plasma membranes. Evidence for a disulfide-linked Mr 76 000 peptide in cholecystokinin receptors. AB - In rat pancreatic plasma membranes, preincubated with [125I]cholecystokinin-33 (CCK-33) and washed free of unbound tracer, the irradiation by UV light induced the irreversible binding of radioactivity to high molecular weight peptides as shown by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate (SDS) and autoradiography. This was not observed when the membranes were preincubated in the simultaneous presence of [125I]CCK-33 and of either an excess of unlabelled CCK-8 or of guanosine 5'-(beta, gamma-imido)-triphosphate. The radioactivity was mostly crosslinked with a Mr 96,000 peptide and peptide species of Mr greater than 200,000, after SDS solubilization in the absence of beta mercaptoethanol. Peptide reduction with beta-mercaptoethanol converted the high molecular weight radioactive species into a Mr 76,000 peptide that contained as much as 65% of the radioactivity crosslinked. The Mr 76,000 peptide appears, therefore, to be a disulfide-linked constituent of rat pancreatic cholecystokinin receptors. PMID- 6291101 TI - Deposition of 241Am and 239+240Pu at Tokai, Ibaraki Prefecture. AB - In order to estimate the fallout amounts of transuranium elements, monthly deposits have been collected at Tokai, Ibaraki Prefecture, during the period from October 1978 to March 1980, and were analyzed both on 241Am and 239+240Pu also on major surface soil components, SiO2 and Al2O3. Major soil matrices contents in the deposits suggested that 5.1% of the observed 239+240Pu deposition was contributed by resuspension from ground deposit with wind-blow, on the other hand, much greater contribution of 10.3% of 241Am was by the resuspension. Fallout amounts exclude resuspension of 241Am and 239+240Pu throughout the period were estimated to be 11.0 and 212 mBq/m2 (0.30 and 5.73 pCi/m2) respectively and annual values were 10.0 and 166 mBq/m2 . y) (0.27 and 4.49 pCi/m2 . y), respectively in 1979. Activity ratios of 241Am/239+240Pu changed widely. The average value of 0.066 was calculated from total amounts of each nuclides. PMID- 6291103 TI - [Clinical, histological and scanning electron microscopy evaluation of the Prophy Jet in vivo and in vitro]. AB - Clinical, histologic and scanning electron microscopic study of the use of the Prophy-Jet. The Prophy-Jet (Cavitron-International) is used for the removal of gross stain and heavy plaque. In this study the plaque and stain removal capacities have been investigated in patients with a healthy and inflamed gingiva. The influence of the spray of cleaning powder (sodiumbicarbonate) on the gingiva has been studied in gingival biopsies of Beagle dog. With the use of a scanning electron microscope the dentine and cementum have been investigated after spraying during 30 seconds to 2 minutes. The clinical results show that the plaque and the stain are removed easily and significantly except for the deep interproximal regions where the handpiece tip cannot be used in a correct angulation to the tooth surface. All patients showed gingival desquamation which could last up to 3 days. After treatment several patients suffered from aphthous ulcers. The gingiva of the Prophy-Jet treated areas in the Beagle dog showed severe epithelial desquamation, even of the papillary connective tissue, which in certain areas, can lead to necrosis. A spray lasting for 30 seconds removed approximately 10 micrograms of dentine or cementum. From these results it can be concluded that the negative aspects of the use of the Prophy-Jet are very important. It should therefore be used with caution. PMID- 6291102 TI - Aspects of male reproductive pharmacology and toxicology. PMID- 6291104 TI - [Breast tumor in adolescence]. PMID- 6291105 TI - [Importance of the measurement of nephrogenic cAMP in primary hyperparathyroidism]. PMID- 6291106 TI - [Pulmonary large-cell carcinoma with multiple metastases in the small intestine]. PMID- 6291108 TI - Colorectal cancer. Aetiology. PMID- 6291107 TI - [Necrosis of the bone marrow (2 cases diagnosed during life)]. PMID- 6291109 TI - Effect of aminoglutethimide on the structure and function of rat adrenals. AB - The effects of aminoglutethimide (AG) on adrenal ultrastructure, cholesterol accumulation, and steroidogenesis were studied in rats. AG was capable of altering the fine structure of the adrenocortical cells. In rats treated with this drug, adrenal mitochondria were polymorphic and their cristae were elongated and frequently showed unusual arrangement. These organelles tended to surround the lipid droplets and were often found to be gigantic in size. AG was also able to block the cholesterol side chain cleavage (CSCC) enzyme system, cause an accumulation of cholesterol in the adrenal gland, and cause decrease in serum testosterone concentration. When AG was administered along with adrenal corticotropic hormone (ACTH), the effects were more apparent. However, there was no difference in serum testosterone concentration between animals treated with AG in the absence or presence of ACTH. ACTH can significantly increase in serum testosterone in castrated rats, suggesting that the low level of testosterone in operated animals causes a reflex rise in luteinizing hormone (LH) which interacts with ACTH to enhance the adrenal androgen secretion. PMID- 6291110 TI - Novel calcium channels used for lanthanide ion-induced adrenal catecholamine release. AB - Catecholamine release by La3+, Ce3+, Pr3+, or Nd3+ from isolated bovine adrenals requires the presence of calcium in the perfusing medium. Calcium dependent, La3+ induced adrenal catecholamine release is enhanced at low pH and only slightly inhibited by pretreatment with the calcium channel blocker, verapamil. Since calcium channels in adrenal medulla used by lanthanides are not effectively blocked by hydrogen ions or by verapamil, they must be different from the calcium channels normally opened by the physiological neuromediator, acetylcholine. PMID- 6291112 TI - Border disease in goats in Norway. PMID- 6291111 TI - Re-investigation of the presence of UDP-glucuronosyltransferase in rat liver mitochondria. AB - Specific activities of UDP-glucuronosyltransferase and glucose-6-phosphatase, a marker enzyme of endoplasmic reticulum, were measured in mitochondria and microsomes. In mitochondria the specific activity of UDP-glucuronosyltransferase represented only 7-11% of that found in microsomes, when measured in the presence of various aglycone substrates, including 4-nitrophenol, 4-methylumbelliferone, 1 naphthol, phenolphthalein, testosterone and 17 beta-estradiol. Similarly, the specific activity of glucose-6-phosphatase in mitochondrial preparations was about 80% of that found in microsomes. In conclusion, it seems that in rat liver the UDP-glucuronosyltransferase activity associated with mitochondrial fractions reflects the presence of membrane fragments derived from endoplasmic reticulum, rather than mitochondrial location of this enzyme. PMID- 6291113 TI - Hemorrhagic shock. PMID- 6291114 TI - Measurement of transcutaneous PCO2 in critically ill patients. AB - Transcutaneous carbon dioxide tensions (tcPCO2) measured with a heated electrode were compared with arterial carbon tensions (PaCO2) in 28 patients. Seventy-eight observations were made. At a skin electrode temperature of 44 degrees C the tcPCO2 was significantly higher than PaCO2 but changes in PCO2 detected by both measurements were closely correlated. The correlation coefficient between tcPCO2 and PaCO2 was 0.92 (P less than 0.001). The 90% response time of the electrode in vitro was less than 1 min, and in vivo stabilization of the recording occurred in less than 15 min. It was found that the transcutaneous PCO2 electrode can be employed usefully in intensive care monitoring of adult patients especially when weaning them from artificial ventilation. PMID- 6291115 TI - A closed-chest model for the study of electromechanical dissociation of the heart in dogs. AB - A closed chest model in mechanically ventilated dogs is presented for the study of electromechanical dissociation (EMD) after defibrillation for ventricular fibrillation (VF). In seven dogs VF was induced by internal application of an AC current in the right ventricle. Precordial compression was not performed. When VF was continued for either 30 s or 60 s defibrillation typically resulted in complete restoration of electrical and mechanical cardiac activity. Immediately after defibrillation, the mean arterial blood pressure, pulmonary artery pressure, right ventricular filling pressure and cardiac output were markedly elevated; these elevations were accompanied by a slight increase in lactate and a decline in pH of the arterial blood. When VF was continued for 120 s EMD was observed in each instance after defibrillation in association with a regular rhythm. After 5-20 min, the ventricular fibrillation reappeared. The constancy of EMD in this model provides an experimental basis for study of clinical options, by which EMD may be prevented and/or mechanical competence of the heart may be restored. PMID- 6291116 TI - Risk of pulmonary complications in surgical patients. AB - The pulmonary complications in a group of more than 20 000 surgical patients between 1971 and 1980 were examined, as well as the pulmonary state of 100 surgical patients who died within 10 days of operation. The factors related to the patient's constitutions, nature of the surgical disease, operation and anaesthesia. Equal and homogeneous groups were considered in respect of each factor which were analyzed retrospectively from 1980 to 1971. The mean frequency of pulmonary complications in the Department of General Surgery was 12%. The main risk factors of pulmonary complications were: the duration beyond 4 h of the operation (59%), age over 70 years (48%), obesity (35%). They were followed in decreasing order by sepsis, hypovolemia and particular sites of operation. Severe pulmonary complications, bronchopneumonia, pneumonia and thromboembolism contribute to mortality. PMID- 6291117 TI - Prognostic factors in fresh water immersion. AB - The records of 63 consecutive fresh water immersion patients seen from 1969 to 1978 were examined retrospectively to evaluate those factors most relevant to the patients' eventual outcome. Nineteen patients had hematuria on admission, all 19 had complicated courses or died as a result of their immersion. Nine of the 63 patients had plasma hemoglobin concentrations of 30 mg/dl or above. Five of the nine died as a result of their immersion. The remaining four developed Adult Respiratory Distress Syndrome but survived. All other parameters measured seemed to have little effect on the prognosis. An elevation of PCO2 on admission seemed difficult to evaluate but might in certain circumstances suggest an ominous prognosis. PMID- 6291118 TI - Pulmonary artery pressure versus pulmonary capillary wedge pressure and central venous pressure in shock. AB - Evidence is presented from 43 dogs and 30 patients that under conditions of severe hemorrhagic, traumatic or septic shock, there may be partial obstruction of the pulmonary microcirculation due to disseminated intravascular coagulation (DIC) particularly in the pulmonary venules. This may cause the left atrial pressure to fall and the pulmonary artery pressure to rise, in some cases drastically. Pulmonary edema may result. This dangerous rise in pulmonary artery pressure is not reflected by the wedged pulmonary artery catheter which will monitor only the status of the left heart. Central venous pressure (CVP) may remain within normal limits even after pulmonary artery pressure has risen to dangerous levels with the development of pulmonary edema. It is only with right ventricle failure against the high pulmonary pressure that CVP rises. It is concluded that pulmonary artery pressure measurements are very important in monitoring intravenous fluid administration in severe shock. Wedged pulmonary artery pressures monitor the left heart but may be misleading if taken alone. Central venous pressure gives a delayed response to fluid overload. PMID- 6291119 TI - The infusion of catecholamines in dogs under general anaesthesia with fentanyl. The effects on intrapulmonary shunt. AB - The pulmonary venous admixture, PaO2, and pulmonary and systemic haemodynamics were studied in six mongrel dogs during infusion of dobutamine (infusion rate 7.5 micrograms . kg-1 . min-1), dopamine (7.5 micrograms . kg-1 . min-1) and isoproterenol (0.1 microgram . kg-1 . min-1). Anaesthesia was performed by a single injection of Fentanyl (0.35 mg/kg). The carbon dioxide tension and body temperature were strictly maintained within limits. Only isoproterenol produced a significant change in pulmonary arterial pressure from an average of 1.2 +/- 0.4 kPa to 1.6 +/- 0.2 kPa (P less 0.05). There was no significant change in systemic haemodynamics with any of the three drugs. The use of catecholamines in dogs with healthy lungs does not induce any development in pulmonary venous admixture when haemodynamics are unchanged. Changes in these variables are dependent upon changes in pulmonary blood flow rather than being direct effects of the catecholamine. PMID- 6291120 TI - [Leukocyte collagenase in peripheral blood of patients with rheumatoid arthritis]. PMID- 6291121 TI - [Morphological methods in the examination of connective tissue]. PMID- 6291122 TI - [Treatment of sporotrichosis with ketoconazole]. PMID- 6291123 TI - Serum protease inhibitory capacity. (Recent knowledge on alpha 1-antitrypsin deficiency). AB - The molecular structure and the serum levels of alpha 1-antitrypsin, the major antiprotease of human serum, are controlled by a series of codominant alleles at a single chromosomal locus, known as the Pi(protease-inhibitor) locus. The congenital deficiency of this inhibitor is known to be associated with the development of lung emphysema in early adulthood and chronic liver disease in childhood. Less frequent associations have been reported, such as rheumatoid arthritis, membranoproliferative glomerulonephritis and mosaicism for sex chromosomes. The identification of several suballeles of the Pi system, which was accomplished by means of a refinement of the isoelectric focusing technique, has promoted research concerning their possible pathogenic implications. The studies so far performed have often led to contradictory results, but nevertheless they strongly ascribe the property of controlling the quantitative levels of alpha 1 antitrypsin to certain M subtypes. Intermediate M3 subtype has recently been associated with the development of chronic obstructive lung disease in adulthood. Should this finding be confirmed by further evidence, a new approach to the prevention of lung disease could be considered, given that 30% of the individuals are carriers of the M3 suballele. In Italy, the incidence of congenital deficiency of alpha 1-trypsin appears to be greater in the northern regions, where 15-20 out of every 100,000 individuals are affected by the severe (ZZ) form of the deficiency. PMID- 6291124 TI - Modulation of beta-adrenergic receptor by hydrocortisone in human polymorphonuclear leukocytes. AB - We have examined the influence of isoproterenol and hydrocortisone on the cycle AMP content and lysosomal enzyme release from highly purified human polymorphonuclear leukocytes. It was found that isoproterenol, although a potent stimulus for cAMP accumulation in human lymphocytes, had only marginal effects on both cAMP content of PMN and enzyme release. Since corticosteroids have been reported to increase the response to beta-agonists in various tissues, it was considered that some of the anti-inflammatory potential of corticosteroids might be related to enhancing the response of PMN to beta-agonists. The influence of hydrocortisone on the response to isoproterenol was accordingly examined. It was found that hydrocortisone did in fact augment the isoproterenol-induced increase in cAMP content. In contrast to the potentiation of cAMP accumulation caused by hydrocortisone and isoproterenol together, the two agents produced only an additional inhibition of enzyme release. Since the inhibition of lysosomal enzyme release from human PMN did not closely reflect the changes observed in the cAMP content, a possible compartmentalization of intracellular cAMP is suggested. PMID- 6291125 TI - Percoll density gradient centrifugation. (An improved method for the enrichment of lymphocyte subsets mediating different activities). AB - Lymphocyte fractions obtained by Percoll density gradient centrifugation were assayed for their cytotoxic activities both in NK and ADCC cytotoxic systems. Furthermore, the release of a mediator, platelet slowing factor from the same subsets, was evaluated. Different mitogens (phytohemagglutinin, concanavalin A and lipid A) have been used for this purpose. The maximum of cytotoxic activities was recovered in E+ cell-enriched fractions. On the other hand, lipid A was able to stimulate the PSF production from enriched B cells (fraction I); phytohemagglutinin induced the release of PSF from E+ fractions (II and III) and concanavalin A from fraction II only. Our results suggest that Percoll density gradient is a very reproducible method in obtaining lymphocyte subsets enriched in cells mediating either cytotoxic activities or lymphokine release. PMID- 6291126 TI - [3 alternatives to official medicine]. PMID- 6291128 TI - [X-ray mammography and sonography of the female breast: a comparison]. PMID- 6291127 TI - [So-called sporadic ulcero-mutilating acropathy: neuropsychological study of 10 cases. II]. AB - Ten patients complaining of Sporadic Ulcerating and Mutilating Acropathy (SUMA) of Bureau and Barriere, also known as alcoholic-acrodystrophic neuropathy, were given the Wechsler-Bellevue's intelligence test (form I) and the Rey's memory test for standard psychometric evaluation. Pathologic mental deterioration of an organic type was found in a very large number of cases: 7 out of 10. The psychical picture that has been recognised is of three kinds: I) homogeneous mental deterioration of verbal and performance skills without any important memory feebleness (4 cases); II) partial mental deterioration shown by memory impairment as a single defect (1 case); III) total deterioration: kind I and II together (2 cases). The SUMA organic brain syndrome has not distinctive features as regards to common chronic alcoholism, but one can believe that it reveals itself with a higher frequency. This is likely to be in connection with a high individual susceptibility to the noxious effect of alcohol (wine) intake. Cerebral and peripheral neuronal damage seems to be established with the same developing modality during the time. PMID- 6291129 TI - [The effect of pneumoxide on Na+-K+ stimulated ATPase activity in the brain of rats of various ages]. PMID- 6291130 TI - Phospholipase C from Bacillus cereus has sphingomyelinase activity. AB - Purified, electrophoretically homogeneous phospholipase C (PLC) preparations can be separated into two peaks by isoelectric focusing in sucrose gradients. The main peak has an isoelectric pH of 6.6-6.8 and contains two Zn2+ per molecule. The more acid peak (isoelectric pH about 6.2) contains about one Zn2+ per molecule and has a markedly reduced specific activity which can be raised by adding Zn2+. The purified enzyme has a low sphingomyelinase activity which coincides completely with the lecithinase activity in fractions from isoelectric focusing. The sphingomyelinase activity was greatly enhanced by substitution of Co2+ for Zn2+ but remained essentially unaltered when the levels of Ca2+ and Mg2+ were changed. These findings provide evidence that the sphingomyelinase activity is a true endogenous activity of PLC and not caused by contaminating sphingomyelinase. PMID- 6291131 TI - Injury to human cells in culture induced by low density lipoprotein: an effect independent of receptor binding and endocytotic uptake of low density lipoprotein. AB - Cultured human endothelial cells isolated from umbilical cord veins and erythrocytes obtained from healthy donors were injured when exposed to low density lipoprotein (LDL). A close relationship between the amount of 125I-LDL associated with the cell surface and the degree of cell injury was demonstrated. This association occurred before any morphological signs of cell injury were observed and before any substantial release of 51Cr into the medium could be measured. Subsequent endocytotic uptake and lysosomal degradation of LDL did not seem to be a prerequisite for the LDL-induced cell injury to occur. Human serum albumin had an inhibitory effect on the association of 125I-LDL with the cell surface and in parallel a lowering effect on the 51Cr release. PMID- 6291133 TI - Comparison of two methods for measurement of serum angiotensin-converting enzyme in sarcoidosis. AB - A commercially available radiochemical (RC) assay (Ventrex) and a spectrophotometric (SP) assay for serum angiotensin-converting enzyme (SACE) were compared in 80 adults (40 blood donors, 20 patients with and 20 patients without sarcoidosis), covering a wide range of enzyme activities. A good agreement and a linear correlation (P less than 0.001) between the assays were observed. SACE were 25+/-5 kU/l (SP assay) and 98+/-18 kU/l (RC assay) in blood donors, corresponding to a conversion factor of 3.9. The RC assay may have some theoretical advantages, but both methods seem to be equally suitable for use in the management of sarcoidosis. The choice may partly depend on the requisite technical equipment. PMID- 6291132 TI - Serum lipoprotein and lymphocyte LDL receptor studies in parents and children with heterozygous familial hypercholesterolaemia. AB - In certain cases heterozygous familial hypercholesterolaemia (FH) cannot be confidently diagnosed from elevated serum lipid and lipoprotein values alone because of overlap with normal values. In the present study, therefore, lymphocyte high-affinity LDL receptor degradation of 125I-LDL was measured in 36 subjects from nine FH families. In eight families there was a good agreement between elevated serum lipid and lipoprotein values and reduced 125I-LDL degradation. In the ninth family, however, LDL receptor activity was normal in three subjects from three generations with elevated serum lipids and lipoproteins. The data suggest a disorder of LDL metabolism other than FH. PMID- 6291134 TI - The influence of prednisone on serum angiotensin-converting enzyme activity in patients with and without sarcoidosis. AB - The influence of prednisone on S-angiotensin-converting enzyme (SACE) activity was examined by serial measurements in 20 non-sarcoidosis patients and 8 sarcoidosis patients on prednisone, compared with 26 patients and healthy controls not under treatment. All but the sarcoidosis patients had SACE within normal limits (12.0-36.8 kU/l). In the three groups initial or pretreatment SACE levels (mean +/- SD) were 20.1 +/- 5.7, 66.9 +/- 27.4 and 26.4 +/- 4.1 kU/l, respectively. Non-sarcoidosis patients on prednisone had a lower pretreatment SACE than untreated control persons, presumably due to the different diseases represented in the groups. During the observation period SACE was rather stable in untreated patients and also in the treated group as a whole. But in 12 patients with SACE greater than 18.2 kU/l (mean of reference series minus 1 SD) a significantly decreasing SACE was observed after 1 week of treatment, whereas SACE was unchanged in patients with lower pretreatment levels, suggesting that in these patients prednisone could not decrease any further a level which was already low. In sarcoidosis the elevated SACE declined, reaching a normal level after 4 weeks on prednisone. In the assay ACE was markedly inhibited by SQ 14,255 (Captopril), moderately by methylprednisolone in concentrations exceeding those generally used clinically, while no inhibition was seen using acetylsalicylic acid. PMID- 6291135 TI - Enzymatic sulphation of bile salts in man. Bile salt sulphotransferase activity in percutaneous liver biopsy specimens from patients with liver disease. AB - Bile salt sulphation in liver disease was investigated by measuring the bile salt sulphotransferase level in percutaneous liver biopsy specimens from 27 patients. The same magnitude of mean specific enzyme activity was found in patients with cholestatic and non-cholestatic liver disease. No significant difference of the mean bile salt sulphotransferase activity was found when patients with and without reduced liver function as evidence from the intravenous galactose tolerance test were compared. The present results indicate that induction of liver bile salt sulphotransferase does not occur to a significant extend in clinical conditions with cholestasis. PMID- 6291136 TI - Calcium and stimulus-secretion coupling in gastric fundic mucosa. Effect of inhibition of calcium transport by verapamil on gastric acid secretion in the isolated guinea pig fundic mucosa and in healthy subjects. AB - The effect on gastric acid secretion of blocking transmembrane Ca2+ influx into the parietal cells has been studied in the isolated guinea pig fundic mucosa and in healthy volunteers. Verapamil inhibited in a dose-related manner histamine stimulated acid secretion in the guinea pig mucosa, whereas stimulation with theophylline and dibutyryl cyclic-AMP was unaffected. The effect of verapamil (Isoptin, 2.0 mg/h) on acid secretion stimulated by increasing doses (50, 200, 500 ng/kg-h) of 15-leucine synthetic human gastric I was studied in seven healthy volunteers, alone and in combination with infusion of calcium gluconate (1.0 meq Ca2+/kg-h). Verapamil inhibited the acid response to the lowest dose of gastrin, resulting in a significant increase of D50 of 15-leucine synthetic human gastrin I. This effect was partly reversed by calcium infusion. It is concluded that one of the mechanisms by which extracellular calcium concentration influences acid secretion is by transmembrane influx of Ca2+ during stimulation. PMID- 6291137 TI - Physiology of muscarinic functions. AB - The general features and distributions of muscarinic and nicotinic receptors are compared. The unusual aspects of the transmission of cholinergic impulses to the muscarinic receptors of eccrine sweat glands are discussed, and evidence is presented for an intermediate adrenergic step. The modulating roles of muscarinic receptors in sympathetic ganglia are considered as well as possible sources of the catecholamine involved in producing the inhibitory potential. The responses to activation of muscarinic receptors in the central nervous system suggest that they may regulate the level of neuronal excitability. Recent directions of research on the function of muscarinic receptors is summarized. PMID- 6291138 TI - Comparison between ranitidine, cimetidine, pirenzepine and placebo in the short term treatment of duodenal ulcer. AB - The therapeutic efficacy of ranitidine, pirenzepine, cimetidine and placebo in the 28 day treatment of duodenal ulcer was evaluated through an open randomized study performed in 120 patients. At the end of treatment, ranitidine, pirenzepine and cimetidine demonstrated a significantly higher efficacy on ulcer healing as well as on symptom relief in comparison with placebo (P less than 0.05). Data regarding cimetidine and ranitidine failed to reveal significant differences: pirenzepine, on the contrary, when compared with both the H2 blockers employed showed a slower effect on symptom disappearance. As far as functional data are concerned, placebo administration did not induce any variation of secretory or gastrinemic behaviour: on the contrary, after the end of ranitidine and cimetidine treatment a significant decrease of BAO and of MAO were found, failing to reveal any serum gastrin variation. After pirenzepine therapy no differences in the acid secretory behaviour were seen, while a significant increase of fasting gastrinemia was observed. PMID- 6291139 TI - Molecular events following activation of muscarinic receptors: the role of inositol phospholipids. AB - Muscarinic cholinergic agonists can provoke a wide variety of physiological responses in target tissues. This can be achieved by mobilisation of calcium from intracellular and extracellular stores. The members of the family of Ca2+ mobilizing receptors to which the muscarinic receptor belongs share the ability to provoke a calcium-independent decrease in the concentration of inositol phospholipids. This response is closely coupled to agonist binding and is thus implicated in the mechanism whereby muscarinic and related receptors bring about elevated levels of Ca2+ in cells. Another widespread response to activation of muscarinic receptors is inhibition of adenylate cyclase possibly through direct interaction with a nucleotide regulatory protein. Circumstantial evidence and analogy with alpha 2-adrenergic receptors suggests that adenylate cyclase inhibition may be mediated by a different form of the muscarinic receptor to that involved in calcium mobilisation. PMID- 6291140 TI - Muscarinic control of gastrointestinal motility. PMID- 6291141 TI - Electron microscopic detection of periodate reactive complex carbohydrates in human T and B lymphocytes. AB - The periodic acid-thiocarbohydrazide-silver proteinate (PA-TCH-SP) technique, which has the same cytochemical significance as PAS staining of light microscopy, was undertaken to reveal the distribution of complex carbohydrates with vicinal glycols in human lymphocytes in several conditions at the ultra-structural level. The PA-TCH-SP method stained the clustered and the scattered cytoplasmic granules, Golgi apparatus and glycogen particles of lymphocytes. In the buffy coat, the lymphocytes with the clustered cytoplasmic granules contained less glycogen particles compared with the lymphocytes without the clustered cytoplasmic granules. The majority of T-lymphocytes separated from the venous blood possessed PA-TCH-SP positive clustered cytoplasmic granules, but glycogen particles were scanty or negligible, while B lymphocytes were rich with glycogen particles but had scanty clustered cytoplasmic granules. The T and B cell-derived cultured cell lines had a similar reactivity to the peripheral T and B cells. PMID- 6291142 TI - Demonstration of chlamydial inclusions in exfoliated cells. AB - Microscopic demonstration of chlamydial inclusions within cells offered the first laboratory procedure supporting the clinical diagnosis of chlamydial infection. The cytological test for the detection of chlamydial inclusions in genital tract infection, through not as sensitive as isolation in cell cultures, is still of diagnostic value. The present study discusses the collection of clinical specimens for microscopic examination, the preparation, fixation and staining of slides by Giemsa stain, iodine, and immunofluorescence. PMID- 6291143 TI - Relationship between clinical and electrophysiological findings and indicators of heavy exposure to 2,3,7,8-tetrachlorodibenzo-dioxin. AB - In this study the prevalence rate of peripheral neuropathy in a population living in an area polluted with 2,3,7,8-tetrachlorodibenzo-dioxin (dioxin-TCDD) was determined. Of the 723 subjects invited to the first screening in 1977, 470 (65%) attended. At the second screening in 1978, of the 710 invited subjects, 319 (45%) attended. Prevalence rate ratios for peripheral neuropathy and the associated 95% confidence limits were calculated for subgroups determined by the presence of (i) predisposing factors to neuropathy (alcoholism, diabetes, occupational exposure to neurotoxic agents, etc) or (ii) conditions thought to result from exposure to dioxin-TCDD such as chloracne or abnormal serum hepatic enzyme levels. The prevalence rate of peripheral neuropathy among those subjects with predisposing factors and among those with chloracne or abnormal serum hepatic enzyme levels was nearly three times greater than among those without these manifestations. The results derived from this study may be useful qualitative pointers for identifying subjects at risk in the neurological follow-up. PMID- 6291144 TI - Polycyclic aromatic hydrocarbons in the occupational environment: with special reference to benzo[a]pyrene measurements in Swedish industry. PMID- 6291145 TI - Isocyanates represent a serious health hazard. PMID- 6291146 TI - Malignant mesothelioma in Turkish immigrants residing in Sweden. AB - Mesothelioma is a rare neoplasm in the general population, but it is strongly associated with previous asbestos exposure. The endemic occurrence of this disease in two villages in central Turkey has raised the question of whether the inhalation of naturally occurring zeolite dust may also be a factor in the development of mesothelioma. During the past few years a large portion of the inhabitants of one of the two villages of concern has immigrated to Sweden. This report presents three cases of malignant pleural mesothelioma and the results of a chest radiographic survey among these immigrants. Mineralogical analysis of lung tissue specimens from two of the cases revealed the presence of both zeolite and asbestos minerals and therefore suggested a synergistic effect involving both types of minerals. The importance of close medical surveillance of this high-risk population is emphasized, as is the possibility that similar cases appear in other countries because of increased migration. PMID- 6291147 TI - [Clinical death-cap (Amanita phalloides) poisoning: prognostic factors and therapeutic measures. Analysis of 205 cases]. AB - 205 cases of clinical poisoning with the mushroom Amanita phalloides (death cap) in the period 1971 to 1980 have been studied retrospectively. The lethality was 22.4%. Age and latency between the ingestion of the mushrooms and the first clinical symptoms were of prognostic significance. The death rate was 51.3% in children below 10 years of age but only 16.5% in patients older than 10 years. The average latency period was 10.3 hours for the fatal cases and 12.6 hours for the surviving patients. Country, year, sex and time of hospitalization did not influence lethality. Prognostic relevance could also be attributed to the thromboplastin time (Quick). 84% of the patients with values below 10% died, while all patients with minimal values of more than 40% survived. The correlation with the outcome was weaker for the serum transaminases and nil for creatinine. The patients underwent on the average 8 therapeutic measures, but up to 20 therapies could be administered to the same patient. Eight of the 30 recorded treatments involved general support, 7 toxin elimination and the remaining 14 could be classified as pharmacotherapy. With the aid of multiple regression analysis taking into account age, latency period and the effects of all the other measures, penicillin and hyperbaric oxygenation were found to contribute independently to a higher survival rate. As compared to penicillin, the combination of penicillin with silybin was associated with still further increased survival. On the other hand, several measures, including exchange transfusion, thiocytic acid, sulfamethoxazole, plasma expanders, haemodialysis, treatment of the hemorrhagic diathesis and THAM/sodium bicarbonate were administered more often to patients who did not survive. For the remaining 20 therapeutic measures our analysis revealed neither a positive nor a negative correlation with the clinical outcome. PMID- 6291148 TI - DNA topoisomerases. PMID- 6291149 TI - Calcium in synaptic transmission. PMID- 6291150 TI - Chromosomal localization of the human homolog (c-sis) of the simian sarcoma virus onc gene. AB - Nonrandom chromosome rearrangements of chromosome 22 have been identified in different human malignancies. As a result of Southern blot hybridization of a c sis probe to DNA's from mouse-human somatic cell hybrids, the human homolog (c sis) of the transforming gene of simian sarcoma virus was assigned to chromosome 22. Hybrids between thymidine kinase-deficient mouse cells and human fibroblasts carrying a translocation of the region q11-qter of chromosome 22 to chromosome 17 were also analyzed. These studies demonstrate that the human c-sis gene is on region 22q11 greater than qter. PMID- 6291151 TI - Cotransmitters in the motor nerves of the guinea pig vas deferens: electrophysiological evidence. AB - The contractile response of the guinea pig vas deferens to motor nerve stimulation is biphasic. The first phase is antagonized by the specific adenosine triphosphate-receptor antagonist arylazido aminoproprionyl adenosine triphosphate (ANAPP3), and the second by the alpha-receptor antagonist prazosin. The underlying electrical event, the excitatory junction potential, is also blocked by ANAPP3, but not by prazosin. PMID- 6291152 TI - Role of serotonergic input in the regulation of the beta-adrenergic receptor coupled adenylate cyclase system. AB - The action of desipramine on the norepinephrine-sensitive adenylate cyclase system and the density of beta-adrenergic receptors in rat cortex was studied after selective lesioning of serotonergic neurons with 5,7-dihydroxytryptamine. In animals with lesions desipramine failed to reduce the density of beta adrenoceptors but decreased the response of adenosine 3',5'-monophosphate to isoproterenol and norepinephrine to the same degree as in animals without lesions. The results demonstrate a functional linkage between serotonergic and noradrenergic systems in the rat cortex, with beta-adrenergic receptors and neurohormonal sensitivity of the adenosine 3',5'-monophosphate-generating system being under separate regulatory control. PMID- 6291153 TI - The chemistry of vision. AB - The visual response is initiated by light reception and transduction into chemical and electrical energy in the outer-segment membranes of rod and cone cells. Recent research on the molecular events controlled by light has clarified the roles of some of the rod outer-segment biomolecules. These developments and the current unresolved questions are described. PMID- 6291154 TI - Epidemiology and some prognostic factors in osteosarcoma in Sweden. AB - Since 1958, there are about 80 cases of primary malignant bone tumours registered each year to the Central Cancer Registry. During the first 10 years, that is 1958 -1968, 242 cases of osteosarcoma were registered (28.8% of all bone tumours- 22.9% for chondrosarcoma) the incidence should be calculated at 2-3 cases per 1 million a year. The distribution according to age and sex is the same as found in most western countries. In a group of osteosarcoma patients, Dr Brostrom could study the longitudinal growth from birth until the age of diagnosis and relationship with tooth eruption. It was found that they were statistically identical for the patient and for the controls. An other study demonstrated no difference in the 5-year-survival rate between patients who had an amputation without prior biopsy and those who had a biopsy. PMID- 6291156 TI - [Adjuvant chemotherapy for osteogenic sarcoma: analysis of the present situation and of factors which have influenced therapeutic results]. AB - Although it has been used for more than ten years, the value of chemotherapy as an adjunct to surgery has not yet been firmly established. Uncertainties are related to the multiplicity of factors involved in disease progression, management, and evaluation of results. Therefore, it is important to know the results of ongoing or terminated therapeutic trials, in order to propose new guidelines for research and for improvement of therapeutic results. The purpose of this article is to analyse the published results from historical controls (USA and Europe), and the results obtained using chemotherapy in the most significant series reported, as well as a series of twenty-six cases treated by the GESO (Osteogenic Sarcoma Study Group) following two different programs. The results in this group are similar to results in classical historical controls and in therapeutic trials reported by other spanish centers, although they are worse than those recorded in our own historical controls retrospectively analysed. It seems that in our patients unfavorable prognostic factors are predominant so that our results are worse than those from other series. Therefore, new controlled trials are needed, with adequate grouping of cases, using a different preoperative chemotherapy regimen, and comparing results to those in control groups without further treatment. PMID- 6291155 TI - [Canine osteosarcoma as a model in comparative oncology]. AB - Osteogenic sarcomas constitute 80% of all bone tumors in dogs, i.e. approximately 4% of canine tumors. Canine and human osteosarcomas have many clinical and pathological features in common. Epidemiological studies have provided interesting data: larger canine breeds are subject to excess risk of primary bone sarcoma, probably as high as thirty times the risk in small dogs. The appendicular skeleton, namely the metaphyses of long bones, is a preferential initial site of the tumor, and this involvement increases with the weight class of the animals. These data suggest that body size, weight or growth rate may act as factors of bone sarcoma development. Therapeutic trials on canine osteosarcoma are ongoing and could be used as models for the human counterpart. PMID- 6291157 TI - [Adjuvant therapy of osteogenic osteosarcomas of the limb: results of the SO4 78 trial]. AB - Twenty-nine patients with osteogenic osteosarcoma of a limb underwent both pulmonary radiotherapy and chemotherapy immediately after treatment of the primary tumor, mainly by radical surgery or radiotherapy (80 grays/tumor). Chemotherapy consisted of alternate A and B cycles every four weeks and BCG scarifications between cycles. Cycle A combines vincristine, ameticine, methotrexate and folinic acid; cycle B consists of adriamycin, vincristine, imidazole carboxamide and cyclophosphamide. A 20 gray irradiation was delivered to the thorax between the first A and B cycles. 50% of patients had no local recurrence or metastases after three years. 70% are alive at three years. Toxicity of this protocol is mainly hematologic and bronchopulmonary (with one fatal infection). Alopecia and minor digestive toxicity were recorded in all patients. Severe cardiotoxicity was seen in only one case. Longer follow-up is needed to evaluate long-term toxicity, particularly bronchopulmonary side effects. PMID- 6291158 TI - [Management of pulmonary metastases from osteosarcomas]. AB - The management of pulmonary metastases from osteosarcomas rests at present on thoracic surgery combined with chemotherapy. Until the beginning of the seventies chemotherapy proved very disappointing. With adriamycin, methotrexate in high doses followed by folinic acid and, more recently, platinum cis-dichlorodiamine, remissions, which are usually partial, are obtained in a significant proportion of patients (at least 30%). The addition of less active agents such as vincristine, cyclophosphamide, actinomycin D and bleomycin is helpful. Among 31 patients with pulmonary metastases from treated osteosarcomas seen at the Gustave Roussy Institute, 18 underwent thoracic surgery as the first treatment; in 10, surgery was followed by chemotherapy with adriamycin, vincristine, methotrexate in high doses + folinic acid + cyclophosphamide. Five patients are in complete remission 27, 30, 49, 50 and 77 months after the surgical procedure. 12 patients were initially treated with a similar chemotherapeutic regimen; a subsequent thoracic surgical procedure was undertaken in two patients who died 18 and 30 months after the pulmonary metastases had appeared. In one patient, the metastases were treated by irradiation. These results are compared to previous reports in the medical literature. PMID- 6291159 TI - [Prospects for conservative excision procedures in osteosarcomas in 1981]. AB - On the tenth anniversary of the Bone Tumor Study Group (BTSG) the authors report on their overall experience of the surgical management of osteogenic osteosarcomas. Current data from the medical literature is analyzed. The authors believe that every time a resection-reconstruction procedure is possible it should be preferred to systematic amputation. This policy is warranted by the continuing advances in medical management, particularly in chemotherapy. However, when dealing with osteosarcomas, the therapist's fore-most concern is the danger to life rather than functional considerations. Attention is drawn to recent technical developments enabling total excision of vertebral and costovertebral osteosarcomas, that should lead to considerable improvement of the prognosis of such tumors which was hopeless until now. PMID- 6291161 TI - [Metoclopramide in hematology]. AB - Metoclopramide is very useful in hematology. This French drug has been continuously used for twenty years as a powerful inhibitor of nausea and vomiting induced by numerous antineoplastic associations. When administered intravenously in patients under sequential chemotherapy for leukemia or lymphoma, metoclopramide prevents digestive discomfort. After each infusion, the ingestion of solution or tablets prolongs the antiemetic action for several hours without any side-effects. Conversely, corticosteroid therapy is well tolerated in ulcerous and gastritic patients and tolerance is preserved in others when they are submitted for the first time to high doses of metoclopramide, because of the acceleration of gastric evacuation. PMID- 6291160 TI - [Septicemia due to Leptotrichia buccalis in an immuno-suppressed patient]. AB - The authors report a case of septicemia due to Leptotrichia buccalis in a patient with underlying lymphatic leukemia. It is the second case reported in the literature. This observation shows, once again, that bacteria known as "non virulent" can exhibit pathogenicity in immunosuppressed patients. PMID- 6291162 TI - [The antiestrogen effect]. AB - The effects of estradiol, which are requisite for the normal occurrence of events related to reproduction, are not restricted to that field. They play a part in the overall trophicity of the female organism. On the other hand, relative or absolute estrogen hyperactivity results in harmful clinical effects and in disastrous cellular consequences, i.e. initiation or promotion of genital carcinomas. After a review of the different steps of the synthesis and action of estradiol, the various compounds that are active on these steps are considered: antigonadotropes (norsteroid progestogens, androgens, danazol, prolactin, corticosteroids, LHRH), inhibitors of estrogen synthesis (aminogluthetimide and testolactone), compounds that increase catabolism (progestogens), and antagonists of the cellular effect of estradiol (tamoxifen, nafoxidin, clomifen, progesterone, progestogens and androgens). The putative or established site of action and the main aspects of therapeutic use are given for each substance. PMID- 6291163 TI - [Blood sugar control in diabetics with the insulin pump. Comparison with discontinuous injections]. AB - Eighteen insulin-dependent diabetic patients received insulin in a continuous subcutaneous infusion delivered by a portable pump. Short-term glycemic control was evaluated on the average glycemia (serum glucose was assayed seven times per 24 hours for three to five days), on the MAGE coefficient which gives an estimate of glycemic lability, and, in eight patients, on serum C-peptide concentrations. Results were compared to those recorded with two daily subcutaneous injections of a combination of regular and intermediate insulin. The average glycemia and the MAGE coefficient were 1.32 + 0.05 milligram (m + SEM) and 1 + 0.05 milligram respectively under continuous infusion, compared to 2.05 + 0.12 and 1.76 + 0.12 milligram respectively under insulin injections. Insulin requirements were significantly lower under continuous infusion (0.99 + 0.08 U/kg versus 1.35 + 0.11 U/kg). In three patients with brittle diabetes the use of the pump ensured rapid metabolic control. The advantages and drawbacks of portable insulin pumps are discussed in the light of this experience. PMID- 6291164 TI - [Meso-omental cystic lymphoma. Considerations apropos of 4 cases]. AB - Cystic meso-omental lymphangiomas are uncommon benign tumors. When symptoms arise, the correct diagnosis is only exceptionally considered. These tumors may be revealed by a variety of clinical pictures which, in fact, are not specific. Cystic meso-omental lymphangiomas are usually diagnosed in pediatric patients, although they may arise in adults. The four patients who are reported on are adults. Diagnosis was established upon histopathological examination of specimens removed during surgery or autopsy. Data from a review of the medical literature provides a basis for a clinical and above all pathological study of cystic meso omental lymphangiomas. Prognosis is usually favorable although it is dependent upon the conditions of surgical management. PMID- 6291166 TI - [Extraskeletal epidural Ewing sarcoma. 2 anatomoclinical cases with review of the literature]. PMID- 6291165 TI - [Treatment of inflammatory breast cancer. Controlled study of a combination therapy program]. AB - The authors report 27 cases of inflammatory carcinoma of the breast (T3-T4, M0) and propose: induction chemotherapy (adriamycin, vincristin, methotrexate) conventional local and regional radiotherapy, sometimes complementary surgical exeresis, and a complement of consolidation chemotherapy (cyclophosphamid, methotrexate, fluoro-uracil or adriamycin, vincristin, and methotrexate). In these forms, where the prognosis is very bad, this therapeutic regimen may improve considerably the median remission and survival rates. But important therapeutic progress remains. PMID- 6291167 TI - [Late anuria in rectosigmoid cancers]. AB - The authors discuss nine cases of anuria by local recurrence after abdominoperineal resection for cancer of the rectosigmoid, with particular emphasis on the frequency, diagnosis, prognosis, and the different methods of treating this type of anuria. The placing of a Gibbons catheter between the renal pelvis and the bladder seems to be the most suitable. PMID- 6291168 TI - [Autochthonous Plasmodium falciparum malaria and the malarial liver]. AB - When P. falciparum malaria is diagnosed in France in a patient who has not travelled to an endemic area, transmission by blood transfusion or direct contact with an infected person, and indigenous malaria should be considered. With reference to the observation reported in this paper, the manifestations of malarial hepatitis are recalled and their diagnostic significance is discussed. In P. falciparum malaria, malarial hepatitis indicates the development of a complete cycle including the pre-erythrocytic phase; this is strongly in support of indigenous malaria, and against malaria following blood transfusion or direct contact in which there is no pre-erythrocytic phase. PMID- 6291169 TI - [Still disease in adults]. AB - In 1971, less than one-hundred years after Still's publication (1897) concerning children, Bywaters described the adult-onset form of Still disease. Over one hundred cases of this disorder have been recorded since. In adults, Still disease affects mainly women, and the average age at onset is twenty-seven. The well known features demonstrated by clinical examination and complementary investigations are recalled. They include fever, arthritis, rash, lymphadenopathy, enlarged spleen, and serositis. Attention is drawn to hepatic and muscular involvement as well as to articular prognosis. Additional information is provided on questions which remain open to debate, particularly concerning management and therapeutic results. PMID- 6291170 TI - [Contribution of immunochemical methods to the study of human red-cell membrane proteins]. AB - Together with the classical technics of biochemical analysis, immunological methods have led to differentiate, characterize the main red-cell membrane proteins and to better understand their organization. Immunological methods were particularly involved in the study of: 1) the membrane skeletal proteins, specially spectrin which localization in the membrane, structure and functions have been specified; 2) the principal integral proteins, glycophorin and protein band 3, the transmembrane orientation of which has been corroborated by topographic immunological mapping of their cytoplasmic domain; 3) the anchor chain protein, linking the membrane skeleton to the transmembrane proteins. These methods could further help to a better understanding of the membrane structure. Two kinds of work can be considered using monoclonal antibodies provided by the hybridoma method: 1) the purification by immunoabdsorbent technics, of quantitatively minor membrane proteins, would allow to study their structure and functions; 2) an immunological mapping with monoclonal antibodies specific against each of the skeleton proteins, of membrane preparations observed in electronic microscopy, would permit to visualize the real architecture of these different proteins in the red-cell membrane. PMID- 6291171 TI - Etiologic factors in cervical neoplasia. PMID- 6291172 TI - [Dietary fiber and diseases of the digestive organs]. PMID- 6291173 TI - [Aftercare department (for convalescents) based in central district and city hospitals]. PMID- 6291174 TI - [Cystosarcoma phylloides]. PMID- 6291175 TI - Some effects of gamma-linolenic acid on cultured human oesophageal carcinoma cells. AB - Cells of two continuous human oesophageal carcinoma lines were treated by the addition of gamma-linolenic acid (GLA) at concentrations of 10-60 micrograms/ml culture medium. After 7 days of exposure to GLA, pronounced morphological changes became evident and culminated in cell death. GLA-mediated effects were time- and dose-dependent and varied slightly with the degree of histological differentiation of the lines tested. These findings may have clinical implications. PMID- 6291176 TI - The reversibility of cancer: evidence that malignancy in human hepatoma cells is gamma-linolenic acid deficiency-dependent. AB - A further critical test of Horrobin's hypothesis that malignancy in cells may be dependent on gamma-linolenic acid (GLA) deficiency, has revealed that GLA supplementation produces a highly significant reduction in the growth rate (up to 87%) of a cultured human hepatoma cell line, compared with the growth rate of untreated hepatoma cells. This supports our previous suggestion that this hypothesis requires urgent further investigation at all levels including trials in human cancer patients. PMID- 6291178 TI - On the mechanism of methadone-induced alcohol consumption in humans. PMID- 6291177 TI - The radiological investigation of hepatocellular carcinoma in children. AB - Malignant liver tumours are not unusual in infancy and childhood. Hepatocellular carcinoma is however much less common than hepatoblastoma. Four children with hepatocellular carcinoma, 2 Black and 2 White, have been seen at Baragwanath Hospital and the Johannesburg Hospital in the past 8 years. Three children were under 4 years of age, with no known predisposing cause; 1 child was 11 years old and had a history of tyrosinosis. The radiological investigation of liver tumours is discussed. PMID- 6291180 TI - Effect of ethanol administration and withdrawal on GABA receptor binding in rat cerebral cortex. AB - Sodium independent GABA receptor binding was measured in synaptosomes prepared from cerebral cortex of rats made ethanol dependent by three daily ethanol administrations. In rats sacrificed 1 hour after the last ethanol dose there was a lower number of low affinity binding sites and lower affinity of the high affinity binding than in controls. The decreased affinity was present only in rats who showed symptoms of ethanol withdrawal during the course of ethanol administration. In rats sacrificed during ethanol withdrawal the affinity of the high affinity binding was lower than in controls and other binding characteristics were unchanged. This decreased binding was normalized by repeated Triton X-100 incubations indicating involvement of an endogenous inhibitor in this ethanol effect. Acute ethanol administration did not change GABA receptor binding. PMID- 6291179 TI - Antagonism of tertiary butanol by ACTH4-10, naloxone and dexamethasone. AB - Male C57 mice were pretreated with naloxone (4 mg/kg), ACTH4-10 (30 micrograms/kg), or saline 15 minutes prior to an injection of tertiary butanol (1.5 g/kg). Both naloxone and ACTH4-10 lengthened induction time (time to lose righting reflex) and shortened the duration of sleep time. Dexamethasone (.6 mg/kg) pretreatment of mice injected with ethanol (3.75 g/kg) or tertiary butanol (1.5 g/kg) lengthened induction time, but did not alter sleep time. The data do not support a primary role for acetaldehyde or TIQ involvement in the acute depressant effects of alcohol, but do suggest that some of the observed effects may in part be mediated through an endogenous opioid system. PMID- 6291182 TI - Serum calcium, parathyroid hormone, and urinary cyclic adenosine monophosphate after parathyroidectomy. AB - In specific cases of primary hyperparathyroidism (HPT), an intraoperative measure of parathyroid function might aid the surgeon. Ideally this would permit the surgeons to recognize that sufficient parathyroid tissue had been removed to cure the patient, but that viable glandular tissue remains. This study was designed to evaluate the efficacy of urinary cyclic adenosine monophosphate (cAMP) concentrations as such a determinant. We studied serum calcium and parathyroid hormone concentrations and urinary cAMP levels in 11 control patients undergoing thyroid and non-neck operations and in 22 persons undergoing parathyroidectomy for primary HPT. The serum calcium and parathyroid hormone concentrations were normal in control patients and elevated in each person with primary HPT. Changes of these parameters after successful parathyroidectomy took too long either to occur or to be measured to be clinically useful intraoperatively. Basal urinary cAMP concentrations were normal in 10 of 11 control patients and remained so during their operations. Elevations of basal urinary cAMP levels were found in 78% of those with primary HPT. At 30 minutes after removal of all abnormal parathyroid tissue, urinary cAMP levels remained high in 41% of those in whom it as elevated in the basal state. However, by 60 minutes all urinary cAMP values were normal or low. Plasma cAMP values were normal in three of four patients with primary HPT and did not change within 90 minutes after operation despite the performance of a successful parathyroidectomy. As expected, urinary cAMP levels returned to normal in each of these individuals. Intraoperative changes of urinary cAMP levels do reflect changes in parathyroid status. However, because of the delay of 40 to 60 minutes before urinary cAMP normalizes after parathyroidectomy and because of the sophisticated technology necessary for rapid determination of this cyclic nucleotide, its present clinical applicability is limited. PMID- 6291181 TI - Mammographic and pathologic correlation of microcalcification in disease of the breast. AB - Fifty-five patients with isolated microcalcification clusters and no palpable tumor of the breast were studied. All patients underwent appropriate localization biopsies and had roentgenographic, specimen roentgenographic examination of histologic confirmation that the area of microcalcification was removed. Fifteen patients proved to have a malignant lesion at biopsy and in 40 patients, it proved to be benign. Of the proved cancers, ten were noninvasive. Fourteen of the patients with cancer were free of axillary metastases. The results of the follow up study, with a mean time of four years, revealed a 100 per cent survival rate and no recurrence of disease. Careful study of the microcalcification clusters, at preoperative mammographic examination, did not show significant differences between the malignant and benign group that could be helpful to the clinician, in predicting the nature of the lesion. Due to the reasonably high, 27 per cent, rate of malignant disease, we believe all patients with isolated microcalcification clusters at preoperative mammographic examination should have a localization biopsy. PMID- 6291183 TI - [2d tumors in patients with subleukemic myelosis]. PMID- 6291184 TI - [Effect of enteric sterilization on the quantity of fecal microorganisms, incidence of infectious complications and dynamics of the peripheral blood neutrophil count during cytostatic therapy of hemoblastosis patients]. PMID- 6291185 TI - [May-Hegglin anomaly]. PMID- 6291186 TI - [Biochemical-pharmacological aspects of opiate addiction]. PMID- 6291187 TI - [Neurophysiologic aspects of opium addiction]. PMID- 6291189 TI - Bronchoscopic and percutaneous aspiration biopsy in the diagnosis of bronchial carcinoma cell type. AB - The cell type of bronchial carcinoma predicted from the results of bronchial biopsy at fibreoptic or rigid bronchoscopy or of percutaneous aspiration lung biopsy was compared with the type determined by histological examination of specimens obtained by thoracotomy, biopsy of an extrapulmonary metastasis, or necropsy in 180 cases. The rates of agreement with the final diagnosis were 95.7% for bronchial biopsy through the fibreoptic bronchoscope and 86.5% through the rigid bronchoscope. For percutaneous biopsy, which was usually carried out on tumours inaccessible to the bronchoscope, the rate of agreement was 61%, significantly lower than by the other methods (p less than 0.001). The diagnosis of oat-cell carcinoma by any technique was very reliable. Bronchial biopsy was more reliable than was percutaneous biopsy in diagnosing squamous-cell carcinoma. With any technique the commonest error was the incorrect diagnosis of squamous cell carcinoma or adenocarcinoma as large-cell undifferentiated carcinoma. PMID- 6291190 TI - [Drug-induced neurologic disorders. Peripheral neuropathies. Ototoxicity. Disorders of neuromuscular transmission]. PMID- 6291188 TI - Benign sclerosing pneumocytoma of lung (sclerosing haemangioma). PMID- 6291191 TI - Quantitation of the inhibitory effect of fibrinogen and its degradation products on fibrin polymerization. AB - Anticlotting activities of fibrinogen and its plasmin degradation products- fragments X,Y and D--have been measured. On the molar basis fragments Y and D are found equally active whereas fragment X acts about 2 times stronger. We suggest that the inhibitory effect depends on a set of specific binding sites characteristic of domain D. As fragment X possesses two such sets its activity is twice as high as that of the single-set fragments Y and D. In spite of its two domains D fibrinogen inhibits clotting much weaker than fragment X. This is probably due to the presence in fibrinogen of large COOH-terminal sections of the two A infinity-chains interfering with the inhibitory effect. The possible role of these A infinity-chain sections in fibrinogen-fibrin system is discussed. PMID- 6291192 TI - Inhibitory mechanism of dipyridamole on platelet aggregation ex vivo. AB - The effects of dipyridamole on platelet aggregation ex vivo and in vitro and on platelet cyclic AMP phosphodiesterase (PDE) were studied, and the mechanism of the ex vivo effects was assessed. Both the ADP- and collagen-induced aggregations ex vivo were inhibited dose-responsively by oral administration of dipyridamole. Maximum dipyridamole levels in the plasma were reached at 30 min after the administration. The inhibitory effects of dipyridamole on platelet aggregation ex vivo reached a maximum at between 1 and 2 hrs. On the other hand, the ADP-induced aggregation in vitro and cyclic AMP PDE activity were not inhibited until after 10 min of incubation at a low concentration of dipyridamole. This mode of inhibition of platelet aggregation in vitro and of cyclic AMP PDE activity agreed with the mode of inhibition in the case of platelet aggregation ex vivo. It is suggested therefore that the ex vivo effects, observed with only a low dipyridamole concentration in the plasma, may be due primarily to inhibition by dipyridamole of the cyclic AMP PDE in platelets. PMID- 6291193 TI - [The effect of isolated rearing and frequency of vaccinations on the development of humoral immunity to Newcastle disease and infectious bronchitis]. PMID- 6291194 TI - Sensory irritation, pulmonary irritation, and acute lethality of a polymeric isocyanate and sensory irritation of 2,6-toleune diisocyanate. PMID- 6291195 TI - Regional rat brain content of adenosine 3',5'-cyclic monophosphate and guanosine 3',5'-cyclic monophosphate after acute and subacute treatment with ethanol. PMID- 6291196 TI - Lead inclusion bodies in the anterior horn cells and neurons of the substantia nigra in the adult rhesus monkey. PMID- 6291197 TI - Isolation and characterisation of toxic components from the venom of the common Indian sea snake (Enhydrina schistosa). AB - Three neurotoxic components, enhydrotoxins a, b and c, were isolated by a three step procedure from the venom of the sea snake Enhydrina schistosa, caught in the Arabian sea. The i.v. LD50 values in mice were 0.042, 0.045 and 0.052 mg/kg, respectively. All three toxins irreversibly blocked neuromuscular transmission. Abolition of responses to acetylcholine indicated a post-synaptic site of action of these toxins. The molecular weight and amino acid composition of enhydrotoxin a are similar to values earlier reported for Enhydrina toxins. PMID- 6291198 TI - Effect of leishmanial excreted factor on the activities of adenylate cyclase from hamster liver and Leishmania tropica. AB - The carbohydrate segment of the excreted factor of culture cells from L. tropica and L. donovani was shown to interact with the adenylate cyclase from hamster liver. At a concentration of 1 mg/ml the excreted factor inhibited the stimulated activity of the mammalian enzyme by about 50%, independent of the activation by glucagon, 5'-guanylylimidodiphosphate and forskolin, respectively. In contrast to the adenylate cyclase system from hamster liver the enzyme from L. tropica culture cells was not stimulated by addition of glucagon, 5' guanylylimidodiphosphate and forskolin. In addition, the activity of the adenylate cyclase from L. tropica was not affected by addition of its endogenous excreted factor. PMID- 6291199 TI - [The use of screens in the anti-tsetse campaign in the forest zone]. AB - After preliminary studies on the ecodistribution of Glossina palpalis s.l. in the sleeping sickness focus of Vavoua (Ivory Coast) trials with screens impregnated with decamethrine were carried out. Artificial supports for insecticide present several advantages: less pollution, less costly, simplicity, facility of use by local personnel, possibility of re-use, integration with other methods. On the other hand, its action based on the attraction of flies which come into contact with the screen is relatively slow, particularly in obtaining a complete interruption of reproduction. The technique is compared with other control methods as the use of biconical traps and selective spraying along borders. Its action can be improved by the addition of attractive host odours or sex pheromones. PMID- 6291200 TI - [Surface morphology of normal and virus-transformed cells in suspended state and their concanavalin A agglutinability]. AB - Suspended cells are heterogeneous in respect to their surface microrelief. The distribution of different microrelieves varies in different cultures. It depends on the mode of cell detachment from the substrate - by EDTA or trypsin. Oncogenic transformation is accompanied by both the increase and decrease of microvillous microrelief. There is no correlation between the surface morphology of transformed cells and their agglutinability by concanavalin A. The treatment with trypsin results in the increase of both agglutinability by concanavalin A and microvillious microrelief. PMID- 6291202 TI - Histological typing of breast tumors. PMID- 6291201 TI - [Turnover of potential-dependent ion channels in the membrane of neuroblastoma cells studied using the protein synthesis inhibitor cycloheximide]. AB - The influence of cycloheximide on currents through sodium and potassium channels in dialysed neuroblastoma cells of clone N18A-1 has been studied under voltage clamp conditions. When cells are incubated with cycloheximide, the conductance of sodium decreases, whereas that of potassium changes only slightly, if at all. The cycloheximide concentration, at which sodium conductance is reduced by 50% during 24 hours incubation, was about 0.5 microgram/ml. The half-time of inhibition at the concentration of 15 microgram/ml was about 9 hours, with the time of 50% inhibition of sodium being more complex than single exponential. The density of sodium channels in control and after the maximal inhibition was estimated to be about 25 and 2.2 micron-2, respectively. The sodium-potassium selectivity and the gating mechanism parameters were shown to be unchanged following cycloheximide treatment. PMID- 6291204 TI - [Teleradiography and tomography in the search for pulmonary metastases in 132 cases of soft tissue and bone neoplasms]. AB - In recent years whole lung tomography (WLT) has been considered mandatory in the staging of some neoplastic diseases, particularly of soft tissues and bone tumors. Since WLT is an exacting roentgen examination, its usefulness was evaluated by analysing 132 consecutive patients submitted to orthogonal chest roentgenograms and WLT, from January 1979 to October 1981 at the National Cancer Institute of Milan. A comparison between WLT and traditional chest X-ray examination was performed in order to evaluate how much tomography is significantly useful in improving diagnosis of lung metastases. From this analysis, the authors conclude that WLT is generally unnecessary when chest roentgenograms are negative, whereas it is useful when single or multiple lung metastases are already evident by traditional X-ray examination. In these cases WLT allows a more accurate identification of the metastases from the point of view of their number and site, which is important not only for the treatment choice but also for the check of its effectiveness. PMID- 6291203 TI - Relationships between morphology and viral etiology in mammary tumors of BALB/cfC3H and BALB/cfRIII mice. AB - The morphologic pattern of mammary tumors induced by milk-transmitted C3H and RIII murine mammary tumor viruses (MuMTVs) in BALB/c hosts infected by foster nursing were investigated further an compared taking into account several tumor and host variables. Comparison of BALB/cfC3H and BALB/cfRIII mammary tumors under identical tumor and host conditions showed highly significant differences in subgross morphology and histologic structure. Differences in cytology were also observed. Data suggest that mammary tumor morphology in high cancer strain mice is controlled by the causative MuMTV. PMID- 6291206 TI - [Separation and direct quantitative estimation of adenine nucleotides on silufol]. AB - A chromatographic method is suggested for separating adenine nucleotides on silufol UV-254 plates in the dioxane-isopropanol-ammonia-water (4:2:1:4) system and for their direct estimation in the thin layer by fluorimetric scanning of chromatograms. The method is applied in estimating the adenine nucleotides content in the liver of intact rats and animals with the fixated stress. PMID- 6291205 TI - [Binding of gamma-aminobutyric acid in the synaptic membranes of rat brain]. AB - The Na+-dependent and Na+-independent (receptor) binding of [3H] GABA is measured in rat brain synaptic membranes. The receptor binding is presented by at least 3 types of binding sites differing in kinetic parameters (dissociation constants and maximal concentration of binding sites). Phenyl-GABA and pyracetam do not affect receptor binding while 4-hydroxybutyric acid, apamine and bicuculline inhibit it with inhibition constants K1=100, 2 and 1 micron, respectively. Solubilization of synaptic membranes with 2% potassium cholate in the presence of total brain phospholipids does not inactivate [3H] GABA receptor binding. PMID- 6291207 TI - [Effect of nicotinic acid on phosphodiesterase and adenyl cyclase activities in chicken liver]. AB - Stimulation of fatty acid biosynthesis by feeding high-carbohydrate diet to chickens after fasting induces a 38% and 23% rise in the 3':5'-cyclo-nucleotide phosphodiesterase and adenylate cyclase activities in the liver tissue, respectively. Administration of nicotinic acid (150 mg per 1 kg of weight) to chickens with stimulated biosynthesis of fatty acids leads to a sharp (70% at an average) decrease of 3':5'-cyclo-nucleotide phosphodiesterase activity, the adenylate cyclase activity under these conditions remains practically unchanged. The participation of the adenylate cyclase system in regulating the fatty acid biosynthesis under mentioned conditions, the role of phosphodiesterase are discussed. PMID- 6291208 TI - [Adenylate kinase activity of the blood of irradiated rats]. AB - The direct and reverse adenylate kinase reactions were studied in blood of rats 1, 3, 7, 15 and 30 days after total X-ray irradiation with a dose of 154.8 mC/kg (600 R) dose. It is found out that adenylate kinase (CE 2.7.4.3) activity in leucocytes and erythrocytes is inhibited on the 3d, 7th and 15th days of the study. Inhibition of the reverse adenylate kinase reaction is pronounced to a greater extent than that of the direct one. The adenylate kinase of erythrocytes is more stable to the effect of ionizing radiations as compared to this enzyme of leucocytes. When the enzymic activity in the blood cells is inhibited, the activity of the direct adenylate kinase reaction in the serum increases significantly and that of the reverse one remains unchanged. PMID- 6291209 TI - [Role of Ca2+ in phenomena of synaptic plasticity]. PMID- 6291211 TI - [Energy metabolism in subcellular fractions of normal and acute ischemic kidneys]. AB - The level of oxidative phosphorylation, activity of phosphofructokinase, fructose 1,6-diphosphate aldolase, ketose-1-phosphate aldolase, glucose-6-phosphatase and lactate dehydrogenase are determined in subcellular fractions in the kidney cortex layer of rabbits which have suffered from acute ischemia (for 15, 30, 60, 120 min). Ischemia inhibits the oxidative processes in mitochondria which is proportional to the duration of the effect. An increase in the activity of glycolytic chain enzymes in microsomes and soluble fraction for 15-30 min of ischemia evidences for a compensation of the energy losses at the expense of glycolysis with short periods of ischemia. Glycolysis is inhibited with a more prolonged effect. It is established that the anti-ischemic protection of the organ viability is to be conducted not only with allo- but also with auto transplantation of the kidney in case of short acute ischemia. PMID- 6291210 TI - [Some physicochemical properties of native and polymerized glutaraldehyde-treated horse heart cytochrome c]. AB - Glutaraldehyde treatment does not change the absorption of cytochrome c either in the visible or in UV spectra. It brings about the formation of dimers, trimers and high-polymeric forms of cytochrome c and shifts the pI of all cytochrome c isoelectric fractions to more acid pH. Polymerization also results in changes of kinetic parameters of cytochrome c benzidine reaction increasing its affinity to 3,3-diaminobenzidine with a simultaneous decrease in the effectiveness of H2O2 binding. These biochemical changes can be related to immunochemical differences of native and glutaraldehyde-treated cytochrome c. PMID- 6291212 TI - [Metabolic aspects of temperature adaptation by fish]. AB - During 8-hour acclimation to changes in the water temperature by 10 degrees within the range of 20-30 degrees C the metabolic compensation of the temperature effects in the carp liver mitochondria is manifested at the level of thermogenesis, activity of succinate dehydrogenase and rate of oxidative phosphorylation. No temperature compensation is found for the activity of cytochromoxidase, ATPase, rate of nonphosphorylating oxidation, content of ATP, phosphorus and calcium in the mentioned organelles. PMID- 6291213 TI - [Glomus jugulare tumor]. PMID- 6291214 TI - [Cystosarcoma phyllodes of the breast]. PMID- 6291215 TI - Solid-state radiation chemistry of DNA and its constituents. AB - The molecular structure as well as the mechanisms of formation and decay of free radicals produced in DNA and its constituents by ionizing radiation is reviewed. Starting with the description of the spectral parameters for cations and anions in natural nucleic acid bases, emphasis is given to the comparable species formed in the group of the 5-halogen substituted uracil derivatives. The consequences of the attachment of a ribose or ribosephosphate group to the bases is discussed in terms of the distribution of primary radicals which, again, is shown to be different from those of the subunits in DNA itself. The quantitative aspects of radical formation are discussed in terms of G values and their dependence on the temperature of irradiation. Finally, a schematic presentation of the major modes of radical reactions is given occurring upon warming of the primary species in the DNA subunits and in DNA itself. PMID- 6291216 TI - Collagenase for Peyronie's disease experimental studies. AB - This pilot study was designed to test the feasibility of using purified clostridial collagenase in the clinical management of Peyronie's disease. The basic properties of this agent are discussed. We studied its effect on Peyronie's plaque tissue by a quantitative in vitro assay utilising the liberation of free alpha-amino groups as an index of enzymatic collagenolysis. Tissue from three patients with Peyronie's disease was used. Tunica albuginea from a second group of three normal patients was studied in the same manner, and no selectivity for the collagen of Peyronie's plaques was identified. Utilising human pericardium as a uniform collagenous substrate, a simple dose-effect relationship was established, and the distribution characteristics of injected collagenase observed. Its effects on blood vessels and nerves in vivo was determined as well as the effects of collagenase on the histology of normal and diseased human tissue in vitro. A tentative dose for use in Peyronie's disease was established, which is discussed in light of existing toxicological data. The study was designed to test the feasibility of purified collagenase in the clinical management of Peyronie's disease. Data included detail plaque digestion and dose effect relationships in vitro, as well as the histological effects on plaques, blood vessels, and nerves in vivo and in vitro. It is concluded that collagenase may warrant further clinical testing in the treatment of Peyronie's disease. PMID- 6291217 TI - [Detection of antibodies against the virus causing viral gastroenteritis in swine using the modified autoradiography test]. AB - A modified method for autoradiographic determination of antibodies to the virus of transmissible gastroenteritis of swine was developed. It is based on the actual reaction between antigen bound in cells of the infected cell cultures and antibodies in tested sera, which is visualized, by help of rabbit antibodies marked 125J (125J RaSw IgG antibody) to porcine IgG, on a sensitive radiograph and evaluated on the basis of darkening at the point of positive immunological reaction. Specificity of the test and mutual comparability and reproducibility of the results were confirmed by examining the known positive and negative sera and by comparison with the results of the virus-neutralisation test. Out of the 36 examined porcine blood sera the antibodies were proved autoradiographically only in the samples positive also by virus-neutralisation. In experimentally infected pigs, the same dynamics of antibody production was recorded by the two tests. They were, however, demonstrated autoradiographically already by the two tests. They were, however, demonstrated autoradiographically already the eighth day after infection, while by virus neutralisation test as late as 14th day. Their level increased gradually until 35th day after infection. PMID- 6291218 TI - [Detection of leukosis on brown Leghorn chicken farms]. AB - Samples taken on the Brown Leghorn hen farms were tested for the leucosis virus. Out of 21 samples the leucosis virus was present in five cases (24%). Viruses are detectable in chicken embryonic cells, accompanied by transformation and focus formation. Viruses are not transformed by the guniea pig and quail embryonic cells. The results were achieved by fusion but not by co-cultivation of the tested cells with permanent line 16 Q. These viruses appear to belong to the sub group B, it is less probable that they belong to the sub-group C. The isolated viruses document spontaneous infection of the originally leucosis-free hen farm during five years when leucosis-free conditions were not secured. PMID- 6291219 TI - Suspected ammonium nitrate fertiliser poisoning in cattle. PMID- 6291220 TI - Oestrone sulphate in goats' milk. PMID- 6291221 TI - Serological analysis of recent type O foot-and-mouth disease virus isolates from Europe. PMID- 6291222 TI - [Case of alveolar cell adenocarcinoma of the lungs]. PMID- 6291223 TI - [Extraction and freeze-drying of herpes virus C3-1 isolated from turkeys]. AB - Possibilities for producing dried preparations of turkey herpes virus C3-1 by use of various ultra-sound desintegrator-systems and different ways of processing the cell suspensions containing the virus were studied. It was established that cell less virus of a 1.5 to 2 log smaller quantity than the initial virus containing cell suspension can be produced by ultrasound generator with an indirect action - a vibrating membrane, which transmits the impulses through a liquid medium and the container's walls on the object. The use of a generator with direct action -- titanic probe placed into the suspension results in an insignificant loss and in some cases even leads to higher titer. In the process of freeze drying the protective media SPGA, 5% pepton and 5% horse serum produce almost equal results. Highest dried virus yield was produced by lyophilization of entire (without centrifugal separation) ultrasounded cell suspension. In such cases the titer of the freeze-drying virus was by 0.2-0.4 log higher than the initial cell virus determined after the microplate method. The authors are of the opinion that titration by this method does not give an exact criterion for comparison of virus content in frozen cell and freeze-dried preparations. A proper protection against Marek's disease was established in laboratory and terrain experiments with chickens following immunization by freze-dried preparations of the strain C3-1 (losses were reduced 3-4 times). An immunization dose of 2000 POU lyophilized virus is recommended for practical use. PMID- 6291224 TI - [Control of enzootic Aujeszky's disease on pig farms]. AB - Virologic and serologic investigations were carried out to demonstrate the persistence of the virulent virus of Aujeszky's disease. Threefold vaccinations were conducted with a live vaccine, embracing all animals on the farm and those that were born during the time when sanitation measures were applied. The first vaccination of the sucking pigs was performed with 10 doses of the vaccine in each case. During the time of applying sanation measures the farm was kept closed, the pigs after weaning being transferred to another farm. After sanitation was over some new virologic and serologic studies were carried out along with the serologic examination of nonimmune pigs that had stayed on the farm. These investigations did not prove the presence of a wild virus on the farm. It was found that the manifold vaccinations and use of higher amounts of the vaccine might well suit the base in working out sanitation programmes for farms infected with Aujeszky's disease. PMID- 6291225 TI - [Analysis of the immunologic properties of foot-and-mouth disease vaccines]. PMID- 6291226 TI - [Control of latent enzootic Aujeszky's disease]. AB - An attempt was made on a swine-breeding complex, where the virus of Aujeski's disease was demonstrated to interrupt the inapparent epizootic process of the disease. The persistence of the virus on the complex was demonstrated via virologic investigations of organs of pigs with respiratory symptoms and studies of sera of various groups of animals. A batch of pigs was divided into three groups of which one was left untreated, one was vaccinated at the time of weaning, and one was vaccinated at the age of 14 days, using as many as 10 vaccinal doses with one dose of a live vaccine being used at the moment of weaning. During fattening the pigs were kept in isolation and under clinical observation with the study of organs and sera. Wild virus was demonstrated only in the group of twice vaccinated pigs. It was established that the manifold vaccinations of sucking pigs on farms that were enzootic in terms of Aujeszky's disease, using a live vaccines, with the isolation of the animals right after weaning could contribute to the interruption of the inapparent epizootic process. PMID- 6291227 TI - [Effect of colostral antibodies on the development of immunity in piglets immunized with live vaccine]. AB - Pigs of sows treated with a live vaccine were in turn vaccinated with a live vaccine at different age. Part of them were challenged with a virus at 60 days of age, the remaining -- at 90 days. The pigs were kept under clinical observation, and were killed on the 10th day following challenge in order to be thoroughly investigated. It was found that the passive immunity (virus-neutralizing antibodies) in pigs of twice (live vaccine) treated sows persisted for 4 to 5 weeks after birth. It dropped steplike, however, the interference of the vaccinal virus speeded up the process -- the earlier the vaccination the faster the exhaustion of immunity. The passive immunity was found to neutralize the vaccinal virus and hamper the building of active immunity. The presence of eosinophile leukocytes in the lymph nodes, tonsils, and spleen of the vaccinated pigs spoke of an immune reaction. PMID- 6291228 TI - The differential diagnosis of testicular germ cell tumors in theory and practice. A critical analysis of two major systems of classifiction and review of 389 cases. PMID- 6291229 TI - Transplantation of ACTH-secreting pituitary tumor cells in athymic nude mice. AB - Chronic excess of glucocorticoids results in Cushing's syndrome in humans. A common cause of excess cortisol secretion is the presence of an adrenocorticotropin secreting pituitary tumor which stimulates the adrenal cortex to produce excess glucocorticoids. ACTH-secreting AtT-20 mouse pituitary cells transplanted subcutaneously in oestrogenized athymic nude mice form tumors rapidly. Six weeks after receiving the tumor transplants, the mice weighed 45% more than normal mice due to the increase in body fat. The tumor-bearing mice exhibit the familial "buffalo hump" appeaance due to the abnormal distribution of body fat. The adrenal glands of the tumor-bearing animals are enlarged due to hypertrophy of the zona fasciculata. The foamy looking fasciculata cells in normal mice were converted to dense, eosinophilic cells in the tumor-bearing mice. Transplantation of normal pituitary glands to athymic nude mice with or without oestrogen treatment did not produce these morphological changes. The experimental model described here may be useful for future studies of Cushing's syndrome. PMID- 6291230 TI - Benign pleomorpic adenoma arising in a parotid lymph node. PMID- 6291231 TI - Isolation and characterization of bacteriophage T3 mutants sensitive to restriction by EcoRI. PMID- 6291232 TI - Mutants of encephalomyocarditis virus requiring a hypertonic environment for optimal growth in HeLa cells. PMID- 6291233 TI - Inhibition of Epstein-Barr virus-associated nuclear antigen (EBNA) induction by (2',5')oligoadenylate and the cordycepin analog: mechanism of action for inhibition of EBV-induced transformation. PMID- 6291234 TI - Low interferon binding activity of two human cell lines which respond poorly to the antiviral and antiproliferative activity of interferon. PMID- 6291235 TI - A pH-dependent antigenic conversion of empty capsids of poliovirus studied with the aid of monoclonal antibodies to N and H antigen. PMID- 6291236 TI - Characterization of a temperate bacteriophage for Azospirillum. PMID- 6291237 TI - Transformation parameters of chick embryo fibroblasts transformed by Fujinami, PRCII, PRCII-p, and Y73 avian sarcoma viruses. PMID- 6291238 TI - Heterogeneity in the structural glycoprotein (VP7) of simian rotavirus SA11. PMID- 6291239 TI - Excision of polyoma virus DNA from that of a transformed mouse cell: identification of a hybrid molecule with direct and inverted repeat sequences at the viral-cellular joints. PMID- 6291241 TI - [Clinical picture and diagnosis of malignant non-epithelial tumors of the stomach]. PMID- 6291240 TI - Thyroid-stimulating autoantibodies. PMID- 6291242 TI - [Anaerobic infection]. PMID- 6291243 TI - [Mechanism of action of laser radiation on the neuronal somatic membrane]. PMID- 6291244 TI - [Research to improve smallpox vaccine: the contribution to the elimination of smallpox in the world]. PMID- 6291245 TI - [Transposon nature of retroviridae and carcinogenesis]. PMID- 6291246 TI - [Morphological, cytogenetic and proliferative characteristics of Syrian hamster HTC-2 and HTC-1 cells and of the HTCT tumor cell line transformed by herpes simplex type 2 virus]. AB - Morphological, caryological, cytoproliferative, and isoenzyme studies of Syrian hamster cells HTC-2 and HTC-1 transformed by herpes simplex virus type 2 and a HTCT line of tumor cells were carried out. The hamster origin of these cell lines was established by chromosomal analysis and electrophoretic mobility of lactate dehydrogenase and glucose-6-phosphate-dehydrogenase isoenzymes. The transformed and tumor cell lines differ from normal cells by altered morphology, an aneuploid chromosome set and increased proliferative activity. Electron microscopic examination of transformed cells revealed increased amounts of filamentous and tubular structures in the cytoplasm. No retroviruses were found. PMID- 6291247 TI - [Encephalomyocarditis in hamadryas baboons]. AB - In 1978--1979, four cases of encephalomyocarditis were recorded in baboons in a simian preserve of the USSR AMS Research Institute of Experimental Pathology and Therapy located in the Zapadnyaya Gumista river valley. The diagnosis was made on the basis of virological and pathomorphological examinations. The isolate recovered from the dead monkeys was identical with the prototype strain of encephalomyocarditis virus (EMC) and the EMC-70 strain previously isolated by us from M. rhesus monkeys. The infection of monkeys caused by the EMC-78/79 virus involved a severe affection of the myocardium of the type of parenchymatous interstitial myocarditis, moderate encephalitis in the central nervous system, inflammatory changes in the brown fat and cross-striated muscles. An experimental infection in guinea pigs and mice exhibited similar features. PMID- 6291248 TI - [Spontaneous cytomegalovirus infection of kidney cell cultures of green monkeys]. AB - In subcultivation of 104 batches of primary kidney cell cultures from clinically normal green monkeys 66 batches (47%) showed cytopathic changes indicating spontaneous virus contamination of the cultures. Sixty-six strains were isolated and the properties of 23 cytopathic strains were studied. Electron microscopic examinations of 6 virus isolates identified them as members of the herpesvirus group. Serological examinations of 335 clinically normal green monkeys demonstrated antibodies to one of our virus isolates (No. 8532) in 192 (57.3%) animals. Virus strain 8532 was neutralized by antiserum to green monkey cytomegalovirus M-1. The isolates obtained in the present study exhibit the properties previously unknown for green monkey cytomegalovirus such as the capacity for replication in rabbit, sheep, cow, and piglet cell cultures. PMID- 6291249 TI - [Cytomegalovirus isolation in chronic salivary gland diseases of adults]. AB - The results of virological investigation of a rare clinical instance of familial disease with a localized form of cytomegalovirus infection which, as a rule, is not diagnosed in life are presented. The infection localized in the salivary glands was the source of generalization of the process with a high fever, eruptions, lymphadenopathy, inflammatory changes in the excretory ducts of the salivary glands. Exacerbation of the chronic process was accompanied with high antibody titres (1:256--1:1024) with inhibition of the cellular immunity reactions in the mother. The latent form of infection in the father and two daughters ran without clinical manifestations, with low antibody titres and moderate inhibition of cellular reactions. The isolated strains of cytomegalovirus were identical in all the subjects under study and antigenically close to the Ad-169 strain. PMID- 6291251 TI - [Experimental study of ophthalmic embryopathy of herpetic etiology]. AB - The teratogenic effect of herpes simplex virus was studied in white rats and in organ cultures of rat embryo eye rudiments. In herpetic infection, changes in the lens fibers, delayed formation of the vitreous body cavity, disturbed differentiation of the retina, appearance of typical and atypical colobomas were observed significantly more frequently than in controls. The models used in the study are suitable for experimental investigations of the pathogenesis of ophthalmoembryopathies of herpetic etiology. PMID- 6291250 TI - [Modeling of mixed infection by tick-borne encephalitis and Powassan viruses in mice]. AB - Simultaneous inoculation of mice with tick-borne and Powassan viruses was shown, depending on experimental conditions, to result either in stimulation of infection or its unchanged course as compared with monoinfection and inoculation with the viruses at 2--3-week intervals in cross protection of mice against the superinfecting virus. Simultaneous inoculation of mice with the two viruses was accompanied by their multiplication in the blood and brains of mice and formation of antihemagglutinating antibodies to each of them. In the virus population in the brains of mice there was either formation of a mixture of two viruses or their phenotypic mixing. In cross protection, multiplication of the superinfecting virus in the blood and brain of mice was slightly inhibited, the antihemagglutinating antibody to a second virus either did not form or appeared in low titres. PMID- 6291252 TI - [Simple method of isolating herpes virus DNA]. PMID- 6291253 TI - [Comparative evaluation of the CFT and passive hemagglutination reaction in diagnosing ophthalmic herpes]. PMID- 6291254 TI - Occupational asthma. PMID- 6291256 TI - The value of artificial beta cell in the management of insulinoma. PMID- 6291255 TI - Calcium channel blockers. AB - Calcium (Ca(++)) plays an essential role in many cardiovascular physiologic processes. Electrophysiologic properties of the sinus and atrioventricular nodes greatly depend on Ca(++) ion influx. Also, Ca(++) is the main link for excitation contraction coupling of the myocardium. Ca(++) channel blockers are a group of heterogeneous compounds that block the ionic influx of Ca(++) into the myocardial and vascular smooth muscle cells. Because Ca(++) plays a central role, it is not surprising that Ca(++) channel blockers can produce profound alterations in cardiovascular functions. Recently several studies have shown these agents to be useful in the treatment of supraventricular tachyarrhythmia, variant angina, chronic stable angina and hypertrophic cardiomyopathy. In the future they may be found useful in preserving myocardium during cardiopulmonary bypass, in limiting infarct size and in the treatment of hypertension and congestive heart failure. PMID- 6291257 TI - Association of myoglobinuria with adenovirus infection. PMID- 6291258 TI - [Case of delirium tremens and acute atypical alcoholic neuropathy]. PMID- 6291259 TI - [Progress in the chemotherapy of ovarian cancer]. PMID- 6291260 TI - Metabolism and excretion of benzo[a]pyrene in the rabbit. AB - 1. Following i.v. administration of [14C]benzo[a]pyrene (3 mumol/kg) to rabbits, 30% of the 14C dose appeared in bile and 12% in urine, within six hours. 2. Biliary and urinary metabolites were mainly conjugated; less than 12% of the 14C was extractable with ethyl acetate, but after treatment with beta-glucuronidase or aryl sulphatase 30-40% became extractable. 3. H.p.l.c. analysis of the extracts indicated that the major non-polar metabolite was benzo[a]pyrene, 9,10 diol (18% of 14C in bile and 24% of 14C in urine, mainly conjugated with glucuronic acid). Smaller amounts of the 4,5-diol, the 3,6-quinone, and the 9 hydroxy- and 3-hydroxybenzo[a]pyrene were also found in bile (total less than 10%), together with 9-hydroxybenzo[a]pyrene and two unknown metabolites (X and Y) in urine (total less than 4%). 4. The proximate carcinogen, the 7,8-diol, was not detected in any extract. 5. After intraduodenal administration of biliary metabolites of [14C]benzo[a]pyrene (approx. 0 X 3 mumol), 14C was excreted in the bile (21% dose) and urine (14%) within 23 h, indicating that metabolites can undergo enterohepatic circulation in the rabbit. PMID- 6291261 TI - Conduction studies of the saphenous nerve in normal subjects and patients with femoral neuropathy. PMID- 6291263 TI - Interaction of bacteriophage T7 with Hind-endonuclease-producing Haemophilus influenzae Rd cells. AB - In order to test the influence of the restriction endonucleases of Haemophilus species on in vivo DNA restriction of phage T7 and its close relative T3 as well as to determine whether the anti-DNA restriction effect of the T7 and T3 ocr+ gene is also expressed in Haemophilus cells, we studied the interaction of T7 and T3 with Haemophilus influenzae serotypes Rd and Rc and Haemophilus parainfluenzae. The results show that T7 and T3 are unable to infect these bacterial species because they do not absorb to the cells. PMID- 6291262 TI - Erythropoietin-mediated erythrocytosis in rodents after intrarenal injection of nickel subsulfide. AB - Rats and guinea pigs developed pronounced erythrocytosis at one to four months after unilateral intrarenal (ir) injection of nickel subsulfide (Ni3S2). For example, at two months after ir administration of Ni3S2 (5 mg) to rats, blood hematocrit values averaged 70 +/- 3 percent (p less than 0.001 vs. 48 4/- 2 in controls); at two months after ir administration of Ni3S2 (20 mg) to guniea pigs, blood hematocrit values averaged 67 +/- 6 percent (p less than 0.001 vs. 49 +/- 1 percent in controls). Hamsters and gerbils did not develop erythrocytosis after ir injection of Ni3S2 (5 mg/animal). Administration of Ni3S2 to rats by intrasplenic injection did not increase blood hematocrit; splenectomy did not prevent erythrocytosis in rats that received ir injection of Ni3S2. Erythrocytosis in rats was completely blocked by excision of the Ni3S2-injected kidney but was unaffected by excision of the non-injected kidney. Partial inhibition of Ni3S2-induced erythrocytosis in rats occurred after simultaneous ir injection of Mn, Cu, or Al dusts, benzo(a)pyrene, or subcutaneous (sc) infusion of sodium diethyldithiocarbamate. Erythrocytosis induced by ir injection of Ni3S2 was augmented by ir injection of Cr dust or intramuscular (im) administration of iron-dextran. Erythrocytosis occurred in rats after ir implantation of Ni3S2 within semi-permeable cellulose tubules, indicating that phagocytosis of Ni3S2 particles is unnecessary for erythropoietic stimulation. Erythropoietin (Ep) activity in rat serum increased sixfold at two weeks after ir injection of Ni3S2 (p less than 0.001 vs. controls), but Ep activity in pooled extracts of Ni3S2 treated rat kidneys did not increase significantly. This study identifies several factors that influence erythropoietic stimulation by Ni3S2, and furnishes salient information concerning the pathogenesis of Ni3S2-induced erythrocytosis. PMID- 6291264 TI - [Lung changes induced by copper sulfate. An experimental contribution to the so called "vineyard sprayer's lung"]. AB - Vineyard sprayer's lung is an occupational lung disease, caused by inhalation of Bordeaux mixture (1-2% aqueous solution of copper sulfate, neutralized with hydrated lime). Experimental studies in mice after a longstanding exposition to copper sulfate aerosol showed focal accumulations of macrophages in alveoli with different cytoplasmatic inclusions, focal interstitial granuloma formation, increased formation of collagenous and elastic fibers, and increased production of surfactant phospholipids, phagocytosed by macrophages. As a conclusion of these results, copper seems to be the responsible agent for the development of lung changes, seen in vineyard sprayer's lung. PMID- 6291265 TI - [Lung adenomatosis--adenocarcinoma]. PMID- 6291266 TI - The effects of leucine-enkephalin on the spontaneous motility of the circular muscle of the cat colon. AB - Since the effects of endogenously occurring opiates in the regulation of colonic motility are unknown, we set out to study the actions of leucine-enkephalin on the spontaneous contractile and electrical activity of the isolated circular muscle of the cat colon. The muscle strips exhibited regular rhythmic contractions and myoelectrical slow waves 3.67 +/- 0.16 (SEM) times/min. Mean contraction force was 1.20 +/- 0.09 mN. Leucine-enkephalin caused a dose dependent increase in contractile activity by a factor of 8.24 +/- 2.68; ED50 was 2.23 X 10(-9) M, ED100 4.33 X 10(-8) M. Methionine-enkephalin was equally effective, but morphine was much less effective. Naloxone acted as a specific antagonist. Concentrations of 10(-8) and 10(-7) M shifted the dose-response curve to the right, at the ED50 of leucine-enkephalin by 0.66 and 1.63 log units, respectively. Naloxone exhibited the characteristics of competitive antagonism; pA2 was -8.69, the slope was 1.06. Other antagonists did not affect the enkephalin actions on the muscle. The results of this study suggest that leucine enkephalin acts on specific receptors of the circular muscle of the cat colon. It is conceivable, therefore, that leucine-enkephalin exerts its effects under physiological conditions to increase segmenting motor activity. PMID- 6291268 TI - Regulation of the sarcoplasmic reticular Ca2+ transport ATPase by phosphorylation and dephosphorylation. AB - At 0.1 mg/ml protein and 0.45 micrometers free Ca2+ l mol trichloroacetic acid precipitable phosphate is incorporated into 100,000 g SR protein as hydroxylamine sensitive acylphosphate. At nearly physiological protein concentration (ca. 7 mg/ml) a total of ca. 0.8 mol phosphate/100,000 g protein is incorporated, from which a fraction of 0.3 mol/100,000 g protein is insensitive to the hydroxylamine treatment, i.e. it is alkylphosphate. Phosphorylase kinase accelerates the alkylphosphate incorporation ca. 3-fold and enhances its final level to 0.7 mol/100,000 g protein. At 1.6 nm free Ca2+ alkylphosphate incorporation occurs at high SR concentration to a maximal extent of 0.5 mol/ 100,000 g protein. The incorporated alkylphosphate is present in comparable amounts in the 100,000 Mr Ca2+-transport ATP-ase and a polypeptide of Mr 9,000. PMID- 6291267 TI - [Ketoconazole. 3 years' experience with a new orally active broadspectrum antimycotic]. AB - Ketoconazole is an orally high effective broad-spectrum antimycotic of the imidazole series. It is very well tolerated and stands out by its long-lasting therapeutic serum levels, lack of development of resistant fungi, simple administration and low toxicity. Own experience has shown its high efficacy in pityriasis versicolor, mycoses due to dermatophytes, skin and mucous membrane candidoses and chronic mucocutaneous candidosis syndromes. Its effect in systemic mycoses is promising. Consistent follow-up of laboratory data did not suggest any changes due to ketoconazole even after more than one year of treatment. PMID- 6291269 TI - Comparative studies on the ATPase-binding sites in Ca2+-ATPase and (Na+ + K+) ATPase by the use of ATP-analogues. AB - The effects of ATP-analogues on Ca2+-ATPase and (Na++ K+)-ATPase have been studied. The participation of sulfhydryl groups in the recognition of ATP by both transport ATPases is indicated by the fact, that the disulfide of thioinosine triphosphate inactivates both enzymes. The reactivity of rapidly and slowly reacting sulfhydryl groups in the ATP binding sites of both enzymes is altered by the presence of transport substrates. At lease in (Na+ + K+)-ATPase Na+ and Mg2+ appear to alter the structure of the ATP binding site, which conclusion is fortified by the fact, that the photoinactivation of the enzyme by 3'-O-[3-(2 nitro-4-azidophenyl)-propionyl]-ATP needs Mg+. Chromium(III)ATP, a MgATP analogue, inactivated both transport ATPases by the formation of a stable chromo phosphointermediate. In the case of Ca2+-ATPase this was concomited by the occlusion of Ca2+ in a stable form. No occlusion of Na+ was observable so far in the (Na+ + K+)-ATPase. Contrary to the expectation of the Albers-Post-scheme the hydrolysis of the phosphointermediate formed from chromium(III)ATP was protected by K+, but activated by high concentrations of Na+. Consequently, despite of the inhibition of (Na+ + K+)-ATPase activity, chromium(III)-ATP supported the Na+ - Na+-exchange reaction in everted red blood cells. PMID- 6291270 TI - Electromechanical coupling II. The effect of perchlorate upon excitation contraction coupling in frog skeletal muscle fibres. AB - In single skeletal muscle fibres perchlorate causes a large shift of the potential dependence of contraction activation to more negative potentials without a corresponding alteration in the kinetics of the inactivation process. PMID- 6291271 TI - [5-Methylcytosine content in erythroleukemic cells of rats after induction with dimethylsulfoxide]. AB - Erythroleucemic cells of rats were induced by DMSO for the terminal erythroid differentiation. The base composition and the 5 MC content of the DNA were analyzed by HPLC and a significant difference between non-induced and induced cells was not found. The analysis of labelled DNA by the restriction enzymes Hha I, Hpa II, and Msp I did not result in a different pattern before and after the induction. These results suggest that the 5 MC content of the DNA does not change during the induced terminal erythroid differentiation. PMID- 6291272 TI - Subcellular distribution and stability of the major hemolytic activity of Entamoeba histolytica trophozoites. AB - Since the hemolytic activity of extracts from Entamoeba histolytica trophozoites previously described by us might determine at least partially the necrotic lesions of amebiasis, we have continued its characterization in vitro. Using rat erythrocytes as target cells, we have found that cytolysis by E. histolytica trophozoite extracts was (1) dose dependent, (2) localized mainly in a vesicular fraction whose absolute and specific activities were respectively 1.9 and 4.0 times higher than those of total extracts, (3) maximal at pH 8 in the presence of 1 mM Ca++, and (4) progressively lost by heating at 90 degrees C or repeated freezing and thawing. From these results we infer that the major hemolytic factor of E. histolytica may be a protein normally neutralized by an intracellular inhibitor or activated during fractionation. PMID- 6291273 TI - On culture of Entamoeba histolytica in plastic tissue culture flasks. PMID- 6291274 TI - [Oncological after care. The involvement of the general practitioner as education object of general medicine]. PMID- 6291275 TI - [Role of cholinergic transmission of inhibitory and disinhibitory effects in mechanisms of reinforcement and internal inhibition]. PMID- 6291276 TI - [Effect of alcohol on the system of reinforcement]. PMID- 6291277 TI - [Physiological interpretation of statistical signs of the relation between the spike-trains of interacting neurons]. PMID- 6291278 TI - [Neuronal mechanism of temporary connections]. PMID- 6291279 TI - [Stability of interneuronal connections in the sensomotor cortex of the cat]. PMID- 6291280 TI - [Background activity of cortical neurons before and after a burst of impulses]. PMID- 6291281 TI - [Excitability of the electro-excitable membrane of a cortical neuron after its intracellular tetanization by a hyperpolarizing current]. PMID- 6291282 TI - [Soluble and insoluble suture materials for non-mucous membranes. Long-term animal experiment study]. PMID- 6291283 TI - Transmission of mouse-pox in colonies of mice. PMID- 6291284 TI - [Organization of rehabilitative therapy of patients with nervous system diseases in specialized departments of urban polyclinics]. PMID- 6291285 TI - [Hereditary predisposition as a risk factor for the development of ischemic stroke]. PMID- 6291286 TI - [Differential diagnosis of dysarthria]. PMID- 6291287 TI - Cloning of the mouse satellite DNA. AB - A stable recombinant clone was constructed by inserting a 1.5 kb mouse satellite DNA HindIII restriction fragment into the plasmid pBR-322. The cloned fragment according to both hybridisation data and restriction analysis seems to be identical with the major component of the mouse satellite DNA. It contains two Atu4001 (EcoRII) monomers, one dimer and one "1.5-mer". HindIII restriction sites are either in position around 95 or 140 of the Atu4001 monomer. Our results and the recently published prototype sequence of the mouse satellite DNA Sau961 monomer (15) suggest that HindIII cleavage of the mouse satellite DNA follows the B type restriction pattern. PMID- 6291288 TI - Binding of fluorescent and radiolabelled alprenolol to intact cultured brain cells and liposomes. AB - Beta-adrenergic antagonist L-alprenolol was labelled for radioactivity and UV fluorescence, and the binding to intact cultured chicken brain cells under equilibrium conditions, as the model of the experiments in vivo, was probed. The application of fluorescent label did not explore any cell type or cell particle with enhanced binding. The analysis of the radiolabelled alprenolol-binding revealed a non-specific accumulation on the surface of these cells, which was inhibited by unlabelled alprenolol excess, however, this is the peculiarity of the specific binding. Our results emphasize that for the characterization of membrane receptors of intact cells, the accurate separation of specific and non specific binding is required. The application of a ligand concentration as low as possible and a very effective washing procedure for the removal of the unbound (and the bulk of the nonspecifically bound) label is recommended. PMID- 6291290 TI - Preparation of spin-labeled poly(U) and its activity in assays with eukaryotic ribosomes. AB - ESR studies on the interaction of spin-labeled polynucleotides with ribosomes require a sufficient label-to-nucleotide ratio. Using three different spin labels (SL) we have elaborated a technique to label poly(U) up to a ratio of 1 SL per 30 uridine residues. This ratio is much higher than maximal values obtained by other authors. The SL-poly(U) was shown to have the same activity as unlabeled poly(U) to direct synthesis of poly(Phe). SL-poly(U) binds to rat liver ribosomes in the presence of Mg2+ as shown by ESR. Titration with EDTA leads to a release of SL poly(U) from ribosomes. PMID- 6291289 TI - Mobility and clusterlike organization of liposomal cytochrome P-450 LM2: saturation transfer EPR studies. AB - Rotational diffusion of the electrophoretically homogenenous isozyme cytochrome P 450 LM2 from rabbit liver microsomes has been studied in buffer solution and in phospholipid vesicles by means of saturation transfer EPR spectroscopy. Sulfhydryl groups of the enzyme were selectively modified using a maleimide spin label. The effective rotational correlation time of 220 ns for the rotation of cytochrome P-450 in buffer solution is consistent with the fact that the purified free enzyme occurs as an oligomeric (6-8 monomers) aggregate. Further, the clusters rotate almost isotropically and therefore are in a first approximation spherically shaped. The effective correlation time of about 180 microseconds observed strong immobilization thus evidencing protein aggregation within the membrane. The anisotropic character of the spectra indicates a nonspherical shape and/or anisotropic rotational motion of the cluster. The results are compared with corresponding data from cytochrome P-450 in microsomal form. PMID- 6291291 TI - Relation between Na+-K+-ATPase activity and dopamine release from rat striatum slices. AB - Ouabain inhibits potassium induced dopamine release in a competitive manner. On the other hand, amphetamine induced release is facilitated. In dependence on the stimulus power used, the electrically evoked release is reduced by ouabain. Thus, cardiac glycosides represent an appropriate tool for differentiating pools and processes involved in transmitter release, controlled by Na+-K+ ATPase activity. PMID- 6291292 TI - Separation of the different classes of conjugates formed by metabolism of benzol[a]pyrene in the northern pike (Esox lucius). PMID- 6291293 TI - The significance of quantitative biological cell parameters in the handling of mammary carcinoma. AB - Recent work has shown that in breast cancer patients DNA measurements and estrogen receptor (ER)- analysis can contribute valuable information in the judgement of prognosis and choice of therapy. We report a case of advanced breast carcinoma with extensive liver metastases which clinically was regarded as beyond all therapeutic possibilities. Liver metastases were diagnosed first. Some time later a breast carcinoma was detected which was suggested to be the primary malignancy. Since high ER-values were demonstrated in the breast carcinoma, anti estrogen therapy was given on trial. After three months the tumor mass in the liver had drastically diminished and today, more than 4 years after diagnosis, the patient is free from symptoms and doing excellently. A retrospective, microspectrophotometric DNA analysis in both the breast carcinoma and the liver metastasis showed DNA distribution patterns with nearly all the carcinoma cells exhibiting DNA values in the normal diploid region. This type of DNA profile has previously been shown to be correlated with good prognosis. We conclude that quantitative parameters such as DNA- and ER-content can give prognostic and therapeutic information over the above that furnished by clinical stage and morphological criteria. PMID- 6291294 TI - A clinical view of pain physiology. PMID- 6291295 TI - Effect of brain monoamines on the secretion of adrenocorticotrophic hormone. AB - The role of brain monoamines (5-HT, NA and DA) in the secretion of adrenocorticotrophic hormone (ACTH) was studied in view of contradictory reports. Plasma corticosterone levels and the rate of synthesis of corticosterone in vitro by the adrenal gland were estimated in albino rats and have been taken as the index of ACTH activity. These estimations were done in unstressed and stressed, and in untreated and treated rats. Drugs were administered intracerebroventricularly to the rats to cause selective degeneration of tryptaminergic, noradrenergic or dopaminergic neurons. The results show that plasma corticosterone levels and the rate of synthesis of corticosterone were significantly decreased after selective degeneration of tryptaminergic neurons in unstressed rats. After selective degeneration of either tryptaminergic or noradrenergic neurons, the acute increase in the plasma corticosterone levels and rate of synthesis of corticosterone in vitro by adrenal glands in stressed rats were significantly inhibited. These results have been interpreted to suggest that the central tonic control on adrenal glands may be 5-HT mediated and that during stress ACTH secretion may be both 5-HT and NA mediated. DA does not seem to have significant role in the regulation of ACTH secretion. PMID- 6291296 TI - Thyrotrophin-binding-inhibition assay: comparison of crude and purified membrane preparations. AB - Two different thyroid membrane preparations (TMP), crude and pure, made from a homogenate of normal human thyroid tissue, were used to test purified serum IgG from 36 patients with Graves' disease, and 10 normal control subjects, in the thyrotrophin-binding-inhibition (TBI) assay. All reagents for the assay were identical, and aliquots of each IgG were tested in both TMP simultaneously, under exactly comparable conditions. Blood was drawn while the patients were hyperthyroid (19), euthyroid (13), or hypothyroid (4); 15 of the patients were untreated and 21 were being treated. The frequency of positive TBI was similar in both TMPs and highest among the 131I-treated patients. Comparison of TBI results in both TMP for each IgG sample revealed wide differences, and 47.1% of the Graves' IgGs were TBI positive in one membrane preparation, while negative in the other. There was no correlation of TBI values between the two TMP, or with clinical status, or the presence of standard thyroid antibodies. The findings indicate that the IgGs of Graves' disease, as now tested in the TBI assay, bind heterogeneous to different fractions of the thyroid membranes. PMID- 6291297 TI - Effect of intravenous calcium on urinary cyclic adenosine-3',5'-monophosphate and hydroxyproline in children with hypoparathyroidism. PMID- 6291298 TI - Monitoring ovarian function by a solid-phase chemiluminescence immunoassay. PMID- 6291300 TI - Testicular receptors of human chorionic gonadotrophin in adult men. Binding and degradation of the hormone. AB - Binding and degradation of human chorionic gonadotrophin (hCG) to testicular tissue obtained by biopsy from 9 men with gonadal disorders were investigated. Vacant hCG receptors were assayed in partially purified testicular homogenates using [125I]hCG (radioiodinated with chloramine T). Degradation of [125I]hCG during exposure to human testicular preparations was measured in terms of the ability of supernatants to specifically bind to rat testicular receptors. Binding of [125]hCG was time and temperature dependent. At 37 degrees C, a maximum was reached at 8 h. It was also found to be a saturable process with respect to homogenate and hormone concentrations. Association constants and number of binding sites determined in 9 men, using Scatchard plot and saturation curve analysis ranged, respectively, from 0.2 to 1.8 x 10(10) M-1 and from 92 to 3427 fmol/g testis or 7 to 380 fmol/mg protein. Degradation of [125I]hCG increased with temperature and time of exposure to human testicular homogenate. It increased also with increasing human testicular homogenate concentration and substrate concentrations. For a similar concentration of [125I]hCG, per cent of degraded hormone ranged from 32 to 57, according to the subjects. These results show that human testicular homogenates are capable of binding and degrading hCG in vitro. Biological and physiological implications of degradation for hormone binding are discussed. PMID- 6291299 TI - Specific binding sites for epidermal growth factor and its effect on human chorionic gonadotrophin secretion by cultured tumour cell lines: comparison between trophoblastic and non-trophoblastic cells. AB - Using human trophoblastic (SCH) and nontrophoblastic (HeLa S3) tumour cell lines, specific binding sites for epidermal growth factor (EGF), a potent stimulator of growth in many tissues, and its effect on secretion of human chorionic gonadotrophin (hCG) and/or its subunits were compared between these two tumour cells. Both SCH and HeLa S3 cells possessed two populations of specific binding sites for 125I-labelled EGF: the high affinity (Kd approximately 10(-10) M) and the low affinity (Kd approximately 7 x 10(-10) M) system. Tetradecanoyl phorbol acetate (TPA), a tumour promotor, showed a potent competitor of labelled tracer binding to its receptor sites in both cell lines. EGF stimulated both hCG-alpha and hCG and/or hCG-beta secretion in a dose-responsive manner from SCH cells, whereas it had no effect on hCG-alpha secretion from HeLa S3 cells. In contrast, dibutyryl cyclic AMP plus theophylline, a phosphodiesterase inhibitor, enhanced hCG-alpha secretion from both cells, while TPA had no effect in either cells. These data suggest that EGF may play a physiological role in hCG secretion from trophoblastic tissues and that the mechanism by which hCG and/or its subunits are secreted may differ between trophoblastic and non-trophoblastic tumour cells. PMID- 6291301 TI - A case report of pleuropulmonary Waldenstrom's macroglobulinemia associated with hepatocellular carcinoma. PMID- 6291302 TI - Regulation by phosphorylase kinase of phosphoprotein phosphatase activity: simultaneous control of protein phosphorylation and dephosphorylation in skeletal muscle. AB - Phosphorylase kinase from rabbit skeletal muscle inhibited the dephosphorylation of phosphorylase a by phosphoprotein phosphatase. Phosphorylation (activation) of phosphorylase kinase by cyclic AMP-dependent protein kinase greatly increased this inhibitory effect. Thus, phosphoprotein phosphatase is inhibited by phosphorylase kinase in a reversible manner (Gergely et al. (1976) Biochim. Biophys. Acta 429 809-816). In this paper the regulation by phosphorylase kinase at phosphoprotein phosphatase activity in different fractions of muscle extract and in the presence of various ligands has been investigated. The presence of phosphorylase kinase also affected the ligand control of phosphatase activity. Phosphorylase kinase almost cancelled the inhibitory effect of AMP but hardly influenced the activating effect of glucose, glucose 6-phosphate and caffeine. Calmodulin, glycogen and phosphorylase b (effectors of phosphorylase kinase) did not influence the inhibitory effect of phosphorylase kinase. Fractions of muscle extract also demonstrated the regulatory role of phosphorylase kinase. These fractions contained considerable amounts of phosphorylase kinase and phosphatase. Phosphatase activity was inhibited by phosphorylation reactions triggered by Mg++ and ATP. Heat-stable inhibitors were absent from these fractions, therefore the transient inhibition of phosphatase could be attributed to the phosphorylation of endogenous phosphorylase kinase. The introduction between phosphorylase kinase and phosphatase resulted in a loss of AMP sensitivity, i.e. AMP did not inhibit the activity of phosphatase in those fractions. Our results imply that the phosphorylation of phosphorylase kinase is equally important both in the formation of enzymatically active phosphorylase a and in the inhibition of dephosphorylation of phosphorylase a. The consequence of these two effects is the elevated level of phosphorylase a. PMID- 6291303 TI - The role of folate deficiency in electrophysiological neuropathy of experimental hepatoma of the rats. AB - Experimental hepatoma of the rats was induced by chronic administration of 3' methyl-dimethylamino-azobenzene. The folate level of the hepatoma-bearing rats tended to be low as compared with that of normal controls, and the co-existence of folate deficiency and reduced motor nerve conduction velocity of the dorsal nerve trunks appeared not uncommon. The hepatoma-bearing rats revealed the disturbance of serine to glycine conversion in serum, and folate administration to them prevented electrophysiological neuropathy along with normalization of serine to glycine conversion. Therefore, the reduced motor nerve conduction velocity could be at least in part the result of a metabolic impairment due to folate deficiency. PMID- 6291304 TI - Role of alpha-2 adrenergic affinity in the action of a non-sedative antispasticity agent. AB - Muscle hypertonus of central origin can be effectively reversed by either dopamine agonists or alpha-adrenergic antagonists. Because of its efficacy in reversing reserpine rigidity (a syndrome resembling Parkinsonism), SKF-7265 was examined to determine whether its action was mediated through alpha-adrenergic or dopaminergic receptors. The pharmacologic blocking activity of SKF-7265 was assessed by measuring blockade of the cardiovascular agonist responses induced by norepinephrine, epinephrine, isoproterenol, acetylcholine and histamine. The binding affinity of SKF-7265 was determined by displacement of 3H-spiperone from rat corpus striatum tissue and 3H-clonidine and 3H-WB-4101 displacement from rat cerebral cortical tissue. Using the cardiovascular responses, SKF-7265 was devoid of beta-adrenergic or cholinergic blocking effects and did not produce any behavioral or reflex deficits in awake animals. Receptor binding studies showed that SKF-7265 had equal affinity for alpha-1 and alpha-2 adrenergic receptors and little affinity for dopamine receptors. It is concluded that the efficacy which has been reported for the SKF-7265 induced reversal of reserpine rigidity and its potential value as an antispasticity agent may be attributed to its relatively high affinity for alpha-2 adrenergic receptors. PMID- 6291305 TI - Connatal polyneuropathy -- a case with proliferated microfilaments in Schwann cells. AB - A case of connatal polyneuropathy is described in a boy who died of pneumonia at the age of 2 years, and from whom sural nerve biopsies had been taken when he was 4 and 16 months old. Clinically, his disease was characterized by motor weakness and muscular flaccidity in the presence of normal intellectual development. The evolution of the connatal peripheral nerve lesion could be followed from the age of 4 months to death: The first biopsy evidenced the most serious pathologic changes. The findings were reminiscent of those encountered in a fetal nerve at 18 weeks of gestation. Furthermore, it showed numerous filamentous inclusions in Schwann cells. The second biopsy showed a sparsely myelinated nerve with bands of basement membrane apparently unrelated to cells arranged around the nerve's fibers. A few Schwann cells containing filamentous inclusions were still present. At autopsy, the findings were identical to those of the second biopsy. The possibility that this patient was transitionally exposed to a neurotoxic agent during pregnancy and that the biopsy findings represent a lesion that is still florid in the first and in a residual state in the second biopsy is considered. PMID- 6291306 TI - Sellar teratoma with melanotic progonoma. A case report. PMID- 6291307 TI - Mixed chemodectoma-ganglioneuroma of the conus medullaris region. AB - Report of a 33-year-old man with a year-long history of low-back pain radiating into the left leg. Neurologic examination upon admission to the hospital revealed a spinal compression syndrome at the level L5. Laminectomy at L 4/5 revealed an encapsulated intradural tumor measuring 4 X 2.5 X 2.5 cm. The tumor was attached to the dorsal root L 4 and extended downward to the conus region. Light and electron microscopy revealed features mostly consistent with a mixed chemodectoma and ganglioneuroma. PMID- 6291308 TI - Hypertrophic neuropathy in Sjogren;s syndrome. PMID- 6291309 TI - Electron microscopy of aminergic retinal neurons. AB - The synaptic connections of the dopaminergic retinal neurons of Cynomolgus monkeys and rabbits were studied in the electron microscope. In the Cynomolgus monkeys, a labelling method with 5,6-dihydroxytryptamine was used, whereas an autoradiographic technique with a monoamine oxidase inhibitor and (3H)-dopamine was used in the rabbits. The labelling method with 5,6-dihydroxytryptamine altered the normal ultrastructure of the dopaminergic neurons, but their synaptic structures were still easily recognized. The autoradiographic method left the ultrastructure of the labelled neurons unchanged, and large dense-cored vesicles could therefore be seen to be associated with the dopaminergic neurons in the rabbits. Other morphological characteristics could also be seen. The distribution of dopaminergic perikarya in the inner nuclear layer and dopaminergic processes in the inner plexiform layer was consistent with that of amacrine cells in both Cynomolgus monkeys and rabbits. Furthermore, the dopaminergic processes had output synapses of the conventional kind characteristic of amacrine cells. In both animals, the dopaminergic processes were presynaptic to amacrine cell bodies and both pre- and postsynaptic to other amacrine cell processes. The number of output synapses was larger than the number of input synapses. Serial and convergent synaptic arrangements were common. The synaptic organization of the indoleamine-accumulating neurons in the retinae of rabbits, cats, goldfish and carp were also investigated in the electron microscope. The neurons were labelled with 5,6-dihydroxytryptamine after the otherwise interfering dopaminergic processes had been destroyed. The indoleamine-accumulating neurons of the three species had a distribution of perikarya and processes like that of amacrine cells. They also had synapses of the conventional kind. In the rabbit and the cat, the indoleamine-accumulating processes had mainly reciprocal synapses with rod bipolar terminals in the innermost part of the inner plexiform layer. They were, however, also also pre- and postsynaptic to other amacrine cells, and in the cat, they had synaptic contacts with cone bipolar cells and the dendrites of ganglion cells. The indoleamine-accumulating neurons of the goldfish and carp were mainly pre- and postsynaptic to other amacrine cells, even if contacts with bipolar terminals were also seen. In the retinae of goldfish and carp, a subset of indoleamine-accumulating processes were discovered in the middle part of the inner plexiform layer that had a previously not recognized type of conventional output synapse with two postsynaptic elements. This synapse was called a branched conventional synapse. It was found and investigated in the retinae of normal goldfish and carp. Its presynaptic structures were found to be similar to those seen in certain conventional synapses with presynaptic "globular clusters" and only one postsynaptic element. PMID- 6291310 TI - Signs of activity and progression in chronic glaucoma. PMID- 6291311 TI - Typologia histologica WHO tumorum oculi eiusque adnexorum, Latine reddita numerisque disposita. A Latin translation and coded compendium of the World Health Organization's Histological typing of tumours of the eye and its adnexa. PMID- 6291312 TI - Specular microscopic investigations on the corneal endothelium and its involvement in corneal oedema. PMID- 6291313 TI - Treatment of Cushing's disease in children. PMID- 6291314 TI - Primary and secondary maternal cytomegalovirus infections and their relation to congenital infection. Analysis of maternal sera. AB - 4 382 new mothers were examined retrospectively with the enzyme-linked immunosorbent assay (ELISA) for IgG activity to cytomegalovirus (CMV) during pregnancy. Some of them were also studied with the indirect immunofluorescence (IIF) test for CMV-IgM antibodies. All the infants had been studied for CMV excretion within the first week of life. Nineteen of them had been shown to be congenitally infected with CMV. 1 218 (28%) women lacked CMV-IgG activity at their first antenatal visit (usually in months III-IV). Fourteen of them seroconverted before parturition (primary infection). Thirteen of the seroconverters were shown to develop CMV-IgM activity. In 6 (43%) cases the primary infection was transmitted to the offspring. The remaining 13 congenitally infected infants were born to mothers with a positive IgM-test at their first antenatal control. Only one of these mothers had a clearly positive IgM-test. She was shown to lack CMV-antibodies before conception (primary infection during the first trimester). Preconceptional sera were obtained from further 4 of the 13 seropositive mothers of congenitally infected infants; all 4 had CMV antibodies before pregnancy (secondary infection during pregnancy). The combined studies of the mothers and infants revealed that 21-63% of the congenital infections could have been caused by secondary maternal infections. Prospectively performed, the study would only have disclosed one of the three fetal CMV infections that resulted in neurological sequelae. PMID- 6291315 TI - Pituitary-adrenal and testicular function in preterm infants after prenatal dexamethasone treatment. AB - Pituitary-adrenal and testicular function was monitored by plasma ACTH and testosterone (T) measurements in preterm newborns (gestational age below 36 weeks), exposed in utero to dexamethasone treatment for prevention of respiratory distress syndrome. A group of age-matched premature newborns and full-term infants served as controls. The cord-blood ACTH level was high in each group (logarithmic means 65-75 ng/l), but decreased within the first 2 days of life to mean levels between 20-30 ng/l on days 3 to 10 inclusive. Dexamethasone treatment had no effect on the postnatal ACTH concentrations when compared with preterm or full-term controls. Similarly, no difference in plasma ACTH were found between untreated preterm and full-term infants during the first 10 days of life. T levels in mixed cord blood were on average 2-3 nmol/l in the preterm male infants. An increase to a mean level of 10 nmol/l was seen in 1 h and 1 d samples. Thereafter, the concentration of T decreased to 2-3 nmol/l on days 3-10 postnatally. No effect of dexamethasone treatment was seen on the postnatal pattern of plasma T. When preterm male and full-term male infants were compared, no difference was seen in the T peak in the first day of life. However, the 1-3, and 60-90 days concentrations of T were 2-fold (p less than 0.01-0.05) higher in the preterm group. It is concluded that prenatal dexamethasone treatment of the mother does not influence the postnatal pituitary-adrenal function as monitored by ACTH measurements. Likewise, no effect of this treatment was observed on the postnatal testicular activity of preterm male infants. The immediate postnatal peak in plasma T levels persisted longer in the preterm than in the full-term male infants. PMID- 6291317 TI - Ganglioneuroblastoma containing several kinds of neuronal peptides with watery diarrhea syndrome. AB - This report presents an adrenal ganglioneuroblastoma containing several kinds of neuronal peptides. The tumour was found in the autopsy case of a 3-year-old girl with clinical manifestation of intractable diarrhea, hypokalemia, achlorhydria, and with elevated levels of plasma vasoactive intestinal peptide (VIP). Immunoperoxidase staining showed many immunoreactive VIP- containing cells, some somatostatin-and substance P-containing cells on the tumour sections. Ultrastructurally, the tumour cells contained numerous secretory granules that could be divided mainly two types; one is a small cored vesicle (50-150 nm in diameter) and the other large electron dense secretory granule (200-500 nm in diameter). It was suggested that the cells in ganglioneuroblastoma derived from neural crest are closely related to the cells that could differentiate into gut hormone-producing cells. PMID- 6291316 TI - Vitamin D metabolism in hypophosphatasia. AB - A 4-month-old boy with the infantile form of hypophosphatasia was followed for 9 months with measurements of serum calcium, phosphate, alkaline phosphatase and various vitamin D metabolites, together with urinary excretion of cyclic AMP. During the initial hypercalcemic stage the serum concentration of 25 hydroxyvitamin D was normal. Urinary cyclic AMP was low and the serum concentration of the dihydroxymetabolites of vitamin D were appropriate to the high serum calcium with low 1,25-(OH)2D and relatively high 24,25(OH)2D and 25,26(OH)2D levels. Due to restrictions of the vitamin D intake and lack of exposure to sun he developed vitamin D-deficiency rickets at 9 months of age with very low serum concentration of 25-hydroxyvitamin D and markedly increased urinary excretion of cyclic AMP. Following vitamin D treatment the serum level of 1,25(OH)2D showed a brisk rise to a considerably elevated value. Initially the serum concentration of alkaline phosphatase was well below the normal range, rose markedly during the stage of active rickets and returned to the characteristic low levels of hypophosphatasia with healing of the rickets. PMID- 6291318 TI - Inhibition of human lung cyclic nucleotide phosphodiesterases by proxyphylline, theophylline and their metabolites. AB - Inhibition of cAMP and cGMP hydrolysis in human lung tissue by proxyphylline and its main metabolite 1-methyl-7-(beta-hydroxypropyl)xanthine was compared with theophylline and its metabolite 3-methylxanthine. The apparent inhibition constant of proxyphylline was 0.06-0.7 mmol/l at low cAMP concentrations and at low and high cGMP concentrations, while it was 1.0 mmol/l at high cAMP concentrations. This is 1.2-1.7 times higher than the corresponding values for theophylline. Both metabolites were slightly weaker inhibitors than their parent drugs, the theophylline metabolite relatively weaker than the proxyphylline metabolite. The differences in the observed inhibition constants agree with previous clinical studies indicating that the potency of theophylline is approximately twice that of proxyphylline. PMID- 6291319 TI - The need of protein synthesis for the effect of LH--RH on the cAMP level in the anterior pituitary in vitro. AB - The effect of LH--RH on cAMP accumulation in the anterior pituitary of male rats was investigated. The effect consisted of two phases. In the first phase of incubation (about 2 hours), there was no change in the cAMP level, while in the second phase a significant increase was observed. Cycloheximide (10(-5)M) given simultaneously with the LH--RH totally blocked the observed effect of LH--RH. If cycloheximide was given in the second phase of incubation, when the cAMP level was already elevated, its further rise was prevented and the elevated cAMP level remained unchanged for 2 or more hours. Actinomycin D (10(-5)M) added together with LH--RH totally abolished the action of the latter, but if actinomycin D was given 1 hour after LH--RH a significant increase of the cAMP level was found at the end of the 4-hour incubation. PMID- 6291320 TI - Structure-activity studies concerning effect of cholecystokinin octapeptide on passive avoidance behaviour of rats. PMID- 6291321 TI - Effects of ACTH and PGE2 on ovarian venous outflow, progesterone and oestradiol 17 beta secretion in the HCG primed bitch. PMID- 6291322 TI - Modulation of passive avoidance behaviour of rats by intracerebroventricular administration of cholecystokinin octapeptide sulfate ester and nonsulfated cholecystokinin octapeptide. AB - The effects of intracerebroventricular administration of different doses of cholecystokinin octapeptide sulfate ester (CCK-8-SE) and nonsulfated cholecystokinin octapeptide (CCK-8-NS) were tested on the latency of passive avoidance behaviour in rats. Treatments were carried out prior to learning trial, immediately after electroshock and prior to testing 24 h retention. Both CCK-8-NS and CCK-8-SE enhanced the latency of passive avoidance after all forms of treatment while showing different dose-response patterns depending on time of administration. These data indicate that CCK-8-SE and CCK-8-NS might play a role in the regulation of memory consolidation and retrieval. PMID- 6291323 TI - Activation of NMDA-receptors elicits "fictive locomotion" in lamprey spinal cord in vitro. PMID- 6291324 TI - Modulation by cholecystokinins of 3H-spiroperidol binding in rat striatum: evidence for increased affinity and reduction in the number of binding sites. PMID- 6291325 TI - Metabolic and hormonal adjustments during hemorrhage in cats after interference with the sympatho-adrenal system. AB - The relative contribution of the splanchnic sympathetic innervation and the adrenal medulla for metabolism and hormone secretion during two different levels of hemorrhagic hypotension was investigated in 3 groups of anesthetized cats, viz, intact, adrenalectomized and splanchnicotomized (adrenalectomy + cutting of splanchnic nerves). In intact cats, hemorrhage caused very marked elevations of arterial plasma glucose, adrenaline, noradrenaline, dopamine, lactate, cAMP, glycerol and glucagon concentrations whereas plasma insulin fell to only 20% of control values. Adrenalectomy attenuated the glucose, adrenaline, noradrenaline and cAMP responses whereas the normal insulin inhibition was abolished. Splanchnicotomy further reduced the hemorrhagic glucose and glycerol responses and, possibly, also that of glucagon. It is concluded that the adrenergic system as a whole is important for the adjustments of the release of glucose, cAMP, glycerol, insulin and glucagon that occur during hemorrhage in cats. The adrenal medulla seems to be of particular importance for the regulation of cAMP release. PMID- 6291326 TI - VIP-induced cyclic AMP formation in the cat submandibular gland. Potentiation by carbacholine. PMID- 6291327 TI - Electronic properties of motor nerve terminals. AB - To obtain information about the electric membrane properties of frog motor nerve terminals we examined how depolarizing or hyperpolarizing current pulses of 2-8 ms duration to the preterminal, by electrotonic spread of potential, affected depolarization induced transmitter release. Sodium channels were blocked by tetrodotoxin. Under this condition a hyperpolarizing current pulse produced inhibition of release, followed by potentiation of release. Inhibition lasted more than 100 ms with a time constant of 50-150 ms. When, in addition, potassium channels were blocked by 3,4-diaminopyridine or tetraethylammonium a depolarizing current pulse potentiated transmitter release for a period up to 50 ms. The results imply that inward currents in the nerve terminal are carried mainly by sodium and calcium ions and outward currents by potassium ions while "leak" conductances are negligible. A low "leak" conductance and therefore a high specific membrane resistance facilitates the spread of electronic potentials and thereby explains the longlasting effects on transmitter release of brief current pulses to the preterminal. PMID- 6291328 TI - Neural beta-adrenergic dilatation of the facial vein in man. Possible mechanism in emotional blushing. AB - Ring preparations of the superficial buccal segment of the human facial vein, taken from extirpated tissue in 12 patients during neck surgery, were studied in vitro. The vein developed a maintained intrinsic myogenic tone in response to passive stretch and was supplied with alpha- as well as beta-adrenoceptors, both of which could be influenced by transmural nerve stimulation (TNS) and noradrenaline. These unusual characteristics for a vein are basically similar to the ones described for the rabbit facial vein by Pegram, Bevan & Bevan (1976). In man there seemed to be an inter-individual difference with regard to the abundance of 'innervated' alpha- and beta-adrenoceptors. Facial vein specimens from some subjects thus responded with prompt and pronounced net dilatation to TNS with maximum at 4 Hz and those from others with net constriction with maximum at 16 Hz. The latter showed a reversal into neural beta-adrenergic dilatation after alpha-adrenergic blockade. The human external jugular vein was devoid of intrinsic tone and beta-adrenoceptors. It is tentatively proposed that a beta adrenergic neuro-effector mechanism in superficial ramifications of the facial vein in man might be involved in the emotional blushing reaction. PMID- 6291329 TI - Exchange of isethionate between blood and tissues in adult and 7-day-old mice. AB - [14C] Isethionate was injected intramuscularly into adult and 7-day-old mice and the distribution of the label in the blood, brain, heart, liver and spleen was determined. Endogenous isethionate in these tissues was determined and approximate exchange rates calculated from the endogenous isethionate tissue levels and specific radioactivities after certain time intervals. The concentration of isethionate was higher and its exchange rates between plasma and tissues faster in the adult mice than in the 7-day-old mice. A close correlation between isethionate transport rates and tissue levels in vivo was obtained in both age groups. The tissues eliminated isethionate rapidly, even faster in the adult than in the 7-day-old mice. The elimination of isethionate showed three different components, fast, intermediate and slow, in the heart, liver and spleen of the adult mice, but only two components, fast and slow, in the other cases. PMID- 6291331 TI - Inhibition of gastric acid secretion inthe Atlantic cod, Gadus morhua, by sulphated and desulphated gastrin, caerulein, and CCK-octapeptide. AB - Gastric acid secretory effects of gastrin/CCK-like peptides have been assayed in cods rendered "spontaneously" secreting by a continuous intestinal perfusion with diluted (33%) seawater. A high dose of pentagastrin induced a weak stimulation (31%) of acid output, while gastrin 17-II, caerulein and CCK8 were inhibitory. Caerulein was the most potent peptide, with an estimated D50 for inhibition of 0.013 nmol/kg.h. Although displaying lower potencies, also the desulphated forms of gastrin-17, caerulein and CCK8 were inhibitory. The results may be explained by release of an endogenous inhibitor, or by interaction with endogenous "codfish gastrin". In the latter case two alternatives are considered: Either gastrin 17, CCK8, and caerulein possess lower efficacies than "codfish gastrin" and therefore act as partial agonists. Alternatively, "codfish gastrin" is itself an inhibitory principle (gastron), the effect of which is mimicked by gastrin 17, caerulein and CCK8. The actions of gastrin and the gastrin-like peptides in the cod indicate a structure-activity relationship different from previously described systems, both mammalian and submammalian. PMID- 6291330 TI - Quantitative pharmacological characterization of beta-receptors and two types of alpha-receptors mediating sympathomimetic smooth muscle response in the human Fallopian tube at various cyclic stages. AB - The dissociation constants for adrenoceptor-antagonist complexes (KB) were determined in vitro in circular and longitudinal smooth musculature from the ampullary and isthmic regions of the human Fallopian tube. High extracellular potassium concentrations were used to eliminate the spontaneous contractile activity. Neuronal and extraneuronal amine uptake mechanisms were blocked. The parallel shift of the log dose-response curves was secured in Arunlakshana-Schild plots. KB for the beta-receptor, mediating sympathomimetic relaxation, were determined during alpha-receptor blockade: the values for propranolol were the same (approximately 10(-6) M) in all preparations and at all cyclic stages, as determined from plasma estradiol and progesterone levels. KB for the complex between the alpha-receptor (mediating contraction) and phentolamine were determined during beta-receptor blockade. The values were the same in all types of smooth musculature, but varied with cyclic stage: they were around 7 x 10(-8) M when plasma estradiol and progesterone were both minimum, and around 2 x 10(-7) M when these steroid levels were moderate to high, suggesting that the properties of the contractile receptors of the human Fallopian tube are modified during the menstrual cycle. PMID- 6291332 TI - Evidence for stabilization of cortical 5HT neurotransmission by chronic treatment with antidepressant drugs: induction of a high and low affinity component in 3H 5HT binding sites. PMID- 6291333 TI - Effect of stress and dexamethasone on immunoreactive beta-endorphin levels in rat hypothalamus and pineal. AB - In response to mild stress the levels of immunoreactive beta-endorphin in rat anterior pituitary, hypothalamus and pineal fell within 10 minutes from 210 to 129 pmol/lobe, 1.47 to 0.89 pmol/mg protein and 2.53 to 0.41 pmol/gland, respectively. No alterations were found to take place in beta-endorphin levels in posterior pituitary or plasma. Dexamethasone pretreatment given 18 h prior to stress resulted in significantly greater reduction of beta-endorphin levels in hypothalamus and pineal than stress alone--hypothalamic levels fell to 0.73 pmol/mg protein and pineal to 0.07 pmol/gland. Plasma beta-endorphin levels in dexamethasone pretreated stressed rats were significantly lower than in intact rats (42 fmol/ml vs. 98 fmol/ml). The almost complete disappearance of beta endorphin from the pineal in response to stress and dexamethasone suggests that pineal does not itself synthesize the hormone but only utilizes and/or stores it. Gel filtration analysis of the beta-endorphin immunoreactivity in tissue extracts and plasma showed that anterior pituitary and plasma contain three immunoreactive components, eluting like beta-endorphin, beta-lipotropin and proopiocortin, whereas only beta-endorphin-like material was detected in posterior pituitary, hypothalamus and pineal. PMID- 6291334 TI - Adrenergic responses in different sections of rat airways. AB - Effects of adrenergic agonists on 4 different preparations (trachea, bronchus, bronchiole and parenchymal strip) of rat airway smooth muscle were studied. Phenylephrine, an alpha-agonist, contracted the trachea, bronchus and parenchymal strip but had no effect on the bronchiole. Similarly adrenaline, both a beta- and an alpha-agonist, contracted the trachea, bronchus and parenchymal strip, but neither contracted nor relaxed the bronchiole. These results suggest that the rat trachea and bronchus contain alpha receptors but the bronchiole does not. Alpha response of the parenchymal strip preparation seems to originate from the vascular smooth muscle. Isoprenaline, a nonselective beta-agonist, could not elicit any response on resting airways, but caused a relaxation of all four preparations after being contracted by carbacholine. Expressed in % of maximal carbacholine contractions, the relaxations were: 21% in the trachea, 45% in the bronchus, 92% in the bronchiole and 141% in the parenchymal strip. This shows that beta response is weak in the large central airways but becomes stronger towards the small peripheral airways, when measured against cholinergic contraction. PMID- 6291335 TI - Adenosine actions and adenosine receptors after 1 week treatment with caffeine. AB - After one week treatment with caffeine (20 mg/kg i.p.) the number of adenosine receptors, as determined by specific binding of (3H)-L-PIA, in rat cerebral cortical membranes was increased by about 25%. Cyclic AMP accumulation induced by adenosine analogues in slices of rat hippocampus was unaffected by caffeine treatment. The inhibition of lipolysis in rat fat cells by 2-chloro-adenosine was similarly unaffected. The potency of caffeine as an antagonist of these adenosine receptor mediated effects was not altered by caffeine treatment. It is concluded that at least some adenosine receptors are up-regulated as a consequence of prolonged caffeine treatment, but that the increase in receptor number is not related to changes in at least two effects of adenosine and caffeine. PMID- 6291337 TI - Sheep-pox vaccine prepared from formaldehyde inactivated virus adsorbed to aluminium hydroxide gel. AB - The aim of the study was to produce an efficacious formaldehyde inactivated, adsorbed vaccine from the Mongolian sheep-pox MH virus strain. Aluminium hydroxide gel prepared from AlCl3 X 6 H2O proved to be the most efficacious adsorbent among the gels prepared from different substances. Above 1.8% Al2O3 content the unadsorbed virus quantity was less than 1% of the original one. Using gel prepared from KAl(SO4)2 and AlCl3 X 6 H2O, respectively, the quantity of adsorbed virus was the same during the adsorption period from 10 min to 24 h. Intradermal inoculation of sheep proved more advantageous for virus production than subcutaneous inoculation. Three vaccines containing different quantities of antigen were prepared from virus propagated in sheep. The vaccine containing 19 800 ID50 inactivated virus did not protect the sheep even against a virus challenge of 25 ID50, while that of 67 000 ID50 content protected 50% of the animals infected with 125 to 287 ID50, and that of 395 000 ID50 content protected 100% of the animals against challenge with more than 100 000 ID50. More than 3 million sheep were inoculated in Mongolia with vaccines of 350 000 ID50 virus content in the last years. In the areas where vaccination has been introduced no sheep-pox epizootic has occurred. PMID- 6291338 TI - In-vitro secretion of ACTH in Nelson's syndrome. AB - Cell suspensions of ACTH cell adenomas of 10 patients with Nelson's syndrome were investigated for in-vitro secretion of ACTH. Two incubation systems, one using incubation beakers and the other a superfusion system, were employed. The cells were tested for their reactivity to lysine-vasopressin (LVP), cortisol, and combinations of both. LVP regularly provoked a rapid significant increase of ACTH secretion. The effect of cortisol was heterogeneous. Paradoxical initial stimulatory effects of cortisol were observed. There was a suppressive effect in some patients, which correlates to low proliferative activity in histological evaluation. In both systems a low secretory activity coincided with high proliferative activity in vivo. PMID- 6291336 TI - Regional difference in cyclic AMP response to adenosine of rat cerebral cortex with an iron-induced epileptic focus. AB - A chronic epileptic focus not resulting in generalized convulsions was induced by a microinjection of FeCl3 solution into the left anterior cortex of rats. Cyclic AMP accumulation in response to adenosine was examined in incubated slices from the cerebral cortex of animals that showed bilateral spike and slow wave complex in an electrocorticogram (ECoG) 30 to 60 days after the microinjection. Cyclic AMP accumulation after incubation with adenosine was most marked in slices from the left anterior quadrant of the cortex including a FeCl3-injected site. A medium response was observed in both the left posterior and the right anterior quadrants, but little or no response in the right posterior quadrant of the cortex. PMID- 6291339 TI - Pituitary function before and after transsphenoidal adenomectomy in patients with Cushing's disease. AB - Over the past 4 years 11 patients with Cushing's disease have been operated on by a transsphenoidal microsurgical technique. All patients had small intrahypophyseal adenomas found at operation. Eight of the 11 patients had a remission of the disease on the basis of clinical evaluation and hormonal studies. The endocrinological results revealed a transient state of hypocorticism in all successfully treated patients. Within 6-8 months after operation recovery of hypothalamic-pituitary-adrenal function and other tropin function took place. These findings, together with the absence of recurrence up to now, further confirm the efficacy of transsphenoidal surgery in the management of patients with Cushing's disease. PMID- 6291340 TI - Evaluation of surgically treated Nelson's syndrome. AB - In 17 primary transnasal operations on Nelson tumours long-term results are assessed measuring ACTH plasma levels and performing regular neuroradiological controls. In small intrasellar adenomas only one recurrent ACTH elevation near to pretreatment levels without symptoms from tumour growth was observed after 10 years. ACTH normalization was rarely achieved in large tumours, but only in one patient was a relapse visible in CT scan after three years. Thus selective tumour removal seems to be advisable at an early stage when sellar enlargement occurs. Interpretation of ACTH plasma levels has to include completeness of adrenalectomy, cystic tumour necrosis, and corticoid replacement which reduces ACTH levels. Elevation of ACTH plasma levels over 5,000 pg/ml has been regularly accompanied by extrasellar tumour extension. In large adenomas with definitely elevated ACTH levels, additional radiation therapy should be considered. PMID- 6291341 TI - Decompressive frontal lobe resections and brain oedema. PMID- 6291342 TI - Receptors, receptor sensitivity, and receptor regulation in the CNS. PMID- 6291343 TI - Studies on the cell association of exogenous glycolipids. PMID- 6291344 TI - Reduced ambient temperature blocks the ability of naloxone to prevent endotoxin induced hypotension. PMID- 6291345 TI - The pathophysiology of septic shock: acute renal failure in rats following live E coli injection. A histochemical study of the proximal tubules. AB - Acute renal failure was induced in rats by injection of a lethal dose of live Escherichia coli. Enzyme activities of the proximal tubule were studied histochemically at three, six, and 12 hours following E coli injection. The enzymes examined were alkaline phosphatase (A1Pase), acid phosphatase (AcPase), adenosine triphosphatase (ATPase), succinate dehydrogenase (SDH), glucose-6 phosphatase (G6Pase), and glucose-6-phosphate dehydrogenase (G6PDH). At three hours, ATPase activity was slightly decreased, while other enzymes showed no changes in activities at this time. At six hours, a slight increase in AcPase activity was seen in the pars recta. At this time, although A1Pase showed no change in activity, other enzymes revealed slight decreases in activities: G6Pase and SDH in the pars convoluta, ATPase in the pars convoluta and pars recta, and G6PDH in pars recta. At 12 hours after treatment, all enzymes showed decreases in activities; however, no necrotic tubule changes were detectable by light microscopy. Since sodium reabsorption in proximal tubules requires a sodium pump consisting of Na-K ATPase, early histochemical changes in ATPase activity in proximal tubule following bacteremia may be related to early changes in sodium reabsorption causing polyuria and to the subsequent development of acute renal failure. PMID- 6291346 TI - Studies of the mechanism of hypoxic pulmonary vasoconstriction. AB - The intrapulmonary mechanism by which airway hypoxia causes pulmonary arterial constriction is poorly understood. It is generally believed that hypoxia either elicits the release of a chemical mediator from the lung parenchyma or has a direct excitatory effect on the smooth muscle of the peripheral pulmonary arteries. We are testing the working hypothesis that hypoxia acts directly on the vascular smooth muscle to depress the rate of mitochondrial oxidative phosphorylation and to cause shifts in cytoplasmic metabolite concentrations, which then lead to membrane depolarization, calcium influx, and contraction. Results from studies of isolated perfused rat lungs with pharmacologic inhibitors of oxidative phosphorylation, glycolysis, and calcium influx have provided indirect support for the hypothesis; but a simpler in vitro preparation allowing direct measurements of energy metabolism, membrane electrical activity, calcium fluxes, and muscle tone is needed. Additional experiments have shown that inhibitors of membrane K+ conductance allow hypoxic contractions of isolated vascular smooth muscle; and such a preparation might be useful as an in vitro model of hypoxic pulmonary vasoconstriction. PMID- 6291347 TI - The effects of clofibrate ingestion on alveolar macrophage peroxisome content and oxygen metabolism. AB - Respiratory burst activity in alveolar macrophages in response to particulate and soluble challenges, such as zymosan particles and phorbol myristate acetate (PMA), is not nearly as dependent upon membrane stimulation as in neutrophils. Microperoxisomes are subcellular organelles containing catalase and are present in lung macrophages and cells of other organs. Evidence from liver cells indicates that peroxisomes are intimately involved with hydrogen peroxide and lipid metabolism. Clofibrate (2-(p-chlorophenoxy)-2-methylpropionic acid ethyl, Atromid-S-), a hypolipidemic drug known to cause peroxisomal proliferation in liver cells, was studied with respect to its ability to cause increases in the microperoxisome content and to alter the cellular metabolism of alveolar macrophages. Liver weight increased over a 2-week drug treatment period while lung weight remained unchanged. Plasma triglyceride levels were decreased by the treatment, indicating the effectiveness of the drug. Unlike the effect on liver cells, however, clofibrate did not cause a proliferation of microperoxisomes, as determined by morphometric analysis. Oxygen consumption and hydrogen peroxide generation by alveolar macrophages in response to either stimulant (zymosan or PMA) was no greater in clofibrate-treated rats than in controls. Superoxide release, when expressed as the change in response to PMA, appeared elevated in the drug group; statistical significance, however, was not demonstrated. The hexose monophosphate shunt (HMP), which produces reducing equivalents for lipid biosynthesis, was elevated in macrophages from clofibrate-treated rats when expressed similarly. The significance of these results in relation to the known effects of the drug on liver cells. PMID- 6291348 TI - Antioxidation theory of non-steroidal anti-inflammatory drugs based upon the inhibition of luminol-enhanced chemiluminescence from the myeloperoxidase reaction. AB - The action of non-steroidal anti-inflammatory drugs (NSAIDS) has been ascribed to their ability to block the reaction of arachidonate with cyclooxygenase/peroxidase, thus inhibiting the cellular production of inflammation mediators such as prostaglandins and leukotrienes. However, this and other polymorphonuclear leukocyte (PMN) peroxidases such as myeloperoxidase (MPO) would still be capable of producing destructive oxidants which contribute to inflammation. Sulindac sulfide (Clinoril sulfide) has recently been shown to scavenge oxidant products of prostaglandin cyclooxygenase/peroxidase and MPO. The MPO-H2O2-Cl- reaction is a potent antimicrobial/cytotoxic system which produces HOCl, a strong oxidant. MPO itself has the ability to oxidize drugs and cellular components, and may be the main oxidant in PMN defenses. An antioxidant/free radical scavenger action of NSAIDs against the MPO system could be a primary mechanism of their anti-inflammatory effects. Other antioxidant/free radical scavengers have anti-inflammatory effects. MPO activity has previously been quantified using chemiluminescence (CL). In this study, NSAIDs from various classes were tested for their ability to inhibit luminol-enhanced CL from MPO. The most potent NSAIDs against MPO-CL were BW755C, phenylbutazone, indomethacin and sulindac sulfide. Salicylates and arylacetic acid derivatives, such as naproxen, also decreased MPO-CL. These drugs are also effective against CL from PMNs, of which MPO may be a main source. This effect of NSAIDs on MPO suggests that NSAIDs may impair the killing mechanism of the PMN, preventing cell destruction and release of inflammation mediators. PMN MPO appears to be a target for the antioxidant/free radical scavenging effects of NSAIDs. PMID- 6291350 TI - [Studies on cyclic nucleotides and protein kinase of lens. II. Endogenous substrate proteins for cyclic AMP dependent protein kinase in bovine lens]. PMID- 6291349 TI - Histamine as a ligand in blood plasma. Part 5. Computer simulated distribution of metal histamine complexes in normal blood plasma and discussion of the implications of a possible role of zinc and copper in histamine catabolism. AB - Previously physiological experiments carried out on mice have proved that copper and zinc can interfere with the pharmacological effects of histamine that lead to anaphylactic shock. A quantitative study of the interactions between essential metal ions and histamine in plasma was thus undertaken. The progressive approach towards a reliable computer-simulated distribution of the histamine-containing plasma species necessitated a large series of physicochemical determinations of the formation constants of the binary and ternary metal complexes involved. The present paper deals with the determination of the formation constants in the zinc serine, zinc-histamine-serine, zinc-histamine-lysine, copper-serine, copper histamine-serine, and copper-histamine-valine systems, which were still necessary to reach the reliable simulation required. The subsequent final distribution of histamine in plasma has thus been computed, and interpreted in terms of a possible role for zinc in assisting the histamine catabolism process. Further computer calculations simulating the increase of the zinc concentration in human blood plasma support this interpretation. The antagonizing role of copper against that of zinc has also been examined. PMID- 6291352 TI - Dilated cardiomyopathy in young adult Africans: a sequel to infections? AB - Over a period of 6 years, only twelve cases of dilated cardiomyopathy were clinically diagnosed in Nigerians between the ages of 11-30 years at the University College Hospital, Ibadan, Nigeria. Eleven presented with heart failure, while the twelfth patient presented with a cerebrovascular accident. Two other patients also had a cerebrovascular accident. A history of febrile illness was obtained in seven, but in only three was fever unresponsive to antimalarials, documented on admission. Antistreptolysin-O titre was normal and erythrocyte sedimentation rates elevated in each of the patients. Leucocytosis was present in six, three had a four-fold rise or fall in antibody titres against Coxsackie-B viruses and one, a four-fold rise or fall against Toxoplasma gondii. Histological evidences of myopericarditis were found in three of the six patients who died. It is concluded that dilated cardiomyopathy is rare in young adult Nigerians, and that constitutional upset is common, as in children, but prognosis is poorer. Infections by Coxsackie-B viruses, T. gondii and possibly other viruses appear to be of major aetiological factors PMID- 6291351 TI - Studies on interrelationship between calcium translocation and hydrogen carbonate secretion in the avian oviduct. PMID- 6291353 TI - Characteristics of boric acid tolerant Vibrio cholerae. AB - Two strains of boric acid tolerant Vibrio cholerae have been studied in order to identify the characteristics which distinguish them from the 'wild-type' virulent strains. Strain Ib5 requires either amino acid cysteine or methionine while strain Ib5S requires deoxyribonucleic acid (DNA) for growth. Although the strains did not grow on desoxycholate citrate agar (DCA) they grew well on chocolate agar. Morphologically filamentous, bacillary and comma-shaped forms were seen, depending on the media and incubation period. The strains failed to cause accumulation of fluid in gut ligation and permeability factor tests. With the single radial immunodiffusion test, strain Ib5S gave positive reaction in 83% of the tests when DNA was added to the growth medium. On the other hand only 25% of the tests were positive when no DNA was added to the growth medium, even though the antigen was concentrated ten-fold. Strain Ib5 only gave 50% positive tests when the antigen was concentrated ten-fold. It is therefore suggested that Ib5S strain is a cholerangenoid producer in the presence of DNA, and may be useful as an effective vaccine against cholera. PMID- 6291354 TI - Plasma cortisol responses to cortrosyn treatment in asthmatic children. AB - Treatment trial over a period of 3 months was conducted with intermittent intramuscular injections of Cortrosyn depot in fourteen children with severe and frequent asthmatic attacks. The basal plasma cortisol levels were generally high, but higher than normal in four (29%) of the patients. At a period of 24 h after the initial Cortrosyn injection was administered, plasma cortisol increases ranging between 4-52 micrograms/100 ml above the basal levels were recorded. At a period of 1 week after the end of daily injections for 1 week, increases of plasma cortisol ranging between 3-71 micrograms/100 ml above the basal levels were observed and presumed to be a reflection of an associated adrenal hypertrophy resulting from repetitive daily Cortrosyn injections. The highest increases at this stage were observed in the youngest patients with the severest asthmatic attacks, but not in their older counterparts. At the end of the trial treatment, clinical improvement was associated with lowered plasma cortisol levels compared with the elevated basal values. PMID- 6291355 TI - Field trials of pyrantel pamoate (Combantrin) in Ascaris, hookworm and Trichuris infections. AB - In a trial involving 185 school children, pyrantel pamoate (Combantrin) at a dose of 20 mg/kg body weight for 1-3 days was found to be very effective against the Ascaris and moderately effective against the hookworm, with mean cure rates ranging from 93.3-96.7% and 53.3-73.3% respectively. No apparent action against the Trichuris was detected, a mean cure rate of between 34.2 and 46.1% being only slightly, but not significantly, better than the 33.8% cure for a placebo-treated control group. Single and multiple doses of the suspension and tablet formulations of the drug were well tolerated. From the series of randomized and controlled trials conducted, we recommend that, in this area, the appropriate dose of the drug to use for treating ascariasis is 10 mg/kg per day for 1 day, and for infections which include hookworm, 20 mg/kg per day for 3 days. PMID- 6291356 TI - Rhinoentomophthoromycosis. PMID- 6291357 TI - Antipyrine kinetics in Nigerian women in chronic renal failure. AB - Antipyrine was given orally to five patients with Chronic renal failure and nine normal subjects. Plasma antipyrine levels were measured by high pressure liquid chromatography method, the plasma half-life of the drug was determined and used as an index of drug oxidation. The mean (+/- s.d.) plasma antipyrine half-life in patients with chronic renal failure (7.8 +/- 2.6 h) was significantly shorter than in normal subjects (13.1 +/- 2.3 h) (P less than 0.001). There was no significant difference in the volume of distribution but there was a significant difference in the clearance in ml.kg-1h-1 (P less than 0.001). The results suggest that oxidation of antipyrine by hepatic microsomal enzymes is increased in patients with chronic renal failure. The role of this in the apparent resistance among our patients to antihypertensive agents needs further study. PMID- 6291358 TI - Vasodilator therapy of severe congestive heart failure: the special importance of angiotensin-converting enzyme inhibition with captopril. AB - Successful ambulatory afterload reduction therapy of severe chronic congestive heart failure (CHF) required the extensive evaluation of hemodynamic effects of vasodilator agents, precise characterization of differential cardiocirculatory actions, and objective confirmation of extended salutary improvements of the heart failure state. This article describes results of a series of investigations with the angiotensin-converting enzyme (ACE) inhibitor captopril (CPT) in several patients with severe CHF. CPT causes predominant peripheral venodilation, resulting in marked decline in elevated left ventricular preload and modest augmentation of the depressed cardiac output. These hemodymanic effects of oral CPT are similar to the effects of nitroprusside and prazosin on increased ventricular preload, but the latter vasodilators cause greater rise in low cardiac pump output. Importantly, the beneficial cardiocirculatory action of oral CPT provided prolonged cardiac benefits with symptomatic improvements confirmed by objective enhancement in left ventricular function documented by cardiac catheterization, echocardiography, nuclear scintigraphy, and treadmill exercise. Thus, ACE inhibition with oral CPT successfully provides marked long-term augmentation of cardiac performance and clinical status in refractory CHF. PMID- 6291359 TI - Captopril in congestive heart failure: improved left ventricular function with decreased metabolic cost. AB - The oral angiotensin-converting enzyme inhibitor captopril (CPT) produces beneficial hemodynamic and clinical responses in patients with chronic congestive heart failure (CHF). Cardiac output and stroke volume increase, along with a decrease in pulmonary capillary wedge pressure, indicating improved left ventricular function. During maintenance CPT therapy, the beneficial hemodynamic and clinical effects appear to be sustained. Improved left ventricular pump function with CPT is associated with decreased metabolic cost, as myocardial oxygen consumption consistently decreases in proportion to the decrease in myocardial oxygen demand. Myocardial ischemia occurs infrequently, as is evident from the abnormal myocardial lactate metabolism. Hypotension appears to be the major adverse effect, particularly after the first dose. However, with dose titration and the use of a smaller initial dose, a marked precipitous fall in blood pressure can be avoided in the majority of patients. Thus, CPT may prove to be a useful agent in the vasodilator therapy of chronic CHF. PMID- 6291360 TI - Determinants of clinical response and survival in patients with congestive heart failure treated with captopril. AB - The efficacy of chronic ambulatory captopril (CPT) therapy was evaluated over an 18-month period in 36 patients with refractory chronic congestive heart failure (CHF) by cardiac catheterization, treadmill exercise, nuclear scintigraphy, echocardiography, and symptomatology. Clinical improvement to New York Heart Association functional class I or class II was observed in 63% of the patients (20 of 32) after 2 months of treatment; this amelioration of CHF symptoms was sustained in 63% of the patients (10 of 16) at 18 months. Exercise tolerance increased in 64% of the patients (16 of 25) at early follow-up and in 79% (11 of 14) at late follow-up. Univariate analysis revealed that the pre- and post-CPT stroke work indices (SWI) and the post-CPT cardiac index related to favorable long-term clinical response. Fourteen CHF patients (39%) died during the 18-month follow-up. Univariate analysis revealed that the pretreatment SWI, right atrial pressure, plasma norepinephrine concentration, and echocardiographic shortening fraction were significant predictors of mortality. Multivariate analysis indicated that the SWI was the principal determinant of survival: the 18-month cumulative survival rate for CHF patients with a SWI less than 32 gm . m/m2 was 44% compared to 88% when the SWI was greater than 32 gm . m/m2. Thus, CPT results in sustained symptomatic and functional improvements in patients with advanced CHF, but the mortality remains high and is primarily related to the severity of cardiac dysfunction. PMID- 6291361 TI - Hemodynamic effects of captropril in chronic heart failure: efficacy of low-dose treatment and comparison with prazosin. AB - The acute hemodynamic effects and long-term therapeutic actions of low doses of the oral angiotensin-converting enzyme (ACE) inhibitor captopril (CPT) were evaluated in 18 patients with severe chronic congestive heart failure (CHF). Increasing doses of 1, 2.5, 6.25, 12.5, and 25 mg of CPT were given at 2-hour intervals. Increased stroke volume index (SVI) and reduced mean pulmonary capillary wedge (PCW) pressure occurred at 1 hour (p less than 0.05) with an associated decline of blood pressure. Maximal hemodynamic improvement for the group was seen at 6 and 7 hours following the 6.25 and 12.5 mg doses when SVI was elevated 35% and mean PCW pressure decreased 40% from control. CPT in doses of 12.5 to 50 mg every 8 hours was continued long term in these 18 CHF patients. Four patients died, and one was noncompliant; drug therapy was withdrawn in two patients with symptomatic hypertension and in one patient who experienced an alteration in taste. The remaining 10 patients showed significant improvement in symptoms and treadmill exercise duration at 3 months after CPT therapy was started. Moreover, repeat hemodynamic measurements were similar to optimal measurements obtained during the initial study. In a further study, the acute hemodynamic and hormonal effect of sequentially randomized 5 mg prazosin and 25 mg CPT were compared in 10 CHF patients. While both drugs reduced PCW pressure and vascular resistance, CPT effects were greater. Further, CPT alone effected a small rise in cardiac index and minor decline in heart rate, and CPT diminished aldosterone levels and increased plasma renin activity while prazosin did not. PMID- 6291362 TI - Acute and long-term effects of captopril on exercise cardiac performance and exercise capacity in congestive heart failure. AB - Although many studies have shown that captopril (CPT) provides acute hemodynamic improvement in patients with severe congestive heart failure (CHF) at rest, little information is available concerning exercise hemodynamic responses to CPT or the effect of this drug on exercise tolerance in CHF. Therefore, we evaluated the hemodynamic effects of CPT at rest and during upright bicycle exercise in 15 patients with stable CHF. CPT (25 to 50 mg) reduces both resting heart rate and mean arterial pressure (84 +/- 11 to 78 +/- 7 bpm, p less than 0.025 and 85 +/- 9 to 64 +/- mm Hg, p less than 0.001). Concomitantly, left ventricular filling pressure dropped dramatically (26 +/- 9 to 15 +/- 7 mm Hg, p less than 0.001), while cardiac and stroke indices rose (2.0 +/- 0.5 to 2.5 +/- 0.6 L/min/m2, p less than 0.001, and 25 +/- 8 to 33 +/- 7 ml/m2, p less than 0.001). Similar directional changes occurred during exercise, with heart rate, mean arterial pressure, and left ventricular filling pressure at maximum exercise being less (123 +/- 15 to 115 +/- 16 bpm, p less than 0.01; 93 +/- 17 to 86 +/- 14 mm Hg, p less than 0.05; and 35 +/- 10 to 30 11 mm Hg, p less than 0.001, respectively) after CPT ingestion. Peak exercise cardiac index rose slightly (3.6 +/- 0.7 to 3.9 +/- 0.6 L/min/m2) but not significantly. Six patients followed long term on CPT underwent elective recatheterization after 3 months. In these, the beneficial hemodynamic changes seen acutely persisted or further improvement was noted, both at rest and during exercise. Most impressively, peak exercise cardiac index rose from 3.6 +/- 0.7 to 4.6 +/- 1.0 L/min/m2 (p less than 0.05), and this was associated with an increase in exercise duration (8.0 +/- 2.2 to 11.5 +/- 1.4 minutes, p less than 0.05) and exercise work load (332 +/- 32 to 468 +/- 52 kp m/min, p less than 0.05). These findings indicate that in patients with severe CHF, oral CPT provides markedly beneficial augmentation of cardiac function during activity as well as at rest; moreover, chronic CPT therapy substantially increases exercise capacity in this setting. PMID- 6291363 TI - Captopril pharmacokinetics and the acute hemodynamic and hormonal response in patients with severe chronic congestive heart failure. AB - Twelve patients with severe chronic congestive heart failure (CHF) underwent simultaneous evaluation of the pharmacokinetic, pharmacodynamic, and neurohumoral actions of a single 25 mg oral dose of the angiotensin-converting enzyme (ACE) inhibitor captopril (CPT). Following drug administration, which raised plasma renin activity (PRA) and thereby indicated significant ACE inhibition, both free (unchanged) and total CPT (including active metabolites) were detectable in the blood with 40 minutes and peak blood levels of the agent were recorded 1 hour after CPT. Total CPT concentration was higher and persisted longer than free CPT, which became virtually nondetectable 8 hours after ingestion. Concomitantly, left ventricular function was markedly augmented by the oral ACE inhibition in all patients, with the magnitude of this improvement being closely related to the baseline PRA. Thus, the overall hemodynamic response to CPT, which rapidly appears in the bloodstream following drug intake in patients with advanced CHF, is a function of the extent of baseline renin-angiotensin-aldosterone activity. PMID- 6291364 TI - Use of captopril to estimate renin-angiotensin-aldosterone activity in the pathophysiology of chronic heart failure. AB - Although the renin-angiotensin-aldosterone system has been implicated in the pathophysiology of chronic heart failure, the factors influencing the extent of renin activity have not been clarified. In the present study, we evaluated 34 patients with severe chronic congestive heart failure in terms of baseline plasma renin activity (PRA). No uniform relationships between PRA and urinary sodium excretion have been established. Although baseline PRA did not correlate with the baseline hemodynamic values, the extent of hemodynamic improvement following acute oral captopril (CPT) therapy (25 mg) was highly correlated with baseline PRA. The onset of action was rapid, so that the initial acute response could be readily assessed and at the same time could be used as a means to estimate baseline renin-angiotension-aldosterone activity. For long-term therapy, however, the response to CPT must be assessed in the context of several potentially simultaneous factors that might either enhance or compromise its effectiveness. PMID- 6291365 TI - The effect of captopril on postural hemodynamics and autonomic responses in chronic heart failure. AB - Both postural abnormalities and autonomic dysfunction have been identified in patients with chronic congestive heart failure (CHF). However, the effect of long term vasodilator therapy on these phenomena has not been assessed. In this study the hemodynamic and plasma norepinephrine (PNE) responses to upright posture, as well as the cold pressor test and Valsalva's maneuver in 12 patients with severe chronic CHF during both acute and long-term captopril (CPT) therapy, were evaluated. This revealed an absence of the normal hemodynamic adjustments to upright posture and a blunted response of PNE. The heart rate and blood pressure responses to the cold pressor test and Valsalva's maneuver were similarly blunted. The reflex adjustments of systemic resistance during tilt improved with CPT therapy, but the absence of reflex tachycardia in the upright posture persisted. Additionally, there was improvement of the PNE response, and the responses of heart rate and blood pressure to the cold pressor test were virtually normalized during long-term CPT therapy. The abnormal response to the Valsalva maneuver persisted. In conclusion, hemodynamic and reflex-mediated responses to upright posture and the standard assessment of autonomic control mechanisms revealed abnormal patterns in heart failure. While the hemodynamic adjustment to postural changes and sympathetic responsiveness were improved with CPT, complete correction of these abnormalities did not occur. Whether the improvement was a nonspecific vasodilator effect or the result of specific CPT therapy remains to be determined. PMID- 6291367 TI - The herpesviruses. PMID- 6291366 TI - Acute and long-term effects of captopril on left and right ventricular volumes and function in chronic heart failure. AB - To more fully delineate the effects of captopril (CPT) on cardiac size and function in patients with chronic congestive heart failure (CHF), we performed blood pool scintigraphy in conjunction with hemodynamic measurements during the initiation of vasodilator therapy and repeat scintigraphy after 2 and 12 weeks. Measurements of both right and left ventricular volumes and ejection fraction were made before and 90 minutes after the first 25 mg dose of CPT in 14 patients. Overall, heart rate fell modestly, from 82 +/- 10 to 77 +/- 11 bpm (p less than 0.01), while mean arterial pressure dropped considerably, from 86 +/- 10 to 73 +/ 13 mm Hg (p less than 0.01). Left ventricular end-diastolic and end-systolic volumes fell from 388 +/- 81 to 350 +/- 77 and 319 +/- 75 to 271 +/- 75 ml (both p less than 0.01). Right ventricular end-diastolic and end-systolic volumes also decreased. The left ventricular ejection fraction rose modestly but significantly, from 19 +/- 6% to 22 +2- 5% (p less than 0.01), as did right ventricular ejection fraction (25 +/- 9% to 29 +/- 11%, p less than 0.01). Simultaneous hemodynamic measurements demonstrated corresponding decreases in left and right ventricular filling pressures (24 +/- 10 to 17 +/- 9 to 10 +/- 5 to 6 +/- 5 mm Hg, respectively, both p less than 0.001). The slight increase in cardiac and stroke indices at 90 minutes, from 2.0 +/- 0.5 to 2.1 +/- 0.4 and 25 +/- 8 to 27 +/- 7 ml/m2, respectively, were not significant, although both indices rose significantly at the time of each individual's peak effect. Six patients were discharged on CPT and underwent follow-up nuclear studies: the acute decrease in ventricular volumes were sustained, as was the slight improvement in ejection fractions. These findings indicated that the major acute effect of CPT in CHF is a reduction in the preload of both ventricles. This reduction in diastolic volumes and pressures persists during chronic therapy. PMID- 6291369 TI - Scintigraphic prediction of pulmonary arterial systolic pressure by regional right ventricular ejection fraction during the second half of systole. AB - In 49 patients in whom gated equilibrium ventriculography and cardiac catheterization were performed within a 6 day interval, total and fractional portions of global and regional right ventricular ejection fraction (RVEF) were correlated with pulmonary arterial systolic pressure. Pulmonary arterial systolic pressure was normal (30 mm Hg or less) in 27 patients (Group I) and elevated (31 mm Hg or greater) in 22 patients (Group II). The second-half regional RVEF was 38 +/- 8% (mean +/- standard deviation) with a range of 30 to 54% for Group I and 22 +/- 6% with a range of 13 to 32% for Group II. The difference between the means was statistically significant (p less than 0.001). Use of a second-half regional RVEF of 30% as the criterion of elevated pulmonary arterial systolic pressure resulted in a sensitivity of 0.86 and a specificity of 1.00. A power curve fit in which pulmonary arterial systolic pressure = 10.91 (second-half regional RVEF) 0.87 allowed accurate estimation (r = -0.85) of pulmonary arterial systolic pressure from the second-half regional RVEF. It is concluded that second-half regional RVEF may be used to accurately detect pulmonary arterial hypertension and to estimate its extent. PMID- 6291368 TI - Amiodarone for control of sustained ventricular tachyarrhythmia: clinical and electrophysiologic effects in 51 patients. AB - We evaluated the electrophysiologic effects of amiodarone and its ability to control ventricular arrhythmia in a selected group of 51 patients with refractory sustained ventricular arrhythmia. Amiodarone in doses of 400 to 800 mg/day prolonged refractoriness in the atria, atrioventricular (AV) node, and ventricle as well as conduction through the AV node and His-Purkinje system. Although it had no effect on measurements of sinus nodal function (sinus nodal recovery time and sinoatrial conduction time), it prolonged the sinus cycle length and 2 patients required a permanent pacemaker for symptomatic sinus bradycardia. Amiodarone did not alter the ease of inducibility in any consistent manner, and only 5 of 43 patients (12%) who had inducible ventricular tachycardia before amiodarone therapy had none induced during amiodarone treatment. The clinical effectiveness of amiodarone could be evaluated in 46 patients followed up for 8.6 +/- 6 months (range 0.5 to 22). It provided effective therapy in 23 patients (50%), partly effective therapy in 13 (28%), and was ineffective in 10 (22%). Adverse effects were noted in 28 of 51 patients (55%), and in 11 of these (22%) the drug had to be discontinued because of adverse effects. We conclude that amiodarone is a useful agent for the treatment of refractory sustained ventricular arrhythmia. Its use should be reserved for patients with life threatening sustained arrhythmia because of the significant incidence of adverse effects. Furthermore, good clinical response can be observed in patients receiving amiodarone in spite of continued inducibility. PMID- 6291371 TI - Changes in work tolerance associated with metabolic and physiological adjustment to moderate and severe iron deficiency anemia. AB - The time course of metabolic and physiological adjustment to moderate iron deficiency anemia (MIDA, 8 g Hb/dl) and to severe iron deficiency anemia (SIDA, 4 g Hb/dl) was studied in adult, male Sprague-Dawley rats at 3, 7, 14, 30, 60, 90, 150, and 360 days, respectively. Our previous studies using the same rats used in the present study indicated that bone marrow iron was absent and plasma iron was significantly lower (p less than 0.001) in MIDA and SIDA relative to control. The following results with MIDA and SIDA rats are all expressed relative to control values. Red cell 2,3-diphosphoglycerate ranged from 45 to 146% greater in MIDA over the 360-day period and was 130% greater in SIDA at 30 days. Exhaustive run time consistently averaged 64 +/- 3% (SEM) less in MIDA over the time course and was further lowered to 18% in SIDA at 30 days. Heart rates of MIDA were elevated (p less than 0.05) at 180 days but lower (p less than 0.001) at 360 days in response to exercise. Resting heart rates of MIDA were the same at 180 and 360 days. Heart rates of SIDA were elevated (p less than 0.05) at rest and during exercise at 30 days. Organ weight/body weight changes indicated cardiomegaly in MIDA from 90 to 150 days which reverted to normal at 360 days; splenomegaly in MIDA from 90 to 360 days; and kidney atrophy in MIDA at 60 and 90 days which reverted to normal thereafter; in SIDA cardiomegaly and splenomegaly were present at 30 days. These results indicate that the onset and magnitude of physiological and metabolic adjustments occur in proportion to the severity of the anemia, and despite compensatory adjustments in parameters related to work performance, a new stable, lowered level of work tolerance is reached. PMID- 6291370 TI - Assessment of vasodilator therapy in patients with severe congestive heart failure: limitations of measurements of left ventricular ejection fraction and volumes. AB - Although noninvasive techniques are often used to assess the effect of vasodilator therapy in patients with congestive heart failure, it is unknown whether changes in noninvasively determined left ventricular ejection fraction, volume, or dimension reliably reflect alterations in intracardiac pressure and flow. Accordingly, we compared the acute effect of sodium nitroprusside on left ventricular volume and ejection fraction (determined scintigraphically) with its effect on intracardiac pressure and forward cardiac index (determined by thermodilution) in 12 patients with severe, chronic congestive heart failure and a markedly dilated left ventricle. Nitroprusside (infused at 1.3 +/- 1.1 [mean +/ standard deviation] microgram/kg/min) caused a decrease in mean systemic arterial, mean pulmonary arterial, and mean pulmonary capillary wedge pressure as well as a concomitant increase in forward cardiac index. Simultaneously, left ventricular end-diastolic and end-systolic volume indexes decreased, but the scintigraphically determined cardiac index did not change significantly. Left ventricular ejection fraction averaged 0.19 +/- 0.05 before nitroprusside administration and increased by less than 0.05 units in response to nitroprusside in 11 of 12 patients. The only significant correlation between scintigraphically and invasively determined variables was that between the percent change in end diastolic volume index and the percent change in pulmonary capillary wedge pressure (r = 0.68, p = 0.01). Although nitroprusside produced changes in scintigraphically determined left ventricular ejection fraction, end-systolic volume index, and cardiac index, these alterations bore no predictable relation to changes in intracardiac pressure, forward cardiac index, or vascular resistance. Furthermore, nitroprusside produced a considerably greater percent change in the invasively measured variables than in the scintigraphically determined ones. PMID- 6291372 TI - Vegetarianism, dietary fiber, and mortality. AB - A prospective study was set up to test the hypotheses that the risk of death from various diseases is reduced by a high intake of dietary fiber or by vegetarianism. A simple screening questionnaire was distributed among persons with a special interest in health foods, and 10,943 subjects were recruited and followed-up. Their mortality was ascertained by flagging their National Health Service records, and analyzed after 7 yr. A significant negative association was found between vegetarianism and mortality from ischemic heart disease which was especially marked among the men and did not seem to be due to a confounding effect of smoking. No significant associations were found with fiber, although persons who habitually ate wholemeal bread had a lower mortality from cerebrovascular disease. These findings confirm other evidence of a lower mortality from heart disease among vegetarians. PMID- 6291373 TI - Dietary determinants of lipoproteins, total cholesterol, viscosity, fibrinogen, and blood pressure. AB - High-density lipoprotein cholesterol, low-density lipoprotein cholesterol, total cholesterol, viscosity, fibrinogen, and blood pressure were determined in 117 men aged 44 to 60 yr selected from the general population who also completed 7-day weighed dietary records. Associations between these measurements and a number of dietary factors were assessed by multiple regression analysis, allowing where necessary for the effects of age, body mass index, and smoking habit. High density lipoprotein cholesterol was associated positively with both alcohol and fish consumption and negatively with saturated fat intake. High-density lipoprotein cholesterol expressed as a percentage of total cholesterol was associated negatively with the percentage of energy from fat and positively with fish consumption. Low-density lipoprotein cholesterol was associated positively with the percentage of energy from fat and negatively with fish consumption. Fibrinogen and systolic blood pressure were inversely related to cereal fiber intake. PMID- 6291374 TI - Evaluation of a self-administered dietary questionnaire for use in a cohort study. AB - A self-administered diet questionnaire designed for use in a cohort study has been assessed in comparison with a detailed quantitative diet history previously used in and validated for case-control studies. One hundred fifty-eight women aged 40 to 59 were asked to complete the self-administered questionnaire and 123 returned it by mail. Of these women, 50 were interviewed at home using the detailed diet history. Estimates of intake of major nutrients other than fat and fatty acids are comparable from the two methods although in general, estimated intake from the self-administered questionnaire was higher than the diet history. The difference in intake of fat and fatty acids was largely but not completely accounted for by differences in amounts of added fat recorded. It is concluded that when applied to large numbers of women in a cohort study, the self administered questionnaire is feasible to administer and will be sufficiently accurate in estimating nutrient intake as compared to estimates from a detailed, interview-type diet history. PMID- 6291375 TI - Automated cytochemical staining and inflammation. Further assessment of the "left shift". AB - Automated differential leukocyte counting devices identify neutrophils on the basis of size and peroxidase staining. Since no analysis of neutrophil subpopulations is provided, detection of the "left shift" as reflected by increased numbers of band neutrophils is not possible. It has been suggested that samples with increased numbers of high peroxidase neutrophils (HPX) correspond to conventional samples with increased band neutrophils. Using blood samples enriched by 700% in their band neutrophil concentration, no significant change in the number of HPX cells or mean peroxidase content was detected. Examination of Wright-stained smears with increased concentrations of HPX cells invariably revealed toxic neutrophils not seen in controls. The authors concluded that increases in HPX cells do reflect inflammation but that such increases correlate with the presence of toxic neutrophils and not band neutrophils. PMID- 6291376 TI - Adaptation of an automated microbiology system for the growth of anaerobes and performance of antimicrobial susceptibility tests. AB - The authors examined the feasibility of growing anaerobes in an automated microbiology system (MS-2) and using the system for antimicrobial susceptibility testing. A total of 78 of 100 clinical anaerobic isolates grew in the MS-2 if 100 microL of an undiluted overnight suspension of organisms was inoculated into at least 2.5 mL of freshly prepared Wilkins-Chalgren broth supplemented with 0.07% agar. Susceptibility test results were determined with 50 anaerobes tested against seven antibiotics in the MS-2 system and were compared with results determined with a qualitative reference susceptibility test method. of the 350 tests, 88% of the MS-2 results agreed with the qualitative reference results. False-susceptible and false-resistant results were reported for 8.6 and 3.4%, respectively, of the MS-2 results. PMID- 6291377 TI - Needle-shaped inclusions in plasma cells in a patient with hypogammaglobulinemia. AB - A case of a patient with needle-shaped inclusions in plasma cells was reported. Some of the inclusions were positively stained for acid phosphatase and beta glucuronidase. Ultrastructurally, each inclusion was surrounded by a single limiting membrane without any relation to rough-surfaced endoplasmic reticulum and composed of numerous fine fibrous bundles. By the enzyme-labeled antibody technic, the inclusions were found as "stain defects" in IgG-forming plasma cells, but not in IgM-forming plasma cells. Although the exact nature of the inclusions could not be clarified, they were quite different from both amyloid fibrils and immunoglobulin-derived inclusions, and were thought to be synthesized by a clone of differentiated plasma cells. The patient showed moderate hypogammaglobulinemia but no evidence of a direct correlation between the inclusions and hypogammaglobulinemia was obtained. Though the possibility that the case was non-secretory myeloma could not be ruled out, it was unlikely judged by the findings of various examinations. PMID- 6291378 TI - Endometriosis of the colon with mixed "germ cell" tumor. AB - A 42-year-old woman was found to have endometriosis of the oviducts. Five years later she had a hysterectomy and oophorectomy, the abnormal findings at this time being endometriosis with a chocolate cyst of one ovary. At a third operation, seven years after the first, she was found to have endometriosis of the colon, associated with malignant tumor in the serosa of the intestine and in the pelvis. Histologically, there was direct transition from benign epithelium in the ectopic endometrial glands, to dysplastic and neoplastic tissue. In different areas, the tumor included elements of yolk sac type (endodermal sinus tumor), choriocarcinoma, and teratoma. The implications of this unique transformation and combination are discussed briefly. PMID- 6291379 TI - Spontaneous rupture of the spleen secondary to metastatic hepatocellular carcinoma: a report of a case and review of the literature. AB - An extremely rare case of splenic rupture at the site of metastasis of hepatocellular carcinoma is reported. A 62-yr-old woman with hepatocellular carcinoma and its suspected metastasis to the spleen died of intraperitoneal hemorrhage. Autopsy disclosed a laceration of the spleen as the definitive cause of the hemorrhage. There were multiple nodules of metastatic hepatocellular carcinoma in the spleen, some of which were exposed at the lacerated portion of the splenic capsule. This may be the first report of a case of spontaneous rupture of the spleen secondary to metastatic hepatocellular carcinoma. Splenic rupture can be one of the causes of hemoperitoneum in patients with hepatocellular carcinoma. PMID- 6291380 TI - Change of enzyme properties caused by cross-linking treatment of human erythrocytes. AB - Aragon et al [4] reported that rat erythrocytes can be cross-linked and permeabilized without significant inactivation or alterations of several enzymes in the glycolytic pathway. If this is the case, in situ kinetic analysis of the red cell enzymes in normal human red cells and abnormal red cells associated with hemolytic problems could be performed. However, we found that the treatment of human red cells with several bifunctional cross-linking reagents under various conditions always caused a certain extent of inactivation and a change in kinetic properties of glucose-6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase, and lactate dehydrogenase. Thus, the cross-linking and permeabilization method, as it stands, is not satisfactory for in situ kinetic analysis of red cell enzymes. PMID- 6291381 TI - Second malignancies in hairy cell leukemia (leukemic reticuloendotheliosis). PMID- 6291382 TI - Tumours of the mononuclear phagocyte system: a review of clinical and pathological features. AB - Cells of the mononuclear-phagocyte system (MPS) may give rise to a wide variety of tumours of diverse morphology ranging from acute leukemia to small benign tumours of the skin. This review of tumours of the MPS was prompted by recent advances in understanding of tumour morphology and immunologic features. It is now possible to differentiate tumours of the MPS mor precisely from tumours of lymphoid origin with which they have often been confused; however, controversy still exists as to the origin of the malignant cell of Hodgkin disease. Current evidence is reviewed which favours the view tht the malignant cell of Hodgkin disease is of monocyte origin. A classification of MPS tumours based on organ system of origin and disease extent is presented. PMID- 6291383 TI - The role of macrophages and immunocytes in the pathogenesis of pulmonary diseases due to mineral dusts. AB - There is considerable evidence of generalized stimulation of the immune system in pulmonary dust diseases. This stimulation involves both the T- and the B-arms of the immune system. A reasonable explanation of this immune stimulation resides in an effect of the mineral dusts on the macrophages. This effect is likely to be mediated by the production of interleukin-1. Since the same substance also stimulates fibroblasts, a unified view of the pathogenesis of pulmonary dust diseases is now possible. PMID- 6291384 TI - Lung cancer and smoking in a group of iron ore miners. AB - Several studies have shown that miners, in both uranium and nonuranium mines, have an increased lung cancer mortality, probably caused by exposure to radon and its daughters. The excess mortality has been observed primarily among smoking miners but some recent studies have also indicated a considerably increased risk among nonsmoking miners. This study, among a group of iron ore miners, was undertaken to further elucidate the somewhat unclear and presumably complex relationship of mining, smoking, and lung cancer. The results show a 16-fold increase in lung cancer mortality among miners versus nonminers. Even nonsmoking miners seem to be at a rather high risk of developing lung cancer, but there was a tendency for the most heavy smoking miners to die earlier and to have a slightly shorter induction-latency period for development of lung cancer than was found among the nonsmoking miners. PMID- 6291385 TI - Accelerated loss of FEV- in polyurethane production workers: a four-year prospective study. AB - A four-year longitudinal study of ventilatory function in polyurethane-foam production workers exposed to toluene diisocyanate (TDI) revealed a dose-response relationship between average exposure to TDI and change in forced expiratory volume per second (FEV-1). Workers with mean exposure in excess of 0.0035 ppm showed a greater rate of decline of FEV-1 over the four-year period than that expected from aging. Factors other than TDI exposure (sex, smoking history, history of atopy) do not account for the loss. The current threshold limit value (TLV) for exposure to TDI in industry (0.02 ppm) does not protect workers from accelerated impairment of ventilatory capacity. PMID- 6291386 TI - Use of conventional and IgM-specific radioimmunoassays for anti-hepatitis A antibody in an outbreak of hepatitis A. AB - During a common source outbreak of hepatitis A, we studied the characteristics and utility of commercially available radioimmunoassays for total and IgM specific antibody to hepatitis A virus. IgM hepatitis A antibody was detectable in all serum specimens obtained up to 119 days following onset from the seven persons with hepatitis A, and as long as 347 days in one person. Acute infection could also be documented by a four-fold or greater increase in titers of hepatitis A antibody, although as long as nine weeks was required between the times acute and convalescent specimens were obtained. The radioimmunoassay for IgM-specific hepatitis A antibody had greater specificity (99 percent versus 84 percent) and a higher positive prediction value (88 percent versus 23 percent) for the diagnosis of acute hepatitis A than did the radioimmunoassy for hepatitis A antibody. Uses of the radioimmunoassay for IgM-specific hepatitis A antibody include rapid diagnosis of acute hepatitis A, differentiation between recent and past hepatitis A infection, and screening for recent hepatitis A infection in epidemiologic investigation. PMID- 6291387 TI - Specific cell-mediated immunity and infections with herpes viruses in cardiac transplant recipients. AB - Immune responses and infections with herpes viruses were studied prospectively in 36 cardiac transplant recipients. Specific lymphocyte transformation and interferon production in response to viral antigens, viral culture results, antibody levels, responses to phytohemagglutinin, and T-cell numbers were determined. Responses to phytohemagglutinin and T-cell numbers were depressed for six to 12 weeks. Cytomegalovirus infection occurred in 100 percent of seropositive patients and in 62 percent of seronegative patients. Primary infection was more frequently symptomatic. Heart implantation from a seropositive patient wwas significantly correlated with subsequent infection in seronegative patients. Depression of transformation in response to cytomegalovirus correlated with prolonged shedding. Herpes simplex infection occurred in 95 percent of seropositive patients but decreased after 12 weeks. Asymptomatic shedding was rare, and primary infection did not occur. Return of transformation in response to herpes simplex was associated with decreased infection. Herpes zoster occurred in 22 percent during the first year, and transformation responses to varicella zoster returned thereafter. Depression of interferon production in response to viruses did not correlate with infection as well as did lymphocyte transformation. PMID- 6291388 TI - Hyperkalemia in azotemic patients during angiotensin-converting enzyme inhibition and aldosterone reduction with captopril. AB - Thirty-three hypertensive patients with a wide range of renal function were studied during initiation of angiotensin-converting enzyme inhibition with captopril to evaluate changes in potassium levels concomitant with reduction of aldosterone excretion. Ten patients (Group I) with low levels of plasma renin activity had no change in either aldosterone excretion or potassium during the first week of therapy. Twenty-three other patients (Group II) had decreased aldosterone excretion of an average of 63 percent, often reversing secondary hyperaldosteronism. This was associated with a rise in serum potassium from 3.6 +/- 0.1 to 4.4 +/- 0.1 mEq/liter (p less than 0.001). Serum potassium levels during captopril therapy were inversely related to glomerular filtration rate (creatinine clearance) and transiently exceeded 6.0 mEq/liter in markedly azotemic subjects. Despite rising potassium levels, nine patients had reduced aldosterone excretion to subnormal levels, sometimes for many months. During initiation of converting-enzyme inhibition, potassium-sparing agents and supplements should be discontinued and serum potassium levels should be monitored closely, particularly in patients with imparied renal function. PMID- 6291389 TI - Humoral hypercalcemia of malignancy: a syndrome in search of a hormone. AB - A women with hypercalcemia and a hypernephroma confined to the left kidney underwent nephrectomy and subsequent resolution of hypercalcemia. Serum parathyroid hormone was undetectable in peripheral blood as well as in the left renal vein at surgery. Parathyroid hormone was also undetectable in the tumor extract using three different antisera to parathyroid hormone. Measurement of plasma prostaglandin E and 13, 14-dihydro-15-keto-prostaglandin E2 revealed levels within the normal range. The serum 1,25-dihydroxyvitamin D concentration was below normal and nephrogenous cyclic adenosine monophosphate was markedly elevated. The humoral agent responsible for hypercalcemia in this patient was not identified. This case emphasizes the need to search for new hypercalcemic factors in patients with hypercalcemia of malignancy. PMID- 6291390 TI - Zoster encephalitis. Isolation of virus and measurement of varicella-zoster specific antibodies in cerebrospinal fluid. AB - Varicella-zoster virus (VZV) was isolated on two occasions from the cerebrospinal fluid of an elderly woman with encephalomyelitis complicating thoracic zoster. Antibodies to ZV-induced membrane antigen (FAMA) were present in cerebrospinal fluid in a titer of 1:64; serum antibodies were 64-fold higher. Further evidence for local antibody production was derived from simultaneous measurements of immunoglobulin G and albumin in cerebrospinal fluid and serum and calculation of a cerebrospinal fluid-IgG index. PMID- 6291391 TI - Comparison of pharmacodynamic properties of various estrogen formulations. AB - A group of 23 healthy postmenopausal women received one or more 2-week courses of daily administration of the following estrogen preparations: piperazine estrone sulfate (Ogen), 0.3, 0.625, 1.25, 2.5, and 5.0 mg; micronized estradiol (Estrace), 1, 2, and 10 mg; conjugated estrogens (Premarin), 0.3, 0.625, 1.25, and 2.5 mg; ethinyl estradiol (Estinyl), 10 and 20 micrograms; and diethylstilbestrol, 0.1 and 0.5 mg. Each dosage of each formulation was ingested by three women. In those women who received more than one dosage, each course was separated by a drug-free interval of at least 4 weeks. Pretreatment and posttreatment levels of follicle-stimulating hormone (FSH), luteinizing hormone (LH), corticosteroid-binding globulin-binding capacity, sex hormone-binding globulin-binding capacity, angiotensinogen, estrone, and estradiol were determined. The relative potency of these five estrogen formulations was determined by parallel line analysis for each of these responses, except LH. On a weight basis, piperazine estrone sulfate and micronized estradiol were equipotent for all responses. Conjugated estrogens suppressed FSH in a fashion equipotent to that of the other nonsynthetic estrogens; however, for all three hepatic parameters, the response was exaggerated twofold to threefold. The synthetic estrogens, diethylstilbestrol and ethinyl estradiol, were relatively more potent on a weight basis for every response and produced the most marked response (fourfold to eighteenfold in excess of their FSH suppression) for the hepatic parameters. PMID- 6291392 TI - Amniotic fluid beta-endorphin and alpha-melanocyte--stimulating hormone immunoreactivity in normal and complicated pregnancies. AB - beta-Endorphin (beta-EP) and alpha-melanocyte--stimulating hormone (alpha-MSH) are members of a family of peptides which are found in the intermediate lobe of the fetal pituitary gland and placenta. In the present study, concentrations of immunoreactive beta-EP (i beta-EP) and alpha-MSH were determined by radioimmunoassay in the amniotic fluid compartment of both normal (n = 72) and complicated (n = 44) pregnancies. In normal pregnancies, there was a significant (p less than 0.05) fall in the mean amniotic fluid i beta-EP from the second to third trimester (from 175 to 132 pg/ml). A similar, but not statistically significant, decline in the mean amniotic fluid concentrations of alpha-MSH (from 46 to 34 pg/ml) was also noted. Concentrations of amniotic fluid i beta-EP showed a significant correlation with alpha-MSH (r = 0.61, p less than 0.001). Significant elevations of mean amniotic fluid i beta-EP but not alpha-MSH were found in pregnancies complicated by intrauterine growth retardation (215 pg/ml, p less than 0.001) and premature labor (225 pg/ml, p less than 0.001). Our findings suggest that levels of i beta-EP and alpha-MSH in amniotic fluid decrease from the second to the third trimester of pregnancy, and that elevation in i beta-EP concentration in amniotic fluid, in all probability, reflects the secretion of beta-EP by the fetus in response to fetal distress. PMID- 6291393 TI - beta-Endorphin and beta-lipotropin concentrations in umbilical cord blood. AB - Antisera suitable for human beta-endorphin and beta-lipotropin radioimmunoassay were developed, and radioimmunoassays were established to measure these peptides in umbilical cord plasma, with silicic acid extraction and gel chromatography used to separate the beta-endorphin from the beta-lipotropin fraction. These two peptides were determined in umbilical venous plasma from 64 newborn infants. Umbilical vein beta-endorphin and beta-lipotropin concentrations averaged 38.5 +/ 3.2 and 50.4 +/- 4.1 (+/- SE) fmoles/ml in the 54 newborn infants without and 115 +/- 18 and 110 +/- 25 fmoles/ml in the 10 newborn infants with apparent fetal distress. Neither the presence or absence of labor nor the route or mode of delivery was found to affect umbilical vein beta-endorphin or beta-lipotropin concentrations. However, cord plasma levels of both peptides were significantly elevated in conjunction with fetal distress, as evidenced by prolonged bradycardia, late and prolonged variable fetal heart rate decelerations, or fetal acidosis. In 18 of 22 pairs of simultaneously measured umbilical venous and arterial beta-endorphin and beta-lipotropin concentrations in newborn infants without apparent intrapartum distress, the venous beta-endorphin concentrations, which averaged 40.4 +/- 3.5 fmoles/ml, were significantly higher than the arterial beta-endorphin levels, with a mean of 28.5 +/- 4.2 fmoles/ml. No significant umbilical arteriovenous concentration difference could be observed for beta-lipotropin. This suggests that at least a portion of the coad plasma beta-endorphin is derived from the placenta. The ratio of umbilical arterial to venous beta-endorphin concentrations rose as the absolute cord plasma beta endorphin levels increased. Furthermore, both the molar umbilical venous and arterial beta-lipotropin to beta-endorphin ratios decreased significantly in association with intrapartum fetal distress. These data indicate tat the stress related increase in umbilical plasma beta-endorphin exceeds that of beta lipotropin and may be, at least in part, of fetal origin. Umbilical venous beta endorphin and beta-lipotropin levels of neonates whose mothers did not receive meperidine or other narcotics agents did not differ from those of neonates whose mothers were given meperidine or other narcotics during labor. Our data, in conjunction with those of others, are consistent with the hypothesis that fetal hypoxia causes the release of neurotransmitters such as beta-endorphin, which may modulate the regulation of fetal heart rate patterns. PMID- 6291394 TI - The effect of estrogen and progesterone on beta-adrenergic receptor activity in rabbit lung tissue. AB - Study of the effect of sex steroids on the development of beta-adrenergic receptors may be essential to an understanding of the mechanisms of both lung maturation and the initiation of labor. (3H)Dihydroalprenolol (DHA) was used to quantify beta-adrenergic receptor sites in mature and immature rabbit lung tissue. DHA binding was rapid, saturable, of high affinity (dissociation constant = 2 nM), and of low capacity (246 to 576 fmoles/mg of protein), and a adrenergic competitors demonstrated both stereoselectivity ([-]isomer much greater than [+]isomer) and a rank order of potency (isoproterenol much greater than norepinephrine greater than epinephrine) characteristic of the beta-adrenergic receptor. beta-Adrenergic receptors in the lung tissue of both mature and immature female New Zealand White rabbits were investigated under various sex steroid situations. Estrogen (diethylstilbestrol, 5 micrograms/day for 10 days) increased the beta-adrenergic receptor site number in immature rabbits compared to matched controls (435 versus 339 fmoles/mg of protein, p less than 0.02 by paired t test). Addition of progesterone (diethylstilbestrol, 5 micrograms/day for 10 days, plus progesterone, 5 mg/day for 3 days) returned the beta-adrenergic receptor site number to control values (321 fmoles/mg of protein, p less than 0.01). In mature rabbits, treatment with progesterone alone (10 mg/day for 4 days) caused a significant reduction in beta-adrenergic receptor site numbers compared to untreated, matched controls (357 versus 493 fmoles/mg of protein, p less than 0.05 by paired t test). in the presence of estrogen, beta-adrenergic receptor activity is enhanced in both mature and immature rabbit lung tissue. Addition of progesterone restores this activity to control values. PMID- 6291395 TI - Cortisporin dispensing error. PMID- 6291396 TI - Selected mutants of mouse hepatitis virus type 4 (JHM strain) induce different CNS diseases. Pathobiology of disease induced by wild type and mutants ts8 and ts15 in BALB/c and SJL/J mice. AB - The model system of central nervous system (CNS) disease induced by mouse hepatitis virus type 4 (MHV-4) is explored by comparison of wild type (wt) MHV-4 and two temperature-sensitive (ts) mutants, designated ts8 and ts15, in BALB/c and SJL/J mice. In BALB/c mice, 3 plaque-forming units (PFU) of wt MHV-4 given intracerebrally caused fatal encephalomyelitis in all mice by 7 days after infection, with spread of virus outside the CNS, especially to liver. In SJL/J mice, 3 PFU of wt virus was cleared within 2-3 days, with little spread, and up tp 100 PFU failed to cause fatal encephalomyelitis. However, larger amounts of virus, like 1000 PFU, caused fatal encephalomyelitis in SJL/J mice. In contrast, 10(4) PFU of MHV-4 ts8 did not cause death in either BALB/c or SJL/J mice, and persisted in the CNS of both strains while retaining its ts phenotype. There was significatnly less spread of virus outside the CNS. BALB/c mice usually showed demyelination, remyelination, and recurrent demyelination with ts8, while SJL/J mice only rarely had lesions. Intracerebral inoculation with 10(4) PFU of MHV-4 ts15 was associated with a persistent infection in CNS and liver of BALB/c mice; however, only occasional demyelination and hepatic lesions occurred. TS15 did not cause death in either BALB/c or SJL/J mice and did not cause histopathologic injury in SJL/J mice. PMID- 6291397 TI - Changes in Schwann cells and vessels in lead neuropathy. AB - Transmission electron microscopy (TEM) of peripheral nerve in rats receiving 6% lead carbonate for 4-10 weeks provided evidence of a specific Schwann cell injury, associated with demyelination. Intranuclear inclusions in Schwann cells appeared within 2 weeks of administration of a lead-containing diet. Swelling of Schwann cells and disintegration of their cytoplasm was evident at 4 weeks. Distinctive electron-dense inclusions appeared in both Schwann and endothelial cells during the period of intoxication and were ultrastructurally identical to pathognomonic inclusions of lead poisoning seen in renal tubular epithelial cells. Scanning microscopy (SEM) with electron-probe microanalysis was used to identify the lead-containing deposits. In addition to Schwann cell changes, vessels revealed endothelial cell injury and alteread permeability to macromolecules. Since morphologic changes of Schwann cells precede the development of altered vascular permeability and endoneurial edema, it appears that lead gains access to the endoneurium prior to the development of altered vascular permeability, suggesting that edema and altered endoneurial fluid pressure are epiphenomena that supervene after demyelination occurs. Remyelination, Schwann cell proliferation and formation of onion bulbs are manifestations of persistent toxic injury to myelin-sustaining cells, resulting in chronic demyelination. PMID- 6291399 TI - Regulation of prostaglandin biosynthesis in cultured cells. PMID- 6291398 TI - Metastasizing mammary tumors. Spontaneous mammary tumors in asplenic mice heterozygous for athymia. PMID- 6291400 TI - Regulation of polyamine content in cultured fibroblasts. AB - The content of putrescine and of the polyamines (spermidine and spermine) and the activities of their biosynthetic enzymes were measured in 3T3 mouse fibroblasts and SV40-transformed mouse fibroblasts over the entire period from subculturing in fresh medium until confluence. The transformed cells had a substantially higher content of putrescine and spermidine than the 3T3 cells and higher activities of all of the biosynthetic enzymes. However, the ratio of spermine synthase to spermidine synthase was higher in the 3T3 cells, which correlated with their higher spermine-to-spermidine ratio. All of the biosynthetic enzymes increased in activity during cell growth. Ornithine decarboxylase increased 20 fold with a maximum at 24-36 h after culturing whereas S-adenosylmethionine decarboxylase increased 3-fold at the same time. Spermidine synthase increased 10 to 16-fold during the growth period whereas spermine synthase increased 2- to 3 fold. The relative enzyme activities and the changes in total polyamine content suggested that 1) the activity of S-adenosylmethionine decarboxylase limited the production of the polyamines and 2) the relative amounts of spermidine and spermine synthase determined the predominant polyamine that the available decarboxylated S-adenosylmethionine is used to synthesize. When 3T3 cells become quiescent at confluence, there was a substantial fall in the intracellular spermidine level because of a greatly increased excretion of spermidine into the medium. Spermine content also fell because there was an increased conversion of spermine into spermidine, which was then excreted. The specific excretion of spermidine did not occur with the transformed SV-3T3 cells. PMID- 6291401 TI - Regulation of purine metabolism by plasma membrane and cytoplasmic 5' nucleotidases. AB - The contribution of plasma membrane 5'-nucleotidase (E.C. 3.1.3.5) to intracellular purine degradation and release was evaluated in cultured human lymphoblasts. B-lymphoblasts and T-lymphoblasts are characterized by high and low levels of plasma membrane 5'-nucleotidase activity, respectively. After radiolabeling of the cellular adenine nucleotide pools with [8-14C]adenine, deoxyglucose-induced purine nucleotide degradation resulted in a 2-2.5 times greater release of cellular radioactivity from the B-lymphoblasts than from the T lymphoblasts. Specific inhibition of plasma membrane 5'-nucleotidase with 50 microM alpha, beta-methylene adenosine diphosphate (AMPCP) did not decrease purine release during deoxyglucose-induced nucleotide degradation. Similarly, the inhibition of B-lymphoblast membrane 5-nucleotidase did not alter the incorporation of [8-14C]adenine into the nucleotide pool. Therefore, to explain the relatively high release of purine nucleotide degradation products in B lymphoblasts when compared with T-lymphoblasts, cytoplasmic 5'-nucleotidase activity was investigated in these cell lines. B-lymphoblasts have seven times more cytoplasmic 5'-nucleotidase activity for dAMP and two to three times more activity for other purine nucleoside 5'-monophosphates than do T-lymphoblasts at pH 7.4. Membrane and cytoplasmic nucleotidase activities are produced by different enzymes that can be distinguished by differences in pH optima, Michaelis constants for purine substrates, divalent cation requirements, and susceptibilities to AMPCP inhibition. The data suggest that plasma membrane 5' nucleotidase hydrolyzes extracellular nucleoside 5'-monophosphates only. Cytoplasmic 5'-nucleotidase most likely regulates the degradation of intracellular nucleoside 5'-monophosphates and may be responsible for the increased purine release observed in B-lymphoblasts. PMID- 6291402 TI - Enhanced receptor-cyclase coupling and augmented catecholamine-stimulated lipolysis in exercising rats. AB - To test the hypothesis that alterations of adipocyte beta-adrenergic receptors provide a molecular mechanism for enhanced catecholamine-stimulated lipolysis in physically trained animals, we studied adipocytes derived from rats subjected to 14 wk of swimming and from sedentary controls. Peak glycerol release and peak adenylate cyclase activity in response to epinephrine were increased in swimmers to 255% (P less than 0.01) and 156% (P less than 0.01) of control values, respectively, but neither basal glycerol release, basal cyclase activity, NaF stimulated cyclase activity, beta-receptor number, nor receptor affinity for [3H]dihydroalprenolol were altered. Epinephrine-stimulated adenylate cyclase activity remained increased in adipocytes from swimmers in the presence of theophylline or adenosine. In the absence of exogenous guanine nucleotide, we observed no differences in the dissociation constants for either the high affinity (KD = 0.025 microM) or the low-affinity (KL = 11 microM) classes of binding sites for (-)-epinephrine, but the proportion of high-affinity sites was greater in membrane preparations from swimmers than from controls (74 vs. 42%; P less than 0.01). We conclude that receptor-cyclase coupling is enhanced in adipocytes from exercising rats, perhaps due to an improved ability of adrenergic agonists to form the guanine nucleotide reversible high-affinity agonist-receptor complex. PMID- 6291403 TI - Effect of antimicrotubule agents on microtubules and steroidogenesis in luteal cells. AB - This report represents an effort to reinvestigate the relationship of steroid hormone processing with microtubule protein, using the highly active progesterone producing cells of superovulated immature rats as a model steroid hormone system. For the most part, gonadotropin-primed rats were used at 6 days and were reinjected with saline or various doses of colchicine and vinblastine: after 3 h the animals were injected with human chorionic gonadotropin (hCG, 10 IU) or saline for an additional hour. Subsequently, the ovaries were perfused with fixative for morphological studies or luteal cells were isolated for in vitro incubation with various additives. The results are as follows: 1) luteal cell progesterone synthesis is reduced in a dose- and time-dependent manner after treatment with antimicrotubule drugs; 2) additional stimulating agents given in vitro (hCG, Bt2cAMP, epinephrine) do not overcome the reduction produced by treatment with antimicrotubule drugs; 3) luteal cells show normal protein synthesis after treatment with antimicrotubule drugs; 4) luteal cells have microtubules of unusual appearance that are insensitive to the action of antimicrotubule agents. These results show that in the luteinized ovary, both colchicine and vinblastine interfere with hormone-stimulated steroidogenesis. Although the mechanism for the effect is not yet understood, it does not appear to be related to the content of the intact microtubules in luteal cells. Whether the drugs exert their action by binding to the unassembled form of microtubule protein in the cells remains to be determined. PMID- 6291404 TI - Physical training of lean and genetically obese Zucker rats: effect on fat cell metabolism. AB - The effects of 6-wk treadmill training program on the metabolism of isolated adipose cells from obese (fa/fa) and lean (Fa/?) Zucker rats were studied. Glucose metabolism and transport, insulin binding, and lipolysis were measured in adipose cells prepared from sedentary control and exercise-trained (ET) lean and/or obese rats. Two- to threefold increases in glucose metabolism were observed in cells from lean and obese ET rats compared with their respective controls. However, the insulin concentrations giving half-maximal stimulation (measuring insulin sensitivity) did not change (approximately 8 microunits/ml in lean and approximately 45 microunits/ml in obese rats). In lean ET rats, glucose transport and maximal glucose metabolic capacity (transport not rate-limiting) were increased twofold and sensitivity of lipolysis to epinephrine was increased three- to fourfold. These were not measured in obese rats. The results suggest that training of both lean and obese Zucker rats increases glucose utilization in adipose cells by increasing both glucose transport and intracellular glucose metabolism. Increased triglyceride turnover is also suggested by the increased sensitivity of lipolysis to epinephrine. PMID- 6291405 TI - Renal function and renal afferent and efferent nerve activity. AB - Recent microperfusion studies have fully substantiated the direct action of catecholamines on renal tubular reabsorptive rates. Surprisingly, these techniques have not provided consistent information on the nature of the adrenoceptor responsible for the stimulation of proximal tubular reabsorption. Both alpha- and beta-receptors have been favored for this role. These techniques have confirmed earlier reports that dopamine may have a direct natriuretic action on the renal tubules. The demonstration that renal efferent nerves contain both noradrenergic and dopaminergic fibers lends further support for the participation of dopamine in the regulation of salt and water excretion. Efferent renal nerve activity is modulated by a number of different afferent inputs to the central nervous system. One of these is the renal afferent innervation, which is composed of both chemoreceptor and mechanoreceptor fibers. A number of different reflexes that affect efferent renal nerve activity have been identified by electrical stimulation of renal afferent nerves or by selective stimulation of renal mechanoreceptors and chemoreceptors. These renorenal reflexes may have importance in the coordination of excretory activity between the two kidneys. Studies of these aspects of renal nerve function are reviewed. The importance of the renal nerves in conscious animals is also discussed in the light of evidence that their influence on renal function may be more apparent in abnormal or pathological circumstances. PMID- 6291406 TI - Glucocorticoid stimulation of Na-K-ATPase in superfused distal segments of kidney tubules in vitro. AB - The ability of glucocorticoids to regulate Na-K-ATPase activity directly was assessed in separated rat kidney tubules derived from the distal nephron. These tubules were superfused under sterile conditions and maintained in a viable condition for at least 24 h in a newly devised apparatus. Viability was assessed by measuring O2 consumption, protein/DNA ratios, and Na-K-ATPase and Mg-ATPase activities. At a concentration of 10(-8) M, dexamethasone elicited a 27% increase in tubular Na-K-ATPase activity in 6 h and a 32% increase in 24 h. In a separate series, assayed at 24 h, the maximal effect was obtained at a dexamethasone concentration of less than 10(-8) M, and by inspection half-maximal stimulation was obtained at approximately 10(-9) M. At a concentration of 10(-7) M, 17 beta estradiol, testosterone, progesterone, and deoxycorticosterone acetate had no significant effect on tubular Na-K-ATPase activity. These results as well as the time-course and dose-response data imply that the response is mediated by the glucocorticoid receptor pathway. Since the magnitude of response in vitro was similar to the one obtained after injection of dexamethasone in vivo, much if not all of the action appears to be direct and independent of glucocorticoid-induced changes in the filtered Na+ load. PMID- 6291407 TI - Effect of respiratory alkalosis on renal phosphate excretion. AB - Respiratory alkalosis induced hypophosphatemia and hypophosphaturia in intact animals. The present studies evaluated the effect of respiratory alkalosis on tissue phosphate distribution and renal phosphate transport in the presence and absence of parathyroid hormone (PTH). Respiratory alkalosis decreased plasma phosphate concentration and increased phosphate concentrations in muscle and liver. It decreased fractional phosphate excretion (FEPi) from 6.1 +/- 1.4 to 0.6 +/- 0.2%. In thyroparathyroidectomized (TPTX) rats infused with 20 mM phosphate, respiratory alkalosis decreased FEPi from 15.0 +/- 0.9 to 5.5 +/- 0.1%. PTH or dibutyryl cAMP administration produced a phosphaturia that was blunted by respiratory alkalosis. The phosphaturic response to PTH was also blunted in hypocapnic rats in which alkalosis was prevented by infusion of HCl. We conclude that respiratory alkalosis increases phosphate uptake by muscle, which largely accounts for the hypophosphatemia. The kidney response with increased phosphate reabsorption independent of plasma and kidney phosphate concentrations and with refractoriness to the phosphaturic effect of PTH. This refractoriness to the phosphaturic effect of PTH is due to decreased PCO2 rather than to the concomitant extracellular alkalosis. PMID- 6291408 TI - Effects of calcium on ADH action in the cortical collecting tubule perfused in vitro. AB - To test the effects of calcium on ADH action in an in vitro mammalian system, the rabbit cortical collecting tubule was studied. After 25 microunits/ml ADH (n=8) in the presence of 1.25 mM calcium bath, water flow (Jv) rose to 1.56 +/- 0.34 nl.mm-1. min-1 and hydraulic conductivity (Lp, cm.s-1.atm-1 X 10(7)) rose to 123 +/- 22. After 25 microunits/ml ADH in the presence of 3.75 mM calcium bath (n=7), Jv rose to 2.96 +/- 0.6 nl.mm-1.min-1 (P less than 0.05 vs. control) and Lp rose to 286 +/- 62 cm.s-1.atm-1 X 10(7) (P less than 0.02 vs. 1.25 mM bath calcium control). Tubules (n=6) perfused with 3.75 mM Ca and bathed in 3.75 mM Ca also showed an Lp of 279 +/- 82 cm.s-1.atm-1 X 10 (7) following 25 microunits/ml ADH. Tubules similarly studied in 1.25 (n=6) or 3.75 mM Ca (n=6) bath but treated with 10(-4) M 8-[p-chlorophenylthio]cAMP demonstrated Lp of 222 +/- 26 and 235 +/- 37 cm.s-1.atm-1 X 10(7), respectively. These data suggest that increased bath Ca enhances ADH- but not cAMP-stimulated water flow in the rabbit cortical collecting tubule. High perfusate Ca2+ does not alter the stimulatory effect of elevated peritubular Ca2+. PMID- 6291409 TI - Hydroosmotic response of collecting tubules to ADH or cAMP at reduced peritubular sodium. AB - Changes in cytosolic Ca2+ activity have been implicated in the hydroosmotic response to vasopressin (VP) in amphibian urinary bladder; the level of cytosolic free Ca2+ may be regulated, in part, by a process of Na-Ca exchange across the basolateral cell membrane. To assess whether similar mechanisms operate in the mammalian nephron, the effect of low peritubular [NA] on the hydraulic conductivity (Lp) of the isolated perfused rabbit collecting tubule (CT) exposed to either 20 microunits/ml VP or 5 X 10(-5) M 8-[p-chlorophenylthio]-cyclic 3',5' adenosine monophosphate (ClPheS-cAMP) was studied. Low peritubular [Na] had no effect on the basal water permeability of the CT. After exposure to VP, CTs bathed in 145 mM Na Ringer solution developed an Lp of 324 +/- 27 X 10(-7) cm.s 1.atm-1, while tubules bathed in 4 mM Na and 141 mM tetramethylammonium Ringer solution achieved an Lp of only 112 +/- 13 X 10(-7) cm.s-1.atm-1 (P less than 0.001). Inhibition of the VP response was estimated to be half-maximal when peritubular Na was 120 mM. The hydroosmotic response to ClPheS-cAMP was diminished by 44% of the control values when CTs were bathed in a 4 mM Na medium; this inhibition was greatly attenuated when the peritubular Ca concentration was reduced to 0.05 mM. These results are consistent with the view that 1) a Na-Ca exchange process operates at the basolateral surface of the mammalian cortical collecting tubule cells, and 2) elevated cytosolic Ca ion activity inhibits the increase in water permeability elicited by VP or cAMP in this nephron segment. PMID- 6291411 TI - Sympathoadrenal mechanisms in hemodynamic responses to gastric distension in cats. AB - There is presently little information on the efferent mechanisms responsible for the reflex cardiovascular activation during passive gastric distension. Therefore, 40 cats anesthetized with alpha-chloralose were studied with passive gastric balloon distention before and during 1) two repeated gastric distensions, 2) beta-adrenergic blockade with propranolol, 3) alpha-adrenergic blockade with phentolamine, or 4) bilateral adrenalectomy. Before and during each distension mean arterial pressure, heart rate, cardiac output, rate of rise of left ventricular pressure (dP/dt) at 40 mmHg developed pressure and calculated systemic vascular resistance were determined. Repeated gastric distension caused similar hemodynamic responses without tachyphylaxis. beta-Blockade significantly reduced the increase in dP/dt from 893 +/- 362 to 150 +/- 63 mmHg/s. alpha Blockade significantly altered the changes in mean arterial pressure from 33 +/- 5.0 to -2 +/- 4.7 mmHg and systemic vascular resistance from 0.114 +/- 0.019 to 0.004 +/- 0.031 peripheral resistance units. Bilateral adrenalectomy significantly diminished the contractile response from 525 +/- 107 to 50 +/- 85 mmHg/s but did not significantly alter the pressor and vasoconstrictor responses. We conclude that, during passive gastric distension in cats, the increase in myocardial contractility is mediated by beta-adrenergic-receptor stimulation, whereas the arterial vasoconstrictor and pressor responses are mediated by alpha adrenergic receptor stimulation. Additionally, during gastric distension a substantial portion of the contractile response is dependent on the integrity of the adrenal glands. PMID- 6291410 TI - Factors modifying contraction-relaxation cycle in vascular smooth muscles. AB - Contraction-relaxation cycles in vascular smooth muscles are largely dependent on the regulation of free Ca2+ in the myoplasm, as is the case in skeletal and cardiac muscles. In this article we describe the varieties of contraction relaxation cycles of vascular smooth muscles determined at cellular and subcellular levels. To discuss the excitation-contraction and pharmacomechanical coupling mechanisms in vascular tissues, passive and active membrane properties and ionic movements measured by various procedures are briefly introduced. In vascular smooth muscles the sources of Ca2+ contributing to the activation of contractile proteins are extra- and intracellular. Influxes of Ca2+ across the membrane are enhanced by the calcium spike and electrical and chemical depolarizations or activations of autonomic receptors.l However, the Ca2+ influx during the generation of action potential does not directly increase the free Ca2+ in the cell; rather, this ion is sequestered in the storage site and activates the calcium-induced calcium-release mechanism in the storage sites with a subsequent increase in the levels of free Ca2+. In some vascular tissues depolarizations induced by activations of autonomic receptors are not a prerequisite for generation of the contraction, as these mechanical responses appear with hyperpolarization of the membrane or without a change in the membrane potential. Possible functional links between the myoplasmic membrane where the receptors are distributed and the Ca2+ storage and releasing sites (mainly sarcoplasmic reticulum) in the cell are discussed. In addition, small arteries possess possibly more than three subtypes of alpha-adrenoceptors, including the presynaptic alpha 2-adrenoceptor. The roles of sarcoplasmic reticulum and the calcium receptor of contractile proteins (calmodulin or leiotonin C) from the chemically skinned muscles of vascular tissues were compared with those of intact muscles. The relaxation of vascular tissues as induced by activations of beta adrenoceptors, nitrites, and other chemicals is also briefly introduced. PMID- 6291412 TI - Angiotensin does contribute to drinking induced by caval ligation in rat. AB - In small (0.5 mg/kg) subcutaneous doses, the angiotensin-converting enzyme inhibitor, captopril, greatly enhanced drinking in response to caval ligation in the rat. Drinking was not secondary to urinary water loss since the rats developed a substantial positive fluid balance. High (50 mg/kg) subcutaneous doses of captopril reduced drinking to a level below that following caval ligation alone. This effect could be mimicked by giving repeated intracerebroventricular injections of captopril (total amount 110 micrograms) to rats treated with the lower subcutaneous dose of captopril. With this combination, therefore, not only did the lower dose enhancement disappear, the basal caval ligation drinking response was also reduced with a total dose of captopril of less than 2% of the higher subcutaneous dose alone. These results show that, when conversion of angiotensin I to angiotensin II is prevented in the brain as well as systemically, drinking in response to caval ligation is reduced although not entirely prevented. The original report that such drinking is multifactorial, depending on angiotensin as well as nonangiotensin mechanisms, is confirmed. PMID- 6291413 TI - Norwalk virus: a major cause of epidemic gastroenteritis. PMID- 6291414 TI - The frequency of a Norwalk-like pattern of illness in outbreaks of acute gastroenteritis. AB - Records of 642 outbreaks of acute gastroenteritis were reviewed to determine the proportion of outbreaks that were clinically and epidemiologically consistent with Norwalk-like virus infection. Using as our criteria stool cultures negative for bacterial pathogens, mean (or median) duration of illness 12-60 hours, vomiting in greater than or equal to 50 per cent of cases, and, if known, mean (or median) incubation period of 24-48 hours, we found that 23 per cent of waterborne outbreaks, 4 per cent of foodborne outbreaks, and 67 per cent, 60 per cent, and 28 per cent of outbreaks in nursing homes, in summer camps, and on cruise ships, respectively, satisfied the criteria for Norwalk-like pattern. Of 54 outbreaks that satisfied the criteria for Norwalk-like pattern, 14 were investigated for virus etiology. Ten of these (71 per cent) yielded serologic evidence of Norwalk-like virus infection. Norwalk-like viruses are probably an important cause of outbreaks of acute gastroenteritis. Investigation for Norwalk virus antibody in outbreaks that are clinically and epidemiologically consistent with Norwalk-like virus infection is likely to yield diagnostically useful results. PMID- 6291415 TI - Laboratory evaluation and assistance efforts: mailed, on-site and blind proficiency testing surveys conducted by the Centers for Disease Control. AB - During the last three years, the Centers for Disease Control (CDC) has conducted: 1) on-site surveys in which trained personnel visited laboratories that had experienced performance problems in the quarterly mailed proficiency testing (PT) program, reviewing the laboratories' analytical procedures by using carefully referenced samples to determine sources of errors and providing assistance in correcting them; 2) special assistance surveys in which carefully referenced samples were mailed to laboratories that had performed unsatisfactorily in routine mailed PT surveys and then telephone consultations were conducted to correct the problems; and 3) blind surveys in which carefully referenced samples were sent through normal patient sample acquisition routes to assess the actual day-to-day performance capability of the laboratories. Results suggest that on site surveys by trained laboratory surveyors and special mailed assistance surveys can be very effective in identifying the source of analytical errors in laboratories previously found, through mailed PT surveys, to have performance problems. Blind-survey results indicate that good performance in mailed PT does not necessarily imply good laboratory performance with routine patient specimens. Although difficult to conduct, blind surveys should be conducted whenever the logistics can be worked out by contractors for laboratory services, clinicians using laboratory services, and the laboratories themselves to assure the continuation of quality service. PMID- 6291416 TI - Epstein-Barr virus in normal pregnant women. AB - Acquired immune suppression accompanying normal pregnancy may be associated with reactivation of Epstein-Barr virus (EBV). Pregnant women with reactivated EBV having anti-EA antibodies show high titers of antiviral capsid antigen (VCA) geometric mean titers (GMT) of 522 versus 170 in those lacking anti-early antigen (EA). Among twenty-seven seropositive women at parturition, 17 (63%) had generated antibody to EA, and all 27 (100%) demonstrated significant increases in antibody to VCA (p less than 0.01). In contrast, antibody titers to cytomegalovirus, herpes hominis, varicella-zoster, and rubella viruses in the pregnant women were comparable to those found in nonpregnant controls. (Am J Reprod Immunol. 1982; 2:217-221.) PMID- 6291417 TI - Acute anterior cruciate ligament injury and repair reinforced with a biodegradable intraarticular ligament. Experimental studies. AB - The anterior cruciate ligament was transected at the femoral origin in the knee joint of 12 dogs. The ligaments were repaired in a conventional manner and reinforced with a polyglycolic acid (PGA) ligament of braided Dexon (Davis & Geck, Pearl River, NJ) suture. At two weeks the PGA ligament was still providing excellent support for the healing anterior cruciate ligament, and there was no synovitis within the knee joint. After five weeks, initial healing had firmly attached the repaired anterior cruciate ligament to the femoral condyle, and the PGA ligament had resorbed without inflammatory or fibrotic response. All repaired and reinforced ligaments in this series healed and provided functional stability to the knee joints. Biomechanical testing of the repaired anterior cruciate ligaments at four months produced a maximum strength of 54.2 +/- 6.3 kgf. Sulfur 35 uptake showed viable active collagen-producing cells in the repaired ligaments four months postoperatively. Thus, the biodegradable PGA ligament reinforced and splinted successfully the repaired anterior cruciate ligament. PMID- 6291418 TI - Operative ultrasonic features of insulinomas. AB - The technique of operative pancreaticosonography is described as a method of localizing occult insulinomas. The specific ultrasonic features of two small but palpable insulinomas are compared with those of other small lesions in the pancreas. Insulinomas are ultrasonically hypoechoeic and well-defined, with smooth borders, they deform but do not infiltrate surrounding structures, and they cause some dilatation of the secondary pancreatic ducts. Real-time, high resolution, intraoperative ultrasonic pancreatography is suggested as an adjunct to assist in the localization of insulinomas at surgery. PMID- 6291419 TI - [Indices of nucleic acid metabolism in the amniotic fluid in normal and pathological pregnancy]. PMID- 6291420 TI - [Role of para-influenza, respiratory-syncytial and adenovirus infections in pregnant women, fetuses and newborn infants]. PMID- 6291421 TI - [Androblastoma with heterologous elements]. PMID- 6291422 TI - Effects of hyperosmolality on the central nervous system and intracranial hemorrhage. AB - The relationship between hyperosmolality and intracranial hemorrhage as well as brain dysfunction was studied. Hyperosmolality was induced by continuous infusion of hyperosmotic solutions, such as 7% sodium bicarbonate (1,666 mOsm/1), 4.64% sodium chloride (1,666 mOsm/1), 20% glucose (about 1,200 mOsm/1), 20% mannitol (about 1,200 mOsm/1), and 10% glycerol with 0.9% sodium chloride and 5% fructose (about 2,000 mOsm/1). 1) Intracranial hemorrhage was observed without exception after the rabbits had died of hyperosmolality due to the infusion of these agents. The intracranial hemorrhage was mostly subdural hemorrhage in the subtentorial region. 2) The plasma osmolality just before death varied according to the agents administered and was lowest in the case of 7% sodium bicarbonate with a level of 441 mOsm/1. 3) The plasma osmolality in the case of 10% glycerol was higher than the case of 20% mannitol at the death. 4) The arousal reaction was suppressed by hyperosmotic solutions which contain sodium ions, and when the plasma osmolality surpassed 320 mOsm/1, dysfunction of the CNS began to appear. 5) The evoked muscular discharges caused by stimulation of the midbrain reticular formation were suppressed only by the administration of 7% sodium bicarbonate, so this result is related to alkalosis. PMID- 6291423 TI - Immunohistochemical and biochemical studies on collagen types in angiofibromas and shagreen patches from patients with tuberous sclerosis. AB - The distribution of collagen types in angiofibroma and shagreen patch from patients with tuberous sclerosis was examined biochemically and immunohistochemically by indirect immunofluorescence microscopy using type specific anti-collagen antibodies. Types I and III collagens were distributed in the interstitium of whole tumor tissues of angiofibriomas and a shagreen patch, showing relatively fine fibers at the periphery and large bundles in the center. No significant difference in the distribution of type I and III collagens was observed. Densitometric analysis of gel electrophoresis patterns of collagens solubilized from the skin by limited pepsin digestion revealed that both type I and III collagens were the main constituents of the diseased regions of the skin with tuberous sclerosis, and the ratio of type I collagen/type III collagen was relatively constant, which was not contradictory to the results of immunofluorescence microscopy. The presence of an unidentified component, probably type V collagen, was observed in the angiofibroma tissue from one patient with tuberous sclerosis. PMID- 6291424 TI - [Virological and serological survey of para-influenza virus in children of Rio de Janeiro]. PMID- 6291425 TI - Vesicular stomatitis in Brazil I--Isolation and identification of Alagoas strain. PMID- 6291426 TI - [Rotaviruses: an agent of infantile diarrhea]. PMID- 6291427 TI - [Vesicular stomatitis in Brazil. II - Epidemiological survey in equines, bats and Saguinus]. PMID- 6291428 TI - Study of some systematic errors during the determination of the total selenium and some of its ionic species in biological materials. PMID- 6291429 TI - [Simultaneous electronmicroscopic demonstration of different receptors at the surface of erythroid cells]. PMID- 6291430 TI - Electron microscope observations on preimplantation mouse embryos cultured with LiCl. AB - We have recently shown that LiCl in the culture medium retards cleavage of mouse preimplantation embryos without delaying their blastulation and causes the formation of blastocysts with few large cells and a reduced or absent inner cell mass (Izquierdo and Becker 1982). In this study we compare the ultrastructure of major cellular organelles of Li+-treated and control embryos. No subcellular alterations were found that correlate with the altered morphology of the blastocysts. On the basis of these results we submit that the malformation of blastocysts developed in a Li+-containing medium is the morphogenetic consequence of a retardation of cleavage coupled with a normal timing in the establishment of zonular tight junctions around the peripheral cells of the morula. PMID- 6291431 TI - Localization of enkephalin-like immunoreactivity in the cat carotid and aortic body chemoreceptors. AB - The purpose of this study was to determine if enkephalin-like immunoreactivity was present in the glomus cells of the carotid and aortic body peripheral arterial chemoreceptors. Cat carotid and aortic bodies were reacted with antisera to met- and leu-enkephalin using the indirect peroxidase-antiperoxidase immunocytochemical method of Sternberger (1979). Both the carotid and aortic bodies demonstrated clusters of immunoreactive cells for both met- and leu enkephalin. Additionally, met-enkephalin-like immunoreactivity was observed in many of the dense-core vesicles of the glomus cells of the carotid body. The glomus cells of these chemoreceptors are known to contain catecholamines which may modulate chemoreceptor activity. The presence of opioid peptide-like substances co-existing with the glomus cell catecholamines, perhaps in the same vesicles, may have important implications for a trophic influence of these peptides on glomus cell chemoreceptor modulation. PMID- 6291432 TI - Dissociation of plasma and cerebrospinal fluid beta-endorphin-like immunoactivity levels during pregnancy and parturition. AB - The association between central (cerebrospinal fluid [CSF]) and peripheral (plasma) levels of beta-endorphin-like immunoactivity (beta-ELI) in nonpregnant women (n = 8) and pregnant women (a) at 16 to 20 weeks of gestation (n = 6), (b) at term (n = 21), and (c) in labor (n = 15) was investigated. Umbilical arterial (n = 11) and venous (n = 11) samples were also obtained. In agreement with previous investigations, it was found that plasma levels of beta-ELI increased during labor (mean +/- SEM: nonpregnant women, 63.5 +/- 18.2; pregnant women at term, 64.0 +/- 12.2; women in labor, 110.8 +/- 30.3 pg/ml), and that levels of umbilical arterial plasma of beta-ELI exceeded those in umbilical venous plasma (132.5 +/- 34.0 versus 68.2 +/- 22.2). However, CSF levels of beta ELI did not change over the course of pregnancy or during labor (nonpregnant women, 36.5 +/- 15.8; pregnant women at 16 to 20 weeks of gestation, 60.1 +/- 10.3; pregnant women at term, 57.5 +/- 8.4; women in labor 48.5 +/- 8.3 pg/ml). This evidence that plasma and CSF levels of beta-ELI are dissociated during labor calls into question inferences regarding behavioral changes during parturition based on plasma beta-ELI measurements. PMID- 6291433 TI - Effects of furosemide on the neuromuscular junction. AB - These studies investigated the direct effects of furosemide on neuromuscular transmission using the in vitro rat phrenic nerve diaphragm and the in vivo at soleus nerve muscle preparations. Furosemide (10(-6)-10(-4)m) reduced the concentration of d-tubocurarine required to achieve 50% twitch tension depression in the indirectly stimulated rat diaphragm. Intraarterial injection of furosemide had a biphasic effect on the cat neuromuscular junction. At low doses (0.1-10.0 micrograms/kg) the drug had a depressant effect, reduced the force of muscle contraction, prevented nerve and muscle responses to NaF and dibutryl cyclic AMP, and intensified the neuromuscular blockade produced by d-tubocurarine and succinylcholine. In contrast, in higher doses (1-4 mg/kg) furosemide produced stimulus-bound repetitive neural activity, initiated neural activity, increased the force of muscle contraction, enhanced nerve and muscle responses to NaF and dibutyrl cyclic AMP, and antagonized d-tubocurarine and succinylcholine blockades. Furosemide had no effect on denervated preparations. High doses of furosemide inhibit non-competitively both the high- and low-affinity forms of the enzyme cyclic AMP phosphodiesterase in both soluble and particulate fractions of cat sciatic nerve. Thus, furosemide has direct effects on neuromuscular transmission, but the direction of these effects is dose-dependent. PMID- 6291434 TI - [Tactics of Arduan administration]. PMID- 6291435 TI - [Molecular mechanisms of the narcotic effect of general anesthetics]. PMID- 6291436 TI - [Glomus tumor. Clinical and diagnostic review, apropos of a case]. PMID- 6291437 TI - Gas mixing in the pulmonary airways. PMID- 6291438 TI - Estimation of distributions of ventilation/perfusion ratios. PMID- 6291439 TI - Quantitative computed tomography of vertebral spongiosa: a sensitive method for detecting early bone loss after oophorectomy. AB - We assessed serially the bone mineral loss in 37 premenopausal women for 24 months after oophorectomy and determined the dose-response for conjugated estrogen therapy in preventing this loss. Spinal cancellous bone was measured by quantitative computed tomography and measurement of appendicular cortical bone by radial photon absorptiometry and metacarpal radiogrammetry. For the placebo and low-dose treatment groups, the mean annual bone mineral losses were 7% to 9% from the vertebral spongiosum and 1% to 3% from the peripheral cortex. The correlation between axial and appendicular loss was weak (r = 0.581), precluding a reliable estimate of spinal loss from peripheral measurements. For the maximal-dose group (0.6 mg/d), the mean annual bone mineral losses were less than 0.5% from the axial and appendicular sites, and were not significant. The results indicate that spinal quantitative computed tomography provides a highly sensitive measurement of bone mineral loss after oophorectomy, that bone mineral loss is five- to sevenfold greater from the spinal spongiosum than from the appendicular cortex, and that conjugated estrogen in doses of less than 0.6 mg/d are inadequate to prevent the vertebral mineral loss. PMID- 6291440 TI - Infratemporal fossa approach for glomus tumors of the temporal bone. AB - The surgical experience with 74 glomus tumors of the temporal bone is presented. The article proposes new refinements in the classification of type C and D tumors and describes in detail the technique of blind sack closure of the external auditory auditory canal, double ligation of the sigmoid sinus and the use of a special infratemporal fossa retractor. The postoperative treatment and intraoperative management of the internal carotid artery, facial nerve and cranial nerves IX, X, XI and XII including the technique and results of a new type of V-VII anastomosis are discussed. PMID- 6291441 TI - Attempts to isolate bovine leukemia and bovine syncytial viruses from blood, uterine flush fluid, unfertilized ova and embryos from infected donor cattle. AB - Blood leucocytes, sediments of uterine flush fluid (UFF), eggs and embryos from 25 BLV-positive donor cows were tested for bovine leukemia (BLV) and bovine syncytial (BSV) viruses by cocultivation with fetal lamb spleen cells and by applying syncytium induction and immunofluorescence tests. BLV was diagnosed in 11/15 (73.3%) leucocyte and 4/25 (16.0%) UFF-sediment specimens as compared to BSV in 14/15 (93.3%) and 21/25 (84.0%) of the similar specimens and neither BLV or BSV were found in 26 eggs and 60 embryos collected from 20 of the 25 cows. Detection of BLV antigens by immunofluorescence was hampered by the competitive replication of both BLV and BSV and competitive growth in indicator cells and uterine cells. As BLV has not been observed in cells of UFF sediments, it was probably isolated from leucocytes present in the lumen of uterus or from blood seeping out from inapparent vessel damage during flushing. Isolation of BLV in UFF sediments gives additional evidence to the concept of a transplacental transmission by a not yet elucidated mechanism. The high rate of BSV recovery from cells of UFF sediments indicated that this virus is more wide-spread than previously shown and that it may play a role in causing disorders of the reproductive tract. PMID- 6291442 TI - [Frequency of enteropathogenic K99+ ST+ Escherichia coli and rotaviruses in neonatal diarrhea of calves. Survey of a veterinarian's clientele in Sarthe]. AB - A clinical and microbiological study was carried out on 21 diarrhoeic calves less than 20 days old, belonging to 20 different farms in the French department of Sarthe. Each diarrhoeic calf was compared with a clinically healthy calf of the same age, from the same farm. Two visits were made : one during the acute phase of diarrhoea, the other 10 to 15 days later. Of the 21 diarrhoeic calves, nine died. On the first visit, rotavirus was found in faeces of 11 diarrhoeic calves and E. coli K99+ ST+ in faeces of three diarrhoeic calves, in two cases together with rotavirus. At the same time, rotavirus was found in the faeces of five control calves whereas E. coli K99+ ST+ was not present. On the second visit, rotavirus was detected in the faeces of one of the 12 surviving diarrhoeic calves and in the faeces of two control calves. No E. coli K99+ and ST+ was found in any of the two groups. Short duration antibiotic treatment (sulfonamide + colistin) carried out on the first visit did not result in increased resistance of E. coli to the ten antibiotics tested. On the contrary, cases of resistance to certain of these antibiotics were more frequent during the acute phase of illness (and before any treatment) than 10 to 15 days later. This fact was explained by the presence, in at least five cases, of multiresistant E. coli strains which disappeared with recovery. PMID- 6291443 TI - Influence of tumor burden on red blood cell deformability in small cell lung cancer patients. AB - Red blood cell deformability was measured in 54 newly diagnosed patients with small cell bronchogenic carcinoma and in 22 normal hospital employees. Twenty-one small cell cancer patients had disease confined to one hemithorax plus draining lymph nodes (one organ system), 15 had involvement of one additional metastatic site (two organ systems), 12 had two metastatic sites (three organ systems) and 6 patients had 4 organ systems involved. Patients with one or two organ system involvement had red cell deformability results comparable to the normal control group. Patients with tumor extension to 3 or 4 organ systems had significantly decreased deformability relative to patients with lesser tumor burdens. Based on the lowest deformability results in the control population small cell cancer patients were divided into those with normal or impaired red blood cell deformability. The latter patients were significantly more anemic and had higher red cell mean corpuscular volumes than their normal counterparts. Patients with impaired deformability also had significantly elevated blood glucose levels and significantly lower serum inorganic phosphorus and iron binding capacity. Since red cell deformability is the principal determinant of capillary blood flow abnormal deformability may result in decreased tumor oxygenation and decreased, and non-uniform, delivery of chemotherapeutic agents to tumor tissue. PMID- 6291444 TI - Infectious mononucleosis in Singapore. PMID- 6291445 TI - Further evaluation of diethyldithiocarbamate as an antagonist of cisplatin toxicity. PMID- 6291446 TI - [Metabolism of insulin and immunoreactive insulin-like substances (proinsulin, HWIRI) in vivo in man]. AB - Immunoreactive insulin (IRI) and its three constituents as determined by gel filtration: high molecular weight substances with insulin immunoreactivity (HWIRI), proinsulin (PI) and insulin (I), were measured hourly during 28 hours in the portal, hepatic and peripheral venous systems of a patient operated for a nesidioblastoma. The PI levels were constant from one system to the other (7.1 +/ 0.8 microU/ml; 7.3 +/- 0.8 microU/ml; 9.2 +/- 1.2 microU/ml). The I levels decreased from the portal (53.3 +/- 5.2 microU/ml) to the hepatic venous system (28.0 +/- 1.0 microU/ml) and from the latter to the peripheral venous system (19.8 +/- 2.0 microU/ml). The HWIRI levels were stable between the portal and the hepatic venous systems (4.7 +/- 0.7 microU/ml; 4.8 +/- 0.6 microU/ml) and higher in the peripheral system (8.2 +/- 1.2 microU/ml). Our results demonstrate that: - I is essentially degraded by the liver. --There is no variation of PI in the three compartments. --HWIRI is formed in the circulation. --Variations of IRI levels between venous systems acquire physiological significance only when accompanied by the variations of the levels of each constituent. PMID- 6291448 TI - Peripheral neuropathy, plasma cell dyscrasia, and hot blood. PMID- 6291447 TI - Peripheral nervous system complications of coronary artery bypass graft surgery. AB - Among 421 patients undergoing coronary artery bypass graft surgery, 55 (13%) developed 63 new peripheral nervous system (PNS) complications postoperatively. Most common was a brachial radiculoplexopathy, which occurred in 23 patients. Of these, 21 involved lower trunk or medial cord fibers. In 17 there was a correlation between the site of jugular vein cannulation and the side affected, suggesting that needle trauma played a role. Stretching from chest wall retraction may have caused some cases. Other deficits included 13 saphenous, 8 common peroneal, and 5 ulnar mononeuropathies. Six patients had persistent singultus, suggesting phrenic nerve involvement. Unilateral vocal cord paralysis was found in 5. An isolated partial Horner syndrome and a facial neuropathy were also identified. Males were more likely to develop PNS complications. Hypothermia during surgery was associated with increased risk. Most PNS deficits were transient, and lasting disability was rare. PMID- 6291449 TI - [Mechanism of polyene antibiotic inactivation. Changes in the physicochemical characteristics and the destruction of the polyene chromophore levorin during inactivation]. AB - The results of studying free radicals of some polyenic antibiotics with the EPR method are presented. It is shown that the number of free radicals increased by 100 per cent with a 2-fold decrease in the biological activity. A qualitative change in the EPR spectrum due to the presence of a new radical type was also observed. The changes in the spectral characteristics of levorin allowed one to demonstrate that breaking of the links and formation of the polymer products during oxidative destruction of the polyenic chromophore resulted from attachment of the radicals through the antibiotic double bonds. The data correlate with the results of the quantum-chemical evaluation of the polyenic chromophore. PMID- 6291450 TI - [Separation and characteristics of the components of the antibiotic virenomycin]. AB - The composition of virenomycin, a new antitumor antibiotic was studied. Two components V and M were detected with high resolution liquid chromatography and thin layer chromatography on siluphol (Czechoslovakia) and silica gel (Merk, BRD). A preparative method for separation of the antibiotic components with the use of chromatography on columns with silica gel was developed. Biological and physicochemical properties of separate components were studied to show that they significantly differed by their antibacterial action in vitro: virenomycin V was 2 to 4 times more active than virenomycin M against a number of microbes. The physicochemical properties of the components are similar. It was shown with mass spectrometry that the molecular weight of virenomycin is 12 units higher than that of virenomycin M. The PMR spectra showed that this difference is due to the presence of a vinyl group in the chromophore moiety of the virenomycin V molecule and a methyl group at the similar site of the virenomycin M molecule. PMID- 6291451 TI - Activity of cefoperazone and two beta-lactamase inhibitors, sulbactam and clavulanic acid, against Bacteroides spp. correlated with beta-lactamase production. AB - A total of 102 isolates of Bacteroides spp. were studied for beta-lactamase production and susceptibility to cefoperazone alone or in combination with either of the beta-lactamase inhibitors sulbactam and clavulanic acid. The geometric mean minimal inhibitory concentration of cefoperazone alone was 31.5 micrograms/ml and when combined with 10 micrograms of sulbactam per ml or 2 micrograms of clavulanic acid per ml was reduced to 5.4 and 9.2 micrograms/ml, respectively. When bacterial suspensions were tested for beta-lactamase production with nitrocefin, 91 (89.2%) of these isolates produced the enzyme. The geometric mean minimal inhibitory concentrations of cefoperazone rose only slightly for isolates with low or intermediate enzyme activity but rose significantly for those with high activity. The addition of EDTA to cefoperazone significantly more frequently enhanced the activity of cefoperazone against beta lactamase-negative as opposed to beta-lactamase-positive isolates. Furthermore, EDTA resulted in synergistic activity of the cefoperazone-sulbactam combination on beta-lactamase-positive isolates for which the combination had previously not shown a synergistic effect. This study demonstrates the relationship between beta lactamase production and the resistance of Bacteroides spp. to cefoperazone and shows that inhibition of these enzymes can reverse this resistance. PMID- 6291452 TI - Capacity of deoxycytidine to selectively antagonize cytotoxicity of 5-halogenated analogs of deoxycytidine without loss of antiherpetic activity. AB - Enzyme kinetic studies from this laboratory (M. Dobersen and S. Greer, Biochemistry 17:920-928, 1978) suggested that deoxycytidine could antagonize the toxicity of 5-halogenated analogs of deoxycytidine without interfering with their antiviral activity. Antagonism by deoxycytidine of the toxicity of 5 chlorodeoxycytidine without impairing its anti-herpes simplex virus type 2 activity is demonstrated in the present studies. Tetrahydrouridine, an inhibitor of cytidine deaminase, was utilized. The high Km for deoxycytidine (0.6 mM) with respect to the herpes pyrimidine nucleoside kinase as compared with the low Km for 5-chlorodeoxycytidine (1.1 microM) accounts for the absence of antagonism of the antiviral activity. The high Km for 5-chlorodeoxycytidine (56 microM) as compared with the low Km of deoxycytidine (2 microM) with respect to mammalian deoxycytidine kinase accounts, in great part, for the antagonism of toxicity. In addition, antagonism of toxicity by deoxycytidine is the result of factors other than the kinetic parameters of nucleoside kinases, as indicated by its antagonism of the cytotoxicity of 5-chlorodeoxyuridine. This may be attributed to replenishment of low dCTP pools, diminished because of effector inhibition of ribonucleoside diphosphate reductase by Cl-dUTP. Resistance of the herpes-encoded enzymes to effector control may also play a role in the selective antagonism. Cell culture studies with high concentrations of tetrahydrouridine and 2' deoxytetrahydrouridine suggest that competition by deoxycytidine for deaminases may not play a major role. The fact that deoxycytidine antagonizes the toxicity of chlorodeoxyuridine also argues against competition for the deaminases as a major reason for its effect. Limited studies with a topical herpes simplex virus type 2 infection system indicate heightened efficacy of 5-chlorodeoxycytidine (and tetrahydrouridine) when deoxycytidine is coadministered. The concepts of selective antagonism of a chemotherapeutic agent derived from these studies may be applied to other approaches that extent beyond viral chemotherapy. PMID- 6291453 TI - Action of antifungal imidazoles on Staphylococcus aureus. AB - In Staphylococcus aureus, using the imidazoles miconazole and ketoconazole, detailed studies of minimal inhibitory concentrations, kinetics of growth, viability, and release of intracellular K+ confirm that the two imidazoles work differently in this bacterium. Miconazole is bactericidal at low concentrations and causes release of cellular K+. Ketoconazole has no bactericidal effect at any tested concentration and has little effect on K+ permeability of S. aureus; it slows growth at high concentration. This is reflected in a low minimal inhibitory concentration for miconazole and a high one for ketoconazole. The probable mechanisms of the bacteriostatic and bactericidal effects of the imidazoles are discussed in light of these results and the previously described antifungal mechanisms of the drugs. alpha-Tocopherol blocks the action of both imidazoles. PMID- 6291454 TI - Synergistic activity of mecillinam in combination with the beta-lactamase inhibitors clavulanic acid and sulbactam. AB - The beta-lactamase inhibitors clavulanic acid and sulbactam were combined with mecillinam. beta-Lactamase-containing Escherichia coli resistant to mecillinam was synergistically inhibited by both clavulanic acid and sulbactam. beta Lactamase-containing Enterobacter was synergistically inhibited, but strains lacking beta-lactamases were not synergistically inhibited. Synergistic inhibition was noted for beta-lactamase-containing, mecillinam-resistant Klebsiella, Citrobacter, Serratia, and Salmonella isolates, but only 18% of beta lactamase-containing Proteus mirabilis, Providencia rettgeri, Providencia stuartii, and Morganella morganii were synergistically inhibited by the combinations. PMID- 6291455 TI - Rifampin inhibition of the occluded virus form of a nuclear polyhedrosis virus. AB - Rifampin at concentrations toxic to noninfected cells but not to infected cells is a selective inhibitor of occluded virus of the group A Baculoviridae (nuclear polyhedrosis virus). However, the titer of nonoccluded virus is not affected. Rifampin blocks occlusion until late in the replication cycle (14 to 16 h), and its effects are reversible. Modes of action of polyhedral inclusion body production are unknown. PMID- 6291456 TI - Mode of action of acyclovir triphosphate on herpesviral and cellular DNA polymerases. AB - The effect of 5'-triphosphate of acyclovir (ACV) on DNA polymerases of two human herpes-viruses, herpes simplex virus type-1 (HSV-1) and Epstein-Barr virus (EBV) as well as human cellular DNA polymerases alpha and beta has been examined. Of the enzymes tested, HSV-1 DNA polymerase was the most sensitive to inhibition by acyclovir triphosphate (ACVTP). The EBV DNA polymerase and DNA polymerase beta were less sensitive. ACVTP inhibition was competitive with dGTP with Ki values of 0.03, 0.15, 9.8 and 11.9 microM for HSV-1 DNA polymerase, DNA polymerase alpha, EBV DNA polymerase and DNA polymerase beta, respectively. Substituting a synthetic primer template (dG) approximately 15 x (dC)n for activated DNA template did not alter the pattern of inhibition. In a time course experiment, addition of ACVTP instead of dGTP did not increase DNA synthesis and it appeared to act as a chain terminator in DNA replication catalyzed by either HSV-1 DNA polymerase or DNA polymerase alpha. Although EBV DNA polymerase was less sensitive to ACVTP inhibition, the nucleoside analog itself was inhibitory to EB virus production by P3HR1 cell line as determined by a reduction in the percentage of cells expressing virus capsid antigen (VCA). On day 4, ACV at 10 and 25 micrograms/ml reduced the cell growth by 10% and 32%, respectively, while it reduced the VCA-positive cells by 80% and 84%, respectively. These results indicate that inhibition of EBV DNA polymerase activity by ACVTP may not be the primary mechanism responsible for ACV inhibition of EBV replication. PMID- 6291457 TI - Recurrent herpes simplex in mice; topical treatment with acyclovir cream. PMID- 6291458 TI - Cholecystokinin-octapeptide effects on eating elicited by "external" versus "internal" cues in rats. PMID- 6291459 TI - [Hepatitis virus A antibodies (anti-HA) in various population groups of the Antwerp area]. PMID- 6291460 TI - The effect of hyperoxia on superoxide production by lung submitochondrial particles. PMID- 6291461 TI - Stoichiometry of phosphorylation of hepatic ATP-citrate lyase by protein kinase. PMID- 6291462 TI - Effects of divalent cations on the activity of ribulosebisphosphate carboxylase: interactions with pH and with D2O as solvent. PMID- 6291463 TI - Evidence for the intestinal type of fructose 1,6-bisphosphatase in mouse, rat, and golden hamster. PMID- 6291464 TI - Characterization of 1,25-dihydroxyvitamin D3-receptor complex interactions with DNA by a competitive assay. PMID- 6291465 TI - Conservation of primary structure at the proteinase-sensitive site of fructose 1,6-bisphosphatases. PMID- 6291466 TI - [Association of a gastric epithelioid leiomyosarcoma and a functioning extra adrenal paraganglioma. A new case suggestive of a single syndrome]. PMID- 6291467 TI - [Personal experience with the application of the electron microscope in research on Rotavirus in the feces of children with acute gastroenteritis]. PMID- 6291468 TI - [Effect of culture conditions on the aging of RK13 cells used for the titration of vesicular stomatitis virus]. PMID- 6291469 TI - [Oncogenicity of type C retroviruses]. PMID- 6291470 TI - [Effect of inhibition of converting enzyme by captopril on arterial pressure, renin and aldosterone in essential hypertension]. AB - The response of arterial blood pressure, plasma renin activity (PRA) and plasma (PA) or urinary aldosterone (UA) concentrations to the administration of captopril, was studied in patients with established essential hypertension. Captopril was effective in lowering significantly the blood pressure (189.4/111.2 +/- 23.9/9.7 to 163.4/98.1 +/- 20.7/8.6 mmHg. mean +/- SD, p less than 0.01/0.001). Normal arterial blood pressure values (140.4/86.5 +/- 20.7/10.8 mmHg, were achieved by the addition of hydrochlorothiazide. Captopril administration was followed by a decrease in PA and in UA and an increase in PRA, suggesting the inhibition of angiotensin II formation. Captopril attenuated hypokalemia and hyperaldosteronism produced by the simultaneous administration of hydrochlorothiazide. PMID- 6291471 TI - Binding properties of 3H-lofentanil at the opiate receptor. PMID- 6291472 TI - Halopemide and benzodiazepine binding sites. AB - Halopemide inhibits 3H-BZ binding to crude and washed rat forebrain membranes with IC50 values of 16-25 microM. Its putative metabolites are considerably less active. The actions of halopemide are probably not directly related to its ability to interfere with high-affinity 3H-GABA binding sites. However, halopemide may have some unique properties in this regard. PMID- 6291473 TI - Solubilized opiate receptors labelled with 3H-lofentanil. PMID- 6291474 TI - Sidedness of plasma membrane vesicles isolated from intestinal smooth muscle. PMID- 6291475 TI - Ketoconazole blocks testosterone synthesis. AB - Ketoconazole, a new oral drug used to treat systemic and superficial mycoses, inhibits sterol synthesis in fungi. The development of gynecomastia in two patients prompted us to investigate the effect of the drug on testosterone production. After a 200-, 400-, or 600-mg dose, volunteer male testosterone serum concentrations fell markedly, but returned toward baseline eight to 24 hours later as ketoconazole serum concentrations waned. A marked but transient drop in testosterone levels occurred in patients receiving long-term therapy, and continuous testosterone depression was noted in one. A block of synthesis was demonstrated in vitro. Ketoconazole at concentrations achievable in serum with currently used doses blocked basal and gonadotropin-stimulated testosterone production by rat Leydig cells. The diminution of testosterone synthesis could be significant as further therapeutic trials may use larger doses or more than once daily administration. The paucity of reports of endocrinologic toxicity may relate to the "escape" from the block demonstrated in vivo. PMID- 6291476 TI - Intracellular mechanism of photoreceptor excitation. PMID- 6291477 TI - Processing of visual signals in vertebrate photoreceptors. AB - Photoreceptors of the vertebrate retina hyperpolarize in response to illumination. The conductance changes in the plasma membrane associated with the electrical response are the final step of chain of events initiated by light absorption at the outer segment of the visual cell. The mechanism whereby the free energy of photons in converted into neural information is largely unknown. Present knowledge is consistent with the idea that an internal transmitter is modulated by light and modifies the ionic permeability of the plasma membrane. As to the identity of the internal messenger two candidates have been proposed: Ca2+ and cyclic GMP respectively. Increasing evidence suggests that both substances may be involved in the process of phototransduction. The electrical response of photoreceptors does not simply reflect the light absorbed by the cell: complex interactions occurring between adjacent photoreceptors and between photoreceptors and second order neurons cooperate with the initial process in determining the final shape of the receptor message. Recurrent interactions involve particularly cones: their membrane potential can be modified at least by three distinct mechanisms; i) by light absorption at their outer segment; ii) by light absorption at the outer segment of neighbouring cones, and iii) by potential changes occurring in horizontal cells. PMID- 6291478 TI - [Inclusion body enteritis in pigeons caused by adenovirus and parvovirus]. PMID- 6291479 TI - [Morphology, maturation and activity of duck pest virus in host tissue--an electron microscopic study]. PMID- 6291480 TI - [Effect of various culturing conditions on the release of bovine leukosis virus from peripheral leukocytes in cattle]. PMID- 6291481 TI - [Effect of mitogens on the release of cattle leukosis virus by short-term cultured bovine leukocytes]. PMID- 6291482 TI - [Routine determination of virus concentrations by density gradient centrifugation and ultraviolet analysis. 1. Methodological experiences]. PMID- 6291483 TI - [Electron microscopic virus demonstration in the McCoy cell line--virus morphogenesis in ultrathin sections]. PMID- 6291484 TI - [Determination of the quantity of minimal antigen for oral immunization of sows against transmissible gastroenteritis of swine using Riems TGE vaccine]. PMID- 6291485 TI - [Efficacy testing of inactivated rotavirus vaccine in a dairy cattle herd]. PMID- 6291486 TI - [Radioimmunoassay for demonstration of FMD-specific antibodies in sera of experimentally infected animals]. PMID- 6291487 TI - Periductal elastic tissue of breast cancer. Quantitative histologic study. AB - Using techniques of stereology, we estimated the relative volume of periductal elastic tissue in 60 infiltrating ductal carcinomas of the breast. Volume density of periductal elastic tissue in the neoplasm correlated with our own subjective histologic estimates of the amount of elastic tissue. Periductal elastic tissue had a significantly higher volume density in the neoplasm than in nonneoplastic breast, but the two estimates did not correlate with each other. Similarly, the volume density of elastic tissue in the neoplasm did not correlate with that of neoplastic cells or of stromal collagen in the neoplasm, degree of lymphocytic infiltration, lymph node metastasis, mortality, menstrual status, age, parity, or presence or levels of estrogen receptor protein in the neoplasm. On the other hand, parity correlated with the volume density of periductal elastic tissue in the nonneoplastic parenchyma of the breast. Our findings indicate that there are at least two separate elastogenic effects in infiltrating ductal carcinoma of the breast: that exerted by parity on nonneoplastic tissue and that exerted by cancer itself on the neoplasm. PMID- 6291488 TI - 'Mixed' salivary type adenoma of the human female breast. AB - A breast lump in a 46-year-old woman proved on histologic examination to be an adenoma of the "mixed" salivary type (pleomorphic adenoma). The lesion was characterized by intense adenomatous proliferation, hyperplasia of epithelial and myoepithelial cells, myxoid areas, and focal squamous differentiation. Electron microscopy confirmed the myoepithelial nature of islands of stromal cells. The microanatomic pattern, histochemical staining reactions, the presence of elastin, and the ultrastructural features of the tumor, all confirmed the close similarity to the corresponding salivary gland neoplasm. PMID- 6291489 TI - Two ovarian malignant neoplasms. PMID- 6291490 TI - alpha 1-Antitrypsin deficiency and hepatocellular carcinoma. Association with cirrhosis, copper storage, and Mallory bodies. AB - A pathogenetic relationship between hepatic manifestations of alpha 1-antitrypsin deficiency (AATD) and hepatocellular carcinoma has previously been suggested but not established. We encountered three patients with hepatocellular carcinoma (HCC) and nonneoplastic hepatocytes containing globules positive for alpha 1 antitrypsin by immunoperoxidase staining or immunofluorescence microscopy. The AATD was severe in one patient, mild in the second, and not evaluated in the third; any pathogenetic relationship between the deficiency and HCC must therefore remain speculative. Nonneoplastic hepatocytes of all three patients demonstrated copper storage and, in two patients who were not alcoholic, Mallory bodies. Simultaneous presence of copper storage and Mallory bodies has not been reported in liver disease associated with AATD. The pathogenetic mechanisms of these findings are unknown. PMID- 6291491 TI - Peripheral neuropathy after concomitant dimethyl sulfoxide use and sulindac therapy. AB - The case is presented of a 63-year-old man with a long history of degenerative arthritis who took sulindac (Clinoril) 200 mg BID for 6 months with no untoward effects. Then, without physician knowledge, he began using 90% dimethyl sulfoxide (DMSO) topically to his upper and lower extremities. Shortly thereafter, he developed a profound mixed sensorimotor peripheral neuropathy. Serial electromyographic and nerve conducion studies performed at intervals of several months for 1 year suggested both segmental demyelination and axonal neuropathy. The patient experienced initial deterioration followed by gradual but incomplete recovery. PMID- 6291492 TI - Arterial regeneration over absorbable prostheses. AB - Small-vessel arterial replacement by a prosthetic material elicits inflammatory and regenerative reactions that alter the patency rate of the resultant prosthesis-tissue complex. Woven absorbable polyglycolic acid prostheses, 24 mm in length, were implanted into adult rabbit aortas and followed for 7 1/2 months. At the time the animals were killed, 45 specimens were studied grossly and by arteriography, light and electron microscopy, immunohistology, and bursting strength determinations. No rabbit died of aortic-related complications. Mild dilation occurred in 11% (five specimens), and intimal hyperplasia occurred in 13% (six specimens), with 76% (34 specimens) displaying parallel walls at the original 3- to 4-mm diameter. Microscopy showed that the prostheses were replaced by regenerating endothelialized vessels containing smooth musclelike myofibroblasts and dense fibroplasia, without thrombosis or infection. All withstood saline solution infusion at three to five times the systolic pressure. Results of these studies demonstrate conditions that permit arterial regeneration over absorbable scaffolds. PMID- 6291493 TI - [Electromechanical effects of polymyxin B on the isolated atrium and heart in situ]. PMID- 6291494 TI - Changes in collagen metabolism in diseased muscle. I. Biochemical studies. AB - Possible changes in collagen biosynthesis were studied in 50 patients with neuromuscular disorders and 14 controls. Type III procollagen aminoterminal propeptide concentrations and galactosylhydroxylysyl glucosyltransferase (GGT) activities were assayed in serum, and prolyl 4-hydroxylase and GGT activities were assayed in muscle biopsy specimens. All four assays showed significantly elevated values in cases of polymyositis, adult forms of muscular dystrophy, and amyotrophic lateral sclerosis, the concentration of muscular collagen also being significantly increased in the last two conditions. Some abnormalities were also seen in polyneuropathy, myotonia congenita, and undefined myopathy. High correlations were found among the values for the four assays, but no marked correlations with muscular collagen concentration or enzyme activities characteristic of neuromuscular disorders were found. The four assays may reflect changes in actual collagen synthesis in the diseased muscle. PMID- 6291495 TI - Inclusion body myositis. A corticosteroid-resistant idiopathic inflammatory myopathy. AB - In seven patients with slowly progressive muscle weakness, inclusion body myositis (IBM) was diagnosed on biopsy. None had stigmata of collagen-vascular disease or malignancy. Serum creatine kinase levels were mildly or moderately increased. The six patients treated with prednisone did not improve. Needle electromyography showed a "myopathic" pattern in all patients, but four also had diffuse neurogenic changes with normal nerve conductions. Histologic study of muscle showed a mixture of small rounded fibers varying in size, atrophic angulated fibers forming small groups, and hypertrophic fibers. Variable amounts of inflammation, necrosis, and regeneration were seen in all specimens. All showed numerous intracytoplasmic vacuoles lined with purple-blue granules. Electron microscopy showed membranous whorls and masses of abnormal filaments measuring 14 to 18 nm in diameter. Although IBM seems to be a distinct type of inflammatory myopathy, its etiology and pathogenesis are not clear. PMID- 6291496 TI - CNS varicella-zoster vasculitis. PMID- 6291497 TI - Combined ocular infection with cytomegalovirus and cryptococcosis. AB - To our knowledge, this is the first published description of the clinical and pathologic features of combined ocular infection with both cytomegalovirus and Cryptococcus. Infection occurred in a patient treated by immunosuppression and corticosteroid therapy. This article emphasizes that ocular findings may be the first sign of disseminated opportunistic infection and that clinical manifestations may be sufficient to allow accurate diagnosis of opportunistic retinitis. PMID- 6291498 TI - Importance of bicarbonate ion in the vitreous space. AB - Intraocular irrigating solutions used during vitrectomy must be safe for all tissues surrounding the vitreous space. Bicarbonate and glucose were essential to the maintenance of retinal function. In this study the optimum concentration of bicarbonate in the irrigating solution for the retina was determined electrophysiologically with the use of the in vitro eyecup of the rabbit. In 15 to 25 mmole/L of bicarbonate solutions, the electroretinographic (ERG) b wave was well maintained. Higher concentrations of bicarbonate (35 to 45 mmole/L), which have been considered to be advantageous for the integrity of the corneal endothelium, were shown to suppress the retinal activity. These results were also found in in vivo eyecup, as determined with ERG and visual evoked response. Properly performed open-sky vitrectomy, itself, does not cause severe damage to the normal retinal function. PMID- 6291499 TI - Three modes of transmission of Ross River virus by Aedes vigilax (Skuse). AB - Some 15% of the saltmarsh mosquito, Aedes (Ochlerotatus) vigilax (Skuse) mechanically transmitted the T48 strain of Ross River virus from viraemic donors to uninfected recipient suckling mice. The population tested from near Brisbane was moderately susceptible to infection (ID50=10(4X5+/-0X1) suckling mouse LD50/mosquito) when fed virus diluted in blood on a cotton pledget. Rapid proliferation of the virus in the mosquito allowed transmission from 4 days after ingestion with maximum transmission of 50-57% occurring from 10 days. From assay of Ae vigilax ovaries and of progeny reared from parentally-infected females, preliminary data suggest transovarial transmission as a potential survival method for this virus. PMID- 6291500 TI - A survey of serum antibody to avian haemagglutinating adenovirus in Queensland poultry flocks. PMID- 6291501 TI - [Determination of enzyme activity in horse organs]. PMID- 6291502 TI - [EHV-1 myeloencephalitis in the horse]. PMID- 6291503 TI - [Canine insulinoma. Clinical and morphological findings]. PMID- 6291504 TI - Locus coeruleus - cerebellum: interaction during development. AB - The presentation describes a combined morphological and biochemical analysis of the developmental interaction between the locus coeruleus (LC) and the cerebellum of rats, which receives an afferent innervation from the LC. The LC neurons are among the first CNS neurons to arise during ontogeny. They establish axonal connections to their target areas while migrating into their nuclear area, where they collect around E17. Their perikaryal development proceeds through the well known stages of neuronal differentiation, Nissl body formation as a sign of synaptic connectivity appearing for the first time on E18. However, changes in the LC-perikarya are taking place in early postnatal stages. Perikaryal volume increases to reach a transitory maximum of 150% of the adult value on P15. Ultrastructurally, a dissolution of Nissl bodies and an increase in the number of polyribosomes are seen during this developmental period, reminiscent of perikaryal changes during the axon reaction. Later, the organization of ribonucleoproteins into Nissl bodies is re-established. NA axons are demonstrable in the cerebellar anlage for the first time on E17. They increase rapidly in number and organization during cerebellar development as shown by catecholamines histofluorescence. Quantitative measurements of cerebellar high-affinity uptake for NA show that a peak of NA innervation is reached on P10, which amounts to about 250% of the adult value. This hyper-innervation is transitory and declines to adult values on P20. The period of cerebellar NA hyperinnervation corresponds to the perikaryal changes in volume and ultrastructure of LC neurons. The phenomenon of transitory hyperinnervation of a target area is discussed with respect to the establishment of axonal connections during normal development and in regeneration. PMID- 6291505 TI - Response of central monoamine neurons following an early neurotoxic lesion. AB - Systemic treatment with the selective monoamine neurotoxins 6-hydroxydopamine (6 OH-DA), N-(2-chloroethyl)N-ethyl-2-bromobenzylamine (DSP4) or 5,7 dihydroxytryptamine (5,7-HT) in the neonatal stage produces marked and permanent alterations of the postnatal development of central noradrenaline (NA) and 5 hydroxytryptamine (5-HT) neurons. 6-OH-DA and DSP4 act preferentially on NA neurons, in particular on the locus coeruleus system, whereas 5,7-HT acts on 5-HT neurons. The neurotoxin treatment leads to pronounced denervations of distant nerve terminal projections while innervation areas close to the cell bodies become hyperinnervated. The total number of monoamine nerve terminals developed in the CNS after the neurotoxin treatment is approximately unchanged. A surgical lesion of NA and 5-HT axons in the neonatal stage causes similar changes compared to a neonatal neurotoxin treatment. The postsynaptic monoamine receptors appear to develop independently of the presynaptic nerve terminals, although the transmitter availability at the receptors is able to regulate the number of receptors (up and down regulation). The transmitter turnover is increased in terminals spared by the neurotoxin in denervated areas, while decreased in hyperinnervated regions. The alterations in receptor density and transmitter turnover may represent compensatory mechanisms. Substance P has a counteracting effect while morphine has a potentiating effect on the 6-OH-DA-induced NA denervation and hyperinnervation, indicating that the functional state of the neuron may modulate the final outcome of a neonatal 6-OH-DA treatment. The results indicate that the altered development of central monoamine neurons following a neonatal monoamine neurotoxin treatment or axotomy is mainly related to a 'pruning effect', i.e. the prevention of the development of one axonal branch leads to a proliferative growth response in intact branches. PMID- 6291507 TI - Does superoxide anion participate in 2-oxoglutarate-dependent hydroxylation? AB - The possible role of superoxide anion in 2-oxoglutarate-coupled dioxygenase reactions has been investigated. gamma-Butyrobetaine hydroxylase (EC 1.14.11.1) was inhibited by human erythrocyte superoxide dismutase (EC 1.15.1.1), probably due to release of Cu(2+) or Zn(2+), as the inhibition was more pronounced after heat-inactivation of the dismutase and as Cu(2+) was a potent inhibitor. Bovine superoxide dismutase and the Mn(2+)-containing superoxide dismutase from Escherichia coli were not inhibitory. Superoxide anion generated from xanthine/xanthine oxidase was not stimulatory and could not replace ascorbate. Thymine 7-hydroxylase (EC 1.14.11.6) and thymidine 2'-hydroxylase (EC 1.14.11.3) were not inhibited by erythrocyte superoxide dismutase or stimulated by superoxide anion. gamma-Butyrobetaine hydroxylase was inhibited by a number of low-molecular-weight compounds, such as tetranitromethane, Nitro Blue Tetrazolium, adrenaline and Tiron, which may act as scavengers of superoxide anion. Involvement of this radical in other oxygenase reactions has been inferred from the findings that they were inhibitory for the respective enzymes. Several of these compounds also inhibited gamma-butyrobetaine hydroxylase. It could be concluded from these experiments, however, that mechanisms other than disposal of superoxide anion might equally well be operative, such as hydrophobic interaction with the enzyme protein and interaction with compounds required for full enzymic activity, e.g. iron and ascorbate. The results appear to rule out a requirement for superoxide anion generated in free solution, and have not yielded evidence for participation of enzyme-bound superoxide anion in 2-oxoglutarate-dependent hydroxylations. PMID- 6291506 TI - Organization of alpha-chain genes among Hb G-Philadelphia heterozygotes in association with Hb S, beta-thalassemia, and alpha-thalassemia-2. PMID- 6291508 TI - Isolation of a 5.3-S calmodulin-deficient 3':5'-cyclic nucleotide phosphodiesterase from cardiac muscle. AB - 1. In the presence of Ca2+, a 5.3-S 3':5'-cyclic nucleotide phosphodiesterase (EC 3.1.4.17) from bovine ventricle was isolated and purified by (NH4)2SO4 precipitation and DEAE-cellulose and Affi-Gel Blue chromatography. The enzyme activity was enriched 800-fold by these procedures. 2. Sucrose-density gradient centrifugation, gel filtration and non-denaturing polyacrylamide-gel electrophoresis resolved a single enzyme species with an Mr of 89 000. 3. Sodium dodecyl sulphate/polyacrylamide-gel electrophoresis of the purified enzyme demonstrated a prominent protein band at Mr 59000 and a minor band of Mr 28000. Calmodulin was not detected. 4. The hydrolysis of micromolar concentrations of 3':5'-cyclic guanosine monophosphate (cyclic GMP) but not 3':5'-cyclic adenosine monophosphate (cyclic AMP) was stimulated by calmodulin. 5. Anomalous biphasic kinetics plots were observed for both the catalysis of cyclic AMP and cyclic GMP hydrolysis. Kinetic plots became linear in the presence of calmodulin. 6. After several months of storage at -20 degrees C, the 5.3-S enzyme was transformed into a 6.2-S cyclic GMP-specific enzyme and a 4.4-S non-specific form. PMID- 6291509 TI - Heterogeneity of the calcium-dependent phosphatidylinositol phosphodiesterase in rat brain. AB - 1. The Ca(2+)-dependent phosphatidylinositol phosphodiesterase (phospholipase C type) from the cytosolic supernatant of rat brain was active against exogenous [(32)P]-phosphatidylinositol from pH5.0 to pH8.5. However, the activity in the range pH7.0-8.5 could not be recovered after precipitation with (NH(4))(2)SO(4); most of the enzyme activity was recovered in the 30-50% fraction and showed a single sharp pH optimum at 5.5. 2. The cytosolic supernatant was analysed by isoelectric focusing on acrylamide gels, and assay at pH5.5. Four peaks of phosphodiesterase activity were found at pI ranges 7.4-7.2, 6.0-5.8, 4.8-4.4 and 4.2-3.8. 3. The cytosolic supernatant was also applied to a chromatofocusing column, and again assayed at pH5.5. Four peaks were eluted: minor, but consistent, activity at the beginning of the elution with a pI of near 7.2 or above; a second peak at pH6.0-5.85; a third broad peak with a wide range pH5.3 4.2; and a fourth peak, which was eluted by washing the column with 1m-NaCl, suggesting an isoenzyme with a pI below 4.0 (supported by the result of the isoelectric focusing). 4. If all the chromatofocusing fractions were assayed at pH7.0 or 8.0 (at 1mm-Ca(2+)), only a single sharp peak was detected, with a pI of 4.6-4.8. This peak disappeared on (NH(4))(2)SO(4) fractionation (30-50%) of the cytosolic supernatant, whereas the four peaks with activity at pH5.5 were virtually unaffected. 5. The four activities (assayed at pH5.5) separated by chromatofocusing produced inositol 1:2-cyclic monophosphate, inositol 1 monophosphate and diacylglycerol as enzymic products. 6. We conclude that the Ca(2+)-dependent phosphatidylinositol phosphodiesterase exhibits considerable heterogeneity, both with respect to pH optima of activity, and its isoelectric properties. PMID- 6291510 TI - Superoxide-dependent formation of hydroxyl radicals in the presence of iron salts is a feasible source of hydroxy radicals in vivo. PMID- 6291511 TI - Leukotriene B4 is a complete secretagogue in human neutrophils: a kinetic analysis. PMID- 6291512 TI - A critical evaluation of the possible modulation of hepatic microsomal glucose-6 phosphatase activity by protein phosphorylation. PMID- 6291513 TI - Cytosolic calcium dependent neutral proteinase of human erythrocytes: the role of calcium ions on the molecular and catalytic properties of the enzyme. PMID- 6291514 TI - The NH2- and COOH-terminal sequences of the angiotensin-converting enzyme isozymes from rabbit lung and testis. PMID- 6291516 TI - Cis-diamminedichloroplatinum(II) modified DNA stimulates far greater levels of S1 nuclease sensitive regions than does the modification produced by the trans- isomer. PMID- 6291515 TI - Inhibition of membrane phosphotyrosyl-protein phosphatase activity by vanadate. PMID- 6291517 TI - A purification of microsomal glucose-6-phosphatase from human tissue. PMID- 6291518 TI - Avidin induction by dibutyryl cyclic guanosine 3',5'-monophosphate in chick oviduct organ culture. PMID- 6291519 TI - Characterization of a cytochrome oxidase-deficient yeast mutant which accumulates porphyrins. PMID- 6291520 TI - The molybdenum and iron-sulphur centres of Escherichia coli nitrate reductase are non-randomly oriented in the membrane. PMID- 6291521 TI - Increased collagenase and gelatinase activities in keratoconus. PMID- 6291522 TI - Detergent-activation of latent collagenase and resolution of its component molecules. PMID- 6291523 TI - Prostaglandin A1 induces the synthesis of a new protein in cultured AGMK cells. PMID- 6291525 TI - Direction of zein gene transcription in maize genomic clones. PMID- 6291524 TI - Free radicals and tissue damage produced by exercise. PMID- 6291526 TI - Ubisemiquinone radicals in liver: implications for a mitochondrial Q cycle in vivo. PMID- 6291527 TI - 23-keto-25-hydroxyvitamin D3 and 23-keto-1,25-dihydroxyvitamin D3: two new vitamin D3 metabolites with high affinity for the 1,25-dihydroxyvitamin D3 receptor. PMID- 6291528 TI - Effects of estradiol administration in vivo on testosterone production in two populations of rat Leydig cells. PMID- 6291529 TI - Characterization of DNA from the rne gene of Escherichia coli: uniqueness of the rne DNA. PMID- 6291530 TI - Decrease of transferrin receptor during mouse myeloid leukemia (M1) cell differentiation. PMID- 6291531 TI - Ammonia detoxification in the fatty liver. PMID- 6291532 TI - The phosphorylation of intact erythrocytes by exogenously added cAMP-dependent protein kinase. PMID- 6291533 TI - Cyclic AMP-dependent phosphorylation of fructose-1,6-bisphosphatase in yeast. PMID- 6291534 TI - Activation of the branched-chain alpha-ketoacid dehydrogenase complex by a broad specificity protein phosphatase. PMID- 6291535 TI - Evidence that recycling of low density lipoprotein receptors does not depend on delivery of receptors to lysosomes. PMID- 6291536 TI - Identification of a protein component of the Ca2+-dependent K+ channel by affinity labelling with apamin. PMID- 6291537 TI - Acetylcholine receptor-controlled ion translocation. A comparison of the effects of suberyldicholine, carbamoylcholine, and acetylcholine. PMID- 6291538 TI - [3H]Dihydroergocryptine binding to alpha-adrenergic receptors of human platelets. A reassessment using the selective radioligands [3H]prazosin, [3H]yohimbine, and [3H]rauwolscine. AB - Which subtype(s) of the alpha-adrenergic receptor occurs on human platelets? Studies of platelet responsiveness to adrenergic compounds and indirect radioligand binding studies addressing this question have yielded contradictory conclusions. These bindings studies employed the ligand [3H]dihydroergocryptine ( [3H]DHE), an alpha-adrenergic antagonist that does not select between alpha 1- and alpha 2-adrenergic receptors and that also binds to other receptor types in some tissues. To determine the subtype of the platelet alpha-adrenergic receptor, we have examined the binding to intact human platelets of [3H]prazosin (alpha 1 selective), [3H]yohimbine (alpha 2-selective), and [3H]rauwolscine (alpha 2 selective), and we have compared the binding of these selective radioligands with that of [3H]DHE. [3H]Yohimbine and [3H]rauwolscine both bound with high affinity (Kd = 2.7 and 4.6 nM, respectively) to an equal number and a single class (Hill coefficient approximately 1.0) of sites ( approximately 300 per platelet), but [3H]yohimbine yielded lower nonspecific binding than did [3H]rauwolscine. In paired experiments, [3H]DHE bound to 1.5 times as many (phentolamine displaceable) sites as did [3H]yohimbine or [3H]rauwolscine. Unlabeled yohimbine and epinephrine competed for fewer [3H]DHE binding sites than did phentolamine. Thus, in addition to binding to the alpha 2-adrenergic receptors identified by [3H]yohimbine and [3H]rauwolscine, [3H]DHE seems to bind to other sites on human platelets. The nature of these sites is not clear. We found that [3H]prazosin did not identify alpha 1-adrenergic receptors on platelets, and that phenoxybenzamine only inhibited [3H]yohimbine and [3H]DHE binding at higher concentrations than usually observed for alpha 1-adrenergic receptors. We conclude that (1) all alpha adrenergic sites on human platelets are of the alpha 2 subtype, (2) [3H]DHE may bind to additional, as yet ill-defined, sites in addition to those sites identified by [3H]yohimbine and [3H]rauwolscine, and (3) [3H]yohimbine is the preferred antagonist radioligand for studying the alpha 2-adrenergic receptors on human platelets. PMID- 6291539 TI - The interaction of miconazole and ketoconazole with lipids. AB - Staphylococcus aureus can be protected by unsaturated unesterified fatty acids against the growth inhibitory effects of miconazole and ketoconazole observed at concentrations greater than 10(-6) M and greater than 10(-5) M, respectively. Miconazole's fungicidal activity is partly antagonized by oleic acid. However, the effect of ketoconazole on the viability of Candida albicans was not affected by this fatty acid. Cytochrome oxidase and ATPase activities are more sensitive to miconazole (10(-5) M) than to ketoconazole (greater than 10(-4) M) and also liposomes are more susceptible to lysis induced by miconazole. Using differential scanning calorimetry it is shown that high concentrations of miconazole shift the lipid transition temperature of multilamellar vesicles to lower values without affecting the enthalpy of melting. Ketoconazole induces a broadening of the main transition peak only. It is suggested that miconazole changes the lipid organization without binding to the lipids, whereas ketoconazole is localized in the multilayer without having an important direct effect on the lipid organization. The results indicate that miconazole, and to a lesser extent ketoconazole, at doses that can be reached by topical application only, interfere with a third target (the two others are ergosterol synthesis and fatty acid elongation plus desaturation). It is hypothesized that the induced change in lipid organization may play some role in miconazole's topical antibacterial and fungicidal activity, whereas it does not seem to play a significant role in ketoconazole's activities. PMID- 6291540 TI - Multiple effects of a new anti-inflammatory agent, timegadine, on arachidonic acid release and metabolism in neutrophils and platelets. AB - Casein-elicited rat peritoneal polymorphonuclear leukocytes (PMNL) and rabbit platelets were prelabelled with [1-14C]arachidonic acid, and the effect of timegadine, a new anti-inflammatory agent, on the release and metabolism of arachidonic acid induced by A23187 (PMNL) and thrombin (platelets) was studied and compared with the effect of other compounds reported to affect these enzymatic mechanisms. Timegadine inhibited arachidonic acid release from both cells (IC50 = 2.7 X 10(-5) M), the lipoxygenase activity in PMNL (IC50 = 4.1 X 10(-5) M) and the cyclooxygenase activity in platelets (IC50 = 3.1 X 10(-8) M). By these mechanisms, PMNL leukotriene B4 formation was inhibited by 50% at 2.0 X 10(-5) M, platelet thromboxane B2 at 3.2 X 10(-8) M, and platelet 12-HETE at 4.9 X 10(-5) M. These effects might add to the understanding of the anti-inflammatory properties of timegadine. PMID- 6291541 TI - Direct action of D-lysergic acid diethylamide on dispersed mucosal cells from guinea pig stomach. AB - In dispersed mucosal cells from guinea pig stomach, D-lysergic acid diethylamide (LSD) was a partial agonist with respect to histamine. LSD, like histamine, inhibited [3H]histamine binding and increased both cellular cyclic AMP and [14C]aminopyrine uptake by interacting with histamine H2-receptors on parietal cells. These processes were blocked by both histamine H1- and H2-antagonists and, thus, provide evidence that histamine H2-receptors on guinea pig parietal cells resemble those in brain tissue in that they interact with LSD as well as with both classes of histamine antagonists. PMID- 6291542 TI - Interactions of agonists and antagonists with a novel type of GABA receptor. PMID- 6291543 TI - Protective effect of diethyldithiocarbamate and carbon disulfide against liver injury induced by various hepatotoxic agents. AB - Diethyldithiocarbamate (DTC) and carbon disulfide (CS2), at nearly equimolar oral dose levels, protected mice against liver damage induced by carbon tetrachloride, chloroform, bromotrichloromethane, thioacetamide, bromobenzene, furosemide, acetaminophen, dimethylnitrosamine and trichloroethylene, as evidenced by the suppression of elevations in plasma GPT activity and liver calcium content, and of histopathological alterations. Both agents also prolonged hexobarbital sleeping time and zoxazolamine paralysis time in mice. DTC and SC, alone, given orally, decreased microsomal metabolism of several substrates (aniline, p nitroanisole, hexobarbital, zoxazolamine, aminopyrine and 3,4-benzopyrene), CC14 induced lipid peroxidation, and cytochrome P-450 content. The loss of microsomal drug-metabolizing enzyme activity was also observed in the experiments in vitro using liver slices and isolated microsomes. Since a characteristic common to such diverse hepatotoxins is that they require metabolic activation before exhibiting hepatotoxicity, the protective mechanisms of DTC and CS2 may involve their interference with the process of metabolic activation of these hepatotoxins. The protective action of DTC may be mediated almost entirely through CS2 when administered orally and at least partly with parenteral administration, since, in CCl4-induced liver injury, DTC was most effective when given orally, while the action of CS2 was less dependent on the route of administration. Thus CS2 and CS2 producing agents in vivo such as dithiocarbamate derivatives and disulfiram may modify toxicological and pharmacological effects of foreign compounds by inhibiting microsomal drug-metabolizing enzyme activity in the liver. PMID- 6291544 TI - Selective inhibition of separated forms of cyclic nucleotide phosphodiesterase from rat heart by some pentasubstituted quercetin analogs. AB - Synthetic analogs of quercetin were evaluated as inhibitors of cyclic nucleotide phosphodiesterase in rat heart preparations. Three main enzymatic forms of cyclic nucleotide phosphodiesterase can be resolved from rat heart cytosol by isoelectric focusing. Inhibition studies were performed with the whole cytosolic and particulate preparations and with the cytosolic separated enzymatic forms. All the compounds studied proved more potent as inhibitors of the particular preparation than as inhibitors of the cytosolic fraction. With the exception of water-soluble derivatives and quercetin, they showed a better potency to inhibit cyclic AMP than cyclic GMP phosphodiesterase activity of the cytosolic and particulate preparations; likewise, among the three separated enzymatic forms, the cyclic AMP specific form of pI 5.55-6 is the most inhibited by these flavonoid compounds. In all cases, the highest selectivity was observed with pentaethyl quercetin (2). PMID- 6291545 TI - The role of the superoxide and hydroxyl radicals in the degradation of DNA and deoxyribose induced by a copper-phenanthroline complex. AB - DNA degradation by a copper(II)-phenanthroline complex was studied in the presence of NADH, 2-mercaptoethanol or a mixture of hypoxanthine and xanthine oxidase, which generates the superoxide radical, O2-. In all cases degradation was prevented by catalase but not by scavengers of the hydroxyl radical, OH. It remains possible, however, that OH was generated in close association with DNA so that the scavengers could not remove it before it reacted. Superoxide dismutase inhibited DNA degradation at low copper (II) phenanthroline concentrations in the presence of NADH or hypoxanthine-xanthine oxidase, but not at higher complex concentrations. Superoxide dismutase had little effect on DNA degradation in the presence of 2-mercaptoethanol. The role of oxygen radicals in the DNA degradation induced by copper(II) phenanthroline is discussed. PMID- 6291546 TI - Subunit structure of rat liver alpha 1 adrenergic receptor. PMID- 6291547 TI - Prolactin: multiple intracellular processing routes plus several potential mechanisms for regulation. PMID- 6291548 TI - Photoaffinity labelling of beta-adrenergic receptors of C6 glioma cells. Presence of a nucleophilic group in the receptor. PMID- 6291549 TI - Conjugation of dopa and 5-S-cysteinyldopa with cysteine mediated by superoxide radical. AB - cytotoxicity of catechols has been ascribed to their binding with proteins through sulfhydryl groups. Superoxide radical (O2-) generated in hypoxanthine xanthine oxidase system at pH 7.4 mediated conjugation of dopa with cysteine to form cysteinyldopas. Similarly, 5-S-cysteinyldopa gave 2,5-S,S-dicysteinyldopa. The rates of oxidation of the catechols by O2- appear to be comparable to that of reduction of nitro-blue tetrazolium by O2-. These results suggest that catechols may exert cytotoxicity in cells where biochemical defence against O2- or the quinone oxidation products is not sufficient. PMID- 6291550 TI - Differential effects of sedative and anticonvulsant barbiturates on specific [3H]GABA binding to membrane preparations from rat brain cortex. AB - The sensitivity to barbiturates of [3H]GABA binding to synaptosomal membrane fractions from rat cortex has been examined. We show that a range of anaesthetic/sedative barbiturates enhance GABA binding in the presence of chloride or other ions that interact with the associated ionophore. Furthermore, picrotoxinin and the anticonvulsant barbiturate phenobarbital antagonise the enhancement produced by pentobarbital. These effects are therefore comparable to those observed at benzodiazepine receptors and may be mediated through the chloride ionophore component of the receptor complex. Other classes of anticonvulsants failed to antagonise pentobarbital activation, suggesting that these interactions may occur at a specific barbiturate site in the membrane. PMID- 6291551 TI - Alteration of sodium and potassium mobilization and of adrenal function by long term ingestion of lead. AB - Serum sodium concentration was markedly decreased by long-term (12 weeks) ingestion of lead above 5 mg Pb . kg-1 . day-1, whereas serum potassium concentration was notably decreased by the long-term (12 weeks) ingestion of lead above 2 mg Pb . kg-1. Urinary sodium and potassium in fasted rats were increased markedly 24 hr after a single lead dose (200 mg Pb/kg, o.p.) [Y. Suketa, S. Hasegawa and T. Yamamoto, Toxic. appl. Pharmac. 47, 203 (1979)]. In contrast, urinary excretion of sodium or potassium in non-fasted rats was not changed significantly by 2 weeks of lead ingestion at 200 mg Pb . kg-1 . day-1. Renal activities of Na+, K+-ATPase and K+-dependent phosphatase were decreased to 50 70% of control values by long-term (12 weeks) ingestion of lead (above 5 mg Pb . kg-1 . day-1). PMID- 6291552 TI - Interaction of psychotropic drugs with phospholipids. AB - The interactions of psychotropic drugs with phospholipids, including lysophosphatidylcholine, phosphatidylinositol, and lysophosphatidylserine, were studied using calmodulin-dependent cyclic nucleotide phosphodiesterase partially purified from the cortex of hog brain. All the compounds used inhibited both calmodulin- and phospholipid-stimulated phosphodiesterase activity but not the basal activity. Fluphenazine was confirmed by kinetic analysis to be a competitive inhibitor, with both calmodulin and phospholipid. Using fluphenazine Sepharose affinity chromatography, it was demonstrated that fluphenazine did not interact with the enzyme. The potencies of antipsychotics such as fluphenazine in inhibiting the various phospholipid-dependent activations decreased in the following order: lysophosphatidylcholine-, phosphatidylinositol-, and lysophosphatidylserine-dependent activation. On the other hand, antidepressant drugs exhibited similar inhibitory potencies towards lysophosphatidylcholine- and phosphatidylinositol-dependent activation. Antipsychotic and antidepressant drugs appear to have different characteristics with regard to lipid-drug interaction. PMID- 6291553 TI - Polyarthritis with birefringent lipid within synovial fluid macrophages: case report and ultrastructural study. PMID- 6291554 TI - [Decreased sodium conductance of the cardiomyocyte membrane: the cause of the negative inotropic action of quinidine-like anti-arrhythmia agents]. AB - The action of tetrodotoxin on the guinea-pig muscle caused a decrease in the maximum rate of action potential, which was accompanied by a negative inotropic effect. The magnitude of such an effect correlated well with the decrease of sodium permeability, measured by the maximum rate of depolarization (r = 0,995). The effect was enhanced under the action of veratrine and diminished under that of isoproterenol. It was suggested that the negative inotropic effect of tetrodotoxin was the result of the inhibition on Na-Ca exchange due to diminished intracellular Na+ concentration. PMID- 6291555 TI - [Plasma insulin and glucagon during experimental MHV-3 virus hepatitis in mice]. AB - Insulin and glucagon values were determined in the plasma of mice during MHV-3 experimental viral infection. The results showed a different behaviour of the two hormones. The insulin values remained normal until the 12th hour of infection, with a statistically significant increase as from the 12th hour up to their maximum peak at the 24th hour, remaining constant at that level until the 72nd hour. As for the glucagon values there was a statistically significant decrease until the 24th hour of infection when they suddenly began to increase up to their maximum peak at the 48th hour remaining constant at that level until the 72nd hour. Increased of the insulin values may be attributed to liver cell damage because of the decreased metabolization of the hormone. As for glucagon the results of the decreased values during the initial phase of the infection may be the concentration decrease of free plasma amino acids, whereas the increased values during the subsequent phases is probably caused by the concentration increase of free plasma amino acids, of hypogycemia and of stress. PMID- 6291556 TI - [Biochemical and ultrastructural research on the antagonism between vitamins A and D at the leve of the erythrocyte membrane]. AB - The reason for the antagonistic action between retinol and cholecalciferol on cellular membranes was investigated "in vitro" on human erythrocytes, by testing their lipoperoxide production, the amount of membrane-bound retinol and the morphology of these cells by scanning electron microscope. Lipoperoxides were assayed by TBA-test; the membrane-bound retinol was tested by the Karr-Price method on lipids extracted from red blood cells by the Folch method and vitamin D deprived by TCL chromatography; the morphological changes of these cells treated with vitamin A, or D, or A+D, were tested by scanning electron microscope. In our experimental system both retinol and cholecalciferol increase the lipoperoxide production from the erythrocytes; nevertheless no synergism of action can be demonstrated between the two vitamins. Cholecalciferol decreases the amount of membrane-bo und retinol as well as the changes produced by vitamin A in red blood cell surface. These results are in good agreement with our previous observations obtained by nuclear magnetic resonance, using the same experimental system. PMID- 6291557 TI - Antioxidant activity of bull semen in relation with aging. AB - This paper reports on the findings of an antioxidant activity in whole semen from bull and its components, washed spermatozoa and seminal fluid. The antioxidant activity has been evaluated as the ability of semen or its components to inhibit the spontaneous autooxidation of epinephrine a pH 10, 2, which involves the production of superoxide radicals (0(2)). This preliminary study provides further evidence on the role of free radicals, which are well known to be dangerous to cellular life, and points to the need of a better understanding of the role of antioxidant activities as a cellular protective mechanism. PMID- 6291558 TI - [Isolation and characterization of epithelial cells of colon mucosa: preliminary results]. AB - A procedure for the isolation of epithelial cells from rat and human large bowel is described; the minced tissue was carefully washed with saline and incubated for 45 min in a collagenase-jaluronidase solution; the dispersion of the epithelial cells was achieved by a subsequent treatment with the calcium chelator EDTA. The isolated cells were characterized by cytological and histochemical (PAS, alkaline phosphatase, N-acetyl-D-glucosaminidase) procedures; viability index was assessed by the trypan blue exclusion test; the ability to grow in culture on both liquid and semisolid media was also tested. This method can be successfully applied either to normal or neoplastic colonic mucosa, the resulting cell suspension being suitable for further characterization. PMID- 6291560 TI - Inhibitor effect of polyamines on reduction of cytochrome C by superoxide anion. AB - The interaction of polyamines with enzymatically generated free radicals was investigated. The superoxide anion (O-2) was generated in vitro using the xanthine oxidase-hypoxanthine system. Our results show that spermidine or spermine at different concentrations (20-200 mM) inhibit the reduction of cytochrome c; the highest levels of inhibition were obtained adding 200 mM spermidine or spermine. Putrescine (200 mM) affected the reduction of cytochrome c very little. PMID- 6291559 TI - Functional impairment of intact rat liver cells due to biological aldehydes. AB - The addition of lipid peroxidation end-products, 4-hydroxynonenal (HNE) or hexanal (HEX) to the incubation medium of rat hepatocytes caused significant decrease of cell cytochrome P-450 content and inactivation of total cell glucose 6-phosphatase. Both the tested aldehydes exerted a marked inhibition of triglyceride secretion by liver cells. The reported results on intact cells furtherly support a possible damaging effect of aldehydes in pathological conditions in which a stimulation of lipid peroxidation occurs. PMID- 6291561 TI - [Schmidt Ruppin-D-ASV-induced primary rat brain tumor model for therapeutic screening]. AB - It is important to evaluate the therapeutic and side effects of new therapy for malignant brain tumors in an adequate animal model prior to its initial clinical investigation. For decades, neurooncologists have argued for the use of primary, autochthonous tumors rather than transplanted tumors such as C 6 glioma cells and 9 L gliosarcoma cells. But unfortunately, no spontaneous animal astrocytomas are currently available as usable models. So we tried to establish the model of primary, autochthonous avian sarcoma virus-induced rat gliomas for experimental chemotherapy and immunotherapy. The present study was undertaken to determine the incidence and histologic pattern of tumors and the mean survival time of the animal model used. It was found that the intracerebral inoculation of 2 X 10(6) FFU/5 microliter of infectious cell free homogeneous subgroup D Schmidt-Ruppin avian sarcoma virus (SR-D-ASV) into 3-day-old inbred Fischer 344 rats induced small sized tumors in all rats 20 days later. The mean survival time of inoculated rats were 58.7 +/- 12 days. As to the classification of SR-D-ASV induced brain tumors in Fischer rats, astrocytoma was 70.6% (protoplasmic astrocytoma 23.5%, fibrillary astrocytoma 47.1%), sarcoma 17.6%, and mixed astrocytoma and sarcoma 11.8%. In conclusion, this SR-D-ASV induced tumor in the rat fulfilled the following criteria for the desirable animal model: (1) Spontaneously arising. (2) Glial origin. (3) Intraparenchymal growth. (4) Uniformly fatal within reasonable time period. Statistic evaluation of the effects of chemotherapy and immunotherapy was considered to be possible. PMID- 6291562 TI - [Receptors in positron emission tomography]. PMID- 6291563 TI - Long-term efficacy of angiotensin-converting-enzyme inhibition with captopril in mild-to-moderate essential hypertension. AB - 1 Thirty-one patients with mild-to-moderate essential hypertension were treated with captopril for 30 months. 2 Captopril effectively lowered raised blood pressure in hypertensive patients with high, low, and normal renin concentrations. There was no significant change in heart rates. Captopril alone normalised blood pressure (diastolic pressure below 95 mm Hg) in 11 patients. In the other patients the addition of hydrochlorothiazide produced a further hypotensive effect. Blood pressure control could be maintained without any signs of tachyphylaxis. 3 Plasma concentrations of aldosterone and angiotensin II were significantly lower after 30 months of therapy than pretreatment values whether captopril was associated with diuretics or not. No relevant symptomatic or biochemical adverse reactions were observed. 4 These data establish the long-term potential of captopril for outpatient therapy in mild-to-moderate forms of essential hypertension. PMID- 6291565 TI - The effect of misonidazole combined with WR2721 on tumour response and leucopenia due to cyclophosphamide or melphalan. PMID- 6291564 TI - Sustained haemodynamic and clinical effects of captopril in long-term treatment of severe chronic congestive heart failure. AB - 1 The angiotensin-converting-enzyme inhibitor captopril is known to produce beneficial haemodynamic effects in patients with chronic congestive heart failure. 2 Twelve patients with chronic congestive heart failure were conventionally treated with digitalis and diuretic therapy plus oral captopril (75-150 mg/day), and 14 patients were used as a control group. 3 There was no improvement in functional and haemodynamic values in the controls, but the patients treated with captopril showed a significant functional improvement (increase of exercise time and improvement in New York Heart Association functional class), a definitive decrease in systemic vascular resistance (20%), and a significant increase in the cardiac index and ejection fraction (18% and 28% respectively). PMID- 6291566 TI - Epidermal surface receptors which link pharmacological mediators to the adenylate cyclase system. AB - The major purpose of our studies has been to investigate various stimulators of the epidermal adenylate cyclase system. We have recognized the occurrence of four distinct adenylate cyclase systems which respond respectively to catecholamine, histamine, prostaglandin and adenosine. The exposure of floating skin slices in vitro to a stimulator causes a rapid intracellular accumulation of cyclic AMP, which is always transient. Further addition of the same stimulator will not stimulate the same receptor system against the state of 'refractoriness'. The addition of any of the other stimulators can increase the cyclic AMP level. Furthermore, the fact that each stimulator can yield an 'additive' stimulatory effects leads to the conclusion that the epidermis has four distinctly specific and independent adenylate cyclase systems. Our recent investigations have been directed to the analyses of subunits of these skin surface receptor-adenylate cyclase systems. We used two experimental systems, i.e. one being a 'leaky' cell system in which its subunits such as receptor, GTP-regulatory protein and the catalytic unit (adenylate cyclase) are still linked together, and the other being independent preparations of the receptor and catalytic units (with GTP-regulatory protein). These systems allowed us to probe the cell membrane from the inside as well as from the outside. Some of the preliminary kinetic data are herein introduced. PMID- 6291567 TI - Effect of beta-adrenergic receptor blockade or refractoriness induced by isoproterenol on growth of keratinocytes in vitro. AB - Almost complete suppression of beta-adrenergic sensitivity of adenylate cyclase was produced in keratinocytes growing on collagen gels either by induction of refractoriness by isoproterenol or by receptor blockade by propranolol, these drugs being introduced 3 days post-plating and continuously thereafter. Neither of these two manoeuvres affected growth of the keratinocytes. Despite this suppression, basal levels of cyclic AMP never fall below the levels observed in untreated cultures. On the other hand, growth inhibition is observed if isoproterenol or propranolol is added on plating. When keratinocytes are seeded on 3T3 fibroblast feeder layers, isoproterenol enhances colony growth, as reported elsewhere, indicating that the effects of isoproterenol are therefore modulated by the culture surface. In either of these in vitro model systems studied (collagen or feeder layers), no mechanism was obvious that would account for propranolol-induced psoriasiform lesions observed in vivo. PMID- 6291568 TI - High affinity membrane receptors in cultured human keratinocytes. I. The beta adrenergic receptors. PMID- 6291569 TI - Kinetic studies on MSH and alpha 2-adrenoceptor interaction on the Anolis melanophore. PMID- 6291570 TI - Adrenoceptor-agonist inhibition of the histamine-induced cutaneous response in man. PMID- 6291571 TI - Glucocorticoid action: a mechanism involving nuclear and non-nuclear pathways. PMID- 6291572 TI - Multiple steroid binding sites in human skin cytosol. PMID- 6291573 TI - Steroid hormone receptor analysis in human melanoma and non-malignant human skin. PMID- 6291574 TI - Lymphocyte receptors. PMID- 6291575 TI - The visualization of plasma membrane receptors. PMID- 6291576 TI - Mitochondrial NADH dehydrogenase in iron-deficient and iron-repleted rat muscle: an EPR and work performance study. AB - Iron may affect both respiratory O2 transport and mitochondrial electron transport in the performance of muscle work. This study was designed to elucidate the molecular defect of iron-deficient work performance by identifying heretofore unmeasurable mitochondrial enzymes that are diminished by iron deficiency and may be restored by iron repletion. Female rats were made iron-deficient by dietary control and were repleted by oral iron. Iron deficiency reduced physical work capacity (treadmill running time), haemoglobin (Hb), and mitochondrial iron sulphur (Fe-S) centres in heart and skeletal muscles; mitochondrial number was unaffected. Oral iron supplementation restored work capacity and Hb within 4 d to normal or near-normal levels, but in general Fe-S centres of mitochondria due to NADH dehydrogenase remained at iron-deficient levels. Subnormal concentrations of mitochondrial iron-dependent NADH dehydrogenase in muscle are not by themselves rate-limiting in work performance. PMID- 6291577 TI - Philadelphia chromosome-positive blastic leukaemia: ultrastructural and ultracytochemical evidence of basophil and mast cell differentiation. AB - Ultrastructural, ultracytochemical, immunologic and biochemical studies were performed on leukaemic cells from 41 patients with Philadelphia chromosome positive blastic leukaemia; 28 patients were in blast transformation of chronic myelogenous leukaemia and 13 patients presented with 'acute' leukaemia. The patients were divided into two morphologic groups, lymphoid (16 cases) and myeloid (25 cases), on the basis of light microscopy and cytochemistry. All lymphoid cases studied for the presence of CALLA (10 patients) and TdT (11 patients) were positive. Two of 13 myeloid cases studied were TdT positive. The blasts from 10 of 16 lymphoid cases contained immature basophil/mast cell granules on ultrastructural examination. Peroxidase-positive 'lymphoid' blasts were noted in three of seven patients studied by ultracytochemical techniques. The reactivity was primarily confined to granular structures. Of the 25 cases in the myeloid group, blasts from 14 cases showed basophil/mast cell differentiation, nine cases showed neutrophil/monocyte features, and two cases were megakaryoblastic. Distinct patterns of ultrastructural peroxidase positivity were seen in the seven myeloid cases studied. In basophil/mast cell precursors the reactivity was primarily confined to granules; neutrophil precursors showed reactivity in the nuclear envelope, rough endoplasmic reticulum (RER), golgi and granules; in megakaryoblasts, only the nuclear envelope and RER were positive while the granules were consistently negative. PMID- 6291578 TI - Census-based mortality study of fertiliser manufactures. AB - This study was designed to investigate whether exposure to nitrate-containing dust during fertiliser manufacture was associated with an excess of deaths from cancer in general or specifically from cancers of the digestive tract, liver, lung, and bladder. It was based on data extracted from census schedules by the Office of Population Censuses and Surveys, occupational characteristics recorded by fertiliser workers at the 1961 and 1971 censuses of England and Wales being related to subsequent mortality ascertained through the National Health Service Central Register. The 1961 cohort, followed up until 1978, showed a "healthy worker effect" and no evidence of excess mortality from cancer at any site. The 1971 cohort also showed below average mortality during 1971-7 for all causes of death and for circulatory diseases, but there were more deaths from cancer than expected, due mainly to an excess of cancers of the lung and digestive tract. The excess of cancer was more pronounced, but not statistically significant, when compared with other employed men. Though the numbers for comparison were small, there was weak evidence of an association between cancer mortality and frequency of exposure to nitrate-containing dust in this cohort. It is difficult to reconcile the excess cancer mortality in the 1971 cohort with the more favorable level in the earlier cohort, since industrial hygiene has improved and the cohorts showed a similar distribution by region and social class. To examine further these conflicting results the 1971 cohort will be followed for a longer period and re-examined when more deaths have accrued. PMID- 6291579 TI - Lung function and radiographic change in chrysotile workers in Swaziland. AB - The effect on lung function and radiographic indices of exposure to chrysotile asbestos was investigated by cross-sectional studies in two groups of men at Havelock Mine, Swaziland. The first group consisted of 214 employees and ex employees, mean age 52, who had been employed for at least 10 years, and whose dust exposure ranged from minimal for surface workers to very heavy for those in the grading and bagging sections of the mill. In this group 29% had category 1 or more simple pneumoconiosis and 4.5% category 2 or more. For surface and mine workers, the estimated annual deterioration in FEV1 and FVC and the increase in category of pneumoconiosis was similar to that due to age alone, while the heaviest exposure almost doubled the decline in lung function and trebled the rate of progression of pneumoconiosis. The second group consisted of 224 men, mean age 33, all currently working in the mill and having been employed there for at least a year. In this group 30% had category 1 or more simple pneumoconiosis, and 2.7% category 2. Exposure in the dustiest sections of the mill more than doubled the estimated annual decline in lung function and doubled the rate of progression of pneumoconiosis. PMID- 6291580 TI - Mortality of two groups of women who manufactured gas masks from chrysotile and crocidolite asbestos: a 40-year follow-up. PMID- 6291582 TI - Cholesterol-phospholipid interaction in membranes. 1. Cholestane spin-label studies of phase behavior of cholesterol-phospholipid liposomes. AB - The effect of cholesterol concentration on the thermotropic phase behavior of aqueous phospholipid multi-bilayers was monitored by means of electron spin resonance spectroscopy (ESR) of a cholestane spin-label (CSL). The spin-label itself induces an additional transition in several different phospholipids, which is attributed to local melting around the spin probe. In contrast, cholesterol prevents its neighboring phospholipids from undergoing fluidization. Small additions of cholesterol affect the position of the probe-induced lipid mobilization curve. The phospholipid main gel-liquid-crystal transition, which is also observed as a separate change in probe mobilization, is not affected by low concentrations of cholesterol. These observations indicate the presence of two phases, a cholesterol-rich phase and a pure phospholipid phase, and indicate that CSL preferentially enters the cholesterol-rich phase. Addition of more than 20 mol % cholesterol abolishes the bulk phospholipid phase. This is evidenced by the disappearance of the gel-liquid-crystal transition as observed by ESR. However, the CSL-induced transition is present at all concentrations of cholesterol and CSL. The behavioral differences between the two sterols caution against using this probe as a direct substitute for cholesterol. However, it remains a useful tool for monitoring the phase behavior of cholesterol-phospholipid bilayer systems. PMID- 6291581 TI - Activation of spin-labeled chicken pepsinogen. AB - Chicken pepsinogen has been spin-labeled by the attachment of four nitroxides to epsilon-amino groups near the protein's amino terminus. Acidification results in a bond cleavage, generating a nonlabeled, enzymatically active protein. Electron spin resonance spectra of the spin-labeled zymogen, acidified in the presence or absence of pepstatin, are identical and indicate that the nitroxides are quite mobile, compared to the nonacidified zymogen. This mobilization is interpreted as the freeing of the peptide to which the spin-labels are attached, from the protein, subsequent to the acidification that causes a peptide bond cleavage. The rate at which the peptide leaves the protein is 1 order of magnitude slower than the cleavage of the peptide bond, measured by the rate of appearance of milk clotting activity (first-order rate constants of 0.3 min-1 vs. 6 min-1 at pH 2, 22 degrees C). The inclusion of pepstatin, at molar ratios above 2 during activation, decreases the rate of peptide leaving. These observations, and those previously reported for activation of spin-labeled pig pepsinogen, are incorporated into a model of pepsinogen activation. PMID- 6291583 TI - Intramolecular electron transfer in Chlorobium thiosulfatophilum flavocytochrome c. AB - The electron-transfer reactions of photoproduced lumiflavin semiquinone and fully reduced lumiflavin with oxidized Chlorobium thiosulfatophilum flavocytochrome c have been studied by using laser flash photolysis. The Chlorobium flavocytochrome c contains one heme and one flavin per Mr 50 000, and thus the possibility exists for intramolecular electron transfer. We find a complex kinetic pattern which is consistent with the transient formation of a spectrally perturbed protein-bound flavin semiquinone which transfers an electron intramolecularly to the heme (k = 1 X 10(3)-1.8 X 10(3) s-1 for the neutral semiquinone, depending upon the pH). Evidence is presented that electron transfer from exogenous lumiflavin to the heme moiety occurs through the protein-bound flavin. We have also performed redox titrations which determine the midpoint potentials of the heme and flavin prosthetic groups at various pH values and the pK values for the semiquinone (6.4) and fully reduced flavin (6.1). Thus, at pH 7, the semiquinone is predominantly in the anionic form at equilibrium. The reactions of Chlorobium flavocytochrome c with photoreduced lumiflavin are similar to those previously found with Chromatium vinosum flavocytochrome c [Cusanovich, M. A., & Tollin, G. (1981) Biochemistry 19, 3343-3347] in that a protein-bound flavin semiquinone is an intermediate in the pathway of reduction. However, the rate constants are substantially different. As a class, the flavocytochromes c appear to operate by analogous mechanisms involving rapid intramolecular transfer between the heme and flavin moieties. PMID- 6291584 TI - Cytochrome c is cross-linked to subunit II of cytochrome c oxidase by a water soluble carbodiimide. AB - Modification of beef heart cytochrome c oxidase with 1-ethyl-3-[3 (dimethylamino)propyl]carbodiimide (EDC) or 1-ethyl-3-[3 (trimethylamino)propyl]carbodiimide (CH3EDC) has been found to significantly inhibit the high-affinity phase of the reaction of this enzyme with cytochrome c. Reaction conditions leading to a 70% inhibition of Vmax resulted in a 16-fold increase in the Km for cytochrome c. The loss in activity was accompanied by modification of subunit II to form a new species, II', which migrated somewhat more rapidly than the unmodified subunit during sodium dodecyl sulfate (NaDodSO4) gel electrophoresis. This new species was the major site of radiolabeling when cytochrome c oxidase was treated with [14C]CH3EDC, indicating covalent incorporation of the carbodiimide. Equimolar concentrations of cytochrome c dramatically protected cytochrome c oxidase from inhibition by the carbodiimide and in approximately the same proportion shielded subunit II from modification to the labeled II' species. In addition, cytochrome c was cross-linked to subunit II to form a new species migrating somewhat faster than subunit I during NaDodSO4 gel electrophoresis. This cross-linked species was shown to contain subunit II by using subunit-specific antibodies. We propose that EDC or CH3EDC reacts with one or more partially buried carboxyl groups on subunit II to form a positively charged N-acylurea which inhibits cytochrome c binding. In the presence of cytochrome c, EDC promotes formation of amide cross-links between lysine amino groups on cytochrome c and their complementary carboxyl groups on cytochrome c oxidase. PMID- 6291585 TI - Structure of the Glycyl-L-histidyl-L-lysine--copper(II) complex in solution. AB - Optical, electron paramagnetic resonance, and electron spin-echo envelope spectroscopies were used to examine the structure of the Cu(II) complex of glycyl L-histidyl-L-lysine (GHL) in solution. At neutral pH, GHL forms a mononuclear 1:1 Cu(II) compound having an EPR spectrum resembling that of Cu(II) equatorially coordinated by two or three nitrogen atoms. Electron spin-echo studies demonstrate that one of these is located in the histidyl imidazole ring. A pH titration of Cu(II)-GHL shows three optical transitions with apparent pKs of 3.6, 9.2 and 11.4 and molecularities, with respect to protons, of 2, 2, and 1, respectively. At the lowest pK, GHL binds Cu(II), forming the species present at physiological pH. At elevated pH, spectroscopic experiments suggest that an alteration of the Cu(II) structure occurs, yet the bound imidazole is retained. These solution studies are consistent with nitrogen coordination of Cu(II) in Cu(II)-GHL, but the solid-state polymeric structure, with oxygen-bridged Cu(II) pairs as previously determined by X-ray crystallographic analysis [Pickart, L., Freedman, J. H., Loker, W. J., Peisach, J., Perkins, C. M., Steinkamp, R. E., & Weinstein, B. (1980) Nature (London) 288, 715-717; C. M. Perkins, N. J. Rose, R. E. Steinkamp, L. H. Jensen, B. Weinstein, and L. Pickart, unpublished results], does not exist in solution. PMID- 6291586 TI - Orientation and role of nucleosidediphosphatase and 5'-nucleotidase in Golgi vesicles from rat liver. AB - The fate of UDP formed during the galactosylation of added N-acetylglucosamine in Golgi vesicles isolated from rat liver using D2O-sucrose gradients has been determined. UDP-Gal labeled with [14C]uracil was used, and the products of the reaction were separated and quantitated by using high-pressure liquid chromatography. [14C]Uridine rather than [14C]UDP or [14C]UMP was found to accumulate, indicating the presence of both UDPase and UMPase activities in the Golgi. Golgi vesicles were shown to contain a nucleosidediphosphatase activity that is membrane bound. It appears to be located on the luminal face of the Golgi since it is activated 3-5-fold by detergents and 4-fold by treatment of the vesicles with Filipin. We have shown previously that Filipin disrupts the Golgi but does not solubilize membrane-bound enzymes. The nucleosidediphosphatase of the Golgi differs from that present in rough endoplasmic reticulum in its absolute requirement for Ca2+ for activity and in its substrate specificity that is higher for UDP than for IDP. Golgi vesicles also contain UMPase activity that is stimulated only 2-fold by detergents or Filipin. Concanavalin A inhibits this activity about 80% in both intact and detergent-treated vesicles. The Golgi UMPase is thus probably identical with 5'-nucleotidase. These results are consistent with histochemical evidence from other laboratories that indicate that 5'-nucleotidase is present on both sides of liver Golgi membranes. In the presence of concanavalin A and N-acetylglucosamine, intact Golgi vesicles were found to convert UDP-Gal to UMP. These findings indicate that UDP formed by galactosyltransferase in the lumen of the vesicles is rapidly converted to UMP by UDPase in the lumen but that UMP moves rapidly out of the lumen of the Golgi and is broken down to uridine by 5'-nucleotidase on the cytoplasmic side of the vesicles. PMID- 6291587 TI - Interaction of the HpaI endonuclease with synthetic oligonucleotides. AB - To determine which functional groups of bases within the grooves of double helical DNA interact with the HpaI endonuclease, we have employed chemically synthesized octanucleotides containing base analogues. The 5-methyl group of thymine was probed as a contact between the HpaI endonuclease and its recognition sequence by using the oligonucleotides d(G-G-T-T-A-A-C-C), d(G-G-T-U-A-A-C-C), and d(G-G-T-U(Br)-A-A-C-C). The 2-amino group of guanine was probed as a contact for the HpaI endonuclease by using the octanucleotide d(G-I-T-T-A-A-C-C). The HpaI endonuclease cleaves octanucleotides d(G-G-T-T-A-A-C-C) and d(G-G-T-B-A-A-C C) according to Michaelis-Menten kinetics. However, both the Km and turnover number for d(G-G-T-B-A-A-C-C) were severalfold lower than those for cleavage of d(G-G-T-T-A-A-C-C). In addition, d(G-G-T-U-A-A-C-C) was not cleaved by HpaI endonuclease, suggesting that the 5-methyl group of thymine is a contact between the HpaI endonuclease and its recognition sequence. d(G-I-T-T-A-A-C-C) was not cleaved by the HpaI endonuclease which may be due in part to the low thermal stability of the duplex. Nevertheless, our results suggest that the 2-amino group of guanine is a contact for the HpaI endonuclease. A phosphate group 5' external to the HpaI recognition sequence has been identified as a contact between the HpaI endonuclease and DNA. The HpaI endonuclease cleaved 5'-phosphorylated octanucleotide 30-fold faster than unphosphorylated octanucleotide. In addition, the Km of the d(G-G-T-T-A-A-C-C) was 8000-fold higher than the Km of the phage f1 RFI DNA, suggesting that the octanucleotide is too short to take advantage of the entire DNA binding site of the enzyme. PMID- 6291588 TI - Comparison of transfer ribonucleic acid structures using cobra venom and S1 endonucleases. AB - Cobra venom nuclease V1, which cleaves double-stranded RNA, has been used to study the structure of four Escherichia coli tRNAs: Phe, Glu2, Leu2, and Ile1. The cleavage patterns are compared to those found for yeast tRNAPhe, the three dimensional structure of which is known. The cleavage patterns of all the tRNA molecules are similar, and the most sensitive cleavage is found at the central base pair of the anticodon stem. Studies of E. coli tRNALeu2, which has a large variable loop, are consistent with the formation of a base-paired stem and loop structure that is not closely bound to the remainder of the molecule. A survey of the results suggests that the V1 molecule may interact with the minor groove of the double helix with an affinity for stacked bases and that it may require two or three stacked bases for optimal binding and cleavage. PMID- 6291589 TI - In vitro transcription of the early region of Caulobacter phage phi Cd1 deoxyribonucleic acid by host RNA polymerase. AB - Transcription of the Caulobacter crescentus phage phi Cd1 genome requires both the host RNA polymerase and a phage-encoded, rifampicin-resistant RNA polymerase. Transcription of the early region of the phi Cd1 genome was examined in vitro with C. crescentus RNA polymerase. Four transcripts, A, B, C, and D, which ranged in size from 2.9 X 10(6) to 0.53 X 10(6) daltons, were synthesized in vitro by the holoenzyme. Transcript A appeared to be the major transcript since (a) it was the size of the entire 20% of the genome shown in vivo to code for the early phage mRNA, (b) it was one of the first transcripts synthesized at low enzyme-to DNA molar ratios, and (c) it was synthesized in approximately 3 times the molar equivalent observed for the other transcripts. The A transcript initiated primarily with GTP although a portion was also labeled with ATP. The B, C, and D transcripts were present in equivalent molar ratios, were all smaller than transcript A, and were found to yield RNase III digestion products that were subsets of each other as well as of transcript A. Each of these transcripts proved to be a de novo transcript since (a) each could be pulse labeled during the initial 20 s of the reaction and (b) each transcript contained a triphosphate at its 5' terminus. Evidence is presented that suggests that the B and C transcripts initiate at or near the major A promoter but terminate at different termination or pause sites within the early region of the phage genome. Transcript D appears to initiate at a minor promoter within the terminally redundant region of the genome preceding the A promoter. PMID- 6291590 TI - Characterization of the collagens synthesized by Chinese hamster ovary cells. Effect of colcemid and dibutyryladenosine cyclic monophosphate. AB - The collagens synthesized by Chinese hamster ovary cells have been isolated and characterized. Although these cells produce very small amounts of collagen, at least five distinct collagenous chains could be identified from radiolabeled media and cell extracts after limited pepsin digestion. Two chains were characterized as alpha 1(V) and alpha 2(V), based on electrophoretic mobility, resistance to vertebrate collagenase, chromatographic properties on carboxymethylcellulose, and cyanogen bromide peptide patterns. Two smaller collagenous proteins (Mr 34000 and 37000) were also isolated by carboxymethylcellulose chromatography and characterized by cyanogen bromide digestion patterns. These collagens showed similarities to type IV collagen fragments but may be unique to Chinese hamster ovary cells. A colcemid-resistant mutant of Chinese hamster ovary cells designated CMR795 [Ling, V., Aubin, J.E., Chase, A., & Sarangi, F. (1979) Cell (Cambridge, Mass.) 18, 423-430] was found to synthesize the same collagen chains but in different proportions. In the wild type cells colcemid (0.05-0.1 microgram/mL) reduced the amount of type V collagen in the culture media but had little effect on the other collagen type, whereas the type V collagen reduction was less pronounced in the CMR795 cells treated with the same concentrations of colcemid. Dibutyryladenosine cyclic monophosphate caused a fibroblast-like "reverse transformation" of the Chinese hamster ovary cells similar to that described previously [Hsie, A.W., & Puck, T. T. (1971) Proc. Natl. Acad. Sci. U.S.A. 68, 358-361]. However, collagen synthesis was increased only slightly. Furthermore, no apparent alteration in the types of collagens synthesized was detected. PMID- 6291591 TI - Reversible inhibition of the mitochondrial ubiquinol-cytochrome c oxidoreductase complex (complex III) by ethoxyformic anhydride. AB - The mitochondrial ubiquinol-cytochrome c oxidoreductase (complex III) is inhibited by ethoxyformic anhydride (EFA). The inhibition is readily reversed by hydroxylamine, suggesting the involvement of essential histidyl or possibly tyrosyl residues. The spectrum of ethoxyformylated complex III in the UV region showed a peak at 238 nm, indicative of N-(ethoxyformyl)histidine. Addition of hydroxylamine caused a large decrease of the 238-nm peak, which amounted to 16 mol of (ethoxyformyl)histidine/mol of cytochrome c1. Hydroxylamine addition to ethoxyformylated complex III also caused a small change at about 280 nm, which could be due to reversal of 1.6 O-ethoxyformylated tyrosyl residues/mol of cytochrome c1. Among many inhibitors of the cytochrome bc1 region of the respiratory chain, EFA is the only reagent known to cause reversible inhibition by covalent modification of amino acid residues. The inhibition site of EFA was determined to be between cytochromes b-562 and c1. However, unlike antimycin, which also inhibits in the same region, EFA did not promote the reduction of cytochrome b-566 in particles treated with substrates. In addition, it was found that EFA inhibits proton translocation in the cytochrome bc1 region and is a more effective electron transport inhibitor when added to reduced particles as compared to oxidized particles. These results together with the strong possibility that the EFA target is a histidyl or possibly a tyrosyl residue have been discussed in relation to the mechanism of proton translocation by complex III. PMID- 6291592 TI - Proton magnetic resonance studies of barley and wheat thionins: structural homology with crambin. AB - The thionins comprise a group of very basic proteins of Mr approximately 5000 found in the seeds of Gramineae. They each contain 45 amino acid residues arranged along a single polypeptide chain that is constrained by four disulfide bridges. Five thionins of known sequence, from barley and wheat, have been investigated and compared by 1H NMR spectroscopy at 600 MHz. From their spectral characteristics it is concluded that the five proteins have very similar, nonrandom conformations in 2H2O solution. Moreover, on the basis of selective nuclear Overhauser experiments at 300 MHz, features of their secondary and tertiary structures are shown to be similar to those of crambin, a related, hydrophobic protein extracted from seeds of the crucifer Crambe abyssinica. The strong compositional homology of the thionins facilitates the assignment of methyl and aromatic resonances, as only a few residues are replaced and these are at known sites. The substitution of leucine for an isoleucine does not affect significantly the local magnetic environment, suggesting that those isomeric side chains easily accommodate the same spatial constraints. A fast hydrogen-deuterium exchange is observed at pH 6.25, 25 degrees C. This indicates that, although of folded conformation, the thionins are structurally flexible polypeptides that efficiently expose all amides to the solvent. PMID- 6291593 TI - A combined proton and phosphorus-31 nuclear magnetic resonance investigation of the combining site of M603, a phosphocholine-binding myeloma protein. AB - Phosphorus-31 nuclear magnetic resonance (NMR) studies on the two phosphorus nuclei of the phosphonium analogue (Me3P+CH2CH2OPO3(2-)) of phosphocholine are used to monitor the charged subsites in the phosphocholine-binding immunoglobulin A mouse myeloma M603. Comparison of the 270-MHz 1H NMR difference spectrum on addition of either this analogue or phosphocholine to M603 and the almost identical changes in the pKa values of the phosphate groups on binding to M603 confirm that the analogue is a good model for phosphocholine. The pKa of the phosphate groups is decreased by 0.5 unit on binding to M603, which is consistent with the phosphate group being hydrogen bonding to Tyr-33H and Arg-95L, as suggested from the X-ray structure, and also implies that the binding energies for the mono- and dianion are similar. The P+Me3 moiety is used to probe the electrostatic interactions in the choline subsite. Titration of the chemical shift of the phosphonium phosphorus reflects a group on the protein that has a pKa value of less than or equal to 5, which from the refined X-ray structure (D.R. Davies, personal communication) of the site is assigned to Asp-97L. The choline subsite is monitored by using 1H NMR difference spectra, which indicates that the subsite is highly aromatic as expected from the crystal structure that places Trp-107H and Tyr-100L in this subsite. The ring current interactions from these rings can account for the 1H NMR chemical shift data on choline. PMID- 6291595 TI - Folding of the mitochondrial proton adenosinetriphosphatase proteolipid channel in phospholipid vesicles. AB - The mitochondrial H+-ATPase proteolipid from Neurospora crassa was incorporated into small unilamellar dimyristoylphosphatidylcholine vesicles and its conformation determined by circular dichroism spectroscopy (CD). While the largely alpha-helical conformation is relatively independent of the method of incorporation into vesicles, i.e., rehydration, detergent dialysis, or detergent dilution, the proteolipid conformation was significantly different in detergent micelles and in organic solvents. Only very slight changes in the CD spectrum were observed upon binding of the H+-ATPase inhibitor dicyclohexylcarbodiimide to the proteolipid in vesicles, thus suggesting that the inhibitor acts either by blocking the channel or by masking an essential charge group, rather by than causing an overall conformational change in the channel. Additionally, very similar CD spectra were obtained for vesicles with different lipid/protein mole ratios, indicating either that no substantial conformational differences exist between monomer and multimers or that monomers self-associate to form stable complexes during incorporation into vesicles. This study has provided a physical basis for model-building studies of the proteolipid channel structure. PMID- 6291594 TI - Activation of poly(adenosine diphosphate ribose) polymerase by SV 40 minichromosomes: effects of deoxyribonucleic acid damage and histone H1. AB - Poly(ADP-ribose) polymerase is a chromosomal enzyme that is completely dependent on added DNA for activity. The ability of DNA molecules to activate the polymerase appears to be enhanced by the presence of DNA damage. In the present study, we used SV 40 DNA and SV 40 minichromosomes to determine whether different types of DNA damage and different chromosomal components affect stimulation of polymerase activity. Treatment of SV 40 minichromosomes with agents or conditions that induced single-strand breaks increased their ability to stimulate poly(ADP ribose) synthesis. This stimulation was enhanced by addition of histone H1 at a ratio of 1 microgram of histone H1 to 1 microgram of DNA. Higher ratios of histone H1 to DNA suppressed the ability of SV 40 minichromosomes containing single-strand breaks to stimulate enzyme activity. Treatment of SV 40 minichromosomes or SV 40 DNA with HaeIII restriction endonuclease to produce double-strand breaks markedly stimulated poly(ADP-ribose) polymerase activity. The stimulation of poly(ADP-ribose) polymerase by double-strand breaks occurred in the absence of histone H1 and was further enhanced by adding histone H1 up to ratios of 2 to 1 relative to DNA. At higher ratios of histone H1 to DNA, the presence of the histone continued to enhance the poly(ADP-ribose) synthesis stimulated by double-strand breaks. PMID- 6291596 TI - Torsional motion and elasticity of the deoxyribonucleic acid double helix and its nucleosomal complexes. AB - Torsional thermal oscillations of the DNA double helix within the electron paramagnetic resonance (EPR) time scale (10(-10)-10(-3) s) as indicated by a rigid, intercalating probe are much smaller in the spacer segment between nucleosomes in chromatin than in long, free DNA molecules. Still smaller DNA oscillation is indicated in intact nuclei and yet smaller if the nuclei have been treated with glutaraldehyde. The values of EPR measurements are not affected by the loading density of probe. If the probe were capable of substantial oscillations or movement different from that of the helix, those oscillations would be expected to dominate the spectra when movement of the helix is restrained. We conclude that the correlation time for torsional movement of free DNA inferred from EPR spectra is characteristic of the double helix and that there is no significant independent motion of the probe. The correlation time for the DNA double helix in molecules longer than approximately 500 base pairs is close to 30 ns, corresponding to an elastic constant of 1.5 X 10(-19) ergs cm for deformation by twisting. The motions observed in chromatin are consistent with a model in which spheres of 50-60-A radius are connected by simple elastic rods with the length of spacer DNA and the same elastic constant. The spin-labeled ethidium probe has been characterized in detail by nuclear magnetic resonance, infrared, fluorescence, and visible light spectroscopy. The binding equilibria are consistent with the hypothesis that strongly immobilized probe molecules are preferentially bound to spacer DNA. PMID- 6291597 TI - Deoxyribonucleic acid-protein and deoxyribonucleic acid interstrand cross-links induced in isolated chromatin by hydrogen peroxide and ferrous ethylenediaminetetraacetate chelates. AB - DNA-protein and DNA interstrand cross-links were induced in isolated chromatin after treatment with H2O2 and ferrous ethylenediaminetetraacetate (EDTA). Retention of DNA on membrane filters after heating of chromatin in a dissociating solvent indicated the presence of a stable linkage between DNA and protein. Treatment of protein-free DNA with H2O2/Fe2+-EDTA did not result in enhanced filter retention. Incubation of cross-linked chromatin with proteinase K completely eliminated filter retention. Resistance to S1 nuclease after a denaturation-renaturation cycle was used to detect DNA interstrand cross-links. Heating the treated chromatin at 45 degrees C for 16 h and NaBH4 reduction enhanced the extent of interstrand cross-linking. The following data are consistent with, but do not totally prove, the hypothesis that cross-links are induced by hydroxyl radicals generated in Fenton-type reactions: (1) cross linking was inhibited by hydroxyl radical scavengers; (2) the degree of inhibition of DNA interstrand cross-links correlated very closely with the rate constants of the scavengers for reaction with hydroxyl radicals; (3) cross linking was eliminated or greatly reduced by catalase; (4) the extent of cross linking was directly related to the concentration of Fe2+-EDTA. Partial inhibition of cross-linking by superoxide dismutase indicates that superoxide driven Fenton chemistry is involved. The data indicate that DNA cross-linking may play a role in the manifestation of the biological activity of agents or systems that generate reactive hydroxyl radicals. PMID- 6291598 TI - A proton nuclear magnetic resonance investigation of histidyl residues in human normal adult hemoglobin. AB - High-resolution proton nuclear magnetic resonance (NMR) spectroscopy at 250 MHz has been used to titrate 22 individual surface histidyl residues (11 per alpha beta dimer) of human normal adult hemoglobin in both the deoxy and the carbon monoxy forms. The proton resonances of beta 2, beta 143, and beta 146 histidyl residues are assigned by a parallel 1H NMR titration of appropriate mutant and chemically modified hemoglobins. The pK values of the 22 histidyl residues investigated are found to range from 6.35 to 8.07 in the deoxy form and from 6.20 to 7.87 in the carbon monoxy form, in the presence of 0.1 M Bis-Tris or 0.1 M Tris buffer in D2O with chloride ion concentrations varying from 5 to 60 mM at 27 degrees C. Four histidyl residues in the deoxy form and one histidyl residue in the carbon monoxy form are found to have proton nuclear magnetic resonance titration curves that deviate greatly from that predicted by the simple proton dissociation equilibrium of a single ionizable group. The proton nuclear magnetic resonance data are used to ascertain the role of several surface histidyl residues in the Bohr effect of hemoglobin under the above-mentioned experimental conditions. Under these experimental conditions, we have found that (i) the beta 146 histidyl residues do not change their electrostatic environments significantly upon binding of ligand to deoxyhemoglobin and, thus, their contribution to the Bohr effect is negligible, (ii) the beta 2 histidyl residues have a negative contribution to the Bohr effect, and (iii) the total contribution of the 22 histidyl residues investigated here to the Bohr effect is, in magnitude, comparable to the Bohr effect observed experimentally. These results suggest that the molecular mechanism of the Bohr effect proposed by Perutz [Perutz, M.F. (1970) Nature (London) 228, 726-739] is not unique and that the detailed mechanism depends on experimental conditions, such as the solvent composition. PMID- 6291599 TI - A proton nuclear magnetic resonance investigation of histidyl residues in sickle hemoglobin. AB - Using high-resolution proton nuclear magnetic resonance spectroscopy at 250 MHz, we have determined the individual pK values of 22 surface histidyl residues (11 per alpha beta dimer) of sickle hemoglobin in both deoxy and carbon monoxy forms. Seven histidyl residues in the deoxy form and three in the carbon monoxy form are found to have pK values and chemical shifts different from the corresponding ones in human normal adult hemoglobin. Two of these histidyl residues are the beta 2 histidine and the beta 146 histidine, indicating that the conformations of the amino- and carboxyl-terminal regions of the beta chain in sickle hemoglobin are altered compared to those in human normal adult hemoglobin. The differences in the pK values of the additional surface histidyl residues between sickle and normal hemoglobins suggest that the effect of the amino acid substitution at the sixth position of the beta chain in sickle hemoglobin, namely, glutamic acid replaced by valine, is not restricted to the region around the mutation site but can extend to other regions in the protein molecule. In the deoxy form, the histidyl residues of sickle hemoglobin that have altered pK values and chemical shifts compared to the corresponding ones in human normal adult hemoglobin have been found to be sensitive to the early stages of the polymerization process [Russu, I.M., & Ho, C. (1980) Proc. Natl. Acad. Sci. U.S.A. 77, 6577-6581]. PMID- 6291600 TI - Effects of S-adenosyl-1,8-diamino-3-thiooctane on polyamine metabolism. AB - Exposure of mammalian cells (transformed mouse fibroblasts or rat hepatoma cells) to S-adenosyl-1,8-diamino-3-thiooctane produced profound changes in the intracellular polyamine content. Putrescine was increased and spermidine was decreased, consistent with the inhibition of spermidine synthase by this compound, which is a potent and specific "transition-state analogue inhibitor" of the isolated enzyme in vitro. The spermine content of the cells was increased by exposure to this drug presumably since spermine synthase was able to use a greater proportion of the available decarboxylated S-adenosylmethionine when spermidine synthase was inhibited. The decarboxylated S-adenosylmethionine content rose substantially because the activity of S-adenosylmethionine decarboxylase was increased in response to the decline in spermidine. These results indicate that S-adenosyl-1,8-diamino-3-thiooctane is taken up by mammalian cells and is an effective inhibitor of spermidine synthase in vivo and that S-adenosylmethionine decarboxylase is regulated by the content of spermidine, but not of spermine. The growth of SV-3T3 cells was substantially reduced in the presence of S-adenosyl-1,8-diamino-3-thiooctane at concentrations of 50 microM or greater. Such inhibition was reversed by the addition of spermidine but not by putrescine. When SV-3T3 cells were exposed to 5 mM alpha (difluoromethyl)ornithine and 50 microM S-adenosyl-1,8-diamino-3-thiooctane, the content of all polyamines was reduced. Putrescine and spermidine declined by more than 90% and spermine by 80%. Such cells grew very slowly unless spermidine was added. PMID- 6291601 TI - Effect of hydrogen ion concentration on rhodopsin-lipid interactions. PMID- 6291602 TI - Identification of two different Q-binding sites in QH2-cytochrome c oxidoreductase, using the Q analogue n-heptadecylmercapto-6-hydroxy-5,8 quinolinequinone. AB - The pK and mid-point redox potential of the Q-analogue 7-(n-heptadecyl)mercapto-6 hydroxy-5,8-quinolinequinone (HMHQQ) in aqueous medium are so low that under the experimental conditions used for studying the inhibition of electron transfer in submitochondrial particles only the oxidized, anionic form is present. The KD of the analogue, determined by comparing its inhibitory effect with that of n-heptyl 4-hydroxyquinoline N-oxide, is (0.003 + 0.24 x mg protein/ml) microM. The inhibition of succinate oxidation is pH dependent, due to a pH-dependent change in the overcapacity of the QH2-oxidizing system above the Q-reducing system. If the terminal part of the respiratory chain is reduced with ascorbate, the analogue inhibits the reduction of cytochrome b by substrate in the presence of antimycin with a similar KD value. In the absence of ascorbate the KD value is 100-times higher. The reduction of cytochrome b by substrate in particles treated with 2,3-dimercaptopropanol (BAL) + O2 is also sensitive to HMHQQ, with a KD value in between the two values given above. It is concluded that the QH2 oxidase system contains two different sites for interaction with ubiquinone. The site responsible for the inhibition of steady-state electron transfer is near the Fe-S cluster, as is shown by the sensitivity to the redox state of this cluster and by the effect of HMHQQ on the EPR signal of the reduced cluster. The second site, which is similar to the antimycin-binding site, is occupied only at higher concentrations of inhibitor. The affinity of HMHQQ for this site is not affected by the redox state of the Fe-S cluster. PMID- 6291603 TI - X-ray absorption spectroscopy. Application to biological molecules. PMID- 6291604 TI - Mitochondrial calcium transport. PMID- 6291605 TI - Alcohols inhibit adipocyte basal and insulin-stimulated glucose uptake and increase the membrane lipid fluidity. AB - Benzyl alcohol and ethanol, at aqueous concentrations that cause local anesthesia of rat sciatic nerve, affect structural and functional properties of rat adipocytes. The data strongly suggest that structurally-intact membrane lipids are required for the proper cellular uptake of glucose and for the physiologic response of adipocytes to insulin. The structure of adipocyte membrane lipids was examined with the spin label method. Isolated adipocyte 'ghost' membranes were labeled with the 5-nitroxide stearate spin probe I(12,3). Order parameters that are sensitive to the fluidity of the lipid environment of the incorporated probe were calculated from ESR spectra of labeled membranes. Benzyl alcohol and ethanol dramatically increased the fluidity of the adipocyte ghost membrane, as indicated by decreases in the polarity-corrected order parameter S. This concentration dependent fluidization commenced at approx. 10 mM benzyl alcohol and progressively increased at all higher concentrations tested (up to 107 mM). S decreased approx. 5.7% at 40 mM benzyl alcohol, a change in S comparable in magnitude to that induced by a 6 degrees C increase in the incubation temperature. Benzyl alcohol and ethanol inhibited basal glucose uptake in adipocytes and uptake maximally stimulated by insulin. Temperature-induced increases in membrane fluidity, detected with I(12,3), that closely paralleled the fluidity effects of alcohols were associated only with increases in basal and insulin-stimulated glucose uptake. The contention that the membrane lipid fluidity plays a role in insulin action needs further study. PMID- 6291606 TI - Validity of the Goldman-Hodgkin-Katz equation in paracellular ionic pathways of gallbladder epithelium. AB - The Goldman-Hodgkin-Katz equation has been extensively used to determine cationic/anionic permeability ratios in the paracellular pathways of the gallbladder epithelium. Nevertheless, new experimental evidence suggests that none of the theoretical assumptions of the equation hold for these pathways. In order to assess the experimental validity of the Goldman equation the permeability ratios were calculated from zero-current diffusion potentials by means of the Goldman equation and compared with the cationic/anionic permeability ratios measured by simultaneous determinations of cation and anion tracer fluxes in the same membranes. The results indicate that the Goldman equation is empirically valid for the tested salts (KCl and RbCl) within the experimental range of concentrations (25 to 200 mM) at an electrochemical-potential difference of zero. PMID- 6291607 TI - An improved procedure for the preparation and measurement of (Na+ + K+)-ATPase in human erythrocytes. PMID- 6291608 TI - Electron microscopic study on the interaction of Sendai virus with liposomes containing glycophorin. AB - The interaction of liposomes containing glycophorin, a major sialoglycoprotein of human erythrocytes, with Sendai virus was studied by freeze-fractures and negative staining electron-microscopy. Viral envelopes were absorbed on liposomal membranes at 0 degrees C. When the temperature was shifted up to 37 degrees C, the viral envelopes fused with the liposomal membranes (envelope fusion). Particles representing viral membrane components formed clusters on liposomal membranes after incubation for more than 1 h at 37 degrees C. PMID- 6291609 TI - An electrophoretic analysis of proteolipids from different rat brain subcellular fractions. AB - Proteolipid proteins were extracted from adult rat brain subcellular fractions and purified by chromatography on Sephadex LH-60. Polyacrylamide gel electrophoresis of the delipidized proteins, in the presence or absence of 8 M urea, was carried out with all fractions. The distribution of the various types of proteolipid proteins was studied and their molecular weight calculated by the Ferguson relationship. Several bands of proteolipid proteins were found in the five membrane fractions analyzed. Some of them, such as the 17.5 K and 37 K components were very prominent in mitochondria and synaptosomes. The 30 K component was found in myelin-derived membranes and in microsomes, while the 20 K and 25 K proteolipid proteins were present in all subcellular fractions. The 30 K component (proteolipid protein (PLP)), typical of the purified myelin membranes, showed a similar distribution to that of 2',3'-cyclic-nucleotide 3' phosphohydrolase (EC 3.1.4.37) activity, while the other major proteolipid protein present in all subcellular fractions (25 K) did not show such parallelism, indicating that it might not be an exclusive component of myelin. The electrophoretic pattern of microsomal proteolipid proteins did not show the high molecular weight components (aggregates of PLP) which are found in myelin. Furthermore, the 30 K component showed a smaller Y0 value than that of the 30 K found in myelin. Thus the presence of 30 K proteolipid protein in microsomes should not be considered as being due to myelin contamination. PMID- 6291610 TI - K+ - and Na+ -gradient-dependent transport of L-phenylalanine by mouse intestinal brush border membrane vesicles. PMID- 6291611 TI - The affinity of the Ca2+ pump of human erythrocytes for external Na+ or K+. PMID- 6291612 TI - Studies of ions and water in human lymphocytes. PMID- 6291613 TI - An analysis of the repair processes in ultraviolet-irradiated Micrococcus luteus using purified ultraviolet-endonuclease. AB - The measurement of the frequency of endonucleolytic incisions in ultraviolet irradiated DNA serves as the test for the presence of pyrimidine dimers. In accordance with this approach, the lysates of three Micrococcus luteus strains containing radioactively labeled chromosomes were treated with purified M. luteus ultraviolet-endonuclease to trace segregation of dimers amongst parental and newly synthesized DNA and their removal during postreplication and excision DNA repair. A considerable proportion of the dimers in all strains tested proved to be insensitive to the action of exogenous incising enzyme. The use of chloramphenicol as an inhibitor of postirradiation protein synthesis in combination which ultraviolet-endonuclease treatment of DNA allowed to reveal at least two alternative pathways of postreplication repair: constitutively active recombinational pathway and inducible nonrecombinational one. PMID- 6291614 TI - The complete amino acid sequence of Nitrobacter agilis cytochrome c-550. AB - The amino acid sequence of cytochrome c-550 from the chemoautotroph, Nitrobacter agilis, was completed by using solid-phase sequencing and conventional procedures. The cytochrome was composed of 109 amino acid residues and its molecular weight was calculated to be 12375 including haem c. The cytochrome was homologous to eukaryotic cytochromes c and some photosynthetic bacterial cytochromes c2. In particular, its primary structure was very similar to that of Rhodopseudomonas viridis cytochrome c2. Some of its properties were compared with those of other cytochromes c on the basis of the primary structure. PMID- 6291615 TI - Effects of temperature, pH and detergents on the molecular conformation of the enterotoxin of Clostridium perfringens. AB - The effects of temperature, pH and sodium dodecyl sulfate on the conformation of the enterotoxin from Clostridium perfringens type A were followed by circular dichroism in both the peptide and aromatic regions. At near-physiological conditions (35 degrees C, pH 6.7) the enterotoxin exhibited a conformation consisting of approximately 60% pleated sheet, 40% non-periodic, and essentially no helix. The peptide region was relatively stable at temperatures up to 55 degrees C and at pH values ranging from 4-10. The aromatic region demonstrated profound, time-dependent changes at 55 degrees C. At temperatures greater than 55 degrees C, extremes of pH, and in the presence of SDS, the spectra in both regions showed major structural reorganization; in most cases a gain in helical content at the expense of sheet structure was observed. The conformational properties of the protein are very similar to those observed for the lectins, a group of carbohydrate-binding proteins. PMID- 6291616 TI - Na+, K+-dependent adenosine triphosphate phosphohydrolase. Two types of kinetics. AB - Kinetic analysis of hydrolytic activity of (Na+, K+)-ATPase purified from duck salt glands shows that several substrates are hydrolysed in different manners. UTP, GTP and ITP are hydrolysed in accordance with usual Michaelis kinetics with the single Km value and with no cooperatively (Hill coefficient, nH = 1), while CTP is hydrolysed, like ATP, in accordance with non-Michaelis kinetics with two Km values. Hydrolysis of the last two substrates in the range of the second Km is characterised by positive cooperativity with nH greater than 1. PMID- 6291617 TI - Differential acid stabilities of citraconylated amino groups of glucagon. Preparation of N alpha-citraconyl glucagon and evaluation of its biological properties. AB - Acylation of the alpha- and epsilon-amino groups of histidine-1 and lysine-12 in glucagon with citraconic anhydride resulted in the formation of amide bonds which displayed different stabilities to hydrolysis under mild acid conditions. Treatment of N alpha,epsilon-dicitraconyl glucagon at pH 4.0 and room temperature regenerated the free epsilon-amino group within 16 h, while the citraconyl-alpha amino group was stable. N alpha-Citraconyl glucagon was purified by anion exchange chromatography and was a weak partial agonist in stimulating adenylate cyclase in rat liver plasma membranes. The derivative exhibited 1% of the biological potency and 35-40% of the maximal stimulation of glucagon. Binding affinity to plasma membranes was also reduced, but not to as great an extent as adenylate cyclase activity. Removal of the alpha-citraconyl group by treatment with 10 mM HCl at 40 degrees C restored full potency and stimulation to glucagon. These results suggest that the N-terminal histidine of glucagon is involved in both binding to plasma membranes and transduction of the signal to adenylate cyclase. PMID- 6291618 TI - Induction of choline kinase by polycyclic aromatic hydrocarbons in rat liver. II. Its relation to net phosphatidylcholine biosynthesis. AB - The effect of a single dose (50 mg/kg body weight) of 3-methylcholanthrene on de novo phosphatidylcholine biosynthetic activities in rat liver was studied both in a cell-free system and with slice experiments. 3-Methylcholanthrene caused a significant depression of either [methyl-14C]choline or [2-(3)H]glycerol incorporation into phosphatidylcholine when the precursor was incubated with liver slices. At the same time, there occurred a significant accumulation of radioactivity in either cholinephosphate or diacylglycerol molecule from [14C]choline or [3H]glycerol, respectively, suggesting that 3-methylcholanthrene could cause an inhibitory effect on hepatic phosphatidylcholine synthesis at the cholinephosphotransferase or/and cholinephosphate cytidylyltransferase step. Subsequent studies, where the activities of the three enzymes involved in de novo phosphatidylcholine synthesis were compared between control and 3 methylcholanthrene-pretreated rat liver subcellular fractions, demonstrated that the cholinephosphotransferase step could be the site of inhibition by 3 methylcholanthrene. On the other hand, 3-methylcholanthrene caused a significant induction of choline kinase activity in a time-dependent manner and, at the same time, the cholinephosphate pool size in liver cytosol was enlarged 2-3-fold when compared to the respective control. The overall results suggested strongly that 3 methylcholanthrene causes the counteractive effects on the de novo phosphatidylcholine biosynthesis, induction of choline kinase activity and inhibition of cholinephosphotransferase activity, both of which could participate in a concomitant increase in cholinephosphate pool size in rat liver. PMID- 6291619 TI - Induction of choline kinase by polycyclic aromatic hydrocarbons in rat liver. I. A comparison of choline kinases from normal and 3-methylcholanthrene-induced rat liver cytosol. AB - Choline kinase in rat liver has been shown to be induced up to 2-fold by the administration of polycyclic aromatic hydrocarbon carcinogens such as 3 methylcholanthrene and 3,4-benzo[a]pyrene (Ishidate, K., Tsuruoka, M. and Nakazawa, Y., (1980) Biochem. Biophys. Res. Commun. 96, 946-952). In order to characterize the nature of choline kinase induction by these carcinogens, the 3 methylcholanthrene-induced form as well as the normal form of choline kinase were partially purified from rat liver cytosol through acid treatment, (NH4)2SO4 precipitation and DEAE-cellulose column chromatography with linear KCl-gradient elution, and the catalytic properties were compared between the two preparations. Both enzyme activities were purified about 17-fold with a yield of 50% through the purification steps and there appeared no detectable difference in the elution pattern from either DEAE-cellulose column or Sephadex G-200 gel filtration. On the other hand, some differences were observed in catalytic properties between the two enzyme preparations; (1) the induced form showed a higher apparent Km value for choline (0.19 mM) when compared to the normal form (0.11 mM) and (2) the addition of polyamines caused a considerable increase in the maximum reaction velocity for the normal form whereas no remarkable change for the induced form, when the activities were plotted as a function of choline concentration. The overall results suggest that the 3-methylcholanthrene-induced form of choline kinase in rat liver could be different from the normal form, or that there exist several isoenzymes of choline kinase in rat liver, and one or some of them are inducible by the administration of polycyclic aromatic hydrocarbons. PMID- 6291620 TI - Human milk peroxidase is derived from milk leukocytes. AB - Peroxidase enzymes present in human colostrum, saliva, polymorphonuclear leukocytes, and bovine milk were compared with respect to their molecular exclusion chromatographic behavior and immunological cross-reactivity. Human milk peroxidase gave an elution profile similar to myeloperoxidase derived from blood polymorphonuclear leukocytes. Human salivary peroxidase reacted with an antibody directed against bovine lactoperoxidase, but with the same antibody preparation no reaction was detected either with human milk peroxidase or leukocyte myeloperoxidase. We conclude that the peroxidase enzyme in human milk is different from the human salivary and the bovine enzymes and is probably derived from milk leukocytes. PMID- 6291621 TI - Dissociation of receptor-bound human chorionic gonadotropin from rat testicular membranes in vitro as a high molecular weight complex and its inhibition by heavy metals and alkylating agents. PMID- 6291622 TI - Prostacyclin prolongs viability of washed human platelets. AB - The functional viability of stored human platelets, washed in the presence and absence of prostacyclin, was examined over a 96 h period. Platelet counts, aggregation responses and cyclic AMP levels were monitored as well as the spontaneous generation of thromboxane B2 and the liberation of labelled oleate from cellular phosphatides. In suspensions prepared without prostacyclin in the medium, platelet counts declined rapidly as did the sensitivity to aggregating agents. In addition, substantial amounts of thromboxane B2 were generated during preparation and storage and oleate liberation occurred at a rapid rate. In contrast, in prostacyclin-washed platelets, aggregation was maintained throughout the study period and there was little generation of thromboxane B2. Moreover, only a gradual decrease in platelet count and a slow increase in the rate of oleate liberation was observed when compared with controls. However, cyclic AMP levels rapidly declined when platelets were resuspended in prostacyclin-free medium. PMID- 6291623 TI - Properties of enzyme activities involved in protein phosphorylation dephosphorylation of thyroid plasma membranes. AB - Bovine thyroid tissue exhibited cAMP-dependent and Ca2+-dependent protein kinase activities as well as a basal (cAMP- and Ca2+-independent) one, and phosphoprotein phosphatase activity. Although the former two protein kinase activities were not clearly demonstrated using endogenous protein as substrate, they were clearly shown in soluble, particulate and plasma membrane fractions using exogenous histones as substrate. The highest specific activities were in the plasma membrane. The apparent Km values of cAMP and Ca2+ for the membrane bound protein kinase were 5 . 10(-8) M and 8.3 . 10(-4) M in the presence of 1 Mm EGTA), respectively. The apparent Km values of Mg2+ were 7.10-4M (without (in the cAMP and Ca2+), 5 . 10(-4) M (with cAMP) and 1.3 . 10(-3) M (with Ca2+), and those of ATP were 3.5 . 10(-5)M (with or without cAMP) and 8.5 . 10(-5) M (with Ca2+). The Ca2+-dependent protein kinase could be dissociated from the membrane by EGTA-washing. The enzyme activity so released was further activated by added phospholipid (phosphatidylserine/1,3-diolein), but not by calmodulin. Phosphoprotein phosphatase activity was also clearly demonstrated in all of the fractions using 32P-labeled mixed histones as substrate. The activity was not modified by either cAMP or Ca2+, but was stimulated by a rather broad range (5-25 mM) of Mg2+ and Mn2+. NaCl and substrate concentrations also influenced the activity. Pyrophosphate, ATP, inorganic phosphate and NaF inhibited the activity in a dose-dependent manner. Trifluoperazine, chlorpromazine, dibucaine and Triton X-100 (above 0.05%, w/v) specifically inhibited the Ca2+-dependent protein kinase in plasma membranes. Repetitive phosphorylation of intrinsic and extrinsic proteins by the membrane-bound enzyme activities clearly showed an important co ordination of them at the step of protein phosphorylation. These findings suggest that these enzyme activities in plasma membranes may contribute to regulation of thyroid function in response to external stimuli. PMID- 6291624 TI - Roles of GTP and GDP in the regulation of the thyroid adenylate cyclase system. AB - Effects of guanine nucleotides on the adenylate cyclase activity of thyroid plasma membranes were investigated by monitoring metabolism of the radiolabeled nucleotides by thin-layer chromatography (TLC). When ATP was used as substrate with a nucleotide-regenerating system, TSH stimulated the adenylate cyclase activity in the absence of exogenous guanine nucleotide. Addition of GTP or GDP equally enhanced the TSH stimulation. Effects of GTP and GDP were indistinguishable in regard to their inhibitory effects on NaF-stimulated activities. The results from TLC suggested that GDP could be converted to GTP by a nucleotide-regenerating system. Even in the absence of a nucleotide regeneration system, addition of GDP to the adenylate cyclase assay mixture resulted in the parallel decrease in ATP levels and formation of GTP indicating that thyroid plasma membrane preparations possessed a transphosphorylating activity. When an ATP analog, App[NH]p, was used as substrate without a nucleotide-regenerating system, no conversion of GDP to GTP was observed. Under such conditions, TSH did not stimulate the adenylate cyclase activity unless exogenous GTP or Gpp[NH]p was added. GDP no longer supported TSH stimulation and caused a slight decrease in the activity. GDP was less inhibitory than Gpp(NH)p to the NaF-stimulated adenylate cyclase activity. These results suggest: (1) TSH stimulation of thyroid adenylate cyclase is absolutely dependent on the regulatory nucleotides. (2) In contrast to GTP, GDP cannot support the coupling of the receptor-TSH complex to the catalytic component of adenylate cyclase. (3) The nucleotide regulatory site is more inhibitory to the stimulation of the enzyme by NaF when occupied by Gpp[NH]p than GDP. PMID- 6291625 TI - Effect of oxygen radicals and hyperoxia on rat heart ornithine decarboxylase activity. AB - Rat heart ornithine decarboxylate activity from isoproterenol-treated rats was inactivated in vitro by reactive species of oxygen generated by the reaction xanthine/xanthine oxidase. Reduced glutathione, dithiothreitol and superoxide dismutase has a protective effect in homogenates and in partially purified ornithine decarboxylase exposed to the xanthine/xanthine oxidase reaction, while diethyldithiocarbamate, which is an inhibitor of superoxide dismutase, potentiated the damage induced by O2- on enzyme activity. Dithiothreitol at concentrations above 1.25 mM had an inhibitory effect upon supernatant ornithine decarboxylase activity, while at 2.5 mM it was most effective in the recovery of ornithine decarboxylase activity, after the purification of the enzyme by the ammonium sulphate precipitation procedure. The ornithine decarboxylase inactivated by the xanthine/xanthine oxidase reaction showed a higher value of Km and a reduction of Vmax with respect to control activity. The exposure of rats to 100% oxygen for 3 h reduced significantly the isoproterenol-induced heart ornithine decarboxylase activity. The injection with diethyldithiocarbamate 1 h before hyperoxic exposure further reduced heart ornithine decarboxylase activity. PMID- 6291626 TI - Effect of hormones and 3', 5'-cyclic AMP on very low density lipoprotein receptors. AB - Human 125I-labelled VLDL interacts with rat adipocytes in vitro, with properties typical of a ligand-receptor interaction. This VLDL-receptor interaction is modulated by hormones which are known to change cyclic AMP levels. Norepinephrine and isoproterenol, both of which elevate cycle AMP, increase the binding of VLDL to adipocytes. Dibutyryl-cyclic AMP, a derivative of cyclic AMP, also increases the VLDL binding to adipocytes. Insulin reverses the catecholamine-induced increase in VLDL binding. This parallels insulin's effect on the catecholamine induced changes in cyclic AMP. Direct addition of cyclic AMP itself increases VLDL binding to adipocyte membranes, a system in which no lipolysis of new protein synthesis occurs. Based on the competition between unlabelled VLDL and 125I-labelled VLDL, we conclude that catecholamines act on adipocytes, and cyclic AMP on membrane fractions, by increasing their capacity rather than their affinity to bind VLDL. PMID- 6291627 TI - Vitamin D receptors in isolated rat parotid gland acinar cells. AB - Rat parotid gland was examined for the presence of 1 alpha, 25 dihydroxycholecalciferol receptors using sucrose density gradient ultracentrifugation techniques. [3H] DHCC bound specifically and with high affinity to a 3.2 S protein present in nuclear and cytosolic fractions of isolated parotid acinar cells. Values for the equilibrium dissociation constant and for the receptor concentration were determined to be approx. 0.1 nM, and 12 fmol/mg protein, respectively. In competitive inhibition experiments, the 3.2 S protein displayed 100-fold lower affinity for 25-hydroxycholecalciferol than for DHCC, and did not bind estradiol or methylprednisolone. These results suggest that rat parotid gland acinar cells contain classical DHCC receptors. A similar approach failed to provide evidence of DHCC receptors in isolated pancreas acinar cells, lacrimal gland or submandibular gland. It has been previously reported that vitamin D is essential for normal exocrine secretion from the rat parotid gland (Tenenhouse, A. and Afari, G. (1978) Biochim. Biophys. Acta 538, 631-634). The present findings suggest that this effect is the result of a direct action of DHCC on the parotid gland acinar cell. The absence of DHCC receptors in other exocrine cells suggests that tissue sensitivity to DHCC is not a general property of exocrine systems. PMID- 6291628 TI - Role of the adenylate system and glycolytic flux in the control of protein synthesis in isolated rat lung cells. AB - (1) Glucose stimulates the incorporation of amino acids into protein in lung cells isolated by digestion of the lung stroma with collagenase. This effect reflects mainly an increase in protein synthesis since no effect of glucose had been found to the uptake of amino acid precursors and, although glucose decreases the rate of intracellular proteolysis by 15%, this effect cannot account for the increased incorporation of radioactivity into proteins. Furthermore, glucose did not induce any significant change in the intracellular content of valine. (2) For glucose to act on protein synthesis, it must be glycolyzed since its stereoisomer, L-glucose, which is not metabolized by lung cells, has no effect. (3) The mechanism of glucose action does not seem to be related simply to variations of cellular ATP content or energy charge. The following arguments seem to support this conclusion: (i) glucose does not bring about significant variations in the concentration of reactants of the adenylate system; (ii) the increase in protein synthesis induced by glucose in energy-depleted cells correlates with a rise in ATP content and energy charge; however, adenosine, which increases ATP levels in a form quantitatively similar to glucose, is unable to affect protein synthesis: (iii) glucose also accelerates the incorporation of amino acids into proteins in adenosine-treated lung cells in which the ATP concentration was almost double that of the control and the energy charge was considerably elevated, ruling out the possibility that a rise in the steady-state concentration of ATP and/or energy charge alone could be responsible for the acceleration of protein synthesis. (4) It can be concluded that the effect of glucose in increasing protein synthesis in lung cells is dependent on some signal arising from its breakdown and not to variations in the concentration of reactants or energy charge of the adenylate system. PMID- 6291630 TI - [Thermodynamic characteristics of the interaction of high-potential cytochrome with dimer of bacteriochlorophyll in the reaction centers of Ectothiorhodospira shaposhnikovii chromatophores]. AB - Kinetics of dark reduction of cytochrome CH (E7=290 mV) after its photoinduced oxidation chromatophores of E. shaposhnikovii at 110-210 K is analysed. A ratio is derived which describes dark reduction of cytochrome. This ratio makes it possible to determine the value of free energy of the electron transfer between cytochrome and bacteriochlorophyll dimer of the reaction centre (Bchl)2. The values of enthalpy and entropy of the transition under study at the temperatures below 210 K are equal to 1,67 kJ M-1 and 26,8 J M-1 K-1 correspondingly. A change of free energy obtained by linear extrapolation to the room temperature equals 9,71 kJ M-1. This value well agrees with the value 9,63 kJ M-1 obtained from the results of direct potentiometric titration of photoinduced redox transformations CH and (Bchl)2. PMID- 6291629 TI - Degradation of glucagon receptors by dispase treatment of isolated intact rat hepatocytes. AB - Collagenase-isolated rat hepatocytes were treated with dispase II, a neutral proteolytic enzyme which is often used for the disintegration of neonatal cells. The treatment of hepatocytes with dispase II caused a significant reduction of glucagon binding to the intact cells. The deleterious effect of this enzyme on the specific glucagon binding site is accompanied by a reduction of the maximum intracellular cyclic AMP production. PMID- 6291631 TI - [Incorporation of sarcoplasmic reticulum membrane in planar bilayer]. PMID- 6291632 TI - [Change of mechanical parameters as a possible means for information processing by the neuron]. AB - The depolarization of neuronal membrane during cAMP injection by means of both iontophoresis and pressure was shown. Neuron blowing by solutions without cAMP gave the similar effect. The effect of cAMP can be differentiated in the presence of phosphodiesterase inhibitors. The experiments support the hypothesis that cyclic nucleotides and mechanical excitation interact with intraneuronal mechanical analog device (MAD) which consists of microtubules and microfilaments. It is supposed that MAD controlled by cyclic nucleotides and Ca2+ is the intraneuronal information processing system. PMID- 6291633 TI - [Scorpion toxins: an example of the use of venomous secretions]. PMID- 6291634 TI - Properties and regulation of the UVRABC endonuclease. AB - This report summarizes the cloning of the uvrA, uvrB and uvrC genes of E. coli, the identification and isolation of the gene products, the regulation of the genes, and reconstitution of active UVRABC endonuclease from the individually isolated components. PMID- 6291635 TI - Mutagenesis and other responses induced by DNA damage in Escherichia coli. PMID- 6291637 TI - Ultraviolet-irradiated simian virus 40 activates a mutator function in rat cells under conditions preventing viral DNA replication. AB - The UV-irradiated temperature-sensitive early SV40 mutant tsA209 is able to activate at the nonpermissive temperature the expression of mutator and recovery functions in rat cells. Unirradiated SV40 activates these functions only to a low extent. The expression of these mutator and recovery functions in SV40-infected cells was detected using the single-stranded DNA parvovirus H-1 as a probe. Because early SV40 mutants are defective in the initiation of viral DNA synthesis at the nonpermissive temperature, these results suggest that replication of UV damaged DNA is not a prerequisite for the activation of mutator and recovery functions in mammalian cells. The expression of the mutator function is dose dependent, i.e., the absolute number of UV-irradiated SV40 virions introduced per cell determines its level. Implications for the interpretation of mutation induction curves in the progeny of UV-irradiated SV40 in permissive host cells are discussed. PMID- 6291636 TI - recA operator mutations and their usefulness. AB - Mutations at three positions in the operator for recA have been detected, cloned and sequenced. Derepressed amounts of recA protein vary over a forty-fold range and correlate well with reduced affinities for lexA repressor of the mutant operators. One mutant confirms the region of major groove interaction between repressor and operator. Another has been used to demonstrate that RecF pathway genes other than recA are under lexA control. PMID- 6291638 TI - DNA-damage inducible genes on the I group plasmid TP 110. PMID- 6291639 TI - Enhanced reactivation and mutagenesis after transfection of carcinogen-treated monkey kidney cells with UV-irradiated simian virus 40 (SV40) DNA. AB - Monkey kidney cells, either untreated or pretreated with UV-light at 254 nm or mitomycin C, were transfected 24 hours later with the intact or UV-irradiated DNA from the thermosensitive tsB201 simian virus 40 mutant unable to grow at 41 degrees C. The survival of the viral progeny obtained from the UV-irradiated DNA is increased in pretreated cells compared to the survival of the viral progeny obtained in untreated cells. Irradiation of the viral DNA enhances the reversion frequency of the viral progeny towards a wild type phenotype able to grow at 41 degrees C. Pretreatment of the cells with UV or mitomycin C does not increase the reversion frequency. PMID- 6291640 TI - Mutational specificity of the umuC mediated mutagenesis in Eschericha coli. PMID- 6291641 TI - Error-prone replication of ultraviolet-irradiated simian virus 40 in carcinogen treated monkey kidney cells. AB - To analyze the molecular mechanism of mutagenesis in carcinogen-treated mammalian cells, we developed a model system composed of various simian virus 40 (SV40) mutants as a biological probe to detect inducible DNA repair and mutagenesis in carcinogen-treated monkey kidney cells (CV1-P). Results have shown that treatment of cells with UV-light, acetoxy-acetyl-aminofluorene, or mitomycin C, increases the mutagenesis of UV-irradiated SV40 ts mutant measured as a reversion frequency from a thermosensitive phenotype toward a thermoresistant phenotype. This increased mutagenesis is not observed in the case of undamaged virus indicating that we are looking at targeted mutagenesis. The molecular analysis of several revertant genomes indicates that some DNA rearrangements may occur in the revertant genomes but in some cases the reversion site is a single basepair substitution located at positions different from the original thermosensitive mutation, which is still present. The general interpretation of our results leads to the conclusion that carcinogen treatment of monkey cells activates some kind of error-prone replication mode able to better replicate UV-damaged templates but leading to a higher level of mutagenesis. This activity may represent a SOS-like function in mammalian cells. PMID- 6291642 TI - Two separable protein species which both restore uvrABC endonuclease activity in extracts from uvrC mutated cells. AB - Two different protein species which both complement the detective repair endonuclease (uvrABC endonuclease) in uvrC mutated cells have been detected. These proteins have quite different chromatographic properties and were easily separated by ion exchange chromatography. One has affinity for DEAE cellulose and co-cromatographs with the uvrB protein. The other has strong affinity for phosphocellulose and appears to be the uvrC protein itself. The uvrB associated uvrC+ activity is absent from both uvrC and uvrB mutated cells, indicating that this species result from an interaction between uvrB+ and uvrC+ functions at the protein level. PMID- 6291643 TI - [Inhibition of H2O2 and O2-. generation in the respiratory chain, treated with 2,3-dimercaptopropanol]. AB - Inhibition of electron transport in bovine heart submitochondrial particles resulting from treatment with British antilewisite is accompanied by suppression of superoxide radical and H2O2 generation at the b-c1 site of the respiratory chain. In conjunction with the previously obtained data, the results suggest that the effect of British antilewisite is directed towards the step of ubihydroqinone oxidation to ubisemiquinone by the Rieske FeS-protein. PMID- 6291644 TI - [Action of the venom of the spider Lityphantes paykullianus on synaptic processes]. PMID- 6291645 TI - Effect of carbamazepine on cyclic nucleotides in CSF of patients with affective illness. PMID- 6291646 TI - Ion transport and adrenergic function in major affective disorder. AB - Altered functional levels of norepinephrine (NE) have been implicated in the etiology of affective disorders. Abnormalities of membrane ion transport have been postulated to underlie such neurotransmitter imbalances. To evaluate the role of these systems in the pathophysiology of specific affective syndromes, we have examined the NE metabolite MHPG and ion transport function in a cross section of affective patients in different mood states. Analysis of those data reveals that plasma MHPG levels are dependent on mood state, with bipolar patients in the manic phase having significant elevations of this metabolite. In contrast, the red cell: plasma lithium ratio (LR) was independent of mood state and significantly elevated in a group of bipolar patients. PMID- 6291647 TI - Red cell ouabain-sensitive Na+-K+-adenosine triphosphatase: a state marker in affective disorder inversely related to plasma cortisol. AB - Ouabain-sensitive Na+-K+-stimulated ATPase was measured in red cell membranes using a spectrophotometric assay. The mean enzyme level in patients in the depressed state (1.2 nM/mg protein per min +/- 0.18 SEM) was lower lower than that in well-state patients (2.0 +/- 0.26) and hypomanic patients (2.4 +/- 0.31). Lithium treatment itself did not alter ATPase levels. Levels in patients in the well state were not significantly different from controls and thus ATPase does not constitute a "trait" marker for affective illness. Plasma cortisol level was higher in well-state patients (15.9 micrograms/dl +/- 1.46) than in controls (11.5 +/- 0.75). There were no significant differences in cortisol in these single morning samples during different mood states. Cortisol level correlated negatively with ATPase level in the total group of patients (r = 0.42, p less than 0.005), especially in those who were euthymic. These data indicate a relationship between cortisol and ATPase levels in affectively ill patients. Ouabain-sensitive NaK ATPase may be useful as an indicator of state in affective illness; plasma cortisol may be continuously elevated in some individuals with affective disorder. PMID- 6291649 TI - Monolayer growth and differentiated function of porcine and rat granulosa cells following cryopreservation. PMID- 6291648 TI - Stress-induced testicular hyposensitivity to gonadotropin in rats. Role of the pituitary gland. AB - The time course of stress-induced testicular hyposensitivity to gonadotropins was studied in hypophysectomized or naloxone-treated rats exposed to various periods of immobilization. Blood was collected from a chronically indwelling intra-atrial catheter every hour for luteinizing hormone (LH) and testosterone (T) measurement. Eight hours of immobilization completely suppressed T secretion without significant effect on LH. Human chorionic gonadotropin (hCG, 5 IU/rat, i.m.) induced a marked increase in plasma T levels in normal control groups 3 h post-injection while in immobilized rats the response was completely abolished, even after only 30 min of stress. In hypophysectomized rats, as expected, plasma T levels were undetectable, but, contrary to results obtained in normal animals, hCG induced a similar increase of plasma T levels both in control and stressed rats. Immobilization stress failed to inhibit plasma T values in hypophysectomized rats pretreated for 4 days with human menopausal gonadotropin (hMG) + hCG, while it did so in similarly treated normal animals. Naloxone induced a rise of plasma LH and T levels in control rats, but did not antagonize the stress-induced fall of plasma T concentration. In all groups, steroid testicular content mimicked variations of plasma T values. In particular, in stressed animals the lack of accumulation of testicular 17-hydroxyprogesterone probably reflected a normal activity of 17-20 lyase. These results indicate that stress induces very rapidly a state of Leydig cell hyposensitivity to gonadotropins and a blockade of T biosynthesis. The causal relationship between the two effects is presently not clear but these events seem to be due to stress induced release of an inhibitory factor of pituitary origin other that endorphin. PMID- 6291650 TI - Localization of relaxin binding sites in the rat uterus and cervix by autoradiography. AB - 125I-labeled porcine relaxin was injected into 27-day-old rats treated with pregnant mare's serum gonadotropin (PMSG) and known target tissues for relaxin, the myometrium, endometrium and cervix, and putative control tissues, heart, thigh muscle and duodenum, examined for binding by autoradiography. Specific binding in the target tissues was demonstrated by simultaneous injection of excess unlabeled relaxin. Radioactivity was located and quantified by grain counts predominantly over the inner, circular muscle layer of the myometrium and the cervix and to a lesser extent over the outer longitudinal muscle layer of the myometrium and the endometrium. The route of injection, the circulation time, or counting grains in transverse or longitudinal sections of myometrium made little difference in these results. Ovariectomy decreased, but not significantly, the grain count in all of the target tissues studied and estrogen treatment of ovariectomized animals restored the numbers of grains to approximately that of intact PMSG-treated rats. The degree of binding of the cervix was approximately that of the circular myometrial muscle. This work confirms the presence of specific receptors for relaxin in the rat uterus and cervix of primed rats and it also suggests that the inhibitory action of relaxin upon the myometrium is primarily on the inner circular muscle layer. PMID- 6291651 TI - Characterization of two steroidogenic cell types in the ovine corpus luteum. PMID- 6291652 TI - Yield, characteristics, motility and cAMP content of sperm isolated from seven regions of ram epididymis. PMID- 6291653 TI - Beta-endorphin is present in the male reproductive tract of five species. AB - Previous studies from this laboratory have demonstrated immunostainable beta endorphin-like material (beta-EP) in Leydig cells and epithelia of the epididymis, seminal vesicle and vas deferens of the rat. These observations would be strengthened if it could be demonstrated that they were not a peculiarity of the rat. Accordingly, we now present immunocytochemical evidence for the presence of beta-EP in the Leydig cells of mouse, hamster, guinea pig and rabbit. No immunoreactive material was identified in Sertoli, myoid, endothelial or germ cells of any of the species examined. Immunostainable beta-EP was also demonstrated in the epididymides of mouse, guinea pig, rabbit, and rat, but not hamster. Immunostainable material was also present in the epithelia of the vas deferens and seminal vesicles of mouse and rat, the only two species thus far examined. Since beta-EP was present in Leydig cells, we wondered whether this peptide could be identified in other steroid-producing tissues. When rat ovaries and adrenals were reacted with anti-beta endorphin, staining was demonstrated in corpus luteum and adrenal cortex. No staining was observed in the adrenal medulla or other portions of the ovary. In order to determine whether the beta-EP detected in the testis and epididymis was derived from a pituitary source, animals were hypophysectomized and tissues examined 2 weeks later. Both the Leydig cells and the epididymal epithelium remained immunostainable. In summary, immunostainable beta-EP has been identified in Leydig cells of five species. Stainable material is also present in the epithelium of other portions of the male reproductive tract and in steroid-secreting cells of the ovary and the adrenal. Such beta-EP may have a paracrine function in the testis and other portions of the male reproductive tract. PMID- 6291654 TI - Acetylcholine receptor kinetics. A description from single-channel currents at snake neuromuscular junctions. AB - Single-channel currents from acetylcholine receptor channels of garter snake neuromuscular junctions were recorded using the patch-clamp technique. Low concentrations of acetylcholine or carbamylcholine induced populations of single current events whose amplitudes and durations had unimodal distributions. The probability with which channel opening transitions occurred was time dependent, so that it was more probable for channels to open during the several hundred microseconds following a closing transition than during any later equivalent interval. The time-dependent distributions of duration and opening-transition probability were fitted by a sequential, reversible kinetic model in which the agonist binding steps occur before, and separately from, channel activation. This description allowed estimates to be obtained of both the opening (approximately 750s-1) and closing (approximately 500s-1) transition rates of these channels and of the mean lifetimes of the open- (approximately 2 ms) and the closed-channel state (approximately 200 mus) to which the open state was reversibly related. PMID- 6291655 TI - Single calcium-dependent potassium channels in clonal anterior pituitary cells. AB - Single Ca2+-dependent K+-channel currents were recorded in intact and excised inside-out membrane patches of the anterior pituitary clone AtT-20/D16-16. The frequency of channel openings and lifetimes depends both on membrane potential and on the Ca2+ concentrations at the inner membrane surface. The curve of the open-state probability of the channel as a function of membrane potential appears to translate along the voltage axis with changes in internal Ca2+ concentration. For Ca2+ concentrations between 10(-7) and 10(-6) M, the shift is consistent with the hypothesis that three Ca2+ ions are required to open a Ca2+-dependent K+ channel. Single channel conductances are estimated to be 124 pS in patches with normal external K+ (5.4 mM) and 208 pS in excised patches with symmetrical K+ (145 mM) across the membrane. Tetraethylammonium (20 mM) added to the cytoplasmic surface reversibly blocks the Ca2+-dependent K+ channel. PMID- 6291656 TI - Picosecond spectroscopy of Cu(II) cytochrome c. AB - We have observed a strong pH dependence in the relaxation rate of Cu(II) cytochrome c following excitation at 532 nm. At pH 8.0 the excited state relaxes with a lifetime of 10 +/- 5 ps while at pH extremes of 2.5 and 13.0 we find that the lifetime becomes longer than 1 ns. This change of more than two orders of magnitude in the lifetime may be due to the Cu coordination number, which is six at neutral pH but five at pH extremes. PMID- 6291657 TI - Unidirectional sodium and potassium fluxes through the sodium channel of squid giant axons. AB - Unidirectional 22Na-traced sodium influx or 42K-traced potassium efflux across the membranes of voltage-clamped squid giant axons was measured at various membrane potentials under bi-ionic conditions. Tetrodotoxin almost entirely eliminated the extra K+ efflux induced by short repetitive depolarizations in the presence of tetraethylammonium or 3,4-diaminopyridine. A method of determining the voltage dependence of the unidirectional flux through voltage-gated channels is described. This technique was used to obtain the unidirectional flux-voltage relation for the sodium channel in bi-ionic and single-ion conditions. It allows the determination of the unidirectional flux at the zero-current potential which, for influx, was found to be approximately 20% of the value measured 80 mV negative to the zero-current potential. The unidirectional flux ratio under bi ionic conditions was also measured and the flux ratio exponent found to average 1.15 with an external sodium and an internal potassium solution. A three-barrier, two-site, multi-occupancy model previously obtained for other conditions was found to predict a similar non-unity average for the flux ratio exponent. It is also shown that some single-occupancy models can predict non-unity values for the flux ratio exponent in bi-ionic conditions. PMID- 6291658 TI - An experimental study of the dielectric dispersion of solutions of cytochrome c at medium ionic strengths. AB - In this paper, the results are presented of measurements of the dielectric dispersions of horse heart cytochrome c molecules in various buffers. The data are fitted to the Cole-Cole relaxation model. The influence of the concentration and the ionic strength on the parameters that result from the Cole-Cole model is determined. The measured data are compared with calculations based on the model presented previously. Good agreement is found between the model and the observed data. PMID- 6291659 TI - Electrochemical reactions of horse heart cytochrome c at graphite electrodes. AB - The interaction of cytochrome c with a paraffin-wax-impregnated spectroscopic graphite electrode (WISGE) was studied in a medium consisting of 0.1 M potassium phosphate, pH 7.0, by means of differential pulse and cyclic voltammetry. Ferricytochrome c yields on voltammograms a single cathodic peak C around a potential of -0.3 V (vs. Ag/AgCl) and two anodic peaks AI and AII around the potentials of 0.66 and 0.89 V, respectively. Cathodic peak C corresponds to a catalytic reaction during which ferricytochrome c is reduced to ferrocytochrome c: ferricytochrome c is then regenerated by chemical oxidation of ferrocytochrome c by oxygen adsorbed at the WISGE surface. The first, more negative anodic peak AI corresponds to anodic electrochemical oxidation of tyrosine residues, whereas the second, more positive anodic peak (peak AII) corresponds to an anodic reaction of haemin. Voltammetry at a WISGE may provide a valuable technique for obtaining data about cytochrome c properties on electrically charged surface. PMID- 6291660 TI - A model for the calculation of the dielectric dispersion and the dipole moment of globular proteins in solution. AB - We describe a new procedure whereby the magnitude of the dielectric dispersion of a solution of globular protein molecules can be calculated. The protein molecule is considered to have spherical symmetry and the charged residues are thought to be situated in a medium whose dielectric constant increases continuously as a function of the distance from the centre of mass. The dipole moment of the protein in the solution is made up of two parts: the intrinsic dipole moment due to the charge distribution of the protein and the dipole moment due to polarization of the medium and the ionic cloud. When the model is applied to solutions of cytochrome c it is found that polarization of the medium results in a decrease in the dielectric dispersion amplitude. The mean square dipole moment calculated with the help of this method indicates that the fluctuation of the configurations cannot be responsible for the large dispersion in the megahertz region. PMID- 6291661 TI - Triiodothyronine induces an increase in cyclic GMP in bullfrog tadpole tissues. AB - Cyclic GMP (cGMP) and cyclic AMP (cAMP) were determined in bullfrog tadpole liver and tail fin using 125I-RIA. cGMP increased approx. 100% 1-6 h after the injection of T3 (3 X 10(-10) mol/g body wt.). Reducing the dose of T3 to 1, 3, and 10 X 10(-11) mol/g body wt. provided increases in cGMP of 50-100% above the control value after 2 h. In contrast, only small increases (less than 20%) in cAMP were observed 2-24 h after T3 injection. We conclude that T3 produces a rapid and significant increase in cGMP in the liver and tail fins of premetamorphic tadpoles. These results suggest that thyroid hormones in amphibia may not be an exclusively nucleus-mediated hormone. PMID- 6291662 TI - The cation pump as a switch mechanism controlling proliferation and differentiation in lymphocytes. PMID- 6291663 TI - Signalling mitogenesis in 3T3 cells: role of monovalent ion fluxes and cyclic nucleotides. AB - Quiescent 3T3 cells resting in the G0 phase of the cell cycle can be stimulated to reinitiate DNA synthesis by combinations of chemically diverse agents which act synergistically when added to cultures maintained in serum-free medium. Understanding of the mechanism(s) whereby extracellular agents interact in modulating cell proliferation requires the identification of the intracellular signals important for initiating a mitogenic response. Our recent evidence indicates that increases in ion fluxes involving Na+, H+, K+, and Ca2+ and elevations in cyclic AMP (cAMP) levels (produced by cholera toxin, adenosine agonists, prostaglandin E1, or cAMP derivatives) can act as mitogenic signals for Swiss 3T3 cells. We propose that initiation of DNA synthesis can be elicited by the synergistic interaction of two identifiable signals, namely, an enhanced rate of ion movements and an increase in the cellular level of cAMP. PMID- 6291664 TI - Inhibition of tumor growth by alkylation of the plasma membrane. PMID- 6291665 TI - Membrane changes during viral infection. AB - The effect of viruses on the surface membrane of susceptible cells during the entry and exit process has been studied. Haemolytic paramyxoviruses induce a non specific leakage to low-molecular-weight compounds during entry; other viruses do not show this effect. During exit, no such changes occur with any virus so far studied: some viruses are released without any obvious change at all in surface membrane function; in other cases, uptake of some nutrients is altered and there is a fall in membrane potential. These results do not support the hypothesis that a generalized membrane leakiness is a prerequisite for the synthesis and release of virus particles. PMID- 6291666 TI - Alterations in plasma-membrane functions after poliovirus infection. AB - After exposure of HeLa cells to poliovirus there is a rapid decline (within minutes) in fluorescence polarization of DPH (1,6-diphenyl-1,3,5-hexatriene). Within one hour after infection the (Na+/K+)ATPase activity of an isolated plasma membrane-rich fraction is enhanced, the cell volume decreases, and the intracellular concentration of a potent low-molecular-weight inhibitor of host protein synthesis increases. PMID- 6291667 TI - Membrane-mediated alterations of intracellular Na+ and K+ in lytic-virus-infected and retrovirus-transformed cells. AB - Infection of chick-embryo fibroblasts and other cells by certain animal viruses results in alterations in the intracellular concentrations of Na+ and K+. Dramatic alterations in monovalent-cation concentrations of lytic-virus-infected cells may favor the synthesis of viral proteins over cellular proteins. More subtle alterations in retrovirus-transformed cells may result in the expression of many morphological and biochemical changes associated with the transformed phenotype. PMID- 6291668 TI - Influence of serum and poliovirus infection on the transport and accumulation of AIB in HeLa cells. AB - The kinetics of active transport of the model amino acid AIB (aminoisobutyric acid) into HeLa cells were analysed. Serum withdrawal from cell-culture medium prior to transport assay strongly lowers the influx and steady-state distribution of AIB. Readdition of serum restores the rate of influx within 30 min, while the capacity to concentrate AIB is only partially reestablished. These changes are due to alterations in the activity of system A. In the time-course of poliovirus infection the initial rate of AIB transport shows two distinct changes. Early in the infection there is an increase in influx, followed by a strong decrease later in infection. These alterations are also attributed to transport system A. PMID- 6291669 TI - Topological and functional aspects of the proton conductor, F0, of the Escherichia coli ATP-synthase. AB - The isolated H+ conductor, F0, of the Escherichia coli ATP-synthase consists of three subunits, a, b, and c. H+-permeable liposomes can be reconstituted with F0 and lipids; addition of F1-ATPase reconstitutes a functional ATP-synthase. Mutants with altered or missing F0 subunits are defective in H+ conduction. Thus, all three subunits are necessary for the expression of H+ conduction. The subunits a and b contain binding sites for F1. Computer calculations, cross links, membrane-permeating photo-reactive labels, and proteases were used to develop tentative structural models for the individual F0 subunits. PMID- 6291670 TI - 3',5'-cyclic-nucleotide-dependent protein kinase of squamous cell carcinoma of the prostate. AB - An adenosine 3'5'-cyclic-monophosphate (Cyclic AMP)-dependent protein kinase has been identified and partially purified from the rat prostate tumor induced by 20 methylcholanthrene. This enzyme is stimulated 2- to 3-fold by the nucleotide. Equilibrium studies at pH 5.0 suggest the presence of a major class of binding site for cyclic AMP with an association constant of approximately 10(8) M-1. The concentration of binding site is about 1 pmol/mg of protein of the enzyme preparation. The enzyme is stimulated by other cyclic nucleotides as well, but only by higher concentrations. In comparing the ability of different histone subfractions, casein and protamine, to serve as substrate for this particular protein kinase, maximal cyclic-AMP-dependent enzyme activity was observed with histones. The results suggest that factors contributing to the malignant growth of the prostatic tissue do not directly involve changes in the characteristics of a cyclic-AMP-dependent protein kinase. PMID- 6291671 TI - The current status of NMR imaging. AB - A brief account is given of the current status of nuclear-magnetic-resonance (NMR) imaging for medical purposes. The procedures in present use for two- and three-dimensional NMR imaging are outlined and examples given. The quality of images approaches that of computerized tomography X-ray scans and demonstrates superior tissue discrimination and pathological discrimination. Guidelines for safe operation are discussed. Clinical trials of commercial NMR scanners at an advanced stage. PMID- 6291672 TI - Formation of pyrophosphate on hydroxyapatite with thioesters as condensing agents. AB - "Energy-rich" thioesters are shown to act as condensing agents in the formation of pyrophosphate on hydroxyapatite in the presence of water at ambient temperature. The yield of pyrophosphate based on thioester ranges from 2.5% to 11.4% and depends upon the pH and concentration of reactants. Reaction of 0.130 M hydroxyapatite suspended in a solution of 0.08 M sodium phosphate and 0.20 M imidazole hydrochloride (pH 7.0) with 0.10 M N,S-diacetylcysteamine for 6 days gives the highest yield of pyrophosphate (11.4%). Pyrophosphate formation requires the presence of hydroxyapatite, sodium phosphate and the thioester, N,S diacetylcysteamine. The related thioester, N,S-diacetylcysteine, also yields pyrophosphate in reactions on hydroxyapatite. PMID- 6291673 TI - Prebiotic adenine synthesis via HCN oligomerization in ice. AB - Adenine is produced (after hydrolysis) when 0.01 M solutions of HCN are adjusted to pH 9.2 with NH4OH and are frozen at -2 degrees C for 60-100 days. The addition of glycolonitrile (the cyanohydrin of formaldehyde) increases the yield of adenine under these conditions by about five-fold. These results confirm and extend an earlier suggestion that purine synthesis on the prebiotic Earth might have occurred in frozen, dilute solutions of HCN. PMID- 6291674 TI - A defect in the oxidative metabolism of human polymorphonuclear leukocytes that remain in circulation early in hemodialysis. AB - Human granulocytes harvested from uremic volunteers 15 min after the initiation of dialysis (at the nadir of neutropenia) were compared to predialysis controls. These intradialysis cells had a significant defect in peak luminol-enhanced chemiluminescence in response to opsonized zymosan, f-Met-Leu-Phe, and phorbol myristate acetate relative to predialysis control cells from the same patients. This defect could not be explained by a decrease in PMN myeloperoxidase concentration. H2O2 secretion by intradialysis cells (2 patients) was also depressed relative to predialysis controls. The ability to perform an independent function, orientation (polarization), was normal in both pre- and intradialysis cells relative to control. Whereas 125I-labeled formyl peptide binding studies demonstrated identical values for affinity and receptor number for predialysis and normal control cells, intradialysis cells displayed a 27% decrease in receptor number. This decrease in available receptor number. This decrease in available receptors may be related to the decreased chemiluminescence observed in response to f-Met-Leu-Phe. Furthermore, the results are consistent with the hypothesis that a defective PMN population remains in the circulation during the neutropenia of hemodialysis. PMID- 6291675 TI - Increased Ca++, Mg++, and Na+ + K+ ATPase activities in erythrocytes of sickle cell anemia. AB - To determine whether diminished activity of the Ca++ extrusion pump could account for the high levels of red blood cell (RBC) Ca++ in sickle cell anemia (SS), we measured calmodulin-sensitive Ca++ ATPase activity in normal and SS RBC. Hemolysates prepared with saponin were compared, since such preparations expressed maximum ATPase activities, exceeding isolated membranes or reconstituted systems of membranes plus cytosol, SS RBC hemolysates had greater Ca++ ATPase activity than normal hemolysates; they exhibited higher Mg++ and Na+ + K+ ATPase activities as well. Assays on density (age) fractions of SS and normal red cells demonstrated that all ATPase activities were highest in low density (young) cells, and activities in SS red cells exceeded those in normals in all fractions studied. Thus, when studied under conditions that maximize enzyme activity, Ca++ ATPase activity, like Mg++ and Na+ + K+ ATPase, is actually increased in SS RBC, probably due to the young red cell population present. The elevated Ca++ levels in these cells are more likely due to an increased Ca++ leak or abnormal calcium binding than to defective extrusion by the ATPase pump. PMID- 6291676 TI - The effect of deoxygenation on red cell density: significance for the pathophysiology of sickle cell anemia. PMID- 6291677 TI - The physical and chemical features of Cannabis plants grown in the United Kingdom of Great Britain and Northern Ireland from seeds of known origin. AB - Cannabis plants have been grown in the United Kingdom of Great Britain and Northern Ireland from seeds taken from seizures of cannabis of known geographical origin and chemistry. The gross physical appearance and cannabinoid patterns of many of the cannabis samples produced in the United Kingdom were closely related to those of the parents. However, some notable exceptions were recorded. There were wide variations in actual tetrahydrocannabinol content between plants grown from different seedstock and rather smaller variations within the groups grown from the same seedstock. Cannabis produced in the united Kingdom and higher tetrahydrocannabinolic acid/tetrahyrocannabinol ratios than imported material. PMID- 6291678 TI - Steroids in dermatology. PMID- 6291679 TI - Steroids in renal disease. PMID- 6291680 TI - Malignant fibrous histiocytoma: a recurring disease. PMID- 6291681 TI - Inflammatory fibrous histiocytoma. AB - The name inflammatory fibrous histiocytoma (FH) is applied to a rare group of disorders which is not well known to surgeons but which may require surgical treatment. The name is misleading in that although there may be features of inflammation, the clinical progress, absence of causative organism and fatal outcome seem to place it within the sphere of malignancy. The condition is fully described by Kyriakos and Kempson (1976) and a case with a periorbital lesion managed by multiple surgical excisions, the "berry-picking" technique, is documented by Jones et al., (1979). The average age is over 50 years and the sex distribution about equal. In this paper a patient much younger than average is presented to illustrate the problems of diagnosis and management. During the course of treatment an observation was made on the "take" of a free graft on the cortex of the sternum. PMID- 6291682 TI - Membrane properties and excitatory neuromuscular transmission in the smooth muscle of dog cerebral arteries. AB - 1 Drug actions on electrical and mechanical properties of smooth muscle cells and neuromuscular transmission in the canine cerebral arteries were investigated by use of microelectrode and isometric tension recording methods. 2 In the basilar and middle cerebral arteries, the resting membrane potentials were--49.4 mV and 51.7 mV, respectively, the length constants 0.57 mm and 0.45 mm, respectively and the time constants 142 ms and 118 ms, respectively. 3 Outward current pulses did not evoke the spike in either artery but did evoke the spike under conditions of pretreatment with 10 mM tetraethylammonium (TEA). 4 The maximum slope of depolarization produced by a ten fold increase in [K]o plotted on a log scale was 40.1 mV in the basilar artery and 42.2 mV in the middle cerebral artery. 2 Nicotinamidoethyl nitrate, the K-permeability accelerator, had no effect on the membrane potential. 5 K-free or ouabain [10(-5)M] treatment slightly depolarized the membrane. Re-addition of K [5.9 mM] hyperpolarized the membrane by several mV. Thus, the contribution of an active Na-K pump in the membrane potential seems to be small. 6 In both arteries, acetylcholine, adenosine, noradrenaline and isoprenaline in concentrations up to 10(-5)M did not modify the membrane potential and resistance, while 5-hydroxytryptamine (over 10(-8)M) and ATP (over 10(-5)M) depolarized the membrane, decreased the membrane resistance and produced a dose-dependent contraction. Adenosine suppressed the contraction evoked by excess [K]o (39.8 mM). 7 Perivascular nerve stimulation produced excitatory junction potentials (e.j.ps). Often e.j.ps were followed by a hyperpolarization. Repetitive stimulation produced facilitation after several stimuli and depression followed. In some cells, this depression appeared without facilitation. 8 The e.j.ps ceased with pretreatment with guanethidine (10(-6)M) or tetrodotoxin (3 X 10(-7)M), while phentolamine (10(-7)M) and yohimbine (10(-7)M) enhanced the amplitude of e.j.ps. ATP (10(-5)M) and noradrenaline (10(-6)M) suppressed and prazosin had little effect on the e.j.ps. Atropine (10(-6)M) also had no effect on the e.j.ps. 9 Specific features of the cerebral artery and systemic vascular beds were compared, and the features of adrenoceptors on the smooth muscle membrane were compared with findings in other vascular beds. PMID- 6291683 TI - Effects of ketamine and of high pressure on the responses to gamma-aminobutyric acid of the rat superior cervical ganglion in vitro. AB - 1 The method of Brown & Marsh (1974) for recording of surface potentials from the rat superior cervical ganglion has been adapted for use in a high pressure chamber in order to study the effects of high pressure of helium and the possible interactions with the effects of general anaesthetics. 2 Helium pressure of 130 atm did not alter the amplitude of the responses recorded from the ganglion in response to gamma-aminobutyric acid (GABA) application (9.7 and 19.4 microM) but the amplitude of responses to a nicotinic agonist were depressed. 3 Ketamine, at concentration between 18 and 180 microM, considerably potentiated the responses of the ganglion to GABA. 4 Helium pressure (130 atm) did not reverse the potentiation of GABA by ketamine. 5 The results are discussed in connection with the ability of ketamine to oppose the behavioural effects of high pressure. PMID- 6291684 TI - Tricyclic antidepressants vary in decreasing alpha 2-adrenoceptor sensitivity with chronic treatment: assessment with clonidine inhibition of acoustic startle. AB - 1 Clonidine inhibition of the acoustic startle reflex in the rat was used as a behavioural measure of alpha 2-adrenoceptor sensitivity following acute or chronic administration of tricyclic antidepressants. 2 Chronic (14 day) administration of desipramine (10 mg/kg, i.p.) attenuated the depressant effect of clonidine (20 or 40 microgram/kg) on the startle reflex. 3 No change in response to clonidine was obtained after chronic treatment with two other tricyclic antidepressants, amitriptyline (10 mg/kg) or iprindole (5 mg/kg). 4 Acute administration of these tricyclics (1 h) did not modify the effect of clonidine on startle. 5 It is suggested that the development of alpha 2 adrenoceptor subsensitivity produced by chronic tricyclics may be unique to those compounds, such as desipramine, which are active in blocking the uptake of noradrenaline. PMID- 6291685 TI - Stimulation of arachidonic acid metabolism and generation of thromboxane A2 by leukotrienes B4, C4 and D4 in guinea-pig lung in vitro. AB - 1 Leukotriene C4 (LTC4), LTD4, slow-reacting substance of anaphylaxis (SRS-A) (from guinea-pig lung), bradykinin (Bk) and arachidonic acid (AA) release thromboxane A2 (TxA2) and prostaglandin-like materials from guinea-pig isolated perfused lungs. 2 Release of TxA2 induced by LTC4 and LTD4 is inhibited by a thromboxane synthetase inhibitor, imidazole (2.9 mM). 3 Mepacrine (200 microM), a phospholipase inhibitor, inhibits release of TxA2 and prostaglandin-like materials caused by SRS-A and Bk but not that due to exogenous AA 4 Leukotrienes B4, C4 and D4 are approximately equipotent in inducing dose-related contractions of guinea-pig parenchymal strips (GPPs). 5 Leukotriene-induced contractions of GPPs are greatly inhibited by imidazole (2.9 mM), carboxyheptylimidazole (24 microM) and mepacrine (400 microM). 6 FPL 55712 (1.9 microM), the SRS-A antagonist, blocks contractions of GPPs induced by LTC4 and LTD4 but not those due to LTB4 or Bk. 7 Tachyphylaxis to LTB4 occurs in GPPs but not to LTC4 or LTD4. 8 These results suggest that in guinea-pig lung in vitro, LTB4, LTC4 and LTD4 activate a phospholipase with subsequent generation of cyclo-oxygenase products of which TxA2 plays an important role. PMID- 6291686 TI - The effects of monovalent and divalent cations on the alpha-adrenoceptor of intact human platelets. AB - 1 We have examined the effects of monovalent and divalent cations and purine nucleotides on the binding of agonists and antagonists to the alpha-adrenoceptor of intact human platelets. 2 Replacement of Na+ (150mM) by NH4+ (150mM) in the incubation medium significantly reduced the binding affinity of [3H] dihydroergocryptine (P less than 0.05) but did not alter the binding capacity. The competitive binding affinity of adrenaline and noradrenaline was unaltered. 3 The addition of Ca2+ (1mM) or Mg2+ (1mM) to the platelet suspension significantly reduced the platelet alpha-adrenoceptor capacity as indicated by either [3H] dihydroergocryptine (P less than 0.05) or [3H]-yohimbine (P less than 0.01; Ca2+ only). 4 The addition of Ca2+ (1mM) or Mg2+ (1mM) had no effect on the binding affinity of [3H]-dihydroergocryptine but significantly reduced that of [3H] yohimbine (P less than 0.05). The competitive affinity of adrenaline and noradrenaline determined by inhibition of [3H]-dihydroergocryptine binding, was unchanged in the presence of either cation. 5 Addition of the purine nucleotides ADP, ATP, GDP or GTP (final concentration 10 microM), either alone or in the presence of 1 mM Ca2+ or 1 mM Mg2+, had no effect on the binding of [3H] dihydroergocryptine or on the competitive affinity of adrenaline or noradrenaline. 6 We conclude that the alpha-adrenoceptor of intact human platelet displays the binding characteristics of the alpha 2L form of the receptor previously identified in the platelet lysate preparation. PMID- 6291687 TI - Dopamine acts at the same receptors as noradrenaline in the rat isolated vas deferens. AB - 1 The proposal that dopamine activates a different population of receptors from those activated by noradrenaline and phenylephrine to cause contraction of the rat vas deferens has been investigated using a preparation of the epididymal half of this tissue. 2 In preparations preincubated in cocaine, oestradiol and propranolol, to block sites of amine loss and beta-adrenoceptors, noradrenaline was the most, and dopamine the least, potent of the three agonists. Phentolamine competitively inhibited each of the agonists to a similar extent. Prazosin also inhibited the actions of the three agonists to a similar extent. These results indicate that the three agonists activate a single population of alpha 1 adrenoceptors to cause contraction in this preparation. 3 In experiments using the prostatic half of the rat vas deferens, in the presence of cocaine, oestradiol, propranolol and prazosin, noradrenaline was approximately 40 times more potent than dopamine in causing inhibition of twitches induced by electrical field stimulation. Yohimbine competitively antagonized the effects of the two agonists to a similar extent indicating that both act at the same population of alpha 2-adrenoceptors. 4 Taken together, these findings do not lend support to proposals that there are populations of specific dopamine receptors located pre- and postjunctionally in the rat vas deferens. PMID- 6291688 TI - Histamine H1-agonist potentiation of adenosine-stimulated cyclic AMP accumulation in slices of guinea-pig cerebral cortex: comparison of response and binding parameters. AB - 1 A range of histamine analogues have been examined as potentiators of the adenosine-stimulated accumulation of cyclic adenosine 3',5'-monophosphate (cyclic AMP) in slices of guinea-pig cerebral cortex. Dose-response curves were constructed for the 6 most active compounds and characterized in terms of the IC50, the slope and the maximum response attainable relative to that of histamine. 2 Histamine, 2-thiazolylethylamine and N alpha-methylhistamine produced a maximal or near maximal response. N alpha, N alpha-dimethylhistamine and 2-methylhistamine appear to be partial agonists. 3 The response to all the agonists was practically abolished by mepyramine 1 microM, indicating that the response is mediated largely or wholly via histamine H1-receptors. 4 The relative potencies of the agonists on cyclic AMP accumulation were in general similar to relative potencies in causing contraction of intestinal smooth muscle. The biggest difference was observed with N alpha-methylhistamine. 5 The histamine analogues were also examined as inhibitors of [3H]-mepyramine binding in homogenates of guinea-pig cerebral cortex. The inhibition curves were characterized in terms of IC50, the slope and the maximum percentage inhibition. This last value was compared with the inhibition produced by promethazine 2 microM. 6 For the 6 most potent agonists, the EC50 for cyclic AMP accumulation was compared with the IC50 against [3H]-mepyramine binding, corrected for inhibition of non-receptor binding and for competition with [3H]-mepyramine. With the possible exception of 2-pyridylethylamine, the values did not differ by more than a factor of 3. PMID- 6291689 TI - Frequency-dependent depression of ganglionic transmission by propranolol and diltiazem in the superior cervical ganglion of the guinea-pig. AB - 1 Effects of propranolol and diltiazem on ganglionic transmission in the superior cervical ganglion of the guinea-pig were investigated with intracellular recording techniques. 2 Propranolol and diltiazem (5 X 10(-6)-10(-5) M) induced a transmission failure in the ganglion upon preganglionic nerve stimulation at high frequency (25-30 Hz) without affecting action potentials induced by direct stimulation of the soma membrane, or potentials induced by iontophoretically applied acetylecholine. 3 The results suggest that propranolol and diltiazem may act on preganglionic nerve terminals to inhibit Ca2+ influx in a frequency dependent manner. These agents may depress excess sympathetic activity without much affecting normal ganglionic transmission. PMID- 6291690 TI - The use of low concentrations of divalent cations to demonstrate a role for N methyl-D-aspartate receptors in synaptic transmission in amphibian spinal cord. AB - 1 Synaptic potentials and the responses of frog spinal cord to various acidic amino acids were examined by means of the sucrose gap recording technique. 2 Divalent cations (50-250 microM) specifically antagonized responses evoked at N methyl-D-aspartate (NMDA) receptors by N-methyl D,L aspartic acid (NMDLA). The rank order of potency was Ni2+ greater than Co2+ greater than Mg2+ greater than Mn2+. Responses to glutamate and aspartate were relatively insensitive to these concentrations of divalent cations. 3 The rank order of potency for divalent ions (1 mM) for antagonism of synaptic transmission in bullfrog sympathetic ganglia was Mn2+ greater than Co2+ greater than Ni2+ greater than Mg2+. Thus synaptic transmission in ganglia was especially sensitive to Mn2+ whereas NMDLA responses were especially sensitive to Co2+ and Mg2+. 4 It was possible to depress selectively the dorsal root-dorsal root potential (DR-DRP) and dorsal root ventral root potential (DR-VRP) of frog spinal cord using low doses of Co2+ or Mg2+ which did not affect VR-DRP (ventral root-dorsal root potential). It was not possible to produce this selective depression of DR-DRP and DR-VRP with Mn2+, as this cation non-selectively depressed all responses. 5 These results suggest that: (i) divalent cations do not antagonize NMDLA responses by blocking Ca2+ channels which may mediate the response; (ii) postsynaptic NMDA receptors are activated by a neurotransmitter involved in the DR-DRP and DR-VRP pathways but not by any neurotransmitters involved in the VR-DRP pathway; (iii) the neurotransmitter activating NMDA receptors in amphibian spinal cord may be an aspartate-like substance rather than aspartate itself or glutamate. PMID- 6291691 TI - The effects of theophylline and 4-(3-butoxy-4-methoxybenzyl)-2-imidazolidinone (RO 20-1724) on protein secretion from rat parotid gland. AB - 1 The effects of two chemically distinct cyclic nucleotide phosphodiesterase (PDE) inhibitors on protein secretion from superfused rat parotid gland were studied.2 In the presence of 1.0 mM Ca(2+), Ro 20-1724 (10 muM), an imidazolidinone derivative, increased the secretory response to isoprenaline 100% and the isoprenaline-dependent accumulation of adenosine cyclic 3',5' monophosphate (cyclic AMP) 300-400%. At this concentration Ro 20-1724 alone did not cause protein secretion, accumulation of cyclic AMP or significantly inhibit PDE activity in cell-free preparations from parotid gland.3 In the absence of added Ca(2+) and in the presence of 1.0 mM EGTA, Ro 20-1724 inhibited the secretory response to isoprenaline 65% while increasing isoprenaline-dependent cyclic AMP accumulation 200%.4 In the presence of Ca(2+), theophylline (10 mM) stimulated protein secretion but did not cause the accumulation of cyclic AMP. When combined with isoprenaline the rate of secretion was greater than the sum of the effects of the individual drugs but there was no effect of theophylline on the isoprenaline-dependent accumulation of cyclic AMP.5 Theophylline-stimulated protein secretion is increased by omitting Ca(2+) from the superfusion medium without any detectable change in cyclic AMP accumulation. Under these conditions Ro 20-1724 inhibits theophylline-stimulated protein secretion and the maximum rate of protein secretion in the presence of isoprenaline and theophylline is no greater than that seen with either agent alone.6 It is concluded that the theophylline effects do not result from inhibition of PDE. It is suggested that the primary action of both drugs on parotid gland acinar cells is to alter the distribution of intracellular Ca(2+). Ro 20-1724 may also inhibit Ca(2+)/calmodulin activated enzymes such as PDE. PMID- 6291693 TI - Spectrum of the mu, delta- and kappa-binding sites in homogenates of rat brain. AB - 1 In homogenates of rat brain, the binding characteristics of tritiated opiates and opioid peptides were examined and the relative capacities of mu-, delta- and kappa-binding sites of the opiate receptor determined by saturation analysis.2 In competition experiments, binding of the selective mu-ligand [(3)H]-[D Ala(2),MePhe(4),Gly-ol(5)]enkephalin at the mu-site was displaced by [D-Ala(2),D Leu(5)]enkephalin with rather low affinity (K(I) = 12.6 nM) and more readily by the ketazocine-like compounds (-)-ethylketazocine (K(I) = 3.1 nM) and (-) bremazocine (K(I) = 0.32 nM), which also displaced the binding of [(3)H]-[D Ala(2),D-Leu(5)]enkephalin from the delta-site. In contrast, the binding to the kappa-site was easily displaced by ethylketazocine (1.0 nM) and bremazocine (0.37 nM) but not by the mu-ligand [D-Ala(2),MePhe(4),Gly-ol(5)]enkephalin (K(I) = 2000 3000 nM) or the delta-ligand [D-Ala(2),D-Leu(5)]enkephalin (K(I) > 20,000 nM).3 The dissociation equilibrium constant (K(D)) and the binding capacity (pmol/g) of the mu-binding site were determined with the selective mu-ligand [(3)H]-[D Ala(2),MePhe(4),Gly-ol(5)]enkephalin. For the delta-site, [(3)H]-[D-Ala(2),D Leu(5)]enkephalin was used in the presence of unlabelled [D-Ala(2),MePhe(4),Gly ol(5)]enkephalin in order to suppress cross-reactivity to the mu-binding site. For the estimation of kappa-binding, [(3)H]-(+/-)-ethylketazocine or [(3)H]-(-) bremazocine were used in the presence of unlabelled mu- and delta-ligands for the suppression of cross-reactivities to the mu- and delta-binding sites.4 In rat brain the capacity of the mu-binding site was 7.3 pmol/g brain, that of the delta binding site 6.7 pmol/g brain and that of the kappa-binding site 2.0 pmol/g brain. Thus, the kappa-binding site had the lowest value whereas in the guinea pig brain the capacity of the mu-binding site was lower than that of the delta- or kappa-binding site. PMID- 6291692 TI - Irreversible inhibition of epithelial sodium channels by ultraviolet irradiation. AB - 1 The effects of u.v. irradiation at 254 nm and 350 nm on sodium transport across frog skin epithelium have been investigated. 2 Irradiation at 254 nm but not at 350 nm produces a dose-dependent, functionally selective blockade of sodium transport. The effect is apparently due to the irreversible closure of apical sodium channels. 3 The amiloride-sensitive conductance was directly related to sodium transport as measured by short circuit current (SCC) both in normal and irradiated tissues, although both conductance and current were reduced in irradiated tissues. 4 The sensitivity of epithelia to irradiation at 254 nm was defined from the rate constants for the decline in SCC during three 2 min periods of irradiation at 1850 microW cm-2. The rate constant for the initial 2 min irradiation was 0.093 +/- 0.008 min-1. 5 Lowering the sodium concentration to 5.5 mM from 110 mM increased the rate constant to 0.141 +/- 0.014 min-1, consistent with the view that more functional sodium channels exist at lowered sodium concentration. 6 Lowering the temperature to 7 degrees C from 23 degrees C reduced the rate constant to 0.032 +/- 0.007 min-1 suggesting that blockade of channels is not due to a direct interaction with photons. 7 Using a variety of experimental protocols we were unable to demonstrate that bromamiloride or iodoamiloride can act as photoligands for sodium channels in the epithelium of Rana temporaria. This is in contrast to earlier reports with other epithelia. PMID- 6291694 TI - Magnesium deficiency and diuretics. PMID- 6291695 TI - Diuresis or urinary alkalinisation for salicylate poisoning? AB - Forty-four adults with aspirin poisoning were treated with oral fluids only, standard forced alkaline diuresis, forced diuresis alone, or sodium bicarbonate (alkali) alone. Alkali alone was at least as effective and possibly more effective than forced alkaline diuresis in enhancing salicylate removal. Unlike the diuresis regimens it did not cause fluid retention or biochemical disturbances. The renal excretion of salicylate depends much more on urine pH than flow rate, and forced diuresis alone had little useful effect. In overdosage aspirin causes sodium and fluid retention and may impair renal function. Attempts to force a diuresis are potentially hazardous and the spurious fall in plasma salicylate concentration caused by haemodilution gives a false impression of efficacy. Further studies are required to determine the optimum treatment for salicylate poisoning. PMID- 6291696 TI - Pop diets for weight reduction. PMID- 6291697 TI - Penicillinase-producing Neisseria gonorrhoeae in Great Britain, 1977-81. PMID- 6291699 TI - Metabolic consequences of salbutamol poisoning reversed by propranolol. PMID- 6291698 TI - Rate of inactivation of cytomegalovirus in raw banked milk during storage at -20 degrees C and pasteurisation. AB - Samples of milk from 23 mothers attending the department of obstetrics and gynaecology and 36 who donated milk to the department's milk bank were cultured for cytomegalovirus. Virus was isolated from samples from 12 of the milk donors but none of the mothers attending the department; follow-up studies during lactation in seven of these 12 women showed that five continued to excrete the virus. Samples were taken on three occasions from one woman who regularly excreted high titres of the virus. Storage at -20 degrees C for over three days reduced the titre by over 99%; after pasteurisation at 63 degrees C for eight minutes the milk did not contain any viable virus. It is recommended that raw banked milk used for feeding preterm babies should be kept frozen for at least 72 hours before feeding. PMID- 6291700 TI - Electroshock afterdischarges are related to neuronal oscillability: neuronal models of Aplysia. AB - Identifiable giant neurons of Aplysia explored intracellularly behave differently at the offset of an intracellular electroshock (IES) or after a synaptic 'tetanization', according to their functional type: neurons of the stable type depolarize and fire at the offset of the IES, anodal or cathodal, thus eliciting an afterdischarge (AD). The threshold of this AD is lowered if the neuron is destabilized, i.e. converted from the stable to an oscillatory type (for instance by decalcification). Neurons normally of the tonic type are more sensitive to an IES, eliciting a longer afterdischarge than the stable neurons. Extracellular electroshock (EES) anodal or cathodal, applied directly on desheathed somata of Helix give long-lasting afterdischarges at the offset. In addition, EES stimulating presynaptic terminals or axons leads to a high frequency synaptic input on remote neurons. At the offset of this input either prolonged synaptic afterdischarges or postsynaptic rebounds of the membrane potential sustaining bursts of decreasing amplitude denote apparent oscillatory properties of the synaptically activated neuron. Finally, any conversion by convulsants of tonic neurons to oscillators highly facilitates the elicitation of afterdischarges of axons simultaneous to paroxysmal depolarization shifts of the homologous somata. These results indicate that afterdischarge elicitation is highly facilitated (low threshold) in normal oscillatory neurons and/or chemically destabilized neurons. PMID- 6291701 TI - beta-Endorphin alters dopamine uptake by the dopamine neurons of the hypothalamus and striatum. AB - Opioid peptides have well-documented modulatory effects on the synaptic transmission of several neurotransmitters. In both hypothalamus and striatum there is dense innervation by the beta-endorphin and/or enkephalin neuronal systems, and physiologically relevant neuroregulatory interactions of these neurons occur with other important neurotransmitter neuronal systems, such as the dopaminergic tuberoinfundibular and nigroneostriatal systems. Previous reports have examined the effects of opioid peptides on release, synthesis and degradation of dopamine in these brain regions. In this report, we describe the effects of the intracerebral administration of beta-endorphin to increase dopamine (re)uptake by dopaminergic nerve terminals of the hypothalamus and striatum. The specific, high affinity uptake of [3H]dopamine by dopaminergic nerve terminals was studied in a synaptosomal preparation by pharmacological exclusion, using desmethylimipramine, of dopamine uptake into other monoaminergic nerve terminals. The augmentation of in vitro dopamine uptake in both hypothalamus and striatum following intracisternal administration of beta endorphin is specifically mediated by opiate receptors, since it could be prevented by pretreatment with an opiate receptor antagonist, naltrexone. In hypothalamus, the increased dopamine uptake represents a primary effect of beta endorphin on hypothalamic dopamine neurons and is not secondary to the opioid peptide-induced stimulation of prolactin secretion, since identical effects of beta-endorphin administration are seen in hypophysectomized and intact animals. The effect of beta-endorphin to increase hypothalamic dopamine uptake was not reflected by a change in the affinity constant for dopamine, but involved an increase in maximal initial velocity of uptake. Naltrexone blocked the effect of beta-endorphin to increase the Vmax for [3H]dopamine uptake by hypothalamic dopamine neurons in both intact and hypophysectomized rats. In vitro exposure of hypothalamic and striatal dopaminergic nerve terminals to a wide range of concentrations of beta-endorphin failed to reproduce the in vivo results; some concentrations of beta-endorphin produced small decreases in [3H]dopamine uptake which were not reversed by naloxone. The data of this study provide evidence for a further mechanism by which beta-endorphin may alter dopaminergic neurotransmission, namely by increasing dopamine reuptake into dopaminergic nerve endings. PMID- 6291702 TI - [3H]Ro5-4864 benzodiazepine binding in the kainate lesioned striatum and Huntington's diseased basal ganglia. PMID- 6291703 TI - Effects of naloxone on basal and stress-induced ACTH and corticosterone secretion in the male rat--site and mechanism of action. AB - The acute effects of naloxone upon basal and stress-induced secretion of ACTH and corticosterone (CS) in the adult male rat were investigated. Forty-five minutes subsequent to naloxone injection (5 mg/kg body wt, i.p.), basal serum levels of ACTH (by radioimmunoassay) and of CS (by corticosterone-binding globulin) were more than doubled, as compared to vehicle-treated animals. Upon exposure to either photic or audiogenic stress, the ACTH and CS secretory responses were greater in the naloxone-injected groups. In animals with complete hypothalamic deafferentation basal serum ACTH concentrations were significantly greater than in intact controls (2-fold), and naloxone elicited a further doubling of this parameter. In dexamethasone-pretreated rats (50 micrograms/animal, 4 h prior to naloxone), naloxone had no effect upon ACTH and CS secretion. This study demonstrates: (1) that acute naloxone administration leads to hypersecretion of ACTH, as well as of CS, in the adult male rat; and (2) that its effect is due to an action within the hypothalamo-hypophyseal unit. The data also suggest that these naloxone effects are not mediated by glucocorticoid hormones. PMID- 6291704 TI - Specific serotonergic projections to the lateral geniculate body from the lateral cell groups of the dorsal raphe nucleus. PMID- 6291705 TI - Light activation of dopaminergic neurons in rat retina is mediated through photoreceptors. PMID- 6291706 TI - Heterosynaptic interactions between septal and entorhinal inputs to the dentate gyrus: long-term potentiation effects. PMID- 6291707 TI - Evolution of striatal opiate receptors. PMID- 6291708 TI - Blockade of long-term potentiation in the dentate gyrus of freely moving rats by the glutamic acid antagonist GDEE. PMID- 6291709 TI - The effects of septal stimulation on spontaneous and tail-shock evoked neuronal activity in the brainstem of the rat. PMID- 6291710 TI - Sites and mechanisms of action of lidocaine upon the isolated spinal cord of the frog. AB - The sites of action of lidocaine on the responses evoked by stimulation of lateral column (LC) and dorsal root (DR) were studied in the isolated, intra arterially perfused spinal cord of the bullfrog. When the ventral root volley produced by stimulation was abolished by lidocaine, the presynaptic focal potential was almost unchanged. Intracellular recordings from motoneurons clearly demonstrated a marked reduction in amplitude of the EPSPs before the block of conduction of presynaptic fibers and the block of invasion of the neuron soma by antidromic spike potential. At low concentrations of lidocaine, the EPSPs elicited by LC stimulation produced shortening in time to peak, slowing in the decay time, decrease in amplitude and smaller changes in the later EPSPs of a train than the earlier ones. From the observations, it was concluded that the low concentrations of lidocaine affected primarily synaptic transmission in the spinal cord. The possible mechanisms of action of lidocaine were discussed. PMID- 6291711 TI - Innervation patterns of cutaneous hair receptors in cat. AB - Cat hair receptors were studied to determine whether they could be distinguished by the following receptive field characteristics: thickness of innervated guard hairs, distance between innervated follicles and receptive field size. Initially the receptors were classified as G1, GI, G2 or D on the basis of their velocity requirements for excitation, their degree of linear directionality, their vibrational sensitivity, and whether they were activated by movement of down hairs. It was found that the thickest guard hairs on the posterior aspect of a cat's hindleg were usually 4-5 times thicker than the thinnest guard hairs from the same area and that G1, GI and G2 neurons innervated the full range of guard hair thicknesses available. Although there was a tendency for thicker guard hairs to be more heavily innervated, none of the neurons studied innervated thick guard hairs exclusively. While movement of the down hair and most guard hairs within D mechanoreceptive fields easily evoked activity, a few guard hairs were regularly found for which mechanical displacement did not elicit a discharge even though they were well within the receptive field. Receptive field sizes and nearest neighbor distances between innervated follicles were smaller for D than for G1, GI and G2 receptors and greater for G1 than GI and G2 receptors. PMID- 6291712 TI - Enkephalin inhibition of inhibitory input to CA1 and CA3 pyramidal neurons in the hippocampus. AB - Enkephalin-induced excitation in the hippocampus has been attributed to the attenuation of inhibitory input as well as to augmentation of excitatory input to pyramidal neurons. We have further examined these possible mechanisms of enkephalin action, as well as the possibility that enkephalins may be affecting intrinsic membrane properties, by recording intracellularly from CA1 and CA3 pyramidal cells in the guinea pig hippocampal brain slice preparation. It was observed that the inhibitory synaptic potential was significantly decreased in the presence of leucine enkephalin and D-alanine, D-leucine-enkephalin (DADL), whereas the excitatory synaptic potential, revealed by block of the inhibitory postsynaptic potential (IPSP) by bicuculline, was unaltered. In addition, the response of pyramidal cells to pressure-applied GABA was unaffected by enkephalin, as were the voltage-dependent membrane conductances. The increase in excitability which was observed in both field potential and intracellular recordings to drop application of DADL must, then, be due to a purely presynaptic block of inhibitory interneurons in both the CA1 and CA3 areas of the hippocampus. PMID- 6291713 TI - Function of serotonin in physiologic secretion of growth hormone and prolactin: action of 5,7-dihydroxytryptamine, fenfluramine and p-chlorophenylalanine. AB - Growth hormone (GH) and prolactin secretory patterns in male rats chronically implanted with venous cannulae were studied in response to treatments known to acutely or chronically deplete central serotonin (5-HT) or to permanently destroy a high proportion of 5-HT neural structures. Spontaneous prolactin secretion was little affected by chronic 5-HT depletion produced by 5,7-dihydroxytryptamine (5,7-DHT) or parachlorophenylalanine (PCPA). Acute 5-HT release induced by fenfluramine produced sustained prolactin secretion in intact and 5-HT-depleted rats. GH secretion was inhibited by PCPA, and for 3 days following 5,7-DHT, but was normal by 7 and 21 days. Acute 5-HT release stimulated by fenfluramine unexpectedly inhibited GH in intact and 5-HT-depleted rats. It is concluded that 5-HT is a potent facilitatory neurotransmitter for prolactin; and it appears to have both facilitatory and inhibitory effects on GH. Recovery of GH secretory patterns within 7 days of 5,7-DHT treatment indicates that mechanisms act rapidly to compensate for depletion of 5-HT, and that it is difficult to achieve functional depletion of 5-HT for more than a few days. PMID- 6291714 TI - Changes in multiunit activity of nigral neurons induced by cholinergic and dopaminergic stimulation of the caudate nucleus. AB - The effects of stimulation of the caudate cholinergic and dopaminergic receptors on multiunit activity in the ipsilateral substantia nigra were studied in cats locally anesthetized, paralyzed and artificially respired. Cholinergic stimulation by intracaudate microinjections of 10 micrograms of carbachol diminished multiunit activity by 36% in the ventral substantia nigra (SN) and increased activity by 48% in the dorsal SN. This effect was abolished after electrolytic lesion of the ipsilateral striatonigral pathway. Opposite responses were observed following intracaudate administration of 20 micrograms of dopamine or 20 micrograms of D-amphetamine. Multiunit activity in the ventral SN increased by 36% and 34%, respectively, while the activity in the dorsal SN was reduced by 56% and 53%, respectively. Similar results were obtained in response to systemic administration of D-amphetamine. Extracaudate microinjections of carbachol and dopamine left multiunit activity in the SN unaffected. In conclusion, our results indicate an opposite action of caudate cholinergic and dopaminergic receptors on multiunit activity in the SN of the cat. PMID- 6291715 TI - Cholinergic excitation of mammalian hippocampal pyramidal cells. AB - Responses of CA1 pyramidal neurons to ACh were recorded with intracellular microelectrodes utilizing the in vitro guinea pig hippocampal slice preparation. ACh was delivered by drop or iontophoretic application to stratum oriens or stratum radiatum. Threshold dose for drop application was 1 mM. An initial hyperpolarization of 3.1 +/- 1.8 (S.D.) mV associated with a decrease in membrane input resistance (RN) of 21 +/- 9% (S.D.) occurred in about half the cells. This result is consistent with a presynaptic action of ACh mediated through excitation of inhibitory interneurons. This interpretation was supported by recordings of cholinergic excitatory responses from presumed interneurons, and repetitive spontaneous IPSPs from pyramidal neurons during the hyperpolarization. ACh evoked a slow depolarization (14.3 +/- 10.8 (S.D.) mV) accompanied by a peak increase in apparent input resistance (Ra) of about 60% in the majority of cells. Large increases in spike frequency were associated with these events but action potential shape was unchanged. Plots of Ra versus membrane potential following ACh application revealed that Ra increases were proportionately higher at depolarized membrane potential levels (less than or equal to -70 mV) in some neurons. In these cells Ra was increased significantly at -60 mV (28%), but only 6% at -75 mV. These results are consistent with the conclusion that ACh reduces a voltage-dependent gK, distinct from delayed rectification. ACh also induced a non voltage-dependent increase in Ra in some cells. ACh-evoked changes in Ra were long-lasting and gave rise to alterations in firing mode, with development of burst generation. ACh also transiently blocked after hyperpolarizations which followed spike trains in pyramidal neurons and presumed interneurons, an action which may be related to effects on a Ca2+-activated gK. PMID- 6291716 TI - Ionic mechanisms of cholinergic excitation in mammalian hippocampal pyramidal cells. AB - Intracellular recordings from CA1 hippocampal pyramidal neurons were obtained using the in vitro hippocampal slice preparation. Responses to ACh were monitored in the presence of blockers of voltage-dependent conductances including Mn2+, TTX and Ba2+. When Mn2+ was used to block voltage-dependent Ca conductance and possible indirect presynaptic cholinergic actions, ACh still induced a significant voltage-sensitive increase in apparent input resistance (Ra) (29%), but only an insignificant depolarization of membrane potential (Vm). When both voltage-dependent Ca and Na conductances were blocked by application of Mn2+ and TTX, respectively, ACh produced voltage-dependent increases in Ra (31%) without significant depolarization. In solutions containing TTX alone, ACh produced voltage-sensitive increases in Ra (32%) as well as a significant depolarization (6.2 +/- 3.1 mV (S.D.)). ACh transiently blocked the conductance increase which followed presumed Ca spikes, suggesting an action on the Ca-activated K-dependent conductance. The effects of Ba2+ application (100-200 microM) on Ra mimicked those of ACh. When ACh was applied to neurons in the presence of Ba2+, Ra remained unchanged, although Vm depolarization of 5-15 mV was still seen. The data indicate that ACh decreases both a voltage-dependent K conductance (distinct from that of the delayed rectifier) and a Ca-activated K conductance. Muscarinic cholinergic depolarization occurs as a result of blockade of K conductance, and is mediated by voltage-dependent Ca and Na conductances, and perhaps by presynaptic actions. PMID- 6291717 TI - [3H]-etorphine and [3H]-diprenorphine receptor binding in vitro and in vivo: differential effect of Na+ and guanylyl imidodiphosphate. PMID- 6291719 TI - Microsomal opiate receptors differ from synaptic membrane receptors in proteolytic sensitivity. AB - We have found that opiate receptors in smooth microsomal fractions differ from synaptic membrane-associated receptors in proteolytic sensitivity. With 3 proteases of different substrate specificities (trypsin, chymotrypsin and S. griseus protease) smooth microsomal opiate receptors from rat brain were consistently less sensitive to limited proteolysis than were synaptic membrane receptors. Thiamine pyrophosphatase, a luminal Golgi membrane marker enzyme, exhibited a similar resistance to S. griseus protease in microsomal preparations, while microsomal Na+/K+-ATPase (ouabain-sensitive) was readily destroyed by trypsin. We also discovered that smooth microsomal opiate receptors co-migrate with both Golgi membrane and endoplasmic reticulum marker proteins on equilibrium density gradients under isopycnic conditions. Electron microscopic examination of the Golgi-enriched fraction showed the typical cisternae frequently associated with isolated Golgi membranes. Synaptic junctions, presynaptic membranes, myelin and mitochondria were conspicuously absent from this fraction. Since the microsomes isolated in vitro showed similar topography to those in vivo, the binding sites for opiates could be localized on the luminal surface membranes of the microsomal fractions. The exquisite sensitivity of synaptic membrane opiate receptors to proteolysis suggests that these receptors are found on the extracellular surface of the synaptic junction. PMID- 6291718 TI - Use of the patch electrode for sensitive high resolution extracellular recording. PMID- 6291720 TI - GFAP immunoreactivity reveals astrogliosis in females heterozygous for jimpy. AB - The jimpy gene is a sex-linked recessive mutation which produces severe hypomyelination throughout the central nervous system (CNS) in affected male mice. The female carrier also expresses the mutation, but the degree of hypomyelination varies considerably among the tracts. In the optic nerve, patches of unmyelinated tissue are interspersed with myelinated zones; in the brain, myelination is retarded during development but recovers in the adult. We have previously shown, in the male mutant, that an astroglial hypertrophy is associated with the white matter. The present study was undertaken to determine the existence and extent of astrogliosis in the female carriers. In this immunocytochemical investigation using antiserum to glial fibrillary acidic protein (GFAP), we show that the optic nerve of the female carriers exhibits patches of gliosis which are similar in appearance and intensity to those found in affected males. These patches are not present in the white matter of the carriers' brain and spinal cord, but GFAP-immunoreactivity is more intense than in control females. Throughout certain fiber tracts (e.g. optic tract and internal capsule), the number of immunostained astrocytes is increased in comparison to controls; they are larger, have more processes, and are more intensely immunoreactive. The results of the present study show that the retardation of myelin formation is accompanied by an astroglial response and that the intensity of the gliosis closely parallels the extent of the hypomyelination. PMID- 6291721 TI - Sodium valproate enhances responses to GABA receptor activation only at high concentrations. AB - Sodium valproate (VPA) enhanced muscimol-induced depolarizations of rat cuneate afferent fibres, but only at millimolar concentrations. VPA (10 mM) also reduced the potency of picrotoxin as an antagonist of muscimol without affecting that of bicuculline. These findings are discussed in relation to the anticonvulsant properties and potency of VPA. PMID- 6291722 TI - An electrophysiological analysis of oxygen and pressure on synaptic transmission. AB - The effect of oxygen at high pressure (OHP), helium at 150 PSIG and 100% oxygen at ambient pressure on excitatory synaptic transmission was studied using the lobster walking leg neuromuscular preparation. Both 100% oxygen at 150 PSIG (7135 mm Hg oxygen) and helium at 150 PSIG (7000 mm Hg helium plus 135 mm Hg oxygen) produced a significant decrease in the amplitude of the junction potential (Vejp). The decrease in Vejp induced by OHP, however, was greater than with pressure alone. OHP also produced a significant decrease in short term facilitation. Exposure to 100% oxygen at ambient pressure produced a transient increase in Vejp and a large increase in frequency of miniature junction potentials. In each case the change in Vejp was due to changes in presynaptic release of transmitter since quantal content per fiber (M') was shown to decrease for OHP and helium at 150 PSIG and to transiently rise with 100% oxygen at ambient pressure. In addition, the response to exogenously applied glutamate (the putative neurotransmitter) was not affected by OHP, 150 PSIG helium or 100% oxygen at ambient pressure. This further indicates a presynaptic site of action. PMID- 6291723 TI - Immunohistochemical localization of 2', 3'-cyclic nucleotide 3'-phosphodiesterase in the retina. PMID- 6291724 TI - Variation in the size of synaptic contacts along developing and mature motor terminal branches. AB - The secretion of a quantum from groups of release sites (me) declines along the length of terminal branches at the amphibian neuromuscular junction. The morphological basis of this decline in me has been studied at neuromuscular junctions in juvenile muscles (fibre length 4 mm) and adult muscles (fibre length 22 mm). Serial sections cut through the length of the junctions have been examined with both light and electron microscopy. Juvenile junctions consist of two short (less than 50 micron) terminal branches; adult junctions often consist of 4 long (100-500 micron) terminal branches. Synaptic contacts are largest near the origins of terminal branches and decline in size towards the end of branches. The number of horseradish peroxidase-labelled synaptic vesicles at release sites, following stimulation in the presence of the enzyme, is largest for sites closest to the origin of the terminal branches. The results suggest that the decline in me along the length of terminal branches is due to decline in the size of release sites. PMID- 6291725 TI - Blockade of ganglionic transmission during synaptogenesis decreases alpha bungarotoxin binding in the chick ciliary ganglion and iris. AB - The role of normal synaptic activity in the biochemical development of the nervous system has been examined in the chick embryo. Chlorisondamine, a ganglionic blocking drug, was administered in ovo during the period of synaptogenesis in the parasympathetic ciliary ganglion. Following treatment with chlorisondamine, nicotinic binding sites (as measured with [125I] alpha bungarotoxin) were significantly reduced in both the ganglion and its end organ, the striated iris muscle. While the number of [125I] alpha-bungarotoxin binding sites eventually approached control levels in the iris, binding in the ciliary ganglion remained below normal values through hatching. PMID- 6291726 TI - Cerebellar macroneurons in microexplant cell culture. Methodology, basic electrophysiology, and morphology after horseradish peroxidase injection. AB - Cerebellar macroneurons, including Purkinje cells, survive and differentiate in long-term monolayer cultures, which are prepared by a partial dissociation procedure we refer to as a microexplant technique. Intracellular recording demonstrated that these neurons were functional, showing spontaneous spiking activity and electrical excitability, and both spontaneous and evoked synaptic activity. In order to further characterize cell types, light and electron microscopic studies were performed after intracellular iontophoresis of horseradish peroxidase. Purkinje neurons were identified by their form of dendritic arborization and numerous dendritic spines. Cortical granule cells and macroneurons derived from the deep nuclei could also be demonstrated. PMID- 6291727 TI - [Levels of streptococcal antibodies in acute infections and their sequelae]. PMID- 6291728 TI - Reduction of Salmonella typhimurium in laboratory-inoculated chickens by the use of erythrosine. PMID- 6291729 TI - Effect of streptococcal cell wall components on bone metabolism in vitro. AB - Cell was components from Streptococcus mutans NCTC 10449 and Streptococcus sanguis ATCC 10558 stimulated the release of 45Ca from prelabeled mouse calvariae in organ culture. Bone resorbing activity was not blocked by fetal calf serum. It was, however, blocked by calcitonin, an inhibitor of osteoclast-mediated bone resorption. Indomethacin, a prostaglandin synthetase inhibitor, partially blocked endogenous but not antigen-stimulated 45Ca, suggesting that antigen-stimulated bone resorption was not mediated by prostaglandins. The antigen preparations also had an inhibitory effect on the incorporation of 3H-proline and 3H-thymidine into explants of rabbit and rat calvariae, respectively. The inhibitory effect of antigen on 3H-proline incorporation was not altered by the presence of calcitonin, which suggests that it represented a real inhibition of protein synthesis and not a reflection that the bones were resorbing. These findings indicate that plaque bacterial antigens may contribute directly to the progressive loss of alveolar bone during periodontal disease. The assumption that only Gram-negative organisms play an important role In the etiology of periodontal disease appears incorrect. PMID- 6291730 TI - Effects of two bacterial products, muramyl dipeptide and endotoxin, on bone resorption in organ culture. AB - We have compared two components of bacterial cell walls, muramyl dipeptide (MDP) and lipopolysaccharide (LPS), for their effects on bone resorption as measured by the release of previously incorporated 45Ca. MDP is the smallest active component of peptidoglycan, whereas LPS is the active component of endotoxin. Fetal rat long bones were cultured for 5 days in a chemically defined medium supplemented with bovine serum albumin (BSA) or serum. LPS increased 45Ca release at concentrations of 0.03-1.0 microgram/ml. LPS further purified by electrolytic dialysis (ED-LPS) was active at 0.01 microgram/ml. ED-LPS was ineffective at such low concentrations in the presence of serum. The response to MDP was more variable than that to LPS, but bone resorption was stimulated at concentrations of 10(-7)-10(-5) M. MDP was less effective or inactive in medium supplemented with serum. Stereoisomers of MDP that do not have adjuvant activity caused minimal stimulation of bone resorption, whereas 6-0-steroyl MDP stimulated resorption at 10(-8) M. The stimulation of bone resorption by LPS and MDP was not inhibited by indomethacin. Both LPS and MDP increased lysosomal enzyme release in proportion to their effects on 45Ca release. LPS also markedly increased collagenase activity in the medium, but MDP did not. These results indicate that chemically different products of bacterial cell walls can stimulate bone resorption in vitro. These products may be distinguished by differences in dose response curve, serum inhibition, and collagenase release. PMID- 6291731 TI - Electron microscopy of avian osteopetrosis induced by retrovirus MAV.2-O. AB - Diaphyseal tibial bone of 12.5-13-day and 19-day-old embryos and 20-day-old hatched chicks infected with retrovirus MAV.2-O were examined by transmission electron microscopy. The viruses were associated with lining osteoblasts and osteocytes. Whereas the infection of the osteoblast layer seemed to be a transient stage, virus association with osteocytes was a constant and main ultrastructural feature. The viruses were found either in the osteoid or in the periosteocytic space of the bone lacunae. They arose from dense cytoplasmic areas located near the cell plasmalemma via a budding process. The newly budded virus particles often had a large tail or a fine stalk-like process lost in the extracellular space. The viruses underwent calcification by deposition of inorganic material and were incorporated in the bone trabeculae. No production of virus was observed in typical osteoclasts with well-differentiated ruffled borders. The viral-induced avian osteopetrosis seemed to result from increased bone deposition through stimulation of osteoblast and osteocyte activities, whereas osteoclastic bone resorption seemed to be undisturbed. PMID- 6291733 TI - Effects of medium composition on cell pigmentation, cytochrome content, and ferric iron reduction in a Pseudomonas sp. isolated from crude oil. AB - Cells of a pseudomonad associated with pipeline corrosion grown on a complex medium were orange in color and vigorously reduced ferric iron. The intensity of orange color of cells grown on a synthetic medium and their ability to reduce ferric iron was directly related to the iron content of the medium. Absorption spectrophotometric data show a direct relationship between color of cells, cytochrome content, and ability to reduce ferric iron. Carbon monoxide markedly, but not completely, inhibits the reduction of ferric iron. The data presented indicate that ferric iron can serve as a terminal electron acceptor for cytochrome-associated respiratory processes of this corrosive pseudomonad. PMID- 6291734 TI - Response of Clostridium perfringens and its L form to bacteriocins of C. perfringens. AB - Clostridium perfringens strain No. 28 and its penicillin-induced stable L form were treated with 10 different bacteriocins of C. perfringens. Viable count and labelled amino acid incorporation experiments revealed that the L form was sensitive to two and possibly three bacteriocins to which the bacillus was not, while both forms were commonly sensitive to two other bacteriocins and resistant to five others. Adsorption of bacteriocin, immunity factors, or perhaps uptake of bacteriocin might be proposed to explain the responses of these organisms to bacteriocins. PMID- 6291732 TI - Abnormal skeletal response to parathyroid hormone in dogs with chronic uremia. AB - The release of cyclic AMP from bone in response to stimulation with PTH 1-34 was examined in 20 dogs with long-term chronic renal failure (CRF) produced by unilateral nephrectomy and contralateral partial renal artery ligation. After 9 to 15 months of uremia, the tibiae were removed and perfused in vitro. Seven dogs with CRF served as controls, 7 dogs with CRF were treated with 24,25(OH)2D3 - 2.5 micrograms per day, and 6 CRF dogs underwent thyroparathyroidectomy (TPTX) 42 h before they were sacrificed. The release of cyclic AMP from bone in response to PTH 1-34 in the CRF dogs was severely reduced compared to the response observed in 7 dogs with normal renal function (net accumulation of cyclic AMP release 86 +/- 8.5 versus 426 +/- 59.0 pmol/30 min). Long-term treatment of uremic dogs with 24,25(OH)2D3 had no effect on the release of cyclic AMP by bone. However, the release of cyclic AMP was restored to normal levels in the CRF dogs that underwent thyroparathyroidectomy. All CRF dogs had secondary hyperparathyroidism and the fact that TPTX returned the cyclic AMP response to normal values suggests that desensitization to PTH of the adenylate cyclase system of bone exists in chronic uremia. PMID- 6291735 TI - Use of vitallium prosthesis to relieve jaundice in patients with obstruction at the bifurcation of the hepatic duct. AB - Fourteen patients with hepatic duct obstruction were treated by inserting a Vitallium prosthesis through the strictured area. Eleven had proven cholangiocarcinoma; in 3 a definitive diagnosis wsa not obtained. Jaundice was relieved and patients were afforded good palliation. Obstruction of the prosthesis has not been a problem and the absence of external drainage tubes has been a factor in improving the quality of life of these patients. The simplicity of the operation is emphasized. PMID- 6291737 TI - Hyperlucent lung: long-term complication of adenovirus type 7 pneumonia. PMID- 6291738 TI - Muscular pseudotumor of the breast following doxorubicin and radiation therapy for oat cell carcinoma of the lung. AB - Two male patients developed muscular pseudotumor of the breast following combined treatment of radiation and chemotherapy with cyclophosphamide, doxorubicin, methotrexate and procarbazine for oat cell carcinoma of the lung. The pathologic findings of the biopsy specimens revealed muscle and capillary changes similar to previously reported myocardiotoxicity from doxorubicin and radiation therapy. Discussed is a possible additive or synergistic toxic effect of doxorubicin and radiation therapy in the development of muscular pseudotumor of the breast. PMID- 6291736 TI - [Infections of the central nervous system caused by enterovirus: 223 cases seen at a pediatric hospital between 1973 and 1981]. AB - Between 1973 and 1981, 223 patients were seen at hopital Sainte-Justine in Montreal for enteroviral infection of the nervous system. In 161 the cause was documented by isolation of an enterovirus from the cerebrospinal fluid (CSF). The viruses most frequently isolated were echovirus 11 (36 isolates), echovirus 30 (24), coxsackievirus B2 (23), coxsackievirus B3 (19), echovirus 6 (18), coxsackievirus B5 (16), coxsackievirus A9 (15), echovirus 9 (13), echovirus 7 (12) and coxsackievirus B1 (11). Aseptic meningitis was diagnosed in 200 cases and encephalitis in 12. The remaining 11 patients presented with the features of septicemia or with convulsions. In 33 patients an enterovirus was isolated from the CSF in the absence of pleocytosis. Polymorphonuclear cell predominance was noted in the initial CSF sample in 95 cases; it was persistent in 11. There were five mixed infections; in three cases two viruses were isolated from the same CSF sample. Two patients died: one, a child with hypogammaglobulinemia, had fatal polioencephalitis; the other, a 6-month-old infant brought to the emergency room in unexplained cardiopulmonary arrest, had echovirus 6 meningitis. Of the 172 patients admitted to hospital 96 received parenteral antibiotic therapy. The impact of enteroviral infections of the central nervous system on hospital resources could be substantially reduced if a rapid, sensitive and specific laboratory method of diagnosing these infections were available. PMID- 6291740 TI - Krukenberg tumors of the ovary: a clinicopathologic analysis of 27 cases. AB - A series of 27 typical Krukenberg tumors of the ovary were analyzed. By definition, all examples were characterized by the presence of mucinous signet ring carcinoma cells within a cellular, nonneoplastic ovarian stroma. The patients' ages ranged from 20-70 years; almost one-half were 40 years of age or younger. A primary carcinoma of the stomach (16 cases) or colon (four cases) was found in 20 (90.9%) of 22 patients with available follow-up data. The primary gastrointestinal carcinomas had been diagnosed before emergence of the ovarian tumors in only five cases. The ovarian and gastrointestinal tumors were synchronously diagnosed in ten cases, while in five instances the primary carcinomas were not discovered until after the ovarian tumors had been treated. An acceptable primary extraovarian cancer was not detected in two women. Both had bilateral Krukenberg tumors and died with widespread carcinomatosis less than two years postoperatively. Typically, the ovarian tumors were bilateral, asymmetrically large and solid. Important histologic features included a greater abundance of intracellular neutral glycoproteins than acidic mucins, a storiform pattern of hyperplastic cortical stromal cells (44.4%) and carcinomatous emboli (51.6%). While the entity of "primary" Krukenberg tumor cannot be unequivocally denied, all women with typical Krukenberg tumors should be considered as having metastatic carcinoma, usually from the stomach, until proven otherwise. PMID- 6291739 TI - Malignant astrocytoma: hyperfractionated and standard radiotherapy with chemotherapy in a randomized prospective clinical trial. AB - A prospective randomized trial of 157 patients with malignant astrocytoma (Grade III or IV) was carried out at a single institution. The minimization technique ensured balanced distribution of prognostic factors between the treatment groups. All received oral lomustine (CCNU, 80 mg/m2) six weekly and hydroxyurea (HU, 3.5 gm/m2 over 5 days) three weekly, for one year or until recurrence, with doses adjusted for myelosuppression. Patients were randomized to daily (5000 rad in 25 fractions (fr) in 5 weeks) or Q3h (every 3 hours) Cobalt 60 irradiation (3600 4000 rad in 36-40 fr of 100 rad each, given 4 fr per day at 3-hour intervals over two weeks) Steroid therapy (up to 16 mg day dexamethasone) was permitted. Complications were moderate and equivalent in the two groups. No significant survival or toxicity differences were seen between the two groups. Age, initial performance status, and extent of surgical resection were found to be significant (P less than 0.01) prognostic factors for survival. Median survival of the whole group was 48 weeks with a minimum follow-up of one year. There was no advantage to large radiation fields. The hyperfractionation and daily regimens had similar efficacy and toxicity. Hyperfractionation with chemotherapy offers a useful alternative approach in the management of this disease. PMID- 6291742 TI - Mucinous carcinomas of the colon and rectum and their relation to polyps. AB - A study of 44 mucinous carcinomas (MC) from a series of 324 colorectal cancers was made (221 surgical resections and 103 endoscopic biopsies). This study showed that MC were associated, in a significantly higher proportion when compared to non MC, with polypoid adenomas of different kinds (hyperplastic polyps not included), in other segments of the surgical specimen (P less than 0.001). MC originated from adenomas, particularly villous, but also mixed and tubular, in a significantly higher proportion than non-MC (P less than 0.001). Carcinomas arising from adenomas were mucinous in 11/14 cases. The type of adenomas from which MC arose were characterized by usually having areas with a particular arborizing mucus hyperplasia. At the time of resection, MC had metastases (Stages C and D) more frequently then non MC (P less than 0.02). PMID- 6291741 TI - Biological markers and small cell carcinoma of the lung: a clinical evaluation of urinary ribonucleosides. AB - Five minor base ribonucleosides, primarily degradation products of transfer ribonucleic acid (tRNA), were evaluated as potential biological markers for patients with small cell carcinoma of the lung. The urinary concentration for pseudouridine, 1-methyladenosine, 1-methylinosine, N2-methylguanosine, and N2,N2 dimethylguanosine was determined by means of reversed-phase high performance liquid chromatography and quantitatively expressed as a function of creatinine excretion. Comparisons were made with carcinoembryonic antigen (CEA) plasma levels. The total frequency of elevated values for the five nucleosides in pretreatment urine samples was directly related to stage of disease with 24/60 (40%) determinations increased in 12 patients with limited disease and 69/85 (81%) in 17 patients with extensive disease. For these same patients, CEA levels were elevated respectively in 2/11 (18%) of the former and 9/17 (53%) of the latter group. The frequency and degree of elevation of the nucleoside/creatinine ratios in pretreatment samples from patients with extensive disease was correlated directly with increasing number of metastatic sites. Of the five nucleosides, the mean number elevated was two for limited disease, 3-4 for extensive disease with one metastatic site, 4 for two or three, and 5 for four or more sites of metastases. Based on a summation of pretreatment nucleoside/creatinine ratios, a discriminant for survival was derived giving curves separating patients (P = 0.086) similar to the discriminant based on stage of disease. Although discordant results were noted, an overall correlation of 75% agreement with clinical assessment was estimated in response categories when monitoring changes associated with therapy. PMID- 6291743 TI - Hepatitis B virus: pathogenesis and prevention of primary cancer of the liver. AB - The currently available data indicate that persistent infection of the liver with hepatitis B virus (HBV) is involved in the etiology and pathogenesis of nearly all primary hepatocellular carcinomas in humans. Immunization of high risk populations with the hepatitis B vaccine and prevention of mother-to-child transmission of HBV with hepatitis B immunoglobulin should prevent the development of chronic hepatitis B infection and, therefore, should also prevent the development of hepatocellular carcinoma. How HBV "causes" cancer of the liver is unknown. Research on the role of HBV in hepatocarcinogenesis may provide information on how HBV and other viruses induce cancer and may lead to the discovery of other virus-cancer relationships in humans. PMID- 6291744 TI - Adenoid cystic carcinoma involving the external auditory canal. A clinicopathologic study of 16 cases. AB - Sixteen patients with a rare tumor, adenoid cystic carcinoma (ACC) involving the external auditory canal, have been studied. Clinically, most patients complained of ear pain, often of several years duration. On physical examination, a mass or a nodule usually was identified in the ear canal. In most cases, treatment consisted of a wide surgical resection of the auditory canal and adjacent structures. Histologically, these neoplasms had the same appearance as ACC originating in salivary glands. In eight cases, the tumor was confined to the ear canal wall and unquestionably arose in this area, probably within the ceruminous glands. When incompletely excised, these lesions usually recurred locally. Nine patients had a total of 26 local recurrences. Most recurrences were found within two years of the preceding treatment, but the interval in some cases was long, ranging up to 14 years. Of the 16 patients, seven had no evidence of recurrence following surgical resection, two were living with recurrent, unresectable tumor, five had died of disease, one had died of other causes, and one was lost to follow-up. Some patients died of tumor after a prolonged clinical course with multiple recurrences. Death usually was caused by intracranial extension by the tumor, or by pulmonary metastases. Radiotherapy did not appear to cure the lesion, but probably resulted in palliation. An increased incidence of recurrent, unresectable tumor or of death from the neoplasm correlated with the following histologic features: demonstration of tumor on the lines of surgical excision, involvement of the parotid gland, extension into bone, perineural invasion and local recurrence of tumor. PMID- 6291745 TI - The clinical behavior of "mixed" small cell/large cell bronchogenic carcinoma compared to "pure" small cell subtypes. AB - Biopsy specimens from 19 previously untreated lung cancer patients were prospectively diagnosed as small cell carcinoma with a large cell component. The patients were thoroughly staged and received intensive combination chemotherapy. They represented 12% of all small cell carcinoma cases eligible for aggressive chemotherapy protocols during a 5.5 year period. To determine whether the clinical behavior of this "mixed" histologic variant differed from the other histologic subtypes of small cell lung cancer, we compared these 19 patients to a concurrent group of 103 patients with only small cell cancer in their diagnostic biopsies given equivalent therapy. The "mixed" histology patients were comparable to the "pure" small cell group in age, performance status, extent of disease, and frequency of bone marrow, liver, bone, and central nervous system metastases. Their complete plus partial response rare (58%) was significantly less than the response rate for the "pure" small cell patients (91%), their complete response rate was also lower (16 versus 46%), and their overall survival was significantly shorter (median, 6 versus 10.5 months) Mixed histology small cell/large cell carcinoma represents a distinct pathologic variant of small cell carcinoma of the lung, associated with lower response rates and shorter survival than the "pure" small cell subtypes. Since combination chemotherapy yields some complete responses and long-term disease-free survival in these patients, however, aggressive treatment with potentially curative intent should be considered in their management. PMID- 6291746 TI - Positive correlation between high aryl hydrocarbon hydroxylase activity and primary lung cancer as analyzed in cryopreserved lymphocytes. AB - Blood samples from closely monitored patients at the Veterans Administration Hospital in Houston, Texas, were collected, coded, and sent to Microbiological Associates over an 8-month period. Lymphocytes were isolated and cryopreserved at -190 degrees. Lymphocyte samples were simultaneously thawed, phytohemagglutinin activated, and analyzed for benz(a)anthracene-induced aryl hydrocarbon hydroxylase (AHH) levels, [3H]thymidine incorporation, and reduced nicotinamide adenine dinucleotide-dependent cytochrome b5 (cytochrome c) reductase activity. Determinations were made at both 96 and 120 hr in culture, and peak activities were compared among a total of 51 individuals who expressed such lesions as squamous cell carcinomas (22%), adenocarcinomas (14%), oat cell carcinomas (6%), chronic obstructive pulmonary disease (22%), and other nonmalignant diseases. Of the 14 highest AHH/cytochrome c activities observed, all were found in patients with primary lung cancer. Mean AHH/cytochrome c activities were 0.89 for lung cancer patients (a total of 21) and 0.47 for noncancer patients (a total of 30) (p less than 0.001). No relationship was observed between AHH/cytochrome c activity and age of patient, numbers of cigarettes smoked, family history of cancer, location or histological type of tumor, or level of phytohemagglutinin blastogenesis ([3H]thymidine cpm/cytochrome c). Whether the higher AHH levels are the cause or the result of the primary lung cancer remains to be determined. PMID- 6291747 TI - Induction of cytoskeleton-associated proteins during differentiation of human myeloid leukemic cell lines. AB - Alterations in the expression of proteins associated with the cytoskeletal framework during differentiation of two human myeloid leukemia cell lines were analyzed by two-dimensional gel electrophoresis of Triton-insoluble cellular framework fractions. During in vitro differentiation of HL60 (human promyelocytic leukemia line) and U937 (human monocytoid leukemia line), several new cytoskeleton-associated (CSK) proteins are induced. All of these CSK proteins are also present in freshly isolated normal granulocytes and macrophages. One of these differentiation-induced proteins comigrates with vimentin. There are several differentiation-sensitive proteins, i.e., those that are no longer synthesized upon differentiation. The changes in CSK protein synthesis during differentiation of HL60 and U937 cells do not seem to be related to drug treatment per se since exposure to conditioned medium from phytohemagglutinin stimulated lymphocytes as well as to dimethyl sulfoxide and 12-O tetradecanoylphorbol-13-acetate results in the production of many similar proteins. In vitro conditions that do not result in differentiation of HL60 and U937, such as cultivation in serum-free medium, do not induce the CSK proteins that we describe. A notable finding in this study is that all of the qualitative changes in the proteins synthesized during differentiation are detected in the cytoskeletal (Triton-insoluble) fraction, whereas only minor quantitative alterations are observed in the Triton-insoluble extract. The changes in CSK protein components occur in an orderly fashion. Vimentin, an intermediate filament protein, is synthesized in large amounts prior to changes in cellular morphology and the induction or loss of other CSK proteins. Vimentin may play an important role in the reorganization of the cytoskeleton to support the process of differentiation. The other CSK proteins are synthesized sequentially along with the morphological and functional changes during differentiation. These model systems, therefore, present an opportunity to investigate the role of specific cytoskeletal components in cellular differentiation. PMID- 6291748 TI - Anchorage-independent growth-conferring factor production by rat mammary tumor cells. AB - Conditioned medium from cultures of 7,12-dimethylbenz(a)anthracene-induced rat mammary tumor cells contain factors that resemble sarcoma growth factor and other transforming growth factors in biological activity but differ in their physical properties. The mammary tumor factors (MTF) are acid stable and heat and protease sensitive. They inhibit the binding of epidermal growth factor, but not insulin, to mouse embryonal carcinoma cells. MTF confers upon normal rat kidney and BALB/c 3T3 cells the ability to grow in soft agar. This effect is enhanced synergistically by high concentrations of fetal calf serum but not by epidermal growth factor. Anchorage-independent growth promotion, however, is not seen with normal mammary epithelial cells, although MTF is mitogenic for these cells as well as normal rat kidney cells, BALB/c-3T3 cells, and chick embryo fibroblasts in monolayer culture, MTF is not mitogenic for primary cultures of the tumor cells from which the factors are derived. Two major molecular weight species of MTF, eluting at Mr 6,000 and 65,000 to 70,000 on Bio-Gel P-100 columns, are present in acid-ethanol extracts of 7,12-dimethylbenz(a)anthracene- and nitrosomethylurea-induced rat mammary tumors. Transplantable tumors derived from primary 7,12-dimethylbenz(a)anthracene- or nitrosomethylurea-induced tumors have little or no MTF activity. These results demonstrate that different chemically induced rat mammary tumors contain transforming growth factor-like activities. Furthermore, it is possible that MTF is unnecessary for the maintenance of tumorigenicity, since some tumors contain no detectable MTF. PMID- 6291749 TI - Neoplastic transformation and defective control of cell proliferation and differentiation. AB - The control of proliferation of nontransformed 3T3 t-proadipocytes in vitro can be mediated at three states in the G1 phase of the cell cycle. These states are induced by the commitment of cells to differentiate (GD); by growth factor deprivation at low density or "contact inhibition" at high density (Gs); and by nutrient deprivation (GN). To determine if neoplastic transformation of proadipocytes is associated with a selective defect in one or more of these G1 growth arrest processes, we developed and studied eight cloned and several noncloned tumorigenic proadipocyte cell lines. We report that all transformed proadipocyte cell lines are tumorigenic and all lack the ability to arrest at GD and differentiate. By contrast, or approximately 90% of transformed proadipocyte cell lines retain their ability to growth arrest at Gs at low density when deprived of growth factors, and or approximately 90% growth arrest at GN when deprived of nutrients. These observations suggest that neoplastic transformation of proadipocytes is primarily associated with abrogation of growth control mediated at GD. However, whereas most transformed proadipocytes arrest at Gs at low density when deprived of serum, all transformed proadipocyte cell lines do not efficiently arrest at Gs at high density due to "contact inhibition." This suggests that neoplastic transformation of proadipocytes results from a primary defect in growth control mediated at GD and from an additional defect at Gs. These results are discussed with respect to their possible significance for the biological mechanisms of the initiation and promotion of carcinogenesis. PMID- 6291750 TI - Hepatitis B virus and cigarette smoking: risk factors for hepatocellular carcinoma in Hong Kong. AB - One hundred seven Chinese patients with primary hepatocellular carcinoma (PHC) were compared with 107 hospital controls for the presence of hepatitis B surface antigen and smoking, drinking, and dietary habits. Eighty-two % of PHC cases were hepatitis B surface antigen positive compared to 18% of controls (relative risk, 21.3; 95% confidence limits, 10.1 and 45.9). Prior history of jaundice was significantly related to PHC, independent of hepatitis B surface antigen status. There was a significant association between cigarette smoking and PHC negative for hepatitis B surface antigen. The relative risk of hepatitis B surface antigen negative PHC for heavy smokers (20 + cigarettes/day) was 3.3 compared to light smokers and nonsmokers (95% confidence limits, 1.0 and 13.4). Our data indicated that infection by the hepatitis B virus and cigarette smoking were independent risk factors for PHC. PMID- 6291751 TI - Clonogenic assay for Wilms' tumor: improved technique for obtaining single-cell suspensions and evidence for tumor cell specificity. AB - Wilms' tumors from seven patients were dissociated by mechanical and enzymatic means; this technique resulted in single-cell suspensions for five specimens and a few aggregates for two. By dye exclusion, cell viability ranged from 56 to 100% (median, 92%). All seven preparations produced more than five colonies/2 x 10(5) cells plated. Forty-three colonies grown from cells of a glucose-6-phosphate dehydrogenase heterozygote were of the same glucose-6-phosphate dehydrogenase isoenzyme type as the original tumor, indicating that the assay is specific for tumor cells. We attribute the high rate of colony formation to an improved method of cell preparation (combined mechanical and enzymatic dissociation of tumors) which may be applicable to other primary human tumors assayed in the soft agar system. PMID- 6291752 TI - Benzo(a)pyrene and 7,12-dimethylbenz(a)anthracene metabolism and DNA adduct formation in primary cultures of hamster epidermal cells. AB - Primary cultures of hamster epidermal cells exposed to hydrocarbon, 1 microgram/ml, rapidly metabolized [3H]benzo(a)pyrene and [14C]7,12 dimethylbenz(a)anthracene to ethyl acetate:acetone- and water-soluble metabolites. By 24 hr, only 13.6% of the organic solvent-soluble radioactivity recovered in the medium was unchanged [3H]benzo(a)pyrene, and only 5.9% was unchanged [14C]7,12-dimethylbenz(a)anthracene. With both hydrocarbons, the major water-soluble metabolites found extracellularly were conjugated with glucuronic acid; these were primarily phenolic derivatives. Metabolites cochromatographing with 7,8-dihydro-7,8-dihydroxybenzo(a)pyrene or trans-3,4-dihydro-3,4-dihydroxy 7,12-dimethylbenz(a)anthracene were not detectable in high-pressure liquid chromatographic profiles of organic solvent-soluble intracellular and extracellular metabolites. However, analysis of [3H]benzo(a)pyrene: and [3H]7,12 dimethylbenz(a)anthracene: DNA adducts indicated that these putative proximate carcinogenic metabolites were formed in these cells and subsequently metabolized to DNA-binding products. The results suggest that metabolic incompetence may not be an explanation for the relative resistance of the hamster to epidermal carcinogenesis by polycyclic hydrocarbons. PMID- 6291753 TI - Promotion of liver cancer development by brief exposure to dietary 2 acetylaminofluorene plus partial hepatectomy or carbon tetrachloride. AB - Adult male Fischer rats were exposed to a necrogenic dose (200 mg/kg) of diethylnitrosamine or to nonnecrogenic doses of N-methyl-N-nitrosourea, 1,2 dimethylhydrazine, or benzo(a)pyrene following partial hepatectomy or sham hepatectomy. This treatment by itself led to no hepatocellular carcinomas by 8 to 18 months, except in animals given N-methyl-N-nitrosourea, which showed a 30% incidence by 12 months. With each treatment regimen, exposure to dietary 2 acetylaminofluorene for 2 weeks coupled with partial hepatectomy or the administration of a necrogenic dose of CCl4, was associated with an incidence of 68 to 94% of cancer at 8, 12, or 18 months, depending upon the initiating carcinogen used. Appropriate controls showed either no hepatocellular carcinoma or a much lower incidence. It is concluded that the 2-week exposure to dietary 2 acetylaminofluorene plus partial hepatectomy or the administration of CCl4 has a strong promoting effect on liver carcinogenesis with four different chemical carcinogens. PMID- 6291754 TI - Antibody reacting with the murine mammary tumor virus in the serum of patients with breast carcinoma: a possible serological detection method for breast carcinoma. AB - Sera from patients with Stages A and B infiltrating ductal carcinoma of the breast, benign breast disease, cancers other than breast carcinoma, and normal female controls were examined by indirect immunoelectron microscopy (IEM) and a viral agglutination test for evidence of antibodies directed against murine mammary tumor virus (MMTV). Sera from 41 (79%) of 52 patients with breast carcinoma and eight (19%) of 42 normal subjects or patients with benign breast disease (noncancer subjects) showed evidence of MMTV labeling by IEM. In the MMTV agglutination test, significant virus agglutination (2+ to 4+) was present in eight (13%) of 61 noncancer sera, 58 (86%) of 68 breast carcinoma sera, and two (11%) of 18 other cancer sera. The results of the more rapid MMTV agglutination test correlated well with IEM. Analysis of reacting antibody by IEM revealed no immunoglobulin A and significant immunoglobulin M and immunoglobulin G antibody. Serum reactivity against MMTV was completely absorbed by MMTV but not by the glycoprotein with a molecular weight of 52,000 of MMTV, Friend murine leukemia virus, avian myeloblastosis virus, or sheep erythrocytes. It is concluded that reactivity of human antibodies to MMTV is strongly associated with, but is not entirely specific for, breast carcinoma. It remains to be determined if normal persons with these antibodies will ultimately develop breast cancer and should therefore be considered at high risk. These tests may have potential usefulness as a diagnostic screen for breast cancer. PMID- 6291755 TI - Compartmentalization of regulatory subunits of cyclic adenosine 3':5' monophosphate-dependent protein kinases in MCF-7 human breast cancer cells. PMID- 6291756 TI - Induction of murine tumors in adult mice by a combination of either avian sarcoma virus or human adenovirus and syngeneic mouse embryo cells. AB - Primary murine Rous sarcoma was produced in adult mice of seven strains, C57BL/6, DBA/2, BALB/c, C3H/He, CBAJ, AKR, and DDD, by s.c. inoculation of a mixture of 5 X 10(6) chicken tumor cells containing Schmidt-Ruppin Rous sarcoma virus and 9- to 12-day-old mouse embryo cells (MEC) (2 X 10(6) ) of the syngeneic strain. The sarcoma developed at the site of injection in almost all mice tested, but there were some differences in the latent period and the survival time among mouse strains. When the number of cells inoculated was reduced to 5 X 10(4) for chicken tumor cells induced by the Schmidt-Ruppin strain of Rous sarcoma virus (SR-CTC) and 2 X 10(4) for MEC, no tumor was produced in C3H/He mice. These tumors had strain specificity and the Schmidt-Ruppin strain of Rous sarcoma virus genome in masked form. The tumor at the site of injection originated in the embryo cells injected along with SR-CTC. This was confirmed by CBAT6/T6 marker chromosome analysis of the tumor cells of CBA mice induced with SR-CTC plus CBAT6/T6 MEC and also confirmed by transplantation of a C57BL/6 X C3H/He F1 tumor which had been induced with SR-CTC plus C3H/He or C57BL/6 MEC. Tumor induction in adult mouse by a mixture of virus and syngeneic 9- to 14-day-old embryo cells was tested for human adenovirus serotype 12 (Ad12) and simian virus 40. Primary Ad12 tumor was also induced in adult CBA, C3H/He, and DDD mice by 4 X 10(5 to 6) 50% tissue culture infective dose of Ad12 with 5 X 10(6) syngeneic embryo cells. This tumor contained Ad12 T-antigen-positive particles in cells. But in the case of simian virus 40, the tumor did not appear for about 300 days of observation. PMID- 6291757 TI - Characterization of the microsomal cytochrome P-450 species induced in rat liver by 2-acetylaminofluorene. AB - 2-Acetylaminofluorene induces the level of cytochrome P-450 in rat liver microsomes by 50% (p less than 0.001). This induced cytochrome(s) was characterized and compared to the major forms of cytochrome P-450 induced by phenobarbital and 3-methylcholanthrene. The properties investigated were: the absorption maximum of the complex formed between reduced cytochrome P-450 and carbon monoxide; the substrate specificities using aminopyrine, benzphetamine, ethylmorphine, benzo(a)pyrene, ethoxycoumarin, ethoxyresorufin, and 2 acetylaminofluorene itself as substrates; metabolite patterns with benzo(a)pyrene and 2-acetylaminofluorene; sensitivity to different inhibitors; binding spectra with aniline and hexobarbital; and molecular weight as determined by sodium dodecyl sulfate:disc gel electrophoresis. The results indicate that 2 acetylaminofluorene induces a form(s) of cytochrome P-450 especially effective in the metabolism of this substance itself (i.e., the process can be called substrate induction) and different from the major forms of cytochrome P-450 induced by phenobarbital and 3-methylcholanthrene. PMID- 6291758 TI - Fatty acid-dependent benzo(a)pyrene oxidation in colonic mucosal microsomes: evidence for a distinct metabolic pathway. PMID- 6291759 TI - Characterization of a cyclic nucleotide-independent protein kinase highly active in human adrenocortical carcinoma. AB - Study of the protein kinase activity pattern of four human adrenocortical carcinoma showed that in all the samples examined a histone kinase (HK III) activity was present at high level, whereas it was barely detectable in normal tissue. HK III was separated from other known adrenocortical protein kinases by diethylaminoethyl cellulose chromatography. Isolated HK III exhibited a histone (H2B) protamine-phosphotranferase selectivity and used adenosine triphosphate but not guanosine triphosphate as phosphate donor. Serine was identified as the only target amino acid phosphorylated in the protein substrate. HK III showed an apparent molecular weight of 65,000 upon gel filtration and an apparent sedimentation coefficient of 3.7S. HK III activity was cyclic adenosine 3':5' monophosphate independent and was not influenced by calcium, calmodulin, polyamines, and heparin. The significance of HK III activity in adrenocortical carcinoma extracts at a high level as compared to that of normal tissue remains to be clarified with regard both to its possible relationship with tumoral cell growth and differentiation processes and to its potential interest as a marker of human tumoral tissue activity. PMID- 6291760 TI - Detection and biochemical characterization of antigens in human leukemic sera that cross-react with primate C-type viral proteins (Mr 30,000). AB - Antigens have been detected in 35 to 40% of sera from patients with leukemia that cross-react with the Mr 30,000 core proteins (p30) of baboon endogenous virus (BaEV) and/or of simian sarcoma-simian sarcoma-associated virus (SiSV) in a solid phase enzyme immunoassay using anti-SiSV p30 and anti-BaEV p30 antisera. These antigens could not be found in sera from nonleukemic persons. Fetal calf serum; normal chicken, goat, rabbit, and rhesus sera; and normal human serum components like albumin, immunoglobulins, and transferrin did not react with the anti-SiSV p30 and anti-BaEV p30 antisera. The reactivity in the leukemic sera was abolished by treatment with protease, but not with glycosidases. The antigens purified by immunoaffinity chromatography showed essentially one band with an apparent molecular weight of 70,000 on sodium dodecyl sulfate: polyacrylamide gel electrophoresis. In competition enzyme-linked immunosorbent assays the leukemia associated antigens competed out SiSV p30 in the anti-SiSV p30 system. Peptide mapping experiments with antigens from sera of two different leukemic patients showed that the two antigens were identical concerning numbers of peptides and their position. Eleven of 21 major peptides of SiSV p30 and 10 of 20 major peptides of BaEV p30 (50 to 60% of major peptides) showed mobilities identical with those of the major peptides of the human antigens. The data suggest the presence in human sera of retroviral antigens closely associated with leukemia. PMID- 6291761 TI - Comparative studies on the quantitative analysis of experimental metastatic capacity. AB - The purpose of these studies was to establish a procedure for determining the relative experimental metastatic potential of unrelated murine tumors. We used three tumors (the B16-F10 melanoma, which is syngeneic to the C57BL/6N mouse, and the K-1735 melanoma and the UV-2237 fibrosarcoma, which are syngeneic to the C3H/HeN mouse). Various numbers of tumor cells were injected into normal or immunosuppressed syngeneic recipients and into 3-week-old BALB/c nude mice. At appropriate intervals, the recipient mice were killed, and the metastatic burden was determined. The number of experimental metastases was not linearly correlated with cell input. Thus, simply comparing the incidence of metastasis resulting from the injection of one predetermined dose of tumor cells did not allow for determination of their relative metastatic capacities. More reproducible and meaningful results were obtained by introducing increasing numbers of viable tumor cells admixed with a constant number of nontumorigenic (X-irradiated) tumor cells serving as carrier. The incidence of metastasis by few or many injected cells is influenced by host factors such as immune status, and therefore determinations of the true metastatic nature of any given tumor necessitate the choice of an appropriate recipient. PMID- 6291762 TI - The physiology and pharmacology of urinary tract dysfunction. PMID- 6291763 TI - Random prospective study cyclophosphamide, doxorubicin, and methotrexate (CAM) combination chemotherapy versus single-agent sequential chemotherapy in non small cell lung cancer. AB - A group of 104 patients with unresectable non-small cell lung cancer were randomized to receive combination chemotherapy with cyclophosphamide, doxorubicin, and methotrexate (CAM) or single-agent sequential chemotherapy with the same three agents. CAM combination chemotherapy produced a 22% objective response rate, including two complete remissions, compared to a 9% response rate, including one complete remission, produced by single-agent therapy (P = 0.16). The median survival time was 32 weeks (range, 3-116) for CAM, compared to 25 weeks (range, 4-179 +) for sequential single agents (P = 0.24). Overall survival was 31% (1-year), (16%) (1 1/2-year), and 5% (2-year), with no difference between the study arms. Although there was no statistically significant survival advantage for the CAM arm, both arms had survival superior to that in historical controls, presumably because of better patient selection. This study indicates that cyclophosphamide, doxorubicin, and methotrexate are, at best, marginally active as single agents, and new drugs with more efficacy are needed before combination chemotherapy can be expected to result in any meaningful prolongation of survival in non-small cell lung cancer. PMID- 6291764 TI - Fine structure and function of isolated gonadotropic cells as revealed from pituitaries of immature rainbow trout, Salmo gairdneri, by means of a new enzymatic dispersion technique. AB - A procedure has been developed for dissociating pituitary glands of juvenile rainbow trout, Salmo gairdneri, producing a preparation of single dispersed pituitary cells in which morphological and functional integrity is preserved. The pituitaries are dispersed by sequential treatment with 0.1% collagenase, 0.04% ethylene-diamine-tetra-acetic acid (EDTA) and 0.125% dispase. The cell yield is 0.3--0.35 x 10(6) cells per pituitary with a cell viability percentage of 95 +/- 1% and single cell percentage of 87 +/- 4%. The isolated cells are kept in a suspension system and the gonadotropic cells are identified by the double antibody immuno-enzyme-cytochemical technique using anti-carp-beta-gonadotropin as first antibody. Secretory activity is estimated by measuring the gonadotropin content in cells and culture media by radioimmunoassay. Isolated cells show an autonomy of gonadotropin secretion. 17 alpha-Methyltestosterone both in vivo and in vitro stimulates the production of gonadotropin in the cells and seems to inhibit its release from the cells. It is concluded that this in vitro system can be used as a model for studying the control of gonadotropic cells in juvenile rainbow trout. PMID- 6291765 TI - Alpha- and beta-receptor control of catecholamine secretion from isolated adrenal medulla cells. AB - The regulation of secretion of catecholamines from bovine adrenal medulla cells was investigated by use of an improved and highly efficient method for isolating viable and responsive cells from this tissue. The method involves an in situ collagenase perfusion affecting only the connective tissue matrix of the medulla while leaving the cortex intact. The cells released both epinephrine and norepinephrine in response to stimulation by 100 microM acetylcholine. The ratio of epinephrine to norepinephrine in the medium following non-stimulated (basal) release, was similar to that found in the intact cells. On the other hand, a lower ratio of epinephrine to norepinephrine was found in the medium following stimulation by acetylcholine due mainly to preferential secretion of norepinephrine. This release ceased after 15 min of incubation and consisted of 15--20% of the catecholamines initially present in the cells. Exogenous epinephrine was found to inhibit total catecholamine secretion; however, it stimulated norepinephrine release. Addition of isoproterenol caused a stimulation of release while propranolol was inhibitory. Norepinephrine inhibited total release not favoring any specific catechol. Other alpha-agonists, such as clonidine, also had an inhibitory effect. These results suggest a receptor mediated mechanism for the fine regulation of secretion from the adrenal medulla. PMID- 6291766 TI - Ca2+ transporting activity of membrane fractions isolated from the post mitochondrial supernatant of rat liver. AB - The post-mitochondrial supernatant of rat liver contains two vesicular fractions which transport Ca2+ actively. The heavier fraction, sedimenting at 17.500 xg, 20 min, is enriched in plasma membrane markers and apparently contains both a Ca2+ pumping ATPase and a Na+/Ca2+ exchanger. These activities have been attributed to the plasma membrane vesicles. The lighter fraction, sedimenting at 100.000 xg, 60 min, is enriched in endoplasmic reticulum markers, and contains only a Ca2+ pumping ATPase, which can be differentiated from that of the heavier fraction on the basis of the sensitivity to vanadate. The Ca2+ pumping activity of endoplasmic reticulum appears to be regulated by both a cAMP-dependent, and a calmodulin-dependent system. The former system involves a heat-stable protein fraction from the cytosol. The regulation by the cAMP and the calmodulin dependent systems involves the phosphorylation of several proteins in the endoplasmic reticulum membrane. PMID- 6291767 TI - Regulation of the synthesis of extracellular matrix components in chondroblasts transformed by a temperature-sensitive mutant of Rous sarcoma virus. AB - Regulation of cartilage extracellular matrix synthesis has been examined in chondroblasts infected with a temperature-sensitive mutant of Rous sarcoma virus. Cells grown at the nonpermissive temperature synthesized large amounts of several chondroblast-specific polypeptides (type IV proteoglycan core protein, type II procollagen, a proteoglycan link protein(s) and a 60 kd protein) and very low levels of fibronectin. At the permissive temperature, synthesis of chondroblast specific proteins was coordinately reduced, while fibronectin synthesis was greatly increased. These changes reflected comparable alterations in levels of translatable mRNAs encoding these proteins. This analysis also revealed the unexpected presence in the transformed cells of type I collagen mRNAs, which are not used in intact cells, indicating that a posttranscriptional control (or controls) may be acting in transformed chondroblasts. PMID- 6291768 TI - Epstein-Barr virus superinduces a new human B cell differentiation antigen (B LAST 1) expressed on transformed lymphoblasts. AB - We have developed a monoclonal antibody that detects the first human B-cell specific differentiation antigen expressed on transformed B lymphoblasts. The antigen is termed B-LAST 1 and is found on B cells transformed in vitro with Epstein-Barr virus (EBV) and pokeweed mitogen; in vivo with EBV and antigen; and on neoplastic B cells from chronic lymphocytic leukemia and poorly differentiated lymphoma. The antigen has a molecular weight of 45,000 and was not found on cells of T, null or myeloid lineage, whether obtained from peripheral blood, lymph nodes, neoplasms or cell lines. The antigen is expressed at a much higher level on EBV-infected cells than on any other cell type studied, but does not appear to be virally encoded. The significance of this antigen in the process of viral and nonviral transformation and its possible role as a target for T-cell-mediated immunity against virus-infected cells is discussed. PMID- 6291769 TI - Initiation of replication at specific origins in DNA molecules microinjected into unfertilized eggs of the frog Xenopus laevis. AB - Initiation of DNA replication at specific origins was observed by electron microscopy after microinjection of pXlr11, pXlr14 or Col E1 plasmid DNA molecules into unfertilized eggs of the frog, Xenopus laevis. These results are in apparent contradiction with published reports (Harland and Laskey, Cell 21, 761-771, 1980; Laskey and Harland, Cell 24, 283-284, 1981) that specific origin sites were not used in Xenopus laevis eggs. We suggest that eucaryotic origins exist that both increase the probability of replication of contiguous sequences and determine the site at which replication is most likely to begin. PMID- 6291770 TI - Role of small t antigen in the acute transforming activity of SV40. AB - A plasmid, pHR402, containing SV40 sequences that include a truncated early region bearing an intact t-coding sequence and a functionally intact late region, was introduced into thymidine kinase deficient (tk-) mouse L cells by cotransformation with a cloned tk gene. tk+ cotransformants synthesized SV40 t but not T antigen, and no truncated T-coding sequence products were detected. The viral sequences of pHR402 were reconstituted as a virus in COS1 cells, and acute infection of untransformed mouse cells with this viral stock (SV402) also led to the appearance of t but not T or a truncated T. Abortive transformation assays of such infected cells were negative, as were those performed on the same cells infected with either of two viral mutants (dl883 and dl884), each of which leads to T but not t synthesis. However, mixed infection with SV402 and either dl883 or dl884 led to a clear abortive and permanent transformation response. Thus, at least in part, t and T appear to function in a complementary fashion in eliciting transformation expression by SV40-infected cells. PMID- 6291771 TI - SV40 large tumor antigen can regulate some cellular transcripts in a positive fashion. AB - Eleven cDNA clones identified from a cDNA library prepared from the mRNA fraction of SV40 transformed cells detected, by hybridization, higher levels of cellular mRNA in SV40-transformed cells than in nontransformed cells. Three of these cDNA clones detected levels of cellular mRNA that were more than 100-fold greater in SV40tsA transformed cell lines grown at the permissive temperature than in those grown at the nonpermissive temperature. Northern blot hybridizations confirmed these results and in some cases detected RNA species of multiple sizes that were regulated in a temperature-dependent fashion in SV40tsA transformed cell lines. Infection of 3T3 cells with SV40 stimulated the levels of RNAs complementary to these cDNA clones. The results demonstrate that the SV40 large T antigen can regulate the steady state levels of some cellular RNA species. PMID- 6291772 TI - Persistence of freely replicating SV40 recombinant molecules carrying a selectable marker in permissive simian cells. AB - We have demonstrated that a SV40-pBR322 recombinant vector (pSV2-gpt) carrying a bacterial gene of selectable phenotype (Eco-gpt) may persist extrachromosomally in COS1 cells, a simian cell line that endogenously produces SV40 large T antigen. The amount of circular (supercoiled) recombinant DNA was estimated to be between 5 and 2000 copies per cell among several pSV2-transformed COS1 clonal lines examined. Complete pSV2 molecules were found in the majority of the transformants, although some of the pSV2 DNAs recovered were shown to have deletions in the pBR322 region. Our results indicate that removal of the pBR322 "inhibitory sequence" in pSV2 is not necessary for stable maintenance of these recombinant molecules in COS1 cells. In addition, large amounts of pSV2-related high molecular weight DNAs, probably concatemers of pSV2, were detected in the transformed lines. PMID- 6291773 TI - Regulation of white locus expression: the structure of mutant alleles at the white locus of Drosophila melanogaster. AB - We have analyzed the structures of 19 mutant alleles at the white locus of Drosophila melanogaster. Thirteen of the mutant alleles in our selected sample arose spontaneously, and of these, seven are associated with insertions of non white-region DNA sequence elements. Several lines of evidence strongly suggest that these insertions are responsible for their associated mutant alleles, and further suggest that most or all of these insertions are transposons. Moreover, the white locus DNA sequences can be divided into two nonoverlapping domains on the basis of the properties of the two domains as mutational targets. One of these domains behaves, in this regard, in the manner expected of functional coding sequences, whereas the other does not. We propose a model for the nature and function of the presumptive noncoding white locus genetic elements. The two domains of the white locus defined by our studies are approximately coextensive with the functionally distinct subintervals of the locus defined by previous genetic analysis. Lastly, our results strongly suggest that the dominant, mutable wDZL allele results from the insertion of a transposon outside of, but near, the white locus. This putative transposon apparently carries genetic elements that act at a distance to repress expression of the white locus. PMID- 6291774 TI - The unstable wDZL mutation of Drosophila is caused by a 13 kilobase insertion that is imprecisely excised in phenotypic revertants. AB - We have analyzed the lesion in wDZL, a genetically unstable mutant allele of the eye color locus, white, of Drosophila melanogaster. We have cloned the DNA of the white locus region of flies carrying the wDZL allele and find a 13 kilobase insertion not present in the wild-type at the corresponding location. In 12 independent cases examined, reversion to a wild-type eye color phenotype correlates with the excision of a portion of this 13 kilobase insertion, indicating that the insertion is the cause of the mutation. The portion of the insertion that is excised in these eye color revertants is heterogeneous in size but appears to include the central 6 kilobases of the insertion in all cases. Many of these eye color revertants continue to undergo mutation at the white locus, indicating that the residual portion of the insertion in these revertants is sufficient to promote mutations. PMID- 6291775 TI - FB elements are the common basis for the instability of the wDZL and wC Drosophila mutations. AB - The DNA insertions that cause the highly unstable mutations wC and wDZL share extensive homology with the FB family of transposable elements. FB elements carry long, internally repetitious, inverted terminal repeats and thus differ in structure from other transposable elements. Our results suggest that FB elements may excise and cause chromosomal rearrangements at unusually high frequencies. The wC insertion is a single FB element. The wDZL insertion differs in that it contains two FB elements, one at each terminus. The wC and wDZL insertions contain 4.0 and 6.5 kilobase nonhomologous segments between their terminal repeats. In contrast to the middle repetitive FB elements, the central segment of the wDZL insertion is single-copy and present at a fixed location in the wild type genome. It has apparently been transposed by the action of flanking FB elements, causing the wDZL mutation at its new location. PMID- 6291776 TI - A hierarchic arrangement of the repetitive sequences in the Balbiani ring 2 gene of Chironomus tentans. AB - One cloned cDNA sequence, pCt63, was used to characterize the repeated structure of the Balbiani ring 2 gene in Chironomus tentans. Although small in size (0.63 kb), the cDNA insert corresponds to a large portion (25 kb) of the BR2 gene (37 kb). Southern blotting experiments suggested that a large part of the BR2 gene consists of tandemly repeated units, each about 215 bp. Sequence analysis of the cDNA confirmed the repeated nature of the BR2 gene and revealed the internal structure of the repeat unit. Each such unit is composed of two regions of approximately equal length; one is highly ordered and built from about six 18 bp repeats, each consisting of a slightly diverged 9 bp duplication. The recorded hierarchic arrangement of the repetitive sequences in the BR2 gene and a specific pattern of base substitutions along the gene have enabled us to propose how a major part of the giant BR2 gene has evolved from a short primordial sequence, 110-120 bp in length. PMID- 6291777 TI - T-DNA organization in homogeneous and heterogeneous octopine-type crown gall tissues of Nicotiana tabacum. AB - Octopine-type tumor tissue was obtained both by infection of plants or isolated protoplasts with Agrobacterium tumefaciens and by somatic hybridization of normal and crown gall tobacco cells. Analysis of T-DNA by Southern blotting of clones and uncloned tissue reveals that, whereas tumors induced on plants are heterogeneous mixtures of cells differing in T-DNA organization, each tissue derived from transformed protoplasts or from somatic hybridization is homogeneous. Detailed analysis of T-DNA organization showed that TL- or "core" T DNA was always present at one or two copies per diploid genome. However, sometimes it was present in a modified form, either deleted, extended, tandemly duplicated or probably methylated. TR-DNA was not detected. The observed variation in the organization of T-DNA in octopine crown gall tissue did not appear to be a characteristic of the way the tissue was derived. PMID- 6291778 TI - Specific protein-DNA interaction at four sites flanking the chicken lysozyme gene. AB - Cloned DNA containing 22.2 kb of the chicken lysozyme gene region was screened with use of a nitrocellulose filter binding technique for specific recognition by nuclear DNA binding proteins from chicken oviduct cells. The analysis showed specific retention of four restriction fragments (BS1-BS4), which map approximately 6.1 and 3.9 kb upstream from the transcription start, and 2.8 and 6.2 kb downstream from the poly(A)-addition point of the lysozyme gene. The four DNA fragments mutually served as efficient competitors, indicating that only one class of proteins is involved in the recognition of all four sites. An apparent binding constant of KD = 6 X 10(-12) M was estimated for one of the binding fragments (BS1). Fine mapping of this fragment resolved two closely spaced contact areas at least 43 bp apart. PMID- 6291779 TI - Structure and properties of the bacterial nucleoid. PMID- 6291780 TI - Sodium channels in electrically excitable cells. PMID- 6291781 TI - Posttranslational processing of the LDL receptor and its genetic disruption in familial hypercholesterolemia. AB - Synthesis of the low density lipoprotein (LDL) receptor was studied by incubation of cultured human fibroblasts with 35S-methionine followed by immunoprecipitation with a monoclonal antireceptor antibody. The receptor was synthesized as a precursor with an apparent molecular weight of 120 kilodaltons (kd) that was converted to a mature form of 160 kd. This novel form of processing occurred 15 30 min after synthesis and did not appear to be due to the simple addition of N linked oligosaccharide chains. Fibroblasts from a child with the phenotype of homozygous familial hypercholesterolemia showed a disruption in receptor processing. This child has two different mutant alleles at the LDL receptor locus. One allele, inherited from his heterozygous mother, produces an abnormal 120 kd protein that cannot be processed to the mature 160 kd form. The other allele, inherited from his heterozygous father, produces a receptor that is synthesized as an elongated 170 kd precursor which undergoes a 40 kd increase in molecular weight to form an abnormally large receptor of 210 kd. PMID- 6291782 TI - Anti-VPg antibody inhibition of the poliovirus replicase reaction and production of covalent complexes of VPg-related proteins and RNA. AB - Anti-VPg antibodies inhibited host-factor-dependent RNA synthesis by the poliovirus replicase but not oligo(U)-primed synthesis, implicating VPg in the de novo initiation of replicase products. Complexes of VPg-related polypeptide and newly made RNA could be immunoprecipitated by anti-VPg antibody from the host factor-stimulated products of the replicase reaction. The complexes appeared to be covalently linked and involved 50 to 150 nucleotide chains of RNA that were RNAase-T1-resistant and could be largely poly(U). PMID- 6291783 TI - Expression from cloned cDNA of cell-surface secreted forms of the glycoprotein of vesicular stomatitis virus in eucaryotic cells. AB - A cDNA clone of the mRNA encoding the glycoprotein (G) of vesicular stomatitis virus was inserted into plasmid vectors under the control of either the SV40 early promoter (pSV2G) or the SV40 late promoter (pSVGL). Synthesis of G protein was observed in mouse L cells injected with pSV2G DNA or in COS1 cells transfected with pSVGL DNA. Immunofluorescent staining of G protein produced in both cell types showed a pattern of internal and cell-surface staining indistinguishable from that seen in cells infected with vesicular stomatitis virus. The G protein produced in transfected COS1 cells was the size of normal G protein and was glycosylated. Expression of a G protein lacking 79 amino acids from the COOH terminus was also examined. This G protein lacks the transmembrane domain and the hydrophilic COOH terminus, which, we postulated, anchor G protein in the lipid bilayer. This "anchorless" protein is glycosylated and is secreted, albeit slowly. PMID- 6291784 TI - Nucleotide sequence of Fujinami sarcoma virus: evolutionary relationship of its transforming gene with transforming genes of other sarcoma viruses. AB - We determined the entire nucleotide sequence of the molecularly cloned DNA of Fujinami sarcoma virus (FSV). The sequence of 1182 amino acids was deduced for the FSV transforming protein P130, the product of the FSV gag-fps fused gene. The P130 sequence was highly homologous to the amino acid sequence obtained for the gag-fes protein of feline sarcoma virus, supporting the view that fps and fes were derived from a cognate cellular gene in avian and mammalian species. In addition, FSV P130 and p60src of Rous sarcoma virus were 40% homologous in the region of the carboxyterminal 280 amino acids, which includes the phosphoacceptor tyrosine residue. These results strongly suggest that the 3' region of fps/fes and src originated from a common progenitor sequence. A portion (the U3 region) of the long terminal repeat of FSV DNA appears to be unusual among avian retroviruses in its close similarity in sequence and overall organization to the same region of the endogenous viral ev1 DNA. PMID- 6291785 TI - Continuing kappa-gene rearrangement in a cell line transformed by Abelson murine leukemia virus. AB - A cell line transformed by Abelson murine leukemia virus, called PD, is capable of carrying out kappa-gene rearrangement while growing in culture. Subclones of PD have diverse kappa-gene structures, and some derivatives show evidence of continued joining activity after as many as three subclonings. Analysis of PD sublineages has shown that a rearranged chromosome can undergo secondary kappa gene rearrangements, producing either a new rearrangement or a deletion of C kappa. Although the PD line actively rearranges its kappa genes, its rearranged heavy-chain genes show little variation, and there is no rearrangement of lambda genes. In PD subclones, DNA fragments representing the reciprocal product of kappa-gene rearrangement are often evident, and they may undergo either further rearrangement or deletion. The implications of multiple rearrangements on a single chromosome and of the maintenance of reciprocal fragments are considered in the context of a model that postulates that the V kappa and J kappa segments are not all organized in the DNA in the same transcriptional direction, leading to inversions rather than deletions during joining. PMID- 6291786 TI - Control of Tn5 transposition in Escherichia coli is mediated by protein from the right repeat. AB - The right repeat in Tn5, which encodes protein absolutely required for transposition, is also capable of inhibiting Tn5 transposition. Analysis of Tn5 mutants indicates that the left repeat is defective in supplying the transposition-inhibition function because of the sequence difference between the repeats located at nucleotide 1443; that the transposition-inhibition activity is a function of the quantity of right-repeat protein synthesis; that the smaller of the right-repeat proteins, protein 2, is sufficient for supplying the transposition-inhibition function (but not for the transposase activity); and that the transposition-inhibition function can act in trans, as opposed to the transposase activity, which functions efficiently only in cis. Gene fusion experiments indicate that the transposition-inhibition activity cannot be explained by autogenous regulation of right-repeat protein synthesis. Finally, immunoprecipitation assays of right-repeat protein-lacZ fusion proteins indicate that protein 2 is synthesized in significantly greater amounts than protein 1 in whole cells. This synthetic ratio may ber important with respect to the control of Tn5 transposition. PMID- 6291787 TI - Regulation of Tn5 by the right-repeat proteins: control at the level of the transposition reaction? AB - The transposon Tn5 consists of inverted repeats, called IS50R and IS50L, each of which encode two proteins. We show here that the larger protein encoded on IS50R, protein 1, is absolutely required for transposition. Deletion or insertion mutants that fail to make this protein fail to promote gene movement. In addition, his protein acts in cis preferentially. We also show that the smaller protein encoded on IS50R, protein 2, is competent to inhibit transposition of a Tn5 freshly introduced into the cell on a lambda phage. In contrast, the proteins from IS50L possess neither of these two activities. By assaying expression of proteins that are hybrids between beta-galactosidase and IS50R proteins, we find that the regulation of transposition cannot be due to the inhibitor repressing synthesis of Tn5 proteins. Control experiments, in which we assay synthesis of IS50 proteins synthesized from a lambda::IS50R that has been infected into cells carrying the transposition inhibitor, confirm this conclusion. PMID- 6291788 TI - Evidence that lithium and ammonium ions enhance lipopolysaccharide stimulation of lymphoid cells by different mechanisms. PMID- 6291790 TI - [Dietary fiber]. PMID- 6291789 TI - [HBsAg positive cirrhosis of the liver with hepatocellular carcinoma, malignant gastric lymphoma and clear-cell renal adenoma]. PMID- 6291791 TI - Studies on viomycin. XV. Comparative study on the specificities of two anti viomycin antisera by enzyme immunoassay. PMID- 6291792 TI - Bacteriological comparison of the activities of ceftriaxone, a new long-acting cephalosporin, with those of other new cephalosporins. PMID- 6291793 TI - [Diagnosis of cytomegalovirus infections in newborn infants and infants: the value of indirect immunofluorescence compared with complement fixation and virus isolation]. PMID- 6291794 TI - Surveillance of orthopoxvirus infections, and associated research, in the period after smallpox eradication. AB - In 1980, the World Health Assembly declared the global eradication of smallpox and recommended the universal discontinuation of smallpox vaccination; nevertheless, it recommended that surveillance and research on orthopoxvirus infections should continue. By early 1982, all except 8 countries in the world had stopped routine vaccination programmes and all except 1 no longer required an international certificate of smallpox vaccination for travellers. Since 1978, as a result of continuing active surveillance, 176 smallpox rumours have been investigated in 60 countries. Two of these concerned the two laboratory associated cases that occurred in the United Kingdom in 1978; all the others were false alarms. Special surveillance programmes for human monkeypox have been developed in West and Central Africa. The number of laboratories retaining variola virus stocks has been reduced to four. Investigations to determine the identity and origin of the six known isolates of "whitepox" virus have continued. Research on mapping of variola DNA and on monoclonal antibodies against certain orthopoxvirus antigens is continuing. All these measures are aimed at ensuring that the achievement of smallpox eradication is permanent. PMID- 6291795 TI - Laboratory investigation of two "whitepox" viruses and comparison with two variola strains from southern India. AB - Two variola-like viruses were isolated in Bilthoven in 1964 from monkey kidney tissue cultures. These viruses, coded 64/7255 and 64/7275, have been considered as two of the six "whitepox" viruses isolated from animal tissues, all of which are indistinguishable from variola virus by laboratory tests.Two specimens from suspect smallpox cases in India were examined in the Bilthoven laboratory at about the same time as the "whitepox" viruses and two strains of variola virus were isolated from them. A detailed comparison of certain biological markers of these four viruses, and of their DNAs, shows that the two "whitepox" viruses could not be distinguished from each other or from one of the two variola isolates. In view of this, and since there was a possibility of cross contamination at the time of isolation, it is concluded that 64/7255 and 64/7275 must be regarded as genuine variola viruses and be deleted from the list of variola-like viruses isolated from animal tissues. PMID- 6291796 TI - Clinical effectiveness of some fluoride-containing toothpastes. AB - Comparisons were made of the clinical effectiveness of two small groups of fluoride-containing toothpastes on the basis of published and unpublished information available to the group. One comparison showed that particular sodium fluoride/silica and stannous fluoride/calcium pyrophosphate formulations were effective in reducing the incidence of dental caries in schoolchildren and that the former toothpaste was more effective than the latter. A separate comparison showed that certain toothpastes containing sodium monofluorophosphate formulated with either an alumina or an insoluble metaphosphate abrasive were also effective in reducing and controlling caries. The group recommended that there was a need for additional field trials in which direct comparisons could be made between a wide variety of formulations, and that further research should be carried out to develop improved formulations. Extension of the use of adequately formulated fluoride-containing toothpastes is recommended as a valuable public health measure to reduce the incidence of dental caries. PMID- 6291797 TI - Small cell anaplastic carcinoma of the lung. A review of growth characteristics and implications for chemotherapy. AB - Small cell anaplastic lung cancer is increasingly considered to be potentially curable. The opinion that this tumor is a rapidly proliferating one has not been substantiated by kinetic studies, and for this reason late relapse may occur in complete remitters. Differential rates of response/relapse in different tumor sites may be explained on the basis of heterogeneity in tumor kinetics. PMID- 6291798 TI - Neural regulation of cyclic AMP, cyclic AMP-dependent protein kinase, and phosphorylase in bullfrog ventricular myocardium. PMID- 6291800 TI - Neurogenic electrical responses of single smooth muscle cells of the dog middle cerebral artery. AB - Electrical responses induced by perivasascular nerve stimulation were recorded intracellularly from the smooth muscle of dog middle cerebral artery. With nerve stimulation, the muscle membrane produced excitatory junction potential and then a slow hyperpolarization. The excitatory junction potential showed facilitation and the slow hyperpolarization showed depression phenomena, when the nerves were stimulated with twin pulses. Generation of the slow hyperpolarization was associated with an increase in the potassium conductance of the membrane and was suppressed by tetraethylammonium, which depolarized the membrane, reduced the membrane conductance, and increased the amplitude of the excitatory junction potential. Treatment with 6-hydroxydopamine abolished the excitatory junction potential, but not the slow hyperpolarization; the latter was suppressed by tetrodotoxin. The amplitude of slow hyperpolarization was decreased by application of tetraethylammonium or ATP, but was not affected by application of atropine, neostigmine, theophylline, apamin, ouabain, norepinephrine, propranolol, or guanethidine. ATP produced transient depolarization of the membrane with associated decrease in the membrane resistance. The excitatory junction potential was attributed to activation of the noradrenergic nerves, whereas the slow hyperpolarization was not generated by activation of adrenergic, cholinergic, or purinergic receptors. Inasmuch as the electrogenic Na-K pump, cAMP, and ATP were not involved in the generation of slow hyperpolarization, the possibility of an unidentified chemical transmitter should be given attention. PMID- 6291799 TI - Extracellular action of adenosine and the antagonism by aminophylline on the atrioventricular conduction of isolated perfused guinea pig and rat hearts. PMID- 6291801 TI - Regional and systemic metabolic effects of angiotensin-converting enzyme inhibition during exercise in patients with severe heart failure. AB - The acute hemodynamic and metabolic effects of captopril therapy were studied in 12 patients with severe heart failure during maximal exercise performed on an upright bicycle ergometer. During the control period, exhaustion occurred after 4.2 +/- 2.7 minutes of exercise. Cardiac index increased from 1.54 +/- 0.36 l/min/m2 at rest to 3.39 +/- 1.54 l/min/m2 (p less than 0.001) at exhaustion; systemic arteriovenous oxygen difference increased from 8.8 +/- 2.1 to 12.8 +/0 2.4 ml/100 ml (p less than 0.001) and oxygen uptake from 3.4 +/- 0.5 to 10.8 +/- 3.0 ml/kg/min (p less than 0.001). Pulmonary arterial oxygen content decreased from 7.3 +/- 1.3 to 3.7 +/- 1.5 ml/100 ml (p less than 0.001) and femoral vein oxygen content from 5.0 +/- 1.7 to 2.5 +/- 1.2 ml/100 ml (p less than 0.001). During captopril therapy, cardiac index significantly increased both at rest (1.83 +/- 0.54 vs 1.54 +/- 0.36 l/min/m2, p less than 0.01) and during maximal exercise (3.67 +/- 1.51 vs 3.39 +/- 1.54 l/min/m2, p less than 0.01). Systemic arteriovenous oxygen difference decreased significantly at rest, from 8.8 +/- 2.1 to 7.7 +/- 2.1 ml/100 ml (p less than 0.01) and during maximal exercise from 12.8 +/- 2.4 to 12.3 +/- 2.2 ml/100 ml (p less than 0.01). Pulmonary arterial oxygen content at exhaustion was significantly higher during captopril therapy than during the control period (4.1 +/- 1.1 vs 3.7 +/- 1.5 ml/100 ml, p less than 0.05), while femoral venous blood content was unchanged. Captopril therapy did not significantly increase maximal oxygen uptake or exercise duration. Thus, the acute administration of captopril to patients with severe heart failure does not increase exercise capacity despite improved cardiac performance. Moreover, captopril therapy does not acutely result in metabolic benefits to the skeletal muscles during exercise. PMID- 6291802 TI - Possible involvement of an endogenous opioid in the antihypertensive effect of clonidine in patients with essential hypertension. AB - The effect of naloxone on the hypotensive and bradycardiac action of clonidine was studied in 27 hospitalized patients with uncomplicated mild-to-moderate essential hypertension. In a double-blind, crossover study, clonidine, 0.3 mg/day orally for 3 days, significantly reduced systolic and diastolic blood pressure and heart rate, whereas placebo was ineffective. Naloxone, 0.4 mg given intravenously on the third day of clonidine treatment, caused a rapid increase in blood pressure and heart rate in 14 patients (reacting group), but was ineffective in the remaining 13 patients (nonreacting group). Naloxone given during the placebo period was ineffective in all patients. Both the clonidine induced hypotension and the rebound increase in blood pressure after cessation of clonidine were significantly greater in the reacting than in the nonreacting group. These observations suggest that release of an endogenous opioid contributes to the antihypertensive action of clonidine; this mechanism may be also involved in the discontinuation syndrome after cessation of clonidine. PMID- 6291803 TI - The interaction of sodium nitroprusside with human endothelial cells and platelets: nitroprusside and prostacyclin synergistically inhibit platelet function. AB - Sodium nitroprusside (NP) is a potent vasodilator that also inhibits platelet aggregation. To test the hypothesis that NP causes both of these effects by altering the balance between prostacyclin (PGI2) produced by endothelial cells and thromboxane A2 (TXA2) produced by platelets, we incubated each of these cell types with NP for 5 minutes and assayed the PGI2 and TXA2 produced. NP at pharmacologically achieved doses (0.01--30 micrograms/ml) inhibited platelet aggregation and resultant TXA2 synthesis in a dose- and time-dependent manner (p less than 0.001). The inhibition was not dependent on cAMP production, external calcium concentration, or suppression of TXA2 synthesis. NP did not alter the production of PGI2 by cultured human endothelial cells as measured by radioimmunoassay for 6-Keto-PGF1 alpha, the stable hydrolysis product of PGI2. However, supernates of NP-treated endothelial cells containing low, noninhibitory concentrations of NP unexpectedly inhibited platelet aggregation. This inhibition of platelet aggregation was due to synergy between PGI2 (0.1--3 nM) and NP (p interaction less than 0.03). The synergistic inhibition by NP and PGI2 of platelet aggregation and TXA2 synthesis in vivo may explain some of the beneficial actions of NP in the treatment of hypertension and congestive heart failure. PMID- 6291804 TI - Animal model of congestive cardiomyopathy. PMID- 6291805 TI - The acute hemodynamic effects of intravenous verapamil in coronary artery disease. Assessment by equilibrium-gated radionuclide ventriculography. AB - The acute hemodynamic effects of an i.v. bolus of verapamil, 0.1 mg/kg or 0.06 0.075 mg/kg, were examined by serial radionuclide studies in 46 patients with coronary artery disease. In 20 patients with ejection fractions (EFs) greater than 35% (group 1A), verapamil, 0.1 mg/kg given over 1-11/2 minutes, had a biphasic effect: first, a transient decrease in EF accompanied by increased left ventricular (LV) volumes and cardiac output equivalents; then, an overshoot of EF to values above control, accompanied by a decrease in peripheral vascular resistance and a drastic decrease in LV volumes, while cardiac output equivalent remained slightly elevated. In eight patients with EFs less than 35% (group 1B), only the first effect on EF was noted. In 10 patients with EFs greater than 35% (group 2), verapamil, 0.06-0.075 mg/kg, exerted qualitatively similar but milder effects on hemodynamic function. Finally, verapamil, 0.1 mg/kg given more slowly, over 2-21/2 minutes, produced no significant changes in EF or LV volumes in another eight patients (group 3). The acute effects of verapamil are thus both time-related and dose-dependent. They are also related to the baseline functional reserve of the left ventricle. This study documents that verapamil exerts a depressant effect on LV function. However, the transient nature of this depression and the quick recovery to normal or above-normal values indicate that verapamil, in the doses used in this study, is safe to use intravenously in patients with coronary artery disease. PMID- 6291806 TI - Physical training increases ventricular fibrillation thresholds of isolated rat hearts during normoxia, hypoxia and regional ischemia. AB - The effect of exercise training on cardiovascular mortality is controversial. The purpose of this study was to determine the effect of a period of treadmill training on the ventricular fibrillation threshold of the isolated rat heart. Trained hearts had higher threshold values during standard, control perfusion conditions, and when exposed to hypoxia, hypoxia plus isoproterenol infusion, or when subjected to coronary artery ligation. Myocardial metabolic studies failed to define the mechanism for the effect of running training. However, in coronary ligated hearts, the content of the arrhythmogenic substance 3',5' cyclic adenosine monophosphate (cyclic AMP) was reduced in the ischemic zone of hearts from trained rats. Cyclic AMP levels were also lower in trained hearts during control perfusions. We conclude that running training increases the resistance of the heart to ventricular fibrillation by mechanisms that are largely unknown, although they may involve cyclic AMP. PMID- 6291807 TI - Side effects of long-term amiodarone therapy. PMID- 6291808 TI - An improved radioreceptor assay for TSH receptor antibodies. AB - An improved receptor assay for TSH receptor antibodies is described in which detergent-solubilized TSH receptors and 125I-labelled TSH are used. Normal human serum and immunoglobulin concentrates from normal serum showed little effect on the interaction between labelled TSH and detergent-solubilized receptors whereas immunoglobulin concentrates from some Graves sera caused marked inhibition of TSH binding. Precipitation with polyethylene glycol was the most convenient way of preparing immunoglobulin concentrates and using this technique the assay could be completed in a few hours. The coefficient of inter assay variation at 51% inhibition of labelled TSH binding was 3.7% and analysis of serum samples from patients with Graves' disease (n = 42), Hashimoto's disease (n = 26), multinodular goitre (n = 9), rheumatoid arthritis (n = 10) and normal donors (n = 35) suggested that the assay could detect TSH receptor antibodies in about 80% of patients (treated and untreated) with Graves' disease. PMID- 6291809 TI - Role of prostaglandins in the aldosterone response to ACTH in sodium depleted human subjects. AB - It is well established that the response of plasma aldosterone to ACTH is enhanced in the sodium depleted state. The mechanisms for this phenomenon are not clear, however, and the present study was undertaken to determine the possible participation of endogenous prostaglandins. ACTH, 250 ug by i.m. administration, was given to 10 human subjects pretreated in four different ways: 1. Control, receiving a 200 mEq per day sodium diet; 2. Sodium depletion (60 mEq/day sodium plus furosemide) plus indomethacin; 3. Sodium depletion plus indomethacin plus captopril; and 4. Sodium depletion plus captopril. Only in the last group, in which the prostaglandin cyclooxygenase inhibitor, indomethacin, was not given during the sodium depletion, did an exaggerated aldosterone response to ACTH occur (an increase of 468% compared with an increase of 182% during control, P less than 0.005). The angiotensin converting enzyme inhibitor, captopril, did not effect this response. Thus, endogenous prostaglandins appear to be of far greater importance than the renin-angiotensin system in mediating the increased aldosterone response to ACTH administration during the sodium depleted state in man. PMID- 6291810 TI - Effects of angiotensin III and ACTH on aldosterone secretion. AB - ACTH and des-Asp1-angiotensin II (AIII) can raise plasma aldosterone. To assess the threshold for ACTH and AIII stimulated adrenal steroidogenesis we infused ACTH (from 0.03 to 10 ng ACTH/min) and AIII (from 0.1 to 20 ng/kg/min) to dexamethasone pretreated sodium deplete normal subjects and patients with primary aldosteronism, chronic renal failure, and end stage renal disease maintained with continuous ambulatory peritoneal dialysis. Plasma aldosterone in the primary aldosteronism group increased significantly at 0.3 ng ACTH/min compared with 1 to 3 ng ACTH/min in all other groups. The threshold dose for an ACTH stimulated rise in plasma aldosterone was as at least as low as the dose necessary to raise cortisol in all groups. The threshold dose for an AIII stimulated rise in plasma aldosterone was 4 ng/kg/min in normals and between 1 and 3 ng/kg/min in primary aldosteronism. The metabolic clearance rate (MCR) of aldosterone was determined by constant infusion of [3H]-aldosterone. The decline in MCR during AIII infusion contributed less than 15% to the rise in plasma aldosterone in normals and patients with primary aldosteronism. PMID- 6291811 TI - The effect of captopril on renin, angiotensin II, cortisol and aldosterone during ACTH-infusion in man. AB - Prolonged low-dose ACTH infusion (5 or 10 iU/24h) leads to a transient increase in plasma renin activity and angiotensin II concentration in normal man. In order to find out whether the increase in angiotensin II stimulates aldosterone secretion, 12 normal men received ACTH (10 IU/24h) for 34 hours altogether, 6 with and 6 without simultaneous administration of captopril, 50 mg every 6 hours. Captopril prevented the increase in plasma angiotensin II during ACTH infusion and lowered its levels below those on the control day two hours after a new dose of the converting enzyme inhibitor was given. The increase in plasma cortisol was similar in both groups. The increase in plasma aldosterone was significantly blunted by captopril. The early blood pressure rise and the kaliuresis during ACTH infusion were also significantly decreased in the captopril group. These results suggest that angiotensin II mediates in part the effect of ACTH on aldosterone and blood pressure during the first 2 days of infusion. Since captopril reduced plasma angiotensin II for some time below normal, it is alternatively possible that ACTH requires normal plasma angiotensin II levels for a full effect on aldosterone secretion. PMID- 6291812 TI - Conversion of progesterone from the adrenal or ovary to deoxycorticosterone in plasma. AB - The in vivo conversion of progesterone to deoxycorticosterone (DOC) by peripheral 21 hydroxylation was studied in normal men and women. The technique used was to infuse labeled progesterone under steady state conditions and isolate the precursor and product from blood by multiple chromatography steps. DOC was chromatographed as both the steroid and its acetylated derivative. The conversion rate was found to be 1.3 +/- 0.1% in men and women despite differences in the concentration of progesterone during the cycle. Since progesterone was very low in men and women in the follicular phase of the cycle, we conclude that DOC is derived from direct secretion. However, progesterone rises to 1073 +/- 101 ng per dl in the luteal phase and the calculations indicate that DOC increases from 8.3 +/- 0.4 (SE) to 15.6 +/- 1.3 ng per dl. 3/4 of luteal phase DOC is from conversion of progesterone. Acute ACTH increases both progesterone and DOC, and almost all of the DOC in plasma is from direct secretion. However, chronic ACTH excess and adrenal 21 hydroxylase deficiency is associated with a fraction (1/4 1/5) of DOC from progesterone indicating that the peripheral conversion rate is unrelated to the adrenal enzyme defect. PMID- 6291813 TI - Primary aldosteronism: effects of inhibition of ACTH and potassium administration on plasma aldosterone concentration. AB - The relative roles of ACTH, angiotensin and potassium in influencing aldosterone secretion in primary aldosteronism were assessed by direct or indirect means. In untreated patients with primary aldosteronism caused either by adrenal adenoma or hyperplasia plasma aldosterone and cortisol concentrations fluctuated in unison and dexamethasone reduced both hormones markedly. Only when renin-angiotensin system was greatly activated and plasma potassium normalized by medical treatment was dexamethasone less successful in lowering plasma aldosterone concentration. Potassium infusion of 10,20 and 30 mEq/hr in patients with adenoma failed to elicit any increase in plasma aldosterone concentration despite significant increases in plasma potassium levels. These results suggest that patients with primary aldosteronism due to adrenal adenoma are relatively more sensitive to small changes in plasma ACTH level than those in plasma angiotensin or potassium levels. In recumbent patients with adrenal hyperplasia ACTH also modulates plasma aldosterone concentration. PMID- 6291814 TI - Regulation of the mineralocorticoid hormones in adrenocortical disorders with adrenocorticotropin excess. AB - Chronic stimulation by adrenocorticotropin (ACTH) of the adrenal cortex produces different plasma mineralocorticoid hormone (MCH) patterns, depending on the amount of glucocorticoid hormones (cortisol) concurrently generated and the degree of activation of the renin angiotensin system (RAS). Patients with Cushing's disease or the ectopic ACTH-excess syndrome have normal or low production of the MCHs, aldosterone and 18-hydroxycorticosterone (18-OHB), by the zona glomerulosa (ZG), elevated cortisol and deoxycorticosterone (DOC) levels, and high-normal to elevated production of the MCHs corticosterone (B) and 18 hydroxydeoxycorticosterone (18-OHDOC) by the zona fasciculata (ZF). Prolonged administration of superphysiologic doses of ACTH to normal subjects yields similar patterns. Patients with simple virilizing 21-hydroxylase deficiency (21 OHD) have impaired ZF production of B and 18-OHDOC and elevated DOC, 18-OHB, and aldosterone secretion secondary to the superimposed RAS stimulation of the ZG. Patients with 17 alpha-hydroxylase deficiency (17 alpha-OHD) have elevated levels of the ZF MCHs DOC, B, 18-OHDOC, and 18-OHB and a functionally suppressed ZG. Patients with 11 beta-hydroxylase deficiency (11 beta-OHD) have only elevated production of DOC by the ZF and suppressed RAS and aldosterone. A significant negative correlation between cortisol and aldosterone concentrations suggests that cortisol is involved in the ACTH-mediated inhibition of aldosterone formation. PMID- 6291815 TI - Increased excretion of 18-hydroxycorticosterone in patients with adrenal adenomas and hypertension. AB - Two female patients, 54 and 34 years old, each presented with an adrenal adenoma and hypertension. Blood pressure fell after removal of the tumors. The first patient had high urinary 18-hydroxycorticosterone and periodically elevated 18 hydroxy-deoxycorticosterone excretions. The second patient had elevated 18 hydroxycorticosterone and free cortisol excretions. Urinary aldosterone, aldosterone metabolites and plasma aldosterone were not increased. Plasma renin activity was suppressed and serum potassium levels were normal. After surgery, no elevated steroid values were found. Elevated 18-hydroxycorticosterone excretion may be an indicator of yet unknown hypertensinogenic mechanisms. The role of 18 hydroxycorticosterone in the etiology of hypertension is still unknown. PMID- 6291816 TI - Circadian secretion of ACTH, cortisol, and mineralocorticoids in Cushing's syndrome. AB - The behaviour of plasma levels of ACTH was studied in five untreated patients with pituitary-dependent Cushing's syndrome, with blood samples taken every half an hour for a total period of 24 hours; plasma cortisol, corticosterone, deoxycorticosterone (DOC) and aldosterone were also measured simultaneously. In all cases, above normal secretory impulses of ACTH and cortisol, at approximately the same height, were present throughout the day, while between these peaks the levels were in the normal range. Few peaks of ACTH and cortisol were simultaneous and rare secretory impulses of corticosterone, and deoxycorticosterone were in synchrony with those of ACTH or cortisol. DOC levels were found to have some peaks above normal levels while corticosterone levels presented rare elevated peaks during the day. Plasma aldosterone values on the other hand, were extremely low in all except in one case, where the variations may be interpreted as pure fluctuations. These findings confirm that: 1) In a number of cases, multiple samples of ACTH and cortisol (during the 24 hrs.) appear to be essential in order to distinguish pituitary-dependent Cushing's syndrome from normal; 2) In conditions of ACTH excess, DOC and corticosterone secretion seems to become progressively less ACTH-dependent as we proceed down the biosynthetic chain towards aldosterone, which is suppressed in most cases. PMID- 6291818 TI - 19-Nor-corticosteroids in experimental and human hypertension. AB - Recent reports demonstrate that the 19-nor-corticosteroids (19-nor-DOC) are naturally-occurring substances in hypertensive animal models as well as man. Since some 19-nor-corticosteroids are potent mineralocorticoids, they may have a role in regulating systemic arterial pressure and be involved in the pathogenesis of hypertension. This paper reports the probable biosynthetic pathway, factors regulating the secretion or production, and measurement of 19-nor-DOC in man and the spontaneously hypertensive rat (SHR). These studies demonstrate (1) 19-nor DOC is greatly influenced by ACTH and dexamethasone but less so by high and low salt diets in normotensive subjects; (2) 19-nor-DOC is greatly increased in some but not all hypertensive patients; (3) 19-nor-DOC is increased in prehypertensive SHR compared to WKY rats. The likelihood of metacorticoid hypertension and possible role of other 19-nor-corticosteroids, including 19-nor-progesterone, are discussed. It can be concluded that 19-nor-corticosteroids are synthesized by extra-adrenal tissues in biologically active quantities. They are increased and possibly pathogenetic in certain states of human and experimental hypertension. PMID- 6291817 TI - Mechanism of the response to captopril in glucocorticoid hypertension. AB - The effect of captopril was explored in salt-depleted methylprednisolone (MP) hypertensive rats. MP treatment raised BP by 41+/-2 mmHg over 2 weeks. Controls (C) had no change in BP. Sodium balance and weight data indicated a greater salt depletion in MP than in C. On day 15, captopril reduced BP in both MP (20+/-4 mmHg) and C (31+/-4 mmHg). The effect was significantly smaller in MP than in C (p less than 0.05). Plasma renin activity (PRA) was similarly elevated in both groups, consistent with salt depletion. Serum (SCE) and lung-converting enzyme (LCE) activity were similar in MP and C. The diminished antihypertensive effect of captopril in MP is therefore not attributable to differences in PRA, SCE, or LCE. Our data suggest that depressor actions of captopril unrelated to the renin angiotensin system are impaired in MP. Glucocorticoid-induced changes in vasodilator systems may explain these findings. PMID- 6291819 TI - Aldosterone response to antihypertensive treatment. AB - To investigate whether changes in aldosterone during antihypertensive treatment would be related to alterations in the renin-angiotensin system or to changes in sodium-fluid balance, 54 essential hypertensives were hospitalized. Sodium intake was restricted to 55 mmoles per day. Levels of renin, angiotensin II and aldosterone were measured before and after two weeks treatment with atenolol (n=15), prazosin (n=15), the converting enzyme inhibitor MK 421 (n=6), verapamil (n=9) and the vasodilator L 6150 (n=10). Daily sodium excretion was determined from 24 h urine collections. The results indicate that, when renin or angiotensin levels do not change, the aldosterone response depends on alterations in sodium balance. When the renin system is depressed, sodium loss may prevent a large drop in aldosterone levels. It is concluded that during antihypertensive treatment body sodium status in itself modifies aldosterone secretion, irrespective of the renin-angiotensin system. PMID- 6291820 TI - [Ectopic hormone producing tumors]. PMID- 6291821 TI - [Mechanisms of hypoglycemia associated with extrapancreatic tumors]. PMID- 6291822 TI - Food and diabetes: the dietary treatment of insulin-dependent and non-insulin dependent diabetes. PMID- 6291823 TI - Diverticular disease--is it a motility disorder? PMID- 6291824 TI - HLA and hormonal data for identification of heterozygotes in 11 beta- and 17 alpha-hydroxylase deficiency syndromes. AB - 1. In previous studies, baseline and ACTH-stimulated hormone levels, plus HLA genotyping, have been used to detect heterozygous carriers in congenital adrenal hyperplasia (CAH) due to 21-hydroxylase deficiency (21-OHDS). 2. In the present study similar parameters were determined in a family of four including two children with CAH due to 11 beta-hydroxylase deficiency (11-OHDS), and a family of twelve including three sibs (two females, one genotypically male) with CAH due to 17 alpha-hydroxylase deficiency (17-OHDS). 3. HLA typing showed affected sibs with 11-OHDS to differ in one of their haplotypes. No significant differences in basal and ACTH-stimulated steroid levels were seen between the parents (obligate heterozygotes) and the general population. 4. In 17-OHDS, affected members differed from one another in one to two haplotypes; one patient had identical HLA profiles with two of the normal siblings, as did the genotypically male patient with two others; each of the other healthy siblings had one haplotype found in two of the affected subjects. The genes responsible for 11-OHDS and 17-OHDS--in contrast with 21-OHDS--do not appear to be HLA-linked. However, the measurement of ACTH-stimulated corticosterone levels may be useful, since the gene responsible for 17-OHDS seems to be expressed hormonally in the heterozygous state. PMID- 6291826 TI - Central alpha-adrenoceptors and blood pressure regulation in the rat. PMID- 6291825 TI - Endogenous and exogenous agonist regulation of responses to beta-adrenoceptor stimulation in patients with chronic autonomic failure. AB - 1. Two patients with orthostatic hypotension due to peripheral autonomic neuropathy were studied. The diagnosis was based on severe hypotension during 60 degrees head-up tilting and absence of the systolic pressure overshoot in phase IV of the Valsalva response. Plasma noradrenaline levels were 50 and 90 pg/ml in the two patients. Noradrenaline was unresponsive to head-up tilting. Heart rate did not change after atropine, 1 mg i.v. Thus the patients had combined efferent sympathetic and parasympathetic lesions. 2. Infusion of isoprenaline in these patients showed supersensitivity for the chronotropic and vascular effects of this drug. Similar cardiac and vascular supersensitivity was observed for salbutamol. The patients were treated with pindolol, 15 mg daily divided in three doses. After 4 weeks treatment pindolol was stopped and the infusion of isoprenaline and salbutamol were repeated 72 h after the last dose of pindolol, 5 mg. The isoprenaline dose-heart rate response curve was shifted to the right by a factor of 9 and 14 in the two patients. The salbutamol dose-heart rate response curve was shifted by a factor of 61 in both patients. 3. The observed chronotropic effects of salbutamol provides evidence for the existance of cardiac beta 2-adrenoceptors in man. Comparison between the cardiovascular responses to the non-selective beta-adrenoceptor agonist isoprenaline and the beta 2-selective agonist salbutamol suggest preferential beta 2-adrenoceptor sensitization in chronic autonomic failure, which can be reversed by treatment with the non selective partial beta-adrenoceptor agonist pindolol. PMID- 6291827 TI - Effects of converting enzyme inhibition with captopril on renal function in normal and ACTH treated sheep. AB - 1. The effect on renal function in sheep of inhibiting converting enzyme with captopril was examined before and after 5 days ACTH administration. 2. Glomerular filtration rate, effective renal plasma flow, effective renal blood flow, mean arterial pressure and plasma sodium were all significantly increased by ACTH treatment and plasma potassium was decreased. Captopril (20 mg i.v.) had no effect on renal function or blood pressure before or after ACTH treatment, although urinary potassium excretion decreased following captopril on day 6 of ACTH treatment. 3. The increase in glomerular filtration rate and effective renal plasma flow seen with ACTH treatment in sheep does not appear to be mediated by the reninangiotensin system. PMID- 6291828 TI - Symposium on Angiotensin Converting Enzyme Inhibition. Report of a meeting held in conjunction with the Third annual meeting of the High Blood Pressure Research Council of Australia, Melbourne, December, 1981. PMID- 6291829 TI - Clinical experience with the angiotensin converting enzyme inhibitor captopril. AB - 1. Sixty-two patients (nineteen mild-moderate, forty-three severe hypertension) were treated with captopril. 2. Blood pressure was controlled long term (with/without diuretics/beta-adrenoreceptor blocking drugs) in sixteen out of nineteen patients with mild-moderate hypertension. Two patients complained of dysguesia. 3. Long term control (with/without other drugs) was achieved in thirty out of forty-three patients with severe or accelerated hypertension, many of whom had associated renal or renovascular disease. Side-effects were common: five rash, five dysguesia, two rash and dysguesia, two reversible agranulocytosis. 4. Six patients had a profound pressure fall after an initial dose of captopril 25 mg and one developed acute oliguric renal failure. Treatment should be commenced at low doses in severely hypertensive patients. PMID- 6291830 TI - Glucocorticoid induction of angiotensin converting enzyme production from bovine endothelial cells in culture and rat lung in vivo. AB - 1. Endothelial cells from bovine aortae, grown to confluence in culture and then maintained for 2 days in serum-free medium produced an enzyme which closely resembled angiotensin converting enzyme (ACE). 2. Dexamethasone increased cell ACE activity 6- to 7-fold with a threshold near 0.3 nmol/l. This effect was completely inhibited by actinomycin D or cycloheximide. 3. Deoxycorticosterone (DOC) and aldosterone were approximately 0.1% as active as dexamethasone. 4. In cells incubated in the presence of dexamethasone (10 nmol/l), DOC (10 mumol/l) inhibited ACE production suggesting that DOC is a partial glucocorticoid agonist/antagonist in this system. 5. Adrenalectomized rats had lower pulmonary ACE compared to intact controls. This was restored by injections of dexamethasone (40 micrograms/day) but not by DOC (40 micrograms/day) or aldosterone (10 micrograms/day). The dose response curve for dexamethasone induction of pulmonary ACE mirrored that for thymolysis; for both, half maximal effects were seen at approximately equal to 6 micrograms/day and plateau levels at approximately equal to 60 micrograms/day. 6. Glucocorticoids appear to be potent inducers of ACE activity in endothelial cells in culture and rat lung in vivo. The action of DOC and aldosterone probably reflects occupancy of glucocorticoid receptors. This effect may be of significance in modulating ACE in local vascular beds. PMID- 6291831 TI - Beneficial haemodynamic effect of the angiotensin blocking agent captopril in patients with refractory heart failure. PMID- 6291832 TI - Correlation between angiotensin converting enzyme inhibition and the acute hypotensive response to MK 421 in essential hypertension. AB - 1. The acute hormonal and hypotensive effects of MK 421 (10 mg p.o.) were assessed by a double-blind randomized trial in twelve subjects with essential hypertension. The study was performed during a normal and low salt intake. 2. Plasma angiotensin converting enzyme (ACE) activity was maximally inhibited by 57 (s.e.m. = 4)% of control activity at 4-8 h. Plasma ACE was still depressed at 24 h by 10% of control activity but had returned to normal by 32 h. 3. Sodium depletion did not alter basal plasma ACE activity, nor change the degree of inhibition of MK 421. 4. Plasma angiotensin II was depressed during ACE inhibition, but no change in circulating bradykinin was detected. Reciprocal rises occurred in plasma renin and angiotensin I levels with a time course similar to ACE inhibition. 5. All subjects showed a fall in their blood pressure, maximal between 6-8 h. Mean arterial pressure fell 12 (s.e.m. = 5) mmHg on the normal sale intake and 16 (s.e.m. = 3) mmHg on the low salt diet, at 8 h. 6. Blood pressure and angiotensin converting enzyme activity changes were significantly correlated (r = 0.86, n = 12). PMID- 6291833 TI - Radical surgical treatment for Krukenberg tumour. PMID- 6291834 TI - The "false-negative" Meckel's scan. AB - A case is presented of a 17-month-old girl who underwent two Meckel's scans with Tc-99m pertechnetate. The initial study was interpreted as normal while a subsequent study five days later was definitely positive. Surgery immediately following the positive Meckel's scan demonstrated a Meckel's diverticulum containing gastric mucosa without evidence of active hemorrhage. This prompted a review of the literature in reference to false-negative Meckel's scans which revealed a wide variance in the reported incidence of false-negative examinations. Repeat scintigraphy in the face of a strong clinical suspicion after an initial normal study may decrease the indicence of false-negative imaging series. PMID- 6291835 TI - The uterine blush. A potential false-positive in Meckel's scan interpretation. AB - To determine the presence, prevalence, and clinical importance of Tc-99 pertechnetate uterine uptake, this retrospective analysis of 71 Meckel's scans was undertaken. Specifically, each study was evaluated for the presence of a focal accumulation of radiotracer cephalad to the bladder. Patients received an intravenous dose of 150 microCi/kg of Tc-99 pertechnetate. Each study consisted of 15 one minute anterior serial gamma camera images, and a 15, 30, and 60 minute anterior, right lateral and posterior scintiscan. Menstrual histories were obtained from all patients except two. No males (33/33), nor premenstrual (13/13), menopausal (4/4) or posthysterectomy (2/2) patients revealed a uterine blush. Eleven of 15 patients (73%) with regular menses demonstrated a uterine blush. They were in the menstrual or secretory phases of their cycle. Four demonstrated no uterine uptake, had regular periods, but were in the proliferative phase of their cycle. Two with irregular periods, and one with no recorded menstrual history, manifested the blush. Radiotracer should be expected in the uterus during the menstrual and secretory phases of the menstrual cycle. It is a manifestation of a normal physiologic phenomenon, and must be recognized to prevent false-positive Meckel's scan interpretations. PMID- 6291836 TI - "Doughnut" technetium pyrophosphate myocardial scintigrams. A marker of severe left ventricular dysfunction. AB - The "doughnut" pattern on Tc-99m pyrophosphate (PPi) myocardial scintigraphy is characterized by a border of tracer uptake surrounding a central zone of relatively decreased activity. This pattern is generally associated with large transmural anterior myocardial infarcts (MI) caused by occlusion or critical stenosis of the left anterior descending coronary artery. Such infarcts typically involve a significant portion of the anterior wall and are associated with a complicated clinical course and poor prognosis. In order to evaluate the relationship between the presence of the doughnut pattern and left ventricular (LV) function, radionuclide ventriculography was performed within 15 days after infarction in 58 patients with transmural anterior MI. In patients without previous MI, 15/38 (39.5%) had doughnut scintigrams. These patients demonstrated significant reductions in LV ejection fraction (EF) (28 +/- 10% versus 45 +/- 12%, P less than 0.001) and normalized LV wall motion scores (29 +/- 11% versus 61 +/- 10%, P less than 0.001) when compared with patients with "nondoughnut" scintigrams. Patients with doughnut scintigrams had a significantly greater incidence of severe septal hypokinesis (P less than 0.001) and apical dyskinesis (P less than 0.03). LV end-systolic volumes were also larger in the patients with doughnut scintigrams (73 +/- 32 ml versus 40 +/- 17 mI/M2, P less than 0.005). In contrast, there was no significant difference in LVEF, normalized LV wall motion score, or LV volumes between doughnut and nondoughnut groups in patients with previous MI. PMID- 6291837 TI - Ectopic gastric mucosa (Meckel's) scintigraphy. PMID- 6291838 TI - Lack of interaction between tricyclic antidepressants and clonidine at the alpha 2-adrenoceptor on human platelets. AB - The pharmacologic interaction between tricyclic antidepressants and clonidine at the alpha 2-adrenoceptor was examined in human platelets by quantifying the ability of tricyclic antidepressant drugs to inhibit clonidine-stimulated platelet aggregation in vitro. Platelet aggregation induced by increasing concentrations of clonidine (0.3 to 3 microM) was not altered by pretreatment of the platelets with 10 microM imipramine. Imipramine at concentrations above 100 microM attenuated clonidine-induced platelet aggregation, but this was a nonspecific drug effect because the high concentrations of imipramine inhibited adenosine diphosphate-induced platelet aggregation as well. Desmethyldoxepin and nortriptyline also inhibited platelet aggregation nonspecifically at higher concentrations (greater than 10 microM). We were also not able to establish a specific interaction between alpha-methlnorepinephrine (the active metabolite of methyldopa) and the tricyclic antidepressants at the platelet alpha 2 adrenoceptor. Our data suggest that if there is an adverse dynamic interaction between tricyclic antidepressants and clonidine, the interaction occurs at a site other than the alpha 2-adrenoceptor. PMID- 6291839 TI - Effects of cimetidine and ranitidine on steady-state propranolol kinetics and dynamics. AB - The influence of cimetidine (1000 mg daily, one day oral pretreatment) and ranitidine (300 mg daily by mouth, 1 and 6 days pretreatment) on steady-state propranolol (160 mg sustained-release capsule, once daily) plasma levels (Psss) and dynamic beta-blocker effects was assessed by bicycle ergometer exercise and isoproterenol sensitivity test in five normal subjects. During the 3 hr of the dynamic tests Psss were elevated from 25% to 67% by cimetidine, whereas Pss was unchanged by ranitidine. The estimated hepatic blood flow (EHBF) as calculated from indocyanine green (ICG) plasma clearance was only slightly reduced by 15 +/- 23% (mean +/- SD, n = 4) after one oral dose of 150 mg ranitidine and showed substantial intersubject variability. Dynamic parameters, like propranolol induced heart rate and blood pressure changes under physical exercise or during the isoproterenol sensitivity test, were not influenced by ranitidine or cimetidine. Since our study was performed on normal subjects with relatively low propranolol doses these results do not rule out the risk of severe reinforcement of beta-adrenergic receptor blocking effects if propranolol and cimetidine are taken together by patients. PMID- 6291840 TI - Skeletal effects of orthovoltage and megavoltage therapy following treatment of nephroblastoma. AB - A review of 19 patients treated for nephroblastoma between 1945 and 1978 by nephrectomy and radiotherapy with chemotherapy showed a marked improvement in cure rate in recent years. In the decade from 1950 to 1960 there are four survivors. From 1960 to 1970, there are nine survivors from 25 cases and since 1970 there are 19 survivors from 27 cases. Skeletal effects of radiation therapy have been reviewed in 22 patients--seven of whom had orthovoltage and 15 megavoltage therapy. Similar skeletal changes followed both orthovoltage and megavoltage therapy, being more severe after the adolescent growth spurt and when the follow-up was longer. The radiation dose was normally 3000 cGy and skeletal changes followed the lowest dose of 2200 cGY. Scoliosis of the lumbar spine, concave to the side of the tumour, with angulation ranging from less than 5 degrees to 54 degrees developed in 12 patients; all but one showed asymmetry of the body LV2. Kyphosis was present in three patients; anterior beaking of the vertebral body in 10; and 16 patients showed asymmetry of more than one vertebral body. Iliac hypoplasia was present in 19 patients, being unilateral when only the renal bed was irradiated. Analysis of the radiological changes expressed as a total score against the age at follow-up shows that severe changes follow orthovoltage therapy. It is too early to assess the full extent of megavoltage therapy simply because patients have not been followed up for a comparable length of time. PMID- 6291841 TI - Stimulation of beta-adrenoceptors in the exercising human forearm. AB - In order to study the effects of local stimulation of beta-adrenoceptors on limb blood flow during exercise and on muscle metabolism, the non-selective beta adrenoceptor agonist, isoprenaline, was infused intra-arterially into the forearm of six healthy, young subjects. Isoprenaline caused a marked increase in forearm blood flow not only at rest but also during dynamic forearm exercise. The forearm release of lactate increased considerably during drug infusion whereas glucose uptake decreased. The net uptake of free fatty acids during exercise decreased. There were no systemic effects of isoprenaline. These findings suggest that the resistance vessels are responsive to beta-adrenergic vasodilatation also during muscle exercise. The increase in blood flow through the exercising forearm may be secondary to an increased muscle metabolism. The increased lactate release from the exercising muscles indicates an increased muscle glycogen breakdown during beta-adrenoceptor stimulation. PMID- 6291843 TI - [Pharmacological suppression of the adrenal gland. Considerations on a clinical case]. PMID- 6291842 TI - [The physiopathology of opioid peptides]. PMID- 6291844 TI - [Clinical and instrumental evaluation of patients with chronic peripheral arterial occlusive disease after prolonged administration of suloctidil]. PMID- 6291845 TI - [Implantation of Rous sarcoma virus transformed cells in the hamster: effect of various parameters]. AB - The effect of inoculation of allogeneic hamster cells transformed by Rous sarcoma virus on tumour growth, in the hamster is described. Inoculation of low doses (10(2)-10(4)) of live cells was followed by the apparition and the growth of sub cutaneous sarcomas. Repeated injections of irradiated or glutaraldehyde-treated cells which induce immune reactions led only to partial protection since they did not totally prevent tumour growth, after challenge with strong doses (5 X 10(4)) of live transformed cells. Inoculation route did not influence tumour growth, whatever the nature of injected cells. The interest of using irradiated or glutaraldehyde-treated transformed cells in tumour immunity should be emphasised. PMID- 6291846 TI - Studies on the role of humoral and cell-mediated immunity in pigs after vaccination with Aujeszky's disease virus. AB - Humoral and cellular immunity in pigs vaccinated twice with Aujeszky's disease virus (ADV) was studied by seroneutralizing test and direct leucocyte migration inhibition technique. Significant migration inhibition of leucocytes (LMI) was found on the fifth day, whereas specific antibodies began to appear at that time only in very low titers. Anamnestic reaction due to the second injection of ADV did not bring about a significant increase of migration inhibition of leucocytes, instead the level of antibodies elevated markedly. PMID- 6291847 TI - Extension of recurrent rectal carcinoma through sciatic foramen: diagnosis by computed tomography. PMID- 6291848 TI - The role of computed tomography in a case of an ACTH secreting thymic carcinoid tumour. PMID- 6291849 TI - Collagen degradation. PMID- 6291850 TI - Placental morphology of low-birth-weight infants born at term. AB - Gross and microscopic examination in 50 placentas of low-birth-weight infants showed: (1) hematogenous infection (74%); (2) placental circulatory disturbances related to maternal hypertension (14%); (3) abnormal placentation (6%); (4) isolated villous dismaturity (4%), and (5) diffuse chorioangiomatosis (2%). As the main placental lesion associated to low-birth-weight in this series was hematogenous infection, the author stress the validity of the virologic, bacteriologic and parasitologic examination of the placenta combined with the morphologic one in the detection of the etiology of intrauterine infection. PMID- 6291851 TI - Placentitis. PMID- 6291852 TI - Does an endogenous digoxin-like immunoreactive factor participate in the development of cardiomegaly? PMID- 6291853 TI - Computed tomography in bone and soft tissue neoplasm: application and pathologic correlation. PMID- 6291854 TI - ADP-ribosylation reactions. PMID- 6291855 TI - The role or urea synthesis in the removal of metabolic bicarbonate and the regulation of blood pH. PMID- 6291856 TI - The role of phosphorylation in the regulation of eukaryotic initiation factor 2 activity. PMID- 6291858 TI - From cat to cricket: the genesis of response selectivity of interneurons. PMID- 6291857 TI - Criteria for establishment of the biological significance of ribosomal protein phosphorylation. PMID- 6291859 TI - The neuroembryological study of behavior: progress, problems, perspectives. PMID- 6291860 TI - Development, maintenance, and modulation of patterned membrane topography: models based on the acetylcholine receptor. PMID- 6291861 TI - Tinea versicolor: treatment and prophylaxis with ketoconazole. PMID- 6291862 TI - Strategies for detecting and characterizing restriction fragment length polymorphisms (RFLP's). PMID- 6291863 TI - Suppression of transformation and tumorigenicity in interspecies hybrids of human SV40-transformed and mouse 3T3 cell lines. AB - Somatic cell hybrids, formed by fusion of human SV40-transformed fibroblast lines and mouse 3T3 cells, were isolated and analysed for expression of transformation phenotypes and tumorigenic potential in immunodeficient nude mice. Both tumorigenic and nontumorigenic SV40-transformants were used for these experiments. Regardless of whether or not the parental human transformed parent line was tumorigenic, the hybrid progeny-with rare exception-did not form tumors. The possibility that human SV40-transformed lines and the derived hybrid progeny were immunologically rejected in nude mice was unlikely since fusion of these human lines to a tumorigenic 3T3 variant produced hybrids which were highly tumorigenic. In addition, in these hybrid lines, there was not a coordinate expression of SV40 T-antigen and transformation traits. The phenotypic separation in somatic cell hybrids of SV40 T-antigen expression and transformation phenotypes and tumorigenicity is evidence that host genetic changes, occurring during or subsequent to viral integration, also contribute to the expression of those traits in SV40-transformed cells. The general inability of human SV40 transformations to form tumors in nude mice would result from their inherent nontumorigenicity and reflect the relative rarity, compared to SV40-transformed mouse cells, of those host genetic changes determining tumorigenicity. The postulated role of an interaction between viral and cellular genetic changes was supported by other results hybrid lines formed by fusion of LNSV cells and the tumorigenic 3T3 derivative were markedly more anchorage independent than either parental line or LNSV/3T3 hybrid cells. PMID- 6291864 TI - Assessment of activity in Sarcoidosis. Sensitivity and specificity of 67Gallium scintigraphy, serum ACE levels, chest roentgenography, and blood lymphocyte subpopulations. AB - The value of different factors are examined to assess activity in 60 patients with biopsy-proven sarcoidosis. In patients with active sarcoidosis (n = 35), 67Gallium scans proved to be the most sensitive method (94 percent sensitivity), followed by serum angiotensin I converting enzyme (S-ACE) levels, chest x-ray films, and lymphocyte assays. In patients with peripheral pulmonary lesions, chest x-ray films failed in 32 percent of cases to document activity (68 percent sensitivity) whereas 67Ga scans and S-ACE levels remained to give reliable results. Despite poor specificity, negative 67Ga scans together with normal ACE levels have a high predictive value for exclusion of active sarcoidosis. In patients with peripheral pulmonary lesions, chest roentgenography is of doubtful value for staging lung involvement and assessment of activity including monitoring and control of therapy. PMID- 6291865 TI - Study of the diffusion of cefotiam in the bronchial secretions. AB - The objective of this study was to evaluate the concentration in the bronchial secretions of a new injectable cephalosporin, cefotiam, which is characterized by an excellent antibacterial activity, stability against beta-lactamases and interesting pharmacokinetic properties. 30 patients suffering from acute exacerbation of a chronic bronchitis or bronchiectasis with expectoration, received either 1 g i. v. of cefotiam in a single dose or four doses of 1 g each over a period of 3 days. The bronchial secretion samples were obtained by fibroscopy, 1, 2, or 4 h after the last dose, in order to evaluate the kinetics of the bronchial concentration of cefotiam. Blood samples were taken simultaneously in order to establish a possible correlation between the bronchial and serum levels of the drug. The results of the study show a rapid and significant transfer of the drug through the bronchoalveolar capillaries, elevated bronchial concentrations starting from the 2nd h, reaching 10 micrograms/ml (or more) in certain cases, slow elimination with bronchial concentrations persisting at high levels for up to 4 h and a ratio between bronchial and serum concentrations of 25-60% at the 2nd h and more than 100% at the 3rd and 4th h. These results confirm the excellent diffusion of cefotiam, especially in the respiratory tract, allowing bronchial levels largely superior to the minimum inhibitory concentrations required to kill the bacteria usually responsible for lower respiratory tract infections. PMID- 6291866 TI - Simultaneous simulation of the serum profiles of two antibiotics and analysis of the combined effect against a culture of Pseudomonas aeruginosa. AB - The serum profiles of ceftriaxone (CEF) and netilmicin (NET) were produced in a liquid culture of Pseudomonas aeruginosa, and the change in viable count recorded. Synergism was detectable between CEF and NET despite of the widely disparate serum halflife times of NET and CEF. PMID- 6291867 TI - Polymyxin B-induced cocarde growth phenomenon of Serratia marcescens due to cationic detergent-like activity of polymyxin B. AB - 14 of 74 test strains of Serratia marcescens yielded reproducible cocarde-like growths (coc+) around 30-micrograms disks of polymyxin B (PB) on Muller-Hinton, brain heart infusion and tryptic soy agar. The coc+ phenomenon was not due to nutrient effects of growth medium nor did it correlate with either group A (phage tail) bacteriocinogeny or colicinogeny as determined with 32 selected test strains; mitomycin C failed to give rise to coc+ growths. The anionic bile salts of MacConkey agar as well as aqueous sodium deoxycholate neutralized the coc+ activity of PB. Benzalkonium chloride, chlorhexidine digluconate, and cetyltrimethylammonium bromide by themselves did not produce cocardes. Rather, these cationic detergents enhanced PB activity somewhat against selected coc+ and coc- strains of S. marcescens. It was concluded that the PB-induced growth phenomenon of S. marcescens was due to the cationic detergent-like activity of this polypeptide antibiotic. PMID- 6291868 TI - Effect of ceftriaxone on Pseudomonas aeruginosa and Staphylococcus aureus in broth, serum, and in combination with human polymorphonuclear leukocytes. AB - We investigated the antibacterial activity of ceftriaxone at concentrations of 1/4 X minimum inhibition concentration (MIC), 1 X MIC and 4 X MIC against a serum resistant Pseudomonas aeruginosa and a serum-resistant STaphylococcus aureus strain in broth, serum, and in combination with leukocytes. Killing effect of ceftriaxone in broth was significantly better than in serum; ceftriaxone improved leukocyte bactericidal activity without serum on P. aeruginosa, but not on S. aureus. The antibacterial activity of ceftriaxone was most effective in combination with leukocytes and serum, achieving a marked bactericidal effect already at subinhibitory ceftriaxone concentrations. PMID- 6291870 TI - Treatment of experimental ascending Escherichia coli pyelonephritis with ceftriaxone alone and in combination with gentamicin. AB - We have tested the effectiveness of several antibiotic regimens, using a rat model of Escherichia coli experimental pyelonephritis that mimics the conditions of severe renal infections in man because the infection is acquired by the ascending route. We found that ceftriaxone, when given for 5 days to rats with severe exudative pyelonephritis, was as effective as the combination ceftriaxone + gentamicin or the reference combination ampicillin + gentamicin. This effectiveness in vivo of the antibiotic alone was achieved despite a marked synergism between the combinations of antibiotics in vitro. This observation suggests that a new and extremely active cephalosporin is as effective in vivo when used alone as when given in combination with an aminoglucoside and provides rationale for testing the use of single antibiotic therapy for clinical situations for which combinations of antibiotics are currently recommended. PMID- 6291869 TI - Combination effect of ceftriaxone with four aminoglycosides on nonfermenting gram negative bacteria. AB - The in vitro efficacy of ceftriaxone in combination with gentamicin, tobramycin, amikacin and netilmicin against 50 nonfermenting gram-negative bacterial strains was compared by use of the checkerboard agar dilution technique. On average 42.5% of all nonfermenting strains were inhibited by additive, 22.5% by synergistic ceftriaxone-aminoglycoside combinations. Great variations occurred between the different bacterial species. Ceftriaxone-tobramycin interactions were superior to combinations with other aminoglycosides. Ceftriaxone-aminoglycoside combinations were most active on Pseudomonas aeruginosa and least potent on Pseudomonas cepacia. PMID- 6291871 TI - Heterotopic arrhenoblastoma. A case report. PMID- 6291872 TI - CSF cyclic AMP changes in nervous system inflammatory diseases. PMID- 6291874 TI - Leukocyte migration-inhibition agarose test in swine after vaccination and challenge with inactivated and activated pseudorabies virus. AB - Cell-mediated immunity to pseudorabies virus (PrV) was studied by means of the leukocyte migration-inhibition (LMI) agarose test. Migration of leukocytes from swine vaccinated with 10 TCID50 inactivate PrV and from normal control swine was not inhibited in the presence of specific PrV antigen. Cellular immune responses were detected in 80% of the vaccinated and 40% of the control group animals at the 1st week after challenge exposure. At the 2nd week, both groups of swine showed a peak level (p less than 0.001) immune response in the LMI test. This level of response remained until the 3rd week in the vaccinated group and the 2nd week in the control group. The averages of migration indices at the 1st, 2nd, 3rd and 4th weeks after challenge exposure were 0.74, 0.22, 0.60 and 0.89 in the vaccinated and 0.82, 0.29, 1.01 and 1.12 in the control group, respectively. Low concentrations of virus-neutralizing (VN) antibodies were present in 30% of the vaccinated swine at the 1st week, and 60% at the 2nd week after vaccination. Anamnestic antibody and primary VN antibody responses occurred after challenge in vaccinated and normal swine, respectively. PMID- 6291873 TI - Experimental investigations on canine endotoxic shock: changes in hemodynamics and involvement of some humoral factors. PMID- 6291875 TI - [A new human hepatoma cell line: establishment and characterization]. AB - We have grown a human hepatoma cell line, designated as HA22T/VGH, from a 52-yr old male hepatoma patient since July 1, 1980. This cell line has been subcultured more than 100 passages. The chromosome analysis of HA22T/VGH indicated that the chromosome numbers varied from 70 to 146, with the mode of 73. Methylcellulose soft agar assay showed that approximately 40% of the HA22T/VGH cells formed colonies. The HA22T/VGH produced tumors in nude mice. Histopathological studies of the tumor revealed the arrangement of hepatoma. Detected by the complement fixation method HA22T/VGH cells secreted ceruloplasmin, Factor B, C3, C4, Gc globulin and alpha 1-acid-glycoprotein. These cells contained the liver associated enzymes: alanine amino transferase, tyrosine amino transferase and gamma-glutamyl transferase. HBsAg and alpha-fetoprotein were not detectable in the HA22T/VGH culture media or cell lysates by the radioimmunoassay. PMID- 6291876 TI - Isolation and identification of swine rotavirus in Taiwan. AB - Large numbers of viral particles resembling rotavirus were detected with negative stained electron microscopy in bacteria free fecal filtrate obtained from 10-day old diarrheal suckling piglets of a conventional pig farm in Taiwan. The clinical signs of vomiting and diarrhea were reproduced in colostrum deprived piglets artificially infected with the fecal filtrates. Rotavirus particles persisted in the fecal samples after two in vivo serial passages, and was not seen in the uninfected control animal. The Cytoplasm of infected jejunal and ileal enterocyte fluoresced when standard anti-porcine rotavirus conjugate was applied in an direct immunofluorescent staining test. In the experimentally infected piglets, moderate villous atrophy of the small intestine was the main microscopic lesion observed. The virus was identified by the above evidence to be rotavirus. PMID- 6291878 TI - Receptors, antibodies and disease. PMID- 6291877 TI - Evolution of human Y-chromosome DNA. AB - We have used human male-specific 3.4 kb Hae III restriction endonuclease fragments to explore the evolutionary history of man's Y-chromosome. We have identified four sets of reiterated sequences on the basis of their relative sequence homology with autosomal DNA. The sequences account for approximately 40% of the human Y-chromosome, are interspersed within the same 3.4 kb Hae III fragments, are heterogeneous and contain all reiterated DNA previously demonstrated to be specific for the Y-chromosome (it-Y DNA). Y-specific 3.4 kb Hae III sequences do not reassociate with either human female or ape DNA at standard reassociation criteria. However, approximately half of it-Y DNA (cross reacting it-Y) reassociates with both human female and ape DNA at reduced reassociation criteria. The remaining half (Y-specific it-Y) retains its specificity for the human Y-chromosome. These two sets of it-Y DNA have distinct reiteration frequencies and thermal stabilities with their Y-chromosome homologs. Non-Y-specific 3.4 kb Hae III sequences reassociate with both human female and ape DNA at standard reassociation criteria. The abundance of these non-Y-specific sequences decreases as a function of their evolutionary distance for man. One subset of non-Y-specific 3.4 kb Hae III sequences forms stable duplexes with human Y-chromosome DNA and with human and ape autosomal DNA. No detectable base mismatch occurs among these homologs suggesting complete conservation of these sequences during primate evolution. The second subset of Non-Y-specific Hae III sequences form stable duplexes with human Y-chromosome DNA but highly mismatched duplexes with human and ape autosomal DNA. The finding that homologs of 3.4 kb Hae III sequences are not found within the Y-chromosome of apes but are only present in autosomes suggests that 3.4 kb Hae III sequences are largely autosomal in origin. Since autosomal homologs of most 3.4 kb Hae III-sequences exhibit a greater degree of divergence that those localized to the Y-chromosome, their evolutionary history seems to be chromosome-dependent. Our findings are not easily correlated with the comparative morphology of primate Y-chromosomes and suggest that sequence rearrangement has been a major event in the evolution of the human Y-chromosome. The significance of the specific interspersion of four sets of reiterated sequences, with distinct evolutionary histories, within a repeating unit specific to the human Y-chromosome is not clear. The apparent conservation of at least some of these reiterated sequences suggests they may be of functional importance. PMID- 6291879 TI - Structure-function relations of the thyrotropin receptor. AB - The thyrotropin (thyroid-stimulating hormone or TSH) receptor is an amphiphilic membrane component with a relative molecular mass of about 200,000 as judged by gel filtration and an isoelectric point close to pH 5. Analyses with chemical, enzymic and affinity probes indicate that the receptor is a glycoprotein containing a disulphide bridge and that the integrity of the disulphide bond is essential for maintaining the structure of the TSH-binding site. Serum from patients with Graves' disease contains antibodies which inhibit the binding of TSH to its receptor and there is considerable evidence that this effect is due to a direct interaction between the antibodies and the receptor. The antibody receptor interaction is probably responsible for the TSH agonist properties of Graves' serum and, similarly, the TSH antagonist properties of the sera from a small number of patients can be explained on the basis of antibody-receptor binding. Although TSH and IgG from Graves' disease patients appear to bind to the same receptor, the relationship between the sites for the two substances is not clearly understood. However, Fab fragments of Graves' IgG are as effective as intact IgG in competing with TSH for the receptor and gel filtration and immunoprecipitation studies indicate that the binding of hormone and antibody to the receptor is mutually exclusive. Current evidence suggests therefore that the binding sites for TSH and TSH receptor antibodies are very closely related and may well be identical. PMID- 6291880 TI - Thyroid antibodies in thyroid diseases. AB - The cell surface expression of the thyroid microsomal antigen is confined to the apical microvilli of the polarized cells in intact or semi-intact thyroid acini. This makes it possible to envisage a pathogenetic role for the commonest antibodies found in thyroid autoimmunity by mechanisms involving complement- and antibody-dependent cytotoxicity. TSH rereceptor antibodies can now be differentiated into those that stimulate hormone synthesis and release (TSI), and those which mimic TSH in its growth-promoting function (TGI). TGI were demonstrated in goitrous Graves' disease and in some patients with euthyroid non toxic goitres, diffuse or nodular, defining a new variety of thyroid autoimmunity. In patients with non-toxic goitre there is hardly any lymphoid thyroiditis and a low incidence of thyroglobulin or microsomal antibodies, the main expression of autoimmunity being the impetus to thyroid hyperplasia and hypertrophy. TSH receptor antibodies of both types, TSI and TGI, can either be stimulating or have blocking effects (TSI block and TGI block). Atrophy of the thyroid gland in myxoedema is due to blocking of the normal pituitary-controlled repair mechanism. In goitrous thyroiditis cell re-growth as a result of increased TSH can occur, as there are no blocking antibodies. In some patients with 'simple' euthyroid goitres, TSI blockers prevent the onset of hyperthyroidism while growth-promoting antibodies give rise to the goitre. PMID- 6291881 TI - Atopy, autonomic function and beta-adrenergic receptor autoantibodies. AB - Atopic individuals (with asthma, allergic rhinitis or atopic eczema) have impaired sensitivity to beta-adrenergic agents. After the finding of antibodies to the beta-adrenergic receptor in the serum of a subject with allergic rhinitis, coded sera from atopic and control subjects were assayed for immunoglobulins that inhibited the specific binding of 125I-labelled hydroxybenzylpindolol to beta receptors in mammalian lung membranes. Antibodies were present in nine of 60 subjects: 3/19 normal control subjects, 1/9 pre-allergic, 4/17 asthma, 0/8 allergic rhinitis, and 1/7 cystic fibrosis patients. Antibodies of the IgG class in these sera were also demonstrated by indirect precipitation of solubilized lung beta-receptors. The autonomic sensitivity of the nine antibody-positive subjects (Ab+) was compared with that of antibody-negative subjects (Ab-). The Ab+ subjects required 15.0 +/- 1.9 ng isoprenaline (isoproterenol) kg-1 min-1 i.v. to increase pulse pressure by at least 22 mmHg (Ab-, 7.7 +/- 0.4; n = 20; P less than 0.001), and 12.4 +/- 1.8 ng isoprenaline kg-1 min-1 i.v. to increase plasma cyclic AMP concentrations by 50% (Ab-, 8.08 +/- 0.62; n = 13; P less than 0.02). Ab+ subjects required 2.06 +/- 0.3% phenylephrine to dilate their pupils (Ab-, 2.55 +/- 0.08; n = 57; P less than 0.05) and 0.61 +/- 0.08% carbachol to constrict their pupils (Ab-, 0.78 +/- 0.03%; n = 57; P less than 0.05). A role for autoantibodies as beta-receptor antagonists was further supported by showing that human lung cells (VA-13 line) cultured in the presence of globulins from Ab+ subjects had a markedly impaired cyclic AMP response to isoprenaline. These results suggest that autoantibodies to beta-receptors play a pathogenetic role in asthma and related disorders. They have important implications for the concept of autoimmunity. PMID- 6291882 TI - Prolactin and growth hormone receptors. AB - The two hormones prolactin and growth hormone exhibit considerable structural homology as well as exerting similar biological effects, especially the primate hormones. One effect of prolactin that deserves greater attention is its action on the immune system including the stimulation of growth of experimental lymphomas, both in vivo and in vitro. One cultured lymphoma cell line has proved to be a very useful model system in which to examine prolactin receptor synthesis and turnover as well as post-receptor mechanisms of action. Prolactin and growth hormone receptors from rabbit mammary gland and liver respectively have been partially purified and characterized. Polyclonal antibodies to prolactin and growth hormone receptors have been generated. The antibodies have been shown to cross-react with prolactin or growth hormone receptors from a number of species, indicating structural homology among receptors as well as hormones. The polyclonal antisera inhibit the action of prolactin in vivo as well as in vitro. In addition, several of the same antisera also mimic the action of prolactin. As yet the presence of autoantibodies to prolactin or growth hormone receptors in human serum samples has not been recognized. PMID- 6291883 TI - Speculations on potential anti-receptor autoimmune diseases. AB - Many autoimmune disorders have a strong tendency to cluster in a single patient or type of patient. Therefore, in those cases in which anti-receptor antibodies are known to be responsible for one of the diseases in the cluster, it is logical to proceed investigatively on the presumption that the aetiology of other members of the cluster may also have an anti-receptor autoantibody basis. This logic is examined by considering examples of clustering in human diseases involving both organ-specific and non-organ-specific autoimmunities. The strong relationship between clustering among autoimmune diseases and the HLA-B8/DRw3 haplotype may provide a marker for anti-receptor autoimmune diseases. PMID- 6291884 TI - Detection of transformed cells using a fluorescent probe: the molecular basis for the differential reaction of fluorescamine with normal and transformed cells. AB - Normal and transformed fibroblasts can be discriminated by a flow cytometry assay on the basis of their differential reaction with fluorescamine. The cause of altered reactivity of transformed cells with this fluorescent probe has been investigated by a detailed analysis of its reaction with chicken embryo fibroblasts transformed by a temperature sensitive mutant of Rous sarcoma virus. The subcellular distribution of fluorescent adducts characterized by cell fractionation and gel electrophoresis procedures supports the hypothesis that transformed cells possess a surface barrier which decreases the accessibility of fluorescamine to reactive macromolecules. The barrier has been identified as being composed at least partly of hyaluronic acid, because of the ability of purified and specific hyaluronidase (from Streptomyces hyalurolyticus) to modulate the response of transformed cells to fluorescamine. Enzyme treatment of transformed cells prior to reaction with fluorescamine causes them to resemble nontransformed cells both in the nature of components labeled and in their fluorescence intensity. It is suggested that fluorescamine monitors an altered surface hyaluronic acid composition which occurs upon transformation. Its significance is discussed in terms of the known physical properties of the molecule and the finding that it is an early event in the process of transformation. PMID- 6291886 TI - [Cloning and the analysis of highly repetitive DNA sequences of the rat liver]. PMID- 6291885 TI - Chronic enterocyte infection with coronavirus. One possible cause of the syndrome of tropical sprue? AB - A man with a gastrojejunostomy and intestinal malabsorption was found to be excreting large numbers of coronavirus-like particles in his stools over a period of at least eight months. Coronavirus-like particles were found in vesicles in degenerating jejunal enterocytes in all of five jejunal biopsies. In a review of electron micrographs, similar structures were found in biopsies from three of 12 patients with classical chronic tropical sprue and in one patient with a sprue like syndrome associated with agammaglobulinaemia. The hypothesis is advanced that infection with this virus may produce enterocyte damage and may be one cause of the syndrome of tropical sprue. PMID- 6291887 TI - [Sarc-specific protein kinase activity of the ribosomal fraction of cells transformed by Rous sarcoma virus]. PMID- 6291888 TI - Cytosine arabinoside stability in intravenous admixtures with sodium bicarbonate and in plastic syringes. AB - The increased use of high-dose cytarabine in the treatment of neoplasms prompted this study on the reconstituted stability of cytarabine in intravenous admixtures with sodium bicarbonate and in plastic syringes. The study of the effect of sodium bicarbonate was prompted by the frequent use of this agent in patients being treated with cytarabine as a means of achieving systemic alkalinization in order to manage hyperuricemia. The stability of reconstituted cytarabine in plastic syringes was measured to determine whether cytosine arabinoside could be reconstituted and stored prior to use. The results of this study indicate that sodium bicarbonate 50 mEq/L in the intravenous solution containing cytarabine has no effect on the chemical stability of cytarabine for at least one week at room temperature or in the refrigerator. Consequently sodium bicarbonate and cytarabine can be coadministered in one intravenous solution. The reconstituted stability of cytarabine at 20 mg/ml and 50 mg/ml is not affected by storage in plastic syringes. PMID- 6291889 TI - [Effects of cimetidine and ranitidine on blood coagulation]. AB - The influence of cimetidine and ranitidine on blood coagulation and plasmatic fibrinolysis were assessed in vitro and in vivo. After an intravenous bolus of 200 mg cimetidine a marked prolongation of plasmatic thrombin time was observed, which however did not occur after 50 mg of ranitidine. In vitro addition of cimetidine (0.12 mg/ml) and ranitidine (0.06 mg/ml) again led to prolongation of plasmatic thrombin time only after cimetidine, reaching double the initial value. This requires application of cimetidine in acute intestinal haemorrhage not as a bolus, but only by drip infusion. Continuous application of 600 mg over 12 hours did not result in changes of coagulation values. In contrast, ranitidine may be administered as a bolus. Bolus injections of 50 to 100 mg did not result in any prolongation of plasmatic thrombin time. The antifibrinolytic effect of both H2 receptor antagonists was investigated in the streptokinase-induced fibrinolysis spindle of the thrombelastogram. Clear-cut antifibrinolytic properties were not demonstrable for either preparation with this method. PMID- 6291890 TI - [Immunity against poliomyelitis in mothers and their newborn infants. Comparison of Hamburg and Bamako (Mali)]. AB - The polio neutralisation test was performed in 199 paired sera from mothers and their newborn infants from the Hamburg area and in 69 paired sera from Bamako (Mali). Antibodies against all 3 poliovirus types were demonstrated in 52.3% of Hamburg and in 78.0% of bamako mothers. The significantly inferior protection of Hamburg mothers was caused by a lower rate of polio 3 antibodies. Only 65.3% of the Hamburg mothers had polio 3 antibodies in contrast to 88.0% of Malian mothers. Newborn infants from Hamburg had significantly higher polio 2 antibody titres than their mothers, newborn infants from Bamako in contrast had significantly lower polio 2 and 3 antibody titres than their mothers. Both groups share an insufficient transferral of polio 3 antibodies. When only titres outside the range of error of the neutralisation test (1 titre step) are considered as different, all 3 polio types in both groups show agreement of titres in mother and child in more than 90%. PMID- 6291891 TI - [The angiotensin-I-converting enzyme. Physiological aspects and clinical significance]. PMID- 6291892 TI - Enucleation blocks the morphological response of glioblasts to glia maturation factor. AB - Culture glioblasts obtained from rat fetuses were enucleated with cytochalasin B. The glia maturation factor stimulated the morphological differentiation of nucleated but not enucleated cells. In contrast, 8-bromo-cyclic AMP stimulated the morphological differentiation of both nucleated and enucleated cells. This distinction implies a difference in the mode of action of the two agents on cultured glioblasts, and suggests that an interaction between the nucleus and cytoplasm is required for expression of the morphological effect of the glia maturation factor. PMID- 6291893 TI - A serotonin agonist-antagonist reversible effect on Na+-K+-ATPase activity in the developing rat brain. AB - In brain of adult and developing rats the Na+-K+-adenosine triphosphatase (Na+-K+ ATPase) system seems to react to serotonin (5-HT) changes induced pharmacologically. A 5-HT agonist (quipazine) elicits a response of the enzyme activity in the cerebral cortex in vivo, which is neutralized with a 5-HT antagonist (methysergide). This effect was observed from day 21 to adulthood. Also in a state of 5-HT receptor hypersensitivity (rats treated early with 5,6 dihydroxytryptamine), the response of Na+-K+-ATPase to the 5-HT agonist was higher than without neurotoxic lesion of 5-HT paths. These data suggest an involvement of the Na+-K+-ATPase system in 5-HT receptor sensitivity in the rat brain. PMID- 6291894 TI - Chronic mucocutaneous candidiasis treated with ketoconazole. PMID- 6291895 TI - The importance of aftercare for the success or unsuccess in periodontal therapy - a conceivable model. PMID- 6291896 TI - [Realization of aftercare in practice]. PMID- 6291897 TI - [Electron microscopic study of nuclear inclusions in the cells of experimental malignant lung tumors]. PMID- 6291898 TI - Sequestration of an early-release pool of growth hormone and prolactin in GH3 rat pituitary tumor. PMID- 6291899 TI - Resumption of pulsatile luteinizing hormone release after alpha-adrenergic stimulation in aging constant estrous rat. PMID- 6291900 TI - Metabolism of 25-hydroxycholesterol by rat luteal mitochondria and dispersed cells. AB - The metabolism of 25-hydroxycholesterol (25-OH-cholesterol) to progestins by mitochondria and dispersed cells prepared from ovaries of PMSG-hCG-primed rats was studied. Mitochondria converted [3H]25-OH-cholesterol into [3H]pregnenolone and [3H]progesterone. Unlabeled 25-OH-cholesterol also stimulated mitochondrial steroidogenesis in a dose-dependent, saturable fashion. A direct relationship between rates of steroid synthesis in the presence of 25-OH-cholesterol and mitochondrial cytochrome P-450 levels was found. Although steroid production and cytochrome P-450 content per milligram protein were higher in mitochrondia prepared from ovaries removed on day 8 post hCG than on either day 1 or day 14, steroid production per nanomole cytochrome P-450 was similar. Treatment of rats with hCG 1 h before killing significantly increased mitrochondrial steroid synthesis from endogenous substrate but had no effect on metabolism of 25-OH cholesterol. Dispersed cells increased progestin production by 6-fold when incubated with 25-OH-cholesterol. The effects of 25-OH-cholesterol were dose dependent and saturable. While both LH and (Bu)2cAMP stimulated progestin synthesis from endogenous substrate, secretion of progestins with these agents reached levels only 60% of those observed in the presence of 25-OH-cholesterol. Neither LH nor (Bu)2cAMP altered the metabolism of the dydroxysterol by the cells nor did cycloheximide, which substantially inhibited progestin secretion in the absence of the hydroxysterol. However, animoglutethimide did block the stimulation of steroidogenesis by 25-OH-cholesterol. We conclude that 25-OH cholesterol is an effective steroidogenic substrate for rat luteal tissue. With its use, information regarding the maximal capacity of luteal tissue to produce progestins in vitro can be obtained. PMID- 6291901 TI - Thyroid hormone administration in vivo regulates the activity of hepatic glycogen phosphorylase phosphatase. AB - Short term (48 h) administration in vivo of either T4 or T4, but not the biologically inactive D-isomer of T3, was associated with a decrease in basal glycogen phosphorylase alpha activity and an increase in phosphorylase alpha phosphatase activity of rat hepatocytes. This influence of thyroid hormones on hepatic phosphorylase alpha and phosphorylase phosphatase activities was shown to be dose dependent. As little as 0.0025 mg T3/kg BW administered in vivo at 48, 24, and 3 h before liver excision increased the phosphatase activity by 20%. The administration of 0.25 mg T3/kg BW on the same schedule increased the phosphatase activity by nearly 100%. PMID- 6291902 TI - Catecholamine-stimulated potassium transport in erythrocytes from normal and hyperthyroid turkeys: quantitative relation between beta-adrenergic receptor occupancy and physiological responsiveness. PMID- 6291903 TI - Interaction of adenosine 3',5'-monophosphate derivatives with the gonadotropin releasing hormone receptor on pituitary and ovary. PMID- 6291904 TI - Increased secretion of vasopressin and adenosine 3',5'-monophosphate from hypothalamic-posterior pituitary units of spontaneously hypertensive rats. AB - To determine if the increased vasopressin (AVP) levels observed in the blood and urine of spontaneously hypertensive rats (SHR) are a response to peripheral factors that may influence AVP release or are a consequence of altered hypothalamic-neurohypophysial activity, AVP release from hypothalamic neurohypophysial units was studied using an in vitro perifusion system. Spontaneous and 30 mM K+-stimulated AVP release was significantly greater from tissue of SHR rats than from those of WKYN. Adenosine (10(-5) M) added to the perifusion medium increased AVP release into the perifusate in both strains, even through AVP release into the perifusate was greater in tissue of SHR rats. Measurement of AVP content revealed that hypothalamic AVP was lower in SHR rats, whereas the neural lobes of the SHR contained a significantly higher concentration of AVP compared to the tissue of WKYN rats. In addition, exposing tissue from SHR rats to 30 mM K+ stimulated an increase in cAMP release into the perifusate, whereas tissue from WKYN rats did not increase cAMP release above basal level. These data suggest that central nervous system-mediated hyperresponsiveness is the basis for the increased AVP secretion that occurs in the SHR rat and are consistent with reports of a hypersensitive hypothalamic anterior pituitary axis in these animals. PMID- 6291905 TI - gamma-Aminobutyric acid receptors in anterior pituitary and brain areas after median eminence lesions. AB - Previous results have indicated that the median eminence (ME) plays a key role in the regulation of estrogen and dopamine receptors in the anterior pituitary (AP). Since 3H-labeled gamma-aminobutyric acid ([3H]GABA) receptors have been described in the pituitary as well as in the brain, the aim of the present work was to investigate whether those receptors are also under hypothalamic control. Ovariectomized adults rats were divided into two groups. In the first one, the ME was lesioned by an anodic current (MEL); the second group consisted of sham operated controls. Animals were used 7-14 days thereafter to study [3H]GABA binding in a crude mitochondrial membrane fraction prepared from the anterior pituitary, the hypothalamic-preoptic suprachiasmatic area, or the frontoparietal cortex. Binding of [3H]GABA was a saturable process, with high affinity in all studied structures. In AP Scatchard analysis of the saturation curves indicated similar Kd values in sham and MEL rats; the maximal number of binding sites increased from 0.65 +/- 0.005 pmol/mg protein in controls to 1.05 +/- 0.006 pmol/mg protein in MEL rats (P less than 0.001). In the hypothalamus, Kd values were similar in both groups, but the maximum number of binding sites (Bmax) increased about 50% in MEL animals. In the frontoparietal cortex, Kd values were similar in control and lesioned animals; but the Bmax decreased by 40% after the lesion. Animals with lesions showed hyperprolactinemia and a reduction in serum levels of LH, FHS, and TSH. Thus, Kd values were of similar magnitude in all studied regions and were not altered by ME destruction. Bmax values in controls were higher in the cerebral cortex than in the hypothalamus and AP. After ME lesions, there was an increment of [3H]GABA binding in AP and hypothalamus and a reduction in cerebral cortex. The data indicate that the ME participates in the regulation of [3H]GABA binding in AP and also suggest that it plays a role in regulating GABA receptors in various regions of the brain. PMID- 6291906 TI - Prolactin inhibition of luteinizing hormone-stimulated androgen synthesis in ovarian interstitial cells cultured in defined medium: mechanism of action. AB - The mechanism by which PRL acts on ovarian interstitial cells to inhibit androgen synthesis was examined using primary cultures of ovarian cells from hypophysectomized immature rats grown in serum-free medium. In the presence of LH, the cultured interstitial cells showed a 200-fold increase in androgen production, of which androsterone was the principal metabolite. The addition of highly purified PRL (100 ng/ml) markedly inhibited (98%) the LH-stimulated androsterone accumulation. The ED50 of PRL action was calculated to be 1.3 +/- 0.4 ng/ml. The inhibition of androsterone production by PRL was rapid (t 1/2 = 75 min), not readily reversible, and could be evoked at any time during the culture period. The effects of PRL on LH-stimulated androgen production were not due to changes in [125I]iodo-hCG binding. LH-stimulated adenylate cyclase, cell number, or cell viability. As with LH, prostaglandin E2, cholera toxin, or 8-bromo cAMP also caused marked increases in androsterone synthesis, and these effects were blocked (98-99%) by PRL. Studies on the metabolism of steroid hormones revealed that PRL decreased LH-stimulated androsterone and 5 alpha-androstane-3 alpha, 17 beta-diol accumulation by 99%, androstenedione by 94%, testosterone by 90%, dehydroepiandrosterone by more than 80%, pregnenolone and 17 alpha hydroxyprogesterone by 80%, 17 alpha-hydroxypregnenolone by 84%, and progesterone by 71%. Binding experiments demonstrated the presence of a single class of high affinity (Kd = 2.42 x 10(10) M), low capacity (2.37 fmol/10(6) cells) [125I]iodo PRL-binding sites in the interstitial cells, suggesting that such receptors mediate the inhibitory action of PRL. It is inferred from these results that PRL antagonizes the stimulatory effects of LH on ovarian androgen biosynthesis by inhibiting a step distal to cAMP formation and before or at the cholesterol side chain cleavage step. PMID- 6291907 TI - Metabolism of vitamin D3 in nephrectomized pigs given pharmacological amounts of vitamin D3. AB - The metabolism of vitamin D3 was studied in bilaterally nephrectomized pigs and cotnrol pigs maintained for 8-10 days by daily peritoneal dialysis. Two or three pharmacological doses of vitamin D3 were given im, one on day 0 and the others on days, 4, 6, or 7 of the 8- to 10-day experimental periods. Nephrectomized pigs had extremely high plasma concentrations of 24,25-dihydroxyvitamin D3 [24,25 (OH)2D3] and 25,26-(OH)2D3 at the end of the 8- to 10-day periods; similar levels were present in control pigs. However, nephrectomized pigs had no detectable increase in plasma 25-hydroxyvitamin D3 (25OHD3)-26,23-lactone concentrations throughout the experiment, in contrast with the high plasma concentrations of 25OHD3-26,23-lactone in the control pigs. Much higher plasma 25OHD3 concentrations were present in the nephrectomized pigs than in the control pigs. Plasma 1,25-(OH)2D concentrations in nephrectomized and control pigs were similar during most of the experiment, except the anephric pigs usually had significantly higher plasma 1,25-(OH)2D concentrations for several days after the vitamin D3 injections. These studies demonstrate extrarenal production of 24,25-(OH)2D3, 25,26-(OH)2D3, and possibly 1,25-(OH)2D, and kidney-dependent synthesis of 25OHD3 26,23-lactone in the pigs. PMID- 6291908 TI - Effects of hormones on mammary adenosine 3',5'-monophosphate levels and metabolism in normal and adrenalectomized lactating rats. AB - Prostaglandin E1 (PGE1) T4, and epinephrine (Epi) were incubated with mammary tissue slices from normal and adrenalectomized (Adx) rats and Adx rats receiving cortisol (Adx + C). PGE1, T4, and the combination of Epi, T4, and PGE1 increased mammary slice cAMP levels in all groups. No differences were observed among treatment groups in basal cAMP levels or responses to hormones. Glucose-1-14C oxidation and incorporation into lipids were stimulated by PGE1, T4, and the combination of EPi, T4, and PGE1 in normal rats. Responses were depressed by adrenalectomy and reversed by cortisol. PGE1, T4, and the combination of EPi, T4, and PGE1 depressed glucose-6-14C oxidation. Fatty acid synthesis from glucose-6 14C was stimulated by Epi, PGE1, and T4 in tissues from normal and Adx + C rats, but not in those from Adx rats. Glycerol synthesis was unchanged. Pyruvate-1-14C and pyruvate-2-14C oxidation were uneffected by adrenalectomy or hormones. Fatty acid synthesis was depressed by adrenalectomy and reversed by cortisol. T4 and the combination of Epi, T4, and PGE1 depressed triacylglycerol and fatty acid synthesis in all groups. Glycerol synthesis was unchanged. Hormonal effects were much more pronounced when glucose was the substrate. This suggests that cAMP effects on glucose metabolism are manifest at a site between hexose-P and pyruvate, and further, that a primary effect of adrenalectomy may be to reduce the responsiveness of this site to cAMP. Linear regression analyses comparing cAMP levels with glucose and pyruvate metabolic parameters further support this conclusion. PMID- 6291909 TI - Development of antral follicles in cattle after prostaglandin-induced luteolysis: changes in serum hormones, steroids in follicular fluid, and gonadotropin receptors. AB - Changes in growth, concentrations of steroids in follicular fluid, and numbers of gonadotropin receptors in follicles 6 mm or more in diameter were examined 12, 24, 36, 48, 60, and 72 h after prostaglandin (PG)-induced luteolysis in heifers. Follicles with higher concentrations of estradiol than progesterone and androgens in follicular fluid (FF) were classified as estrogen active (E-A). Follicles with higher concentrations of progesterone or androgens than estradiol in follicular fluid were classified as estrogen inactive (E-I). Although E-A and E-I follicles were similar in size 12-72 h after PG treatment, before the preovulatory surge of LH, numbers of granulosa cells, estradiol concentrations, and capacities of follicles to specifically bind gonadotropins were markedly lower in E-I follicles. Most E-I follicles had histological signs of atresia, whereas E-A follicles did not. Sizes, concentrations of steroids in FF, and specific binding of gonadotropins to follicle cells did not change in E-A or E-I follicles 12-72 h after PG treatment but before the preovulatory surge of LH. However, diameters of E-A follicles were highly correlated with concentrations of estradiol and progesterone in FF and with capacities of granulosa and thecal cells to specifically bind [125I] iodo-hCG. In contrast, diameters of E-A follicles were highly negatively correlated with concentrations of testosterone in FF and with capacities of granulosa cells to specifically bind [125I] iodobovine FSH. We conclude that E-I follicles are atretic and E-A follicles are potentially ovulatory follicles. In addition, after a single injection of PG during an estrous cycle in heifers, the growth the differentiation of ovulatory follicles were not synchronized. PMID- 6291910 TI - Adrenocorticotropin, beta-endorphin, and beta-lipotropin in normal thyroid and lung: possible implications for ectopic hormone secretion. AB - The expression of the proopiomelanocortin (POMC) gene by normal lung and thyroid was examined by measurement of the content of ACTH, beta-lipotropin (beta LPH), and beta-endorphin (beta EP) in porcine lung and thyroid tissue. Acid extracts of normal porcine lung and thyroid tissue each contained appreciable amounts of immunoreactive (ir) ACTH, ir-beta LPH, and ir-beta EP. The content of ir-beta LPH in both tissues exceeded by severalfold, on a molar basis, the content of ir-ACTH and ir-beta EP, suggesting that the common precursor POMC was processed predominantly to peptides other than ir-ACTH and ir-beta EP. A porcine thyroid extract (Calcitare, porcine calcitonin, Armour) showed equivalent levels of beta EP-like immunoreactivity and bioactivity, measured by opiate radioreceptor assay; in contrast, ACTH-like bioactivity, measured by rat zona fasciculata steroidogenesis, was only 4% of ACTH-like immunoreactivity. On reversed phase high performance liquid chromatography, Calcitare showed multiple peaks of ACTH like immunoreactivity, one of which coeluted with porcine ACTH-(1-39), and two much smaller peaks of beta EP-like immunoreactivity, of which the smaller coeluted with porcine beta EP. These data suggest that both lung and thyroid gland synthesize POMC, which in normal tissue is usually predominantly processed to species other than ACTH and beta EP. Ectopic secretion of ACTH and beta EP by lung and thyroid neoplasms may thus represent the loss of a system(s) normally responsible for processing the precursor beyond ACTH and beta EP. PMID- 6291911 TI - A novel method for localization of gonadotropin releasing hormone receptors. AB - Two 125I-labeled analogs of GnRH, [acidobenzoyl-D-Lys6]GnRH (I) and [D-Lys6]GnRH (II) were used for the localization of GnRH receptors in pituitary cells. The analogs exhibited high binding affinity to pituitary membrane preparations and after photoactivation (analog I) or cross-linking with glutaraldehyde (analog II), these analogs are bound covalently to pituitary cells. The distribution of the labeled hormones by light and electron microscopic autoradiography indicated that after exposure of pituitary cells to 125I-labeled hormones at 4 C (90 min), most of the labeled hormones were associated with the cell surface membrane, while at 37 degrees C (30 min) most of the cell-bound labeled hormones were internalized. PMID- 6291912 TI - Subcellular localization of the receptor for gonadotropin-releasing hormone in pituitary and ovarian tissue. AB - The subcellular localization of GnRH receptors in weanling rat anterior pituitary and ovarian tissue was determined by radioligand binding in biochemically defined fractions prepared by differential and density centrifugation. Morphological identification of membrane organelles or fragments, visualized by electron microscopy, confirmed the biochemical characterization. The greatest amount of specific ligand binding in both tissues was measured in the crude membrane fraction, while small amounts were detected in other fractions. The distribution of pituitary binding sites in sucrose gradient subfractionation of crude membranes correlated with the distribution of plasma membrane (assessed by 5' nucleotidase activity and electron microscopy), but not that of secretory granules (identified by immunoreactive LH) or lysosomes (acid phosphatase activity). These data suggest that the GnRH-binding sites detected by radioligand binding assay in pituitaries of rats not previously exposed to GnRH are localized almost exclusively in the plasma membrane fraction. PMID- 6291913 TI - Comparison of effects of adrenocorticotropin and Lys-gamma 3-melanocyte stimulating hormone on steroidogenesis, adenosine 3',5'-monophosphate production, and phospholipid metabolism in rat adrenal fasciculata-reticularis cells in vitro. AB - Synthetic bovine Lys-gamma 3MSH was found to potentiate the steroidogenic action of ACTH during incubation of rat adrenal fasciculata-reticularis cells in vitro. On the other hand, Lys-gamma 3MSH did not increase basal levels of or ACTH induced (submaximal) increases in cellular concentrations of cAMP or phosphatidylinositol. Thus, Lys-gamma 3MSH does not appear to simply increase the overall action of ACTH, and moreover, it appears to potentiate steroidogenesis by a mechanism that is considerably different from that employed by ACTH. Observance of an effect of ACTH and failure to observe an effect of gamma 3MSH on adrenal phosphatidylinositol are in keeping with our previous postulation that phospholipids in the phosphatidate-inositide cycle play an important role in promoting cholesterol side-chain cleavage, since ACTH, but not gamma 3MSH, reportedly increases cholesterol side-chain cleavage. In addition, we also presently observed that Lys-gamma 3MSH can markedly increase corticosterone production in the face of a fixed, relatively small, submaximal, ACTH-induced increase in phosphatidylinositol. Thus, if gamma 3MSH enhances steroidogenesis by increasing free cholesterol availability, the latter may require another factor to initiate steroidogenesis, but is, nevertheless, an important determinant of the rate of steroidogenesis. PMID- 6291914 TI - Development of nonovulatory antral follicles in heifers: changes in steroids in follicular fluid and receptors for gonadotropins. AB - The objective of this study was to examine changes in steroid concentrations in follicular fluid (FF) and in gonadotropin receptors in nonovulatory follicles that develop after ovulation in heifers. Six groups of six to seven heifers were ovariectomized on days 3, 5, 7, 9, 11, and 13 of an estrous cycle. Follicles 6 mm or more in diameter were separated into two classes, estrogen-active (E-A) and estrogen-inactive (E-I). E-A follicles had higher concentrations of estradiol than progesterone and androgens in FF and a low incidence of atresia. E-I follicles had higher concentrations of progesterone or androgens than estradiol in FF and a high incidence of atresia. Diameters, numbers of granulosa cells, and capacities of granulosa cells and thecal tissue to bind [125I]iodo-hCG increased in E-A follicles from days 3-7. During this time, diameters, numbers of granulosa cells, concentrations of estradiol, and capacities of granulosa cells and thecal tissue to bind gonadotropins were greater in E-A than E-I follicles. On days 9, 11, and 13, each heifer had only a single E-I follicle. Compared with E-A follicles on day 7, E-I follicles on days 9, 11, and 13 were smaller and had less granulosa cells, less estradiol in FF, and a lower capacity to bind [125I]iodo hCG. After day 7, E-A follicles were not observed until day 13. We conclude that at least two periods of growth and atresia of follicles occur between days 3-13 of an estrous cycle in heifers. One is during days 3-7 when a single E-A follicle 6 mm or more in diameter develops and all other E-I follicles regress. The other is between days 7 and 13 when the E-A follicle from day 7 becomes an E-I follicle and regresses, and another E-A follicle develops. PMID- 6291915 TI - Plasma renin activity during infusion of epinephrine into the celiac and superior mesenteric arteries in dogs. AB - Previous experiments have demonstrated that increments of circulating epinephrine concentration within the physiological range elevate PRA and that this effect apparently is mediated by extrarenal beta-adrenoceptors. The present experiments were designed to test the possibility that the receptors mediating the PRA response to epinephrine are located in the splanchnic region. Accordingly, adult dogs were anesthetized, and catheters were placed for recording blood pressure, withdrawal of blood samples, and collection of urine. Infusion catheters were placed in a femoral vein and in the celiac and superior mesenteric arteries. After a control period, epinephrine was infused for 45 min at a total rate of 25 ng kg-1 min-1 either iv or directly into the celiac and superior mesenteric arteries (12.5 ng kg-1 min-1 into each artery simultaneously). Renal perfusion pressure was kept at the control level during the infusion period by means of an adjustable suprarenal aortic clamp. PRA rose from a mean control plus recovery value of 4.5 +/- 0.9 ng ml-1 h-1 to 8.7 +/- 0.7 ng ml-1 h-1 during iv epinephrine infusion. In contrast, PRA remained unchanged (3.9 +/- 0.8 vs. 4.3 +/- 1.1 ng ml 1 h-1) in response to infusion of epinephrine directly into the celiac and superior mesenteric arteries. The data indicate that the receptors mediating epinephrine-induced increases in PRA are not located in organs perfused by the celiac and superior mesenteric arteries. PMID- 6291916 TI - The ovary: a target organ for 1,25-dihydroxyvitamin D3. AB - Using both clonal Chinese hamster ovary (CHO) cells in culture and the hen ovary, we have searched for the presence of specific 1,25-dihydroxyvitamin D3 [1,25 (OH)2D3] receptors. Receptor analyses were carried out on high salt cytosols of CHO cells and high salt extracts of hen ovarian nuclei that were originally isolated in low salt buffer. Both CHO and hen ovary contain a specific high affinity 1,25-(OH)2D3 receptor (Kd = 10(-10) - 10(-11) M), which sediments (3.3S) in high salt sucrose gradients identically to the chick intestinal receptor for 1,25-(OH)2D3. This 3.3S macromolecule from both sources absorbed to DNA-cellulose at 0.1 M KCl and eluted during a linear salt gradient at 0.2-0.22 M KCl, a property characteristic of the 1,25-(OH)2D3 receptor. Saturation analysis indicated that there are approximately 2000 copies of the receptor molecule per CHO cell. We also investigated the effect of 1,25-(OH)2D3 on ovarian cell growth in monolayer culture of CHO cells. Significant inhibition of CHO cell growth (up to 60%) was observed in the presence of physiological (100 pM) levels of 1,25 (OH)2D3 in the culture medium. This inhibition of growth was dose dependent and was accompanied by a parallel decrease in total cell protein. Concentrations of 25-hydroxyvitamin D3 and 24,25-dihydroxyvitamin D3 as high as 1 nM did not affect CHO cell growth, indicating that, like receptor binding, cell proliferation is selectively influenced by 1,25-(OH)2D3 over other vitamin D metabolites. These data demonstrate that ovarian cells in mammals and birds possess the 1,25-(OH)2D3 receptor which may play a role in the effect of 1,25-(OH)2D3 on the growth of these cells in culture. PMID- 6291917 TI - Increases in brain nuclear triiodothyronine receptors associated with increased triiodothyronine in hyperthyroid and hypothyroid rats. PMID- 6291919 TI - Mice are insensitive to the antitesticular effects of luteinizing hormone releasing hormone agonists. AB - Treatment of male rats with [(imBzl)-D-His6, Pro9-NEt]LHRH or [D-Trp6,Pro9 NEt]LHRH, potent agonists of LHRH, led to a marked decrease in serum testosterone levels and a reduction in testicular LH receptor concentration. Similar treatment of mice showed that they were resistant to the antitesticular effects of the LHRH agonists. To further explore the differences between rats and mice, the direct antitesticular effects of these peptides were investigated in hypophysectomized animals. Hypophysectomized rats and mice were given ovine FSH (50 micrograms), with or without a LHRH agonist (10 micrograms), daily for 5 days. On day 6, the testicular steroidogenic response to hCG was studied. In these studies the in vivo as well as the in vitro steroidogenic response of rat testes to hCG were inhibited by the LHRH analogs. In contrast, pretreatment of mice with the LHRH analogs did not affect their testicular steroidogenic response. Binding studies with the [125I]LHRH analog demonstrated receptors for this peptide on Leydig cells from adult rats. Receptors for LHRH were not, however, detectable on murine Leydig cells. These results suggest that one of the reasons for the lack of an antitesticular effect of LHRH agonists in mice may be due to the inability of these peptides to have a direct effect on testes and may relate to a lack of LHRH receptors. PMID- 6291918 TI - Studies on corticotropin-induced desensitization of normal rat adrenocortical cells. AB - The effects of prior exposure of normal rat adrenocortical cells to ACTH on the responsiveness of the cells to subsequent stimulation with the hormone have been studied. ACTH induces a time- and concentration-dependent refractoriness of both cAMP formation and steroidogenesis. Desensitization of either response was observed only upon activation of the response. Thus, both ACTH and 8-Br-cAMP caused desensitization of the steroidogenic response. The ACTH-induced desensitization of steroidogenesis, however, was completely prevented by blocking the steroidogenic action of ACTH with aminoglutethimide during exposure of cells to the hormone. Aminoglutethimide had no effect on ACTH-induced desensitization of the cAMP response. Studies with analogs of the hormone also confirmed that induction of desensitization of the steroidogenic response is independent of the desensitization of the cAMP response. Binding studies showed that the insignificant decrease in ACTH receptors could not account for the large changes in the responsiveness induced by prior exposure of cells to ACTH. Desensitization of the steroidogenic response appears to result from a defect in the rate limiting first step of the steroidogenic pathway, namely conversion of cholesterol to pregnenolone. PMID- 6291920 TI - 3,5,3'-triiodothyronine nuclear receptor in chick embryo. Properties and ontogeny of brain and lung receptors. PMID- 6291921 TI - Opioids modulate pituitary receptors for gonadotropin-releasing hormone. AB - The number of pituitary GnRH receptors (GnRH-BC) is stable throughout the day in ovariectomized-estradiol treated rats, but undergo an acute transient reduction prior to the afternoon gonadotropin surge. To investigate the mechanisms controlling GnRH-BC we studied the effects of opioid-active compounds in this model. Morphine, given at 1400h, abolished both the LH surge and the preceding fall in GnRH-BC. Morphine given at 0900h increased GnRH-BC 30 min later, and this effect was abolished by simultaneous administration of naloxone. Naloxone alone produced an acute transient fall in GnRH-BC of similar magnitude to that seen before the spontaneous LH surge. These data suggest that alterations in endogenous opioid activity can modulate GnRH receptors and may form part of the mechanisms which initiate the afternoon gonadotropin surge. PMID- 6291922 TI - Opiatergic control of LH secretion is eliminated by gonadectomy. AB - We have studied the stimulatory effects of naloxone and the inhibitory influence of the opiate peptide, FK 33-824, on LH secretion in the gonadectomized rat. Our results indicate that endogenous and exogenous opiate involvement in LH release disappear coincident with the removal of gonadal steroid feedback. At 7 days post surgery naloxone is no longer able to stimulate LH secretion in male or female rats. Similarly, by 7 days in the male, and 21 days in the female, FK 33-824 is unable to inhibit LH secretion. We conclude that the coupling of hypothalamic opiate receptors to the LH regulatory mechanisms is dependent upon gonadal steroids. The most important of these appear to be estradiol and testosterone, since careful priming of long-term gonadectomized rats with these steroids is able to largely restore the LH responses to naloxone and FK 33-824. PMID- 6291923 TI - In vitro studies on basal and stimulated prolactin release by rat anterior pituitary: a possible role for calmodulin. AB - The mechanism by which PRL is released from mammotrophs is a calcium-dependent process. Although calcium seems to function as a second messenger, its regulatory mechanism in PRL release has not been clarified. The binding of calcium to calmodulin and the activation of calmodulin-dependent enzymes have been suggested to be important steps during stimulus-secretion coupling in various cells. In the present work we investigated the in vitro effect of penfluridol, a potent neuroleptic that also possesses the ability to inhibit calmodulin's biological activity, on basal and stimulated PRL release. The effect of pimozide and haloperidol on basal PRL release was also investigated. Penfluridol, pimozide, and haloperidol inhibited basal PRL secretion in a dose-related manner, with the EC50 ranging from 0.5-1 microM for penfluridol to 1-2 microM for pimozide and more than 3 microM for haloperidol. These concentrations are similar to those necessary for the inactivation of calmodulin-dependent enzymes in vitro. Ionophore A-23187, a compound whose ability to mobilize extracellular calcium is not affected by neuroleptics, stimulated PRL secretion in vitro. This effect, however, was blocked by penfluridol pretreatment. The site of action of penfluridol may occur after calcium mobilization, with calmodulin a possible target for penfluridol's inhibitory action on PRL secretion. TRH, K+, (Bu)2cAMP, and theophylline, compounds that affect calcium mobilization, also significantly stimulated PRL release. The coincubation of varying concentrations of penfluridol with 70 nM TRH, 50 mM K+, 3 mM (Bu)2cAMP, or 5 mM theophylline resulted in a dose related inhibition of secretagogue-stimulated PRL secretion. Perifusion of dispersed anterior pituitary cells with 1 microM penfluridol reduced the ability of 70 nM TRH to stimulate PRL release by approximately 50%, whereas removal of the penfluridol perifusion allowed the cells to again be fully responsive to TRH. These results are consistent with the hypothesis that calmodulin is involved in the stimulus-secretion coupling of PRL. PMID- 6291925 TI - A rare complication after Haring-tube implantation. PMID- 6291924 TI - The relationship of growth and adenylate cyclase activity in cultured thyroid cells: separate bioeffects of thyrotropin. PMID- 6291926 TI - Experimentally induced mesothelioma in white rats in response to intraperitoneal administration of amorphous crocidolite asbestos: preliminary report. PMID- 6291927 TI - The pathological effects of prolonged asbestos ingestion in rats. PMID- 6291928 TI - Pleural mesotheliomas in Sprague-Dawley rats by erionite: first experimental evidence. PMID- 6291929 TI - Cutaneous effects of exposure to polybrominated biphenyls (PBBs): the Michigan PBB incident. PMID- 6291930 TI - On the relationship between rate of ATP synthesis and H+ electrochemical gradient in rat-liver mitochondria. AB - The relationship between rate of ATP synthesis, JATP, and value of the proton electrochemical gradient, delta mu H, has been analyzed in intact mitochondria. Onset of phosphorylation causes a depression of delta mu H of 1.5 kJ/mol. There is a close parallelism between inhibition of JATP and restoration of delta mu H to its state-4 value during titrations with oligomycin or atractyloside. Titrations with ionophores display the following features: (a) delta mu H can be depressed by 3-4 kJ/mol by valinomycin + K+ without affecting the rate of ATP synthesis; (b) uncouplers abolish JATP completely while depressing delta mu H by 3 kJ/mol; (c) complete abolition of ATP synthesis by inhibitors of electron transport is accompanied by a depression of delta mu H of only 1 kJ/mol. The results indicate that: (a) there is a close functional relationship between redox and ATPase H+ pumps, whereby inhibition of electron transfer is accompanied by simultaneous inhibition of the ATPase H+ pumps; and (b) uncoupling of oxidative phosphorylation is not due to depression of delta mu H per se. The consistence of the present data with either a chemiosmotic model where delta mu H is the sole and obligatory intermediate for energy coupling, or models where there is a direct transfer of energy between the two pumps is discussed. PMID- 6291931 TI - The structural isomerisation of human-muscle adenylate kinase as studied by 1H nuclear magnetic resonance. AB - Human muscle adenylate kinase (ATP:AMP phosphotransferase, EC 2.7.4.3.) was studied by 1H-nuclear magnetic resonance spectroscopy. The C-2 and C-4 proton resonances of the active-center histidine His-36 could be identified; the pK of His-36 was determined as 6.1. The pK of His-189 is very low (4.9) although it is located at the surface of the protein. Other resonance lines are discussed in comparison with NMR spectra of porcine adenylate kinase [McDonald et al. (1975) J. Biol. Chem. 250, 6947-6954]. A pH-dependent structural isomerization as shown by X-ray crystallography in the pig enzyme [Pai et al. (1977) J. Mol. Biol. 114, 37-45] was not observed for human adenylate kinase in solution. However, the binding of adenosine(5')pentaphospho(5')adenosine (Ap5A), a bisubstrate inhibitor, to adenylate kinase causes an overall change of the NMR spectrum indicative of a large conformational change of the enzyme. The exchange rate (koff) for Ap5A was estimated as 10 s-1 and decreases by addition of Mg2+. On the basis of these values and the known dissociation constant it is likely that the binding of Ap5A is a diffusion-controlled process kon being 10(8) M-1 s-1. In conclusion, the system Ap5A/Mg2+/human adenylate kinase, which has been studied by NMR spectroscopy and X-ray diffraction in parallel, is suitable for analyzing the induced fit postulated by Jencks for all kinase-catalyzed reactions. PMID- 6291932 TI - An affinity label for alpha 2-adrenergic receptors in rat brain. AB - Clonidine, a potent and highly selective alpha 2-adrenergic agonist of the central nervous system, was modified. Insertion of the strong alkylating isothiocyanate group (NCS) group, at its aromatic residue, makes clonidine a potential affinity label of the alpha 2-adrenergic receptors. In displacement of [3H]clonidine and p-[3H]aminoclonidine from rat brain membrane preparations, clonidine-NCS demonstrates high affinity for the alpha 2-adrenergic receptors (Kd = 50 mM). The covalent labelling of the central alpha 2-receptors requires higher concentrations of the irreversible ligand (1-70 microM), thus indicating possible non-productive interactions at the environment of the receptor site. Only partial protection of the receptors is observed with a reversible alpha 2-agonist. The new clonidine analog appears to be a general ligand for the alpha 2-adrenergic receptors and might serve as a potential affinity probe for these receptors. PMID- 6291933 TI - Effect of caffeine on glucose-induced inactivation of gluconeogenetic enzymes in Saccharomyces cerevisiae. A possible role of cyclic AMP. AB - The mechanism of catabolite inactivation of three gluconeogenetic enzymes, fructose-1,6-bisphosphatase, cytoplasmic malate dehydrogenase and phosphoenolpyruvate carboxykinase, has been studied in the yeast Saccharomyces cerevisiae. The glucose-induced inactivation of the three enzymes is remarkably retarded by preincubation of the cells with different caffeine concentrations; however, a full conservation of activity has never been obtained, even at the highest drug concentration. Caffeine modifies the metabolic effects produced in the yeast cell by exposure to glucose. It reduces the consumption rate of glucose; changes the glycolytic intermediate pattern, giving rise to a crossover point at the level of the phosphofructokinase/fructose-bisphosphatase cycle; and increases the ATP level and the energy charge. Moreover, it substantially reduces the peak of intracellular cAMP content that immediately follows glucose entry; the magnitude of this effect is dependent on the drug concentration. The effect on the change of intracellular cAMP level appears, among all metabolic effects determined by caffeine, the only plausible one to explain the interference with catabolite inactivation of enzymes. Actually a strong negative correlation between residual activity of each of the three investigated enzymes and intracellular cAMP level has been demonstrated. The existence of a common mechanism of action of cAMP, as the mediating factor for catabolite inactivation of all three enzymes, is proposed. PMID- 6291935 TI - Activity of cyclic-AMP phosphodiesterase in permeabilised cells of Bakers' yeast. AB - Yeast cyclic-AMP phosphodiesterases were assayed in situ, using cells permeabilised with cytochrome c, to get information about the kinetics of these enzymes at the high concentrations of macromolecules occurring in vivo. Protamine treatment was not suitable, because it perturbed the intracellular localisations of both the (Mg-dependent) low-Km enzyme and the (EDTA-insensitive) high-Km enzyme. The pH-dependence of Km and V for EDTA-insensitive activity in situ agreed well with the behaviour of pure high-Km enzyme, except that near pH 8 Hofstee plots were bent slightly upwards both for activity in situ and with crude broken-cell preparations. Hofstee plots of Mg-dependent activity in situ were distinctly concave, and could be resolved mathematically into two activities, one (accounting for about 30% of the Mg-dependent V) with a Km close to the value in vitro of 0.2 microM, and the other with an apparent Km of 3 microM. The 3 microM Km activity probably represents a fraction of the low-Km enzyme that is particle bound at the high protein concentrations occurring in situ. PMID- 6291934 TI - Adrenocorticotropic hormone and alpha-melanocyte-stimulating hormone induce secretion and protein phosphorylation in the rat lacrimal gland by activation of a cAMP-dependent pathway. PMID- 6291936 TI - A 500-MHz proton-magnetic-resonance study of several fragments of the carbohydrate-protein linkage region commonly occurring in proteoglycans. AB - The proton-magnetic-resonance spectra of three partial structures of the carbohydrate-protein linkage region that frequently occurs in proteoglycans, namely, beta-D-Galp-(1 leads to 3)-beta-D-Galp-(1 leads to 4)-beta-D-Xylp-(1 leads to O)-L-Ser, were recorded in 2H2O at 500 MHz; they could be completely interpreted, both for the glyco-serines and for the corresponding glyco-xylitols. The chemical shifts and the coupling constants were refined by computer simulation of the spectra. The change in the chemical shift of H-4 of a D galactopyranosyl residue upon substitution at C-3 by a beta-D-galactopyranosyl group is proposed to be characteristic for this particular attachment, making H-4 of galactose a structural-reporter group. The three constituting monosaccharides adopt the 4C1 (D) ring conformation. The terminal galactopyranosyl group and the internal galactopyranosyl residue differ as to the population of rotamers around the C-5/C-6 axis. Concomitantly, the flexibility of their glycosidic linkages is distinct. PMID- 6291937 TI - NMR redox studies of Desulfovibrio vulgaris Cytochrome c3. Electron transfer mechanisms. AB - The 300-MHz proton NMR spectra of the tetrahaem cytochrome c3 from Desulfovibrio vulgaris were examined while varying the pH and the redox potential. The analysis of the complete NMR reoxidation pattern was done taking into account all the 16 redox states that can be present in the redox titration of a tetra-redox-center molecule. A network of saturation transfer experiments performed at different oxidation stages, between the fully reduced and the fully oxidized states, allowed the observation of different resonances for some of the haem methyl groups. In the present experimental conditions, some of the haems show a fast intramolecular electron exchange rate, but the intermolecular electron exchange is always slow. In intermediate reoxidation stages, large shifts of the resonances of some haem methyl groups were observed upon changing the pH. These shifts are discussed in terms of a pH dependence of the haem midpoint redox potentials. The physiological relevance of this pH dependence is discussed. PMID- 6291938 TI - Cell-free synthesis of a larger-molecular-weight precursor of cytochrome c oxidase subunit V from rat liver and the distribution of its mRNA between free and membrane-bound polysomes. AB - Poly(A)-rich RNA from phenol-extracted rat liver polysomes was translated in a heterologous cell-free system derived from wheat germs. The labeled translation products were incubated with an antiserum against cytochrome c oxidase subunit V. After immunoprecipitation and affinity chromatography with protein-A-Sepharose, the isolated antigen-immunoglobulin complexes were analyzed by sodium dodecyl sulfate/polyacrylamide gel electrophoresis and fluorography. Only one protein with an apparent molecular weight of 15 500 was visualized. In immunocompetition experiments with unlabeled individual cytochrome c oxidase subunits IV, V, VI or VII only subunit V could compete with the 15 500-Mr protein synthesized in vitro. Two-dimensional fingerprints of cytochrome c oxidase subunit V and the polypeptide synthesized in vitro showed a high degree of similarity. It is concluded that the cytochrome c oxidase subunit V is synthesized as a precursor with an amino-terminal extension of about 25 amino acids. It was possible to convert the precursor of cytochrome c oxidase subunit V synthesized in vitro to its mature form by intact mitochondria as well as by submitochondrial particles. A chain length of 830 +/- 70 nucleotides was estimated for the poly(A)-rich mRNA of the higher-molecular-weight precursor of rat liver cytochrome c oxidase subunit V. Assuming a molecular weight of 15 500 for the precursor a non-coding region of about 300 nucleotides must exist. In experiments on the site of synthesis it is shown that the poly(A)-rich RNA for the higher-molecular-weight precursor of cytochrome c oxidase subunit V is found in free, loosely and tightly membrane-bound polyribosomes. PMID- 6291939 TI - Light-induced interaction between rhodopsin and the GTP-binding protein. Metarhodopsin II is the major photoproduct involved. AB - We have previously described [H, Kuhn et al. (1981) Proc. Natl Acad. Sci. USA, 78, 6873-6877] a light-induced scattering change ('binding signal') associated with a stoichiometric binding between photoexcited rhodopsin and a peripheral membrane protein, the GTP-binding protein, in bovine rod outer segment suspensions. We have attempted here to identify the rhodopsin intermediate R* which is responsible for this interaction, by studying its dependence on pH, temperature and ionic strength. The results strongly suggest that the active state is metarhodopsin II (M II). 1. The initial phase of the binding signal is slightly slower than the formation of metarhodopsin II (2-37 degrees C, pH 5.5 9). 2. The kinetics of the decay of the active rhodopsin state are similar to those of the metarhodopsin II leads to metarhodopsin III transition (37 degrees C, pH 7.3). 3. All conditions which lead to light-induced binding of the GTP binding protein to R* also lead to the formation of M II. At 2 degrees C, pH 8.3, in particular where no M II is formed in the absence of GTP-binding protein, binding signals and light-induced attachment of the GTP-binding protein to the membrane are still observed. Consistently, addition of GTP-binding protein to a suspension of extracted membranes bleached at 2 degrees C (pH 8.3) shifts the metarhodopsin I in equilibrium metarhodopsin II equilibrium towards metarhodopsin II. The shift is reversed by GTP, which dissociates the rhodopsin--GTP-binding protein complex. 4. At low ionic strength, where the GTP-binding protein is soluble in the dark (instead of being associated to the membrane as in the above experiments) M II still induces the binding whereas M I does not, indicating a much lower affinity of the GTP-binding protein for MI. PMID- 6291940 TI - An RNA-dependent nucleoside triphosphate hydrolase from Krebs-II ascites tumor cells. Detection and preliminary characterization. AB - A novel enzymatic activity, RNA-dependent, NTPase, was isolated from Krebs-II ascites tumor cells. This activity is associated with ribosomes and can be detached from them by washing in KCl solutions of a higher than 0.3 M concentration. The enzyme hydrolyzes all the four nucleoside triphosphates to the corresponding nucleoside diphosphates and orthophosphate. The rate of NTP hydrolysis increases about 10-fold in the presence of natural RNAs and synthetic polyribonucleotides [except poly(G)]. Natural DNAs, both double and single stranded, are poor cofactors, although pol(dA) and poly(dT) stimulate, to a certain extent, the rate of ATP hydrolysis. Possible involvement of RNA-dependent NTPase in protein biosynthesis is discussed. PMID- 6291941 TI - The intracellular concentration of pyrophosphate in the batch culture of Escherichia coli. AB - We applied our colorimetric PPi determination method [Heinonen, J., Honkasalo, S. and Kukko, E. (1981) Anal. Biochem. 117, 293-300] to bacterial cultures and measured the intracellular concentration of PPi in the batch culture of Escherichia coli. The cells growing in minimal medium contained about 2.5 nmol PPi/mg protein (0.5 mM). There was no extracellular PPi. A similar concentration of PPi was found also in cells growing in minimal medium enriched by amino acid mixture. PMID- 6291942 TI - [Development of the stimulation of adenylate cyclase by isoproterenol and beta corticotropin (1-24) during the adipocyte conversion of 3T3-F442A cells in culture. Separate effects of insulin and beta-receptor induction during differentiation]. PMID- 6291943 TI - Relationship between membrane integrity and the inhibition of host translation in virus-infected mammalian cells. Comparative studies between encephalomyocarditis virus and poliovirus. PMID- 6291944 TI - Increased sensitivity of left ventricular relaxation to inotropic interventions in patients with coronary artery disease: comparison with normal subjects, mitral stenosis and asymmetric septal hypertrophy. PMID- 6291945 TI - Clinical evaluation of single photon emission computed tomography of the brain. AB - Single photon emission computed tomography (ECT) was performed on 67 patients. ECT images were taken with a Shimadzu scintillation camera, LFOV-E, before a delayed scan. Eighteen of 67 patients showed abnormal findings on the ECT images. Fourteen of the 18 had a transmission X-ray CT (TCT) study as well. There were eleven cases with brain metastasis, and a surgical wound. Eleven of forty-nine ECT-negative patients had a TCT study as well, and intracranial lesions were found in five. The smallest lesion found by ECT was 0.5 cm in diameter on the TCT image and the largest lesion missed by ECT was a tumor in the corpus callosum, measuring 4.2 X 2.7 cm. As far as the patients who also received TCT study are concerned, both the ECT and the ordinary scan were thought to be equal in sixteen patients and ECT to be superior in seven whereas the ordinary scintigram was superior in two. At present, ECT is considered to be useful when it is used in addition to the ordinary scans. In the field of clinical nuclear medicine, the development of new radiopharmaceuticals which are labeled with single photon emitters and which can show the metabolic activity of the brain is eagerly awaited. PMID- 6291947 TI - Radionuclide angiography of the popliteal arteries in occlusive vascular disease. AB - Before undertaking a femoro-popliteal bypass graft it is important to know whether the popliteal artery is patent to receive the distal end of the graft. Patency was assessed in 48 studies of 46 patients with peripheral vascular disease by radio-nuclide angiography. The procedure consists of imaging the popliteal arteries using a gamma camera after a bolus injection of 99mTc-sodium pertechnetate. The radio-nuclide angiograms were compared with the operative X ray popliteal angiograms. Reliable information was obtained from the radio nuclide images regarding the patency of the popliteal artery provided a femoral pulse was present. Quantitative data generated from the activity time curves were of no value in assessment of the patency of the popliteal arteries. PMID- 6291946 TI - A comparative study of 99mTc and 131I in thyroid scanning. AB - Thyroid scans performed with both 99mTc pertechnetate (99mTcO4) and (131I) were compared in 46 patients with palpable thyroid nodules to determine whether 131I scanning is any longer a necessary procedure. A discrepancy between the two types of scan existed in only three cases, in one of which the thyroid nodule showed uptake of 99mTcO4 but not of 131I. Subsequent surgery revealed a thyroid malignancy in this patient. In each of the other two discrepancies a nodule "cold" on 99mTcO4 scanning was apparently functioning on 131I scanning, and was found to be benign at surgery. As the convenience and lower radiation absorbed dose of 99mTcO4 patients compared with 131I make it a better scanning agent, it is recommended that 99mTcO4 scans of the thyroid be first nodules. If these nodules prove to be functioning equally with paranodular tissue, a 131I scan should also be performed to help exclude a possible thyroid malignancy. PMID- 6291949 TI - Effect of high level bovine parathyroid extracts on serum PTH, cAMP and urinary cAMP, Na+, and uric acid in stone-forming patients. AB - Patients with a history of idiopathic calcium oxalate stones, but without current stone formation, show distinctly higher levels of serum cAMP (cyclic adenosine 3',5'-monophosphate) after rapid injection of an extreme concentration of parathyroid extract (PTE). Extreme parathyroid hormone (PTH) levels, as induced by rapid injection of biologically active bovine PTEs, significantly affect the renal generation of cAMP and the elimination of electrolytes in a healthy control group. The stimulated increase of urinary cAMP, sodium, potassium, calcium and phosphate are observed to return to normal levels 120 min after extreme PTE injections for both groups studied. PMID- 6291948 TI - Focal defect in a liver scintigram. PMID- 6291950 TI - Neuropeptides and psychopathology. PMID- 6291951 TI - Effect of parathyroid hormone, calcitonin and growth hormone on cAMP content of growth cartilage in experimental uraemia. AB - The response of proximal tibial growth cartilage cAMP content to different hormonal stimuli, i.e. parathyroid hormone, calcitonin and somatotropic hormone was evaluated in rats with bilateral or subtotal nephrectomy. In uraemic rats, basal cAMP content of growth cartilage was unchanged. Administration of 1-34 PTH in vivo or incubation of growth cartilage with 1-34 PTH in vitro caused a significantly smaller increment of cAMP in uraemic rats (40 IU PTH in vivo: 11.4 +/- 1.01 pmol cAMP/mg protein; controls 24.0 +/- 2.55; P less than 0.001). This finding implies PTH resistance. Diminished cAMP response in uraemic animals was not changed by pretreatment with 1,25(OH)2D3 or parathyroidectomy. The increment of cAMP content of growth cartilage of uraemic animals was significantly (P less than 0.01) greater after in vivo administration of 10 IU calcitonin (46.1 +/- 4.89 pmol/mg protein; control: 29.0 +/- 3.99) or incubation of cartilage with calcitonin in vitro. This finding implies overresponsiveness to calcitonin. Neither in acute nor in chronic uraemia, STH caused a significant change of cartilage cAMP or cGMP content, but STH stimulated 3H-thymidine incorporation into chondrocytes of rats with 5 days uraemia (solvent 2.98 +/- 0.51 x 10(3) cpm per cartilage; STH 5.08 +/- 0.34; P less than 0.05) and caused significant improvement of longitudinal growth of rats with 20 days uraemia. PMID- 6291952 TI - Opioid dependence and cross-dependence in the isolated guinea-pig ileum. AB - The development of opioid dependence and tolerance attributed to selective types of opiate receptors was studied in the isolated ileum of guinea pigs chronically exposed to specific opioids. These investigations were based on reports that in this preparation highly tolerant opiate receptors may coexist with opiate receptors of almost unchanged sensitivity. Thus, the ilea were set up in vitro and tested for tolerance and dependence. Apparently precipitation of the withdrawal contracture, indicating dependence, proved a more sensitive parameter than the phenomenon of tolerance. Maximal dependence was determined at rather low degrees of tolerance (5 to 10 fold). The intensity of the withdrawal contracture failed to increase as opiate tolerance did. Furthermore, the experiments failed to present evidence for the existence of selective dependence at specific opiate receptor types. These findings may suggest multiple adaptational mechanisms upon chronic activation of opiate receptors. One mechanism may be responsible for the development of dependence and a low degree of tolerance, whilst a further increase of tolerance may be associated with changes at the opiate binding site level. PMID- 6291953 TI - [3H]Rauwolscine and [3H]yohimbine binding to rat cerebral and human platelet membranes: possible heterogeneity of alpha 2-adrenoceptors. AB - The specific binding of the alpha 2-adrenoceptor antagonists [3H]yohimbine and [3H]rauwolscine to membranes prepared from rat cerebral cortex and human platelets was rapid, reversible, saturable and of high affinity. Both ligands labelled an identical population of sites in cerebral or platelet membranes though the affinities of both agents were significantly lower in brain. The specific binding of [3H]yohimbine and [3H]rauwolscine in both tissues was displaced by various adrenergic agents in a manner that suggests that the labelled sites probably represent the alpha 2-adrenoceptor. There were, however, significant differences in the affinities of certain alpha-antagonists between cerebral and platelet preparations, even though these agents generated displacement curves with slopes close to unity in both tissues. Thus, whereas yohimbine, rauwolscine, WB4101 and corynanthine were all weaker in cerebral cortex, prazosin was significantly more potent at cortical than in platelet membranes. A possible heterogeneity of alpha 2-adrenoceptors is discussed. PMID- 6291954 TI - MuMTV antigen expression and mammary tumor incidence in non-inbred dd mice. AB - The expression of mammary tumor virus (MTV) antigen in the milk and various organs of three non-inbred dd mouse stocks (ddO, ddN and ddY) was examined by the immunodiffusion (ID) and micro-immunodiffusion (micro-ID) tests. The rate of MTV antigen expression in the milk was 100% at the first lactation in ddO (6/6) and ddN mice (10/10), and 23% in ddY mice (3/13). Mammary tumor incidence was 13% (mean tumor age: 12.0 months), 32% (9.6 months) and 10% (11.5 months) in ddO, ddN and ddY mice, respectively, In F1 hybrids between MTV-free BALB/c females and dd males, a high level of MTV antigen was detected by the ID test in the milk of (BALB/c X ddO) F1, however, the levels in (BALB/c X ddN) F1 and (BALB/c X ddY) F1 mice were low at the first lactation and elevated with the advance of lactation number. Mammary tumor incidence had a trend to be higher and earlier in these F1 hybrids than in non-inbred dd stocks. The development of mammary tumors and detection of MTV antigen in F1 hybrids indicate the extrachromosomal transmission of MTV by male dd mice. The micro-ID test has shown that the mammary tumors, mammary glands, male genital organs except for the testis and the salivary gland expressed MTV antigen, with a high frequency of suggesting that secondary male genital organs may play an important role in MTV infection in mice. PMID- 6291955 TI - [Histochemical study of glycosaminoglycan in Wilms tumors of rats and rabbits and fetal kidneys of rats]. AB - Histochemical studies were performed on glycosaminoglycan (GAG) components in Wilms tumors of rats and rabbits and fetal kidneys of rats. 1. Wilms tumors. The interstitial components had an intense stainability with Alcian blue, and digestion test with hyaluronidase and chondroitinase was intensely positive. The epithelial tumor cells, showing a tubular pattern, were also stained with Alcian blue. The enzyme-digestion test was positive in the basal portion of the tumor cells, whereas it was negative in the free surface of the cells. 2. Rat kidneys. In fetal rats, the epithelial cells as well as the interstitial components were stained with Alcian blue, and the enzyme-digestion test was intensely positive. However, in adult rats, Alcian blue-stainability in the free surface of the tubular epithelium was not abolished by treatment with hyaluronidase and chondroitinase. The substance in the free surface seemed to be consisted mainly of heparan sulfate. Although the physiological significance of GAG in the cell surface cannot be deduced, it is quite conceivable that GAG, especially heparan sulfate, plays an important role in fluid absorption and transportation. PMID- 6291957 TI - Natural transmission of Entamoeba histolytica from mother cynomolgus monkeys (Macaca fascicularis) to their newborn infants under indoor rearing conditions. AB - Natural transmission of Entamoeba histolytica from infected mother cynomolgus monkeys (Macaca fascicularis) to their newborn infants was investigated by the stool examination under indoor individually-caged conditions. Every infant became positive for E. histolytica between 5 and 10 weeks after birth. Although some infants were demonstrated to harbor the trophozoites, no overt amebic diarrhea occurred in any of the infants throughout the period of this study. PMID- 6291956 TI - Body weight changes of mice with different immunity to Sendai virus after challenge infection. AB - Four groups of mice immunized with Sendai virus in different ways, namely convalescent (CT), intrapertoneally (IP) or intranasally (IN) immunized with the inactivated virus and non-immunized (NOI), were challenged and examined degree of immunity and body weight changes. Degree of immunity assessed based on the antibodies, the lung lesions and the lung viral titers was higher in order of CT, IP, IN and NOI. Body weight changes, expressed by both the decrease in mean body weights and the correlation coefficients between pre- and post-challenge, were less in the mice with higher immunity against Sendai virus. PMID- 6291958 TI - [An attempt for eradication of respiratory diseases in breeding mice by sanitary improvement of care]. AB - An attempt was made to eradicate respiratory diseases developed in about 1,000 mice of 19 congenic inbred strains which were maintained in a mouse breeding room. The contagious diseases with respiratory signs were found to be caused by mixed infections with Mycoplasma pulmonis and Sendai virus. The eradication of the diseases was mainly made by sanitary improvement in care of the mice such as intensive disinfection, culling some diseased mice and so on, instead of destroying all colonies. As the result, mycoplasma infection decreased gradually, resulting in a complete eradication about one and half years later and remarkable increases in litter size and weaning rate of mice were obtained. Sendai virus infection failed to be eradicated. PMID- 6291960 TI - HVJ (Sendai virus)-induced envelope fusion and cell fusion are blocked by monoclonal anti-HN protein antibody that does not inhibit hemagglutination activity of HVJ. PMID- 6291959 TI - Gangliosides as receptors for fibronectin? Comparison of cell spreading on a ganglioside-specific ligand with that on fibronectin. PMID- 6291961 TI - The effects of a collagenous extracellular matrix on fibroblast membrane organization. An ESR spin label study. PMID- 6291962 TI - Differential effects of various cAMP derivatives on the morphological and electrical maturation of neuroblastoma x glioma hybrid cells. PMID- 6291963 TI - Loss and restoration of glucagon receptors and responsiveness in a transformed kidney cell line. AB - A kidney cell line (MDCK) retains an adenylate cyclase system sensitive to glucagon, vasopressin, isoproterenol and prostaglandin E1. The stimulatory effect of glucagon on cAMP production was selectively lost in a cloned line derived from MDCK cells transformed by Harvey murine sarcoma virus. Sensitivity to glucagon was largely restored by treatment of the transformed cells with prostaglandin E1 or butyrate. Loss and reappearance of glucagon receptors seemed to be responsible for the observation. The parental MDCK line produced prostaglandins and in the transformed line, this function was abolished. These observations suggest that synthesis of glucagon receptors is controlled by endogenously produced prostaglandin in MDCK cells and that loss of glucagon receptors and their responsiveness in the transformed cells occurs as a consequence of the inability of these cells to synthesize this prostaglandin. PMID- 6291964 TI - Tubuloreticular structures in S49 cells. Relation to cAMP-dependent protein kinase. AB - Tubuloreticular structures (TRS) occur spontaneously in S49 mouse lymphoma cells grown in suspension culture. These structures appear in approx. 20% of the cells in electron microscopical cross-sections. Cloning of S49 cells in semi-solid agarose reveals that TRS are potentially present in all S49 cells. An increase of 60% in the frequency of TRS was observed following exposure of S49 cells to 2 X 10(-4) db-cAMP. This increase was not observed in mutant S49 cells lacking cAMP dependent protein kinase activity. The data suggest that one possibility for the regulation of TRS is through a pathway involving cAMP-dependent protein kinase. In addition, TRS also appear in tumors derived from S49 cells and therefore serve as an ultrastructural cytoplasmic marker for these cells, both in culture and in syngeneic hosts. We suggest the S49 cells as a model system for studying the regulation and function of tubuloreticular structures in vitro and in vivo in malignant lymphoid cells. PMID- 6291965 TI - Expression of lysosomal enzymes in human mutant fibroblast-chick erythrocyte heterokaryons. AB - The generation of enzymes located in lysosomes, in cytosol or in endoplasmatic reticulum/Golgi complex is studied in heterokaryons in which chick erythrocyte nuclei are reactivated. The lysosomal enzymes, alpha-glucosidase (alpha-glu) and beta-galactosidase (beta-gal), are synthesized in heterokaryons obtained after fusion of chick erythrocytes with human fibroblasts of patients with Pompe's disease (alpha-glu-deficient) and GM1-gangliosidosis (beta-gal-deficient), respectively. The enzymes appear to be of chick origin and their activities can be detected at first around 4 days after fusion, i.e., at a time when the nucleoli in the erythrocyte nuclei have been reactivated. Maximal activities are reached around 15 days after fusion. No generation of the lysosomal enzyme beta hexosaminidase is detected in the heterokaryons up to 23 days after fusion of chick erythrocyte with either beta-hexosaminidase A- and B-deficient fibroblasts (Sandhoff's disease) or beta-hexosaminidase A-deficient fibroblasts (Tay-Sachs disease). Similarly no expression of the cytosol enzyme glucose-6-phosphate dehydrogenase (G6PD) is fond up to 30 days after fusion, when chick erythrocytes are fused with fibroblasts from two different G6PD-deficient cell strains (residual activities of 4 and 20% respectively). Indirectly we examined N-acetyl glucosamine-1-phosphate transferase activity, an enzyme located in the endoplasmic reticulum/Golgi region. This enzyme is needed for the phosphorylation of the lysosomal hydrolases and absence of its activity is the cause of the multiple lysosomal enzyme deficiencies in patients with I-cell disease. The retention of both, chick and human beta-galactosidase in the experiments in which I-cell fibroblasts were fused with chick erythrocytes indicates a reactivation of the gene coding for this phosphorylating enzyme. It also implies that this step in the processing of human lysosomal enzymes is not species-specific. PMID- 6291966 TI - Phosphorylation changes induced by cAMP derivatives in the CHO cell and selected mutants. AB - Phosphorylation analysis of CHO mutants selected for unresponsiveness to the Reverse Transformation reaction of cAMP derivatives has been carried out by 2D gel electrophoresis in hopes of finding mutants differing from the wild-type cell in a minimal number of such phosphorylations. Seven differences in protein phosphorylations in the parental CHO cell have been identified as a result of treatment with db-cAMP. db-cAMP-unresponsive mutants of two kinds have been found: One type has lost all seven of the phosphorylation changes induced by db cAMP in the wild-type cell. The other type which is equally resistant to the Reverse Transformation response differs from the parental cell in only one (or possibly two) phosphorylation(s) involving a 55 000 D protein. This protein may, therefore, be directly related to the events of Reverse Transformation. PMID- 6291967 TI - Glycoproteins of Sendai virus (HVJ) have a critical ratio for fusion between virus envelopes and cell membranes. AB - The biological activity of two glycoproteins, hemagglutinin and neuraminidase (HN) and fusion (F) proteins, of Sendai virus (HVJ) were studied using purified proteins. The proteins were purified by chromatography on DEAE and CM cellulose in the presence of Nonidet P-40 (NP40). The glycoproteins were reconstituted at various ratios of F to HN into lipid vesicles containing fragment A of diphtheria toxin. The association of HN and F proteins with the vesicles was confirmed by electron microscopy and sucrose density gradient centrifugation. The cytotoxic activity of vesicles containing fragment A on fusion with L cells was determined by measuring colony formation of the cells. It was found that for maximum cytotoxic activity of the vesicles, there was an optimal ratio of F to HN of two. This suggests that HN is not merely the initial binding site to the cell surface, and that interactions between HN and F proteins on the virus surface may be important for the biological activities of these proteins on the cells. PMID- 6291968 TI - Biochemical and immunological characterization of Na+, K+-ATPase from lens membrane and other tissues. PMID- 6291969 TI - Feline leukemia: a naturally occurring cancer of infectious origin. PMID- 6291970 TI - Modulation of murine in vitro erythroid and granulopoietic colony formation by ouabain, digoxin and theophylline. AB - Ouabain has been shown to increase the number of clonally derived erythroid stem cells, CFUE and BFUE, from normal murine bone marrow. We report here that digoxin and theophylline also enhance erythroid stem cell colony formation, in the presence of suboptimal concentrations of erythropoietin (Ep) (0.01 IU/ml) in methylcellulose culture. Both digoxin and theophylline increased CFUE colony formation optimally at 10(-8) M (29-81% respectively). The increase in BFUE colony formation occurred at 10(-10) M (35-76% respectively), suggesting that BFUE are more sensitive to the enhancing properties of these compounds. In addition, digoxin theophylline and ouabain were effective inhibitors of clonally derived granulocyte-macrophage progenitor cells, CFUC, from normal murine marrow plated in double layer agar in the presence of 10% L-cell conditioned medium (LCM). The degree of reduction in colony formation by CFUC ranged from 11% to 100%. Digoxin and theophylline were inhibitory for CFUC in the range of 10(-2) to 10(-12) M; however, ouabain was inhibitory over a broader concentration range of 10(-4) to 10(-18) M, suggesting that ouabain has a greater influence on committed hematopoietic progenitor cell colony formation. Both ouabain and digoxin have the property of binding to Na+/K+ATPase. This may mediate the alteration of hematopoietic differentiation. Theophylline, an adenyl cyclase inhibitor, may act through alterations in cyclic nucleotide levels. These studies further indicate that digoxin, theophylline and ouabain may serve as useful tools in elucidating the underlying mechanisms of how specific growth factors influence hematopoietic growth and differentiation. PMID- 6291971 TI - Tremor measurement in asthma. II. Changes after terbutaline administration suggesting beta-adrenergic blockade. AB - Repeated finger tremor measurements have been performed in six normal and eight asthmatic subjects after subcutaneous and oral administration of terbutaline. Prior to the investigations the patients had not used bronchodilators for 10-14 days. The finger tremor pattern showed a striking parallelism with the terbutaline serum concentrations and with the cAMP plasma levels. After subcutaneous administration the increase in tremor values in asthmatic subjects was significantly less than in normals. This suggests a beta-blockade in skeletal muscle of asthmatics. The difference after oral administration was not statistically significant. PMID- 6291972 TI - Effects of glucocorticoids on the airways. AB - Glucocorticoids can affect every stage of inflammatory and immunological reactivity. They influence the movement and distribution of lymphocytes, neutrophils and eosinophils in quite low concentrations and decrease the accumulation of these cells at inflammatory sites. The functional capabilities of these cells are relatively resistant to steroids. Steroids also inhibit the leakage of fluid and cells from capillary beds. Glucocorticosteroids have recently been shown to stimulate the synthesis of a protein called lipomodulin or macrocortin, which inhibits the activity of phospholipase A2. As a consequence of this effect the phospholipid methylation in the cell membrane is inhibited, as is the arachidonic acid cascade. This decreases neutrophil and macrophage chemotaxis, histamine release from mast cells and basophils as well as bronchospasm and inflammatory oedema mediated by leukotrienes. PMID- 6291974 TI - Corticospinal neurones of the supplementary motor area of monkeys. A single unit study. AB - The direct projection from the supplementary motor area (SMA) to the spinal cord was investigated in six monkeys by means of antidromic identification of single SMA neurones. The exploration concentrated on that portion of medial area 6 from which movements were found to be elicited by stimulation at intensities of less than 30 microA in an earlier study, but also included some of medial area 4. Of 315 identified corticofugal projection neurones, 234 were found to be localized within medial area 6; of these only one third (76 cells) were corticospinal cells and the remaining two thirds were neurones which projected to the brainstem. The conduction velocities of the descending projection neurones of the SMA were slow (modal value: 10 m/s). Corticospinal cells of the SMA were found up to 6 mm rostral to the boundary between areas 4 and 6. Corticospinal neurones activated antidromically from the cervical but not from the lumbar cord ('cervico-thoracic' neurones) were concentrated in the mesial cortex; 'lumbo-sacral' neurones were found both in the dorsal cortex and the dorsal bank of the cingulate sulcus. However, there was considerable intermingling between the two types of projection neurones and there was no separation in the rostro-caudal direction. Similarly, projection neurones receiving orthodromic inputs from the somatotopical subdivisions of the precentral cortex were not segregated, but were intermingled in the entire rostro-caudal extent of the SMA. It is concluded that there is a clustering of corticospinal neurones in the SMA according to their most caudal segmental projection. However, no rostro-caudal differentiation into face, arm and leg areas was established. This observation is consistent with the results of a previous study in which corticospinal neurones in the SMA were labelled with anatomical tracers and efferent zones were investigated with intra-cortical microstimulation (Macpherson et al. 1982). PMID- 6291975 TI - Autoradiographic demonstration of retinal projections to the brain stem structures in the rabbit using transneuronal tracing technique with special reference to the retinal projections to the inferior olive. PMID- 6291973 TI - Roll tilt reflexes after vestibulospinal tract lesions. AB - The effects of lesions of the vestibulospinal tracts on vestibular reflexes evoked by roll tilt in forelimb and neck extensors were examined in decerebrate cats. Sectioning the medial longitudinal fasciculus, which contains the medial vestibulospinal tract, had no major effect on the phase of the reflex, although some gain was usually lost at high stimulus frequencies. Spinal lesions at C2-C3, both cord hemisections and more restrictive lesions which cut the lateral vestibulospinal tract, produced two major effects on the forelimb. Background EMG activity was usually abolished in the triceps ipsilateral to the lesion, with partial loss of activity in the opposite limb. The tilt reflex response in the ipsilateral limb appeared normal, although it was usually necessary to raise the background excitability of the preparation by administering L-Dopa in order to observe the reflex. In contrast, the response in the contralateral limb showed a phase reversal of 180 deg at low stimulus frequencies, implying that the reflex in intact cats receives a crossed otolith-spinal input. Responses in the neck extensors splenius and biventer, recorded from compartments caudal to the spinal lesion, were relatively unaffected. PMID- 6291976 TI - Participation of projections from substantia nigra reticulata to the lower brain stem in tuning behavior. PMID- 6291977 TI - Prevention of the organophosphorus neuropathy by glucocorticoids. PMID- 6291978 TI - Development and reduction of synaptic potentiation induced by perforant path kindling. PMID- 6291979 TI - Cyclic AMP concentration in the rat's preoptic region. PMID- 6291980 TI - Apparent equilibrium constant of the hypoxanthine guanine phosphoribosyltransferase-catalyzed IMP-GMP exchange. PMID- 6291981 TI - Clinical trials in West Germany: adjuvant therapy for breast cancer patients. PMID- 6291982 TI - [Effect of imizin and transamine on the bioelectrical activity of the ventrolateral columns of the spinal cord]. AB - It was established in experiments on spinal rats that imizine (2.5-10 mg/kg) decreased spontaneous activity and that evoked by stimulation of the sciatic nerve and intraarterial injection of bradykinin in the dorsal horns and ventrolateral columns of the spinal cord. Transamine (1-5 mg/kg) reduced the evoked activity in the dorsal horns and ventrolateral columns but potentiated spontaneous activity in these structures. Naloxone (1 mg/kg) reduced the effect of imizine (5 mg/kg) on the bradykinin-induced nociceptive activity in the ventrolateral columns of the spinal cord. PMID- 6291983 TI - [Effect of delta 9-tetrahydrocannabinol on the cat's differentiation of visual signals]. PMID- 6291984 TI - Kinetics and sidedness of ubiquinol-cytochrome c reductase in beef-heart mitochondria. PMID- 6291985 TI - Decreased guanine nucleotide binding and reduced equivalent production by brown adipose tissue in hypothalamic obesity. Recovery after cold acclimation. PMID- 6291986 TI - Synthesis and metabolism of leukotriene B4 in human neutrophils measured by specific radioimmunoassay. PMID- 6291987 TI - Platinum complexes inhibit ATP-driven basic dye uptake in lysosomes from rat liver. PMID- 6291988 TI - Studies on the desensitization of the cyclic AMP response to thyrotropin in thyroid tissue. PMID- 6291989 TI - Identification and genomic organization of human tRNALys genes. PMID- 6291990 TI - Quantification of the role of the adenine nucleotide translocator in the control of mitochondrial respiration in isolated rat-liver cells. PMID- 6291991 TI - The effect of superoxide generation on the ability of mitochondria to take up and retain Ca2+. AB - When heart or liver mitochondria are exposed to superoxide radicals generated from xanthine + xanthine oxidase their ability to take up and to retain Ca2+ is impaired. The rate of oxidation of pyruvate + malate as substrates is diminished and the appearance of thiol groups when the mitochondria are supplied with these substrates is abolished. These inhibitory effects are offset if respiration is supported by succinate in presence of rotenone provided that a substrate (beta hydroxybutyrate) is provided to maintain the reduction of NADH. The data agree with the thesis that a generation of thiol groups is essential to maintain membrane integrity and that the generation depends on provision of reduced NAD(P)H. PMID- 6291992 TI - Isolation of a heat-stable protein activator of phosphorylase phosphatase. PMID- 6291993 TI - A new method to study permeation of beta-lactam antibiotics into reconstituted vesicles from the outer membrane of Pseudomonas aeruginosa NCTC 10662. PMID- 6291994 TI - Effects of fasting on hepatic catecholamine receptors. PMID- 6291995 TI - The inhibition of proton translocation in the mitochondrial bc1 region by dicyclohexylcarbodiimide. PMID- 6291996 TI - Identification of N- and C-terminal corticotropin peptides in the Mr 80000 form of neurophysin. AB - The 125I-labeled Mr 80000 form of neurophysin has been purified from bovine neurohypophysi. Tryptic digests of this species were analyzed, prior to or after treatment with carboxypeptidase B, by high-pressure liquid chromatography followed by isoelectric focusing and the fragments compared with those generated by a similar treatment of reference bovine 1-39 adrenocorticotropin. The ACTH peptides 22-39 and 1-8, as well as the 1-7 derivative of the latter were identified by those two independent criteria. This provides chemical evidence supporting the hypothesis [8] that high Mr neurophysin may contain the sequence of ACTH. PMID- 6291997 TI - Isolation of a GABA receptor from bovine brain using a benzodiazepine affinity column. PMID- 6291998 TI - Analysis of hepatic growth hormone binding sites of pregnant rabbit crosslinked to 125I-labelled human growth hormone. PMID- 6291999 TI - The mitochondrially made subunit 2 of Neurospora crassa cytochrome aa3 is synthesized as a precursor protein. PMID- 6292001 TI - Growth factors and their receptors in normal and transformed cells: introduction. PMID- 6292000 TI - Intracellular modulation of membrane channels by cyclic AMP-mediated protein phosphorylation in peptidergic neurons of Aplysia. AB - The peptidergic bag cell neurons of the opisthobranch mollusc Aplysia control egg laying and its correlated behavior by release of the neuroactive peptide, egg laying hormone, during the extended electrical discharge termed afterdischarge. This paper examines the evidence for the involvement of cyclic AMP (cAMP) and protein phosphorylation in the mediation of this electrical afterdischarge. It is concluded that an important component in the mechanism of afterdischarge is the suppression of a potassium channel, mediated by cAMP-dependent protein kinase induced protein phosphorylation. The exact identity of the potassium channel remains to be worked out. PMID- 6292002 TI - Receptor remodeling and regulation in the action of epidermal growth factor. AB - Epidermal growth factor (EGF) initiates a wide variety of events when added to responsive cultured cells. These range from early events requiring only brief exposure to EGF, e.g., stimulation of transport of amino acids or ions, to later events such as commitment of cells to a round of DNA synthesis, a process requiring 6 h or more of continuous exposure to hormone. EGF binding is followed first by phosphorylation of EGF receptors, which can be detected in purified membranes and permeabilized cells, and then by internalization and proteolytic processing of receptors in lysosomes. Native 160,000-dalton EGF receptors contain a site that is not exposed on the cell surface and is highly sensitive to cleavage by an endogenous protease, which yields a 145,000-dalton receptor fragment that retains phosphate acceptor activity. Cleavage of receptor at a trypsin-sensitive site, also not exposed to the cell surface, yields a 115,000 dalton fragment that binds EGF, but contains no phosphorylated species. The data indicate that the phosphate acceptor sites on EGF receptors are localized on a 45,000-dalton cytosolic region. PMID- 6292003 TI - Sarcoma growth factor and other transforming peptides produced by human cells: interactions with membrane receptors. AB - Transforming growth factors (TGFs) stimulate cells to divide in monolayer cultures and to form colonies that grow progressively in soft agar. TGFs are a family of polypeptide hormones that, in vitro, confer on fibroblasts and epithelial cells properties associated with the transformed phenotype. They have been isolated from the supernatant fluids of several human and animal carcinoma and sarcoma cells. TGFs interact with epidermal growth factor (EGF) membrane receptors. They are not detectable in culture fluids from cells that contain high numbers of free EGF cell membrane receptors. One TGF is sarcoma growth factor (SGF), which is released by murine sarcoma virus-transformed cells. Studies have shown EGF and SGF to be two distinct growth factors despite the fact that SGF exerts its effects by specifically interacting with EGF receptors. Addition of SGF to normal indicator cells results in expression of the transformed phenotype. The effects of SGF are reversible; the cells resume their normal growth pattern when the growth factor is removed. Three different human tumor cell lines in culture, a rhabdomyosarcoma, a bronchogenic carcinoma, and a metastatic melanoma, release TGFs that also confer the transformed phenotype on normal fibroblasts. One would expect that, as research into this area continues, new TGFs and their interaction with different specific cell membrane receptors will be described. PMID- 6292004 TI - Growth factors, growth factor receptors, and cell cycle control mechanisms in chemically transformed cells. AB - Previous studies have shown that the chemically transformed AKR-MCA and C3H/MCA 58 cell lines spontaneously arrested growth at high saturation density in the G1 phase of the cell cycle because of depletion of low-molecular-weight nutrients (amino acids and glucose) from the medium. The nontransformed parent cell lines, AKR-2B and C3H/10T1/2, spontaneously arrest growth in G1 at low saturation density because of depletion of essential serum growth factors. If prevented from becoming deficient in growth factor by maintenance in medium with mitogens such as epidermal growth factor (EGF), fibroblast growth factor, or the tumor promoter 12-O-tetradecanoylphorbol-13-acetate, the nontransformed AKR-2B cells behave similarly to the transformed cells and arrest growth in G1 at high saturation density because of nutrient deficiency. This suggests that a major difference between the nontransformed and chemically transformed cells is an inability of the latter to achieve growth factor deficiency. In addition, the transformed AKR MCA and C3H/MCA-58 cells show greatly diminished detectable EGF receptors. These observations could be accounted for by the endogenous production of the response to a growth factor-like substance by the transformed cells. Preliminary data indicate that these chemically transformed cells release a transforming growth factor (TGF) into serum-free medium. Whether TGF is similar or identical to the previously described sarcoma growth factor remains to be established. PMID- 6292005 TI - Evidence for the involvement of beta-endorphin in the human menstrual cycle. AB - The possibility that beta-endorphin, an endogenous opiate, is involved in the regulation of the menstrual cycle was examined. Daily serum beta-endorphin levels, in conjunction with luteinizing hormone, progesterone, and 17 beta estradiol were measured during 26 hormonally normal menstrual cycles. Twenty-one cycles showed a preovulatory peak and postovulatory trough of beta-endorphin, 2 cycles had a postovulatory peak, and 3 had a postovulatory peak with sustained elevation. The raw data were standardized by conversion to "Z-scores," and the composite values were computed for each of the three classes described above. Significance within these three classes was assessed using a one-way analysis of variance with an F-ratio at 95% confidence limits. The composite plot of the 26 cycles showed a statistically significant preovulatory peak occurring 2 days prior to the luteinizing hormone surge and a postovulatory trough of beta endorphin 5 days later. These results suggest that beta-endorphins play a significant role in the neurochemical mechanisms of gonadotropin release. PMID- 6292006 TI - [Functional cell differentiation of vascular smooth muscle and basal vascular tonus]. PMID- 6292007 TI - Failure to confirm the role of immunological tolerance to B (MHC) alloantigens in the growth of RSV-induced tumours in chickens. AB - Chickens of the inbred "regressor" line CB (B12/B12) were made tolerant to the B13 (MHC) alloantigen of the congenic "progressor" line CC and then challenged with Prague strain of Rous sarcoma virus of subgroup C. No changes in tumour growth between tolerant and control groups were observed. These results suggest that there is no cross-reactivity between B13 alloantigen and RSV-induced tumour antigens. PMID- 6292008 TI - [The effect of metoclopramide, a dopamine inhibitor, on aldosterone secretion in patients with primary aldosteronism]. PMID- 6292009 TI - [A study on the mechanism of abnormal steroid secretions in a case of dexamethasone suppressible hyperaldosteronism]. PMID- 6292010 TI - Human Ia-like-antigen-expressed cutaneous T cell lymphoma associated with Epstein Barr virus infection. AB - 2 cases of cutaneous T cell lymphoma are reported. In both cases immunological studies revealed that the tumor cells from the cutaneous lesions had human Ia like antigen on the cell surface, which was considered to be alloantigen primarily present on B lymphocytes. Serum examination showed an extremely high titer of two types of antibody against Epstein-Barr (EB) virus in the 1st case and a relatively high titer of one type of antibody in the 2nd case, suggesting that the patients had EB virus infection. Recently, some hematologists found the significant correlation of T lymphocytosis and the appearance of human Ia-like antigen in infectious mononucleosis. Based on these findings, it is assumed that EB virus has the ability of the immunological transformation of cutaneous neoplastic lymphocytes, and so we should not overlook the susceptibility of phenotypical changes of the original cell by EB virus when the immunological classification of cutaneous malignant lymphoma is done. PMID- 6292011 TI - [Glucagonoma with diabetic ketoacidosis; case report]. AB - Diabetic ketoacidosis is an extremely rare manifestation of glucagonoma. We report such a case in a 72-year-old woman known to be diabetic for seven years. The patient was admitted with diabetic ketoacidosis and associated necrolytic migratory erythrema which suggested the diagnosis of glucagonoma. Plasma glucagon levels were increased (569 to 2298 pg/ml). A vascular tumor of the head of the pancreas without obvious hepatic metastases was visualised by angiography. Duodeno-pancreatectomy including the head of the pancreas led to complete recovery of the mucocutaneous lesions and the plasma glucagon level fell (229 pg/ml). The tumor had several histological characteristics suggesting malignancy and a high glucagon content on extraction. Electron microscopy showed multiple A cells and a few isolated B cells. Most of the cells showed immunoreactivity with anti-glucagon and anti-glicentine antibodies. Three months after surgery, the diabetes was again required treatment with insulin. Plasma glucagon level was again increased and chemotherapy with dimethyltriazenimidazolecarboxamide was undertaken. PMID- 6292012 TI - Restored synergistic entero-hormonal response after addition of dietary fibre to patients with impaired glucose tolerance and reactive hypoglycaemia. AB - Although some dietary fibres (DF) improve glucose tolerance by slowing carbohydrate absorption, other mechanisms are certainly involved. Some of the entero-hormonal responses after DF were investigated in six patients with impaired glucose tolerance and reactive hypoglycaemia. All patients received two different breakfasts, each containing 25 g of starch supplied either as white bread (WB) or a fibre-enriched bread preparation (FB): 4 g hemicellulose and 4 g guar. Metabolic and hormonal responses were evaluated over 5 hours. Compared to WB, the FB had a blunting effect on the resulting blood glucose peak (116 +/- 9 mg/100 ml with FB vs. 148 +/- 15 with WB P less than 0.025) or trough (88 +/- 3 mg/100 ml with FB vs. 79 +/- 5 with WB), and upon the insulin response at 60 min (20 +/- 4 micro U/ml with FB vs. 70 +/- 20 with WB). Gut glucagon immunoreactivity was diminished with FB at 90 min (185 +/- 39 vs. 242 +/- 42 P less than 0.05) and 150 min (180 +/- 39 vs. 242 +/- 40). Pancreatic glucagon was initially similar after FB and WB, but a significant rise was observed with FB at 180 min (116 +/- 17 pg/ml vs. 67 +/- 18 P less than 0.05). The improvement of the blood glucose pattern with DF, especially the suppression of reactive hypoglycaemia, seems to depend partly on reduced and delayed response of the entero-hormonal axis. This in turn results in a better synergistic secretion of insulin and glucagon in the late post-prandial period. PMID- 6292013 TI - Opiate receptors and the metabolic response to intravenous glucose. AB - The role of opiate receptors in the metabolic response to an intravenous glucose load was determined in eight non-diabetic subjects (four of whom showed a positive chlorpropamide alcohol flush response and four who did not). Subjects were studied in a double blind randomised fashion receiving either a saline control or the specific opiate receptor antagonist, naloxone (0.4 mg/min), as an infusion for 5 minutes before and 20 minutes after an intravenous bolus of glucose (0.5 g/kg body weight). Naloxone decreased the early plasma glucose peak in all subjects by increasing the distribution volume but did not alter the fractional glucose clearance. Insulin and glucagon responses to glucose were not altered by naloxone. Naloxone delayed the normal post-glucose rise in the levels of the gluconeogenic precursors alanine, lactate, pyruvate and glycerol suggesting a delay in the usual inhibition in gluconeogenesis following a glucose load. There was no difference in the metabolic response between those subjects who were liable to chlorpropamide alcohol flushing and those who were not either with or without naloxone. We conclude that opiate receptors may influence distribution volume and gluconeogenesis but do not play a major role in either insulin or glucagon secretion or in glucose disposal following an intravenous glucose load. PMID- 6292014 TI - Studies on the toxicity and binding kinetics of abrin in normal and Epstein-Barr virus-transformed lymphocyte culture-I. Experimental results - 4. AB - The effects of treatment with varying doses of abrin, a D-galactose binding lectin, on DNA and protein synthesis of normal and Epstein-Barr virus (EBV) transformed lymphocytes have been previously investigated. Using data on EBV transformed lymphocyte cell density as a function of both time and dose of abrin, the authors introduced the concept of self- and cross-coupling metabolic variables as a means of understanding how abrin affected DNA and protein uptake. In this paper, the self-coupling constant is studied in more detail and the relationship between DNA and protein synthesis is further expanded. We find that there is a significant linear relationship between DNA and protein synthesis in normal lymphocyte culture as measured by abrin interaction in the culture. We further find that there is a much stronger relationship between these variables in EBV-transformed lymphocyte culture. This relationship is further examined, and possible analytic equations are expressed. PMID- 6292015 TI - Development and persistence of receptor 'memory' in a unicellular model system. AB - A single exposure of Tetrahymena to diiodotyrosine stimulated replication of the unicellular organism significantly relative to the control. The stimulatory effect tended to decrease with progressing time, but was still demonstrable after as many as 500 generations. Reexposures to diiodotyrosine also enhanced cell growth, indicating the existence of a receptor 'memory' in respect to the initial exposure, but their effect tended to decline soon after initial stimulation, and did not, in all probability, contribute to the preservation of the 'memory' which itself tends to vanish gradually in due course. PMID- 6292017 TI - Isolation of the homogeneous constituents from non-diffusible sugar-peptide (NSP) fraction originating from bovine plasma--II. The influence on cell growth of BHK 21, BHK/RSV and PR-RSH cells in vitro. AB - 1. The sugar-peptide aggregate isolated from the NSP fraction of bovine blood, added in concentration 20 microgram/ml, decrease of [3H]thymidine incorporation and cell proliferation of BHK-21 and BHK/RSV cells in vitro to approx. 25% of untreated control. 2. Two of four peptide constituents of the aggregate show distinct and extremely different effects on cell growth in vitro. The effects of peptide D mol. wt 5000-6000 is characteristic for peptide growth factors, while the peptide G mol. wt 1200-1800 decreases rapidly the growth of all investigated kinds of cells to 8-16% relative to untreated control. 3. The NSP fraction contains also two non-peptide constituents, which inhibit the growth of Rous sarcoma virus transformed cells to a statistically significant degree and have no effect on the growth of BHK-21 cells. PMID- 6292016 TI - Characterisation of the electron transport chain of an obligate methylotroph, strain 4025. AB - 1. The obligate methanol-utilising bacterium strain 4025 contains cytochromes b and c. Cytochrome a is never present. 2. The soluble cytochrome c is similar to that from other methylotrophs in reacting (slowly) with carbon monoxide and it can be separated into two types, differing markedly in their isoelectric points. 3. Some of the cytochrome b reacts rapidly with carbon monoxide and is thus the likely cytochrome oxidase (cytochrome o). 4. The partially purified, NAD+ independent methanol dehydrogenase is similar to such enzymes from the other methanol-utilising bacteria in respect of its prosthetic group, dependence on ammonia or methylamine for activity and its wide substrate specificity. 5. The fluorescence seen in colonies of this organism is probably due to a flavin derivative. 6. This study of electron transport components does not shed any light on the unusually high copper requirement shown by this methylotroph. PMID- 6292018 TI - Properties and activities of phosphoribomutase in the human leukemic cells. AB - 1. The activities and properties of phosphoribomutase were determined in human leukemic leukocytes and normal polymorphonuclear leukocytes. 2. Leukemic leukocytes had markedly increased phosphoribomutase activity. 3. The Km value of leukemic cells to R-1-P was identical to normal cells. However, the Km value to glucose-1-6-diphosphate in leukemic cells had been shown to be low. 4. Inhibition rates by 2-3-DPG and Pi were lower in leukemic cells. 5. Leukemic cells enzyme was unstable by heating. 6. There were no significant differences in isoelectric points between normal cells and leukemic cells. PMID- 6292019 TI - A Q-cycle mechanism for the cyclic electron-transfer chain of Rhodopseudomonas sphaeroides. PMID- 6292020 TI - Transcellular ion currents. PMID- 6292021 TI - Muscle proteinases and their possible roles in muscle growth and meat texture. PMID- 6292022 TI - The phosphorylation of isocitrate dehydrogenase in Escherichia coli. PMID- 6292023 TI - Inactivation and phosphorylation of yeast fructose 1,6-bisphosphatase. PMID- 6292024 TI - Cytochrome b/c complexes with polyprenyl quinol:cytochrome c oxidoreductase activity from Anabaena variabilis and Rhodopseudomonas sphaeroides GA: comparison of preparations from chloroplasts and mitochondria. PMID- 6292026 TI - The spatial pattern of cell-type differentiation in Dictyostelium. PMID- 6292025 TI - The pathophysiology of postmenopausal hot flushes. PMID- 6292027 TI - The phosphorylation of membranal proteins in Dictyostelium discoideum during development. PMID- 6292028 TI - Ecdysterone induces the transcription of four heat-shock genes in Drosophila S3 cells and imaginal discs. PMID- 6292029 TI - Disaccharidase levels of the rat jejunum are altered by dietary fibre. AB - Rat jejunal disaccharidases were measured after feeding high fat fibre-free, high fat pectin, high fat cellulose, high fat galactomannan and high fat tannin diets for 27 weeks. Compared with the high fat fibre-free diet, pectin lowered both lactase and sucrase levels (p less than 0.025). Galactomannan and tannin lowered only the lactase level (p less than 0.05), while cellulose had no significant effect on disaccharidase levels. These data suggest that dietary fibre components have individual and specific effects on jejunal disaccharidase levels, in the presence of a high fat diet. PMID- 6292030 TI - Appendicectomy prevalences in South African adolescents. AB - From questioning 16,939 South African pupils of 16-18 years, in 56 high schools, mean prevalences of appendicectomies in representative segments of ethnic groups were found to be: rural Blacks 0.6%; urban Blacks 0.7%; Indians, 2.9%; Coloureds (Eur-African-Malay), 1.7%; Whites, 10.5%. Percentages in the sexes were similar. Only those of Indian and Coloured pupils appear to be increasing. Blacks and Whites, respectively, have high and low intakes of fibre-containing foods, which are negatively correlated with appendicectomy prevalences. However, although intakes of fibre-containing foods are slightly higher in Indians and Coloureds than in Whites, the former's appendicectomy prevalences are lower than would be dietarily expected. PMID- 6292031 TI - Streptozotocin treatment in pancreatic cholera (Verner-Morrison) syndrome. AB - A case of pancreatic cholera (Verner-Morrison syndrome) associated with a pancreatic endocrine tumor and hepatic metastases is presented. VIP and HPP plasma levels, initially elevated, were accurately followed in various conditions: during corticosteroid therapy, after pancreatic tumor excision, during and after streptozotocin therapy (1.5 g/m2) by repeated intraarterial route). Only streptozotocin therapy resulted in a reduction of the stool volume with concomitant decrease in VIP plasma levels. However, the size of the hepatic metastases was unchanged and HPP plasma levels remained elevated. It is suggested that VIP represents the tumoral secretion and HPP a marker of the residual malignant tissue. PMID- 6292032 TI - Isolation and characterisation of insulin secretory granules from a rat islet cell tumour. AB - Density gradient centrifugation techniques, using iso-osmotic colloidal silica suspensions (Percoll), were developed for the isolation of insulin secretory granules from a transplantable rat islet cell tumour. These procedures were readily completed within 7 h and from each animal yielded approximately 1 mg of granule protein. The isolated granules were essentially free of other subcellular organelles as evaluated by their contents of marker proteins, electron microscopy and by electrophoretic analyses. Their susceptibilities to lysis at low osmotic strength, at pH values above 7 or in media containing sodium ions were similar to those of granules partially purified from islets. Insulin comprised 50-60% of the total granule protein when determined by immunoassay or by densitometry of electrophoretic profiles. The proinsulin content was marginally higher than that of islets, as was the ratio of insulins I to II. Electrophoretic analyses revealed that the secretory granules contained 150 or more proteins besides insulin-related peptides. The majority of these had acidic isoelectric points and were located both within the granule interior and its enveloping membrane. PMID- 6292034 TI - Hepatotoxic potential of ketoconazole under investigation. PMID- 6292033 TI - The partial protective effect of the hydroxyl radical scavenger dimethyl urea on streptozotocin-induced diabetes in the mouse in vivo and in vitro. AB - The protective effect on streptozotocin-induced diabetes of dimethyl urea, a hydroxyl radical scavenger, has been evaluated in vivo and in vitro. Pretreatment with dimethyl urea before a single diabetogenic dose of streptozotocin partially protected NMRI mice from hyperglycaemia, whereas the serum glucose of C57BL/KsJ mice increased during week 2 of observation. When the pancreases of these latter mice were examined histologically, insulitis was found in 15 out of 22 animals. The protective effect of dimethyl urea in the NMRI mice was not due to short-term hyperglycaemia induced by the drug, since pretreatment with glucose did not protect from streptozotocin but potentiated its diabetogenic effect. Dimethyl urea reduced the inhibition caused by streptozotocin on proinsulin biosynthesis of NMRI islets in vitro. It is suggested that streptozotocin-induced diabetes in mice may involve generation of hydroxyl radicals which are toxic to islet B cells. If this immediate cytoxicity is reduced by a scavenger, a more slowly developing hyperglycaemia and an accompanying insulitis may occur in particularly susceptible animals. PMID- 6292035 TI - Inappropriate use of smallpox vaccine. PMID- 6292038 TI - Interaction of the octapeptide of cholecystokinin and gastrin I with bethanechol in the stimulation of feline colonic smooth muscle. PMID- 6292036 TI - Differential features of some unusual biliary tumors. PMID- 6292037 TI - Effect of enteric hormones on sphincter of Oddi and gastrointestinal myoelectric activity in fasted conscious opossums. AB - In this study we evaluated the effect of enteric hormones on sphincter of Oddi and gastrointestinal spike burst frequency in fasted conscious opossums. A chronic model for obtaining multiple long-term recording of sphincter of Oddi and gastrointestinal myoelectric activity was prepared in 12 animals by implanting bipolar electrodes in the gastric antrum, duodenum, sphincter of Oddi, jejunum, and ileum. Subsequently, 8-18 recording session, 6-8 h in length, were obtained in each animal. Each study was preceded by a 15-h fast. After recording two cycles of the interdigestive migratory myoelectric complex, a 60-min infusion of saline, cholecystokinin-octapeptide, pentagastrin, glucagon, or secretin was begun early in the quiescent period of duodenal activity, 10 min after termination of phase 3 of the migratory myoelectric complex activity. Cholecystokinin-octapeptide (10-80 ng/kg . min) and pentagastrin (25-200 ng/kg . min) increased sphincter of Oddi spike burst frequency while converting interdigestive gastrointestinal myoelectric activity to a fed-like pattern of continuous, irregular spike bursts. Glucagon at high doses of 500 and 1000 ng/kg . min depressed sphincter of Oddi and gastrointestinal spike burst activity, whereas lower doses of glucagon and secretin (3-12 CU/kg . h) had no effect. We conclude that in the opossum, relatively low doses of cholecystokinin-octapeptide and pentagastrin stimulate sphincter of Oddi and gastrointestinal spike burst activity while disrupting the cyclic pattern of sphincter of Oddi and gastrointestinal spike burst activity present during fasting. The physiological significance of these findings remains to be determined. PMID- 6292039 TI - The mode of action of carp gonadotropin on the stimulation of androgen production by carp testis in vitro. PMID- 6292040 TI - The effects of prostaglandins, phosphodiesterase inhibitors, and cyclic AMP on ovulation of brook trout (Salvelinus fontinalis) oocytes. PMID- 6292041 TI - The filamentous phage (Ff) as vectors for recombinant DNA--a review. AB - Derivatives of filamentous phage, f1, fd, and M13, useful as cloning vectors are listed, and procedures for their use are reviewed. Methods for growing phage, preparing single- and double-stranded DNA, and cloning are given in the "cook book" form. These procedures minimize the practical problem often associated with filamentous-phage cloning, i.e., deletion of inserts. PMID- 6292042 TI - A physical map of the bioAB region in the lambda bio transducing phage. AB - The center of the pBopA promoter-operator region for the bioABFCD operon is located 1.71 kb clockwise from the att lambda site on the Escherichia coli genome, as determined by the position of the p131 (IS1) insertion. The order of several bio endpoints to the right of p131 is lambda bio267, 122, 169, 74, 1, and 69. The endpoints of the two bio deletions, delta 61 in bioA and delta 3h in bioB, were also determined. PMID- 6292043 TI - Structure of thymidine kinase gene introduced into mouse Ltk- cells by a new injection method. AB - Pricking, a new injection method developed by Yamamoto et al. (1981), can be used to introduce DNA into cultured cells with high efficiency. Closed circular plasmid DNA containing the cloned HSV-TK gene (pTK-1) was introduced by this method and the structure of DNA in stable transformants was examined. In most clones, the introduced DNA was integrated into the mouse genome in a tandemly repeated form. The possibility of multiple integration via mouse middle repetitive sequences was also examined using the chimeric plasmid with TK genes and middle repetitive sequences (pMRTK-1). Digestion with restriction enzymes showed that the middle repetitive sequence used in this experiment had no effect on the efficiency of transformation, suggesting that this sequence is unable to mediate homologous recombination with mouse genomes. PMID- 6292044 TI - Construction of recombinant plasmids containing rat thyroglobulin mRNA sequences. AB - Two plasmids containing rat thyroglobulin cDNA sequences have been constructed and characterized. A plasmid with a 500-bp insert (pRT6) was isolated and identified as thyroglobulin-specific on the basis of the tissue specificity of the inserted sequence and of its ability to retain thyroglobulin mRNA on a nitrocellulose filter. The cDNA insert in pRT6 was subsequently used to screen a rat thyroid cDNA library constructed with large cDNA. A plasmid was found containing a 1700-bp insert. The polarity and the fidelity of the insert is demonstrated by S1 mapping. PMID- 6292045 TI - Precise and nearly-precise excision of the symmetrical inverted repeats of Tn5; common features of recA-independent deletion events in Escherichia coli. AB - The transposon Tn5 contains a unique central region bordered by 1.5-kb inverted repeats. The in vitro deletion of the centre of Tn5, with a restriction endonuclease (XhoI) which cuts within the inverted repeats leads to the production of a palindrome on subsequent ligation. This palindromic region is unstable on subsequent transformation into Escherichia coli (Collins, 1981). Precise excision of the Tn5 region plus one copy of the bracketing 9-bp direct repeat occurred in about one-third of the transformants. The rest of the transformants contain only remnants of the inverted repeat. Sequence analysis indicated that deletion had occurred between short direct repeats. The precise excision of these "nearly precise" excision products continued with high frequency and was found to be affected by mutations that interfere with the normal precise excision of transposons. In a recB, sbcB host precise excision was markedly reduced. A common mechanism is proposed for all recA-independent deletions occurring in E. coli. PMID- 6292046 TI - Lethality of palindromic DNA and its use in selection of recombinant plasmids. AB - A plasmid derived from ColE1 is constructed so that the removal of one restriction endonuclease HindIII fragment allows the ends of the remaining single fragment (the replicator) to be joined, generating a palindromic sequence 2394 bp in length. The circular species thus produced gives rise to transformants of E. coli at very low frequency. Since the palindromic sequence is effectively lethal to a plasmid containing it, the replicator will give rise to more transformants when the restriction fragment originally removed from it is replaced by another. This principle can be exploited to allow the efficient molecular cloning of unselected restriction fragments. PMID- 6292047 TI - The expression of Streptomyces and Escherichia coli drug-resistance determinants cloned into the Streptomyces phage phi C31. AB - Lysogens obtained by infecting Streptomyces albus G with a phi C31-pBR322 chimaeric prophage or its delta W12 deletion derivative had increased tetracycline resistance. The ability of the delta W12 derivative to transduce tetracycline resistance was inactivated by inserting a viomycin resistance determinant (vph) into the BamHI site of the pBR322 tet gene, and restored by excising the vph gene. Another deletion mutant (delta W17) of the chimaera, carrying an intact tet gene, was normally unable to transduce tetracycline resistance. This inability was correlated with the finding, by Southern hybridisation analysis, that the att site required for insertion of phi C31 prophage into the host chromosome was located within the delta W17 deletion. Use of phi C31 lysogenic recipient permitted the integration of the att-deleted phage, presumably by homologous recombination, giving tetracycline-resistant double lysogens. This technique was extended to S. coelicolor A3(2) in the detection of derivatives of the att-deleted phage into which a thiostrepton resistance determinant (tsr) had been inserted in vitro. Phage released from double lysogens were mainly recombinants. One such recombinant is a PstI vector for DNA cloning, able to accommodate up to 6 kb of introduced DNA. PMID- 6292048 TI - A family of cloning vectors containing the lacUV5 promoter. AB - A family of plasmids containing short pieces of Escherichia coli lac promoter DNA has been constructed. DNA fragments from any source may be inserted directly into the unique EcoRI sites of some of these plasmids to achieve transcription under the control of the lacUV5 promoter. Alternatively, the plasmids serve as convenient sources of lac DNA fragments ('portable promoters') containing the 'up' promoter mutations UV5 or Ps (super promoter) as well as the wild-type promoter. pOP95-2, pOP95-5, pOP203-1, pOP203-2 and pOP203-3 are derivatives of pMB9 while pOP95-15 and pOP203-13 are derivatives of pBR322. The pOP95 plasmids contain the 95-bp AluI lac fragment. This fragment includes the UV5 promoter (minus the CAP binding site), the repressor binding site, and ends 2 bp before an ATG encoding the beta-Gal start codon. The pOP203 plasmids contain the 203-bp HaeIII lac fragment. This fragment contains the UV5 promoter (including the L8 mutation in the CAP binding site), the repressor binding site and sequences encoding the first 8 amino acids of beta-Gal. To shorten and introduce reading frame heterogeneity in the beta-Gal coding end of the pOP203 plasmids, the EcoRI site in pOP203-12 was moved upstream by digesting EcoRI cut plasmid DNA with T4 DNA polymerase and S1 nuclease followed by ligation in the presence of EcoRI linker. This produced the plasmids pOP203-24, pOP203-27, pOP203-28 and pOP203-29. pOP203-29 encodes essentially just that portion of the beta-Gal mRNA sequence which is protected from nuclease digestion by the bound ribosomal complex (Maizels, 1974). PMID- 6292049 TI - The human serum albumin gene: structure of a unique locus. AB - The entire gene for human serum albumin (HSA) has been isolated from a genomic DNA library, carried in the lambda Charon 4A vector. Six independent isolates have been found to hybridize to a cloned HSA cDNA probe, and all six clones share restriction site sequence homology in the overlapping portion of their DNA. These results seem to indicate that the albumin gene is single-copy, or unique, within the human haploid genome. Measuring from the "CAP" site to the "poly(A)" addition site, albumin gene comprises 16.5 kb of DNA. PMID- 6292050 TI - An in vitro method generating base substitutions in preselected regions of plasmid DNA: application to structural analysis of the replication origin of the Escherichia coli K-12 chromosome. AB - A method for introducing base substitutions in defined regions of plasmid DNA has been developed. In principle, a circular heteroduplex DNA containing a gap is constructed by annealing of two kinds of linear molecules derived from the same plasmid: One is the molecule shortened either by exonucleolytic digestion from the termini generated at a restriction site or by removal of a region flanked by two restriction sites, and the other the full-length molecule linearized at a different site. The deleted region in the shorter linear molecule becomes a single-stranded gap in the circular heteroduplex DNA. The heteroduplex is then treated with sodium bisulfite that converts specifically cytosine residues to uracil residues in single-stranded regions. After filling in the gap by repair synthesis, transformation is carried out to isolate mutant plasmids. Since two kinds of circular heteroduplexes are formed by annealing in which the sequences in the gaps are complementary to each other, mutagenesis of both strands can be accomplished in one experiment. This method was applied to construction of mutants with base substitutions in the replication origin region (oriC) of the Escherichia coli K-12 chromosome which had previously been cloned in colicin E1 plasmid vectors, and various mutants in defined regions of oriC were successfully isolated at high efficiencies. Analysis of these mutants provided evidence that oriC contains special regions, designated spacers, which separate neighboring important sequences specifying interactions with initiation factors for DNA replication at precise distances. PMID- 6292051 TI - The nucleotide sequence of the gene encoding protein IVa2 in human adenovirus type 7. AB - The nucleotide sequences of cloned DNA segments encoding the IVa2 gene from Ad7 and a portion of Ad12 (group B and group A human adenoviruses, respectively) have been determined. When compared to Ad5, a group C adenovirus, these sequences have been found to be 80% homologous. Most changes are transitions or transversions. This high degree of nucleotide homology results in a high degree of amino acid conservation in the predicted polypeptides encoded from these genes; most nucleotide changes occur at the third position in the codon. The predicted polypeptide contains 448 amino acids and has a calculated Mr-value of 50700. The positions of the 5' end of the mRNA and of the donor and acceptor splice sites of Ad7 and Ad12 can be inferred by analogy to those of Ad5. A long open reading frame starting upstream from the IVa2 gene overlaps the N-terminal portion of the polypeptide but is encoded in a different reading frame. Within this overlapping region, the long open reading frame is more conserved in amino acid sequence than is the presumed IVa2 polypeptide, suggesting that evolutionary pressure was exerted on the longer protein, a product of viral early region 2B. The high degree of conservation of this E2B region within the overlapping segment suggests that its activities must be more important for adenovirus infection than are the functions encoded in the amino-terminus of the IVa2 gene. PMID- 6292052 TI - Cloning and restriction map of the E. coli apt gene. PMID- 6292053 TI - A physical map of the Escherichia coli bio operon. AB - The endpoints of the Escherichia coli bio DNA insertions in 24 lambda bio transducing phage were mapped electron micrographically in heteroduplexes of the type lambda bio/lambda att2, which permit simultaneous measurement of the lambda deletion and bio insertion endpoints. A physical map of the bio operon was constructed and correlated with the genetic map, the molecular sizes of the bio gene products, and the restriction map. The order att lambda-bioA-pBopA-bioBFCD uvrB was confirmed. The maximum size of the bio operon was estimated at 5.5 kb, and the locus was found to be fully saturated with genes. There appears to be little space between the bioA gene and att lambda, while bioD mapped within 0.7 kb from uvrB. The size of the uvrB locus was estimated not to exceed 2.6 kb. PMID- 6292054 TI - Galactosemic enzyme levels in presenile cataracts. AB - Galactose-1-phosphate uridyl transferase and galactokinase activities have been measured in the red blood cells of a group of patients with "idiopathic" presenile cataract and of a group of nondiabetic patients with senile cataract. The activity of both galactosemic enzymes was found to be within the normal range in all the patients with presenile cataract. In the group of patients with senile cataract, galactokinase activity was normal in all 24 subjects examined, and galactose-1-phosphate uridyl transferase activity was moderately reduced in 3 of 14. PMID- 6292055 TI - [Comparative characteristics of the adsorption properties of natural sorbents in relation to influenza viruses]. PMID- 6292056 TI - [Beta subunit of chorionic gonadotropin in the control of benign trophoblastic disease]. PMID- 6292057 TI - Ultrastructural and functional studies of the platelets in patients with May Hegglin anomaly. PMID- 6292058 TI - Daunorubicin-induced chronic cardiomyopathy--an experimental model system for study of sudden death. A preliminary report. PMID- 6292059 TI - [Possibilities and limits of thyroid sonography in the diagnosis of thyroid cancer]. AB - The echographic appearance of 19 histologically confirmed malignant thyroid tumours has been analyzed. The value of information by ultrasonography is compared to scintigraphy and fine-needle aspiration biopsy. All patients demonstrated a sonographic pattern of relatively low amplitude, sparse and disordered echoes. Extended thyroid tumours (T2-3) exhibited a serrated marginal border as a sign of the infiltration to the surrounding thyroid tissue. The clinically important separation of solid and cystic nodules can always be made with certainty. Provided that a subtile examination technique is used, ultrasonography can be of great value in choosing the adequate therapy in scintigraphic cold nodules. Diagnostic ultrasound also yields information concerning the reliable and reproducible classification and accurate location of circumscribed thyroid lesions. In all cases, however, the ultrasound pattern did not provide reliable clues as to their histology. A successful differentiation between benign and malignant solid nodules can be obtained only by cytology and histology. Ultrasonography, scintigraphy and aspiration biopsy, therefore, are complementary methods. PMID- 6292060 TI - [Value of computer tomography in comparison to mediastinoscopy and test thoracotomy in intrathoracic space-occupying lesions with mediastinal involvement]. AB - The preoperative CT findings in 50 patients out of a total of 144 examined patients with suspected intrathoracal malignant processes with mediastinal involvement were compared to the results obtained by mediastinoscopy and thoracotomy. There was a high diagnostical reliability (95%, n = 40) in assessing lesions in the area, which can be reached by mediastinoscopy. This leads to the conclusion that mediastinoscopy is necessary only in such cases with histologically proven malignant intrathoracal masses, which show lymph nodes of a size from 1 to 2 cm. The evidence of invasive extension into adjacent mediastinal structures as well as the distance from tumour to carina could be assessed well by CT. For follow-up examinations after resection, radiation therapy or chemotherapy, computed tomography is of particular value to detect early mediastinal recurrence. PMID- 6292061 TI - [Malignant fibrous histiocytoma of soft tissue. Pathology and radiology of an unusual sarcoma]. AB - Four cases of malignant fibrous histiocytomas are reported. An attempt is made to determine some characteristic roentgenologic features of this disease. There is no pathognomonic radiologic sign; however, angiographic documentation of hypervascularization together with neovascularity and absorption values of about 35 HE in computed tomography suggest a predominantly fibrous sarcoma. Differential diagnosis on abscess formation, tumours of the fat tissue and primary vascular neoplasms should be performed and these conditions ruled out if not applicable. PMID- 6292062 TI - [Drug therapy in rheumatology]. PMID- 6292063 TI - [Therapeutic possibilities in polyneuropathies]. PMID- 6292064 TI - [Growth characteristics of an attenuated herpes simplex virus type 1 strain SKa in tissue culture cells and in mice]. PMID- 6292065 TI - Effects of acute and chronic endotoxin treatment on glucagon and insulin receptors on rat liver plasma membranes. AB - The effects of acute and chronic endotoxin treatment on the plasma levels of insulin and glucagon and their binding to rat liver plasma membranes were examined. Both acute and chronic endotoxin administration increased plasma glucagon levels and decreased the glucagon to insulin molar ratio. Acute, but not chronic, endotoxin decreased blood glucose and insulin levels. Glucagon binding was increased in membranes prepared from the acutely treated rats. However, in membranes obtained from rats treated chronically with endotoxin, only insulin binding was increased. The increases in the binding of both insulin and glucagon were the result of increases in receptor sites. PMID- 6292067 TI - Interferon production by peripheral lymphocytes in HBsAg-positive liver diseases. PMID- 6292068 TI - Demonstration of albumin receptors on isolated human hepatocytes by light and scanning electron microscopy. AB - The presence of albumin receptors on the plasma membrane of isolated human hepatocytes was investigated employing albumin-coupled latex minibeads. Hepatocyte-latex reaction was visualized by phase contrast and scanning electron microscopy. The experiments demonstrate that hepatocytes exhibit binding activity for polymeric and monomeric forms of glutaraldehyde-treated albumin. Additionally, the reaction was shown to be species-nonspecific. These findings support the hypothesis that polymerized albumin may act as a bridge between receptors on hepatitis B virus and human hepatocytes. PMID- 6292066 TI - Enhancement of renal compensatory hypertrophy by hyperadrenocorticism and its modulation by nutritional factors. AB - Renal compensatory hypertrophy (RCH) is enhanced by ACTH in the uninephrectomized rat. In the present experiments, the kidney weight and its content in protein, RNA and DNA were determined in 48 adult, female rats; 24 had free access to a NaCl solution (9 g/l) and the others to a glucose solution (50 g/l). In each group 12 rats were sacrificed 2 or 7 d. after uninephrectomy (UN). In each subgroup 6 rats were treated with ACTH (18 micrograms/100 g B.W./d) from operation until autopsy. RCH has been evaluated by the arithmetical difference between the data determined in the right control kidney excised at UN and those determined in the left solitary kidney. In all the rats, hyperadrenocorticism increased significantly the weight of the solitary kidney and its content in protein and RNA. There was a significant decrease of the DNA content of the solitary kidney in the rats sacrificed 7 d. post-UN, treated with ACTH and drinking the saline solution. DNA was not affected by ACTH in the 7 other groups suggesting that ACTH favours cellular hypertrophy mainly in the rats drinking the saline solution. The renotrophic action of hyperadrenocorticism may be related to an altered handling of Na+ and K+: there was a positive correlation between the weight gain of the solitary kidney and the urinary excretion of Na+ (r = 0.507, p less than 0.001) and of K+ (r = 0.460, p less than 0.001). Hyperinsulinism was present in all the rats given ACTH; it may act as a growth factor. Hyperglycemia played an important role in former experiments but it was absent in the present studies. PMID- 6292069 TI - Studies on cytochrome c oxidase, IX. The primary structure of polypeptide VIa. AB - The complete amino acid sequence of the cytoplasmic polypeptide VIa of cytochrome c oxidase from beef heart is described. The primary structure of this component of complex IV of the respiratory chain is elucidated by isolation and sequencing of overlapping glutamic acid, arginine, tryptophan and methionine fragments obtained by cleavage with Staphylococcus aureus protease, protease from submaxillaris glands of mice, 2-iodosylbenzoic acid and cyanogen bromide. The chain length of polypeptide VIa is 98 amino acids, the resulting molecular mass of 10670 Da. The hydrophilic protein does not contain a hydrophobic membrane penetrating sequence domain. Its function in the respiratory complex IV is unknown. PMID- 6292070 TI - Inactivation and reactivation of rat liver 3-hydroxy-3-methylglutaryl-CoA reductase phosphatases: effect of phosphate, pyrophosphate and divalent cations. AB - Incubation of four purified rat liver HMG-CoA-reductase phosphatases (Gil, G., Sitges, M. and Hegardt, F.G. (1981) Biochim. Biophys. Acta 663, 211-221) with Mn2 or Mg2 caused a concentration-dependent activation of enzyme activities. The maximum effect for Mn2 was at 5 mM for all phosphatases. Fe2 caused inactivation only in reductase phosphatases IIa and IIb. Ca2 10 mM showed a slight effect of inactivation. Phosphate, pyrophosphate and adenine nucleotides inhibited the four reductase phosphatases, this process being concentration-dependent. cAMP did not inhibit the four phosphatases at all in the range of 0.01-8 mM. Preincubation of reductase phosphatases with PPi and subsequent dilution did not diminish the inactivation effect, showing that this ion inhibits the enzyme prior to the binding to the substrate. Phosphorylated sugars, but not free sugar, inactivated the four reductase phosphatases. PPi-inactivated enzymes were reactivated by Mg2 or Mn2, this process being time-dependent. The four phosphatases had different patterns of reactivation. Phosphatases Ib and IIb (low-molecular mass forms) were shown to be different enzymes as judged by: their divergent behaviour when inhibited with Fe2; their PPi response; kinetics of reactivation by Mg2 or Mn2 or PPi-inactivated enzymes; and thermal stability. A metalloenzyme character is suggested for reductase phosphatases. PMID- 6292072 TI - Tissue-specificity overrides species-specificity in cytoplasmic cytochrome c oxidase polypeptides. AB - With a high-resolving dodecyl sulfate electrophoretic system rat liver cytochrome c oxidase was separated into 13 different polypeptides. An antiserum against rat liver holocytochrome c oxidase immunoreacted with all 13 polypeptides, as demonstrated by immunofluorescence after transfer of the separated Coomassie blue stained bands on nitrocellulose and coupling with FITC-protein A ("western blot"). Polypeptide-specific antisera reacted only with their corresponding polypeptides indicating that the various protein bands are represented by individual polypeptides. From total proteins of rat liver, kidney, heart, spleen and skeletal muscle mitochondria, only the cytochrome c oxidase polypeptides showed immunofluorescence with an antiserum against the rat liver holoenzyme. In contrast to the polypeptide from liver, polypeptide VIa from heart and skeletal muscle showed little or no reactivity, indicating a tissue-specificity of this polypeptide. Mitochondrial proteins from pig, bovine and blackbird heart were incubated with an antiserum against the rat liver holoenzyme. Immunoreaction was found with most cytochrome c oxidase polypeptides but not with polypeptide VIa. This result demonstrates less immunological relationship between tissue-specific polypeptides (VIa, VIIa and VIII) of the same species than between tissue unspecific polypeptides of different species. PMID- 6292071 TI - Dependence of human somatotropin activity on interchain disulfide bridges. AB - The importance of the disulfide bridges for human somatotropin activity is investigated. The activity of somatotropin is tested by the tibia, radioimmuno-, and radioligand assays. The cleavage of disulfide bridges by sulfitolysis, reduction with dithiothreitol, or oxidation with performic acid does not completely abolish hormone activity. There is only one exception: in the radioligand assay, oxidized somatotropin is not able to displace native somatotropin from rat liver membranes. The diminution of hormone activity is independent of the charges of the groups introduced to the cysteine residues. The radioimmuno- and radioligand assays are more sensitive to conformational alterations in the somatotropin molecule than the biological test system. PMID- 6292073 TI - Solitary plasmacytoma of the intestine. PMID- 6292074 TI - Large paraffin sections and chemical clearance of axillary tissues as a routine procedure in the pathological examination of the breast. AB - The technique of large paraffin sections of the breast and lymph node dissection of axillary tissues after chemical clearance as used in a routine histology laboratory is described and discussed. The findings in 155 cases of breast malignancy derived from routine surgical patients and also from the breast screening clinic are described and analysed. There were 21 cases of in-situ carcinoma alone and an additional 17 cases with minimal invasive carcinoma (MIC). The maximum area of each lesion in one plane was measured and the size varied between less than 1 cm2 and 30 cm2, with an average of 7 cm2. The large lesions had the highest incidence of MIC. There were 117 palpable invasive carcinomas and they were divided into two groups, namely single quadrant (80%) and multiquadrant malignancy (20%). The clinical implications of this are discussed. Lymph node metastases were found in 36 (27%) of all invasive carcinomas but no metastases were found in either in-situ carcinoma alone or when combined with MIC in the eight cases which had a radical operation. PMID- 6292075 TI - Benign mixed Mullerian tumour--adenofibroma of the fallopian tube. PMID- 6292076 TI - Effect of growth arrest on the doubling potential of human fibroblasts in vitro: a possible influence of the donor. AB - Population doublings versus time in culture were compared in human postnatal skin fibroblasts from normal donors, a cancer patient, and from donors suffering from Cockayne syndrome, Ataxia telangiectasia, and Fanconi's anemia (FA). Confluent cultures were maintained in a nonproliferating state for 14 to 27 d in 0.5% serum medium. The results show that the ability of cells to resume division after a resting stage can be influenced by pathologic conditions. In arrested FA cell populations an increase of the population doublings and of the calendar time were observed. It is possible that in some cell populations the resting stage favors the expression of growth potentialities related to instability of the cells. PMID- 6292077 TI - Cyclic AMP-dependent protein kinase content of murine splenic T and B lymphocytes and non-lymphoid tissues. AB - Cyclic AMP-dependent protein kinase activity has been measured in DEAE-cellulose elution profiles of highly enriched murine splenic T and B Lymphocytes. B Lymphocytes were found to contain about 10% of the amount of enzyme activity associated with T lymphocytes and several non-lymphoid tissues. Cyclic AMP independent casein kinase activities were equivalent in T and B lymphocytes. Lymphocytes contained predominantly type I isozyme as opposed to several non lymphoid tissues which all contained predominantly the type II form. PMID- 6292078 TI - Adenosine--norepinephrine interactions on cyclic AMP generating system in human tumoral brain slices. AB - Many experimental findings suggest that the administration of exogenous Norepinephrine (NE), in cerebral cortical slices surviving in vitro, increases cyclic AMP (cAMP) levels, although the NE receptor turns out to be different for the different animal species. Likewise, in the same experimental model, Adenosine (A) increases the cAMP intracellular levels. Furthermore, A sites seem to be linked with adrenoreceptors related to the cAMP-generating system. In this report we studied the interaction of NE on the cAMP system of human cerebral tissue slices, normal and tumoral. In the normal slices, NE increases cAMP levels in a dose-dependent manner, probably through beta-receptors; in fact, Propranolol counteracts this effect. Also A induces a dose-dependent rise of cAMP levels; Theophylline prevents, while low doses of Dipyridamole potentiate, this effect. The contemporaneous administration of NE and A produces an effect on cAMP levels greater than that displayed by each drug alone. Probably the enhanced cAMP increase is due to the endogenous release, evoked by NE. In fact, Propranolol reduces, but does not completely prevent, the effect of NE. In cortical tumor slices, the effect of NE on the cAMP-generating system is very reduced. This suggests that the membrane damage of neoplastic cells affects the availability of adrenoreceptors. On the other hand, the responsiveness of A sites is deeply altered. The receptor antagonists or the re-uptake inhibitors of this Adenine nucleotided do not exert their effect selectively. PMID- 6292079 TI - Cytomegalovirus (CMV) infection in children from Kerala state - a preliminary report on serological data. PMID- 6292080 TI - A study of ninety three cases of primary carcinoma of liver. PMID- 6292081 TI - Primary fibrous histiocytoma of the pericardium. PMID- 6292082 TI - Some detoxification methods for N-nitrosamine-contaminated wastes. PMID- 6292083 TI - Hypotensive action of captopril in spontaneously hypertensive and normotensive rats. Interference with neurogenic vasoconstriction. AB - The effects of captopril and angiotensin II on adrenergic neurotransmission have been studied in spontaneously hypertensive rats (SHR) and Wistar-Kyoto rats (WKY). In a pithed rat preparation, vasoconstrictor responses evoked by spinal stimulation were greater in SHR than WKY (p less than 0.01). Captopril reduced responses to electrical stimulation and this reduction was greater in the SHR (p less than 0.001). Bilateral nephrectomy reduced the vasoconstrictor responses to nerve stimulation in both strains of rat and abolished the effects of captopril. In an isolated perfused mesenteric artery WKY (p less than 0.05). Angiotensin II potentiated responses from both strains of rat, however the amplitude of the potentiation was greater in preparations from the SHR than those from WKY (p less than 0.002). Captopril (30 mg/kg by mouth) reduced blood pressure in conscious SHR over a 5-day dosing period. In WKY rats, no hypertensive action of captopril was observed. However, in another normotensive strain, the Alderley Park Wistar rat (APW), captopril lowered blood pressure. Plasma renin activity was not significantly different among these three strains of rat. The APW have previously been shown to be very sensitive to the adrenergic potentiating actions of angiotensin II. Captopril thus lowers blood pressure in SHR and APW, and both these strains are sensitive to the adrenergic potentiating actions of angiotensin II. It does not lower blood pressure in WKY, which is relatively insensitive to these actions of the octapeptide. Therefore, the hypotensive action of captopril in the rat may be due to its interference with the adrenergic potentiating effect of angiotensin II. PMID- 6292084 TI - Central effects of prostaglandin E2 on blood pressure and plasma renin activity in rats. Role of the sympathoadrenal system and vasopressin. AB - This study was designed to determine the roles of the sympathetic nervous system, adrenal medulla, and arginine vasopressin (AVP) in mediating pressor and plasma activity (PRA) responses to intraventricularly (ICV) administered prostaglandin E2 (PGE2) in conscious rats. The ICV PGE2 elevated blood pressure and caused increases in PRA, plasma AVP, and plasma norepinephrine and epinephrine. The pressor effect of ICV PGE2 was not influenced by pretreatment with captopril, but was attenuated by the AVP antagonist, d(CH2)5Tyr(Me)AVP, and by phenoxybenzamine, and was completely abolished by the combination of the AVP antagonist and phenoxybenzamine. The PRA response to ICV PGE2 was not affected by bilateral renal denervation or by phenoxybenzamine alone, but was attenuated by propranolol alone and was completely abolished by the combination of propranolol and phenoxybenzamine. Bilateral adrenomedullectomy did not affect the pressor response to ICV PGE2, whereas it attenuated the increase in PRA and completely abolished the increase in plasma epinephrine. These results suggest that the pressor effect of ICV PGE2 is the result of increased sympathetic nervous system activity and is dependent on the stimulation of alpha-adrenergic receptors and on AVP release. The pressor response to ICV PGE2 is accompanied by but not dependent on an increase in PRA. The renin-stimulating effect of centrally administered PGE2 is, at least in part, dependent on beta-adrenergic receptor stimulation by increased circulating catecholamines. PMID- 6292085 TI - Catecholamines in discrete kidney regions. Changes in salt-sensitive Dahl hypertensive rats. AB - Steady state levels of catecholamines (dopamine, norepinephrine, and epinephrine) were measured by the use of radioenzymatic techniques in discrete areas of the kidney (outer and inner cortex, outer and inner medulla) dissected by a "punch" technique from frozen kidney sections of salt-sensitive (DS) and salt-resistant (DR) Dahl rats fed a low or high salt diet. All three catecholamines were present in all areas of the kidney examined. There were gradients of concentrations of each catecholamine in different kidney areas. Renal medullary areas contained proportionally more dopamine than cortical areas. The proportion of epinephrine with respect to the total catecholamine content was relatively high in the inner medulla. Genetic factors and the amount of dietary salt influenced the catecholamine content in specific kidney areas, and these changes were different according to the area considered. DS rats when fed a high salt diet presented increased systolic blood pressure but no increased levels of dopamine in the inner medulla nor of norepinephrine in the outer medulla and outer cortex. Results suggest that either the uptake, release, storage, synthesis, or catabolism of kidney catecholamines is altered in Dahl salt-sensitive (DS) hypertensive rats and suggest specific roles for each catecholamine in discrete areas of the kidney. PMID- 6292086 TI - Renal alpha-adrenergic receptor abnormality in the spontaneously hypertensive rat. AB - Activation of renal alpha-adrenergic receptors induces vasoconstriction, proximal tubular reabsorption of sodium, and inhibition of renal release. Excesses of these effects are present in varying degrees in animal models of, and in patients with, "essential" hypertension. Since essential hypertension is genetically determined, we sought abnormalities of renal alpha-adrenergic receptors in the Okamoto-Aoki strain of spontaneously hypertensive rats (sr-SHR) and their stroke prone variant (sp-SHR). Total alpha-adrenergic receptor concentrations were determined by Scatchard analysis of [3H]dihydroergocryptine binding to a renal membrane fraction and were found to be increased (p less than 0.02) in male sr SHR at 4, 16, and 32 weeks of age and in female sr-SHR at 16 weeks of age as compared to age- and sex-matched Wistar-Kyoto controls. They were also increased in 9-week-old sp-SHR renal membranes (p less than 0.005). Further studies revealed that this increase in renal alpha-adrenergic receptors was due entirely to an increase in alpha 2-receptors as measured by [3H]yohimbine binding rather than to an increase in alpha 1-receptors as quantitated by [3H]prazosin binding. No difference in binding affinities of the various radioligands could be demonstrated between any of the hypertensive and normotensive groups of rats. Plasma norepinephrine levels were elevated (p less than 0.01) in the 4-, 9- and 16-week-old SHR, but not in the 32-week-old hypertensive rats. Thus, high renal alpha 2-adrenergic receptor number is coupled with a significant increase in plasma norepinephrine concentrations during the development of hypertension in SHR. By mediating an enhanced receptor-coupled response, such as increased proximal tubular sodium reabsorption, this abnormality of renal alpha-adrenergic receptors may contribute to some or all of the pathophysiologic derangements leading to hypertension in SHR. PMID- 6292087 TI - Hypertension and neurofibromatosis. Case report. AB - A 14-year-old girl with neurofibromatosis presented with severe hypertension. She was subsequently found to have a cerebellar glioblastoma multiforme and vascular lesions producing coarctation of the abdominal aorta and 50% and 95% stenosis of the left and right renal arteries respectively. No evidence of pheochromocytoma was found. After removal of the cerebellar tumor, marked amelioration of the hypertension suggested that the tumor had a major role in the pronounced elevation of her blood pressure. Patients who have both neurofibromatosis and hypertension should be carefully evaluated for these several potential lesions. PMID- 6292088 TI - Effect of propranolol and phenoxybenzamine on muscle strength and physical endurance, and on exercise-induced changes in muscle glycogen and blood lactic acid levels in rats. AB - Propranolol and phenoxybenzamine (PBZ) had no major effect on swimming-endurance performance and skeletal muscle pulling-strength in rats. Propranolol, like exercise itself diminished the resting skeletal muscle glycogen (SMG) content, but the drug did not lower the resting blood lactic acid (BLA) levels. Propranolol significantly antagonised the 25-min exercise-induced BLA elevation, indicating that lactacidaemia is possibly a beta-receptor response. However, propranolol did not exhibit an overall beneficial effect on swimming endurance. PBZ had a negligible effect on maximal swimming time (MST) and had no major effects on the SMG content and BLA levels. PMID- 6292089 TI - Ephedrine modifies hypothermia of clonidine and chlorpromazine. PMID- 6292090 TI - A study of neutralizing antibodies against polioviruses in vaccinated children. PMID- 6292091 TI - Oxygen dependence of human alveolar macrophage-mediated antibody-dependent cytotoxicity. AB - We studied the metabolic characteristics of the human alveolar macrophage mediated antibody-dependent cytotoxicity (ADCC) reaction, using an anti-D sensitized human erythrocyte target system. Metabolic experiments demonstrated a high resting rate of glucose metabolism in macrophages, but no oxidative metabolic burst was found to accompany the ADCC reaction. These findings were confirmed by oxygen consumption studies, showing a high resting rate of oxygen consumption by macrophages, but no change in the rate of oxygen consumption upon the addition of antibody-sensitized target cells. An anaerobic mechanism for ADCC was anticipated and investigated. Surprisingly, the macrophage-mediated ADCC reaction was found to be highly oxygen dependent. The macrophages of one chronic granulomatous disease patient were also studied and found to have a very low rate of oxidative metabolism in response to phagocytic stimuli. With oxygen present, these macrophages failed to produce significant ADCC, suggesting again that some oxidative mechanism was necessary in the macrophage-mediated ADCC reaction. Various oxygen radical scavengers were also studied. Glutathione inhibited ADCC significantly, and benzoic acid inhibited ADCC only slightly. All other scavengers had no significant inhibitory effect. Then, a known antioxidant and inhibitor of mixed-function oxidases, diethyldithiocarbamate, was found to produce a significant inhibition of the ADCC reaction. We believe this compound may be scavenging or inhibiting the production of some oxygen-dependent species important in the ADCC reaction. PMID- 6292092 TI - Use of monoclonal antibodies for analysis of antibody-dependent immunity to ocular herpes simplex virus type 1 infection. AB - Monoclonal antibodies specific for the five major glycoproteins of herpes simplex virus type 1 (HSV-1) were tested for their capacity to mediate immunity to ocular HSV-1 infection. The specificity of the immunoglobulin made by each monoclone was determined by immunoprecipitation of [14C]glucosamine-labeled polypeptides from detergent-solubilized HSV-1-infected cells. Of the five monoclonal antibodies studied, two immunoprecipitated glycoproteins gA/B, one immunoprecipitated glycoprotein gC, one immunoprecipitated glycoprotein gD, and one immunoprecipitated glycoprotein gE. All five were effective in passively transferring immunity to mice when they were given 4 to 24 h after HSV-1 infection on an abraded cornea. Four of the monoclonal antibodies were also evaluated for their capacity to neutralize HSV-1 and to promote complement mediated cell lysis and antibody-dependent cellular cytotoxicity. It was found that none of these in vitro assays correlated with the protective activity of the antibodies in vivo. In fact, one of the monoclonal antibodies was unreactive in all three immunological reactions, even though it was highly effective in promoting recovery from HSV-1 induced ocular disease in vivo. The results suggest that antibodies can interact in vivo with virus-specific glycoproteins gA/B, gC, gD, and gE to initiate recovery from HSV-1-induced ocular disease, and that the therapeutic effectiveness of a specific monoclonal antibody does not correlate with its immunological reactivity in vitro. PMID- 6292094 TI - Monoclonal antibodies to cytomegalovirus: rapid identification of clinical isolates and preliminary use in diagnosis of cytomegalovirus pneumonia. AB - Two monoclonal antibodies which react specifically with cells infected by cytomegalovirus (CMV) are described. One antibody, 6-E3, reacts with a 72,000 dalton protein that appears early in infection and remains localized in the cell nucleus. The other antibody, 6-C5, reacts with an 80,000-dalton protein that appears late in infection and remains localized in cytoplasmic inclusion bodies. Both monoclonal antibodies react with conventional laboratory strains of CMV and can be used in immunofluorescence assays to identify clinical isolates of CMV in culture. Preliminary tests on lung tissues from patients with CMV pneumonia show that only antibody 6-C5 detects CMV infection in primary clinical specimens. A comparison of culture, histological, and immunological methods demonstrates that the monoclonal antibodies possess sufficient specificity and sensitivity to warrant their continued development as immunodiagnostic tools for the detection of CMV infection in both tissue culture and tissues obtained directly from patients. PMID- 6292093 TI - Cytotoxic effector cells from infectious mononucleosis patients in the acute phase do not specifically kill Epstein-Barr virus genome-carrying lymphoid cell lines. AB - We describe a study in which we investigated the cytotoxic activities of thymusderived (T) lymphocytes and natural killer cells against Epstein-Barr virus (EBV) genome-carrying lymphoid cell lines. Purified subpopulations of lymphocytes from eight patients with infectious mononucleosis and six healthy normal EBV seropositive donors were tested. Enriched T-cells were obtained by passing purified whole blood lymphocyte preparations through human immunoglobulin-anti immunoglobulin-coated glass bead columns. The cytolytic activity of effector cells was determined by the ability of these cells to lyse human target cells that were internally labeled with (51)Cr. These targets included cells from both EBV genome-carrying and EBV genome-negative lymphoid lines derived from malignant tumors, as well as from lymphocytes transformed in vitro by EBV, and were chosen to represent a wide spectrum of EBV-associated membrane antigens. We found that cytotoxic T-cells from patients with infectious mononucleosis showed no EBV related specific cell killing per se, although a trend for increased killing of cell lines derived from spontaneous in vivo growing tumors, EBV genome carrying or not, was noted; however, this trend was not observed with cell lines derived from cord blood lymphocytes after EBV infection in vitro. In addition, our data suggest that natural killer cells may play an important role in controlling EBV infection in patients with infectious mononucleosis in the acute phase of the disease, particularly since T-cells (obtained after removal on immunoglobulin anti-immunoglobulin columns of natural killer cells presumably bearing Fc receptors) were less efficient killers than whole blood lymphocytes; furthermore, lysis by whole blood lymphocytes was also greatest against cell lines derived from malignant tumors (as opposed to in vitro EBV-transformed cord blood lymphoid lines), irrespective of whether these targets were EBV genome positive or negative. PMID- 6292095 TI - Characterization of canine neutrophil granules. AB - The purpose of this study was to isolate distinct populations of canine neutrophil granules and to compare them with neutrophil granules from other species. Size, shape, density, and content of canine neutrophil granules were determined. Neutrophils obtained by Ficoll-Hypaque sedimentation were homogenized, and granule populations were separated by isopycnic centrifugation on a linear sucrose gradient (rho, 1.14 to 1.22 g/ml). The most dense granule population (rho, 1.197 g/ml) contained all of the myeloperoxidase, beta glucuronidase, and elastase, more than half of the acid beta-glycerophosphatase, and most of the lysozyme. The population with intermediate density (rho, 1.179 g/ml) contained lactoferrin, vitamin B12-binding protein, and the remainder of the acid beta-glycerophosphatase and lysozyme. The least dense granule population did not contain a major peak of any of the enzymes or binding proteins tested but was distinguished by density and morphology. The size and shape of the granules were determined from scanning electron micrographs and assessment of shape was aided by transmission electron micrographs. By these methods three populations of canine neutrophil granules were characterized and named: myeloperoxidase granules, vitamin B12-binding protein granules, and low-density granules. PMID- 6292096 TI - Fate of vaccines of Propionibacterium acnes after phagocytosis by murine macrophages. AB - Stationary-phase (48 h) cells of Propionibacterium acnes VPI 0009, a potent stimulator of the reticuloendothelial system, persist unchanged within phagocytes for at least 24 h after ingestion. In contrast, exponential-phase (12 h) cells of the same strain (which do not induce splenomegaly) are extensively degraded within 5 h of phagocytosis. Suspensions of P. granulosum VPI 6500, which fails to induce splenomegaly in mice, also show considerable degradation after phagocytosis. Stationary-phase cells of strain VPI 0009 treated with sodium metaperiodate or with trichloroacetic acid, although without ability to induce splenomegaly, resist destruction almost as well as untreated vaccines. However, bacteria inactivated by acetic anhydride show about 50% breakdown in 24 h. PMID- 6292097 TI - Mitogenic activity in human embryonic fibroblasts early after infection by human cytomegalovirus. AB - We have characterized the nonspecific lymphocyte stimulation by extracts of human cytomegalovirus-infected human embryonic fibroblasts. Cell extracts prepared at 5 h postinfection (early extract) and 72 h postinfection (late extract) were both highly mitogenic in lymphocyte preparations from adult blood, cord blood, and rabbit blood. Maximum stimulation of the lymphocytes was observed on day 3 after the addition of early or late extract under optimal conditions. Early extract stimulated both the E-rosetting and the EAC-rosetting subpopulations of human lymphocytes. The mitogenic activity appeared before 5 h postinfection and was fairly stable at 30 degrees C for 5 h. PMID- 6292098 TI - Clostridium perfringens type A enterotoxin: characterization of the amino terminal region. AB - The amino-terminal region of the enterotoxin of Clostridium perfringens was investigated by automated sequence analysis. The primary structure results revealed that the enterotoxin is composed of a single polypeptide amino acid sequence. Computer comparison of a 20-residue sequence with a sequence library of reported proteins revealed no significant chemical similarities, indicating that the enterotoxin represents a unique polypeptide primary structure. PMID- 6292099 TI - Retrovirus antigens in brains of mice with scrapie- and murine leukemia virus induced spongiform encephalopathy. AB - Wild mouse ecotropic virus-induced spongiform encephalomyelopathy pathologically similar to scrapie was associated with the expression of retrovirus antigens in mouse brains. However, scrapie-infected mice with spongiform encephalopathy showed no increased expression of retrovirus antigens in brain. Thus, the pathogenesis of the scrapie spongiform lesion does not appear to involve activation of endogenous retrovirus. PMID- 6292100 TI - Different sugar compositions of lipopolysaccharides isolated from phase I and pure phase II cells of Coxiella burnetii. AB - The compositions of lipopolysaccharides I and II, isolated from Coxiella burnetii phase I and pure phase II cells, respectively, showed drastic and clear-cut differences. Lipopolysaccharide II exhibited a comparatively simple composition. It contained, in addition to glucosamine (from lipid A) and a 2-keto-3 deoxyoctulosonic acid-related, thiobarbituric acid-positive substance, only two aldoses in high amounts, namely D-mannose and D-glycero-D-mannoheptose. Lipopolysaccharide I contained the same components, but it also contained at least eight other constituents including neutral sugars, e.g., an unusual branched sugar (3-C-methyl-6-deoxy-hexose) and unusual aminohexoses. Some components were characterized by a very high retention time in gas-liquid chromatography. The similarity of the phase variation in C. burnetii to the well known smooth-to-rough mutation found in Enterobacteriaceae is discussed. Cells of phase I correspond to enterobacterial smooth forms, whereas cells of phase II might be compared to the rough forms. PMID- 6292101 TI - Occurrence and frequency of coronavirus infections in humans as determined by enzyme-linked immunosorbent assay. AB - The occurrence of human coronavirus (HCV) infections was analyzed by using sequential sera taken between 1976 and 1981 from adults working in the London area. Antibody rises to HCV 229E and HCV OC43 group viruses were measured in serum samples from these subjects by enzyme-linked immunosorbent assay. HCV infections were found throughout the year, although most occurred during two periods, from June through September and from December through February. There were no marked seasonal differences in either the range of antibody rises obtained or in the HCV groups to which these antibody rises were directed. However, there were more HCV antibody rises during the summer than in the winter. The antibody duration varied considerably, but had a mean of 3.5 months. Finally, the frequency of HCV infection per person was calculated to be 1 per 7.8 months. PMID- 6292102 TI - Neutralizing antibody response of rabbits and goats to caprine arthritis encephalitis virus. AB - Rabbits were immunized with purified caprine arthritis-encephalitis virus and examined for neutralizing activity. Analysis of virus-antiserum interaction at 37 degrees C demonstrated little loss of viral infectivity after incubation with heat-inactivated rabbit antiserum for 60 min. However, sensitization of virus (as assessed by the addition of complement) occurred almost immediately and was 95% complete after 10 min. The complement-dependent neutralizing activity was associated with the immunoglobulin G fraction of rabbit antiserum. Addition of goat anti-rabbit immunoglobulin G to the immune rabbit serum-caprine arthritis encephalitis virus mixture also resulted in neutralization of infectivity when unbound antibody was removed before addition of the anti-immunoglobulin. Serum from most caprine arthritis-encephalitis virus-infected goats contains antibody activity to the core protein p28, as demonstrated by immunodiffusion and enzyme linked immunosorbent assay. However, attempts to demonstrate neutralizing activity in the serum of goats up to 1.5 years post-inoculation or in serum of hyperimmunized goats were unsuccessful when the sera were examined alone or in combination with complement or rabbit anti-goat immunoglobulin or both. PMID- 6292104 TI - Failure to detect hemagglutination-inhibiting antibodies with intact avian influenza virions. AB - Avian influenza viruses replicate in a variety of mammals and birds, yet hemagglutination inhibition tests show that postinfection sera from these animals (e.g., ferrets and ducks) have insignificant levels of antibodies (Hinshaw et al., Infect. Immun. 34:354-361, 1981). This suggested that avian influenza viruses, in contrast to mammalian viruses, may not induce a significant humoral response. Studies reported here indicate that avian influenza viruses do induce high levels of antibodies in ferrets, ducks, and mice and produce long-lived memory for cytotoxic T-cells in mice. The failure to detect hemagglutination inhibiting antibodies to avian viruses was explained by the finding that antibodies to avian influenza viruses were not detectable in hemagglutination inhibition tests with intact virus yet were readily demonstrable when hemagglutinin subunits were used. In addition, these sera contained high levels of neutralizing antibodies to the avian virus. These findings suggest that the hemagglutinins of avian and mammalian influenza viruses may differ in their accessibility to antibodies or the biological consequence of antibody attachment or both. The practical consequence of these studies is that hemagglutination inhibition tests with intact avian viruses fail to detect antibody and do not correlate with virus neutralization. The avian virus used in these studies, A/Mallard/NY/6870/78 (H2N2), replicated and caused mortality in BALB/c mice, emphasizing that the host range and virulence of avian viruses extends to mammals. The above findings suggest that avian viruses could infect mammals in nature, yet seroepidemiological studies with conventional hemagglutination inhibition tests could give misleading results. PMID- 6292103 TI - Damage to Aspergillus fumigatus and Rhizopus oryzae hyphae by oxidative and nonoxidative microbicidal products of human neutrophils in vitro. AB - Our previous studies established that human neutrophils could damage and probably kill hyphae of Aspergillus fumigatus and Rhizopus oryzae in vitro, primarily by oxygen-dependent mechanisms active at the cell surface. These studies were extended, again quantitating hyphal damage by reduction in uptake of (14)C labeled uracil or glutamine. Neither A. fumigatus nor R. oryzae hyphae were damaged by neutrophils from patients with chronic granulomatous disease, confirming the importance of oxidative mechanisms in damage to hyphae. In contrast, neutrophils from one patient with hereditary myeloperoxidase deficiency damaged R. oryzae but not A. fumigatus hyphae. Cell-free, in vitro systems were then used to help determine the relative importance of several potentially fungicidal products of neutrophils. Both A. fumigatus and R. oryzae hyphae were damaged by the myeloperoxidase-hydrogen peroxide-halide system either with reagent hydrogen peroxide or enzymatic systems for generating hydrogen peroxide (glucose oxidase with glucose, or xanthine oxidase with either hypoxanthine or acetaldehyde). Iodide with or without chloride supported the reaction, but damage was less with chloride alone as the halide cofactor. Hydrogen peroxide alone damaged hyphae only in concentrations >/=1 mM, but 0.01 mM hypochlorous acid, a potential product of the myeloperoxidase system, significantly damaged R. oryzae hyphae (a 1 mM concentration was required for significant damage to A. fumigatus hyphae). Damage to hyphae by the myeloperoxidase system was inhibited by azide, cyanide, catalase, histidine, and tryptophan, but not by superoxide dismutase, dimethyl sulfoxide, or mannitol. Photoactivation of the dye rose bengal resulted in hyphal damage which was inhibited by histidine, tryptophan, and 1,4 diazobicyclo(2,2,2)octane. Lysates of neutrophils or separated neutrophil granules did not affect A. fumigatus hyphae, but did damage R. oryzae hyphae. Similarly, three preparations of cationic proteins purified from human neutrophil granules were more active in damaging R. oryzae than A. fumigatus hyphae. This damage, as with the separated granules and whole cell lysates, was inhibited by the polyanion heparin. Damage to R. oryzae hyphae by neutrophil cationic proteins was enhanced by activity of the complete myeloperoxidase system or by hydrogen peroxide alone in subinhibitory concentrations. These data support the importance of oxidative products in general and the myeloperoxidase system in particular in damage to hyphae by neutrophils. Cationic proteins may also contribute significantly to neutrophil-mediated damage to R. oryzae hyphae. PMID- 6292106 TI - Antibody-dependent cell-mediated cytotoxicity against varicella-zoster virus infected targets. AB - Antibody-dependent cell-mediated cytotoxicity (ADCC) against cryopreserved varicella-zoster virus-infected human foreskin fibroblasts was detected in a 51Cr release assay. Target cells, samples of seropositive or seronegative sera, and mononuclear cells obtained by Ficoll-Hypaque centrifugation of human peripheral blood were added to microtiter plate wells and allowed to incubate at 37 degrees C for 4 h. Fibroblasts infected for 48 to 96 h were susceptible to ADCC. Effector cells from seropositive and seronegative normal children were equally active in the assay. Antibody titers were determined by testing serial dilutions of sera in the ADCC assay. Zoster immune globulin had a titer of 204,800. Sera from 40 naturally seropositive individuals were compared by assays for ADCC and fluorescent antibody to membrane antigen. All sera that were negative by fluorescent antibody to membrane antigen (less than 2) were also negative by ADCC (less than 20). All sera that were positive by fluorescent antibody to membrane antigen were also positive by ADCC, but titers of individual sera were frequently 5 to 20 times higher in the ADCC assay. PMID- 6292105 TI - Effect of pyridines on phenotypic properties of Bordetella pertussis. AB - Several conditions of growth of Bordetella pertussis cause a reversible phenotypic alteration in properties termed modulation. Growth in medium containing nicotinic acid induces normal (X-mode) cells to change to modulated (C mode) cells. We examined several pyridines and compounds resembling pyridines for their ability to affect modulation, using envelope protein patterns and serological reactivity as indicators of modulation. We found that 6 chloronicotinic acid and quinaldic acid were more effective modulating stimuli than was nicotinic acid on a molar basis. Both 2-chloronicotinamide and isoniazid interfered with nicotinic acid-induced modulation, and can be called antimodulators. Picolinic acid inhibited growth. PMID- 6292107 TI - Purification of two Clostridium perfringens enterotoxin-like proteins and their effects on membrane permeability in primary cultures of adult rat hepatocytes. AB - We isolated two proteins, ET-1 and ET-2, from the sporangial extracts of Clostridium perfringens type A. Both proteins had some properties in common with the well-known C. perfringens enterotoxin. ET-1 and ET-2 behaved as single and distinct entities in anion exchange chromatography and disk gel electrophoresis. ET-2 was the more anionic protein since it eluted more slowly from the anion exchange column and migrated faster toward the anode in polyacrylamide disk gel electrophoresis (pH 8.5, native gels). Additionally, in this electrophoretic system ET-2 was not distinguishable from the enterotoxin. The amino acid compositions of ET-1 and ET-2 were similar but differed in a few amino acid residues. The values for both proteins were also similar to the published reports of others for the enterotoxin. Both ET-1 and ET-2 showed lines of identity in agar gel double immunodiffusion against anti-enterotoxin antiserum. Both ET-1 and ET-2 were toxic for rat hepatocytes in primary monolayer culture as determined by accelerated exodus of L-[14C]glucose from preloaded cells and by the rapid uptake of 45Ca2+ after exposure to the proteins. In this regard, ET-1 and ET-2 appeared to be identical in mechanism of action to what has been regarded in the literature as "the" C. perfringens enterotoxin. Interestingly, ET-2 was 3 to 10 times more toxic on a weight basis than ET-1 was. PMID- 6292108 TI - Hydrophobic interactions and the adherence of Streptococcus sanguis to hydroxylapatite. AB - Streptococcus sanguis demonstrated a high affinity for hydrocarbon solvents. When aqueous suspensions of the organism were mixed with either hexadecane or toluene, the cells tended to bind to the nonaqueous solvent. Increases in temperature resulted in a greater affinity of cells for hexadecane. Interaction between the cells and hexadecane was also enhanced by dilute aqueous sodium chloride and by low pH (pH less than 5). The results suggest that the cell surface of S. sanguis has hydrophobic properties. Isolated cell walls also tended to partition into the nonaqueous solvent. Amino acid analyses of the walls revealed the presence of several amino acids which possess hydrophobic side chains. It is likely that the hydrophobic amino acids associated with the cell wall contribute to the hydrophobicity of intact S. sanguis. When the adherence of S. sanguis to saliva coated hydroxylapatite was measured, it was found that hydrophobic bond disrupting agents, such as the Li+ cation, the SCN- anion, and sodium dodecyl sulfate, were capable of inhibiting the cell-hydroxylapatite union. In addition, it was observed that both urea and tetramethylurea were inhibitors of the adherence, although the latter reagent was the superior inhibitor. The results suggest that the adherence of S. sanguis to saliva-coated smooth surfaces is at least partially dependent on the formation of hydrophobic bonds between the cell and adsorbed salivary proteins. Hydrophobic bonding may contribute to cooperative interactions involving S. sanguis and saliva-coated hydroxylapatite (Nesbitt et al., Infect. Immun. 35:157-165, 1982). PMID- 6292109 TI - Pathogenicity of avian reoviruses: examination of six isolates and a vaccine strain. AB - Six avian reovirus isolates and a vaccine reovirus strain were compared for invasiveness, virulence, and pathological characteristics upon infection of day old specific-pathogen-free chicks by the footpad, subcutaneous, and oral routes of inoculation. No significant differences were noted regarding the ability of individual isolates to infect target tissues. However, virulence (measured as the 50% lethal dose) among the isolates varied markedly from 2 x 10(5) to less than 10 PFU per chick for the most virulent isolate; between the parental wild-type virus and the derivative vaccine virus strain, a million-fold (10(6)) difference in virulence was demonstrated. All strains revealed, with considerable variation, arthrogenic potential. PMID- 6292110 TI - Pathogenesis of rotavirus infection in mice. AB - Three parameters of rotavirus infection, i.e., clinical disease, viral antigen in infected intestines, and infectious virus in feces, were assessed in infant mice nursed by mothers with or without preexisting rotavirus antibody. Diarrhea was the only consistent sign of clinical disease, and its course followed that of infection by about 1 day. Infected intestinal epithelial cells, except crypt cells, were observed by immunofluorescence microscopy in the duodenum, jejunum, ileum, and colon. Infection progressed in a proximal-to-distal direction with time. Viral antigen appeared in intestinal tissue later, was present in lower amounts, and disappeared sooner from infants nursed by mothers with preexisting rotavirus antibody, indicating that protection was passively transferred to these infants although the course of clinical disease was not changed. PMID- 6292111 TI - Propagation and primary isolation of papovavirus JC in epithelial cells derived from human urine. AB - Human papovavirus JC, previously passaged in amnion cells or in primary human fetal glial cells, replicated efficiently in urine-derived epithelial cells. Primary isolation of the virus from brain extracts was possible in urine-derived cells, but these cells were not as sensitive as primary human fetal glial cells for this purpose. Primary isolations of human papovavirus JC from urine sediments of renal transplant patients were made in urine-derived cells. PMID- 6292112 TI - Three human rotavirus serotypes demonstrated by plaque neutralization of isolated strains. AB - Human rotaviruses were isolated directly from stool specimens of gastroenteritis patients in MA-104 cells in the presence of trypsin. For the plaque assay of the isolated strains, the optimal composition of overlay medium was determined. The antigenicity of the isolated strains was investigated by a plaque neutralization method, using antisera prepared against six strains having different electropherotypes of viral RNA, and three different neutralization serotypes were demonstrated. PMID- 6292113 TI - Herpes simplex virus infection of human T-cell subpopulations. AB - The ability of herpes simplex virus type 1 to productively infect human T-cell subpopulations was examined. Unstimulated helper/inducer (T4+) and cytotoxic/suppressor (T8+) lymphocytes limited herpes simplex virus replication as effectively as unseparated peripheral blood T cells (T3+). Phytohemagglutinin stimulation before infection resulted in equivalently productive herpes simplex virus infections in the three cell fractions. PMID- 6292115 TI - The interaction of non-transforming Epstein-Barr virus (EBV) with cell-associated EBV DNA in superinfected lymphoblastoid cell lines. AB - Two human lymphoblastoid cell lines were established by the transformation of human cord-blood lymphocytes with transforming Epstein-Barr virus (EBV). One cell line (HLB-R1) was established with EBV obtained after the superinfection of Raji cells with HR-1 EBV and the other (HLB-Bl) was established from B95-8 EBV infected human cord-blood lymphocytes. Both the HLB-R1 and HLB-B1 lines were susceptible to superinfection with HR-1 EBV. We found that EBV DNA was replicated in the superinfected cell lines and that transforming EBV was produced in both the HLB-B1 and HLB-R1 cells. The average titer of transforming EBV obtained in the HR-1 EBV superinfected HLB-B1 and HLB-R1 cell lines was 10(4) transforming units (TU)/ml, whereas the average titers of transforming EBV obtained by the superinfection of Raji cells was 10(1) TU/ml. Epstein-Barr virus capable of inducing early antigen (EA) in superinfected Raji cells (lytic virus) was not detected in any transforming virus preparation. Restriction enzyme digestion patterns of virus DNA isolated from HR-1 and B95-8 cells, as well as from superinfected cells, were compared. The EBV DNA that was replicated in the superinfected HLB-R1 and HLB-B1 cell lines showed a more complex pattern. Our data suggest that recombination between input HR-1 EBV DNA and latent cell associated EBV DNA occurs. Presumably this recombination results in a change in the biological properties of the newly synthesized virus. PMID- 6292114 TI - Asbestos-induced alteration of human peripheral blood monocyte activity. AB - Incubation of chrysotile and anthophyllite asbestos fibers with normal human peripheral blood monocytes resulted in significant suppression of monocyte metabolic activity as measured by chemiluminescence. Both fiber types were cytotoxic to monocytes and depressed monocyte phagocytosis of latex beads. We conclude that asbestos-induced monocyte cytotoxicity could result in release of lysosomal enzymes and/or degradation products which contribute to fibrosis in asbestosis. The depression of phagocytosis and microbicidal function may contribute to the increased incidence of carcinogenesis observed in asbestosis. PMID- 6292116 TI - Epstein-Barr virus strain-specific differences in transformed cell lines demonstrated in growth characteristics, induction of viral antigens and ADCC susceptibility. PMID- 6292117 TI - Detection of antibodies against the cellular protein p53 in sera from patients with breast cancer. AB - Antibodies reacting with the host protein p53 were found in the sera of patients with primary or secondary carcinoma of the breast. Fourteen out of the 155 sera from breast cancer patients tested were positive for anti-p53 antibodies (9%) and no positives were detected among 164 control sera from normal women tested. The locations of the first metastasis in patients with positive sera were unusual, with more lung metastases and fewer bone metastases than expected. The detection of anti-p53 antibodies indicates that p53 is altered in amount, type or presentation in breast tumors so that it becomes immunogenic. PMID- 6292118 TI - Induction of avidin in chickens infected with the acute leukemia virus OK 10. AB - The presence of avidin, a progesterone-dependent oviductal glycoprotein, was studied in viral tumors. Newborn chickens were infected with the acute leukemia virus OK 10, and the first tumors occurred within 2-3 weeks. Avidin was assayed using a [14C]biotin-binding method and radioimmunoassay. In the control chickens, avidin concentrations were less than 0.3 microgram/g in the plasma and less than 1.5 microgram/g in various tissues including the immature oviduct. In the OK 10 virus-infected chickens, no significant induction was observed during the acute infection or any time thereafter if no tumors were seen. In chickens that developed tumors, avidin concentrations were significantly increased in tumorous tissue only located in the mesenterium and occasionally in the oviduct. In tumors occurring elsewhere (kidney, ovary, muscle, testis, liver, colon) avidin concentrations were not elevated. Tumor-associated avidin had extraordinary biotin-binding capacity after treatment at +90 degrees C similar to the progesterone-dependent avidin, whereas antibody-binding properties suggested that tumor-associated avidin may have a somewhat altered antigenic structure. PMID- 6292119 TI - Epstein-barr virus induction by a serum factor: IV. Ubiquitous occurrence of the factor within vertebrates and its interaction with defined lymphoid cell lines. AB - We have recently reported the existence of a serum factor that induces Epstein Barr virus antigens in Raji cells (Bauer et al., 1982b, c). Here we demonstrate the ubiquitous presence of this factor in sera of vertebrates of all five classes. Furthermore, we tested 48 EBV-genome-positive lymphoid cell lines of different origins for responsiveness to the serum factor. Only cells from lines of two groups, i.e. Burkitt's lymphoma and EBV-transformed marmoset lines, responded to the factor, whereas human lymphoblastoid cell lines were not induced by the serum factor. PMID- 6292120 TI - Pyrimidine biosynthetic enzymes in Babesia hylomysci. PMID- 6292121 TI - Effect of oxygen and misonidazole on radiation damage in biologically active DNA dissolved in a bacterial extract. Influence of cytochrome c. PMID- 6292122 TI - Epidermal growth factor: mechanisms of action. PMID- 6292123 TI - Enucleation and the appearance of second primary tumors in cats bearing virally induced intraocular tumors. AB - The effect of enucleation of an eye containing a malignant intraocular neoplasm on the occurrence of secondary tumors was studied in cats with tumors, mainly melanomas, induced by Gardner feline fibrosarcoma virus. Enucleation of eyes containing progressively growing tumors was followed by a sharp increase in the frequency of secondary tumors. Secondary tumors were detected in 13 of 14 (92.8%) cats subjected to enucleation but in only seven of 21 (33.3%) untreated cats. The data suggest that the secondary tumors were not metastases but rather second primary tumors induced by local transformation of fibrosarcoma virus shed from the intraocular neoplasms. The increased incidence of these second primary tumors in cats subjected to enucleation was associated with depressed antibody titers to a tumor-specific transplantation antigen, the feline oncornavirus-associated cell membrane antigen. PMID- 6292124 TI - Ultrasonic diagnostic system for interactive interrogation of adult brain through intact skull. AB - An integrated ultrasonic system has been developed employing static gray scale imaging, digital image processing, and analyses of quantitative ultrasonic backscatter for interactive interrogation of brain through the adult human skull. Operating at 750 kHz, to avoid severe spatial and temporal pulse distortion which accompany ultrasonic transkull transmission at higher frequencies, storage and image processing of successive images taken at selected gain settings in the same image plane allow the reconstruction of a composite cross-section of the brain and skull and enable visualization of internal brain structure. Analyses of digitized backscatter data obtained with the visualization transducer over selectable interrogation paths referenced to the cross-sectional image permit quantitative classification of tissue type independent of qualitative visual image analysis, such that enhanced diagnostic potential is provided. A previous paper described in detail the modifications of the commercial ultrasound diagnostic unit which serves as the core of this system. The present paper presents the rationale for the integrated system design approach, as well as a description of graphics display and tissue classification features which have been incorporated into the instrumentation configuration and are deemed necessary for successful transkull ultrasonic imaging and diagnosis. PMID- 6292125 TI - Classification of hepatitis A virus as enterovirus type 72 and of hepatitis B virus as hepadnavirus type 1. PMID- 6292126 TI - Biophysical and biochemical characterization of hepatitis A virus. AB - Biophysical and biochemical analysis of hepatitis A virus has shown it to be a 27 to 32-nm icosahedral particle with 32 capsomers. The mature virion has a buoyant density of 1.33-1.34 g/cm3, a sedimentation coefficient of 156-160S, and is composed of four polypeptides with molecular weights of 30,000-33,000 (VP1), 24,000-27,000 (VP2), 21,000-23,000 (VP3), and 7,000-14,000 (VP4). The genome of hepatitis A virus consists of a single piece of single-stranded RNA which sediments at 32-35S and has a buoyant density of 1.64 g/cm3. The molecular weight of RNA is 2.25 x 10(6) when measured under nondenaturing conditions and 2.8 x 10(6) when measured under fully denaturing conditions. The genome contains a 40 80 nucleotide sequence of polyadenylic and is capable of infecting cell cultures. These findings, together with the observation that the virion is stable at pH 3.0 and resistant to ether and a temperature of 60 degrees for 1 h, indicate that hepatitis A virus should now be classified as an Enterovirus within the family Picornaviridae. PMID- 6292127 TI - Partial characterization of temperature-sensitive mutants of foot-and-mouth disease virus, O1 caseros strain. AB - The preliminary characterization of four temperature-sensitive (ts) mutants of the O1 Caseros strain of foot-and-mouth disease virus is described. Two mutants, ts 6 and ts 40, showed a very low RNA synthesis rate at nonpermissive temperature and were classified phenotypically as RNA(-). Shift-up experiments demonstrated an incapacity to organize the RNA replicative process at nonpermissive temperature. Another mutant (ts 139) behaved phenotypically as RNA(+) and its virions were more thermolabile than the wild type virus, so the defect in this mutant is likely to be in one of its structural proteins. Finally, mutant ts 5 was phenotypically RNA(+); its leak production at nonpermissive temperature was high, and the shift-up experiment showed a defect that was expressed either late in the replicative cycle or continuously. PMID- 6292128 TI - In vivo and in vitro models of demyelinating diseases. V. Comparison of the assembly of mouse hepatitis virus, strain JHM, in two murine cell lines. AB - The developmental sequence of a neurotropic strain (JHM) of mouse hepatitis virus was examined by transmission electron microscopy and immunocytology. The nucleoprotein core of this coronavirus, which contains RNA of positive polarity and is helical in configuration, becomes incorporated into enveloped particles in the same manner as the nucleocapsids of the orthomyxo- and paramyxoviruses. However, JHM virus is assembled intracellularly by budding at surfaces of smooth membranous vacuoles. A comparison of JHM virus replication in L2 and 17Cl-1 cell lines revealed that L2 cells undergo more rapid cytopathology and cease virus production much sooner than 17Cl-l cells. In L2 cells the accumulation of core material appears to continue after the abrupt cessation of virus assembly. This is evident by the massive cytoplasmic accumulation of structure resembling nucleocapsids, which react with hybridoma antibody to the nucleocapsid antigen as demonstrated by the immunoperoxidase procedure. The current findings are consistent with our previously published demonstration, using cells of neural and other deviation, of the fundamental role of the host cell type in regulating the replication and expression of coronaviruses. PMID- 6292129 TI - Microfilaments associated with Paget's disease of bone: comparison with nucleocapsids of measles virus and respiratory syncytial virus. AB - The structural characteristics and intracellular disposition of microtubules present in osteoclasts in Paget's disease of bone were compared with those of nucleocapsids in cells infected with measles virus and respiratory syncytial virus (RSV). The distribution of microtubules in pagetic osteoclasts closely paralleled that of measles nucleocapsids in monkey kidney cells, but the dimensions of the pagetic microtubules were significantly different from those of measles virus. In contrast, pagetic microtubules were not distinguishable in dimensions from RSV nucleocapsids but were dissimilar in their conformation and location. The observations support the hypothesis tht Paget's disease of bone is a slow virus disease and suggest that the virus is a pneumovirus (RSV) rather than a morbillivirus (measles). PMID- 6292130 TI - Kinetics of human antibody responses to primary genital herpes simplex virus infection. AB - 5 patients with primary genital herpes simplex virus (HSV) infection were studied prospectively for the production of serum antibody reactivity as measured by the virus micro-neutralization test (NT), micro-solid phase radioimmunometric assay (micro-SPRIA), and antibody-dependent cellular-cytotoxic (ADCC) test. ADCC antibody was detected before reactivity measured by either NT or micro-SPRIA in 2 of the patients. A 3rd patient failed to produce neutralizing antibody, whereas specific activity was detected by the other two methods. In 2 other patients a good correlation was observed for development of antibody by all three methods. There were several discordant results in the kinetics of production of antibody detected by these assays, suggesting that not all individuals produce similar types of antiviral antibody. PMID- 6292131 TI - Accumulation of poliovirus proteins in the host cell nucleus. AB - Poliovirus (type 1, Mahoney) proteins were transported into the nuclei of HEp-2 cells, as demonstrated by means of electron microscopic autoradiography. Quantitative determinations of the proteins by electrophoresis showed that the relative amounts of precursor proteins (1a, 3b, 1b), of some of the intermediate polypeptides (3a, 3c, 5b, 2), and of one end product (VP1) were higher in the nucleus than in the cytoplasm. Other proteins (VPO, VP2, X, 4, 6a) were found in the same relative amounts in the nucleus as in the cytoplasm, whereas two proteins (VP3 and 6b) were excluded from the nucleus. The findings are discussed in view of the recently demonstrated inhibitory activity exerted by cytoplasmic extracts of poliovirus-infected cells on transcription in isolated nuclei of uninfected cells. PMID- 6292132 TI - A mouse model of transmural myocardial necrosis due to coxsackievirus B4: observations over 12 months. AB - The course of coxsackievirus B4 necrotizing myocarditis was studied over a 12 month period in 712 ICR Swiss mice inoculated at less than 48 h of age. Affected animals were sacrificed at intervals until 1 year. Microscopically, focal myocardial necrosis, which was often transmural in extent with mixed inflammatory exudate, and subsequent fibrous replacement were induced in 75% of the examined animals. The left ventricle (63 of 69 subjects, 91.3%), interventricular septum (39 of 69 subjects, 56.5%) and right ventricle (26 of 69 subjects, 37.7%) were most frequently involved. Thinning of the ventricular wall and grossly apparent localized ventricular bulges (aneurysms) were seen in 22 subjects (the left ventricle 16 times, the right ventricle 5 times, and the interventricular septum 5 times). Coronary arteries were normal in all instances. PMID- 6292133 TI - Genital herpes simplex infections in Israel: 1973 throughout 1980. AB - Ninety-eight cases of genital herpes simplex virus infection (HSV-2) were confirmed by laboratory examinations during the years 1973-80. Until 1975 there were less than five cases per year, but from 1976 the number of cases gradually increased, reaching 24 in 1980. The age distribution is typical of a sexually transmitted disease; and since the peak of susceptibility is at childbearing age, an increase in neonatal HSV infection could be expected. In 1980, laboratory testing confirmed neonatal HSV infection in four newborns. The need to monitor pregnancies in women at high risk for HSV-2 infection is stressed. PMID- 6292134 TI - [Generalized eruptive histiocytoma]. AB - A 52-year-old male patient with generalized eruptive histiocytoma persisting for about 20 years is described. The clinical picture is characterized by red brownish, round to oval nodules histologically consisting mainly of histiocytes. The ultrastructure of these skin lesions shows histiocytic cells at various stages of differentiation with different degree of lipid storage. The generalized eruptive histiocytoma are separated from multicentric reticulohistiocytosis. The significant differences refer to the clinical picture, in particular the organ manifestations associated with the skin lesions. The histological and ultrastructural features of the generalized eruptive histiocytoma differ from this differential diagnosis for lack of giant cells and of interdigitations between the neighbouring cells. The relation between generalized eruptive histiocytoma and the multinodular reticulohistiocytosis is discussed. PMID- 6292135 TI - Hemoglobin Abidjan [alpha 51 (CE 9) Gly replaced by Asp] in a Japanese. PMID- 6292136 TI - Hb J Daloa (beta 57 (E1) Asn replaced by Asp): a new variant found in Ivory Coast. PMID- 6292137 TI - Dietary recommendations for diabetics for the 1980s--a policy statement by the British Diabetic Association: prepared by the Nutrition Sub-Committee of the British Diabetic Association's Medical Advisory Committee. PMID- 6292138 TI - The interaction of wheat bran and oral iron supplements in vivo. PMID- 6292139 TI - Cytochemical demonstration of amine-storing vacuoles and lysosomes in the chicken thrombocytes. AB - A combined electron microscopic and cytochemical study of the thrombocytes of the chicken has clearly identified the amine-storing organelles and lysosomes. A chromaffin positive-reaction product was observed on the inner surface and the granules of the large electron-lucent vacuoles. No acid phosphatase activity was localized in these amine-storing vacuoles. However, the acid phosphatase activity was observed in the small vesicles, the primary lysosomes, and in the large electron dense inclusions with myelin which may be secondary lysosomes. The results of this study suggest that the large empty vacuoles, with one or two very dense osmiophilic peripherally-situated granules, in the chicken thrombocytes are comparable to the vesicles with electron dense materials called "dense bodies" in mammalian thrombocytes. PMID- 6292140 TI - [Granular cell tumors of the larynx]. AB - Based on four observations of our own and a survey of the 151 cases described to date, the clinical and morphologic results of granular cell tumours of the larynx are presented. These tumours, which were considered as being of myogenic origin ("myoblastic myoma", immature rhabdomyoma", "granular cell myoblastoma" etc.) are probably of neurogenic origin. They develop primarily inthe posterior section of the vocal cords in middle aged males and females. The granular cell tumours remain benign and can usually be extirpated endoscopically without recurrence. PMID- 6292141 TI - The role of consolidation irradiation in combined modality therapy of small cell carcinoma of the lung. AB - Forty-four patients with small cell carcinoma of the lung (SCCL) were treated with a program of combined chemotherapy and radiation therapy. Prophylactic cranial irradiation was given concurrent with the first of six planned cycles of chemotherapy consisting of Cyclophosphamide, Adriamycin, Vincristine and high dose Methotrexate (CAV-M). All patients judged as complete responders (CR) received consolidative thoracic irradiation (CTI) to the locoregional primary lung involvement. The CR rate to chemotherapy alone was 84% for patients with limited disease (LD) and 44% for extensive disease. In comparison to a prior trial, which used similar chemotherapy, but with irradiation withheld until primary site relapse, the actuarial primary site relapse rate at 2 years was reduced by CTI from 92% to 18% (P less than .01). The median primary site remission duration has not yet been reached in the CTI group and was 34 weeks without CTI (P less than .01). CTI increased the 2 year actuarial survival from 6% to 66% (P less than .01) in the chemotherapy CR patients. Median survival has not yet been reached in the CTI group, but was 48 weeks without CTI (P less than .01). Leptomeningeal spinal cord relapse in patients with no prior central nervous system (CNS) involvement occurred in 16% of patients relapsing. PMID- 6292142 TI - Special ovarian tumors and their management. PMID- 6292143 TI - Ovarian cancers in childhood. AB - In dealing with ovarian tumors in children, the gynecologist must use the same principle of assessing operability as those used for adults. This approach offers the maximum opportunity for successful treatment. The ancient medical philosophy of not inflicting harm if one cannot achieve good is especially applicable to children with ovarian cancer. It is better for a child to die at home in the loving care of her parents than alone in a cold, forbidding hospital room. There are certain characteristics that distinguish ovarian tumors in childhood from that in adults: (1) Limited space for tumor expansion. (2) A high degree of virulence in cancer. (3) A less effective immunologic response. (4) The possibility that therapy may affect future physical and emotional development. PMID- 6292144 TI - Integration of chemotherapy and radiation therapy for small cell carcinoma of the lung. AB - Two chemotherapy trials using cyclophosphamide, doxorubicin hydrochloride and high-dose vincristine sulfate with or without methotrexate have induced a 93% incidence of complete remission in limited disease presentation of small cell bronchogenic carcinoma of the lung and 39% incidence in extensive disease. The first trial without consolidation radiotherapy had a local failure rate of 65%, which dropped to 17% with consolidation radiotherapy to the primary and mediastinum. Prophylactic whole brain radiotherapy prevented local recurrence in 98% of evaluable patients. One carcinomatous meningitis and 5 intraspinal recurrences were noted among the 38 patients in the CAV-M trial. We conclude that high-dose vincristine sulfate is associated with an improved incidence of complete remission; that prophylactic whole brain radiotherapy has been highly successful; that prevention of intraspinal recurrence will necessitate the use of craniospinal axis radiation therapy and consolidation radiation therapy improves local control of primary and mediastinum. PMID- 6292145 TI - Compensation techniques in NIRS proton beam radiotherapy. AB - Proton beam has the dose distribution advantage in radiation therapy, although it has little advantage in biological effects. One of the best advantages is its sharp fall off of dose after the peak. With proton beam, therefore, the dose can be given just to cover a target volume and potentially no dose is delivered thereafter in the beam direction. To utilize this advantage, bolus techniques in conjunction with CT scanning are employed in NIRS proton beam radiation therapy planning. A patient receives CT scanning first so that the target volume can be clearly marked and the radiation direction and fixation method can be determined. At the same time bolus dimensions are calculated. The bolus frames are made with dental paraffin sheets according to the dimensions. The paraffin frame is replaced with dental resin. Alginate (a dental impression material with favorable physical density and skin surface contact) is now employed for the bolus material. With fixation device and bolus on, which are constructed individually, the patient receives CT scanning again prior to a proton beam treatment in order to prove the devices are suitable. Alginate has to be poured into the frame right before each treatments. Further investigations are required to find better bolus materials and easier construction methods. PMID- 6292146 TI - A bluetongue epizootic in northwestern United States. PMID- 6292148 TI - The mode of action of nanaomycins D and A on a gram-negative marine bacterium Vibrio alginolyticus. AB - Nanaomycin (NNM) D had a higher growth inhibitory activity than NNM-A against a Gram-negative marine bacterium, Vibrio alginolyticus. These quinone antibiotics were reduced by the respiratory chain-linked flavin dehydrogenase of the organism and the reduced forms of NNMs were quickly autoxidized by molecular oxygen to produce superoxide radicals (O2-). NNM-D was more effective than NNM-A both in the induction of KCN-insensitive oxygen consumption with the intact cells and in the production of O2- by the redox cycling. The growth inhibitory activities of NNM-D and A were partly reduced by raising the superoxide dismutase level of the cells. Thus, the ability to produce O2- at the cell membrane was correlated to the antibacterial activities of NNM-D and A. PMID- 6292147 TI - Disseminated histoplasmosis in a cat: successful treatment with ketoconazole. PMID- 6292149 TI - Enhancement of cerulenin production by a natural zeolite, an ammonium ion trapping agent. AB - Addition of a natural zeolite, a known ammonium ion-trapping agent, to a complex medium resulted in a several fold increase in cerulenin production by Cephalosporium caerulens. In the presence of zeolite, ammonium ion in the medium decreased, while mycelial growth increased to a small extent, and pH values remained constant. Small amounts of ammonium bicarbonate inhibited cerulenin production without affecting mycelial growth and pH values. It is thus assumed that zeolite stimulated cerulenin production by releasing the biosynthesis from the suppression by ammonium ions in C. caerulens. PMID- 6292150 TI - Induction of streptomycin-inactivating enzyme by A-factor in Streptomyces griseus. AB - The effect of A-factor on streptomycin resistance and productivity in Streptomyces griseus and S. bikiniensis was studied using A-factor-negative mutants. Resistance of several of these mutants was markedly increased by adding A-factor to the growing medium, as also was their streptomycin productivity. The A-factor induced resistance was due to inactivation by streptomycin-6 phosphotransferase, and enzyme synthesis in these mutants was completely dependent on the presence of A-factor. In the case of S. griseus 2247 where streptomycin productivity was independent of A-factor, resistance and synthesis of the inactivating enzyme were also independent of A-factor. A-Factor-negative mutants of S. griseus showed a decreased level of NADP-glycohydrolase and an increased level of several NADP-linked dehydrogenases, but these enzymes did not return to parental levels in cultures supplemented with A-factor. A-Factor seems to regulate streptomycin biosynthesis, not through an indirect metabolic sequence involving these enzymes but, more likely, by directly stimulating synthesis of enzyme(s) in the biosynthetic pathway. PMID- 6292152 TI - Biological activity of (5R,6S,8R)-6-alpha-hydroxyethyl-2-acetoxymethyl-2-penem-3 carboxylate. PMID- 6292151 TI - Studies on bacterial cell wall inhibitors. X. Properties of phosph-N acetylmuramoyl-pentapeptide-transferase in peptidoglycan synthesis of Bacillus megaterium and its inhibition by amphomycin. AB - The phospho-N-acetylmuramoyl-pentapeptide-transferase from Bacillus megaterium KM was characterized by the transfer reaction. The particulate enzyme preparation had the activity to transfer phospho-N-acetylmuramoyl-pentapeptide from UDP-N acetylmuramoyl-pentapeptide to undecaprenoid-1-ol-phosphate. The optimum pH for activity was about 8.5. The reaction required the presence of Mg2+ and an SH protector. With 25 mm Mg2+ the maximum activity was observed. The reaction was reversible and so the addition of UMP decreased the formation of undecaprenoid-1 ol-diphospho-N-acetylmuramoyl-pentapeptide. Amphomycin inhibited non competitively the transferase for the substrate UDP-N-acetylmuramoyl pentapeptide. PMID- 6292153 TI - Estimation of stoichiometric parameters for rumen fermentation of roughage and concentrate diets. PMID- 6292154 TI - Evaluation of various lignin assays for determining ruminal digestion of roughages by lambs. PMID- 6292155 TI - Purification and action sites of a follicle stimulating hormone inhibitor from bovine follicular fluid. AB - The purification and action sites of a follicle stimulating hormone (FSH) inhibitor obtained from bovine follicular fluid were examined. Bovine follicular fluid was salted out with ammonium sulfate as a concentration of 14.5 to 18.5%, and the salted out fraction was separated into two peaks by Sephadex G-200 column chromatography. The second peak, detectable as a single band by polyacrylamide gel disc electrophoresis, contained all the inhibitory activity to compensatory ovarian hypertrophy in mice. Serum FSH levels of unilaterally ovariectomized mice given injections of the inhibitor were lower than those of unilaterally ovariectomized mice given injections of saline. The inhibitor also suppressed FSH binding to granulosa cells in vitro. These results suggest that this inhibitor has two modes of action: suppression of FSH levels in serum and suppression of FSH binding to granulosa cells. PMID- 6292156 TI - Antifungals today. PMID- 6292157 TI - Penetration of cefotaxime and desacetylcefotaxime into skin blister fluid. PMID- 6292158 TI - Ceftriaxone: renal and biliary excretion and effect on the colon microflora. PMID- 6292160 TI - Sequence-specific DNA uptake in transformation of Neisseria gonorrhoeae. AB - Piliated, competent gonococci are known to preferentially take up homologous transforming DNA into the cell. We examined the mechanism for DNA uptake with pFA10, a hybrid 11.5-kilobase (kb) penicillin-resistant (Pcr) plasmid composed of heterologous DNA from a 7.2-kb Pcr plasmid and homologous DNA from a 4.2-kb gonococcal cryptic plasmid. The presence of the gonococcal cryptic plasmid DNA in the hybrid resulted in markedly increased transformation efficiencies in isogenic crosses as compared with the parent 7.2-kb Pcr plasmid. Uptake of 32P-end-labeled MspI or TaqI restriction fragments of the hybrid was limited to fragments entirely derived from the 4.2-kb gonococcal cryptic plasmid, indicating that DNA uptake was probably dependent on the presence of a specific DNA sequence. Since Haemophilus DNA did not inhibit transformation by the hybrid Pcr plasmid, the gonococcal DNA uptake sequence is different from the known sequence involved in homologous DNA uptake by Haemophilus spp. PMID- 6292159 TI - Insertions of transposon Tn5 into ribosomal protein PNA polymerase operons. AB - The genetic organization and interrelationships between the two ribosomal protein transcription units (the L11 and L10 operons) from near 89 min on the Escherichia coli chromosome were studied by using insertional mutations generated by the kanamycin-resistant transposable element Tn5. The polar effects of Tn5 insertions on the expression of the L11, L1, L10, and L12 ribosomal protein genes and the beta RNA polymerase subunit gene were examined (i) by the level of beta galactosidase activity generated from L10-lacZ and beta-lacZ gene fusions, (ii) by direct sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the proteins specified by plasmid ribosomal protein genes in UV-irradiated maxicells, and (iii) by urea-polyacrylamide gel electrophoresis of plasmid- and chromosome specified L12 protein. The results confirmed the organization of these genes into two transcription units as follows: PL11, rplK (L11), rplA (L1), PL10, rplJ (L10), rplL (L12), rpoB (beta). . .; they also localized the position of the PL10 promoter within an 80-nucleotide region near the end of the L1 gene. The results also support the idea that the translational regulatory proteins for the L11 and L10 operons are L1 and L10, respectively, and that the expression of the L12 gene is closely linked to L10 gene expression. PMID- 6292161 TI - Bioenergetic properties and viability of alkalophilic Bacillus firmus RAB as a function of pH and Na+ contents of the incubation medium. AB - The bioenergetic properties and viability of obligately alkalophilic Bacillus firmus RAB have been examined upon incubation in alkaline and neutral buffers in the presence or absence of added Na+. At pH 10.5, cells incubated in the absence of Na+ exhibited an immediate rise in cytoplasmic pH from less than 9.5 to 10.5, and they lost viability very rapidly. Viability experiments in the presence or absence of an energy source further suggested that the Na+-dependent mechanism for pH homeostasis is an energy-requiring function. The Na+/H+ antiporter, which catalyzes the vital proton accumulation at alkaline pH, was only slightly operational at pH 7.0; both whole cells and vesicles exhibited net proton extrusion even in the presence of Na+. Moreover, cells incubated in buffer at pH 7.0 were actually more viable in the presence of Na+ than in its absence. Thus, the inability of B. firmus RAB to grow at neutral pH is not due to excessive acidification of the cytoplasm. Rather, the transmembrane electrical potential, delta psi, generated at pH 7.0 was found to be much lower than at alkaline pH. The very low delta psi compromised several cell functions, e.g., Na+/solute symport and motility, which in this and other alkalophiles specifically depend upon delta psi and Na+. PMID- 6292162 TI - Enzymatic activities for interconversion of purines in spirochetes. AB - Enzymatic activities that catalyze the interconversion of purines and purine derivatives were detected in cell extracts of Spirochaeta aurantia, Spirochaeta stenostrepta, Treponema succinifaciens, and Treponema denticola. Phosphoribosyltransferase activities present in cell extracts of each of the four spirochete species functioned in the conversion of adenine, hypoxanthine, and guanine to AMP, IMP, and GMP, respectively. Nucleotidase activities in the extracts mediated the formation of nucleosides from nucleotides. The conversion of adenosine, inosine, and guanosine to the respective purine bases was catalyzed by nucleoside phosphorylase and, in some instances, by nucleoside hydrolase activities. Guanine deaminase activity was found in both S. aurantia and S. stenostrepta, whereas adenosine deaminase activity was detected only in S. aurantia. Adenine deaminase activity in T. succinifaciens extracts was sensitive to O2 and was relatively resistant to heating. Our results indicate that the four species of spirochetes studied possess a broad spectrum of purine interconversion enzymes. It is suggested that these enzymes may function in metabolic processes important for the survival of spirochetes in nutrient-poor natural environments. PMID- 6292163 TI - Cloning and expression of the beta-D-phosphogalactoside galactohydrolase gene of Lactobacillus casei in Escherichia coli K-12. AB - Lactose metabolism in Lactobacillus casei 64H is associated with the presence of plasmid pLZ64. This plasmid determines both phosphoenolpyruvate-dependent phosphotransferase uptake of lactose and beta-D-phosphogalactoside galactohydrolase. A shotgun clone bank of chimeric plasmids containing restriction enzyme digest fragments of pLZ64 DNA was constructed in Escherichia coli K-12. One clone contained the gene coding for beta-D-phosphogalactoside galactohydrolase on a 7.9-kilobase PstI fragment cloned into the vector pBR322 in E. coli strain chi 1849. The beta-D-phosphogalactoside galactohydrolase enzyme isolated from E. coli showed no difference from that isolated from L. casei, and specific activity of beta-D-phosphogalactoside galactohydrolase was stimulated 1.8-fold in E. coli by growth in media containing beta-galactosides. A restriction map of the recombinant plasmid was compiled, and with that information, a series of subclones was constructed. From an analysis of the proteins produced by minicells prepared from transformant E. coli cells containing each of the recombinant subclone plasmids, it was found that the gene for the 56-kilodalton beta-D-phosphogalactoside galactohydrolase was transcribed from an L. casei-derived promoter. The gene for a second protein product (43 kilodaltons) was transcribed in the opposite direction, presumably under the control of a promoter in pBR322. The relationship of this second product to the lactose metabolism genes of L. casei is at present unknown. PMID- 6292165 TI - Agrobacterium tumefaciens mutants affected in attachment to plant cells. AB - An analysis of Agrobacterium tumefaciens mutants with Tn5 insertions in chromosomal DNA showed that the chromosome of A. tumefaciens codes for a specific ability of this bacterium to attach to plant cells. This ability is associated with tumorigenesis by A. tumefaciens, the ability of avirulent A. tumefaciens to inhibit tumorigenesis, and the ability to adsorb certain phages. A second class of chromosomal mutations affects tumorigenesis without altering the ability to attach to plant cells. The attachment of A. tumefaciens to plant cells was assayed by mixing radiolabeled bacteria with suspensions of tobacco tissue culture cells or freshly isolated Zinnia leaf mesophyll cells. Under the conditions of this assay, an avirulent Ti plasmid-cured strain attached to the same extent as the same strain containing pTiB6806. Six of eight avirulent mutants with Tn5 insertions in chromosomal DNA showed defective attachment, whereas two retained wild-type attachment ability. In contrast to the strains showing wild-type attachment, the attachment-defective mutants failed to inhibit tumorigenesis when inoculated onto Jerusalem artichoke slices before inoculation of a virulent strain and also showed a loss of sensitivity to two Agrobacterium phages. The loss of phage sensitivity appeared to be due to a loss of ability to adsorb the phages. Staining with Calcofluor indicated that the mutants retained the ability to synthesize cellulose fibrils, which have been implicated in the attachment process. Southern filter hybridizations demonstrated that each mutant contained a single Tn5 insertion, and genetic linkage between the Tn5 insertion in one mutant and the attachment phenotype has also been demonstrated. PMID- 6292164 TI - Mapping of Streptococcus faecalis plasmids pAD1 and pAD2 and studies relating to transposition of Tn917. AB - Plasmids pAD1 (37.8 megadaltons) and pAD2 (17.1 megadaltons) of Streptococcus faecalis strain DS16 have been mapped with restriction enzymes. The location of a hemolysin-bacteriocin determinant on the conjugative pAD1 plasmid was derived from analyses of transposon insertions. Electron microscope and hybridization analyses located Tn917(Em) and the streptomycin (Sm) and kanamycin (Km) resistance determinants on the nonconjugative pAD2 plasmid. It was shown previously that the erythromycin (Em) resistance associated with Tn917 is inducible and that transposition from pAD2 to pAD1 is also stimulated by exposure of cells to low concentrations of Em. Here we show that inducing concentrations of Em also increase the conjugative transfer potential of pAD1; this is possibly related to a mild and short-lived inhibitory stress placed on the cells before full induction of resistance. Selection of Em-resistant transconjugants arising from matings between DS16 and a plasmid-free recipient gave rise to transconjugants which primarily harbor stable pAD1::pAD2 cointegrates. A 30-min exposure of donors to Em (0.5 microgram/ml) before mating resulted in a severalfold increase in the number of such transconjugants. However, a small fraction (e.g., 3 of 40) of these Emr Smr Kmr transconjugants harbored pAD1::Tn917 and pAD2 molecules. Since we believe pAD2 is incapable of being mobilized by pAD1 without being covalently linked, it is likely that transfer in these cases involved cointegrates representing structural intermediates in the transposition of Tn917 from pAD2 to pAD1. It follows that such intermediates probably had two copies of Tn917 and readily resolved after transfer. (These cointegrates are different from the stable cointegrates which were shown to have only a single copy of Tn917; the latter are assumed not to be related to transposition.) Two variants with altered Tn917 transposition properties were derived. One of them transposed at an elevated frequency, whereas the other showed no detectabel transposition. In neither case was transposition influenced by Em exposure; however, both remained inducible for Em resistance. PMID- 6292166 TI - Physical mapping of TOL plasmids pWWO and pND2 and various R plasmid-TOL derivatives from Pseudomonas spp. AB - Analysis of several independently isolated R plasmid-TOL hybrids revealed a wide variation in the amount of TOL DNA they contain. If the formation of the various R plasmid-TOL hybrids involves transposition (which has yet to be rigorously assessed), such transposition does not involve a unique segment of TOL DNA. PMID- 6292167 TI - Restriction enzyme mapping of the DNA of Streptomyces bacteriophage B alpha and its deletion derivatives. AB - Cleavage analysis of actinophage B alpha DNA was done with several restriction enzymes, and a restriction map of the DNA was determined. The DNA appeared to carry cohesive ends. Deletion mutants of actinophage B alpha were isolated by five cycles of treatment with 15 mM PPi. Both mutants had deletions of 2.5 of 1.8 megadaltons near one end of the genome, and one of them lost the single EcoRI cleavage site. PMID- 6292168 TI - Adenosine kinase-deficient mutant of Neurospora crassa. AB - Tubercidin-resistant mutant strains of Neurospora crassa were isolated, and at least one appeared to be deficient in adenosine kinase. No significant differences in [8-14C]adenosine labeling of purine nucleotides or nucleosides were found between the wild type and the adenosine kinase-deficient strains. PMID- 6292169 TI - Glycerol kinase as a substitute for dihydroxyacetone kinase in a mutant of Klebsiella pneumoniae. AB - With dihydroxyacetone as the sole source of carbon and energy, constitutively synthesized glycerol kinase of the glp system supported aerobic growth of Klebsiella pneumoniae mutants lacking the inducible dihydroxyacetone kinase of the dha system. Glycerol kinase had an apparent Km of 0.01 mM for its physiological substrate and 1 mM for its surrogate substrate. However, the growth rate on dihydroxyacetone of cells relying on glycerol kinase increased with the concentration of the carbon and energy source up to 50 mM, suggesting that permeation is rate limiting. PMID- 6292171 TI - Salt-concentration dependence of thermal denaturation of restriction fragment DNAs from phi X174. AB - High-resolution differential melting curves of phi X174 Y1 and Y2 restriction fragment DNAs, for which the base sequences were known, were measured at various sodium ion concentrations ranging from 195 to 2.3 mM. The curves were resolved into component peaks, and the change in the melting temperature, the change in the area, and the change in the breadth of each peak with change in salt concentration were examined. The locations of the melting regions corresponding to the peaks in the melting curves were assigned based on theoretical calculations of melting curves and stability maps. It was found that as the salt concentration was decreased from the high to the intermediate range, the breadths of the peaks on the low-temperature side decreased whereas those on the high temperature side remained almost constant, and also the separation between the peaks along the temperature axis increased. Changes in the positions of peaks relative to one another were interpreted in terms of the difference in the free energy increase between a loop state and an end-coil state as the salt concentration decreased. PMID- 6292170 TI - Chromosomal replication origins (oriC) of Enterobacter aerogenes and Klebsiella pneumoniae are functional in Escherichia coli. AB - The chromosomal DNA replication origins (oriC) from two members of the family Enterobacteriaceae, Enterobacter aerogenes and Klebsiella pneumoniae, have been isolated as functional replication origins in Escherichia coli. The origins in the SalI restriction fragments of 17.5 and 10.2 kilobase pairs, cloned from E. aerogenes and K. pneumoniae, respectively, were found to be between the asnA and uncB genes, as are the origins of the E. coli and Salmonella typhimurium chromosomes. Plasmids containing oriC from E aerogenes, K. pneumoniae, and S. typhimurium replicate in the E. coli cell-free enzyme system (Fuller, et al., Proc. Natl. Acad. Sci. U.S.A. 78:7370--7374, 1981), and this replication is dependent on dnaA protein activity. These SalI fragments from E. aerogenes and K. pneumoniae carry a region which is lethal to E. coli when many copies are present. We show that this region is also carried on the E. coli 9.0-kilobase pair EcoRI restriction fragment containing oriC. The F0 genes of the atp or unc operon, when linked to the unc operon promoter, are apparently responsible for the lethality. PMID- 6292173 TI - Effects of temperature and cholesterol on human erythrocyte membranes. AB - The effects of temperature and cholesterol on the membrane fluidity of human erythrocytes were studied using 5-nitroxide stearic acid (5NS), 12-nitroxide stearic acid (12NS), and 16-nitroxide stearic acid (16NS). Human erythrocytes and their lipid vesicles were treated in the range of 5--55 degrees C. In erythrocytes, ESR signals for 12NS and 16NS showed line broadening above 40 degrees C, whereas those for 5NS became sharper with increasing temperature as was the case with the signals of lipid vesicles for each label molecule. Lipid extraction from the heated sample caused no radical reduction. Only in 12NS labeled erythrocytes did a weakly immobilized component and a strongly immobilized component appear. In the time course at 50 degrees C, the former decreased and the latter remained constant. From the ratio of both components, it was found that the interaction of the label molecules with the binding sites was determined by the physical state of the membrane. Furthermore, the dependence on temperature of the molecular motion of the labels in the cell membrane was irreversible above 40 degrees C. On addition of cholesterol to the membrane, the outer hyperfine splittings for 12NS and 16NS increased but that for 5NS decreased at C/P greater than 1, perhaps indicating a spread between the head groups of phospholipids by cholesterol. PMID- 6292172 TI - Acid sphingomyelinase of human placenta: purification, properties, and 125iodine labeling. AB - Human placental acid sphingomyelinase was highly purified in the presence of Triton X-100. DEAE-Sephacel chromatography and chromatofocusing were the most effective steps in the purification procedure. Enzyme purification was 380,000 nmol/mg protein/h. Characterization and radioiodination were carried out with the chromatofocusing fraction containing highly purified enzyme. The purified enzyme contained no activity of eleven other lysosomal hydrolases but hydrolyzed bis-p nitrophenyl phosphate slowly compared with [14C]sphingomyelin and chromogenic substrates. SDS-gel electrophoresis revealed two distinct protein bands with molecular weights of 70,500 and 39,800. This enzyme had a molecular weight of 200,000 as determined by analytical gel filtration. The pH optimum was 5.0 and Km was 52.6 x 10(-5) M for [14C]sphingomyelin. Highly purified sphingomyelinase was labeled with 125iodine by the use of Enzymobeads. Labeled sphingomyelinase preparation was rapidly cleared from blood with t1/2 of 1 min. It was absorbed mostly into the liver and presumably largely excreted from there. This labeled enzyme may be useful in metabolic studies in normal animals and animal models of genetic lysosomal storage disorders. PMID- 6292174 TI - Purification and properties of magnesium- and manganese-dependent ribonucleases H from chick embryo. AB - Two RNases H, Mg2+- and Mn2+-dependent RNases H, are present in extracts of chick embryo. These RNases H can be separated by phosphocellulose column chromatography. Mg2+-dependent RNase H was purified over 900-fold and Mn2+ dependent RNase H over 1,700-fold from chick embryo extracts. The molecular weight of the purified Mg2+-dependent RNase H was about 40,000 and of the Mn2+ dependent RNase H about 120,000, when estimated by gel filtration. Mg2+-dependent RNase H exhibits maximal activity at pH 9.5, and requires 15 to 20 mM Mg2+ for maximal activity, whereas Mn2+-dependent RNase H is most active at pH 8.5, and is maximally active at the concentration of 0.4 mM Mn2+, and has some activity with Mg2+. Both enzymes require a sulfhydryl reagent for maximal activity. Mn2+ dependent RNase H was inhibited by o-phenanthroline, pyrophosphate, and those polyamines tested, whereas Mg2+-dependent enzyme was not, although it was inhibited by NaF. Both RNases H liberate a mixture of oligonucleotides with 5' phosphate and 3'-hydroxyl termini endonucleolytically. PMID- 6292175 TI - Differential exposure of components of cytochrome b-c1 region in beef heart mitochondria and electron transport particles. AB - The reduction of cyctochromes c + c1 by durohydroquinone and ferrocyanide in electron transport particles (ETP) and intact cytochrome c-depleted beef heart mitochondria has been studied. At least 94% of the ETP are in an inverted orientation. Durohydroquinone reduces 80% of c + c1 in ETP but less than 20% in mitochondria; sonication of mitochondria allows reduction of cytochromes c + c1 (80%). Addition of ferrocyanide (effective redox potential +245 mV) to electron transport particles results in 30% reduction of cytochromes c + c1. Addition of ferrocyanide to intact cytochrome c-depleted mitochondria does not reduce cytochrome c1; treatment with N,N,N',N'-tetramethylphenylenediamine, Triton X 100, or sonic oscillation results in 30% reduction of cytochromes c + c1. The Km value of ferrocyanide oxidase for K-ferrocyanide is pH-dependent in ETP only, increasing with increasing pH. The extent of reduction of cytochrome c1 is also pH-dependent in ETP only, the extent of reduction increasing with decreasing pH. On the basis of these data cytochrome c1 is exposed to the matrix face and cytochrome c is exposed to the cytoplasmic face. No redox center other than cytochrome c in the segment between the antimycin site and cytochrome c is exposed on the C-side. PMID- 6292176 TI - The role of the adenine nucleotide translocator in oxidative phosphorylation. A theoretical investigation on the basis of a comprehensive rate law of the translocator. AB - A minimum model of adenine nucleotide exchange through the inner membrane of mitochondria is presented. The model is based on a sequential mechanism, which presumes ternary complexes formed by binding of metabolites from both sides of the membrane. The model explains the asymmetric kinetics of ADP-ATP exchange as a consequence of its electrogenic character. In energized mitochondria, a part of the membrane potential suppresses the binding of extramitochondrial ATP in competition with ADP. The remaining part of the potential difference inhibits the back exchange of internal ADP for external ATP. The assumption of particular energy-dependent conformational states of the translocator is not necessary. The model is not only compatible with the kinetic properties reported in the literature about the adenine nucleotide exchange, but it also correctly describes the response of mitochondrial respiration to the extramitochondrial ATP/ADP ratio under different conditions. The model computations reveal that the translocation step requires some loss of free energy as driving force. The size of the driving force depends depends on the flux rate as well as on the extra- and intramitochondrial ATP/ADP quotients. By both quotients the translocator control the export of ATP formed by oxidative phosphorylation in mitochondria. PMID- 6292177 TI - Expression of an Abelson murine leukemia virus-encoded protein in Escherichia coli causes extensive phosphorylation of tyrosine residues. AB - A segment of the Abelson murine leukemia virus (A-MuLV) genome was inserted into an Escherichia coli plasmid designed to allow the expression of the protein encoded by the viral gene. Bacteria expressing the A-MuLV-encoded protein were isolated; they had new phosphorylated proteins in which the phosphate was linked to tyrosine residues. These proteins included many that must be E. coli protein. One phosphotyrosine-containing protein of 62,000 molecular weight had reactivity with antiserum specific for authentic A-MuLV protein. The A-MuLV protein thus appears to be a tyrosine-specific protein kinase which is active in E. coli. PMID- 6292178 TI - ATP-dependent lysosomal cystine efflux is defective in cystinosis. AB - Lysosomes containing large amounts of the amino acid, cystine, were obtained from transformed, cultured, human lymphoblasts which had been exposed to cystine dimethyl ester. Lysosomal cystine efflux was greatly enhanced by exogenous ATP in cell lines from normal individuals. Cystine efflux was unresponsive to ATP in lysosomes from individuals with the disorder, cystinosis. Efflux of cystine from normal cell lysosomes was inhibited by both the ATP analog, 5 adenylylimidodiphosphate, and the proton translocator, carbonyl cyanide m chlorophenylhydrazone. Efflux was not affected by ouabain or oligomycin. Thus, lysosomal cystine efflux is dependent upon the functioning of a proton-pump ATPase. ATPase-dependent cystine efflux appears to be aberrant in cystinotic cell lysosomes. PMID- 6292179 TI - Calcium antagonists High-affinity binding and inhibition of calcium transport in a clonal cell line. AB - PC12 cells, a clonal rat pheochromocytoma cell line, possess voltage-dependent calcium channels that bind the high affinity dihydropyridine calcium antagonist [3H]nitrendipine and other calcium channel blockers. The binding is temperature dependent and saturable, and shows no cooperativity. The calcium antagonists inhibit potassium-induced 45Ca uptake into the cells with approximately the same potencies as those needed to inhibit [3H]nitrendipine binding to cell membranes. The affinity of these compounds for the PC12 cell calcium channel is slightly lower than that reported for binding to brain and heart. Potassium-stimulated 45Ca uptake into PC12 cells is rapid, being half-maximal within 30 s at 20 degrees C. Different classes of calcium antagonists seem to block calcium flux at different sites on the calcium channel. A lower limit of the rate of calcium movement through a single channel is given. PC12 cells seem to be a suitable model system for the study of the pharmacology and biochemistry of the voltage dependent calcium channel. PMID- 6292180 TI - Down-regulation of the interferon receptor. AB - The binding of 125I-labeled alpha A interferon to human lymphoblastoid Daudi cells decreased when these cells were incubated with unlabeled alpha or beta interferon. This decrease could not be accounted for by the occupancy of interferon receptors with unlabeled interferon and it apparently resulted from the loss or down-regulation of receptors. The binding activity gradually increased when Daudi cells were incubated in fresh medium after a treatment with interferon, but inhibition of protein synthesis with cycloheximide prevented this recovery. Treatment of Daudi cells with this inhibitor resulted in the loss of half the interferon binding activity within 5 h. These findings suggested that the interferon receptors turn over at a basal rate in interferon-free medium and at an increased rate in cells incubated with interferon. The dose-response for the down-regulation was investigated by treating Daudi cells with different concentrations of alpha interferon. Down-regulation was observed in cells treated with relatively low doses of interferon, sufficient to elicit a biological response. The synthesis of the enzyme (2',5')oligo(A) polymerase was induced at the lowest interferon concentrations tested which caused receptor down regulation. PMID- 6292181 TI - A kinetic analysis of the interaction of human myeloperoxidase with hydrogen peroxide, chloride ions, and protons. AB - The effect of H2O2, Cl-, and pH on human myeloperoxidase activity has been examined. The Km for H2O2 is shown to be affected by the combined presence of Cl- and acid pH conditions. The Km for H2O2 is independent of pH in the absence of Cl and dependent on pH in the presence of Cl-. Conversely, the dependence of the Km for H2O2 on Cl- concentration increases as the pH decreases. A model is proposed in which Cl- has a dual role, acting both as a substrate and as an inhibitor. According to this model, the inhibitor Cl- binding site must be protonated prior to the binding of Cl- and is distinct from the substrate Cl- binding site which is unaffected by pH. The rate equation derived from this model is used to further analyze the data presented. The values of Km for H2O2 predicted by the rate equation are in good agreement with the experimentally determined values. PMID- 6292182 TI - Probing of the coupling site of the beta-adrenergic receptor. Competition between different forms of the guanyl nucleotide binding protein for interaction with the receptor. AB - The interaction of the beta-adrenergic receptor (R) with different forms of the regulatory protein (G) was studied. For this purpose, the reconstituted system formed from separate soluble preparations of R and G was employed (Citri, Y., and Schramm, M. (1980) Nature (Lond.) 287, 297-300). Soluble preparations of the native nonactive G, to which GDP is bound (GGDP), and the persistently active G, to which guanyl-5'-yl-imidophosphate (Gpp(NH)p) is bound GGPP(MH)P), were prepared. Reconstituted systems made of GGDP, GGPP(NH)P, and R were used to test whether each of the two forms of G can affect the other's interaction with R. GGPP(NH)P was found to compete with GGDP for the interaction with R. However, GGDP was unable to interfere with the interaction of R with GGPP(NH)P. Similar experiments were carried out with another persistently active G (GGTP gamma S). In sharp contrast to GGPP(NH)P, GGTP gamma S was found incapable of competing with GGDP for R. The above experiments suggest that GGDP and GGPP(NH)P interact with the same coupling site on R. However, the degree of recognition of G by R depends strongly on the guanyl nucleotide which is bound to G. The relative order of recognition being GGPP(NH)P greater than GGDP greater than GGTP gamma S approximately 0. These findings may also explain the known irreversible effects of GTP gamma S in the adenylate cyclase system. PMID- 6292183 TI - Identification of cGMP-stimulated cyclic nucleotide phosphodiesterase in lung tissue with monoclonal antibodies. AB - Monoclonal antibodies (CGS-1, 2, 3, and 4) and rabbit antisera have been produced against bovine heart cyclic GMP-stimulated cyclic nucleotide phosphodiesterase. The monoclonal antibodies were all of the IgG1 subclass and had relatively high affinities (kd values, 1-5 X 10(-10) M), making them suitable for many sensitive immunological and analytical procedures. Most of the monoclonal antibodies did not inhibit the enzyme, which allowed a rapid and specific assay for this form of phosphodiesterase to be developed. All of the activity could be adsorbed from purified preparations of cGMP-stimulated phosphodiesterase by each of the antibodies indicating antigenic homogeneity of the cGMP-stimulated phosphodiesterase. Neither the monoclonal antibodies nor the antisera inhibited cGMP-binding to the phosphodiesterase. Substrate specificity, Electrophoresis, and cyclic GMP binding studies carried out on immunoadsorbed material indicated that a major part of the cyclic nucleotide phosphodiesterase activity present in bovine lung was due to the cyclic GMP-stimulated form. The presence of this enzyme was difficult to identify by other more common procedures and has not been previously demonstrated in this tissue. These studies also indicated that the cGMP-stimulated phosphodiesterase was immunologically distinct from the other major cGMP-binding proteins present in lung extract and that it could be separated from these binding proteins by chromatography on DEAE-cellulose. Finally, the data suggest that none of the other peaks of cyclic nucleotide phosphodiesterase activity, seen upon DEAE-cellulose fractionation of lung extracts, contain the immunological determinants present on the cGMP-stimulated form which are recognized by the antibodies. PMID- 6292184 TI - Interaction of interferon with cellular receptors. Internalization and degradation of cell-bound interferon. AB - Human interferon alpha A, produced in Escherichia coli by recombinant DNA technology, was labeled with 125I to study its binding to receptors on human lymphoblastoid Daudi cells. This binding showed a marked temperature dependency, with maximum binding obtained at 30-37 degrees C. About 60% of the cell-bound radioactivity was released upon subsequent addition of unlabeled interferon, indicating that only part of the cell-bound interferon could be displaced by competitor. Moreover, about 30-50% of cell-bound interferon was not released by treating the cells with 0.2 N acetic acid, a procedure which removes polypeptide hormones on the cell surface, indicating that part of the interferon bound at 37 degrees C was internalized. This interferon was slowly degraded to acid-soluble products, which were released into the culture medium. Treatment of DAudi cells with the lysosomotropic amines chloroquine and methylamine inhibited the degradation of interferon. Methylamine, however, also inhibited the internalization of interferon. Daudi cells treated with interferon in the presence of chloroquine showed an increase in the interferon-induced enzyme 2',5' oligo(A) polymerase comparable to that of cells treated with interferon alone. This enzyme increased to a similar extent in cells treated with interferon and cytochalasin, a drug which inhibited internalization of interferon by 50%. These results suggest that degradation and possibly internalization of interferon are not required for at least some of its biological activities. PMID- 6292185 TI - Internalization and degradation of receptor-bound human choriogonadotropin in Leydig tumor cells. Fate of the hormone subunits. AB - The studies presented herein were aimed at characterizing the pathway involved in the internalization and degradation of human choriogonadotropin by cultured Leydig tumor cells. A quick biochemical method that differentiates between the surface-bound and internalized hormone was developed. Using this method and two hormone derivatives labeled exclusively (with 125I) in the alpha or beta subunits, it was possible to follow the fate of each hormone subunit during hormone binding, internalization, and degradation. The results show that the hormone is internalized in the intact form and that it reaches its place of degradation (presumably the lysosomes) in the intact form. The pathway for degradation of the internalized hormone is complex, and it appears to involve processing of one or both subunits of the intact hormone, followed by subunit dissociation and further degradation of the individual subunits. The alpha subunit is quickly degraded by the cells. The only detectable degradation products are extracellular amino acids. The beta subunit is degraded slower, and several intracellular degradation products are detectable before amino acids appear in the medium. PMID- 6292186 TI - Identification of the endogenous retinoids associated with three cellular retinoid-binding proteins from bovine retina and retinal pigment epithelium. AB - The endogenous retinoids associated with three cellular retinoid-binding proteins from bovine retina and retinal pigment epithelium have been identified by their spectral characteristics and their co-migration with authentic retinoids on high performance liquid chromatography. All-trans-retinol is the only retinoid associated with the cellular retinol-binding protein from retina and retinal pigment epithelium. The saturation of the binding site with native ligand is 0.95 +/- 0.05 mol of retinoid/mol of protein for cellular retinol-binding protein purified from frozen and fresh retina and retinal pigment epithelium. All-trans retinoic acid has been identified as the endogenous ligand associated with the cellular retinoic acid-binding protein of cattle retina. The saturation of the binding site of cellular retinoic acid-binding protein from frozen and fresh retina (the protein is absent in extracts of retinal pigment epithelium) is about 0.2 mol of retinoid/mol of protein, probably a minimum value due to losses of the ligand. Retinoic acid has not been detected previously in retina. Cellular retinaldehyde-binding protein from retina purifies with two endogenous ligands which cochromatograph on high performance liquid chromatography with 11-cis retinaldehyde and 11-cis-retinol and occur in a ratio of approximately 3:1, respectively. The binding site of cellular retinaldehyde-binding protein from frozen retina is nearly fully occupied with these two ligands (saturation greater than 0.9 mol of retinoids/mol of protein). In contrast, purified cellular retinaldehyde-binding protein from retinal pigment epithelium carries only 11-cis retinaldehyde as an endogenous ligand. The saturation of the binding site with this ligand is greater than 0.95. PMID- 6292187 TI - New fluorescent analogs of cAMP and cGMP available as substrates for cyclic nucleotide phosphodiesterase. AB - The synthesis of fluorescent derivatives of cAMP and cGMP, by reaction with isatoic anhydride in aqueous solution at mild pH and temperature, yielding 2'-O anthraniloyl derivatives of cyclic nucleotides, is here described. 2'-O-(N Methylanthraniloyl) derivatives were also synthesized by reaction with N methylisatoic anhydride. Upon excitation at 330-350 nm, these derivatives exhibited maximum fluorescence emission at 430-445 nm in aqueous solution with quantum yields of 0.11-0.26. Their fluorescence was sensitive to the polarity of solvent; in N,N-dimethylformamide quantum yields of 0.8-0.95. The major differences between the two fluorophores were the longer wavelength of the emission maximum of the N-methylanthraniloyl group and its greater quantum yield. The derivatives were substrates for beef heart cyclic nucleotide phosphodiesterase, 15-24% as effective as the natural substrate cAMP. When combined with thin layer chromatography techniques, two apparent Km values (3-4 microM and 36-76 microM) for the cAMP derivatives and one value (10-18 microM) for the cGMP derivatives were obtained. The results indicate that these 2' hydroxyl-modified cAMP and cGMP can be useful fluorescent substrate analogs for cyclic nucleotide phosphodiesterase. PMID- 6292188 TI - Phosphorylation and activation of rabbit skeletal muscle phosphorylase kinase by a cyclic nucleotide- and Ca2+-independent protein kinase. AB - Phosphorylase kinase from rabbit skeletal muscle can be phosphorylated and activated by a cyclic nucleotide- and Ca2+-independent protein kinase previously identified as a glycogen synthase kinase (Itarte, E., and Huang, K.-P. (1979) J. Biol. Chem. 254, 4052-4057). This independent kinase phosphorylates the beta subunit of phosphorylase kinase approximately 15 times faster than it does the alpha subunit. The cAMP-dependent and -independent kinases separately catalyze the incorporation of 1 mol of phosphate into the beta subunit. Analyses of the tryptic peptides from the beta subunit phosphorylated with either kinase by isoelectric focusing and peptide mapping indicate that both kinases phosphorylate the same site on the beta subunit. Activation of phosphorylase kinase catalyzed by the independent kinase is only 60% of that observed with cAMP-dependent kinase. If phosphorylase kinase is first incubated with the independent kinase to phosphorylate the beta subunit, subsequent addition of cAMP-dependent kinase results in a predominant phosphorylation of the alpha subunit. This additional phosphorylation of the alpha subunit is accompanied by a further activation of the alpha subunit is accompanied by a further activation of phosphorylase kinase to the same extent as that achieved by cAMP-dependent kinase alone. Hence, the phosphorylation of the alpha subunit is clearly required for full activation of phosphorylase kinase, even at low [Mg2+]. PMID- 6292189 TI - Radioimmunochemical determination of cellular retinol- and cellular retinoic acid binding proteins in cytosols of rat tissues. AB - Radioimmunoassays have been developed for cellular retinol-binding protein and cellular retinoic acid-binding protein, postulated mediators of vitamin A action in nonvisual functions. These assays are considerably more sensitive for detection and quantitation of these proteins than previous methods. The higher sensitivity has permitted the detection of cellular retinoic acid-binding protein in adult rat tissues previously considered negative. These include kidney, lung, and spleen. The only tissues negative for both binding proteins were adrenals, skeletal muscle, ileal mucosa, and serum. Assay of tissue levels of cellular retinol-binding protein and cellular retinoic acid-binding protein reveals them to be widely distributed throughout most organs of the rat. No tissue or sex differences in these proteins were detected in these studies. PMID- 6292190 TI - Regulation of phosphoenolpyruvate carboxykinase (GTP) synthesis in rat liver cells. Rapid induction of specific mRNA by glucagon or cyclic AMP and permissive effect of dexamethasone. AB - Isolated rat liver cells maintained in suspension culture for 4 to 5 h synthesize the gluconeogenic cytosolic enzyme phosphoenolpyruvate carboxykinase at a rate approximately 5-fold lower than the in vivo hepatic rate. Glucagon rapidly re induces phosphoenolpyruvate carboxykinase synthesis in such cells. The rate of enzyme synthesis doubles in 40 min and plateaus at a level 6- to 13-fold higher than in control cells 120 min after glucagon addition at maximal concentration. Consistent with the presumed role of cyclic AMP as a mediator of enzyme induction, the phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine, added simultaneously with glucagon, shifts the hormone dose-response curve 2 log units to the left. Moreover, cyclic AMP supplied exogenously to the cells mimics the inductive effect of glucagon. Total cellular RNA isolated from hepatocytes induced by glucagon contains an increased level of mRNA coding for phosphoenolpyruvate carboxykinase, as determined by translational assay. The kinetics and extent of the rise in mRNA level are adequate to explain the stimulation of enzyme synthesis. Although glucagon on its own induces a build-up of phosphoenolpyruvate carboxykinase mRNA and a commensurate stimulation of enzyme synthesis, the glucagon induction is very markedly amplified when the cells are first preincubated with dexamethasone. The glucocorticoid by itself, however, does not have any substantial effect on the level of phosphoenolpyruvate carboxykinase mRNA or on the rate of enzyme synthesis. Its role can therefore be characterized as permissive. PMID- 6292191 TI - Coupling of voltage-sensitive sodium channel activity to stretch-induced amino acid transport in skeletal muscle in vitro. PMID- 6292192 TI - Models of the two heme centers in cytochrome oxidase. The optical properties of cytochrome a and a3. AB - High and low spin complexes of ferric and ferrous heme a have been prepared and characterized spectroscopically. Bis(1-methylimidazole) heme a provides a good model for cytochrome a in both oxidation states while several spectral properties of cytochrome a3 can be reproduced by 1,2-dimethylimidazole heme a3. The visible absorbance spectra of these analogs account well for the absorbance spectra of oxidized and reduced cytochrome oxidase and support the conclusion (Vanneste, W. (1966) Biochemistry 5, 838-848) that cytochrome a provides the major contribution to the spectral changes in the 600 nm band upon reduction. The 655 nm band present in cytochrome oxidase appears to be a characteristic of high spin heme a+3. PMID- 6292193 TI - Stereospecific removal of the DR hydrogen atom at the 10-carbon of arachidonic acid in the biosynthesis of leukotriene A4 by human leukocytes. PMID- 6292194 TI - Regulation of human lymphoblast plasma membrane 5'-nucleotidase by zinc. PMID- 6292195 TI - Hormonal inhibition of adenylate cyclase. alpha 2 Adrenergic receptors promote release of [3H]guanylylimidodiphosphate from platelet membranes. PMID- 6292196 TI - Glucocorticoid regulation of 1,25(OH)2-vitamin D3 receptors in cultured mouse bone cells. PMID- 6292197 TI - Specific binding of prostaglandin D2 to rat brain synaptic membrane. Occurrence, properties, and distribution. AB - Prostaglandin (PG) D2 bound specifically to a particulate fraction rich in the synaptic membrane of rat brain. The binding was dependent on time and temperature, equilibrium being reached after 5 min at 37 degrees C. The specific binding constituted about 70% of the total binding at 37 degrees C, and 55% at 0 degrees C. The maximal binding was obtained in the presence of 100 mM sodium ion and at pH 8. The equilibrium dissociation constant and the maximal concentration of binding sites as determined by Scatchard analysis were 28 +/- 7 nM and 0.45 pmol/mg of protein (n = 3), respectively. Hill coefficient was 1.15, indicating a single entity of binding sites and no cooperativity. The binding sites were highly specific for PGD2; the Ki values for PGD1 and PGF2 alpha were 523 and 693 nM, respectively. Other PGs including 13,14-dihydro-15-keto-PGD2, an inactive metabolite of PGD2, had 150- to 1000-fold lower affinities than PGD2. The binding was inhibited by boiling or treatment with proteases, phospholipases, or beta galactosidase. The specific activity of PGD2 binding was highest in the pituitary gland, followed by the hypothalamus and the olfactory bulb od the rat brain, this pattern being almost parallel to that of the cytosolic NADP-linked PGD2 dehydrogenase activity. The results suggest that PGD2 plays a significant role in these regions of the rat brain. PMID- 6292198 TI - Phosphorylative modification of histone H1 subspecies following isoproterenol and N6,O2'-dibutyryl cyclic AMP stimulation of rat C6 glioma cells. PMID- 6292199 TI - Osmoregulation of gene expression. I. DNA sequence of the ompR gene of the ompB operon of Escherichia coli and characterization of its gene product. AB - The ompB region on the Escherichia coli chromosome codes for two genes, ompR and envZ, which are required for the osmolarity sensitive biosynthetic regulation of the outer membrane matrix proteins (porins), OmpF and ompC. A part of the ompB region containing the ompR gene has been cloned (Wurtzel, E. T., Movva, N. R., Ross, F. L., and Inouye, M. (1981) J. Mol. Appl. Genet. 1, 61-69). We have determined the DNA sequence, including the promoter and structural regions encompassed in a 1.3-kilobase pair Ava I-Eco RI subfragment. This fragment codes for the entire ompR gene as well as the 5' end of the envZ gene. The ompR gene codes for a protein of 32,489 daltons, consisting of 284 amino acid residues. This was confirmed by identifying the gene product by sodium dodecyl sulfate polyacrylamide gel electrophoresis and determining a partial amino acid sequence of the NH2-terminal region of the gene product. A sequence of 57 amino acid residues located in the COOH-terminal region of the protein is extremely basic. It contains 10 arginine plus lysine residues in contrast to 1 glutamic acid residue in this region. In vitro transcription of the DNA from this region indicates that ampR and envZ are co-transcribed as a polycistronic mRNA from a promoter located 5' to the ompR gene. Translation of the am pR gene terminates at two tandem TAS codons and translation of the envZ gene initiates 29 nucleotides downstream. Cloning of the promoter region of ompB at a site 5' to the structural portion of the beta-galactosidase gene indicates that transcription of ompB is under positive control by cAMP. PMID- 6292200 TI - Osmoregulation of gene expression. II. DNA sequence of the envZ gene of the ompB operon of Escherichia coli and characterization of its gene product. PMID- 6292201 TI - Oxidation of Escherichia coli iron centers by the myeloperoxidase-mediated microbicidal system. AB - Myeloperoxidase, HeO2, and a halide (chloride, bromide, iodide) constitute a powerful microbicidal system which is active against a wide variety of microorganisms and is believed to contribute to the antimicrobial activity of neutrophils. The precise mechanism by which this system exerts its toxicity is unknown. We report here that the microbicidal activity of the myeloperoxidase-H2O chloride system on Escherichia coli is associated with the loss of iron into the medium as measured by the release of 59Fe from prelabeled organisms. Iron loss (but not bactericidal activity) was considerably increased by the addition of EDTA or other iron chelators; it was not associated with a corresponding release of protein with 14C-amino-acids. Iron loss was observed with chloride or bromide as the halide, but not when iodide was employed in microbicidal concentrations. Microbicidal activity was detected at an earlier time period and at a lower halide concentration than was iron loss. Analogous changes were observed when cytochrome c was oxidized by the myeloperoxidase H2O2-halide system. The initial response was a shift in the Soret maximum, followed by a fall in absorbance accompanied by the loss of iron. As with the intact organism, iron loss was evident with chloride and bromide, but not with iodide as the halide. These findings suggest that microbial iron centers are a target for the myeloperoxidase mediated antimicrobial system and that their oxidation may contribute to microbicidal activity. PMID- 6292202 TI - Stimulation of specific 1,25-dihydroxyvitamin D3 binding protein in cultured postnatal rat intestine by hydrocortisone. PMID- 6292203 TI - The Escherichia coli dnaC gene product. I. Overlapping of the dnaC proteins of Escherichia coli and Salmonella typhimurium by cloning into a high copy number plasmid. PMID- 6292204 TI - The Escherichia coli dnaC gene product. II. Purification, physical properties, and role in replication. AB - The Escherichia coli dnaC protein, purified to homogeneity from overproducing plasmid strains, is a polypeptide of 31,000 daltons (determined on a denaturing gel). The native molecular weight as calculated from the sedimentation coefficient of 2.75 S and Stokes radius of 24.5 A is 29,000. dnaC protein is N ethylmaleimide sensitive (Wickner, S., Berkower, L., Wright, M., and Hurwitz J. (1973) Proc. Natl. Acad. Sci. U. S. A. 70, 2369-2373), and has 3 sulfhydryl groups as determined with [14C]p-chloromercuribenzoate. The activity was assayed by complementation of a mutant dnaC extract or by reconstitution of a purified protein system which converts phi X174 single-stranded DNA to the duplex replicative form. In this conversion the dnaC protein is required during the initial prepriming stage of phi X174 DNA replication. Antiserum against dnaC protein specifically inhibits this stage but not the subsequent priming and elongation steps carried out by primase and the PolIII holoenzyme. Requirement for dnaC protein was also manifested in the in vitro replication of a plasmid DNA containing the E. coli origin of replication (oriC) by complementation of a mutant extract and specific inhibition by dnaC antiserum. PMID- 6292205 TI - The Escherichia coli dnaC gene product. III. Properties of the dnaB-dnaC protein complex. AB - The Escherichia coli dnaB and dnaC proteins form a tight complex in the presence of ATP (Wickner, S., and Hurwitz, J., (1975) Proc. Natl. Acad. Sci. U. S. A. 72, 921-925). The complexed dnaC protein is resistant to inhibition by the sulfhydryl reagent, N-ethylmaleimide. This protection is not observed when ATP is substituted by AMP, ADP, adenyl 5'-yl imidodiphosphate, or adenosine-5'-O-(3 thiotriphosphate); dATP provides partial protection. A sedimentation coefficient of 15.2 S determined by glycerol gradient sedimentation and a Stokes radius of 64 A determined by gel filtration suggests a molecular weight in the range of 400,000. The complex isolated by DEAE-cellulose chromatography contains six dnaC protein monomers of 29,000 daltons contains six dnaC protein monomers of 29,000 daltons contains six dnaC protein monomers of 29,000 daltons per dnaB protein hexamer (300,000 daltons) consistent with a calculated weight of 474,000. The isolated dnaB-dnaC protein complex functions in vitro in the replication of phage phi X174 single-stranded DNA to the duplex replicative form. Tritium-labeled dnaC protein, absent from an isolated prepriming com-dnaC protein, absent from an isolated prepriming complex intermediate, was nevertheless bound to the phiX replicative form DNA synthesized in vitro. These results suggest that stable inclusion od dnaC protein in the priming complex bound to DNA requires a completely assembled primosome. PMID- 6292206 TI - Luteinizing hormone receptors and gonadotropic activation of purified rat Leydig cells. AB - Specific receptors and metabolic responses to luteinizing hormone (LH) were analyzed in testicular Leydig cells purified by centrifugation of collagenase dispersed rat interstitial cells on density gradients of 14-32% Metrizamide. This procedure separated the interstitial cells into an upper, poorly responsive fraction and a lower, more dense population with high LH receptor content and prominent cyclic AMP and testosterone responses of gonadotropic stimulation. The upper layer consisted of morphologically heterogeneous and extensively vacuolated cells, of which relatively few were structurally identifiable as Leydig cells. The lower layer was comprised of almost homogenous Leydig cells when analyzed by electron microscopy and autoradiography, and sedimented as two adjacent bands with similar hormonal responses and densities of 1.085 and 1.105 g/cm3. The lighter and less responsive cell population appeared to result from the presence of damaged cells in the interstitial cell preparation and could be removed during density gradient isolation of the homogeneous and biologically active Leydig cell fraction. The more dense and active Leydig cell population retained hormonal responsiveness during culture for 24 h and showed loss of LH receptors and low concentrations of gonadotropin in vitro. These findings emphasize the importance of appropriate fractionation of rat interstitial cells to isolate the structurally intact and functionally active population of Leydig cells during studies on LH receptors and section in the testis. PMID- 6292207 TI - Evidence that calmodulin is in the chloroplast of peas and serves a regulatory role in photosynthesis. PMID- 6292208 TI - Transformation-dependent alterations is glycoproteins of extracellular matrix of human fibroblasts. Characterization of GP250 and the collagen-like GP140. AB - The extracellular matrix, prepared by extraction of confluent cultures of human lung WI-38 fibroblasts with a dipolar tonic detergent, contains four major glycoproteins: fibronectin, GP250, GP170, and GP140. All the glycoproteins can be surface-labeled; however, only fibronectin and GP170 can be readily removed by digestion with trypsin (Carter, W. G., and Hakomori, S. (1981) J. Biol. Chem. 256, 6953-6960). Most of the noncovalently bound GP250, GP170, and GP190, an additional minor glycoprotein, can be dissociated from the matrix by extraction with 8 M urea. The remaining insoluble matrix is stabilized by extensive intermolecular disulfide bonds and contains primarily GP140 and fibronectin (Carter, W. G. (1982) J. Biol. Chem. 257, 3249-3257). Affinity-purified, monospecific antibodies were prepared against GP[140 and fibronectin and utilized for detection of GP140 and fibronectin in extracts and conditioned media of WI 38, WI-38 VA13, WI-26, WI-26 VA4, and HT-1080 cells. Additional affinity purified, polyspecific antibodies that react with GP250, GP190 GP170, and GP140 were also utilized. Fibronectin, GP250, GP190, GP170, and GP140 were all absent from transformed cells. With the exception of GP140, the absence of these glycoproteins from the matrix of transformed cells was paralleled by their accumulation in the conditioned culture media. Incubation of conditioned culture media with collagenase indicated that GP190, GP170, and GP140, as well as other glycoproteins, were digested. Antibodies to GP140 did not react with any other cellular component indicating that it is not a processing product of other matrix glycoproteins. GP140 has characteristics unlike all reported collagen types and appears to be a new collagen-like glycoprotein. In contrast, neither Gp250 nor fibronectin were sensitive to digestion with collagenase. Antibodies that react with GP250 did not react with fibronectin and vice versa, suggesting that GP250 and fibronectin do not share antigenic determinants. The interaction of labeled fibronectin and the labeled, gelatin-binding domain of fibronectin with cells after fractionation on polyacrylamide gels indicated that GP170 is the primary procollagen receptor for fibronectin in the extracellular matrix. GP140 also bound fibronectin but to a lesser degree. Soluble GP170 and GP190 present in the conditioned medium of cultured cells also bound to insolubilized fronectin, confirming the association of GP170 and GP190 with fibronectin. The interaction of the glycoprotein components in the matrix are discussed in relation to their potential cooperative function in cell attachment and their failure to adhere to the surface of transformed cells. PMID- 6292209 TI - Copy number of a human type I alpha 2 collagen gene. AB - HpCl, a Charon 4A bacteriophage containing a 16.3-kilobase insert of human genomic DNA, has been identified as representing the 3' portion of the human alpha 2(I) collagen gene by hybrid selected translation and cross-reactivity with characterized sheep and chick collagen genes. Evaluation of HpCl by restriction endonuclease mapping and alpha 2(I) mRNA hybridization demonstrated that the alpha 2(I) mRNA coding regions are separated by noncoding regions including one of 2.7 kilobase pairs near the 3' end of the mRNA coding region. In addition, a 1.8-kilobase XbaI-BamHI fragment containing the 2' a 1.8-kilobase XbaI-BamHI fragment containing the 3' end of the alpha 2(I) mRNA coding region includes a region of DNA that is repeated many times throughout the human genome. Quantification of alpha 2(I) gene number, using the technique of DNA dot hybridization, with two EcoRI fragments from HpClo as probes, demonstrated that HpCl is represented once in the human genome. In this context, it is unlikely that human alpha 2(I) collagen chain production is modulated by differential transcription of multiple alpha 2(I) genes. PMID- 6292210 TI - The nucleotide sequence of the replication origin beta of the plasmid R6K. AB - We h ave identified by molecular cloning a region of 283 base pairs of the HindIII 2 fragment of R6K which corresponds to the region of the replication origin beta. This 283 base-pair DNA fragment, when present contiguously with the structural gene for the replication initiation protein of R6K, encoded in the HindIII 9-15 and part of HindIII 2 restriction fragments, will support the replication of a plasmid chimera containing the pBR322 replicon in a pol Ats host at the nonpermissive temperature. The nucleotide sequence of the region of replication origin beta has been determined. The nucleotide sequence has some homology with the ori gamma region of R6K; it has a 15-base-pair homology with the replication origin of Escherichia coli. PMID- 6292211 TI - Isolation of plasma membrane vesicles from rabbit skeletal muscle and their use in ion transport studies. AB - A method has been developed for the isolation of sealed plasma membrane vesicles from rabbit white skeletal muscle. The final preparation was highly purified as indicated by enrichment of plasma membrane marker enzymes (i.e. ouabain-sensitive (Na+,K+)-ATPase, adenylate cyclase, and acetylcholinesterase). The absence of sarcoplasmic reticulum and mitochondria as contaminants was indicated by the low specific activity of marker enzymes, i.e. Ca2+-ATPase, succinate-cytochrome c reductase, and monoamine oxidase. Thin section and negative staining electron microscopy confirmed the absence of sarcoplasmic reticulum and mitochondrial contamination. The plasma membrane preparation consisted largely of sealed vesicles as observed by electron microscopy and as also demonstrated by latency of enzymic activities, which were unmasked by preincubation with detergent (sodium dodecyl sulfate). Membrane sidedness was estimated from latency of ouabain-sensitive (Na+,K+)-ATPase activity and acetylcholinesterase activity. The latency studies suggest that most of the vesicles are oriented inside out with respect to the orientation of the sarcolemma membrane in the muscle fiber. The inside-out plasma membrane vesicles actively accumulated sodium ions upon addition of ATP. The sodium ions were concentrated greater than 8-fold inside the vesicles and were released upon addition of the ionophore monensin. The sodium ions were taken up in the presence of K+ or NH4+ but not of choline. Uptake was inhibited by low concentrations of vanadate or digitoxin. The Na+ uptake was concomitant with Rb+ efflux. Therefore, the sodium ion transport and the resulting gradients formed appear to have been generated by the ouabain-sensitive (Na+,K+)-ATPase. Batrachotoxin, which opens Na+ channels in excitable tissues, prevents most of the Na+ uptake, suggesting the presence of toxin-activated Na+ channels in these plasma membrane vesicles. PMID- 6292212 TI - A lymphoma protein with an in vitro site of tyrosine phosphorylation homologous to that in pp60src. AB - The major in vitro substrate for a tyrosine protein kinase in the particulate fraction of the lymphoma cell line LSTRA is a protein of molecular weight of 58,000 (pp58) (Casnellie, J. E. Harrison, M. L., Pike, L. J., Hellstrom, K. E., and Krebs, E. G. (1982) Proc. Natl. Acad. Sci. U. S. A. 79, 282-286). In order to determine if this protein was related to pp60src, the transformation-specific protein from Rous sarcoma virus, partial proteolysis maps of in vitro 32P-labeled pp58 and pp60src were prepared using Staphylococcus aureus V8 protease and papain. The maps were clearly different, indicating the pp58 is distinct from pp60src. However characterization of the tryptic fragment containing the single site of in vitro tyrosine phosphorylation in pp58 has revealed that the amino acid sequence around this site is extremely homologous to, if not identical with the sequence around the site of tyrosine phosphorylation in pp60src. PMID- 6292213 TI - Interferon receptors. Cross-linking of human leukocyte interferon alpha-2 to its receptor on human cells. PMID- 6292214 TI - The saxitoxin receptor of the sodium channel from rat brain. Evidence for two nonidentical beta subunits. AB - The saxitoxin receptor of the sodium channel purified from rat bran contains three types of subunits: alpha with Mr approximately 270,000, beta 1 with Mr approximately 39,000, and beta 2 with Mr approximately 37,000. These are the only polypeptides which quantitatively co-migrate with the purified saxitoxin receptor during velocity sedimentation through sucrose gradients. beta 1 and beta 2 are often poorly resolved by gel electrophoresis in sodium dodecyl sulfate (SDS), but analysis of the effect of beta-mercaptoethanol on the migration is covalently attached to the alpha subunit by disulfide bonds while the beta 1 subunit is not. The alpha and beta subunits of the sodium channel were covalently labeled in situ in synaptosomes using a photoreactive derivative of scorpion toxin. Treatment of SDS-solubilized synaptosomes with beta-mercaptoethanol decreases the apparent molecular weight of the alpha subunit band without change in the amount of 125I labeled scorpion toxin associated with either the alpha or beta subunit bands. These results indicate that the alpha and beta 1 subunits are labeled by scorpion toxin whereas beta 1 is not and that the beta 2 subunit is covalently attached to alpha by disulfide bonds in situ as well as in purified preparations. PMID- 6292215 TI - Leukotriene A4: preparation and enzymatic conversion in a cell-free system to leukotriene B4. AB - Treatment of leukotriene A4 (LTA4) methyl ester with sodium hydroxide in aqueous methanol at 4 degrees C afforded LTA4, the presence of which was inferred from the UV spectrum of the compound, its rate of reaction with water, and the identity of the hydration products obtained. The half-life of LTA4 in water (pH 7.4, room temperature) was increased from 14 to 500 s by 1 mg/ml of bovine serum albumin. This stabilized (chiral) LTA4 was converted to LTB4 by an epoxide hydrolase activity in the 100,000 x g supernatant fraction from sonified rat basophilic leukemia cells. Neither the ester of LTA4 nor the biologically incorrect enantiomer of LTA4 was metabolized to LTB4 under these conditions. PMID- 6292216 TI - Immunochemical analysis of the turnover of ubiquitin-protein conjugates in intact cells. Relationship to the breakdown of abnormal proteins. AB - Previous studies in a cell-free proteolytic system from reticulocytes indicated that the conjugation of ubiquitin with proteins plays a role in protein breakdown. To examine some of the physiological functions of the ubiquitin conjugation system, and immunochemical method was developed for the isolation of ubiquitin-protein conjugates from intact cells. A specific antiserum was raised against ubiquitin and purified by affinity chromatography on ubiquitin-Sepharose. When cells are labeled with tryptophan (which is missing from ubiquitin), labeled immunoreactive material isolated by the antibody is derived from the protein moiety of ubiquitin-protein conjugates. There is a marked increase in the labeling of ubiquitin-protein conjugates during the formation of abnormal proteins in reticulocytes (induced by the incorporation of amino acid analogs), suggesting that proteins with abnormal structure are more readily conjugated to ubiquitin than most normal proteins. Essentially similar, although less marked, effects of amino acid analogs were observed in Ehrlich ascites cells. When further protein synthesis was blocked with cycloheximide, ubiquitin conjugates decayed more extensively than the corresponding average labeled cellular proteins. This is consistent with the interpretation that a considerable part of ubiquitin conjugates is derived from a pool of rapidly degradable proteins. PMID- 6292217 TI - Glycosylation and intracellular transport of membrane glycoproteins encoded by murine leukemia viruses. Inhibition by amino acid analogues and by tunicamycin. AB - Addition of asparagine-linked oligosaccharides to nascent murine leukemia virus (MuLV)-encoded membrane glycoproteins was inhibited either completely by tunicamycin or specifically at Asn-X-Thr glycosylation sites by incorporation of the threonine analogue beta-hydroxynorvaline. In conditions of partial analogue substitution, a series of subglycosylated components is formed which are related by a constant apparent Mr difference when assayed by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. The total number of asparagine-linked oligosaccharides is then estimated by dividing the measured apparent Mr of one oligosaccharide into the total apparent Mr difference between the complete glycoprotein and the polypeptide chain that is synthesized in cells incubated with tunicamycin. Correct results were obtained using glycoproteins with known numbers of oligosaccharides. Our analyses indicate that the gp70 membrane envelope glycoproteins of certain ecotropic MuLVs contain seven oligosaccharides, whereas the GIX+ antigen-containing variant gp70 contains one fewer Asn-X-Thr-linked oligosaccharide. The membrane glycoprotein encoded by the gag gene of Friend MuLV contains only one asparagine-linked oligosaccharide. Similarly, the gp55 membrane glycoprotein encoded by Friend erythroleukemia virus contains four asparagine-linked oligosaccharides. Pulse-chase and cell surface iodination analyses indicate that MuLV membrane envelope glycoprotein processing by partial proteolysis and transport to the cell surface can be efficiently blocked by structural perturbations caused by incorporation of different amino acid analogues or by loss of oligosaccharides. Our data also suggest that loss of oligosaccharides may expose new antigenic sites in viral membrane glycoproteins and increase their susceptibility to intracellular proteolysis. PMID- 6292218 TI - Characterization of the rabbit renal receptor for native parathyroid hormone employing a radioligand purified by reversed-phase liquid chromatography. AB - Iodinated native bovine parathyroid hormone (bPTH(1-84)) was separated from uniodinated hormone by reversed-phase liquid chromatography techniques after lactoperoxidase labeling. Analysis of iodinated residues after enzymatic digestion indicated that the major labeled product was largely monoiodinated on the sole tyrosine residue. This material retained full bioactivity in an in vitro renal adenylate cyclase assay. Binding of 125I-bPTH(1-84) to rabbit renal membranes at 4 degrees C was proportional to membrane protein concentration and was saturable and dissociable. Radioligand binding was inhibited by concentrations of unlabeled bPTH(1-84) required to stimulate adenylate cyclase in the same membrane preparation but was not inhibited by non-PTH peptides other than adrenocorticotropin at high concentrations (greater than 10 microM). Synthetic NH2-terminal analogues of bPTH(1-84) all elicited approximately equivalent inhibition of radioligand binding which was, however, less potent than unlabeled bPTH(1-84), suggesting a role for the carboxyl region of the molecule in the interaction of bPTH(1-84) with its receptor. Activity of the NH2-terminal agonists was similar to bPTH(1-84) in stimulating adenylate cyclase. Although substitution in sequence position one, of serine in human PTH(1-34) for alanine in bPTH(1-34), reduced activity in the adenylate cyclase assay, inhibition of 125I-bPTH(1-84) binding by both peptides and by an analogue of bPTH(3-34) was equivalent, consistent with a minimal contribution of the first 2 residues for receptor binding of the NH2-terminal region of PTH. The results illustrate the utility of the radiolabeled preparation of native bPTH we have developed and emphasize the importance of probing the PTH receptor with an intact hormone to maximize information concerning the mechanism of PTH action. PMID- 6292219 TI - Selective solubilization of hyaluronic acid from fibroblast substratum adhesion sites. PMID- 6292220 TI - Lipoprotein regulation of cholesterol metabolism in macrophages derived from human monocytes. PMID- 6292221 TI - Characterization of cyclic AMP-requiring yeast mutants altered in the regulatory subunit of protein kinase. AB - The CYR3 mutant of yeast, Saccharomyces cerevisiae, partially accumulated unbudded cells and required cAMP for the best growth at 35 degrees C. The CYR3 mutation was partially dominant over the wild type counterpart and suppressed by the bcy1 mutation which is responsible for the deficiency of the regulatory subunit of cAMP-dependent protein kinase. The molecular weights of cAMP-dependent protein kinase and its catalytic and regulatory subunits were 160,000, 30,000, and 50,000, respectively. No significant differences in the molecular weights of cAMP-dependent protein kinase and the subunits were found between the wild type and CYR3 mutant strains. However, the cAMP-dependent protein kinase activity of CYR3 cells showed significantly higher Ka values for activation by cAMP at 35 degrees C than those of wild type and a clear difference in the electrophoretic mobility of the regulatory subunit was found between the wild type and CYR3 enzymes. The CYR3 mutation was suppressed by the IAC mutation which caused the production of a significantly high level of cAMP. The results indicate that the CYR3 phenotype was produced by a structural mutation in the CYR3 gene coding for the regulatory subunit of cAMP-dependent protein kinase in yeast. PMID- 6292222 TI - Regulation of rat liver cytosolic CTP: phosphocholine cytidylyltransferase by phosphorylation and dephosphorylation. PMID- 6292223 TI - Crystal structure and electron transfer properties of cytochrome c3. AB - The crystal structure of cytochrome c3 from the sulfate-reducing bacteria Desulfovibrio desulfuricans, Norway strain, has been determined through the fitting of the recently completed primary structure to a 2.5 A resolution electron density map. The phase calculations were based on three mercurial derivatives; anomalous scattering data were used to refine the four heme iron positions. A preliminary refinement of the molecular model has led to a conventional crystallographic R factor of 34%. Cytochrome c3 is folded in two structural domains with one heme in each, the two other heme moieties lying in a large groove dividing the molecule. The core of the protein is the compact four heme cluster which presents a relatively high degree of solvent exposure. The structural pattern of redox centers suggests that electron transfer might occur through direct contacts between some of the heme groups, via the overlapping system of pi oribitals or via intervening amino acid side chains or both. PMID- 6292224 TI - Induction of the messenger ribonucleic acid coding for phosphoenolpyruvate carboxykinase in H4-II-E cells. Evidence for a nuclear effect of cyclic AMP. AB - The effect of N6,O2'-dibutyryl cyclic adenosine monophosphate (Bt2cAMP) on the induction of the mRNA coding for the enzyme phosphoenolpyruvate carboxykinase was examined in H4-II-E cells. this mRNA comprised about 0.1% of total cellular poly(A)+RNA activity in uninduced cells and was increased 5- to 7-fold by the cyclic nucleotide. The maximal level was reached 3 h after addition of the nucleotide to the cell culture. This induction is attributed to cAMP since the nonmetabolizable analogs 8-bromocAMP and 8-(4-chlorophenylthio)cAMP produce inductions comparable to Bt2cAMP while sodium butyrate and dibutyryl cyclic GMP had little effect. The increased translational activity correlated well with a proportionate increase in the amount of phosphoenolpyruvate carboxykinase (P enolpyruvate carboxykinase) mRNA sequences which were hybridizable to a specific cDNA probe. Blot hybridization of total nuclear RNA isolated from uninduced H4-II E cells revealed eight P-enolpyruvate carboxykinase RNA sequence species ranging in size from 1.8 to 6.9 kilobases. Treatment with Bt2cAMP increased the amount of all eight of these forms. This increase became maximal by 45-60 min and was maintained for at least 1 h. In contrast, analysis of cytoplasmic RNA showed a single 3.2-kilobase (23 S) band, which was still increasing in amount 2 h after Bt2cAMP treatment. Thus, Bt2cAMP resulted in a sequential induction of nuclear P enolpyruvate carboxykinase RNA sequences followed by an increase in cytoplasmic phosphoenolpyruvate carboxykinase mRNA. We conclude that cyclic AMP exerts its main effect on P-enolpyruvate carboxykinase induction at the nuclear level. PMID- 6292225 TI - Characterization and biological properties of chemically deglycosylated human chorionic gonadotropin. Role of carbohydrate moieties in adenylate cyclase activation. PMID- 6292226 TI - Sugar transport by the bacterial phosphotransferase system. Radioactive and electron paramagnetic resonance labeling of the Salmonella typhimurium phosphocarrier protein (HPr) at the NH2-terminal methionine. PMID- 6292227 TI - Sugar transport by the bacterial phosphotransferase system. The glucose receptors of the Salmonella typhimurium phosphotransferase system. AB - We have previously reported that glucose can be phosphorylated by phospho-HPr and two sugar-specific pairs of proteins of the Escherichia coli and Salmonella typhimurium phosphoenolpyruvate:glycose phosphotransferase system. Each of the sugar-specific complexes comprises two proteins, lipid, and divalent cation, and each is present in membranes isolated from wild type cells. For reasons described in this report, one of the complexes is designated IIGlc and the other IIMan. The IIMan complex has previously been separated into its protein components, II-A and II-B (Kundig, W., and Roseman, S. (1971) J. Biol. Chem. 246, 1407-1418), while the accompanying reports describe dissociation of the IIGlc complex into its components, IIIGlc and II-BGlc. Curtis and Epstein (Curtis, S. J., and Epstein, W. (1975) J. Bacteriol. 122, 1189-1199) first showed that there are two phosphotransferase systems in whole cells responsible for glucose uptake and obtained the respective mutants, now designated ptsG and ptsM. The present studies provide kinetic conditions for assaying each activity separately (in vivo and in vitro), when both are present in the same membrane preparation. The IIGlc system is responsible for the uptake and phosphorylation of glucose and methyl alpha-glucoside, whereas the IIMan system is less specific and utilizes glucose, mannose, and 2-deoxyglucose. With high sugar concentrations in vitro, IIMan is also capable of phosphorylating methyl alpha-glucoside, fructose, and N acetylmannosamine, while IIGlc phosphorylates fructose and mannose. The in vivo transport results were qualitatively consistent with the in vitro phosphorylation results, and several of the kinetic parameters also showed good quantitative agreement. The levels of the two activities depended on the growth conditions. In addition, transport studies showed that initial uptake rates of methyl alpha glucoside and steady state levels of this analogue depended on the energy state of the cells and that these two parameters did not necessarily change in the same direction when metabolic inhibitors were used. A series of E. coli and S. typhimurium mutants were characterized both with respect to their ability to transport the glucose analogues and to phosphorylate them in vitro. The original mutants of Curtis and Epstein, ptsG and ptsM, were found to be defective in II BGlc and the IIMan complex, respectively. PMID- 6292228 TI - Characterization of the adrenoreceptor activities of isoprenaline in the field stimulated rat vas deferens: selective supersensitivity to beta 2-mediated responses following reserpine treatment. AB - 1 Low concentrations of isoprenaline (EC50 = 45.6 nM) inhibited contractions in the isolated field stimulated rat vas deferens. This inhibitory effect was markedly attenuated by the postjunctional beta 2-adrenoreceptor antagonist timolol, but not affected by the prejunctional alpha 2 or postjunctional alpha 1 adrenoreceptor antagonists rauwolscine and prazosin, respectively. 2 In vas deferens of rats previously treated with reserpine, the postjunctional beta 2 adrenoreceptor-mediated inhibitory response to isoprenaline was markedly potentiated. 3 High concentrations of isoprenaline (EC50 = 1.5 microM) also inhibited contractility in tissues in which postjunctional beta 2-adrenoreceptors were maximally blocked by high concentrations of timolol. This contractile inhibition produced by isoprenaline was abolished by rauwolscine but not significantly altered by prazosin or pretreatment of the rats with reserpine indicating stimulation of prejunctional alpha 2-adrenoreceptors. 4 Rauwolscine pretreatment unmasked an ability of isoprenaline (EC50 = 17.1 microM) to produce enhancement of field stimulation-induced contractions. This response was abolished by prazosin but was unaffected by timolol or reserpinization indicating an action upon postjunctional alpha 1-adrenoreceptors. 5 The data indicate isoprenaline activates adrenoreceptor mechanisms in the field stimulated rat vas deferens by a direct action not dependent upon endogenous catecholamines and with an order of activity of beta 2 much greater than alpha 2 greater than alpha 1. Pretreatment with reserpine produces rapid and selective development of supersensitivity to the postjunctional beta 2-mediated inhibitory response of isoprenaline in this preparation. PMID- 6292229 TI - Molecular and macroscopic properties of PMMA bone cement: free-radical generation and temperature change versus mixing ratio. AB - The molecular and macroscopic changes occurring during the polymerization of poly(methyl methacrylate) (PMMA) bone cement have been investigated. Electron paramagnetic resonance (EPR) spectroscopy was used to monitor free-radical generation and this was compared to temperature changes occurring in the cement for various ratios of polymer powder to liquid monomer (P/L ratio) used in the sample preparation. Both the concentration and the characteristic growth time of the free radicals associated with the polymerization of the bone cement depended on the P/L ratio used. Larger P/L ratio resulted in shorter characteristic growth time for the free radicals as well as a shorter time for the occurrence of the peak sample temperature. Smaller P/L ratios gave smaller maximum concentrations of free radicals and larger peak temperatures. These results are explained on the basis of (1) more initiators present at higher P/L ratios resulting in faster polymerization and (2) less initiators and more monomers present at smaller P/L ratios resulting in fewer radicals but more exothermic reactions. The free radicals present in the bone cement due to the manufacturer's sterilization process were found to be proportional to the fraction of powder used in the preparation, indicating negligible monomer loss during sample mixing. PMID- 6292230 TI - In vivo surface activity of a hydroxyapatite alveolar bone substitute. PMID- 6292231 TI - Chronic erosive gastritis--clinical and radiological features. AB - Chronic erosive gastritis has been regarded as an unusual, ill-defined condition of unknown etiology. We report a retrospective study of 40 patients in whom this diagnosis was made relating radiologic findings to patients' age, symptomatology and possible etiologic factors. The condition is more common than previously suspected and requires careful technique combining double contrast and graded compression to demonstrate fine mucosal abnormalities. The radiologic appearance is of mucosal mounds measuring 3-11 mm in diameter with central barium collections and numbering from 3-27 per examination. Antral involvement occurred in 90% of patients, with diffuse involvement of both antrum and body in 58%. Several linear erosions measuring up to 15 mm in length were also demonstrated. Chronic erosive gastritis is most severe and symptomatic between 40 and 60 years, becoming sub-clinical in the elderly. Symptoms mimic those of a peptic ulcer or gastric neoplasm and are more severe the more extensive the disease. Gastric irritants, virus infection and Crohn's disease were possible etiologic factors in some patients in this study. PMID- 6292232 TI - Characterization of primary rabbit kidney cultures that express proximal tubule functions in a hormonally defined medium. AB - Primary cultures of rabbit-kidney epithelial cells derived from purified proximal tubules were maintained without fibroblast overgrowth in a hormone-supplemented serum-free medium (Medium RK-1). A hormone-deletion study indicated that the primary cultures derived from purified rabbit proximal tubules required all of the three supplements in Medium RK-1 (insulin, transferrin, and hydrocortisone) for optimal growth but did not grow in response to EGF and T3. In contrast, the epithelial cells in primary cultures derived from an unpurified preparation of rabbit kidney tubules and glomeruli grew in response to EGF and T3, as well as insulin, transferrin, and hydrocortisone. These observations suggest that kidney epithelial cells derived from different segments of the nephron grow differently in response to hormones and growth factors. Differentiated functions of the primary cultures derived from proximal tubules were examined. Multicellular domes were observed, indicative of transepithelial solute transport by the monolayers. The proximal tubule cultures also accumulated alpha-methylglucoside (alpha-MG) against a concentration gradient. However, little or no alpha-MG accumulation was observed in the absence of Na+. Metabolic inhibitor studies also indicated that alpha-MG uptake by the primaries is an energy-dependent process, and depends upon the activity of the Na+/K+ ATPase. Phlorizin at 0.1 mM significantly inhibited 1 mM alpha-MG uptake whereas 0.1 mM phloretin did not have a significant inhibitory effect. Similar observations have been made concerning the Na+-dependent sugar transport system located on the lumenal side of the proximal tubule, whereas the Na+-independent sugar transporter on the peritubular side is more sensitive to inhibition by phloretin than phlorizin. The cultures also exhibited PTH-sensitive cyclic AMP synthesis and brush-border enzymes typical of proximal cells. However, the activities of the enzymes leucine aminopeptidase, alkaline phosphatase, and gamma-glutamyl-transpeptidase were lower in the cultures than in purified proximal-tubule preparations from which they are derived. PMID- 6292233 TI - Effect of barium and tetraethylammonium on membrane circulation in frog retinal photoreceptors. AB - We studied the influence of altered ionic conditions on the recycling of synaptic vesicle membrane in frog retinal photoreceptors using horseradish peroxidase to monitor synaptic activity and trace the fate of internalized membrane. The addition of 1.2 mM barium or 20 mM tetraethylammonium to isolated retinas maintained in Ringer's solution, changes the usual balance of membrane circulation in the rod cells; the cone cells are much less affected. Retrieval of synaptic vesicle membrane in the rods, which normally regenerates small vesicles, becomes mediated predominantly by large sacs and vacuoles ("cisternae"). Because these cisternae can be labeled with peroxidase, they appear to arise from endocytized membrane. Morphometric analysis suggests strongly that the cisternae are formed of circulating synaptic vesicle membrane. The effects of barium and tetraethylammonium can be inhibited by high extracellular potassium, by high intensity light, and by 5 mM cobalt. They seem likely to depend on potassium channels, though additional more complex mediation may also be involved. The alterations in membrane retrieval that we find are of interest in terms of the multiple pathways of membrane cycling now being uncovered. They open potential experimental approaches to the controls of this circulation. In addition, the findings extend our previous ones demonstrating that rod cells and cone cells differ in their responses to divalent cations in ways that seem likely to be of physiological importance. PMID- 6292234 TI - Free and polymerized tubulin in cultured bone cells and Chinese hamster ovary cells: the influence of cold and hormones. AB - Free and polymerized tubulin were measured in bone cells and Chinese hamster ovary (CHO) cells cultured on plastic substrata. Polymerized tubulin was stabilized in a microtubule- stabilizing medium (MSM) containing 50 percent glycerol and separated from free tubulin by centrifugation. Tubulin content was assayed in both fractions by the colchicines- binding assay. The measured degree of polymerization in both bone cells and CHO cells varied with stabilixation conditions. The degree of polymerization in both bone cells and CHO cells varied with stabilization conditions. The degree of polymerization in both bone cells and CHO cells varied with stabilization conditions. The degree of polymerization in attached cells was found to increase up to 73 percent during the first 20 min after addition of the MSM at 24 degrees C, and remained constant thereafter. Stabilization of 0 degrees C resulted in a decrease down to 62 percent in the degree of constant thereafter. Stabilization at 0 degrees C resulted in a decrease down to 62 percent in the degree of polymerization during the first 20 min after addition of the MSM at 24 degrees C, and remained constant thereafter. Confluent bone cells maintained at 0 degrees C for 1 h before stabilization contained significantly less polymerized tubulin than control cells kept at 37 degrees C using stabilization both at 0 degrees C and at 24 degrees C. Changes in bone cell morphology induced by incubation of cells with prostaglandin E(1) or E(2), parthyroid hormone, and dibutyryl cyclic AMP were not associated with a change in the degree of tubulin polymerization. This was confirmed morphologically by immunofluorescence using affinity-purified tubulin antibodies: microtubules in hormone- treated cells were not noticeably reorganized when compared to microtubule organization in control cells. They were, however, squeezed closer together in cellular pseudopods due to the altered cell shape. This altered cell shape appears to be correlated with disorganization of the microfilament system, since microfilaments, detected using affinity-purified actin antibodies, did alter drastically their appearance and distribution after hormone addition. PMID- 6292235 TI - Protein translocation across the endoplasmic reticulum. I. Detection in the microsomal membrane of a receptor for the signal recognition particle. AB - Salt-extracted microsomal membranes (K-RM) contain an activity that is capable of releasing the signal recognition particle (SRP)-mediated elongation arrest of the synthesis of secretory polypeptides (Walter, P., and G. Blobel, 1981, J. Cell Biol., 91:557-561). This arrest-releasing activity was shown to be a function of an integral microsomal membrane protein, termed the SRP receptor (Gilmore, R., P. Walter, and G. Blobel, 1982, J. Cell Biol., 95:470-477). We attempted to solubilize the arrest-releasing activity of the SRP receptor by mild protease digestion of K-RM using either trypsin or elastase. We found, however, that neither a trypsin, nor an elastase "solubilized" supernatant fraction exhibited the arrest-releasing activity. Only when either the trypsin- or elastase-derived supernatant fraction was combined with the trypsinized membrane fraction, which by itself was also inactive, was the arrest-releasing activity restored. Release of the elongation arrest was followed by the translocation of the secretory protein across the microsomal membrane and the removal of the signal peptide. Thus, although we have been unable to proteolytically sever the arrest-releasing activity from K-RM and thereby to uncouple the release of the elongation arrest from the process of chain translocation, we have been able to proteolytically dissect and reconstitute the arrest-releasing activity. Furthermore, we found that the arrest-releasing activity of the SRP receptor can be inactivated by alkylation of K-RM with N-ethylmaleimide. PMID- 6292236 TI - Protein translocation across the endoplasmic reticulum. II. Isolation and characterization of the signal recognition particle receptor. AB - The signal recognition particle (SRP)-mediated elongation arrest of the synthesis of nascent secretory proteins can be released by salt-extracted rough microsomal membranes (Walter, P., and G. Blobel, 1981, J. Cell Biol, 91:557-561). Both the arrest-releasing activity and the signal peptidase activity were solubilized from rough microsomal membranes using the nonionic detergent Nikkol in conjunction with 250 mM KOAc. Chromatography of this extract on SRP-Sepharose separated the arrest-releasing activity from the signal peptidase activity. Further purification of the arrest-releasing activity using sucrose gradient centrifugation allowed the identification of a 72,000-dalton polypeptide as the protein responsible for the activity. Based upon its affinity for SRP, we refer to the 72,000-dalton protein as the SRP receptor. A 60,000-dalton protein fragment (Meyer, D. I., and B. Dobberstein, 1980, J. Cell Biol., 87:503-508) that had been shown previously to reconstitute the translocation activity of protease digested membranes, was shown here by peptide mapping and immunological criteria to be derived from the SRP receptor. Findings that are in part similar, and in part different from these reported here and in our preceding paper were made independently (Meyer, D. I., E. Krause, and B. Dobberstein, 1982, Nature (Lond.). 297:647-650) and the term "docking protein" was proposed for the SRP receptor. A lower membrane content of both SRP and the SRP receptor than that of membrane bound ribosomes suggests that the SRP-SRP receptor interaction may exist transiently during the formation of a ribosome-membrane junction and during translocation. PMID- 6292237 TI - Surfaces of rod photoreceptor disk membranes: light-activated enzymes. AB - The light-activated GTP-binding protein (GBP) in toad rod outer segments has been located on the cytoplasmic surface (CS) of rod disk membranes by correlating biochemical results with images of quick-frozen, freeze-fractured, and deep etched rod outer segments. This has been accomplished by selectively removing and replacing the 8-12-nm particles that are found on the CS of disk membranes, exactly in parallel with the GBP. In contrast, the large particles are not correlated with another major disk enzyme, the light-activated cGMP phosphodiesterase. We have been unable to visualize this protein. The surface density of large particles, one particle per eleven rhodopsins in isolated rod outer segments and one particle per nine rhodopsins in intact retina, correlates well with previous biochemical estimates of GBP numbers based on enzyme activity. After the identification of the large particles, we tested the effects of light on the density of particles on the surface of disk membranes in intact retinas. Retinas quick-frozen at various intervals after a bright flash of light show a modest increase (approximately 30%) in particle density by 10 s after the flash but no increase before 1 s. The number of particles on the disk membrane returns to dark levels between 1 and 10 min after the flash. The 1-s latency in the change of particle binding would appear to rule out this process as a mechanism for initiating phototransduction in the rod. PMID- 6292238 TI - Sites of inhibition of mitochondrial electron transport in macrophage-injured neoplastic cells. AB - Previous work has shown that injury of neoplastic cells by cytotoxic macrophages (CM) in cell culture is accompanied by inhibition of mitochondrial respiration. We have investigated the nature of this inhibition by studying mitochondrial respiration in CM-injured leukemia L1210 cells permeabilized with digitonin. CM induced injury affects the mitochondrial respiratory chain proper. Complex I (NADH-coenzyme Q reductase) and complex II (succinate-coenzyme Q reductase) are markedly inhibited. In addition a minor inhibition of cytochrome oxidase was found. Electron transport from alpha-glycerophosphate through the respiratory chain to oxygen is unaffected and permeabilized CM-injured L1210 cells oxidizing this substrate exhibit acceptor control. However, glycerophosphate shuttle activity was found not to occur within CM-injured or uninjured L1210 cells in culture hence, alpha-glycerophosphate is apparently unavailable for mitochondrial oxidation in the intact cell. It is concluded that the failure of respiration of intact neoplastic cells injured by CM is caused by the nearly complete inhibition of complexes I and II of the mitochondrial electron transport chain. The time courses of CM-induced electron transport inhibition and arrest of L1210 cell division are examined and the possible relationship between these phenomena is discussed. PMID- 6292239 TI - Lysosomal enzyme oligosaccharide phosphorylation in mouse lymphoma cells: specificity and kinetics of binding to the mannose 6-phosphate receptor in vivo. AB - Phosphomannosyl residues on lysosomal enzymes serve as an essential component of the recognition marker necessary for binding to the mannose 6-phosphate (Man 6-P) receptor and translocation to lysosomes. The high mannose-type oligosaccharide units of lysosomal enzymes are phosphorylated by the following mechanism: N acetylglucosamine 1-phosphate is transferred to the 6 position of a mannose residue to form a phosphodiester; then N- acetylglucosamine is removed to expose a phosphomonoester. We examined the kinetics of this phosphorylation pathway in the murine lymphoma BW5147.3 cell line to determine the state of oligosaccharide phosphorylation at the time the newly synthesized lysosomal enzymes bind to the receptor. Cells were labeled with [2-(3)H]mannose for 20 min and then chased for various times up to 4 h. The binding of newly synthesized glycoproteins to the Man 6-P receptor was followed by eluting the bound ligand with Man 6-P. Receptor bound material was first detected at 30 min of chase and reached a maximum at 60 min of chase, at which time approximately 10 percent of the total phosphorylated oligosaccharides were associated with the receptor. During longer chase times, the total quantity of cellular phosphorylated oligosaccharides decreased with a half-time of 1.4 h, suggesting that the lysosomal enzymes had reached their destination and had been dephosphorylated. The structures of the phosphorylated aligosaccharides of the eluted ligand were then determined and compared with the phosphorylated oligosaccharides of molecules which were not bond to the receptor. The major phosphorylated oligosaccharide species present in the nonreceptor-bound material contained a single phosphosphodiester at all time examined. In contrast, receptor-bound oligosaccharides were greatly enriched in species possessing one and two phosphomonoesters. These results indicate that binding of newly synthesized lysosomal enzymes to the Man 6-P receptor occurs only after removal of the covering N- acetylglucosamine residues. PMID- 6292240 TI - Correlation between electrical activity and ACTH/beta-endorphin secretion in mouse pituitary tumor cells. AB - The electrical and secretory activities of mouse pituitary tumor cells (AtT-20/D 16v), which contain and release the ACTH/beta-endorphin family of peptides, were studied by means of intracellular recordings and radioimmunoassays. Injection of depolarizing current pulses evoked action potentials in all cells and the majority (82%) displayed spontaneous action potential activity. Action potentials were found to be calcium-dependent. Barium increased membrane resistance, action potential amplitude and duration, and release of ACTH and beta-endorphin immunoactivity. Isoproterenol increased both action potential frequency and hormone secretion. Raising the external calcium concentration increased the frequency and amplitude of the action potentials and stimulated secretion of ACTH and beta-endorphin immunoactivity. Thus, stimulation of secretory activity in AtT 20 cells was closely correlated with increased electrical activity. However, a complete blockade of action potential activity had no effect on basal hormone secretion in these cells. These results suggest that the mechanisms underlying stimulated hormone secretion are different from those responsible for basal secretory activity. It is proposed that the increased influx of calcium due to the increased action potential frequency initiates the stimulated release of hormone from these cells. PMID- 6292242 TI - Cerebral Na+,K+-ATPase activity during exposure to and recovery from acute ischemia. AB - This study documents the Na+,K+-ATPase activity as well as selected parameters of oxidative metabolism and electrophysiological function in rat brain exposed to ischemia produced by electrocautery of the vertebral arteries and reversible occlusion of the carotid arteries. During a 0.5-h ischemic exposure in which the electroencephalograph (EEG) was abolished and energy metabolism severly compromised the Na+,K+-ATPase showed a capability for enhanced activity (120-140% of control). On recirculation, the Na+,K+-ATPase activity showed a phasic pattern, which was characterized by normal values at 0.25-2 h, increased values (115-125% of control) at 3-24 h, and, finally, normal values at 72 h of recirculation, respectively. The maintenance of Na+,K+-ATPase integrity was correlated with a gradual return of EEG activity and virtually complete restitution of the cerebral energy state during the 72 h of recirculation. Measurements of thiobarbituric acid reactive material and water soluble antioxidant during ischemia and recirculation gave no evidence of the presence of significant free radical lipid peroxidation in this model. It is concluded that Na+,K+-ATPase and its associated membrane lipids are not irreversibly damaged by ischemia in which the tissue lactacidosis is limited to less than 20 mumol g-1. PMID- 6292241 TI - Modification of cell surface glycoprotein: addition of fucosyl residues during epidermal differentiation. AB - When cutaneous sections from the newborn rat were treated with alpha-fucosidase, Ulex europeus agglutinin I (UEA) binding to the cell surface of the differentiated cells in the epidermis was diminished and there was an appearance in these cell layers of binding by Bandeiraea simplicifolia I-B4 lectin (BS I B4), which normally is specific for the basal cells. A similar treatment with alpha-galactosidase resulted in a loss of BS I-B4 binding, but had no effect on UEA binding. Glycoproteins isolated from the membranes of epidermal cells showed a threefold increase in the ratio of binding to UEA versus BS I-B4 affinity columns as the proteins were derived from the more differentiated cell populations. These data suggest that alpha-fucosyl residues are added to the glycoproteins on the cell surfaces of differentiated cells, thus blocking alpha galactosyl residues and changing the lectin binding specificity as epidermal cells move out of the basal cell layer. PMID- 6292243 TI - [Syndrome of the posterior branches of spinal nerves. Anatomic, symptomatologic and therapeutic basis]. AB - The author describes the clinical findings of the posterior rami of the spinal nerve syndrome (R. Maigne) which innervates the skin from the vertex down to the coccyx. Moreover, he reports the cutaneous trophic changes which occur during the suffering of the anterior rami of the spinal nerve. He gives, for example, the sufferings of the T12 rami which are at the origin of the major diagnosis errors. PMID- 6292244 TI - High-performance liquid chromatography of fat-soluble vitamins: separation and identification of vitamins D2 and D3 and their isomers in food samples in the presence of vitamin A, vitamin E and carotene. AB - Vitamins D2 and D3 and their corresponding previtamins and provitamins were resolved by reversed-phase high-performance liquid chromatography using a ternary solvent system (acetonitrile-methanol-water) pumped according to a gradient elution programme. The D vitamins were also resolved in the presence of other lipid-soluble vitamins (A, E and K1) and carotene. The peaks were monitored with a UV-visible variable-wavelength detector and were detected at their maximum absorbance, resulting in maximum sensitivity. Lipid-soluble vitamins and carotene were resolved in extracts obtained from oils and butter, thus permitting their identification in a single chromatographic run. PMID- 6292245 TI - Determination of airborne 2,4-toluenediisocyanate vapors. AB - An accurate and convenient sampling and analytical method was developed for airborne 2,4-toluenediisocyanate (TDI). A chromophoric derivatizing reagent was used to convert the TDI to a stable urea derivative during collection for subsequent quantification by liquid chromatography. The overall accuracy (at the 95% confidence level) and precision of the method are +/- 7.9% and +/- 10.0% respectively under laboratory conditions. Long term stability was observed for both the reagent coated sorbent tube and the TDI urea derivative. A TDI concentration of 1 ppb can be detected by taking a 15-1 sample volume. PMID- 6292246 TI - Determination of vitamin D3 in cod-liver oil by high-performance liquid chromatography. AB - The present British Pharmacopoeia monograph for cod-liver oil requires a bioassay for the vitamin D3 content which is both time-consuming and complex. Alternative assays employing chromatographic procedures have been described but all these involve prior saponification of the oil. A selective extraction for vitamin D3 without the need for saponification is reported in this paper. The extraction utilizes only chromatographic assay using argentation on reversed-phase silica, with vitamin D2 as the internal standard. Reproducibility of injection gave a coefficient of variation of 0.6%, and repeatability of extraction for six samples gave a coefficient of variation of 6.8%. PMID- 6292247 TI - Rapid and reliable method for the analysis of nucleotide pools by reversed-phase high-performance liquid chromatography. AB - A rapid and reliable protocol for the simultaneous separation of ribo-, deoxyribo and cyclic nucleotides has been developed using high-performance liquid chromatography on a C18 microBondapak column and isocratic elution with ammonium phosphate buffer 0.2 m, pH 5.1). Resolution of deoxyribonucleotides has been confirmed by performing resolution before and after periodate oxidation. The general order of elution is ribonucleotides, deoxyribonucleotides and cyclic nucleotides. While periodate oxidation improved the clarity of separation of deoxyribonucleotides by eliminating ribonucleotides, incorporation of methanol in the eluent shortened the retention time of cyclic nucleotides. The application of this method to a complex biological system is reported. PMID- 6292248 TI - Comparison of an ELISA technique with quantal micro-neutralization test for serotyping of HSV-1 or HSV-2-infected patients. AB - An enzyme-linked immunosorbent assay (ELISA) using semi-purified herpes simplex antigens extracted from cell nuclei was employed for typing of 32 sera collected from patients infected with HSV-1 or HSV-2. The results were in agreement with those obtained by the quantal micro-neutralization test. Moreover, sera with antibodies which could not be detected by micro-neutralization test could be typed by the ELISA technique according to the HSV isolates. PMID- 6292249 TI - Determination of infectivity of transforming avian sarcoma virus and parotitis virus in fibrin-embedded monolayer cultures. AB - A sensitive and reproducible technique for titration of transforming avian sarcoma virus, Rous sarcoma virus (RSV), in chick embryo cell monolayers embedded between two layers of a solid fibrin coagulum was developed. The foci of RSV transformed fibrinolysis-exhibiting cells locally attacked the surrounding fibrin and were identifiable as plaque-like cell-free areas of retraction easily scored against the background of cells stained with neutral red. In comparison to the conventional assay in agar, the assay in fibrin proved more sensitive and less time-consuming. The titration technique in fibrin was also used for plaquing a cytopathogenic virus, parotitis virus, but the titration by conventional plaque assay in agar and the technique in fibrin yielded similar results in this case. PMID- 6292250 TI - Solid-phase enzyme immunoassay for rotavirus antigen: faecal protease activity as a reason for false-negative results. AB - Rabbits and guinea pigs were immunized with purified bovine rotavirus. Immunoglobulin G fractions of the resulting antisera were used in a standard four layer solid-phase enzyme immunoassay (EIA) for rotavirus antigen in human faecal specimens. Samples negative for rotavirus in electron microscopy, when diluted in standard EIA buffers, regularly gave absorbance values lower than those obtained with buffer blank only. By further diluting of the samples the resulting absorbance values were found to increase to the blank levels. When all dilution buffers were supplemented with 1-5% of bovine serum, negative samples at any dilution gave absorbance values close to those of the buffer blanks. Similar results were obtained if the serum was replaced by 1-5 mM of phenylmethylsulphonyl fluoride, a synthetic broad spectrum serine-type protease inhibitor. Aprotinin, another protease inhibitor, was without effect. A similar inhibition pattern was obtained when faecal specimens were tested in a caseinolytic quantitative protease assay in the presence of the above inhibitors. These observations suggest that protease activity present in human faecal samples may cause false-negative results in solid-phase immunoassay for viral antigens, unless appropriate means are used to avoid this interference. PMID- 6292251 TI - Quantification of an Epstein-Barr virus-associated membrane antigen component. AB - A method is described for the preparation of a 125I-labelled membrane antigen (MA) component (gp340) from B95-8 cell membranes using sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). Good yields of antigenic material were obtained when renaturation of the [125I]gp340 was carried out by removal of SDS in the presence of urea and subsequent removal of the urea. The availability of purified, radiolabelled gp340 has provided the essential basis for the development of a radioimmunoassay which, for the first time, permits quantification of this antigen. The assay has been used to demonstrate that cell membrane MA is a better source of gp340 for large-scale work than is the Epstein Barr virus envelope and to measure the increase in expression of gp340 following treatment of cells with 12-O-tetradecanoyl-phorbol-13-acetate (TPA). PMID- 6292252 TI - A profile of treatment approaches used at comprehensive cancer centers. AB - The Centralized Cancer Patient Data System is the system which the 21 comprehensive cancer centers in the U.S. have established in order to report and to analyze demographic, diagnostic, treatment and survival data on all new patients. We propose that this closely monitored standardized 36 item dataset can be used to profile the categories of initial treatment that are given to patients having all types and stages of cancer: the data may be displayed for all centers or used to compare the approaches used at different centers. Differences in the frequency with which surgery, radiation, chemotherapy, and no specific treatment are used for patients having any of three common histologic types of lung cancer serve to illustrate the method. Opportunities now exist to utilize this new resource to study trends in the use of treatment modalities and their relationship to survival, which in turn should enhance the accessibility of treatment information about all patients at cancer centers, rather than only those which are reported from selected treatment protocols. This approach may also be uniquely useful in obtaining treatment and survival information about patients with rare sites or types of cancer. PMID- 6292254 TI - Calcium potentiates the cyclic nucleotide and phosphaturic response to parathyroid hormone infusion. AB - To determine whether calcium modulates the action of PTH, we measured the cyclic nucleotide and phosphaturic response to PTH following a 4-h infusion of glucose (day 1) and calcium (day 2). The 12 subjects were selected to provide a range of low, normal, and high endogenous PTH function. PTH stimulated nephrogenous cAMP [185 +/- 31 nmol/100 ml glomerular filtrate (GF)], cyclic guanosine monophosphate (0.44 +/- 0.09 mumol/g creatinine), and phosphate (367 +/- 59 mg P/g creatinine) excretion. Calcium infusion stimulated nephrogenous cAMP excretion in the hypoparathyroid subjects (1.42 +/- 0.35 nmol/100 ml GF) but reduced it in subjects with normal parathyroid function (-2.22 +/- 0.46 nmol/100 ml GF). Calcium infusion stimulated cGMP (0.64 +/- 0.1 mumol/g creatinine) and phosphate (113 +/- 48 P/g creatinine) excretion in all subject groups. Calcium infusion led to a 2-fold increase in the cyclic nucleotide and phosphaturic response to PTH in the normal and hypoparathyroid subjects, but had little effect on the PTH response in hyperparathyroid subjects. The extent to which calcium potentiated the ability of PTH to stimulate nephrogenous cAMP excretion correlated negatively with the basal nephrogenous cAMP excretion (r = -0.685, P less than 0.01). We conclude that calcium potentiates the acute effects of PTH on renal cyclic nucleotide and phosphate excretion. This effect is modified by the basal levels of PTH stimulation of the kidney such that it is reduced in magnitude when basal PTH stimulation is increased. PMID- 6292253 TI - In vitro studies on prolactin release and adenylate cyclase activity in human prolactin-secreting pituitary adenomas. Different sensitivity of macro- and microadenomas to dopamine and vasoactive intestinal polypeptide. PMID- 6292255 TI - beta-Adrenoceptor-adenosine 3',5-monophosphate system in human leucocytes before and after treatment for hyperthyroidism. AB - Modifications in characteristics of the beta-adrenoceptor-cAMP system were observed in leucocytes from 10 patients in the hyperthyroid state after antithyroid treatment. These include 1) an increased number of beta-adrenoceptors without a change in their affinity, 2) an increased magnitude of stimulation of adenylate cyclase by isoprenaline, without a change in the NaF-stimulated enzyme activity, 3) an increased cAMP-dependent protein kinase activity ratio, and 4) an increased activity of cAMP-phosphodiesterase. Moreover, the plasma cAMP levels were markedly elevated during the hyperthyroid state. It is suggested that the above changes may in part constitute the molecular basis for the reputed catecholamine supersensitivity in the hyperthyroid state. PMID- 6292256 TI - Perturbations of negative feedback sensitivity in agonadal patients undergoing estrogen replacement therapy. AB - Urinary gonadotropin excretion was measured in 30 patients with gonadal dysgenesis, aged 2 months to 17 yr. Between bone ages 3-8 yr, mean FSH excretion (575 mIU/h) was elevated 8-fold in agonadal individuals compared to levels in intact prepubertal girls; mean urinary LH (49 mIU/h) in agonadal patients during this time period was increased nearly 2-fold over results from normal prepubertal females. Nine of 10 patients given 0.3 to 0.6 mg conjugated estrogen (Premarin) daily to initiate puberty exhibited prompt suppression of urinary gonadotropin levels from markedly elevated levels to within or very close to the normal prepubertal range. Such a response was found in only two of seven patients given 0.15 mg of the same drug. All instances of suppression were followed by escape from low levels of gonadotropin excretion as treatment was continued. Prior exposure to exogenous or endogenous estrogen markedly reduced the suppressive potential of treatment with 0.3 or 0.6 mg Premarin. A favorable advance of bone maturation in relation to chronological age was achieved by the administration of 0.15 mg Premarin daily, a dose which caused a satisfactory onset of secondary sex characteristics. IN CONCLUSION: 1) a component of gonadotropin restraint in midchildhood is supplied by the ovary; 2) adult castrate levels of gonadotropins are achieved in the agonadal patient of peripubertal age in the presence of a highly sensitive negative feedback axis between sex hormones and gonadotropins; 3) sex steroids themselves may modify the gonadotropin-gonadal negative feedback axis in patients with gonadal dysgenesis; and 4) puberty may be initiated favorably with conjugated estrogens in an oral dose of 0.15 mg daily. PMID- 6292257 TI - Suppression of adrenal mineralocorticoid production in prehypertensive young adult men. AB - We assessed the renin-aldosterone axis in response to treadmill exercise and adrenal steroidogenesis after graded ACTH infusions in two groups, each composed of nine young adult men from Bourbon County, Kentucky, from the upper and lower deciles of the blood pressure distribution. Those from the upper decile with relatively higher blood pressures, termed prehypertensive, had significantly lower plasma aldosterone and 18-hydroxycorticosterone concentrations after graded ACTH than those with relatively lower pressures, and had significantly lower urinary potassium excretions. Renin and aldosterone responses to exercise were also significantly blunted in the prehypertensive subjects. No differences were found in plasma cortisol, deoxycortisol, deoxycorticosterone, corticosterone, or dehydroepiandrosterone sulfate, although dehydroepiandrosterone was higher at one infusion rate in prehypertensives. These findings suggest that the adrenal mineralocorticoid pathway and the renin-aldosterone axis are suppressed in prehypertensive young males perhaps as an appropriate feedback response to their higher blood pressures. PMID- 6292258 TI - The regulation of urinary free 19-nor-deoxycorticosterone and its relation to systemic arterial blood pressure in normotensive and hypertensive subjects. AB - 19-Nor-deoxycorticosterone (19-nor-DOC) is a naturally occurring, potent mineralocorticoid present in hypertensive animal models as well as man. To investigate 19-nor-DOC's regulation and possible pathogenesis in hypertension, urinary free (UF) 19-nor-DOC was measured in 14 hypertensives, correlated with other corticosteroids and systemic arterial blood pressure (BP), and compared to basal and ACTH-stimulated values in 8 normotensive subjects. Seven of the 14 hypertensives had low-renin hypertension, 2 had primary aldosteronism, 1 had an adrenal carcinoma, and another had acromegaly. These studies determined that: 1) although the mean UF 19-nor-DOC was not increased in hypertensives (588 +/- 180 vs. 428 +/- 122 ng/day), 2 low-renin hypertensives had quite elevated levels (2186 and 2018); 2) the UF 19-nor-DOC in hypertensives was correlated with BP but not with PRA, aldosterone secretion, plasma potassium, basal plasma cortisol, or 17-hydroxycorticosteroids; 3) likewise, in normotensives, UF 19-nor-DOC did not correlate with basal plasma cortisol, cortisol secretion, or 17 hydroxycorticosteroids excretion but did correlate after ACTH stimulation. Therefore, although 19-nor-DOC is activated by ACTH administration, it is not correlated with basal parameters of cortisol production, suggesting that factors other than ACTH regulate basal 19-nor-DOC secretion. Furthermore 19-nor-DOC is elevated in some hypertensive patients, and it is directly related to the elevation of mean systemic BP. This suggests that, although 19-nor-DOC could contribute to hypertensive disease in some individuals, it does not appear to be due to excess ACTH. PMID- 6292260 TI - Protection against experimental allergic neuritis with silica quartz dust. AB - Lewis rats sensitized against peripheral nervous system antigens can be protected against experimental allergic neuritis (EAN) by the intraperitoneal injection of silica quartz dust. Two doses of 200 mg silica given 8 and 11 days post inoculation (dpi) protected against the development of both clinical and pathological disease. A single dose of 200 mg silica 8 dpi gave significant protection against clinical disease but all animals developed pathological signs. A single injection of 200 mg silica 11 dpi, after the onset of early signs, protected against further progression of disease. The protection was long lasting. Given the known toxic effects of silica for macrophages, these results would support the conclusion that macrophages function during the effector stage in the clinical and pathological expression of EAN in the Lewis rat. PMID- 6292261 TI - A prospective study of cytomegalovirus and herpes simplex virus disease in renal transplant recipients. AB - A prospective study of 84 renal graft recipients demonstrated cytomegalovirus (CMV) disease after transplantation in 37% of patients. Reactivation infection was found in 20 of 44 patients (46%) who were seropositive for CMV prior to transplant and primary CMV disease occurred in 11 of 40 (28%) initially seronegative patients. Nearly all cases of primary disease (91%) were associated with symptoms and in these cases CMV was probably acquired via the donated kidneys. Only 35% of the reactivation infections were associated with clinical symptoms. Actuarial life tables indicated that CMV disease did not reduce the length of graft survival. Herpes simplex virus (HSV) infections were diagnosed in 44 (52%) of the patients and included a fatal case of disseminated disease associated with hepatitis. PMID- 6292262 TI - Tick-borne fever and concurrent parainfluenza-3 virus infection in sheep. PMID- 6292263 TI - A hepatocellular adenoma in a rattlesnake. PMID- 6292259 TI - The biology of the human natural killer cell. AB - Natural killer (NK) cells in the human are a population of large granular lymphocytes (LGL) with at least one unique surface antigen not expressed on cells of other lineages. NK-target-cell interaction appears to involve carbohydrate recognition and, following binding, the NK cells are induced to generate O2-, transmethylate membrane phospholipids, and activate phospholipase A2. Some or all of these activities trigger a cascade of events which ultimately leads to the secretion of a substance toxic to the target cell. A variety of genes controls various steps in this cytolytic pathway. There is a good deal of evidence in the mouse, and some in the human, that NK cells play a role in host surveillance against tumor development, resistance to viral infections, and, possibly, hematopoietic regulation. PMID- 6292264 TI - Aspects of heredity, syndromic associations, and course of conditions in which cutaneous lesions occur solitarily or in multiplicity. AB - Many tumors of the skin, whether originating from ectoderm or mesoderm, exist in either single or multiple form. In the multiple state there are frequently associations with other cutaneous and systemic abnormalities, sometimes serious and even life-threatening. The resultant syndrome is almost always inherited in an autosomal dominant pattern. By contrast, solitary lesions are not heritable, nor are they associated with other stigmata. The solitary forms appear in middle or late life, the multiple, syndromic forms in childhood or early adulthood. PMID- 6292265 TI - Cutaneous lesions heralding disseminated cytomegalovirus infection. PMID- 6292266 TI - Behavioral detection of subcortical stimuli: comparison of somatosensory and "motor" circuits. AB - Cats were trained to press a lever for food reinforcement in response to stimulation of the ventral lateral (VL) nucleus of the thalamus and the deep cerebellar nuclei. By scaling stimulus intensities relative to the appearance of a minimal amplitude evoked response in pericruciate cortex, it was possible to measure behavioral detection thresholds and correlate behavior with electrocortical activity. With stimulus rates of 25 Hz or greater, VL was the least effective stimulus site for producing detection. At stimulus rates less than 25 Hz, stimulation of the lateral or interpositus nuclei was even less effective in eliciting behavior, but at rates of 25 Hz or more, detection thresholds decreased below those for VL stimulation; cerebellar stimulation produced detection as readily as had stimulation of the ventrobasal complex in other experiments. These findings suggest that the cerebellum may modulate sensory experiences and that some portions of cerebral cortex, the pericruciate and suprasylvian regions, do not appear to be directly involved in mediating sensory detection. It is postulated that the neural detection circuits are more likely to be found in subcortical than in cerebrocortical structures. PMID- 6292267 TI - The hamster hippocampal slice: I. Physiological properties. AB - The physiological properties of the in vitro hippocampal slice of the golden hamster are described. Hamster hippocampal tissue displays features similar to those seen in other species in terms of postsynaptic response characteristics to stimulation of monosynaptic afferents in the three primary subfields of the hippocampus. This pattern of physiological similarity supports the contention of the uniformity of hippocampal function across species, an important consideration with regard to the role of the hippocampus in brain and behavioral function. These results will permit the utilization of the hamster hippocampal slice in neurophysiological and neuroendocrine studies. PMID- 6292268 TI - Potentiation of odor by taste in rats: tests of some nonassociative factors. AB - The contribution of nonassociative neophobia and sensitization to the potentiation of odor by taste in rats was tested in three experiments. In Experiment 1, neophobia for almond odor (O), saccharin taste (T), and odor-taste compound (OT) cues was tested before and after noncontingent lithium chloride poisoning and compared with conditioned aversions produced by OT-LiCl temporal pairing. The OT compound potentiated unconditioned neophobia, but there was no evidence of poison-enhanced neophobia, disinhibition of neophobia, or sensitization by noncontingent LiCl; temporal pairing produced aversions for the compound and its elements. In Experiment 2, generalization to a novel odor was tested after O-LiCl or compound OT-LiCl pairing. The potentiated odor aversion did not generalize to the novel odor; it was specific to the odor paired with taste and LiCl. In Experiment 3, potentiation of the odor component by a discriminant or nondiscriminant taste component was tested. Potentiation was evident only when a novel discriminant taste was in compound with odor prior to LiCl poisoning. These studies support an associative "indexing" hypothesis of the potentiation effect in rats. PMID- 6292269 TI - Pavlovian conditioning with ethanol and lithium: effects on heart rate and taste aversion in rats. AB - Rats received paired injections of either ethanol or saline as the conditioned stimulus and lithium chloride as the unconditioned stimulus (US) in a Pavlovian differential conditioning paradigm. Lithium chloride evoked a large deceleration in heart rate (80-100 beats per minute) as an unconditioned response. As a result of 10 conditioning trials, the substance paired with LiCl elicited a lower average heart rate than that elicited by the unpaired substance. Moreover, animals that received ethanol-LiCl injections subsequently were more averse to the taste of ethanol than animals receiving saline-LiCl pairings. However, there were no differences in ethanol's ability to serve as the US to induce an aversion to a novel flavor solution (i.e., the Avfail phenomenon was not observed). The overall pattern of results underscores the value of using multiple indexes of learning in drug-drug conditioning paradigms. PMID- 6292270 TI - Acquisition and extended retention of a conditioned taste aversion in preweanling rats. AB - The time course of memory decay for infant rats may shed light on the processes responsible for infantile amnesia. A taste aversion conditioning procedure appropriate for both neonatal and adult rats was employed in four experiments to investigate the ontogeny of extended retention. In Experiment 1, rats trained at 1, 10, 20, or 60 days of age were tested for retention of the taste aversion 25 days later. At testing, only those rats conditioned when 20 or 60 days old demonstrated significant taste aversions. Experiments 2 and 3 established that rats 14-15 days old and older were able to retain significant taste aversions following a 25-day retention interval. Younger rats did, however, acquire and retain the aversion for several days and showed a gradual retention loss over progressively longer retention intervals (Experiment 4). These findings suggest that preweanling rats demonstrate initial consolidation, storage, and retrieval of conditioned taste aversions. It is only after this initial period that retention deficits become evident. PMID- 6292271 TI - Malignant fibrous histiocytoma of the spermatic cord. PMID- 6292272 TI - Computed tomography of the brain, liver, and upper abdomen in the staging of small cell carcinoma of the lung. AB - Computed tomography (CT) examinations of the brain, liver, and upper abdomen were obtained in 60 previously untreated patients with histologically proven small cell carcinoma of the lung (SCCL). These results, together with clinical findings and laboratory investigations including radionuclide scans, determined the final staging of the patients. Computed tomography changed the stage from limited to extensive disease in three patients and from extensive to limited disease in seven patients. Overall, 10 of 60 patients (16%) had their final stage altered by CT. This study indicates that CT examinations may yield valuable information in some cases of small cell lung cancer but that it is of limited value in the routine staging of these patients. PMID- 6292273 TI - Epidermoid hidradenoma. A clinicopathologic study. AB - Clinical and histopathologic data in 8 cases of epidermoid hidradenoma are presented. Most of the tumors were in the head and neck, and most of the patients were middle-aged and older adults. Generally, the lesions were asymptomatic nodules that sometimes showed ulceration or rapid growth. Despite having some histologic atypia or focal mitotic activity, the tumors were found to be benign on long-term evaluation. This epidermoid variant should not be otherwise differentiated from the benign group of solid-cystic hydradenomas, because cytologic variability did not predict a significant change in prognosis. PMID- 6292274 TI - Extramammary Paget's disease in fibroepithelioma of Pinkus. PMID- 6292275 TI - Concentration of foot-and-mouth disease virus in milk of cows infected under simulated field conditions. PMID- 6292276 TI - Ion transport through the enamel organ--an update. PMID- 6292277 TI - Mechanisms by which the enamel organ controls calcium entry into developing enamel. PMID- 6292278 TI - [Common mechanism of the intratissue regulation of proliferation on the principle of the tissue-specific control of mitochondrial oxidative phosphorylation]. PMID- 6292279 TI - Selective subsensitization of beta-adrenergic receptors in central airways of asthmatics and normal subjects during long-term therapy with inhaled salbutamol. AB - In five subjects with mild asthma and in five normal subjects, we determined the effect of a 4 wk course of inhaled salbutamol (albuterol), 200 micrograms q.i.d., on (1) acute bronchodilator responsiveness, (2) bronchial sensitivity to inhaled histamine, (3) beta-adrenergic protection against histamine-induced bronchospasm, and (4) beta-receptor density of peripheral blood lymphocytes. We observed a diminution in central airway bronchodilator responsiveness (as measured by airway conductance responses) to acutely inhaled salbutamol and to subcutaneous terbutaline in both groups of subjects, although only the response to subcutaneous terbutaline was statistically significant (p less than 0.02). On the other hand, no impairment of small airway bronchodilator responsiveness was noted in either group of subjects when responses were measured as partial expiratory flow rates at 60% below total lung capacity. These findings suggest the development of selective subsensitization of beta-receptors in the larger central airways, where a proportionately greater amount of the inhaled beta-agonist aerosol would necessarily be deposited. A greater loss of protection against histamine-induced bronchospasm was seen in asthmatics than in normals (approximately twofold), although the difference was not significant. A modest but not significant reduction in peripheral blood lymphocyte beta-receptor density was observed by the end of the 4 wk treatment period. The possibility that the observed changes in bronchodilator responsiveness might influence the morbidity and mortality associated with bronchial asthma is discussed. PMID- 6292280 TI - Effect of sustained-release theophylline administration on pituitary-thyroid axis. AB - The effect of therapeutic doses of theophylline on the pituitary-thyroid axis was studied in four normal adults and in 15 asthmatic children on theophylline treatment. In the four normal adults, sustained-release theophylline was administered every 12 hr for 60 hr, and serial determinations of serum theophylline, thyroxine (T4), triiodothyronine (T3), reverse triiodothyronine (rT3), thyrotropin (TSH), and cyclic AMP (cAMP) were done. In all four cases, serum T4 increased significantly (134% +/- 4% vs basal level) with significant correlation to plasma cAMP level (r = 0.67, p less than 0.05). Changes in T3 and rT3 showed a marked individuality; an increase in T3 occurred in a subject with severe side effects, while the increase in rT3 was observed in a subject with mild side effects. Of the 15 children on theophylline treatment, a higher T4 level was found in 12 and a lower T3 level in nine at 1 wk, but these values returned to the previous level after 4 wk. These results indicate that the therapeutic doses of theophylline significantly but transiently increased the serum T4 level. In most cases, T4 is metabolized to rT3, an inactive metabolite of T4, but in a few cases it is metabolized to T3, which might double the side effects of theophylline. PMID- 6292281 TI - [Postovulatory hormones in plasma, peritoneal and follicular compartments]. AB - Eleven women with dysovulatory or unexplained sterility were treated by induction of ovulation and compared with a control group of seven normal women. Laparoscopy was carried out 48 hours after HCG had been injected or after the LH peak in the first group. 48 hours after the rise in temperature in the second group ovulation was presumed and aestradiol 17 beta, progesterone. Delta 4 androstenedione, FSH, LH, prolactin and cyclic AMP estimated in the plasma, the peritoneal fluid and fluid from the persistent follicles. A corpus luteum of recent formation was found in group 1, 5 out of 7 times, and only 7 out of 11 times in group 2. The question of whether non-ruptured luteinised follicles exist is discussed in view of these anatomical and hormonal level findings. These show that there is a good correlation in both groups between aestradiol 17 beta and progesterone, whether estimated in the plasma or in peritoneal fluid. On the other hand, there was poor evidence of correlation between the levels of steroids in each compartment as compared with the other. Finally, the curves for the different steroid levels are important when concentrations of hormones in the follicles, the peritoneal fluid and the plasma are compared. Conclusions have been suggested about the origin of the different steroids and their movement from compartment to compartment. PMID- 6292283 TI - Laboratory studies on monkeypox virus. PMID- 6292282 TI - The development of beta-adrenergic mediated inhibition of growth hormone secretion in the ovine fetus. AB - The ontogeny of the suppressive effect of the beta-adrenergic agonist, isoprenaline, on fetal growth hormone (GH) release was examined in 14 chronically catheterized ovine fetuses. Isoprenaline was administered as an intravenous infusion over 1 h (200 micrograms/kg). In seven fetuses between 72 and 99 days of gestation, isoprenaline had no effect on fetal plasma GH concentrations. In seven older fetuses between 114 and 140 days of gestation, isoprenaline infusion suppressed (P less than 0.02) fetal GH release. No effect was observed in five saline-treated control fetuses (119-131 days). Propranolol (250 micrograms/kg i.v.) administered 5 min prior to the isoprenaline infusion to four fetuses (117 136 days) delayed (P less than 0.05) the onset of the suppressive effect of isoprenaline demonstrating that the action of isoprenaline was mediated by the beta-adrenergic receptor. Propranolol alone (n = 6) had no effect. These observations demonstrate that the potential for beta-adrenergic inhibition of fetal GH release differentiates after 100 days of gestation. Comparison with previous studies of the ontogenesis of the control of GH secretion suggests that the hypothalamic beta-adrenergic control of GH release differentiates with an intermediate time course compared to other potential neuroendocrine controls. PMID- 6292284 TI - Pesticide-induced changes in hepatic microsomal enzymes in CF-1 mice. PMID- 6292285 TI - Immunohistochemical localization of tonin and its relation to kallikrein in rat salivary glands. AB - Tonin and kallikrein are serine proteases present in high concentrations in the submandibular gland of the rat. These enzymes release the vasoactive peptides angiotensin II and lysyl-bradykinin from the precursors angiotensinogen and kininogen, respectively. Tonin and kallikrein were purified from homogenates of rat submandibular gland, and antisera against each protein were raised in rabbits. The anti-kallikrein antibody also reacted with tonin, showing partial cross-reactivity between kallikrein and tonin when tested by double immunodiffusion and by immunoelectrophoresis. The anti-tonin antibody did not appear to react with kallikrein in immunodiffusion systems. The cellular localization of tonin was investigated by the indirect immunofluorescence and the peroxidase-antiperoxidase techniques. In the granular tubular cells tonin specific staining was abundantly present with a granular distribution; in the striated duct cells tonin-specific staining was observed as a thin luminal rim. Tonin was not detected in any other structures of the gland. When the localization of tonin was compared with that of kallikrein, both enzymes were found within the same granular tubular cells. However, more kallikrein than tonin was detected in the striated duct cells. Furthermore, kallikrein but not tonin was found in the ductal cells of the parotid and sublingual glands. PMID- 6292286 TI - Adenylate cyclase localization in unfixed specimens of rat oral mucosa and isolated mitochondria. AB - The cytochemical localization of adenylate cyclase (E.C.4.6.1.1) was demonstrated in unfixed rat oral mucosal specimens and isolated mitochondrial fractions by use of a modified cytochemical reaction mixture. The adenylate cyclase reaction product was observed in the mitochondrial cristae of intact epithelial and fibroblast cells, and in isolated mitochondria after a 10-min incubation in the reaction mixture. The validity of the cytochemical adenylate cyclase method was supported by biochemical studies. PMID- 6292287 TI - Immunologic studies of aging. VIII: no change in cyclic nucleotide concentration in T lymphocytes from old humans despite their depressed proliferative response. PMID- 6292288 TI - Lyt phenotype of H-2b CTL effectors and precursors specific for the SV40 transplantation rejection antigen. AB - The immune effector cells mediating the in vitro immune response to the SV40 transplantation rejection antigen were characterized by using monoclonal antibody directed against lymphocyte differentiation (Lyt) antigens. Two distinct T lymphocyte populations were found to be responsible for the in vitro lysis of SV40-transformed cells, and Lyt 1+,2+ cytotoxic T lymphocyte (CTL) that is present in the spleens of SV40 immunized mice 9 days postimmunization and an Lyt 1-,2+ CTL that is generated by secondary in vitro stimulation of in vivo primed spleen cells. During secondary in vitro stimulation of SV40 immune spleen cells obtained 9 days postimmunization, a shift in the Lyt phenotype of the CTL from Lyt 1+,2+ to Lyt 1-,2+ is observed. Although the Lyt 1-,2+ CTL can be derived from Lyt 1-,2+ noncytotoxic memory cells, it is not known whether the Lyt 1+,2+ CTL differentiates into an Lyt 1-,2+ CTL during in vitro stimulation. PMID- 6292289 TI - An adherent cell lyses virus-infected targets: characterization, activation, and fine specificity of the cytotoxic cell. AB - We have previously reported that mice with defective T lymphocyte function clear a primary reovirus type 1 infection in a normal fashion. The present studies were initiated to determine what cells may play a role in the clearance of this primary infection. We show that macrophages from unprimed mice are capable of lysing reovirus type 1-infected target cells in the absence of specific antibody. Macrophages from nu/nu mice have higher levels of this lytic activity than macrophages from nu/+ and normal mice. In addition, PEC from endotoxin nonresponsive C3H/HeJ mice have virtually no anti-viral lytic activity, while PEC from C3H/FeJ mice have high levels of such activity. Incubation of PEC from C3H/HeJ mice overnight in Con A supernatants restores this lytic activity. PEC are capable of lysing reovirus type 1-infected target cells but not those infected with type 3 reovirus. Using intertypic recombinant viruses, we show this striking target cell specificity to be a property of the sigma 1 protein, the viral hemagglutinin. PMID- 6292290 TI - Immune deficiency in the X-linked lymphoproliferative syndrome. I. Epstein-Barr virus-specific defects. AB - Eleven males with XLP were evaluated for EBV-specific antibodies during periods of 2 to 7 yr. Variable responses to EBV-specific antigens were found. All 11 patients had subnormal anti-EBNA titers, which probably reflected a T cell deficiency. The patients showed four different patterns in their anti-VCA response: 1) two boys who had experienced malignant lymphoma mounted no antibodies at all; 2) two patients showed intermittent anti-VCA titers; 3) four males had persistently elevated anti-VCA titers; and 4) three patients showed normal anti-VCA titers. ADCC against EBV-infected cells was abnormally low in six patients and was elevated in two patients given gamma-globulin. ADCC titers did not correlate with anti-VCA titers. However, most patients with XLP failed to effect regression of autologous EBV-infected lymphoblastoid cell lines, indicating a deficiency in long-lived T cell-mediated immunity to EBV. PMID- 6292291 TI - Immune deficiency in the X-linked lymphoproliferative syndrome. II. Immunoregulatory T cell defects. AB - Surface phenotypic markers and the function of lymphocytes in patients affected with the X-linked lymphoproliferative syndrome (XLP) were studied. This syndrome is characterized by a defective response to infection with Epstein Barr virus (EBV). Normal numbers of B and T cells were detected with anti-Ig and monoclonal OKT3 antisera, respectively. T cell subset values, however, were persistently altered: cells reacting with OKT8 were significantly elevated in five of nine patients, accompanied by a slight decrease in the percentage of OKT4-positive cells, leading to abnormally low OKT4 to OKT8 ratios. One patient had a high OKT4 to OKT8 ratio due to low number of OKT8-positive cells. Lymphocytes from patients showed normal proliferation after stimulation with T and B cell mitogens. In contrast, Ig synthesis by lymphocytes after stimulation with B cell mitogens was markedly deficient: low or undetectable levels of one or all classes of Ig were detected, whereas cell lines established from EBV-infected B lymphocytes from patients produced normal quantities of Ig. These studies imply immune regulatory impairments in the patient with XLP. PMID- 6292292 TI - Glycosylation and secretion of an altered immunoglobulin heavy chain in mouse myeloma MOPC 315. AB - A variant line (LV-1) of mouse myeloma MOPC 315 (IgA, lambda 2) has lost the ability to synthesize L chain. It synthesizes an altered H chain (H' chain) that is turned over intracellularly and is not secreted. Rescue of H' chain secretion can be accomplished by fusion of LV-1 to a variant of another myeloma line, MPC 11 (IgG2b, kappa), which only synthesizes a light chain. The hybrid (X-2) secretes the H' chain in a four chain structure (kappa 2 alpha' 2). In wild-type MOPC 315 cells, it was reported previously that inhibition of core sugar addition blocks the secretion of the H chain polypeptide. We have studied glycosylation in MPOC 315 wild-type, LV-1 variant, and X-2 hybrid cell lines. The ability of all three lines to add the core sugars mannose and glucosamine to heavy chain was demonstrated. Due to the instability of the H' chain in LV-1, it is difficult to assess H' chain fucosylation directly. To study fucose addition in LV-1, the enveloped virus vesicular stomatitis (VSV), which can infect the three lines, was utilized. The fucosylation and secretion of VSV glycoprotein G was discernible in all three lines; however, only LV-1 cannot activate free fucose, and instead fucosylates through conversion of the mannose intermediate. Normal fucose addition to H chain in a wild-type cell occurred immediately before secretion. The fact that degradation of H' chain in LV-1 begins before fucosylation suggests that the rescue of H' chain secretion by formation of the X-2 hybrid is due to the acquired presence of a suitable L chain rather than complementation of a sugar defect. These observations indicate that proper assembly of the polypeptide components of some secretory proteins, e.g., Ig molecules, is required for the secretion of the individual chains. PMID- 6292293 TI - Cyclic GMP as the second messenger in helper cell requirement for gamma interferon production. AB - Cyclic GMP and activators (acetylcholine, E. coli heat-stable toxin) of guanylate cyclase were capable of completely replacing the helper cell or interleukin 2 requirement for gamma-interferon (IFN gamma) production by Lyt-1-,2+ cells from C57BL/6 mouse spleen cells. The cyclic GMP help was independent of DNA synthesis or proliferation in the IFN gamma-producing cells, because cyclic GMP reversed mitomycin C blockage of IFN gamma production but did not reverse the inhibition of DNA synthesis. Thus, the findings presented here are unrelated to the question of the second messenger role of cyclic GMP in the activation of lymphocytes for DNA synthesis and cellular proliferation. The cyclic GMP help for IFN gamma production was antagonized by cyclic AMP and inducers (isoproterenol) of adenylate cyclase. PMID- 6292294 TI - Structure of the murine plasma cell alloantigen PC-1: comparison with the receptor for transferrin. AB - The plasma cell alloantigen PC-1 was isolated from C1.18 myeloma cells by immunoprecipitation and was analyzed by polyacrylamide gel electrophoresis. It was found to consist of two similar or identical disulfide-bonded polypeptide chains, each of Mr 115,000. The mobility of PC-1 in nonequilibrium pH gradient electrophoresis was similar to that of bovine serum albumin (pI 4.9). The PC-1 antigen is therefore similar to the transferrin receptor in Mr, charge, subunit composition, disulfide bonding, and developmental regulation. Similarities can also be detected by peptide mapping with subtilisin, but not with staphylococcal V8 protease. It is suggested that the PC-1 protein and the transferrin receptor may have had a common evolutionary origin, and may have similar functions. PMID- 6292295 TI - T lymphocyte responses of murine lung: immunization with alloantigen induces accumulation of cytotoxic and other T lymphocytes in the lung. PMID- 6292296 TI - Augmentation of phagocytosis by a specific fibronectin fragment that links particulate activators to the fibronectin adherence receptor of human monocytes. AB - Intact human plasma fibronectin of 44,000 m.w. and a fibronectin fragment of 180,000 m.w. promote dose-dependent adherence of gelatin-coated particles to human monocytes without phagocytosis. Both of these proteins, however, augment monocyte ingestion of gelatin-coated targets that are particulate activators of the alternative complement pathway or of nonactivators bearing IgG. Unlike intact fibronectin, the 180,000 m.w. fragment also binds directly to particulate activators that lack gelatin to augment their phagocytosis by human monocytes. Prior attachment to monocytes of gelatin-coated sheep erythrocytes bearing increasing concentrations of intact fibronectin decreases in a dose-dependent fashion the capacity of these monocytes to engage in augmented phagocytosis of particulate activators opsonized with the 180,000 m.w. fibronectin. Occupation of the monocyte fibronectin receptors with particle-bound, intact fibronectin does not decrease monocyte phagocytosis of plain particulate activators or of IgG coated particles. Thus, the 180,000 m.w. fibronectin fragment both directly opsonizes particulate activators and interacts with monocyte fibronectin receptors to promote particle adherence, thereby enhancing phagocytosis through a concerted action with the distinct receptors for particulate activators. PMID- 6292297 TI - Biochemical signals transmitted by Fc gamma receptors: triggering mechanisms of the increased synthesis of adenosine-3',5'-cyclic monophosphate mediated by Fc gamma 2a- and Fc gamma 2b- -receptors of a murine macrophage-like cell line (P388D1). AB - The specific binding of IgG2a or IgG2b subclass monoclonal anti-sheep erythrocyte antibodies to P388D1 cell surface Fc gamma 2aR3 or Fc gamma 2bR, respectively, triggered the synthesis of adenosine-3'5'-monophosphate (cAMP) to an approximately same extent by the mechanisms that are apparently unique for each type of Fc gamma Rs. Fc gamma 2aR appeared to trigger directly, upon binding of IgG2a antibodies, the adenylate cyclase system without requiring the participation of guanine nucleotide-binding (G/F) regulatory protein, because the Fc gamma 2aR-triggered cAMP synthesis, which reached maximum within 30 min, was not significantly affected by an uncoupler, Mn++ or by addition of guanosine triphosphate (GTP) analog, 5'-guanylylimidodiphosphate (Gpp(NH)p). In contrast, Fc gamma 2bR appeared to stimulate indirectly the G/F regulatory requiring adenylate cyclase system by generating prostaglandins, since the cAMP synthesis, which required 90 min to reach plateau after binding of IgG2b to Fc gamma 2bR, was totally suppressed by phospholipase A2 inhibitor (p-bromophenacylbromide) or cyclo-oxygenase inhibitor (indomethacin), partially suppressed by Mn++, and slightly increased by Gpp(NH)p. Furthermore, the inhibition of phagocytic process by cytochalasin D increased cAMP synthesis mediated by Fc gamma 2aR (about 70% at 2 micrograms/ml), but did not affect Fc gamma 2bR-mediated cAMP synthesis. In addition, our data suggested that both Fc gamma 2aR- and Fc gamma 2bR-mediated cAMP synthesis are independent from beta-adrenergic receptor-mediated stimulation of the adenylate cyclase system, since either beta-agonist (isoproterenol) or beta-antagonist (propranolol) did not affect significantly the levels of cAMP produced in response to EA-stimulation. PMID- 6292298 TI - Bromination of guanosine and cyclic GMP confers resistance to metabolic processing by B cells. AB - The metabolic fates of 8-bromoguanosine (8BrGuo) and 8-bromoguanosine-3'5'-cyclic monophosphate (8Br-cGMP) were examined in cultures of murine B lymphocytes. These compounds exert striking immunostimulatory effects upon bone marrow-derived lymphoid cells in vitro. Both 8BrGuo and 8Br-cGMP were resistant to metabolic processing by these cells. That purine metabolic pathways are intact and operant in B cells was demonstrated by the ready degradation and phosphorylation of native guanosine and cyclic GMP. Inaccessibilty of the substrate to the relevant enzymes was ruled out as an explanation by the observation that the brominated compounds also were resistant to processing in broken cell preparations. Moreover, 8BrGuo did not interfere with the cellular machinery for metabolizing native guanosine. The implications of these observations for studying the actions of purine nucleotides, cyclic nucleotides, and their enzymatic processing in B cells are discussed. PMID- 6292299 TI - Inhibitory action of elevated levels of adenosine-3':5' cyclic monophosphate on phagocytosis: effects on macrophage-Trypanosoma cruzi interaction. AB - The effects of agents that elevate intracellular levels of cyclic AMP on the in vitro interaction of mouse peritoneal macrophages with virulent bloodstream forms of a reticulotropic strain of Trypanosoma cruzi were investigated as a part of our efforts to define the requirements for tissue invasion by this intracellular human pathogen. At optimal, non-toxic concentrations, both L-isoproterenol and prostaglandin E1, agents that increase cyclic AMP levels by activating adenylate cyclase via different mechanisms, reduced the uptake by T. cruzi by the macrophages by approximately 30 and 70%, respectively, and also caused a reduction in the numbers of macrophages capable of incorporating the parasites. Similar results were obtained when either theophylline, which increases cyclic AMP levels by inhibiting the catabolic effect of phosphodiesterase activity, was incorporated into the cultures or by direct addition of dibutyryl cyclic AMP. Reductions produced with optimal concentrations of these drugs amounted to approximately 40 to 50%. The present results indicate that binding and uptake of virulent forms of a reticulotropic strain of T. cruzi by macrophages from a susceptible host are under the regulatory influence of cellular cyclic AMP levels. PMID- 6292300 TI - Two different rheumatoid factor-producing cell populations distinguished by the mouse erythrocyte receptor and responsiveness to polyclonal B cell activators. AB - The subsets of human peripheral blood B lymphocytes from which Epstein Barr virus (EBV) and pokeweed mitogen- (PWM) induced IgM anti-IgG autoantibody-producing B cells arise have been compared. EBV-induced IgM anti-IgG autoantibodies preferentially derive from a subset of B cells that forms rosettes with mouse erythrocytes. In contrast, PWM-induced IgM anti-IgG antibodies preferentially arise from a B cell subset lacking the mouse erythrocyte receptor. PMID- 6292301 TI - Human T cell hybridomas specific for Epstein Barr virus-infected B lymphocytes. AB - A mutant of the Jurkat human T lymphoblastoid cell line deficient in hypoxanthine phosphoribosyltransferase, and resistant to ouabain, was fused with peripheral blood T lymphocytes primed in vitro with Epstein Barr virus- (EBV) transformed autologous B lymphocytes. After selection of somatic cell hybrids and cloning, hybridoma cell lines were obtained that reacted with autologous EBV-infected B lymphocytes, as detected by the release of interleukin 2 into the culture medium. The hybridomas did not react with i) EBV-uninfected autologous or allogeneic B lymphocytes, ii) three out of four allogeneic EBV-transformed cell lines, or iii) two established EBV-negative B cell lines. These functional hybridomas may ultimately prove useful in dissecting the means by which human T lymphocytes recognize and regulate EBV infection in vivo. PMID- 6292302 TI - Cytotoxicity by NK-like cells from hepatitis B-immune patients to a human hepatoma cell line secreting HBsAg. PMID- 6292303 TI - Induction of human IgE synthesis by a factor derived from T cells of patients with hyper-IgE states. AB - The requirements for the induction of IgE synthesis in normal B cells were studied. In contrast to peripheral blood lymphocytes (PBL) from allergic subjects, normal PBL failed to synthesize IgE spontaneously in vitro. Pokeweed mitogen (PWM) and Epstein Barr virus (EBV) failed to induce IgE synthesis in normal PBL and in tonsil lymphocytes. In the case of PWM, this failure was not overcome by prior removal of T8+ cells, which were shown previously to contain IgE-specific suppressor cells. In the case of EBV, the failure to induce IgE synthesis was not overcome by prior removal of sheep rosette-forming cells. Supernatants of T cells derived from three groups of patients with elevated serum IgE (hyper-IgE syndrome, atopic dermatitis, and acute graft-vs-host disease) induced significant IgE synthesis in cultures of normal B cells without causing an increase in IgE synthesis. In contrast, supernatants of normal T cells failed to induce IgE synthesis. Release of the IgE isotype-specific helper factor was inhibited by cycloheximide and tunicamycin, and its activity was destroyed by treatment with trypsin and neuraminidase. These results indicate that whereas classical polyclonal B cell activators (PWM, EBV) fail to induce IgE synthesis by normal B cells, IgE synthesis is readily induced by an IgE-specific helper factor released by T cells from patients with hyper-IgE states. PMID- 6292304 TI - Functional properties of T lymphocytes and their subsets in cytomegalovirus mononucleosis. AB - Cytomegalovirus (CMV) mononucleosis is associated with diminished lymphocyte responses to mitogens and herpes virus antigens, and a reversal in the normal ratio of helper-inducer (OKT4-positive) to cytotoxic-suppressor (OKT8-positive) cells. In this report, we investigate the proliferative and cytotoxic functions of lymphocyte subpopulations from patients with CMV mononucleosis. Both mitogen responsiveness and the ability to generate cytotoxic lymphocytes against allogeneic Laz 156 cells in vitro were markedly diminished. Mononuclear cells enriched for OKT8+ lymphocytes showed little or no responsiveness to Con A; cells enriched for OKT4+ lymphocytes were considerably more responsive. During CMV mononucleosis, the predominant T cell subset is a hyporesponsive OKT8+ cell. In vitro culture of peripheral blood lymphocytes, however, resulted in a selective loss of OKT8+, OKIa+ cells, so that the remaining T cell population was composed primarily of Con A-responsive OKT4+ cells. Convalescence was associated with the return of OKT8+ responsiveness to Con A, a greater stability of OKT8+ cells during culture, and the partial recovery of ability to develop cytotoxic lymphocytes after in vitro sensitization. PMID- 6292305 TI - Malignant rabbit fibroma virus causes secondary immunosuppression in rabbits. AB - Shope fibroma virus (SFV) causes a localized, self-limited, fibroblastic proliferation in adult rabbits. Extracts of Shope fibroma tumors were found to contain a second virus that induces a rapidly progressive disseminated tumor. Dissemination of this malignant fibroma is associated with activation of commensal mucosal infection with Pasteurella multocida, causing purulent conjunctivitis and rhinitis and resulting in death from nasal obstruction. We have isolated this new agent by two cycles of plaque purification. It is a poxvirus that is antigenically virtually identical to SFV as measured by a plaque reduction assay, but behaves differently both in vivo and in vitro. We have called this virus malignant rabbit fibroma virus (MV). Electrophoresis of restriction digests made with HIND III indicates that despite the antigenic similarity of SFV and MV, the locations of HIND III sites in the two viral genomes are quite different. These experiments have enabled us to determine that MV was present in small quantities in our initial uncloned stock of Patuxent strain SFV. Lymphocytes from rabbits bearing MV-induced tumors responded poorly to both B and T lymphocyte mitogens. This nonspecific immunologic dysfunction is evident at or before the time when metastases and Gram-negative infection develop, and it becomes more profound as the disease progresses. MV-induced tumors may provide a model for Gram-negative infection and decreased immunologic responsiveness associated with malignancies. PMID- 6292306 TI - An investigation of the use of urease-antibody conjugates in enzyme immunoassays. AB - The development of urease (E.C.3.5.1.5) as a label for enzyme immunoassay (EIA) procedures is described and the use of such conjugates illustrated with examples. Urease catalyzes the hydrolysis of urea to carbon dioxide and ammonia. The production of ammonia may be detected readily by a pH shift which we have found best indicated by the vivid colour change (yellow to purple) of bromocresol purple incorporated in the substrate solution. This enzyme-substrate system offers a number of important advantages. The substrate in aqueous solution is stable, titration end points are sharp and readily visible and the enzyme is not inhibited by sodium azide. Thus, test reagents may be prepared with this preservative and stored ready to use. Urease of high specific activity is commercially available and because it does not occur in mammalian tissues, it is suitable for use in EIA tests to detect cell-associated antigens and their antibodies. Finally, the enzyme reaction may be stopped by the addition of organomercurial preservatives, thus allowing storage of developed tests for later examination. PMID- 6292307 TI - A single-step centrifugation method for separation of granulocytes and mononuclear cells from blood using discontinuous density gradient of Percoll. AB - A rapid and reproducible method is described for the simultaneous purification of mononuclear and polymorphonuclear cells, based on the use of a discontinuous gradient of Percoll. Centrifugation at 600 X gav for 20 min resulted in separation of the mononuclear and polymorphonuclear cells into 2 distinct bands at the interfaces. The upper layer contained the mononuclear cells and the lower band highly purified polymorphonuclear cells. Both populations were purified about 97% without detectable cell alteration. PMID- 6292308 TI - Multiple adenomas of the liver. PMID- 6292309 TI - Colchicine-induced modulation of collagenase in human skin fibroblast cultures. I. Stimulation of enzyme synthesis in normal cells. AB - Microtubule-active agents affect the secretion of a variety of proteins, including collagenase. To gain insight into the mechanisms involved in this process, we examined the effects of colchicine on the synthesis, secretion, and activity of human skin collagenase. When added to monolayer cultures of human skin fibroblasts, 10(-6) M colchicine produced a mean 3-fold increase in trypsin activatable collagenase in the culture medium. Stimulation was not observed with lumicolchicine. The enhanced accumulation of collagenase was dose-dependent with 10(-9), 10(-8), 10(-7), and 10(-6) M colchicine giving collagenase activities/mg protein that were 100 +/- 6%, 165 +/- 20%, 186 +/- 34%, and 297 +/- 62% of control, respectively. Although the effect on collagenase was seen under conditions independent of cellular growth (i.e., in serum-free medium), maximum stimulation occurred in subconfluent cultures. The colchicine-induced increase in activity was paralleled by an increase in immunoreactive enzyme protein, suggesting stimulation of enzyme synthesis. The catalytic efficiency of the enzyme (activity per unit immunoreactive protein) was unchanged, however, indicating that a structurally normal enzyme was being synthesized. To examine the process in more detail, the biosynthesis of 3H-labeled collagenase was quantitated in these cultures by specific immunoprecipitation. Although 10(-6) M colchicine produced no increase in total protein synthesis, an increased rate of collagenase synthesis was seen after only 1.5 hr. These data suggest that colchicine has a specific effect on the synthesis of collagenase and may be a useful probe for studying its regulation. PMID- 6292310 TI - Colchicine-induced modulation of collagenase in human skin fibroblast cultures. II. A probe for defective regulation in epidermolysis bullosa. AB - The addition of colchicine to cultures of normal human skin fibroblasts produces a significant stimulation of collagenase. Because this finding implies a role for the microtubule system in the regulation of normal collagenase synthesis, we have used colchicine as a probe for aberrations in this enzyme in epidermolysis bullosa. In fibroblast cultures from the dominant simplex, dominant dystrophic, and recessive letalis forms of epidermolysis bullosa, 10(-6) M colchicine produced approximately a 2-fold increase in collagenase in the culture medium, a finding shown by biosynthetic studies to be attributable to enhanced synthesis of enzyme protein. In the case of typical recessive dystrophic epidermolysis bullosa, a disease characterized by excessive collagenase synthesis, the fibroblasts could also be stimulated to produce additional collagenase, despite having elevated baseline synthetic rates. In contrast, fibroblasts isolated from one recessive epidermolysis bullosa patient were resistant to the stimulatory effects of colchicine in concentrations up to 5 x 10(-6) M. In the absence of colchicine, collagenase synthesis in this patient's cells (termed REBc-) was 3-4 times that of normal controls, suggesting that the as yet undefined cellular function that is abrogated (or stimulated) by colchicine in normal cells may have been genetically impaired in these REBc- cells. Despite the resistance to colchicine, as manifested by the failure to stimulate collagenase, gross parameters of microtubular function, such as cell replication, were intact. Phenotypically, this patient had a form of epidermolysis bullosa intermediate between typical recessive dystrophic and recessive letalis forms of the disease. Although an experimentally induced blister was located in the lamina lucida, hypoplastic anchoring fibrils were also observed. These findings, in addition to the marked increase in collagenase synthesis, suggest the possibility that this patient may represent a compound heterozygote of two forms of epidermolysis bullosa and that colchicine may be useful in defining other such patients. PMID- 6292311 TI - Impairment of immunocompetent mouse spleen cell functions by infection with coxsackievirus B3. AB - Early after infection, mice inoculated with coxsackievirus B3 showed consistent reduction of antibody responsiveness. Beginning one week after infection they also evidenced progressive spleen atrophy. The cellular basis of the reduced antibody response exhibited in vitro by spleen cells of infected mice at the onset of atrophy was investigated by the use of different antigens, by supplementation with different subpopulations of immunocompetent cells from normal donors, and by cross-recombination with normal lymphoid cells. Whereas B cell functions appeared to be preserved, a deficit of macrophage accessory functions was clearly evident, possibly at the level of antigen presentation. In infected spleens, nonspecific suppressor T cells were also observed, but their activity did not correlate strictly with the degree of immunodepression. Because no evidence for a direct effect of virus on, or for viral replication in, immunocompetent cells was found, these alterations were tentatively ascribed to activation of the host's suppressor system. Such changes may have implications in the pathogenesis of coxsackievirus infections. PMID- 6292312 TI - Toxic shock syndrome. A review. PMID- 6292313 TI - Inactivation of lysosomal enzymes by the respiratory burst of polymorphonuclear leukocytes. Possible involvement of myeloperoxidase-H2O2-halide system. AB - Lysosomal enzyme release from PMN during phagocytosis was examined in PMN from healthy subjects and patients with CGD. Normal PMN showed increased extracellular enzyme levels and a marked reduction of total (intracellular plus extracellular) enzyme activity after phagocytosis of STZ. In PMN from patients with CGD, a reduction of total enzyme activity was not observed and much more enzyme than normal was released extracellularly. We have examined the effect of the respiratory burst on enzyme activity mostly to loss of released enzymes. PMA stimulated normal PMN inhibited the enzyme activity in postphagocytosis media from CGD PMN but PMA-stimulated CGD PMN did not. This inhibition by normal PMA stimulated PMN was protected by the addition of catalase, histidine, and NaN3. Furthermore, these enzymes were inactivated by cell-free, glucose plus glucose oxidase (as a H2O2 generating system) and NaOCI. Histidine had a protective effect on the enzyme inactivation both by glucose plus glucose oxidase and by NaOCI. In contrast, histidine gave a slight increase of extracellular enzyme levels but failed to restore enzyme activity when added to the medium during phagocytosis in normal PMN. These evidences suggest that the inactivation of lysosomal enzymes is mainly due to MPO-H2O2-halide system, resulting in inhibition of lysosomal enzyme release from PMN during phagocytosis. PMID- 6292315 TI - Multiple primary bronchogenic carcinomas. PMID- 6292314 TI - Stimulation of synovial fibroblasts by calcium oxalate and monosodium urate monohydrate. A mechanism of connective tissue degradation in oxalosis and gout. AB - The responses of cultured rabbit synovial fibroblasts to amorphous and microcrystalline calcium oxalate were compared with responses to MSUM. Like urate crystals, crystalline calcium oxalate (but not amorphous oxalate) caused marked stimulation of secretion of latent collagenase and PGE2 after 3 days of culture without significant change in cell protein or gross cellular morphology. Collagenase rose from undetectable levels in control cultures to 32.4 +/- 6.0 and 27.4 +/- 7.9 U/mg of cell protein for crystalline calcium oxalate and MSUM, respectively. PGE2 rose from a control level of 0.24 +/- 0.14 to 19.47 +/- 5.15 and 23 +/- 4.84 micrograms/mg of cell protein for crystalline calcium oxalate and sodium urate compared to 1.22 +/- 0.48 microgram for amorphous calcium oxalate. Although the crystalline species studied caused LDH in the media to increase threefold, this was minimal. Cell stimulation by amorphous oxalate and the crystals did not correlate with membranolytic potential as measured with an erythrocyte lysis assay. Stimulation of resident synovial cells by crystalline calcium oxalate and sodium urate may contribute to the chronic inflammation and destruction of joint tissues that occurs in oxalosis and gout. PMID- 6292316 TI - The (--)[3H]dihydroalprenolol binding to rat adipocyte membranes: an explanation of curvilinear Scatchard plots and implications for quantitation of beta adrenergic sites. AB - In rat adipocyte membranes, both beta-adrenergic agonists and beta-adrenergic antagonists competed with (--)[3H]dihydroalprenolol for high affinity (KD 2-4 nM) and low capacity binding sites. The antagonists but not the agonists competed with (--)[3H]dihydroalprenolol for lower affinity and higher capacity sites. The present studies were performed in order to characterize the adipocyte beta adrenergic receptor and distinguish it from low affinity, higher capacity sites which were heat-labile and not stereoselective. When isoproterenol was used to define the nonspecific binding, saturation studies showed a single binding site with a capacity of approximately 100 fmol/mg membrane protein (corresponding to approximately 50,000 sites/adipocyte). Binding was saturated by 10 nM (- )[3H]dihydroalprenolol. Approximate KD's of 204 nM were observed. Kinetic analysis of (--)[3H]dihydroalprenolol binding provided an independent measurement of KD between 0.75 and 1.1 nM. This binding site had the characteristics of a beta 1-adrenergic receptor with the potency of isoproterenol greater than norepinephrine greater than or equal to epinephrine as competitors of binding. Furthermore, the KD of inhibition of (--)[3H]dihydroalprenolol binding correlated with the Ki of inhibition by antagonists or Ka of activation by agonists of glycerol release in isolated adipocytes (r = 0.968, P less than 0.001). These results suggest that beta-adrenergic agonists compete with (- )[3H]dihydroalprenolol for the high affinity binding site which represents the physiological site. Furthermore, the use of antagonists (propranolol, alprenolol) to define specific beta-binding includes nonspecific site(s) as well as the beta adrenergic site. Previous characterization and quantitation of beta receptors in rat fat cell membranes may have been in error by incorporating both types of binding in their measurement. PMID- 6292317 TI - Receptor-mediated uptake of hypertriglyceridemic very low density lipoproteins by normal human fibroblasts. AB - Our previous studies showed that very low density lipoproteins, Sf 60-400 (VLDL), from hypertriglyceridemia subjects, but not VLDL from normolipemic subjects, suppress HMG-CoA reductase activity in normal human fibroblasts. To determine if this functional abnormality of hypertriglyceridemic VLDL resulted from differences in uptake of the VLDL by the low density lipoprotein (LDL) receptor pathway, we isolated VLDL subclasses from the d less than 1.006 g/ml fraction of normal and hypertriglyceridemic plasma by flotation through a discontinuous salt gradient for direct and competitive binding studies in cultured human fibroblasts. VLDL from the plasma of subjects with hypertriglyceridemia types 4 and 5 were at least as effective as normal LDL in competing for 125I-labeled LDL binding, uptake, and degradation when compared either on the basis of protein content or on a particle basis. By contrast, normolipemic Sf 60-400 VLDL were ineffective in competing with the degradation of 125I-labeled LDL, and Sf 20-60 VLDL (VLDL3) were less effective in reducing specific 125I-labeled LDL degradation than were LDL, consistent with their effects on HMG-CoA reductase activity. In direct binding studies, radiolabeled VLDL from hypertriglyceridemic but not normolipemic subjects were bound, internalized, and degraded with high affinity and specificity by normal fibroblasts. Uptake and degradation of iodinated hypertriglyceridemic VLDL Sf 100-400 showed a saturable dependence on VLDL concentration. Specific degradation plateaued at approximately 25 micrograms VLDL protein/ml, with a half maximal value at 6 micrograms/ml. The most effective competitor of hypertriglyceridemic VLDL uptake and degradation was hypertriglyceridemic VLDL itself. LDL were effective only at high concentrations. Uptake of normal VLDL by normal cells was a linear rather than saturable function of VLDL concentration. By contrast, cellular uptake of the smaller normal VLDL3 was greater than uptake of larger VLDL and showed saturation dependence. After incubation of normal VLDL with 125I-labeled apoprotein E, reisolated 125I-E-VLDL were as effective as LDL in suppression of HMG-CoA reductase activity, suggesting that apoE is involved in receptor-mediated uptake of large suppressive VLDL. We conclude that 1) hypertriglyceridemic VLDL Sf 60-400 are bound, internalized, and degraded by normal fibroblasts primarily by the high affinity LDL receptor mediated pathway; 2) by contrast, normal VLDL, Sf 60-400 are bound, internalized, and degraded by normal fibroblasts primarily by nonspecific, nonsaturable routes; and 3) of the normal VLDL subclasses, only the smallest Sf 20-60 fraction is bound and internalized via the LDL pathway. PMID- 6292318 TI - Decomposing potassium peroxychromate produces hydroxyl radical (.OH) that can peroxidize the unsaturated fatty acids of phospholipid dispersions. AB - The unsaturated fatty acyl residues of egg yolk lecithin are selectively removed when bilayer dispersions of the lipid are exposed to decomposing peroxychromate at pH 7.6 or pH 9.0. Mannitol (50 mM or 100 mM)partially prevents the oxidation of the phospholipid due to decomposing peroxychromate at pH 7.6 and the amount of lipid lost is inversely proportional to the concentration of mannitol. N,N Dimethyl-p-nitrosoaniline, mixed with the lipid in a molar ratio of 1.3:1, completely prevents the oxidation of lipid due to decomposing peroxychromate at pH 9.0, but some linoleic acid is lost if the incubation is done at pH 7.6. If the concentration of this quench reagent is reduced tenfold, oxidation of linoleic acid by decomposing peroxychromate at pH 9.0 is observed. Hydrogen peroxide is capable of oxidizing the unsaturated fatty acids of lecithin dispersions. Catalase or boiled catalase (2 mg/ml) protects the lipid from oxidation due to decomposing peroxychromate at pH 7.6 to approximately the same extent, but their protective effect is believed to be due to the non-specific removal of .OH. It is concluded that .OH is the species responsible for the lipid oxidation caused by decomposing peroxychromate. This is consistent with the observed bleaching of N,N-dimethyl-p-nitrosoanaline and the formation of a characteristic paramagnetic .OH adduct of the spin trap, 5,5-dimethylpyrroline-1 oxide. PMID- 6292319 TI - [Angioscintigraphy in venous pathology. Comparison of isotope angioscintigraphy and radiologic examination]. AB - Results of vascular radiology and isotope examinations were compared in 48 patients with suspected ileocaval lesions (37 cases) or affections of the veins of the upper limb or superior vena cava (11 cases). Isotopic examination, which can be performed in ambulatory patients, respects normal hemodynamic conditions, is painless, and can be repeated, was found to give valid results, positive correlations with radiological investigation findings being present in 43 cases. Lack of correlation in the remaining 5 patients could have resulted from the period of time elapsed between the two examinations or the techniques employed during each investigation. PMID- 6292320 TI - Anti-HAV in Thai population. PMID- 6292322 TI - The newest cephalosporins: how to use them? PMID- 6292321 TI - Effects of tetrodotoxin on electrical and mechanical activity of cardiac Purkinje fibers. AB - Canine Purkinje and ventricular muscle fibers were superfused in vitro and the actions of tetrodotoxin (TTX, 1.56 X 10(-5) - 3.2 X 10(-7) M) on both electrical and mechanical events were studied. Tetrodotoxin reduced the amplitude of the upstroke, markedly shortened the plateau, reduced the maximum diastolic potential and decreased contractile force in Purkinje fibers. Citrate (present in the TTX solution) did not have any effect on electrical and mechanical events. The modifications induced by TTX occurred whether external calcium concentration was normal, high or low. Furthermore, high calcium increased force by a similar extent whether TTX was present or not. TTX has little effect on slow action potentials due to the activation of the slow channel. Action potential duration and force were affected similarly by TTX as a function of concentration, of time of exposure and of rate of discharge. Also, TTX abolished action potentials due to a partially activated fast sodium current and reduced action potential duration and force much less in ventricular muscle than in Purkinje fibers. It is concluded that the effects of TTX are primarily related to a reduction of sodium influx during the action potential and that the fall in force is an indirect consequence of reduced sodium influx. PMID- 6292323 TI - Characterization of the purified principles of Formosan snake venoms which affect blood coagulation and platelet aggregation. PMID- 6292324 TI - Effects of a crotamine-like toxin from Crotalus h. horridus venom on skeletal neuromuscular transmission. PMID- 6292326 TI - Effects of valinomycin and MnCl2 on the ventricular fibrillation threshold and on aconitine-induced arrhythmia. PMID- 6292325 TI - Expression of Epstein-Barr virus genome in nude-mouse-growth nasopharyngeal carcinoma cells. PMID- 6292327 TI - Diagnostic value of Tc-99m DEIDA in hepatocellular carcinoma. PMID- 6292328 TI - Transmembrane Na+ and Ca2+ electrochemical gradients in cardiac muscle and their relationship to force development. AB - Na+- and CA2+-sensitive microelectrodes were used to measure intracellular Na+ and Ca2+ activities (alpha iCa) of sheep ventricular muscle and Purkinje strands to study the interrelationship between Na+ and Ca2+ electrochemical gradients (delta muNa and delta muCa) under various conditions. In ventricular muscle, alpha iNa was 6.4 +/- 1.2 mM and alpha iCa was 87 +/- 20 nM ([Ca/+] = 272 nM). A graded decrease of external Na+ activity (alpha oNa) resulted in decrease of alpha iNa, and increase of alpha iCa. There was increase of twitch tension in low alpha oNa solutions, and occasional increase of resting tension in 40% alpha oNa. Increase of external Ca2+ (alpha oCa) resulted in increase of alpha iCa and decrease of alpha iNa. Decrease of alpha oCa resulted in decrease of alpha iCa and increase of alpha iNa. The apparent resting Na-Ca energy ratio (delta muCa/delta muNa) was between 2.43 and 2.63. When the membrane potential (Vm) was depolarized by 50 mM K+ in ventricular muscle, Vm depolarized by 50 mV, alpha iNa decreased, and alpha iCa increased, with the development of a contracture. The apparent energy coupling ratio did not change with depolarization. 5 x 10(-6) M ouabain induced a large increase in alpha iNa ad alpha iCa, accompanied by an increase in twitch and resting tension. Under the conditions we have studied, delta muNa and delta muCa appeared to be coupled and n was nearly constant at 2.5, as would be expected if the Na-Ca exchange system was able to set the steady level of alpha iCa. Tension threshold was about 230 nM alpha iCa. The magnitude of twitch tension was directly related to alpha iCa. PMID- 6292329 TI - Control of intracellular pH. Predominant role of oxidative metabolism, not proton transport, in the eukaryotic microorganism Neurospora. AB - Recessed-tip microelectrodes were used to measure internal pH (pHi) in the fungus Neurospora, and to examine the response of pHi to several kinds of stress: changes of extracellular pH (pHo), inhibition of the principal proton pump in the plasma membrane, and inhibition of respiration. Under control conditions, at pHo = 5.8, pHi in Neurospora is 7.19 +/- 0.04. Changes of pHo between 3.9 and 9.3 affect pHi linearly but with a slope of only approximately 0.1 unit pHi per unit pHo, stable pHi being reached within 3 min of changed pHo. Despite a postulated high passive permeability of the Neurospora membrane to protons (Slayman, 1970), neither active nor passive H+ transport appears critical to pHi because (alpha) specific inhibition of the proton pump by orthovanadate has little effect on pHi, and (b) cytoplasmic acidification produced by respiratory blockade is unaffected by the size or direction of proton gradient. To convert measured changes in pHi into net proton fluxes, intracellular buffering capacity (beta i) was measured by the weak acid/weak base technique. At pHi = 7.2, beta i was (-) 35 mmol H+ (liter cell water)-1 (pH unit)-1, but beta i increased substantially in both the acid and alkaline directions, which suggests that amino acid side chains are the principal source of buffer. PMID- 6292330 TI - Monazomycin-induced single channels. I. Characterization of the elementary conductance events. AB - Monazomycin (a positively charged, polyene-like antibiotic) induces voltage dependent conductance changes in lipid bilayer membranes when added to one of the bathing solutions. These conductance changes have generally been attributed to the existence of channels spanning the membrane. In this article we characterize the behavior of the individual conductance events observed when adding small amounts of monazomycin to one side of a lipid bilayer. We find that there are several apparent channel types with one or sometimes two amplitudes predominating. We find further that these fairly similar amplitudes represent two different states of the same fundamental channel entity, presumed to be the monazomycin channel. The current-voltage characteristics of these channels are weakly hyperbolic functions of applied potential. The average lifetimes are essentially voltage independent (between 50 and 400 mV). The average channel intervals, on the other hand, can be strongly voltage dependent, and we can show that the time-averaged conductance of a membrane is proportional to the average channel frequency. PMID- 6292331 TI - Monazomycin-induced single channels. II. Origin of the voltage dependence of the macroscopic conductance. AB - The voltage dependence of the conductance induced induced in thin lipid membranes by monazomycin is shown here to be caused by voltage-dependent variations in the frequency of channel openings. We also experimentally demonstrate certain interesting properties of the channel activity that are predicted by a chemical kinetic model (Muller and Peskin, 1981), which successfully describes the macroscopic conductance. We conclude that two parallel mechanisms--one autocatalytic, the other simple mass action--exist that allow monazomycin to enter (or leave) the membrane so that the monazomycin molecules can be in a position to form channels. PMID- 6292332 TI - The Na-K pump as a current source. Affinity-driven transport has no reversal potential but its metabolic cost does. PMID- 6292333 TI - Functional stoichiometry at the nicotinic receptor. The photon cross section for phase 1 corresponds to two bis-Q molecules per channel. AB - These experiments examine changes in the agonist-induced conductance that occur when the agonist-receptor complex is perturbed. Voltage-clamped Electrophorus electroplaques are exposed to the photoisomerizable agonist trans-Bis-Q. A 1 microsecond laser flash photoisomerizes some trans-Bis-Q molecules bound to receptors; because the cis configuration is not an agonist, receptor channels close within a few hundred microseconds. This effect is called phase 1. We compare (a) the fraction of channels that close during phase 1 with (b) the fraction of trans-Bis-Q molecules that undergo trans leads to cis photoisomerization. Parameter a is measured as the fractional diminution in voltage-clamp currents during phase 1. Parameter b is measured by changes in the optical spectra of Bis-Q solutions caused by flashes. At low flash intensities, a is twice b, which shows that the channel can be closed by photoisomerizing either of two bound agonist molecules. Conventional dose-response studies with trans-Bis Q also give a Hill coefficient of two. As a partial control for changes in the photochemistry caused by binding of Bis-Q to receptors, spectral measurements are performed on the photoisomerizable agonist QBr, covalently bound to solubilized acetylcholine receptors from Torpedo. The bound and free agonist molecules have the same photoisomerization properties. These results verify the concept that the open state of the acetylcholine receptor channel is much more likely to be associated with the presence of two bound agonist molecules than with a single such molecule. PMID- 6292334 TI - Ca2+-dependent changes in cyclic GMP levels are not correlated with opening and closing of the light-dependent permeability of toad photoreceptors. AB - We have measured the levels of 3',5'-guanosine monophosphate (cyclic GMP) in isolated retinas from toad to investigate their correlation to the opening and closing of the light-dependent permeability of photoreceptors. When Ca2+-induced changes in cyclic GMP concentration are compared with the Ca2+-induced changes in the permeability of photoreceptor light-dependent channel, four quantitative dissimilarities are noted. First, when extracellular Ca2+ ([Ca2+]o) is reduced from normal physiological levels to between 10(-6) and 10(-7) M, the light dependent permeability is increased, but cyclic GMP levels are not significantly changed. Second, when [Ca2+]o is increased from 1.8 to 20 mM, the light-dependent permeability is suppressed, but cyclic GMP levels are decreased by only 10-15%, about one-quarter the decrease that can be obtained with bright illumination. Third, when [Ca2+]o is increased from 10(-8) M to 20 mM, the light-dependent permeability is closed rapidly, but the cyclic GMP decrease is slow. Fourth, when [Ca2+]o is lowered to 10(-8) M, the sensitivity of the light-dependent permeability to steady illumination is decreased by three to four orders of magnitude, but the sensitivity of the light-dependent decrease in cyclic GMP is not significantly affected. These observations indicate that there is no simple correlation between cyclic GMP levels and the permeability of the light-dependent channels and that Ca2+ can affect the conductance in the absence of changes in cyclic GMP content. PMID- 6292335 TI - Mutant strains of Salmonella typhimurium with defective phosphoribosylpyrophosphate synthetase activity. AB - A mutant of Salmonella typhimurium with undetectable phosphoribosylpyrophosphate (PRPP) synthetase activity in vitro and abnormally low PRPP pools in vivo was identified by screening temperature-sensitive isolates by an autoradiographic procedure. The lack of PRPP synthetase activity in vitro and temperature sensitive growth were shown to result from separate, but closely linked mutations mapping at 47 units on the Salmonella chromosome. Mutant cell extracts prepared by a variety of methods did not show any detectable PRPP synthetase activity, but material that was immunochemically cross-reactive with PRPP synthetase was detected by complement fixation analysis. A second mutant, isolated by localized mutagenesis, contained about half the PRPP synthetase and cross-reacting material of the parental strain. PMID- 6292336 TI - Mouse mammary tumour virus and polyoma virus information in mammary tumours of athymic mice inoculated with polyoma virus. AB - Polyoma virus inoculation of athymic mice results in the development of mammary tumours with a much higher incidence than the development of salivary gland tumours, the latter being the most common for immunocompetent normal mice. The possibility existed that polyoma virus might act as a co-carcinogen in activating the expression of mouse mammary tumour virus (MMTV). Molecular hybridization studies, however, showed that the mammary tumour development was accompanied by neither the amplification of MMTV genomic sequences nor by their more extensive transcription. In contrast, tumour tissue contained about 60 to 100 copies of polyoma virus genome equivalents per cell and some of these sequences were apparently transcribed into RNA. While these results do not rule out the transient involvement of MMTV expression in mammary tumour development, it appeared that the mammary gland cells were directly transformed by polyoma virus. Apparently, polyoma virus displayed a tropism in athymic mice that was different from that in normal mice. PMID- 6292337 TI - Characterization of xenotropic and dual-tropic type C retroviruses isolated from Abelson tumour. AB - Tumours induced in Balb/c mice by Abelson virus complex were found to contain a xenotropic virus (A-X-MuLV) and an NB-tropic, dual-tropic virus (NBX), in addition to the Moloney leukaemia virus (M-MuLV) and the defective, transforming Abelson virus genome. Both A-X-MuLV and NBX virus were presumably present in a genomically masked form and could be recovered only by co-cultivation of tumour cells with permissive cells. Only about 0.87% and 0.13% of the viruses in the co culture supernatant represented A-X-MuLV and NBX virus respectively; the majority were M-MuLV. The NBX virus acted more efficiently than the HIX virus (Fischinger et al., 1975) as helper to rescue murine sarcoma virus (MSV) from S+L-cells of hamster, rat and mouse origin, whereas the converse was true for those of cat and human origin. The interference and nuetralization patterns suggested that the NBX virus was an env gene recombinant between A-X-MuLV and M-MuLV. The fact that NBX virus cross-reacted in radioimmunoassays with gp70s of both M-MuLV and Balb: virus-2 provides evidence for the recombinant nature of the NBX gp 70-coding gene which was probably derived from both M-MuLV and a virus similar to Balb: virus-2 or A-X-MuLV. The presence of a unique antigenic determinant on the gp70 of NBX virus is also suggested. Both A-X-MuLV and NBX virus cross-reacted with type specific p12s of M-MuLV and Balb: virus 2- suggesting that the gag gene coding for p12 of NBX virus was derived from the A-X-MuLV, which was itself a recombinant, its p12-coding gene being derived from both Balb: virus-2-like virus and M-MuLV. The NBX virus was not oncogenic when tested in newborn Balb/c mice. PMID- 6292338 TI - Correlation of the virus sequence content and biological properties of cells carrying the herpes simplex virus type 2 thymidine kinase gene. AB - Seven cell lines, transformed to a thymidine kinase-positive phenotype with herpes simplex virus type 2 (HSV-2) DNA fragments, have been examined to determine their virus-specific DNA sequence content. The results are consistent with the reported map position of the HSV-2 thymidine kinase gene. Different cell lines contained different amounts of non-selected virus-specific sequences and this correlated with the ability of some cell lines to compensate the deficiencies in some temperature-sensitive (ts) mutants of HSV-1 and HSV-2. The cell lines were also examined for their ability to synthesize elevated levels of thymidine kinase in response to infection with a thymidine kinase-negative virus mutant. Of the seven cell lines, two failed to respond to infection in this way and these cell lines lacked sequences on the upstream side of the kinase gene which were present in the remaining cell lines. PMID- 6292339 TI - Comparison of the oligosaccharide structure of the glycoprotein of vesicular stomatitis virus and a thermolabile mutant (tl-17). AB - As a means of examining the extent to which the polypeptide structure of a virus glycoprotein contributes to the overall structure and composition of the carbohydrate moieties, we have made a detailed comparison of the structure of the oligosaccharide moieties of wild-type vesicular stomatitis virus (VSV) glycoprotein with those of a glycoprotein-defective mutant of VSV, tl-17 (VSV). Characterization of the oligosaccharides by ion-exchange and gel filtration chromatography after sequential enzymic degradation reveals similar structures in the wt and mutant glycoproteins. However, the altered polypeptide structure of the tl-17 glycoprotein affects the extent of addition of sialic acid and fucose, both of which are added late in the maturation of the glycoprotein. PMID- 6292340 TI - Proteins specified by Herpesvirus saimiri: purification and properties of a single polypeptide which elicits virus-neutralizing antibody. AB - A virus-specified polypeptide of 160,000 mol. wt. (160K) was purified more than 1000-fold from the soluble proteins present in the culture medium of cells productively infected with herpesvirus saimiri (HVS). Purified preparations of the 160K polypeptide gave rise to high titre precipitating and neutralizing antibodies in immunized rabbits, which cross-neutralized independent isolates of HVS (neutralization rate constants, k, of between 3.8 and 4.8) and specifically precipitated the 160K polypeptide from extracts of cells infected with the homologous or heterologous strains of this virus. Antiserum to the 160K polypeptide of HVS also gave low (k = 0.02) levels of neutralization of the related herpesvirus ateles. Purified preparations of the 160K polypeptide were capable of removing most or all of the neutralizing antibody sera of squirrel monkeys with naturally acquired antibodies to HVS. The 160K polypeptide was previously shown to form part of the surface of enveloped virus particles. However, we show here that the 160K polypeptide is not extensively glycosylated in infected cells. The majority of glucosamine incorporation specific to HVS infected cells was into eight regions with apparent mol. wt. of 170K to 220K, 125K to 145K, 115K to 120K, 83K to 88K, 65K to 75K, 52K to 58K, 25K to 27K and 12.5K to 13K. It remains possible that alternative cleaved or glycosylated forms of the 160K polypeptide are also present on the virus particle or that precipitating antibody to 160K and virus-neutralizing antibodies are not identical. However, our inability to detect precipitating antibody to virus induced proteins or glycoproteins other than the 160K polypeptide and the high titre of neutralizing antibody present in this serum, provides reasonable evidence that it is the antibodies which react with the 160K polypeptide that are responsible for virus neutralization. PMID- 6292341 TI - Effects of n-butyrate on Epstein-Barr virus-carrying lymphoma lines. AB - n-Butyrate has been shown to induce Epstein-Barr virus (EBV) antigen synthesis in certain EBV-carrying lymphoma lines (Luka et al., 1979). We have studied the effect of n-butyrate on two EBV-positive Burkitt lymphoma lines by immuno fluorescence and electron microscopy. In the producer line P3HR-1, the drug induced not only early antigen (EA) and virus capsid antigen (VCA) synthesis, as shown before, but also increased the number of cells containing virus particles. The transition from VCA expression to the formation of virus particles was much more effective in treated cells than in EBV antigen-producing cells of the same line. The productive cycle was associated with the development of characteristic morphological changes. In the non-producer Raji cells, n-butyrate induced EA in only a minor fraction of the cells. There were, however, clear signs of differentiation in the direction of plasma cells. Two days after the addition of n-butyrate 80% of the Raji cells could be classified as plasmablasts. After 72 h, 20% of the cells appeared as typical plasma cells. PMID- 6292342 TI - Involvement of natural killer cells in the pathogenesis of murine cytomegalovirus interstitial pneumonitis and the immune response to infection. AB - The significance of the natural killer (NK) cell response to murine cytomegalovirus (MCMV) infection was evaluated in C3H/HeN mice. This strain was selected for study after preliminary demonstration that the NK cell response, occurring between 3 and 6 days post-infection was relatively high in comparison to other mouse strains studied. A dose-response effect of hydrocortisone treatment on suppression of this response was found. A dose of hydrocortisone, given subcutaneously on two successive days, which was found to markedly inhibit the NK cell response, had no effect on development of serum interferon or antibody levels, or spleen cytotoxic T cell activity under the conditions studied. Suppression of the NK cell response by this treatment, however, was accompanied by enhanced spleen and pulmonary virus replication in vivo and increased susceptibility of mice to lethal infection. MCMV interstitial pneumonitis was characterized histologically and lung lymphocytes studied at 4 days post-infection were found to have increased NK cell activity. Treatment of mice with hydrocortisone was found to inhibit development of gross and histological evidence of pneumonitis. These findings indicate that NK cells are involved in the pathogenesis of MCMV interstitial pneumonitis and may function early in infection to restrict the extent of virus replication. PMID- 6292343 TI - Condensed DNA structures derived from bacteriophage heads. AB - Bacteriophage lambda particles were rendered osmotically fragile by incubation, spread over hypophase and examined by electron microscopy. When water was used as hypophase, condensed structures were released from the phage heads and treatment of these with cytochrome c or several alternative proteins resulted in the release of free, relaxed DNA. Phage were pretreated with nitrogen mustard, a bifunctional alkylating agent; when the condensed structures from such phage particles were treated with protein, DNA was released in small supercoiled domains. This confirmed a previous finding that bacteriophage DNA has a supercoiled topology and suggests that the winding pattern of DNA in the phage might involve small domains of coiled DNA analogous to nucleosomes. Such a conformation could be consistent with other studies on the arrangement of DNA in phage heads if the domains have parallel axes. PMID- 6292344 TI - Nucleotide sequence of fowl plague virus RNA segment 7. AB - Nucleotide sequence analysis of a recombinant DNA clone of RNA segment 7 from FPV/Rostock/34 has shown it to be highly conserved in comparison with RNA segment 7 from two human strains (Allen et al., 1980; Winter & Fields, 1980; Lamb & Lai, 1981). FPV RNA segment 7 contains the coding capacity for two polypeptide chains. The sequence homology between RNA segment 7 of avian and human viruses was greater than 90%, and most of the changes did not result in amino acid substitutions. PMID- 6292345 TI - Identification of polypeptide precursors to HSV-1 glycoproteins by cell-free translation. AB - Using antisera of limited specificity we have detected among the in vitro translation products of HSV-1 RNA, polypeptides antigenically related to infected cell glycoproteins. The results obtained suggest that non-glycosylated polypeptide precursors of mol. wt. 85,000 and 52,000 correspond to infected cell glycoproteins of mol. wt. 120,000 to 126,000 and 56,000 to 68,000 respectively. PMID- 6292346 TI - Characterization of heavy particles of adeno-associated virus type 1. AB - The temperature-sensitive mutant ts4 of adenovirus type 2 (Ad-2) is capable of complementing adeno-associated virus type 1 (AAV-1) in HEp2, KB and HEK cells at 34 degrees C and 39 degrees C when used as a helper virus. Heavy non-infectious AAV-1 particles can be generated by using the mutant ts4 in HEp2 cells. When AAV 1 is grown in serial passages in HEp2 cells, both the wild-type Ad-2 and the mutant ts4 give rise to heavy, less infectious AAV-1 particles. The heavy AAV-1 particles generated by Ad-2 in advanced serial passages retain the property of having CF and IF antigens, but the AAV-1 generated by the mutant in advanced serial passages lose this property. There is no appreciable difference in the particle counts made by electron microscopy of AAV-1 preparations generated either by Ad-2 or the mutant ts4. Analysis by polyacrylamide gel electrophoresis of purified heavy AAV generated by ts4 indicates that in late passage an additional polypeptide of higher mol. wt. than the three structural polypeptides is detected. PMID- 6292347 TI - Restriction endonuclease mapping of unintegrated proviral DNA of Kirsten murine sarcoma virus. AB - Linear unintegrated Kirsten murine sarcoma virus DNA synthesized after acute infection of NIH/3T3 cells or by detergent-disrupted virions, was studied by restriction enzyme cleavage and agarose gel electrophoresis. Labelled DNA (cDNA) synthesized by reverse transcription of a virion RNA template was used to detect viral DNA sequences by filter hybridization. The sites of cleavage for eleven enzymes were located on the genome. Hybridization studies using a cDNA probe specific for the 3' end of the genome defined the orientation of the physical map with respect to virion RNA and identified terminally redundant sequences on the genome. Further evidence for terminal redundancy was obtained by restriction mapping of linear and circular viral DNA, where the duplication was shown to be 0.34 x 10(6) (500 base pairs) in length and in a tandem orientation. Circular viral DNA was found predominantly in the nucleus of acutely infected cells and it existed in two major size classes. The larger size class represented an exact circularization of linear DNA (with no other sequence permutation) whilst in the smaller DNA species one of the terminally redundant sequences was absent. PMID- 6292348 TI - Transfection with the isolated herpes simplex virus thymidine kinase genes. I. Minimal size of the active fragments from HSV-1 and HSV-2. AB - We have defined the minimal size and physical map locations in the genomes of both herpes simplex virus types 1 and 2 (HSV-1 and HSV-2) for DNA sequences capable of conferring stable biochemical transformation under thymidine kinase (TK) selection. The experiments involved transfection of Ltk- cells with either isolated virus DNA fragments or cloned pBR322 plasmids containing the 3.5 kilobase (kb) BamHI-O fragment from HSV-1(MP) or the 5.6 kb SalI-G fragment from HSV-2(333). Mapping of restriction enzyme sites within these cloned DNAs, followed by assays for colony formation in HAT medium after transfection with cleaved DNA, localized the biologically active TK-transforming sequences to lie between coordinates 0.300 and 0.313 in HSV-1 and between 0.303 and 0.315 in HSV 2. Experiments with a series of cloned plasmids containing deletions of the BamHI O fragment towards either the 3'- or 5'-ends of the TK gene indicated that the sequences required for stable HSV-1 TK transformation lay within a 1600 base pair (bp) region at 0.303 to 0.313 map units. An internal deletion mutant plasmid, selected by a novel bacterial transfection assay for the absence of the KpnI site at 0.308, also failed to rescue Ltk- cells. With the exception of cleavage at the StuI site at 0.303 in HSV-2, which reduced activity only eightfold, all cleavages that affected TK transformation reduced the efficiency at least 50-fold. A direct comparison of the HSV-1 and HSV-2 minimal transforming regions with the nucleotide sequence of the structural HSV-1 TK gene indicates that the HSV-2 StuI site lies 30 bp beyond the poly(A) addition site at the 3'-end of TK mRNA. On the other hand, cleavage at the SmaI site in HSV-1 TK, located 80 bp in front of the poly(A) addition point, abolishes colony formation. Comparison of the putative 5' end of the HSV-2 TK gene defined by transfection assays, with a 250 bp non transcribed region at the front of the HSV-1 TK gene, suggests that the promoter regions contain a much higher frequency of conserved cleavage sites than do the coding portions of the two genes. Direct nucleotide sequencing of the 5'-flanking sequences for HSV-2 TK confirmed that large portions of the two promoters possess greater than 95% sequence homology. At least 140 bp, but no more than 200 bp, of this 5'-promoter region are essential for efficient transfer and expression of the viral TK gene. Combining the results from HSV-1 and HSV-2, we conclude that a contiguous sequence of 1480 to 1540 bp is necessary to achieve at least 10% of the maximum transformation efficiency. PMID- 6292349 TI - The molecular biology of Yaba tumour pox virus: analysis of lipids, proteins and DNA. AB - Cytopathological studies have shown that Yaba tumour pox virus (Yaba virus) infection leads to the accumulation of large lipid vacuoles. The rate of accumulation of these vacuoles increased as the infection proceeded. These lipid vacuoles were not seen in control cells or in cells infected with monkeypox virus (MPV) but were seen during Yaba virus infection in the presence of cytosine arabinofuranoside (100 microgram/ml). Yaba virus also failed to inhibit host protein synthesis as infection proceeded for prolonged periods. Yaba virus proteins were shown to be substantially different from those of MPV when analysed by two-dimensional electrophoresis. The genome of Yaba virus gave restriction enzyme fragments which differed from those of the MPV genome when cleaved with the enzymes HindIII and XhoI. However, Yaba virus DNA hybridized to the HindIII fragments K, L and M and to the XhoI fragments A, B, C, E and G of MPV DNA. PMID- 6292350 TI - Transfer of murine leukaemia and murine sarcoma virus genetic information by transfection with isolated metaphase chromosomes. AB - Chromosome-mediated transfer of murine leukaemia (MuLV) and murine sarcoma (MuSV) virus genetic information to uninfected recipient cells was investigated. Metaphase chromosomes from AKR MuLV-infected SC-1 mouse cells were incubated with NIH/3T3 cells. After several passages (1 to 3 weeks), infectious virions exhibiting reverse transcriptase activity and the characteristic host range of ecotropic, N-tropic AKR virus appeared in the supernatant fluids of the treated cells. Restriction endonuclease analysis of genomic DNA from transfected cells indicated that AKR proviral DNA was associated with the high molecular weight DNA of the host. These results demonstrate that the AKR MuLV genome can be stably transferred to uninfected recipient cells via isolated metaphase chromosomes. Although AKR virions are not able to infect heterologous cells, chromosome mediated transfection resulted in the establishment of productive AKR MuLV infection in mink cells. Thus, the use of chromosomes to transfer virus genes can circumvent the natural host restriction barrier. In other experiments, it was shown that normal NIH/3T3 cells were transformed after exposure to metaphase chromosomes isolated from an MuSV-infected, non-producer line. Foci were detected 14 to 21 days after chromosome treatment and were shown to contain true viral transformants since transforming virus was produced after superinfection with MuLV. PMID- 6292351 TI - Molecular epidemiology of tick-borne encephalitis virus: peptide mapping of large non-structural proteins of European isolates and comparison with other flaviviruses. AB - Nine virus-specified proteins were identified by SDS-polyacrylamide gel electrophoresis in [35S]methionine-labelled chick embryo cells infected with tick borne encephalitis (TBE) virus by comparison with mock-infected cells. These proteins were designated P91, p74, p72, P67, GP53(E), P47, p25, P15(C) and P14.5 according to their molecular weights. Peptide mapping of P91, P67, GP53(E) and P47 from TBE virus-infected cells, as well as those of the corresponding proteins from West Nile virus (WNV)-infected cells (previously termed NV5, NV4, V3 and NV3), demonstrated the uniqueness of these proteins. Almost no subtype variability, with respect to the pattern of intracellular proteins, was found when isolates of TBE virus from Austria, Switzerland, Germany, Finland and Czechoslovakia were compared. Peptide mapping of NV5 (P91) and NV4 (P67) from all these isolates using limited proteolysis with alpha-chymotrypsin and V8 protease revealed completely identical patterns, thus extending our observations that TBE virus seems to represent a very stable member of the flavivirus genus, which was based on the lack of variation found with the structural glycoprotein. On the other hand, the Far Eastern subtype of TBE virus and the closely related louping ill virus could not only be differentiated from the Western subtype by differences in the peptide maps of their structural glycoprotein but also in those of the non-structural protein NV5, i.e. subtype or subgroup variations are not confined to the virion surface glycoprotein. WNV and Murray Valley encephalitis virus (MVEV) revealed the expected heterogeneity of virus-specified proteins found in cells infected with different flaviviruses. It is especially interesting that also the largest non-structural protein, NV5, is subject to this heterogeneity, ranging in mol. wt. from 91 000 for TBE virus to 98 000 for MVEV and that also the peptide maps of NV5, as well as those of NV4, were unrelated. These proteins, therefore, revealed a variability between serologically distinct flaviviruses similar to that observed with the structural glycoprotein. PMID- 6292352 TI - Neutralization of foot-and-mouth disease virus. II. Further parameters related to the sensitization of the 140S virion by antibody. AB - The reaction of foot-and-mouth disease virus (FMDV) with 12S subunit/140S virion cross-reactive (sensitizing) antibody was studied in order to elucidate the requirements for neutralization versus sensitization. The presence of sensitizing antibody in immune serum caused an atypical in vitro neutralization response curve and a non-neutralized fraction. Cell-associated (cytophilic) antibody was not present in the system. Dissociation of the immune complex was not a factor and sensitized virus adsorbed to host cells via the regular virus receptor site(s). This finding led to the conclusion that sensitizing antibody is specific for non-critical sites. Dosing of the neutralization reaction mixtures with fractionated antibody of alternative antigenic specificities had an antagonistic effect on the neutralization response, suggesting steric hindrance. Cell receptor sites were a factor in sensitization since different host systems had different susceptibilities for sensitized antigen. The results suggest that in vitro neutralization of FMDV requires the attachment of multiple antibody molecules as proposed by the multi-hit theory of neutralization. The in vitro measurement of serum neutralizing activity as an indication of the in vivo immune response is discussed. PMID- 6292353 TI - Identification of a neutralization-specific antigen of a calf rotavirus. AB - Monospecific polyclonal antisera were raised in guinea-pigs against the calf rotavirus polypeptides VP1, VP2, VP3 + 4, VP4.2, VP6, VP7.1, VP7.2 and VP10. All of the antisera gave a similar pattern of cytoplasmic immunofluorescence in rotavirus-infected cells, but spots of fluorescence of varying intensity with different sera were seen over the nucleus. Immune precipitation, using Staphylococcus aureus to collect immune complexes, showed that VP2 was precipitated by antiserum to VP2 (alpha-VP2) and VP6 by alpha-VP6, alpha-VP7.1 and alpha-VP7.2 both precipitated the same range of proteins from infected cells (VP7, VP7.1 and VP7.2) or from virions (VP7.1 and VP7.2). VP10, either from virions or infected cells, was not precipitated by alpha-VP10. The only antiserum which efficiently neutralized infectivity was alpha-VP7.2. There were low levels of neutralization with alpha-VP10 (but the results varied from experiment to experiment) and traces with alpha-VP6. alpha-VP7.1 and the other antisera did not neutralize even though alpha-VP7.1 agglutinated double-shelled particles as seen in immune electron microscopy to a greater extent than alpha-VP7.2. Both VP7.1 and VP7.2 were shown to be glycoproteins by tunicamycin treatment of infected cells. Core particles only were agglutinated by alpha-VP10. All the evidence leads us to conclude that there were major neutralizing antigenic determinants present on VP7.2, a minor component of the outer shell of the virion. PMID- 6292354 TI - Coliphage BA14: a new relative of phage T7. AB - Coliphage BA14 was isolated from sewage and shown to be related to phages T7 and T3. It is similar to T3 in that it directs the synthesis of an S-adenosyl methionine-cleaving enzyme (SAMase) early upon infection. However, it differs from all other known T7-related coliphages by the inability of its RNA polymerase (gene 1 product) to transcribe T7 DNA or T3 DNA. BA14, T7 and T3 also show marked differences in autoradiographic patterns of their gel-electrophoretically separated 35S-labelled intracellular phage proteins, restriction endonuclease HpaI cleavage patterns of their DNAs, and serological specificities of their infectious particles. Other distinctive features became apparent upon simultaneous mixed infection with BA14 and T7 or T3: inability of BA14 to produce genetic recombinants with either T7 or T3; lack of functional complementation between amber mutants of BA14 and T7 or T3; mutual exclusion and depression of the burst size of the mixedly infected cells. PMID- 6292355 TI - The positively charged structural virus protein (VP1) of foot-and-mouth disease virus (type O1) contains a highly basic part which may be involved in early virus cell interaction. AB - Polypeptides of 'trypsin-resistant' (TR) variants of foot-and-mouth disease virus type O1 (BFS 1860) were analysed by electrofocusing and two-dimensional gel electrophoresis. In contrast to parent O1 virus, trypsin treatment of these variants did not reduce their infectivity and their ability to attach to susceptible cells, although VP1 was cleaved as in the parent virus. In OTR1, one of the cloned isolates, an additional polypeptide (VPA) with a mol. wt. approx. 31 X 10(3) (31K), was found which resembled VP1 (28K) in being positively charged and cleaved by trypsinization of the virus into a neutral 18K polypeptide (P18) and a strongly basic fragment (pI greater than 10) with a mol. wt. of approx. 6K (P6). These findings substantiate the hypothesis that VPA is an elongated VP1. While P18 fragments of both trypsin-treated parent virus and OTR progeny viruses focused at identical (neutral) pH, P6 fragments of trypsinized OTR variants (including OTR1) were even more positively charged than P6 of parent virus. This difference in charge of the P6 polypeptide may be responsible for the retained cell attachment ability of trypsinized OTR viruses. The data are discussed with respect to the known amino acid sequence of VP1 of the closely related O1 Kaufbeuren. PMID- 6292356 TI - A mutant standard virus isolated from vesicular stomatitis virus persistent infection interferes specifically with wild-type virus replication. AB - A clone of vesicular stomatitis virus (VSV) isolated after 76 months of persistence exerts specific homologous interference with replication of wild-type or tsG31 (temperature-sensitive) virus in a mixed infection at 37 degrees C in the absence of defective-interfering particles. Other VSV ts mutants showed little or no specific interfering ability. PMID- 6292357 TI - Monoclonal antibody specific for avian sarcoma virus structural protein p27. AB - A hybridoma cell line which secretes antibody to the Rous sarcoma virus (RSV) structural protein p27 has been established. The hybrid resulted from the fusion of NS-1 myeloma cells with spleen cells from a Balb/c mouse which was immunized with RSV-transformed mouse cells. Antibodies produced by the hybrid clone immunoprecipitated p27 and gag precursor proteins (Pr180gag,pol, Pr76gag and Pr66gag) from [35S]methionine-labelled chicken embryo fibroblasts transformed by the Schmidt-Ruppin strain of RSV. When Schmidt-Ruppin virus was radioactively labelled with [35S]methionine, p27 was the only virus structural protein immunoprecipitated. Antibody production by the hybrid clone (designated 7-29-D6) has remained stable for longer than 12 months at a level of 50 micrograms IgG/ml medium. A highly sensitive method to determine the subclass specificity of monoclonal antibodies is described. In this procedure, the clone is incubated with [35S]-methionine, and radiolabelled antibody is precipitated with affinity purified, subclass-specific rabbit anti-mouse serum and Staphylococcus aureus. The advantages of this procedure are discussed. PMID- 6292358 TI - Low molecular weight IgM in cytomegalovirus infections and cardiac patients. AB - When 71 ischaemic heart disease and cardiomyopathy patients' sera were examined by the IFA (indirect fluorescent antibody) test, 18 of the 49 (36%) with CMV (cytomegalovirus) CF antibody appeared to contain a specific LMW (7S approx) IGM. This IgM was unrelated to the presence of RF (rheumatoid factor) demonstrable by agglutination of IgG-coated latex and by an immunofluorescence test, was not absorbed by heat-aggregated IgG and latex, nor did its demonstration appear to be complement dependent. It was demonstrable in acute CMV infection and in some cardiac patients, binds to HSV (herpes simplex virus) and CMV-infected cells. PMID- 6292359 TI - Neutralization and sensitization of cytomegalovirus by IgG antibody, anti-IgG antibody, and complement. AB - Human cytomegalovirus (CMV) strain Ad 169 was reacted with IgG antibody obtained from infected renal transplant patients, and the degree of neutralization was determined. The mixture of antibody and virus was then incubated with anti-human IgG (A-IgG) or complement (C) to measure additional neutralization by these agents and thus to estimate the concentration of infectious virus-antibody complexes which had been formed. Neutralization and formation of infectious complexes susceptible to neutralization by A-IgG or C decreased with lower concentrations of antibody. The rate of neutralization of preformed complexes by A-IgG or C was more rapid than the formation of complexes or the neutralization of native virus. If equally infectious suspensions of native virus and virus plus antibody were challenged with additional CMV antibody both solutions were neutralized to the same degree. The data are most compatible with the suggestion that the formation of infectious virus-antibody complexes is the initial step in the neutralization process. These complexes do not appear to be protected from further neutralization. PMID- 6292360 TI - Virus receptors for polymerized human albumin: a prognostic marker in HBeAg positive chronic hepatitis type B? AB - Seventeen out of 30 patients with chronic hepatitis type B with hepatitis B e antigen (HBeAg) in serum remained persistently positive for e antigen, while 13 seroconverted to antibody (anti-HBe) when followed over a period of one to five years. Initial levels of serum hepatitis B virus (HBV) markers, such as the hepatitis B surface antigen (HBsAg), HBeAg, and HBV-DNA polymerase (HBV-DNAP) were similar in the two groups of patients, while initial titres of the HBsAg associated receptor for polymerized human serum albumin (pHSA), recently identified on HBV particles, were significantly higher in the patients who remained HBeAg positive (mean titre +/- SD = 2(-7.00) +/- 2(-3.2)) compared to the cases who eventually seroconverted to anti-HBe during the follow-up (2(-2.54) +/- 2(-2.14) P less than 0.001). A receptor titre above 1:64 by haemagglutination was highly predictive of persistence of HBeAg, suggesting that in patients with HBeAg-positive chronic hepatitis testing for the HBsAg-associated pHSA receptor may be useful in predicting the duration of HBe antigenaemia, with relevant clinical and prognostic implications. PMID- 6292361 TI - The role of BK virus in acute respiratory tract disease and the presence of BKV DNA in tonsils. AB - The significance of BKV infections relative to infections by generally tested respiratory agents was investigated in children with acute respiratory disease. Paired sera from 177 children admitted to a hospital for acute respiratory disease were tested for significant rises in antibodies. Sera from seven patients showed a seroconversion to BKV and clinical signs of acute upper respiratory tract infection were exhibited by each of these patients. BKV infections were present in 8% of the patients with upper respiratory tract disease while seroconversions to adenovirus (2%), influenza A virus (1%), parainfluenza virus (5%), RS virus (6%) and mycoplasma pneumoniae (1%) were observed in 15% of the patients with upper respiratory tract disease. BKV was isolated from the urine of one child with tonsillitis with a concomitant seroconversion to BKV. Tonsils from children with recurrent attacks of acute respiratory disease were tested for the presence of BKV DNA by hybridization with a cloned genomic 32P-labeled DNA of prototype BKV. Five of twelve tonsil DNAs showed hybridization with BKV DNA. Each tonsil showing hybridization with BKV DNA contained multiple nonintegrated copies of the BKV genome per diploid amount of host cell DNA. Attempts to recover infective BKV by transfection of primary human embryonic cells with tonsil DNAs or by co-cultivation of tonsillar cells with primary human embryonic cells were unsuccessful. PMID- 6292362 TI - Limbic ictus and atypical psychoses. PMID- 6292363 TI - The benzodiazepine/GABA receptor complex: molecular size in brain synaptic membranes and in solution. AB - The molecular size of the benzodiazepine (BZ) receptor in the synaptic membrane of brain cortex (bovine or rat) was determined by an improved version of the radiation inactivation method to be 220,000. An identical size was found simultaneously for the associated gamma-aminobutyric acid (GABA) receptor and for the component binding beta-carboline esters. It is proposed that all three activities reside in a single protein or protein complex in the membrane. The size in solution, after extraction into Triton X-100 medium from exhaustively washed membranes, was estimated by sedimentation constant (9.4S) and by gel filtration ( approximately 230,000 apparent MW), again with the BZ and GABA binding activities behaving identically. This size applies to the component that undergoes photoaffinity labelling by [3H]flunitrazepam in the membrane, and contains a 51,000 Mr polypeptide as the BZ-binding subunit. It is concluded that a protein complex or oligomer of 200,000-220,000 MW carries a class of BZ-binding sites and an associated class of GABAA sites. PMID- 6292364 TI - Solubilization by CHAPS detergent of barbiturate-enhanced benzodiazepine-GABA receptor complex. AB - Barbiturates enhance the binding of [3H]flunitrazepam to benzodiazepine receptors solubilized with the detergent 3-[(3-cholamidopropyl) dimethylammonio]propanesulfonate (CHAPS) from bovine cortex. The enhancement by the barbiturates is seen as a decrease in the dissociation constant, KD, for specific benzodiazepine binding, with no effect on the number of binding sites. The effect of the barbiturates is facilitated by chloride ions, is concentration dependent, and has a specificity that correlates well with the anesthetic potency of barbiturates. [3H]Flunitrazepam binding activity is stable with storage at 4 degrees C, but barbiturate enhancement of soluble benzodiazepine binding activity decayed rapidly (t 1/2 = 48 h). [3H]Muscimol binding (GABA receptor) activity was also enhanced by barbiturates. Agarose gel filtration column chromatography of the CHAPS-solubilized receptor proteins showed the same elution profile as receptors solubilized with sodium deoxycholate, and enhancement by barbiturates was observed for both the benzodiazepine and GABA binding activities. PMID- 6292365 TI - Rapid cellular regulation of D-glucose transport in cultured neural cells. AB - Previous studies have revealed two different kinds of regulation of glucose utilization in cell lines derived from the nervous system (Keller et al., 1981). We found glucose metabolism of C-6 glioma cells to be limited and regulated by membrane transport. In contrast, glucose utilization of C-1300 neuroblastoma (N2A) cells was limited by the known regulatory enzymes of the Embden-Meyerhof pathway. Under the given experimental conditions the "membrane-limited" C-6 glioma cells were characterized by periodically changing glucose transport rates and very low intracellular glucose concentrations, which remained constant in spite of widely differing transport rates. These findings suggest the close functional coupling between transport and phosphorylation required for the regulation of glucose transport by cellular metabolic needs. PMID- 6292366 TI - Opiate receptor-mediated inhibition of catecholamine release in primary cultures of bovine adrenal chromaffin cells. PMID- 6292367 TI - Factors contributing to the poor myelination in the brain of the Snell dwarf mouse. AB - Conventional histological examination of the pituitary does not distinguish Snell dwarf mutants (dw/dw) from their normal littermates (+/?) in the neonatal stage. However, immunohistochemical examination of pituitaries of litters born to heterozygous Snell parents revealed that in approximately 25% of the glands examined, the number of positive cells was very low in the neonatal stage. We attempted to delineate the events resulting in the poor myelination in the brain of the Snell dwarf mouse, and to devise an immunohistochemical method for identifying the mutant neonate. Differences in the brain weights of the dw/dw and +/? mice first became apparent on the 10th day of age, and from this time on no further increase in the weight of the dwarf mouse brain was recorded. Increase in CNPase activity was found to be suppressed in the cerebrum and brain stem throughout the developmental stage, but not in the other parts of the brain. The yield of isolated myelin decreased by 58% in the mutant mouse, but CNPase activity was equivalent to that of control myelin. Differences in DNA content per cerebrum from the dw/dw and +/? mice first became apparent on the 10th day of age. Henceforth, the dw/dw mice showed no further increase, although the +/? mice continued to increase. [3H]Thymidine incorporation into the DNA fraction in vivo on the 7th day of age, when glial cell proliferation in the cerebrum is most active, was suppressed to about 50% of the control level in all parts of the dwarf brain. These findings indicate that the poor myelination found in the mutant cerebrum is a hypomyelination due to reduce oligodendroglial proliferation caused by lack of circulating growth hormone. PMID- 6292368 TI - An improved filtration procedure for measuring opiate receptors in small regions of rat brain. AB - A modified filtration method for in vitro receptor binding was used to determine specific binding of [3H]naloxone to small regions of adult rat brain. Reliable determinations of ligand binding were quantified with about 50 micrograms of protein per assay tube. Large differences in [3H]naloxone binding were obtained between various brain nuclei, and these differences were consistent with prior determinations of opiate receptor densities in various rat brain nuclei using autoradiographic techniques. PMID- 6292369 TI - Persisting nutritional neuropathy amongst former war prisoners. AB - Of 898 former Far East prisoners of war, assessed between 1968 and 1981, 49 (5.5%) had evidence of persisting symptomatic neurological disease dating back to their periods of malnutrition in captivity. The commonest syndromes were peripheral neuropathy (often of "burning foot" type), optic atrophy, and sensori neural deafness. Though nutritional neuropathies disappeared soon after release in most ex-Far East prisoners of war, in some they have persisted up to 36 years since exposure to the nutritional insult. PMID- 6292370 TI - Chronic idiopathic polyneuropathy treated with azathioprine. AB - The results of azathioprine therapy in five patients with chronic progressive or relapsing idiopathic inflammatory polyneuropathy are described. In four patients a sustained improvement followed treatment and in the other patient azathioprine successfully replaced corticosteroid therapy. The improvement was often delayed for up to three months. The literature on the use of azathioprine in chronic polyneuropathy is reviewed. We suggest that there is a place for azathioprine treatment in patients with chronic idiopathic polyneuropathy resistant or intolerant to corticosteroid treatment. PMID- 6292371 TI - Cranial computed tomography in the diagnosis of multiple sclerosis. AB - A group of 202 patients with suspected, probable or definite multiple sclerosis was studied, using cranial computed tomography (CT). Atrophy alone, or in combination with white-matter and periventricular lucencies, and areas of contrast enhancement, were the main abnormal findings in 52% of patients. Atrophy was detected in 44% of patients, and its frequency and severity correlated with disease duration up to 10 years, age, and disease category. Atrophic changes in the brainstem and cerebellum could be correlated with clinical data more often than supratentorial atrophy could be correlated with features such as dementia or mood changes. Lucencies in the white matter, thought to represent areas of demyelination, were noted in 21% of patients, and only a proportion of these lesions could be correlated with clinical data, the others being clinically silent. Contrast enhancement was seen in a small proportion of white-matter lesions, and was independent of disease activity and steroid medication. Electrophysiological tests and cerebrospinal fluid analysis showed a higher yield of abnormality than CT scanning in cases with suspected or possible multiple sclerosis, though in such patients CT scanning excluded alternative cerebral atrophy. Modifications of the technique of CT scanning may improve the detection rate of white-matter lesions, thereby enhancing the value of CT as a diagnostic tool in the study of patients with multiple sclerosis. PMID- 6292372 TI - Effects of cyclic AMP on S-100 protein level in C-6 glioma cells. AB - Dibutyryl cyclic AMP (dbcAMP) markedly elevated S-100 protein level in C-6 glioma cells in vitro. Quantitative analysis by two-dimensional polyacrylamide gel electrophoresis revealed that the elevation was caused by a combination of increased synthesis and reduced degradation of S-100 protein in C-6 cells exposed to dbcAMP. These results suggest that dbcAMP affects both the synthesis and the degradation of S-100 protein in C-6 cells. PMID- 6292373 TI - Branching pattern and properties of vertical- and horizontal-related excitatory vestibuloocular neurons in the cat. AB - 1. The axonal trajectories of excitatory vestibuloocular neurons and their synaptic contacts with extraocular motoneurons were studied by means of spike triggered signal averaging and microstimulation techniques. A majority of the excitatory neurons related to the vertical semicircular canals were located in the border of the descending and medial nuclei and the rostral half of the descending nucleus. 2. Individual vestibuloocular neurons activated by stimulation of the ampullary nerve of the anterior semicircular canal excited motoneurons within both the contralateral inferior oblique and contralateral superior rectus motoneuron pools. 3. Individual vestibuloocular neurons receiving input from the ampullary nerve of the posterior semicircular canal excited motoneurons in both the contralateral trochlear nucleus and contralateral inferior rectus motoneuron pools. The branching pattern of single vestibuloocular neurons activated by the anterior and posterior canals probably underlies conjugate eye movement during vertical head rotation. 4. Time to peak and shape indices of unitary excitatory postsynaptic potentials (EPSPs) suggested that the location of the synaptic contact of vestibuloocular neurons was on the soma or proximal dendrites of the target extraocular motoneurons. 5. In contrast, we did not find conclusive evidence that single vestibuloocular neurons receiving input from the horizontal semicircular canal give off axon collaterals to motoneurons innervating both the contralateral lateral rectus and the ipsilateral medial rectus muscles. Projection of horizontal vestibuloocular neurons to motoneurons supplying individual muscles might be useful for convergence during horizontal head movement. PMID- 6292374 TI - Specificity of electrical coupling among neurons innervating forelimb muscles of the adult bullfrog. AB - 1. The specificity of electrical connections among sensory fibers and motoneurons in the bullfrog's spinal cord was studied by recording intracellularly from brachial motoneurons. Synaptic potentials evoked by stimulation of individual muscle nerves were recorded in normal and reduced-calcium solutions and after acute section of dorsal or ventral roots. 2. Homonymous motoneurons are electrically coupled. After the dorsal roots were cut to abolish sensory input, short-latency potentials were almost always evoked anti-dromically in a motoneuron by stimulation of its own muscle nerve but rarely by stimulation of nerves innervating other muscles. These potentials differed from chemically mediated synaptic potentials in this preparation; they had a shorter latency and remained after perfusion with reduced-calcium solutions. This evidence suggests that they are mediated electrically. 3. Some motoneurons that innervate functionally equivalent muscles are electrically coupled. Approximately two thirds of the motoneurons innervating the internal or external heads of the triceps muscles received coupling potentials on stimulation of the other nerve, although never on stimulation of the heteronymous, medial, triceps nerve. 4. The monosynaptic potentials evoked by muscle sensory afferents in motoneurons often have both electrical and chemical components. The electrical component occurred with short delay and persisted in reduced-calcium solutions. The chemical component occurred 1.5-2.0 ms later, at 14 degrees C, and was abolished by reducing calcium in the bathing solution. Muscle sensory afferents make these mixed synapses on homonymous, heteronymous, and other motoneurons. PMID- 6292375 TI - Effects of temperature on identified central neurons that control jumping in the grasshopper. AB - Grasshoppers, like many poikilotherms, are generally more active at warmer body temperatures. In particular, they jump more frequently when warm. To determine the neuronal basis of this increase in jumping activity, we investigated the effects of temperature on the properties of identified central neurons known to be involved in the control of the jump; these included the fast extensor tibiae (FETi) motoneuron and the C, G, and M interneurons. Heating did not result in a reduction in the current or voltage threshold for action potentials; in most cases, there was an increase in the current threshold with heating. At higher temperatures, the frequency-current relations of interneurons and motoneurons had steeper slopes. With strong current pulses, increasing the temperature resulted in an increase in the initial peak firing frequencies of central neurons and usually also in their steady state firing frequencies. A second temperature effect favoring increased CNS activity in warm grasshoppers was increased afferent input from the periphery. In a broad variety of sensory receptors, there was a dramatic increase in their sensitivity to sensory stimuli at both threshold and suprathreshold intensities. Various identified central neurons differed in the way in which some of their properties were influenced by temperature. The C and G interneurons showed a striking similarity in the unusual way in which their repetitive firing properties were influenced by heating. Since these neurons are sibling progeny of a single neuroblast, this shared physiological property is correlated with their developmental history. PMID- 6292376 TI - Chronic effects of a monoamine oxidase-inhibiting antidepressant: decreases in functional alpha-adrenergic autoreceptors precede the decrease in norepinephrine stimulated cyclic adenosine 3': 5'-monophosphate systems in rat brain. AB - Various antidepressant drugs (monoamine oxidase inhibitors and tricyclics) enhance norepinephrine availability and lead to adaptive changes in brain noradrenergic systems, namely, decreases in the number of beta receptors and in the responsiveness of adenylate cyclase to norepinephrine monamine oxidase inhibitor, but not after 3 days, there is an increase in norepinephrine release from rat brain microsacs in response to 43 mM KCl stimulation. Microsacs prepared from 21-day clorgyline-treated animals also show a marked decrease in the inhibition of norepinephrine release caused by the alpha 2-selective agonist clonidine. These functional changes in norepinephrine release mechanisms are accompanied by a 53% reduction in brainstem alpha 2 receptor density as measured by [3H]clonidine binding. At the same time, despite findings of a decrease in beta receptor number as determined by [3H]dihydroalprenolol binding data, no significant decrease in the responses of cyclic adenosine 3': 5'-monophosphate (cyclic AMP) systems to norepinephrine stimulation is observed. Decreases in the cyclic AMP response are observed by day 35 of clorgyline treatment. The results provide direct physiological support for a change in the norepinephrine release mechanism and an effect on autoreceptors, specifically, preceding postsynaptic adaptive changes in the instance of one antidepressant, clorgyline. Difficulties in observing such changes with other antidepressants may result from the multiple nature of alpha-adrenergic receptors, especially as measured by radioactive ligand techniques; the lack of a direct relationship between physiological changes and receptors as measured by radioligand techniques; the large doses of monoamine oxidase inhibitors used in some studies; and the possible multiplicity of antidepressant molecular mechanisms. PMID- 6292377 TI - Electrophysiology of isolated hippocampal pyramidal dendrites. AB - In order to study the electrical properties of dendritic membranes independent of the effects of somatic potentials, intracellular recordings in guinea pig hippocampal slices were obtained from the dendrites of CA1 pyramidal neurons (HPCs) which had been isolated from their somata by cuts made through the proximal stratum radiatum. Spikes and subthreshold membrane responses to intracellular current pulses were compared in intact and isolated dendrites and in the residual portions of neurons whose apical dendrites had been severed ("isolated somata"). Isolated dendrites generated both fast, QX-314-sensitive, sodium-mediated spikes, and slow higher threshold spikes which were QX-314 resistant and presumably mediated by Ca2+. Depolarization of "isolated somata" ordinarily evoked only fast (Na+) spikes, but presumed Ca2+ spikes could be elicited after exposure to QX-314 (a local anesthetic). Anomalous inward rectification was depressed by QX-314 in somata but not in dendrites, suggesting that the ionic basis for subthreshold as well as regenerative conductances was different at different sites on the neuron. The dendritic membrane in CA2 HPCs thus generates both Na+- and CA2+-mediated spike potentials and a subthreshold response which probably is mediated primarily by CA2+. Attempts to describe the integrative functions of these neurons must take into account the variety of conductances which are activated nonuniformly in somata and dendrites by changes in membrane potential. PMID- 6292378 TI - Prenatal development of gabaergic, glycinergic, and dopaminergic neurons in the rabbit retina. AB - The main findings from our studies on the emergence and maturation of neurotransmitter systems in the rabbit retina are that, during normal development: (1) the commitments for certain neurons to become GABAergic, glycinergic, or dopaminergic are made prenatally; the maturation of these neurons, however, occurs postnatally; (2) specific accumulations of exogenously supplied transmitters by presumed retinal neurons were first observed autoradiographically around embryonic day 22 (E22) for gamma-aminobutyric acid (GABA), E25 for glycine, and E27 for dopamine, suggesting that putative GABAergic, glycinergic, and dopaminergic neurons are determined phenotypically at least by these days; (3) based upon the three transmitter-specific properties- uptake, synthesis, and release--the uptake property is generally the first to emerge, while the appearance of the other properties follows a precise and distinct temporal pattern for each of the transmitters studied; (4) for GABAergic and glycinergic systems, the emergence of K+-stimulated, Ca2+-dependent release mechanisms occurs many days after the first appearance of the uptake properties; in contrast, the mechanisms for dopamine uptake and release emerge at about the same time; and (5) by our criteria, putative GABAergic and glycinergic amacrine cells are mature between postnatal day 10 (P10) and P12, around the time when the eyes first open and direction-selective ganglion cells can first be recorded; however, the dopaminergic neurons probably are not mature until about P24. PMID- 6292379 TI - Inhibitor of adenosine 3':5'-monophosphate-dependent protein kinase blocks presynaptic facilitation in Aplysia. AB - Sensitization of the gill withdrawal reflex results from presynaptic facilitation at the excitatory synapses made by sensory neurons on gill motor neurons. Facilitation is accompanied by an increase in the duration of the action potential in sensory cells because of the depression of a K+ current. This results in an increasd influx of CA2+ and a greater release of transmitter from sensory neurons. There is evidence that serotonin is the facilitating transmitter and that the depression of the K+ current by serotonin mediated by cAMP-dependent protein phosphorylation. To test further the role of the cAMP-dependent protein kinase and of protein phosphorylation in sensitization, we have attempted to prevent or reverse the development of the electrophysiological correlates that accompany sensitization. We have pressure-injected sensory neurons with a specific and a stable protein inhibitor of the cAMP-dependent protein kinase both before and after the application of serotonin or the activation of the facilitator neurons. The increase in spike broadening that accompanies facilitation was prevented or diminished by injection of the inhibitor. Moreover, injection of the inhibitor could reverse fully the developed spike broadening produced by prior application of serotonin. These observations strenthen the evidence for the involvement of protein phosphorylation in presynaptic facilitation. Phosphorylation of the substrate protein evidently is quite labile and does not persist after the kinase is inhibited. Thus, the time course of short term sensitization appears to be determined by an active kinase. We think that it is likely that the mechanism for maintaining the kinase in an active form resides in the slow decay of the cAMP produced by the action of serotonin or the facilitator neurons on the sensory cells. PMID- 6292380 TI - Facilitatory transmitter causes a selective and prolonged increase in adenosine 3':5'-monophosphate in sensory neurons mediating the gill and siphon withdrawal reflex in Aplysia. AB - Sensitization of the gill and siphon withdrawal reflex in the marine mollusc, Aplysia california, is a simple form of learning Underlying this behavioral changes is a cascade of biochemical events. The first step in this cascade is postulated to be an increase in cAMP within the sensory neurons of the abdominal ganglion. We have developed a labeling protocol with 32Pi which permits us to measure the synthesis of cAMP within a single sensory neurons. Application of serotonin for 5 min was found to triple the content of [32P]cAMP in sensory neurons. The response is specific to serotonin: dopamine, a transmitter that does not produce sensitization, did not increase cAMP. Physiological stimulation of facilitator neurons also resulted in a 3.5-fold increase of cAMP in sensory neurons but not in other cells of the ganglion. We studied the time course of the increase of cAMP in sensory cells stimulated with serotonin and found that it parallels closely the time course of the short term form of presynaptic facilitation. We also have determined the effects of transmitters on the synthesis of cAMP in other identified neurons of the ganglion. The bag cells responded specifically to serotonin. R15, which has been shown to be hyperpolarized both the serotonin and by dopamine, responded to both transmitters by increased synthesis synthesis of cAMP. Thus, the dopamine- and serotonin sensitive cyclase can be localized to both the same and different cells. Other cells did not respond to serotonin or to dopamine, indicating that a transmitter sensitive adenylate cyclase is a specific property and is not present in all neurons. PMID- 6292381 TI - Chloride-dependent enhancement by barbiturates of gamma-aminobutyric acid receptor binding. PMID- 6292382 TI - Vascularization of the ear: normal-variations-glomus tumors. PMID- 6292383 TI - Isomorphus ionic replacement: experimental evidence for the hypothesis proposed to explain gallium-67 accumulation. PMID- 6292384 TI - Influence of cardiac factors on the regional arm-brain mean transit time. PMID- 6292385 TI - Absence of respiratory effects in subjects exposed to low concentrations of TDI and MDI. AB - One hundred seven subjects from a polyurethane plastic manufacturing plant have been followed over a five-year period with measurements of forced expiratory volume in 1 s (FEV1), and questionnaires on respiratory symptoms and smoking habits. Environmental concentrations of toluene diisocyanate and diphenyl methyl diisocyanate were extensively monitored to provide accurate estimates of the upper-limits of exposure of the subjects. Current mean levels of FEV1 in this population were higher than those predicted for healthy subjects. The five-year change in FEV1 did not exceed that expected from aging. No acute change in FEV1 could be demonstrated over the course of a Monday either before or after a two week vacation. No improvement in ventilatory function was observed over the vacation period. The presence of cough or sputum was related to smoking but was not related to isocyanate exposure. The results indicate that exposure of workers to extremely low levels of isocyanates (time-weighted average concentrations of the order of 0.001 parts per million [ppm]) is not associated with chronic respiratory symptoms or effects on ventilatory capacity. The results suggest that isocyanates can be controlled to the point of eliminating effects as measured by these techniques. PMID- 6292386 TI - Addictive processes. PMID- 6292388 TI - The effect of sodium bicarbonate and hydrogen peroxide on the microbial flora of periodontal pockets. A preliminary report. AB - This study investigated the effects of sodium bicarbonate and hydrogen peroxide on the microbial flora of 4 to 7 mm periodontal pockets. Four selected patients were instructed to brush assigned quadrants daily with a sodium bicarbonate-3% hydrogen peroxide paste utilizing a split-mouth design. The paste was pumped between the teeth and gingiva via sulcular brushing and a Perio Aid. The control sides were treated in the identical manner with fluoridated paste. Probing depths on each patient were taken on Days 1 and 21 and examined under dark-field microscopy. For each sample, morphology and motility of all cells were recorded. The data were analyzed by three factor repeated measures analysis of variance. No statistically significant differences were found between the control and medicament sites at the 0.005 level regarding changes in the microbial flora. Probing depths were reduced during the study. However, similar results were found on both medicament and control sites. It appeared that sodium bicarbonate and hydrogen peroxide have no unusual benefits in reducing the microbial flora of periodontal pockets. PMID- 6292387 TI - Prevalence and importance of congenital cytomegalovirus infection in three different populations. AB - A Chilean population was compared to low-income and middle/upper-class populations in Birmingham, Ala., with regard to prevalence of congenital cytomegalovirus infection as well as the importance of this infection in neonatal deaths. In the highly seroimmune Chilean (98%) and low-income Birmingham (82%) groups, congenital infections occurred more often (1.7% and 1.9%, respectively) than in the less immune (56%) middle/upper-income group in Birmingham (0.6%). In 407 autopsies reviewed in Chile no neonatal deaths were attributed to cytomegalic inclusion disease, whereas in Birmingham cytomegalovirus was the cause of death in nine of 938 (1%) newborn infants. These findings further support the concept that, despite an apparent lack of protection against intrauterine transmission, maternal immunity reduces the risk of severe fetal infection. PMID- 6292389 TI - Determination of sulfadiazine and N4-acetylsulfadiazine in biological fluids by liquid chromatography on silica gel with an aqueous buffer as mobile phase. AB - Sulfadiazine and N4-acetylsulfadiazine were determined in biological fluids by the direct injection (plasma after protein precipitation and urine after dilution 100 times) of 20 microliters on a silica gel column. The mobile phase was an aqueous citrate buffer (pH 4.0) and UV detection was a 264 nm. Chromatographic selectivity was optimized by the silica gel surface and pH of the mobile phase. Detection limits were approximately 0.4 microgram/ml for sulfadiazine in plasma and approximately 5 and 7 micrograms/ml of sulfadiazine and N4-acetylsulfadiazine in urine, respectively. In quantitations by peak heights relative to an internal standard (sulfamerazine), within-run precisions (srel%) for sulfadiazine were 1.7 and 4.0% at 40 and 2 micrograms/ml, respectively, in plasma and 0.76 and 1.7% and 25 micrograms/ml, respectively, in urine. PMID- 6292390 TI - Determination of bacmecillinam, an amdinocillin prodrug, in human and canine whole blood by reversed-phase liquid chromatography. AB - Bacmecillinam is an amdinocillin prodrug designed to be easily hydrolyzed in biological materials, so special procedures were developed for the collection of blood specimens. Whole blood was collected in tubes containing bacampicillin as an adsorption inhibitor and kept at -70 degrees; the extracting solvent, hexane methylene chloride (9:1, v/v), was added to the cold tubes, and the extraction was performed during the thawing of the samples. The organic phase was partially evaporated before a reextraction to a small volume of acidic aqueous phase was made. The separation was performed on a microparticulate. C18-alkyl bonded silica packed in glass-lined stainless steel columns. Mobile phase was a buffer (pH 6) acetonitrile mixture containing N-hexyl-N-methylamine as an adsorption inhibitor. The detection limit was 600 pg/ml of whole blood, and the within-run precision (srel%) was approximately 8% at the 5-ng/ml level. PMID- 6292391 TI - Biological maturation and beta-adrenergic effectors: pre- and postnatal development of the adenylate cyclase system in the rabbit heart. AB - The relationship between biological maturation and adenylate cyclase activity was studied in membrane preparations of rabbit ventricular muscle. Basal adenylate cyclase activity was lower in the adult than in the 1-day-old neonate or 27-day old fetus. Maximal stimulation of adenylate cyclase by isoproterenol was 2.5 times greater and the EC50 values were 2-fold higher in the adult than the 27-day old fetus or 1-, 7- and 12-day-old neonate. No significant differences in isoproterenol- or Mg++-stimulated activity were observed among the younger age groups nor was the Mg++-stimulated Vmax of adenylate cyclase significantly affected by biological maturation. Sodium fluoride, guanyl-5'yl imidodiphosphate and GTP also stimulated adenylate cyclase activity in a dose-dependent fashion similar to isoproterenol. Sodium fluoride (2.5-10 mM) increased adenylate cyclase activity in the adult to a significantly greater extent than the 1-day-old neonate. Guanyl-5'yl imidodiphosphate and GTP (0.1-10.0 microM) augmented adenylate cyclase activity to approximately the same degree (although some small differences were observed) in the fetus, neonate and adult. However, when guanyl 5'yl imidodiphosphate was preincubated with membrane preparations before in vitro assay, adenylate cyclase activity was increased 10-fold in the adult, whereas membranes from 1-day-old animals were unaffected. These data suggest that the processes regulating hormonal and pharmacological activation of adenylate cyclase are modified during biological maturation. PMID- 6292392 TI - Characterization of [3H]yohimbine binding to putative alpha-2 adrenergic receptors in neonatal rat lung. AB - In contrast to the adult rat lung, which lacks alpha-2 adrenergic receptor binding sites, the neonatal rat lung had a high density of [3H]yohimbine binding sites. The binding of this alpha-2 antagonist was saturable, reversible and stereospecific. Steady-state binding was reached by 15 min at 23 degrees C, and the association and dissociation rate constants were 0.17 min-1nM-1 and 0.30 min 1, respectively. From saturation experiments, a Bmax of 304 fmol/mg of protein and a KD of 1.53 nM were calculated. Surprisingly, no appreciable specific binding was observed in this tissue for the alpha-2 agonists [3H]clonidine, [3H]paraminoclonidine and [3H]epinephrine. Inhibition experiments indicated that the binding site had the characteristics of an alpha-2 adrenergic receptor although yohimbine was only 5 times more potent than prazosin. The Ki for epinephrine was decreased 4-fold by 100 microM guanyl-5'-6'-imidodiphosphate suggesting that the receptor binding site may be coupled to adenylate cyclase. Ethanol inhibited the [3H]yohimbine binding with an IC50 of 200 mM. This inhibition was "competitive-like" as increasing concentrations of ethanol resulted in an increased KD, but not a decrease in Bmax. The presence of high density [3H]yohimbine binding sites in early life may have a developmental importance which is not yet understood. PMID- 6292393 TI - Discriminative stimuli produced by clonidine: an investigation of the possible relationship to adrenoceptor stimulation and hypotension. AB - In a leverpressing operant procedure, male rats were trained to respond for food reinforcement on one lever after an injection of clonidine (0.04 mg/kg) and to respond on an alternate lever for food reinforcement after an injection of saline. All 36 rats learned to discriminate the drug reliably from saline, thereby indicating that clonidine produces discriminative interoceptive stimuli. The discriminative stimulus was both dose- and time-dependent, with an ED50 of 0.018 mg/kg and an optimum time of action occurring from 15 to 60 min after injection. Although clonidine produced a reduction in response rate, this was not the basis of the discriminative stimulus as other drugs with similar depressant action did not generalize. The clonidine stimulus was dose-dependently antagonized by the alpha-2 adrenergic antagonist, yohimbine, whereas receptor antagonists of alpha-1 adrenergic, beta adrenergic, dopaminergic, serotonergic, cholinergic or opioid systems were ineffective in blocking the interoceptive stimulus produced by clonidine Lofexidine, guanabenz and methyldopa, all centrally acting hypotensive drugs that act through alpha-2 adrenoceptor mechanisms dose-dependently generalized to the clonidine cue, whereas hydralazine, minoxidil, propranolol and prazosin, hypotensive drugs acting through other mechanisms, did not generalize. These results suggest that clonidine produces interoceptive stimuli that are discriminable by rats and mediated through central alpha-2 adrenoceptor stimulation. PMID- 6292394 TI - Blockade of forebrain gamma-aminobutyric acid (GABA) receptors and reflex activation of the cardiac vagus in anesthetized cats. AB - gamma-Aminobutyric acid (GABA) antagonists have been shown to produce sympathetically mediated increases in blood pressure and heart rate when restricted to the forebrain cerebral ventricles of anesthetized cats. This study explored the possibility that similar administration of these agents might produce reciprocal effects on reflex cardiac vagal excitability. Drugs were infused into and restricted to the forebrain ventricles of cats anesthetized with chloralose and urethane. Arterial pressure and heart rate were continuously monitored and reproducible reflex vagal bradycardia was periodically elicited by bolus i.v. injections of phenylephrine. In early experiments in intact cats and in later studies in spinal transected animals, i.c.v. administration of the GABA antagonist bicuculline methiodide (1-32 micrograms) suppressed phenylephrine induced reflex bradycardia in a dose-related fashion. When tested in spinal transected cats, i.c.v. picrotoxin, another GABA antagonist, mimicked this effect of bicuculline methiodide. Intraventricular muscimol (10 micrograms), a GABA agonist, had no effect in untreated cats but reversed the effects of bicuculline methiodide and picrotoxin. These data point to tonic GABAergic inhibition in the periventricular forebrain which suppresses the activity of a descending vagal inhibitory mechanism. PMID- 6292395 TI - Maturation of sympathetic neurotransmission in the rat heart. IX. Development of transsynaptic regulation of cardiac adrenergic sensitivity. AB - The number of beta adrenoceptors and the cardiac sensitivity to adrenergic stimulation increase substantially in the immediate postnatal period of the rat. To determine whether transsynaptic input influences this developmental process, the effects of a sympathomimetic and of agents which destroy noradrenergic nerve terminals on regulation of adrenergic postsynaptic sensitivity were compared in hearts from adult and developing rats. In mature animals, chronic exposure to the beta agonist isoproterenol (2.5 mg/kg s.c.) led to rapid onset (3-5 days) of chronotropic adrenergic subsensitivity accompanied by a loss of beta adrenoceptor binding sites; chemical sympathectomy by daily administration of guanethidine (50 mg/kg s.c.) or by 6-hydroxydopamine (100 mg/kg s.c. given once daily for 3 days) resulted in chronotropic adrenergic supersensitivity and increases in binding sites. These data in the adult agree with classical transsynaptic modulation of adrenergic postsynaptic reactivity. In contrast, identical drug treatments of immature rats beginning 1 day after birth failed to evoke changes in either chronotropic adrenergic sensitivity or in numbers of beta adrenoceptor binding sites until the 3rd to 4th week. Consequently, the initial development of beta adrenoceptors and responsiveness to catecholamines in the neonatal myocardium are not transsynaptically regulated; rather, other (e.g. hormonal) factors appear to control early maturation of cardiac adrenergic sensitivity. PMID- 6292396 TI - Accumulation of hypothalamic endorphins after repeated injections of anorectics which release serotonin. AB - Two anorectic drugs which stimulate serotonin neuronal activity by releasing serotonin from nerve terminals, d-fenfluramine and CM 57 277 (4-amino-[6-chloro-2 pyridyl]-1-piperidine HCl), were studied in rats. Both drugs, when given for 5 days at doses of 15 mg of d-fenfluramine per kg/day and 20 mg of CM 57 277 per kg/day, decreased body weight gain and increased content of Met5-enkephalin and beta-endorphin in hypothalamus, but not in frontal cortex or pituitary. The increase in the hypothalamic content of the two opioid peptides elicited by d fenfluramine was reversed by metergoline and p-chlorophenylalanine, suggesting that it is mediated by serotonin. The content of two other hypothalamic neuropeptides, cholecystokinin and substance P, were not affected by d fenfluramine. Although Met5-enkephalin content in striatum was increased transiently by d-fenfluramine, this was not a serotonin-mediated effect because it was not abolished by metergoline. The decrease in body weight gain was prevented by metergoline, but not by i.p. injection of p-chlorophenylalanine. The effect of p-chlorophenylalanine on intestine may contribute to its failure in reversing the anorectic effect of d-fenfluramine. Naltrexone, an antagonist of opiate receptor, decreased body weight gain but exerted no effect on hypothalamic Met5-enkephalin or beta-endorphin content. Taking into consideration that the increase in Met5-enkephalin and beta-endorphin may have resulted from accumulation due to decreased utilization, the anorectic effects of serotonin releasing drugs may be mediated by a reduction in the functional role of hypothalamic opioid peptides. PMID- 6292397 TI - A comparison of the pharmacological properties of Clostridium botulinum type C1 and C2 toxins. PMID- 6292398 TI - Enhancement of prostaglandin output during activation of beta-1 adrenoceptors in the isolated rabbit heart. AB - The purpose of this study was to elucidate the type of adrenoceptor that mediates the effect of adrenergic stimuli on prostaglandin (PG) synthesis in the isolated rabbit heart and to determine the relationship of the released PGs to the mechanical changes elicited by catecholamines and stimulation of the cardiac sympathetic nerves. The output of 6-keto PGF1 alpha, PGE2 and PGF2 alpha was increased by electrical stimulation of the sympathetic nerves, norepinephrine, isoproterenol, dobutamine and angiotensin II, but not by phenylephrine or isoetharine. Propranolol or atenolol, but not phentolamine or butoxamine, blocked the output of PGs elicited by adrenergic stimuli. Indomethacin prevented the increase in PG formation caused by all stimuli. Moreover, the adrenergically induced release of PGs was not related to changes in heart rate, systolic tension or vascular tone elicited by the adrenergic stimuli. These data indicate that the adrenergically induced release of PGs in the isolated rabbit heart is due to the activation of beta-1 adrenoceptors and is independent of the mechanical effects produced by the adrenergic stimuli. PMID- 6292400 TI - Alterations in prejunctional responsiveness to angiotensin II during the development of 2-kidney, 1-clip Goldblatt hypertension in rats. PMID- 6292399 TI - GABAergic modulation of ethanol-induced motor impairment. AB - Direct or indirect pharmacological manipulation of gamma-aminobutyric acid (GABA) receptor activity was examined in relation to the motor incoordinating actions of ethanol in the rat. Ethanol (1.13-3.0 g/kg i.p.) caused a dose-dependent increase in the height of aerial righting. This motor impairment was increased selectively by intracisternal injection of the GABA agonists muscimol (0.10 microgram), 4,5,6,7-tetrahydroisoxazole(5,4-c) pyridin(3-ol) (1.0 microgram) and GABA (1000 micrograms). The GABA antagonist, bicuculline (1.0 and 5.0 micrograms intracisternally), reduced impairment. Thus, direct manipulation of GABA receptor activity modulated motor incoordination caused by ethanol. In addition, indirect acting GABA-mimetics, such as gamma-acetylenic GABA (100 mg/kg i.p.), aminooxyacetic acid (50 mg/kg i.p.), ethanolamine-O-sulfate (250 mg/kg i.p.) and L-2,4-diaminobutyric acid (600 mg/kg i.p.) all potentiated the increase in the height of aerial righting caused by ethanol treatment. Failure of ethanol to modify the binding of [3H]muscimol to cerebral cortical membranes in vitro suggested there was no direct competition for GABA binding sites or facilitation of the binding of GABA to these sites by ethanol. Also, no simple relationship was observed between the degree of motor impairment caused by either ethanol or gamma-acetylenic GABA and changes in GABA concentration in three brain areas. Although GABAergic neurons may be involved in the mechanism underlying ethanol induced depression of motor coordination, the interaction does not involve a direct activation of GABA receptors by ethanol. PMID- 6292401 TI - Separate noradrenergic receptors could mediate clonidine-induced antinociception. AB - The antinociceptive effect of clonidine on a response to painful stimulation mediated by supraspinal structures was recorded after s.c. administration of the drug in doses from 50 up to 2000 micrograms/kg. Both low and high doses of clonidine produced antinociception on the pain threshold studied. A careful analysis of the dose-response curve showed, however, that the net effect recorded involved the sum of responses from at least two functional systems or receptor sites. When the dose-response relationship of clonidine-induced antinociception was studied after alpha-1 receptor blockade by means of phenoxybenzamine, it was found that this effect comprised contributions from different neurotransmitter systems. These results are discussed in terms of the possibility that separate adrenergic receptors mediate clonidine antinociception at different levels in the pain transmission. The determinant of the population of receptors being activated after systemic administration of clonidine is the dose given. PMID- 6292402 TI - Influence of repeated administration of desmethylimipramine on beta adrenergic and muscarinic cholinergic receptors and 45Ca++ binding to sarcoplasmic reticulum in the rat heart. PMID- 6292403 TI - Effect of ethanol-containing liquid diet upon gonadotropin receptor depletion in rat testis. AB - By using pair-feeding technique, the number of gonadotropin binding sites per unit mass of testicular homogenate was measured 15, 30, 45 and 60 days after ethanol administration to 60-day-old male rats. The ingestion of ethanol (5%, v/v) as a part of a nutritionally adequate liquid diet resulted in 30, 35 and 40% reduction in gonadotropin receptors by 15, 30 and 45 days, respectively. The body weights in the alcohol-fed rats were comparable to pair-fed controls. These findings suggest that the presence of ethanol in the liquid diet depresses the level of gonadotropin receptors in the rat testis. PMID- 6292404 TI - Sodium currents and sodium-current fluctuations in rat myelinated nerve fibres. AB - 1. Sodium currents and fluctuations of sodium currents were measured in myelinated fibres of rat sciatic nerve under voltage clamp at 20 degrees C.2. Relaxations of sodium currents during various test potentials were recorded in the presence of 6 nM-TTX in the extracellular solution. The activation of sodium currents at low depolarizations could be described with the m(2) formulation. At increasing potentials higher powers of m up to 4 were required. The mid-point of the P(Na) (E) curve was located near E = -32 mV. Sodium inactivation during various depolarizations developed in two phases.3. The resistance in series with the nodal membrane was calculated from peak sodium currents without and with 6 nM TTX in the extracellular solution. The resistance varied between different fibres and ranged between 190 and 620 kOmega.4. From peak sodium currents at the same mambrane potential without and in the presence of TTX an apparent equilibrium dissociation constant of 1.6 nM was calculated for TTX binding to sodium channels.5. The conductance gamma and the number N(0) (corrected for series resistance effects) of sodium channels were evaluated from ensemble average values of the mean sodium current and the variance of sodium-current fluctuations at the beginning of a test pulse. The mean values were gamma = 14.5 pS, N(0) = 21,000 per node.6. The spectral density of stationary sodium-current fluctuations exhibited two relaxation components whose time constants were comparable to those of sodium activation and inactivation. At low depolarizations the variance produced by inactivation fluctuations was larger than predicted by the m(3). h formulation.7. It is concluded that individual sodium channels of rat and frog nerve have similar gating properties. In mammalian nodes the number of sodium channels is lower and the single-channel conductance higher than in amphibian nodes. PMID- 6292405 TI - Enhancement of ionic currents through voltage-gated channels in the mouse oocyte after fertilization. AB - 1. The changes of voltage-gated ion channels in the mouse oocyte after fertilization were investigated under voltage clamp.2. About 60 min after introduction of sperm suspension into the fertilization medium, the amplitude of inward current through Ca(2+)-channels increased, which occurred at anaphase during the second meiotic division. The peak amplitude of the maximum inward current per unit membrane capacity of the oocytes at metaphase was 20+/-3 muA/muF in 50 mM-Sr medium. It was 28+/-8 muA/muF at anaphase, and 32+/-5 muA/muF at telophase. The kinetic properties as well as selectivity among Ca, Sr and Mn ions were not altered after fertilization.3. The outward surge current which was found at the higher membrane potential over +50 mV also increased in amplitude after fertilization, simultaneously with the increase in amplitude of inward current through Ca(2+)-channels. The means and the standard deviations of the surge current per unit membrane capacity at 120 mV were 31+/-8 muA/muF at metaphase, and 48+/-7 muA/muF at telophase. The kinetic properties of the outward surge current were not altered after fertilization.4. Application of colcemid (10(-7) mole/l.) or cytochalasin B (2 x 10(-5) mole/l.) did not prevent the increase in amplitude of both inward current through Ca channels and the outward surge current.5. The membrane currents in N-18 mouse neuroblastoma cells in logarithmic growth phase were examined under voltage clamp. The N-18 neuroblastoma cells possessed the Ca inward current and the delayed outward current. The kinetic properties and the steady-state inactivation of Ca(2+)-channels in N-18 neuroblastoma cells were compared with those in mouse oocytes. It was concluded that they could be regarded as identical between the mouse oocyte and the N-18 neuroblastoma cell. PMID- 6292406 TI - The role of spinal cord transmission in the ventilatory response to electrically induced exercise in the anaesthetized dog. AB - 1. The ventilatory response to electrically induced ;exercise' was studied in six chloralose-anaesthetized dogs. The on-transient and steady-state responses to ;exercise' were compared in the same dogs before and after spinal cord transection at T8/9 (dermatome level T6/7) on fifteen occasions.2. Phasic hind limb ;exercise' was induced for periods of 4 min by passing current (2 Hz modulated 50 Hz sine wave) between two needles inserted through the hamstring muscles. The maximum current used was 30 mA. This was below the level previously found to produce an artifactual stimulation of breathing with the cord intact.3. Cord transection produced no significant change in either the resting values of ventilation ( V(I)) and CO(2) production ( V(CO) (2)) or the ventilatory equivalent for CO(2) during ;exercise' ( big up tri, open V(I)/ big up tri, open V(CO) (2)).4. During the steady state of exercise P(a, CO) (2) was on average significantly lower than at rest with the cord intact (mean big up tri, openP(a, CO) (2), - 2.1 mmHg; range - 5.7 to + 1), and higher, though not significantly, with the cord cut (mean P(a, CO) (2), + 1.2 mmHg; range - 1.5 to + 4.3). However, even in the absence of spinal cord transmission, the ventilatory response to exercise could not be accounted for on the basis of CO(2) sensitivity; the big up tri, open V(I)/ big up tri, openP(a,CO) (2) obtained with exercise (apparent sensitivity) was significantly greater than that obtained with CO(2) inhalation (true sensitivity) both before and after cord section.5. V(I) and V(CO) (2) increased more slowly with the cord cut than with the cord intact. This was thought to be due to a slower increase in venous return in the absence of sympathetic innervation of the lower half of the body following cord transection.6. Similar experiments were performed during muscle paralysis (following gallamine triethiodide). Ventilation was maintained with a respirator controlled by phrenic nerve activity. These experiments showed an increase in ventilation, independent of muscle contraction, which was only present when the cord was intact and which was confined to the on-transient. Only in the absence of spinal cord transmission could there be certainty that the dynamics of the ventilatory response to electrically induced ;exercise' was free of artifact.7. It was concluded that spinal cord transmission is not necessary for the steady state ventilatory response to electrically induced exercise of the hind limbs.8. The dog with spinal cord transection provides a suitable model for the study of the chemical control of breathing during electrically induced exercise. PMID- 6292407 TI - Block and activation of the pace-maker channel in calf purkinje fibres: effects of potassium, caesium and rubidium. AB - 1. The effects of low concentrations of Cs(+) (0.01-3mM) on the fully activated I V relation i(f)(E) for the pace-maker current in calf Purkinje fibres have been investigated. The action of Cs(+) is two-fold: in the negative region of the I-V curve Cs(+) induces a channel blockade; on the other hand, at more positive potentials Cs(+) can produce the opposite effect, i.e. a current increase.2. Cs(+)-induced blockade is concentration- and voltage-dependent, as observed on other cation channels. Data in the far negative voltage range (about - 150 to - 50 mV) can be fitted by a simple block model (Woodhull, 1973), which gives a mean value of 0.71 for the fraction of membrane thickness (delta) crossed by Cs(+) ions before reaching the blocking site. The value of delta does not appear to be affected by either external Na or external K concentrations. Values for the dissociation constant of the blocking reaction at E = 0 mV (k(0)) are found in the range 0.5-3.7 mM. In the positive region of the i(f)(E) relation the current depression caused by channel blockade vanishes. Unexpectedly, in this range the current can be observed to increase with Cs(+), and i(f)(E) curves in different Cs(+) concentrations show cross-over.3. Changing external K(+) also produces similar cross-over phenomena. Investigation of this effect reveals that the increase in slope of the I-V curve on raising the external K(+) concentration follows Michaelis-Menten kinetics, and can be interpteted in terms of K(+) induced channel activation. It is found that 44+/-6 mM-K(+) half-saturates the channel activating reaction.4. The Cs(+)-induced current increase is large in low K(+) solutions and vanishes in high-K(+) solutions, suggesting a competition between Cs(+) and K(+) ions in their activating action. Increasing Na(+) also limits the Cs(+)-induced current increase.5. Rb(+) also blocks the i(f) channel, though less efficiently than Cs(+). The block caused by Rb(+) is, unlike that of Cs(+), nearly voltage-independent, and is explained by assuming that the blocking reaction occurs near the external mouth of the channel (mean value of delta is 0.05). The zero-voltage dissociation constant (k(0)) of the Rb(+)-blocking reaction ranges between 1.4 and 5.4 mM, and is lower in low-Na(+), high-K(+) solutions.6. A possible characterization of the i(f) channel which explains these results includes an inner ;blocking' site, to which external Cs(+) ions bind, blocking the channel, and a more external ;activatory' site, to which K(+), Cs(+), Rb(+) and possibly Na(+) ions bind. Binding of K(+) to this site induces a current increase either by modulating the channel, or actually by opening the channel itself. A similar mechanism can apply to Cs(+) and to Rb(+) binding. PMID- 6292408 TI - Long-term potentiation of excitatory synaptic transmission in the rat hippocampus: the role of inhibitory processes. AB - 1. The possibility that changes in inhibitory processes are responsible for long term potentiation (l.t.p.) was examined using the rat hippocampal slice preparation.2. Inhibitory pathways were characterized using both extra- and intracellular recordings from the CA1 pyramidal cell layer. Stimulating electrodes were placed in either stratum radiatum or the alveus to allow orthodromic or antidromic activation of the pyramidal cells.3. Using extracellular recordings, inhibition was studied by applying paired pulses at interstimulus intervals of 20-500 msec through either the same or different stimulating electrodes, and quantifying the reduction in the population spike. An antidromic conditioning pulse was least effective in influencing the test response, while paired stimuli delivered through separate stimulators in stratum radiatum revealed the longest duration effects. Inhibition was either reduced or enhanced, depending upon the stimulation paradigm, with increasing stimulus intensity.4. With l.t.p., alterations in paired-pulse inhibition were observed corresponding to the changes in conditioning pulse amplitude. Reducing stimulus intensity to restore the initial conditioning pulse amplitude eliminated these effects.5. Using intracellular recordings, the effects of l.t.p. on inhibition were studied by examining changes in e.p.s.p.-i.p.s.p. sequences, i.p.s.p.s evoked by antidromic stimulation, and spontaneous depolarizing i.p.s.p.s observed with KCl-filled electrodes.6. Following l.t.p. enhanced e.p.s.p.s and slightly reduced, but prolonged, i.p.s.p.s were observed in response to orthodromic stimulation. Antidromically evoked, as well as spontaneous, i.p.s.p.s were unaffected.7 It is concluded that alterations in inhibitory processes are not responsible for l.t.p. in hippocampal subfield CA1. However, changes in the strength of inhibitory synapses as a consequence of long-term potentiation may modify the functional character of the hippocampal connexions. PMID- 6292409 TI - Effects of nystatin-mediated intracellular ion substitution on membrane currents in calf purkinje fibres. AB - 1. Calf cardiac Purkinje fibres were exposed briefly to the ionophore nystatin to promote exchange of caesium for intracellular potassium. The effects of Cs loading were stable for at least 30 min, but they could be reversed by nystatin mediated K loading.2. After Cs loading, the resting potential shifted to about 20 mV and the current-voltage relationship showed a strong inhibition of inwardly rectifying K channels.3. Anodal break stimulation evoked normal action potential upstrokes and twitch contractions. The early repolarization (phase 1) was markedly slowed.4. Cs loading simplified the pattern of current changes evoked by step depolarizations over the plateau range. Membrane current reached an inward peak and then declined monotonically.5. The current signal showed no hint of the transient outward current found in untreated or K-loaded preparations. Furthermore, Cs loading abolished the outward tails associated with deactivation of transient outward current, and occluded the blocking effect of the K-channel inhibitor 4-aminopyridine.6. Inhibition of transient outward current revealed a maximal inward current of about 5 muA/muF in 5.4 mM-Ca(o), which is considerably larger than the net inward current without Cs loading.7. The inward current was attributed to Ca channels on the basis of its sensitivity to membrane potential, extracellular Ca, D600, Mn and Cd.8. Cs loading also reduced slow current changes associated with delayed rectification and pace-maker depolarization.9. The results support the hypothesis that the transient outward current is carried by K(+) ions, while providing a method for unmasking inward Ca current. PMID- 6292411 TI - Prostaglandins and dermatology. The Ingram Lecture 1982. PMID- 6292410 TI - Enhancement of calcium current during digitalis inotropy in mammalian heart: positive feed-back regulation by intracellular calcium? AB - 1. Effects of digitalis compounds on slow inward Ca current I(si)) and contractile force were examined in ferret ventricular muscle (single sucrose-gap voltage clamp) and calf Purkinje fibres (two micro-electrode voltage clamp).2. In ventricular muscle, ouabain increased I(si) and inward current tails associated with I(si) conductance. The enhancement of I(si) followed a time course similar to the development of the positive inotropic effect, and it could be observed in the absence of aftercontractions or other signs of toxicit.3. The response of myocardial I(si) and twitch force to ouabain depended strongly on a previous history of driven action potentials.4. Veratridine, a toxin that promotes Na entry through tetrodotoxin-sensitive channels, also increased I(si) and twitch force in driven ventricular muscle preparations.5. The effects of ouabain, action potential stimulation and veratridine are consistent with reported effects of K poor solutions in indicating that elevation of intracellular Na can lead to enhancement of I(si). Additional experiments suggest that the link between Na(i) and I(si) involves intracellular Ca.6. When Cs-loaded Purkinje fibres were bathed in solutions containing Sr instead of Ca, enhancement of I(si) by strophanthidin was abolished even though a positive inotropic response persisted.7. After intracellular injection of Purkinje fibres with EGTA, I(si) no longer increased with strophanthidin, although it remained responsive to adrenaline.8. Clear-cut increases in I(si) were seen in Cs-loaded Purkinje fibres even at very low concentrations of strophanthidin (20-50 nM), where the occurence of Na pump inhibition has been questioned.9. Positive regulation of Ca entry by intracellular Ca may act as a facilitory mechanism that amplifies myocardial responsiveness to digitalis and other inotropic interventions. Through changes in I(si), small rises in diastolic free Ca might lead to large increases in the activator Ca transient during contraction. PMID- 6292412 TI - Bread for health and bread for slimming. PMID- 6292413 TI - Environmental and occupational hazards to the fetus. PMID- 6292414 TI - Newly recognized syndrome in the neck. PMID- 6292415 TI - Effects of acupuncture in bronchial asthma. PMID- 6292416 TI - Amoebiasis. PMID- 6292417 TI - Hydroxy- and amino-substituted piperidinecarboxylic acids as gamma-aminobutyric acid agonists and uptake inhibitors. AB - The syntheses of (3RS,4RS)-4-hydroxypiperidine-3-carboxylic acid (4), (3RS,5SR)-5 hydroxypiperidine-3-carboxylic acid (20), (3RS,4SR)-4-acetamidopiperidine-3 carboxylic acid (10), and (3RS,5SR)-5-acetamidopiperidine-3-carboxylic acid (18), related to the specific gamma-aminobutyric acid (GABA) uptake inhibitors (RS) piperidine-3-carboxylic acid (nipecotic acid) and (3RS,4SR)-4-hydroxypiperidine-3 carboxylic acid (21), are described. Furthermore, (3RS,4SR)-3-hydroxypiperidine-4 carboxylic acid (14), related to the specific GABA agonist piperidine-4 carboxylic acid (isonipecotic acid), has been synthesized. The structures of 4, 10, 14, 18, and 20 have been established by 270-MHz 1H NMR spectroscopic analyses. The affinity of the compounds for the GABA receptors and for the neuronal (synaptosomal) GABA uptake system in vitro has been measured. Compound 14 interacts selectively with the GABA receptors but less effectively than isonipecotic acid and the cis-isomer 22. Compounds 4, 18, and 20 are inhibitors of the GABA uptake system, although much weaker than nipecotic acid and (3RS,4SR) 4-hydroxypiperidine-3-carboxylic acid (21). Compound 10 is inactive in both test systems. PMID- 6292418 TI - Species- or isozyme-specific enzyme inhibitors. 8. Synthesis of disubstituted two substrate condensation products as inhibitors of rat adenylate kinases. PMID- 6292419 TI - Pentasubstituted quercetin analogues as selective inhibitors of particulate 3':5' cyclic-AMP phosphodiesterase from rat brain. AB - Some penta-O-substituted analogues of quercetin were synthesized and tested for the inhibition of cytosolic and particulate rat brain cyclic AMP and cyclic GMP phosphodiesterase activities. Ten of these compounds are potent and highly selective inhibitors of cAMP hydrolysis with respect to cGMP hydrolysis. They inhibit more potently the particulate enzyme than the cytosolic preparation. The highest selectivity was observed with penta-O-ethylquercetin and analogue 6d, which proved to be more selective and more potent inhibitors than the reference compound Ro 20-1724. Some structure-activity relationships are discussed. PMID- 6292420 TI - Synthesis and pharmacology of metabolically stable tert-butyl ethers of morphine and levorphanol. AB - 3-O-tert-Butylmorphine (5) was prepared from 6-O-acetylmorphine (3) via alkylation with N,N-dimethylformamide di-tert-butyl acetal, followed by hydrolytic removal of the 3-(dimethylamino)-2-propenoate group. The same process was used to prepare the tert-butyl ether of levorphanol (6), (-)-3-tert-butoxy-N methylmorphinan (8). Both 5 and 8 exhibited in vitro affinity for the opiate receptor comparable to codeine and had analgesic properties in the writhing test. Only 5 exhibited activity in the tail-flick procedure and neither compound showed significant antitussive activity. PMID- 6292421 TI - N-Methylnalorphine: definition of N-allyl conformation for antagonism at the opiate receptor. PMID- 6292422 TI - Derivatives of the potent angiotensin converting enzyme inhibitor 5(S)-benzamido 4-oxo-6-phenylhexanoyl-L-proline: effect of changes at positions 2 and 5 of the hexanoic acid portion. AB - Several derivatives of the potent angiotensin converting enzyme inhibitor 5(S) benzamido-4-oxo-6-phenylhexanoyl-L-proline (1) were synthesized and tested for converting enzyme inhibition activity and blood pressure lowering effects in rats. One compound, 5(S)-benzamido-2(R)-methyl-4-oxo-6-phenylhexanoyl-L-proline (2a), had and I50 against angiotensin converting enzyme of 1.0 x 10(-9) M and is the most potent inhibitor prepared thus far in this class of compounds. Testing of 2a orally at 30 mg/kg for inhibition of the angiotensin I induced blood pressure increase in conscious normotensive rats gave 100% inhibition that required 143 min before the angiotensin I blood pressure response returned to 70% of the pretreatment control response. In the conscious renal hypertensive rat, 2a given orally at a dose of 3 mg/kg caused a lowering of blood pressure that reached its maximum of 40 mmHg 8 h following drug administration. PMID- 6292423 TI - Nonsteroidal estrogens: synthesis and estrogen receptor binding affinity of derivatives of (3R*,4S*)-3,4-bis(4-hydroxyphenyl)hexane (hexestrol) and (2R*,3S*) 2,3-bis(4-hydroxyphenyl)pentane (norhexestrol) functionalized on the side chain. AB - A series of nonsteroidal, side-chain functionalized estrogens based on (3R*,4S*) 3,4-bis(4-hydroxyphenyl)hexane (hexestrol) and (2R*,3S*)-2,3-bis(4 hydroxyphenyl)pentane (norhexestrol) has been prepared; these include amide, diazo ketone, ester, alcohol, ketone, fluoro, bromo, iodo, and saturated hydrocarbon derivatives. Analysis of the binding affinity of these compounds to the uterine estrogen receptor, measured by competitive binding assay, reveals trends that can be related to the steric size, the hydrophobicity, and the hydrogen bond accepting character of the side-chain substituents. Comparison of binding affinities between norhexestrol and hexestrol derivatives indicates that, in general, the norhexestrols show significantly higher receptor binding affinities, making this series of compounds ideally suited as functional probes for the estrogen receptor. PMID- 6292424 TI - (2R*,3S*)-1-[125I]Iodo-2,3-bis(4-hydroxyphenyl)pentane ([125I]iodonorhexestrol) and (2R*,3S*)-1-[77Br]Bromo-2,3-bis(4-hydroxyphenyl)pentane ([77Br]bromonorhexestrol), two gamma-emitting estrogens that show receptor mediated uptake by target tissues in vivo. AB - Two gamma-emitting estrogen analogues, (2R*,3S*)-1-[125I]iodo-2,3-bis(4 hydroxyphenyl)pentane ([125I]iodonorhexestrol) and (2R*,3S*)-1-[77Br]bromo-2,3 bis(4-hydroxyphenyl]pentane ([77Br]bromonorhexestrol), have been prepared by halide ion displacement on a labile trifluoromethanesulfonate derivative of a suitably protected precursor, followed by mild acid deprotection. Although halide displacement on a more stable tristrifluoromethanesulfonate derivative was successful, the basic conditions required for deprotection of this precursor resulted in destruction of the products by a base-induced spiroelimination reaction. In immature female rats, both of these halonorhexestrols demonstrated preferential uptake by the uterus that could be blocked selectively by coadministration of a large dose of unlabeled estradiol. In a double label comparison with 16 alpha-[125I]iodo-17 beta-estradiol the uterine uptake of [77Br]bromonorhexestrol was notably less selective. Stability studies in vitro and in vitro have indicated that both iodo- and bromonorhexestrol are quite labile, and this lability compromises the selectivity of their uptake by estrogen target tissues in vivo. p-Hydroxyphenethyl halides are known to be unusually prone to a base-catalyzed solvolysis, via cyclization of the phenolate to a spirocyclohexadienone intermediate. This unusual solvolytic mechanism may contribute to the lability of these halonorhexestrols in vivo. PMID- 6292425 TI - Synthesis and antiviral properties of some 2'-deoxy-5-(fluoroalkenyl)uridines. AB - The following 5-substituted 2,4-dimethoxypyrimidines were synthesized: 5-(2,2,2 trichloro-1-hydroxyethyl), 5-(2,2,2-trichloro-1-fluoroethyl),5-(2,2-dichloro-1 fluorovinyl) (5), and 5-(perfluoropropen-1-yl) (a mixture of E and Z isomers, 6 and 7). Demethylation of 5 gave 5-(2,2-dichloro-1-fluorovinyl)uracil, and demethylation of the mixture of 6 and 7 gave some pure (E)-5-(perfluoropropen-1 yl)uracil. Compound 5 was converted into its 2'-deoxyribonucleoside (12) and its alpha-anomer by standard procedures. 2'-Deoxy-3,5-dilithio-3',5'-O bis(trimethylsilyl)uridine was reacted with the appropriate fluoroalkene to give the following 5-substituted 2'-deoxyuridines in low yield (6-24%): 5-(2-chloro 1,2-difluorovinyl) (a mixture of E and Z isomers, 15 and 16, which were separated on a small scale), 5-(perfluoropropen-1-yl), 5-(perfluorocyclohexen-1-yl), and 5 (perfluorocyclopenten-1-yl). In these reactions, 2'-deoxy-5 (trimethylsilyl)uridine and 2'-deoxyuridine were also formed. The 5-substituted 2'-deoxyuridines were tested for activity against herpes simplex virus type 1. Compound 12 and the mixture of 15 and 16 had an ID50 of 20-26 micrograms/mL in Vero cells. The activity of the mixture resided in one isomer, which by analogY with the corresponding (Z)- and (E)-5-(2-bromovinyl)-2'-deoxyuridines was concluded to be the Z isomer (16). PMID- 6292426 TI - The effect of minocycline on potassium leakage from red cells: a study of the genetics and relationship to vestibular adverse reactions. PMID- 6292428 TI - Purification and characterisation of a fimbrial haemagglutinin from Bordetella pertussis for use in an enzyme-linked immunosorbent assay. AB - The fimbrial haemagglutinin (F-HA) of Bordetella pertussis grown on solid medium was extracted with 1M sodium acetate for 72 h at 20 degree C, and partially purified by Sephacryl S-300 gel chromatography. A pooled fraction with fimbrial haemmagglutinating activity was shown to contain fimbriae haemagglutinating activity was shown to contain fimbriae of the expected morphology by electron microscopy. Chemical and biological assays showed that the F-HA fraction contained some heat-labile agglutinogen and lipopolysaccharide but no measureable lymphocytosis-promoting factor or heat-labile toxin. The F-HA fraction used as antigen in an enzyme-linked immunosorbent assay (ELISA) permitted the detection of antibodies in convalescent serum from a patient with whooping cough. The impurities, heat-labile agglutinogens and lipopolysaccharide, did not contribute to the ELISA activity. The method for preparation of the F-HA antigen is simple, reproducible and gives a high yield. PMID- 6292427 TI - Isolation and characterisation of two strains of Herpesvirus hominis type 1 from fallopian tubes. AB - Herpesvirus hominis type 1 (HSV-1) was isolated from fallopian-tube biopsies of two women who were using an intra-uterine contraceptive device; the strains were isolated on different dates and at different hospitals. Both the isolates were shown to be HSV-1 by restriction-enzyme analysis, polypeptide profiles and indirect immuno fluorescence; the two isolates could not be distinguished by these tests, but the tests distinguished them from HSV-1 strain 71-15 and HSV-2 strain 333. The viruses were again isolated from samples of fallopian-tube tissue stored in liquid nitrogen, and their polypeptide and restriction-enzyme profiles were the same as those in the initial isolates. Virus was not isolated from fallopian-tube samples obtained from other IUD patients undergoing surgery at the same two hospitals at the same time. PMID- 6292429 TI - Detection of antibodies in human serum against the fimbrial haemagglutinin of Bordetella pertussis by enzyme-linked immunosorbent assay. AB - Antibody responses in human sera against Bordetella pertussis during natural infection were investigated by a microplate enzyme-linked immunosorbent assay (ELISA) with a purified fimbrial haemagglutinin preparation as antigen. Significant rises of specific IgG, IgM and IgA were demonstrated in paired sera. A secondary type of antibody response was found in most children and adults. In children, the type of response correlated with previous vaccination status; there was a primary response in unvaccinated children. A survey of antibodies in the general population showed low IgG titres in a small proportion of sera from the youngest healthy children. The titres and the number of individuals with measureable antibodies increased with age. In a limited study of the effect of vaccination, significant rises of titres were demonstrated after vaccination. The ELISA test was specific for antibodies against B. pertussis except that the test also seemed to measure antibody to B. parapertussis. A comparison between ELISA and the complement-fixation test showed a good correlation between the tests only in sera from children 1-12 years old. PMID- 6292430 TI - Uptake of arachidonic acid into membrane phospholipids: effect on chloride transport across cornea. AB - We demonstrate that arachidonic acid (AA) stimulation of chloride transport across frog cornea is mediated via two independent pathways: (1) stimulation of prostaglandins and cAMP synthesis, and (2) a direct physical change in the membrane produced by substitution of different phospholipid acyl chains. AA is well known as a precursor in the synthesis of prostaglandins, which have been shown to stimulate cAMP synthesis and chloride transport in frog cornea. We show that frog cornea can convert exogenous AA to PGE2, but that in the presence of 10(-5) M indomethacin both the conversion to PGE2 and stimulation of cAMP are completely blocked. However, with indomethacin the action of AA to stimulate chloride transport (as measured by SCC) remains, but peak height of the response is reduced to 57% of that found when AA alone is given. Similarly, we show that propranolol completely blocks cAMP stimulation, but stimulation of SCC is reduced to 45% of the original response. Therefore, cAMP appears to be responsible for roughly half of the observed stimulation in SCC. By gas chromatographic analysis we show that significant quantities of AA can rapidly substitute into membrane phospholipids of corneal epithelium and L929 cells following the addition of AA to the medium. Modification of membrane phospholipid structure can affect membrane viscosity, membrane-bound enzyme activity, and the distribution and lateral mobility of integral proteins. It seems likely that such alterations in the properties of the membrane may modulate the rate of chloride transport, and this may constitute the second mechanism. Upon addition of AA, both mechanisms appear to stimulate chloride transport simultaneously, and are apparently additive. We show that prolonged exposure to AA results in a large incorporation of AA into phospholipid and consequently, a perturbation in the ratio of unsaturated to saturated fatty acids. We also find evidence of a compensatory cellular mechanism that alters the ratio of endogenously synthesized fatty acids and tends to reduce the membrane-perturbing effect of AA.U PMID- 6292431 TI - Noise analysis reveals K+ channel conductance fluctuations in the apical membrane of rabbit colon. AB - In this paper we describe current fluctuations in the mammalian epithelium, rabbit descending colon. Pieces of isolated colon epithelium bathed in Na+ or K+ Ringer's solutions were studied under short-circuit conditions with the current noise spectra recorded over the range of 1-200 Hz. When the epithelium was bathed on both sides with Na+ Ringer's solution (the mucosal solution contained 50 microM amiloride), no Lorentzian components were found in the power spectrum. After imposition of a potassium gradient across the epithelium by replacement of the mucosal solution by K+ Ringer's (containing 50 microM amiloride), a Lorentzian component appeared with an average corner frequency, fc = 15.6 +/- 0.91 Hz and a mean plateau value So = (7.04 +/- 2.94) x 10(-20) A2 sec/cm2. The Lorentzian component was enhanced by voltage clamping the colon in a direction favorable for K+ entry across the apical membrane. Elimination of the K+ gradient by bathing the colon on both sides with K+ Ringer's solutions abolished the noise signal. The Lorentzian component was also depressed by mucosal addition of Cs+ or tetraethylammonium (TEA) and by serosal addition of Ba2+. The one-sided action of these K+ channel blockers suggests a cellular location for the fluctuating channels. Addition of nystatin to the mucosal solution abolished the Lorentzian component. Serosal nystatin did not affect the Lorentzian noise. This finding indicates an apical membrane location for the fluctuating channels. The data were similar in some respects to K+ channel fluctuations recorded from the apical membranes of amphibian epithelia such as the frog skin and toad gallbladder. The results are relevant to recent reports concerning transcellular potassium secretion in the colon and indicate that the colon possesses spontaneously fluctuating potassium channels in its apical membranes in parallel to the Na+ transport pathway. PMID- 6292432 TI - 5' termini of polyoma virus early region transcripts synthesized in vivo by wild type virus and viable deletion mutants. PMID- 6292433 TI - Determination of sequences at the capped 5' ends of polyoma virus early region transcripts synthesized in vivo and in vitro demonstrates an unusual microheterogeneity. PMID- 6292434 TI - IS50-mediated inverse transposition. Discrimination between the two ends of an IS element. PMID- 6292435 TI - Structure and stability of transposon 5-mediated cointegrates. PMID- 6292436 TI - Amplification and expression of sequences cotransfected with a modular dihydrofolate reductase complementary dna gene. PMID- 6292437 TI - Permissive effect of glucocorticoids in catecholamine action in the heart: possible mechanism. PMID- 6292439 TI - A helpful hint for chest radiology: "look behind the heart". AB - Pathologic abnormalities on frontal chest radiographs are often located solely or partially in the retrocardiac region. It is essential to include a look behind the heart when viewing the frontal x-ray. Cases of pneumonia, metastases, and posterior mediastinal mass, all of which would have been missed without a look behind the heart, are used to illustrate this precept. PMID- 6292438 TI - Characterization of the GTP-binding component of the adenylate cyclase system in isolated myocardial muscle cells. PMID- 6292440 TI - Cell surface-mediated cellular interactions: effects of B104 neuroblastoma surface determinants on C6 glioma cellular properties. AB - To study the influence of cell surface-associated molecules on intercellular communication, C6 glioma cells were cultured both on plastic and on substrata of paraformaldehyde-fixed B104 neuroblastoma cells. By then comparing the phenotypic expression of these "cocultured" C6 cells with cells cultured on tissue culture plastic, the influence of the cellular substratum was determined. The beta adrenergic-responsive cyclic AMP-generating system of C6 cells was compared on these various substrata. We found that fixed beds of dibutyryl cyclic AMP (dbcAMP)-treated B104 cells uncoupled beta-receptors from adenylate cyclase, whereas fixed beds of similarly treated C6 cells did not. However, other cellular properties were not affected by growth atop fixed dbcAMP-treated B104 cell beds including the rate of C6 cellular proliferation and their rate of protein synthesis. The cell surface-associated determinant on B104 cells capable of uncoupling the beta-responsive cyclase system of C6 cells is probably a protein, as judged by its susceptibility to protease treatment. Other properties of C6 cells were also affected by the various substrata including basal and hydrocortisone-induced levels of glycerol phosphate dehydrogenase (GPDH; an oligodendroglial marker) and the rate of RNA synthesis in these cells. PMID- 6292441 TI - Sequestration requirements for the degradation of 125I-labeled beta nerve growth factor bound to embryonic sensory neurons. AB - Nerve growth factor interacts with responsive cells by binding to cell surface membrane receptors. There are two different receptors on both embryonic sensory and sympathetic neurons, a high-affinity (type I) receptor and a lower-affinity (type II) receptor. Sequestration, which we have defined as bound nerve growth factor that becomes inaccessible to the external milieu with time, occurs through the type I receptor on both sensory and sympathetic neurons. We describe here a process subsequent to sequestration involving internalization and degradation of bound nerve growth factor and showing that bound nerve growth factor is not degraded under the following conditions: (1) low temperature, ie 4 degrees C; (2) when a large excess of unlabeled nerve growth factor is added concomitantly with the labeled nerve growth factor and the temperature is raised from 4 degrees C to 37 degrees C; (3) when metabolic inhibitors sodium fluoride and dinitrophenol are added concomitantly with the labeled nerve growth factor and the temperature is raised from 4 degrees to 37 degrees C. On the other hand, conditions that allow bound nerve growth factor to be degraded are the following: (1) incubation of the sensory nerve cells at low temperature (ie, 4 degrees C) only in the presence of labeled nerve growth factor, then raising the temperature to 37 degrees C; (2) when sodium fluoride and dinitrophenol are added when the temperature is raised to 37 degrees C; (3) when excess unlabeled nerve growth factor is added when the temperature is raised to 37 degrees C. These studies are consistent with the idea that nerve growth factor has to bind to the cells in order to be degraded; however, binding is not sufficient for degradation to occur. Second, the bound nerve growth factor must be sequestered in order to be degraded. Third, the process of internalization of the bound nerve growth factor, unlike sequestration, is not an energy-dependent process. Thus, it seems reasonable to suggest the following steps for the interaction of nerve growth factor with responsive cells: binding to a cell surface membrane receptor, followed by sequestration of the bound nerve growth factor, and finally, internalization of the sequestered nerve growth factor. PMID- 6292442 TI - Structural features of a specialized region of the epididymis of the Siberian hamster (Phodopus sungorus). AB - Light and electron microscopic observations of zone six of the Siberian hamster (Phodopus sungorus) epididymis reveal that two distinct tubule populations exist in this region. The population of smaller tubules apparently arises as diverticula of the larger ducts but, though contiguous, the contents of each remain different and separate. While the larger tubules are laden with sperm and seminal fluid, the smaller ones area filled with degenerating cells and cellular debris. Cytologically, the features of the two tubule types are similar. Both display a prominent Golgi apparatus, substantial amounts of rough endoplasmic reticulum, an extensive population of various lysosomal forms and scattered crystalline inclusions. PMID- 6292443 TI - The effect of magnesium trisilicate and kaolin on the in vivo absorption of chloroquine. AB - An in vitro study indicated that certain antacids and adsorbents may decrease the oral availability of th two widely used antimalarial agents chloroquine and pyrimethamine. To determine if this data was applicable to the clinical (in vivo) situation, plasma levels of one of the antimalarial agents (chloroquine) were followed in six Negro--Arab volunteers both when given alone and when taken with separate doses of two of the implicated interactants (magnesium trisilicate and kaolin). This in vivo work confirmed the in vitro findings; chloroquine area under the plasma concentration-time curve data were decreased by both magnesium trisilicate (18.2%) and kaolin (28.6%). Similar results could be expected for pyrimethamine. It is suggested therefore, to avoid loss of drug, that the antimalarials should not be taken with gastrointestinal medications of this type or that their administration should be separated by at least 4 h to reduce the risk of them interacting in the gut, thus preventing drug adsorption to the antacids/adsorbents, and loss of systemic availability. PMID- 6292444 TI - Radionuclide salivary scanning. PMID- 6292445 TI - Regulation of alpha genes of herpes simplex virus: the alpha 27 gene promoter thymidine kinase chimera is positively regulated in converted L cells. AB - In cells infected with herpes simplex virus 1, the expression of viral genes is coordinately regulated and sequentially ordered; the alpha genes are expressed first and are followed by beta and gamma genes in a cascade fashion. Earlier, this laboratory reported (Post et al., Cell 24:555-565, 1981) that a chimeric gene, constructed by the replacement of 50 base pairs of DNA coding for 5' nontranslated leader and sequences upstream of the site of transcription initiation of thymidine kinase (a beta gene) by corresponding sequences of alpha gene no. 4, was regulated as an alpha gene. Of particular interest was the observation that in cells converted to TK(+) phenotype, the chimeric gene was positively regulated by superinfecting virus. In this paper, we report two series of experiments. First, we determined that transcription of alpha gene no. 27 is initiated at or near a five-nucleotide sequence flanked by an eight-nucleotide perfect inverted repeat situated from 256 to 277 bases to the right of the left terminus of the BamHI B fragment. In the second series of experiments, we constructed a chimeric gene which consisted of the thymidine kinase sequences described above but was fused to a DNA fragment expected to contain the promoter regulatory region of alpha gene 27 and stretching from approximately -270 to the +55 nucleotide. The chimeric gene in converted cells was amplified upon superinfection with TK(-) virus only when the promoter-regulatory region was in the correct transcriptional orientation relative to the leader and structural sequences of the thymidine kinase gene. The requirements for amplification of the expression of this chimeric thymidine kinase gene were exactly the same as those previously reported for the alpha gene no. 4-thymidine kinase chimera and different from those of the standard (beta) thymidine kinase. We conclude that the positive regulation of expression of alpha gene no. 4 deduced in previous studies may be a general property of alpha genes and that the promoter-regulatory region of alpha gene no. 27 is within a sequence contained between -270 and +55 nucleotides relative to the transcription initiation site. PMID- 6292446 TI - In vitro transcription of two human rotaviruses. AB - The RNA polymerase activities of a cultivatable (Wa) and a noncultivatable (DS-1) strain of human rotavirus were studied. Under optimal conditions, transcription of all of their RNA segments occurred, as evidenced by the hybridization of labeled transcripts to genomic RNA. Cross-hybridization between the two viruses showed that none of their 11 genes were completely homologous. The transcription products could be translated in vitro, yielding proteins with an electrophoretic pattern resembling that obtained with proteins labeled in vivo during infection with the Wa virus. PMID- 6292447 TI - Identification of ecotropic proviral sequences in high- and low-ecotropic-virus producing mouse strains. AB - The arrangement of endogenous ecotropic retroviruses in selected high- and low ecotropic-virus-producing mouse strains was examined by Southern blot hybridization analysis, using an ecotropic retrovirus-specific DNA probe. High ecotropic-virus-producing mouse strains of the AKR family displayed heterogeneity with respect to the number of copies and the sites of insertion of endogenous ecotropic specific DNA. This diversity was seen even among individuals of the same AKR subline. Contrastingly, individuals within the same low-ecotropic retrovirus-producing mouse strain showed no evidence of variability in their endogenous ecotropic proviral sequences. These results favored the hypothesis that germ line proviral reinsertion was responsible for the proviral sequence heterogeneity observed in high-ecotropic-virus-producing mouse strains. PMID- 6292448 TI - In situ hybridization: general infectivity assay for retroviruses. AB - We have devised a general infectivity assay for retroviruses. A virus-specific [32P]DNA probe is hybridized in situ to a monolayer culture, and foci of infected cells in the monolayer are detected by exposure of the hybridized culture to X ray films. The method is quantitative, in that it gives the same titer for Moloney murine leukemia virus as does the standard UV-XC test. The specificity of the assay is indicated by the fact that murine leukemia virus and baboon endogenous virus do not cross hybridize under the conditions used. The assay is completed within 1 to 3 weeks and should be broadly applicable for retroviruses which replicate without altering cellular morphology: its use is demonstrated with mouse mammary tumor virus and the helper virus of the reticuloendotheliosis complex. PMID- 6292449 TI - Infectivity and glycoprotein processing of herpes simplex virus type 1 grown in a ricin-resistant cell line deficient in N-acetylglucosaminyl transferase I. AB - We report on the replication of herpes simplex virus type 1 (HSV-1) and viral glycoprotein processing in RicR14 cells, a mutant ricin-resistant cell line defective in N-acetylglucosaminyl transferase I activity. In these cells HSV 1(MP) and (F) replicated to yields very similar to those in parental BHK cells. The kinetics of HSV-1 adsorption in mutant and in parent cells was also essentially identical. Progeny virions from ricin-resistant and wild-type cells displayed comparable specific infectivities. However, in the mutant cells the efficiency of plating of progeny virus from both RicR14 and BHK cells was reduced. HSV-1(MP) failed to induce syncytia in RicR14 cells either in a plaque assay or after a high-multiplicity infection. Moreover, the fully glycosylated forms of glycoproteins (gB, gC, and gD) were totally absent, and only the partially glycosylated precursors (pgC, pgD. and a triplet in the gB-gA region) accumulated in HSV-1-infected ricin-resistant cells and in herpesvirions made in these cells. Consistent with these results analysis of pronase glycopeptides from cells labeled with [14C]glucosamine showed a strong decrease of sialylated complex-type oligosaccharides and a dramatic accumulation of the neutral mannose rich chains. The latter chains predominate in partially glycosylated precursors, whereas the complex acidic chains predominate in the fully processed forms of HSV glycoproteins. These results taken together indicate that (i) host-cell N acetylglucosaminyl transferase I participates in the processing of HSV glycoproteins; and (ii) infectivity of herpesvirions does not necessarily require the mature form of gB. The absence of HSV-1(MP)-induced fusion in RicR14 cells is discussed. PMID- 6292450 TI - Polyoma virus mutant with normal transforming ability but impaired tumorigenic potential. AB - Cloned DNA from the P155 mutant of polyoma virus transforms cells in culture as efficiently as wild-type DNA, but has a much lower tumorigenic potential when injected into newborn rats. Like cells transformed by wild-type DNA, cells transformed by the mutant DNA grow in low serum concentrations, form colonies in agar suspension, and grow to high saturation densities compared with untransformed cells. They are, however, much less tumorigenic since they transplant 100- to 2,000-fold less efficiently than cells transformed by wild type DNA. Substitution of the region between 89.7 and 1.8 map units by the corresponding region of P155 DNA decreased the tumorigenicity of wild-type DNA. When this region was isolated from wild-type DNA and substituted in P155 DNA, the tumorigenicity of the latter increased to values comparable to those of wild-type DNA. This showed that the lesion affecting tumorigenicity occurred between 89.7 and 1.8 map units on the polyoma virus genome. Sequence analysis in this region revealed a 12-base-pair deletion between nucleotides 1,347 and 1,360. This identified P155 as an mlt mutant, i.e., a mutant with a deletion from a region which encodes parts of the large and middle T antigens. PMID- 6292451 TI - Effect of tunicamycin on rotavirus assembly and infectivity. AB - Bovine rotavirus grown in the presence or absence of tunicamycin was analyzed with respect to yield of infectious virus, the ratio of complete to incomplete particles, and polypeptide composition. Tunicamycin at a concentration of 1 microgram/ml reduced virus yields by 4 logs and completely prevented the incorporation of [3H]uridine into complete rotavirus particles, as determined by cesium chloride gradient analysis. Concomitant with a reduction in complete particles, three rotavirus polypeptides shifted in their relative position on polyacrylamide gels from 41,900-molecular-weight position (41.9K), 29.3K, and 16.1K to migrate at 35.5K, 22.7K, and 15.5K, respectively. Limited proteolysis indicated that the lower-molecular-weight polypeptides possessed the same constituent peptides as the larger polypeptides, suggesting that they represented the unglycosylated equivalents. These results suggest that interference with glycosylation prevents proper assembly of the outer coat proteins in bovine rotavirus. PMID- 6292452 TI - Expression of cloned herpesvirus genes. I. Detection of nuclear antigens from herpes simplex virus type 2 inverted repeat regions in transfected mouse cells. AB - Three different recombinant plasmids containing the entire 15-kilobase L and S inverted repeat sequence of herpes simplex virus type 2 DNA have been introduced into cultured Ltk- or BSC cells by both the calcium and DEAE-dextran transfection procedures. In each case, after 24 h approximately 1% of the cells gave strongly positive nuclear staining when assayed by immunofluorescence with hyperimmune antisera made against early and immediate-early infected-cell polypeptides. The nuclear fluorescence pattern and intensity mimicked that observed within 2 to 3 h after infection of Ltk- cells with either herpes simplex virus type 1 or type 2 wild-type virus. Herpes simplex virus type 1 (KOStsB2)-infected Ltk- cells under nonpermissive conditions did not express these antigens in the nucleus. Therefore, we conclude that either one or both of the 185,000- and 110,000 molecular-weight immediate early proteins, or some other as yet unknown gene product encoded entirely within the inverted repeats, can be transiently expressed in large amounts in transfected cells in the absence of other viral genes or accompanying virion components. Permanent mouse cell lines derived from transfection with these plasmids by using the thymidine kinase coselection procedure did not express sufficient nuclear antigen to be detectable by immunofluorescence. PMID- 6292453 TI - Monensin inhibits the processing of herpes simplex virus glycoproteins, their transport to the cell surface, and the egress of virions from infected cells. AB - HEp-2 cells or Vero cells infected with herpes simplex virus type 1 were exposed to the ionophore monensin, which is thought to block the transit of membrane vesicles from the Golgi apparatus to the cell surface. We found that yields of extracellular virus were reduced to less than 0.5% of control values by 0.2 microM monensin under conditions that permitted accumulation of cell-associated infectious virus at about 20% of control values. Viral protein synthesis was not inhibited by monensin, whereas late stages in the post-translational processing of the viral glycoproteins were blocked. The transport of viral glycoproteins to the cell surface was also blocked by monensin. Although the assembly of nucleocapsids appeared to be somewhat inhibited in monensin-treated cells, electron microscopy revealed that nucleocapsids were enveloped to yield virions, and electrophoretic analyses showed that the isolated virions contained immature forms of the envelope glycoproteins. Most of the virions which were assembled in monensin-treated cells accumulated in large intracytoplasmic vacuoles, whereas most of the virions produced by and associated with untreated cells were found attached to the cell surface. Our results implicate the Golgi apparatus in the egress of herpes simplex virus from infected cells and also suggest that complete processing of the viral envelope glycoproteins is not essential for nucleocapsid envelopment or for virion infectivity. PMID- 6292454 TI - Purification and characterization of bovine rotavirus cores. AB - Using the chaotropic effect generated by a high concentration of CaCl2, we converted calf rotavirus particles into cores of 40 nm in diameter. These cores were purified by rate zonal centrifugation in sucrose gradients and by isopycnic gradients. They had a sedimentation coefficient of 280S +/- 20S and a density of 1.44 g/ml in CsCl. When analyzed by polyacrylamide gel electrophoresis, they contained three polypeptides (VP125, VP89, and VP78). The major internal polypeptide of the virion (VP39) was recovered in a purified and soluble form in the top fractions of the sucrose gradients. From this stepwise degradation, it appears that VP39 is the most external polypeptide of dense particles. In contrast to reovirus cores, calf rotavirus cores did not exhibit transcriptase activity. Purified VP39 also did not exhibit transcriptase activity when tested after being mixed with purified rotavirus genome RNA as a template. Transcriptase activity was partially recovered when ionic conditions were adjusted to permit the reassociation of VP39 with the cores. PMID- 6292455 TI - Characterization of a cell culture persistently infected with the DA strain of Theiler's murine encephalomyelitis virus. AB - We established a persistent infection in L 929 cells with the DA strain of Theiler's murine encephalomyelitis virus. Our studies showed that only a small number of cells in the cultures contained infectious virus or viral antigen. A role for interferon in the maintenance of persistence was suggested. Viral isolates from the cultures were not temperature sensitive, nor did they contain viral capsid polypeptide mutations or defective interfering particles. T1 oligonucleotide maps showed evidence of mutation in two of three isolates. PMID- 6292456 TI - Detailed characterization of an apparently unspliced beta herpes simplex virus type 1 gene mapping in the interior of another. AB - We precisely localized the coding region and determined the nucleotide sequence of a 1.2-kilobase beta herpes simplex virus type 1 mRNA which underlies the 3' region of the 5.2-kilobase beta mRNA mapping in HindIII fragment K. This mRNA, which lacks readily detectable splices, has its own promoter by the criteria of identification of putative herpes simplex virus type 1 control sequences and in vitro transcription by a Manley polymerase system. PMID- 6292457 TI - Hypomethylation of Epstein-Barr virus DNA in the nonproducer B-cell line EBR. AB - The conversion of the Epstein-Barr virus-negative Ramos cell line has previously been shown to result in an Epstein-Barr virus-positive non-virus-producer cell line, EBR. We report here that Epstein-Barr virus DNA from EBR alone among several cell lines examined was totally unmethylated at three of four sites containing guanine plus cytosine which were tested. This is in direct contrast to reports of high degrees of methylation in the DNAs of other animal viruses, including herpesviruses, isolated from cells in which the viral genome is expressed at a low level. PMID- 6292458 TI - Is progressive multifocal leukoencephalopathy a chronic disease because of defective interfering particles or temperature-sensitive mutants of JC virus? AB - JC virus was examined for temperature sensitivity and for evidence of defective interfering particles as a means of explaining the slow chronic nature of progressive multifocal leukoencephalopathy (PML). JC virus direct from the brain tissue of seven persons with PML was not temperature sensitive as indicated by in vitro assay at 37 and 39 degrees C. In fact, more cells contained viral antigen at 39 than at 37 degrees C. The amount of infectious virus also was increased at 39 degrees C. Virions isolated directly from diseased brain tissue had a higher buoyant density than did virus from the same PML patient passaged in culture and containing genomic deletions. In contrast to DNA from culture-passed virus, DNA extracted from virions direct from brain tissue was homogeneous in length. In 13 separate cases examined, the viral DNA direct from the brain was homogeneous although variations in length were noted among DNAs from different cases. Restriction enzyme cleavage patterns identified all as JC virus DNA. It was concluded that neither temperature sensitivity nor DI particles can be used to explain the slow, progressive nature of PML. PMID- 6292459 TI - Phosphorylation pattern of large T antigens in mouse cells infected by simian virus 40 wild type or deletion mutants. AB - The phosphorylation sites of simian virus 40 (SV40) large tumor (T) antigens have been extensively studied in productive infection of monkey cells. In this study, we analyzed the phosphorylation sites of large T antigen from SV40-infected nonpermissive mouse cells by partial proteolysis fingerprints and analysis of the phosphoamino acids present in the resulting fragments. The wild-type virus and deletion mutants (dl1263, dl1265, dl2194, and dl2198) were used for infection. On the basis of our results and published data (M. Schwyzer, R. Weil, and H. Zuber, J. Biol. Chem. 225:5627-5634, 1980), a cleavage map of large T antigen was established. It was reported that at least four sites of phosphorylation were present. The amino-terminal part of the molecule contained both phosphoserine and phosphothreonine. One phosphothreonine residue was located in the prolinerich C terminal end of the molecule at position 701 or 708. On the basis of the concensus as to the amino acid sequence surrounding the recognition sites for protein kinases, it was possible to more precisely locate this phosphothreonine at residue 701. Moreover, the C-terminal part of the molecule contained phosphoserine at a more internal position. In addition, this study firmly established the presence of a phosphothreonine in the N-terminal part of large T antigen. In conclusion, it was shown that the location of phosphorylation sites of large T antigen produced by nonpermissive mouse cells infected by SV40 is strikingly similar to that reported by other groups for large T antigen produced by SV40-infected permissive cells. PMID- 6292460 TI - Immunization against the polyoma virus-induced tumor-specific transplantation antigen by early region mutants of the virus. AB - To investigate the relation between the polyoma tumor-specific transplantation antigen and the virus-coded proteins, mice were immunized by inoculation of a variety of viable polyoma virus mutants and then challenged with polyoma virus induced tumors. Two classes of early region mutants were used. One class produces a normal small T-antigen and truncated middle and large T-antigens. The second class (hr-t mutants) forms a normal large T-antigen together with N-terminal fragments of small and middle T-antigens. All mutants, transforming as well as nontransforming, induced protection against polyoma virus tumors. However, there were quantitive differences between the mutants. The finding that an hr-t mutant could induce tumor rejection suggests that full-length middle and small T antigens are not necessary for the induction of this response. Since intact middle T-antigen is the only virus-coded protein known to associate with the plasma membrane, the possibility must be considered that the polyoma virus tumor specific transplantation antigen consists of cellular components. PMID- 6292461 TI - Cold-sensitive growth of simian virus 40 in semipermissive variants of CV1 cells. AB - Two cell clones were isolated from the simian line CV1, permissive for simian virus 40 (SV40), by selection at low temperature with the tsA239 mutant of SV40. These clones exhibited cold-sensitive semipermissivity to both SV40 virions and SV40 DNA. On the basis of virus yields, their resistance to viral DNA was increased approximately 15 times over that of CV1 cells when the incubation temperature was lowered from 38.5 to 33.5 degrees C. A further 30- to 40-fold resistance increase was exhibited at both temperatures upon infection with SV40 virions. Partial characterization of these clones indicated that the cold sensitivity affected an early function in viral growth, between viral uncoating and the appearance of T-antigen positivity, with a burst-size decrease in all cells at the restricted temperature. This conditional defect appeared to be superimposed upon a temperature-independent uncoating defect, presumably carried in a CV1 subpopulation from which the two clones were ultimately selected. PMID- 6292462 TI - Physical and biological features of polyoma virus mutants able to infect embryonal carcinoma cell lines. AB - Three new polyoma mutants were selected for their ability to grow on the embryonal carcinoma cell line F9. These mutants share in common an insertion of two nucleotides, a thymine and an adenine, in the noncoding region located on the late side of the origin of replication. We have found that these insertions exist in all of the other polyoma virus mutants able to grow on F9 cells (Fujimura et al., Cell 23:809-814, 1981; Katinka et al., Nature (London) 290:720-722, 1981; K. Sekikawa and A. J. Levine, Proc. Natl. Acad. Sci. U.S.A. 78:1100-1104, 1981). The region containing these insertions could be folded into a stable secondary structure which included a guanine plus cytosine (G + C)-rich stem. The adenine and thymine were inserted in such a way that they maintained the palindrome in the G + C-rich stem and were complementary in the putative secondary structure that we present here. Another class of polyoma virus mutants selected on a multipotential carcinoma cell line (PCC4-Aza) were characterized by a more complex rearrangement (deletion and duplication) which occurred in the same region. This arrangement preserved the G + C-rich palindrome and also yielded a sequence which still allowed the folding of another type of stable secondary structure. The significance of these findings is discussed. PMID- 6292463 TI - Acquisition of proviral DNA of mouse mammary tumor virus in thymic leukemia cells from GR mice. AB - Male mice of strain GR develop T-cell leukemia at a low frequency late in life. These leukemia cells contain large amounts of mouse mammary tumor virus (MMTV) RNA and MMTV proteins in a precursor form (Nusse et al., J. Virol. 32:251-258, 1979). We used restriction enzyme analysis and molecular hybridization to identify MMTV proviruses in the DNA of these leukemia cells. GR leukemia cells contained additional integrated MMTV proviruses at various sites in the genome. This amplification of MMTV proviruses in GR leukemia cells is not restricted to one particular endogenous MMTV provirus of strain GR. The number and location of the extra MMTV proviruses present in transplants of GR leukemia cells did not change upon serial transplantation of the leukemia cells. Acquisition of MMTV proviruses was also found in a similar leukemia, L1210 of the DBA/2 mouse strain, but not in three other leukemias, SL2 of DBA/2, BW5147 of AKR, and a spontaneous thymoma of BALB/c. The two main classes of MMTV RNA, 35S and 24S, were present in the cytoplasmic RNA of GR leukemia cells, indicating that the aberrant processing of MMTV precursor proteins is not due to anomolously sized RNAs. We could not detect extra RNAs in GR leukemia cells which would represent read-through transcripts of cellular genes adjacent to the extra MMTV proviruses, initiated by a promoter signal in the right MMTV long terminal repeat sequence. These data suggest that acquisition of MMTV proviruses may coincide with the onset of leukemogenesis in GR male mice. PMID- 6292464 TI - Simian virus 40 encapsidation: characterization of early intermediates. AB - Simian virus 40 chromosomes were separated into various species by a two-step purification consisting of low-ionic-strength glycerol gradient sedimentation followed by low-ionic-strength agarose gel electrophoresis. For each species of simian virus 40 chromosome purified, the comigrating DNA and proteins were identified by agarose or polyacrylamide gel electrophoresis, respectively. Two species of chromosomes were identified which contained form I and form II DNA and large amounts of viral protein; they migrated more slowly than most of the free simian virus 40 chromosomes, which contained very little viral protein. The nuclease susceptibility of these chromosomes suggests to us that they are intermediates in encapsidation, and we describe an encapsidation model. PMID- 6292466 TI - Saturable binding sites for vesicular stomatitis virus on the surface of Vero cells. AB - The binding of vesicular stomatitis virus (VSV) to Vero monkey cells was studied by using virus metabolically labeled with [35S]methionine. Under conditions where viral uptake did not occur (4 degrees C), apparent binding equilibrium was achieved within 12 h at a level representing 12% of the input virus. Two distinct forms of virus-cell interaction were found. At low concentrations of VSV, corresponding to multiplicities used for tissue culture studies, saturable binding was the major form of interaction. Saturation was complete at approximately 4,000 VSV virions per cell. At higher virus concentrations, nonsaturable binding prevailed. Trypsin treatment of Vero cells did not decrease the binding of VSV to the saturable binding sites. Internalization of VSV at 37 degrees C also displayed a saturable component which was directly comparable to that observed for binding. VSV binding to high-affinity, saturable sites on the plasma membrane may represent a receptor-mediated route of viral uptake. PMID- 6292465 TI - Physical mapping of hybrid bacteriophage T7/T3 RNA polymerase genes. AB - The late regions of the T7 and T3 bacteriophage genomes are transcribed by phage specified RNA polymerases, the products of gene 1. Although these phage transcriptional systems share many characteristics and are obviously related, they have diverged to such an extent that neither of their respective RNA polymerases utilizes the promotor sites of the other phage at an appreciable rate. However, it is possible to construct viable T7/T3 hybrids which have hybrid gene 1 sequences; the resultant hybrid enzymes exhibit altered transcriptional patterns in that they are capable of transcribing both T7 and T3 DNA to various degrees. The aim of this study was to define more closely the region(s) of the gene 1 sequence which encodes the transcriptional selectivity determinant by correlating the genetic constitution of these hybrid gene 1 sequences with their transcriptional properties. The recombinant sites within the gene 1 regions of several T7/T3 hybrids were mapped by using restriction sites as genetic markers. The results indicated that forcing a crossover event within a particular region often results in the inadvertant selection of additional genetic rearrangements. Several of the hybrid gene 1 sequences were found to have resulted from multiple crossover events, even though only one was directly selected for. In some cases the predicted crossovers were not detected; instead, several hybrids contained recombination sites elsewhere in the gene 1 region. These findings suggest that only certain combinations of T7/T3 gene 1 sequences are compatible; it may be that active hybrid T7/T3 gene 1 sequences rarely result from single genetic rearrangements. Taken together, the results of this study suggest that more than one region of the gene 1 sequence is involved in transcriptional selectivity. More specifically, the region from approximately 25 to 59% (from the left of the gene), together with the carboxyl end, appears to play an important role. PMID- 6292468 TI - Asynchronous bidirectional replication of polyoma virus DNA. AB - The structure of polyoma virus replicative intermediates isolated from infected 3T6 cells was analyzed by two-dimensional agarose gel electrophoresis (Sundin and Varshavsky, Cell 21:103-114, 1980) and quantitative electron microscopy (Krauss and Benbow, J. Virol. 38:815-825, 1981). DNA replication was initiated at a single site (ori) in essentially all of the replicative intermediates. Most of the early replicative intermediates were formed by unidirectional synthesis in the direction of early transcription. Most mid- and late replicative intermediates contained two replication forks which had traveled unequal distances from the origin. Asynchronous initiation of the two growing forks was postulated to account for these observations. PMID- 6292467 TI - Mouse mammary tumor virus proviral sequences congenital to C3H/Sm mice are differentially hypomethylated in chemically induced, virus-induced, and spontaneous mammary tumors. AB - C3H/Sm mice have lost the exogenous milk-borne mouse mammary tumor virus (MMTV) characteristic of the C3H strain and have a very low (1.5%) incidence of spontaneous mammary tumors, yet they are highly susceptible to mammary carcinogenesis by either chemical carcinogens or infection with the milk-borne virus. We have analyzed the MMTV proviral DNA content of normal tissues and of spontaneous, virus-induced, and chemically induced mammary tumors by restriction endonuclease digestion and Southern blot analysis. Although the results clearly showed additional MMTV sequences in the virus-induced tumor which are not present in normal liver DNA, none of the spontaneous or chemically induced tumors could be shown to contain either newly acquired exogenous or amplified endogenous MMTV sequences. Interestingly, mammary tumors arising in C3H/Sm mice treated simultaneously with infectious MMTV (C3H) and dimethylbenz[a]anthracene (DMBA) possessed new exogenous MMTV DNA even though no quantitative change in tumor production was observed when these mice were compared with C3H/Sm mice treated with DMBA alone (Smith et al., Int. J. Cancer 26:373-379, 1980). Our data indicate that the endogenous MMTV proviral units are extensively methylated in normal tissues, such as livers and normal nonlactating mammary glands. In the absence of MMTV (C3H), we found that in the rare, spontaneously occurring C3H/Sm mammary tumors, certain endogenous MMTV sequences were specifically hypomethylated. Hypomethylation of endogenous MMTV sequences was also noted in the chemically induced mammary tumors, even though radioimmune competition assays for MMTV gp52 and p28 are negative (Smith et al., Int. J. Cancer 27:81-86, 1981). Our results support the conclusion that amplification of endogenous MMTV sequences is not intrinsic to C3H/Sm mouse mammary tumors arising spontaneously or after induction by chemicals. On the other hand, integration of exogenous MMTV DNA into the genome was a constant feature of mammary tumors developing in MMTV (C3H)-infected C3H/Sm mice, even when DMBA was used as the carcinogen. Hypomethylation of some endogenous MMTV sequences is characteristic of C3H/Sm mammary tumors, whether spontaneous or induced by chemicals, which suggests that these sequences are located in actively transcribing regions of the tumor cell genome. PMID- 6292470 TI - Molecular cloning of a family of retroviral sequences found in chimpanzee but not human DNA. AB - A number of retrovirus-like sequences have been cloned from chimpanzee DNA which constitute the chimpanzee homologs of the endogenous colobus type C virus CPC-1. One of the clones contains a nearly complete viral genome, but others have sustained deletions of 1 to 2 kilobases in the polymerase gene. The pattern of related sequences detected in other primate species is consistent with the genetic transmission of these sequences for millions of years. However, the appropriately related sequences have not been detected in human, gibbon, or orangutan DNAs. These results suggest either that this family of sequences has been deleted from humans, gibbons, and orangutans, or that the genes were recently acquired in the chimpanzee and gorilla lineages. PMID- 6292469 TI - Cell-free translation of murine coronavirus RNA. AB - The coding assignments of the intracellular murine hepatitis virus-specific subgenomic RNA species and murine hepatitis virion RNA have been investigated by cell-free translation. The six murine hepatitis virus-specific subgenomic RNAs were partially purified by agarose gel electrophoresis and translated in an mRNA dependent rabbit reticulocyte lysate, and the cell-free translation products were characterized by gel electrophoresis, immunoprecipitation, and tryptic peptide mapping. These studies have shown that RNA 7 codes for the nucleocapsid protein, RNA 6 codes for the E1 protein, RNA 3 codes for the E2 protein, and RNA 2 codes for a 35,000-dalton nonstructural protein. Genomic RNA directs the cell-free synthesis of three structurally related polypeptides of greater than 200,000 in molecular weight. PMID- 6292471 TI - Characterization of c-lil, a chicken cellular sequence associated with a stock of B77 avian sarcoma virus. AB - Using biochemical methods, we have shown that a new specific sequence, v-lil, is associated with a given stock of B77 avian sarcoma virus (clone 9). We prepared a DNA complementary to v-lil sequences, using substractive hybridizations, and investigated the properties of this sequence. v-lil has a genetic complexity of ca. 2,000 nucleotides and is not present in various stocks of avian sarcoma virus, avian leukosis virus, or defective leukemia virus. v-lil is not associated with B77 avian sarcoma virus isolated from the original tumor and thus has been acquired by in vitro passage of the virus on chicken embryo fibroblasts. A search for the origin of the v-lil sequence among the DNAs of different avian species has shown that a similar sequence, c-lil, is present in normal chicken DNA (1 to 2 copies per haploid genome). c-lil is not highly conserved but is present in the DNA of all chickens from the genus Gallus. The c-lil sequence is transcribed at a low level (1 to 3 copies per cell) in normal chicken embryo fibroblasts. The biological function, if any, of v-lil or its cellular equivalent has yet to be determined. PMID- 6292472 TI - Molecular cloning of a highly leukemogenic, ecotropic retrovirus from an AKR mouse. AB - SL3-3 is a leukemogenic, ecotropic retrovirus produced by a T-cell line derived from a spontaneous lymphoma of an AKR mouse. We have isolated a molecular clone of its DNA provirus from infected NIH 3T3 fibroblasts. Cloned proviral DNA produced infectious virus upon transfection onto NIH 3T3 cells. Virus derived by transfection induced lymphomas at high frequency in AKR/J, C3H(f)/Bi, CBA/J, and NFS/N mice. Heteroduplex and RNase T1 fingerprinting analyses showed that the genomes of SL3-3 and the non-leukemogenic virus, Akv, contain no major substitutions relative to one another and differ by only a few base changes. These results unambiguously show that SL3-3 is a highly leukemogenic virus and that major rearrangements of the genome relative to Akv are not required for virulence. PMID- 6292473 TI - Deletion of the nontransforming Epstein-Barr virus strain P3HR-1 causes fusion of the large internal repeat to the DSL region. AB - The nontransforming Epstein-Barr virus (EBV) strain P3HR-1 is known to have a deletion of sequences of the long unique region adjacent to the large internal repeats. The deleted region is believed to be required for initiation of transformation. To establish a more detailed map of the deletion in P3HR-1 virus, SalI-A of the transforming strain M-ABA and of P3HR-1 virus was cloned into the cosmid vector pHC79 and multiplied in Escherichia coli. The cleavage sites for BamHI, BglII, EcoRI, PstI, SacI, SacII, and XhoI were determined in the recombinant plasmid clones. Analysis of the boundary between large internal repeats and the long unique region showed that in M-ABA (EBV) the transition is different from that in B95-8 virus. The map established for SalI-A of P3HR-1 virus revealed that, in contrast to previous reports, the deletion has a size of 6.5 kilobase pairs. It involves the junction between large internal repeats and the long unique region and includes more than half of the rightmost large internal repeat. The site of the deletion in the long unique region is located between a SacI and a SacII site, about 200 base pairs apart from each other. The sequences neighboring the deletion in the long unique region showed homology to the nonrepeated sequences of the DS(R) (duplicated sequence, right) region. Sequences of the large internal repeat are thus fused to sequences of the DS(L) (duplicated sequence, left) region in P3HR-1 virus DNA under elimination of the DS(L) repeats. Jijoye, the parental Burkitt lymphoma cell line from which the P3HR-1 line is derived by single-cell cloning, is known to produce a transforming virus. Analysis of the Jijoye (EBV) genome with cloned M-ABA (EBV) probes specific for the sequences missing in P3HR-1 virus revealed that the sequences of M-ABA (EBV) BamHI-H2 are not represented in Jijoye (EBV). In Jijoye (EBV) the complete DS(L) region including the DS(L) repeats is, however, conserved. Further analysis of Jijoye (EBV) and of Jijoye virustransformed cell lines will be helpful to narrow down the region required for transformation. PMID- 6292474 TI - Antibodies against a synthetic peptide of the poliovirus replicase protein: reaction with native, virus-encoded proteins and inhibition of virus-specific polymerase activities in vitro. AB - A carboxy-terminal peptide of the poliovirus replicase protein (p63) was chemically synthesized, coupled to bovine serum albumin carrier, and injected into rabbits. The resulting antisera reacted with six virus-specific proteins from HeLa cells infected with poliovirus: NCVP 0b, NCVP 1b, NCVP 2, a protein of about 60,000 daltons, p63, and NCVP 6b. The identity of the 60,000-dalton protein is not known, but the other results were consistent with previous experimental approaches which demonstrated that p63 and the other four polypeptides have common coding sequences. An amino-terminal peptide of p63 failed to elicit an immune response in rabbits. Antibodies raised against the p63 carboxy-terminal peptide inhibited poliovirus replicase and polyuridylic acid polymerase activities in vitro, providing strong support for earlier suggestions that these activities are a property of a single virus-specific polypeptide. PMID- 6292475 TI - Epstein-Barr virus DNA XII. A variable region of the Epstein-Barr virus genome is included in the P3HR-1 deletion. AB - The P3HR-1 subclone of Jijoye differs from Jijoye and from other Epstein-Barr virus (EBV)-infected cell lines in that the virus produced by P3HR-1 cultures lacks the ability to growth-transform normal B lymphocytes (Heston et al., Nature (London) 295:160-163, 1982; Miller et al., J. Virol. 18:1071-1080, 1976; Miller et al., Proc. Natl. Acad. Sci. U.S.A. 71:4006-4010, 1974; Ragona et al., Virology 101:553-557, 1980). The P3HR-1 virus was known to be deleted for a region which encodes RNA in latently infected, growth-transformed cells (Bornkamm et al., J. Virol. 35:603-618, 1980; Heller et al., J. Virol. 38:632-648, 1981; King et al., J. Virol. 36:506-518, 1980; Raab-Traub et al., J. Virol. 27:388-398, 1978; van Santen et al., Proc. Natl. Acad. Sci. U.S.A. 78:1930-1934, 1980). This deletion is now more precisely defined. The P3HR-1 genome contains less than 170 base pairs (and possibly none) of the 3,300-base pair U2 region of EBV DNA and is also lacking IR2 (a 123-base pair repeat which is the right boundary of U2). A surprising finding is that EBV isolates vary in part of the U2 region. Two transforming EB viruses, AG876 and Jijoye, are deleted for part of the U2 region including most or all of a fragment, HinfI-c, which encodes part of one of the three more abundant cytoplasmic polyadenylated RNAs of growth-transformed cells (King et al., J. Virol. 36:506-518, 1980; King et al., J. Virol. 38:649-660, 1981; van Santen et al., Proc. Natl. Acad. Sci. U.S.A. 78:1930-1934). PMID- 6292476 TI - Hybrid selection of small RNAs by using simian virus 40 DNA: evidence that the simian virus 40-associated small RNA is synthesized by specific cleavage from large viral transcripts. AB - The simian virus 40 (SV40)-associated small RNA (SAS-RNA), approximately 64 nucleotides, is virally encoded within a region of the viral late (+) DNA strand which encodes no known protein. The SAS-RNA arises in abundance late in SV40 lytic infection. Previous data indicate that the synthesis of the SAS-RNA may be under the control of the normal late viral promoter; i.e., inhibition of transcription from the late promoter results in cessation of SAS-RNA synthesis. The synthesis of SAS-RNA was examined to determine whether the SAS-RNA is the product of cleavage from noncoding regions of nuclear late transcripts or an independent transcription product like 5S RNA, or the adenovirus VA-RNAs. The data described below suggest that SAS-RNA is cleaved from large late transcripts. In vitro transcription of DNA fragments containing the SAS-RNA coding region yielded no SAS-RNA synthesis; this result was supported by DNA sequence analysis, which indicated no promoter-like regions either within or flanking the SAS-RNA coding region. In support of a cleavage mechanism, the SAS-RNA has a 3'-phosphate end, an occurrence which is indicative of nuclease cleavage. In addition, 5'-end labeling of the SAS-RNA was possible only after calf alkaline phosphatase treatment; this indicates that the SAS-RNA is not capped. Hybrid selection analysis was used to demonstrate that separation of the SAS-RNA coding region from the normal late promoter resulted in elimination of SAS-RNA synthesis. This was demonstrated in SV40-transformed cells in which integration of a single copy of SV40 breaks the continuity of the late coding region, so that the SAS-RNA coding region is physically separated from the normal late promoter. The lack of SAS-RNA synthesis indicates that the SAS-RNA coding region cannot function as a primary transcription unit. The same result and conclusion were obtained by using a permissive cell line transformed by SV40 (COS-1 cells); here it was found that the integrated SAS-RNA coding region was not expressed even during a viable lytic infection in which the SAS-RNA could be expressed from the infecting viral genomes. The simplest conclusion drawn from the data is that the SAS-RNA is cleaved from larger late transcripts which initiate at the normal late promoter. This conclusion suggests that many of the small RNAs found in normal eucaryotic cells may be synthesized by specific cleavage rather than by primary transcription. In the course of these studies several small cellular RNAs were detected, due to their specific hybrid selection, by using SV40 DNA. Primary mapping and characterization data of these RNAs are also presented. PMID- 6292477 TI - DNA sequence of the viral and cellular src gene of chickens. 1. Complete nucleotide sequence of an EcoRI fragment of recovered avian sarcoma virus which codes for gp37 and pp60src. AB - Recovered avian sarcoma virus is a class of virus obtained from chicken tumors induced by mutants of Rous sarcoma virus which have a deletion in the src gene. We have determined the entire nucleotide sequence of a 3.1-kilobase EcoRI DNA fragment of molecularly cloned recovered avian sarcoma virus DNA. This DNA fragment contains part of the env gene and the entire src gene. Amino acid sequences of both gene products were deduced from the DNA sequences; the predicted amino acid sequences were verified by protein studies. An env protein (gp37) was found to be composed of 205 amino acids with three glycosylation sites. gp37 had a long stretch of hydrophobic residues near the carboxyl terminus. The src gene product, pp60src, was composed of 526 amino acids and contained the possible sites for tyrosine and serine phosphorylation. The amino acid sequences predicted in this study differ significantly from the amino acid sequence predicted previously for the Schmidt-Ruppin strain of Rous sarcoma virus. PMID- 6292478 TI - Effect of simian virus 40 on the temporal and spatial organization of DNA replication in Muntjac cells. AB - We examined the effects of simian virus 40 infection on the temporal and spatial organization of initiation sites for DNA replication in Muntjac cells by means of light microscopic DNA fiber autoradiography. Initiation at multiple sites along the DNA fiber in virus-infected confluent Muntjac cells was more nearly synchronous than in serum-deprived controls, although temporal control in the infected cells did not reach the level observed in cells incubated in serum enriched medium. Initiation sites in virus-infected cells appeared to be spatially closer together than in either uninfected serum-deprived or uninfected serum-enriched cells. This change did not appear to be the result of the induction or repression by simian virus 40 of clusters of replication units with new and different organizations. PMID- 6292479 TI - Simian virus 40 large T antigen is phosphorylated at multiple sites clustered in two separate regions. AB - The phosphorylation sites of simian virus 40 large T antigen were determined within the primary structure of the molecule. Exhaustive digestion of (32)P labeled large T antigen with trypsin generated six major phosphopeptides which could be separated in a newly developed isobutyric acid-containing chromatography system. By partial tryptic digestion, large T antigen was cleaved into an amino terminal fragment of 17,000 daltons and overlapping fragments from the carboxy terminal region ranging in size between 71,000 and 13,000 daltons. The location of the phosphopeptides was then determined by fingerprint analyses of individual fragments. Their physical properties were analyzed by sizing on polyacrylamide gels and by sequential digestion and peptide mapping; their amino acid composition was determined by differential labeling with various amino acids. The amino-terminal 17,000-dalton fragment gave rise to only one phosphopeptide (phosphopeptide 3) that contained half of the phosphate label incorporated into large T antigen. It contained phosphoserine and phosphothreonine sites, all of which were clustered within a small segment between Cys(105) and Lys(127). This segment contained five serines and two threonines. Among these, Ser(106), Ser(123), and Thr(124) were identified as phosphorylated residues; in addition, either one or both of Ser(111) and Ser(112) were phosphorylated. The neighboring residues, Ser(123) and Thr(124), were found in three different phosphorylation states in that either Ser(123) or Thr(124) or both were phosphorylated. Phosphopeptides 1, 2, 4, 5, and 6 were all derived from a single fragment extending 26,000 daltons upstream from the carboxy terminus of large T antigen. Phosphopeptide 6 was identical with the previously determined phosphothreonine peptide phosphorylated at Thr(701). Phosphopeptides 1, 2, 4, and 5 contained only serine-bound phosphate. Phosphopeptides 1, 2, and 4 represented overlapping peptides, all of which were phosphorylated at Ser(639) located next to a cluster of six acidic residues. In phosphopeptide 5, a large peptide ranging from Asn(653) to Arg(691), at least two of seven serines were phosphorylated. Thus, large T antigen contains at least eight phosphorylation sites. Their clustering within two separate regions might correlate with structural and functional domains of this protein. PMID- 6292481 TI - Recombination occurs mainly between parental genomes and precedes DNA replication in pseudorabies virus-infected cells. AB - The experiments described in this paper were part of an attempt to determine the mechanisms involved in the isomerization of the pseudorabies virus genome. To this end, [(14)C]thymidine-labeled parental virus DNA that was transferred to progeny virions produced by cells incubated in medium containing bromodeoxy uridine was analyzed in neutral and alkaline CsCl density gradients. The buoyant density of the (14)C-labeled DNA indicated that the parental DNA strands had retained their integrity and had not undergone breakage and reunion with progeny DNA strands; neither massive intermolecular nor intramolecular recombination had occurred after replication of the DNA. Whereas breakage and reunion between parental and progeny virus DNA strands were not detectable, these processes were observed between differentially density-labeled parental DNAs. Furthermore, the frequency of recombination between progeny DNAs accumulating in the cells was low. These results indicate that in pseudorabies virus-infected rabbit kidney cells recombination occurs mainly between parental genomes and precedes DNA replication. An analysis of the kinetics of appearance of recombinants between pairwise combinations of temperature-sensitive mutants also indicated that recombination is an early event. The ratio between the number of recombinant virions and the number of temperature-sensitive mutant virions produced by the cells remained the same throughout infection. Since the relative amounts of viral DNAs synthesized early and late during the infective process that were integrated into virions were approximately the same, it appears that late viral DNA did not experience an increased number of recombinational events compared with early viral DNA. These results, which reinforce the conclusion reached from the results of the analysis of the behavior of the parental DNA molecules in density shift experiments, indicate that recombination is an early event. PMID- 6292480 TI - DNA sequence of the viral and cellular src gene of chickens. II. Comparison of the src genes of two strains of avian sarcoma virus and of the cellular homolog. AB - The nucleotide sequence of the src gene and flanking regions of the Schmidt Ruppin strain of Rous sarcoma virus (SR-A) was determined. The src region of SR-A was very homologous to that of recovered avian sarcoma virus (rASV1441), with only 17 differences among 1,578 nucleotides. The size of the predicted protein was 526 amino acids in both viruses, of which 6 amino acids were different. The differences in nucleotides and amino acids between the two viruses localized within the 5' two-thirds of the src coding region. There were also viruses localized within the 5' two-thirds of the src coding region. There were also some differences in the region flanking the 5' end of src. Since rASVs are considered to be recombinatns between deletion mutants of SR-A and cellular-src (c-src) sequences, several segments of c-src DNA were also sequenced to understand the molecular basis for the recombination. At 14 of 17 bases where SR-A and rASV1441 differed, rASV1441 had the same sequence as c-src. Three of these sequences corresponded to sequences of oligonucleotides which were previously identified in RNAs of nearly all isolates of rASV but which were absent in SR-A RNA. In the 5' flanking sequences of the src gene, c-src was more similar to rASV1441 than to SR A. These results confirm the cellular origin of the src sequences of rASVs and provide information about the possible sites of the recombination. PMID- 6292482 TI - Molecular analysis of the c-myc locus in normal tissue and in avian leukosis virus-induced lymphomas. AB - We isolated molecular clones of the provirus-host cell junctions (tumor junction fragments) from two avian leukosis virus-induced lymphomas and compared the structures of these clones with a clone of the normal c-myc gene. Restriction mapping and DNA sequencing demonstrated that normal proviral integration events occurred adjacent to c-myc in both tumors, without gross structural alteration of c-myc. The right long terminal repeat of an avian leukosis virus provirus is integrated upstream from the bulk of the c-myc coding sequences and oriented such that transcription can initiate within the long terminal repeat and proceed downstream into c-myc. A comparison of a tumor junction fragment with the v-myc gene showed that there are two regions of v-myc-related sequences (which are probably exons) separated by 1 kilobase of sequences unrelated to v-myc (probably an intron). A DNA sequence analysis of the tumor junction fragments suggested that integration had occurred in exons adjacent to splice donor sites. This suggests that there are additional exons and introns in c-myc. Based on these findings, a model is proposed for the genesis of the tumor-specific RNAs containing viral-5' and c-myc information in avian leukosis virus-induced lymphomas. PMID- 6292483 TI - Novel phenotype of RNA synthesis expressed by vesicular stomatitis virus isolated from persistent infection. AB - Vesicular stomatitis virus (VSV) stocks isolated from two persistently infected mouse L-cell lines (designated VSV-PI stocks) express an altered phenotype of RNA synthesis. This phenotype is different from the RNA synthesis phenotype expressed by the viruses used to initiate the persistently infected lines, wild-type VSV and VSV ts-0-23 (a group III, ts-, RNA+ mutant). At 34 and 37 degrees C in L cells productively infected with VSV-PI stocks derived from the two cell lines, transcription of virus mRNA was significantly reduced, whereas replication of the 40S genomic RNA species was enhanced compared with wild-type VSV or ts-0-23. At 34 and 37 degrees C, both VSV-PI stocks replicated with equal or greater efficiency than wild-type VSV; 37 degrees C was the temperature at which the persistently infected cultures were maintained. At 40 degrees C, both VSV-PI stocks were temperature sensitive, and clonal VSV-PI isolates from both cell lines belong to complementation group I (RNA-). Standard ts- mutants (derived by mutagenesis of wild-type VSV) belonging to RNA- complementation groups I, II, and IV do not express the VSV-PI RNA synthesis phenotype at the permissive temperature, making this phenotype distinctive to persistent infection. Since the two VSV-PI populations from persistently infected cell lines initiated with different viruses both evolved this unique phenotype of RNA synthesis, the expression of this phenotype may play an important role in the maintenance of persistence. PMID- 6292484 TI - Polyoma virus early and late mRNAs in productively infected mouse 3T6 cells. AB - We mapped polyoma virus-specific mRNAs isolated from productively infected mouse 3T6 cells on the viral genome by analyzing nuclease S1-resistant RNA-DNA hybrids. The polyoma early mRNAs, which code for the three T antigens, have several 5' ends near 73 map units (m.u.). During the late phase of infection an additional 5' end is found near 71 m.u. All of the major early mRNAs have common 3' ends at 26.01 m.u. There is a minor species of early mRNA with a 3' end at 99.05 m.u. There are two proximal and two distal splice junctions in the early region which are used to generate three different spliced early mRNAs. There are three late mRNAs encoding the three virion proteins, VP1, VP2, and VP3. The late mRNAs have common 3' ends at 25.34 m.u. The late mRNAs have heterogeneous 5' leader sequences derived from the region between 65.53 and 68.42 m.u. The leader sequences are joined to the bodies of the messages coding for VP2, VP3, and VP1 at 66.59, 59.62, and 48.57 m.u., respectively. These results confirm and extend previous analyses of the fine structure of polyoma mRNAs. PMID- 6292485 TI - Characterization of an infective molecular clone of the B-tropic, ecotropic BL/Ka(B) murine retrovirus genome. AB - Using molecular cloning techniques, we amplified the unintegrated, linear proviral DNA of the BL/Ka(B) virus, a non-leukemogenic retrovirus of mouse strain C57BL/Ka. Two independent clones in lambda phage vector 607 and one subclone in pBR322 were infective when transfected into mouse fibroblasts. Analysis of the progeny virus revealed biological properties and a restriction map identical to those of the parental viral shock. Comparison of the restriction map with the maps of other ecotropic murine viruses reveals many similarities. Particularly interesting is the comparison of the N-tropic Akv virus and the B-tropic BL/Ka(B) virus. The long terminal repeats of the two viruses are virtually identical, as are 22 of 23 restriction sites located outside of the region which spans from 1.8 to 3.8 kilobases from the left end of the genome. Within this region, however, only three of nine sites examined are shared. This suggests that the BL/Ka(B) virus was derived from an endogenous N-tropic virus closely related to Akv by recombinational events which altered the sequence in the last half of the gag gene and the first third of the pol gene. This change is probably responsible for the observed difference in the Fv-1 tropism of the two viruses. PMID- 6292486 TI - Adenovirus type 2 fiber mRNA synthesis: no evidence for a cytoplasmic processing pathway. AB - Adenovirus type 2 fiber mRNA exists in several forms in the cytoplasm which differ in the presence or absence of extra 5'-leader segments (L. T. Chow and T. R. Broker, Cell 15:497-510, 1978). We have investigated the possibility that forms possessing extra leader segments serve as precursors to the mature form in the cytoplasm. Pulse-labeled fiber mRNA became considerably shorter (150 to 250 bases) during a chase; however, most of the pulse-labeled species failed to hybridize to DNA fragments known to encode extra leader segments. Moreover, the entire decrease in size appeared to be due to extensive shortening of the polyadenylic acid tail. Mature-sized fiber mRNA was synthesized normally in cells infected with the nondefective adenovirus type 2-simian virus 40 hybrid virus Ad2+ND5, in which the region encoding the extra leader segments is deleted. These results indicate that the additional 5'-leader segments present in wild-type adenovirus type 2 fiber mRNA are not required for the production of mature fiber mRNA and that species that possess them are not cytoplasmic precursors to the mature form. PMID- 6292487 TI - Endo-beta-N-acetylglucosaminidase H sensitivity of precursors to herpes simplex virus type 1 glycoproteins gB and gC. AB - The endoglycosidase endo-beta-N-acetylglucominidase H (endo H) was used to examine the nature of the oligosaccharides associated with the herpes simplex virus type 1 glycoproteins gA, gB, and gC. Immunoprecipitates from detergent extracts of infected cells, using monospecific antisera to gAB and gC, were treated with endo H. The low-molecular-weight precursor to gC, pgC(105), was found to be sensitive to endo H. Removal of the endo H-sensitive oligosaccharide chains from pgC(105) resulted in a protein with an apparent molecular weight of 75,000. In contrast, the fully glycosylated gC was not sensitive to endo H treatment. These results suggested that the oligosaccharide chains of pgC(105) were primarily of the simple high-mannose type. Both gA and gB were sensitive to endo H treatment; however, gB appeared to be only partially susceptible, whereas [3H]mannose-labeled gA was not detectable after endo H treatment. These results that gB contained both complex- and simple-type oligosaccharides, and gA contained only simple-type oligosaccharides. An accumulation of the high-mannose glycoproteins pgC(105) and gA was observed in monensin-treated infected cells with a concomitant inhibition of gB and gC. Glycoproteins gA and pgC(105) synthesized in the presence of monensin were also sensitive to endo H treatment. PMID- 6292488 TI - Assessment of the base sequence homology between the two subtypes of equine herpesvirus 1. AB - The magnitude of the genetic relatedness of the two antigenic subtypes of equine herpesvirus 1 (EHV-1) was determined by DNA-DNA reassociation kinetics. Denatured, labeled viral DNA from one EHV-1 subtype was allowed to reassociate in the presence or absence of the unlabeled heterologous viral DNA. The initial rate of reassociation of either labeled viral DNA was increased by the presence of the heterologous viral DNA to an extent indicating 10 to 20% homology between the two EHV-1 genomes. Similar estimates of the amount of homology between the genomes of the two EHV-1 subtypes were obtained by determining the maximum fraction of labeled viral DNA that could be made resistant to S1 nuclease by hybridization with a large molar excess of the unlabeled, heterologous viral DNA. Analysis of the thermal stability of the subtype 1-subtype 2 heteroduplex DNA indicated approximately 30% base pair mismatching within the hybrid DNA molecules. Cross hybridization of 32P-labeled virion DNA to nitrocellulose blots of restriction endonuclease cleavage fragments of each EHV-1 subtype DNA indicated that the observed homology between the two viruses was nonuniformly distributed with the viral genome. No homology could be detected between the DNA of either EHV-1 subtype and that of a strain of equine cytomegalovirus (EHV-2). The data suggest that the two biotypes of EHV-1 have arisen by divergent evolution from a common progenitor herpesvirus. PMID- 6292489 TI - Two colinear and spliced viral transcripts are present in non-virus-producing benign and malignant neoplasms induced by the shope (rabbit) papilloma virus. AB - The nature of Shope virus-specific RNA was investigated in non-virus-producing Shope (rabbit) papilloma virus-induced benign and malignant domestic rabbit tumors and in a cell line derived from the VX-7 transplantable carcinoma. RNA transfer (Northern) blot analysis of polyadenylated RNA isolated from whole cell extracts of all three sources was resolved into two major bands of 1.3 and 2.0 kilobases. Two additional minor bands of 3.5 and 4.8 kilobases could be seen in some analyses. RNA of the VX-7 cell line was further separated into nuclear, cytoplasmic, and polysomal fractions. The cytoplasmic fraction only contained the 1.3- and 2.0-kilobase species, and virus-specific RNA was found to be associated with polysomes. The two major transcripts present in VX-7 cells were mapped by hybridization of RNA transfer blots with subgenomic probes, and the results indicated that both transcripts are spliced and are most likely colinear. Our results are consistent with the suggestion that the two viral transcripts are necessary for induction and maintenance of the neoplasms. PMID- 6292490 TI - Molecular cloning of circular unintegrated DNA of two types of the SEATO strain of gibbon ape leukemia virus. AB - Closed circular unintegrated DNA of the SEATO strain of gibbon ape leukemia virus (GaLV-S) was isolated from canine thymus fibroblasts after cocultivation with chronically infected bat lung fibroblasts. Restriction endonuclease HindIII cleaves GaLV-S DNA once, thus allowing isolation and cloning of HindIII-digested unintegrated DNA in a permitted form. Two clones isolated in the vector, Charon 21A, were nearly identical by restriction enzyme mapping to each of the two types of GaLV-S previously observed. These two types differ at a single SalI site. Unlike previous maps of GaLV-S proviral DNA, however, both clones lack SstI sites in the long-terminal-repeat units. Both the GaLV-S clones and the major species of GaLV-S proviral DNA contain an EcoRI site in the long-terminal-repeat units. The presence of this EcoRI site and the absence of an SstI site in the GaLV-S long-terminal-repeat units differentiate it from all other known GaLV strains and from the closely related nononcogenic simian sarcoma-associated virus. Heteroduplex comparisons of each of the two clones to clones of simian sarcoma associated virus show no obvious deletion or substitution loops. This suggests that the ability of GaLV-S to induce myeloid leukemia in gibbon apes in not due to an acquired onc gene. PMID- 6292491 TI - Long internal direct repeat in Epstein-Barr virus DNA. AB - The nucleotide sequence of the long internal reiteration. IR1, of Epstein-Barr Virus DNA has been determined. The repeat unit is 3,071 base pairs which are 66.8% guanine plus cytosine. There is a CCAAT sequence 39 nucleotides 5' to a TATAA, which could indicate a promotor for transcription. The longest open reading frame is 1,124 base pairs. Also within IR1 is a sequence homologous to the papovavirus origin of DNA replication. The ori-like sequence is within a long palindromic region which is 500 base pairs overall. The palidromic region shares common features with the Alu family members and with eucaryotic transposable elements. The juncture between the short unique region (U1) and IR1 is also sequenced. The transition occurs in BamHI-C at 1,214 base pairs before the BamHI site in the first repeat of IR1. The transition from IR1 to the rightward unique region (U2) has been reported to be at 636 base pairs after the BamHI site in the last repeat of IR1. Thus, relative to the start of IR1 at the juncture with U1, the last copy of IR1 is a partial repeat which contains only the beginning 1,850 base pairs. PMID- 6292492 TI - Structure of nonintegrated, circular Herpesvirus saimiri and Herpesvirus ateles genomes in tumor cell lines and in vitro-transformed cells. AB - Nonintegrated, circular DNA molecules of Herpesvirus saimiri and Herpesvirus ateles were found in five lymphoid cell lines originating from tumor tissues or established by in vitro immortalization of T lymphocytes. The arrangement of unique (L) and repetitive (H) DNA sequences in circular viral genomes was analyzed by partial denaturation mapping followed by visualization with an electron microscope. Three types of circular viral DNA structures were found. (i) The virus-producing cell line RLC, which is derived from an H. ateles-induced rabbit lymphoma, contains circular viral genomes which consist of a single L-DNA and a single H-DNA region, both the same length as in virion DNA. (ii) The circular viral genomes of the nonproducer cell lines H1591 and A1601, in vitro transformed by H. saimiri and H. ateles, respectively, have deletions in the unique L-DNA region and larger H-DNA regions. Cell line A1601 lacks about 8% of virion L-DNA, and H1591 cells lack about 40% of viral L-DNA information. (iii) The nonproducing H. saimiri tumor cell lines 1670 and 70N2 harbor viral genomes with two L-DNA and two H-DNA regions, respectively. Both types of circular molecules have a long and a short L-segment. The sequence arrangements of circular DNA molecules from H. saimiri-transformed cell lines were compared with those of linear virion DNA by computer alignment of partial denaturation histograms. The L-DNA deletion in cell line H1591 was found to map in the right half of the virion DNA. Comparison of the denaturation patterns of both L regions of cell lines 1670 and 70N2 identified the short L regions as subsets of the long L regions. Thus, circular viral DNA molecules of all four nonproducer cell lines represent defective genomes. PMID- 6292493 TI - Simple repeat sequence in Epstein-Barr virus DNA is transcribed in latent and productive infections. AB - The BamHI K region of Epstein-Barr virus DNA is transcribed in latently infected cells from Burkitt tumors and in growth-transformed B-lymphocytes latently infected with Epstein-Barr virus. We determined the nucleotide sequence of a 1,153-base pair HinfI fragment in BamHI fragment K from the B95-8 Epstein-Barr virus isolate. The fragment contains a remarkable 708-base pair simple sequence repeat array, designated IR3, which is composed of only three nucleotide triplet elements: GGG, GCA, and GGA. The triplets are organized into three repeat units: GCAGGA, GCAGGAGGA, and GGGGCAGGA. Immediately 3' of IR3 are tandem nearly perfect direct repeats of two different 24-base pair sequences. IR3 is conserved at a colinear position in the DNAs of other Epstein-Barr virus isolates, and a homologous sequence maps at the same location in the genome of a genetically related baboon herpesvirus, herpesvirus papio. IR3 is transcribed from left to right in latently infected, growth-transformed IB4 cells. It encodes part of a 2.0-kilobase exon of the 3.7-kilobase cytoplasmic polyadenylated RNA previously detected in IB4 cells (van Santen et al., Proc. Natl. Acad. Sci. U.S.A. 78:1930 1934, 1981). IR3 also encodes parts of 2.4- and 1.0-kilobase RNAs in productively infected B95-8 cells. PMID- 6292494 TI - Nucleotide sequence of bacteriophage f1 DNA. AB - The nucleotide sequence of the DNA of the filamentous coliphage f1 has been determined. In agreement with earlier conclusions, the genome was found to comprise 6,407 nucleotides, 1 less than that of the related phage fd. Phage f1 DNA differs from that of phage M13 by 52 nucleotide changes, which lead to 5 amino acid substitutions in the corresponding proteins of the two phages, and from phage fd DNA by 186 nucleotide changes (including the single-nucleotide deletion), which lead to 12 amino acid differences between the proteins of phages f1 and fd. More than one-half of the nucleotide changes in each case are found in the sequence of 1,786 nucleotides comprising gene IV and the major intergenic region between gene IV and gene II. The sequence of this intergenic region (nucleotides 5501 to 6005) of phage f1 differs from the sequence reported by others through the inclusion of additional single nucleotides in eight positions and of a run of 13 nucleotides between positions 5885 and 5897, a point of uncertainty in the earlier published sequence. The differences between the sequence of bacteriophage f1 DNA now presented and a complete sequence for the DNA previously published by others are discussed, and the f1 DNA sequence is compared with those of bacteriophages M13 and fd. PMID- 6292496 TI - Glycoproteins of herpes simplex virus type 2 as defined by monoclonal antibodies. AB - We used monoclonal antibodies reacting with glycoproteins specified by herpes simplex virus type 2 (HSV-2) to characterize the individual antigens in terms of structure, processing, and kinetics of synthesis in BHK or Vero infected cells. Our results provided a direct demonstration of the structural identity of the gA and gB proteins of HSV-2 as well as confirmation of the existence of type specific and type-common domains within the gD molecule. They also show that, with the exception of gC, processing of the viral glycoproteins differs to some extent in Vero and BHK infected cells, possibly as a result of different efficiency of glycosylation or different processing of underglycosylated and unglycosylated products in the two cell types. Finally, we showed that individual HSV-2 glycoproteins are synthesized at greatly different times during the infectious cycle, possibly in response to their different roles in virus replication and assembly. PMID- 6292495 TI - Avian retrovirus pp32 DNA-binding protein. I. Recognition of specific sequences on retrovirus DNA terminal repeats. AB - The avian retrovirus pp32 protein possesses a DNA-nicking activity which prefers supercoiled DNA as substrate. We have investigated the binding of pp32 to avian retrovirus long terminal repeat (LTR) DNA present in both supercoiled and linear forms. The cloned viral DNA was derived from unintegrated Schmidt-Ruppin A (SRA) DNA. A subclone of the viral DNA in pBR322 (termed pPvuII-DG) contains some src sequences, tandem copies of LTR sequences, and partial gag sequences in the order src-U(3) U(5):U(3) U(5)-gag. Binding of pp32 to supercoiled pPvuII-DG DNA followed by digestion of this complex with a multicut restriction enzyme (28 fragments total) permitted pp32 to preferentially retain on nitrocellulose filters two viral DNA fragments containing only LTR DNA sequences. In addition, pp32 also preferentially retained four plasmid DNA fragments containing either potential promoters or Tn3 "left-end" inverted repeat sequences. Mapping of the pp32 binding sites on viral LTR DNA was accomplished by using the DNase I footprinting technique. The pp32 protein, but not the avian retrovirus alphabeta DNA polymerase, is able to form a unique protein-DNA complex with selected regions of either SRA or Prague A LTR DNAs. Partial DNase I digestion of a 275 base pair SRA DNA fragment complexed with pp32 gives upon electrophoresis in denaturing gels a unique ladder pattern, with regions of diminished DNase I susceptibility from 6 to 10 nucleotides in length, in comparison with control digests in the absence of protein. The binding of pp32 to this fragment also yields enhanced DNase I-susceptible sites that are spaced between the areas protected from DNase I digestion. The protected region of this unique complex was a stretch of 170 +/- 10 nucleotides that encompasses the presumed viral promoter site in U(3), which is adjacent to the src region, extends through U(5), and proceeds past the joint into U(3) for about 34 base pairs. No specific protection or DNase I enhancement by pp32 was observed in experiments with a 435-base pair SRA DNA fragment derived from a part of U(3) and the adjacent src region or a 55 base pair DNA fragment derived from another part of U(3). The DNA sequence of Prague A DNA at the fused LTRs differs from that of SRA DNA. The alteration in the sequence at the juncture of the LTRs prevented pp32 from forming a stable complex in this region of the LTR. Our results are relevant to two aspects of the interaction between pp32 and LTR DNA. First, the pp32 protein in the presence of selected viral DNA restriction fragments possibly forms a higher order oligomer analogous to Escherichia coli DNA gyrase-DNA complexes or eucaryotic nucleosome structures. Second, the specificity of the binding suggests a role for pp32 and the protected DNA sequences in the retrovirus life cycle. The preferred sequences to which pp32 binds include two adjacent 15-base pair inverted terminal repeats at the joint between U(5) and U(3) in SRA DNA. This region is involved in circularization of linear DNA and is perhaps the site that directs integration into cellular DNA. PMID- 6292497 TI - Sequential synthesis of 5'-proximal vesicular stomatitis virus mRNA sequences. AB - We examined the kinetics of synthesis in vitro of the 5' ends of the vesicular stomatitis virus mRNAs by analysis of specific RNase T1 oligonucleotides located near the 5' ends of the mRNAs. Our results indicate that, like synthesis of full length mRNAs, the 5' ends of the mRNAs are synthesized sequentially, following the gene order N, NS, and M. Additional experiments with UV-irradiated virus demonstrated that synthesis of the mRNA regions containing these oligonucleotides is dependent on synthesis of the mRNA from the preceding gene. These results are inconsistent with a model of vesicular stomatitis virus transcription involving simultaneous initiation and presynthesis of leader RNAs 30 to 70 nucleotides long for each mRNA. We also characterized two small RNA species whose synthesis is highly resistant to UV irradiation. Partial sequence analysis indicates that these RNAs are a 5'-capped fragment of the N mRNA and a 5' fragment of the leader RNA. PMID- 6292498 TI - Virion DNA of ground squirrel hepatitis virus: structural analysis and molecular cloning. AB - The structure of the encapsidated DNA genome of ground squirrel hepatitis virus (GSHV) has been examined by restriction endonuclease cleavage, nucleic acid hybridization, and molecular cloning. GSHV virion DNA is a relaxed circular molecule of approximately 3,200 bases in length; most molecules harbor an extensive single-stranded region which is largely confined to one-half of the genome. The full-length viral DNA strand is covalently bound to protein. The single-stranded region can be repaired in vitro by the action of the endogenous virion polymerase, exogenously added DNA polymerase from avian myeloblastosis virus, or both. Restriction enzyme cleavage of viral DNA from different isolates demonstrated that multiple variants of GSHV exist in nature. The genomes of two such strains have been cloned in Escherichia coli, and their physical maps have been determined. Nucleic acid hybridization studies revealed that the strains share sequence homology with the DNA of human hepatitis B virus. Regions homologous to the coding regions for the surface and core antigens of human hepatitis B virus have been localized on the GSHV chromosome. Molecular cloning experiments have also led to the identification of a region of the viral genome which is altered in a procaryotic host. PMID- 6292499 TI - Biosynthesis of virus-specific proteins in cells infected with infectious bursal disease virus and their significance as structural elements for infectious virus and incomplete particles. AB - It has previously been shown that infectious bursal disease virus is a naked icosahedral particle with a diameter of about 60 nm and a genome consisting of two segments of double-stranded RNA (Muller et al., J. Virol. 31:584-589, 1979). One of the two major structural polypeptides (molecular weight, 40,000) of this virus could not be found in lysates of infected cells; it is derived from a precursor polypeptide demonstrable inside the cells in relatively large quantities and seems to be processed during virus assembly or later. The precursor molecule is regularly present in the infectious virus particle (buoyant density, 1.33 g/ml) in minor proportions, but it represents an outstanding structural element of incomplete noninfectious particles ("top components"; buoyant density, 1.29 g/ml) which contain viral RNA. This type of incomplete particles is mainly produced by chicken embryo fibroblasts in contrast to lymphoid cells from the bursa of Fabricius. Precursor-product relationships also seem to exist in the biosynthesis of the other viral polypeptides. In contrast to some other viruses with a segmented double-stranded RNA genome, none of the structural proteins of infectious bursal disease virus is appreciably glycosylated. PMID- 6292500 TI - Molecular cloning and characterization of human papilloma virus DNA derived from a laryngeal papilloma. AB - Papilloma virus DNA from a laryngeal papilloma was cloned in phage lambda L 47 and characterized after cleavage with different restriction enzymes. Hybridization with the DNAs of human papilloma virus types 1, 2, 3, 4, 5, and 8 showed no homology under stringent hybridization conditions. Human papilloma virus type 6 DNA, however, was partially identical to laryngeal papilloma virus DNA; different restriction enzyme fragments hybridizing with the other DNA were identified on each genome. The degree of homology was determined by reassociation kinetics to be 25%. According to the present nomenclature, laryngeal papilloma virus therefore represents a different type of human papilloma virus and is tentatively designated as human papilloma virus type 11. Sequences homologous to laryngeal papilloma virus DNA were also found in four of nine additional laryngeal papillomas. Attempt to detect homologous DNA in 12 carcinomas of the larynx were negative. PMID- 6292501 TI - DNA clone of avian Fujinami sarcoma virus with temperature-sensitive transforming function in mammalian cells. AB - We have molecularly cloned an integrated proviral DNA of Fujinami sarcoma virus (FSV) into a lambda phage vector and further subcloned it into plasmid pBR322. The source of provirus was a quail nonproducer cell clone transformed by FSV. The FSV strain used is temperature sensitive in the maintenance of transformation of avian cells. The recombinant plasmid was shown to contain an entire FSV genome by fingerprinting the hybrids formed with 32P-labeled FSV RNA. This analysis also revealed a previously undetected env-related sequence in FSV which represents the 3' end of the gp85 env gene. A physical map of cloned FSV DNA identifying sites of several restriction enzymes is described. Upon transfection, FSV DNA cloned in pBR322 transformed mouse NIH-3T3 cells, which proved to be temperature sensitive in maintaining transformation. Phosphorylation but not synthesis of p140, the only known gene product of FSV, was also temperature sensitive in these cells. The correlation between transformation and phosphorylation of p140 suggests that phosphorylation of p140 is necessary for transformation of mouse cells, as was shown previously for avian cells. These results provide direct genetic evidence that the mechanisms for maintaining transformation of mammalian and avian cells involve the same FSV gene product, p140. Homology was detected by hybridization between transformation-specific sequences of FSV DNA and certain restriction endonuclease-resistant fragments of cellular DNA of two avian species, chicken and quail. Under the same conditions homology was also detected with DNA of non avian species, although apparently to a lower degree than with avian cells. PMID- 6292502 TI - Avian myeloblastosis provirus cloned in a lambda bacteriophage is leukemogenic. AB - The avian myeloblastosis virus provirus inserted in a lambda bacteriophage, recombinant clone 11A1-1 (Souza et al., Proc. Natl. Acad. Sci. U.S.A. 77:3004 3008, 1980), was transfected into chicken embryo fibroblasts which had been preinfected with either Rous-associated virus type 61 or the transformation defective avian sarcoma virus tdB77. Within 4 to 5 h after transfection, the cells were injected into 16-day-old chicken embryos or 1-day-old chicks. Acute myeloblastic leukemia developed after a long latent period. Filtered (0.22 micrometer pores) supernatant of transformed buffy-coat cell cultures from one leukemic chicken of the lambda 11A1-1 (tdB77) group rapidly transformed yolk sac cells in vitro. Results from an infectivity interference assay and analysis of proviral DNA fragments generated with restriction endonucleases were consistent with the presence in leukemic cells of defective avian myeloblastosis virus and tdB77 as the helper virus. PMID- 6292503 TI - Single-stranded regions on unintegrated avian retrovirus DNA. AB - Using chromatography on benzoylated naphthoylated DEAE-cellulose, we found that greater than 99.5% of the unintegrated linear viral DNA species detected in quail embryo cells infected with Rous sarcoma virus contained single-stranded regions, even at 16 h after infection. These regions were distributed across the genome and, on average, were primarily of plus-strand DNA. Within most of the linear viral DNA species, the minus strand was interpreted as being of genome size with two copies of the large terminal redundancy, LTR. In contrast, the plus strands in the linear viral DNA species were exclusively subgenomic. PMID- 6292504 TI - Structure and biochemical functions of four simian virus 40 truncated large-T antigens. AB - The structure of four abnormal T antigens which are present in different simian virus 40 (SV40)-transformed mouse cell lines was studied by tryptic peptide mapping, partial proteolysis fingerprinting, immunoprecipitation with monoclonal antibodies, and in vitro translation. The results obtained allowed us to deduce that these proteins, which have apparent molecular weights of 15,000, 22,000, 33,000 and 45,000, are truncated forms of large-T antigen extending to different amounts into the amino acid sequences unique to large-T. The proteins are all phosphorylated, probably at a site between amino acids 106 and 123. The mRNAs coding for the proteins probably contain the normal large-T splice but are shorter than the normal transcripts of the SV40 early region. The truncated large Ts were tested for the ability to bind to double-stranded DNA-cellulose. This showed that the 33,000- and 45,000-molecular-weight polypeptides contained sequences sufficient for binding under the conditions used, whereas the 15,000- and 22,000-molecular-weight forms did not. Together with published data, this allows the tentative mapping of a region of SV40 large-T between amino acids 109 and 272 that is necessary and may be sufficient for the binding to double stranded DNA-cellulose in vitro. None of the truncated large-T species formed a stable complex with the host cell protein referred to as nonviral T-antigen or p53, suggesting that the carboxy-terminal sequences of large-T are necessary for complex formation. PMID- 6292505 TI - Structural and biological analysis of integrated polyoma virus DNA and its adjacent host sequences cloned from transformed rat cells. AB - EcoRI fragments containing integrated viral and adjacent host sequences were cloned from two polyoma virus-transformed cell lines (7axT and 7axB) which each contain a single insert of polyoma virus DNA. Cloned DNA fragments which contained a complete coding capacity for the polyoma virus middle and small T antigens were capable of transforming rat cells in vitro. Analysis of the flanking sequences indicated that rat DNA had been reorganized or deleted at the sites of polyoma virus integration, but none of the hallmarks of retroviral integration, such as the duplication of host DNA, were apparent. There was no obvious similarity of DNA sequences in the four virus-host joins. In one case the virus-host junction sequence predicted the virus-host fusion protein which was detected in the transformed cell line. DNA homologous to the flanking sequences of three out of four of the joins was present in single copy in untransformed cells. One copy of the flanking host sequences existed in an unaltered form in the two transformed cell lines, indicating that a haploid copy of the viral transforming sequences is sufficient to maintain transformation. The flanking sequences from one cell line were further used as a probe to isolate a target site (unoccupied site) for polyoma virus integration from uninfected cellular DNA. The restriction map of this DNA was in agreement with that of the flanking sequences, but the sequence of the unoccupied site indicated that viral integration did not involve a simple recombination event between viral and cellular sequences. Instead, sequence rearrangements or alterations occurred immediately adjacent to the viral insert, possibly as a consequence of the integration of viral DNA. PMID- 6292506 TI - Phosphorylation and dephosphorylation alter the structure of D2 hybrid T antigen. AB - D2 hybrid T antigen is a protein closely related to simian virus 40 large T antigen and is synthesized in large quantities in cells infected with Ad2+D2, an adenovirus-simian virus 40 hybrid. We have analyzed the effects of phosphorylation on the structure and DNA binding of this protein. On nondenaturing pore-gradient gels, the purified protein migrated with an apparent molecular weight of 135,000, with a minor band at 330,000 molecular weight. In vitro phosphorylation catalyzed by the protein kinase activity associated with the protein resulted in a structural change so that most of it migrated with an apparent molecular weight of 740,000. Treatment of the phosphorylated form of the protein with alkaline phosphatase (which removed 95% of the phosphate) caused the disappearance of the 740,000-molecular-weight form and reappearance of the smaller forms. Partial tryptic digestion showed that D2 T antigen has two major regions of phosphorylation, only one of which was phosphorylated in vitro. The region phosphorylated in vitro was responsible for the aggregation of D2 T antigen and was tentatively assigned to the N-terminal part of the protein. As shown by protein blotting onto nitrocellulose filters, it was mainly the form of 740,000 molecular weight that bound to simian virus 40 DNA. However, sucrose gradient analyses showed that only a fraction of the in vitro-phosphorylated protein bound to DNA, suggesting that aggregation alone is not sufficient for binding. PMID- 6292507 TI - Herpes simplex virus glycoprotein gA/B: evidence that the infected Vero cell products comap and arise by proteolysis. AB - We recently reported (Pereira et al., Proc. Natl. Acad. Sci. U.S.A. 78:5202-5206, 1981) that herpes simplex virus 1 and 2 glycoproteins, previously designated gA and gB, could not be differentiated by a bank of independently derived type specific and type-common monoclonal antibodies. We also reported that from lysates of infected Vero cells, all but one monoclonal antibody precipitated gA/B glycoproteins which had faster electrophoretic mobility than the corresponding infected HEp-2 cell glycoproteins and a set of three small polypeptides which we designated g(A + B) reactive polypeptides 1, 2, and 3. Antibody H368, the single exception, failed to react with the gA/B glycoproteins or related antigens accumulating in infected Vero cells. In this paper, we report the following results. (i) The high-apparent-molecular-weight gA/B glycoproteins accumulating in infected HEp-2 cells were cleaved by a proteolytic enzyme contained in Vero cell lysates to yield more rapidly migrating proteins that were indistinguishable from authentic Vero cell gA/B glycoproteins. Like its authentic counterpart, the cleaved gA/B glycoproteins failed to react with H368 monocolonal antibody. In addition, the lysate cleaved HEp-2 cell gA/B glycoproteins into g(A + B) reactive polypeptides 2 and 3. (ii) The proteolytic activity contained in the uninfected cell lysates was inhibited by N-alpha-p-tosyl-l-lysine chloromethyl ketone and is therefore trypsin-like. (iii) Pulse-chase experiments indicated that the cleavage of gA/B glycoproteins occurred during or soon after translation but that the accumulation of g(A + B) reactive polypeptide 1 was a consequence of a delayed processing event. (iv) Analysis of herpes simplex virus 1 x herpes simplex virus 2 recombinants indicated that the determinants of type-specific immune reactivity and electrophoretic mobility of gA/B glycoproteins and g(A + B) polypeptides map near the right terminus of herpes simplex virus 1 BamHI-G. PMID- 6292508 TI - SPP1-mediated plasmid transduction. AB - The virulent Bacillus subtilis phage SPP1 transduces plasmid DNA. Plasmid transducing phages contain only plasmid DNA. Such DNA represents a concatemer of monomeric plasmid molecules with the molecular weight of mature SPP1 DNA. Biological parameters of plasmid transduction are described. PMID- 6292509 TI - Poliovirus-induced alterations in HeLa cell membrane functions. AB - Protein synthesis, amino acid uptake, membrane potential, cell volume, Na+ and K+ levels, and ATPase (Na+,K+ activated; EC 3.6.1.3) activity were investigated in control and poliovirus-infected HeLa cells. Inhibition of protein synthesis was first observed 60 min postinfection and reached a maximum at 120 min. The onset of protein synthesis inhibition coincided with a decrease in cell volume and with an elevation of ATPase activity in isolated HeLa cell membranes. Some 3 h after virus adsorption, ATPase activity was inhibited, the Na+-K+ gradient of the cell collapsed, both membrane potential-dependent tetraphenylphosphonium ion uptake and amino acid uptake were reduced, and the cell volume increased. These results provide further experimental support for the hypothesis that modification of the cell membrane plays an important role in the strategy of cytopathogenic viruses in the shutoff of host metabolism and cell death. PMID- 6292510 TI - DNA binding properties of simian virus 40 temperature-sensitive A proteins. AB - Wild-type simian virus 40 A protein (large T antigen) bound to three tandem regions of simian virus 40 DNA. The binding regions were defined by the ability of A protein to protect simian virus 40 DNA from digestion with limited (footprint assay) or excess (fragment assay) amounts of DNase I. At low concentrations, protein first bound to region I, which maps 30 to 45 base pairs to the early side of the origin of replication. At higher concentrations, A protein also protected region II and then region III. Region II spanned approximately 65 base pairs and corresponded in location to the functional origin of replication that contains a unique BglI site along with an adjacent adenine thymine-rich region. Region III was adjacent to the late boundary of region II, but its distal limit was not well defined. Twelve distinct temperature-sensitive (ts) A proteins were purified and examined for their ability to bind in regions I to III. Three classes of tsA protein were defined on the basis of thermal stability. Class I tsA protein displayed wild-type binding either with or without a heat shock. Unheated class II tsA protein exhibited wild-type binding, but after a heat shock bound very poorly to the origin of replication. Class III tsA protein was defective in its binding even without a heat shock and only protected region I. Classes II and III were coded by mutants mapping in two distinct regions of the genome. For all of the tsA proteins examined, there was a positive correlation between the thermolability of origin binding in vitro and the temperature sensitivity of these mutants for DNA replication and transcriptional autoregulation in vivo. This correlation adds support to the essential role of origin binding by A protein in viral DNA replication and early transcription repression. PMID- 6292511 TI - Antiserum specific for the carboxy terminus of the transforming protein of Rous sarcoma virus. AB - An antiserum specific for the carboxy terminus of p60src, the transforming protein of Rous sarcoma virus, was produced by immunization of rabbits with a conjugate of bovine serum albumin and the synthetic peptide NH2-Tyr-Val-Leu-Glu Val-Ala-Glu-COOH. The carboxy-terminal six amino acids of this peptide correspond in sequence to that deduced for the carboxy terminus of the p60src of the Schmidt Ruppin strain of Rous sarcoma virus of subgroup A. The p60src proteins of the several strains of Rous sarcoma virus and the cellular homolog of the viral transforming protein, p60c-src, comprise a polymorphic family of polypeptides. The anticarboxy-terminal serum reacted readily with the p60src proteins of three different strains of Rous sarcoma virus. In contrast, no precipitation of cellular p60c-src could be detected with this serum. This suggests that the viral p60src proteins have identical carboxy termini and that the carboxy terminus of cellular p60c-src may be different from that of viral p60src. The anticarboxy terminal serum reacted poorly with the subpopulation of viral p60src which is present in a complex with two cellular phosphoproteins. Apparently, the presence of the two cellular proteins interferes with the recognition of p60src by the anticarboxy-terminal serum. It seems likely, therefore, that these two cellular proteins bind to the carboxy-terminal domain of p60src. PMID- 6292512 TI - Methylation state and DNase I sensitivity of chromatin containing Moloney murine leukemia virus DNA in exogenously infected mouse cells. AB - The nature of Moloney murine leukemia virus (M-MuLV)-specific proviral DNA in exogenously infected mouse cells was studied. M-MuLV clone A9 cells, NIH-3T3 fibroblasts productively infected with M-MuLV, were used. These cells contain 10 to 15 copies of M-MuLV proviral DNA. The state of methylation of M-MuLV proviral DNA was examined by cleaving A9 cell DNA with restriction endonucleases which have the dinucleotide CpG in their cleavage sequences. Analysis with such enzymes, which recognized nine different sites in M-MuLV DNA, indicated that most if not all of the M-MuLV proviruses in A9 cells were completely unmethylated. An individual proviral integration was examined, using as probe adjacent single-copy cellular sequences. These sequences were obtained from a lambda phage recombinant clone containing an M-MuLV provirus from the A9 cells. This individual integration also showed no detectable methylation. In contrast, endogenous MuLV related sequences present in NIH-3T3 cells before infection were largely methylated. The configuration chromatin containing M-MuLV proviruses was also investigated by digesting A9 nuclei with DNase I, followed by restriction analysis of the remaining DNA. Endogenous MuLV-related DNA was in chromatin relatively resistant to DNase I digestion, whereas the majority of M-MuLV specific proviruses were in domains of intermediate DNase I sensitivity. Two proviral copies hypersensitive to DNase I digestion were identified. Analogy to the DNase I sensitivity of expressed and nonexpressed globin genes suggested that the proviral copies containing DNase I-hypersensitive sites were transcribed. PMID- 6292513 TI - Replication of mouse hepatitis virus: negative-stranded RNA and replicative form RNA are of genome length. AB - There are seven virus-specific mRNA species in mouse hepatitis virus-infected cells (Lai et al., J. Virol. 39:823-834, 1981). In this study, we examined virus specific negative-stranded RNA to determine whether there are corresponding multiple negative-stranded RNAs. Intracellular RNA from mouse hepatitis virus infected cells was separated by agarose gel electrophoresis, transferred to nitrocellulose membranes, and hybridized to positive-stranded genomic 60S [32P]RNA. Only a single RNA species of genomic size was detected under these conditions. This RNA was negative stranded. No negative-stranded subgenomic RNA was detected. We also studied double-stranded replicative-form RNA in the infected cells. Only one replicative-form of genomic size was detected. When the double-stranded RNA isolated without RNase treatment was analyzed, again only one RNA species of genomic size was detectable. Furthermore, most of the virus specific mRNAs could be released from this RNA species upon heating. These results suggest that all of the mouse hepatitis virus-specific RNAs are transcribed from a single species of negative-stranded RNA template of genomic size. PMID- 6292514 TI - Effect of aphidicolin on avian sarcoma virus replication. AB - We studied the effect of aphidicolin, an inhibitor of eucaryotic DNA polymerase alpha, on viral DNA replication and integration during the first 24 h after infection of quail embryo fibroblasts with avian sarcoma virus. In drug-treated cells, the synthesis of unintegrated linear viral DNA species was not impaired; however, the subsequent accumulation of circular viral DNA species and integrated proviral DNA was reversibly inhibited. After removal of the drug, circular viral DNA species were derived from preexisting linear viral DNA species, instead of being derived by de novo synthesis. PMID- 6292515 TI - Comparative biochemical properties of p21 ras molecules coded for by viral and cellular ras genes. AB - In earlier studies, we molecularly cloned a normal cellular gene, c-rasH-1, homologous to the v-ras oncogene of Harvey murine sarcoma virus (v-rasH). By ligating a type c retroviral promotor to c-rasH-1, we could transform NIH 3T3 cells with the c-rasH-1 gene. The transformed cells contained high levels of a p21 protein coded for by the c-rasH-1 gene. In the current studies, we have purified extensively both v-rasH p21 and c-rasH p21 and compared the in vivo and in vitro biochemical properties of both these p21 molecules. The p21 proteins coded for by v-rasH and c-rasH-1 shared certain properties: each protein was synthesized as a precursor protein which subsequently became bound to the inner surface of the plasma membrane; each protein was associated with guanine nucleotide-binding activity, a property which copurified with p21 molecules on a high-pressure liquid chromatography molecular sizing column. In some other properties, the v-rasH and c-rasH p21 proteins differed. In vivo, approximately 20 to 30% of v-rasH p21 molecules were in the form of phosphothreonine-containing pp21 molecules, whereas in vivo only a minute fraction of c-rasH-1 p21 contained phosphate, and this phosphate was found on a serine residue. v-rasH pp21 molecules with an authentic phosphothreonine peptide could be synthesized in vitro in an autophosphorylation reaction in which the gamma phosphate of GTP was transferred to v-rasH p21. No autophosphorylating activity was associated with purified c-rasH-1 p21 in vitro. The results indicate a major qualitative difference between the p21 proteins coded for by v-rasH and c-rasH-1. The p21 coded for by a mouse-derived oncogenic virus, BALB murine sarcoma virus, resembled the p21 coded for by c-rasH-1 in that it bound guanine nucleotides but did not label appreciably with 32Pi. The forms of p21 coded for by other members of the ras gene family were compared, and the results indicate that the guanine nucleotide-binding activity is common to p21 molecules coded for by all known members of the ras gene family. PMID- 6292516 TI - Molecular cloning, genomic analysis, and biological properties of rat leukemia virus and the onc sequences of Rasheed rat sarcoma virus. AB - Rasheed rat sarcoma virus (RaSV) has been shown to code for a protein of 29,000 Mr not present in replication-competent rat type C helper virus (RaLV)-infected cells. This protein is a fused gene product consisting of a portion of the RaLV p15 gag protein and the transformation-specific 21,000 Mr (p21) ras protein, which is also found in Harvey murine sarcoma virus. We now report the molecular cloning of both the SD-1 (Sprague-Dawley) strain of RaLV and the transforming ras sequences of RaSV. Heteroduplex analysis of these cloned DNAs demonstrated that the RaSV ras gene (v-Ra-ras) was inserted into the rat type C viral genome with a small deletion of RaLV genetic information in the 5' region of the gag gene and that the v-Ra-ras gene (0.72 kilobase pair) is homologous to and colinear with the p21 ras gene of Harvey murine sarcoma virus (v-Ha-ras). Restriction enzyme mapping confirmed the homology demonstrated by heteroduplex mapping, showing strong site conservation of restriction endonucleases known to cleave v-Ha-ras. Cloned v-Ra-ras DNA transformed NIH 3T3 cells, inducing the synthesis of the p29 RaSVgag-ras protein. PMID- 6292517 TI - Time-dependent maturation of the simian virus 40 large T antigen-p53 complex studied by using monoclonal antibodies. AB - Newly synthesized simian virus 40 large tumor antigen (T Ag) slowly forms a stable complex with the host tumor antigen, "p53." By the use of immunological and temporal separations and inhibition of aggregation and processing by A locus mutation, we have distinguished specific steps in the reaction sequence leading to formation of the rapidly sedimenting oligomeric complex. The monoclonal antibody PAb101 bound only a fraction of the total soluble pulse-labeled T Ag bound by antitumor serum. After a chase, all T Ag had matured to the form recognized by PAb101. All p53 in the mouse line SVA31E7 was precipitated by the T Ag-specific monoclonal antibody PAb101, even after a short pulse, and is therefore entirely bound to mature T Ag. The p53-specific monoclonal antibody PAb122 precipitates nearly all of the mature T Ag recognized by PAb101, except A locus mutant T Ag, synthesized at the nonpermissive temperature. A locus mutation inhibited entry of newly synthesized T Ag into the oligomeric greater than 28S complex of T Ag and p53. PMID- 6292518 TI - Effects of large and small T antigens on DNA synthesis and cell division in simian virus 40-transformed BALB/c 3T3 cells. AB - The roles of the large T and small t antigens of simian virus 40 in cellular DNA synthesis and cell division were analyzed in BALB/c 3T3 mouse cells transformed by wild-type, temperature-sensitive A (tsA), or tsA-deletion (tsA/dl) double mutants. Assessment of DNA replication and cell cycle distribution by radioautography of [3H]thymidine-labeled nuclei and by flow microfluorimetry indicate that tsA transformants do not synthesize DNA or divide at the restrictive temperature to the same extent as they do at the permissive temperature or as wild-type transformants do at the restrictive temperature. This confirms earlier studies suggesting that large T induces DNA synthesis and mitosis in transformed cells. Inhibition of replication in tsA transformants at the restrictive temperature, however, is not complete. Some residual cell division does occur but is in large part offset by cell detachment and death. This failure to revert completely to the parental 3T3 phenotype, as indicated by residual cell cycling at the restrictive temperature, was also observed in cells transformed by tsA/dl double mutants which, in addition to producing a ts large T, make no small t protein. Small t, therefore, does not appear to be responsible for the residual cell cycling and plays no demonstrable role in the induction of DNA synthesis or cell division in stably transformed BALB/c 3T3 cells. Comparison of cell cycling in tsA and tsA/dl transformants, normal 3T3 cells, and a transformation revertant suggests that the failure of tsA transformants to revert completely may be due to leakiness of the tsA mutation as well as to a permanent cellular alteration induced during viral transformation. Finally, analysis of cells transformed by tsA/dl double mutants indicates that small t is not required for full expression of growth properties characteristic of transformed cells. PMID- 6292519 TI - Organization of chicken DNA sequences homologous to the transforming gene of avian myeloblastosis virus. I. Restriction enzyme analysis of total DNA from normal and leukemic cells. AB - Hybridization probes consisting of cloned DNA recombinants which represent different regions of the leukemogenic sequence (amv) from avian myeloblastosis virus were used to carry out a more detailed restriction endonuclease analysis of the homologous sequences (proto-amv) present in normal and leukemic chicken DNA. The results show that four large introns interrupt the normal cellular proto-amv sequences and that there is no major rearrangement of these sequences in leukemic myeloblasts. PMID- 6292520 TI - Intracellular forms of simian virus 40 nucleoprotein complexes. IV. Micrococcal nuclease digestion. AB - The structures of DNAs present in various intracellular forms of simian virus 40 (SV40) nucleoprotein complexes were analyzed by micrococcal nuclease digestion. The results showed that the 70S SV40 chromatin was completely sensitive to nuclease digestion, whereas CsCl gradient-purified mature virion was completely resistant. Virion assembly intermediates with different degrees of virion maturation showed intermediate resistance, and three products were found: nucleosomal DNA fragments, representing the fraction of intermediates that were sensitive to nuclease; linear SV40 genome-sized DNA, representing the more mature intermediates that contained one or limited defects in the capsid shell; and supercoiled SV40, which was derived from mature virions. These digestion products, however, remained associated with capsid shells after nuclease digestion. These results were consistent with the model in which maturation of the SV40 virion is achieved through the organization of capsid proteins that accumulate around SV40 chromatin. Mild digestion of SV40 nucleoprotein complexes with micrococcal nuclease revealed the difference in nucleosome repeat length between SV40 chromatin and virion assembly intermediates. A novel DNA fragment of about 75 nucleotides was observed early in nuclease digestion. PMID- 6292522 TI - Cloning of endogenous murine leukemia virus-related sequences from chromosomal DNA of BALB/c and AKR/J mice: identification of an env progenitor of AKR-247 mink cell focus-forming proviral DNA. AB - Recombinant phages containing murine leukemia virus (MuLV)-reactive DNA sequences were isolated after screening of a BALB/c mouse embryo DNA library and from shotgun cloning of EcoRI-restricted AKR/J mouse liver DNA. Twelve different clones were isolated which contained incomplete MuLV proviral DNA sequences extending various distances from either the 5' or 3' long terminal repeat (LTR) into the viral genome. Restriction maps indicated that the endogenous MuLV DNAs were related to xenotropic MuLVs, but they shared several unique restriction sites among themselves which were not present in known MuLV proviral DNAs. Analyses of internal restriction fragments of the endogenous LTRs suggested the existence of at least two size classes, both of which were larger than the LTRs of known ecotropic, xenotropic, or mink cell focus-forming (MCF) MuLV proviruses. Five of the six cloned endogenous MuLV proviral DNAs which contained envelope (env) DNA sequences annealed to a xenotropic MuLV env-specific DNA probe; in addition, four of these five also hybridized to an ecotropic MuLV-specific env DNA probe. Cloned MCF 247 proviral DNA also contained such dual-reactive env sequences. One of the dual-reactive cloned endogenous MuLV DNAs contained an env region that was indistinguishable by AluI and HpaII digestion from the analogous segment in MCF 247 proviral DNA and may therefore represent a progenitor for the env gene of this recombinant MuLV. In addition, the endogenous MuLV DNAs were highly related by AluI cleavage to the Moloney MuLV provirus in the gag and pol regions. PMID- 6292521 TI - Altered utilization of splice sites and 5' termini in late RNAs produced by leader region mutants of simian virus 40. AB - We compared the 5' termini and splices of the late 16S and 19S RNAs synthesized by wild-type simian virus 40 and five mutants containing deletions in their late leader region. All mutants produced more unspliced 19S RNA than did wild-type virus, and in two mutants, unspliced 19S RNA constituted more than 60% of the total 19S species. The other three mutants each utilized predominantly a different one of the three spliced species of 19S mRNA. All mutants also produced decreased quantities of 16S mRNA, indicating that they may be defective for splicing both late RNAs. None of the 5' termini of the 16S and 19S RNAs made by the five mutants predominated as in those made by the wild type. Some of the mutant 5' termini were the same as those used by the wild type, whereas others were different. Although present, the major 5'-end positions used by the wild type were frequently not used as major sites by the mutants. In addition, mutants with very similar deletion endpoints synthesized RNAs with different 5' ends. Thus, downstream mutations have a pronounced effect on the location of 5' ends of the late RNAs, and there is no obvious involvement of a measuring function in the placement of 5' ends. For all mutants and wild-type virus, the 5' termini used for 16S and 19S RNAs showed major differences, with some degree of correlation found between the 5' ends and the internal splices of specific mRNA species. A model for the regulation of simian virus 40 late gene expression is presented to explain these findings. PMID- 6292523 TI - Localization of viral structural proteins in the cytoplasm and nucleus of Rous associated virus-2-infected chicken embryo fibroblasts. AB - The cellular location of viral structural proteins was carried out by immunohistochemistry and by cell fractionation. Antibody against the structural protein p27 was used in immunohistochemical reactions to demonstrate the presence of viral proteins in the cytoplasm and nucleus of Rous-associated virus 2 infected chicken cells. Localization in the nucleus was found over heterochromatic regions; in the cytoplasm it was found in discrete particulate structures. These observations were extended in cell fractionation studies in which cytoplasmic and nuclear fractions were immunoprecipitated with antibody against the viral structural proteins. PMID- 6292524 TI - Adeno-associated virus helper activity of adenovirus DNA binding protein. AB - The requirement for the adenovirus (Ad) single-stranded DNA binding protein (DBP) in the expression of adeno-associated virus (AAV) proteins was studied by specific immunofluorescent staining of infected cells and in vitro translation of RNA from infected cells. The Ad5 mutant ts125, which carries a mutation in the DBP gene, helped AAV as efficiently as the Ad5 wild type (WT) did at both the permissive (32 degrees C) and nonpermissive (40.5 degrees C) temperatures in HeLa and KB cells. Furthermore, at 40.5 degrees C ts125 was as efficient as Ad5WT was in inducing the expression of AAV proteins in a line of Detroit 6 cells which is latently infected with AAV. However, little if any AAV protein was synthesized when coinfections were carried out with Ad5WT in CV-C cells, a monkey cell line that is highly restrictive for human Ad replication unless the cells are also infected with simian virus 40. On the other hand, AAV protein was efficiently produced in CV-C cells in coinfections with the Ad5 mutant hr404, whose growth is unrestricted in CV-C cells and whose mutation also maps in the DBP gene. Finally, preparations of cytoplasmic RNA extracted from CV-C cells infected with AAV and Ad5WT or from CV-C cells infected with AAV, Ad5WT, and simian virus 40 were each capable of directing the in vitro synthesis of abundant amounts of AAV proteins in a rabbit reticulocyte lysate system. These results indicate that the abnormal DBP of ts125 still retains its helper function for AAV replication, but that the molecular feature of the DBP which relates to the monkey cell host range restriction of Ad's may also account for the observed block to AAV protein translation in CV-C cells. PMID- 6292525 TI - FBJ murine osteosarcoma virus: identification and molecular cloning of biologically active proviral DNA. AB - A 12.0-kilobase EcoRI restriction fragment containing FBJ murine osteosarcoma virus (FBJ-MSV) proviral DNA was identified in FBJ-MSV-transformed nonproducer rat cells and molecularly cloned in bacteriophage Charon 30 (lambda FBJ-1). A 5.8 kb HindIII fragment containing the entire FBJ-MSV proviral DNA was isolated from lambda FBJ-1 and subsequently subcloned in plasmid pBR322 (pFBJ-2). The DNA from recombinant plasmid pFBJ-2 was able to induce morphological transformation of rat fibroblasts in tissue culture. Transfected cells contained the p55 and p39 antigens specific for cells transformed by FBJ-MSV (T. Curran and N. M. Teich, J. Virol. 42:114-122, 1982). The organization of the FBJ-MSV provirus was analyzed by restriction endonuclease mapping, and a region of nonhomology with the helper virus was delineated. Sequences specific for this region (presumably the viral fos gene) were subcloned and used as a probe to identify related sequences present in the normal genomes of cells from a variety of mammalian species (cellular fos). A single-size (3.4 kilobases long) class of RNA hybridizing to the viral fos probe was identified in FBJ-MSV-transformed cells. PMID- 6292526 TI - Site-directed mutagenesis of the src gene of Rous sarcoma virus: construction and characterization of a deletion mutant temperature sensitive for transformation. AB - Transformation of cells by Rous sarcoma virus results from the expression of the viral src gene product, pp60src. Site-directed mutagenesis techniques have been used to construct defined deletion mutations within the src gene of Prague A strain of Rous sarcoma virus. The deletion of DNA sequences at the Bg/II restriction site in the src gene yielded both transformation-defective mutants (tdCH4, 64, and 146) and a mutant temperature sensitive for morphological transformation (tsCH119). The genome of tsCH119 contains an in-phase deletion of approximately 160 base pairs, which mapped to the immediate 3' side of the Bg/II restriction site. Upon infection of chicken cells, tsCH119 encoded a structurally altered src protein, pp53src, containing a deletion of amino acid residues 202 to 255. Immune complexes containing pp53src isolated from tsCH119-infected cells grown at 41 degrees C exhibited only 50% less tyrosine-specific kinase activity than immune complexes isolated from cells grown at 35 degrees C. pp53src immunoprecipitated from tsCH119-infected cells grown at either 35 or 41 degrees C contained phosphoserine and phosphotyrosine. We suggest that tsCH119 represents a class of mutants containing mutations mapping within a functionally important domain of the src protein, distinct from the domain specifying the protein kinase activity. PMID- 6292527 TI - Monoclonal antibodies to the transformation-specific glycoprotein encoded by the feline retroviral oncogene v-fms. AB - Monoclonal antibodies prepared to epitopes encoded by the transforming gene (v fms) of the McDonough strain of feline sarcoma virus were used to study v-fms coded antigens in feline sarcoma virus-transformed rat and mink cells. These antibodies reacted with three different polypeptides (gP180gag-fms, gp140fms, and gp120fms), all of which were shown to be glycosylated. Protein blotting with [125I]-labeled monoclonal immunoglobulin G's was used to determine the relative steady-state levels of these glycoproteins in transformed cells and showed that gp120 and gp140 were the predominant products. Immunofluorescence assays and subcellular fractionation experiments localized these molecules to the cytoplasm of transformed cells in quantitative association with sedimentable organelles. Thus, v-fms-coded glycoproteins differ both chemically and topologically from the partially characterized products of other known oncogenes and presumably transform cells by a different mechanism. PMID- 6292528 TI - Restriction enzyme analysis of partially transformation-defective mutants of acute leukemia virus MC29. AB - Restriction enzyme mapping and limited sequence analysis have been used to study the generation and genome structure of three partial-transformation mutants of avian acute leukemia virus MC29. The three mutants, td10A, td10C, and td10H, could be shown to have sustained overlapping deletions of 200, 400, and 600 base pairs, respectively, in their genomes. The precise location of the deletions was mapped within the v-myc gene of the mutants by limited sequence analysis of cloned MC29 DNA. The data obtained are discussed in terms of the effect of these deletions on the mechanism of transformation by MC29. PMID- 6292529 TI - Rauscher murine leukemia virus: molecular cloning of infectious integrated proviral DNA. AB - The integrated proviral genome of Rauscher murine leukemia virus was molecularly cloned in a bacteriophage Charon 4A vector after the proviral sequences were enriched by sequential RPC-5 column chromatography and sucrose gradient centrifugation. A recombinant DNA clone, lambda-RV-1, possessing a 12-kilobase pair EcoRI insert, was shown to contain the entire 8.8-kilobase-pair leukemia virus genome flanked by rat cellular sequences at the 5' and 3' ends. This DNA fragment was biologically active, inducing the release of virion-associated reverse transcriptase activity with as little as 10 ng of DNA insert. The virus induced XC plaque formation at high titers on NIH/3T3 and BALB/3T3 cells and demonstrated identity with the parental virus in radioimmunoassays for the highly type-specific gag gene-coded p12 protein. The molecularly cloned Rauscher murine leukemia virus should be useful in studying the molecular mechanisms involved in the transformation of specific lymphoid target cells by chronic mouse leukemia viruses. PMID- 6292530 TI - Characterization of two conformational forms of the major DNA-binding protein encoded by herpes simplex virus 1. AB - We have resolved two electrophoretic species of the major DNA-binding protein, infected cell polypeptide 8 (ICP8), encoded by herpes simplex virus 1. In pulse chase experiments, we observed the conversion of the ICP8a form, the slower migrating species, to the faster migrating form, ICP8b. Thus, the two species appear to be related as precursor-product. The conversion was not due to proteolytic cleavage, because higher concentrations of reducing agents in the sample buffer shifted the faster moving form to the slower moving species. Also, the two forms have identical peptide patterns as analyzed by partial proteolysis in sodium dodecyl sulfate. Thus, the faster moving species appears to be a conformational isomer containing intramolecular disulfide bonds. The functional significance of the two forms of the protein is discussed. PMID- 6292531 TI - Orientation and position of avian leukosis virus DNA relative to the cellular oncogene c-myc in B-lymphoma tumors of highly susceptible 15I5 X 7(2) chickens. AB - We previously reported our characterizations of the B-lymphoma tumors induced in a highly susceptible line of chickens (15I5 X 7(2)) by the avian leukosis virus RAV-1 (Proc. Natl. Acad. Sci. 78:3418-3422, 1981). We demonstrated that in greater than 90% of the tumors, the RAV-1 provirus is integrated near a cellular oncogene, c-myc. In the present study, we devised a simple approach, relying on SalI digestion, for further defining the locations and orientations of the proviruses with respect to the c-myc gene. We report here that in the great majority of cases the provirus is situated upstream from and in the same transcriptional direction as the c-myc gene--a configuration compatible with the promoter-insertion model proposed by Hayward et al. (Nature [London] 290:475-480, 1981). PMID- 6292532 TI - Survival in bilateral metachronous (asynchronous) Wilms tumors. AB - The survival of patients with Wilms tumors has improved dramatically during the last few decades. In contrast, the bilateral form of the disease, especially when the tumors are not concurrent, is still considered by many to hold a much worse prognosis. A review of 50 cases reported during the last 20 years reveals that the survival of patients with metachronous Wilms tumors has improved in parallel with survival of patients with unilateral disease and, when matched for extent of tumor spread, survivals for the last decade are almost identical to those of bilateral synchronous disease and unilateral Wilms tumors. PMID- 6292533 TI - Clinicopathological conference. Urethral valves and ureterovesical reflux associated with unsuspected findings in a nonfunctioning kidney. PMID- 6292534 TI - Mesoblastic nephroma in crossed renal ectopia. PMID- 6292535 TI - Immunologic aspects of endocrine diseases. PMID- 6292536 TI - Acute polyarthritis associated with active Epstein-Barr virus infection. AB - Nine patients with an initial onset of symptoms of acute arthritis within the preceding four weeks were enrolled in a prospective serological study with clinical follow-up for six months to two years. Four adults with chronic rheumatoid arthritis and ten healthy adults were similarly studied. Serial titers measured included antibodies to Epstein-Barr virus (EBV) antigens, group B coxsackieviruses, rubella virus, cytomegalovirus, and herpes simplex virus. Serological evidence of active EBV infection was found in four of the patients with acute arthritis, none of the patients with chronic arthritis, and one of the ten healthy adults. There was no similar correlation between acute disease and presence of antibodies to the other viruses tested. We suggest that EBV may cause acute rheumatic illnesses more commonly than is currently appreciated but is probably not involved in the etiology of typical chronic rheumatoid arthritis. PMID- 6292537 TI - Electrocardiographic abnormalities in Syrian golden hamsters with coxsackievirus B1 myocarditis. AB - Serial electrocardiograms of acute viral myocarditis were recorded from Syrian golden hamsters inoculated with Coxsackievirus B1 (CVB1). Various electrocardiographic abnormalities similar to those reported in human viral myocarditis were observed. The reciprocal ST displacement and/or T wave flattening were most frequently found, and were detected in 80% of the animals. Occasionally, other abnormalities such as atrioventricular (AV) block, left bundle branch block (LBBB) pattern or extrasystoles were recorded. A histopathological examination disclosed the most prominent lesions of myocarditis in the endocardial third of the myocardium, particularly in the interventricular septum. These lesions were more widespread in the left ventricule than in the right. A close correlation between the electrocardiographic manifestations and distribution of the lesions in the myocardium was observed. Since electrocardiography is widely used in clinical medicine, its application to experimental viral myocarditis will promote the study of viral myocarditis and the analysis of myocardial fibrosis as its sequelae. PMID- 6292538 TI - Effects of dietary sodium on brain angiotensin II receptors in spontaneously hypertensive rats. AB - The effects of dietary sodium on the characteristics of angiotensin II (A II) receptor sites in the hypothalamus-thalamus-septum-midbrain (HTSM) region were examined in spontaneously hypertensive rats (SHR) and Wistar-Kyoto rats (WKY). Twenty-four SHR and 24 WKY were divided into two groups respectively, which were maintained on high sodium diets or low sodium diets for 4 weeks, respectively. The binding capacity and affinity of the A II receptors were measured by radioreceptor assay. In WKY, the binding capacity of the A II receptors in the high sodium group was significantly lower than that in the low sodium group. On the other hand, the binding capacity of A II receptors in the high sodium group was significantly lower than that in the low sodium group. On the other hand, the binding capacity of A II receptors was not significantly different between high and low sodium groups in SHR. The secretion of arginine vasopressin (AVP) increased significantly in SHR with high sodium intake. The present results suggest that in WKY the decrease of the binding capacity of the A II receptors in the HTSM region in response to a high sodium intake serves to attenuate an osmotical stimulus to AVP secretion. However, in SHR such a regulatory mechanism as adjusting the binding capacity of the A II receptors is lacking, and this seems to be responsible, at least in part, for the enhanced secretion of AVP on the sodium loading. PMID- 6292539 TI - [Effect of reduced glutathione on endocrine and renal functions following halothane anesthesia and surgery in man]. PMID- 6292540 TI - [Fundamental and clinical studies of cefotiam in the field of oral surgery]. AB - Cefotiam (CTM) is a new synthetic cephem antibiotic developed in Japan. The results of the fundamental and clinical studies are as follows. 1. CTM showed antibacterial activity, in terms of MIC, as strong as those of cephalothin (CET) and cefazolin (CEZ) for Gram-positive cocci, and several times superior to for Gram-negative bacilli. 2. CTM serum levels approximately reached the peaks on completion of 60 minutes intravenous drip infusion of 1 g of this preparation dissolved in 5% glucose solution of 250 ml; the mean value was 65.00 micrograms/ml. Then the levels dropped rather quickly up to 180 minutes after the start of drip infusion. After that, the levels dropped gradually up to 360 minutes. 3. As for the passage of CTM in the oral tissues, satisfactory passage was observed in both maxillomandibular marrow and gingiva, which adequately exceeded MICs of the clinically isolated strains of oral infections. 4. This preparation was administered 1 g of 2 g daily by intravenous drip infusion in 18 cases of moderate or more serious infections in the field of oral surgery; the clinical efficacy rate obtained was 94.4%. 5. No manifestations of side effect were observed clinically. As for laboratory findings, 1 case of large increases in GOT and GTP (a hepatitis B antigen positive patient) and 2 cases of slight increase in GTP were observed. On the basis of these results of the fundamental and clinical studies, it was concluded that CTM is an excellent antibiotic for the treatment of oral infections. PMID- 6292541 TI - [A study of the disc sensitivity test for cefotiam]. AB - Susceptibilities to cefotiam of 103 strains of 27 bacterial species were determined by the 2-fold agar dilution method in parallel with the diameter of inhibition zone by the single-disc method, under the experimental condition established by Kanazawa. The experiments demonstrated significant correlation between MIC by the dilution method and diameter of inhibition zone in each of conventional assay of the over-night (about 16 hours) incubation, delayed assay (about 24 hours incubation), and rapid assay (about 3--4 or 5--6 hours incubation), thus confirming applicability of the single disc assay for cefotiam. Analysis of the data obtained by using cefotiam disc containing 30 micrograms revealed the primary regression equation to be: D (diameter, mm) = 25.6--10.1 log MIC (micrograms/ml) in conventional assay, D = 32.8--13.2 log MIC (micrograms/ml) in delayed assay, D = 17.2--5.8 log MIC (micrograms/ml) in 3--4 hours rapid assay and D = 21.0--7.8 log MIC (micrograms/ml) in 5--6 hours rapid assay, respectively. The range of variations in MICs estimated from the diameter of inhibition zone by the disc test was then calculated in comparison with that in MIC determined by the 2-fold agar dilution assays, as reference for the experimental errors which may be involved in the estimation of MIC of cefotiam by the single-disc assay. PMID- 6292542 TI - [Fundamental and clinical studies of ceftizoxime in obstetrical and gynecological field]. AB - This paper, is concerned with fundamental and clinical studies of ceftizoxime, a newly developed cephalosporin derivative, in the field of obstetrics and gynecology. 1. Concentrations of ceftizoxime after administration 1 g of ceftizoxime by 1 hour drip infusion were determined in genital organs in 17 patients and the exudate of pelvic dead space in 6 patients. Simulated maximal concentrations with the ratios to the simulated peak serum levels were as follows: 27.9 micrograms/g for fundal myometrium with the ratio of 48%, 36.0 micrograms/g for portio vaginalis with 62%, 17.1 micrograms/g for ovary with 29%, 15.0 micrograms/g for oviduct with 26% and 16.2 micrograms/ml for the exudate of pelvic dead space with 30%. 2. Minimal inhibitory concentrations of ceftizoxime were determined against clinically isolated organisms from female genital infectious diseases. Ceftizoxime was found to have a potent in vitro activity against Gram negative bacilli; for example, 0.1 microgram/mg or low against E. coli and K. pneumoniae. Against P. aeruginosa, P. cepacia and b. fragilis, ceftizoxime had an activity which expected to be effective in the clinical use. 3. We gave ceftizoxime to 6 patients comprising 4 patients with puerperal fever, 1 with septic abortion and 1 with tubo-ovarian abscess in daily doses of 2 to 3 g by b.i.d or t.i.d intravenous drip infusion for 4--12 days. The results of the treatment were 'excellent' in 3 patients, 'good' in 2, and 'unevaluatable' in 1. 4. Adverse reactions occurred in 2 patients who showed eruption during the medication with ceftizoxime. These patients had allergic histories due to penicillin derivatives. From the above results it is concluded that ceftizoxime is a useful drug for infections in obstetrical and gynecological field. PMID- 6292543 TI - [Susceptibility of recent clinical isolates of Pseudomonas aeruginosa and Serratia marcescens to cefotaxime, ceftizoxime, cefmenoxime, latamoxef and cefsulodin in comparison with other beta-lactam antibiotics and aminoglycosides]. PMID- 6292544 TI - [Two cases of post-therapeutic acute leukemia]. PMID- 6292545 TI - [Possibilities of gene therapy and gene transfer]. PMID- 6292546 TI - [Basic technics in DNA recombination. Restriction enzymes]. PMID- 6292548 TI - [Poliovirus vaccine and genetic engineering]. PMID- 6292547 TI - [Host-vector system in DNA recombination]. PMID- 6292549 TI - [Application of liposome fusion method to cancer therapy]. PMID- 6292550 TI - [Analysis of human carcinoantigen with monoclonal antibodies and its medical application--experimental studies]. PMID- 6292551 TI - [Neoplasma induced by retrovirus and cellular oncogene]. PMID- 6292552 TI - [Ct of hepatocellular carcinoma: its characteristics and differentiation from metastatic hepatic neoplasm]. PMID- 6292553 TI - [Investigation of 99mTc-methylene diphosphonate scintigraphy of patients with lung cancer]. PMID- 6292554 TI - [A case of hepatocellular carcinoma with intravascular growth reaching to the right ventricle and femoral vein]. PMID- 6292555 TI - [Familial amyloid polyneuropathy--evaluation of the aged patients and comparison of cases observed in Japan and in foreign countries]. PMID- 6292556 TI - [Studies of peripheral nerve conduction velocities in vibrating tool operators]. AB - Vibrating tools such as chain-saws, rock-drills, grinders and tie-tampers have been widely used in many kinds of industries in Japan, causing vibration syndrome among the operators. As is well-known, the syndrome includes disorders of the peripheral circulation in the hands, e.g. Raynaud's phenomenon, and of the peripheral and central nervous systems. As the result of preventive countermeasures to the syndrome during the last decade, the picture of the syndrome has been partly changing especially for the patients among the state forestry workers; for example, the prevalence of the peripheral nerve disorders has been relatively increasing compared with that of Raynaud's phenomenon. Peripheral nerve disorders such as numbness, paresthesia and hypesthesia in the upper limbs tend to be usually cumulative and irreversible. The disorders are very important problem of vibration syndrome from the standpoint of treatment and prevention, though there are only a few reports of the study on the disorders using electrophysiological methods. In order to make objectively clear the peripheral nerve disorders, 236 male vibrating tool operators (= Group V) were examined. The following nerve conduction velocities were measured: (1) Maximal motor nerve conduction velocities (MCV) of the median and ulnar nerves in the forearm. (2) Sensory nerve conduction velocities (SCV) of the median and ulnar nerves in the forearm and palm. (3) Residual latency (RL) of the median and ulnar nerves. Besides, working conditions and complaints relevant to vibration syndrome were examined by using questionnaires. MCVs, SCVs and RLs that were outside of mean +/- 2 S.D. of the control group (= Group C), consisting of 30 healthy men, were evaluated as abnormal. The obtained results led to the following conclusions: (1) All MCVs, SCVs and RLs of Group V were significantly less than those of the control group. In Group V, prevalence of the abnormality in each of forearm SCVs was higher than that in each of MCVs of the corresponding nerves. (2) The difference between the median and ulnar SCV in the palm and the respective SCV in the forearm of each subject was significantly greater for Group V than Group C. (Palm-forearm difference) (3) The difference between the forearm and palmar SCV of the ulnar nerve and the respective velocities of the median nerve was statistically greater for Group V than for Group C. (Ulnar-median difference) (4) Ulnar SCV in the palm of Group V was significantly related to the total operating hours of vibrating tools. (5) There was no significant relation between all the nerve conduction velocities and the white finger attack. But a significant relation was noticed between the prevalence of the abnormal values of ulnar SCV in the palm and the peripheral circulatory function score computed from the data of nail press test and cold immersion test. PMID- 6292557 TI - [Determination of free SiO2 content in deposited dusts and others by X-ray diffraction method]. PMID- 6292558 TI - [Change of serum ACE activity in patients with pneumonia]. PMID- 6292559 TI - Isolation and characterization of parainfluenza 5 virus from a dog. PMID- 6292560 TI - Serological survey of influenza A virus infection in mink. PMID- 6292561 TI - Gel diffusion test in feline infectious peritonitis with infected suckling mouse brain antigen. PMID- 6292562 TI - Children and parents and otitis media. PMID- 6292563 TI - Accessory cell function in tumor-bearing mice and effects of Corynebacterium parvum. AB - Primary in vivo production of antibody to sheep red blood cells (SRBC) was consistently suppressed in EL 4 tumor-bearing C57BL/6 mice, but the secondary response was not suppressed. This suppressed primary in vivo production of antibody was partially restored by systemic administration of Corynebacterium parvum. For investigation of the mechanism of the immunosuppression in tumor bearing mice and the effects of C. parvum, the accessory cell function of adherent cells from tumor-bearing mice and C. parvum-treated tumor-bearing mice in in vitro cultures was studied. Peritoneal and splenic cells from tumor-bearing mice were less efficient in promoting in vitro production of antibody to SRBC by macrophage-depleted normal nonadherent cells than the adherent cells from normal mice. C. parvum treatment restored the accessory cell function of splenic adherent cells from tumor-bearing mice but not that of peritoneal cells. Furthermore, adherent cells from tumor-bearing mice did not show suppressive activity against the in vitro plaque-forming cell response. PMID- 6292564 TI - Collagenolytic activity of rabbit V2 carcinoma implanted in the nude mouse. AB - The relative contribution of host cells and tumor cells to the production of collagenase and its regulation during tumorigenesis were studied with the use of a heterologous rabbit tumor-nude mouse host system. The V2 carcinoma, a malignant neoplasm of the New Zealand White rabbit, behaved as a nonmetastasizing, noninvasive tumor when implanted and grown in the inbred Swiss albino nude mouse. The extracts from both tumors contained similar levels of collagenase. Tumor explants also released enzyme into culture medium in both cases, but the rabbit tumor produced approximately 10 times more collagenase than the nude mouse. Freeze-thawing of the explants or treatment with cycloheximide markedly inhibited the appearance of enzyme in the medium from the rabbit tumor but not from the nude mouse tumor. The relative proportions of mouse- and rabbit-derived collagenase in the nude mouse tumor extracts and culture medium were determined with the use of antibodies specific for rabbit V2 tumor and mouse bone collagenases. Approximately 70% of the nude mouse tumor enzyme was derived from the rabbit tumor, and approximately 30% was derived from the mouse host. These findings indicate that the former might represent stored enzyme carried over during tumor transplantation into the nude mouse, whereas the latter might have originated from stimulation of host cells during tumorigenesis. PMID- 6292565 TI - Analysis of lymphoid cells responding to virus- and tumor-associated antigens in spleens of syngeneic mouse mammary tumor systems with low and high oncogenic potentials. PMID- 6292566 TI - [Nuclear medicine demonstration of intraocular tumors using gamma-radiating radionuclides. 1st clinical results]. AB - A non-invasive method of detecting intraocular mass lesions is described which uses 67gallium citrate and 99m-technetium pertechnetate. It was used in 33 cases. In 73.7% of the patients with malignant melanoma of the eye there was a significant increase in the storage of the tracer. There was no difference in the results obtained with 67gallium citrate and 99m-technetium pertechnetate. Small, flat and peripheral tumors may not be detected. In some cases old intraocular hemorrhages may also lead to false-positive results. In contrast to the 32P-test the method described can be used in cases with opaque media. The radiation dose is about one-tenth of that given with the 32P-test. PMID- 6292567 TI - [Ketamine anesthesia in strabismus surgery--hormone and cardiovascular tests]. AB - The effects of ketamine anesthesia on the sympathoadrenal and adrenocortical system, on plasma concentrations of somatotropine and insulin and the circulatory system were investigated in eight patients undergoing strabismus surgery. Therefore blood concentrations of adrenalin, noradrenalin, ACTH, cortisol, somatotropin and insulin as well as arterial blood pressure and heart rate were measured 30 min after premedication (I), 5 min after the initial injection of ketamine (II) and in steady state during the operation (III). Plasma adrenalin and noradrenalin levels were determined fluorimetrically by a modified trihydroxyindole technic; plasma cortisol, somatrotropin and insulin were determined by radioimmunoassay. All examined parameters--except insulin--showed significant changes after induced of anesthesia and during surgery compared to the respective pre-anesthetic values. PMID- 6292568 TI - Transport of inorganic and organic substances in the renal proximal tubule. AB - The transport through the epithelial cell layer of the renal proximal tubule proceeds in principle by passive paracellular and active transcellular transport. The active transcellular transport is mostly secondary active. This means it proceeds coupled with the flux of Na+ ions, whereby the transcellular gradient of sodium, created by the (Na+ + K+)-ATPase, located at the contraluminal cell side, provides the main driving force. Once in the cell the substances leave the other cell side by a Na+ -independent, but carrier-mediated transport system. Using microperfusion and electrophysiological techniques as well as brush border membrane vesicle preparation the Na+ -H+ countertransport and the Na+-cotransport of amino acids, phosphate, sulfate, thiosulfate, bile acids, aliphatic-aromatic monocarboxylic acids (lactate) and dicarboxylic acids was studied. Special emphasis will be given to the bidirectional transport of thiosulfate as well as to the specificity of the monocarboxylic acid and dicarboxylic acid transport system. PMID- 6292570 TI - The intrarenal renin-angiotensin-system. An immunocytochemical study on the localization of renin, angiotensinogen, converting enzyme and the angiotensins in the kidney of mouse and rat. AB - The localization of renin, converting enzyme (CE) and angiotensin II (ANG II) in the kidneys of rats and mice was investigated with immunocytochemical methods. According to the presence and specific intrarenal localization of these components of the renin-angiotensin-system (RAS) our results suggest that in addition to the well known systemic effects of the RAS, there are interactions of its components inside the kidney. These interactions may lead to the generation of an extra portion of ANG II in the renal blood stream with its target cells determined by the localization of CE at the luminal side of well defined endothelial areas. These intrarenal-intravasal reactions may or may not reinforce the action of "systemic" ANG II, generated prerenally. In addition, the existence of true intrarenal-interstitial interactions, with the different components and actions of this intrarenal RAS restricted entirely to the kidney is suggested by our results, particularly the demonstration of ANG II within epitheloid cells and the dissociation of systemic renin and ANG II from their local concentrations in renal hypertensive rats. PMID- 6292571 TI - Humoral factors and the sodium-potassium pump in low renin hypertension. AB - Recent studies suggest that sodium dependent low renin hypertension results in part from the release of a ouabain-like factor, perhaps natriuretic hormone, from the brain. This humoral factor inhibits Na+, K+-ATPase and hence the active pumping of sodium and potassium in the muscle cells of blood vessels and heart. The pump suppression causes increased contractile activity and hence increased arterial blood pressure. In the muscle cells of the blood vessels, the increased contractile activity appears to be related to membrane depolarization. PMID- 6292569 TI - Pathophysiology of human proximal tubular transport defects. AB - The generalized proximal tubular transport abnormalities comprising the Fanconi syndrome (glycosuria, generalized aminoaciduria, the proximal form of renal tubular acidosis, and increased renal clearance of phosphate, urate, calcium, magnesium, and potassium) may be ascribed to interference with "sodiumlinked" active transport. Evidence is presented that the majority of conditions known to cause Fanconi syndrome in man or experimental animals are associated with inhibitors of the renal Na-K-ATPase-ATP transport system. PMID- 6292572 TI - Molecular actions of diuretics. AB - The criteria upon which diuretics are classified is based upon their site of action within the nephron. Carboanhydrase inhibitors act in the proximal tubule, high-ceiling diuretics in the ascending loop of Henle, the thiazides in the early distal tubule and the potassium-sparing diuretics in the late distal tubule and in the collecting duct. On the molecular level diuretics do not inhibit Na+-K+ ATPase but interfere with the permeability of the tubule membranes or transport systems for certain ions and thus also influence the potential differences in the different parts of the nephron. Since carboanhydrase is located in the proximal tubule cells, not only in the cytosol but also in the brushborders and in the peritubular membranes, acetazolamide and other carboanhydrase inhibitors act on three different sites in these cells. The loop diuretics inhibit the secondary active chloride reabsorption. The receptors in this part of the nephron are stereospecific. Only the levorotatory isomere of ozolinone has active diuretic properties whereas the dextrorotatory isomere does not. Perfusion experiments of the loop of Henle with different lectins give evidence that glycoproteins containing alpha-1-fucose are involved in the reabsorption of Na+ and Cl-. Experiments on the isolated stripped rabbit colon under the condition of chloride secretion reveal striking similarities between the receptors for chloride reabsorption in the luminal cell membranes of the ascending loop of Henle and in the serosal cell membranes of the colon. The potassium-sparing diuretics amiloride and triamterene act by blocking sodium channels in the distal parts of the nephron. Thus the lumen negative potential difference decreases and (passive) potassium secretion is diminished. PMID- 6292575 TI - Epizootic coronaviral typhlocolitis in suckling mice. AB - Multiple epizootics of typhlocolitis associated with high morbidity and mortality occurred among suckling mice in an arbovirology research laboratory. Affected mice had necrosis and hyperplasia of cecal, colonic, and less often, small intestinal mucosa. Epithelial syncytia were present throughout the affected areas. Other organs generally were not involved. The etiologic agent was a coronavirus antigenically related to mouse hepatitis virus strains 1 and S. Sera from dams of affected litters and recovered animals did not contain detectable complement fixing antibody to coronavirus antigen. The lesions and the causative agent differed from previously reported coronaviral syndromes in mice. The source of the infection was not definitely found. PMID- 6292574 TI - [Case of anaerobic infection]. PMID- 6292576 TI - Eradication of Sendai pneumonitis from a conventional mouse colony. PMID- 6292577 TI - Malignant nephroblastoma in Macaca fascicularis. AB - A case of malignant nephroblastoma was identified in a 4.5-month-old female Macaca fascicularis. The age of onset, clinical history, gross and microscopic pathology, and the pulmonary metastasis were analogous to this condition as seen in man. PMID- 6292573 TI - Fibronectin--mediator between cells and connective tissue. AB - Fibronectin, previously also termed LETS-protein, is a high-molecular-weight protein (mol. w. ca. 450,000) present in the form of thin fibrils in the pericellular space of fibroblasts and other adherent cells, as well as in distinct areas of the connective tissue. A soluble form, immunologically identical and chemically at least very similar to the cell-attached protein, is found in plasma in a concentration of about 300 micrograms/ml. It is also denominated cold-insoluble globulin. The protein has affinity both to cell surfaces and to various matrix substances such as fibrin and collagen and, therefore, is capable of mediating cell attachment to these substrates. In addition, it serves as an opsonin for the phagocytosis of gelatin-containing compounds and probably is essential for the removal of soluble fibrin from the circulating blood by the reticulo-endothelial system. Bacterial cell walls are also recognized by fibronectin. A conversion of soluble fibronectin to fibrils is achieved by heparin which also enhances the binding of soluble fibronectin to cells. Heparin or, as suggested, the related heparan sulfate present on the surface of various cells, appears to function as a cofactor in the formation of pericellular fibrils. The fibronectin fibrils precipitated with heparin, compared to soluble fibronectin, show a considerably improved affinity to native collagen, especially to type III. Hyaluronic acid has an antagonistic function which, at higher concentrations, prevents the fibronectin fibrils from interacting with collagen and cell surfaces. Masking of fibronectin fibrils was also achieved by sulfated proteoglycans of cartilage. Virus-transformed fibroblasts produce less fibronectin and are less capable of maintaining surface pericellular fibrils. A reasonable explanation is that they have an elevated secretion of hyaluronic acid. The transformed cells attach only weakly to a surface and exhibit a rounded shape in contrast to healthy ones. This phenotype can be corrected to a great extent with fibronectin. It is suggested that fibronectin also influences the formation of connective tissue by accumulating collagen precursors on the surface of fibroblasts and facilitating fibrillogenesis. PMID- 6292579 TI - Management of hypoglycemia. PMID- 6292578 TI - Deposition and translocation of inhaled silica in rats. Quantification of particle distribution, macrophage participation, and function. AB - Chronic exposure to silica dust causes fibrotic lung disease. Using brief exposures, we have attempted to define the initial patterns of dust deposition, anatomical compartments through which silica is translocated, and the participation of pulmonary macrophages in clearing the inhaled dust. To accomplish this, particle distribution and translocation at the alveolar level were studied in rats exposed to aerosolized alpha-quartz. Animals were exposed to 109 mg. per cu. m. of crystalline silica in inhalation chambers for 3 hours and sacrificed at varying times after exposure. The lungs were fixed by vascular perfusion through the right ventricle and tissue blocks were prepared for transmission and scanning electron microscopy. Lungs of additional animals were lavaged to recover populations of pulmonary macrophages for in vitro studies. Scanning electron microscopy in concert with back-scattered electron imaging showed that 24 hours postexposure there was a significant decrease in the number of silica particles per unit area of alveolar duct surface when compared with lung tissue from animals sacrificed immediately after exposure. Transmission electron microscopy revealed that silica crystals had been translocated to alveolar type I cells, interstitium, and macrophages. The percentage of silica containing macrophages on alveolar surfaces increased from 36 +/- 2 per cent (mean +/- S.E.) immediately after exposure to 66 +/- 2 per cent during the 24 hours following exposure. This high percentage of macrophage participation was maintained through a 24-day postexposure period and then returned to 25 +/- 2 per cent 42 days after exposure. The percentage of silica-containing macrophages recovered by lavage were remarkably similar to those studied in situ: 24 +/- 4 per cent immediately postexposure, 62 +/- 3 per cent from 12 hours through 24 days, and 28 +/- 4 per cent 42 days postexposure. Although the percentage of macrophages with silica remained steady, the amount of silica per cell decreased during this 12-hour to 24-day period. Metabolic and viability studies of lavaged macrophages in vitro showed no differences between sham and silica-exposed animals. We propose that the events reported here represent normal, steady state clearance of a subpathogenic dose of potentially toxic particulates. PMID- 6292580 TI - Washington report. PMID- 6292581 TI - Sensitivity of proliferating human breast epithelial cells to hypotonic treatment. AB - An assay for colony-forming cells of breast epithelia derived from normal and malignant surgical specimens is described using an IMR 90 fibroblast feeder layer. Their radiosensitivity (DO: 120-172) is consistent with the proliferative origin of the colonies. Distilled water inhibits proliferation of a proportion of the colony-forming cells after a 1-minute exposure. Continued detection of colonies after 10 minutes of exposure indicates that it is an inefficient way of completely eradicating proliferating epithelial cells of normal and malignant origin. PMID- 6292582 TI - Plasma hormone responses to tamoxifen therapy and oophorectomy. AB - The effect of tamoxifen therapy on plasma hormones in the pre- and postmenopausal state was studied in a young patient with breast cancer. Tamoxifen therapy was carried out for metastatic disease prior to (premenopausal) and after oophorectomy (surgical menopause). Changes in luteinizing hormone, follicle stimulating hormone, prolactin, and estrogen were noted and were corroborated with the therapy or oophorectomy. The findings support some of the previously reported changes in those hormones that were noted in conjunction with tamoxifen therapy. PMID- 6292583 TI - Unexplained diaphragmatic paralysis: a harbinger of malignant disease? AB - The records of 103 male and 39 female patients with unexplained diaphragmatic paralysis were reviewed. A probable cause of the paralysis was not revealed by the initial history, physical examination, or review of plain chest roentgenograms. Paralysis occurred on the left in 82 patients (58%), on the right in 58 (41%), and bilaterally in two (1%). Initially, 64 patients (45%) had symptoms; dyspnea, cough, and chest wall pain were the most common. Long-term follow-up showed the best prognosis to be for patients with chest wall pain and cough (improvement in 82% and 78%, respectively); dyspnea improved in only 34% of patients with this complaint. Intrathoracic malignant lesions with phrenic nerve involvement were subsequently diagnosed in five patients (3.5%) and progressive neurogenic atrophy in one (0.7%). Roentgenographic follow-up showed return of normal diaphragmatic position in only 12 instances (9.2%). Patients with unexplained diaphragmatic paralysis are unlikely to have an underlying occult malignant or neurologic process, but recovery of diaphragmatic function is also unlikely and subsidence of related symptoms is variable. PMID- 6292584 TI - 5'nucleotidase, adenosine deaminase and purine nucleoside phosphorylase activities in acute leukaemia. AB - Three enzymes concerned in purine degradation, 5'nucleotidase (5'NT), adenosine deaminase (ADA) and purine nucleoside phosphorylase (PNP) have been measured biochemically in the bone marrow or peripheral blood blasts from 75 patients with acute leukaemia, from 18 patients with blast crisis of chronic granulocytic leukaemia and in the bone marrow and peripheral blood lymphocytes from 14 normal donors. Characteristic patterns among the different sub-types of acute leukaemia have been detected, with high ADA, low 5'NT and PNP in Thy-ALL, high 5'NT and ADA in c-ALL, high PNP and low ADA in AML. The cells in CGL blast transformation resembled the enzymatic pattern of either AML or c-ALL respectively. However, no significant correlation was found between any pair of enzymes in any group of leukaemia, normal bone marrow or peripheral blood lymphocytes studied here. PMID- 6292586 TI - [Basic immunology. 2. Immunology of tooth and oral cavity]. PMID- 6292585 TI - Difficulty in detecting in vivo extracellular infective virus in cattle naturally infected with bovine leukemia virus. AB - A search was made for the presence in vivo of infective bovine leukemia virus (BLV) in secretions and excretions as well as in several tissues from five bulls, three cows and one calf. Two of the bulls and three of the cows were natural tumour cases of enzootic bovine leukosis, another three of the bulls were natural cases of persistent lymphocytosis and the calf was a natural tumour case of juvenile bovine leukosis. BLV infection was confirmed for all cattle, except the juvenile leukosis case, by detection of BLV-specific agar gel immunodiffusion (AGID) antibodies. Cell-free preparations were made from homogenates of lymphocytes, lymph nodes, spleens, livers, intestines, urinary bladders, salivary glands, mammary glands, pools of prostate glands and testicles and feces as well as from plasma, urine and milk, by passing them through 5 micrometers membrane filters. BLV infectivity in these cell-free preparations was examined by syncytia infectivity assay using susceptible cell cultures of bovine or ovine origin. Infective BLV could not be isolated from any of these cell-free preparations of plasma, secretions, excretions and tissues, although it was isolated consistently from the in vitro cultures of viable lymphocytes obtained from BLV-infected cattle. There was no indication of BLV involvement in the case of juvenile bovine leukosis. PMID- 6292587 TI - Serum angiotensin converting enzyme in pulmonary disease. PMID- 6292589 TI - Drug-resistant herpesvirus mutants -- tough or bluff? PMID- 6292588 TI - Effects of streptozotocin-induced diabetes on calmodulin and cyclic AMP phosphodiesterase activity in rat lungs. PMID- 6292590 TI - Ca2+ transport and cell activation. AB - The role of the Ca2+ ion as a second messenger in various processes of cell activation is reviewed. In a resting cell the cytosolic Ca2+ activity is about 10(-7) M. When activated there is a Ca2+ inflow into cells or Ca2+ release from internal storage sites, which lead to an increase in the cytosolic Ca2+ activity to 10(-6)-10(-5) M. This subsequently leads to an activation of any Ca2+ sensitive processes within cells. Such activities include muscle contraction, secretion, energy metabolism, mitosis or meiosis. Pharmacological aspects of cellular Ca2+ metabolism are also discussed. PMID- 6292591 TI - [Use of laparoscopy in the diagnosis of primary malignant liver tumors]. PMID- 6292592 TI - [Diagnostic methods for herpetic encephalitis]. PMID- 6292593 TI - [Sensitivity of Enterobacteriaceae isolated from humans to 6-N,N-1',6' hexyleneformamidine-penicillanic acid (preparation HX)]. PMID- 6292594 TI - [Combined effect of 6/N,N-1',6'-hexyleneformamidine-penicillanic acid (preparation HX) and selected beta-lactam antibiotics on E. coli and Salmonella. I. Studies in vitro]. PMID- 6292595 TI - Assay of plasma angiotensin-converting enzyme activity in captopril-treated subjects. PMID- 6292596 TI - [Therapeutic effect of pivmecillinam in urinary tract infection caused by Staphylococcus saprophyticus]. PMID- 6292597 TI - [Functional morphology of the epithelium of the spiral prominence. A light, transmission and scanning electron microscope study]. PMID- 6292598 TI - Primary small cell (oat cell) carcinoma of the larynx associated with an IgD multiple myeloma. AB - Primary small cell (oat cell) carcinoma of the larynx is a rare condition. We report a case of primary oat cell carcinoma of the subglottic larynx associated with a synchronous IgD multiple myeloma (an unreported association). An increased incidence of carcinoma associated with plasma cell disorders has been reported, and the theories of this association are discussed. In a review of the reported cases, the most successful management of oat cell carcinoma of the larynx appears to incorporate a combination of radiotherapy and chemotherapy. Our case was treated with a combination of protocols used for oat cell carcinoma of the larynx and multiple myeloma. At 24 months after diagnosis, the patient is free of oat cell carcinoma, and the multiple myeloma is under control. PMID- 6292599 TI - Unilateral multiple benign mixed tumors of the parotid gland. AB - Multiple tumors of a single salivary gland in an unoperated-on patient are rare; only five have previously been reported in the world literature. The author reports the sixth case of multiple, benign mixed tumors of a unilateral parotid gland in a 61-year-old woman, discusses its management, and reviews the literature. PMID- 6292600 TI - Fracture of the clavicle following radical neck dissection and postoperative radiotherapy: a case report and review of the literature. AB - The treatment of head and neck cancer with radiotherapy and radical neck dissection has many recognized complications. Radiotherapy in therapeutic doses can produce devascularization and weakening of bone. Radical neck dissection results in altered mechanics of the shoulder girdle and a disruption of normally balanced forces acting on the clavicle. An unusual case of clavicle fracture which is considered to have resulted from an interaction of the effects of these therapies is discussed. An approach for recognizing and distinguishing this entity by its time course, and radiographic and nuclide bone scan appearance is presented. PMID- 6292601 TI - [Determination of IgM and IgG antibodies to cytomegaloviruses (CMV) using the indirect immunofluorescence technic in human serum]. PMID- 6292602 TI - Influence of cholecystokinin on hypothalamic-stalk median-eminence-extract stimulation of ACTH output from isolated pituitary cells. AB - The influence of cholecystokinin 33 (CCK33) on CRF-like stimulation of ACTH output was tested in vitro using isolated pituitary cells. ACTH was assayed using isolated adrenal cell preparations. The CRF-like material was contained in a crude acetic-acid extract of hypothalamic stalk median eminence (HSME). CCK33, in doses of 1 U, 10(-3) U, and 10(-6) U/ml cell suspension had no influence on basal or ACTH-stimulated corticosterone output from isolated adrenal cells. Isolated pituitary cells responded in a dose-related fashion of HSME extract, however, the absolute response to a given dose of HSME extract varied according to the basal (non-stimulated) output of a particular cell preparation. CCK33, in the dose range tested, had no influence on basal ACTH output. In contrast, 10(-3) U/ml oc CCK33, which corresponds to a concentration of 8 X 10(-11) M, significantly inhibited the output of ACTH from isolated pituitary cells stimulated by 0.2 equivalents of HSME. Higher concentrations of CCK33 had a variable effect. We conclude that cholecystokinin may have a role in the regulation of HSME stimulated ACTH output from the pituitary. PMID- 6292603 TI - Corticotropin-releasing factor stimulants adenylate cyclase activity in the anterior pituitary gland. AB - Ovine corticotropin-releasing factor (CRF) stimulates adenylate cyclase activity in rat anterior pituitary homogenate at an ED50 value of 70 nM. GTP increases the stimulatory effect of CRF on ]32p] cyclic AMP formation in a rat adenohypophysial particulate fraction and in bovine anterior pituitary plasma membranes. The present data show that CRF stimulates adenylate cyclase activity in the anterior pituitary gland at least partly through a guanyl nucleotide-dependent mechanism. PMID- 6292604 TI - [TRP11]-neurotensin and xenopsin discriminate between rat and guinea-pig neurotensin receptors. AB - The binding and biological activities of neurotensin and two analogues, [TRP11] neurotensin and xenopsin, in which a tryptophan replaces the neurotensin residue Tyr11, were compared in rat and guinea-pig. The binding activity of three peptides was measured as their ability to inhibit the binding of [3H]neurotensin to rat and guinea-pig brain synaptic membranes. Their biological activities were measured as their effects on the contractility of rat and guinea-pig ileal smooth muscle preparations. In binding as well as biological assays, it was found that [Trp11]-neurotensin and xenopsin were as potent as neurotensin in the rat. In contrast, the two analogues were about 10 times less potent than neurotensin in the guinea-pig. These findings reveal differences between rat and guinea-pig neurotensin receptors should be considered when comparing the activity of neurotensin analogues in assays using tissue preparations from various animal species. PMID- 6292606 TI - Proceedings of the 1982 International Narcotic Research Conference. Part I. PMID- 6292605 TI - Characterization of postsynaptic beta-adrenergic receptors and presynaptic muscarinic cholinergic receptors in the rat spleen. AB - The beta-adrenergic and muscarinic cholinergic receptors in the splenic homogenates of control and 6-hydroxydopamine (6-OHDA) treated rats were characterized. The specific binding of [3H]dihydroalprenolol (DHA) and [3H]quinuclidinyl benzilate (QNB) in the rat spleen were saturable and of high affinity and showed pharmacological specificity of splenic beta-adrenergic and muscarinic cholinergic receptors. Following 6-OHDA treatment, the Bmax value for specific [3H](-)DHA binding to the rat spleen was significantly increased by 26 percent and 22 percent compared to control at 2 and 3 weeks without a change in the Kd. In contrast, there was a 38 percent decrease in the Bmax for [3H](-)QNB in the 6-Ohda treated rat spleen at 2 and 3 weeks respectively without a change in the Kd. The Bmax value at 5 weeks was significantly greater than that at 2 or 3 weeks. The splenic norepinephrine (NE) concentration was markedly reduced by the 6-OHDA treatment at 1 to 3 weeks, while there was a significant recovery in the splenic NE concentration at 5 weeks. Thus, our results strongly suggest that we are biochemically localizing muscarinic cholinergic receptors on the sympathetic nerves of the rat spleen and that the beta-adrenergic receptors of the spleen are localized postsynaptically. PMID- 6292607 TI - Phenylmethylsulfonyl fluoride (PMSF) given systemically produces naloxone reversible analgesia and potentiates effects of beta-endorphin given centrally. AB - Intraperitoneal (IP) injection of the serine proteinase inhibitor phenylmethylsulfonyl fluoride (PMSF) produced dose-dependent analgesia in Sprague Dawley rats. AD50 was 2.9 +/- 1.4 (S.E.) mg kg-1, the analgesia was antagonized by naloxone but unaffected by atropine. PMSF significantly enhanced the analgesic effect of beta-endorphin (END) given by intracerebroventricular (ICV) infusion in rats, the enhanced END analgesia was naloxone-reversible. In Swiss-Webster mice the 24-hr LD50 value for PMSF was 215 +/- 55 mg kg-1 IP; autonomic and behavioral responses were similar to those seen in rats with ICV END. These results indicate that systemic PMSF can protect central endorphin(s) from enzymatic destruction. The significant analgesia, low toxicity, naloxone reversibility and minimal anticholinesterase effects suggest the use of PMSF as a parenteral analgesic. PMID- 6292608 TI - U-50488H, a pure kappa receptor agonist with spinal analgesic loci in the mouse. AB - U-50,488H is a chemically novel analgesic that is a potent opioid-like agent on the mouse tail flick and electrically stimulated guinea pig ileum tests. U 50,488H is a very weak competitor for naloxone binding sites in brain and ileum. However, the drug has high affinity for kappa receptor binding sites revealed by competition for EKC sites in the presence of dihydromorphine. Morphine has both supraspinal and spinal sites of action since it was a potent analgesic after both intracranial and intraspinal injections. However, U-50,488H works predominantly at the spinal level. Dynorphin may be an endogenous ligand at this site. Studies on cat dorsal horn neurons suggest that U-50,488H analgesia may be due to an increase in threshold for neuron excitation. PMID- 6292609 TI - Cross-tolerance studies distinguish morphine- and metkephamid-induced analgesia. AB - In vitro data demonstrate that metkephamid (LY127623), an analog of methionine enkephalin, has a high affinity for the delta opioid receptor, as well as the mu receptor. Data generated utilizing two in vivo measures of receptor selectivity, furthermore, indicate that metkephamid's analgesic activity is in part mediated by delta opioid receptors. The analgesic activity of metkephamid was investigated in the mouse writhing assay following chronic treatment with morphine, the prototypic mu agonist. Mice were treated chronically with increasing doses of morphine or saline and the inhibition of writhing measured in response to an acute injection of morphine or metkephamid. The dose response curve for morphine was shifted to the right 3- to 4-fold following chronic administration of morphine. In contrast, no such shift in the dose response curve for metkephamid was observed in these morphine-tolerant mice. In a further series of tests, a 50 mg/kg dose of naloxazone 20 hr prior to the assessment of morphine or metkephamid analgesia in the mouse hot plate test substantially shifted the dose-response curve for morphine to the right, while leaving the dose-response curve for metkephamid unchanged. These results suggest that delta-receptor activation contributes to the analgesia produced by metkephamid. PMID- 6292611 TI - Studies on the antinociceptive activities of mixtures of mu- and kappa-opiate receptor agonists and antagonists. PMID- 6292610 TI - Unexpected opioid activity in a known class of drug. AB - Tifluadom, although structurally a 1,4 benzodiazepine, has no affinity for the 3H flunitrazepam binding site, but is a potent displacer of 3H-bremazocine from its opioid binding site. Tifluadom is characterised as an opiate kappa-receptor agonist in vitro and in vivo with potent analgesic activity in animals and no dependence potential. PMID- 6292612 TI - In vivo evaluation of the opiate delta receptor antagonist ICI 154, 129. PMID- 6292613 TI - Comparative effects of opioid peptides on respiration and analgesia in rats. AB - Receptor mechanisms for opiate induced respiratory depression and analgesia (tail flick) were studied by the ED50 ratios and the apparent pA2 values of the interactions of naloxone with the mu-agonists morphine and D-ala2-me-phe4-met (O)ol5-enkephalin (FK-33824), and the delta-agonists D-ala2-D-leu5-enkephalin (DADL) and tyr-D-ser-gly-phe-leu-thr. The apparent pA2 values of morphine, FK 33824 and DADL for analgesia were similar, whereas the apparent pA2 values of the mu-agonists for respiratory depression were significantly lower than those of the delta-agonists. The ratio between the ED50 of FK-33824 in analgesia and respiratory depression was much lower than that of DADL. It is concluded that different receptors mediate the opiate-induced respiratory depression. One difficulty with the delta-receptors being maximally involved in this action is the high degree of antagonism shown by naloxone on the respiratory effects of the delta-agonists. PMID- 6292614 TI - Evidence for an involvement of mu-, but not delta- or kappa-opiate receptors in sympathetically and parasympathetically mediated cardiovascular responses to opiates upon anterior hypothalamic injection. PMID- 6292615 TI - Opioid agonist and antagonist bivalent ligands as receptor probes. AB - Bivalent ligands are molecules which contain two pharmacophores linked by a connecting chain (spanner). The present report describes the use of oxymorphamine (Oxy) and naltrexamine (Nal) as the opioid agonist and antagonist pharmacophores separated by a variable length spanner composed of succinyl-bis-oligoglycine. The agonist series, [CH2CO(Gly)nOxy]2, and antagonist series, [CH2CO(Gly)nNal]2, were synthesized (n = 0-4) and tested on the electrically stimulated GPI. All of the antagonist bivalent ligands (Nal) antagonized the effects of morphine, with the greatest potency enhancement (60 x) residing with the succinyl (n = 0) congener. A dramatically different SAR profile was observed in the agonist (Oxy) series where the greatest potency enhancement (17 x) occurs when n = 2. By contrast with the antagonist series the agonist bivalent ligand with n = 0 is equipotent to its monovalent agonist analogue. The significance of these results with respect to the possibility of discrete opioid agonist and antagonist recognition sites are discussed. PMID- 6292616 TI - Characterization of [3H]-etorphine binding in guinea-pig striatum after blockade of mu and delta sites. AB - The guinea-pig striatum contains an apparent homogenous population of [3H] etorphine high affinity sites (KD = 0.56 +/- 0.12 nM; Bmax = 267 +/- 47 fmoles/mg protein). The specific binding is completely abolished by 5 microM (D-Ala2, D Leu5) enkephalin whereas an important residual binding is still present after the blockade of mu and delta sites. The binding properties of these residual sites are very similar to those of the benzomorphan sites characterized in rat brain and spinal cord. From the different binding properties of kappa and benzomorphan sites, the subdivision into kappa1 (kappa sites) and kappa2 (benzomorphan sites) is discussed. PMID- 6292617 TI - Demonstration of opiate receptor sub-types in vivo. PMID- 6292618 TI - High and low affinity opioid binding sites: relationship to mu and delta sites. AB - Binding and pharmacological studies suggest a common opiate and enkephalin binding site in addition to their previously reported selective sites. This common high affinity site has tentatively been named mu1, distinguishing it from the morphine-selective site (mu2) and enkephalin-selective site (delta). The existence of this additional common high affinity site and its association with opiate and opioid peptide analgesia may help explain some pharmacological observations, such as the cross tolerance between morphine and enkephalin analgesia and the lack of cross tolerance between them in the guinea pig ileum and mouse vas deferens bioassays. PMID- 6292619 TI - Differing stereospecificities distinguish opiate receptor subtypes. AB - Paired stereoisomers of compounds active at the proposed mu, kappa and sigma classes of opiate receptors display differing stereoselectivity patterns at the receptor subtypes. The (-) isomers of cyclazocine and SKF-10047 are far more potent than the (+) isomers as displacers of [3H]dihydromorphine from receptors. However, the (-) isomers are only moderately more potent than the (+) isomers at displacing [3H]ethylketocyclazocine from kappa receptors in an assay controlled for radioligand binding to mu receptors, and the (+) and (-) isomers are similar in potency for displacement of [3H]phencyclidine (PCP) from sigma receptors. At the sigma/PCP receptor, (+) ketamine proved four times as potent as (-) ketamine, while the dioxalan derivative dexoxadrol is far more potent than its nearly inactive enantiomer levoxadrol. The results for the sigma/PCP receptor are in agreement with those of behavioral studies. Stereospecificity patterns may provide support for the concept of the opiate receptor subclasses as biochemically distinct entities. PMID- 6292620 TI - Studies on the mechanism of enkephalin receptor down regulation. AB - The association of [3H] [D-Ala2, D-Leu5] enkephalin ([3H]DADLE]) with mouse neuroblastoma cells (N4TG1) was investigated. Under identical conditions the time course, dose response curve and temperature dependence for ligand uptake were similar to those for ligand-induced receptor loss (down regulation). Uptake of [3H]DADLE was inhibited by opiate ligands as well as by the metabolic inhibitors sodium azide and 2,4 dinitrophenol. Comparison of the effects of these inhibitors on receptor binding, ligand uptake and receptor loss indicated that these cells accumulate [3H]DADLE in excess of their surface receptor number. The data suggest that receptor recycling occurs and that ligand is internalized via receptor mediated endocytosis. PMID- 6292621 TI - Solubilization in good yield of active opiate binding sites from mammalian brain. AB - Active opiate binding sites have been solubilized from mammalian brain cell membranes. The presence of 0.5-0.1 M NaCl during treatment of membranes from rat brain, human frontal cortex, and bovine corpus striatum with glycodeoxycholate or digitonin resulted in the extraction of active opiate binding sites in yields ranging up to 43%. The criteria for solubility of the sites were their inability to sediment at 10(5) x g after 2 hr and their apparent molecular weight of 3- 4 x 10(5) as determined by gel filtration. The receptors in solution resemble the membrane-bound sites with respect to saturability, stereo-specificity, sensitivity to heat and reagents, and high affinity for opioid ligands. The interaction of solubilized sites with immobilized lectins was used to demonstrate the glycoprotein nature of the opiate receptor. Soluble receptors from all species studied were retained by wheat germ agglutinin(WGA)-agarose and could be specifically eluted with N-acetylglucosamine. No retention of solubilized material was observed with eight other lectins examined, including horseshoe crab lectin, a sialic acid specific agglutinin. The receptor protein eluted from WGA columns was enriched 25-50-fold over the crude soluble fraction. PMID- 6292622 TI - Modification of the solubilized NG108-15 opiate receptor by sodium, GTP and diamide. PMID- 6292624 TI - Mu and kappa opiate binding sites in the rabbit CNS. AB - We have examined the ability of various opiates to compete with the binding of 3H etorphine (0.5 nM) in membranes from the rabbit cerebellum and thalamus. Our data suggest that greater than 80% of 3H-etorphine binding occurs at mu receptor sites in cerebellum membranes. In thalamus membranes, D-Ala2, D-Leu5-enkephalin (DADL) resolves binding of 3H-etorphine into two components. The first component accounts for about 50% of binding and may represent interaction of the radioligand with mu receptor sites. The second component is unaffected in the presence of high (1-5 microM) concentrations of DADL. The ranking of potency for opiate inhibition of the second component is ethylketocyclazocine greater than naloxone much greater than morphine much greater than DADL, suggesting it represents binding of 3H-etorphine to a kappa-opiate binding site. In the rabbit brain, the kappa-opiate binding site is particularly abundant in the thalamus followed by frontal cortex and caudate nucleus. PMID- 6292623 TI - Enkephalin and alpha-adrenergic receptors: evidence for both common and differentiable regulatory pathways and down-regulation of the enkephalin receptor. AB - Several clones of neuroblastoma-glioma NG108-15 hybrid cells were used to reveal whether the regulation of opiate receptor density interacts with the regulation of alpha-adrenergic or acetyl-choline receptors. Low density of alpha-adrenergic receptors in 3 selected clones was accompanied with similar reduction in the density of enkephalin receptors but not in muscarinic acetyl-choline receptors. Yet opiate antagonists that increased the number of opiate receptors in the parent NG108-15 cells in a stereospecific manner had no similar effect on the number of alpha-adrenergic receptors. Moreover, the stable enkephalin analogue D ala-2-methionine enkephalinamide, but not the opiate alkaloid morphine, decreased the binding of 3H-DAMEA to the membranes and induced down-regulation of enkephalin receptors. Yet DAMEA had no effect on the binding of the alpha adrenergic antagonist 3H-yohimbine. The study suggests that alpha-adrenergic and enkephalin receptors may share some common regulatory pathways but opiate peptides and antagonists selectively decrease or increase the density of enkephalin receptors, respectively, with no effect on alpha-adrenergic receptor density. PMID- 6292625 TI - Delta and sigma sites of clonal NCB20 cells do not modulate calcium uptake. AB - Opiate alkaloids and peptides are reported to inhibit 45Ca2+ binding to synaptic plasma membranes and uptake into brain synaptosomes. We have examined the effects of a number of opiates on 45Ca2+ uptake in a clonal cell line NCB20 which expresses multiple opioid binding sites. The cells express voltage-dependent calcium channels that are blocked by verapamil and nifedipine. In contrast to brain, 45Ca2+ uptake in these cells, in normal or high potassium medium, is unaffected by opiates. This difference may be due to the particular receptor types; the delta and sigma sites of these cells do not inhibit 45Ca2+ uptake. PMID- 6292626 TI - Recognition of opioid agonist and antagonist in the opioid receptor binding site. AB - By treating the rat crude synaptosomal fraction with 5,5'-dithio-bis-(2 nitrobenzoic acid), DTNB, a marked decrease of stereo-specific binding of opioid agonist (dihydromorphine or D-Ala-D-Leu-enkephalin) was observed, but there was no effect in the case of the binding of opioid antagonist (naloxone or diprenorphine). The decrease of the agonist binding in the presence of 500 microM of DTNB was nearly equal to that of 100 mM of NaCl. The ability of opioids to inhibit 3H-naloxone binding in the absence of DTNB was compared to their inhibitory potency in the presence of 500 microM of DTNB to obtain DTNB response ratio. This ratio closely correlated with sodium index of each opioid. Potency of the inactivation of the agonist binding by congeners of DTNB changed with net charge of the reagents, and 2,2'-dithiobis-(5-nitropyridine), bearing a positive charge, was most effective. These results suggest that an aliphatic sulfhydryl group, being sensitive to DTNB is located to the active center of an anionic binding site for the agonist, and controls opioid agonist binding through a proton transfer mechanism. PMID- 6292627 TI - Increase in delta, but not mu, receptors in MSG-treated rats. AB - Neonatal treatment of rats with Monosodium Glutamate (MSG) has been demonstrated to destroy cell bodies of neurons in the arcuate nucleus including the brain beta endorphin (B-END) system. The effects on opiate receptors of the loss of B-END is unknown. Seven to nine month old rats treated with MSG on the first two postnatal days and litter matched untreated control rats were decapitated and their brains dissected into several regions. Opiate receptor assays were carried out with [3H] morphine (mu receptor ligand) and [3H] DADL (delta receptor ligand) for each brain region for both MSG-treated and control rats simultaneously. Scatchard plot analyses showed a selective increase in delta receptors in the thalamus only. No corresponding change in mu receptors in the thalamus was found. The cross competition IC50 data supported this conclusion, showing a loss in the potency of morphine in displacing [3H] DADL in the thalamus of MSG treated rats. PMID- 6292628 TI - Visualization of opiate receptors and opioid peptides in sequential brain sections. AB - Autoradiographic and immunocytochemical studies were carried out on adjacent sections from formaldehyde-perfused rat brains in order to directly correlate the distribution of opiate receptors and opioid peptides. Perfusion fixation of the brains resulted in a partial loss of specific [3H]naloxone binding with essentially no change in the pharmacological properties of the remaining sites. When the distribution of sites was compared to that of enkephalin immunoreactivity in adjacent sections, striking correlations were observed in a number of areas throughout the neuraxis. Adjacent section autoradiography immunocytochemistry should provide a useful tool for relating the anatomical distribution of opiate receptor subtypes to different opioid peptide neuronal systems. PMID- 6292629 TI - The interaction of [MET5]enkephalin and [LEU5]enkephalin sequences, extended at the C-terminus, with the mu-, delta- and kappa-binding sites in the guinea-pig brain. PMID- 6292631 TI - The nature of opiate receptors in toad brain. PMID- 6292630 TI - Multiple opiate binding sites in human brain regions: evidence for kappa and sigma sites. PMID- 6292632 TI - Binding of 3H-beta-endorphin in rat brain. AB - The binding of 3H-beta-endorphin to rat brain homogenates, reported by several other laboratories, has suggested unique selective beta-endorphin binding sites. We now present additional evidence supporting the concept of distinct beta endorphin binding (epsilon) sites in rat brain. In competitive displacement studies, 3H-beta-endorphin was inhibited far better by unlabeled beta-endorphin than a variety of opiates and enkephalins. Conversely, beta-endorphin inhibited the binding of a series of 3H-labeled ligands, including dihydromorphine, ethylketocyclazocine, SKF 10,047, naloxone and D-ala2-D-leu5-enkephalin, far less potently than their corresponding unlabeled drug. Other differences were also found. Compared to 3H-dihydromorphine and 3H-D-ala2-D-leu5-enkephalin binding, 3H beta-endorphin binding was far less sensitive to the reagent N-ethylmaleimide and more sensitive to the proteolytic enzyme trypsin. The regional distribution for 3H-beta-endorphin binding was also distinct from other 3H-ligands tested. This evidence supports the concept of a distinct binding site for beta-endorphin which does not correspond to the previously defined opioid binding sites. PMID- 6292633 TI - Naloxonazine, a potent, long-lasting inhibitor of opiate binding sites. AB - Naloxazone, the hydrazone derivative of naloxone, has proven useful in studies of opiate binding site heterogeneity both in vivo and in vitro based on its long acting inhibition of high affinity, or mu1, binding sites. However, the need for high doses of naloxazone to inactivated the mu1 sites raised the possibility that its actions might result from lower concentrations of a more active compound. We now present evidence suggesting that this more active compound is the azine derivative of naloxone. In acidic solutions, approximately 35% of naloxazone, spontaneously rearranges to the azine, naloxonazine. Unlike naloxazone, naloxonazine is relatively stable in solution. It does not appreciably dissociate into naloxone and naloxazone and no additional compounds can be detected. Under assay conditions under which no azine formation can be detected, no inhibition of binding of either 3H-dihydromorphine or 3H-DADL is found after incubating tissue with naloxazone at concentrations up to 2000 nM followed by extensive washing. Naloxonazine, on the other hand, produces a potent, dose-dependent inhibition of binding which is resistant to washing. Despite the washes, naloxonazine at 50 nm abolishes high affinity binding with some inhibition seen at concentrations down to 10 nM. PMID- 6292634 TI - Differential appearance of opiate receptor subtypes in neonatal rat brain. PMID- 6292635 TI - Opioid binding sites in the midgut of the insect Leucophaea maderae (Blattaria). PMID- 6292636 TI - Supersensitivity of brain opiate receptor subtypes after chronic naltrexone treatment. PMID- 6292637 TI - In vivo binding of 3H-etorphine in morphine-dependent rats. AB - The opiate agonist 3H-etorphine was used to search for potential changes in in vivo opiate receptor binding in rats following chronic exposure to morphine sulfate. A rapid filtration method was employed to allow assessment of in vivo binding; receptor dissociation in vitro following in vivo labeling was also measured. No significant differences in total binding were seen with addicted animals, naive controls and naive animals pre-injected with morphine, at two different 3H-etorphine doses. In vitro dissociation under several conditions also yielded no differences. However, the rate of in vitro dissociation in the presence of both Na+ and a guanine nucleotide showed a small but significant decrease in dependent animals, suggesting a possible impairment of receptor effector coupling with morphine addiction. PMID- 6292638 TI - Molecular substrates of anxiety: clues from the heterogeneity of benzodiazepine receptors. PMID- 6292639 TI - Measurement of L-dihydroxyphenylalanine in plasma and other biological fluids by high pressure liquid chromatography with electrochemical detection. PMID- 6292640 TI - Allethrin interactions with the nicotinic acetylcholine receptor channel. AB - Interactions of the synthetic pyrethroid allethrin with the nicotinic acetylcholine (ACh) receptor/channel were studied in membranes from Torpedo electric organ. Allethrin did not inhibit binding of [3H]ACh to the receptor sites, but inhibited noncompetitively binding of [3H]perhydrohistrionicotoxin ([3H]H12-HTX) to the ionic channel sites in a dose-dependent manner. The inhibition constant (Ki) of [3H]H12-HTX binding in absence of receptor agonist was 30 micro M, while in presence of 100 micro M carbamylcholine it was 4 micro M. This inhibitory effect of allethrin had a negative temperature coefficient. The high affinity binding of allethrin to the channel sites of the nicotinic ACh receptor may be indicative of a postsynaptic site of action for pyrethroids, in addition to their known action on the sodium channel. PMID- 6292641 TI - Lymphocyte beta-adrenergic refractoriness induced by theophylline or metaproterenol in healthy and asthmatic subjects. AB - Beta-adrenergic refractoriness was assessed in human lymphocytes following in vivo administration of the beta-adrenergic agonist, metaproterenol, the phosphodiesterase inhibitor, theophylline, or both concomitantly, to normal and asthmatic subjects. In normal subjects both beta-adrenergic receptor number and isoproterenol stimulated cAMP response decreases during therapy with metaproterenol (59 +/- 3; 51 +/- 16% of control, respectively), theophylline (76 +/- 6; 78 +/- 16), or concomitant metaproterenol and theophylline (47 +/- 4; 69 +/- 13). The asthmatic subjects were of two types; one type responding to metaproterenol or theophylline therapy by down regulation of receptor number to zero or near zero values, and a second group of asthmatics insensitive to down regulation of receptor number. The results suggest that the induction of the refractory state is different between asthmatics and non-asthmatics, and that there may be a role for cAMP in the development of beta-adrenergic refractoriness, in vivo. PMID- 6292642 TI - Beta-endorphin modulates human immune activity via non-opiate receptor mechanisms. AB - Here we report that Beta-endorphin is a potent and efficacious suppressor of phytohemagglutinin induced T-lymphocyte blastogenesis when human leukocytes are exposed early in the course of mitogenic activation. This suppression becomes more difficult to observe, however, if blastogenesis is established by prior exposure to mitogen. Suppression by Beta-endorphin is not blocked by pretreatment with the opiate antagonist naloxone. These results, therefore, suggest that neuroendocrine modulation of human immune expression may be a peripheral physiological function of Beta-endorphin which is mediated by mechanisms distinct from traditional opiate receptors. PMID- 6292643 TI - Dimeric pentapeptide enkephalin: a novel probe of delta opiate receptors. AB - A dimeric pentapeptide enkephalin (DPE2) consisting of two molecules of [D-Ala 2, Leu 5] enkephalin linked at C-terminal leucine with ethylenediamine, (H-Tyr-D-Ala Gly-Phe-Leu-NH-Ch2)2 is a bivalent ligand for the delta enkephalin receptors of rat brain and neuroblastoma-glioma hybrid (NG108-15) cells. This new enkephalin analog shows dramatically increased affinity in radioligand assays using whole brain membranes when delta but not mu specific radioligands are employed. When membranes from NG108-15 cells are used, the dimer shows greatly increased activity irrespective of the mu or delta specificity of the tracer. The dimer DPE2 shows a four-fold, "sodium shift" in its IC50 for competition with [3H]naloxone, suggestive of agonist behavior. Agonist activity was confirmed by demonstrating that DPE2 inhibits cyclic AMP production in prostaglandin E1 stimulated NG108-15 cells, and by demonstrating very high potency in the mouse vas deferens bioassay. DPE2 binds to the same delta sites as the delta-selective monomer [D-Ala2, D-Leu5] enkephalin, since the two ligands show complete crossdisplacement. Radiolabeled 3H-DPE2 shows a five-fold higher affinity constant, a 2.5-fold higher association rate constant, and a two-fold lower dissociation rate than the monomer. These results are consistent with the hypothesis that the dimeric pentapeptide enkephalin can bridge two delta receptors. This enkephalin dimer provides a valuable new probe of opiate receptors and their organization in cell membranes. PMID- 6292644 TI - Vascular dopamine receptors: Demonstration and characterization by in vitro studies. AB - Substantial evidence has accumulated that in certain vascular beds dopamine produces its relaxant effect through stimulation of specific dopamine receptors. The goal of this review is to describe several in vitro models (perfused mesenteric vessels of the dog; renal, mesenteric, splenic, coronary and cerebral arterial strips of rabbits, dogs and cats; perfused kidney of the rat) recently developed to demonstrate such specific relaxations induced by dopamine and dopaminomimetics. On these models studies on structure-activity relationship for activation of the dopamine receptor resulted in the following order of potency for agonists: SK&F 38393 (partial agonist) greater than epinine greater than A-6, 7-DTN greater than or equal to dopamine greater than N, N-di-n-propyl-dopamine (partial agonist) greater than apomorphine (partial agonist). The dopamine receptor antagonists (+)-butaclamol, cis-alpha-flupenthixol, metoclopramide, droperidol and bulbocapnine were found to competitively antagonize dopamine induced relaxation. In addition, in two isolated organ systems (rabbit mesenteric artery, rat perfused kidney) stereospecificity of the vascular dopamine receptor was demonstrated with the isomers of butaclamol. With the development of several in vitro models demonstrating a specific antagonism against dopamine induced relaxation an important requirement for definition of a specific dopamine receptor if fulfilled according to classical pharmacological criteria. Thus, there can be do doubt on the existence of post-synaptic dopamine receptors mediating vasodilation in certain vascular tissues. PMID- 6292645 TI - ACTH and corticosterone response to naloxone and morphine in normal, hypophysectomized and dexamethasone-treated rats. AB - The effect of opiate receptors blocker naloxone on ACTH and corticosterone secretion in normal, dexamethasone-treated and hypophysectomized rats was studied. A dose-related increase in plasma corticosterone level was found at 45 min after s.c. injection of naloxone in a dose range of 0.25-2.0 mg kg-1. The rise in plasma corticosterone was preceded by a slight increase in plasma ACTH. Acute morphine administration in a relatively low dose (6 mg kg-1 s.c.) induced a significant rise in both plasma ACTH and corticosterone levels. Dexamethasone treatment was followed by low basal corticosterone level, by total inhibition of the stress response and response to morphine injection, while the response to ACTH administration was normal. Under these circumstances as well as in rats 6 days after hypophysectomy, naloxone failed to increase plasma corticosterone levels. It is concluded that a direct stimulation of corticosteroid biosynthesis in adrenal cortex is not involved in the mechanism of naloxone-induced activation of pituitary-adrenocortical function. PMID- 6292646 TI - Effect of cannabinoids on progesterone production by ovarian granulosa cells of pig and rat. PMID- 6292647 TI - CCK receptors and human neurological disease. AB - Cholecystokinin (CCK) receptor binding was measured in postmortem brain tissue of patients with Alzheimer's dementia, Huntington's chorea, and neurologically healthy matched controls. CCK binding was significantly reduced inthe basal ganglia and cerebral cortex of Huntington's patients, but was normal in the temporal and cingulate cortex of patients with Alzheimer's disease. These findings indicate that CCK receptor loss is unique to specific neurodegenerative disease(s), and that CCK may be involved in the symptoms of Huntington's disease but is not implicated in the neuropathology of Alzheimer's dementia. PMID- 6292648 TI - Insulin prevention of impaired renal calcium transport in alloxan-diabetic rats. AB - The uptake of calcium-45 was measured in slices of kidney cortex from rats with alloxan-diabetes of increasing durations. Slices from untreated diabetic rats had significantly lower slice-to-medium ratios for calcium than did slices from control rats, reflecting decreased uptake of calcium by cortical slices from diabetic rats. Institution of daily insulin therapy immediately upon confirmation of diabetes prevented this depression of calcium uptake, even though the insulin therapy was insufficient to control blood glucose levels. After four weeks of untreated alloxan-diabetes, insulin therapy did not return calcium uptake by kidney cortex slices to control levels. PMID- 6292649 TI - [Angiography and lymphography in the evaluation of radiotherapy and early detection of malignant neoplasm recurrences]. PMID- 6292650 TI - [Insulin-producing tumor of the pancreas]. PMID- 6292651 TI - [Epidemiological feature of malaria in the Ivory Coast and Upper Volta area]. AB - Several steps of the malarial epidemiological chain depend upon the environment. In West Africa, environment had no constant patterns neither in space and nor even in time. Its major discontinuities introduce disparities, at various levels, in the conditions met by the parasite for its transmission. In return, these conditions induced disparities in the immunological resistance of people. The malarial epidemiological feature is the result of the discontinuities of the environment in the Ivory Coast and Upper Volta area of the resulting epidemiological disparities. It accounts for the various possible aspects of the disease among the human populations. Malarial epidemiological feature is not only useful to understand the present situation but it is a guide of great value for any action adjusted to the needs of this area. It is also a good way to appreciate the risks resulting of the evolutions, either spontaneous or voluntary, of the natural or human environment. PMID- 6292652 TI - Infectious diseases in hospitalized renal transplant recipients: a prospective study of a complex and evolving problem. PMID- 6292653 TI - [Urinary excretion of diethyldithiocarbamate (DDC) after antabuse administration to workers chronically exposed to carbon disulfide]. AB - Djuricz's antabus test has been applied in a group chronically exposed to carbon disulphide. Sixty eight viscose industry workers hospitalized in the Clinic of Occupational Diseases, Institute of Occupational Medicine, Lodz, have been examined along with controls. In all subjects, after oral administration of antabus, the content of diethyldithiocarbamates (DDC) was determined in a 5 hr urine sample. The mean values of excreted DDC have indicated a statistically significant decrease in those chronically CS2 exposed, as compared to the controls. The dependence of DDC excretion on the duration of work in exposure has indicated a clear decrease in the values in those chronically exposed as compared to those exposed for a short time. The obtained results confirm the observations of other authors on DDC excretion decrease in CS2 exposed workers, demonstrating simultaneously some effects of chronic CS2 exposure on antabus metabolism. PMID- 6292654 TI - The X-ray investigation of the size of crystallites in quartz dusts of various fibrogenic properties. PMID- 6292655 TI - A water-borne hepatitis A outbreak in Rio de Janeiro. PMID- 6292656 TI - A study of bone proteins which can prevent hydroxyapatite formation. AB - The ability of noncollagenous bone proteins (NCBP) to inhibit calcium phosphate precipitation in vitro raises the question as to the nature and the relative efficiency of such proteins in vivo. To investigate this question NCBP from young adult sheep bones were fractionated using nondegradative techniques. Their relative activity was measured by their efficiency in preventing hydroxyapatite growth. On a weight (of protein recovered) basis, the activity is about equally divided between the Gla-containing protein, osteocalcin, and a phosphorylated protein essentially the same as osteonectin. On a molecular weight basis, the activity of the phosphorylated protein is almost three times higher than that of the Gla-containing protein. Finally, the inhibitory activity of the phosphorylated protein is alkaline phosphatase sensitive. The properties of this protein could provide a means of regulating the solubility of bone mineral and maintaining an equilibrium of calcium between bone and blood. PMID- 6292657 TI - D-Galactose dehydrogenase from Pseudomonas fluorescens. PMID- 6292658 TI - D-Erythrulose reductase from beef liver. PMID- 6292659 TI - Glycerol oxidase from Aspergillus japonicus. PMID- 6292660 TI - Glucose-6-phosphate dehydrogenase, vegetative and spore Bacillus subtilis. PMID- 6292661 TI - 6-Phospho-D-gluconate dehydrogenase from Pseudomonas fluorescens. PMID- 6292662 TI - 6-Phosphogluconate dehydrogenase from Bacillus stearothermophilus. PMID- 6292663 TI - Hydroxypyruvate reductase (D-glycerate dehydrogenase) from Pseudomonas. PMID- 6292664 TI - Spectrophotometric method for glucose-6-phosphate phosphatase. PMID- 6292665 TI - Aldehyde dehydrogenase from Proteus vulgaris. PMID- 6292666 TI - Aldehyde dehydrogenases from Pseudomonas aeruginosa. PMID- 6292667 TI - Formaldehyde dehydrogenase from Candida boidinii. PMID- 6292668 TI - Uridine diphosphate glucose-4-epimerase and galactose-1-phosphate uridylyltransferase from Saccharomyces cerevisiae. PMID- 6292669 TI - [Effect of coronavirus infection on the delayed hypersensitivity reaction]. PMID- 6292670 TI - [ATP-dependent reverse electron transport in Endomyces magnusii mitochondria]. PMID- 6292671 TI - [Effect of sodium dodecylbenzenesulfonate and antifoaming agents on the endonuclease activity of Serratia marcescens]. AB - The effect of sodium dodecylbenzene sulfonate (sulfonol) and certain froth breakers on the activity of endonuclease was studied in the cultural broth of Serratia marcescens in order to find out whether sulfonol could be used for limiting the infection. Sulfonol was found to have no effect on the cultural growth; it increased the activity of endonuclease in the cultural broth, and the peak of the activity appeared earlier than in the control medium. Propanol B-400 was shown to be the best froth breaker. PMID- 6292672 TI - Trigeminal neuralgia as a consequence of circulatory disorders in the infratemporal and pterygopalatine fossae. AB - An extensive review of the well documented experimental and clinical findings related to trigeminal neuralgia shows, that it is possible to set up an unifying theory for trigeminal neuralgia, which would explain most clinical features of the disease and also would be congruent with several other theories that have previously been presented. Thus, trigeminal neuralgia is proposed to be the result of a combination of predisposing anatomical variations, impaired arterial and venous circulation, and morphological changes in the infratemporal fossa and cavum Meckeli. The process is initiated by disturbances of dental occlusion and mandibular function, resulting in muscular hypertrophy, which finally leads to increased perineural pressure and hypoxia in the nervous tissue. Through a series of related events this results in massive afferent discharges in the trigeminal nerve, which are strong enough to break through an inhibitory gating mechanism in the nucleus caudalis, and which are experienced by the patient as a pain attack. But the tissue hypoxia, which partly has elicited the pain, will finally also be the critically delimiting factor for further signal transmission, thereby stopping the pain attack. PMID- 6292673 TI - Does an increase in the ratio of cytoplasmic NADPH to NADP+ accompanied by a decrease in the ratio of cytoplasmic NADH to NAD+ mediate the actions of insulin? PMID- 6292674 TI - Biochemical response to change in the environment and the nature of "essential" hypertension. AB - The abnormality of Na+, K+-cotransport detected in erythrocytes of patients with "essential" hypertension, appears to be an inherited biochemical disorder characteristic of the disease. This explains several features of "essential" hypertension. However, characteristics of the kinetics of this Na+ transport system are similar to characteristics of enzymes in halophilic organisms, which live in a high saline environment. Cell Na+ is increased in patients with "essential" hypertension and in halophilic organisms. This suggests that the enzyme characteristics are response mechanisms to maintain extracellular Na+ concentration within an optimal range. Based on this view two assumptions of biochemical response have been put forward which form the biochemical basis of a general biological theory. The implications of this for understanding the nature of "essential" hypertension are discussed. To explain the nature of the inheritance of the Na+, K+-cotransport defect in erythrocytes of patients with "essential" hypertension, a theory of acquired genetic transmission called "improvisation" theory is proposed. Improvisation may be the mechanism whereby new gene formation occurs, and be the mechanism for evolution of species if successful reproduction of offspring is allowed to continue. PMID- 6292675 TI - Alcohol, nutrition, and the nervous system. Alcoholic neuropathy. PMID- 6292676 TI - Differential clinical response to oestrogens after menopause. AB - An analysis of vasomotor, psychological, and physical symptoms of 136 women who were receiving piperazine oestrone sulphate (Ogen) and conjugated equine oestrogens (Premarin) after menopause has shown differences in responses which can be explained only if it is accepted that the two oestrogenic compounds have differing effects on various parts of the body. Premarin (0.625 mg) was found to be more potent at inducing withdrawal bleeding than Ogen (1.25 mg), whereas Ogen was more effective than Premarin in alleviating hot flushes and some psychological symptoms. A hypothesis involving metabolism of oestrone to the catecholamine, 2-hydroxyoestrone, is postulated, which explains why these differences occur. It is further suggested that better selection of oestrogens to suit particular postmenopausal symptoms should be encouraged when prescribing oestrogen for women after menopause. PMID- 6292677 TI - Ranitidine (ZANTAC). PMID- 6292678 TI - Peripheral facial palsy and infections- findings and problems. AB - Eighty-four patients of the Cologne University ENT Clinic with a diagnosis of idiopathic peripheral facial palsy (PEP) were examined - both clinically and virologically. In addition, examinations were carried out on 33 further PFP patients from different practising physicians (Group B) where the clinical information, however, was much less detailed. In the ENT Clinical Group (84 patients), there was a total of 12 recent virus infections (9 varicella zoster virus, 2 herpes simplex virus, 1 coxsackie B4). Proof of a recent infection depended strongly on the diagnostic prerequisites: if early and paired sera were available a virological diagnosis was possible in 32% of the cases, while in some of the other patients a recent infection could at most be suspected by the serological results. Group B with the 33 unselected patients yielded no virologically significant results. The aetiological relationship between the virological findings and PEP is discussed. PMID- 6292679 TI - Action of a polysaccharide fraction of Haemophilus influenzae lipopolysaccharide on macrophage: implication of receptor for mannosyl-polysaccharides. AB - A polysaccharide fraction (PS) was separated by mild hydrolysis from Haemophilus influenzae lipopolysaccharide. This preparation contained glycosyl-galactosyl, rhamnosyl, glucosaminyl and mannosyl residues (molar ratio: 4-1-1-2-2). It was nontoxic and immunogenic and consisted of at least one stable molecular group (fraction A; MW approximately equal to 10(6)) and an association of aggregated units (fraction B;MW approximately equal to 10(4)). This study evaluated the capacity of phagocytosis and quantitative nitroblue-tetrazolium reduction of mouse macrophages in presence of these polysaccharide fractions. After a 24-h incubation period, PS and fraction A, at 1 mg/ml, increased both phagocytosis and reduction potential of mouse peritoneal macrophages by 100%. In contrast, 1-h incubation with PS or fraction A induced a decrease of 50% in phagocytosis but no modification of NBT reduction. An identical incubation with various sugars showed that only mannosyl polymers could significantly decrease this phagocytic process. As in the case of toxic lipopolysaccharides, macrophages responded to a nontoxic preparation obtained from an endotoxin. We confirmed the role of mannosyl residues in recognition of macrophage binding receptors. Moreover, we suggest that this mannose binding ability was dependent on dose, aggregation state and molecular weight of the preparation.U PMID- 6292680 TI - Doxorubicin cardiomyopathy in children with left-sided Wilms tumor. AB - Two children with Wilms tumor of the left kidney experienced severe anthracycline cardiomyopathy after irradiation to the tumor bed and conventional dosage of doxorubicin. The cardiomyopathy is attributed 1) to the fact that radiation fields for left Wilms tumor include the lower portion of the heart and 2) to the interaction of doxorubicin and irradiation on cardiac muscle. It is recommended that doxorubicin dosage be sharply restricted in children with Wilms tumor of the left kidney who receive postoperative irradiation. PMID- 6292681 TI - Severe neurotoxicity with methyl G: CALGB experience. AB - Methylglyoxal bis-dihydrochloride is a drug that has been available for use in cancer chemotherapy since 1955. In earlier studies, it was used on a daily schedule with resulting severe gastrointestinal toxicity and myelosuppression. To avoid that toxicity, a new weekly schedule has been adopted in several Phase II studies. With the weekly schedule, new types of toxicity have been described and we report four patients who have developed a severe sensory-motor neuropathy with the chemotherapy which appears to be drug-related. PMID- 6292682 TI - [Smith-Lemli-Opitz syndrome. Case report and differential diagnosis]. PMID- 6292683 TI - [Physical analysis of the structure of dental enamel using the proton induced X ray emission method]. PMID- 6292684 TI - [Anatomo-pathological equivalents of so-called reticulosarcoma of the mouth]. PMID- 6292685 TI - Community outbreak of Norwalk gastroenteritis--Georgia. PMID- 6292686 TI - Microsomal reduction of gentian violet. Evidence for cytochrome P-450-catalyzed free radical formation. AB - The triarylmethane dye, gentian violet, is shown to undergo a one-electron reduction by the cytochrome P-450 monooxygenase system to produce a carbon centered free radical as demonstrated by direct electron spin resonance techniques. The formation of this species is inhibited by carbon monoxide and metyrapone, suggesting the involvement of cytochrome P-450. Either NADPH or NADH can serve as a source of reducing equivalents for the production of this free radical. Related triarylmethane dyes are also shown to be reduced by the monooxygenase system to form free radicals. PMID- 6292688 TI - Magnesium regulation of the beta-receptor-adenylate cyclase complex. I. Effects of manganese on receptor binding and cyclase activation. PMID- 6292687 TI - Heterogeneity of adenosine A1 receptor binding in brain tissue. PMID- 6292689 TI - Magnesium regulation of the beta-receptor-adenylate cyclase complex. II. Sc3+ as a Mg2 antagonist. AB - Sc3+ bears the same relationship to Mg2+ as La3+ to Ca2+, a similar ionic radius but increased charge. Therefore, the possibility was investigated that Sc3+ would be a Mg2+ antagonist at Mg2+ sites on the beta-adrenergic receptor-adenylate cyclase complex of the murine S49 lymphoma cell. Sc3+ is consistently much more potent than La3+ in inhibiting adenylate cyclase regardless of the mode of activation. IC50 values for Sc3+ of 10-30 microM were observed, whereas those for La3+ were about 300 microM. However, Sc3+ does not block the ability of Mg2+ to increase beta-receptor affinity for agonist nor alter agonist affinity by itself. Furthermore, Sc3+ is a weak inhibitor of the beta-receptor-mediated inhibition of Mg2+ influx. In cyc- S49 membranes, in which the catalytic subunit of cyclase cannot interact with the nucleotide-coupling protein(s), Sc3+ is as potent as in wild-type S49 membranes and again more potent than La3+. Substrate kinetics show that Sc3+, like Mg2+, modulates adenylate cyclase activity by affecting the Vmax without altering the Km for substrate. The data suggest that Sc3+ is a specific antagonist of Mg2+ at the Mg2+ site on the catalytic subunit and support the suggestion that there are two distinct sites for Mg2+ with different functions, one site on the coupling protein(s) and one on the catalytic subunit. It was also found that an apparent complex of Sc3+ and F-, ScF4-, is a potent inhibitor of adenylate cyclase, with an IC50 of 3 microM. PMID- 6292690 TI - Guanine nucleotides and monovalent cations increase agonist affinity of prostaglandin E2 receptors in hamster adipocytes. PMID- 6292691 TI - gamma-aminobutyric acid receptors modulate cation binding sites coupled to independent benzodiazepine, picrotoxin, and anion binding sites. PMID- 6292693 TI - An analysis of binding at the opioid receptor based upon an agonist/antagonist two-state model. AB - Experimental binding data for a number of opioid agonists and antagonists in homogenate of rat brain membranes at 0 degrees have been analyzed in terms of the two-state Snyder-Pert receptor model. By means of iterative nonlinear regression techniques, it was possible to evaluate the affinity constants characterizing the binding of each drug at the agonist and antagonist states of the receptor. In addition, the role of Na+ in determining the state of the receptor has been described quantitatively. The findings of this study are discussed in terms of the index, fr, which provides a theoretical measure of the fraction of receptors existing in the agonists state. The graph of fr as a function of drug concentration may be reasonably interpreted as an ideal dose-response curve which might be obtained if the drug were introduced directly at the receptor. The implications of the findings with respect to receptor operation are discussed. Although the binding constants have not been found under physiological conditions, the results indicate that the receptor mechanism remains essentially intact in the brain membrane preparation. PMID- 6292692 TI - Thermodynamic changes associated with benzodiazepine and alkyl beta-carboline-3 carboxylate binding to rat brain homogenates. PMID- 6292694 TI - The effects of exposure to unsaturated fatty acids on opiate receptors, prostaglandin E1 receptors, and adenylate cyclase activity of neuroblastoma x glioma hybrid cells. PMID- 6292696 TI - Oxytocin action. Mechanisms for insulin-like activity in isolated rat adipocytes. PMID- 6292695 TI - Modification of dose-response curves by effector blockade and uncompetitive antagonism. AB - Kinetic equations were derived for simple models of agonist-antagonist-receptor interaction and used to predict theoretical dose-response curves for the situation where there exists "receptor reserve" and a simple rectangular hyperbolic relation between tissue response and receptor occupancy by agonist, generated by an "effector chain" that links final response to receptor activation. Drug action within this chain, or an "uncompetitive" antagonism exerted by a reversible agonist that combines primarily with the active agonist receptor complex, may give the appearance of classical noncompetitive or competitive antagonism, depending upon the amount of receptor reserve and upon the rate constants involved in both agonist-receptor and antagonist-receptor interaction. The apparent potency and nature of antagonism by such an uncompetitive antagonist may depend upon the particular agonist against which it is tested. The phenomenon whereby true competitive antagonists reduce the maximal response to a partial agonist can be explained if the partial agonist, in addition to activating receptors, also acts as an uncompetitive antagonist. PMID- 6292697 TI - Kinetic determinants of benzo[a]pyrene metabolism to dihydrodiol epoxides by 3 methylcholanthrene-induced rat liver microsomes. PMID- 6292698 TI - Roles of cAMP and free fatty acids on the activity of the hexose monophosphate shunt. AB - Basal glucose utilization by isolated rat adipocytes have been found to be increased ten times in the presence of certain preparations of albumin. In these conditions the effects of several adrenergic agonists and related compounds on glucose oxidation, lipolysis and triacylglycerol synthesis in isolated fat cells have been studied. Oxidation of D(1(-14)C) glucose in rat adipocytes was almost completely inhibited by norepinephrine and isoproterenol when added to incubated fat cells. Agents able to modify intracellular AMP cyclic levels by different mechanisms display a similar ability to imitate the effect of lipolytic agents. The inhibition of glucose oxidation due to norepinephrine and isoproterenol is partially reverted by propanolol. Under the same conditions in which norepinephrine and isoproterenol markedly reduced glucose conversion to 14CO2, they stimulated lipolysis and triacylglycerol synthesis and in this case propanolol also reverted those actions. However, in these experimental conditions, norepinephrine and isoproterenol did not raise cAMP levels 10 min after hormone addition. It is concluded from these data that glucose oxidation through hexose monophosphate shunt, activation of lipolysis and triacylglycerol synthesis in isolated rat fat cells by lipolytic agents occurs by a mechanism(s) that depend(s) on intracellular free fatty acids levels. PMID- 6292699 TI - Fructose-2,6-P2, chemistry and biological function. AB - A new activator of phosphofructokinase, which is bound to the enzyme and released during its purification, has been discovered. Its structure has been determined as beta-D Fructose-2,6-P2 by chemical synthesis, analysis of various degradation products and NMR. D-Fructose-2,6-P2 is the most potent activator of phosphofructokinase and relieves inhibition of the enzyme by ATP and citrate. It lowers the Km for fructose-6-P from 6 mM to 0.1 mM. Fructose-6-P,2-kinase catalyzes the synthesis of fructose-2,6-P2 from fructose-6-P and ATP, and the enzyme has been partially purified. The degradation of fructose-2,6-P2 is catalyzed by fructose-2,6-bisphosphatase. Thus a metabolic cycle could occur between fructose-6-P and fructose-2,6-P2, which are catalyzed by these two opposing enzymes. The activities of these enzymes can be controlled by phosphorylation. Fructose-6-P,2-kinase is inactivated by phosphorylation catalyzed by either cAMP dependent protein kinase or phosphorylase kinase. The inactive, phospho-fructose-6,P,2-kinase is activated by dephosphorylation catalyzed by phosphorylase phosphatase. On the other hand, fructose-2,6 bisphosphatase is activated by phosphorylation catalyzed by cAMP dependent protein kinase. Investigation into the hormonal regulation of phosphofructokinase reveals that glucagon stimulates phosphorylation of phosphofructokinase which results in decreased affinity for fructose-2,6-P2 appears to be due to the decreased synthesis by inactivation of fructose-2,6-P2,2-kinase and increased degradation as a result of activation of fructose-2,6-bisphosphatase. Such a reciprocal change in these two enzymes has been demonstrated in the hepatocytes treated by glucagon and epinephrine. The implications of these observations in respect to possible coordinated controls of glycolysis and glycogen metabolism are discussed. PMID- 6292700 TI - [Interactions of subunits of DNA gyrase]. AB - Interaction of DNA gyrase A- and B-subunits during the process of DNA supercoiling was studied. For this purpose a E. coli Cour-1 mutant resistant to coumermycin and containing a mutation in the B-subunit of DNA gyrase was isolated and the influence of the DNA gyrase A-subunit specific inhibitor-nalidixic acid on DNA supercoiling by wild-type and mutant enzymes was investigated. It turned out that the enzyme from the Cour-1 mutant strain was more sensitive to nalidixic acid than the DNA gyrase from the wild-type strain. Hence, the mutation affecting the B-subunit is capable to change A-subunit properties. That makes it possible to draw the conclusion about a close structural interaction of DNA gyrase subunits during DNA supercoiling. PMID- 6292702 TI - [Construction of hybrid plasmids containing the yeast replicator]. AB - In Saccharomyces cerevisiae strain 6-1G-P188 about 10 per cent of rRNA genes exist as extrachromosomal copies of rDNA repeating units. These extrachromosomal copies can be isolated as covalently closed molecules with lengths around 3mu. We have constructed a set of hybrid plasmids containing the bacterial vector pBR325, the LEU2 gene of yeast encoding beta-isopropylmalatedehydrogenase and various EcoRI restriction fragments of the 3mu DNA. We have tested the ability of our hybrid plasmids to transform LEU2 strain DC5 to leucine prototrophy. One of the plasmids Rcp21/11 transforms DC5 at the frequency comparable with that obtained with YEp13, containing the 2mu DNA replication origin. The 2400 bp EcoRI-B fragment of the 3mu DNA in Rcp21/11 carries a gene for 5S rRNA and two spacers. Our results on transformation experiments allow un to suggest that this EcoRI fragment also carries the 3mu DNA replication origin. Yeast transformants containing this plasmid are highly unstable but during the prolonged growth in selective conditions the stabilization of the LEU+ phenotype is observed being most likely a result of integration of Rcp21/11 into the yeast chromosome. PMID- 6292701 TI - [Study of spatial organization of the RNA of phage MSZ using nucleases specific for the secondary structure]. AB - The limited hydrolisis of bacteriophage MS2 RNA by nuclease S1 and ds-specific snake venom RNase was studied in a wide range of ionic strength, at different pH, after heating and (slow and fast) cooling and at various enzyme-substrate relations. It was shown that the RNA has exposed hydrolisis sites for both nucleases. The localizations of these sites are very specific and are not altered in all conditions studied. The hydrolisis rate was changed in some conditions, at that the fragments patterns in denaturing electrophoresis did not move. It was supposed that the RNA has strongly predetermined and predominant conformation which could not be altered by strong influences. PMID- 6292703 TI - [Dibromoethylacetate - a new agent for fixing the unwound regions of DNA. Denaturation maps of mitochondrial DNA from the rat liver]. AB - On the basis of the reaction of dibromoethylacetate with adenine and cytosine at the uncoiled regions of DNA it was possible to fix these molten regions (the degree of denaturation was about 16%) for rat liver mitochondrial DNA. Modification of all accessible adenine and cytosine residues in 0.14 M Na acetate, pH 5.65, was completed at 70-80 degrees C in 15-20 min. Using the computer orientation for the set of fixed BamHI-fragments and the linear molecules of full length the denaturation map of mtDNA was constructed, the GC content of the molten regions was about 28%. The dibromoethylacetate is a perspective agent for screening DNA with a low content of destabilizing regions (for instance, DNA of malignant cells). PMID- 6292704 TI - [Stimulation of linear DNA transcription by DNA gyrase]. PMID- 6292705 TI - The binding and activation of the Clr-Cls subunit of the first component of human complement. AB - The value of the functional affinity constant between 125I-labelled Clq and the Clr-Cls tetramer (when free in solution) in the formation of Cl was found to be 3.6 X 10(7) M-1. When Clq was bound to activating immune complexes, the value of K was about 10-fold higher before initiation of activation and there was a further two to three-fold rise as activation proceeded. The addition of an excess of unlabelled Clq increased the rate of activation of 125I-labelled Cl, suggesting an interaction between Clr-Cls and two neighbouring Clq molecules. It is suggested that the tetramer Clr-Cls may bind bivalently to Clq when free in solution, but on binding to activating complexes, one of the Clr-Cls binding sites is detached from Clq and becomes bound to a site on the complex. The resultant spatial rearrangement of the Clr molecules within the tetramer may be optimal for autocatalytic activation of Clr. PMID- 6292706 TI - [Molecular action mechanism of benzodiazepines. Intensification of endogenous control mechanisms for sleep, fear, muscle tone and spasmodic activity]. PMID- 6292708 TI - Further evidence that ultraviolet radiation-enhanced reactivation of simian virus 40 in monkey kidney cells is not accompanied by mutagenesis. AB - Can simian virus 40 (SV40) be used to detect mutagenic DNA repair in cultured mammalian cells? The published evidence from different laboratories are in direct conflict. In order to decide between the conflicting evidence, we conducted experiments in two separate laboratories using experimental protocols similar to those previously used to investigate mutagenic repair with viral probes. Mutagenesis in SV40 virus stocks obtained by infecting ultraviolet (UV) irradiated or unirradiated CV-1 monkey kidney cells with UV-irradiated or unirradiated temperature-sensitive SV40 mutant tsB201 was investigated. The frequency of reversion of the ts mutant to phenotypically wild-type virus was determined by assaying the virus stocks at permissive (33 degrees) and non permissive (39 degrees) temperatures. These data show that (a) the reversion frequency for unirradiated virus propagated in irradiated cells was more than that in unirradiated cells; (b) irradiated virus gave more reversion than unirradiated virus in unirradiated and irradiated cells; and (c) irradiated virus had a lower reversion frequency in irradiated cells than in unirradiated cells. Reactivation experiments carried out in parallel; with the mutagenesis showed enhanced reactivation in UV-irradiated SV40 in UV-irradiated CV-1 cells. We conclude that enhanced reactivation of UV-irradiated SV40 was not mutagenic in monkey kidney cells. PMID- 6292707 TI - [Endocrine pancreas tumors. Progress in diagnosis and surgical therapy]. PMID- 6292709 TI - Effect of benzo[a]pyrene on sister-chromatid exchange in fetal hamster liver exposed in utero. AB - Benzo[a]pyrene between 50 and 125 mg/kg administered maternally caused a dose related increase in sister-chromatid exchange in fetal hamster liver cells. There was no difference on days 11, 13 and 15 of gestation in the sensitivity of fetal liver to benzo[a]pyrene. PMID- 6292710 TI - Measurement of spontaneous mutation rates at the Na+/K+ ATPase locus (ouabain resistance) of human fibroblasts using improved growth conditions. AB - The mutation rate for the Na+/K+ ATPase locus (ouabain resistance, OuaR) in mammalian cells in culture has been reported to be 10-100-fold lower than the mutation rate of other gene loci in culture, such as the hypoxanthine phosphoribosyl transferase (HPRT) locus. Determination of the mutation rate to ouabain resistance is sensitive to culture conditions and the concentration of ouabain used to select mutants. Our improved growth conditions for human cells have permitted absolute cloning efficiencies of 70-90% and population doubling times of 16-17 h with both normal human diploid fibroblasts, KD, and their chemically induced neoplastic derivative, Hut-11A. Ouabain at 10(-7) M was found to be adequate to select for resistant (OuaR) mutants with an absolute recovery efficiency of 54-102%. Under these conditions, the mutation rates to ouabain resistance for human cells were measured and found to be 1-8.5 X 10( 7)/cell/generation for KD cells and 6-13 X 10(-7)/cell/generation for Hut-11A cells. These rates are 5-25 times higher than previously reported for human cells. Improved growth and the use of a lower concentration of ouabain for selection may allow for the increased recovery of OuaR mutants and an improved estimate of the mutation rate at this locus, which is only 2-10-fold less than the mutation rate at the HPRT locus in the same cells. PMID- 6292711 TI - Morphometric analyses of human muscle fiber types. AB - Fibers from the m. vastus lateralis of 10 middle-aged men were classified at ultrastructural level according to the appearance of the sarcomeric M-band. The Z band widths had a two-peak distribution. One peak was due to type 1 fibers (mean 125 +/- 11 nm), the other to type 2 fibers. This latter could be separated into type 2A (101 +/- 9 nm) and type 2B (86 +/- 8 nm). About 83% of the fibers would have been correctly classified on the basis of the Z-band width alone. Mitochondrial volumes differed (type 1 5.6 +/- 0.8, 2A 4.0 +/- 0.8, and 2B 2.8 +/ 0.8%). However, only one third (37%) of the fibers would have been correctly classified if sorted according to this parameter. Mitochondrial volumes in the different fibers were correlated to mitochondrial enzymes, while fiber sizes and numbers were correlated to cytoplasmic variables. The correlations appeared mainly after a training program, suggesting that the relationships between structural and functional parameters are more obvious after adaptation to higher functional demands. PMID- 6292712 TI - Effect of ketoconazole on the initial stages of germ tube formation by strains of Candida albicans. PMID- 6292713 TI - Purification, characterization, substrate and inhibitor specificity of adenosine kinase from several Eimeria species. AB - Ribonucleosides of some pyrazolo [3,4-d] pyrimidines have been shown to be potent anticoccidial agents. To investigate their interactions with adenosine kinase, this enzyme was purified by affinity chromatography from the sporulated oocysts of 3 avian coccidia, Eimeria tenella, E. acervulina and E. brunetti as well as from chicken liver. Comparative studies revealed several differences among the enzymes. Magnesium appeared not to be inhibitor of the E. tenella enzyme but did inhibit the enzymes from the other three sources. ATP in excess of the magnesium concentration strongly inhibited the E. brunetti enzyme but had only a small effect on the other enzymes. The chicken liver enzyme utilized a broader variety of triphosphate donors than did any of the enzymes from Eimeria species. ATP, dATP, GTP, dGTP and ITP was the best substrates. Studies with pyrazolo [3,4-d] pyrimidine nucleosides revealed two groups of enzymes with similar inhibitor specificities, the chicken liver and E. Acervulina vs. the E. tenella and E. brunetti enzyme. This grouping roughly correlates with the in vivo anticoccidial specificity of these compounds. Substrate specificity studies using two 4 substituted pyrazolo [3,4-d] pyrimidine ribonucleosides (ethylthio- and cinnamylthio-), which have shown potent anticoccidial activity in vivo, revealed that each served as a substrate for the enzymes from E. tenella and E. acervulina. The E. tenella enzyme was the more efficient at the phosphorylation of those compounds. However, only the ethylthio- compound was detectably phosphorylated by the enzyme from E. brunetti. In contrast to the inhibitor specificity, the substrate activities of these nucleosides do not correlate well with their in vivo anticoccidial activity. PMID- 6292714 TI - Identification of maxicircle DNA sequences in Leishmania Tarentolae that are homologous to sequences of specific yeast mitochondrial structural genes. AB - Sequences homologous to the yeast mitochondrial structural genes for cytochrome oxidase subunits I and II, ATPase 6 and cytochrome b were identified on the kinetoplast DNA maxicircle molecule by low stringency hybridization of maxicircle blots with heterologous probes derived from mitochondrial DNA of yeast petite mutants. No hybridization was observed with the yeast ATPase 9 gene probe. The relative extent of base sequence mismatch was determined by melting of the heterologous hybrids. Candidates for the transcripts of these presumptive structural genes were proposed with reference to the transcriptional map of the maxicircle of Leishmania tarentolae. These results provide the first indication that maxicircle DNA specifies information for a limited number of conserved mitochondrial gene products similar to those already described for other eukaryotic cells. PMID- 6292716 TI - Ocular findings of acute cytomegalovirus infection. PMID- 6292715 TI - Immunology of autoimmune thyroid diseases. PMID- 6292717 TI - Retroviruses produced by hybridomas. PMID- 6292718 TI - Mechanism of action of calcium-channel-blocking agents. PMID- 6292719 TI - Stimulated human phagocytes produce cytogenetic changes in cultured mammalian cells. PMID- 6292720 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 1-1983. A 37-year-old schizophrenic woman with hypoglycemic episode. PMID- 6292722 TI - Glucocorticoid regulation of protein processing and compartmentalization. PMID- 6292721 TI - Polymorphism in the 5' flanking region of the human insulin gene: a genetic marker for non-insulin-dependent diabetes. AB - We sought to determine whether differences in the human insulin gene or its immediate flanking sequences could be found in diabetes. Peripheral leukocyte DNA from 217 unrelated persons, including blacks, whites, and Pima Indians, was analyzed by restriction-enzyme digestion, blotting to nitrocellulose filters, and hybridization to cloned [32P]insulin-gene probes. A region of length variation including deletions (0.1 to 0.2 kilobase pairs) or insertions (0.6 to 5.5 kb) of DNA was found only in the immediate 5' flanking region in 33 per cent of the genes examined. A 1.6-kb insertion accounted for 80 per cent of the polymorphism. This variant was found more often in subjects with non-insulin-dependent diabetes than in nondiabetics, regardless of race (P = 0.011). Length polymorphism in the 5' flanking region of the insulin gene may provide a genetic marker for non insulin-dependent diabetes. PMID- 6292723 TI - Luteinizing hormone release from dissociated pituitary cells by dimerization of occupied LHRH receptors. PMID- 6292724 TI - Characterization of a growth hormone-releasing factor from a human pancreatic islet tumour. PMID- 6292725 TI - A transposon in Streptococcus faecalis with fertility properties. PMID- 6292726 TI - A voltage-gated ion channel model inferred from the crystal structure of alamethicin at 1.5-A resolution. AB - The crystal structure of alamethicin in nonaqueous solvent has been determined, and refined at 1.5-A resolution. The molecular conformation of the three crystallographically independent molecules is largely alpha-helical with a bend in the helix axis at an internal proline residue. The helix structure is highly amphipathic as most of the solvent-accessible polar atoms lie on a narrow strip of surface parallel to the helix axis. Molecular models for the voltage-gated ion channel, with n-fold symmetry and based on the molecular conformations observed in the crystal, are characterized by strong surface complementarity, a hydrophilic interior and a hydrophobic exterior. The channel structures are stabilized by a hydrated annulus of hydrogen-bonded glutamine residues which produce the greatest restriction in the channel diameter. PMID- 6292727 TI - DNA strand breaks and ADP-ribosyl transferase activation during cell differentiation. PMID- 6292728 TI - Role of DNA breaks and ADP-ribosyl transferase activity in eukaryotic differentiation demonstrated in human lymphocytes. PMID- 6292729 TI - Allelic polymorphism and complexity of the genes for HLA-DR beta-chains--direct analysis by DNA-DNA hybridization. PMID- 6292730 TI - Nucleotide sequence of gamma delta resolvase gene and demonstration that its gene product acts as a repressor of transcription. PMID- 6292731 TI - Synchronized bursting of CA1 hippocampal pyramidal cells in the absence of synaptic transmission. PMID- 6292732 TI - A protein with kinase and phosphatase activities involved in regulation of tricarboxylic acid cycle. PMID- 6292733 TI - Evidence for the direct involvement of DNA replication origin in synthesis of late SV40 RNA. PMID- 6292734 TI - CA2+-free perfusion of rat heart reveals a (Na+ + K+)ATPase form highly sensitive to ouabain. PMID- 6292735 TI - Minute virus of mice inhibits cell transformation by simian virus 40. PMID- 6292736 TI - Hypertension and inhibition of the sodium pump: a strong link but in which chain? PMID- 6292737 TI - Activation of a cellular oncogene by DNA rearrangement: possible involvement of an IS-like element. AB - The cellular oncogene c-mos is rearranged in a mouse myeloma and the tumour mRNA contains transcripts hybridizing with a v-mos probe. The rearranged gene (rc-mos) was cloned in lambda phage and shown to transform mouse fibroblasts in transfection assays, rc-mos differs from its progenitor, c-mos, only at the 5' end of the gene, where c-mos sequences have been substituted by a novel cellular DNA fragment. This fragment contains a 159-base pair (bp) insertion sequence (IS) like element localized immediately 5' to the junction with c-mos. This is the first demonstration in a non-virally-induced tumour of activation of a cellular oncogene by a mechanism possibly involving DNA transposition. PMID- 6292738 TI - A circulating inhibitor of (Na+ + K+)ATPase associated with essential hypertension. PMID- 6292739 TI - Activation of a cellular transforming gene in tumours induced by Abelson murine leukaemia virus. PMID- 6292740 TI - Involvement of alpha2-adrenoceptors of nucleus tractus solitarius in baroreflex mediated bradycardia. AB - Microinjections of noradrenaline and clonidine into nucleus tractus solitarius produced dose dependent bradycardia without significant decrease in blood pressure in chloralose anaesthetized cats. Phenylephrine failed to produce any significant alteration of heart rate or blood pressure. Piperoxan microinjection into nucleus tractus solitarius elicited a mild but significant tachycardia and could also block the noradrenaline and clonidine responses. Phenoxybenzamine however neither affected resting heart rate and blood pressure nor antagonized the responses of noradrenaline and clonidine. Guanethidine pretreatment of nucleus tractus solitarius also abolished the clonidine response. Baroreceptor reflex activation induced bradycardia was inhibited by yohimbine or piperoxane injected into the cisterna magna or microinjected bilaterally into the nucleus tractus solitarius. Pretreatment of nucleus tractus solitarius with phenoxybenzamine by either route, did not affect the reflex bradycardia. It is concluded that the alpha-adrenoceptors of nucleus tractus solitarius involved in the decrease in heart rate during baroreceptor activation are alpha2 in nature. PMID- 6292741 TI - Augmentation of vagal reflex bradycardia be central alpha2-adrenoceptors in the cat. AB - Vagal reflex bradycardia was induced in anaesthetized cats with high level spinal axotomy by electrical stimulation of either the carotid sinus nerves or a depressor nerve. In both preparations reflex bradycardia increased with the rate of stimulation. Injection of 1 microgram/kg clonidine into a lateral cerebral ventricle augmented reflex bradycardia in response to carotid sinus nerve stimulation while the same dose of clonidine was ineffective when given intravenously. The antagonistic effect of intracerebroventricular yohimbine (50 micrograms/kg) indicated that the effect of clonidine was due to its alpha2 agonistic action. In contrast to carotid nerve stimulation the reflex bradycardia in response to depressor nerve stimulation was affected neither by intracerebroventricular injection of clonidine (2 micrograms/kg) nor by yohimbine (100 micrograms/kg). It is concluded that in the cat, the function of the central parts of the baroreceptor reflex which originate from the carotid sinus area is augmented by stimulation of alpha2-adrenoceptors while the function of those parts originating from the aortic area is not. PMID- 6292742 TI - [125I]-HEAT:fifty percent of the ligand can bind to the alpha1-adrenoceptors with extremely high affinity. AB - [125]-HEAT, 125iodo-2-[Beta-(4-hydroxyphenyl)-ethyl-aminomethyl]tetralone, is a novel alpha1-adrenoceptor ligand which labels alpha1-adrenoceptors in peripheral tissues as well as in the central nervous system. Using the technique of ligand saturation by receptors, we find that only 50% of the 125I-labeled HEAT molecules bind with high affinity to receptors from a variety of tissues. This was observed with partially purified rat brain membranes and highly purified rat liver plasma membranes in the absence or presence of sodium ion (as NaCl, 150 mM) which stimulated 125I-HEAT binding, by increasing the affinity. If the bindability of [125I]-HEAT is taken into account, KD values as low as 7-8 pM (at 30 degrees C) are found in equilibrium binding experiments and optimally stimulating concentrations of sodium ion. The limited high affinity binding of [125I]-HEAT could not be explained by radiochemical impurities. Instead, we suggest that only one enantiomer of the racemic ligand is preferentially bound to the receptors with a KD in the picomolar range. Since the enantiomers are in dynamic equilibrium in solution (via keto-enol tautomerism) [125-I]-HEAT is a unique radioligand which makes it unlikely that the respective isomers can be separated by successive depletion with receptors. PMID- 6292743 TI - Does presynaptic regulation of sympathetic transmission occur within a limited range of neuronal activity? AB - Recent reports indicate that the presynaptic negative feedback mechanism involved in regulating sympathetic neurotransmission is only functional within a narrow range of neuronal activity. The specific aim of this report is to examine the release of sympathetic transmitter at varying frequencies of stimulation, and the effects of alpha-adrenoceptor antagonists on the overflow. The experiments were carried out in the 3H-noradrenalin-labeled heart of the guinea pig. The overflow of tritium (per pulse) increased from a stimulation frequency of 0.125 to 10 Hz and declined at 30 Hz. Phentolamine or yohimbine produced facilitation of the overflow from 0.125 to 10 Hz; the effect was more pronounced at lower frequencies of stimulation. We conclude that the presynaptic alpha-adrenoceptor-mediated negative feedback mechanism operates over a wide range of cardiac sympathetic nerve activity. PMID- 6292744 TI - Evidence of multiple receptor sites within the putative calcium channel. AB - [3H]-Nimodipine a potent calcium channel blocker, binds to an apparently homogenous population of receptors in guinea-pig brain membranes (KD=0.62 nM, Hill coefficient or approximately 1.0). Diltiazem (10(-5) M) lowers the KD for [3H]-nimodipine by a factor of 3 without changing the maximum number of binding sites. Diltiazem decreased the dissociation rate constant of the nimodipine receptor complex from 0.18 min-1 to 0.049 min-1 and altered the pharmacological profile as revealed by displacement studies with (-) and (+) verapamil, (-) and (+) prenylamine and 1,4 dihydropyridines. In conclusion [3H]-nimodipine binding can be utilized as a tool to evaluate complex molecular interactions between calcium channel blockers. PMID- 6292745 TI - [125I] N6-p-Hydroxyphenylisopropyladenosine, a new ligand for Ri adenosine receptors. AB - N6-p-Hydroxyphenylisopropyladenosine (HPIA) has been labelled with carrier-free Na[125I] to very high specific activity (2,175 Ci/mmol) and used as an agonist ligand to characterize Ri adenosine receptors in rat cerebral cortex membranes. The binding is saturable, reversible, stereospecific and dependent on protein concentration. The specific binding at 37 degrees C was of high affinity with an equilibrium dissociation constant KD of 0.48 nmol/l and was saturable with 0.23 pmol of [125I]HPIA per mg of protein. The rate constant of association, k1, was 3.25 x 10(8) l mol-1 min-1 and that of dissociation, k2 0.0110 min-1 yielding at t1/2 of 63 min. In competition experiments the (-)isomer of N6 phenylisopropyladenosine (PIA) was 16-fold more potent than the (+)isomer in competing for binding sites. Specific binding was most effectively displaced by N6-cyclohexyladenosine (CHA, ki=0.26 nmol/l), (-)PIA (ki= 0.33 nmol/l) and HPIA (ki=0.52 nmol/l), whereas 5'-N-ethylcarboxamidoadenosine (NECA, ki=1.42 nmol/l) was less effective. The methylxanthines 3-isobutyl-1-methylxanthine (IBMX), theophylline and caffeine which have been classified as adenosine antagonists had ki values between 5-43 mumol/l. Binding of [125I]HPIA was regulated by guanine nucleotides and divalent cations. The results indicate that [125I]HPIA labels Ri adenosine receptors in rat brain membranes. PMID- 6292746 TI - [Diagnosis, epidemiologic aspects and prevention of hepatitis A]. PMID- 6292747 TI - [Local anesthetics, basic mechanism of action and clinical use]. PMID- 6292748 TI - [Monovalent cation selectivity of EDTA-modified calcium channels]. AB - Properties of slow inward sodium currents evoked by membrane depolarization in calcium-free EDTA-containing extracellular solution were studied on isolated intracellularly dialyzed neurons of the mollusc Helix pomatia. The relative permeability of the corresponding ionic channels determined from the shifts of the reversal potential was PNa+ :PLi+ :PNH2NH3+ :POHNH3+ = 1.00:0.80:0.55:0.21. For original sodium channels of the same membrane the relative values were 1.00:1.04:0.44:0.19. The induced sodium current could be blocked by calcium channels blockers D-600 and niphedipine more effectively than calcium currents in the same membrane (the corresponding dissociation constants were 10(-5)M and 0.8 . 10(-5)M for the induced sodium current comparing with 2.6 . 10(-5)M and 2.3 . 10(-5)M for the calcium current. On the basis of these data a suggestion is made that the calcium channels possess a main selectivity filter which is similar to that of sodium channels, and an additional external binding site which binds divalent cations and prevents monovalent cations from entering the channel. Calcium-chelating agents introduced into calcium-free extracellular solution release this binding site from the occupying ion and thus transform the calcium channel into a sodium one. PMID- 6292749 TI - [Hydrogen ion currents through sodium channels in myelinated nerve fiber membranes]. AB - Ionic currents in the nodal membrane of myelinated frog nerve fibre were measured under voltage clamp conditions when the Ranvier node was bathed in solutions containing impermeant cations instead Na. At pH lower than 4.0 small (less than 0.1 nA) currents were detected which rose to peak and then decayed more slowly. Kinetics and voltage range of activation of these currents were similar to those of usual sodium currents at low pH. These currents were reversibly blocked by benzocaine (1 mM). All this permitted identifying them as currents through sodium channels. Experiments in which concentrations of substituting cations (tris+, choline+), Ca2+ and H+ ions were varied showed that the inward currents observed are carried by hydrogen (or hydronium) ions. According to reversal potential measurements the relative permeability of the channels (PH/PNa) is equal to 203 +/- 14 on the average. It is concluded that the energy barriers for H+ in sodium channel are much lower than for Na+, but their passage through the channel is slow because of binding to an acidic group in the channel. PMID- 6292750 TI - [Hydrogen currents through aconitine-modified sodium channels in nerve fiber membranes]. AB - Ionic currents in nodal membrane treated with aconitine were measured under voltage clamp conditions when nodes were bathed in Na-free solutions. At pH lower than 4.6 inward ionic currents were detected which had kinetics and voltage range of activation analogous to those of aconitine-modified sodium channels at low pH. These currents were blocked by benzocaine (2 mM). Experiments with various concentrations of Ca2+, tris+, TEA+, choline+ ions showed that these ions are essentially impermeable both at normal and acidic pH. It is concluded that the inward currents observed are carried by H+ (or H3O+) ions through aconitine modified sodium channels. From reversal potential measurements relative permeability (PH/PNa) of sodium channels is estimated to be 1059 +/- 88. The results suggest that the aconitine-modified channel is a rather wide water-filled pore and the rate of H+ passing through the channel is limited by its binding to an acidic group. PMID- 6292751 TI - [Inactivation of calcium channels in the somatic membrane of mollusk neurons]. AB - The time-dependent decrease of calcium inward currents (inactivation) at long (about 1s) depolarizing membrane potential shifts was studied on isolated dialysed neurons of the mollusc Helix pomatia. It is shown that the time course of this decrease can be approximated by two exponents with corresponding time constants 20-70 and 250-350 ms. The fast component of the decay disappeared completely after pH increase of the intracellular solution to 8.5. The kinetics of the slow component did not change. A conclusion is made that the fast decay of the recorded current does not represent variations in the calcium current but is due to parallel activation of a nonspecific outward current whereas the slow decay represents true inactivation of the calcium current. It is shown that the inactivation velocity of this current is related to the maximal value of the latter but not to the level of membrane depolarization, and depends on conditions of accumulation of calcium ions at the inner surface of the cell membrane. PMID- 6292752 TI - [Activation of motoneurons of the nucleus of the accessory nerve by rubrofugal impulses in the cat]. PMID- 6292753 TI - [Depolarization of primary afferents in the spinal cord. Mechanisms and functions]. PMID- 6292754 TI - [The cerebral refractory period of the somatosensory system in the diagnosis of atrophic brain diseases]. PMID- 6292755 TI - [Hemorrhage in hypophyseal tumors and in the chiasm area--urgent surgical indications]. PMID- 6292756 TI - [Familial recurring polytopic neuropathy. A case report]. PMID- 6292757 TI - [Does 24,25 dihydroxycholecalciferol have a physiological and pathophysiological role?]. AB - The authors review recent experimental and human data concerning the potential physiological and pathophysiological role of 24,25 (OH)2D3, the dihydroxylated metabolite of vitamin D which is synthetisized with preference over 1,25 (OH)2D3 in organisms that have been replenished with vitamin D. For the major known effects of vitamin D such as stimulation of intestinal absorption and bone resorption of calcium and phosphorus, 24,25 (OH)2D3 is less effective than the 1,25 (OH)2D3 metabolite and consequently of lesser physiological importance. Some recent in vitro experiments have shown, however, that 24,25 (OH)2D3 intervenes in the stimulation of proteoglycan synthesis, inhibition of PTH, vitamin A and heparin induced resorption, whereas 1,25 (OH)2D3 does not. Although there is controversy as to its direct inhibitory effect on secretion of PTH, it seems to act with 1,25 (OH)2D3 to prevent hyperplasia of parathyroids in vitamin D deficient chicken. From a pathophysiological point, the presence of 24,25 (OH)2D3 seems vital to allow normal bone formation and mineralisation and possibly to counteract excessive bone resorption. PMID- 6292759 TI - [Evaluation of sulocton action in patients with atherosclerotic brain ischemia]. AB - Sulocton effect was evaluated in 33 patients with symptoms of brain ischaemia during atherosclerosis. Thirty patients received the drug orally for 2 months in doses of 100 mg thrice daily. Three patients discontinued the treatment earlier (two of them discontinued it because of side effects). After two months of treatment a significant improvement was observed in such disturbances important for wellbeing and social contacts as: anxiety and fear, mood depression, disequilibrium of emotion, motivation and initiative. In many patients headaches, dizziness and gait disturbances disappeared or diminished. Sulocton was useful in the treatment of patients with cerebral atherosclerosis. PMID- 6292758 TI - Isethionate and taurine transport sites at brain cell membranes: influx studies with brain slices. PMID- 6292760 TI - [Treatment of acromegaly by the surgical approach through the sphenoid bone. I. Clinical results]. AB - Forty-five cases of pituitary adenomas with clinical characteristics of acromegaly treated surgically from an approach through the nose and sphenoid sinus are presented. After the operation the somatic features of acromegaly decreased in 66% of cases, headaches regressed in 88%, visual field defects disappeared in 20% of cases. Selective removal of pituitary microadenoma caused return of regular menstrual cycles in 12 women and in 3 of them successful pregnancy and labour resulted. In cases of extrasellar growth of the tumour no satisfactory clinical remission was obtained. PMID- 6292761 TI - Foot shock analgesia. Lack of correlation with pituitary and plasma immunoreactive-beta-endorphin. AB - Effects of inescapable foot shock on pain threshold and on levels of immunoreactive beta-endorphin (ir-beta EP) in anterior pituitary, neuro intermediate lobe and plasma were determined by hot plate test and radioimmunoassay respectively. Whereas handling and conditioned stress failed to alter baseline pain threshold, 20 min inescapable foot shock produced modest and transient analgesia. Levels of plasma ir-beta EP and corticosterone were significantly and concurrently raised by all treatments and remained elevated 40 min after foot shock. Changes in plasma levels of ir-beta EP and corticosterone were found to parallel one another but did not correlate with changes in pain threshold. AP content of ir-beta EP was significantly reduced 60 min after the cessation of inescapable foot shock, and N-IL content from 20 min after. The findings suggest that (i) plasma levels of ir-beta EP and corticosterone do not reflect changes in pain threshold, and (ii) ir-beta EP levels in anterior pituitary, neurointermediate lobe and plasma probably are not causally related to foot shock analgesia. PMID- 6292762 TI - Comparative study in vivo and in vitro of the differentiation of immunoreactive corticotropic cells in fetal rat anterior pituitary. AB - In order to study the mechanisms of the differentiation of adenohypophysial corticotropic cells, an immuno-cytological study was performed in fetal rat anterior pituitary in vivo and in vitro with antisera against beta-(1-24) and alpha-(17-39) ACTH and beta-LPH, alpha- and beta-endorphins. In vivo, these cells appeared at 16 days of gestation without any difference in the timing of appearance of the two immunoreactivities. The same immunoreactivity was also detected in adenohypophysial primordia explanted from 12 to 15 days of gestation and maintained in organ culture until 21 days by using either medium containing fetal calf serum or medium containing insulin and transferrin instead of fetal calf serum. These immunoreactive cells were first detected in the different experimental primordia after a minimal period of culture, corresponding to a final equivalent of 16 days as in vivo. However, the mean cytoplasmic area of immunoreactive cells increased in relation to the day of explantation whatever the duration of culture. These data suggest: (1) the nature of culture medium used in this study has no influence on the differentiation of the corticotropic cells; (2) this cell type seems to be committed precociously (before day 12) by one or several substances of unknown origin; (3) the normal development seems to require the presence of factors (before day 14) whose nature and origin remain to be elucidated. PMID- 6292763 TI - Adrenocorticotropic regulations after bilateral lesions of the paraventricular or supraoptic nuclei and in Brattleboro rats. PMID- 6292765 TI - Stability of pro-opiocortin-related peptides in post-mortem mouse brain tissue. AB - The stability of pro-opiocortin-related peptides, growth hormone and somatostatin was investigated in mouse brain for up to 72 h post mortem. The peptide content in acid extracts of whole mouse brain was measured by radioimmunoassay and the molecular forms characterised by chromatography on Sephadex G-50 under acid dissociating conditions. The brain content of the pro-opiocortin-related peptides and growth hormone rose markedly with time after death and chromatography showed this to be due to an increase in the same molecular forms present immediately post mortem, with no significant peptide fragmentation or precursor cleavage. These rises were not seen in the brains of those animals hypophysectomized at death. In contrast, the levels of somatostatin were not significantly altered post mortem though there was a shift in the relative distribution of immunoactivity between the different molecular forms. These results indicate that the peptide content of post-mortem mouse brain may be affected by leakage of pituitary contents and that even though the levels of other neuropeptides may not change, their molecular forms may be altered. PMID- 6292764 TI - Prolactin and osmoregulation in vertebrates. An update. PMID- 6292766 TI - Peripheral versus central cardiorespiratory effects of morphine. AB - The administration of morphine sulfate (MS, 2 mg/kg) into the right atrium in decerebrate rats, produced a dramatic bradycardia, apnea, and a slight transient biphasic blood pressure response within 1 sec. The bradycardia was slowly restored to control levels within 8-10 min and was prevented by pretreatment with atropine. Apnea (7.3 +/- 2.2 sec) was followed by a period of rapid shallow breathing. Acute tolerance to these effects developed to subsequent doses of morphine. In paralyzed artificially ventilated animals, morphine produced: (1) cessation of phrenic nerve (PN) activity which was followed by bursts of shortened duration; and (2) concomitant excitation of the recurrent laryngeal nerve (RLN), this activity exhibited a continuous discharge which was asynchronous with that of phrenic nerve. These actions of morphine were qualitatively similar to those observed with phenyldiguanide (PDG) and were found to be reflexogenic, i.e. emanating from the lungs via stimulation of pulmonary C fibers. Bilaterally vagotomized animals only responded to morphine with respiratory depression, characterized by a severe decrease in respiratory rate and minute ventilation. On the other hand, animals with intact vagi showed stimulation of rate and minute ventilation following a brief period of apnea. They were less sensitive than vagotomized animals to the depressant action during the first 8 min after administration of morphine. All the effects of morphine were blocked by pretreatment with naloxone (50-400 micrograms/kg, i.v.). These results indicate that the initial cardiorespiratory effects of morphine are due to a peripheral reflex action arising from the stimulation of opiate receptors associated with pulmonary C-fibers, e.g. J-fibers. PMID- 6292767 TI - Raphe unit activity in freely moving cats: effects of benzodiazepines. AB - Benzodiazepines (chlordiazepoxide and diazepam) produced a dose-dependent decrease in the discharge rate of serotonin-containing neurons in the dorsal raphe nucleus of freely moving cats. This ranged from no significant change at doses of 0.5 and 1.0 mg/kg (i.p.), to greater than 90% reductions in unit activity at 10 mg/kg. The effects of benzodiazepines on raphe units occurred within 15-30 min of injection and the duration of action was dose-dependent and lasted from 1 to more than 6 hr. Doses of benzodiazepines that significantly decreased raphe unit activity (i.e. 2.5-10 mg/kg) also produced ataxia and decreased EMG activity. These data suggest that benzodiazepine-induced suppression of raphe unit activity is closely related to general motor behavior. Raphe unit activity remained suppressed during phasic increases in EMG activity during eating, grooming, or predatory behavior, suggesting that benzodiazepines also have a direct inhibitory action on raphe cells. The present results are discussed in the context of the serotonergic hypothesis of anxiety. PMID- 6292769 TI - GABAergic regulation of the substantia innominata-cortical cholinergic pathway. AB - Neurochemical and immunohistocytochemical studies have demonstrated a nucleus accumbens-substantia innominata GABAergic pathway. The present authors' data with muscimol given parenterally, as well as local injections into the substantia innominata, further support an inhibitory function for this pathway in the regulation of the substantia innominata-cortical cholinergic projection. PMID- 6292768 TI - Ethanol and acetaldehyde alter brain mitochondrial redox responses to direct cortical stimulation in vivo. AB - To determine whether and how ethanol and acetaldehyde alter brain oxidative metabolic activity, reduction/oxidation shifts of components of the mitochondrial respiratory chain were optically measured, in situ, from cat cerebral cortex. Oxidative shifts of nicotinamide adenine dinucleotide (NADH) were recorded in response to increased energy demand provoked by stimulation of the cortical surface by electrical pulses. Ethanol or acetaldehyde did not alter the direction of the responses but each slowed the rates of oxidation with little effect upon the rates of subsequent re-reduction. There was no apparent change produced by either drug upon the kinetics of the negative shifts of the cortical steady potential in response to the stimulation. However, stimulus-evoked electrical and metabolic responses were decreased in amplitude with increasing drug doses. It is suggested that the slowed mitochondrial oxidation results from inhibition of Na+, K+-ATPase. This supports the concept that ethanol or acetaldehyde inhibit the processes that lead to increased oxygen consumption following cell depolarization in vivo, as has been demonstrated in vitro. PMID- 6292770 TI - The effects of pentylenetetrazol, bicuculline and strychnine on the development of kindled seizures. PMID- 6292771 TI - Convulsant and anticonvulsant effects of opioids: relationship to GABA-mediated transmission. AB - The convulsant benzodiazepine Ro 5-3663, bicuculline and picrotoxin induced electroencephalographic (EEG) and behavioural convulsions. In rabbits, the EEG modifications consisted, with increasing doses, of three different patterns: slow waves in the optic lead, spike- and wave-complexes in the sensorimotor cortex, and grand-mal generalized seizures. These EEG effects were terminated by administration of diazepam (1 mg/kg) and morphine (0.25-1.0 mg/kg). Naloxone, in doses of 5-10 mg/kg, potentiated the effects of the three convulsant drugs. This potentiating phenomenon was also antagonized by the administration of diazepam and morphine. In membrane preparations, obtained from rat cortex, deprived of endogenous modulators of [3H]GABA binding, naloxone but not morphine, was able to inhibit [3H]GABA binding to its specific recognition sites. These data agree with previous findings indicating a GABA-antagonistic effect of naloxone, and support the hypothesis that the anticonvulsant effect of morphine might be, at least in part, due to an increase in GABAergic activity at the synaptic level. PMID- 6292772 TI - Comparative effects of the ACTH 4-9 analogue (ORG 2766), ACTH 4-10 and [D-Phe7] ACTH 4-10 on medial septal self-stimulation behaviour in rats. AB - Experiments were performed to examine the effects of various analogues of ACTH on electrical self-stimulation behaviour elicited from the medial septal area using an ascending or descending sequence of stimulus intensities within a session. When an ascending sequence of threshold multiples was used ACTH 4-10 and the ACTH 4-9 analogue (ORG 2766) enhanced level pressing for low intensity stimulation but attenuated self-stimulation at greater current intensities. The analogue ORG 2766 appeared to be a thousand times more potent than ACTH 4-10; [D-Phe7] ACTH 4-10 inhibited the response rate at threshold level but was inactive at greater current intensities. The same effect was found following administration of ORG 2766 in a dose which was 20 times greater (1 microgram/rat) than that used in the first experiments. Lever pressing was not affected by treatment with ACTH 4-10 or ORG 2766 when a descending sequence of stimulus intensities was used within a session. Thus, ACTH-related peptides may affect motivational processes involved in self-stimulation rather than the reward of the stimulation per se. It is suggested that although ORG 2766 mimicked the action of ACTH 4-10 this synthetic peptide may have additional behavioural properties. PMID- 6292773 TI - Effect of opiate receptor agonists on striatally-mediated head turning: an in vivo model of opiate delta receptor activation? AB - Electrical stimulation of the striatum in conscious rats elicits a contralateral head-turn. Modification of this response by opiate drugs injected sub-cutaneously or into the ipsilateral globus pallidus has been investigated. Morphine, normorphine and ethylketocyclazocine had no effect on the head-turn latency but met-enkephalin and D-Ala2, D-Leu5-enkephalin, injected into the pallidum, and smalL doses of etorphine injected subcutaneously, produced a dose-related slowing of the response. Large doses of the opiate receptor antagonist naloxone antagonised the effect of etorphine. These results suggest the presence of opiate delta receptors in the rat globus pallidus and this model may therefore prove useful for studying the effects of other agents acting at the delta receptor. PMID- 6292774 TI - The time course and specificity of the changes in the behavioural and dorsal horn cell responses to noxious stimuli following peripheral nerve capsaicin treatment in the rat. AB - Capsaicin, a neurotoxin which acts specifically on sensory primary afferent C fibres was applied locally to one sciatic nerve of a group of rats. One to sixteen days following this a series of behavioural and electrophysiological studies were performed. The latency of foot withdrawal of the rats to a controlled thermal noxious stimuli was significantly elevated (200%). The peak increase occurred on day 1 after treatment; the response then fell to a steady but elevated level for up to 16 days. Responses to noxious mechanical stimuli were unaffected by capsaicin treatment. Single unit analysis of the dorsal horn of the spinal cord showed that the number of neurones in deep laminae (4, 5 and 6) responding to a C peripheral volley was normal (60%) for the first 2 days after treatment. On day 3 post treatment, the number of cells with a C input began to fall reaching a maximal decrease on day 7 (25%), where it remained up to day 16. In contrast to this delayed effect on C-evoked responses, the number of cells responding to noxious heating of the skin fell from control levels of 60% down to 20% on day 1 and remained decreased for up to 16 days. The onset of thermal analgesia following local, capsaicin treatment, therefore, closely parallels the time course of the decrease of noxious heat-evoked responses in the dorsal horn. Since at early pretreatment times, the electrically C-evoked activity is normal these effects are likely to be due to action on peripheral C fibre nociceptors in the skin. At a later stage capsaicin also appears to act on the central terminals of fibres reducing transmission to second order dorsal horn neurones. PMID- 6292775 TI - Separation of temperature sensitive and temperature insensitive components of the postsynaptic potentials in the frog motoneurons. AB - Intracellular measurements were made in the in situ spinal cord of the frog at temperatures below 5 degrees C. Responses to volleys in the sciatic nerve, in the descending fibres and in the motor axons were studied. About 30% of the motoneurons responded to sciatic volleys with 1-3 ms segmental latency, which was short enough to assume electrotonic mediation of these responses. Another group of motoneurons responded with 6-8 ms latency, i.e. with the expected delay at chemical synapses at low temperature. Latency distribution of the sciatic-evoked postsynaptic potentials was clearly bimodal in contrast with that found at higher temperatures. Postsynaptic discharges occurred with rather long latency and they were attributed to chemically-mediated excitation. Some of the postsynaptic potentials to descending volleys also occurred with quite short latency, indicating possible electrotonic transmission from supraspinal centres to motoneurons. Latency distribution of the action potentials evoked from the motor axons was bimodal, corresponding to the different, i.e. antidromic and recurrent facilitatory, mechanism of these spikes. Calculated Q10 ratios for the sciatic evoked reflex discharges and the afferent fibre volleys were about 2.3 and 1.8, respectively. We concluded that cooling helps to separate postsynaptic potentials according to their electrotonic and chemical mediation and that electrotonic excitation does not seem to have a primary role in the generation of postsynaptic discharges initiated by dorsal root volleys in the frog. PMID- 6292776 TI - Presynaptic acetylcholine receptors at the excitatory amino acid synapse in locust muscle. AB - Acetylcholine (greater than or equal to 10(-5) M) applied in the bathing medium to the excitatory nerve-muscle junction of the locust caused a dose-dependent increase in the frequency of spontaneous miniature potentials and in the mean quantal content of evoked potentials. The statistical characteristics of the spontaneous release process were also modified, high frequency bursts of spontaneous potentials and "giant' miniature potentials occurring in the presence of acetylcholine. The response, which was dependent on extracellular calcium concentration, consisted of two distinct phases; these could be induced or abolished selectively by nicotinic and muscarinic drugs, respectively. The results suggest the presence, in low density, of two types of cholinergic receptors on the excitatory motor nerve terminals of the locust. The acetylcholine-induced channels may admit calcium ions. The possible role of these presynaptic receptors is discussed. PMID- 6292777 TI - Intracellular metabolism of adenosine 3',5'-cyclic monophosphate and calcium inward current in perfused neurones of Helix pomatia. PMID- 6292778 TI - A reanalysis of the ventrolateral input in slow and fast pyramidal tract neurons of the cat motor cortex. AB - In deeply anesthetized cats the temporal characteristics of ventro-lateral (thalamic) excitatory postsynaptic potentials (EPSPs) induced in pyramidal tract cells were studied with an averaging technique. Stimulation of the ventrolateral thalamus induced EPSPs in all pyramidal tract neurons at latencies of 1-5 ms. It was found that there was a positive relationship between the latency and rise time of stimulation-induced EPSPs and the latency of antidromic invasions of pyramidal tract neurons. In response to two closely spaced shocks the second EPSP had the same latency and amplitude as the first one in both slow and fast pyramidal tract neurons. Moreover, the span of antidromic latencies of ventrolateral thalamic relay cells to motor cortex stimulation showed that these thalamic neurons had the necessary conduction velocities to account for the distribution EPSP latencies recorded in pyramidal tract neurons. From these electrophysiological results, it has been concluded that slow and fast pyramidal tract morphological evidence, obtained at the electron-microscopic level, supporting this conclusion. Terminal degeneration induced by a lesion in the ventrolateral thalamus was found on the apical dendrite of a slow pyramidal tract neurons that had been injected with horseradish peroxidase. It is proposed that the matching between the latencies of EPSPs evoked from the ventrolateral thalamus and the latencies of antidromic invasions of pyramidal tract neurons may reflect a matching between the conduction velocity of thalamocortical and cortico spinal neurons and/or it may be due to the electrotonic properties of the apical dendrites. PMID- 6292779 TI - Electrophysiological and pharmacological characteristics of facilitation of hippocampal population spikes by stimulation of the medial septum. AB - In rats under urethane anesthesia, single shock or tetanic stimulation of the medial septum--which evoked only minimal field potentials--sharply enhanced population spikes evoked in area CA1 by commissural stimulation. An enhancement of population spikes was observed only (a) in areas CA1 and CA2 (adjacent to CA1 in the dorsal hippocampus), but not in the fascia dentata or the deep pyramidal layers CA3 or CA4; (b) in a narrow range of depth, close to the stratum pyramidale; (c) when the intensity of commissural stimulation was of adequate intensity. A comparable facilitation of population spikes was produced at the same sites by microiontophoretic release of acetylcholine. The septal facilitatory action increased in effectiveness with the number of tetanic pulses (up to 10-12) at a given frequency, and it had a maximum at frequencies of 50-100 Hz. It reached a maximum 20-50 ms after the end of septal stimulation, and then decayed slowly, the overall duration being up to 300 ms. The cholinergic nature of the facilitation induced by septal stimulation was confirmed by the parallel potentiation of septal action and that of acetylcholine by physostigmine and their depression by atropine and scopolamine. PMID- 6292780 TI - Responses of supraoptic neurons to electrical stimulation of the medial amygdaloid nucleus. PMID- 6292781 TI - Changes in noradrenergic terminal excitability induced by amphetamine and their relation to impulse traffic. AB - The effects of amphetamine upon the terminal excitability of noradrenergic neurons of the nucleus locus coeruleus were studied in urethane anesthetized rats. Terminal excitability was measured by determining the stimulus currents necessary to evoke antidromic responses in locus coeruleus neurons from terminals in the frontal cortex. In most cases, terminal excitability was decreased following local infusion of amphetamine into the frontal cortex, while intravenous administration of the drug tended to increase terminal excitability. The decreased terminal excitability induced by local infusion of amphetamine appeared to be due to activation of alpha-adrenergic receptors located on the terminals of locus coeruleus neurons, since this effect mimics that of clonidine, a direct acting alpha-adrenergic agonist, and since the effect was abolished by pretreatment with alpha-methyl-p-tyrosine which disrupts the catecholamine liberating properties of amphetamine. Phentolamine, a direct acting alpha adrenergic receptor antagonist was also found to block or reverse the effect of amphetamine. The changes in terminal excitability following intravenous injection of amphetamine appeared to be related to changes in the spontaneous activity of locus coeruleus neurons. A large decrease in spontaneous activity following intravenous administration of amphetamine was associated with increased terminal excitability, whereas when smaller changes in spontaneous activity occurred, terminal excitability was found to be decreased. These results are discussed with respect to the pharmacological properties of catecholaminergic neurons and the mechanisms of action of amphetamine. PMID- 6292782 TI - Electrotonic synapses are formed by fetal rat sympathetic neurons maintained in a chemically-defined culture medium. AB - Principal neurons from the superior cervical ganglia of rat fetuses were maintained for up to 101 days in dissociated cell cultures in a serum-free, chemically-defined medium; non-neuronal cells were killed by the periodic addition of fluorodeoxyuridine to the medium. Intracellular recordings, obtained at various times between 16th and 98th day in vitro, showed that these neurons could generate substantial (up to 90 mV) action potentials in response to depolarizing current injections; these responses were dependent on tetrodotoxin sensitive Na+ channels, cobalt-sensitive Ca++ channels, and tetraethylammonium sensitive K+ channels. Action potentials were often followed by prominent, long hyperpolarizing after-potentials (10-15 mV, greater than 150 ms); the duration of these after-potentials was reduced by the addition of Co++ (2-5 mM) to the perfusate. Acetylcholine depolarized these neurons by a hexamethonium-sensitive mechanism. To determine whether sympathetic neurons formed synapses in a defined medium, intracellular recordings were obtained from pairs of neighboring neurons. Synaptic interactions were frequently observed at all times in vitro (up to 60% of all pairs tested). At many synapses, both hyperpolarizing and depolarizing DC potential changes spread from one neuron to another. At other synapses, the spread of DC potential changes could not be directly demonstrated; however, interactions at such synapses were not inhibited by antagonists of several neurotransmitters, by elevation of the Mg++/Ca++ ratio, or by the addition of Co++. Thus most, if not all, of the synaptic interactions among sympathetic neuron were electronic; such electrical synapses were not observed among dorsal root ganglion neurons maintained in the same medium. These data indicate that, when maintained in a chemically-defined culture medium, sympathetic neurons of rat fetuses express many of the basic membrane properties observed in neurons of superior cervical ganglia recently removed from adult rats. However, fetal sympathetic neurons maintained in this defined medium also differ from their counterparts in vivo; they adopt a mode of synaptic transmission (electrical) that has not been observed in the sympathetic ganglia of the adult rat. Thus, as late as the 21st embryonic day, not only the choice of neurotransmitter, but also the mode of transmission has not been irrevocably determined in sympathetic neurons. PMID- 6292783 TI - Release of [3H]noradrenaline from descending tracts in the cat spinal cord in vivo. AB - The release of [3H]noradrenaline into the perfused central canal of the cat lumbar-sacral spinal cord was monitored in vivo. Stimulation of descending tracts produced an increase efflux into artificial cerebrospinal fluid containing 10(-6) M phenoxybenzamine which could be dissociated from any concurrent rise in blood pressure. No release was produced by stimulating dorsal roots over the length of cord perfused. It appears, therefore, that noradrenaline can be released from descending nerve terminals, but not from dorsal root afferents in the spinal cord. PMID- 6292784 TI - Mechanism of presynaptic actions of adenosine and acetylcholine on noradrenaline release in the guinea-pig heart. PMID- 6292785 TI - Trifluoperazine, a calmodulin inhibitor, blocks secretion in cultured chromaffin cells at a step distal from calcium entry. AB - Trifluoperazine, a calmodulin antagonist, inhibited the secretory response of cultured bovine adrenal medullary chromaffin cells to acetylcholine (10(-4) M) or a depolarizing concentration of [K+] (56 mM KCl) in a dose-related fashion. The ID50s of this effect were 2 x 10(-7) M and 2.2 x 10(-6) M for acetylcholine and high [K+], respectively. A decrease in external [Ca2+] concentration of the incubation medium from 4.4 to 0.275 mM resulted in an increase in the percentage of inhibition produced by trifluoperazine on the acetylcholine-evoked secretory response from 20.7 to 96.5%, respectively. However, trifluoperazine inhibited the acetylcholine-evoked catecholamine output by a similar absolute magnitude for all [Ca2+] concentrations tested with the exception of 4.4 mM [Ca2+]. Trifluoperazine, unlike the [Ca2+] channel blocker Ni2+, in concentrations (10( 6)-10(-5) M) that were found to inhibit significantly [K+]-induced amine output did not modify [K+]-induced 45Ca uptake or 45Ca efflux. However, trifluoperazine at a concentration of 2.5 x 10(-5) M was found to produce a small decrease in the 45Ca efflux curve and a decrease in the [K+]-evoked 45Ca uptake of 30 +/- 14% (n = 6). In addition, 2.5 x 10(-6) M trifluoperazine, a concentration which was found to suppress high [K+]-induced amine release by 64 +/- 5%, did not inhibit the 45Ca2+-Ca2+ exchange mechanism. These results demonstrate that trifluoperazine, an antipsychotic agent with anticalmodulin activity, blocks catecholamine release from cultured chromaffin cells at a step distal from calcium entry and, consequently, suggests a role for calmodulin in the secretory process of these cells. PMID- 6292786 TI - Attenuation of induced-anxiety in rats by chlordiazepoxide: role of raphe dorsalis benzodiazepine binding sites and serotoninergic neurons. AB - In chronically implanted awake rats, microinjections of chlordiazepoxide (5 x 10( 7) M) into the dorsal raphe significantly attenuated the inhibition of lever pressing for food elicited by a signal of punishment. This effect is abolished by prior application of 5,7-dihydroxytryptamine into the dorsal raphe (3 weeks after the infusion of the neurotoxin, dorsal raphe tryptophan hydroxylase activity was reduced to 25% of control values). Furthermore, the disinhibitory effect of intra raphe chlordiazepoxide can be mimicked or potentiated by intra raphe dorsalis application of serotonin (10(-7) or 10(-8) M, respectively). Further evidence for a crucial interaction between benzodiazepines and serotoninergic processes are provided by in vitro experiments showing that chlordiazepoxide or diazepam (10( 5) M) are able to facilitate the K+-evoked [3H]serotonin release from rat midbrain slices. Finally, a high density of [3H]flunitrazepam binding sites was found in the dorsal (and the median) raphe nucleus, the Kd and Bmax values being not altered by prior infusion of 5,7-dihydroxytryptamine. These in vitro data suggest possible means by which intra raphe (and perhaps peripherally administered) benzodiazepines may affect the activity of serotoninergic neurons and thereby produce their effects on experimental anxiety. PMID- 6292787 TI - Somatosensory evoked potentials in patients with supraclavicular brachial plexus injuries. AB - In 12 patients with traumatic lesions of brachial plexus, we studied somatosensory evoked potentials by stimulation of median, radial, and ulnar nerves at the wrist and by recording at the arm, Erb point, cervical spinal cord, and contralateral cortex. Results after median nerve stimulation in patients with upper trunk lesions were normal. Patients with C5/6 root avulsion had either normal, delayed, or absent responses at the cervical cord and cortex, depending on the presence of complicating distal lesions. In patients with particular involvement of C7 root, results were normal after median and ulnar stimulation but were abnormal after radial nerve stimulation. In patients with multiple root avulsions and flail anesthetic arm, no potentials could be recorded from the cervical cord or contralateral cortex, regardless of which nerve was stimulated. For relevant information, it was important to stimulate nerves having roots near the anatomic site of the lesion as determined clinically and electromyographically. PMID- 6292788 TI - Characterization of beta-adrenergic receptors on human cerebral microvessels. AB - Cerebral microvessels were separated and prepared from human brain cortex by albumin floatation and glass bead filtration. The binding of a specific beta adrenergic antagonist, (125I) iodohydroxybenzylpindolol, to the microvessel preparation was characterized by high affinity, saturability, and stereospecificity. The presence of specific beta-adrenergic receptor sites in human cerebral microvessels supports the hypothesis of adrenergic regulation of the function of cerebral microvessels. PMID- 6292790 TI - Electron diffraction of thin-sectioned biological specimens--an efficient but neglected method for lattice assessment. PMID- 6292789 TI - Quantitative autoradiography of neurotransmitter receptors in Huntington disease. AB - We studied gamma-aminobutyric acid (GABA), benzodiazepine, and muscarinic cholinergic receptor-binding by quantitative autoradiography. In coronal sections from the brain of a patient with Huntington disease, binding for all three receptors in caudate and putamen was lower than control values. Binding to GABA and benzodiazepine receptors was increased in lateral and medial pallidum and decreased in ventrolateral thalamus. Muscarinic cholinergic receptors were markedly decreased in pallidum but not thalamus. The findings suggest that loss of striatal afferents to both segments of pallidum results in GABA and benzodiazepine receptor supersensitivity, and support the utility of quantitative autoradiography for receptor studies in human postmortem material. PMID- 6292791 TI - [Ovarian thecoma. Description of 2 cases]. PMID- 6292792 TI - [Effect of salbutamol, indomethacin and atenolol on plasma renin activity in patients with essential arterial hypertension]. PMID- 6292793 TI - Single potassium channel currents in cultured mouse oligodendrocytes. AB - Single channel currents were recorded from isolated membrane patches of cultured mouse oligodendrocytes using the giga-seal technique. The observed conductance of the channel was 71 +/- 34 pS. Isolated patches contained 1 to 4 channels with similar conductances and kinetics. Closed times of the channel varied from less than 1 msec to many minutes. The open state was always interrupted by flickering to the closed state. The kinetics of opening and closing appeared insensitive to voltage steps of up to +/- 75 mV from the resting level of the membrane potential, but could be affected by very large voltage steps. The observed changes in channel current in response to changes of potassium concentration on either side of the membrane indicate a high selectivity for potassium. The results show a membrane with constant macroscopic permeability that contains channels which open and close. PMID- 6292794 TI - Lack of alpha-adrenoreceptor binding of [125I]BE2254 to rat basilar artery membranes. AB - Rat basilar arteries do not contain classical alpha- or beta-adrenoreceptors as assessed by electrophysiological techniques even though these arteries are innervated by catecholamine-containing perivascular nerves. These arteries were therefore examined for their ability to selectively bind an alpha-adrenoceptor radioligand, [125I]BE2254 (2/beta/4-hydroxyphenyl)-ethylaminomethyl)-tetralone). For comparison, rat tail arteries were also studied as these are known to contain functional alpha-adrenoreceptors. It was found that basilar artery membranes had only one-third of the specific binding of tail artery membranes and this finding collaborates the electrophysiological data. PMID- 6292795 TI - Interactions of light and of tetraethylammonium-chloride (TEA) on action potentials in identified neurons of Helix pomatia. AB - In neurons B1, B2 and B3 of Helix pomatia and illumination elicited: (1) a reduction of the width of action potentials (AP) in control bath fluid; (2) an acceleration of the development of tetraethylammonium (TEA) action on AP; and (3) a diminution of the full established effect of TEA. The shortening of the AP by light was augmented with increasing prolongation of the AP, regardless of whether it was evoked by external or internal TEA or by depolarization of the cells. The effects may be interpreted on the basis of a single mechanism. PMID- 6292796 TI - Apparent delay between light-induced receptor current and receptor potential in the Limulus ventral nerve photoreceptor. AB - We measured receptor potential (ReP) and receptor current (ReC) under voltage clamp conditions alternatingly evoked by successive identical 10 ms light flashes in the ventral nerve photoreceptor of Limulus. At high stimulus intensities the apparent latency of the ReP is 4-15 ms shorter than that of the ReC. At lower light intensities the difference is even larger. High amplification reveals that the light response starts with two phases corresponding to two current components. The initial first current component rises linearly, is much smaller than the second one and therefore is not detected with standard amplification. This first component, which is more pronounced at low light stimulus intensities, is caused either by a displacement or by a weak ionic current. PMID- 6292797 TI - Intrinsic actions of benzodiazepine antagonists. AB - RO 15-1788 (4-20 mg/kg), both alone and in combination with chlordiazepoxide (5 mg/kg), increased exploratory head-dipping in an holeboard above control levels. This intrinsic action of a benzodiazepine antagonist is not reversed, and is even augmented, by chlordiazepoxide. RO 15-1788 alone had no effect on locomotor activity, but antagonized the sedative effects of chlordiazepoxide. beta-CCE (1 and 2 mg/kg) was without effect on locomotor activity, but the higher dose reduced head-dipping. PMID- 6292799 TI - Morphine catalepsy in the rat: involvement of mu 1 (high affinity) opioid binding sites. AB - Since the proposal of multiple classes of opiate receptors, much effort has been devoted to understanding the subtypes mediating specific opioid actions. Using the irreversible antagonist naloxazone, we have established a role for the high affinity (mu 1) sites in analgesia. These sites also appear important in opiate catalepsy. Control rats showed dramatic catalepsy following a high dose of morphine sulfate (100 mg/kg, s.c.). Rats whose high affinity (mu 1) sites had been blocked with naloxazone 24 h earlier, on the other hand, showed no evidence of catalepsy (P less than 0.01). Accordingly, catalepsy appears to involve the same receptors as analgesia, the higher affinity (mu 1) sites. PMID- 6292798 TI - Anisatin modulation of GABA- and pentobarbital-induced enhancement of diazepam binding in rat brain. AB - Anisatin, a pure toxic substance isolated from the seeds of a Japanese plant (Illicium anisatum) acts as a picrotoxin-like, non-competitive GABA antagonist. Anisatin inhibited [3H]diazepam binding enhanced by either GABA or pentobarbital, without affecting the basal specific binding to rat brain membranes. The inhibition of this pentobarbital enhancement was competitive. These actions of anisatin were even more apparent when the binding assays were carried out at 37 degrees C rather than a 0 degrees C. Thus, at a physiological temperature, anisatin may have a more potent modulatory effect on benzodiazepine-GABA receptor coupling, through the barbiturate-picrotoxin sensitive sites. PMID- 6292800 TI - cAMP-dependent and cAMP-independent modulation of synaptic transmission in guinea pig superior cervical ganglion. AB - RMI 12330A dose-dependently inhibits cAMP accumulation induced by PGE2 in guinea pig superior cervical ganglion (SCG). Norepinephrine (NE) is unable to modify cAMP concentration in the same preparation. On the other hand, both PGE2 and NE block the firing discharge of guinea-pig SCG neurons elicited by electrical stimulation (1 Hz) of the cervical sympathetic trunk. The effect of PGE2, but not that of NE, is prevented by RMI 12330A, suggesting that NE and PGE2 act through different pathways PMID- 6292801 TI - Myoglobinuria associated with parainfluenza type 2 infection. PMID- 6292802 TI - Renal oncocytoma. PMID- 6292803 TI - Single dose doxycycline, cefuroxime and pivmecillinam for treatment of bacterial cystitis. AB - There is now considerable evidence showing the benefits of single dose antibacterial treatment for uncomplicated urinary tract infections. If single dose therapy is to become widely used it is necessary to clarify the minimum effective dose of the most efficient drugs. This paper reports three trials in women presenting in general practice with bacterial cystitis. In each trial the patients were randomly allocated to either a five day course of oral co trimoxazole (CTM) or to doxycycline 300 mg orally, cefuroxime 1.5 g intramuscularly or pivmecillinam 600 mg orally. Thirty-eight of 45 patients treated with doxycycline were cured, compared with 44 or 45 treated with CTM. Fourteen of 20 women given cefuroxime were cured, compared with 19 of 20 prescribed CTM. Twenty-three of 30 women treated with pivmecillinam were cured, compared with all 30 given CTM. None of these three drugs, when administered as a single dose, was as effective as a single 1.92 g or 2.88 g dose of CTM used in our previous studies in domiciliary practice. These studies confirmed, however, that single dose therapy was well tolerated, preferred by the patients and side effects were minimal. PMID- 6292804 TI - [A course study on schizophrenic psychoses within the scope of psychiatric before and after care]. PMID- 6292805 TI - [Clinico-laboratory diagnosis of chronic ophthalmo-adenovirus infection]. PMID- 6292806 TI - [Ineffectiveness of high-dosage methotrexate in patients with small cell bronchial cancer]. AB - 5 patients with small-cell lung cancer were treated with high-dose methotrexate (1.75-16 g). No significant tumor reduction could be demonstrated. High-dose methotrexate therapy cannot be recommended for the treatment of small-cell lung cancer. Other established and successful regimens are available. PMID- 6292807 TI - Presence of ketohexokinase (EC 2.7.1.3) in human lenses. PMID- 6292808 TI - Chondroblastoma of the temporomandibular joint arising from articular cartilage: a previously unreported presentation of an uncommon neoplasm. PMID- 6292809 TI - Skeletal bone scanning in "Middle East" jaw lymphoma. PMID- 6292810 TI - Human papilloma virus (HPV) antigens in lesions of laryngeal squamous cell carcinomas. AB - Paraffin sections from 36 laryngeal squamous cell carcinomas with histological features of coexistent condylomatous changes (the flat, inverted and papillomatous lesions caused by human papilloma virus; HPV) were stained for HPV antigens with an indirect immunoperoxidase technique. The HPV antiserum used was prepared in guinea pigs immunized against the purified HPV virions from human common wart tissue. Of the tumors studied, 36% showed the presence of HPV antigens. In that, one-third of the flat and inverted lesions and 42% of the papillomatous lesions contained cells with intranuclear HPV-staining reaction. The role of HPV as the etiologic agent of laryngeal squamous cell papillomas is discussed, as is the potential malignant transformation of them. Based on the present results, the previously expressed view on the probable role of HPV in the development of laryngeal squamous cell carcinoma seems to gain further substantiation. PMID- 6292811 TI - Malignant fibrous histiocytoma of the nasal septum. A report of an unusual lesion. AB - A case of malignant fibrous histiocytoma of the nasal septum is described in a 51 year-old woman who has presented with nasal pain, nasal obstruction and a swelling at the base of the nose. Histological examination revealed a malignant fibrous histiocytoma. Total resection of the tumor was performed and, 26 months after operation, the patient was well and free of recurrence or metastasis. This is the first reported case of malignant fibrous histiocytoma of the nasal septum. PMID- 6292812 TI - [Metastasizing cystosarcoma phyllodes]. PMID- 6292813 TI - [Comparative studies of brain scintigraphy using 99mTc-pertechnetate and 99mTc diphosphonate]. PMID- 6292814 TI - Ontogeny of growth factor receptors in the human placenta. AB - The present study was undertaken to further elucidate the role during gestation of insulin, epidermal growth factor-urogastrone (EGF-URO), and a slightly acidic insulin-like growth factor we have referred to as insulin-like activity (ILAs). We examined the ontogeny of the placental receptors for these peptides utilizing membrane fractions (600 x g, 15,000 x g, 100,000 x g) and cytosol (200,000 x g supernatant) prepared by differential centrifugation of early gestation (11-19 wk) and term gestation (38-42 week) human placentae. Aliquots of each membrane fraction were also assayed for 5' nucleotidase activity. The ontogeny of the insulin and EGF receptors closely resembled the pattern of 5' nucleotidase activity, with greater levels seen in term tissue for all membrane fractions assayed. Highest levels of specific binding per mg protein and enzyme specific activity were seen in the 100,000 x g fraction within either gestational age group. The pattern of [125I]-ILAs specific binding was quite different: binding per mg protein was higher in membranes from early gestation placentae regardless of the fraction, with the biggest differential occurring in the 600 X g pellet. As observed with [125I]-insulin and [125I]-EGF, the 100,000 x g pellets from either age group were also enriched in [125I]-ILAs binding. Although neither early gestation nor term cytosol bound [125I]-insulin or [125I]-EGF, binding of [125I]-ILAs to both gestational age groups was clearly demonstrable with significantly higher levels observed in cytosol from early gestation placentae. A knowledge of the mechanisms involved in placental growth may well be a prerequisite to a full understanding of fetal growth control. The demonstration of specific human placental receptors for insulin, EGF-URO, and ILAs as early as 11-19 wk of gestation suggests that these peptides should continue to receive attention as possible modulators of placental growth and function. Furthermore, the high levels of [125I]-ILAs specific binding to membrane and cytosols from early gestation placentae may indicate a more important role for the insulin-like growth factors compared to insulin or EGF-URO during early placental development. PMID- 6292815 TI - Heterogeneous pathways of oxidizing radical production in human neutrophils and the HL-60 cell line. AB - Oxygen-derived free radicals with hydroxyl radical (OH.)-like reactivity are products of the human neutrophil respiratory burst. Such radicals, although dependent on O2-generation arise from complex oxidation reactions that may be independent of an iron-or lactoferrin-catalyzed Haber-Weiss mechanism. Differentiated HL-60 promyelocytic leukemia cells completely deficient in lactoferrin generate oxidizing radicals at a rate greater than that of human neutrophils, indicating lactoferrin-independent pathways for OH. generation. The further heterogeneity of pathways generating OH. activity in neutrophils is indicated by the cell concentration dependence of the reaction, the variability of H2O2 as a precursor reactant, and the various proportions of oxidizing radical activity to O2-detected in human neutrophils stimulated to a variety of agonists. The ethylene assay for oxidizing radical activity may detect different classes of oxidizing species capable of reacting with the aldehyde substrate. The complexity of defining the oxygen-derived radicals of the ethylene assay suggests methodologic difficulties of either quantitating or precisely defining the radicals generated in the respiratory burst by this method. PMID- 6292816 TI - [The Silver-Russell syndrome (clinical, hormonal and cytogenetic study)]. PMID- 6292817 TI - [Case of long-term remission of a Wilms' tumor with metastases to the lungs after the transplantation of an isolated thymus]. PMID- 6292818 TI - Pharmacodynamic effects and possible therapeutic uses of THIP, a specific GABA agonist. AB - THIP (4,5,6,7-tetrahydroisoxazolo[5,4-c]pyridin-3-ol) is a potent and specific GABA receptor agonist which does not influence the GABA uptake system or GABA metabolizing enzymes. The specificity for the GABA receptor is also demonstrated by lack of action on monoaminergic, cholinergic, histaminergic or opiate receptors. Since in recent years GABA receptor stimulants-among others THIP--have become available many have speculated as to what clinical indication GABA-ergic stimulation might be an important element. The first suggestion was that GABA ergic drugs by an inhibitory effect on the dopamine neurons would improve the antischizophrenic effect of neuroleptics and improve tardive dyskinesia. Furthermore, studies on brains of deceased Parkinson and Huntington's chorea patients have demonstrated a low level of GABA and its synthesizing enzyme glutamic acid decarboxylase (GAD) in the basal ganglia. Also in epilepsy and diseases with dementia a deficit in the GABA system has been proposed. Therefore a therapeutic strategy for these diseases may be supplementary treatment with drugs which increase GABA receptor activity. Furthermore, recent results in humans have shown that GABA agonists perhaps also could be of benefit in mania and depressions. When considering the neurophysiological elements of nociception and muscle tone it is also reasonable to suggest that GABA-ergic stimulation may reduce pain perception and muscle tone. PMID- 6292819 TI - Angiotensin converting enzyme inhibitors. AB - Inhibition of angiotensin converting enzyme (ACE) in patients suffering from renovascular hypertension results in lowering of the blood-pressure. The development of captopril, an orally active ACE inhibitor and the structure activity relationship of captopril analogues are described. Some side effects of captopril were attributed to the presence of an SH-group in the molecule. This led to the development of a series of non-mercapto ACE inhibitors derived from captopril. One of them, enalapril, a potent ACE inhibitor with a long duration of antihypertensive action, is now tested in clinical trials. PMID- 6292820 TI - Postsynaptic currents in different types of frog muscle fibre. PMID- 6292821 TI - The effects of intracellular protons on the electrical activity of single ventricular cells. AB - Acidic or alkaline solutions were injected into single guinea-pig ventricular cells and the effects of cytoplasmic proton concentrations ([H]i) on their electrical activity were examined. Injections of low pH (3.7-4.7) solutions shortened the action potential and depressed the plateau by decreasing isi and increasing the outward current at the plateau potentials. Injections of high pH (9.3-9.7) solutions had the opposite effects on the action potential and the membrane currents. A decremental oscillatory current, similar to the strophanthidin-induced transient inward current, was generated upon injecting large amount of low pH solutions. Injections of high pH solution abolished this current. Alterations in [Ca]i also mimiced the changes of the action potential caused by H+-injections. Comparison of the effects of high pH and EGTA injections, however, disclosed marked different actions of these agents on the inactivation process of isi. Therefore, it may be concluded that a combination of a direct effect of the alterations in [H]i and an effect due to changes in [Ca]i secondary to proton injections, causes the above electrical changes of single ventricular cells. PMID- 6292822 TI - Poorly selective cation channels in the skin of the larval frog (stage less than or equal to XIX). AB - The abdominal skin of bullfrog larvae (Rana catesbeiana) was placed in an Ussing type chamber, and its transepithelial electrical parameters were recorded with mucosal solutions of different ionic composition. With "K+-like" cations (K+, NH+4, RB+, Cs+) the power spectra of the fluctuations in short-circuit current displayed a Lorentzian component (fc = 30 - 40 Hz). The relaxation noise could be suppressed by addition of the K+ -channel blockers Ba2+ and TEA to the mucosal solution. Also, in presence of the ionophore antibiotic nystatin the Lorentizian noise was abolished. The Na+ -channel probes amiloride and benzimidazolyl-2 guanidine (BIG) both enhanced the relaxation noise obtained with the K+-like cations but, with Na+ and Li+, also caused the rise of a relaxation component above the background noise. In presence of amiloride or BIG, the addition of Ba2+, TEA and nystatin still abolished the Lorentizian noise. It can be concluded that the relaxation-noise source is located in the apical cell membranes of the tadpole skin. These spontaneously fluctuating cation channels do not seem to strictly discriminate between K+-like ions (K+, NH+4, Rb+, Cs+) and Na+-like ions (Na+, Li+). On the other hand, well-known specific probes for K+ channels (Ba2+, TEA) and for Na+ channels (amiloride, BIG) interact with this apical cation channel. It is possible that the poorly selective channel plays a role in the ontogenesis of the specific Na+ transport in the maturing frog skin. PMID- 6292824 TI - Electrical and mechanical properties and neuro-effector transmission in the smooth muscle layer of the guinea-pig ileocecal junction. AB - Electrical and mechanical properties and neuroeffector transmission were studied in circular strips of smooth muscle taken from the ileocecal junction of guinea pigs in relation to sphincter action, using the microelectrode, and tension recording methods. The membrane potential of the smooth muscle was low (-43mV) compared with the membrane potential of circular muscle cells of the ileum or caecum (-58 mV or 62 mV). Only small populations of the muscle cells (about 5%) generated spontaneous action potentials. Field stimulation of the tissue produced an initial slight relaxation followed by a contraction, and the mechanical responses were accompanied by membrane hyperpolarization (i.j.p.) followed by repolarization with rebound spikes. Treatment with atropine increased the amplitude of i.j.p.s and decreased the amplitude of the rebound repolarization. Propranolol or phentolamine did not affect the amplitude of i.j.p., however, phentolamine slightly reduced the amplitude of the rebound repolarization. These results indicate that the ileocecal junction is predominantly controlled by non adrenergic, non-cholinergic inhibitory nerve fibres and that the distribution of adrenergic and cholinergic excitatory nerve fibres is sparse. PMID- 6292825 TI - [CT arteriography in diagnosis of insulinomas]. PMID- 6292823 TI - Tubular transport processes in proximal tubules of hypothyroid rats. Lack of relationship between thyroidal dependent rise of isotonic fluid reabsorption and Na+ -K+ -ATPase activity. PMID- 6292826 TI - [Value of the artificial pancreas in the diagnosis of organic hypoglycemia]. PMID- 6292827 TI - Pathophysiological mechanisms operating in the development of myelofibrosis: role of megakaryocytes. AB - In this article current concepts on the regulation of bone marrow collagen are reviewed and a hypothesis regarding the mechanisms leading to marrow fibrosis associated with Primary Myelofibrosis (PMF) is presented. Type I and type III collagen, products of marrow fibroblasts, are the main constituents of myelofibrotic tissue and megakaryocytes are the predominant cells proliferating in PMF. There is evidence for the clonal nature of the hematopoietic cell proliferation and the secondary origin of myelofibrosis. Also, evidence exists indicating that defective megakaryocyte maturation, i.e. ineffective megakaryocytopoiesis occurs in patients with PMF. It is postulated that ineffective megakaryocytopoiesis leads to an excessive concentration of megakaryocyte components in the marrow intercellular space and that the development of marrow fibrosis involves mainly 2 megakaryocytic products: growth factor and factor 4. The growth factor stimulates fibroblast proliferation and collagen secretion. Factor 4 inhibits the activity of the enzyme collagenase. Thus, the imbalance between increased collagen production and decreased collagen degradation leads to an excessive deposition of collagens in bone marrow matrix. PMID- 6292828 TI - Erythrocyte membrane phosphorylation in sickle cell disease. AB - Phosphorylation of erythrocyte membrane proteins was determined in patients with homozygous sickle cell disease. After incubation of ghosts with gamma-32P ATP, proteins were submitted to SDS-polyacrylamide gel electrophoresis. Three salient features appeared: (i) a decreased phosphorylation of spectrin bands; (ii) a significantly increased phosphorylation (P less than 0.001) of bands 4(5) and 4(8) in the absence of cAMP and (iii) a significantly increased phosphorylation (P less than 0.001) of bands 7 and 8, both in the absence and the presence of cAMP. Studies on reticulocyte rich blood showed that the first change appeared to be specifically related to the disease, whilst the second resulted from the rejuvenation of the red cell population. No definite conclusion could be drawn for the third alteration. PMID- 6292829 TI - The infectivity of adenovirus genomes lacking DNA sequences from their left-hand termini. AB - Deletions extending various distances into the left-hand terminal DNA sequences of the adenovirus type 2 (Ad2) genome were generated in a plasmid containing a cloned fragment spanning from 0 to 4.9 map units. The altered Ad2 DNA sequences were introduced into viral genomes by ligating a plasmid-derived fragment, which included the sequences extending to 3.8 map units, to the 3.8-100 map unit fragment generated by XbaI cleavage of the DNA of the Ad5 variant, d1309 (N. Jones and T.Shenk, Cell 17 683-689, 1979). The infectivity of the ligation products was studied by transfection of line 293 cells. Genomes lacking 11, 40, or 51 nucleotides from their left-hand termini, or containing an additional 18dG residues linked to this position were infectious, and analysis of the progeny virus genomes demonstrated that the structure of these modified termini had been restored to normal. In contrast, genomes from which the first 160 base pairs (bp), including the entire 102 bp left hand inverted terminal repeat (ITR), had been removed were non-infectious. The results indicate that the ITRs present at the opposite ends of transfecting DNA molecules are able to interact in vivo, and enable the production of viable viruses containing corrected left-hand terminal sequences. Possible mechanisms for this interaction are discussed. PMID- 6292830 TI - Prostatic steroid binding protein: organisation of C1 and C2 genes. AB - Prostatic steroid binding protein, whose expression is stimulated by androgens, consists of two subunits: one containing the polypeptides C1 and C3 and the other containing C2 and C3. We have characterised genomic clones containing the C1 and C2 genes by restriction enzyme analysis and DNA sequencing. Both genes are 3.2 Kb, have similar exon/intron arrangements and share considerable DNA sequence homologies in their coding regions, intervening sequences and 5' upstream DNA sequence which suggests that they have probably arisen from the duplication of an ancestral gene. The 5' termini of C1 and C2 mRNA have been mapped; the sequence TATAAA appears 30 nucleotides upstream but a CAAT-like sequence at -60 - -80 is absent. Finally, homologous human genes have not been detected. PMID- 6292832 TI - Deletion loop mutagenesis: a novel method for the construction of point mutations using deletion mutants. AB - Deletion loop mutagenesis is a new, general method for site-directed mutagenesis that allows point mutations to the introduced within a sequence of DNA defined by a previously isolated deletion mutant. Wild type and deletion mutant DNA are cloned into a bacterial plasmid and each is cleaved with a different single cut restriction enzyme. Heteroduplexes are formed between the two DNAs to produce circular molecules containing a nick in each strand and a single-stranded deletion loop. The deletion loops are mutagenised using sodium bisulphite and the DNA transfected directly into a uracil repair deficient strain of Escherichia coli. Up to half of the resultant clones contain DNA produced by replication of the wild-type length strand and bear mutations exclusively within the target area. An example is given in which a deletion mutant lacking 21 nucleotides from the region coding for SV40 large-T was used. Eight of the possible nine target cytosine residues were mutagenised. The method described is specific, efficient and simple. PMID- 6292833 TI - Structure-function relationship of Rous sarcoma virus leader RNA. AB - Cells infected by RSV synthesize viral 35S RNA as well as subgenomic 28S and 22S RNAs coding for the Env and Src genes respectively. In addition, at least the 5' 101 nucleotides of the leader are also conserved and we have shown previously that this sequence contains a strong ribosome binding site (J.-L. Darlix et al., J. Virol. 29, 597). We now report the RNA sequence of Rous Sarcoma virus (RSV) leader RNA and propose a folding of this 5' untranslated region which brings the Cap, the initiation codon for Gag and the strong ribosome binding site close to each other. We also show that ribosomes protect a sequence just upstream from initiator Aug of Gag in vitro, and believed to interact with part of the strong ribosome binding site according to the folding proposed for the leader RNA. PMID- 6292831 TI - Epsilon globin gene transcripts originating upstream of the mRNA cap site in K562 cells and normal human embryos. AB - RNA transcribed from the human epsilon globin gene was studied in K562 cells and human embryos of 5-10 weeks gestation. Using both primer extension and S1 analysis RNA molecules were found which extend 53 +/- 1 bp, 55 +/- 1 bp and 270 +/- 1 bp upstream from the first coding base and are colinear with the gene. The first pair of molecules represent transcripts initiating at the canonical cap site but the other species represents a transcript over 200 nucleotides longer which initiates upstream of the CAAT and ATAA boxes. PMID- 6292834 TI - Yeast RNA polymerase II transcription of circular DNA at different degrees of supercoiling. AB - Purified yeast RNA polymerase II was tested for transcriptional activity as a function of the degree of circular DNA supercoiling. Chimaeric plasmids P30 and P31 both containing inserts from the yeast transposable element TY1 cloned in pBR322 and the vector pBR322 were used as templates. For pBR322 the transcriptional activity increases about 4 fold from the fully relaxed covalently closed circles to the native supercoiled forms, further supercoiling having no effect on transcription. P30 shows a 5 fold increase of transcriptional activity reaching a plateau at the native supercoiled conformation. However, at an intermediate degree of supercoiling (sigma = 0.024), transcription decreases to a value close to zero. P31 too exhibits a conformation (sigma = 0.014) in which there is a drop of transcriptional activity. Furthermore, a 10 fold increase of transcription is obtained at the higher values of superhelix density. Both kinetic and autoradiographic experiments confirm the existence of DNA conformations that can inhibit "in vitro" transcription. PMID- 6292835 TI - Molecular structure of uvrC gene of Escherichia coli: identification of DNA sequences required for transcription of the uvrC gene. AB - We have carried out experiments to identify the regulatory regions of the uvrC gene of Escherichia coli. A uvrC+ plasmid, pUV7, containing the intact transcriptional unit for the uvrC gene, was used to subclone either the structural gene or combinations of the structural gene and 5'-flanking sequences. The plasmids so constructed were tested for ability to restore UV-resistant phenotype to uvrC- cells as an indication of expression of the uvrC gene. The chromosomal DNA in plasmid pUV7 was probed for strong binding with E. coli RNA polymerase in an attempt to identify a restriction fragment which bears the regulatory sequences for the uvrC transcriptional unit. The results indicate that DNA sequences at least 0.9 Kb upstream from the structural gene, but not the 5' proximal sequences, regulate expression of the uvrC gene. Analysis of protein synthesis encoded by plasmid pUV7 and its derivatives suggest that there may be another gene that lies between the promoter and the uvrC gene and codes for a 27,000-Mr protein. The relation of this gene to uvrC function is not clear. PMID- 6292836 TI - Sequence and secondary structure of mouse 28S rRNA 5'terminal domain. Organisation of the 5.8S-28S rRNA complex. AB - We present the sequence of the 5' terminal 585 nucleotides of mouse 28S rRNA as inferred from the DNA sequence of a cloned gene fragment. The comparison of mouse 28S rRNA sequence with its yeast homolog, the only known complete sequence of eukaryotic nucleus-encoded large rRNA (see ref. 1, 2) reveals the strong conservation of two large stretches which are interspersed with completely divergent sequences. These two blocks of homology span the two segments which have been recently proposed to participate directly in the 5.8S-large rRNA complex in yeast (see ref. 1) through base-pairing with both termini of 5.8S rRNA. The validity of the proposed structural model for 5.8S-28S rRNA complex in eukaryotes is strongly supported by comparative analysis of mouse and yeast sequences: despite a number of mutations in 28S and 5.8S rRNA sequences in interacting regions, the secondary structure that can be proposed for mouse complex is perfectly identical with yeast's, with all the 41 base-pairings between the two molecules maintained through 11 pairs of compensatory base changes. The other regions of the mouse 28S rRNA 5'terminal domain, which have extensively diverged in primary sequence, can nevertheless be folded in a secondary structure pattern highly reminiscent of their yeast' homolog. A minor revision is proposed for mouse 5.8S rRNA sequence. PMID- 6292837 TI - Synthesis and biological properties of 2'-deoxy-5-vinyluridine and 2'deoxy-5 vinylcytidine. AB - Rapid and efficient syntheses for the preparation of 2'-deoxy-5-vinyluridine and 2'-deoxy-5-vinylcytidine are described starting from nucleoside precursors. Contrary to some previous reports, 2-deoxy-5-vinyluridine has be found to be quite stable under normal laboratory conditions but when tested in animals shows neither toxicity nor anti-leukemic (L1210 cells) or anti-parasitic (Plasmodium berghei) activity. 2'-Deoxy-5-vinylcytidine appears to polymerise readily. It is much less toxic to cell cultures than 2'-deoxy-5'vinyluridine but is almost as active against herpes virus replication (ID50:0.2 microgram/ml) for both type 1 and type 2 herpes virus (selectivity index:225). PMID- 6292838 TI - Long terminal repeat (LTR)-derived recombination of retroviral DNA: sequence analyses of an aberrant clone of baboon endogenous virus DNA which carries an inversion from the LTR to the gag region. AB - Nucleotide sequences of a cloned proviral DNA of baboon endogenous virus M7 were analyzed, which carried an internal inversion. The inversion of 2.2 kilobase pairs was occurred between the junction of two tandem LTRs and a site locating in the p30 region of the gag gene. The ATAA sequence was a target for recombination generating the inversion, which was duplicated at both ends of the inverted segment. AAA and CA were lost at the 5'- and 3'-ends of the LTRs by the inversion, respectively. On both sides of the target sequence, long AG-rich stretches were detected, which may specify the site of recombination together with the target sequence. The characteristic base changes in the inversion are concluded to result from an illegitimate recombination associated with LTRs, as well as in case of provirus integration into the host cell DNA. We propose and discuss models to explain the processes of recombination to generate both inversion and integration. PMID- 6292839 TI - Cloning and sequence analysis of a cDNA plasmid for one of the rat liver glutathione S-transferase subunits. AB - We describe the construction and characterization of a cDNA plasmid for one of the rat liver glutathione S-transferase subunits. Poly(A)-RNA isolated from rat livers was enriched for glutathione S-transferase mRNA activity and used as templates to synthesize double stranded cDNA. The double stranded cDNAs were annealed to pBR322 through terminal deoxynucleotidyl transferase generated GC tails followed by transformation into E. coli. Several candidate clones were selected by colony hybridization using polynucleotide kinase labeled liver and testis poly(A)-RNA probes. These candidate clones were further characterized by hybrid-selected translation of mRNA followed by immunoprecipitation and SDS gel electrophoresis. The positive clone, pGTR112 was mapped with restriction endonuclease analysis and sequenced by the chemical method of Maxam and Gilbert. The largest upen reading frame contains 142 amino acids very rich in Arg and Lys residues. The C-terminal residue phenylalanine of this open reading frame is consistent with what was reported for one of the ligandin subunits by Bhargava et al., (J. Biol. Chem. 253, 4116-4119, 1978). Among the 352 nucleotides covered by both pGTR112 and pGST94 described by Kalinyak and Taylor (J. Biol. Chem. 257, 523 530, 1982), there are only 9 nucleotide differences resulting in four changes of amino acid sequences. PMID- 6292841 TI - Accurate in vitro splicing of human beta-globin RNA. AB - Human beta-globin RNA transcribed from an exogenous DNA template is spliced in vitro by concentrated whole cell extracts from HeLa cells. Using the primer extension technique, we have shown that the small intervening sequence is spliced accurately and that the sequence of the product across the splice junction is identical to that of beta-globin mRNA prepared from human reticulocytes. The efficiency of the splicing reaction is low. The RNA transcript containing both introns and terminated upstream from the polyadenylation site is spliced most efficiently. The transcript which is terminated downstream from the polyadenylation site is not spliced at all. Thalassemic beta-globin RNA which carries an extra splice site in the small intron is also spliced, albeit with a low yield. PMID- 6292842 TI - Assembly of semihistone A24. AB - Nucleosomal semihistone A24 (uH2A) is composed of histone H2A and ubiquitin peptides. The kinetics of incorporation of nascent H2A and ubiquitin into A24 of transformed chicken lymphocytes (MSB cells) have been examined by peptide mapping, COOH-terminus analysis, density labeling and isopycnic centrifugation of chromosomal proteins. We find that newly synthesized H2A is rapidly conjugated to ubiquitin. SDS-PAGE analysis of fractionated density gradients suggests, however, that newly synthesized ubiquitin becomes bound nonselectively to both new and preexisting H2A molecules. PMID- 6292840 TI - mRNA-deficient beta o-thalassemia results from a single nucleotide deletion. AB - The beta-globin gene of a patient with mRNA-deficient beta o-thalassemia has been sequenced. We find a single nucleotide deletion in amino acid codon 44 that produces a UGA terminator at codon 60. We have previously shown that the beta globin mRNA of this patient is correctly spliced and polyadenylated, but rapidly turns over with a half-life of less than 30 min. We suggest that the rapid mRNA turnover is influenced by the deletion of this single nucleotide as well as by the nonsense codon. PMID- 6292843 TI - Structure of 1.71 lb gm/cm(3) bovine satellite DNA: evolutionary relationship to satellite I. AB - The Eco RI fragments from the 2600 bp repeating unit of 1.711b gm/Cm(3) bovine satellite DNA were cloned in pBR322. The structure of the repeat unit was determined and compared to bovine satellite I DNA (rho CsCl = 1.715 gm/cm(3)). All of the DNA in the 1402 bp repeat of satellite I is represented in the sequence of the 2600 bp 1.711b gm/cm(3) repeat. The difference between the two repeats is due to a 1200 bp piece of DNA (INS) residing in the middle of the 1.711b gm/cm(3) repeat. The INS is AT-rich and has some repetitive components; it bears only limited similarity to the structure of eukaryotic transposable elements. We propose that the 1.711b gm/cm(3) satellite DNA arose via the amplification of a 1.715 gm/cm(3) satellite repeat altered by a 1200 bp insertion of DNA. PMID- 6292844 TI - DNA polymerase alpha from the nuclear matrix of cells infected with simian virus 40. AB - The nuclear matrix prepared from normal, simian virus 40 (SV40)-infected, and SV40-transformed cells contained DNA polymerase activities. Approximately 12% of the total DNA polymerase activities in isolated nuclei remained with the nuclear matrix. alpha-polymerase was the major matrix DNA polymerase activity as judged by sensitivity to various inhibitors: aphidicolin, dideoxy-TTP, and N ethylmaleimide. Approximately 2-4 fold higher DNA polymerase activity was detected in matrices obtained from lytically infected and virus-transformed cells than that found in normal cells. In lytically infected cells, 30-50% of the matrix-bound DNA polymerase activity solubilized by sonication co-sedimented with majority of the matrix T-antigen, and was co-precipitated with anti-T sera. The results suggest that alpha-polymerase and viral T-antigen may form a functional complex in the matrix. PMID- 6292846 TI - Sequence homologies of diverse length tandem repetitions near ends of vaccinia virus genome suggest unequal crossing over. AB - The 180,000 base pair (bp), covalently closed, linear duplex DNA genome of vaccinia virus contains a 10,000 bp inverted terminal repetition within which are one set of 13 and one set of 18 tandem 70 bp repeating units. A 967 bp segment containing the innermost 70 bp repeat and an adjacent region notable for a scarcity of restriction endonuclease sites has been sequenced. This was facilitated by the cloning of TaqI and partial TaqI fragments in pBR322. We found that the innermost 70 bp repeat overlaps one of two adjacent 125 bp repeats, following which are eight repeats of 54 bp, parts of 54 bp and 70 bp repeats, and four consecutive 6 to 7 bp repeats. The 70, 125, and 54 bp repeating units have extensive sequence homologies and redundancies that suggest evolution by unequal crossing over. Schemes whereby unequal crossovers of 54 bp repeats lead to a recombinant segment 86% homologous to the 125 bp repeat and unequal crossovers of 125 bp repeats lead to a recombinant segment 94% homologous to the 70 bp repeat were considered. This propensity for sequence divergence should provide a useful marker for comparing the relatedness of poxviruses. PMID- 6292845 TI - Conversion of simian virus 40 DNA to ordered nucleoprotein structures by extracts that direct accurate initiation by eukaryotic RNA polymerase II. AB - Interaction of SV40 DNA with three different HeLa cell extracts capable of directing correct initiation of transcription leads to the formation of ordered nucleoprotein complexes that are structurally similar to SV40 minichromosomes and eukaryotic chromatin. These nucleoprotein complexes can be conveniently purified by band sedimentation or gel filtration. Their sedimentation and elution properties resemble those of SV40 minichromosomes. Electron microscopy of purified complexes shows beaded structures that are sensitive to proteases, resulting in recovery of naked, largely undegraded DNA. Contour lengths and compaction ratios of these nucleoprotein complexes are similar to those of authentic SV40 minichromosomes. Their digestion patterns with micrococcal nuclease and pancreatic DNase I resemble those of SV40 minichromosomes. Such nucleosome-like structures can also be obtained with linear SV40 DNA. Unlike nucleosomes, no histones could be detected in the purified nucleoprotein complexes. Non-histone chromosomal protein fractions (high mol. wt. and free of high mobility group proteins) prepared from the HeLa cell extracts can also generate similar ordered structures. We conclude that ordered nucleoprotein structures with certain common characteristics can be formed by interaction of DNA with non-histone chromosomal proteins as well as with histones. Only the former structures are generated in currently used cell-free transcription systems. It appears that only those purified nucleoprotein complexes containing the promoter can be actively transcribed in the presence of additional cell-free extract, suggesting that such structures and their protein components may be important in transcription. PMID- 6292847 TI - Structure of cloned delta-globin genes from a normal subject and a patient with delta-thalassemia; sequence polymorphisms found in the delta-globin gene region of Japanese individuals. AB - The delta-globin genes of a normal Japanese and a Japanese patient with homozygous delta-thalassemia were cloned, and the nucleotide sequence of a region including the gene was determined. Comparison of the nucleotide sequences of these two individuals with that of pH delta 1, delta-globin clone from the gene library constructed by Maniatis et al., showed differences in the large intervening sequence (IVS 2), at positions 137, 151, 186, 188, 291, 292 and 540 as one base substitutions, at 339 and 823 as one base additions, at 548 as a one base deletion, and a 9 bp duplication between positions 651 and 659, and differences in the 3'-flanking sequence at 51 and 98 nucleotides 3' to the AATAAA sequence. However, in the region studied, no differences was observed in the nucleotide sequences of the normal subject and the patient with delta thalassemia. Therefore, these differences may represent polymorphisms of the delta-globin gene present in Japanese individuals. These data suggest that IVS 2 is more divergent than other regions, and that a DNA region(s) other than the globin gene may affect expression of the gene. PMID- 6292849 TI - New restriction endonucleases from Acetobacter aceti and Bacillus aneurinolyticus. AB - Two restriction endonucleases with new sequence specificities have been isolated from Acetobacter aceti IFO 3281 and Bacillus aneurinolyticus IAM 1077 and named AatII and BanII, respectively. Based on analysis of the sequences around the restriction sites, the recognition sequences and cleavage sites of these endonucleases were deduced as below: (formula; see text) PMID- 6292848 TI - Molecular cloning of retrovirus-like genes present in multiple copies in the Syrian hamster genome. AB - Endogenous retrovirus-like sequences homologous to intracisternal type-A particle (IAP) genes, which are present in the inbred mouse (Mus musculus) genome, were cloned from a Syrian hamster gene library. A typical hamster IAP gene was 7 kb long and segments homologous to long terminal repeat (IAP) sequences present in Mus musculus IAP genes were located at both ends of the gene. Contrary to the pattern found in the Mus musculus IAP genes, the organization of the cloned hamster IAP genes was not markedly polymorphic and deletion was not observed among these cloned genes. A sequence about 0.8 kb long and located close to the 3' end of the hamster IAP gene was well conserved in both IAP gene families, although they showed less overall homology with one another. The reiteration frequency of the hamster IAP genes was calculated to be 950 copies per haploid genome. Since such IAP genes with the above properties were not found in the genome of the Chinese hamster, whose progenitors diverged from those of the Syrian hamster about 7.5 Myr ago, the integration of a huge number of Syrian hamster IAP genes must have occurred subsequent to such divergence. PMID- 6292850 TI - pUR 250 allows rapid chemical sequencing of both DNA strands of its inserts. AB - A multipurpose plasmid, pUR 250, has been constructed. It contains seven unique cloning sites (Hind III, Xba I, Sal I, Acc I, Hinc II, BamH I and EcoR I) in a small region of its lac Z gene part. Insertion of foreign DNA into the plasmid can be easily detected. Plasmid DNA isolated from a clone by a rapid method can be used to determine the DNA sequence of the insert from both ends directly without isolation of labeled fragment. PMID- 6292851 TI - High level synthesis in Escherichia coli of the Bacillus subtilis phage phi 29 proteins p3 and p4 under the control of phage lambda PL promoter. AB - The Hind III G fragment from the Bacillus subtilis phage phi 29 DNA, inserted downstream from the bacteriophage lambda promoter PL carried by a pBR322 derivative plasmid (pPLc28), directed the synthesis in E. coli of two proteins of apparent molecular weight 27500 and 12500. With the use of the recombinants obtained with the DNA from mutants sus3(91) and sus4(56), the two proteins were identified as a modified p3 (p3'), the protein covalently linked to the 5' ends of phi 29 DNA, and p4, responsible for the phi 29 late transcription, respectively. Under the best conditions used, proteins p4 and p3' were produced in E. coli from the cloned DNA fragments in an amount corresponding to approximately 30% and 6% of total de novo protein synthesis, respectively. PMID- 6292852 TI - Nucleotide sequence of the early genes 3 and 4 of bacteriophage phi 29. AB - The nucleotide sequence of an early region of the phi 29 genome has been determined. The sequenced region includes genes 3 and 4, which code for the protein covalently linked to the 5' ends of phi 29 DNA and the protein involved in the control of late transcription, respectively. The position and nature of the mutations of mutants sus3(91) and sus4(56) has also been determined. PMID- 6292853 TI - Genes for tRNALys5 from Drosophila melanogaster. AB - The sequences of two cloned genes from Drosophila which hybridize with tRNALys5 are reported. One gene, in plasmid pDt39, has a sequence which corresponds to the sequence of tRNA. The other gene, in pDt59R, differs in three nucleotides pairs. Both plasmids are transcribed in vitro with extracts of Drosophila Kc cells to give full-sized tRNA precursors with four additional nucleotides at the 5'-end as well as truncated molecules containing 35 nucleotides. This premature termination occurs in a block of four T residues within the mature coding region. Sequences flanking the tRNA genes show little in common except for the blocks of five or more T-residues beyond the 3'-end of the gene. pDt39 hybridizes to 84AB on the polytene chromosomes of Drosophila and pDt59R hybridizes to 29A. PMID- 6292854 TI - Analysis of chromatin structure and DNA sequence organization: use of the 1,10 phenanthroline-cuprous complex. AB - Limited treatment of Drosophila nuclei with the 1,10-phenanthroline-cuprous complex leads to rapid production of nucleosomal ladders indistinguishable from those obtained by micrococcal nuclease digestion. An investigation of the preferential sites of cleavage of protein-free DNA at locus 67B1 surprisingly indicated that both reagents recognized very similar features. Thus, a virtually identical pattern of preferential cleavages was generated over a 12 kb fragment encoding four transcripts at this locus. The distribution of cleavage sites was highly non-random, with major sites falling in the spacers between the genes. Both reagents cleaved certain chromatin-specific sites near the 5' ends of the genes. However, an analysis of preferential cleavages at the sequence level did not reveal the same close correspondence. We suggest that both reagents can recognize some localized secondary structural features of the DNA and that the particular distribution of sequences present at this locus results in a distinctive pattern of cleavage sites that delineates gene and spacer segments. PMID- 6292855 TI - Multiple arrangements of the human embryonic zeta globin genes. AB - Rearrangements which are most readily explained by homologous crossover between misaligned segments of DNA in the region of the human embryonic zeta (zeta) globin genes have been identified in individuals of three different racial origins. These recombination events have resulted in a surprisingly high prevalence of chromosomes with single (0.4%) and triplicated (1.3%) zeta genes with apparently no significant effect on the phenotype. PMID- 6292856 TI - The structure of the gene coding for the phosphorylated ribosomal protein S10 in yeast. AB - From previous studies on cloned yeast ribosomal protein genes we obtained evidence that a large number of them contain an intron [Bollen et al. (1982) Gene 18, 29-38]. In the temperature-sensitive rna2-mutant transcription of these genes leads to the accumulation of precursor RNAs at the restrictive temperature. These precursor mRNAs are several hundreds of nucleotides longer than the respective mature mRNAs. The split character of one of these ribosomal protein genes, viz. the gene coding for the major phosphorylated small-subunit protein S10, was further established by sequence analysis. The intervening sequence interrupts the coding sequence after the second codon and has a length of 352 nucleotides. Genomic Southern hybridizations with a DNA fragment carrying part of the S10-gene revealed that this gene is duplicated on the yeast genome. The molecular weight of S10 as deduced from the sequence analysis was estimated to be 31462 dal. Comparison of the N-terminal aminoacid sequence of the yeast ribosomal protein S10 with that of ribosomal protein S6 from rat liver revealed a striking homology between both proteins. Moreover, at the C-terminal end of the yeast ribosomal protein the sequence Arg-Ala-Ser-Ser-Leu-Lys is present which is very similar to the phosphorylation site of the rat liver protein S6. PMID- 6292858 TI - Yeast RNA polymerase I binds preferentially to A+T-rich linkers in rDNA. AB - Restriction fragments of yeast rDNA retained by purified RNA polymerases on nitrocellulose filters were analysed by gel electrophoresis. The EcoRI fragment B was preferentially retained by RNA polymerase I, but not by RNA polymerase III. The in vivo initiation sites for both polymerases are located within this fragment. Further analysis indicated that the preferred binding site for RNA polymerase I is highly AT-rich regions rather than a true promoter. The reported selective in vitro transcription of rDNA by purified yeast RNA polymerase I could then be explained by this preferential binding. PMID- 6292859 TI - An analysis of cosmid clones of nuclear DNA from Trypanosoma brucei shows that the genes for variant surface glycoproteins are clustered in the genome. AB - Trypanosoma brucei contains more than a hundred genes coding for the different variant surface glycoproteins (VSGs). Activation of some of these genes involves the duplication of the gene (the basic copy or BC) and transposition of the duplicate to an expression site (yielding the expression-linked copy or ELC). We have cloned large fragments of genomic DNA in cosmid vectors in Escherichia coli. Cosmids containing the BCs of genes 117, 118 and 121 were readily obtained, but DNA containing the ELCs was strongly selected against in the cosmid and plasmid cloning systems used. We have analysed the distribution of VSG genes in the genome using probes for the sequences at the edges of the transposed segment which are partially homologous among these genes. In genomic cosmid clone banks, about 9% of all colonies hybridize with probes from the 5'- and 3'-edges of the transposed segment, showing that these sequences are linked in the genome. Moreover, the 117 and 118 BC cosmids contain several additional putative VSG genes in tandem, as deduced from hybridization and sequence analyses. We conclude that the VSG genes are highly clustered and share common sequences at the borders of the transposed segment. PMID- 6292857 TI - Loss of type I procollagen gene expression in SV40-transformed human fibroblasts is accompanied by hypermethylation of these genes. AB - Transformation of human lung fibroblasts (WI-38) by Simian Virus 40 (SV40) resulted in a decline of 25-30% in the amount of secreted collagen. The collagen produced by the transformed fibroblasts contained no type I collagen (i.e. alpha 1(I) and alpha 2 chains), which was the major collagen component produced by untransformed fibroblasts. Measurement of the procollagen mRNA levels by dot hybridization with nick-translated procollagen-cDNA clones showed that the absence of type I collagen was due to the absence of alpha 1(I) and alpha 2 procollagen mRNAs. This result was confirmed by hybridization of cDNA to total RNA with southern blots of the procollagen clones. To clarify the mechanism by which type I procollagen gene transcription is abolished in transformed cells, the methylation patterns of the alpha 1(I) and alpha 2 procollagen genes in normal and SV40-transformed fibroblasts were compared, using the chicken alpha 1(I) and alpha 2 procollagen-cDNA clones as probes. Methylated sites were detected by means of the restriction endonuclease isoschizomers HpaII and MspI. Methylation of the procollagen alpha 1(I) and alpha 2 genes was increased in the SV40-transformed fibroblasts, concurrently with the loss of type I collagen synthesis. DNA methylation may thus contribute to altered regulation of gene expression upon cell transformation. PMID- 6292860 TI - IS3 can function as a mobile promoter in E. coli. AB - We had shown previously (3) that the E. coli argE gene could be turned-on by an IS3 element inserted in orientation II near the 5' end of the gene. Here we show that this effect is due to the presence of an outward promoter located on IS3. The exact site of insertion of IS3 was determined by DNA sequencing. Using the S1 nuclease mapping technique with in vivo transcribed RNA we located the promoter responsible for argE transcription on IS3 itself outside the region involved in the inverted repeats of this element. IS3 may therefore be considered as a mobile promoter. PMID- 6292861 TI - Purification of ribonuclease H as a factor required for initiation of in vitro Co1E1 DNA replication. AB - Escherichia coli ribonuclease H was purified to near-homogeneity and identified as the only additional factor required for initiation of in vitro Co1E1 DNA replication from the unique origin by RNA polymerase and DNA polymerase I. Both ribonuclease H activity and stimulating activity for Co1E1 DNA synthesis comigrate with the single protein band in gel electrophoresis. These two activities coincide throughout the process of purification. Some DNA synthesis takes place on covalently closed-circular DNA molecules other than Co1E1 DNA with the three purified enzymes. This DNA synthesis is suppressed by an Escherichia coli single-strand DNA binding protein and/or a high concentration of ribonuclease H. Negative superhelicity of template DNA is required for efficient primer formation. No evidence that supports involvement of ribonuclease III in initiation of Co1E1 DNA replication or its regulation was found. PMID- 6292862 TI - Comparative DNA analysis of three South American marsupials. AB - Published information on marsupials DNA is limited to a group of species belonging to only one genus. No previous reports have been written on South American species. In this paper we characterize the DNA of three out of the four marsupials found in Uruguay. Analytical and preparative ultracentrifugations in neutral CsCl gradients, including four intercalating agents and in Cs2SO4 gradients in presence of increasing amounts of Hg++ ion did not allow us to separate any satellite fraction. The buoyant density of the unique peak measured in CsCl gradients was in every case 1.697 g/cc with a G-C content of 37.7%. Digestion of total DNA with 11 restriction endonucleases produced a different pattern of bands for the three species, although some possible homologies could be established. Hybridization with 32P-rRNA of Southern blots of the gels containing digested DNAs demonstrated that the repeated sequences evidenced do not correspond to the ribosomal cistrons. PMID- 6292863 TI - A Drosophila ribosomal protein gene is located near repeated sequences including rDNA sequences. AB - We have isolated a cloned segment of Drosophila genomic DNA containing a ribosomal protein gene. Hybridization analysis of the DNA in this clone indicates a complex organization of repeated elements within this cloned segment. At least one of these repeated elements is homologous to regions of rDNA. Restriction analysis of the clone shows that some of the repeated elements are present as tandem duplications and in scattered locations within the cloned DNA segment. There are also three non-ribosomal protein genes contained in this clone, each of which is expressed along with the ribosomal protein gene into RNA species present in Drosophila embryos. PMID- 6292864 TI - Nucleotide sequence and properties of the murine gamma 3 immunoglobulin heavy chain gene switch region: implications for successive C gamma gene switching. AB - During B lymphocyte differentiation, immunoglobulin heavy chain constant region (CH) genes undergo a unique series of DNA recombination events culminating in the CH class switch. CH switch (S) regions are located 2 kb 5' of each CH gene except delta (i.e. mu, gamma 3, gamma 1, gamma 2b, gamma 2a, epsilon and alpha). We describe the structural features of the gamma 3 switch region. Hybridization experiments show that S gamma 3 has remarkable homology to both S mu and other S gamma regions while S mu possesses limited homology to the other S gamma sequences. However, S mu possesses extensive sequence homology with S epsilon and S alpha. The nucleotide sequence of S gamma 3 reveals higher densities of S mu repetitive sequences (GAGCT and GGGGT) and another S region common sequence (YAGGTTG) than observed for S gamma 1, S gamma 2b or S gamma 2a. In addition, the conservation of S mu like repetitive sequences in S gamma regions is correlated with the 5' leads to 3' gamma gene order (i.e. S gamma 3 greater than S gamma 1 greater than S gamma 2b greater than S gamma 2a). A model is presented which suggests that the unique features of S gamma 3 may allow for successive switches from C mu to any C gamma gene. PMID- 6292865 TI - Structure and evolution of the heavy chain from rat immunoglobulin E. AB - The nucleotide sequence of the rat epsilon-chain mRNA has been determined by sequencing cloned cDNA copies of the mRNA. The established sequence covers the coding region, the 3'-non coding region and most of the 5' non-coding region. A comparison with the nucleotide sequence of the human epsilon-chain constant region reveals that C3 and C4 are the most highly conserved domains. The rat epsilon-chain contains a C-terminal decapeptide which is not present in the human counterpart. PMID- 6292866 TI - Analysis of tet operator-TET repressor complexes by thermal denaturation studies. AB - Interaction of the Tn10 encoded TET repressor with the tet operator is studied by thermal denaturation of the specific complexes employing operator containing purified DNA restriction fragments varying in length from 187 bp to 501 bp. Comparison of the melting curves obtained with the free DNA and DNA.repressor complexes revealed a specific stabilisation of the operator containing cooperatively melting segment in multiphasic denaturation curves. Under limiting concentrations of TET repressor the denaturation of the free DNA is observed next to the denaturation of the repressor.DNA complex. Quantitative analysis yields a binding curve with a stoichiometry of four TET repressors per tet operator containing fragment. The denaturation temperature of the complex is almost independent of the ionic strength indicating that the protein component denatures at this temperature. The half life time of the TET repressor.tet operator complex is greater than 100 min under these conditions. The tet operator on the 187 bp fragment is determined to be located between a Xba I and a Sau 3a site by removing base pairs from either end of the fragment and subsequent comparison of the melting curves. It is concluded that the TET repressor recognizes the double stranded rather than a possible cruciform structure of the tet operator. The influence of a regulatory protein on the thermal stability of a genetic control region is discussed with respect to its possible influence on the initiation of transcription. PMID- 6292867 TI - Nucleotide sequence of the 30K protein cistron of cowpea strain of tobacco mosaic virus. AB - The nucleotide sequence of cloned cDNA copies of cowpea strain of tobacco mosaic virus RNA including the 30K protein cistron was determined. The 30K protein cistron was located at residue 676-1,527 from the 3' end of the genomic RNA. The 30K protein was composed of 282 amino acid residues and was basic, similar to the 30K protein of common strain OM. However, homology of the amino acid sequences between the two strains was only 27%. PMID- 6292868 TI - Nucleotide sequence of the fnr gene and primary structure of the Enr protein of Escherichia coli. AB - The nucleotide sequence of a 1.64 kb fragment of E. coli DNA containing the fnr gene (regulatory gene for fumarate and nitrate reduction) was determined using the dideoxy chain termination method. The fnr coding region (750 bp) was identified, and the initiation and termination points of fnr transcription were located by RNA:DNA hybridisation with single-stranded M13 probes. The DNA fragment also contained the 5' end of a separately transcribed gene of unknown function. The deduced molecular weight (27947) of the Fnr protein was in agreement with that of the protein identified by the maxicell procedure, and the primary structure contained regions of homology with several transcriptional regulator proteins. PMID- 6292869 TI - Specific interaction of netropsin, distamycin-3 and analogs with LC duplexes: reversion towards the B form of the 2'-deoxy-.2'-deoxy-2'-fluoro-hybrid duplexes upon specific interaction with netropsin, distamycin-3 and analogs. AB - Binding of the B-form specific ligands netropsin and distamycin-3, -4 and -5 has been used to monitor the presence and/or the inducibility of a B-type structure in various poly-inosinic.poly-cytidilic double stranded polymers with deoxyribose, ribose or 2'-deoxy-2'-fluororibose as sugar on either strand. The efficiency of binding was followed by circular dichroism and further evaluated by the increase in melting temperature of the complexes. The efficient binding of netropsin and distamycins to the hybrid polymer (dIfl)n. (dC)n demonstrated that the fluorine carrying strand may undergo a A to B-type transition reflecting a change of the 2'-deoxy-2'-fluororibose from the 3'-endo to the 1'-exo or 2'-endo pucker. The less efficient binding of the same ligands to the reverse hybrid (dI)n.(dCfl)n showed that the geometry of the pyrimidine strand is the most critical for the specific interaction. Taking into account the recent findings about the regular hydration in the minor groove of the B-type dodecamer dCGCGAATTCGCG in solid-state, the different binding modes observed between the different polymers and antibiotics are explained by differences in their possibilities of hydration. Binding of netropsin to a double stranded deoxypolymer is interpreted as a local replacement of water molecules by netropsin in the minor groove hydration network which is typical of the B-form. PMID- 6292871 TI - The diagnostic application of radiocolloid liver scintigraphy in breast carcinoma. AB - The authors have reviewed their experiences in determining the presence of liver metastases in 103 patients by radiocolloid scanning. The sensitivity of liver scanning proved to be quite low if the presence of focal defects in the distribution of the tracer was chosen as the diagnostic criterion. The inclusion of less restrictive criteria such as liver enlargement or irregular distribution of the tracer, resulted in a higher sensitivity without lowering the predictive value of a negative scan. Using the latter diagnostic criterion, sensitivity, specificity and accuracy were in the range of 90%. Abnormal liver scans are common in patients classified at T3-T4 or N+ and their chances of being "true positive" are high. Conversely, abnormal scans are seldom found in patients classified at T1-T2 or N0 and probabilities of "false positive" results are high. PMID- 6292870 TI - Effect of intron size on splicing efficiency in retroviral transcripts. AB - To study the effect of intron size on splicing efficiency we have varied the size of the avian leukosis virus (ALV) env mRNA intron in a cloned ALV genome. This was accomplished by deletion of ALV sequences or insertion of phage lambda DNA. The effect of these modifications on splicing was analyzed by microinjection of the modified clones into RSV(-) chicken cells. Viral env mRNA when transcribed and properly spliced within these cells complemented the RSV(-) env deficiency leading to the production of focus forming units. Using this assay it was shown that deletion of up to 3.7 kb of the 4.68 kb env intron did not inhibit correct splicing nor did insertion of up to 8 kb of phage lambda DNA prevent splicing. Our results indicate that intron size can be varied over a wide range without preventing splicing. PMID- 6292872 TI - A reinterpretation of phosphatase-N. AB - In order to determine the substrate specificity of the alkaline phosphatase (APase) which appears in murine lymphomas, two substrates were chosen. p Nitrophenyl phosphatase (pNPP), the standard substrate, has an oxygen-phosphorus (O-P) bond. Cysteamine-S-phosphate (CASP) has a sulfur-phosphorus (S-P) bond. It has been reported that murine lymphoma APase does not cleave the S-P bond of CASP. These results were not confirmed. Under all conditions tested, the murine lymphoma APase showed consistent hydrolysis of CASP at approximately one-third the rate of hydrolysis of pNPP. Biochemical characteristics used to confirm this include pH optimum, heat inactivation, kinetics, magnesium activation, L homoarginine inhibition, EDTA inhibition, and activity remaining during stages of partial purification. It is concluded that the murine lymphoma APase shows a preference of pNPP as a substrate but does hydrolyse CASP at a lower rate. The conflict between our laboratory and that of Neumann's may be one of interpretation rather than data. Neumann shows similar data to ours in terms of the 3-fold ratio, but considers all hydrolysis of CASP as representative of 'normal' APase. She uses a formula to calculate the amount of lymphoma APase based on the ratio 1.9 which is the ratio of hydrolysis of pNPP to that of CASP in kidney tissue. Our interpretation would be that the same isozyme is hydrolyzing the S-P bond at one-third the rate that it hydrolyzes the O-P bond. PMID- 6292873 TI - Non-seminomatous germ cell tumors of the testis. Immunohistochemical localization and serum levels of human chorionic gonadotropin (HCG) and pregnancy-specific beta-1 glycoprotein (SP-1); value of SP-1 as a tumor marker. AB - Serum levels of HCG and SP-1 and tissue localization in the primary tumors of these markers were correlated in 41 patients with advanced-stage non-seminomatous germ cell tumors (NSGCT) of the testis. In patients' serum, as well as in the primary tumor tissue, SP-1 positivity was accompanied by a concordant HCG positivity, so that it is concluded that in our patient group SP-1 does not give important additional information with respect to marker positivity not yet provided by HCG. A good correlation was found between serum levels of HCG and SP 1 in advanced-stage disease and the cellular localization of these markers in the primary tumors. Elevated serum SP-1 levels, with concordant elevation of HCG serum levels, were found mainly in cases with a histological diagnosis of choriocarcinoma or an admixture of this component in the primary tumor. Moreover, in cases of choriocarcinoma the presence of SP-1 positive syncytiotrophoblastic cells was very much comparable with the presence of HCG-positive syncytiotrophoblastic cells, whereas in syncytial giant cells associated with embryonal carcinoma HCG-positivity clearly outnumbered SP-1-positivity. This indicates a higher specificity of SP-1 for choriocarcinoma and, therefore, serum levels of this marker may have an important prognostic impact in NSGCT patients with advanced-stage disease. Apart from positivity in syncytiotrophoblastic cells of choriocarcinoma and syncytial giant cells, HCG positivity was incidentally seen in mononuclear embryonal carcinoma cells. PMID- 6292874 TI - Interview with Sergio Stagno: Isolation precautions for patients with cytomegalovirus infection. PMID- 6292876 TI - Concomitant diffuse nodular pulmonary infiltration in an infant with cytomegalovirus infection. PMID- 6292875 TI - IgM to varicella-zoster virus: demonstration in patients with and without clinical zoster. AB - Antibody to varicella-zoster (VZ) virus of the IgM type was detected in sera from 50% of persons with clinical zoster, 67% of asymptomatic varicella immunes with a recent intimate exposure to VZ virus and 22 to 40% of varicella immunes with no symptoms of zoster or known exposure to the virus. No VZ IgM was found in newborn sera or in sera from persons susceptible to varicella, demonstrating specificity of the VA IgM assay. Since development of specific IgM is associated with acute infection, these data suggest that reinfection with VA virus occurs and also that antigenic stimulation due to exposure to endogenous VZ virus may occur. We hypothesize that during reactivation of VZ virus, persons with intact VZ cell mediated immunity (CMI) remain asymptomatic but that those with depressed VZ CMI may develop clinical zoster. These data suggest that there is an unstable relationship between VZ virus and the human host. PMID- 6292877 TI - Drugs for systemic mycoses. PMID- 6292878 TI - Newer agents causing pneumonitis in early infancy. PMID- 6292880 TI - [172 cases of Wilms' tumor in children]. PMID- 6292879 TI - [Radiological symptomatology of primary neoplasms of the ribs]. PMID- 6292881 TI - [Galactography - preliminary report]. PMID- 6292882 TI - [Immunotherapy of lung cancer]. PMID- 6292884 TI - Defining the pathology of endometrial hyperplasia, dysplasia and carcinoma. PMID- 6292883 TI - [Microbiological evaluation of the results of penicillin Hx treatment of urinary tract infection]. PMID- 6292885 TI - Regional variations in the testicular dependence of prolactin binding and its possible relationship to castration-induced involution in rat prostate gland. AB - Prolactin binding sites of ventral, lateral, and dorsal lobes of rat prostate were examined immunohistochemically 1, 2, 4, and 8 days after castration or sham operation. In sham-operated rats each lobe exhibited a distinct pattern of intracellular and intraluminal prolactin binding. A loss in prolactin binding from epithelial cells of ventral prostate, which was visualized postcastration, was quantitated with the aid of an image analyzer and then statistically evaluated. The proportion of ventral prostate epithelial cells devoid of prolactin binding increased from approximately 13% in sham-operated rats to approximately 29% 1 day after castration, and reached a peak level of about 71% 4 days postcastration. No loss of prolactin binding was evident in either lateral or dorsal prostate up to 8 days postcastration. Direct measurement of epithelial cell heights and subsequent statistical evaluation revealed similar regional differences in the rates and extent of prostate involution. Eight days after castration ventral prostate epithelial cell heights decreased by 56% whereas lateral and dorsal lobe epithelial cells heights decreased about 25% and 14%, respectively. The apparent relationship between testicular dependence of prostatic prolactin binding and castration-induced prostatic involution are discussed in terms of possible regional variations in the prolactin-androgen interplay in prostate. PMID- 6292886 TI - Oat cell carcinoma of bronchus presenting as an acute psychiatric illness in young women. PMID- 6292887 TI - [Tracheal resection for tracheal tumors (excluding cancer)]. AB - Tracheal resection was performed in 10 patients with non-malignant tracheal tumors. Clinical, histological, and technical features are discussed, particular emphasis being given to long-term results. These tumors, mostly cylindromas (7 out of 10 cases in this series) are amenable to very wide surgical resection of the resection-anastomosis type. Suggested treatment for cylindromas should include systematic postoperative irradiation, as this considerably improves long term (10 and 15 years) results. PMID- 6292888 TI - [Branchiomeric mediastinal chemodectoma: a case report and literature review]. AB - Among non-chromaffin paragangliomas, the so-called aorticopulmonary or branchiomeric mediastinal chemodectomas are very rare tumors. A case is reported, and 57 similar cases described in the literature are reviewed. As recommended by Olson, a separate group was instituted of 21 cases of the so-called posterior mediastinal or aorticosympathetic group of the costovertebral groove. In principle, aorticopulmonary chemodectomas are non-functional, and in 9 out of 10 cases follow a slow and benign course extending over many, or even tens of years. In contrast to carotid and jugular paragangliomas they are rarely multifocal: when this is the case it is difficult to confirm the presence of metastases, this being however the most reliable criterion of their malignancy. The fortuitous discovery on a radiography of a tumor of the anterior and middle compartments of the upper mediastinum should invoke the presence of an aorticopulmonary chemodectoma, and lead to arteriography of the aortic arch region. Diagnosis is made essentially by pathological examination. Treatment is exclusively by surgical excision, and this was complete in nearly half of the cases treated. Prognosis is not hopeless after partial removal, however, and a relatively comfortable survival can be obtained extending over many years. PMID- 6292889 TI - Visceral leishmaniasis in europe. PMID- 6292890 TI - Chemical and biochemical studies in fetuses affected with Nieman-Pick disease type A. AB - Chemical and biochemical studies were performed on two unrelated fetuses affected with Niemann-Pick disease type A, following abortion at about the 19th week of gestation. Abortion was performed as a consequence of previous findings, in amniotic fluid cell cultures, that sphingomyelinase activity was completely absent. Phospholipid analyses of various organs of the fetuses, spleen and liver were the organs mostly affected. Interestingly enough considerable accumulation of sphingomyelin was found in the placenta. The brain was the only organ in which sphingomyelin storage could not be proved. In addition to sphingomyelin a slight accumulation of cholesterol was noticed. Deficiency of sphingomyelinase activity measured at pH 5.0 was the general characteristic of the affected tissues. It is concluded that the accumulation of sphingomyelin in various organs throughout the body of fetuses affected with Niemann-Pick disease is suggestive of the essential role of the enzyme sphingomyelinase and its biochemical maturation, even during the early stages of gestation. PMID- 6292891 TI - Removal of superoxide dismutase activity from cytochrome C. AB - Commercially available cytochrome c contains sufficient superoxide dismutase activity to reduce its sensitivity in superoxide anion detection. A single passage through a column of Sephadex G-50 removes the superoxide dismutase, and appreciably increased the ability to cytochrome c to detect superoxide. PMID- 6292892 TI - [Hormonal and immunological status of children with thymus hyperplasia]. AB - The state of the adrenocortical system, cellular and humoral immunity was studied in 152 infants, suffering from thymomegaly, associated with virus-bacterial pneumonia. The results obtained allow one of to consider the hormonal and immunological state of the patients with thymomegaly to be inhibited and the adrenocortical hypofunction to be secondary. Therefore, the infants with thymomegaly should be separated into a special "risk" group, according to their immunodeficient states, respiratory allergies, acute and chronic adrenocortical deficiency. PMID- 6292893 TI - Nucleotide sequence of the ilvB promoter-regulatory region: a biosynthetic operon controlled by attenuation and cyclic AMP. AB - The DNA sequence of the promoter-regulatory region of the ilvB operon of Escherichia coli was determined. This region encodes a potential leader polypeptide containing 32 amino acids, 12 of which are the regulatory amino acids valine and leucine. Approximately 50 residues downstream from the coding region for the potential leader peptide is a site for terminating transcription. In vitro transcription experiments show that transcription terminates at this site and produces a leader mRNA of approximately equal to 188 nucleotides. A model for the multivalent regulation of this operon by valyl- and leucyl-tRNA is proposed on the basis of the mutually exclusive formation of five strong stem-and-loop structures in the leader mRNA. In addition, the -35 and -70 regions of this sequence show close structural homologies to areas in cyclic AMP receptor protein (CRP)-dependent promoters reported to be important for CRP function. In vitro transcription from the ilvB promoter was greatly increased by cyclic AMP and CRP. Taken together, these data strongly suggest that the ilvB biosynthetic operon is negatively controlled by multivalent transcription termination and is positively regulated by cyclic AMP and CRP. PMID- 6292894 TI - Receptor-mediated endocytosis of transferrin and the uptake of fe in K562 cells: identification of a nonlysosomal acidic compartment. AB - At physiological temperature, the Fe-carrier transferrin is taken up by K562 human erythroleukemia cells through receptor-mediated endocytosis. Both ligand (now minus Fe) and receptor recycle back to the cell surface where the receptor is rapidly reutilized. After endocytosis, transferrin becomes transiently lodged within an acidic compartment inside the cell, as judged by the changed spectral characteristics and quantum yield of fluorescein isothiocyanate-labeled transferrin that is cell-associated at 37 degrees C. Upon binding to transferrin, anti-fluorescein antibody strongly quenches the emission of the fluorescein labeled residues on the protein and is used to assess whether the transferrin is at the cell surface (incubation at 0 degrees C) or mainly internalized into the cell (incubation at 37 degrees C). Using Percoll gradient fractionation of postnuclear supernatants, we show that the acidic compartment is not the lysosomal compartment. PMID- 6292895 TI - Corticotropin-releasing factor stimulates phospholipid methylation and corticotropin secretion in mouse pituitary tumor cells. AB - The 41-residue synthetic ovine corticotropin-releasing factor (CRF; corticoliberin) has been shown to stimulate release of corticotropin (adrenocorticotropic hormone; ACTH) and beta-endorphin from AtT-20/D16-16 mouse pituitary tumor cells. Phospholipid methylation of phosphatidylethanolamine to phosphatidylcholine with S-adenosylmethionine as methyl donor has been suggested as a possible membrane transduction mechanism for some receptor-induced events. CRF increased phospholipid methylation in pituitary tumor cells at concentrations that also stimulated immunoreactive ACTH secretion, and both processes increased linearly and in parallel with time. The methionine sulfoxide derivative of CRF was less potent than CRF was in stimulating both phospholipid methylation and hormone secretion, and the COOH-terminal free acid analogue of CRF had no effect on either process. CRF-induced increases in phospholipid methylation and ACTH secretion were reduced when cells were treated with the phospholipid methyltransferase inhibitors 3-deazaadenosine and L-homocysteine thiolactone. These CRF-stimulated effects were also blocked by the glucocorticoid dexamethasone. It is suggested that phospholipid methylation may be a CRF receptor-mediated event associated with ACTH release in pituitary tumor cells. PMID- 6292896 TI - IgA interaction with the asialoglycoprotein receptor. AB - IgA present in normal human serum reacts with the hepatic receptor specific for asialoglycoproteins as demonstrated by inhibition of receptor-mediated erythroagglutination. Inhibition is reversibly abolished by the oxidation of the galactose or N-acetylgalactosamine residues of IgA with galactose oxidase. The site of receptor recognition appears to be the O-glycosidically linked oligosaccharides present on the hinge region of the IgAI subtype of IgA. The demonstration of a specific binding, in vitro, of IgA by the hepatic receptor suggests that the uptake of polymeric IgA by the liver in vivo may be mediated by this reaction. PMID- 6292897 TI - In vitro assembly of the nonglycosylated membrane protein (M) of Sendai virus. AB - The nonglycosylated membrane protein (M) of Sendai virus was purified from virions and conditions were found under which the protein assembled in vitro into three types of ordered structures: narrow tubes, wide tubes, and sheets. These structures were examined by high resolution electron microscopy by using negative staining and metal shadowing techniques. The tubes and sheets are formed from strands 7.2 nm wide that are composed of annular subunits. The wide tubes appear to be formed by the rolling of a sheet into a cylinder in which the 7.2-nm strands are inclined with a pitch of 26-33 degrees and have a left-handed orientation. In addition to the strong reflections corresponding to the 7.2-nm spacings generated by the strands, optical diffraction patterns also showed weak reflections that could be indexed on a lattice corresponding to real-space lattice constants of 7.6 nm and 5.3 nm, with an included angle of 71 degrees. The dimensions and arrangements of these structures formed in vitro are strikingly similar to those of ordered arrays of particles found by others to be associated with the inner surface of the plasma membrane of infected cells. The results support the concept that ordered arrays of M protein, similar to those assembled in vitro, are involved in the assembly of the virus particle by budding from the cell membrane and that they provide specific recognition sites for the viral nucleocapsid at the cytoplasmic surface of the plasma membrane. PMID- 6292898 TI - Microinjection of monoclonal antibody to protein p53 inhibits serum-induced DNA synthesis in 3T3 cells. AB - Monoclonal antibody directed against the transformation-related protein p53 was microinjected manually into the nuclei of quiescent Swiss 3T3 mouse cells. The cells were subsequently stimulated with 10% fetal calf serum. Microinjection of p53 antibody at or around the time of serum stimulation clearly inhibited the subsequent entry of Swiss 3T3 cells into the S phase of the cell cycle. p53 antibody had no effect on serum-stimulated DNA synthesis when it was microinjected 4 hr or later after serum stimulation. Monoclonal antibody to an unrelated antigen, Lyt-2.2, had no effect on serum-stimulated DNA synthesis regardless of the time it was microinjected. Under similar experimental conditions, p53 antibody had no effect on simian virus 40- or adenovirus 2 induced DNA synthesis. These experiments add strength to the suggestion that p53 is involved in the regulation of cell proliferation. PMID- 6292899 TI - Difference in saturable binding of low density lipoprotein to liver membranes from normocholesterolemic subjects and patients with heterozygous familial hypercholesterolemia. AB - To investigate the possible role of the low density lipoprotein (LDL) receptor in the catabolism of LDL by the human liver, the binding of 125I-labeled LDL to membrane fractions prepared from human liver biopsies was determined. Biopsy samples taken for routine histology were obtained from seven patients with heterozygous familial hypercholesterolemia, one with non-familial hypercholesterolemia, and seven normocholesterolemic subjects. LDL was bound by the membranes from normal subjects in a saturable manner that was inhibited by 56% in the presence of excess LDL. Binding of LDL was also inhibited by modification of the lipoproteins with 1,2-cyclohexanedione. The amount of 125I labeled LDL bound to membranes from familial hypercholesterolemic livers that could be displaced with excess LDL was significantly less than that bound by normocholesterolemic membranes. These observations suggest that LDL receptors are expressed in normal human liver and are defective in the livers of familial hypercholesterolemic patients. PMID- 6292900 TI - Antibiotic-DNA interactions: intermolecular nuclear Overhauser effects in the netropsin-d(C-G-C-G-A-A-T-T-C-G-C-G) complex in solution. AB - The proton markers located in the minor groove of the d(C-G-C-G-A-A-T-T-C-G-C-G) duplex and its netropsin complex have been assigned from measurements of intramolecular nuclear Overhauser effects (NOEs) between exchangeable imino protons and nonexchangeable base protons on the same and adjacent base pairs. Several points of contact between the concave face of the antibiotic and the minor groove d(A-A-T-T) tetranucleotide segment of the dodecanucleotide duplex have been established based on intermolecular NOE effects between the pyrrole ring and side-chain methylene protons of netropsin and the adenosine H-2 protons of dA X dT base pairs in the center of the duplex. These NOE measurements provide a powerful method for differentiating between minor and major groove contacts in ligand-DNA complexes in solution. A model for netropsin interaction at dA X dT sites on duplex DNA is proposed. PMID- 6292901 TI - Host-specific activation of transcription by tandem repeats from simian virus 40 and Moloney murine sarcoma virus. AB - The simian virus (SV40) 72-base pair (bp) tandem repeated sequences have recently been shown to function as activators or enhancers of early viral transcription. A recombinant viral genome was recently constructed by inserting 72-bp tandem repeats from the Moloney murine sarcoma virus (MSV) in place of the 72-bp repeats of SV40. Although this genome replicates in monkey kidney cells, its rate of large tumor antigen expression and replication is considerably slower than that of wild-type SV40. In mouse cells, however, equivalent levels of large tumor antigen appear to be expressed from both wild-type and recombinant genomes, suggesting a relationship between the level of enhancer activity and the host cell. To confirm this observation, we have applied a sensitive quantitative assay for gene expression based on the conversion of chloramphenicol to its acetylated forms. The gene encoding the enzymatic function chloramphenicol acetyltransferase was inserted into two vectors in which the enhancer sequences from SV40 or MSV were placed adjacent to the early SV40 promoter. The SV40 tandem repeats appear to activate gene expression to significantly higher levels in monkey kidney cells, but the MSV repeats are more active in two lines of mouse cells. These findings suggest that the tandem repeat elements may interact with host-specific molecules and, furthermore, may constitute one of the elements determining the host range of these eukaryotic viruses. PMID- 6292902 TI - Sodium nitroprusside-induced protein phosphorylation in intact rat aorta is mimicked by 8-bromo cyclic GMP. AB - The effects of sodium nitroprusside, 8-bromo cyclic GMP, 8-bromoguanosine 5' monophosphate, 8-bromo cyclic AMP, dibutyryl cyclic AMP, and isoproterenol on incorporation of (32)P into proteins in intact rat thoracic aorta were studied. Aortas were incubated in [(32)P]orthophosphate in order to label endogenous adenosine triphosphate. Agents were then added for various times and the tissues were homogenized and fractionated (100,000 x g for 60 min) into soluble and particulate fractions. Soluble and particulate fractions were subjected to isoelectric focusing followed by sodium dodecyl sulfate/polyacrylamide gel electrophoresis and autoradiographs were made. Nitroprusside induced a concentration-dependent increase in incorporation of (32)P into nine proteins and a decrease in (32)P incorporation into two proteins. Some of these proteins appeared in both the soluble and particulate fractions of homogenates; others appeared only in the soluble fraction. The pattern of (32)P incorporation was identical after 2- or 15-min exposure to nitroprusside and was mimicked by exposure to 50-500 muM 8-bromo cyclic GMP. 8-Bromoguanosine 5'-monophosphate did not alter (32)P incorporation. Dibutyryl cyclic AMP at 50 muM had no effect upon (32)P incorporation whereas a higher concentration (0.5 mM) caused increased or decreased (32)P incorporation into some, but not all, of the same proteins. 8 Bromo cyclic AMP (5 mM) produced only small changes in (32)P incorporation. The pattern of (32)P incorporation induced by a relatively high concentration of isoproterenol 0.1 mM was similar but not identical to that seen with 0.5 mM dibutyryl cyclic AMP. The present study indicates that the incorporation of (32)P into endogenous proteins of intact rat aorta can be regulated by nitroprusside. These effects can be mimicked by cyclic GMP analogues and only partially by cyclic AMP analogues or isoproterenol. Presumably, these effects of nitroprusside are mediated through a cyclic GMP-dependent process (protein kinase or phosphatase) which may play a role in the relaxant properties of nitroprusside and cyclic GMP. PMID- 6292903 TI - Post-transcriptional nucleotide addition is responsible for the formation of the 5' terminus of histidine tRNA. AB - All sequenced histidine tRNAs have one additional nucleotide at the 5' end when compared to other tRNA species. Sequence analysis of histidine tRNA genes from Drosophila melanogaster and Schizosaccharomyces pombe showed that the terminal guanylate residue of the mature tRNAs is not encoded by the genes. Analysis of the products from in vitro transcription of these genes in extracts from Drosophila Kc cells demonstrated that the 5'-terminal nucleotide present in the mature tRNA is added post-transcriptionally. The addition reaction requires ATP. A portion of the mature tRNAs are then modified at the 5'-terminal pG. Analysis of the RNA species formed during the in vitro maturation of the Drosophila histidine tRNA primary transcript uncovered the following maturation scheme: (i) the primary transcript is processed by RNase P at the 5' end to form an intermediate precursor; (ii) the 3'-flanking sequence is endonucleolytically removed, and a guanylate moiety is added to the 5' end to form mature-sized histidine tRNA; and (iii) a fraction of the 5'-terminal guanylate residues then undergoes modification. In contrast to the capping of eukaryotic mRNA, the guanylate addition to histidine tRNA results in the formation of a (3'-5') phosphodiester bond. There are no precedents for the post-transcriptional addition of nucleotides (in phosphodiester linkage) to the 5' end of RNA precursors. PMID- 6292904 TI - Characterization of beta-endorphin binding protein (receptor) from rat brain membranes. AB - Rat brain membranes (RBM) bind beta-endorphin with high affinity and specificity. We report herein the identification of a high molecular weight beta-endorphin complex (receptor) in extracts of RBM preincubated with tritiated beta-endorphin, by using the zwitterionic detergent 3-[(3-cholamidopropyl)dimethylammonio]-1 propanesulfonate (CHAPS). Upon isoelectric focusing, this complex gives a single peak with an isoelectric point (+/- SEM) of 4.50 +/- 0.06. Sucrose density gradient experiments in H2O and 2H2O yield effective partial specific volume (v = 0.814 cm3/g and sedimentation constant s20,w = 15.6 S. Gel filtration yields an estimate of the hydrodynamic radius of 73 A. The corresponding frictional ratio of 1.12 is consistent with an elliptical spheroid with an axial ratio of 3.1-3.2. The molecular mass of the complex is estimated to be 690,000 daltons. The v of the complex is greater than that of CHAPS or most globular proteins. PMID- 6292905 TI - Cloning and characterization of different human sequences related to the onc gene (v-myc) of avian myelocytomatosis virus (MC29). AB - We have studied the genomic organization of human cellular sequences (c-myc) homologous to the transforming gene (v-myc) of avian myelocytomatosis virus (MC29). Southern blotting experiments using v-myc probes showed that several fragments of the human genome contain sequences related to the central part of v myc but only few of them are homologous to the 3' portion of the viral gene. Several recombinant phages which represent different regions of the genome containing c-myc-related sequences were isolated from a human DNA library. Two clones (lambda-LMC-12 and -41) overlap over approximately 17 kilobases of DNA where a sequence homologous to that of the entire v-myc is present. Restriction mapping experiments and heteroduplex analysis show that c-myc sequences of this locus are interrupted by one intron, suggesting that lambda-LMC-12 and -41 contain the complete functional c-myc gene. Three other clones (lambda-LMC-3, -4, and -26) do not overlap and contain sequences related to only approximately 0.3 kilobase of v-myc but lack 5' and 3' portions of the gene. These sequences are not interrupted by introns and are more divergent from v-myc than is the complete gene, suggesting that they may represent either pseudogenes or parts of distantly related genes. PMID- 6292906 TI - Regulation of double-stranded RNA-activated eukaryotic initiation factor 2 alpha kinase by type 2 protein phosphatase in reticulocyte lysates. AB - Protein synthesis initiation in reticulocyte lysates is inhibited by low concentrations (1-20 ng/ml) of double-stranded RNA (ds RNA) due to the activation of a ds RNA-dependent cAMP-independent protein kinase (ds I) that phosphorylates the alpha subunit of the eukaryotic initiation factor eIF-2. In lysates, ds I is present in the latent inactive form and is associated with the ribosome complement. Latent ds I is solubilized by extraction with high-salt buffers and can be purified in its latent form. Activation of purified latent ds I requires ds RNA and ATP and is accompanied by the ds RNA-dependent autophosphorylation of a polypeptide doublet of 70,000 and 72,000 daltons ("70k/72k"), which represent different phosphorylated states of the same polypeptide. These are phosphorylated in the sequence 70k-->72k; increased phosphorylation of 72k is associated with increased ds I activation. Lysates (or Sepharose 6B ribosomes) treated with ds RNA display a similar ds I phosphoprotein profile, and this is accompanied by the phosphorylation of endogenous eIF-2alpha (38,000 daltons). Delayed (32)P pulses in ds RNA-inhibited lysates indicate that the phosphates on ds I and eIF-2alpha turn over. Under defined conditions, activated ds I in lysates is selectively dephosphorylated by endogenous protein phosphatase(s), and this is accompanied by the dephosphorylation of eIF-2alpha. Similarly, purified activated ds I is rapidly dephosphorylated by unfractionated lysate protein phosphatase(s) and by type 2 protein phosphatase but not by type 1 protein phosphatase. The dephosphorylation of ds I occurs in the sequence 72k-->70k and is correlated with ds I inactivation. The heat-stable protein phosphatase inhibitor-2, which selectively blocks type 1 protein phosphatase, does not significantly affect the dephosphorylation of ds I by type 2 protein phosphatase or by unfractionated lysate phosphatases. The data support the conclusion that a ds I phosphatase activity with type 2 characteristics is involved in the regulation of ds I activity. PMID- 6292907 TI - Hormonal regulation of protein synthesis, secretion, and phosphorylation in cultured rat Sertoli cells. AB - The accumulation of two polypeptides, SCm1 and SCm2, in the medium of Sertoli cell cultures is enhanced by follicle-stimulating hormone (FSH) but is unaffected by either the cAMP analog, N6,O2'-dibutyrl cAMP or luteinizing hormone. The assigned molecular weights of SCm1 and SCm2 differ from those of androgen-binding protein subunits or any other previously identified Sertoli cell secretory product. Incubation of Sertoli cell cultures with either FSH or N6,O2'-dibutyryl cAMP also stimulates the incorporation of [35S]methionine into two intracellular polypeptides, SCc1 and SCc2. In addition, the phosphorylation of three intracellular polypeptides, SCc3, SCc4, and SCc5, is intensified when Sertoli cell cultures are treated with either FSH or N6,O2'-dibutyryl cAMP. Based on these results and on previous work, we conclude that (i) SCm1 and SCm2 may, like androgen-binding protein, be secreted by Sertoli cells and function extracellularly while SCc1 and SCc2 are involved in FSH-dependent intracellular activity; (ii) SCc3, SCc4, and SCc5 are possible substrates for FSH-stimulated, cAMP-dependent protein kinase activity; and (iii) SCc5 is an isoelectric variant of vimentin-type intermediate filament protein presumably involved in FSH- and N6,O2'-dibutyryl cAMP-induced Sertoli cell shape changes. PMID- 6292908 TI - Evidence for multiple origins of the beta E-globin gene in Southeast Asia. AB - To investigate whether recurrent mutation has contributed to the high frequency of the beta E-globin gene in Southeast Asia, we used the haplotypes at three polymorphic restriction sites within and to the 3' side of the beta-globin gene to predict the framework of 23 beta E-globin genes. These haplotypes suggested that beta E-globin genes are present in two different beta-globin gene frameworks. DNA sequence determination of one gene representing each framework demonstrated that the same mutation (GAG leads to AAG at codon 26) was present in both frameworks. Moreover, the frameworks differed at three nucleotide positions known to be polymorphic in Mediterraneans. These polymorphic sites are located 70 nucleotides to the 5' side of the beta E mutation and 382 and 1032 nucleotides to the 3' side of it. The existence of the beta E mutation in these two beta-globin gene frameworks can be explained by (i) recurrent mutation giving rise to beta E globin, (ii) a double crossing-over event, or (iii) two single crossing-over events. Mathematical analysis suggests that the first alternative, recurrent mutation of G leads to A at the first nucleotide of codon 26, is most likely. PMID- 6292909 TI - Expression of herpes simplex virus glycoprotein C from a DNA fragment inserted into the thymidine kinase gene of this virus. AB - Previous reports have described mutants of herpes simplex virus type 1 that fail to produce or accumulate one of the major glycoproteins, glycoprotein C (gC). This defect is not lethal in cell culture, has been associated with the syncytial plaque morphology of some mutants, and may result from mutations that map to a region on the genome noncontiguous with the structural gene for gC. To investigate the conditions required for, and consequences of, gC expression in a specific genetic background, we have inserted a wild-type allele of the gC gene into the thymidine kinase gene (tk) of a gC- fusion-inducing viral mutant, strain MP. This was accomplished by identifying cloned viral DNA fragments homologous to gC mRNA, inserting the appropriate fragments into the viral tk cloned in pBR322, and then cotransfecting cells with the recombinant plasmids and DNA from strain MP, for selection of insertional TK- mutants. All TK- mutants containing insertions of appropriate sequences (in either orientation) into tk were found to express gC while maintaining the syncytial plaque morphology of strain MP. Elimination of the insertion from one of the TK- mutants was accompanied by loss of ability to produce gC. Our results permit more precise mapping of the DNA sequence encoding gC, to a subfragment of Sal I fragment R (map coordinates 0.620 0.640) and indicate also that promoter sequences for the gC gene may be located in this fragment. Moreover, we can conclude that the previously described regulatory mutation of strain MP does not prevent expression of gC from the DNA inserted into its gene tk and that the syncytial phenotype of MP cannot be due solely to absence of gC. PMID- 6292910 TI - Human type I procollagen genes are located on different chromosomes. AB - A recombinant plasmid containing sequences complementary to human pro-alpha l(I) collagen mRNA was used for the chromosomal assignment of the pro-alpha l(I) collagen gene. Restriction endonuclease analysis of DNA from mouse-human and Chinese hamster-human somatic cell hybrids revealed cosegregation with human chromosome 17. Hybrids containing derivative chromosomes with a t(2;17)(q14;q21) translocation showed cosegregation of the pro-alpha l(I) gene with the segment 17q21 leads to qter. In situ hybridization on human metaphasic chromosomes confirmed this conclusion. PMID- 6292911 TI - Human hybridomas constructed with antigen-specific Epstein-Barr virus-transformed cell lines. AB - A 6-thioguanine-resistant, human lymphoblastoid B-cell line (GM1500 6TG A-11; IgG secreting) was mutagen-treated with low-level gamma-irradiation and selected for ouabain resistance. One line showing 10,000-fold higher drug resistance, designated KR-4, was fused with an Epstein-Barr virus-transformed, cloned, B lymphocyte cell line (B6) producing antitetanus toxoid (TT) antibody (IgM), and the hybrids were selected in hypoxanthine/aminopterin/thymidine medium containing 10 microM ouabain. Surviving cells, which arose at an optimal frequency of 10( 5), were subcloned by limiting dilution and screened for anti-TT production. Out of 395 final subclones, 372 were found positive for anti-TT, and seven that were selected for further study secreted specific antibody (IgM, kappa chain) at a maximum concentration of 3-6 micrograms/ml. The differential rate of anti-TT production during the logarithmic phase of cell growth was 15-fold higher in the hybridomas than in the original B6 line. The hybrid nature of the clones was confirmed by karyotype analysis, histocompatibility antigen typing, and expression of secreted and membrane-bound Ig classes. Biosynthetic labeling of the cells revealed that all hybrids secreted both IgM and IgG but that only the IgM class had specificity for TT. Because Epstein-Barr virus is a polyclonal B lymphocyte activator, the technique we applied here may be useful for increasing the recovery of rare antigen-specific B cells in the peripheral blood and for improving the frequency and stability of hybridomas secreting a given antibody. PMID- 6292912 TI - Monoclonal antibodies to the thyrotropin receptor: stimulating and blocking antibodies derived from the lymphocytes of patients with Graves disease. AB - Human monoclonal antibodies have been generated from heterohybridomas obtained by fusing mouse myeloma cells with peripheral lymphocytes from patients with active Graves disease. This report characterizes four antibodies as presumptive thyrotropin receptor antibodies because they specifically inhibit thyrotropin binding and competitively inhibit thyrotropin-induced cAMP levels in human thyroid cells. Two of these antibodies, 208F7 and 206H3, are representative of autoimmune stimulators in Graves disease sera because they stimulate thyroid function in all assays, including the mouse bioassay; their ability to inhibit thyrotropin-induced cAMP increases in thyroid cells competitively is complemented by more than additive agonism at low (10 pM) thyrotropin concentrations. These stimulating antibodies interact more potently with human thyroid ganglioside preparations than with bovine thyroid or brain gangliosides; in contrast, they are poor inhibitors of 125I-labeled thyrotropin binding to liposomes containing the glycoprotein component of the human thyrotropin receptor. Antibodies 129H8 and 122G3 appear to be representative of inhibiting or "blocking" antibodies in Graves disease sera. Thus they have no intrinsic stimulatory action in assays of thyroid function but rather inhibit thyrotropin activity in the assays tested. These two antibodies do not react with human thyroid gangliosides but are strong inhibitors of thyrotropin binding to liposomes containing the high-affinity glycoprotein component from human, bovine, and rat thyroid membranes. The data unequivocally establish the pluritopic nature of the immunoglobulins in Graves disease and relate individual components or determinants of the thyrotropin receptor structure with specific autoimmune immunoglobulins. PMID- 6292914 TI - Initial synaptic transmission at the growth cone in Xenopus nerve-muscle cultures. AB - The excellent visibility of cultured cells allows the early events during formation of the neuromuscular junction to be suitably studied. It has been shown in various culture systems that synaptic transmission occurs early after nerve muscle contact. Early synaptic potentials are small in amplitude and slow in time course reflecting a low acetylcholine receptor density at the site of nerve contact. Acetylcholine receptors accumulate later at the contact region. We have examined initial synaptic transmission at the growth cone-muscle contact in Xenopus nerve-muscle cultures. The approaching growth cone was observed under a phase-contrast microscope while the membrane potential of its target muscle cell was continuously monitored by using an intracellular microelectrode. The innervating neuron was stimulated extracellularly at the cell body. No synaptic potential was evoked when the growth cone was contacting the muscle only at the tip of filopodia. However, as soon as the main portion of the growth cone contacted the muscle membrane, nerve-evoked synaptic potentials were detected after stimulation of the nerve. This immediate appearance of synaptic potentials raises the possibility that acetylcholine could be released at the growth cone even prior to contact with muscle cells. As the area of contact enlarged during the observation period the amplitude of end-plate potentials also increased. Spontaneous synaptic potentials (miniature end-plate potentials) were rarely observed in these early growth cone-muscle contacts. Although there were several inherent difficulties, quantal analysis of the end-plate potentials was attempted by using binomial statistics. This analysis suggests that nerve-evoked transmitter release at the growth cone occurs in a quantal fashion. PMID- 6292913 TI - Immunocytochemical localization of sodium channel distributions in the excitable membranes of Electrophorus electricus. AB - The tetrodotoxin binding protein, a major component of the Na+ channel, has been purified from the electric organ of the South American eel Electrophorus electricus. Antibodies to this protein were raised in rabbits and their specificity was demonstrated by a highly sensitive radioimmunoassay and by immunoprecipitation procedures. These antibodies were used to examine the distribution of the binding protein in the eel electroplax membranes and along myelinated nerve axons. The distribution of the antigen was determined by using the peroxidase-antiperoxidase technique at both the light and electron microscopic levels. In the electrocytes of the electric organ, only the innervated face showed staining in experimental material. The stained regions of electroplax plasmalemma included the caveolae of the innervated surface while caveolae of the non-innervated surface did not stain. Thus, the innervated surface including caveolae exclusively contains the Na+ channels. Along myelinated axons, staining was limited to the nodal zone of the node of Ranvier. The paranodal and internodal zones did not stain for the binding protein. Limited diffusion of primary IgG and subsequent reactants into the paranodal and internodal sites was eliminated as a possible source of focal staining at nodes because mechanically demyelinated preparations also exhibited focal nodal staining. Thus, this tetrodotoxin binding protein component of the Na+ channel is located solely within the nodal zone of the node of Ranvier. PMID- 6292915 TI - Modification of single Na+ channels by batrachotoxin. AB - The modifications in the properties of voltage-gated Na+ channels caused by batrachotoxin were studied by using the patch clamp method for measuring single channel currents from excised membranes of N1E-115 neuroblastoma cells. The toxin modified open state of the Na+ channel has a decreased conductance in comparison to that of normal Na+ channels. The lifetime of the modified open state is drastically prolonged, and channels now continue to open during a maintained depolarization so that the probability of a channel being open becomes constant. Modified and normal open states of Na+ channels coexist in batrachotoxin-exposed membrane patches. Unlike the normal condition, Na+ channels exposed to batrachotoxin open spontaneously at large negative potentials. These spontaneous openings apparently cause the toxin-induced increase in Na+ permeability which, in turn, causes membrane depolarization. PMID- 6292916 TI - Cyclic nucleotides and growth regulation in BHK cells. PMID- 6292917 TI - Effect of 3-aminopropanesulfonic acid and allylglycine on prolactin release in male rats. PMID- 6292918 TI - Varicella-zoster virus fails to induce immunoglobulin G Fc receptors in infected human cells. PMID- 6292919 TI - Serum phosphate abnormalities in sickle cell anemia. PMID- 6292920 TI - Localization of radiolabeled antibody in SVT2 tumor increases with immunosuppression of the host. PMID- 6292921 TI - Immune enhancement of viral infection. PMID- 6292922 TI - Histopathological aspects of renal tumors in children. PMID- 6292924 TI - Complications and sequelae of the treatment of Wilms' tumor. PMID- 6292923 TI - Chemotherapy in Wilms' tumor--indications and early side effects. PMID- 6292925 TI - Wilms' tumor updated: evolution of ideas and trends. PMID- 6292926 TI - Preoperative chemotherapy in Wilms' tumour. Results of clinical trials and studies on nephroblastomas conducted by the International Society of Paediatric Oncology (SIOP). PMID- 6292927 TI - Wilms' tumor: histologic grading as a prognostic factor in tumors beyond 10cm in diameter. AB - The progressive proportion of 25-50 and 75% of undifferentiated cells in the Wilms' tumor mass generate three grading categories: well differentiated (WD); the moderate differentiated (MD) and the poorly differentiated (PD). The Urology Department of the Instituto Nacional de Enfermedades Neoplasicas (Lima, Peru) has studied 60 selected cases registered at its Biostatistics Department. Only 6.9% were well differentiated; 69% were of the poorly differentiated type. In 58 patients our survival rate at two years after diagnosis is higher (39.53%) when the patients are over 2 years of age; while under 2 years of age this survival rate is 26.67%. In a group of 46 patients, the survival rate at two years after initial diagnosis is 44% with a tumor size between 10-14 cm; the survival rate drops to 39.3% when the tumor size is over 15 cm in diameter. There is a linear increasing scale of survival rate for the well differentiated (25%), moderate differentiated (27.3%) and for the patients bearing poorly differentiated types of tumors (48.4%). Further studies are encouraged in order to learn the reasons for this correlation. PMID- 6292928 TI - An experimental Wilms' tumor. PMID- 6292929 TI - Surgical aspects of simultaneous bilateral Wilms' tumors. PMID- 6292930 TI - Late-recurring Wilms' tumors. PMID- 6292931 TI - The treatment of Wilms' tumor in 1982 the status of the art. PMID- 6292932 TI - Aetiology of renal cancer. PMID- 6292933 TI - Wilms' tumor: genetic aspects and etiology. PMID- 6292934 TI - Evaluation of nephroblastomas. PMID- 6292935 TI - Computerized tomography and nephroblastomas. PMID- 6292936 TI - Evaluation of unusual nephroblastomas and differential diagnosis. PMID- 6292937 TI - Preoperative diagnosis of Wilms' tumor. PMID- 6292938 TI - Surgery of Wilms' tumor. PMID- 6292939 TI - TMM description and staging of Wilms' tumors. Prognostic factors. PMID- 6292940 TI - Radiation therapy of Wilms' tumor. Methods and immediate complications. PMID- 6292941 TI - Interaction of steroids and growth factors with the plasma membrane in the induction of oocyte maturation in Xenopus laevis. PMID- 6292942 TI - Intracellular calcium and fertilization: role of the cation and regulation of intracellular calcium levels. PMID- 6292943 TI - Cation transport and growth control in neuroblastoma cells in culture. PMID- 6292944 TI - Synapse reformation and repression in muscle reinnervation: an evaluation of endogenous and exogenous influences on nerve regeneration. PMID- 6292945 TI - Gangliosides and neuronal cell differentiation. PMID- 6292946 TI - A polypeptide involved in membrane transport of substrate for glucose-6 phosphohydrolase. PMID- 6292947 TI - Molecular steps in the action and regulation of epidermal growth factor: a cellular mitogen. PMID- 6292948 TI - Thyroid cell polarization in culture and the expression of specialized functions. PMID- 6292949 TI - The role of receptor-mediated endocytosis in iron metabolism. PMID- 6292950 TI - Biosynthesis of the catalytic subunit of (Na+, K+)-ATPase in toad kidney and toad bladder epithelial cells. PMID- 6292951 TI - The entry of enveloped viruses into an epithelial cell line. PMID- 6292952 TI - Regulation by turnover of Na,K-ATPase in HeLa cells. AB - As in most if not all animal cells, HeLa Na,K-ATPase is an essential enzyme of the cell surface. The three ways, referred to in the Introduction, in which it is regulated may be summarized as follows: 1) The activity of the enzyme under normal conditions responds almost linearly to small perturbations in internal (Na+); this is short-term regulation. 2) In the normal steady state, the enzyme is one of the rapidly turning over components of the cell surface; this is part of long-term regulation. The functional importance of turnover to cell homeostasis depends on the reversibility of any event that may inactive the enzyme. Thus turnover is an important, but not the only, means of recovery from ouabain intoxication, and for an inactivating ligand that dissociates more readily than ouabain turnover may be relatively unimportant. Conversely, turnover may be the only means of recovery from thermal inactivation at physiological temperatures. 3) Under conditions of prolonged stress, turnover itself may be modulated so as to enhance the number of active enzymes on the cell surface. Regulation by turnover has the advantage of permitting a prompt response to a changing cell environment, whereas regulation by synthesis would introduce a delay in response at least equal to the transit time. Broadman et al [1974] have suggested that the signal for the increase in Na,K-ATPase is elevated cellular (Na+), but how this is translated into a mechanism for altering the specific clearance of this enzyme from the membrane is not known. PMID- 6292953 TI - Surface and cytoplasmic domains in polarized epithelial cells. PMID- 6292954 TI - Prenatal diagnosis by restriction analysis: methodology and experience. PMID- 6292955 TI - High performance liquid chromatography (HPLC) of arachidonic acid metabolites. PMID- 6292956 TI - [New studies relating to the preservation of aqueous solutions of vancomycin, neomycin and polymyxin B]. PMID- 6292957 TI - Trimethoprim-polymyxin ophthalmic solution versus chloramphenicol ophthalmic solution in the treatment of bacterial conjunctivitis. AB - Forty patients with a diagnosis of presumptive bacterial conjunctivitis were treated with either trimethoprim-polymyxin or chloramphenicol ophthalmic solutions in a randomized, double-blind trial. No significant differences could be demonstrated between the results of 7-days' treatment with the two preparations and both solutions were found to be effective in improving the signs and symptoms of surface ocular bacterial infection. No adverse reactions were reported in either treatment group. PMID- 6292958 TI - Effects of central and peripheral pretreatment with fluoxetine in gustatory conditioning. AB - The administration of fluoxetine, a relatively specific serotonin uptake inhibitor, an hour prior to a taste-drug pairing was shown to attenuate the acquisition of taste aversions in a dose-dependent manner. Desipramine which is less effective than fluoxetine in blocking the reuptake of serotonin was also less potent in reducing the magnitude of taste aversions. Depletion of forebrain serotonin by lesions of the dorsal and median raphe nuclei or of norepinephrine by lesions of the dorsal noradrenergic bundle failed to prevent the pretreatment effect produced by either fluoxetine or desipramine. Rats with raphe lesions consistently consumed less of the taste paired with lithium than did control animals; however, this decreased intake occurred under both drug and saline pretreatment conditions, suggesting an increased sensitivity to the taste-lithium pairing rather than a diminution of the pretreatment effect. Rats with dorsal bundle lesions failed to differentiate between drug and saline pretreatment, consuming similar amounts under both conditions. These findings as well as the observation that intraventricular administration of fluoxetine did not produce a pretreatment effect suggest that forebrain serotonergic systems are not the critical site of action for the production of pretreatment effects by monoamine uptake inhibitors. Instead, the hypothesis that the peripheral effects of fluoxetine have a stimulus value that acts by way of an associative mechanism to attenuate gustatory conditioning must be considered. PMID- 6292959 TI - Anorexia and hyperphagia produced by five pharmacologic classes of hallucinogens. AB - The acute actions of five prototype hallucinogens administered SC on food consumption in 23 hr food deprived dogs were compared with the anorexic effect of d-amphetamine and the hyperphagic effect of sodium pentobarbital. Comparisons were made on the basis of dose-response relationships. Among the hallucinogens decreasing food intake, both LSD and atropine produced substantial anorexia, but the slopes of their dose-response curves were clearly different from d amphetamine. Phencyclidine and the opioid SKF 10,047 suppressed food intake also; their individual dose-effect curves were parallel to the amphetamine curve, although both were less potent. Of the hallucinogens tested, only delta-9 tetrahydrocannabinol (delta 9-THC) stimulated food consumption, and though it was less potent, it resembled pentobarbital both qualitatively and by having a parallel dose-response curve. The appetitive responses are discussed in relation to other pharmacologic actions of these hallucinogens in the dog, and consideration is given to the possible modes of action for phencyclidine- and SKF 10,047-induced anorexia. PMID- 6292960 TI - Brain benzodiazepine receptor changes in rats with isolation syndrome. PMID- 6292961 TI - [Angiotensin I converting enzyme--properties, functions and methods of analysis]. PMID- 6292962 TI - [Comparison of the antiviral activity of oxidized caffeic acid and hydrocaffeic acid against herpesvirus hominis types 1 and 2 in vitro]. PMID- 6292963 TI - [Antiviral activity of phosphonoacetic acid, phosphonopropionic acid, and trisodium phosphonoformate hexahydrate against herpesvirus hominis type 1 and type 2 in vitro]. PMID- 6292964 TI - A second generation catecholamine hypothesis. AB - The catecholamine hypothesis proposed an increase in functional brain norepinephrine in the manic phase of manic-depressive illness. This paper reviews a secondary hypothesis suggesting that dopamine may be involved in the manic process. Increases in functional dopamine may result from the release of dopamine which is amplified by a supersensitive dopamine receptor that has developed during the depressive phase prior to the onset of mania. Lithium may act, in part, by blocking the development of supersensitive dopamine receptors. Evidence compatible with this hypothesis is reviewed. PMID- 6292965 TI - Kinetics of simian virus 40 and lambda inactivation by photoaddition of psoralen derivatives. PMID- 6292966 TI - Blockage of restriction endonuclease cleavage by thymine dimers. PMID- 6292967 TI - Sex differences in biobehavioral responses to conflict in a taste aversion paradigm. AB - A conditioned aversion to a novel milk solution was produced, and animals were then reexposed to milk while nondeprived (low conflict) or following a 72-hr food and water deprivation regimen (high conflict). No sex differences occurred if animals were nondeprived throughout testing. However, if deprived during the interval between conditioning and reexposure, sex differences in both behavior and adrenocortical responses occurred: (1) Presession corticoid levels of females were higher than those of males. (2) On the first reexposure day, females showed a suppression of plasma corticosterone below presession levels, while males maintained elevated or increased corticosterone levels. (3) On the second reexposure day, when no longer deprived, males showed a marked suppression of intake compared to females, and females subsequently recovered to pretoxicosis intake levels faster than males. (4) Gonadectomy eliminated these sex differences. While ovariectomized females continued to resemble intact females in both behavioral and hormonal responses, castrated males exhibited a corticoid suppression on the first reexposure day, and subsequently recovered to pretoxicosis intake levels at the same rate as females. PMID- 6292968 TI - Replication of plasmid chimera pBR-mtB-A containing rat liver mtDNA fragment in Escherichia coli polA cells. PMID- 6292969 TI - Wide host range cloning vectors: a cosmid clone bank of an Agrobacterium Ti plasmid. PMID- 6292970 TI - Sequence rearrangements in the plasmid ColV,I-K94. PMID- 6292971 TI - A restriction map of the IncI 1 plasmid TP110. PMID- 6292972 TI - Plasmids and transposons acquired by Salmonella typhi in man. PMID- 6292973 TI - Method for the transfer of large cryptic, non-self-transmissible plasmids: ex planta transfer of the virulence plasmid of Agrobacterium rhizogenes. PMID- 6292974 TI - Nutrition and the etiology of cancer. AB - There is a growing body of scientific evidence that diet and nutrients play a causative role in the formation of cancer and that these factors may be amenable to active methods of prevention. It has been estimated that up to 35 per cent of cancers may be preventable by dietary manipulation. A theoretical basis for this are the direct-acting carcinogens ingested or produced in the diet, but these are not a major factor in the United States. Altered intake, metabolism, and excretion of cholesterol and bile acids by bacterial flora may be an important mechanism related to several commonly occurring cancers. Fiber and other constituents of vegetables may act as important protectors against the development of cancer. Dietary components related to cancer include dietary fat, fiber, fruits and vegetables, protein, alcohol, vitamins, and minerals. Practical advice for the primary care physician is given regarding two common food additives, saccharin and nitrites. PMID- 6292975 TI - [Current status of knowledge on the changes in the hypothalamo-hypophyseo-adrenal cortex system and their effects in chronic alcohol use and withdrawal]. AB - The results of investigations into the changes in the function of the hypothalamus-hypophysis-renal gland system in chronic alcoholics are collated. The review shows that the activity of the hypothalamus-hypophysis-renal gland system is below-normal in at least some alcoholics, but due to conflicting results, it is not possible to decide whether the hypofunction of the adrenal gland is a primary of a secondary effect. The interrelationship between glucocorticosteroids and catecholamines is described and its importance for the adaptability of the organism is stressed. On the basis of the review it may seem advisable to introduce corticosteroid treatment into the therapeutic schema for delerious alcoholic patients and also to apply it on patients with the alcohol withdrawal syndrome if tests show that the function of their hypothalamus hypophysis-renal gland systems is impaired. PMID- 6292976 TI - [Differentiation between Wernicke encephalopathy and cranial alcoholic polyneuropathy by measurement of brain stem conduction time (preliminary contribution)]. PMID- 6292977 TI - [Noradrenergic neurons, their interactions and the mechanism of affective disorders. Facts and hypotheses]. PMID- 6292978 TI - [The endorphin hypothesis of schizophrenia]. PMID- 6292979 TI - Lack of effect of raphe lesions on serotonin S2 receptor changes induced by amitriptyline and desmethylimipramine. AB - Spiperone binding was studied in rat cortical and striatal homogenates by direct binding assay after chronic (10 mg/kg/day i.p. for 21 days) treatment with the tricyclic antidepressants amitriptyline (AMT) and desmethylimipramine (DMI). Both AMT (a potent serotonin reuptake inhibitor) and DMI (a weak serotonin reuptake inhibitor) treatment were associated with decreased specific binding of 3H spiperone and 3H-mianserin in the cortex (markers for serotonin S2 receptors) of 31-45% of normal rats, but not the binding of 3H-spiperone in the striatum (a marker for dopamine D2 receptors). The same chronic treatment of raphe-lesioned rats with AMT or DMI produced equivalent (38-40%) decreased specific binding of these ligands in the cortex but no change in striatum. These results suggest that synaptic serotonin concentration may not be the factor responsible for the decreased number of spiperone binding sites observed after chronic antidepressant treatment. PMID- 6292980 TI - Changes in locomotor response to beta-endorphin microinfusion during and after opiate abstinence syndrome--a proposal for a model of the onset of mania. AB - Beta-Endorphin (0.3 or 0.6 nanomoles) was infused into the A10-ventral tegmental area (VTA) of male Wistar rats previously treated for 6 days with either morphine sulfate or lactose via subcutaneously implanted silastic pellets. Beta-Endorphin microinfusions occurred at 24 and 96 hours after pellets were removed. Profound changes in locomotor response to beta-endorphin were found, with morphine pretreated rats showing a spontaneous switch from hyporesponsiveness to hyperresponsiveness over 72 hours, compared to lactose-pretreated controls. These findings may reflect on current biochemical theories regarding the "switch" process in bipolar affective disease. The data can be viewed within a heuristic model of receptor changes which may underlie the transition from depression to mania. PMID- 6292981 TI - Response of plasma beta-endorphin immunoreactivity to d-amphetamine and placebo in schizophrenic patients. AB - The response of plasma beta-endorphin (ir) to infusions of randomly assigned d amphetamine (20 mg) and placebo was studied in eight schizophrenic patients. Although there was no statistically significant difference between the response to d-amphetamine and placebo, significant increases in plasma beta-endorphin (ir) levels were observed following each infusion. Although heterogeneity in beta endorphin (ir) response was observed, individual differences did not relate to clinical variables such as abnormalities on computed tomography or "process reactive" distinctions. An excessive beta-endorphin response to placebo in schizophrenia is discussed. PMID- 6292982 TI - Discriminative stimulus properties of narcotic and non-narcotic drugs in rats trained to discriminate opiate kappa-receptor agonists. AB - The purpose of this study was to evaluate the discriminative stimulus properties of some narcotic and non-narcotic drugs in rats trained to discriminate the effect of the proposed opiate kappa-receptor agonists ethylketocyclazocine and bremazocine. Male Sprague-Dawley rats were trained in a two-lever food-reinforced paradigm to discriminate between the effect of ethylketocyclazocine (0.32 mg/kg) or bremazocine (0.04 mg/kg) and saline. Both groups of trained rats showed dose dependent generalization to the effect of the proposed kappa-agonist MRZ-2033 and some animals generalized the effect of nalorphine and pentazocine. Some ethylketocyclazocine - but no bremazocine - trained rats generalized the effect of buprenorphine. The effect of dextrorphan, phencyclidine, and ketamine was generalized by some bremazocine -, but no ethylketocyclazocine-trained rats. Neither group of rats generalized the effect of etorphine, haloperidol, diazepam, or pentobarbital. These data suggest the usefulness of this procedure to evaluate the kappa-like properties of opioid drugs. PMID- 6292983 TI - Effects of GABAergic agonists and antagonists on various ethanol-induced behavioral changes. AB - The interaction between ethanol and various GABAergic drugs (muscimol, bicuculline, picrotoxin) with regard to their effects on locomotor activity, drug induced sleep, body temperature, and convulsions was studied. It was demonstrated that the GABA receptor agonist muscimol potentiated the sedative properties of ethanol, while the opposite effect, a reduction of ethanol-produced sedation, was seen upon administration of the GABA receptor blocking agent picrotoxin. Consequently, the results from the present series of experiments indicate that ethanol enhances central GABAergic activity. PMID- 6292985 TI - Neurotransmitter receptors: heterogeneity, regulation, and function. PMID- 6292986 TI - Monoamine receptor sensitivity and the mechanism of action of antidepressant treatment. PMID- 6292984 TI - Benzodiazepine antagonist Ro 15-1788: neurological and behavioral effects. AB - In neurological and behavioral studies in mice, rats, dogs and squirrel monkeys, the imidazobenzodiazepinone Ro 15-1788 acted as a potent benzodiazepine antagonist. The antagonistic activity was both preventive and curative and seen at doses at which no intrinsic effects were detected. It was highly selective in that it acted against CNS effects induced by benzodiazepines but not against those produced by other depressants, such as phenobarbitone, meprobamate, ethanol, and valproate. The onset of action was rapid even after oral administration. Depending on the animal species studied, the antagonistic effects lasted from a few hours to 1 day. The acute and subacute toxicity of Ro 15-1788 was found to be very low. Benzodiazepine-like effects were not seen. PMID- 6292987 TI - The indole hallucinogens--a new look at their mechanisms of action. PMID- 6292988 TI - Biochemical and behavioral studies of anxiety. PMID- 6292989 TI - The prevalence of cytomegalovirus antibody in women: an epidemiological study from South Wales. PMID- 6292990 TI - Effect of ionizing irradiation on the physiological activity of cyclic adenosine monophosphate on smooth muscle preparations. AB - The effect of ionizing irradiation on the physiological activity of cyclic adenosine monophosphate (cAMP) in smooth muscle preparations from frog lung was studied. cAMP, given as dibutyryl salt (dib-cAMP) inhibited the radiation induced contractions of the muscle in a manner similar to the action of theophylline. In vitro irradiation of dib-cAMP resulted in an alteration of the chemical structure of this substance, i.e., formation of monobutyryl-cAMP and further derivatives as well as a decomposition of the purine structure. There was also a loss of the relaxing activity of irradiated cAMP on the muscle tone of frog lung preparations. The physiologically measured inactivation of dib-cAMP was far more pronounced than the chemical alteration. An inhibitory effect of the reaction products is postulated. PMID- 6292991 TI - Energy deposition spectra of pi- calculated from pion nucleus interaction data. AB - Distributions of absorbed dose in linear energy transfer (LET) and in lineal energy (y) are calculated for beams of negatively charged pions in a water phantom. The calculation is based on a comprehensive set of experimental data. The production of delta-ray electrons by fast particles is taken into account semiempirically. The results are compared with experimentally obtained spectra of ionization yields. The equivalence of data derived from pion nucleus interaction and taken from microdosimetry is clearly revealed. The distribution of absorbed dose is given in a sequence of contributions from the various secondary particles, i.e., the so-called 'star' particles emitted following a nuclear capture process or the recoil nuclei from pion scatterings. This unique feature of calculated spectra will be useful for a characterization of the beam quality in view of the existing dependence of biological effects on track structure properties. PMID- 6292992 TI - [Ribosephosphate pyrophosphokinase activity in the thymus and liver of irradiated mice]. PMID- 6292993 TI - [Arteriographic diagnosis of soft tissue neoplasms of the extremities]. PMID- 6292994 TI - Pancreatic venous sampling and arteriography in localizing insulinomas and gastrinomas: procedure and results in 55 cases. AB - A comparative study of arteriography and pancreatic venous sampling (PVS) was performed in 55 patients. Twenty-seven patients with Zollinger-Ellison syndrome, 24 with insulinomas, and 4 control subjects underwent arteriography and PVS (for pancreatic hormonal radioimmunoassays) in an attempt to localize a suspected endocrine tumor. Accurate tumor localization was achieved by arteriography in 13% of the cases of gastrinoma and 29% of the cases of insulinoma. Although arteriographic signs could be described retrospectively in 62% of insulinomas, erroneous localization was relatively frequent. In contrast, localization by PVS was successful in 36 out of the 38 patients who underwent surgery; false-negative results were obtained in two patients, but in no case did PVS result in false localization. In 27% of insulinomas and 43% of gastrinomas, gross examination during surgery was negative but microscopic tumors were identified. PMID- 6292995 TI - EOE-13 in the detection of hepatosplenic lymphoma. AB - Thirty-nine patients with lymphoma were evaluated prospectively to determine the usefulness of Ethiodol-Oil-Emulsion-13 (EOE-13) in the detection of hepatosplenic lymphoma by computed tomography. The detection rate in the spleen increased from 8% (before EOE-13 infusion) to 92% (after EOE-13 infusion). In ten of 39 patients (25%) in this series, lymphomatous disease was recognized only on the postinfusion computed tomographic scan. The postinfusion EOE-13 study demonstrated additional visceral abnormalities in 38% of the patients. The potential usefulness, limitations, and toxicity of this hepatosplenic-specific imaging agent are discussed. PMID- 6292996 TI - Bone scintigraphy in plasma-cell myeloma. A prospective study of 70 patients. AB - Radiography and scintigraphy were correlated in 70 patients with recently diagnosed, untreated multiple myeloma, including 59 with and 11 without primary lytic bone lesions. A site-by-site comparison showed that scintigraphy was more sensitive than radiography in only 18% of cases, whereas radiography was more sensitive in 38% (p less than 0.001). Patients whose bone scan was as sensitive or more so than the radiograph ("hot" myeloma) had more active disease than those with the "cold" form. Remission was indicated by significant regression or disappearance of scintigraphic abnormalities in 90% of cases. The authors conclude that scintigraphy is not helpful in detecting myelomatous bone lesions, but does have prognostic value for diagnosis and chemotherapy: a positive bone scan indicates initial or residual activity. PMID- 6292998 TI - Calcium physiology in smooth muscle. PMID- 6292997 TI - [Immune response in guinea pigs to foot-and-mouth disease virus. Study of the biological activities of IgG subclasses in hyperimmunized animal sera]. PMID- 6292999 TI - Mechanism of drugs action on ion and water transport in renal tubular cells. PMID- 6293000 TI - Wilms' tumor: an approach to vena caval intrusion. AB - Wilms' tumor invades the renal vein in 12% of patients and extends into the inferior vena cava in another 6%. In a few cases, the tumor propagates into the right side of the heart. Since patients with Wilms' tumor seldom require renal arteriography, and because vena cava involvement may be silent, venous invasion may go undetected until the time of radical nephrectomy. A preoperative vena cavogram is recommended, and if a thrombus is found, a right heart angiogram is needed to determine the superior extent of thrombus. An understanding of the collateral venous circulation of the kidneys and of the physiological effects of malignant or iatrogenic obstruction of the inferior vena cava are essential to formulating the operative approach. Various operative maneuvers, intraoperative studies, and surgical aids (including cardiopulmonary bypass and hypothermia) have made forms of intracaval Wilms' tumor amenable to excision. PMID- 6293001 TI - Prostaglandin F2 alpha antagonizes thromboxane A2-induced human platelet aggregation. AB - The effects of prostaglandin F2 alpha on human blood platelet function were investigated. PGF2 alpha at 15 muM completely blocked platelet aggregation induced by 500 muM arachidonic acid or 3 muM U46619 but had no effect on aggregation induced by 7.5 muM ADP. A similar specificity of action was not obtained with either PGI2 or PGE2. Thus concentrations of PGI2 (3 nM) or PGE2 (20 muM) which inhibited U46619- induced aggregation by 100% also blocked ADP stimulated aggregation. The inhibitory properties of PGF2 alpha were not related to increases in platelet cAMP, since direct measurement of intracellular cAMP revealed that 15 muM PGF2 alpha produced no substantial change in cAMP levels. This finding was in direct contrast to results obtained using induced significant increases in platelet cAMP levels. The possibility that PGF2 alpha directly interacts at the platelet TXA2/PGH2 receptor was investigated by measuring [3H]PGF2 alpha binding to isolated platelet membranes. It was found that [3H] PGF2 alpha binding reached equilibrium within 30 min at room temperature and could be 90% displaced by addition of 1000 fold excess of unlabelled PGF2 alpha. Furthermore, when 1000 fold excess of either the TXA2/PGH2 "mimetic' U46619 or the TXA2/PGH2 antagonist displaced by 95% and 85% respectively. In contrast, the same molar excess of 6-keto-PFG1 alpha, azo analog 2, or TXB2, caused displacement of only 15%, 20% or 25% of the [3H] PHF2 alpha binding. Scatchard analysis indicated that [3H] PGF2 alpha has two binding sites; i.e., a high affinity binding site with an apparent Kd of 50 nM and a low affinity binding site with apparent Kd of 320 nM. These results suggest that the selective inhibition by PGF2 alpha of AA or U46619- induced aggregation may be mediated through interaction at the platelet TXA2/PGH2 receptor. PMID- 6293002 TI - A radioimmunoassay for leukotriene B4. AB - A radioimmunoassay for leukotriene B4 has been developed. The assay is sensitive; 5 pg LTB4 caused significant inhibition of binding of [3H]-LTB4 and 50% displacement occurred with 30 pg. The specificity of the assay has been critically examined; prostaglandins, thromboxane B2 and arachidonic acid do not exhibit detectable cross-reactions (less than 0.03%). However, some non-cyclic dihydroxy- and monohydroxy-eicosatetraenoic acids do cross-react slightly (e.g. diastereomers of 5,12-dihydroxy-6,8,10-trans-14-cis-eicosatetraenoic and 12 hydroxy-5,8,10,14-eicosatetraenoic acids cross-react 3.3% and 2.0% respectively). The assay has been used to monitor the release of LTB4 from human neutrophils in response to the divalent cation ionophore, A23187. The immunoreactive material released during these incubations was confirmed as LTB4 by reverse phase high pressure liquid chromatography following solvent extraction and silicic acid chromatography. PMID- 6293003 TI - Effect of a test meal, duodenal acidification, and tetragastrin on the plasma concentration of beta-endorphin-like immunoreactivity in man. AB - The effects of various test materials on plasma beta-endorphin-like immunoreactivity (beta-EpLI) were investigated in man using a specific radioimmunoassay developed by the authors. Plasma beta-EpLI was determined after extraction by the acid/acetone method (recovery 73 +/- 5%). The intraassay and interassay coefficients of variation were 5.0% and 7.6%, respectively. The plasma concentrations of human beta-EpLI in normal subjects were 11.6 +/- 4.0 pmol/l for men (n = 23) and 10.7 +/- 4.8 pmol/l for women (n = 27). Ingestion of a test meal (150 g of Campbell's condensed meat soup) resulted in a biphasic rise in plasma beta-EpLI from the basal level of 4.4 +/- 1.0 pmol/l to 29.2 +/- 1.9 pmol/l after 5 min and 24.8 +/- 6.7 pmol/l after 90 min. Intraduodenal infusion of 115 ml of 0.1 M HCl over 10 min increased the plasma beta-EpLI level from 8.7 +/- 0.5 pmol/l to 15.5 +/- 0.4 pmol/l at 10 min after the start of infusion, but the level rapidly returned to the initial value after the end of the infusion. Intramuscular injection of 4 micrograms/kg body weight of tetragastrin markedly stimulated gastric acid output and beta-EpLI release, but pretreatment with 10 mg of histamine H2 receptor antagonist inhibited the gastric acid output and plasma beta-EpLI release induced by tetragastrin. These results indicate that beta-EpLI release is stimulated by ingestion of meat soup, duodenal acidification and tetragastrin administration. It is suggested that gastric acid participates, at least in part, in postprandial release of beta-EpLI, probably from the gastrointestinal tract. PMID- 6293004 TI - Distribution of luteinizing hormone-releasing hormone receptors in the rat ovary. AB - The distribution of luteinizing hormone-releasing hormone (LHRH) receptors was studied in the adult rat ovary using autoradiography after injection of the stable LHRH agonist 125I-labelled [D-Ser(TBU)6,des-Gly-NH2(10)]LHRH ethylamide (Buserelin) and by radioreceptor assay using the same tracer. In intact cycling female rats, no differences in ovarian LHRH receptor levels could be observed between day diestrus I and day proestrus. Moreover, similar levels are observed in total ovarian homogenate, corpora lutea and the remaining ovarian tissue in adult animals treated with PMSG (pregnant mare's serum gonadotropins) and hCG (human chorionic gonadotropin). Radioautographic data show a comparable distribution of grains over theca interna and externa, granulosa and luteal cells. The present findings indicate the presence of LHRH receptors in both the interstitial and follicular cells throughout all stages of cellular differentiation. PMID- 6293005 TI - Secretin exerts inhibition of the 8BrcAMP-stimulated 86Rb influx into avian red blood cells. AB - Avian erythrocytes possess a Na+ and K+ cotransport system which can be inhibited by loop diuretics. The newly discovered diuretic effect of secretin in man led us to study the effect of this hormone on the cotransport system. Secretin caused a 50% inhibition of the 8BrcAMP-stimulated 86Rb influx into red blood cells from goose at a concentration of 8.5 X 10(-6) M, while furosemide and bumetanide caused a 50% inhibition at concentrations of 7 X 10(-6) M and 9 X 10(-8) M, respectively. It is suggested that the diuretic effect of secretin is mediated through an inhibitory or blocking effect on the Na+ and K+ cotransport system. PMID- 6293006 TI - [Renal angiomyolipoma. 1st case seen in the Hospital Santo Tomas]. PMID- 6293007 TI - [Tumors of the lacrimal caruncle]. PMID- 6293008 TI - [Multiple bone metastases of hepatocarcinoma without pulmonary metastases]. PMID- 6293009 TI - [Parasitization by Fasciola hepatica. Diagnostic keys]. PMID- 6293010 TI - [Physiology of the renal handling of phosphate]. PMID- 6293011 TI - [Anatomo-clinical correlations of bronchopulmonary cancer. Apropos of 72 cases]. PMID- 6293012 TI - [Periodic hemofiltration]. PMID- 6293013 TI - [Age determination in Djallonke sheep in the North of Ivory Coast by examination of their dentition]. PMID- 6293014 TI - [Isoenzymatic forms and distribution of adenylate kinase and creatine kinase in the bovine adrenal medulla]. AB - Adenylate kinase, creatine kinase as well as their substrate and product levels have been investigated in the adrenal medullary tissue. The concentration of adenine nucleotides and creatine + creatine phosphate are 12.6 +/- 0.4 and 6.9 +/ 0.4 mumol/g wet weight respectively. Adenylate kinase is mainly in the cytosol; only 4% was found in mitochondria. The cytosol enzyme presents a Km for AMP of 5 X 10(-4) M and a Ki for diadenosine pentaphosphate of 0.6 X 10(-6) M. In gel electrophoresis, only one band of adenylate kinase activity can be seen, and its mobility is different from that of the brain enzyme. Creatine kinase from adrenal medulla is mainly found in cytosol; only 3-4% was associated with mitochondria. The cytosolic enzyme is mainly the BB isozyme form. PMID- 6293015 TI - [Contribution of brain and liver to the biosynthesis of cholesterol during the postnatal development of the chicken]. AB - The high content of esterified cholesterol found in neonatal chick liver seems to be transferred from the yolk and decreases throughout the yolk sac regression, increasing at the same time the percentage of free cholesterol newly synthesized. In brain, there are no significant changes in cholesterol content during postnatal development, being predominant the free cholesterol synthesized during myelination. Changes in the hepatic HMG-CoA reductase activity are similar to those found in the acetate incorporation into nonsaponifiable lipids and are in agreement with the inhibition of cholesterogenesis during the first days after hatching and its activation with the decrease of esterified cholesterol. Chick liver PPMVA decarboxylase shows similar changes, suggesting an important role for this enzyme in the hepatic cholesterol synthesis. Only scant variations are exhibited by the MVA-activating enzymes. Brain enzymes do not change significantly throughout postnatal development. PMID- 6293016 TI - [Analysis of chromosomal and extrachromosomal functions involved in the replication of bacterial plasmids]. AB - A variety of functions codified by both the host chromosome and the plasmid genome are necessary for plasmid replication. We have found that dna B and dna F are essential for replication of both types of plasmids: R144 and Sa which have a restricted control of replication, and R6K with a relaxed control; on the other hand plasmids belonging to the first group absolutely depend on dna A and dna E gene products. Plasmid Sa, which shares with the 1st group a restricted control of replication but appears on a number of intermediate copies, shows a partial requirement to the dna A gene product. Location studies of plasmid-coded genes, involved in the replications of plasmids, were carried out in plasmid pSC102, a derivative of the resistance plasmic R6-5. All the genes required for replication are contained into a DNA segment which consists of two Pst I fragments: P-4 and P 6. The origin of replication On V is not itself sufficient for autonomous replication but needs in addition the presence of the Rep A gene. Genes responsible for copy control and incompatibility Cop/Inc are located on fragment P-6, i.e. they are physically distinct from Ori V. This indicates that the origin of replication is not responsible for, although may participate in, the incompatibility phenomenon. PMID- 6293017 TI - [Changes in hydrolytic lysosomal enzymes of liver associated with protein and protein-calorie malnutrition]. AB - The effect of a low protein (1%) diet (protein-deficient diet) and a low protein calorie diet (restricted diet) on the activity of the hydrolytic enzymes acid ribonuclease, acid deoxiribonuclease, acid and alkaline phosphatases and beta glucuronidase has been studied in the liver of Wistar rts. Experimentation was carried out over 30 days and then comparisons were made against well-nourished (10% protein, controls), and one another. Body weight of deficient animals decreased in deficient animals, especially in protein-deficient rats. Liver weight also dropped significantly in malnourished rats. In terms of organ weight relative to body weight, there was a clear increase of protein-deficient rats, compared with controls. Enzyme activities expressed per total organ fell significantly in deficient rats compared with controls, but alkaline phosphatase activity increased. A large increase in hydrolytic activity expressed per mg of protein in beta-glucuronidase and alkaline phosphatase values was registered in protein-deficient and restricted rats. Also, acid ribonuclease activity increased in deficient-protein animals, but it fell in protein-calorie deficient animals compared with controls. However, acid deoxiribunuclease and acid phosphatase activities were not modified. We therefore concluded that protein-deficient and restricted diets increase catabolism in liver through a modulation of lysosomal hydrolase activities. PMID- 6293018 TI - [Biotechnology. Genetic engineering. Chemico-pharmaceutical area]. AB - We have tried to bring up to date the information on in vitro Genetic Manipulations and the Recombinant DNA considering its relations with new technology and possible applications towards the fine chemicals development and specially drug area. Taking into account the scientific development which takes place in the most advanced industrial societies, those countries which are nowadays developing an adequate technology level must face the scientific challenge among them, we can find two which appear as specially important: the Genetic Engineering knowledge and the Molecular Biology developments. PMID- 6293019 TI - [Glycosidases of various mollusks: general properties, kinetic studies and action on natural substrates]. AB - The beta-N-acetylhexosaminidase, beta-glucuronidase, alpha-galactosidase, beta galactosidase and alpha-L-fucosidase activities, in six different species of molluscs, have been studied. The optimum pH was acid in all cases, in agreement with the lysosomal origin of these enzymes. They generally show several pI in their isoelectrofocusing profiles. Kinetic studies with enzymes having several activities in one protein, i.e. beta-N-acetylhexosaminidase and beta galactosidase, have been carried out with mixed substrates in order to determine the occurrence of several active sites. The action of these enzymes on glycosidic rests containing natural substrates has been studied by enzymatic hydrolysis. PMID- 6293020 TI - [Metabolism of the organic phosphate regulators of oxygenation in cells of the erythrocyte series in birds and mammals]. AB - In mammals, bisphosphoglycerate-synthase activity, whose assay methods are previously discussed, increases gradually along erythropoiesis, leading to a consequent enhancement of 2,3-bisphosphoglycerate formation. Avian erythrocytes, on the other hand, contain inositol-pentaphosphate as major organic phosphate starting from egg eclosion, which substitutes embrionary ATP and 2,3-BPG in the regulatory function. The IHP of phytase and the disappearance of 2,3-BPG synthesis, also inhibitor of enzyme activity, should be considered responsible for IPP accumulation. PMID- 6293021 TI - [After the lunatic asylum]. PMID- 6293022 TI - Efficacy of orally-administered chelating agents for nickel carbonyl toxicity in rats. AB - The oral efficacy of several chelating drugs, disulfiram, sodium diethyldithiocarbamate (dithiocarb), and D-penicillamine, was studied in relation to their ability to prevent lethality due to acute inhalation exposure to nickel carbonyl. Dithiocarb and, to a lesser degree, D-penicillamine were found to be especially effective as therapeutic agents. Since dithiocarb is chemically unstable (although it is an active metabolite of disulfiram), its practical usefulness outside the laboratory is limited. Therefore, further investigation was conducted to elucidate the pharmacokinetic differences between dithiocarb and disulfiram in order to explain their relative efficacies. Kinetic analysis indicated that oral dithiocarb provided moderately high but prolonged plasma levels of active chelating agent, while disulfiram resulted in a very high but transient level. These results suggested that small, repeated oral doses of disulfiram would be just as effective in nickel carbonyl poisoning as a single large dithiocarb dose. This was not borne out in subsequent experiments. Investigation into the tissue distribution of inhaled 63Ni-labeled nickel carbonyl 24 hours after exposure showed that all three chelating agents significantly reduced the amounts of nickel in heart and lung, and D penicillamine further reduced the amounts in blood and kidney. Only disulfiram increased on average the nickel retained in brain tissue, possibly accounting for its limited efficacy. Our results suggest caution in the use of oral disulfiram in human nickel carbonyl intoxication. PMID- 6293023 TI - Studies on rat liver mitochondria. II. Formation of lipid peroxides in mitochondria preserved at 0-4 degrees C. AB - Levels of lipid peroxides were monitored in freshly isolated rat liver mitochondria, aged mitochondria, and preserved mitochondria by reaction with TBA (2-thiobarbituric acid) and spectrophotometric absorption at 233 nm. In aged mitochondria, the levels of lipid peroxides increased gradually during 120 hr, while in preserved mitochondria they increased rapidly up to 24-48 hr. Under such conditions, the respiratory control coefficients and the levels of total Pi were about 40% those of fresh mitochondria, and the decrease of the levels of lipid peroxides followed the decrease of the respiratory control coefficients and levels of total Pi. These results suggest that stimulation of the formation of lipid peroxides occurs when the vital activity of mitochondria is preserved. PMID- 6293025 TI - Enzymatic impairment induced by biological aldehydes in intact rat liver cells. AB - The addition of lipid peroxidation end-products, 4-hydroxypentenal (HPE), 4 hydroxynonenal (HNE) or hexanal (HEX) to the incubation medium of rat hepatocytes caused a significant decrease of cytochrome P-450 content and inactivation of total cell glucose-6-phosphatase. In particular, the two hydroxyalkenals exerted their inhibitory action at micromolar concentration. These results on intact cells further support a possible damaging effect of aldehydes in pathological conditions in which a stimulation of lipid peroxidation occurs. PMID- 6293024 TI - Promotion of benign hyperplastic lesions by calcium, magnesium and cAMP, and inhibition of tumor progression by magnesium in hamster cheek pouch. AB - Ca, Mg (0.1 or 0.2M) and ionophore A23187 (10(-5) - 10(-4)M) promoted benign hyperplastic lesions (BHLs) to a highly variable degree when applied topically in hamster cheek pouch after initiation with 7, 12-dimethylbenz(a)anthracene (DMBA). The changes in the promoting effects of Ca and Mg between experiments were not parallel. Mg but not Ca inhibited retinyl acetate-induced progression of BHLs to advanced tumors. Cyclic adenosine 3',5'-monophosphate (cAMP) (10(-5) - 10(-3)M) promoted BHLs. The results are discussed in terms of possible mechanisms involved. PMID- 6293026 TI - Production of murine cytotoxic T lymphocytes by bluetongue virus following various immunisation procedures. AB - The induction of bluetongue virus specific cytotoxic T lymphocytes (CTLs) in C3H mice by various live and inactivated bluetongue virus preparations was studied. Live virus preparations were shown to induce good levels of CTLs; however, inactivation of virus preparations either by beta propriolactone or glutaraldehyde induced only a low level response. The use of Freund's adjuvants and double immunisation procedures failed to improve the response of the inactivated preparations. These findings are discussed in relationship to protection from bluetongue disease with various bluetongue virus vaccines. PMID- 6293027 TI - Resolution of the multiple inert gas method for estimating VA/Q maldistribution. AB - During steady-state infusion of a mixture of dissolved inert gases, their elimination by the lung depends on the distribution of ventilation/perfusion (VA/Q) ratios. Thus, certain features of the VA/Q distribution can be inferred from inert gas measurement. Because of the: (1) complexity of the lung, and (2) experimental errors, the ability of such a technique to describe the shape and position of the VA/Q distribution accurately is limited. In this report we present an analysis of the resolution of the method for 9 representative sets of inert gas data, taking account of both of the above factors. These 9 sets span the range of commonly observed data, both in health and in diseases such as asthma, interstitial fibrosis, chronic obstructive lung disease and respiratory distress syndromes. Both error-free and error-containing data are studied and by linear programming methods, bounds are placed on maximum and minimum possible perfusion in several regions of the VA/Q spectrum. Modality is also studied by linear programming. The results show that the resolving power of the method depends greatly on the specific case under study. When groups of units are separated in VA/Q by a decade, this can be determined with considerable confidence. Shunt and low VA/Q areas can generally be well resolved, but when distributions are very broad, resolution is limited. PMID- 6293028 TI - [Indications for exofacial parotidectomy in the treatment of mixed tumours]. PMID- 6293029 TI - The surgery of mixed salivary tumours. PMID- 6293030 TI - [Cystic adenoid carcinoma of the salivary glands]. PMID- 6293031 TI - [Prostaglandins and related substances]. PMID- 6293032 TI - [Modern morphological study of the peripheral nerves]. PMID- 6293033 TI - Persistent infection of humans with hepatitis B virus: mechanisms and consequences. AB - Although most infections of humans with hepatitis B virus are self-limited, up to 10% of individuals experiencing primary infection remain persistently infected. These individuals are usually asymptomatic but may suffer episodic or progressive liver injury that can result in cirrhosis and liver failure. In addition, chronic infection with hepatitis B virus is, in certain settings, associated with the late development of primary hepatocellular carcinoma. This review summarizes the biological characteristics of persistent viral infection, with particular emphasis on host and viral factors that affect the outcome of primary infection. PMID- 6293034 TI - Epstein-Barr virus: a human pathogen inducing lymphoproliferation in vivo and in vitro. AB - Epstein-Barr virus (EBV) is a pathogen that is associated with several diseases in humans. It causes most heterophile-positive cases of infectious mononucleosis. The virus is associated with another lymphoproliferative disease, African Burkitt's lymphoma, and with a malignancy of the nasopharynx, nasopharyngeal carcinoma. EBV appears to be a causative agent for some fatal lymphoproliferations in congenitally immunodeficient people. One proliferating cell type in infectious mononucleosis is an EBV-infected B lymphoblast. The tumor cell in African Burkitt's lymphoma is also an EBV-infected B lymphoblast. In vitro, EBV induces and maintains blast transformation of human B lymphocytes. All EBV-transformed cells, whether infected in vivo or in vitro, share many (but not all) virus-associated characteristics. Studies can now be performed in vitro that permit analysis of the various EBV-transformed cells and the host's immune responses to them. PMID- 6293035 TI - The Paul-Bunnel test revisited. PMID- 6293036 TI - A successful eradication campaign. Global eradication of smallpox. AB - Smallpox was the first important disease to be eradicated; it was the success of the Smallpox Eradication Programme that inspired this conference. Several biological reasons favored the eradication of smallpox, the most important of which were probably that recurrent infectivity did not occur, that there was no animal reservoir, and that an effective stable vaccine was available. The importance of smallpox as a disease that travelers might import into countries free of smallpox provided a powerful stimulus for its global eradication. This paper highlights some of the problems associated with the eradication of smallpox in two countries where eradication was difficult, India and Ethiopia, and the measures adopted to overcome the problems. The paper also stresses the importance of the development of methods for the certification of smallpox eradication from countries, from regions, and finally from the whole world. It is noted that close links between field work and research were important throughout the eradication campaign. PMID- 6293037 TI - [Obesity, energy expenditure and thyroid hormones]. PMID- 6293038 TI - [Determination of dietary fiber in foods by the enzymatic method]. PMID- 6293039 TI - [Effect of sunlight on the concentrations of 3,4-benzopyrene in the atmosphere of Madrid]. PMID- 6293040 TI - Herpes simplex virus specific secretory IgA in lacrimal fluid during herpes keratitis. AB - The correct diagnosis of herpes keratitis is very important, as the clinical manifestation can be similar to that caused by immunologic defects. The presence of herpes simplex virus (HSV) specified IgG in lacrimal fluid is not of diagnostic value in cases where the eyes are severely inflamed, because the inflammation leads to impairment of the blood vessels and allows a passive diffusion of IgG from serum into the lacrimal fluid. In the present study we have developed a method for the detection of locally produced HSV-specific antibodies of the secretory IgA class present in lacrimal fluid from patients with active herpes keratitis. PMID- 6293041 TI - A hypothesis on the development of acute myocardial infarction in Greenlanders. AB - Non-emigrated Greenlanders have a low incidence of acute myocardial infarction (AMI), when compared with age- and sex adjusted death rates for ischemic heart disease in western countries. We find that Greenlanders have plasma lipid levels corresponding to favourable risk factor levels for AMI. This can be attributed to their diet, rich in n-3 polyunsaturated fat. This diet further supplies eicosapentaenoic acid which influence platelet vessel wall function in an antithrombotic direction. A high level of plasma-antithrombin-III, raising the anticoagulant activity of the blood, in combination with a genetically high activation threshold for the complement system may further contribute to the resistancy against thrombo-embolic disorders. Bleeding tendency, and susceptibility to infection disorders may be the possible draw-backs. Our data are framed into a hypothesis combining the indications of genetic predispositions and the evidence of exogenous protective factors, inflicting a coherent enhancement of nonsusceptibility to vascular ischemic catastrophies. PMID- 6293042 TI - Antacids and HCl. PMID- 6293043 TI - Antacid therapy--changes in mineral metabolism. AB - Antacid ingestion may lead to side-effects related to their chemical composition. Aluminum hydroxide may cause the phosphate depletion syndrome even during short term administration of high doses in patients at high risk, such as alcoholics. Long-term intake may lead to bone demineralization and to osteomalacia. Fluoride complexing in the gut and prevention of fluoride absorption may be an additional factor. The clinical relevance of aluminum absorption in patients with normal renal function is not clear. In contrast, in patients with renal failure, aluminum hydroxide ingestion may contribute to an increasing hyperaluminemia. Hyperaluminemia and tissue deposition of aluminum in these patients may contribute to the dialysis-associated encephalopathy. Magnesium hydroxide causes an alkalinization of the urine due to magnesium absorption and urinary excretion. Thus, in renal insufficiency, a life-threatening hypermagnesemia may develop if magnesium-aluminum-containing antacids are prescribed. The milk-alkali syndrome, rarely observed nowadays, may be caused by calcium carbonate- and sodium bicarbonate-containing antacids. Hypercalciuria and alkaluria predispose to nephrolithiasis. The possibility that these disturbances in mineral metabolism will develop in patients with normal renal function is unlikely unless there is an abuse of these "over the counter" antacids. PMID- 6293045 TI - Non-A, non-B hepatitis after open-heart surgery in Sweden. AB - In recent years evidence has emerged that most post-transfusion hepatitis is caused by one or more previously unknown agents named non-A, non-B. A prospective investigation was made of 74 patients who underwent open-heart surgery. Only volunteer blood was used for transfusions. Transfusion-associated hepatitis appeared in 15 (20%) of the patients 4-12 weeks after the operation. In no case was the hepatitis found to be caused by hepatitis B, A or Epstein-Barr virus. One patient had a cytomegalovirus infection; the other 14 cases (19%) were classified by definition as non-A, non-B hepatitis. Although most of the patients were asymptomatic and all were anicteric, the course of the hepatitis was protracted in many cases. Thus, 6/12 observed patients still had pathologic transferase values more than a year after the onset of hepatitis. Liver biopsy was performed in 3 cases and showed histologic signs of chronic active hepatitis in all of them. PMID- 6293044 TI - Transferrin receptors on mitogen-stimulated human thymus-derived lymphocytes. AB - The appearance of transferrin receptors on mitogen-stimulated human thymus derived (T) lymphocytes was studied. When indirect immunofluorescence with immunoadsorbent-purified antitransferrin antibodies was used, approximately 10% of resting T cells were stained. This proportion increased to 50-80% of the cells 3-4 days after stimulation with the mitogenic lectins concanavalin A (Con A) and leucoagglutinin (La) from Phaseolus vulgaris. Almost all blast cells (greater than or equal to 90%) were positive. Cell binding experiments with 125I-labelled transferrin indicated the presence of 1-5 x 10(5) transferrin receptor molecules/cell with high avidity for transferrin (K = 2 - 12 x 10(8) l/mol). Analysis by sodium dodecyl sulphate polyacrylamide gel electrophoresis and autoradiography of cell lysates containing 125I-labelled T-cell surface components revealed two surface peptides (90 kdaltons and 80 kdaltons, reducing conditions), which selectively bound to insolubilized antitransferrin antibodies. The 90-kdalton peptide also bound to insolubilized transferrin. The 80-kdalton peptide is most probably transferrin and the 90-kdalton peptide the transferrin receptor. Unreduced transferrin receptor had a molecular weight of 180 kdalton. It is probably a glycoprotein, since it reacted with wheat germ agglutinin, La, and probably also Con A. The properties of the lymphocyte transferrin receptor are similar to those described for transferrin receptors on various in-vitro grown transformed cells. This speaks in favour of a common receptor present on all proliferating human cells. PMID- 6293046 TI - Acute encephalitis of viral origin. AB - 75 patients with acute encephalitis of confirmed or suggestive viral or virus like etiology showed a predominance of herpes simplex virus (HSV) as an etiological agent (13.3-32.0%) in all age groups without seasonal variation and increasing in number during years. Mumps, occurring mainly in female patients, was next in order and thereafter coxsackievirus B, varicella-zoster, and adenoviruses, followed by multiple coincident infections, and a large number of occasional agents. HSV seems to have displaced tick-borne encephalitis virus as the major cause of acute encephalitis in Finland. Pregnancy appeared to increase susceptibility to HSV encephalitis. The outcome was associated with the etiological agent in the first hand, HSV and nonviral microbes appeared unfavourable, but also with the clinical symptomatology, particularly altered consciousness, mental symptoms and focal neurological signs. The need for exact and rapid diagnosis in all cases of acute encephalitis is underlined in view of the possibility of specific therapy. PMID- 6293047 TI - Epidemiological studies of congenital cytomegalovirus infection. PMID- 6293048 TI - [Effect of ACTH administration before and after nitrogen oxide exposure on lung histamine and serum corticosterone concentrations in rats]. PMID- 6293049 TI - Cytomegalovirus infection and kidney graft survival. AB - Among 123 cadaveric renal allograft recipients transplanted in the period 1971 79, there were 18 with no evidence of past or present cytomegalovirus infection, 34 with primary infection and 71 with reactivated infection. One-year actuarial graft survival was 68%, 32% and 54%, respectively. The reasons for the better graft prognosis in the group without CMV infection were less rejection and fewer infections. PMID- 6293051 TI - [Effect of ACTH on plasma cortisol levels and hematological parameters in the horse]. PMID- 6293050 TI - [Phagocytes and phagocytosis 100 years after Metchnikoff. A current picture of the neutrophil leukocyte]. AB - The observation of the amoebocytes of primitive organisms led ELIAS METSCHNIKOFF in 1882 to the idea that blood phagocytes--neutrophilic leukocytes in particular- could constitute an anti-microbial defense system. This was the beginning of the phagocyte theory which METSCHNIKOFF developed over many years and which in essence is still valid. The author sets out to provide an updated view of the neutrophil. Circulating neutrophils are end-cells. They develop in the bone marrow by a relatively long maturation process during which the characteristic azurophil and specific granules are formed. The granules are stored organelles. The azurophil granules contain microbicidal enzymes, i.e. myeloperoxidase and lysozyme, together with a large number of acid hydrolases and neutral proteases. The specific granules contain lysozyme, a collagenase, lactoferrin and transcobalamines. By subcellular fractionation a third kind of storage organelle has recently been found which is characterized by its gelatinase content. Circulating neutrophils are activated on microbial invasion--first in the blood, by chemotactic factors formed at the site of infection, and subsequently by the microbes themselves which are phagocytosed by the immigrating neutrophils. Chemotactic factors lead to directed migration and induce the secretion of enzymes which presumably facilitate this process. Phagocytosis results in the mobilization of neutrophil products in large quantities. The contact between the cell and the microorganism activates in the neutrophil membrane an oxidase which produces superoxide, and a phospholipase which releases arachidonic acid. The latter is then oxidized by cyclooxygenase and lipoxygenase. There is also massive liberation of enzymes from all three storage compartments. The production of superoxide is the essential process for the killing of a large variety of microorganisms. PMID- 6293052 TI - New method for detecting cellular transforming genes. AB - Tumor induction in athymic nude mice can be used to detect dominant transforming genes in cellular DNA. Mouse NIH 3T3 cells freshly transfected with either cloned Moloney sarcoma proviral DNA or cellular DNA's derived from virally transformed cells induced tumors when injected into athymic nu/nu mice. Tumors were also induced by cells transfected with DNA from two tumor-derived and one chemically transformed human cell lines. The mouse tumors induced by human cell line DNA's contained human DNA sequences, and DNA derived from these tumors was capable of inducing both tumors and foci on subsequent transfection. Tumor induction in nude mice represents a useful new method for the detection and selection of cells transformed by cellular oncogenes. PMID- 6293053 TI - In vivo identification of the transforming gene product of simian sarcoma virus. AB - Simian sarcoma virus (SSV) deletion mutants were constructed from a molecular clone containing the entire infectious provirus. Transfection analysis of these mutants localized the SSV transforming gene to a small region of the viral genome encompassing its cell-derived sequence (v-sis). Antiserum to a peptide synthesized on the basis of the predicted amino acid sequence of the SSV transforming gene detected a 28,000-dalton protein that was specifically expressed in SSV transformed cells and that corresponded in size to that predicted from the v-sis coding sequence. The v-sis gene product designated p28sis was not a phosphoprotein, nor did it possess detectable protein kinase activity. These findings distinguish p28sis from a number of other retroviral onc proteins. PMID- 6293054 TI - Crypts are the site of intestinal fluid and electrolyte secretion. AB - The site of adenosine 3',5'-monophosphate-mediated fluid and electrolyte secretion across mammalian large intestine was found to be the crypts of Lieberkuhn by means of two techniques. First, the formation of fluid droplets was visualized on the oil-covered mucosal surface directly over crypt duct openings when secretion was stimulated. Second, microelectrode impalement of individual surface and crypt cells revealed that only crypts cells produced a pattern of secretagogue induced alterations in membrane potential and resistance that was characteristic of secretory epithelia. PMID- 6293055 TI - Fragment spanning the SV40 replication origin is the only DNA sequence required in cis for viral excision. AB - A 311-base pair fragment containing the SV40 origin of replication was linked to the chicken thymidine kinase gene on a recombinant plasmid. This molecule was transfected into human 143 thymidine kinase-deficient (TK-) cells, and colonies positive for thymidine kinase were selected. When cell lines derived from these colonies were fused to permissive simian cells that produce SV40 T antigen, the recombinant plasmid excised itself from the human cellular genome and replicated with a high copy number per cell. These results show that this segment of the viral genome is the only sequence required in cis to mediate SV40 excision and replication upon fusion to permissive cells. In addition, we have shown that excised plasmids apparently identical to the input DNA can be efficiently rescued in Escherichia coli. SV40 excision and replication may therefore be useful for the recovery of cloned genes from eukaryotic cells. PMID- 6293056 TI - Eukaryotic transcriptional regulation and chromatin-associated protein phosphorylation by cyclic AMP. AB - Cyclic adenosine monophosphate (AMP) analogs or agents that increase intracellular cyclic AMP rapidly stimulate transcription of the prolactin gene in a line of cultured rat pituitary cells. This effect is correlated with the phosphorylation of a chromatin-associated basic protein designated BPR. These data are consistent with the postulate that increased intracellular cyclic AMP concentrations induce rapid transcriptional effects on specific genes in eukaryotes, mediated by direct or indirect phosphorylation of a specific chromatin-associated protein or proteins. PMID- 6293057 TI - Chromosomal assignment of the endogenous proto-oncogene C-abl. AB - Abelson murine leukemia virus (A-MuLV) is a replication-defective retrovirus that transforms lymphocytes of the B-cell lineage. This virus is a recombinant between the parental Moloney murine leukemia virus and a cellular gene termed C-abl. By analysis of a series of mouse x Chinese hamster hybrid celllines containing various mouse chromosomes, we have mapped the C-abl gene to mouse chromosome 2. PMID- 6293058 TI - Yeast mating pheromone activates mammalian gonadotrophs: evolutionary conservation of a reproductive hormone? AB - alpha-Factor, a tridecapeptide mating pheromone of yeast (Saccharomyces cerevisiae), has extensive sequence homology with the hypothalamic decapeptide gonadotropin-releasing hormone (GnRH). Both synthetic and natural preparations of alpha-mating factor were found to bind specifically to rat pituitary GnRH receptors and to stimulate the release of luteinizing hormone from cultured gonadotrophs. The ability of the yeast pheromone to reproduce the biological actions of GnRH in the mammalian pituitary gland indicates that the structural and functional properties of GnRH-related peptides may have been highly conserved during evolution. PMID- 6293059 TI - Benzodiazepine receptor-mediated experimental "anxiety" in primates. AB - The ethyl ester of beta-carboline-3-carboxylic acid has a high affinity for benzodiazepine receptors in the brain. In the rhesus monkey this substance produces an acute behavioral syndrome characterized by dramatic elevations in heart rate, blood pressure, plasma cortisol, and catecholamines. The effects are blocked by benzodiazepines and the specific benzodiazepine receptor antagonist Ro 15-1788. The benzodiazepine receptor may consist of several subsites or functional domains that independently recognize agonist, antagonists, or "active" antagonists such as beta-carboline-3-carboxylic acid ethyl ester. These results suggest that the benzodiazepine receptor is involved in both the affective and physiological manifestations of anxiety, and that the administration of beta carboxylic acid ethyl ester to monkeys may provide a reliable and reproducible animal model of human anxiety. PMID- 6293060 TI - [Immunopharmacology of cancer: a new appraisal and clinical needs]. PMID- 6293061 TI - [Progress in the detection and changes in the treatment of prostatic cancer]. AB - Treatment of cancer of the prostate has been deeply transformed by the advent of D-TRP6-LH-RH. This hormonal agonist does not induce hyperestrogenemia with it's cardiovascular and glandular complications (painful gynecomastia). D-TRP6-LH-RH gives a 74% remission rate. Chemotherapy of cancer of the prostate is not toxic (vincristine, ifosphamide, 5-fluorouracile and pepleomycine); as it gives a 24% remission rate it may be that chemotherapy induces remission in tumors without androgen receptors while D-TRP6-LH-RH is only effective against tumors in which these receptors are present. This implies that assay of androgen receptors now determines the therapeutic indications for cancer of the prostate. Estrogen therapy, even in low doses, does not prolong survival of patients as compared to untreated controls, because of iatrogenic vascular complications. It will therefore be abandoned. As for local therapy, indications should be limited to failures of D-TRP6-LH-RH and chemotherapy. PMID- 6293062 TI - [Phase II study of 3 new nitrosoureas, 1 American (chlorozotocin) and 2 French (RFCNU and RPCNU)]. AB - The results of three phase II clinical trials with three new analogues belonging to the family of nitrosoureas are reported. One of the studied agents, chlorozotocine (CZT) is American, while the other two, RFCNU and RPCNU, are French. All three drugs have a sugar radical. CZT proved effective mainly in a few cases of leukemia and in one case of blastic transformation of chronic myelocytic leukemia. RFCNU was shown to be effective in 8% of digestive tumors, in 1 out of 7 pancreatic cancers and in 3 out of 10 hepatic and pulmonary metastases from an undiscovered primary adenocarcinoma. As for RPCNU, it's action resembles that of RFCNU: tumor regression lasting for over three years was obtained in a patient with hepatic metastases from a digestive carcinoma; in another patient a regression rate exceeding 50% was seen in pulmonary metastases from a rectal tumor. One of the significant results of our study is the apparent tissular specificity of responses according to the agent given: CZT is more often efficient on the lymphatic localizations of the studied tumors (4/5 responses); RFCNU and RPCNU often proved active on hepatic metastases (17% responses). Digestive tolerance was excellent with CZT and RFCNU and not quite as satisfactory for RPCNU. As predicted by our experimental study, platelet toxicity is both less common and less severe with RFCNU than with CZT and RPCNU. PMID- 6293063 TI - [Treatment of rectocolic and gastric adenocarcinomas with 5-fluorouracil associated with high doses of folinic acid. Results of an experimental study]. AB - We report the results of a therapeutic trial in patients with rectocolic and gastric metastatic adenocarcinomas. This trial is based on experimental evidence that an excess of reduced intracellular folic acid increases the cytotoxicity of fluoropyrimidines. The treatment consists of 5-fluorouracile (5-FU) (370 to 400 mg/m2/24 h) and folinic acid in high doses (200 mg/m2: 24 h) given simultaneously for 5 consecutive days; the interval between courses is 21 days. Thirty patients with measurable rectocolic adenocarcinomas were evaluated. They were divided into two groups: 16 patients had had no previous chemotherapy and 14 had not responded to chemotherapy with 5-FU alone or associated with other cytostatic drugs. Response rates were 56% in the first group and 21% in the second. Five patients with measurable gastric adenocarcinomas were also evaluated; none had received previous chemotherapy. A partial response was recorded in three of these patients. Toxicity of the therapeutic regimen was acceptable. Stomatitis was the most common toxic side-effect. In patients with severe adverse side-effects recurrence was efficiently prevented by decreasing the daily dose of 5-FU to 30 mg/m2 during subsequent courses. We conclude that in the tumors studied folinic acid in high doses can improve the antitumoral effect of 5-FU and induce a response to this agent in some rectocolic tumors which were previously resistant. PMID- 6293064 TI - [Currently recommended use of the Hemoccult for the detection of rectocolic tumors]. AB - Because colorectal carcinoma is both very frequent and extremely serious, diagnosis and treatment of this cancer are a major Public Health issue in France. The recognition of the filiation between polyps and cancer has led to considering prevention of colorectal cancer through the detection of polyps. The usefulness of the hemoccult test is discussed. Reported results warrant the rejection of this method for the individual detection of colorectal tumors because of poor sensitivity. Conversely, there are grounds for advocating the use of the hemoccult test for mass screening on certain conditions; the most important of these is that a negative test must not be considered as sufficient to eliminate the diagnosis. If screening plans with the hemoccult test are implemented, they will have to be coordinated with an important health education program to promote individual prevention using endoscopy, particularly in high risk and symptomatic subjects. PMID- 6293065 TI - [Pleural mesothelioma: 3 developmental forms]. AB - In the medical literature, there have been fewer reports on benign pleural mesotheliomas than on their malignant counterparts. We have reviewed the clinical, radiological and pathological features of mesotheliomas seen at the Hotel-Dieu in Montreal over the past ten years. This experience leads us to postulate that there are three types of mesothelioma. The malignant form is often typical. Pedunculated lesions most often prove to be asymptomatic and benign. However, we have identified an intermediate category where the definitive nature of the mesothelioma could only be established after long term follow-up of the biological behaviour of the tumor. PMID- 6293066 TI - [Rectal stenosis caused by cancer of the prostate: difficulties of diagnosis]. AB - There exist rare cases in which a prostatic carcinoma invades the perirectal space producing an extrinsic rectal stenosis. Three cases are discussed. Differential diagnosis with a rectal tumor is difficult clinically, but deep rectal biopsies will provide the correct diagnosis in many cases. Estrogen therapy is recommended, but prognosis is reserved. PMID- 6293067 TI - [Perfusion and ventilation pulmonary scintigraphy in the diagnosis of the operability of primary bronchial cancers]. AB - Ventilation scintigraphy using Munsch's technique with Xenon 133 combined with perfusion scintigraphy using 99m Tc albumin particles in microspheres are of considerable interest in the preoperative evaluation of bronchogenic carcinoma. Basing themselves on 95 cases, the authors discuss the manner in which the tumor affects ventilation and perfusion. In the 70 cases of matching ventilatory and perfusion scans, scintigraphy alone is not sufficient to demonstrate the exact perfusion obstruction mechanism. On the other hand, if there is a V/Q mismatch (21 cases) and if the perfusion defect is more extensive than the ventilation defect or the radiological opaque area, this normally implies a mediastinal extension of the tumor, and tends to make any carcinologically valid ablation highly risky. Comparison of the perfusion and ventilation scans with the radiological data and spirometric values showed that 9 patients presented with abnormal pulmonary radiographs outside the tumor areas. Six of these cases involved tuberculous sequelae. Forty-four of the ninety five patients (46.3%) showed normal ventilation and perfusion scans outside the tumor areas. Of the 91 patients undergoing spirometry, 39 suffered from ventilatory obstruction and pathological ventilation and perfusion scans and 18 presented with scintigraphic anomalies outside the tumor are a but did not suffer from ventilatory obstruction. PMID- 6293068 TI - [Isotopic scintigraphy in the diagnosis of the spread of bronchial cancer]. AB - Ventilation of the lungs using Xenon 133 and perfusion scans using 99m Tc albumin particles in microspheres were performed on 455 patients with bronchial cancers and 55 patients with pulmonary metastases. Ventilation-perfusion mismatch and functional defects of the lung, which are never found in nodular-shaped pulmonary metastases, vary in frequency according to the anatomo-pathological type of the tumor and constitute a limiting factor for ablation. PMID- 6293069 TI - [Chronic lymphoid leukemia with erythroblastopenia and primary liver tumor]. AB - Erythroblastopenia occurred in the course of chronic B cell lymphocytic leukemia. The failure of chlorambucil therapy prompted the decision of thymic irradiation. This was effective on the lymphoid proliferation but did not modify the erythroblastopenia. Progressive hepatomegaly led to the diagnosis of hepatoma. PMID- 6293070 TI - [Primary hepatic carcinoma disclosed by Budd-Chiari syndrome. Apropos of a case. Review of the literature]. AB - Hepatic carcinoma revealed by Budd Chiari syndrome in an eighty-four-year-old female patient is reported. With reference to this observation and to the twenty two other cases previously reported in the medical literature, the specific pathological and clinical features of Budd Chiari syndrome caused by primary carcinoma of the liver are reviewed. Histological features of the liver tumour are discussed. PMID- 6293071 TI - [Baso- and spinocellular Pinkus fibroepithelial tumor]. AB - A seventy-eight-year-old man presented with a moist proliferating tumor of the back which arose six months earlier amidst numerous verrucae seborrheic. The tumor's size was 20 X 28 mm. Two different aspects were found upon histologic examination. One was a Pinkus fibroepithelial tumor with foci of basal cell proliferation, stromal changes with fibrosis, and lymphocytic infiltration. Adjacent to this tumor was a squamous-cell carcinoma. Foci of squamous-cell carcinoma were also found within the Pinkus epithelial tumor. PMID- 6293072 TI - [Effect of tiapride on headache and various other types of pain]. AB - The effects of tiapride were studied in 180 patients, including 165 with cephalalgia originating in various causes and 15 with other types of pain. 110 of the 165 patients with cephalalgia completed the study; results were good or excellent in 78 (71%), with no differences related the the cause of the headache. 13 of the 15 patients with other types of pain completed the study, with good or excellent results in 10. Tiapride was given in a daily dose of three tablets a day in outpatients, and two daily intramuscular injections in hospitalized patients. Tolerance was excellent in 109 of the 123 patients (88%). Recorded side effects were drowsiness in 4 patients, asthenia in 3, ebrious manifestations in 3, amenorrhea-galactorrhea in 2 and constipation in 2. PMID- 6293075 TI - [Puncture biopsy in the diagnosis of papillary epitheliomas of the thyroid]. AB - In this study of the cytological appearances of papillary adenocarcinoma of the thyroid after fine needle aspiration biopsy, the authors have brought together 60 examinations of thyroid nodules and 10 indicant ganglial metastases. They isolate three significant aspects. type I (25 cases) is notable for the dense cellular desquamation in papillary clusters with angular contours, numerous calcospherites and, above all, the presence of intranucleaur pseudo-inclusion bodies (PI) which are clearly visible after coloring with May Grunwald stain. Type II (17 cases) was mainly observed in cystic lesions (8 cases) and metastatic ganglia (6 cases). It shows thick clusters of necrotic cells, few PI, barely visible against a background preparation of lysated hemates, polynuclear cells and macrophages. Type III (19 cases) consists of layers of cells poor in cytonuclear anomalies. The presence of PI is both a prerequisite and sufficent arounds for a diagnosis of papillary carcinoma of the thyroid, which will always be confirmed by subsequent histopathological examinations. The absence of PI leads to a false negative result. Better knowledge of cytological aspects specific to papillary tumors of the thyroid improves the quality of the diagnosis, not in terms of the percentage of carcinomas identified, which rises from 75.6 to 77% at a second reading, but in the specification of the papillary type, which rises from 38.5 to 64.2%. In the absence of PI and cytonuclear anomalies, non-identification of the carcinoma (22.8%) results from uninterpretable smears (5.7%) and false negatives (17.1%). There were no false positives. Cytological examination after fine needle aspiration biopsy should be included in the preoperative examinations of all thyroid nodules, as well as in the exploration of cervical ganglia likely to host an indicant or delayed metastasis. PMID- 6293074 TI - [Syndrome of inappropriate antidiuretic hormone secretion. A report of two cases with hyponatremia and hypouricemia]. AB - Two cases of Schwartz-Bartter syndrome are reported. Both were due to malignant anaplasic tumours of the APUD type with multiple abnormal endocrine secretion, and both were accompanied with hypouricaemia of uncertain significance. The authors believe that the association of hypernatraemia with hypouricaemia should alert clinicians to the possibility of a syndrome of inappropriate antidiuretic hormone secretion (SIADH) of malignant origin. PMID- 6293073 TI - [Management of advanced breast cancer in post-menopausal women. A comparative trial of hormonal therapy, chemotherapy, and a combination of both]. AB - A randomized trial was done in 98 post-menopausal women with breast cancer. Hormonal receptors had not been assayed in any patient. Patients were given hormonal therapy with tamoxifen and drostanolone propionate (n = 34), or chemotherapy with the CMF protocol (n = 30), or a combination of both (n = 34). The results are in favour of hormonal therapy alone, which gave the best immediate objective responses, the least adverse side-effects, and possibly improved survival rates. PMID- 6293076 TI - [Biliary hamartoma. Reflexions on a new personal case]. AB - In this study of a personal case of biliary hamartoma showing complete aplasia of the hepatic parenchyma as well as a cystic ectasy of the corresponding bile ducts, particular emphasis has been laid on the clinical, paraclinical and anatomo-pathological aspects of these dysgeneses. There seems to be an analogy between the radiographic, anatomical and anatomo-pathological studies made in this case and those that could have been made in pulmonary sequestration. As regards the pathogenesis of these complaints, the authors therefore put forward the same hypothesis for biliary hamartoma as for pulmonary sequestration and highlight the advantages of pre-operative angiography. PMID- 6293077 TI - [Study of the effectiveness of tiapride on psychic disorders in alcoholics]. AB - The effectiveness of tiapride on psychic disorders was studied in 30 chronic alcoholics by assessing four parameters: sleep disturbances, mood disorders, conduct disturbances, and tremor. In a daily dosage of 600 to 900 mg tiapride proved particularly effective on insomnia, anxiety, passivity and tremor, without adverse side-effects. This drug seems useful for controlling psychic disorders in alcoholics and for motivating acceptance of a detoxification program. PMID- 6293078 TI - [A giant-cell tumor of the mandibular condyle]. AB - The authors present a case of a giant cell tumor of the mandibular condyle, which is a very rare condition. Clinical history and symptoms suggested a disease of the ear. There was a destruction of the cortex of the condyle an invasion of the joint cavity, and an erosion of the adjacent temporal bone. PMID- 6293079 TI - [Necrotizing enterocolitis. A study of forty-six cases seen in a maternity hospital]. AB - Forty-six cases of necrotizing enterocolitis were seen at the Baudelocque maternity hospital from august 1978 to october 1980. In eighteen cases, clinical signs associated with roentgenograms showing pneumatosis led to unequivocal diagnosis. A surgical procedure was done in nine of these patients, during the acute stage in four and later on in five. Diagnosis was strongly suspected in twenty-eight cases. Clinical features were less serious and included abdominal distension, bloody stools, and emesis. There were no roentgenological signs. None of these infants underwent surgery. Overall mortality rate was 7% (3/46). In comparison to previously published studies, our series shows lower perinatal risk factors, higher mean birthweight, and lower prematurity rate (14%). No evidence was found to support the responsibility of a specific bacterial agent. Corona virus was found in the stools of five out of eleven cases studied during the second epidemic wave in october 1979, suggesting a possible viral etiology. Several preventive steps have been taken. Breast-feeding has been encouraged. Each mother-infant pair has been isolated instead of grouping infants in night nurseries. Staff members have been given specific information on the means of preventing contamination. PMID- 6293080 TI - [Pathologic hyperprolactinemia. I. Positive diagnosis and etiology]. AB - Hyperprolactinemia is common and mainly encountered in premenopausal women. The most prevalent causes are drugs (estrogens, neuroleptics), hypothalamic and pituitary disorders (functional abnormalities, intrasellar adenomas, suprasellar lesions) and hypothyroidism. Although the typical picture is the amenorrhea galactorrhea syndrome, hyperprolactinemia may be revealed by many other features, including obesity, hirsutism and sterility. When plain roentgenograms show a normal sella, neither dynamic tests nor polytommography can lead to unequivocal diagnosis of a tumor. Computerized tomography scanning is now the most reliable investigation in patients with hyperprolactinemia resulting from an obvious or suspected tumor. PMID- 6293081 TI - [Serum profile of inflammatory reaction proteins in arteriopathies of the lower limbs, stage II]. AB - The serum content of proteins involved in the inflammatory process was investigated, partly through kinetic immunodiffusion, in 85 men subdivided as follows: 45 men (aged 57 +/- 5 years) with intermittent claudication due to arteriosclerosis obliterans; 20 controls matched for sex and age; 20 sex-matched controls whose mean age was 25 +/- 3 years. Patients with obliterating arteriosclerosis had signs of both subacute and chronic inflammation. Compared to controls, they had a higher sedimentation rate, a lower percentage of serum albumine, higher percentages and absolute serum contents os alpha 1, alpha 2 and beta globulins, and higher serum concentrations of fibrin, orosomucoid, C3 complement fraction and IgA. In addition, the modifications of these proteins were closely correlated. These results are consistent with previous reports concerning patients with coronary heart disease. They point out the relationship between inflammation and atherosclerosis. PMID- 6293082 TI - [Rheumatic manifestations in the presence of antibodies to Yersinia pseudotuberculosis. Apropos of 4 cases]. AB - Rheumatic manifestations due to Yersinia pseudotuberculosis are apparently rare. We report four personal observations, including one of a patient with Reiter disease and significant titres of antibodies to Yersinia pseudotuberculosis. In all four patients a non-specific syndrome, with fever and digestive symptoms, preceded the onset of joint disease. Both peripheral and axial joints were affected. One patient had HLA B27; the brother of another patient had spondyloarthropathy with HLA B27. Stool cultures were negative but they were all done at least three months after the acute episode. These rheumatic manifestations have many features in common with those described in Yersinia enterocolitica infections. Studies of larger series could improve our knowledge of the part played by the patients' constitution and widen the scope of reactive arthritis. PMID- 6293084 TI - [Acute renal insufficiency in spontaneous septicemia of renal origin. Analysis of a series of 17 patients]. AB - Seventeen recent cases of spontaneous renal septicemia with acute renal failure are reviewed, confirming the need for a diagnostic and therapeutic strategy to restore the fluid balance, if necessary with the aid of an artificial kidney, and the urgency of treating local infection and the pathogenic agents involved. The case review also suggest that the high mortality rate of this infection, estimated by some observers at 50 p. cent, can be greatly reduced by early radical urological treatment. PMID- 6293083 TI - [Thymolipomas simulating cardiomegaly]. AB - With reference to three personal observations, the authors discuss the problems raised by the diagnosis of thymolipomas, which are benign asymptomatic tumors discovered on a plain chest film and simulating cardiomegaly. On careful analysis of a high voltage plain chest film, the pulmonary vasculature can be detected through the cardiomediastinal opacity. Total body scan is also useful for establishing diagnosis before surgery. PMID- 6293085 TI - [Indirect evaluation of effort angina. Automatic and traditional analysis of the ST segment, the amplitude of the R wave and thallium scintiscanning]. AB - Forty-six patients presenting typical or non-typical angina pectoris were all submitted to analysis of effort angina, traditional or automatic, and to analysis of the amplitude of the R wave and to an effort myocardial scan using thallium 201 (taken as reference substance) during maximal muscle exercise. The following conclusions may be drawn from this study. 1) Autoanalysis of the ECG improves considerably the sensitivity without changing the specificity of the method (92 and 82 p. cent for 79 and 82 p. cent). Analysis of the variations in amplitude of the R wave gave no further information. It should be reserved for patients whose effort ECG was difficult to interpret: e.g. left or right bundle branch block, digitalis effect, very abnormal resting ECG, or average age female population. PMID- 6293086 TI - [Multiple paragangliomas of the neck localized in the thyroid region. Papillary thyroid cancer associated with parathyroid adenoma]. AB - A 58 years-old woman has had a goiter for many years. She was treated by neck irradiation for "lymphadenopathy" 20 years-ago. She is now operated on for bilateral carotid body paragangliomas and a similar tumor of the thyroid gland near the midline. She also presents a papillary carcinoma of the left thyroid lobe with lymph node metastasis and a chief-cell parathyroid adenoma. The diagnosis of thyroid paraganglioma is discussed. Then the authors raise the hypothesis of radiation-induced papillary carcinoma. They try to set a link between thyroid tumors and parathyroid adenoma. PMID- 6293087 TI - [Prophylaxis of infectious complications in maxillo-facial surgery. Value of cefazolin. (A review of 400 cases)]. AB - The value of short-term prophylaxis with antibiotics in maxillo-facial surgery and plastic facial surgery is studied. 200 patients were included in the study and compared to 200 controls who were given the usual systematic antibiotic therapy, with a different antibiotic, for more than 6 days. 400 case-reports were thus retrospectively analyzed. The results show that when the surgical procedure lasts for less than three hours, short-term prophylaxis with antibiotics is more effective than the usual systematic antibiotic therapy given for more than 6 days. PMID- 6293088 TI - [Multicentric granular cell tumor of the tracheobronchial tree]. AB - The authors describe a case of multicentric endotracheobronchial myoblastoma with one endotracheal localization and two bronchial localizations. The patient underwent resection of the inferior lobe of the left lung. The tracheal tumor was not removed. Two years after surgery the patient is doing well. Several biopsy specimens have been removed from the tracheal tumor which has remained unchanged. Cases such as ours are very uncommon. Therapeutic decisions may be difficult. PMID- 6293089 TI - [Urologic manifestations disclosing a tumor of the spinal cord. Apropos of a case, ependymoma of the terminal cone]. AB - A twenty-two-year-old man was hospitalized for acute renal failure resulting from long-standing ureterohydronephrosis. An ependymoma of the terminal spinal cord was removed. Physiopathology of micturition and the features which allow differential diagnosis are recalled. Previously reported cases are reviewed. PMID- 6293090 TI - [Consumption coagulopathy complicating Mediterranean kala-azar in an adult]. PMID- 6293091 TI - [Prediction of aminoglycoside nephrotoxicity]. AB - Mechanism of aminoglycoside nephrotoxicity is yet unclear. Reduction of 20 p. cent of glomerular filtration initial value is observed in 10 p. cent of cases. Aminoglycoside nephrotoxicity appears to be related to high doses, to duration of treatment and also to individual factors such as age and preexisting renal impairment. No mean of predicting aminoglycoside nephrotoxicity without any error is available. Meanwhile, accurate measurement of glomerular filtration and of beta 2-microglobulin urinary elimination seems clinically more usefull for detection of impaired renal function than uneasy measurement of urinary enzyme excretion and than monitoring the aminoglycosides serum concentrations. PMID- 6293092 TI - [Diaphragmatic paralysis and eventration]. AB - Paralysis of the diaphragm normally heals in a matter of weeks, provided that it does not result from permanent damage to the phrenic nerve. The most frequent cause is traumatic, a consequence of birth or a postoperative complication, especially after cardiovascular surgery. The vital prognosis may involve respiratory insufficiency and consequent ventilation problems. But the situation has greatly changed as a result of the technical progress made in respiratory reanimation. When the necessary time is available, the indications of surgical plication are becoming rarer. Diaphragmatic eventration, on the other hand, involves rarefaction or the complete disappearance of the muscle fibres of the diaphragm. Over two thirds of such cases require surgical treatment. The eventration may result from an acquired affection leading to permanent destruction of the phrenic nerve. But it may also be associated with malformations, and in that event the prognosis is definitely severe. PMID- 6293093 TI - [The factor VIII complex: hemophilia A and von Willebrand disease]. AB - The factor VIII complex is a macromolecule with two distinct components. One is the coagulation factor VIII. The other, known as the Willebrand factor, is a polymer which probably acts as a carrier for serum factor VIII. Hereditary disorders can affect either of these two components. Hemophilia A is a coagulation disorders due to decreased factor VIII coagulant activity. Increase of partial activated thromboplastin time parallels disease severity. Hematomas and hemarthrosis in large joints are the main clinical features. In von Willebrand disease, mucocutaneous bleeding is the main symptom. Diagnosis is established by demonstrating disorders of primary hemostasis: prolonged bleeding time and decreased ristocetin-induced platelet aggregation. Two forms of von Willebrand disease have been described. In the quantitative form, decreased synthesis of von Willebrand factor is often responsible for severe clinical manifestations. The qualitative form probably results from defective polymerization of von Willebrand factor subunits. In both these forms, deficient primary hemostasis is a consequence of decreased platelet adhesion to the vascular wall. Clinical and biological features of hemophilia A and von Willebrand disease, as well as their management, are discussed. PMID- 6293094 TI - The evolutionary role of prolactin in mammalian osmoregulation: effects on fetoplacental hydromineral transport. PMID- 6293095 TI - Pituitary prolactin, lactational performance and puerperal infertility. PMID- 6293096 TI - Etiology of childhood infectious mononucleosis syndrome in Singapore. PMID- 6293097 TI - Postirradiation osteogenic sarcoma with unilateral metastatic spread within the field of irradiation. Case report and review of the literature. AB - Osteogenic sarcoma is a rare but well documented complication of radiation therapy. To date, only 28 cases of radiation induced osteogenic sarcoma, following therapy for breast carcinoma, have been reported. Only one case of postirradiation sarcoma arising in the sternum has occurred and this followed irradiation to the neck. The subject of this article is therefore only the first reported case of osteosarcoma of the sternum, secondary to irradiation for breast carcinoma. A further unusual and unreported feature is the development of unilateral metastatic disease in the field of irradiation. Possible causes for this occurrence are discussed and the literature regarding postirradiation sarcoma is reviewed. PMID- 6293098 TI - [A few neurologic problems]. PMID- 6293099 TI - Regulation of contractile proteins by reversible phosphorylation of myosin and myosin kinase. AB - In vitro experiments support the ideal that the actin-activated MgATPase activity of smooth muscle myosin and myosin from nonmuscle cells is regulated by the phosphorylation of the 20,000 dalton light chain of myosin. Experiments with intact smooth muscles support this mechanism but also raise the possibility that tension may be maintained in the presence of partial dephosphorylation (12). The possibility that smooth muscle contraction may also be modulated by additional regulatory systems (13,29) is to be expected based on experience with other types of muscle. The enzyme myosin light chain kinase catalyzes the phosphorylation of the 20,000 dalton light chain of myosin. This enzyme requires Ca2+-calmodulin for activity. The activity of myosin kinases that have been isolated from avian smooth muscle cells (8) or human platelets (16) can be decreased by phosphorylation. This phosphorylation is catalyzed by cAMP-dependent protein kinase and decreases myosin kinase activity by interfering with the binding of Ca2+-calmodulin. A number of different phosphatases have been purified from smooth muscle (22). These phosphatases play an important role in determining the state of phosphorylation of myosin and myosin kinase. Two areas of particular interest at present are the regulation of phosphatase activity and the physiological significance of myosin kinase phosphorylation. PMID- 6293100 TI - Catch muscle. PMID- 6293101 TI - Obesity reconsidered. PMID- 6293102 TI - Computed tomography in the diagnosis of congenital cytomegalic inclusion disease. AB - We present two cases of congenital CID, one in a 4-month-old male infant and the other in a 6-week-old male infant. The diagnoses were confirmed by means of computed tomography, cultures, and serologic methods. The relatively noninvasive character of CT makes it an ideal diagnostic method in the neonate. With the improved visualization afforded by CT, specificity on radiographic grounds alone may become possible. PMID- 6293103 TI - Management of radiation ulcers. AB - Despite more efficient and safer technics of radiation therapy, the problem of radiation-induced injury to the skin and soft tissue persists. The problem of adequate coverage of these painful, ischemic, and fibrotic ulcers remains challenging. Split-thickness skin grafts are seldom sufficient coverage, as the graft almost always has areas that do not take. Although these areas may eventually heal by epithelialization, the result is never ideal. Most often flap coverage is required, but elevation of local flaps is jeopardized because the tissue surrounding the ulcer crater frequently has been sufficiently compromised to cause loss of at least part of the flap. In the past, this necessitated use of pedicled flaps, tubed and transposed from a distance. With the development of axial-pattern musculocutaneous and muscle flaps, as well as microvascular free flaps, the difficulty in dealing with these ulcers has been decreased. Surgeons can now recommend earlier use of adequate debridement, many times of the entire irradiated area, and immediate coverage with a well vascularized axial-pattern musculocutaneous flap or revascularized free flap. PMID- 6293104 TI - [The obzidan test in assessing bronchial patency]. PMID- 6293105 TI - Secondary drowning in the Cape Peninsula. AB - Secondary drowning is a respiratory distress syndrome which develops after a latent period following apparently successful resuscitation from near-drowning. Eighteen cases of near-drowning have been reviewed and one case history is presented in detail to illustrate the importance of anticipating the development of this syndrome. Successful management of these patients involves ventilatory support and careful monitoring of the neurological and cardiovascular status. Because many doctors are unaware of the syndrome and because it has a good prognosis if recognized and treated early, a review of the clinical features and treatment is warranted. PMID- 6293106 TI - Unusual serologic response to two patients to an early antigen of herpes simplex virus type 2. AB - Two patients infected with herpes simplex virus type 2 (HSV-2) who were free of recurrences over a 13-14-month period had an unusually high titer (greater than 1:48) of complement-fixing antibody to the early antigen (AG-4) of HSV-2. Their response was also unusual in that it was an IgG complement-fixing antibody rather than IgM, which normally is found in patients with HSV-2 infection. The antibody was also present 13-14 months after the episode of HSV-2 disease, whereas in other patients, the response had declined by six months. PMID- 6293107 TI - The hydroxyl-hydrogen ion concentration ratio during hypothermia. AB - Fundamental physicochemical characteristics of the acid-base related constituents of extracellular and intracellular fluid spaces of vertebrates in relation to changes in temperature have been reviewed. Emphasis has been placed upon the dissociation constant of water, the solubility constant of CO2, the dissociation constant of histidine imidazole, the hydroxyl-hydrogen ion ratio, the protein charge state and the alpha-imidazole regulation concept. Because pN and pKIm change in parallel when temperature varies, the OH/H ratio and the alpha imidazole value for any sample of blood or plasma held anaerobically in vitro are invariant with changing temperature, since a constant CO2 content is maintained. Thus, when blood or plasma cools, pH increases and PCO2 decreases, but relative alkalinity and the protein charge state remain constant. These responses are solely the consequence of physical constants, that is, equilibrium constants and gas solubility, changing with temperature. In vivo, the set of PCO2 is established in each poikilothermic species by its normal ventilatory pattern designed to maintain constant CO2 content. Regulation in vivo in poikilotherms consists of adjustments of ventilation per unit metabolism (VA/VCO2) appropriate to every temperature. When the ventilatory and renal mechanisms of human beings are suppressed by anesthesia and hypothermia, their extracellular and intracellular responses mimic those of poikilotherms. Clinical management of hypothermia in humans requires ventilatory control using oxygen-augmented room air without added CO2 monitored by pH measurements of arterial blood warmed anaerobically to 37 degrees C. Finally, the need for new techniques to measure intracellular pH as temperature is lowered and some areas for further investigation are suggested. PMID- 6293108 TI - Angiographic detection of unsuspected carcinoma of the thyroid gland. AB - Angiography is seldom used to detect carcinoma of the thyroid. We have recently encountered four patients with hypervascular lesions of the neck demonstrated by angiography for evaluation of an unrelated medical problem. In each, carcinoma of the thyroid was detected and total thyroidectomy performed. Because of the possibility of carcinoma of the thyroid in patients with hypervascular lesions of the neck, we recommend exploration of the neck, regardless of symptoms, physical findings or scintiscan results. PMID- 6293109 TI - Immunopathology of polyoma-induced tumors. PMID- 6293111 TI - Role of the major histocompatibility complex in resistance to viral leukemia; its effect on the preleukemic stage of leukemogenesis. PMID- 6293114 TI - Investigation of the effects of phospholipase C on human platelets: evidence that aggregation induced by phospholipase C is independent of prostaglandin generation, released ADP and is modulated by cyclic AMP. AB - Effects and the mechanism of action of phospholipase C (PLC), from Clostridium perfringens, on washed human platelets were examined to better understand the role of PLC in platelet function. PLC caused aggregation and secretion of [14C] 5HT, without concomitant loss of cytoplasmic, LDH, in a concentration dependent manner. P-nitrophenylphosphorylcholine, a substrate for PLC, blocked these responses in a concentration dependent manner. In other experiments hirudin, alpha-1-antitrypsin and soybean trypsin inhibitor did not inhibit PLC-induced activation of human platelets. PLC-induced aggregation and [14C]-5HT secretion was not inhibited by aspirin, a known inhibitor of prostaglandin biosynthesis. PLC-induced aggregation was selectively inhibited by analogs of 7,8 dihydroxybenzazepine and 7,8-methylenedioxybenzazepine in a concentration dependent manner. These two agents had no effect on arachidonic acid-induced aggregation. PLC-induced aggregation was not inhibited by apyrase, an enzyme which hydrolyzes ADP. In other experiments, PLC-treated platelets did not exhibit any platelet activating factor-like activity. Prostaglandin E1 and trifluoperazine showed concentration dependent inhibitor effects on PLC-mediated aggregation and secretion of [14C]-5HT. These findings indicate that: a) PLC is capable of inducing aggregation and specific secretion of [14C]-5HT without causing lysis of platelets; b) mechanism of PLC-induced activation of platelets is independent of prostaglandin generation or action, released ADP, and PAF; and c) cyclic AMP plays a modulatory role in PLC-mediated secretion and aggregation of human platelets. PMID- 6293112 TI - Regulation of the immune response to antigens on the malignant cell surface. PMID- 6293113 TI - Modulation of antitumor immunity--immunobiologic approaches. PMID- 6293115 TI - Obstruction of the inferior vena cava in the hepatic portion and hepatocellular carcinoma. AB - High incidence of hepatocellular carcinoma in patients with obstruction of the inferior vena cava in the hepatic portion (Budd-Chiari syndrome) was previously pointed out by us from the review of Japanese literature of autopsied cases. This was confirmed by the follow-up study of 16 patients with obstruction of the inferior vena cava in the hepatic portion hospitalized in 1958 to 1974. Follow-up information was available on 13 of the 16 patients. Deaths due to hepatocellular carcinoma occurred in 6 patients (46%), and those due to other causes in 4 patients (31%). Three patients were alive. Hepatocellular carcinoma occurred most frequently in the patients who were found to have obstruction of the inferior vena cava at less than 44 years of age and were followed up for more than 10 years. PMID- 6293116 TI - Significance of prednisolone administration for hepatic mitochondrial function of the rat with biliary obstruction. AB - Mitochondrial respiratory function of the liver is disturbed in biliary obstruction, especially in that caused by tumours in the hepatobiliary system. This study aimed to clarify whether a glucocorticoid, prednisolone succinate, is effective to improve the reduced mitochondrial function of the rat liver in obstructive jaundice. Five doses of 5 mg/kg or 25 mg/kg prednisolone succinate were administered at 5 consecutive days to the rats after 1, 3 or 6 weeks of biliary obstruction and to the rats without obstruction, and the hepatic mitochondrial function and contents of cytochromes of these rats were investigated. With 25 mg/kg prednisolone, hepatic mitochondrial function was improved in rats with biliary obstruction of 3- or 6-weeks duration as compared with the prednisolone-untreated group. Compensatory increase or recuperation of the once decreased cytochrome A(+a3) and/or turnover number of phosphorylation was also observed in 25 mg/kg group. Hepatic mitochondrial function of obstruction-free animals was rather impaired by treatment with 25 mg/kg prednisolone. In 5 mg/kg group, above mentioned effects were scarcely observed. It was concluded that the hepatic mitochondrial function in animals with prolonged biliary obstruction, as same as in animals with short-term obstruction, can be improved by the administration of adequately large amounts of prednisolone. PMID- 6293117 TI - Postischemic liver damage in rats: effect of some therapeutic interventions on survival rate. AB - The aim of this paper is to elucidate the cause of death after 90 min of normothermic partial (2/3) ischemia of the liver and to examine the effects of glucagon, somatostatin, insulin, prednisolone and oral administration of polymyxin B (PB). The animals 24 hr after partial ischemia for 90 min were divided into two groups; namely, animals with normal appearance and those with moribund state. There were no significant differences in the plasma level of S GOT, S-GPT, amino acids, NH3 or insulin, or in morphometrically estimated volume ratio of necrotic hepatocytes between the two groups of rats. The blood glucose level, however, was significantly decreased (31 +/- 28 mg/100 ml, n = 6) in the moribund rats with a higher incidence of positive Limulus gelation tests as compared with the rats with normal appearance (149 +/- 19, n = 5). The 1-day and 1-week survival rates of the animals were 42/62 (69%) and 32/61 (53%), respectively. A glucagon injection (1.5 mg/kg, after ischemia) was effective to elevate the 1-day survival rate (14/14), but failed to increase the 1-week survival rate (11/14). On the other hand, a somatostatin injection (100 micrograms/kg, after ischemia) or PB treatment (15 mg/kg/day x 5-9, before ischemia) succeeded to increase the 1-week survival rate (20/22 p less than 0.01 and 17/17 p less than 0.01, respectively), although no significant amelioration in transaminase levels or volume ratio of necrosis was demonstrated. It could be seen that a moribund state after partial ischemia was accompanied by severe hypoglycemic shock, and that the injection of somatostatin after ischemia or the annihilation of gram-negative bacteria by means of oral administration of polymyxin B before ischemia prevented the occurrence of the hypoglycemic shock. PMID- 6293110 TI - Natural and induced immunity to mouse mammary tumors and the mammary tumor virus (MuMTV). PMID- 6293118 TI - Tumorigenesis by a ferroactinolite mineral. AB - In lifetime exposure of male Fischer-344 rats to ferroactinolite fibers and to UICC amosite asbestos fibers by means of intratracheal and intrapleural treatments, oncogenesis was greater in the lung for the ferroactinolite and in the pleura for the amosite. The lack of correlation between the effects of the two methods of exposure suggests that in this instance intrapleural inoculation was not a good predictor of pulmonary response on the basis of mass dose. Another feature of the ferroactinolite was that pleural tumors resulted from intratracheal instillations. Conversely, lung tumors or tumorlike lesions were also induced by intrapleural inoculations of ferroactinolite. These facts suggest a greater in vivo transport for the ferroactinolite than for the amosite. Since there are far fewer mineral fibers per mass unit in the ferroactinolite the tumor yield per unit of mineral fibers was strikingly greater by both routes of administration for ferroactinolite than for amosite. PMID- 6293119 TI - Interpretation of the carcinogenicity of amosite asbestos and ferroactinolite on the basis of retained fiber dose and characteristics in vivo. AB - Rats were exposed to amosite asbestos and ferroactinolite fibers by intrapleural inoculation and intratracheal instillation. The ferroactinolite sample was found to be more carcinogenic in both exposures than the amosite sample on the basis of total fiber dose or fiber dose expressed for any size category of hypothetical greatest carcinogenic potency. Quantitative transmission electron microscope analysis of low-temperature ashed whole lung samples collected at different times following intratracheal instillation of fibers demonstrated that concentrations and sizes of fibers retained in rat lungs were greatly influenced by the relative ability of each mineral to undergo longitudinal splitting as a consequence of dissolution in vivo. Ferroactinolite fibers rapidly split to produce many thin fibers so that the number of ferroactinolite fibers retained in the lung 2 years after intratracheal instillation was four times greater than the number of fibers originally instilled. The number of short, thin ferroactinolite fibers retained (10-fold more than amosite) after in vivo splitting best explains the greater lung carcinogenicity of ferroactinolite compared to amosite. PMID- 6293120 TI - Pleural plaques in asbestosis: effect of Candida albicans. AB - Effect of chrysotile dust alone or together with Candida albicans administered intratracheally in guinea pigs was studied in the genesis of pleural plaques over a period of 12 months. A significant increase of mucopolysaccharides, phosphorus, calcium and -SH content was detected in pleural fluid of animals treated with chrysotile and Candida albicans together than in those treated with chrysotile or Candida albicans alone. The results suggest that an infection of Candida albicans accentuates the effect of chrysotile by altering the biochemical parameters preceding to the formation of pleural plaques. PMID- 6293121 TI - Influence of lead and nickel on passive cutaneous anaphylaxis in the rat. AB - Male rats, IOPS OFA strains, were injected s.c. with doses of lead acetate (0.5 to 30 mg Pb2+/kg/day) and nickel chloride (0.05 to 5 mg Ni2+/kg/day), for 3 days prior to being challenged for immediate hypersensitivity induced by ovalbumin/antiovalbumin system. The vascular permeability was statistically increased in animals of each group (P less than 0.001, Student's t-test). These results may indicate an action of lead and nickel salts on mast cells and their membranes, but also an action on the vasodilation induced by vasoactive amines. PMID- 6293122 TI - Transvaginal absorption and disposition of nonoxynol-9 in gravid rats. AB - The placental transfer, maternal tissue distribution and elimination of [14C] nonoxynol-9 (25 mg/kg) were studied during the first 6 h in 15-day pregnant rats treated intravaginally. Blood concentrations of 14C reached a maximum of 2.0 +/- 0.06 micrograms/ml in 1 h, declining slowly in 3 h, and remaining steady thereafter. At 6 h, the highest levels of 14C were recorded in the maternal urinary bladder, followed by those in the liver, kidney, adrenal and thyroid. The maternal brain showed lowest uptake of 14C among all tissues examined. After 6 h, the 14C concentrations in the uterus or placenta were similar to that of maternal plasma, whereas the mean concentrations of 14C both in the amniotic fluid and fetus were approx. one-third of that of the maternal plasma. About 44% of the administered dose was absorbed from the vagina, and nearly 17% of the per vaginum applied radioactivity was recovered in the urine and feces over 6 h. The results show that nonoxynol-9 (N-9) is rapidly absorbed into the blood stream from the vagina of gravid rat and that the fetal uptake of 14C is much lower than that of the placenta. PMID- 6293123 TI - [Therapeutic patient care in malignant tumors localized in the oral cavity]. PMID- 6293124 TI - [Viral warts of the oral mucosa in children]. PMID- 6293125 TI - Plasma estrone-sulfate: assessment of reduced estrogen production during treatment of metastatic breast carcinoma. AB - Highly sensitive and specific estrogen assays are required to monitor the hormonal effects of surgical adrenalectomy or pharmacologic estrogen suppression in postmenopausal women with breast carcinoma. Because the levels of plasma estrone-sulfate are 10-fold higher than its unconjugated counterpart, we developed a radioimmunoassay for estrone-sulfate to quantitate the minimal estrogen concentrations expected under conditions of endocrine gland ablation. After establishing normal ranges, we compared plasma estrone- sulfate levels and urinary conjugated estrone basally and after surgical adrenalectomy or aminoglutethimide (estrogen suppression) therapy in 23 postmenopausal women with breast carcinoma. In response to either therapy, the plasma levels of estrone sulfate fell by 63.5-79.2% (p less than .01) and conjugated urinary estrone by 85 94.5% (p less than .01) in all study days over a 12-week period. Correlation analyses yielded r values of 0.77-0.94 between conjugated plasma and urinary estrone concentrations in the surgical adrenalectomy and aminoglutethimide treated groups, respectively. No significant differences in estrone-sulfate levels were observed when comparing spontaneously menopausal and surgically castrate patients. PMID- 6293126 TI - Abrogation of proliferation and generation of cytotoxic T cells in human mixed lymphocyte culture reactions by modification of the cell surface with mitogenic oxidizing agents. AB - Multiple lectins with specificity for cell surface glyco-proteins inhibit cellular and humoral immune responses and induce transplantation tolerance. Because cell surface glycoproteins play a significant role in various immune events involving cell to cell interactions and because the mixed lymphocyte culture (MLC) reaction is a prototype of immune phenomenon involving cell to cell interactions as well as an in vitro analogue of graft-destructive immune events, the effect of modification of the cell surface with oxidizing mitogens was investigated. Treatment of responder or stimulator cells with neuraminidase and galactose oxidase (NAGO) or with sodium periodate (IO-4) resulted in marked suppression of alloantigen-induced proliferation and in vitro generation of primary cytotoxic T cells (CTLs) in human MLCs. A prominent coupling of mitogen induced proliferation to abrogation of MLC was consistently observed with modification of stimulator or responder cell surface with either NAGO or IO-4. The possibility that destruction of receptor sites and/or stimulatory units was responsible for the suppression of MLCs was excluded by restoring both proliferation and generation of primary CTLs by reduction of mitogen-oxidized cell surfaces with sodium borohydride. The ability of polyclonal activators to inhibit antigen-specific responses might be useful in abrogating unfavorable alloimmune responses. PMID- 6293127 TI - Murine cytomegalovirus infection and cyclosporin A. PMID- 6293128 TI - Melanotic progonoma of the maxilla (report of a successfully treated case and a brief review of the literature). PMID- 6293129 TI - [Urinary infection in the hospital environment]. PMID- 6293130 TI - [Radio-scintigraphic correlation in the diagnosis of bone metastases of cancer of the nasopharynx]. PMID- 6293132 TI - [Melanotic progonoma in infants: apropos of 2 cases]. PMID- 6293131 TI - [Sensitivity of 300 strains of gram-negative bacteria to cefotaxime]. PMID- 6293133 TI - Ultrastructure of congenital cystic adenomatoid malformation of the lung. AB - Four cases of congenital cystic adenomatoid malformation of the lung are presented with ultrastructural analysis. In some of the cases the ultrastructure is similar to that described in normal fetal airways during the early embryonic phase of lung development. Varying degrees of differentiation are noted, confirming what has previously been observed ultrastructurally in this lesion. One new finding is the presence of tonofilaments in one of our cases, which may represent a metaplastic change. Although the exact nature of the insult is unknown, there may be a developmental defect affecting epithelial differentiation as well as the interaction between the developing pulmonary mesenchyme and distal respiratory units. PMID- 6293134 TI - Nuclear and cytoplasmic inclusions in disseminated human cytomegalovirus infection. AB - A case of disseminated human cytomegalovirus infection is described in a full term female who expired 3 1/2 h after birth. Cytomegalic inclusions, both intranuclear and intracytoplasmic, were observed mainly in the kidneys, liver, lungs, and anterior pituitary but were not seen in the bone marrow and spleen. Whereas the nuclear inclusions consisted of an amorphous filamentous meshwork with a variable number of pleomorphic capsids, the cytoplasmic inclusions were composed of membrane-bound aggregates of mature virions with dense cores and multilayered envelopes. The complex envelopment process of cytomegalovirus appears to involve successive coats derived from the nuclear membrane and from the endoplasmic reticulum or cytoplasmic vesicles. PMID- 6293135 TI - Malignant schwannomas presenting as malignant fibrous histiocytomas. AB - Two soft tissue neoplasms considered to represent malignant fibrous histiocytomas by light microscopy showed typical findings indicative of Schwann cell origin when examined by electron microscopy. These findings included the presence of cells exhibiting long interdigitating cytoplasmic processes, which frequently contained collections of microtubules and were surrounded by thin, frequently interrupted, basal lamina material. In addition, pinocytotic activity at the cell surfaces and immature junctions joining apposing cellular membranes were identified. No evidence of fibroblastic, fibrohistiocytic, or myofibroblastic differentiation was identified ultrastructurally in the neoplastic cells. Our findings stress the need for ultrastructural examination to adequately classify soft tissue sarcomas. The 2 cases presented illustrate that there are some schwannomas that may be inaccurately classified as malignant fibrous histiocytomas if only light microscopy is used. It seems that only by ultrastructural means is it possible to accurately classify these peculiar neoplasms. PMID- 6293136 TI - Ultrastructure of human tendon sheath and synovium: implications for tumor histogenesis. AB - Normal human tendon sheath and synovium were studied by scanning an transmission electron microscopy. The lining cells of the two tissues appear to be identical ultrastructurally. The most superficial cells (B-cells) possess long cytoplasmic extensions that clothe the membrane surface. Intermingled with deeper B-cells are the so-called A-cells, which have similar cytoplasmic features but lack long processes and instead have many filopodia. the frequent occurrence of intermediate forms indicates that the two cells form part of a morphologic spectrum. Comparison with cells of tumors that have been ascribed to synovium or tendon sheath (synovial sarcoma, epithelioid sarcoma, clear cell sarcoma) do not reveal any close similarities that might support a histogenetic relationship. PMID- 6293138 TI - [Non-visualization of the 12th thoracic vertebra in isotope scintigraphy. Occlusion caused by tumor emboli from anaplastic small cell carcinoma of the lung]. PMID- 6293137 TI - Painful lytic lesion of the left femur in an adult male. PMID- 6293139 TI - [Multifocal glomangioma]. PMID- 6293140 TI - Vasovasostomy in the rat. Improved technique using absorbable microsuture (polyglycolic acid). AB - An improved technique for vasovasostomy in the rat is presented. To eliminate the suture crowds that could cause obstruction of the vasal lumen, one deep raw suture technique had been employed using polyglycolic acid (Dexon) microsuture. A total of 62 vasal anastomoses in 48 male Lewis rats was performed, and follow-up to twelve months found no disruption or constriction at the anastomoses. Despite occasional sperm granuloma formation at the anastomotic sites, all anastomoses demonstrated patency. Dexon appears to be suitable for genitourinary tract surgery, at least, in the rat. PMID- 6293141 TI - Congenital mesoblastic nephroma. AB - For more than a decade congenital mesoblastic nephroma of infancy has been recognized as an entity different from Wilms tumor in its clinical and histopathologic features. However, this tumor is sometimes confused with Wilms tumor and consequently, inadvertent and vigorous therapy compatible with that for Wilms tumor is given. we reviewed the literature and analyzed 90 cases of congenital mesoblastic nephroma in the English and Japanese literature. The importance of early and accurate histopathologic diagnosis of the resected specimen to avoid inappropriate post-operative therapy is emphasized, and the peculiar features of congenital mesoblastic nephroma are characterized. PMID- 6293142 TI - Radioisotope penogram in diagnosis of vasculogenic impotence. AB - A radioisotope technique to estimate penile blood flow is described. The radioisotope penogram is noninvasive and gives a dynamic evaluation of the arterial supply, venous drainage, and blood flow in the corporeal bodies. The penogram is a valuable adjunct in evaluation of patients with vasculogenic impotence. PMID- 6293143 TI - [Anti-relapse therapy of ocular herpes patients with increased sensitivity to herpes simplex virus]. PMID- 6293144 TI - [Detection of T-2 toxin in grains using thin layer chromatography with silica gel (TLC)]. AB - The TLC method of the detection of T-2 toxin in grains is described. A thin layer of silica gel with a sensitivity of 3 mg/kg is used in this method. In comparison with the biological assay for dermal irritability, the TLC method is 30 times less sensitive. However, it has one great advantage over the bioassay: its specificity. It is recommended for practice to use a screening method based on the test for the dermal irritability of rabbits. When the content of toxins is higher, the analytic method can be used for trying to identify the given mycotoxin of the trichothecene group. T-2 toxin was detected in none of the 33 samples of tested grains (14 samples of barley, 7 wheat, 5 rye, 5 oats, 2 samples of maize). PMID- 6293145 TI - Adenovirus in the intestinal epithelium of a foal with prolonged diarrhea. PMID- 6293146 TI - Gastrointestinal neoplasms in nonhuman primates: a review and report of eleven new cases. AB - Reported gastrointestinal neoplasms in nonhuman primates are reviewed, and the clinical and pathologic features of 11 new cases are described. The 11 monkeys had a total of 12 malignant gastrointestinal neoplasms; one had two primary carcinomas, one in the colon and one in the duodenum. Ten of the 12 tumors were adenocarcinomas: two in the duodenum, one in the jejunum, four in the distal ileum or region of the ileocecal valve and three in the large intestine. The remaining two lesions were a histiocytic lymphosarcoma of the stomach and a poorly differentiated sarcoma of the cecum. The 11 animals included nine Macaca mulatta, one Saguinus oedipus oedipus and one Galago crassicaudatus. All were adults and most were aged. There were six females and five males. Clinical signs included progressive weight loss, a palpable abdominal mass and intermittent diarrhea. Grossly, five of the adenocarcinomas were annular, and constricted the intestinal lumen. Microscopically, the carcinomas generally were well differentiated, and two produced mucin in quantities warranting the modifier "mucinous" adenocarcinoma. All tumors were locally invasive and six of nine monkeys with carcinomas had metastases, with the regional lymph nodes the principal site of involvement. PMID- 6293147 TI - Acute gastric dilatation in nonhuman primates: review and case studies. AB - Acute gastric dilatation occurs sporadically in laboratory-housed nonhuman primates. Clinical histories often include chronic drug administration, food restriction, accidental overfeeding, and prior anesthesia. Monkeys may be found dead or may have clinical signs of colic, abdominal distention, and dyspnea. Death in untreated cases is due to impaired venous return and cardiopulmonary failure. Gastric distention with fermented gaseous ingesta and congestion of the abdominal viscera are the predominant lesions. The cause of acute gastric dilatation is unknown, but it probably is multifactorial. Two principal factors seem to be intragastric fermentation associated with Clostridium perfringens, and abnormal gastric function. PMID- 6293148 TI - Adenoviral pancreatitis in rhesus monkeys: current knowledge. AB - Features and pathogenesis of adenoviral pancreatitis in rhesus monkeys were studied with an immunofluorescence staining procedure on tissues from two previously documented cases. Fluorescing adenovirus antigen in epithelial cells of the pancreatic ducts, duodenum, and jejunum suggests that under as yet undefined conditions, a primary adenovirus infection of the gastrointestinal tract ascends to the pancreatic parenchyma via pancreatic ducts. In a retrospective survey, over 3,000 microslides of pancreas taken at necropsy from several species of nonhuman primates (1,002 animals) were studied to determine the incidence of and species susceptibility to adenoviral pancreatitis. Other than the two documented cases from our files, we found comparable lesions in only one rhesus monkey. Adenoviral pancreatitis seems to be a distinct entity in rhesus monkeys, and it should be considered when pancreatitis is found in this species. Our findings also suggest a possible viral cause for some cases of pancreatitis in man. PMID- 6293149 TI - Enteric viruses of nonhuman primates. AB - The phylogenetic relationship of nonhuman primates to man implies that many of these animals could serve as surrogates for studies of diseases of man. Many nonhuman primate species are susceptible not only to viruses of human origin but also to nonhuman primate viruses that are counterparts of viruses of man. All monkeys and great apes do not respond similarly to an antigenic stimulus. Some agents are highly pathogenic for one species and completely innocuous for another. For example, poliovirus causes disease and fatalities in great apes, but picornaviruses given orally cause few lesions in most nonhuman primates. Other enteroviruses (coxsackie-, echoviruses) have caused disease in nonhuman primates. It is difficult to separate viruses into distinct categories according to their anatomic affinities. Many viruses not considered to be enteric may be recovered from the intestinal tract. Adenoviruses, both human and nonhuman strains, which are not considered enteric viruses, nonetheless are recovered frequently from the intestinal tract. Adult animals show little evidence of disease, with the possible exception of diarrhea, after adenovirus infection. Newborns, however, may respond with a fatal pneumoenteritis. Adenovirus may be associated with diseases in organs other than the intestines. The reoviruses, which may be recovered from the intestinal tract, also are generally innocuous. Rotaviruses as pathogens in nonhuman primates are presently under study, and it is suspected that rotaviruses of man may produce experimental disease in nonhuman primates. Production of diabetes by several of the enteric viruses has been suggested but not demonstrated conclusively. PMID- 6293150 TI - Comparative effects of oral administration of trimethoprim/sulphadiazine or oxytetracycline on the faecal flora of horses. AB - A study was carried out on the bacteriological faecal flora of horses before and after oral doses of oxytetracycline or trimethoprim plus sulphadiazine. Administration of oxytetracycline was rapidly followed by large increases in counts of coliforms. Bacteroides and Streptococcus species, the disappearance of Veillonella species, the appearance of Clostridium perfringens type A in large numbers and the accumulation of watery fluid in the rectal contents. These changes were not seen following administration of trimethoprim-sulphadiazine and it was concluded that oral treatment of horses with this combination was unlikely to be accompanied by the hazard of inducing colitis. PMID- 6293151 TI - Picorna-like virus causing hepatitis and pancreatitis in turkeys. PMID- 6293152 TI - Oncoviruses and monoclonal antibody production. PMID- 6293154 TI - Efficacy of some disinfectants against infectious bursal disease virus and avian reovirus. PMID- 6293153 TI - Transmission of feline viral rhinotracheitis. AB - The transmission of feline viral rhinotracheitis (FVR) virus was investigated. Virus could be successfully transmitted between shedding carrier cats and unrelated susceptible kittens, but only if there was intimate contact between them. Studies on the transmission of FVR virus from carrier queens to their kittens showed that although four of 10 queens shed virus in the post partum period, a total of only four kittens from three litters developed a contact infection. All four kittens remained asymptomatic. Two shed for one day only and did not become carriers (as evidenced by corticosteroid treatment) and two shed for 15 days and 25 days and were subsequently shown to have become carriers. None of the remaining kittens tested shed virus. There was no evidence of in utero transmission between FVR-recovered queens and kittens. Passive antibody titres in kittens born to FVR-recovered queens declined to less than 1 in 4 in individual animals by two to 10 weeks of age. Mean titres calculated from a linear regression equation reached less than 1 in 4 and less than 1 in 2 by six and nine weeks of age, respectively. Experiments using a multistage liquid impinger demonstrated that FVR virus was unlikely to be transmitted between cats by aerosol and this was confirmed by the ability of a sentinel cat to withstand virus shedding from carriers over a six month period, although housed in the same air space. PMID- 6293155 TI - Experiments with a homologous, inactivated canine parvovirus vaccine in vaccination programmers for dogs. AB - The significance of canine parvovirus (CPV) infections as a permanent threat susceptible dogs, in particular pups, made the authors develop three liquid homologous inactivated adjuvant CPV vaccines that were compatible with existing canine vaccines and could be incorporated in current vaccination programmes. On vaccine (Kavak Parvo) contained only the CPV component, the second product (Kavak i-LP) also contained two inactivated leptospiral antigens, and the third vaccine (Kavak i-HLP) contained in addition an inactivated canine hepatitis virus. This paper reports on the studies conducted to test the safety and efficacy of the three products. They were used as such and as diluents for freeze dried vaccines containing live attenuated measles, distemper, and hepatitis viruses. The study was performed in a breeding kennel where all dogs were free from CPV antibodies and the nonvaccinated sentinels remained so for the course of the study. All vaccines proved to be safe in dogs of all ages, including pregnant bitches. The efficacy of the CPV component was studied both by monitoring antibody titres for more than a year and by challenge exposure of some dogs to virulent CPV. The results obtained from these studies prove that the CPV component used in the three vaccines can be incorporated as indicated in the recommended canine vaccination programmes. The observations that the inactivated CPV and hepatitis components do induce an active immunity in pups that are still protected by low levels of maternally derived antibodies against these viruses, make those vaccines very suitable in breeding kennels. Additional studies on a comparative basis are being continued in edemically CPV infected breeding kennels to quantify the significance of these observations in these special conditions. PMID- 6293156 TI - Serological examination and egg production of progeny of fowl experimentally infected with egg drop syndrome 1976 virus. AB - Following EDS'76 virus (BC14 virus) infection of breeder chickens by the conjunctival route, vertical transmission occurred in the first week after infection. In the progeny which had been infected with EDS'76 virus by the vertical route, increasing haemagglutination inhibiting (HI) titres to BC14 virus and increasing numbers of birds with HI titres were observed from 3 weeks to 15 weeks of age. Sixty-one per cent of the hens and 77 per cent of the cocks had 2 log HI BC14 virus titres exceeding 4 at an age of 15 weeks. Some birds which han been serologically negative throughout the rearing period, seroconverted between 25 and 28 weeks of age. This phenomenon occurred in hens as well as in cocks. Simulation of stress twice during the laying period by injection of corticosteroid hormone did not increase the number of birds serologically positive to EDS'76 virus. EDS'76 was observed in the group of hens that was vertically infected, since egg production was significantly depressed between 28 and 34 weeks of age. Probably this was mainly the results of a production drop in the hens showing serconversion at 27 or 28 weeks of age. In this group of fowl vertically infected with EDs'76 virus, serologically positive birds appeared to be protected for the greater part to BC14 virus challenge at 50 weeks of age, while negative birds seemed to be fully susceptible. Chicks hatched from eggs collected in the third and fourth week after infection of the dams had maternal antibodies. Fertility and hatchability of apparently normally shelled eggs seemed not to be affected after BC14 virus infection of the dams. Intensive contact with contaminated faeces is probably an indispensable condition for lateral transmission of the virus. PMID- 6293157 TI - Serum antibody responses of neonatal and young adult pigs to transmissible gastroenteritis coronavirus. AB - Serum titers of virus-neutralizing (VN) antibody were 10 to 16 times higher in neonatal pigs than in young adult pigs, after single oral doses of virulent transmissible gastroenteritis virus (TGEV). To determine the reason for this higher response, sera from neonatal and young adult pigs, 18 to 21 days after exposure to TGEV, were collected and assayed for VN antibody by plaque reduction. In addition, sera of VN-positive and VN-negative neonatal pigs were analyzed for immunoglobulin classes by radial immunodiffusion technique. The competence of neonatal pigs to produce VN antibody with increased IgG levels was demonstrated. The higher antibody response seen in neonatal pigs, when compared to sera of young adult pigs, may be attributed to the increased replication of TGEV in the intestinal tracts of neonatal pigs or to the lack of other immunogens that may interfere or compete with the production of specific antibody. PMID- 6293158 TI - Immunohistochemical study of the immunoglobulin classes of the plasma cells in papillary syringadenoma. PMID- 6293159 TI - Oestrogen receptors in human breast cancer. Problems of correlation with histopathological features. AB - Histopathological factors which might explain inconsistency in published data attempting to correlate oestrogen receptor content (ER) and pathological features in primary breast tumours have been investigated in 194 cases. It was found, that unequal assessment of tumour type and of histological grading between observers is one important factor. In terms of grading, however, heterogeneity of growth pattern within the same tumour seems to be of greater significance. No significant correlation was found between histological type of tumour and ER content. However, a trend towards a correlation between the extent of tubule formation (as an indication of differentiation) and ER content was observed. PMID- 6293161 TI - Ultrastructure of a virilizing ovarian Leydig-cell-tumor. Hilar cell tumor. AB - A case of virilizing ovarian hilus cell tumor (Leydig-cell tumor) in a 37 year old female was studied by light and electron microscopy. The ultrastructural features of this rare and almost always benign tumor are compared with those reported in the literature and with findings in normal and neoplastic interstitial cells of the testis. Tubulovesicular hyperplasia and formation of whorl structures of the endoplasmatic reticulum together with the presence of exocytosis vesicles on the cell surface may be the morphological manifestation of endocrine activity of the tumor. The identity of ultrastructural and optical diffraction characteristics of the crystal inclusions in both cells (hilar and testicular interstitial) favours the assumption of an homology of both cells and their neoplasms. PMID- 6293160 TI - Epithelioid sarcoma. Enzyme histochemical and ultrastructural study. AB - A case of epithelioid sarcoma was studied by electron microscopy and by light and electron microscopic enzyme histochemistry comparing with several control soft tissues. In addition to previously reported ultrastructural features, such as abundant 10 nm cytoplasmic filaments, desmosome-like cell junctions and small cystic spaces surrounded by filopodia or microvilli of the tumor cells, we encountered 10 nm cytoplasmic filaments showing electron dense condensation with a concentrically oriented or whorled pattern and a finger-print-like arrangement and 5'-nucleotidase activity of tumor cell membrane. Among the control soft tissues, 5'-nucleotidase activity was found only in synovial and endothelial cells. Both tumor and synovial cells showed no activity of adenosine triphosphatase, while marked activity of the enzyme was found in endothelial cells. These results support the concept that epithelioid sarcoma is derived from mesenchymal cells undergoing differentiation toward synovial cells during neoplastic transformation. PMID- 6293162 TI - Synovial sarcoma arising in an anatomical bursa. AB - In previous studies, the origin of synovial sarcoma directly from synovium has not been satisfactorily established. This case report describes the light and electron microscopic features of a biphasic synovial sarcoma occurring within the popliteal fossa. At surgery, a cystic mass was identified in relationship to the semitendinosus tendon at the anatomical site of the semitendinosus bursa. The tumour originated from the inner surface of the bursa as multiple papillary projections with no evidence of extension beyond the capsule of the bursa. Portions of the synovial surface were hyperplastic but otherwise normal. The findings indicate that biphasic synovial sarcoma can arise directly from synovium and support the hypothesis of a mesenchymal histogenesis for this tumour. PMID- 6293163 TI - The effect of influenza vaccination on the circulation of viruses in the respiratory tract of apparently healthy schoolchildren aged 7 to 14 years. AB - Virus isolation attempts were made with 500 nasopharyngeal secretion samples from apparently healthy schoolchildren aged 7-14 years, vaccinated by nasal route with an inactivated influenza vaccine and with 500 samples from unvaccinated schoolchildren of the same age. The number of virus isolates obtained was much smaller in the former than in the latter group (13 as against 50 virus isolates). The role of influenza vaccination in the protection of schoolchildren against respiratory tract infections is discussed. PMID- 6293164 TI - Virus--tumor host cell relationships. In vivo cocultivation of adenovirus type 5 and of SV40 in mouse Ehrlich ascites carcinoma. AB - Mouse Ehrlich ascites carcinoma (EAC) cells proved to be a semipermissive substrate for in vivo cultivation of adenovirus 5 (Ad5) and SV40. The multiplication of SV40 in EAC cells was facilitated by the coinfection with Ad5. As demonstrated by ID and EID reactions, the virus progens isolated at the first passage after the mixed infection of EAC cells with Ad5 and SV40 possess an antigenic mosaic with fractions characteristic of the parental viruses and of the cell substrate in which they had cultivated in vitro and in vivo. The progens gave positive seroneutralization and complement fixation reactions only with antiserum to SV40. PMID- 6293165 TI - Experimental infection of soybean and transmission of Sendai virus through the seeds. AB - Sendai virus can multiply in the soybean and it can be serially transmitted over at least three passages. The presence of the virus was made evident in leaf homogenates from infected plants after several blind passages in egg chorioallantoic membrane fragments. The virus can be transmitted through the seeds of experimentally infected soybean plants and reisolated from leaves and roots. PMID- 6293166 TI - Presence of antibodies to human cytomegalovirus in patients with different forms of cancer and in other categories of subjects. AB - Complement fixing (CF) antibodies to the AD--129 strain of human cytomegalovirus (HCMV) were detected in patients with different forms of cancer, as well as in blood donors, in a proportion of 16.6% and 73.2%, respectively. The prevalence of CF antibodies to HCMV, strain AT--129, in the population of Romania is similar to that reported in other countries. PMID- 6293167 TI - Herpes virus infection and uterine cervix carcinoma. General considerations, personal experimental data and suggestions for a specific antiherpes adjuvant therapy. AB - The authors review some data of the literature and results of personal investigations that point out a relationship between human herpes simplex virus infection and the etiopathogeny of uterine cervix carcinoma. On the ground of these data the authors suggest a "specific antiherpes" adjuvant therapy in this form of human cancer. PMID- 6293168 TI - Prevalence of hemagglutination-inhibiting antibodies to BK virus in patients with endemic Balkan nephropathy. PMID- 6293169 TI - Phosphonoacetic acid-induced morphological changes of herpesvirus growth in vitro. AB - Thin-section electron microscopy of Herpesvirus hominis type 1 (HV-1)-infected HEp-2 cells exposed to 100 micrograms/ml phosphonoacetic acid (PAA) revealed not only a spatial restriction of virus multiplication to a few spheroid bodies in the nucleus, but also a delay in viral development, resulting in the formation of unusual HV-1 particle rosettes around the spheroid bodies. HV-1-infected cells were lacking, however, the small tubular structures known to accumulate at increased rates in the presence of PAA, in HV-2-infected cells. PMID- 6293170 TI - Electron optic aspects of a murine cytomegalovirus strain grown in R1 mouse kidney cell cultures. AB - The electron microscopic study of a mouse kidney cell line (R1) infected with a murine cytomegalovirus strain isolated from a micromammal captured in the area of endemic Balkan nephropathy demonstrated the presence of multiple types of virus particles, in different phases of replication. Nucleocapsids with a partially or almost totally empty centre were predominant. The small number of particles with an electron-dense core and double envelope points to a low yield of mature infectant particles. PMID- 6293171 TI - Preliminary data on the survival length of enteroviruses in nonalcoholic beverages. Inactivating action of some medicinal herb infusions. AB - Vaccinal poliovirus type 1, Coxsackie B3 virus and echovirus type 1 survived in mineral water, camomile infusion and coffee for over 20 days. In contrast, Chinese tea, lime and mint infusions had a possibly transient inactivating effect on the same enterovirus strains. There was a direct relationship between the concentration of the infusions and their inactivating action. The vaccinal poliovirus was more sensitive than Coxsackie and echoviruses to the inactivating action of the infusions. PMID- 6293172 TI - Common model of normal cell differentiation and oncogenesis based on the mutation -recombination effects of transposable genetic elements and of provirus states. AB - Cell differentiation would result from a sequence of mutation--recombination events that occur in quantal cell cycles, initiating cell lineages. The cytogenomic proviruses--transposable elements ensemble would represent the mutation--recombination equipment of differentiation. Carcinogenesis, as a consequence of multicausally induced mutation--recombination events, would likewise occur in a particular quantal cell cycle resulting in the malignant cell lineage. Superposing the environmental causes of malignancy over those of genetic diseases, the model suggests common prevention methods. PMID- 6293173 TI - Frequent intragenic transcription termination within the N gene of vesicular stomatitis virus. PMID- 6293174 TI - Partial characterization of temperature-sensitive mutants of pseudorabies virus. PMID- 6293175 TI - Comparison of the physical and genetic maps of pseudorabies virus shows that the genetic map is circular. PMID- 6293176 TI - Identification of HTLV p19 specific natural human antibodies by competition with monoclonal antibody. PMID- 6293177 TI - Complementation of a tsa mutant and replication of a recombinant DNA carrying the viral ori region in mouse cells transformed by polyoma virus. PMID- 6293178 TI - Cloned endogenous mouse mammary tumor virus DNA is biologically active in transfected mouse cells and its expression is stimulated by glucocorticoid hormones. PMID- 6293179 TI - Nucleotide sequences of herpes simplex virus type 1 (HSV-1) affecting virus entry, cell fusion, and production of glycoprotein gb (VP7). PMID- 6293180 TI - Transformation of lymphocytes by Epstein-Barr virus requires only one-fourth of the viral genome. PMID- 6293181 TI - Genomic and receptor attachment differences between mengovirus and encephalomyocarditis virus. PMID- 6293182 TI - Induction of mutations within the host cell genome by partially inactivated herpes simplex virus type 1. PMID- 6293183 TI - Isolation of poliovirus variants resistant to and dependent on arildone. PMID- 6293184 TI - Avian endogenous retroviral envelope glycoprotein is assembled in two structural complexes of gp85 and gp37 subunits. PMID- 6293185 TI - Production and characterization of a monoclonal antibody to the N protein of vesicular stomatitis virus (Indiana serotype). PMID- 6293186 TI - Proteus mirabilis phage 5006M: restriction maps of genome in relation to headful packaging. PMID- 6293187 TI - Activation of cellular stress protein genes by herpes simplex virus temperature sensitive mutants which overproduce immediate early polypeptides. PMID- 6293188 TI - A study of the antiviral mechanism of action of 2-deoxy-D-glucose: normally glycosylated proteins are not strictly required for herpes simplex virus attachment but increase viral penetration and infectivity. PMID- 6293189 TI - Restriction analysis and molecular cloning of endogenous murine leukemia virus specific DNA sequences of the mouse genome. PMID- 6293190 TI - Ner, a cro-like function of bacteriophage Mu. PMID- 6293191 TI - Characterization of papillomaviruses isolated from cutaneous fibromas of white tailed deer and mule deer. PMID- 6293192 TI - A fraction of Epstein--Barr virus virion DNA is methylated in and around the EcoRI-J fragment. PMID- 6293193 TI - Intracellular cleavage of an SSV coded gag-related protein. PMID- 6293194 TI - Translation of capped and uncapped VSV mRNAs in the presence of initiation factors from poliovirus-infected cells. PMID- 6293195 TI - SV40 early mutants that are defective for viral DNA synthesis but competent for transformation of cultured rat and simian cells. PMID- 6293196 TI - Leukotrienes: a novel group of biologically active compounds. PMID- 6293197 TI - [Angioimmunoblastic lymphadenopathy with paraproteinemia and the presence of intracytoplasmic crystalloid inclusions with transformation into malignant lymphoma]. PMID- 6293198 TI - [Aspects of military medical expertise in diseases of the lumbosacral portion of the peripheral nervous system]. PMID- 6293199 TI - Red blood cell cold agglutinins and B lymphocyte cytotoxins in breast cancer sera. AB - Cytotoxic activity towards B-lymphocytes in sera from individuals with breast carcinoma was related to red blood cell cold agglutinin levels and IgM values. The control group consisted of 32 females with a similar age distribution. Significant increases in red blood cell cold agglutinin levels were found in the group of cancer patients and the group with the histological category of mucin producing carcinoma with lymphocyte cytotoxins (p less than 0.025, respectively), in comparison to the control group and cancer patients without lymphocyte cytotoxins. Serum IgM concentrations were significantly increased in the group of cancer patients (p less than 0.025) and the group with mucin-producing carcinoma (p less than 0.01) with lymphocyte cytotoxins, in comparison to cancer patients and controls without lymphocyte cytotoxins. The lymphocyte cytotoxins were completely or partially adsorbed by red blood cells in 58% of the positive sera. These results suggest that red blood cell cold agglutinins constitute a portion of cytotoxic anti-B lymphocyte antibodies. At present it is unknown whether the lymphocytotoxic antibodies in breast cancer patients are of importance in immunologic host defenses. PMID- 6293200 TI - [Interrelation between the age of the patient, the function of his regulatory systems and myocardial infarct]. PMID- 6293201 TI - [Treatment of peripheral nervous system and locomotor apparatus diseases at Gornaia Tisza sanatorium]. PMID- 6293202 TI - [Indirect hemagglutination reaction as a rapid method of influenza serodiagnosis in natural biocenoses]. PMID- 6293203 TI - [Transformation and malignant degeneration of newborn hamster cells inoculated with herpes simplex virus type 2]. PMID- 6293205 TI - [Serum haptoglobin and 5'-nucleotidase levels in patients with acute viral hepatitis with low and high bilirubin levels]. PMID- 6293206 TI - [Differential diagnosis of "P-pulmonale" by the beta-adrenergic blockade test]. PMID- 6293207 TI - [Current theories on peripheral diabetic neuropathy]. PMID- 6293204 TI - Mast cells in allergic diseases and mastocytosis. AB - Mast cells with their stores of vasoactive and chemotactic mediators are central to the pathogenesis of allergic diseases. The cross-linking of receptorbound IgE molecules on the surface of mast cells initiates a complex chain of events, including calcium ion influx, phospholipid methylation and turnover and cyclic nucleotide metabolism, ultimately resulting in the release of mediators of immediate hypersensitivity. These mast cell mediators are important in smooth muscle reactivity, in the recruitment of eosinophilic and neutrophilic leukocytes and in the generation of secondary chemical mediators. Histologic evidence of mast cell degranulation, biochemical evidence of mast cell mediators in blood and tissues and clinical evidence of signs and symptoms reproducible by these mediators have strongly supported the crucial role of mast cells in asthma, urticaria, anaphylaxis, rhinitis and mastocytosis. Because of their unique location at host environment interfaces, mast cells may both participate in allergic diseases and promote homeostasis. PMID- 6293208 TI - [Virus demonstration in liver cell carcinoma]. PMID- 6293209 TI - Metabolism and subsequent covalent binding of benzo[a]pyrene to macromolecules in gonads and liver of ripe english sole (Parophrys vetulus). AB - 1. Ripe English sole (Parophrys vetulus) force-fed [3H]benzo[a]pyrene, contained 1% of the dose in liver, 0.2% in ovary and 0.1% in testis, after 24 h. No significant change occurred in levels of radioactivity from 24 to 168 h. 2. Gonads and blood contained substantially larger proportions of unchanged benzo[a]pyrene (15-37% of tissue radioactivity) and organic solvent-soluble metabolites (6-35%) than did liver and bile. 3. T.l.c. revealed the presence of phenols, quinones, 7,8-dihydro-7, 8-dihydroxy- and 9,10-dihydro-9,10-dihydroxy benzo[a]pyrene in liver and gonads. 4. A small proportion (less than 10%) of the radioactivity in liver and gonads was present as glucuronides and sulphates; bile contained a higher proportion (ca. 20%) of total radioactivity as glucuronides and sulphates. 5. Benzo[a]pyrene intermediates were covalently bound to liver proteins and DNA, and to a lesser extent to gonadal proteins (male and female fish) and gonadal DNA (confirmed for testis only). PMID- 6293210 TI - Species differences in the effects of 1,1-di-(4-chlorophenyl)-2-chloroethylene (DDMU) and 1,1-di-(4-chlorophenyl)-2,2-dichloroethylene (DDE) on glutathione levels in isolated hepatocytes from Japanese quail and rat. AB - 1. Hepatocytes have been isolated from adult Japanese quail in high yields (26-40 X 10(6) cells/g) with 95% viability. Variation in the collagenase and hyaluronidase levels and incubation time used in the isolation procedure affected the yield and viability of the hepatocytes. Quail hepatocytes were more stable in a standard nutrient medium than those of the rat. 2 1,1-Di-(4-chlorophenyl)-2 chloroethylene (DDMU), a metabolite of DDT, depleted quail hepatic GSH levels both in vitro and in vivo, but had no effect on rat hepatocytes. DDMU (0.1 mM) depleted GSH levels in the quail to the same extent as diethyl maleate (a known GSH-depleting agent) (0.04 mM) but the latter acted more rapidly. 3. Pretreatment of quail in vivo with DDMU or phenobarbitone, known inducers of the hepatic mixed function oxidases, resulted in faster depletion of GSH when the hepatocytes were incubated subsequently with DDMU in vitro. 4. In contrast, 1,1-di-(4 chlorophenyl)-2,2-dichloroethylene (DDE), another metabolite of DDT, did not deplete GSH levels in the quail but did cause some reduction in the rat. Phenobarbitone pretreatment had no effect on GSH depletion by DDE in vitro in quail hepatocytes but enhanced GSH depletion in rat hepatocytes. PMID- 6293212 TI - Decrease in essential fatty acid content of edible fats during the frying process. AB - Degradation of sunflower oil, rapeseed oil, and lard during the frying operation was investigated by studying the loss of essential fatty acids compared to the accumulation of decomposition products. Linoleic and/or linolenic acid concentration was measured by GLC, and for detecting decomposition products determination of polymer content by GPC was chosen. Twelve laboratory experiments with different heating or frying conditions were run aimed at modelling practice and studying the effects of certain factors. The results indicated that loss of essential fatty acids being a parallel process to the accumulation of breakdown products is suitable both for detecting the decrease in nutritive value and for quality assessment of used frying fats. It was also found that the rate of deterioration is considerably affected by the nature of fat and the frying parameters. Study of the relationship between polymer content and the decrease in essential fatty acid concentration (using data from 110 samples) showed that high, linear correlation depending on the nature of fat can be found. Correlation coefficients and equations of regression lines were calculated. PMID- 6293211 TI - Recent advances in the pathogenesis and nutritional treatment of chronic uremia. PMID- 6293213 TI - [Leser-Trelat syndrome, erythema anulare centrifugum Darier, breast cancer and basaliomas in a patient]. AB - A 46 year old woman had suffered from Erythema anulare centrifugum Darier (EAC) since 20 years. During a rush of the erythema numerous seborrhoic keratoses appeared (Leser-Trelat sign/LTS). Clinical examination revealed a ductular carcinoma of the breast. During the last two years, both types of lesions increased although the tumor had been treated. In addition, the patient developed several basal cell epitheliomas at forehead and chest. PMID- 6293214 TI - Host-parasite interface of the fluke Collyriclum faba (Bremser in Schmalz, 1831) as revealed by light and electron microscopy. AB - Light and electron microscopy of Collyriclum faba reveals that the dorsal tegument is highly convoluted, containing regular groups of between one and five spines. The thinner ventral covering has a less spiny surface with shallow infoldings. The lining of the capsule is characterized by numerous layers of collagen reflecting a strong cellular response by the host. PMID- 6293215 TI - Crystalline inclusions in the subtegumental cells of the adult lung fluke, Paragonimus westermani. PMID- 6293216 TI - [Superoxide dismutase activity and the membrane formation of superoxide radicals in tumorous and normal tissues]. PMID- 6293218 TI - Excretion of nitrate and nitrite by the pig as influenced by dietary fibre levels and microbial activity in the digestive tract. PMID- 6293217 TI - [Regulation of the phosphodiesterase activity of cyclic nucleotides (new endogenous regulators)]. PMID- 6293219 TI - [Multiple unilateral tumors of the salivary parotid gland]. PMID- 6293220 TI - [Results of pneumonectomy for bronchial carcinoma]. AB - 615 patients suffering from lung cancer underwent pneumonectomy from 1966 to 1975 (standardized operation: 379; radical pneumonectomy: 236). Evidently the rate of postoperative insufficiency of the bronchial stump and pulmonary embolism was on the increase in cases of radical surgery. The postoperative mortality rate (including 30th postoperative day) was double as high in the radical group as in standardized operations. The 5 years survival rate following radical surgery came up to 13.1% (standard group 27%). This justifies radical pneumonectomy because there is no alternative left for the patients afflicted. PMID- 6293222 TI - [Changes in chrysotile in vivo]. AB - In an animal experimentation rats received a single intratracheal injection of 2 mg chrysotile per animal. Quantitative energy dispersive x-ray microanalysis (EDXA) and selected area electron diffraction (SAED) patterns of the chrysotile fibres, obtained from lung tissue, were performed with the aid of a transmission electronmicroscope (TEM). During a period of four months the chemical and physical instability of asbestos fibres were investigated. The results of microanalysis were compared with standard chrysotile A, UICC as a reference material. The magnesium-leakage of chrysotile fibres and in the diameter range below 0.1 micrometer was found to be time-depending and increased during the period of the experiment. Opposite to the contents of Ca, the Fe-concentration was not significantly higher. After two months already, a greater number of depleted chrysotile fibres could be analysed. If leakage of magnesium exceeded 80 percentage, the typical single crystal electron diffraction patterns of the chrysotile could not be obtained furthermore. EDXA and SAED are inevitable techniques of identify fibres, which been altered during the period of deposition in animal lungs. There was no evidence to suggest the formation of asbestos bodies during the period of experimentation. PMID- 6293221 TI - [Scanning electron microscopical investigations on the respiratory epithelium of the Syrian golde4n hamster. V. Dose-related in vitro effects of benzo(a)pyrene]. AB - The biological effects of in vitro exposure of fetal hamster tracheal explants to three dose levels of benzo(a)pyrene (BaP:25, 50, 100 microgram/ml medium) in 0.5% dimethylsulphoxide (DMSO) were studied by light and scanning electron microscopy. A dose-related difference was seen in a quality of the observed pathological alterations in the specimens examined after 28 days of culture. Whilst in the explants treated with 50 or 100 microgram/ml BaP the respiratory epithelium revealed necrotic areas and extended metaplastic alterations, explants exposed to 25 microgram/ml BaP showed normal differentiation in some areas of the respiratory epithelium. In explants treated with the highest dose (100 microgram/ml BaP), hyperplastic foci were seen in the epithelium and tended to proliferate towards the tracheal lumen. These observations show similarity with early neoplastic alterations. PMID- 6293223 TI - [Poxvirus infection in a domestic cat]. PMID- 6293224 TI - [Toxigenicity conversion by pertussis phages in Bordetella parapertussis]. AB - For the first time toxigenicity conversion in B. parapertussis induced by B. pertussis phages was discovered. The clones of B. parapertussis recipient strain No. 17903 used in this study were subjected to lysogenization with 4 B. pertussis phages; as a result, 95% of these clones became immune to the repeated phage infection, developed spontaneous phage production and showed toxic properties (lethal toxicity due to the action of thermolabile and thermostable toxins) characteristic of the donor strains from which B. pertussis phages had been obtained. Differences in the degree of toxicity shown by the converted strains were determined by means of the spleen index. The convertants thus obtained did not possess protective potency. PMID- 6293225 TI - [Quantitative evaluation of the content of antibodies to hepatitis of A virus in serial immunoglobulin preparations]. AB - A total of 156 samples of several lots of immune serum globulin (ISG) were tested for antibodies to hepatitis A virus (anti-HAV). In this study the blocking variant of the solid-phase enzymoimmunological method was used. The samples under test were the production of 12 manufacturing enterprises in the USSR and I enterprise in Czechoslovakia. All the samples under test were found to contain anti-HAV within the limits from 2500-5000 to 20000-40000. The samples produced by different manufacturing enterprises showing no essential differences in the content of anti-HAV. Most preparations (93.6%) contained antibodies at levels of 5000-20000. The ISG proposed as the international standard preparation had a titer of 8000-10000. No differences in the content of anti-HAV were detected in comparing ISG whose shelf life expired 4 years before the test with ISG whose shelf life was to expire 3 years later. The results obtained in this study are discussed in connection with the immunoprophylaxis of viral hepatitis A. PMID- 6293227 TI - Basic function in the nervous system - a unified theory. PMID- 6293226 TI - [Modulating functions of yeast RNA]. PMID- 6293228 TI - [Perinatal development of glucose-6-phosphatase and fructose-1,6-diphosphatase activities in pig liver]. AB - The activity of glucose-6-phosphatase (G6Pase) and fructose-1,6-bisphosphatase (FDPase) was determined in the homogenate of the liver of 69 pig fetuses during the last third of gestation (80th to 114th day), 47 piglets from birth to 4 weeks old (suckling period) and to slaughter pigs. G6Pase is evident in fetal liver at an early date and raises steadily during gestation. In newborn piglets, the enzyme activity increases rapidly during the first hours of life and remains at this high level during the first week of life. Afterwards the enzyme activity returns to birth level, which exists also in pigs at slaughtering. The activity of FDPase is constant during the fetal period. After birth enzyme activity rises at a lower rate than the G6Pase during the first week of life. This level remains constant during the suckling period and increases thereafter until the time of slaughtering of pigs. The role of hormones in the perinatal development of these enzymes is described. Probably, thyroxine causes the prenatal increase of the activity of both the enzymes. The rapid postnatal rise of G6Pase activity may be induced by the high level of hydrocortisone at parturition, and furthermore, glucagon may have a permissive effect. PMID- 6293229 TI - The application of a sepharose bead immunofluorescence assay and a solid-phase radioimmunoassay to the bovine leukemia virus system. AB - Several fluorescence assays with bovine leukemia virus (BLV) conjugated to activated Sepharose 4B were used for the detection of BLV and anti-BLV antibodies. These tests were compared with a solid-phase radioimmunoassay and found to be in the same sensitivity range. Sepharose bead immunofluorescence assay and solid-phase radioimmunoassay can be applied to the diagnosis of BLV infection in cattle. PMID- 6293230 TI - Sputum cytology for the diagnosis of carcinoma of the lung. AB - Of 400 consecutive patients with histologically proven carcinoma of the lung, one to six sputum samples (mean, 2.8) were examined cytologically; 60% also had histologic examination of paraffin-embedded material. The overall sensitivity of sputum cytology was 0.58. The sensitivity increased from 0.37 to 0.57 when three samples rather than one were examined; it increased by only another 0.01 when four to six samples were studied. The examination of paraffin-embedded material yielded another 0.075 increase in sensitivity. A multiparametric study, including diameter, cavitation, site and histologic type of the pulmonary tumors, showed that sputum cytology was particularly significant for neoplasms of the left upper lobe and that the sensitivity related to the histologic type was not independent of the site, diameter and cavitation. The overall cytologic typing accuracy was 0.77, with a range from 0.20 to 0.96. The majority of the diagnoses at variance with histology and the unclassified malignant epithelial tumor cells were found to be associated predominantly with carcinoma of the large-cell type and with poorly differentiated adenocarcinomas. PMID- 6293231 TI - Cytologic diagnosis of pulmonary carcinoma on bronchoscopic brushing material. AB - The material obtained by brushing procedures under visual control from 370 consecutive patients with histologically confirmed primary carcinoma of the lung who underwent bronchoscopy was examined cytologically. The sputum of 276 of these patients was also investigated. The overall sensitivity of the bronchoscopically obtained cytology was 0.67; however, the values were 0.78 and 0.28, respectively, for the cases with and without visible lesions of the bronchi. Sputum examination enhanced the sensitivity to 0.79 overall and to 0.84 and 0.61, respectively, for the two groups, with a greater benefit for the group with negative bronchoscopy. The overall cytologic typing accuracy for 252 cases was 0.66, with a range of 0.19 to 0.86, depending on the histologic type. Typing failures were essentially related to adenocarcinomas and large-cell carcinomas. PMID- 6293232 TI - A characterisation of cyclic AMP release from monolayer cultures of normal human thyroid cells. AB - The thyrotrophin (TSH)-dependent and time-related release of cyclic AMP has been characterised in primary monolayer cultures of normal human thyroid cells. Accumulation of cyclic AMP within the incubation medium was detectable within 1 h of exposure of cultures to 5 mU TSH/ml, and increased throughout of subsequent 15 h incubation period, final levels attained being consistently in excess of the corresponding intracellular cyclic AMP levels. Accumulation of cyclic AMP in the incubation medium was dependent on TSH dose, for both short (1 h) and prolonged (16 h) incubations. Moreover, after incubation for 16 h, cyclic AMP levels in the incubation medium were significantly (P less than 0.01) in excess of intracellular levels for each dose of TSH tested above 0.2 mU/ml. In the absence of TSH, accumulation of cyclic AMP in the incubation medium remained low, after both 1 h and 16 h incubation periods. A consideration of these observations suggests that a bioassay based upon the cyclic AMP content of incubation medium samples should provide a more precise detection system for thyroid stimulators than those measuring the intracellular cyclic AMP response, and this has been demonstrated for a cell preparation in which the intracellular response to TSH was minimal. PMID- 6293233 TI - Somatostatin-28 like immunoreactivity in normal and tumour tissue from duodenum and pancreas. AB - Radioimmunoassay using labelled somatostatin-14 revealed that components of somatostatin-28 antisera cross-reactive to somatostatin-14 were removed by absorption of somatostatin-28 antisera with sepharose 4B-somatostatin-14. Indirect immunofluorescence techniques using specific antisera against somatostatin-28 were carried out in normal pancreas, duodenum, a somatostatinoma in the duodenum, and pancreatic tumour cells containing somatostatin-14 positive cells, in order to establish if somatostatin-28 is present in normal and pathological tissues. Somatostatin-28 like immunoreactivity was present in pancreatic islets cells and in the epithelial cells of the duodenum as well as in the duodenal somatostatinoma and in pancreatic tumour cells. Furthermore, cells reacting with specific antisera against somatostatin-28 were identical to those with somatostatin-14 antisera in normal and pathological tissues. The findings suggested that somatostatin-28 like immunoreactivity may be constantly present in the tissues where somatostatin like immunoreactivity was detected using somatostatin-14 antisera. However, further studies were necessary to clarify whether somatostatin-28 and somatostatin-14 were independently present in these tissues, in other words, whether somatostatin-14 may be produced from somatostatin-28 or not, since somatostatin-14 antisera had cross-reactivities to somatostatin-28. PMID- 6293234 TI - [Bone marrow culture, cytochemistry and electron microscopy in agar in patients with preleukemic syndrome and aplastic anemia]. AB - Thirty-seven patients with chronic cytopenia were studied using a CFU-c assay in agar. On the basis of the growth pattern three types of preleukaemic syndrome (PL) and two types of aplastic anaemia were distinguished. Further evaluation of the bone marrow dysfunction was attempted with a combined application of cytochemistry and electron microscopy for the morphologic study of cells proliferating in vitro. Well-defined maturation defects in the growing cells from the bone marrow of patients with PL were demonstrated with cytochemical stainings performed in agar. These results were supported by electron microscopic findings of Auer-body-like inclusions in "statu nascendi" in the vacuoles of preleukaemic cells. On the basis of our results a high risk group of PL for development of overt leukaemia and a group of patients with a grave prognosis in aplastic anaemia were distinguished. The data obtained are relevant for the clinical diagnosis and prognosis of patients with cytopenias. PMID- 6293236 TI - Ketamine and sympathetic mechanisms in cardiac and smooth muscle. AB - The effects of ketamine on the response of rat isolated cardiac and smooth muscle to electrical stimulation of intramural sympathetic nerves and to exogenous norepinephrine are reported. Low concentrations of ketamine (1 x 10(-5) to 5 x 10(-5) mol l-1) significantly potentiated, whilst high concentrations (2 x 10(-4) and 4 x 10(-4) mol l-1) significantly depressed the positive inotropic response of left atria to sympathetic nerve stimulation. Ketamine 5 x 10(-5) mol l-1 caused a leftward shift of the concentration-response curves of left atria to norepinephrine (threefold increase in potency) and isoproterenol (twofold increase in potency). Spontaneously beating right atria were slowed in a concentration-dependent manner by ketamine (5 x 10(-5) to 4 x 10(-4) mol l-1). Ketamine 1 x 10(-4) mol l-1 potentiated the positive chronotropic response of right atria to low frequency (2 and 5 Hz) sympathetic nerve stimulation, whilst ketamine 4 x 10(-4) mol l-1 depressed the response to all stimulation frequencies. Ketamine 3 x 10(-5) mol l-1 significantly potentiated the amplitude of the response of both hepatic portal vein and vas deferens to field stimulation of sympathetic nerves, and to exogenous norepinephrine. In the presence of reuptake blockade of norepinephrine by pancuronium 1 x 10(-4) mol l-1, ketamine caused no further potentiation of the response of the vas to nerve stimulation. PMID- 6293235 TI - A review of succinylcholine-induced apnea. PMID- 6293237 TI - [Personal experience with the surgery of leprosy in Yaounde' (Cameroon)]. PMID- 6293238 TI - [15 years of surgery of leprotic nerves. Results of 644 interventions as a function of advanced forms]. PMID- 6293239 TI - Cytomegalovirus antibodies in epileptics receiving diphenylhydantoin. AB - Cytomegalovirus (CMV) antibodies were determined by indirect haemagglutination in 53 epileptics receiving long-term diphenylhydantoin (DPH) therapy and in 53 matched controls. Absorption of serum IgG, IgA and IgM performed in 12 sera showed that the antibodies were of the IgG class. A decreased incidence of high CMV antibody titres (greater than or equal to 320) was found in epileptics (11.3%) compared with controls (34.0%) (0.02 greater than P greater than 0.01), whereas antibody titres greater than or equal to 40 were found in 37.7% of epileptics and in 45.3% of controls (n.s.). The CMV antibody incidence and titre range were similar in patients with symptomatic (37.9%) and idiopathic (37.5%) epilepsy, suggesting that the DPH treatment was responsible for the decreased antibody occurrence in patients. No correlation between CMV antibody titres and the serum immunoglobulin levels or the DPH concentration or clearance could be established in the epileptics. PMID- 6293240 TI - The course of alcoholic-nutritional peripheral neuropathy. AB - 63 patients with alcoholic-nutritional peripheral neuropathy were given neurologic, electrophysiologic and nutritional examinations. 24 of these patients were reexamined later in the course of their disease, after from 2 to 72 months (mean 33). Alcoholic-nutritional neuropathy appeared and worsened after bouts of heavy alcohol intake and malnutrition. Initially it was sensory and symmetric in character, with prominent involvement of the posterior tibial nerves. With repeated attacks it became more proximal, more motor, and associated with more severe slowing of nerve conduction velocity. 11 of the patients were able to stop drinking alcohol. Initial subjective improvement was seen within the first week or two, but substantial improvement was not seen for 5 to 6 months. Most leg motor nerve velocity improved at a mean rate of increase of 0.12 M/sec per abstinent month. Large motor units and slowed nerve conduction persisted in "cured" patients. The largest motor units detected in the legs grew, despite alcohol intake. PMID- 6293241 TI - Surgical treatment of strabismus. A retrospective investigation of results of surgical treatment of horizontal strabismus. PMID- 6293243 TI - Transactions of the XXIst Congress of the Scandinavian Oto-Laryngological Society. Turku, August 16--19, 1981. PMID- 6293242 TI - Virus particles in the cochlear spiral ganglion of guinea pigs. AB - All examined spiral ganglions of several guinea pig populations from different breeds showed intracytoplasmic viruses in some granular spiral ganglion cells. According to their localization and morphology we classify these viruses with the oncorna group. This is not in agreement with the classification of other authors. Apparently there is a world-wide latent viral infection in guinea pigs. Beside the virus particles we found an accumulation of lysosomes which indicate a local increased lysosomal activity of the infected ganglion cells. Further influences on the infected cells can neither be demonstrated nor denied. PMID- 6293244 TI - Inflammatory fibrous histiocytoma of the urinary bladder: clinicopathological report of a case. PMID- 6293245 TI - Fatty acid analysis for differentiation or Bordetella and Brucella species. AB - The fatty acid composition of Bordetella pertussis (13 strains), B. parapertussis (3 strains), B. bronchiseptica (6 strains), Brucella abortus (6 strains), B. melitensis (4 strains) and B. suis (5 strains) was determined. Both genera contained straight-chain saturated and mono-unsaturated acids as well as cyclopropane substituted isomers, but the overall differences between the two genera were distinct. The Brucella species contained exclusively C16 to C19 acids. Minor amounts of hydroxylated acids (tentatively identified as 3-hydroxy hexadecanoate and 3-hydroxy-octadecanoate) could only be detected after thin layer chromatography and substantial concentration. The Bordetella species, on the other hand, contained a number of both non-hydroxylated and hydroxylated fatty acids of chain-lengths varying from C10 to C19. Intra-generically, no clear distinction between the Brucella species could be detected. The three species of Bordetella, on the other hand, were clearly distinguished by their fatty acid patterns. B. pertussis did not contain cyclopropane fatty acids encountered in significant amounts in the two other species, and B. bronchiseptica was characterized by containing 2-hydroxy-dodecanoate in significant amounts. PMID- 6293246 TI - Binding of Clostridium perfringens enterotoxin to hepatocytes, small intestinal epithelial cells and Vero cells. AB - Clostridium perfringens enterotoxin binds specifically to hepatocytes, Vero cells and rat intestinal epithelial cells, and causes membrane damage visible as "bubble" formation. Fluorescein-labelled enterotoxin also binds specifically to the same cell types. 125I labelled enterotoxin shows one class of binding sites on both Vero and rat intestinal epithelial cells. The number of binding sites decreases along the rat intestine, but the affinity constant is the same in different parts of the intestine. PMID- 6293247 TI - [Neural mechanism of bronchospasmolytic effect of sophocarpine hydrobromide]. PMID- 6293248 TI - [Effects of changrolin and pyracrine phosphate on ATPase of rabbit heart sarcolemma]. PMID- 6293249 TI - [Biochemical mechanism of radioprotection by diethyllipoamide]. PMID- 6293250 TI - Effect of refeeding after starvation on basal and tolbutamide-stimulated insulin secretion and beta-adrenergic receptor function in the regulation of insulin release and lipolysis in obese patients. AB - Serum insulin, blood glucose and plasma free fatty acids (FFA) were determined in 14 subjects with a simple obesity under basal conditions and during the tests with tolbutamide, propranolol and epinephrine before and after fasting of 14 days duration, on restricted diet of 1300 kcal. After refeeding some changes in pancreatic B cells reactivity and an altered metabolic responsiveness to epinephrine and propranolol were found as compared to prefasting values. It may be concluded that after refeeding a further increment of beta-adrenergic function seems to contribute to accelerated lipid mobilisation and partly to increased insulin secretion. PMID- 6293251 TI - [Possible use of the electrophysiological methods for early diagnosis of vibration syndrome]. PMID- 6293253 TI - Nephroblastomatosis. PMID- 6293252 TI - Mechanisms of facilitation and muscarinic or alpha-adrenergic inhibition of acetylcholine and noradrenaline secretion from peripheral nerves. AB - The muscarinic acetylcholine (ACh) receptor from rat brain has been partially purified and characterized. The molecular weight of the smallest subunit of the receptor protein obtained by a chaotropic agent is 30,000 dalton, while the protein found in sodium dodecyl sulphate polyacrylamide gels has a molecular weight of 80,000 dalton. The isoelectric point is in the range 4.5 to 5.1. The low affinity dissociation constant for agonists of the alpha-adrenoceptor from rat and guinea-pig brain, and of the muscarinic ACh receptor from guinea-pig ileum longitudinal muscle-myenteric plexus preparation, is increased 2-3-fold at potassium concentrations of 80-100 mM. The transmitter stores of noradrenergic nerves of guinea-pig vas deferens were labelled by preincubation with 3H noradrenaline (3H-NA), and those of cholinergic nerves of guinea-pig ileum myenteric plexus were labelled by preincubation with 3H-choline. The mechanisms underlying frequency dependent facilitation and presynaptic receptor mediated depression of transmitter secretion were studied. The secretion evoked by electrical stimulation required both invasion of the nerve terminals and activation of the depolarization-secretion coupling in varicosities, while that evoked by high potassium (40-110 mM) was due to direct depolarization varicosities. Blockade of presynaptic alpha-adrenergic and muscarinic autoreceptors by phentolamine and atropine, respectively, caused 4-5-fold greater increase in the secretion of labelled transmitter evoked by electrical than by potassium stimulation from noradrenergic and cholinergic terminals. This suggests that depression of transmitter secretion is exerted mainly by control of the invasion of nerve terminals, and only to a minor extent by control of depolarization-secretion coupling in invaded terminals. Autoinhibition of transmitter secretion involves regulation of calcium availability and is antagonized by facilitation. Endogenous cyclic nucleotides are not critically involved in either facilitation, or alpha-adrenergic or muscarinic autoreceptor mediated control of transmitter secretion in these nerves. PMID- 6293254 TI - Radiation therapy of non-Hodgkin's lymphoma stages I and II. AB - A series of 147 patients, 99 in clinical stage I and 48 in stage II, with localized non-Hodgkin's lymphoma, exclusively given radiation therapy, was retrospectively analysed. Using the Kiel classification, 12 patients (8%) could not be subgrouped with certainty, 63 (43%) were designated as high-graded and 72 (49%) as low-grade malignancies according to the definitions of Gerard-Marchant et coll. (1974). Complete remission was obtained in 93 per cent of the patients in stage I and in 75 per cent in stage II. Most of the failures (68%) were high grade malignancies. Actuarial and relapse-free survival was determined for stage I and II and stratified by microscopy and extranodal disease. All patients initially in stage I had a good prognosis irrespective of the microscopic type; 60 per cent have remained free from disease. In contrast, only 2 patients (4%, all low-grade) in stage II have remained disease-free. Thus, in stage II irradiation cannot be considered the best treatment, especially in patients with high-grade malignancy, in whom chemotherapy may be curable. In stage I, on the other hand, irradiation seems to be curative in the majority of patients irrespective of microscopic type. PMID- 6293255 TI - Correlation between initial salivary flow rate and radiation dose in the production of xerostomia. AB - Dose and initial flow rate are two of several factors that influence the outcome of xerostomia when irradiating head and neck tumors. These two parameters were analysed in 11 irradiated patients where most salivary glands were in the radiation field. Unstimulated whole saliva samples were collected before, during and after the irradiation. The results indicate a statistically significant linear correlation between initial saliva flow rate and the accumulated dose causing irreversible xerostomia. This phenomenon is explained. A diagram is made that could be used (in head and neck tumors where one side could be spared) as a indicator of the maximum dose the salivary glands in the contralateral non involved side can receive before reaching irreversible dryness. PMID- 6293256 TI - Carcinoma of the tongue in Norway and Wisconsin. I. Incidence and prognosis related to sex and age. AB - The records of 503 patients with carcinoma of the tongue diagnosed between 1958 and 1972 were reviewed. The preponderance of tongue carcinoma among men was confirmed both in The Norwegian Radium Hospital (NRH) and the University of Wisconsin Hospitals (UW), but it was relatively more frequent among women in NRH and in UW than in southern Europe. More women had on presentation less advanced tumors at NRH than at UW. The incidence of tongue carcinoma in Norway increased steadily with age for both sexes. The sex ratio did not change in Norway such as in England, Canada and the United States. Tumor of the posterior one-third of the tongue was relatively infrequent in women both in NRH and UW, in agreement with reports from other countries. The length of survival was analysed and no significant sex difference was demonstrated. The younger patients had less advanced tumors and a better prognosis. PMID- 6293257 TI - Irradiation at different times of the day. Morphology and kinetics of the small intestine. AB - Rats were irradiated at different time of the day with sublethal doses on the abdomen only, and qualitative and quantitative morphologic modifications were determined. The experiments seemed to be demonstrate that in the groups irradiated at night and at the end of the light period early injury is not severe whereas in the group irradiated at the end of the dark period repair of the injury seems to be more effective. PMID- 6293258 TI - Amount of peripheral reticulocytes as biologic dosimetry of ionizing radiation. Experiments in the rabbit. AB - The amount of reticulocytes in peripheral blood generally reflect the bone-marrow status of erythropoiesis and may therefore be useful in evaluating radiation injury. The blood reticulocytes in rabbits exposed to various doses of ionizing radiation were examined using an apparatus capable of microfluorometric quantification of reticulocytes as well as the conventional technique of reticulocyte count. A dose-dependent decrease of reticulocytes was observed. The possibilities of further improvements of the technique using flow cytometry and its application for screening human population are discussed. PMID- 6293259 TI - Carcinoma of the ovary in stage III. Effects of postoperative chemotherapy, radiation therapy and repeat laparotomy. AB - A prospective randomized trial was carried out in 153 patients with stage III malignant epithelial tumours for comparing the effects of irradiation or combination of chemotherapy and irradiation on prognosis and operability. No significant differences between these two treatment modalities were found. Of the patients primarily considered inoperable were 41 per cent operated upon after preoperative treatment. Minimum residual disease (0 to less than or equal to 2 cm) occurred in 38 per cent of the primarily operated and in 31 per cent of those operated upon after preoperative treatment. Preoperative irradiation in primarily inoperable patients enabled more effective surgical measures at relaparotomy. The size of the residual tumour after surgery and the tumor grade influenced the survival. PMID- 6293260 TI - Radiation therapy in primary carcinoma of the vagina. AB - The experiences with irradiation of 25 patients with primary carcinoma of the vagina are presented. The 3-year survival was 36 per cent despite the fact that most of the cases were advanced. Further improvements in survival may probably be achieved with extension of the radium implantation to paravaginal tissues. Choice and technique of treatment must be individualized. Surgery is limited to the more advanced cases with involvement of the bladder and rectum. PMID- 6293261 TI - Role of radiation therapy in the management of primary carcinoma of the vagina. AB - A retrospective analysis of 88 patients with microscopically proven primary carcinoma of the vagina is presented. All patients were treated between 1957 and 1975 and have been followed for a minimum of 5 years. The technique of treatment, external, intracavitary and interstitial irradiation has been highly integrated and individualized. Several important features could be drawn from the current data: (1) The necessity for individualized radiation treatment to achieve the success, (2) the necessity to subdivide stage II into IIA and IIB is clearly shown by the 5-year survival data, (3) a relatively low complication rate from the use of high dose combined radiation modalities in higher stage is demonstrated. PMID- 6293262 TI - Intercomparison of Nordic therapy level exposure secondary standards for 60Co gamma radiation. AB - An intercomparison of therapy level exposure secondary standards from the Nordic countries was made at 60Co gamma radiation at the secondary standard dosimetry laboratory in Helsinki . One standard seemed to be in error by about one per cent and another was found to be unreliable. From the analysis it was concluded that inter-comparisons of secondary standards can be made with an overall uncertainty of about 0.2 per cent. This uncertainty is of the same size as the difference between primary standards, which may therefore play a role in the evaluation of differences between secondary exposure standards. PMID- 6293263 TI - Carcinoma of the tongue in Norway and Wisconsin. II. Influence of site and clinical stage on local control of the tumor. AB - The local control rates in patients with squamous cell carcinoma of the tongue, treated between 1958 and 1972, were determined by retrospective analysis of the records of 339 patients at the Norwegian Radium Hospital and of 164 patients at the University of Wisconsin Hospitals. The correlation between the clinical stage and the local control rate was good in Norway, less striking in Wisconsin. As the tumor grew in size it generally became increasingly difficult to control. Irradiation and surgery appeared relatively satisfactory in patients with tumors in early stages, but were often inadequate in the advanced tumors. The differences observed between Norway and Wisconsin were statistically significant only for the tumors of the base of the tongue and T2N0, T3N0 and TxN1 tumors of the anterior two-thirds of the tongue. PMID- 6293264 TI - Influence of adjuvant chemotherapy on the blood lymphocyte population in operable breast carcinoma. Comparison between two types of treatments. AB - The influence on the blood lymphocyte population of two types of postoperative adjuvant chemotherapy regimes given to patients with large breast tumors or involved axillary lymph nodes have been examined. Cyclic treatment with a combination of chlorambucil, methotrexate and 5-fluorouracil was more myelotoxic and required more extensive dose reductions than treatment with cyclophosphamide, methotrexate and 5-fluorouracil with the dosage used. Both treatments reduced the size of the blood lymphocyte population and changed its cellular composition, as defined by rosette tests, to approximately the same extent. Response of the lymphocytes to specific and non-specific mitogens were also affected to approximately the same extent. A comparison of the clinical value between the two treatments has not yet been performed. PMID- 6293265 TI - Influence of treatment and tumor grade on the prognosis of stage II carcinoma of the endometrium. AB - A retrospective analysis of 53 patients with stage II endometrial carcinoma treated between 1963 and 1975 was undertaken to evaluate the efficacy of treatment methods and the effect of tumor grade on survival and failure. Thirty eight patients were irradiated preoperatively followed by total abdominal hysterectomy and bilateral salpingo-oophorectomy, 15 were followed for at least 5 years with no lost to follow-up. An overall 5-year disease-free survival of 60.6 per cent was obtained. Further analysis of the data showed that tumor grade and treatment method were important factors influencing the prognosis. The data support the improved survival rate when irradiation is followed by hysterectomy. Patients with a low grade tumor (G1) given preoperative irradiation have a better survival than those with high grade tumor (G3) treated by the same method. Failure rate is also dependent on method of treatment and tumor grade. The results are in agreement with several data in the literature that combination of the irradiation and surgery should be the treatment of choice for stage II endometrial carcinoma. Tumor grade is also an important factor and must be evaluated for the extension of the disease before the proper management. For the high grade tumor, para-aortic node sampling might be important for the irradiation and should warrant further investigation. PMID- 6293266 TI - Technique for microwave-induced hyperthermia in superficial human tumours. AB - In order to induce local hyperthermia in superficial tumours a computer system using a 2,450 MHz microwave generator connected to a circular (diameter 90 mm) direct contact applicator was constructed based on invasive temperature control. Small thermistor probes with a diameter of 0.6 mm are inserted in the tumour and surrounding tissues. Eight thermistors can be used simultaneously and one of them is chosen as the "master'. The automatic control system used a pulsed irradiation technique to avoid the problem with interactions between the metallic wires of the temperature probe and the electromagnetic field. With the system it was possible to control the temperature at the master thermistor within +/- 0.5 degrees C from the preset value. PMID- 6293267 TI - Microwave-induced hyperthermia and ionizing radiation. Preliminary clinical results. AB - The combination of microwave-induced (2450 MHz) hyperthermia and ionizing radiation was used in 7 patients with superficial malignant tumours, which were considered refractory to other therapy. A newly developed heating system was used, allowing for a maintained temperature at the master probe of 42.5 degrees C +/- 0.5 degrees C during 45 min, but temperature measurements at multiple sites showed a marked variation. This preliminary series indicates that the combination of hyperthermia and ionizing radiation may be useful, the response rate (complete or partial) being 8 of 8 evaluable lesions. Even previously heavily irradiated sites responded. Technical improvements are highly needed to allow for controlled heating of any tissue volume. PMID- 6293268 TI - Growth kinetics of Bp8 mouse ascites sarcoma after single dose whole body irradiation. I. Analysis of the relative and total numbers of cells in various parts of the cell cycle. AB - The effects of irradiation on the growth of the Bp8 mouse ascites sarcoma were analysed following doses of 1.75, 2.5, 5.0 and 8.0 Gy. From the total number of cells and the percentage of cells in G1, S-phase, G2 + M as measured by flow cytofluorometric DNA analysis and the mitotic index the total number of cells in the various parts of the cell cycle was estimated. After an initial delay in the increase in the cell numbers during the period of rapid growth the total number of cells shows a dose dependent decrease at the plateau stage of the ascites growth. This dose relationship is characterized by a shoulder type of curve with a D0 of about 6.5 Gy and Dq of about 2 Gy. This decrease in the total number of cells is caused by a decreasing number of G1, S-phase and mitotic cells while G2 cells generally remain at unchanged levels. This behaviour of the G2 cells reflects the preference of long-lasting blocking events in G2 of the cell cycle but may also indicate specific regulating processes linked to this type of cell. The ratio between the number of mitotic and G2 cells also decreases in a dose dependent way and is a sensitive indicator for irradiation effects even below the lowest dose (1.75 Gy) used in the present experiments. PMID- 6293269 TI - Permeability of the blood-brain barrier in the rat after local proton irradiation. AB - Rats were irradiated laterally through the brain with 200 MeV protons. The beam was of circular cross-section with a diameter of 5 or 7 mm. The doses were 50, 70, 100 and 150 Gy. After irradiation the rats were examined several times by use of injected 99Tcm-pertechnetate. The uptake of the substance increased to a maximum after 20 to 30 days and then decreased to a normal level. Differences in maximum uptake with respect to dose were significant only for the smaller beam diameter. PMID- 6293270 TI - Modifications in the brush border enzymes of the small intestine after irradiation at different times of the day. AB - The behaviour of the brush border enzyme activity of the intestinal epithelium after the same sublethal radiation dose to the abdomen at different times of the day was investigated. Three previously observed post-irradiation phases (initial increase of activity, reduction and the return to control values) were confirmed, although with some differences. A later return to normal of lactase was also confirmed. The same dose produced different behaviour of the enzyme activities both during the initial and the recovery phase, depending on the time of the day when irradiation was performed, i.e. on the functional condition of the epithelial cells. PMID- 6293271 TI - Comparison of neutron therapy beams produced by 50 MeV deuterons and 65 MeV protons on beryllium. AB - Neutron beams produced by bombarding a 10 cm thick beryllium target with 50 MeV deuterons have been used at Louvain-la-Neuve since nearly 4 years for routine therapeutic applications. At the end of 1981 they were replaced by neutron beams produced by 65 MeV protons on beryllium, mainly in order to improve the beam penetration in tissues. However, produced of neutrons from the p leads to Be reaction implies some disadvantages, mainly a lower dose rate and a higher activation level. In order to solve the problem a new target configuration was designed, consisting of a remote handled system which permits the use of 2 different target assemblies. The irradiated target is automatically removed immediately after the irradiation which greatly protects against exposure to the staff. The dosimetric characteristics of the d(50)-Be and p(65)-Be neutrons beams are compared. PMID- 6293272 TI - Sonomotor reflex as a descending control mechanism in the auditory system in man. PMID- 6293273 TI - Isolated thyrotropin deficiency in a man with narcoleptic attacks. AB - A patient with narcoleptic attacks and isolated thyrotropin deficiency is presented. Substitution with l-thyroxine did not relieve his narcoleptic attacks. Deficiency, in the central nervous system, of the ergotropic substance thyrotropin-releasing hormone (TRH) might have caused the thyrotropin deficiency and contributed to the narcoleptic attacks. However, prolonged oral medication with TRH, in doses of 120 or 240 g daily, failed to attain any significant relief. Although this does Not exclude TRH unresponsiveness due to absent or defective TRH receptors, the true cause of the disease remains to be elucidated. PMID- 6293274 TI - The effect of Metamucil on postprandial blood glucose and plasma gastric inhibitory peptide in insulin-dependent diabetics. PMID- 6293275 TI - [Biological effect of D,L-ureidosuccinic acid dihydrazide on Pseudomonas pseudomallei and Salmonella typhimurium]. PMID- 6293276 TI - [Eruptive hidradenoma]. PMID- 6293277 TI - The activity of substituted benzamides in biochemical models of dopamine receptors. PMID- 6293278 TI - High-performance liquid chromatography: applications in a children's hospital. PMID- 6293279 TI - Chromatography of vitamin D3 and metabolites. PMID- 6293280 TI - A method for studying the dynamics of the primary migration of human lymphocytes using Indium-iii oxine cell labelling. PMID- 6293281 TI - Study of beta adrenoceptors on subpopulations of thymocytes separated on ficoll hypaque density gradients. PMID- 6293282 TI - The thymus-hypophysis interaction in the developing chick embryo: thymic epithelial cells in hypophysectomized embryos. PMID- 6293283 TI - Human sessile B lymphocyte marker defined by a monoclonal antibody. PMID- 6293284 TI - Impaired host resistance against local JMV Marek's disease tumors after thymectomy and after bursectomy. PMID- 6293285 TI - Helper T cells specific for protein antigens: role of self major histocompatibility complex and immunoglobulin gene products. PMID- 6293286 TI - Limbic innervation of the striatum. PMID- 6293288 TI - Penetration of viral genetic material into host cell. PMID- 6293287 TI - Participation of prostaglandins in the control of renin release. PMID- 6293289 TI - Mechanisms of viral tumorigenesis. PMID- 6293290 TI - Studies of Japanese encephalitis in China. PMID- 6293291 TI - Pyridine nucleotide metabolism in the erythrocyte of South African blacks with primary hepatoma. AB - Erthrocytes from African blacks with primary hepatoma were incubated with physiological amounts (1.64 microM) of nicotinamide-14C (NM-14C) and it was found that these erythrocytes could synthesize NAD from NM. After 3-hr incubation with NM-14C, a large percentage of the 14C was found in NMN, nicotinamide riboside (NR) and NAD, but was undetectable in nicotinic acid nucleotides (NAMN and NAAD). This suggested that the NAD synthesized from NM was not through the Preiss Handler pathway. After 6-plus hr incubation, the 14C found in NAMN and NAAD suggested the NAD synthesized was being broken down and reutilized through Preiss Handler pathway for synthesis of NAD. This reutilization pathway was confirmed by incubating nicotinic acid-14C (NA-14C) with erythrocytes. Apparently the metabolites from the breakdown of NAD were deaminated. The metabolism of NM-14C was slower than NA-14C. However, after 24 hr incubation with NM-14C, 72.26% of 14C was found in NAD. A high percentage of 14C in NR at the initial incubation and a later drop suggested that NR was another intermediate in the pathway. PMID- 6293293 TI - Dynamic computed tomography of hepatocellular carcinoma. AB - Nineteen patients with clinically proven hepatocellular carcinoma were studied by dynamic computed tomographic (CT) scanning. Ten consecutive 3 sec scans were performed within 30 sec, providing uninterrupted data collection. Dynamic CT scanning was useful in recognizing tumor vascularity, multiple small tumors, isodense tumors on conventional CT scans, the capsule of an encapsulated hepatocellular carcinoma, arteriovenous shunts, and vascular pools. Time-density curves were useful in evaluating the hemodynamics of the tumors; these could be classified into three types according to differences in their enhancement patterns. PMID- 6293292 TI - [Protection of essential fatty acids by vitamin E]. AB - The protective role of vitamin E against free-radical-mediated oxidations is discussed. In spite of the presence of vitamin E in cell membranes, as structural complex with polyunsaturated fatty acids (PUFA) of phospholipids, the question arises whether high PUFA containing diets, producing high deposition of PUFA in the tissues, can nevertheless lead to peroxidations in the body. It has been suggested that large amounts of dietary PUFA increase the requirement for vitamin E and deplete its tissue stores, particularly when PUFA are discontinued in the diet, also because of their longer half-life time than tocopherols. However, in physiological conditions, linoleic acid up to 10% of caloric intake seems to have no effects on vitamin E requirement. In contrast, in essential fatty acid (EFA) deficient animals, also the addition of small amounts of dietary EFA, by resulting in a proportional increase in PUFA content of membrane structural lipids, is associated with an increased need for vitamin E. This becomes particularly important in the case of dietary fish oils or other poorly protected fats. PMID- 6293294 TI - Periodontal pocket elimination using Durapatite ceramic implant. PMID- 6293295 TI - Acute effects of the calcium antagonist, nifedipine, on blood pressure, pulse rate, and the renin-angiotensin-aldosterone system in patients with essential hypertension. AB - Ten milligrams nifedipine was administered orally to young and old persons with or without hypertension, and the acute effects of nifedipine on the renin angiotensin-aldosterone system were studied one half to 3 hours later. Nifedipine reduced blood pressure and increased pulse rate in young and old persons with or without hypertension. Simultaneously, nifedipine produced a significant increase of plasma renin activity in young persons with or without hypertension but failed to do so in old persons with or without hypertension. As a result, angiotensin I and II increased significantly in young persons but not in old persons. Hydralazine elevated aldosterone concentration by stimulating the renin angiotensin system but nifedipine failed to do so despite its effect on the renin angiotensin system in young individuals. Since calcium is required to secrete aldosterone, it is suggested that nifedipine blocked aldosterone secretion by the agent's calcium antagonizing action. PMID- 6293297 TI - Effects of dietary fiber on intestinal ion fluxes in rats. AB - The effect of short-term fiber ingestion on jejunal ion fluxes was evaluated in rats using a standard Ussing chamber technique. Ingestion of cellulose and pectin decreased the mucosal to serosal fluxes of both Na and Cl but did not significantly alter serosal to mucosal fluxes; net fluxes of both Na and Cl were significantly lower in the group supplemented with dietary fiber as compared to those animals fed a fiber-free diet. Both potential difference and short-circuit current were higher in the fiber-free group than in the group supplemented with dietary fiber; tissue conductance, however, was unaffected by fiber ingestion. The residual flux of all three groups was virtually identical suggesting that electrical alterations observed after cellulose and pectin ingestion are not the result of ion fluxes other than Na and Cl. These data, coupled with previous observations that short-term fiber supplementation impairs glucose and leucine absorption, suggest that fiber ingestion alters the intestinal membrane, specifically sites of active transport. PMID- 6293296 TI - Effect of processing on digestibility and the blood glucose response: a study of lentils. AB - To test the effect of processing on digestibility and the glycemic response to a leguminous seed, a group of eight healthy volunteers took a series of breakfast test meals containing either lentils which had been processed in four different ways or the same amount of carbohydrate as white bread. Lentils, boiled for 20 min, resulted in a flattened blood glucose response by comparison with bread. This was unaltered by blending the lentils to a paste or boiling them for an additional 40 minutes. However the blood glucose response was significantly enhanced by drying the boiled blended lentils for 12 h at 250 degrees F. In vitro digestion with human saliva showed the rate of sugars released from the food related positively to the blood glucose rise. Breath hydrogen studies indicated that carbohydrate malabsorption was too small to account for differences in the blood glucose response. These results emphasize the importance of processing in determining digestibility and hence the glycemic response to a food. PMID- 6293298 TI - Human linolenic acid deficiency. PMID- 6293299 TI - Pathologic predictors of recurrence in stage 1 (TINOMO) breast cancer. AB - A group of 122 consecutively treated pathologic Stage 1 (TINOMO) patients with cancer of the breast were studied to define histopathologic predictors of recurrence. Lymphatic invasion was the most significant predictor of recurrence; recurrence was present in 32% (8/25) of patients who had lymphatic invasion and in 10.3% (10/97) of patients who did not (P = 0.006). Histologic type was also predictive of recurrent disease. Eighteen per cent (18/101) of patients with invasive ductal or lobular carcinoma developed recurrent disease, while none of the group of 21 patients with medullary carcinoma, tubular carcinoma, colloid carcinoma, Paget's disease, and intraductal carcinoma with minimal invasion suffered a recurrence (P = 0.036). Vascular invasion, grade of malignancy, cellularity, presence or absence of circumscription, cellular infiltrate, fibroblastic response, neural invasion, and necrosis were not significant predictors of recurrence. Multiple logistic regression analysis of patients with invasive ductal or lobular carcinoma confirmed the results for individual factors, that only patients with lymphatic invasion were at higher risk of recurrence. PMID- 6293300 TI - Hormone receptor levels and aspiration cytology in cancer of the breast. AB - The cytopathologic findings of 196 cases of cancer of the breast were correlated with the hormonal receptor levels. The strict cytologic criteria that allow a positive diagnosis to be used as bases for definitive treatment include adequate cellularity, lack of cohesion, and nuclear abnormalities, including increased nuclear size. On this basis, 58% of the carcinomas of the breast, including 62% of the 157 infiltrating duct carcinomas but none of 12 infiltrating lobular carcinomas, could be diagnosed by aspiration smears alone. Seventy-four per cent of the carcinomas were estrogen-receptor-positive. No correlation could be demonstrated between the cytologic pattern and diagnosis and the level of estrogen, progesterone, or androgen receptors, alone or in any combination. PMID- 6293301 TI - Congenital cytomegalovirus infection in a pair of dizygotic twins. PMID- 6293302 TI - Coxsackievirus B3 producing fatal meningoencephalitis in a patient with X-linked agammaglobulinemia. PMID- 6293303 TI - Antibody to hepatitis A and hemodialysis. AB - Antibody to hepatitis A virus (anti-HAV) was surveyed in 469 patients from 20 of 31 Michigan hemodialysis units, during spring 1978. The mean point prevalence of anti-HAV was 59.5% and within the 20 individual units ranged from zero to 100%. For the entire survey population, the point prevalence of anti-HAV was significantly greater with increasing age, among blacks, and in individuals with hypertension as their underlying renal disease. Anti-HAV was independent of sex, duration of dialysis, or the presence of either hepatitis B surface antigen or its associated antibody. Within individual units, anti-HAV prevalence was associated with a higher mean patient age but not with dialysis unit size, mean duration of dialysis therapy, race, or prevalence of hepatitis B markers among patients or staff. These data support reports that transmission of the hepatitis A virus is neither associated with hemodialysis therapy nor routinely spread by parenteral mechanisms as observed in hepatitis B virus transmission. PMID- 6293304 TI - Transmission of experimental rhinovirus infection by contaminated surfaces. AB - Transfer of experimental rhinovirus infection by an intermediary environmental surface was examined in healthy young adults, in four studies done in 1980--1981, by having recipients handle surfaces previously contaminated by infected donors. Recipients touched their nasal and conjunctival mucosa after touching the surfaces. Five (50%) of 10 recipients developed infection after exposure to virus contaminated coffee cup handles and nine (56%) of 16 became infected after exposure to contaminated plastic tiles. Spraying of contaminated tiles with a commercially available phenol/alcohol disinfectant reduced (p = 0.003) the rate of recovery of virus from the tiles from 42% (20/47) to 8% (2/26). Similarly, the rate of detection of virus on fingers touching the tiles was reduced (p = 0.001) from 61% (28/46) with unsprayed tiles to 21% (11/53) with sprayed tiles. Fifty six per cent (9/16) of the recipients exposed on three consecutive days to untreated tiles became infected while 35% (7/20) touching only sprayed tiles became infected with rhinovirus (p = 0.3). These studies indicate that experimental rhinovirus colds can be spread by way of contaminated environmental surfaces and suggest that disinfectant treatment of such surfaces may reduce risk of viral transmission by this route. PMID- 6293305 TI - An outbreak of Norwalk gastroenteritis associated with swimming in a pool and secondary person-to-person transmission. AB - In June 1977 an outbreak of acute gastroenteritis affected 103 students and teachers at an elementary school in Ohio. The illness typically lasted 24 hours or less and was characterized by vomiting (86%) and cramping (70%), but more than half of the persons involved also reported having nausea, diarrhea, and headache. Similar illness frequently followed in household members (29%) of families with primary cases. Investigation revealed that 70% of the children and teachers who swam in a pool at an all day outing June 1 (4 classrooms) and 55% of those who swam during a similar outing June 2 (2 classrooms) had the onset of acute illness from 12--48 hours later. None of the children who attended the outings but did not swim had a similar illness. The evidence suggested that the primary outbreak was caused by contaminated water in the pool and that person-to-person spread of illness followed. Results of a microbiologic study of pool water were negative for bacterial and viral pathogens. Throat washings, stool specimens, and paired blood samples studied for evidence of pathogens were negative initially, but subsequent serologic studies suggested that infection by Norwalk virus was the cause of the outbreak. The pool chlorinator which was inadvertently unconnected at the time of the school visits was reconnected and an underground leak in the water supply pipes was corrected. No more cases were reported after the pool was drained, cleaned, and reopened. PMID- 6293306 TI - An outbreak of acute nonbacterial gastroenteritis in a nursing home. Demonstration of person-to-person transmission by temporal clustering of cases. AB - An outbreak of acute nonbacterial gastroenteritis occurred among residents and staff in a nursing home in Baltimore, Maryland, in December 1980. A total of 101 residents and 69 staff members were surveyed by questionnaire. The attack rate (defined as acute onset of vomiting or two or more loose stools per 24 hours) was 46% in each group. Illness was brief and mild; no patients were hospitalized, and there were no deaths. Person-to-person transmission was documented by temporal clustering of cases (the demonstration of higher rate of illness among residents exposed to an ill roommate one or two days earlier than among those not similarly exposed; relative risk = 3.74), by a higher rate of illness among employees having daily contact with residents than among those without such contact (57% vs. 17%, p less than 0.01), and by secondary transmission to household contacts of ill employees (secondary attack rate = 33%). Three of 11 serum pairs from patients demonstrated a fourfold increase in antibody titer to the Norwalk virus between acute- and convalescent-phase specimens. The analysis of temporal clustering of cases was particularly useful in documenting person-to-person transmission in this outbreak and might be used for this purpose in other outbreaks caused by Norwalk or Norwalk-like viruses, as well as in outbreaks associated with other infectious organisms. PMID- 6293308 TI - The incidence and outcome of asymptomatic herpes simplex genitalis in an obstetric population. AB - In a prospective study of an obstetric population, 28 of 488 (4%) routine asymptomatic patients had positive cultures for herpes simplex virus from their genital tract. Cesarean section was carried out if the virus was present in the genital tract at the time of delivery. In one instance, a culture which had reverted to negative became positive again. No cases of neonatal herpes were reported in the time of the study. As yet, the infectivity of these subclinical cases is unclear, and the incidence of neonatal herpes is not as high as would be expected from the study. Further studies need to be carried out to evaluate the exact risk of the asymptomatic herpesvirus shedder to the infant at birth. PMID- 6293307 TI - Herpes zoster in patients with carcinoma of the lung. AB - Herpes zoster was observed in only four of 250 (1.6 percent) patients with small cell carcinoma of the lung, who were treated in a prospective, combined modality therapy trial. Induction chemotherapy in this study consisted of six courses of cyclophosphamide, doxorubicin, and vincristine (CAV), followed by intrathoracic and cranial irradiation. Those with extensive disease also received single doses of upper half-body irradiation. Patients did not receive maintenance chemotherapy (CAV2 protocol). This contrasted with our previous study (CAV1 protocol), which consisted of three courses of the same induction chemotherapy, the same intrathoracic irradiation, but with one year of oral maintenance chemotherapy. During the CAV1 regimen, we observed that herpes zoster developed in 13 of 161 (8.1 percent) patients in association with their therapy. A retrospective analysis of 6,576 patients with lung cancer revealed that herpes zoster developed in 58 (0.9 percent). This complication developed in 10 of 622 (1.6 percent) patients with small cell carcinoma of the lung, as compared to 48 of 5,954 (0.8 percent) patients with non-small cell carcinoma of the lung. The risk of development of herpes zoster in the CAV1 group was significantly greater than the historical group (p = 0.007) and was also greater than the CAV2 group (p = 0.031). However, there was no significant difference between the historical group and the CAV2 group. Attempts to explain the differences in the rate of herpes zoster in our three studies and those in the literature suggest that the duration of therapy, the type of chemotherapy used, and the improving survival rate may be important contributing factors to this complication in patients aggressively treated for small cell carcinoma of the lung. The literature and our own studies suggest that procarbazine is the most likely chemotherapeutic agent predisposing to this complication. PMID- 6293309 TI - Adenoid cystic carcinoma. PMID- 6293310 TI - A possible mechanism of inotropic action of prolactin on rat heart. AB - Prolactin possesses positive inotropic actions in isolated heart preparations although the mechanism of this influence is not understood. Our study was designed to investigate the mechanism of this effect on the rat heart. Prolactin (50 ng/ml) produced a time-dependent increase (60%) in contractile force that reached maximum after 30 min and remained steady for a further 30 min. A similar time-dependent phenomenon was seen with 200 ng/ml prolactin although the maximum inotropic effect was reduced. Indomethacin (30 micrograms/ml) significantly reduced the inotropic effect of both prolactin concentrations although the effect of the hormone was not related to the release of 6-keto-PGF1 alpha, the prostacyclin metabolite. Propranolol (1-20 micrograms/ml) significantly reduced the positive inotropic effect of prolactin. Prolactin however had no influence on myocardial adenylate cyclase activity. Hearts that were removed from animals pretreated with 1.25 or 2.50 mg/kg reserpine did not respond to prolactin administration. It is suggested that the inotropic influence of prolactin is mediated by endogenous catecholamine liberation. PMID- 6293311 TI - Effect of hypertonic intracarotid infusions on plasma vasopressin concentration. AB - The effect of short-term bilateral intracarotid infusions of hypertonic saline on plasma vasopressin concentration (pAVP) was evaluated in five dogs. Intracarotid infusion of saline at 90 mumol . kg-1 . min-1 . artery-1 significantly (P less than 0.05) increased jugular vein osmolality (pOsm) and sodium concentration (pNa+) within 2 min. Saphenous vein pOsm was not altered during the 6 min of infusion, whereas pNa+ was increased (P less than 0.05) from 0.8 +/- 0.1 to 2.3 +/- 0.3 pg/ml. Subsequent experiments using hypertonic saline infusions of 90 and 180 mumol . kg-1 . min-1 administered intracarotidly and intravenously for 6 min were performed. Intracarotid isotonic infusions and intravenous hypertonic infusions did not significantly alter pAVP. Hypertonic intracarotid saline increased jugular vein pOsm and pNa+ in a dose-related fashion, whereas saphenous vein pOsm and pNa+ were not significantly changed after 6 min of infusion. Plasma vasopressin, compared with the isotonic intracarotid infusion (1.5 +/- 0.3 pg/ml), was increased (P less than 0.05) after hypertonic saline to 3.2 +/- 0.6 and 4.8 +/- 0.2 pg/ml for the 90 and 180 mumol . kg-1 . min-1 infusions, respectively. The cerebral osmolality indicated by jugular vein pOsm was therefore increased in the absence of changes in systemic pOsm during intracarotid hypertonic infusions. The increase in pAVP in response to these changes in pOsm supports the presence of central osmoreceptors regulating vasopressin release in the area of distribution of the common carotid arteries. PMID- 6293312 TI - Interpretation and use of electrical equivalent circuits in studies of epithelial tissues. AB - Whereas transepithelial and intracellular voltages continue to be measured in renal and other epithelial tissues, the origins of these voltages, especially in renal epithelia, remain obscure. Because epithelial tissues have multiple transcellular and extracellular routes of ion transport, it is convenient to model them with electrical equivalent circuits and, in this way, attempt to understand the relative importance of and relationships between the parallel series arrangements of the membranes and barriers involved. The interpretation of the equivalent electromotive forces and resistances can be complicated, however, by virtue of nonlinear current-voltage relationships of ionic channels. Thus, for ion transport pathways displaying nonlinear I-V relationships, it is important to distinguish between chord and slope formalisms in the use and interpretation of electrophysiological data. For ions like Na that are generally not at electrochemical equilibrium, the Thevenin electromotive force (emf) of the slope formalism is not synonymous with the Nernst equilibrium potential of the chord formalism nor are the slope and chord conductances equal or constant at all voltages. Thus, it is mandatory that the empirical data be calculated and interpreted in a way consistent with the formalism adopted. The existence of nonlinear behavior, characterized by either Goldman or other types of rectification, exacerbates determination of relative ionic permeabilities, fractional resistances, transference numbers, and other electrophysiological parameters for simple membranes and especially for epithelia. It is argued that the use and interpretation of electrical equivalent circuits of epithelia are not arbitrary but must take into account nonlinearities of the ionic current-voltage relationships and concentration and voltage dependencies of the emfs and conductances. PMID- 6293314 TI - Renal Na+-K+-ATPase in renin release. AB - The effects of ouabain and furosemide on renin secretion, renal function, and renal Na+-K+-ATPase were investigated in anesthetized dogs. Furosemide (2 mg/kg) induced significant diuresis, natriuresis, an increase in renal blood flow (RBF), and a fivefold increase in renin secretory rate (RSR), but no changes in glomerular filtration rate (GFR). Infusion of ouabain (1 microgram . kg-1 . min 1) into one renal artery during furosemide diuresis increased fractional sodium excretion from 22 +/- 2 to 30 +/- 3% from the ipsilateral kidney but did not change urine flow, RBF, or GFR, whereas RSR fell to control values (698 +/- 203 to 137 +/- 43). When ouabain preceded furosemide, the rise in RBF and RSR induced by furosemide was abolished but sodium excretion increased. Ouabain infused in vivo inhibited Na+-K+-ATPase in microsomal fractions from cortex (34%) and medulla (27%) as compared with control. Neither saline nor furosemide exerted any effect on Na+-K+-ATPase. Moreover, the effect of ouabain alone on Na+-K+-ATPase was not different from that of ouabain plus furosemide. No changes in Mg2+-ATPase were detected in any of the experiments. These results indicate that inhibition of renal Na+-K+-ATPase abolishes furosemide-induced renin secretion despite potentiation of the natriuretic effect of the diuretic. It is apparent that the level of activity of Na+-K+-ATPase is of prime importance for renin secretion. In addition, ouabain may act directly on the juxtaglomerular cells to inhibit renin secretion. PMID- 6293313 TI - Effects of metabolic acidosis on PTH and 1,25(OH)2D3 response to low calcium diet. AB - To study the effects of chronic metabolic acidosis on the metabolism of 1,25 dihydroxyvitamin D3 [1,25(OH)2D3] rats were given either a low calcium diet (LCD) (0.002% calcium) or chow (1.2% calcium); ammonium chloride (NH4Cl) was added (1 or 1.5%) to the drinking water of some rats eating LCD or chow while others served as nonacidotic controls. LCD increased circulating 1,25(OH)2D3 levels from 46 +/- 14 to 204 +/- 24 pg/ml (P less than 0.001) in the absence of NH4Cl; 1.5% NH4Cl prevented the increase in 1,25(OH)2D3 (25 +/- 6 vs. 27 +/- 8 pg/ml (P, NS) but 1% NH4Cl did not (50 +/- 12 vs. 161 +/- 23 pg/ml; P less than 0.001). Acidosis suppressed neither serum immunoreactive parathyroid hormone (PTH) nor urine cAMP response to LCD. Although total serum calcium and phosphorus showed no regular changes with NH4Cl, acidosis raised blood ionized calcium in rats fed either chow or LCD, and serum 1,25(OH)2D3 levels were inversely correlated with ionized calcium (r = 0.714; P less than 0.001) during LCD. Chronic NH4Cl acidosis prevented serum 1,25(OH)2D3 from rising during LCD, independent of changes in PTH secretion, cAMP generation, or serum phosphorus. The absence of a 1,25(OH)2D3 response may be due to increased ionized calcium produced by acidosis. PMID- 6293315 TI - Mucosal gastrin receptor. VIII. Sex-related differences in binding. AB - The purpose of the current study was to determine whether gastrin binding differed between the sexes and, if so, to ascertain the factors responsible. Male and female rats were killed at various times from 15 to 60 days after birth. Binding capacities in male and female rats were equal until puberty. By day 40 there were significantly more receptors present in males than females. Females had 2.4 fmol receptor/mg prot by day 40, and this value remained constant into adulthood. In male rats the number of receptors continued to increase to 4.0 fmol/mg prot at day 60. Castration did not significantly decrease the number of receptors in male rats. Ovariectomy, however, increased the number of receptors in female rats to a value not significantly different from male controls. Ovariectomized females consumed more food and had higher serum gastrin levels. If ovariectomized rats were pair fed to the level of intact females, the number of receptors and serum gastrin levels decreased over those present in ovariectomized rats fed ad libitum but still remained significantly higher than those in intact females. Adrenalectomy of ovariectomized females caused a further increase in receptor number. PMID- 6293317 TI - Inhibition of acid secretion in isolated gastric glands by substituted benzimidazoles. AB - A new class of gastric acid inhibitors, substituted benzimidazoles (H 83/69 and H 149/94), have been tested in an isolated rabbit gastric gland preparation. Acid formation in the glands was stimulated by histamine, dibutyryl cAMP (DBcAMP), and high extracellular K+ concentrations, and the glandular secretory response was measured by changes in oxygen consumption and in accumulation of the weak base [14C]aminopyrine (AP). The substituted benzimidazoles inhibited AP accumulation induced by all stimulants in a dose-dependent noncompetitive manner. In contrast, cimetidine only inhibited histamine-induced AP accumulation. Basal AP accumulation, not affected by cimetidine, was also inhibited by the substituted benzimidazoles, as was the increase in glandular oxygen consumption produced by the addition of histamine and DBcAMP. Basal oxygen consumption was inhibited by about 15%. The substituted benzimidazoles, like AP, are weak bases and were also found to accumulate in the glands. Semiquantitative morphological studies of glands stimulated by histamine plus theophylline did not show any change in the enlarged secretory surface area after stimulation in the presence of inhibitory concentrations of H 149/94 (10(-4) M). The results suggest that substituted benzimidazoles have a mechanism of action different from that of H2-receptor antagonists and indicate a very distal site of action in the events leading to acid formation. PMID- 6293316 TI - Cholecystokinin-induced contraction of dispersed smooth muscle cells. AB - Isolated smooth muscle cells were prepared from the fundus of guinea pig stomach by incubation with collagenase. Incubating the cells with the C-terminal octapeptide of cholecystokinin induced contraction, which was measured by micrometry and expressed as percent decrease in mean cell length. Cholecystokinin induced contraction was maximal within 30 s and reduced cell length by approximately 37%. The threshold concentration of cholecystokinin was 0.1 pM, and the maximally effective concentration was 0.3 nM. Contraction caused by cholecystokinin could be inhibited by proglumide and by glucagon. Inhibition by proglumide was competitive and resulted in a parallel rightward shift of the cholecystokinin dose-response curve. In contrast, inhibition by glucagon was noncompetitive and resulted in a reduction in the efficacy of cholecystokinin without a change in its potency. Furthermore, proglumide-induced inhibition was specific for cholecystokinin, whereas glucagon-induced inhibition of contraction was nonspecific and reduced the contraction caused by carbamylcholine and the calcium ionophore A23187. PMID- 6293319 TI - Are we for mental health as well as against mental illness? The significance for psychiatry of a global mental health coalition. PMID- 6293318 TI - Ouabain binding and potassium relaxation in aortas from renal hypertensive rabbits. AB - This study was designed to characterize the electrogenic sodium pump in vascular smooth muscle from one-kidney one-clip (1K-1C), renal hypertensive rabbits. Two measures of the electrogenic pump were used: 1) [3H]ouabain binding and 2) potassium-induced relaxation. The binding study indicated a significant increase in the affinity of pump sites for ouabain in aortic strips from hypertensive rabbits compared with those from normotensive rabbits (Scatchard analysis). The maximal binding capacity of the binding sites was similar in the two groups of animals, whereas the concentration of ouabain at which half-maximal binding occurred was lower in the aortic strips from hypertensive rabbits. Aortic strips from hypertensive rabbits showed greater sensitivity to the relaxant effect of potassium after incubation in potassium-free solution. The potassium-induced relaxation in aortic strips from hypertensive rabbits was more sensitive to the inhibitory effect of ouabain than that in strips from normotensive rabbits. These results suggest that the increased sensitivity to potassium and ouabain in vascular smooth muscle from hypertensive rabbits is due to an increased affinity of the pump sites for these substances. PMID- 6293320 TI - Elimination characteristics of disulfiram over time in five alcoholic volunteers: a preliminary study. AB - The authors studied the elimination of disulfiram and its metabolites for 24 hours after disulfiram administration in five healthy male alcoholic volunteers. Using high-performance liquid chromatography, they found that a single 500-mg dose resulted in a gradual increase in plasma disulfiram and its metabolites, with peak levels generally occurring 8 hours after dosing. There was considerable interpatient variability (e.g., in one volunteer no disulfiram was detected during the entire 24-hour sampling period). The authors also found that breath carbon disulfide was 9.1% of the dose of disulfiram administered, which is less than that expected theoretically. PMID- 6293321 TI - Psychological response of patients receiving two drug regimens for lung carcinoma. AB - Seventy-seven patients with small cell lung carcinoma were assigned randomly to two chemotherapy regimens to assess their psychological response to each regimen. One produced less depression and fatigue than the other, despite the absence of differences in tumor response. PMID- 6293322 TI - Data requirements to measure progress on the objectives for the nation in health promotion and disease prevention. AB - The Reagan Administration has adopted the policy guidelines developed over the previous few years in the disease prevention and health promotion initiative of the Carter Administration. Broad national consensus had been sought in the formulation of 226 measurable objectives for the decade. We classify the prevention objectives according to their position in an implied causal chain: 1) improved programs, 2) increased public and professional awareness, 3) reduced risk factors, and 4) improved health status. Prior to 1980, the data systems and periodic surveys sponsored by federal agencies and national organizations covered only four of the 42 objectives in the public and professional awareness category, whereas at least half of the objectives in each of the other three categories were covered by available national data sources, mostly federal. Sample surveys are needed to measure the majority of the currently unmeasured objectives in all four categories. Private and state health interview surveys are needed to supplement the federal capacity, especially in the face of federal cutbacks in survey capacity. PMID- 6293323 TI - Demonstration of alpha-1-antitrypsin and alpha-1-antichymotrypsin in fibrous histiocytomas using the immunoperoxidase technique. AB - Until recently, the diagnosis and classification of malignant fibrous histiocytomas (MFH) has been based on light- and electron-microscopic appearances. Tissue culture studies have led to the suggestion that these tumors have a common histiocytic origin. Using the immunoperoxidase PAP technique, a variety of soft-tissue tumors have been stained for the histiocyte markers alpha 1-antitrypsin (A1AT), alpha-1-antichymotrypsin (A1ACT) and lysozyme. A1AT and A1ACT are found to be useful specific markers for tumors of the MFH group whereas lysozyme is not a reliable marker for such tumors. The presence of these substances within the tumors supports the theory that they share a common origin from tissue histiocytes. Only a proportion of superficial skin histiocytomas stain for A1AT and A1ACT, raising the possibility that these are a heterogeneous group and do not share a common histogenesis with MFH. PMID- 6293324 TI - Ganglion cells in metastatic Wilms' tumor. Review of a histogenetic controversy. PMID- 6293325 TI - Antigenic relationships of flaviviruses with undetermined arthropod-borne status. AB - In contrast to most of the arthropod-borne flaviviruses, the flaviviruses with undetermined arthropod-borne status are probably disseminated only by direct contact with excreta (saliva, urine, feces, etc.); however, as yet undescribed arthropod transmission cycles may be found for some of them. Twenty-two of these flaviviruses, including prototype and recently isolated strains, were compared. Biologic properties were defined by infectivity titrations in suckling mice and Vero, LLC-MK2, and primary Pekin duck embryo cells, and antigenic relationships were defined by complement-fixation and plaque reduction neutralization tests. An antigenic classification scheme is proposed. Antigenic and biologic properties delimit two large clusters. The first, comprising a single antigenic complex, includes those which have yet to be isolated from arthropods, but are likely to be so (Israel turkey meningoencephalitis, Koutango, Negishi and Aroa viruses). The second, encompassing five antigenic complexes, is comprised of viruses which have been isolated exclusively from rodents or bats (Saboya, Carey Island, Dakar bat, Sokuluk, Bukalasa bat, Entebbe bat, Phnom Penh bat, Modoc, Sal Vieja, Jutiapa, San Perlita, Cowbone Ridge, Rio Bravo, Apoi, Tamana bat and Montana Myotis leucoencephalitis viruses) but includes three viruses (Saboya, Sokuluk and Entebbe bat viruses) which may be arthropod-borne, as indicated by replication in mosquito cells in vitro. PMID- 6293326 TI - Incidence of lobular carcinoma in bilateral breast cancer. AB - The histology and clinical records of 52 patients with bilateral breast cancer recorded in a community tumor registry were reviewed. Previous studies have demonstrated the propensity of lobular carcinoma to occur bilaterally. This view is supported by the large number of lobular cancers found in our patients. Thirty six percent of the patients with bilateral disease had lobular cancer in at least one breast. Those with lobular cancer tended to be younger and more likely to have simultaneous cancers than did patients with nonlobular carcinoma. In those patients in whom the occurrence of tumors was not simultaneous, they were smaller in the second breast but had similar rates of axillary metastases. This study raises the question of how best to manage the contralateral breast in patients with breast cancer. Lobular carcinoma is one marker of the likelihood for development of disease in the second breast; but if advantage is to be gained by this finding, investigation of the opposite breast is best done early. Finally, thorough examination of patients with nonlobular carcinoma must not be ignored because they still comprise the majority of bilateral breast cancers. PMID- 6293327 TI - Ion transport in the endolymphatic space. AB - The endolymph differs from all other extracellular fluids in its positive potential and in the ionic compositions in the various parts of the endolymphatic space. The endolymphatic space is surrounded by a tight neuroepithelium. The strial cells, which have a negative intracellular potential of -67 mV, generate a positive endolymphatic potential of +80 mV. The potassium ions are pumped into the endolymph by an active energy-consuming process. Intracellular ion concentrations of the marginal strial cells are 135 mM for potassium and 6 mM for sodium, which are comparable to concentrations in other epithelial cells. During anoxia or after ethacrynic acid injection potassium concentrations in the cochlear endolymph and in the marginal strial cells decrease, while sodium concentrations increase. PMID- 6293328 TI - Adenylate cyclase and cochlear fluid balance. PMID- 6293329 TI - Cellular localization of Na+,K+-ATPase in the mammalian cochlear duct: significance for cochlear fluid balance. AB - Cytochemical and autoradiographic procedures were employed to determine the cellular distribution of Na+,K+-ATPase in the guinea pig cochlea. The highest activity was associated with the stria vascularis and was restricted almost entirely to the contraluminal extensions of the marginal cells. Elevated levels of activity were also observed in stromal cells of the spiral prominence, external sulcus, and spiral limbus. The pattern of activity in the latter tissues was unusual in being symmetrically distributed along the plasma membranes of the reactive cells. In the organ of Corti, only neural elements showed appreciable activity. Possible functions of the enzyme are discussed in relation to cochlear fluid balance. PMID- 6293330 TI - Possible functional roles of Na+,K+-ATPase in the inner ear and their relevance to Meniere's disease. AB - This article reviews the functions of the enzyme Na+,K+-ATPase in epithelial tissues and discusses early and recent biochemical, physiologic and morphologic studies of the enzyme in the inner ear. The purpose of the investigation was to learn whether a relationship between perturbations in activity of the enzyme and Meniere's disease is possible. It is concluded that the preponderance of the evidence indicates that Na+,K+-ATPase plays a role in regulating ion transport into the scala media, but that the significance of the distribution of the enzyme along only one cell type (the marginal) in the functional chains of cells of the outer cochlear wall needs further study. The possible vasoconstrictive effects of ouabain perfusions employed by some investigators must also be taken into account. Recent cytochemical and autoradiographic studies have demonstrated high levels of Na+, K+-ATPase on cochlear nerve fibers, especially near the foramina nervosa and within the organ of Corti. Thus, perturbations in Na+,K+-ATPase activity in the inner ear not only could affect certain aspects of fluid balance, but also could account for the sensory disturbances experienced by patients who have Meniere's disease. PMID- 6293331 TI - Evidence for a perilymphatic origin of the endolymph: application to the pathophysiology of Meniere's disease. AB - The origin of the endolymph was elucidated by kinetic studies of the entry of water and electrolytes into endolymph and perilymph after intravenous administration of radioactive tracers in rats. The compartmental analysis of the data and the comparison of this study with the results of Konishi and associates (Acta Otolaryngol (Stockh) 86, 22-34 and 176-184, 1978), using perilymphatic perfusion of tracers, indicate that perilymph rather than plasma may be considered the precursor of endolymph. Since the cochlear epithelium was found to be freely permeable to water, an alteration of electrolyte transportation across the membranous labyrinth may be involved in the pathophysiology of Meniere's disease. Chloride transport across the cochlear epithelium was investigated using acetazolamide, a specific carbonic anhydrase inhibitor. PMID- 6293332 TI - Giant granules and rod-shaped inclusions arising de novo in monocytes and macrophages cultured from a patient with acute monocytic leukemia. AB - In this paper we describe the de novo appearance of azurophilic giant granules and rod-shaped inclusions in monocytes and macrophages grown from the peripheral blood and bone marrow of a patient with acute monocytic leukemia; no such inclusions were evident in direct smears of the patient's peripheral blood or bone marrow. The cytochemical profile of the inclusions, their origin in mononuclear phagocytes only, and their development exclusively in vitro suggest that they are distinct from Auer rods, Chediak-Higashi-like giant granules, and other abnormal azurophilic inclusions previously described in patients with acute non-lymphocytic leukemia. PMID- 6293333 TI - Desmoplastic malignant melanoma: a study by conventional and electron microscopy. AB - A 61-year-old white man presented himself with a mass that had recurred on the chin. Conventional microscopy of sections from this mass showed atypical spindle cells surrounded by abundant collagen and resembled fibrosarcoma. Stains for melanin revealed pigment in the cytoplasms of the atypical spindle cells. Study of sections by electron microscopy demonstrated round, oval, and spindle-shaped cells having desmosomes and containing abundant melanosomes in varying stages of maturation. On the basis of both conventional microscopy and electron microscopy, the diagnosis was desmoplastic malignant melanoma. An unusual finding by electron microscopy was the presence of tubuloreticular intracytoplasmic inclusions in the neoplastic melanocytes. PMID- 6293334 TI - [Onset of effect of an i.v. ranitidine bolus]. AB - Ranitidine 50 mg or 0.9% saline was given i.v. to 12(6) volunteers and pH and volume of the gastric aspirate was measured in short-time intervals over a period of 6 hours. There was an increase in pH above 4 in all subjects starting within 30 min after ranitidine injection. Concomitantly volume secretion was reduced to 35-50%. i.v. administration of ranitidine 30-45 min before induction of anaesthesia should, therefore, reduce the number of patients theoretically at risk of acid-induced pulmonary damage. PMID- 6293335 TI - A redox equilibrator for the preparation of cytochrome oxidase of mixed valence states and intermediate compounds for X-ray synchrotron studies. PMID- 6293336 TI - Lowry protein determination on membrane preparations: need for standardization by amino acid analysis. PMID- 6293337 TI - Sensitive and inexpensive dual-wavelength reflection spectrophotometry using interference filters. PMID- 6293338 TI - Automated proton titrations of pancreatic phospholipase A2. PMID- 6293339 TI - Resolution of optical isomers by chiral high-performance liquid chromatography: separation of dihydrodiols and tetrahydrodiols of benzo[a]pyrene and benz[a]anthracene. PMID- 6293340 TI - Affinity chromatography of hyaluronate on glutaraldehyde-fixed SV-3T3 cells. PMID- 6293341 TI - SV40 RNA: filter hybridization for rapid isolation and characterization of rare RNAs. PMID- 6293342 TI - Comparison of extraction methods for determination of polybrominated biphenyl residues in animal tissue. PMID- 6293343 TI - Concanavalin A receptor sites on lymph node cells in vivo and in vitro. AB - The distribution and density of receptors for concanavalin A (Con A) on the surfaces of cells of intact and isolated popliteal and axillary lymph nodes were investigated in the rabbit. Intact lymph nodes were perfused via the subcapsular (marginal) sinus with either Con A peroxidase or Con A ferritin, fixed with glutaraldehyde, and processed for electron microscopy. Both Con A peroxidase and Con A ferritin were distributed on the plasmalemma of lymphocytes, macrophages, neutrophils, plasma cells, reticular endothelial cells, and the vascular endothelium. Counts of Con A-conjugated ferritin particles indicated that the density of Con A receptors was generally similar for lymphocytes, macrophages, and neutrophils but lower on plasma cells. When lymph node cells were isolated by mechanical methods and exposed to Con A ferritin, the label was homogenously distributed on the cell surfaces of most cells. However, Con A binding was significantly higher on the surface of isolated cells than in the intact node. It is suggested that the increase in density of Con A binding sites on isolated cells may possibly be due to an unmasking of cell surface moieties in which additional Con A receptor sites become available as a result of the isolation procedure. The density of Con A ferritin binding sites was also significantly lower on the surface of isolated plasma cells than the lymphocyte and macrophage, suggesting that the density distribution of cell surface saccharides is different for various lymphoid cells. PMID- 6293344 TI - Effects of fentanyl on the response of plasma beta-endorphin immunoreactivity to surgery. AB - Beta-endorphin appears to play a definite role in the biologic response to stress and in the endogenous mechanism of pain perception. Opiates exogenously administered during surgery decrease or even suppress the activation of "stress hormones" such as ACTH and cortisol. In the present study, the authors tried to assess the effects of fentanyl administration on plasma beta-endorphin immunoreactivity PBE(ir) during surgical stress. In one group of nine patients, a standard enflurane-based general anesthetic technique without opiates was used for a staging laparotomy. A second group of ten patients undergoing the same type of surgery received fentanyl (10-20 micrograms/kg) as the primary anesthetic drug. In both groups, multiple blood samples were collected prior to, during, and after surgery, following the same time protocol. PBE(ir), plasma cortisol and, in five patients, plasma ACTH were determined by radioimmunoassay. There was no significant change in PBE(ir) in either group after anesthetic induction. Unlike the enflurane group, the fentanyl group did not demonstrate any significant increase from baseline in PBE(ir) during surgery. There was a significant group difference between enflurane and fentanyl in PBE(ir) levels for both "early" and "late" surgery values, but not for the "awake" values (recovery period) where both groups had elevated PBE(ir) levels. Plasma cortisol and plasma ACTH changes followed a trend similar to those of PBE(ir). The suppression of both cortisol and PBE(ir) responses during surgery after administration of fentanyl provides further evidence for the involvement of the endorphin system in the stress response and for its physiologic association with the hypothalamo pituitary axis. PMID- 6293346 TI - More information about pipecurium, a new neuromuscular blocking agent. PMID- 6293345 TI - The effects of halothane on sympathetic ganglionic transmission. AB - The effects of halothane on ganglionic transmission were studied in the stellate ganglion of the guinea pig using intracellular recordings in vitro. Depression of synaptic transmission is one of the actions common to many general anesthetics. The aim of this study was to investigate which of the processes involved in synaptic transmission are affected by halothane in concentrations comparable to those used during surgical anesthesia. The neurons of the stellate ganglion were depolarized using preganglionic nerve stimulation, postganglionic nerve stimulation, and intracellular stimulation before ad after introduction of halothane (vaporizer settings of 0.75% and 1.5% produced bath concentrations of 8 and 18 mg/dl, respectively). Halothane at both concentrations depressed sympathetic ganglionic transmission which was induced by stimulation of preganglionic nerves. Axonal transmission and the excitability of the postganglionic neurons to direct intracellular stimulation was far less sensitive to halothane than synaptic transmission. The depression of ganglionic transmission seen in the present study was most likely due to a decrease in transmitter release although alterations in postsynaptic receptor properties could have been involved as well. The decrease in sympathetic activity resulting from depression of ganglionic transmission probably contributes to the arterial hypotension seen during halothane anesthesia, along with direct myocardial depression, inhibition of catecholamine release from the adrenal medulla, direct action on vascular smooth muscle, and central sympathetic depression. PMID- 6293347 TI - Long-term succinylcholine infusion during isoflurane anesthesia. AB - The characteristics of the neuromuscular blockade produced by prolonged succinylcholine infusion were compared in 40 patients anesthetized with either nitrous-oxide-isoflurane (0.75-1.50% inspired) or nitrous-oxide-fentanyl. Neuromuscular transmission was monitored using train-of-four stimulation and the infusion rate was adjusted to keep the first twitch at 10-15% of its control value. Initially, all patients exhibited a depolarizing-type block, and the infusion rates were similar in the isoflurane (61 micrograms . kg-1 . min-1) and fentanyl (57 micrograms . kg-1 . min-1) groups. Tachyphylaxis developed in both groups and correlated well with the onset of non-depolarizing (phase II) block. Both occurred sooner and at a lower cumulative dose in the isoflurane groups. After 90 min, infusion rates were similar in both groups (isoflurane: 107 micrograms . kg-1 . min-1, fentanyl;: 93 micrograms. kg-1 . min-1). After the infusion was stopped, the recovery of the train-of-four ratio was inversely related to the dose and duration of exposure to succinylcholine, and was slower with nitrous-oxide-isoflurane anesthesia. After 10 min of recovery, patients receiving isoflurane exhibited train-of-four ratios of 0.5 or less after 8.5 mg/kg succinylcholine and 103 min. Corresponding figures for fentanyl patients were 13 mg/kg and 171 min. The block in all 13 patients (eight with isoflurane, five with fentanyl) who did not recover spontaneously was antagonized successfully with atropine and neostigmine. It was concluded that with succinylcholine infusion of 90 min or less, isoflurane accelerates the onset of tachyphylaxis and phase II neuromuscular block without affecting succinylcholine requirements. These results, with isoflurane, were similar to those reported previously with enflurane or halothane. PMID- 6293348 TI - Duration of colostral antibodies to bovine leukemia virus by two serologic tests. AB - The duration of detectable colostral antibodies to the glycoprotein antigen of bovine leukemia virus was studied in calves which were born to bovine leukemia virus-infected cows, but showed no serologic evidence of prenatal infection. Colostral antibodies detectable by an agar-gel immunodiffusion test (AGIT) persisted for less than 1 month to 6 months (mean 2.9 months) in the 139 calves examined. Colostral antibodies were detectable 1 to 5 months longer by radioimmunoprecipitation assay than by the AGIT in 22 of the 24 calves studied comparatively. The mean duration of colostral antibodies in those 24 calves was 3.8 months (min-max, 2 to 6 months) for the AGIT and 6.0 months (min-max, 4 to 9 months) for the radioimmunoprecipitation assay. PMID- 6293349 TI - Virulence and in vitro growth of a cell-adapted strain of equine infectious anemia virus after serial passage in ponies. AB - Five serial passages of a cell-adapted strain of equine infectious anemia (EIA) virus were conducted in Shetland ponies. The 13 recipient ponies became agar-gel immunodiffusion test-positive by 25 days after they were inoculated. The virulence of the cell-adapted strain of EIA virus markedly increased through 3 serial passages, although individual variation within passages was high. The 1st serial-passage recipient remained afebrile through 200 days, whereas a febrile episode occurred about every 185, 44, 35, and 33 days in the 2nd, 3rd, 4th, and 5th serial-passage recipients, respectively. Severe clinical signs of EIA were noted in the ponies at each serial passage, but the mean virulence rating of each passage, graded on frequency of febrile episodes and number of clinical signs evident within 200 days after ponies were inoculated, increased from 0 through 4, 21, 24, and 29 for the 1st through 5th serial passages, respectively. Isolates of EIA virus, made in fetal equine kidney cells, were obtained from plasma of 75% of the samples of blood collected during febrile episodes and from 45% of the samples collected during afebrile periods, indicating that the cell culture growth capacity of this strain of EIA virus may be relatively stable through 5 serial passages in Shetland ponies. PMID- 6293350 TI - Effect of infectious bovine rhinotracheitis virus immunization on viral shedding in challenge-exposed calves treated with dexamethasone. AB - Calves not vaccinated with infectious bovine rhinotracheitis virus (IBRV) became latently infected when challenge exposed and treated with dexamethasone (DM). Calves that shed IBRV after DM treatment were considered to be latently infected. Vaccination with a temperature-sensitive intranasal vaccine or with formalinized IBRV in Freund's complete adjuvant (IBRV-FCA) protected some, but not all, calves against latent infection--indicating a role for the immune response in preventing latent infection. That all latently infected calves were not detected after DM treatment was indicated by the fact that after a 2nd DM treatment of 3 calves treated 6 months previously and not found to shed virus, 1 of the calves was latently infected. Latently infected calves were inoculated with successive doses of IBRV-FCA and treated with DM. Nonvaccinated calves shed virus, whereas vaccinated calves similarly treated did not shed virus. Because both groups had a comparable cell-mediated immune response, as determined by blastogenic response to IBRV, but the vaccinated group had significantly higher virus-neutralizing antibody titers, a role for humoral antibody in preventing viral shedding was indicated. PMID- 6293351 TI - The role of transbronchial biopsy for the diagnosis of diffuse pneumonias in immunocompromised marrow transplant recipients. AB - We studied the use of transbronchial biopsy for the diagnosis of diffuse pneumonia in marrow transplant recipients. Transbronchial biopsy results were directly compared with open-lung biopsy results by performing the procedures simultaneously in the same lobe of the lung and processing the specimens in parallel. There were 24 cases of pneumonia diagnosed in 22 patients. Transbronchial biopsy correctly identified 3 of 5 cases of Pneumocystis carinii and none of the 5 cases of viral pneumonia. The overall sensitivity of transbronchial biopsy was 58%, with a 13% incidence of moderate hemorrhage and no deaths. We conclude that the open-lung biopsy remains the procedure of choice for the diagnosis of acute, diffuse pneumonia in the immunocompromised marrow transplant recipient. PMID- 6293352 TI - Survival in lung cancer. An analysis of the effects of age, sex, resectability, and histopathologic type. AB - Records of 6,686 patients with primary lung malignancies were retrieved from a tumor registry. Survival curves according to histopathologic type demonstrated that patients with anaplastic tumors have a lower cumulative survival than those with either squamous cell cancers or adenocarcinomas (p less than 0.001), but survival for the latter two histologic types is similar. Age at diagnosis influences survival, and women with squamous or anaplastic tumors have a better prognosis than men with the same cell types (p less than 0.001). Although selection for resection is strongly associated with lower early mortality, all patients alive 2 yr after diagnosis have a similar prognosis is regardless of initial resectability. Each survival curve shows a high early mortality and a lower late mortality, indicating that each group can be considered to consist of 2 subgroups with different risks of dying of lung cancer. A mathematical model is described that closely approximates the actuarial survival. PMID- 6293353 TI - Local cellular immunity in tuberculous pleurisy. AB - The characteristics and function of lymphocytes in both pleural exudate and peripheral blood in 18 patients with tuberculous pleurisy were studied. The pleural fluid of 13 of 16 patients with tuberculous pleurisy had more T lymphocytes than the peripheral blood. The difference between them was significant by the t test (p less than 0.05). When lymphocytes in peripheral blood were cocultured with purified protein derivative (PPD), lymphocytes from only 4 of 18 patients produced immune interferon on the fifth day. The titer was only 4 or 8 units/ml. On the contrary, lymphocytes in pleural fluid from 17 of 18 patients reacted to PPD and produced immune interferon on the fifth day, and titers were much higher (more than 128 units/ml) than those of lymphocytes in peripheral blood. When the relationship between the interferon titer produced by lymphocytes in pleural effusion and tuberculin skin reaction was investigated, the group with the stronger skin reactions showed a significantly higher interferon production (p less than 0.05). In conclusion, exudative-sensitized lymphocytes in morbid sites reacted to the specific antigen more effectively and produced titers of lymphokines than circulating lymphocytes. PMID- 6293354 TI - Experimental diffuse alveolar damage in baboons. PMID- 6293355 TI - [Congenital gangrene of the right upper limb]. PMID- 6293356 TI - Herpes simplex virus pneumonia: clinical, virologic, and pathologic features in 20 patients. AB - Herpes simplex virus (herpesvirus) was isolated from autopsy lung specimens of 20 patients with clinical, roentgenographic, and histologic evidence of pneumonia. Mucocutaneous herpesvirus infection preceded the onset of pneumonia in 17. Twelve patients had focal pneumonia, 10 of whom had concomitant herpetic tracheitis, esophagitis, or both. Eight patients had diffuse interstitial pneumonia, six of whom had dissemination of herpesvirus to the other organs. Of the eight lung isolates available for typing, seven were herpesvirus-1 and one, herpesvirus-2. A high prevalence of herpesvirus antibody in serum samples obtained before pneumonia and identical restriction endonuclease patterns between mucosal and lung isolates in individual patients indicated that, in most cases, herpesvirus pneumonia was due to endogenous reactivation of virus. Focal herpesvirus pneumonia appeared to result from contiguous spread of herpesvirus to lung parenchyma, whereas diffuse interstitial pneumonia appeared to be a manifestation of hematogenous dissemination of virus. PMID- 6293357 TI - Inflammatory fibrous histiocytoma: an important variant of malignant fibrous histiocytoma highly responsive to chemotherapy. AB - Inflammatory fibrous histiocytoma is a recently recognized variant of malignant fibrous histiocytoma. Patients managed with surgical excision or radiation therapy usually have had multiple recurrences, often with metastases. The disease is insidious but ultimately fatal. Four consecutive patients were treated with inflammatory fibrous histiocytoma with alkylating agents with or without anthracyclines and produced prolonged and sustained remissions. Inflammatory fibrous histiocytoma may be another highly chemotherapeutically responsive tumor that deserves active case identification for aggressive curative therapy. PMID- 6293358 TI - Extragonadal germ-cell tumors. PMID- 6293359 TI - Glucagonoma and dacarbazine. PMID- 6293360 TI - Ketoconazole in systemic mycoses. PMID- 6293362 TI - An update on antioxidant theory: spin trapping of trichloromethyl radicals in vivo. PMID- 6293361 TI - Treatment of systemic mycoses with ketoconazole: emphasis on toxicity and clinical response in 52 patients. National Institute of Allergy and Infectious Diseases collaborative antifungal study. AB - The pharmacology, in vitro mycologic activity, toxicity, and efficacy of ketoconazole were studied in a Phase-II evaluation by the National Institutes of Health and National Institute of Allergy and Infectious Disease Mycoses Study Group. This report emphasizes the toxicity and clinical response data in 52 patients with the following systemic mycoses: blastomycosis in 16 patients; nonmeningeal coccidioidomycosis in 13; histoplasmosis in 8; nonmeningeal cryptococcosis in 7; sporotrichosis in 7; and both blastomycosis and nonmeningeal coccidioidomycosis in 1. Maximum daily doses of ketoconazole were 100 mg in 1 patient; 200 mg in 23; 400 mg in 12; and 600 mg in 16. In 52% of the patients, duration of therapy ranged from less than 1 to 6 months, whereas in 35%, duration ranged from 7 to 12 months, and in 13%, from 12 to 22 months. In 35 patients (67%), evidence of toxicity was not seen. Nausea, anorexia, or vomiting occurred in 21%. Cure or marked improvement was shown in 27 patients (52%), whereas failure of the primary course was seen in 14 (27%) and relapse after ketoconazole was discontinued in 11 (21%). Although this evaluation did not provide clear-cut clinical response data, our results indicate that ketoconazole, in the dosage regimens used, was more effective in patients with histoplasmosis and nonmeningeal cryptococcosis than in patients with blastomycosis and nonmeningeal coccidioidomycosis, and least effective in patients with sporotrichosis. PMID- 6293364 TI - Alterations of enzymes in the red blood cell membrane in vitamin E deficiency. PMID- 6293363 TI - The influence of vitamin E on human polymorphonuclear cell metabolism and function. AB - These studies on the effect of administration of 1,600 units of vitamin E to humans indicated the following responses to the PMNs (TABLE 6). Functional alterations occur with an increased ability to ingest particles but a mild decrease in bactericidal potency of the PMN. Although the respiratory burst is slightly enhanced as is superoxide anion release, H2O2 release from the PMN is markedly impaired. The hexose monophosphate shunt activity, which is dependent on intracellular H2O2 is decreased during phagocytosis. Membrane responses such as changes in order parameter during phagocytosis as reported by the stearic acid analogue probe 5DS are similar to those of normal PMNs. The release of arachidonic acid from membranes of vitamin E PMNs during phagocytosis of opsonized zymosan is slightly enhanced, indicating normal phospholipase A2 activation. NADH oxidase-derived H2O2 is not impaired within phagocytic generated by NADPH oxidase in phagocytic vesicles, accounting for impairment in HMPS activity and bactericidal activity in these cells. PMID- 6293365 TI - Mechanism of action of vitamin E on platelet function. PMID- 6293366 TI - Demonstration of specific binding sites for 3H-RRR-alpha-tocopherol on human erythrocytes. AB - Previous work from our laboratory demonstrated specific binding sites for 3H-RRR alpha-tocopherol (3H-d alpha T) in membranes of rat adrenal cells. As tocopherol deficiency is associated with increased susceptibility of red blood cells to hemolysis, we investigated tocopherol binding sites in human RBCs. Erythrocytes were found to have specific binding sites for 3H-d alpha T that exhibited saturability and time and cell-concentration dependence as well as reversibility of binding. Kinetic studies of binding demonstrated two binding sites--one with high affinity (Ka of 2.6 x 10(7) M-1), low capacity (7,600 sites per cell) and the other with low affinity (1.2 x 10(6) M-1), high capacity (150,000 sites per cell). In order to localize the binding sites further, RBCs were fractionated and greater than 90% of the tocopherol binding was located in the membranes. Similar to the findings in intact RBCs, the membranes exhibited two binding sites with a respective Ka of 3.3 x 10(7) M-1 and 1.5 x 10(6) M-1. Specificity data for binding demonstrated 10% binding for RRR-gamma-tocopherol, but not other tocopherol analog exhibited competition for 3H-d alpha T binding sites. Instability data suggested a protein nature for these binding sites. Preliminary studies on Triton X-100 solubilized fractions resolved the binding sites to a major component with an Mr of 65,000 and a minor component with an Mr of 125,000. We conclude that human erythrocyte membranes contain specific binding sites for RRR-alpha-tocopherol. These sites may be of physiologic significance in the function of tocopherol on the red blood cell membrane. PMID- 6293367 TI - Chemodectomas (nonchromaffin paragangliomas) of the glomus jugulare, glomus vagale and carotid body. PMID- 6293368 TI - Infection in neutropenia: Anti-endotoxin effect of polymyxin. PMID- 6293369 TI - Fibronectin in human astrocytomas grown in tissue culture. PMID- 6293370 TI - Ross River virus-induced demyelination: I. Pathogenesis and histopathology. AB - Ross River virus (strain T48) infection in mice causes an encephalomyelitis characterized by focal, primary demyelination in the cerebellum, brain stem, and spinal cord. Maximal serum and brain content of virus occurs on days 2 and 4, respectively. Virus is not detectable in serum after day 3 or in brain after day 9. Histopathological lesions are present by day 2 and consist of perivascular macrophage and polymorphonuclear leukocyte infiltration, focal necrosis in the internal granule cell layer, and myelin disruption. Mononuclear cell infiltrates are present by day 5. Foci of demyelination in the presence of preserved axons become more widespread by day 8, and early partial remyelination occurs by day 13. Immunosuppression reduces the mononuclear cell infiltration but does not alter the demyelination . Although the mechanism of Ross River virus-induced demyelination is not known, these findings suggest that it is not immune mediated. PMID- 6293371 TI - Microbial envelope proteins related to iron. PMID- 6293372 TI - Low-molecular-weight enzyme inhibitors of microbial origin. PMID- 6293373 TI - [Effect of various carbon and nitrogen sources on the biosynthesis of ristomycin, protease and pigments by a culture of Nocardia fructiferi var. ristomycini]. AB - The effect of various sources of carbon and nitrogen on the biosynthesis of ristomycin, protease and pigments by Nocardia fructiferi was studied. It was shown that the carbon sources had the most significant effect on the biosynthesis of the antibiotic. The maximum biosynthetic activity of the Nocardia was observed in the medium containing 1-2 per cent of soybean meal and 2 per cent of glycerol. Under such conditions all the three biologically active substances formed. The contents of ristomycin, protease and pigments amounted to 562-649 microgram/ml, 26-30 PU/ml and 0.45-0.63 conditional units, respectively. PMID- 6293374 TI - [Use of luminescent analysis for studying the characteristics of the virus inhibiting action of polyene antibiotics]. AB - The inhibitory effect of flavopentin was shown on the models of infectious and oncogenic viruses. Flavopentin is a polyenic antibiotic belonging to pentaenes. The characteristic features of the polyene interaction with the plasma membrane of the host cell were defined with the use of the luminescence analysis, when the antibiotic was applied with the therapeutic or prophylactic purposes in experimental influenzal infection. PMID- 6293375 TI - Evaluation of the anti-herpesvirus drug combinations: virazole plus arabinofuranosylhypoxanthine and virazole plus arabinofuranosyladenine. AB - Combinations of Virazole plus arabinofuranosylhypoxanthine (ara-Hx) and Virazole plus arabinofuranosyladenine (ara-A) were investigated in KB or BHK cells infected with types 1 or 2 herpes viruses. Combinations of Virazole and ara-Hx exhibited significant synergy as evaluated graphically (isobolograms) or by fractional inhibitory concentration (FIC) indices. Optimal ratios for the combination were 1:1 to 1:10 for Virazole to ara-Hx. At these ratios, FIC indices in the range of 0.5-0.2 were commonly observed. Combinations of Virazole and ara A were antagonistic when observed in the presence of pentostatin, an adenosine deaminase inhibitor. In the absence of pentostatin, the minimum inhibitory concentration (MIC) of ara-A and degree of synergy with Virazole were variable. PMID- 6293376 TI - Comparison of the in vitro and in vivo anti-herpes activities of 1-beta-D arabinofuranosylthymine and its 5'-monophosphate. AB - In vitro and in vivo anti-herpes activities of 1-beta-D-arabinofuranosylthymine 5'-monophosphate (ara-TMP) were compared with those of 1-beta-D arabinofuranosylthymine (ara-T). On a molar basis ara-TMP was almost as active as ara-T against six strains of herpes simplex virus type 1 (HSV-1) and type 2 (HSV 2) as monitored by a cytopathogenicity-inhibition and a plaque reduction assay in human embryonic lung fibroblast cells. When tested against experimental encephalitis in mice inoculated intracerebrally with HSV-1, intraperitoneal or intravenous treatment with 150 mg/kg/day of ara-TMP or 100 mg/kg/day of ara-T, for 5 days was effective in increasing in the mean survival time of mice. For a single dose of ara-TMP, intravenous administration was more effective than intraperitoneal or oral administration. However, oral administration of ara-T was the most effective of the treatment regimens used. Substantial plasma levels of ara-T were detected for a longer time after oral administration of ara-T than after intravenous administration of ara-TMP or ara-T, suggesting that the efficacy of oral administration of ara-T may be correlated with the maintenance of the substantial blood drug levels. PMID- 6293377 TI - Synergistic activities of type I (alpha, beta) and type II (gamma) murine interferons. AB - Type I (alpha, beta) and type II (gamma) murine interferons are able to potentiate each other with respect to the inhibition of encephalomyocarditis (EMC) virus and of herpes simplex virus type 1 (HSV-1) multiplication in a murine cell line (DBT). Examination of two double-stranded RNA-dependent enzymes in DBT cells, the 2-5A synthetase and the 67,000 MW protein phosphokinase indicates that mixed interferon preparations act synergistically at least with respect to an increase in the activity of the former enzyme. The results obtained with gamma interferons of different origin and of different specific activity suggest that interferon itself, rather than the lymphokines present in the interferon preparations, is responsible for the synergistic effect. PMID- 6293379 TI - Phosphorylation of glycogen synthase by cyclic AMP-independent glycogen synthases kinase-1 (GSK-1) comparative study with cyclic AMP-dependent protein kinase and phosphorylase kinase. PMID- 6293381 TI - Catalytic and kinetic properties of purified high-affinity cyclic AMP phosphodiesterase from dog kidney. PMID- 6293380 TI - Characterization of calmodulin-mediated phosphorylation of cardiac muscle sarcoplasmic reticulum. PMID- 6293383 TI - Reaction of iron-EDTA chelates with the superoxide radical. PMID- 6293382 TI - Equilibrium constants under physiological conditions for the reactions of the nonphosphorylated pathway of L-serine biosynthesis. PMID- 6293385 TI - A deuteron and proton magnetic resonance relaxation study of beta-lactoglobulin a association: some approaches to the Scatchard hydration of globular proteins. PMID- 6293384 TI - Sodium-dependent activation of intestinal brush-border sucrase: correlation with activation by deprotonation from pH 5 to 7. PMID- 6293386 TI - Multiple expressions of the activity of guanine nucleotide regulatory protein in human thyroid adenylate cyclase. PMID- 6293387 TI - Small cell carcinoma of the lung with Leser-Trelat sign. PMID- 6293388 TI - Endocrine function, morbidity, and mortality after surgery for craniopharyngioma. AB - The records of 59 children with craniopharyngioma first treated between 1960 and 1980 were reviewed. There was a high incidence of postoperative growth hormone deficiency (43 of 43), andrenocorticortopin deficiency (26 of 36), thyrotrophin deficiency (13 of 20), gonadotrophin deficiency (15 of 16), and diabetes insipidus (44 of 58). Fifteen patients have died: diabetes insipidus was a contributory factor in 5 patients who died within the first 4 months of surgery; and 8 further children died unexpectedly after the postoperative period, probably as a result of anterior pituitary insufficiency. In addition, hypopituitarism led to medical emergencies in 12, 9 of whom had symptomatic hypoglycaemia. These findings illustrate the high incidence of pituitary deficits which follow current methods for treating childhood craniopharyngioma, and the long-term risks associated with these deficits. PMID- 6293389 TI - Importance of catecholestrogens in the regulation of the ovarian cycle. PMID- 6293390 TI - [A case of cystic nephroblastoma]. PMID- 6293391 TI - The diagnostic value of testing for occult blood. AB - An analysis of 270 patients undergoing microdochectomy for nipple discharge from a single identifiable duct, and without an associated lump, revealed that occult cancers were all accompanied by hemoglobin positive discharge. A simple method of detecting the presence of hemoglobin in the discharge is described. The incidence of cancer was found to be 5.9%. In order of frequency, intraduct papilloma, duct ectasia, cystic disease, and carcinoma accounted for over 90% of the cases. Six patients in whom no malignancy was seen in the microdochectomy specimen subsequently developed cancer in the ipsilateral breast. A close follow-up of all patients with hemoglobin positive discharge and in whom no cause for the discharge is demonstrated at microdochectomy is stressed. PMID- 6293392 TI - Histoplasma infection of abdominal aortic aneurysms. AB - Fungal endarteritis resulting from progressive disseminated histoplasmosis may cause arterial aneurysms, or lead to infection of pre-existing aneurysms. Three patients with Histoplasma capsulatum infections of abdominal aortic aneurysms are reported. All had previous disseminated histoplasmosis and atherosclerotic peripheral vascular disease. All were considered cured of systemic infection when their aneurysms were discovered. Atherosclerotic vascular lesions may become infected during the course of systemic fungal disease and may serve as a haven for viable organisms in patients whose dissemination recurs despite seemingly adequate antifungal therapy. In treating these patients, resection of all infected arterial tissue, revascularization through uninfected tissues, and long term antimicrobial therapy are recommended. PMID- 6293393 TI - Bronchial carcinoma and the lymphatic sump: the importance of bronchoscopic findings. AB - The lymphatic sump of Borrie is an important area of regional node metastasis in each lung. The sump area is of increased importance on the right side, since bilobectomy has been recommended to ensure complete removal of lymphatic disease in patients with lower or middle lobe carcinoma. The role of bronchoscopy in assessing lymphatic metastasis of bronchial carcinoma was investigated in 42 patients at the North Carolina Memorial Hospital. Because of the high incidence of associated lymphatic metastases, bilobectomy is indicated for right lower or middle lobe lesions observed at bronchoscopy. Simple lobectomy may be utilized when bronchoscopic findings are negative and when there is no involvement of the lymph nodes of the sump of Borrie at the time of operation. If there is gross nodal involvement of the lymphatic sump, pneumonectomy will be required. PMID- 6293394 TI - Acute carcinoid syndrome: a complication of flexible fiberoptic bronchoscopy. AB - We report the case of a patient with functioning bronchial carcinoid tumor found as a solitary nodule. Acute carcinoid syndrome developed during transbronchial biopsy, and life-threatening hypotension occurred during operation. We emphasize that carcinoid tumors should be considered in the differential diagnosis of solitary nodules, and such a complication should be kept in mind when endobronchial or transbronchial biopsy is performed. PMID- 6293395 TI - Prolactin in seminal fluid. AB - Prolactin appears to modulate male reproduction by regulating hormone receptor levels in the testis and accessory sex organs and by facilitating spermatozoal biochemical processes associated with capacitation. A large portion of immunoreactive prolactin (iPRL) seems to be transported from blood to semen via the accessory sex organs and takes several days to make this transition. The iPRL in semen is heterogeneous in size and a unique portion may be associated with spermatozoa. Data from several clinical studies of seminal fluid prolactin are inconsistent. The variability of patient populations classified as fertile or infertile, the differences in prolactin assays used, and the lack of information about sample processing have been major causes of this difficulty. PMID- 6293396 TI - Hepatitis A vaccine could follow 'B' immunization. PMID- 6293397 TI - Murein and lipopolysaccharide biosynthesis in synchronized cells of Escherichia coli K 12 and the effect of penicillin G, mecillinam and nalidixic acid. AB - The incorporation of radioactive N-acetyl-glucosamine into murein and lipopolysaccharide of synchronized cells of Escherichia coli K 12 was followed over 100 min in the presence of antibiotics. At 20 min intervals cell walls were prepared. Lipopolysaccharide and murein sacculi were isolated and the radioactivity was quantified in both polymers. Labelled, newly synthesized murein was characterized according to murein subunits linked to lipoprotein, and the degree of crosslinkage. Furthermore, murein subunits containing anhydromuramic acid were determined, permitting the calculation of the average glycan chain length. The results indicated that penicillin G at 30 micrograms/ml stimulated the incorporation of new murein subunits into sacculi followed by a sudden increase in lipopolysaccharide incorporation into the outer membrane. The degree of crosslinkage in murein synthesized in the presence of 30 micrograms/ml penicillin G was higher than in the control, and almost twice as high as in murein synthesized in the presence of 20 micrograms/ml nalidixic acid. Both antibiotics inhibited cell division at the concentrations indicated. Murein synthesized in the presence of 2 micrograms/ml mecillinam also showed higher crosslinkage. However, about twice as much anhydromuramic acid-containing subunits were observed as in the control. At the same time lipopolysaccharide incorporation into the outer membrane was stimulated two- to three-fold. PMID- 6293398 TI - [Methods of non-surgical treatment of pulmonary carcinoma: current status and report of cases in the advanced stage]. PMID- 6293399 TI - [Partially differentiated cystic nephroblastoma]. PMID- 6293400 TI - [Electron microscopic findings in the livers of long-term survivors among child tumor patients after polychemotherapy]. AB - In 14 children which were treated by combined antineoplastic chemotherapy a postexamination was performed after a mean survival time of 5 years and a post treatment-interval of two and a half years. In this study also liver biopsies were examined by electron microscopy. Ultrastructural changes of the hepatocytes must be explained as a progressive metabolic efficiency of these cells. There was no direction for a chronic damage of the liver parenchyma induced by the therapy. PMID- 6293401 TI - [Epidemiological basis for the organization of after care in female breast cancer in the capital of East Germany-Berlin in the period from 1968 to 1980]. AB - The basis for a scientific aftercare are such characteristics as recurrence and metastasis rates, lethality classified according to age-groups and duration of observation. Comparable data like 5-years-lethality by age-groups in 1968 and 1973 (female patients with cancer of the breast diagnosed in the capital of the G.D.R., Berlin), causes of death, cumulative relative survival rates, excess mortality are of importance for the follow-up intensity directed to this group of patients by the cancer centre, the general-practitioner and the out-patient oncological department. The most important task will still be early diagnosis of malignant tumours, because detection of curable stages will have better results and such patients need less intensive aftercare. PMID- 6293402 TI - Some factors of immunopathogenesis of infectious mononucleosis in children. AB - Sixty-eight children with the symptoms of infectious mononucleosis were examined. In 25 of them anti-EBV antibodies were found and in 10 heterophilic antibodies. In serum of 16 children sero-positive to EBV A, M, G, D immunoglobulins were determined as well as T and B lymphocytes. A statistically significant increase in the number of B-IgA, B-IgG and B-IgM lymphocytes, augmentation of T lymphocyte number and polyclonal hypergammaglobulinemia were detected. PMID- 6293403 TI - Lipofuscin in rabbit skin: Its occurrence after ocular herpes simplex infection. AB - Systemic infection, induced by intraocular inoculation of type 1 herpes simplex virus (HSV 1) in young rabbits, is accompanied by the appearance of autofluorescent pigments precisely in the midportion of their fur follicles. Histochemical and solubility reactions of the pigment led to its characterization as a lipofuscinlike pigment. Follicle fluorescence correlated with the severity of clinical symptoms; it was present in 100% of the follicles of rabbits that died but was found in only 30% to 50% of follicles of surviving animals. Similar fluorescence was also present in 10% to 20% of the follicles of uninfected rabbits. This autofluorescent material may be formed by the peroxidation of lipids from sebaceous gland secretions, but neither HSV 1 nor antigens could be found in the skin as possible initiators of this reaction. PMID- 6293404 TI - Mucinous adenocarcinoma and clear cell adenocarcinoma arising in separate ovaries. PMID- 6293405 TI - Metabolic and endocrine changes in spinal cord injury: III. Less quanta of sensory input plus bedrest and illness. AB - This is the fourth article (Part III) of a 4-part series which provides a comprehensive review and analysis of the pertinent literature published over the last 25 years on the metabolic and endocrine consequences of spinal cord injury (SCI). The studies on SCI patients reviewed in this article include reports and investigations concerning plasma ACTH, plasma and urine corticosteroids and their diurnal variations at rest and during stress, of fasting values of TSH and growth hormone, thyroxine, testosterone, and of urinary skin collagen metabolites and urinary tryptamine. Information from articles reviewed here is organized under the following headings: Problems studied, Methods of investigation and results, Conclusions, Summary of results, Discrepancies, Areas of research needed, and Practical clinical implications. Highlights of pertinent data contained in the original articles are organized in tables to facilitate direct comparisons between similar studies and between data on healthy and subjects with spinal cord injuries. PMID- 6293406 TI - Peripheral motor neuropathy caused by excessive intake of dapsone (Avlosulfon). AB - A case of selective peripheral motor polyneuropathy caused by excessive intake of dapsone is described. The condition was characterized by a peripheral muscle weakness in all limbs, normal sensitivity and present, although weak, muscle reflexes. Neurophysiologically, low amplitude muscle responses, prolonged distal latencies and reduced motor conduction velocities were found together with electromyographic signs of denervation. Sensory neurography was normal. The patient showed a complete clinical recovery and a marked neurophysiological restitution after termination of the excessive drug intake. The patient was found to acetylate dapsone at a slow rate. The case is compared with those previously reported in the literature. PMID- 6293407 TI - Biological effect of D,L-ureidosuccinic acid dihydrazide on Pseudomonas pseudomallei and Salmonella typhimurium. PMID- 6293408 TI - Interaction of diet and toxicity--the future role of purified diet in toxicological research. AB - There is a growing awareness of the importance of diet as a determinant of the toxicity of many compounds. This paper briefly reviews some of the ways in which diet affects toxicity, and draws some conclusions regarding the ideal diet for toxicological investigations. Then the two competing types of diet, stock and purified (including those frequently called semi-synthetic and semi-purified), are described and their characteristic strengths and weaknesses are discussed. Stock diets are very variable commodities, may be nutritionally poorly balanced, and also contain many non-nutritive components that influence toxicity. The formulation and preparation of purified diet are discussed. It is concluded that investigations of the mechanisms of toxicity, as well as studies of absorption, distribution and metabolism of toxic compounds, could benefit from the use of well-defined purified diets. PMID- 6293409 TI - The isolation and partial characterisation of a cytomegalovirus from the brown rat, Rattus norvegicus. AB - This report describes the isolation and partial characterisation of a cytomegalovirus (CMV) from the wild brown rat (Rattus norvegicus). The isolate was sensitive to ether and heat treatment at 56 degrees C/30 minutes, and had a characteristic herpes virus morphology. In rat embryo fibroblast cells, new virus was detectable by 18 hours after inoculation. The cytopathic effect consisted of a focal rounding of cells which developed to involve the entire monolayer. Inoculation of newborn rats produced mortality of 67-75 per cent which was reduced to 33-46 per cent by intra-peritoneal inoculation or tissue culture passage. Highest virus concentrations were found in the liver and spleen 14 days after inoculation. The virus was also pathogenic (to a lesser extent) to newborn BALB/c mice. It was related to the Osborn strain of mouse CMV by fluorescent antibody testing but distinct by the serum-neutralisation test. PMID- 6293410 TI - Comparative immunogenicity of 146S, 75S and 12S particles of foot-and-mouth disease virus. PMID- 6293411 TI - Differentiation of pseudorabies (Aujeszky's disease) virus strains by restriction endonuclease analysis. PMID- 6293412 TI - Enteroviral conjunctivitis and its neurological complications. PMID- 6293413 TI - Serotyping and subgrouping of rotavirus strains by the ELISA test. AB - A method is described for the serotyping and subgrouping of rotaviruses by enzyme linked immuno-sorbent assay (ELISA). For the isolates tested "blind" under code in parallel the serotyping results obtained by ELISA and serum neutralization of fluorescent focus formation were the same. After absorption of the typing antisera with purified heterotypic rotavirus, up to 128-fold differences in titres between isolates were observed in the ELISA test. The results of serotyping and subgrouping, by ELISA, of strains previously described and characterized by the authors' and other laboratories are also given. An attempt is made to correlate the serological findings with variations between virus strains based upon patterns of genome molecular weights as revealed by polyacrylamide gel electrophoresis. The application of this ELISA technique to epidemiological studies and vaccine research is discussed. PMID- 6293414 TI - Isolation of a cytomegalovirus-like agent from wild rats. AB - In 8 of 10 wild rats trapped in The Netherlands, an infectious viruslike agent was isolated predominantly from the salivary glands and could be serially passed in laboratory rats. In rat embryo cells a typical cytomegalo-like cytopathic effect was produced. The morphologic and cultural characteristics of the isolated agent were comparable with those of the mouse cytomegalovirus (MCMV). The virus nucleocapsid had a size of 92 nm and was not ether-resistant. The extracellular nucleocapsids were often enclosed by an outer layer of very variable shape and size. The formation of Fc receptors on cells infected with the rat virus could be demonstrated. The wild rats possessed neutralizing antibodies to the isolated agent. The rat agent grew only in rat embryo fibroblast cells while MCMV grew in rat and mouse embryo cells. The rat agent gave plaques in REF monolayers. Electron microscope studies showed the presence of nucleocapsids in the nucleus. PMID- 6293415 TI - Oncogenity of BK virus for immunosuppressed hamsters. AB - Tumors were induced by BK virus (BKV) inoculated intravenously in 3-week-old Syrian golden hamsters immunosuppressed with anti-lymphocyte serum or methylprednisolone acetate alone or in association with gamma-radiation (60Co). The induced neoplasms were ependymoma, carcinoma of pancreatic islets, lymphoma, osteosarcoma, undifferentiated sarcoma, kidney and renal pelvis carcinoma, pheochromocytoma and hemangiosarcoma. High levels of insulin and glucagon and altered concentrations of glucose were detected in blood of animals with tumors of pancreatic islets. No antibodies to BKV tumor antigen (TAg) and low levels of hemagglutination-inhibition antibodies to BKV viral coat protein Ag were detected in hamster sera. BKV TAg was found in tumors by complement fixation. Blot hybridization analysis of tumor DNA showed the presence of both free and integrated BKV genomes in tumor cells. BKV DNA inoculated intravenously and subcutaneously in immunosuppressed or immunocompetent hamsters was not oncogenic, whereas it was weakly oncogenic when inoculated intracerebrally. PMID- 6293416 TI - Sensitivity of bovid herpesvirus 2 replication to temperatures found in the natural host. AB - Two isolates of bovid herpesvirus 2 replicated poorly in bovine testicular cells and fetal kidney cells at 39--40 degrees C, temperatures commonly observed in virus-infected cattle. High viral titers occurred in replicate cultures at 30--37 degrees C. Persistent viral infections were noted in cultures maintained at 40 degrees C. Interferon-like activity was not responsible for the high-temperature restriction since the level was not significantly different between virus infected cultures incubated at 35 degrees or 40 degrees C. Spontaneous viral inactivation was only three times as rapid at 41 degrees as at 35 degrees C. Analysis of temperature shift experiments with respect to results of growth studies and electron microscopy indicated a temperature sensitive event late in replication. The sensitivity of bovid herpesvirus 2 replication to elevated temperature provides a possible explanation for the extensive viral growth and occasionally severe lesions which are limited exclusively to the skin of infected cattle. PMID- 6293417 TI - Infection of a calf with the enteric coronavirus strain Paris. AB - A tissue-culture-grown enteric coronavirus infected the whole of the gastrointestinal tract and caused enteritis and diarrhoea in a 10-day-old gnotobiotic Friesian calf. Diarrhoea occurred 2 days after inoculation and excretion of virus in faeces and rectal swabs increased until necropsy at 3 days. Virus growth, as detected by indirect immunofluorescence, virus isolation, and thin section electron microscopy, was most extensive in the epithelium of the colon and rectum at 3 days after inoculation. No virus was detected in other organs or in the pleural, peritoneal and oropharyngeal cavities. The origin and host specificity of this coronavirus is discussed. PMID- 6293418 TI - Synthesis and methylation of ribosomal RNA in HeLa cells infected with the herpes virus pseudorabies virus. AB - The effects of infection with the herpes virus pseudorabies virus on the metabolism of HeLa cell ribosomal RNA were examined. There is a decline both in the synthesis of nucleolar 45S ribosomal precursor RNA and in its processing to mature cytoplasmic RNA. The methylated oligonucleotides in the ribosomal RNA species were studied. The methylation of cytoplasmic ribosomal RNA was essentially unchanged. However there was some undermethylation of the nucleolar precursor. If undermethylated RNA does not mature then this may partly explain the reduced processing in the infected cells. PMID- 6293419 TI - A DNA-nicking activity associated with the nucleocapsid of human cytomegalovirus. AB - Purified human cytomegalovirus nucleocapsids are able to transform supercoiled plasmid DNA into nicked circular DNA. The activity exhibits a ionic strength optimum of 0.15 M; has a broad pH and temperature dependence and is enhanced after pre-incubation of the nucleocapsids with NP40. PMID- 6293420 TI - [Effects of pancreatic kallikrein on cyclic nucleotide levels of murine lymphocytes and antigen or mitogen induced lymphocyte proliferation]. PMID- 6293421 TI - Chemotherapy in the management of small cell lung cancer. PMID- 6293422 TI - [Pulmonary pneumocystosis]. AB - Sixteen autopsy cases of adults and older children who had contracted pneumocystosis of the lung were observed macro- and microscopically. Electron microscopical observation was done in 5 of these 16 cases. Also experimental studies on the pathogenesis of this disease were carried out on 20 rats. Most of the autopsy cases in this study had hematological neoplasia such as leukemia and malignant lymphoma which had been treated with cortisone and cytostatics. We found that intraalveolar foamy substances in this disease chiefly consisted of thin- and thick-walled Pneumocystis carinii which had many pseudopodias on the surface. Through these pseudopodias, pneumocystis were bound to each other and to the alveolar wall. In many cases desquamation of alveolar epithelial cells, formation of hyaline membrane and organization of alveolar contents were seen. Association with cytomegalovirus pneumonia was found in 12 cases. PMID- 6293423 TI - [Cancer of the stomach arising from chronic ulcers (histological and histogenetic problems]. AB - Malignization of chronic peptic ulcers occurs in 9.7% of cases. Carcinomas of various histological structures develop in the edges of chronic ulcers: dark-cell adenocarcinomas, low-differentiated, colloid cancers, and combinations thereof. Dark-cell adenocarcinomas are most prevalent. In the early stages of development all carcinomas are detected in the upper parts of the mucosa. The basic processes developing in the surrounding mucosa are all of the same type (proliferation of dark cells of the pit-cervical parts of the glands, intestinal metaplasia, atrophic processes). The current opinion held in the literature that various histological variants of gastric carcinoma are determined by different cell sources seems to be erroneous. All or at least the majority of gastric carcinoma due to chronic ulcer develop from the cells of the fossa-cervical parts. PMID- 6293424 TI - [Characteristics of regional lymph nodes in breast cancer (quantitative histochemical study)]. AB - The changes in axillary lymph nodes in mammary gland carcinoma of different histological types, metastasizing and nonmetastasizing, as well as after radiation therapy and in fibroadenomatosis were studied. The study was carried out on cryostate sections by histological and histochemical methods. Signs of activation of lymph nodes were clearly seen only in solid carcinoma, not always manifested in adenocarcinomas and scirrhous carcinomas, and undetectable in fibroadenomatosis. The quantitative determination of enzymes and nucleic acids showed differences in their activity between fibroadenomatosis and carcinomas. Proliferation processes dominated significantly over lymphocyte differentiation in carcinoma, increasing even more in metastasizing tumors. Pre-operative irradiation did not inhibit metabolism or proliferative activity of the cells. PMID- 6293426 TI - [Angiofibroma of the small intestine]. PMID- 6293425 TI - [Role of perifocal changes in lung cancer and its cytologic diagnosis]. PMID- 6293427 TI - [Malignant fibrous histiocytoma]. PMID- 6293428 TI - Acute encephalopathy in twins due to adenovirus type 7 infection. PMID- 6293429 TI - Branch artery occlusion. An unusual complication of external carotid embolization. AB - A case of retinal branch artery occlusion was caused by migration of emboli, presumably via collateral circulation, during therapeutic embolization of the maxillary artery. Migration of particles to the ophthalmic circulation is unusual with embolization of the branches of the external carotid artery. Meticulous technique, careful angiographic monitoring, and proper selection of embolic material may reduce, but not eliminate, migration of emboli to undesirable locations. Therapeutic embolization of vascular tumors and malformations in the external carotid territory is a recent radiologic innovation that is becoming increasingly popular. Therefore, we may expect to see more ocular complications from aberrant emboli as the use of this technique becomes more widespread. PMID- 6293430 TI - Adenoid cystic carcinoma. PMID- 6293431 TI - Angiofibroma of the maxillary antrum. PMID- 6293432 TI - [Stereomicroscopy of exfoliated epithelial cells]. PMID- 6293433 TI - Studies on Hofbauer cells in villous stroma of hydatidiform mole. PMID- 6293434 TI - Metabolic properties of mouse uterine endometrial cells after isolated with collagenase. AB - Uterine horns from unmated, pseudopregnant (days 1-5) and pregnant (days 4 and 5) mice were dissected and treated with collagenase (5 mg/ml) in a Ringer solution to isolate viable endometrial cells with maximal recovery of the luminal epithelium. The oxidative metabolism of the cells incubated with radioactive glucose significantly increased on day 4 of pregnancy, was stimulated during pseudopregnancy by the addition of either concanavalin A (Con A, 100 micrograms) or insulin (0.1 i.u.), and responded to a variety of traditionally used metabolic inhibitors in a manner consistent with that of cells possessing normal transport and oxidative functions. The metabolism of endogenous materials was an important feature of the cells and no evidence of a Crabtree effect was detected in the presence of various exogenous substrates. The cells also failed to alter their capacity to oxidize radioactive glucose after a 3-h pre-incubation period in vitro. Although the cells agglutinated in the presence of Con A at all reproductive stages studied, the agglutination indices significantly increased both on day 1 and on days 4 and 5 post coitum. The results indicated that metabolic changes and membrane modifications occurring in the cells at the time of implantation may have important roles in facilitating the induction of the decidual cell reaction. It was concluded that, if further technological improvements of the collagenase procedure succeed in producing pure epithelial cell preparations, the cells should be suitable for use in studies designed to elucidate these roles. PMID- 6293435 TI - Satellite DNA sequences in the red kangaroo (Macropus rufus). AB - There is a complex pattern of satellite DNA sequences in M. rufus which are revealed by addition of Ag+ or dye (Hoechst 33258) to the DNA ink Cs2SO4 or CsCl equilibrium density gradients. Six satellite DNA fractions have been isolated; these have buoyant densities in neutral CsCl of 1.692, 1.704, 1.705, 1.707 (two), 1.710 and 1.712 g/ml compared with 1.696 g/ml for the main band DNA. Each satellite accounts for 1-3% of the DNA of the genome. The satellites are located in the centromeric heterochromatin of the chromosomes, in the nucleolar organizer region and in interstitial bands on some of the autosomes, each satellite having a unique distribution. Nucleic acid hybridization showed that six of the satellite sequences are also present in the genomes of the wallaroo and the red necked wallaby, with sequence divergences of only 1-2% relative to the sequences in the red kangaroo. PMID- 6293436 TI - Comparison of restriction endonuclease profiles of DNA from local herpes simplex virus type 2 (HSV-2) strains. AB - Comparisons of restriction endonuclease profiles of DNA isolated from nine genital HSV-2 strains isolated from 1973 to 1981 were made using six restriction endonucleases (Bgl II, Hind III, Xba 1, Eco RI, Bam HI and Hpa I). The only variation detected in the DNA profiles was found with Eco RI and Bam HI, and in each case all the isolates could be classified into two groups. The difference between the two Eco RI groups could be accounted for by the absence of an Eco RI cleavage site from the HSV DNA. However, if comparison of the profiles of Eco RI and Bam HI cleaved DNA of the individual HSV-2 strains is made then this yields four different groups. The complete Bgl II, Xba I, Eco RI and Hpa I restriction endonuclease maps for the DNA from one isolate (HSV-2M1) are shown. Little variation in the DNA profiles can be found in local genital HSV-2 strains isolated over a 9-year period, when individual restriction enzymes are used. Thus, restriction endonuclease analysis has limited use in local epidemiological studies of genital HSV-2 isolates unless a wide variety of restriction endonucleases are used. However, the technique offers rapid differentiation between HSV-2 and HSV-1 local strains by using Hpa I endonuclease cleavage of HSV DNA. PMID- 6293437 TI - Bluetongue virus serotype 20 infection in pregnant Merino sheep. AB - Eleven maiden Merino ewes, free of antibody to bluetongue virus serotype 20 (BTV 20) in agar gel immunodiffusion and serum neutralisation tests, were mated once with a ram. Ten ewes were inoculated with BTV-20 35 to 42 days after service, and one ewe was left as an uninoculated control. One of the inoculated ewes and the control ewe remained uninfected throughout the experiment. Eight of the remaining 9 ewes showed clinical signs ranging from mild to moderate, and the other showed no clinical signs of infection. BTV-20 viraemia was detected in ewes between days 3 and 11 post inoculation, and the serum antibody response was followed. The control ewe and 5 of the 9 infected ewes were pregnant when examined 90 to 97 days after service. Each of these animals produced a normal lamb. There was no evidence of abortion in the remaining 5 ewes, and no transplacental transfer of virus was detected in the lambs of the 5 infected ewes. At necropsy, 46 days after the birth of the last lamb, no gross or microscopic lesions were observed in either the ewes or lambs. PMID- 6293438 TI - Studies on a vaccine against infections bursal disease. AB - An infectious bursal disease vaccine, registered for use in breeder flocks, was studied for efficacy on the day-old offspring of vaccinated hens and for virulence in susceptible day-old and 6-week-old chickens. When given to susceptible day-old chicks and 6-week-old cockerels, the vaccine was found to induce atrophy and pathology of the bursa of Fabricius similar to that observed in field infections. Chicks vaccinated at day-old had markedly lowered titres in the haemagglutination inhibition test to Newcastle disease virus, when this was given 2 weeks later, but the response of the 6-week-old cockerel was similar to that of control birds. Maternal antibody induced by the vaccine protected chicks against infection at day-old. PMID- 6293439 TI - Laboratory evaluation of a living attenuated vaccine against bluetongue type 20 virus. AB - Living attenuated bluetongue Type 20 virus vaccine was tested in 9 to 12 month old Australian Merino sheep, held in air conditioned, insect-free accommodation. The vaccine appeared avirulent and immunogenic and protected against infection with a second dose of homologous vaccine virus. No enhancement of virulence or significant change in immunogenicity was observed when the vaccine was passaged 3 times through sheep without antibody to bluetongue virus. PMID- 6293440 TI - Field trials of an immunization procedure against hemorrhagic enteritis of turkeys. AB - The efficacy of oral vaccination for hemorrhagic enteritis of turkeys was assessed by comparing flocks raised on the same premises, under the same management, with and without vaccination. The immunizing virus, a strain of marble spleen disease virus of pheasants, was administered via the drinking water. Vaccinated and unvaccinated turkeys differed significantly in feed conversion rates and spleen weights after challenge. PMID- 6293441 TI - Lesions induced in the respiratory tract of chickens by serologically different adenoviruses. AB - Ten strains of adenovirus representing 10 serotypes were administered intratracheally to two groups of 3-week-old specific-pathogen-free chickens. Birds in group 1 were given only adenovirus. Birds in group 2 were inoculated with a virulent infectious bursal disease virus (IBDV) by eye-drop at one day of age as well as with the adenovirus at 3 weeks. The chicks were examined daily. Respiratory rales were observed in some birds with the dual infection. Gross pathologic alterations were minimal and limited to multiple scattered, pale foci in the lungs of an occasional bird in various groups. Histopathologic changes in the lungs were those of multifocal interstitial and, occasionally, diffuse pneumonia. The pneumonic lesions were more severe in the chickens given IBDV. Tracheitis was seen with two of the serotypes. PMID- 6293442 TI - Preparing hemagglutinating antigen from isolates of infectious bronchitis virus. AB - The hemagglutinating (HA) activity of 14 strains of infectious bronchitis virus (IBV) was investigated. The optimal conditions for IBV antigen preparation include inoculation of 10- or 11-day-old specific pathogen-free embryonated eggs and incubation for 30 hours at 37 C. Embryos were inoculated via the allantoic cavity with 0.1 ml of a low embryonic passage of the virus (10(7) to 10(8) EID50/ml). Allantoic fluid was harvested and pooled, and a 100-fold concentration of virus particles was achieved by centrifugation for 3 hours at 30,000 x g. Virus pellets were resuspended in Tris-hydrochloride buffer containing 3 units of phospholipase-C (type-1) enzyme/ml and incubated for 2 hours at 37 C. All IBV strains tested demonstrated positive HA activity with chicken red blood cells. The antigen was stored in liquid state or lyophilized at 4 C. PMID- 6293443 TI - Evidence for bursal involvement in the pathogenesis of hemorrhagic enteritis of turkeys. AB - The pathogenesis of hemorrhagic enteritis in turkey poults infected with hemorrhagic enteritis virus (HEV) at 3 days or at 2 or 5 weeks of age was compared with pathogenesis in poults that had been chemically bursectomized neonatally and exposed to cell-culture-propagated virus at 2 or 5 weeks of age. Conventional poults exposed to HEV at 2 or 5 weeks developed clinical disease, and mortality ranged from 38% to 100%. In addition to the splenic and intestinal lesions usually seen with HEV infection, the pancreas, bursa of Fabricius, and thymus were also affected. In contrast, although they were free from detectable maternal antibody, poults infected with HEV at 3 days of age failed to develop clinical disease or mortality; however, virus was demonstrated by histological and electron microscopic examinations in spleens of these poults. Neonatal chemical bursectomy completely prevented the clinical signs, gross lesions, and mortality induced by HEV in poults at 2 or 5 weeks of age. These findings strongly suggest that an intact bursa is necessary for HEV to induce disease in turkeys. PMID- 6293444 TI - An in vitro assay for quantifying the virus of avian encephalomyelitis. AB - Chicken embryo brain (CEB) cell cultures support the replication of embryo adapted strains, vaccine strains, and field isolates of avian encephalomyelitis virus. A centrifugal force of 1,500 X g was applied during virus adsorption. Viral antigen was detected in the infected cells by using the indirect fluorescent-antibody technique (IFAT). Combining the infectivity of the virus in CEB cell culture with the ability to detect viral antigen by the IFAT resulted in the development of a virus-titration method. This in vitro assay proved to be more sensitive than the standard embryo-inoculation assay. It was concluded that the in vitro assay provides a satisfactory alternative to the embryo-inoculation assay. PMID- 6293445 TI - Pathologic effects of 2,2',4,4',5,5'-and 2,3',4,4',5,5'-hexabromobiphenyl in white leghorn cockerels. AB - Pathologic effects of 2,2',4,4',5,5'-hexabromobiphenyl (HBB), 2,3',4,4',5,5'-HBB, and a commercial mixture of polybrominated biphenyls (PBB) were compared in White leghorn cockerels. Diets containing 1, 10, or 100 ppm PBB, 4 or 10 ppm 2,3',4'4',5,5'-HBB, or 10 or 62 ppm 2,2',4,4',5,5'-HBB were fed for 28 days. Doses of 10 ppm of each chemical were used to provide a direct comparison of toxicity. Since nearly 4% of PBB consists of 2,3',4,4',5,5'-HBB and approximately 62% consists of 2,2',4,4',5,5'-HBB, effects of doses of 4 and 62 ppm, respectively, were compared with effects of 100 ppm of PBB to determine if either of the congeners were mainly responsible for the pathologic effects caused by the mixture. Liver weights were increased in cockerels fed diets containing 62 ppm of 2,2',4,4',5,5'-HBB or 10 or 100 ppm PBB. Hepatocytes were enlarged and vacuolated and lymphoid cells of the bursa of Fabricius were depleted by 10 ppm 2,3',4,4',5,5'-HBB, ppm 2,2',4,4',5,5'-HBB, and 10 or 100 ppm PBB. These dietary concentrations caused ultrastructural changes in hepatocytes consisting of vacuolation, increased smooth endoplasmic reticulum, swollen mitochondria, and disruption of mitochondrial cristae. When either of the congeners were given in concentrations relative to their concentrations in PBB, they were less toxic than the mixture. When concentrations in diets were equal, PBB caused more severe effects than 2,3',4,4',5,5'-HBB. The least effects were seen with 2,2',4,4',5,5' HBB. Results indicate that the two congeners chosen for study are not individually as toxic as the parent mixture. PMID- 6293446 TI - Pathogenicity of avian leukosis viruses. AB - Three methods were used in attempts to obtain non-oncogenic avian leukosis virus for possible use as an immunoprophylactic agent for the control of lymphoid leukosis in chickens. These were: 1) isolate a nononcogenic virus from commercial breeder flocks experiencing very little or no lymphoid leukosis; 2) obtain a non oncogenic recombinant from mixed infection of a strain with low oncogenicity, Rous-associated virus-60 (RAV-60), with RAV-1 or RAV-2 in cell culture; and 3) attempt to attenuate subgroup A avian leukosis virus by serial passage in avian cell culture. Of 43 isolates obtained from field sources, all were pathogenic except one, and its pathogenicity was questionable because of the low amount of virus tested. All 42 clones from mixed infection of highly oncogenic and poorly oncogenic virus and all clones passaged serially in cell culture were oncogenic. PMID- 6293447 TI - Susceptibility of ducks and duck-origin cell cultures to infectious bursal disease virus. AB - Specific-pathogen-free (SPF) ducks that were 1, 3, 4, 7, 10, 30, and 180 days old were inoculated experimentally orally or nasally with infectious bursal disease virus (IBDV). Attempts to induce clinical disease in ducks with strain J1 or FK 78 of IBDV were unsuccessful. Virus-recovery attempts from organ and intestinal contents were also unsuccessful. No significant gross or histopathological lesions were found in liver, spleen, kidney, heart, or bursa of Fabricius of 1- and 3-day-old ducks at 4 or 7 days postinoculation. The ratios of bursa weight to body weight of 1-, 10-, and 30-day-old inoculated and control ducks revealed no difference at 21 days postinoculation. The ducks responded serologically, however, by developing both virus-neutralizing and agar-gel-precipitin antibodies. Virus multiplied in embryonated duck eggs and duck embryo fibroblast cells but not in duck kidney cells. PMID- 6293448 TI - Heat inactivation of a duck adenovirus serologically indistinguishable from adenovirus 127. PMID- 6293449 TI - The effectiveness of selection for reduced avian leukosis virus shedding in different chicken strains. PMID- 6293450 TI - Pulmonary congestion and edema (marble spleen disease) of chickens produced by group II avian adenovirus. AB - "Marble spleen disease" of chickens was diagnosed in 22-week-old chickens. Total mortality was 8.9%. Deaths occurred over a period of 2 months. Gross lesions included pulmonary congestion, splenomegaly, hepatomegaly, and congestion of egg follicles. Microscopic lesions included pulmonary congestion and edema, and reticuloendothelial cell hyperplasia of the spleen with concurrent white-pulp necrosis and lymphocyte depletion. The pulmonary lesions were of sufficient intensity to have caused the death of fatally affected birds. Many of the hyperplastic reticuloendothelial cells contained basophilic intranuclear inclusions similar to those that characterize hemorrhagic enteritis of turkeys, marble spleen disease of pheasants, and adenovirus group II splenomegaly of chickens. These characteristic lesions, plus serologic identification of the causal virus, indicate that "marble spleen disease" caused by avian group II adenovirus was affecting the flock under study. This appears to be the first report of death of chickens due to pulmonary congestion and edema caused by spontaneous infection with avian group II adenovirus. PMID- 6293451 TI - Does subcuticular green polyglycolic acid suture tattoo? PMID- 6293452 TI - ACTH4-10 enhances retention of conditioned taste aversion learning in infant rats. PMID- 6293453 TI - Attenuation of the context effect and lack of unconditioned stimulus-preexposure effect in taste-aversion learning following treatment with DSP4, the selective noradrenaline neurotoxin. PMID- 6293454 TI - Oxidative phosphorylation function of two mitochondrial preparations from heart: effects of ischaemia and cytochrome C. PMID- 6293455 TI - Post-excitatory depression in thoracic sympathetic efferent neural traffic during a cardiogenic hypertensive chemoreflex. AB - Serotonin injected in the left atrium activates a cardiogenic hypertensive chemoreflex in dogs. To elucidate patterns of the neural traffic, records were obtained from thoracic sympathetic efferent nerves (either the anterior ansa of the left stellate ganglion or the T4 input to the left stellate) in 8 anesthetized dogs with chest open. Serotonin (200 micrograms, left atrium) caused a massive sympathetic discharge during the hypertension and bracardia characteristic of the chemoreflex. Following the initial sympathetic discharge, there was a consistent post-excitatory depression of neural traffic, to a level significantly less than control discharge (two-tailed p less than .05). This post excitatory depression began 11 +/- 5.4 (S.D.) seconds after injection of serotonin and 6.6 +/- 5.3 seconds after the peak neural discharge. It lasted 140 +/- 94 seconds, being maximal initially with gradual recovery. Complete block of the hypertension by the combined administration of phentolamine, propranolol, and nitroglycerin failed to abolish the efferent neural events, including post excitatory depression, in all but one dog. We conclude that post-excitatory depression in thoracic sympathetic efferent neural traffic cannot be mediated exclusively through the secondary engagement of a baroreceptor mechanism and that it most likely is an integral part of the cardiogenic hypertensive chemoreflex. PMID- 6293456 TI - [Protective action of undegraded and S-sulfonated 7S antibodies in 4 different experimental virus infections]. PMID- 6293457 TI - Nucleoside exchange catalysed by the cytoplasmic 5'-nucleotidase. AB - The inhibition of the cytoplasmic 5'-nucleotidase (EC 3.1.3.5) by its product, inosine, was studied with a partially purified preparation of the enzyme from rat liver. Inhibition of Pi production was found to be due to exchange of the inosine moiety between inosine and IMP. Exchange was not catalysed by reversal of the hydrolytic reaction, suggesting, instead, the mediation of an enzyme-phosphate intermediate. Two models for the catalytic mechanism are proposed and rate equations for the dependence of Pi production on inosine concentration are derived. The experimentally determined dependence was consistent with a mechanism in which hydrolysis of the enzyme-phosphate intermediate occurred only when it was unoccupied by inosine. This conclusion suggests that inosine analogues that cannot participate in exchange should inhibit the enzyme. Such inhibitors might be useful in defining the enzyme's physiological role or as pharmacological agents to decrease breakdown of purine nucleotides. The possibility that nucleoside exchange provides an alternative route for the phosphorylation of mutagenic or cytotoxic nucleoside analogues should also be considered. PMID- 6293458 TI - Identification and purification of a liver microsomal glucose 6-phosphatase. AB - 1. Hepatic glucose 6-phosphatase activity was purified 65-fold in good yield over that in cholate-solubilized microsomal fractions. 2. This preparation still contained five major polypeptides and numerous minor contaminants. 3. The smallest of the five major polypeptides (Mr approx. 18 500) could be purified from heat-treated microsomal fractions. 4. Antisera raised against the heat stable protein doublet was used to immunoprecipitate specifically glucose 6 phosphatase activity from cholate-solubilized microsomal fractions. 5. This work indicates that hepatic microsomal glucose 6-phosphatase appears to be one or both of the low-molecular-weight heat-stable polypeptides. PMID- 6293459 TI - Superoxide-forming NADPH oxidase preparation of pig polymorphonuclear leucocyte. AB - A phagocytic vesicle fraction with high NADPH-dependent superoxide-forming activity was obtained in large quantity from pig blood polymorphonuclear leucocytes, phagocytosing oil droplets in the presence of cyanide. The activity of the homogenate of the phagocytosing cells was 40 times that of the resting cells, and 70% of the activity in the homogenate was recovered in the phagocytic vesicle fraction. Essentially all of the superoxide-forming activity was extracted by repeated extraction with a mixture containing deoxycholate and Tween 20. The extract had a superoxide-forming activity of 1 mumol/min per mg of protein with NADPH, and one-fifth of this with NADH, Km values being similar to those of the vesicle fraction (40 microM for NADPH and 400 microM for NADH). A stoichiometric relationship of 1:2 for NADPH oxidation and superoxide formation was obtained, in agreement with the reaction NADPH +2O2 leads to NADP+ + 2O2 -. + H+. The activity of the extract was enhanced 2-fold by the addition of FAD, suggesting that the flavin is a component of the enzyme system. The Km value for FAD was 0.077 microM. The activities in both vesicle fraction and extract were labile even on refrigeration, but could be kept for several months at -70 degrees C. PMID- 6293460 TI - Occupancy of phosphorylation sites in pyruvate dehydrogenase phosphate complex in rat heart in vivo. Relation to proportion of inactive complex and rate of re activation by phosphatase. AB - The [gamma-32P]ATP-back-titration method of estimating occupancy in vivo of the three phosphorylation sites in the pyruvate dehydrogenase complex was improved in precision by specific analysis with trypsin/formic acid, by more effective prevention of site-2 dephosphorylation during purification with NaF, and by other refinements. Disproportionation of phosphorylated complexes during purification was excluded. With this improved method it was shown that the relationship between occupancy of sites and the proportion of complex in the inactive form in rat heart in vivo is closely similar to that measured directly in heart mitochondria by incorporation of [32P]Pi. In the heart in vivo (as in mitochondria), occupancy of site 1 correlated linearly with the proportion of inactive complex. Occupancy of sites 2 and 3 only approached equivalence to that of site 1 when 99% of the complex was inactive (starved or diabetic rats). When 70% or less of the complex was inactive (resting or exercising fed normal rats), occupancy of sites 2 and 3 was minimal (3 less than 2) relative to site 1. The initial rate of re-activation by phosphatase of phosphorylated complex from hearts of resting or exercising fed normal rats was approximately three times that of complex from 48 h-starved rats. PMID- 6293461 TI - Protein turnover and proliferation. Failure of SV-3T3 cells to increase lysosomal proteinases, increase protein degradation and cease net protein accumulation. AB - The contrasting control of lysosomal proteinases, protein turnover and proliferation was studied in 3T3 and SV-3T3 (SV-40-virus-transformed 3T3) cells. 1. In 3T3 cells, net protein accumulation proceeded from 5%/h (doubling time, T(d)=14h) in growing cells to 0%/h as cells became quiescent. SV-3T3 cells never ceased to gain protein, but rather decreased their protein accumulation rate from 6-7%/h (T(d)=10-12h) to 2%/h (T(d)=35-40h) just before culture death in unchanged medium. 2. In both cell types the rates of protein synthesis per unit of protein (a) were proportional to the initial serum concentration from 0 to 6%, and (b) declined under progressive depletion of undefined serum growth factors. In depleted growth medium, leucine incorporation per unit of protein in 3T3 and SV 3T3 cells declined to almost equal synthetic rates while the 3T3 cell existed in a steady state of zero net gain, and the SV-3T3 cell continued to gain protein at a rate of 2%/h. 3. Whereas a large fraction of the control of 3T3-cell net protein accumulation can be accounted for by an increase in degradation from 1%/h to 3%/h, the SV-3T3 cell did not exhibit a growth-related increase in degradation appreciably above 1%/h. 4. Thus, by using first-order kinetics, the continued net protein accumulation of the transformed cell can be accounted for by a failure to increase protein degradation, whereas fractional synthesis can be made to decline to a rate similar to that in the quiescent non-transformed cell. 5. Upon acute serum deprivation, both cell types similarly exhibited small rapid increases in proteolysis independent of cell growth state or lysosomal enzyme status. 6. The 3T3 cell increased its lysosomal proteinase activity in conjunction with increase in the growth-state-dependent proteolytic mechanism; however, the SV-3T3 cell failed to increase lysosomal proteinases or the growth-state-dependent proteolytic mechanism. PMID- 6293462 TI - Respiration of the rumen ciliate Dasytricha ruminantium Schuberg. AB - The endogenous respiration of the rumen ciliate Dasytricha ruminantium maintained under an O2 tension of 2kPa (approximately 0.02 atm) was partially inhibited by KCN (40% inhibition) and NaN3 (58% inhibition). The organisms lack cytochromes, and sensitivity of respiration to KCN, NaN3, chloroquine and quercetin suggest that the operation of flavoprotein-iron-sulphur-mediated electron transport. As in Tritrichomonas foetus, hydrogenosomal respiration can be stimulated by the addition of CoA in the presence of 0.025% Triton X-100; stimulation by ADP was not detected. Stimulation of pyruvate-supported O2 uptake by Pi suggests that acetate is produced via acetyl phosphate. PMID- 6293463 TI - The relationship between collagen and C1q biosynthesis in cultured human fibroblasts. AB - The relationship between collagen and C1q biosynthesis has been investigated in cultured human fibroblasts. This was done by measuring the effects of variation in cell density, inhibition of prolyl hydroxylase and complexing of C1q on synthesis and/or secretion of both proteins. It was found that synthesis and secretion of both proteins were not co-ordinate, and that therefore regulation of expression of both proteins is probably not linked. However complexing of C1q did result in marked stimulation in collagen synthesis, suggesting that the fibrosis which follows inflammation may result from binding of C1q to the immune complexes formed in the inflammatory process. PMID- 6293464 TI - Processing of calcitonin and epidermal growth factor after binding to receptors in human breast cancer cells (T 47D). AB - 125I-labelled calcitonin and 125I-labelled epidermal growth factor (EGF) bound to T 47D breast cancer cells at 37 degrees C in a manner that became increasingly resistant to removal by acid pH. Bound 125I-labelled EGF became resistant to acid removal more rapidly than did bound 125I-labelled calcitonin. The shift from acid accessibility to acid inaccessibility was energy-dependent since it was not seen at 4 degrees C and was inhibited in the presence of cell metabolic inhibitors. Radioactivity removed by acid represented intact hormone as assessed by trichloroacetic acid precipitation, whereas radioactivity released spontaneously by the cells was trichloroacetic acid-soluble. Inclusion of 10 mM-NH4Cl in the incubation medium resulted in an accumulation of cell-associated radioactivity without affecting the shift to acid inaccessibility. The accumulated radioactivity was relatively more trichloroacetic acid-precipitable in comparison with that associated with control cells. These data are consistent with internalization of receptor-bound EGF and a similar though slower mechanism of processing for receptor-bound calcitonin. The predominant route of hormone release from cells seems to occur via intracellular degradation rather than dissociation from cell-surface receptors. PMID- 6293465 TI - Dynamics of carbon monoxide recombination to fully reduced cytochrome c oxidase in plant mitochondria after low-temperature flash photolysis. AB - Rebinding of CO to reduced cytochrome c oxidase in plant mitochondria has been monitored optically at 590-630 nm after flash photolysis at low temperature from 160 to 200 K. (1) Under 100%-CO saturation, CO rebinding exhibits a four-step mechanism. The thermodynamic parameters of the first phase have been determined; its activation energy, Ea1, is 38.9 kJ.mol-1 and its enthalpy, delta H+/-1, and entropy, delta S+/-1, of activation are respectively 37.5 kJ.mol-1 and -75.8J.mol 1.K-1. (2) When the CO concentration is decreased to 0.2%, rebinding still occurs according to a four-step mechanism. The rate constant of the first phase is CO concentration-independent. Under non-saturating conditions there is only one CO molecule per occupied site. The rebinding mechanism does not require additional CO molecules to be present in the haem pocket. (3) Dual-wavelength scanning experiments failed to detect optical forms correlated with the resolved phases. (4) Results are discussed with respect to previous work related to CO rebinding to mammalian cytochrome c oxidase and myoglobin. PMID- 6293466 TI - Proton translocation by the mitochondrial cytochrome b-c1 complex is inhibited by NN'-dicyclohexylcarbodi-imide. AB - NN'-Dicyclohexylcarbodi-imide at low concentrations decreases the H+/2e ratio for rat liver mitochondria over the span succinate to oxygen from 5.9 +/- 0.3 (mean +/- S.E.M.) to 4.0 +/- 0.1 and for the cytochrome b-c1 complex from 3.8 +/- 0.2 to 1.9 +/- 0.1, but has little effect on the H+/2e ratio of cytochrome oxidase. The decrease in stoicheiometry is due, not to uncoupling or inhibition of electron transport, but to inhibition of proton translocation. NN' Dicyclohexylcarbodi-imide thus 'decouples' proton translocation in the cytochrome b-c1 complex. PMID- 6293467 TI - Regulation of proline 3-hydroxylation and prolyl 3-hydroxylase and 4-hydroxylase activities in transformed cells. AB - Prolyl 3-hydroxylase activity and the extent of collagen proline 3-hydroxylation were studied in six transformed and three control human cell lines. In the transformed cell lines, the enzyme activity was markedly high in two, similar to that in control cells in two and significantly low in two. The extent of proline 3-hydroxylation was markedly high in cell lines with high enzyme activity, but it was also significantly high in some transformed cell lines with enzyme activities similar to those in the controls. The results thus suggest that, in addition to the amount of enzyme activity present, the rate of collagen synthesis also affects the extent of proline 3-hydroxylation in the newly synthesized collagen. The effect of acute cell transformation on prolyl 3-hydroxylase and 4-hydroxylase activities was studied by infecting chick-embryo fibroblasts with Rous sarcoma virus mutant NY68, temperature-sensitive for transformation. At the permissive temperature prolyl 3-hydroxylase activity showed a more rapid increase and decrease than did prolyl 4-hydroxylase activity, the maximal activity for both enzymes being about 2.5 times that in the control chick fibroblasts. When the transformed cells were shifted to the non-permissive temperature the decays in the elevated enzyme activities were similar, suggesting identical half-lives. PMID- 6293468 TI - Tetracyano-2,2-bipyridineiron(iii), an improved electron acceptor for the spectrophotometric assay of beta-oxidation and of succinate dehydrogenase in intact mitochondria. AB - A recently described direct reading assay for beta-oxidation and for succinate oxidation in intact mitochondria using [Fe(CN)6]3- as final electron acceptor [Osmundsen & Bremer (1977) Biochem. J. 164. 621--633] has been improved by using instead tetracyano-2,2-bipyridineiron(III) [Fe(CN)4(bpy)]-, which gives a 2.6 times greater absorbance change on reduction. Some physical and kinetic properties of [Fe(CN)4(bpy)]- are described. The use of exogenous cytochrome c(III) as electron acceptor was also tested; this gives the largest absorbance change, although the absolute rate of reaction is only approx. one-third of that using [Fe(CN)6]3- or [Fe(CN)4(bpy)]-. PMID- 6293469 TI - Superoxide-dependent formation of hydroxyl radicals and lipid peroxidation in the presence of iron salts. Detection of 'catalytic' iron and anti-oxidant activity in extracellular fluids. AB - Synovial fluid from rheumatoid patients and normal cerebrospinal fluid contains micromolar concentrations of non-protein-bound iron salts that can promote lipid peroxidation and also the superoxide-dependent formation of hydroxyl radicals from hydrogen peroxide. These iron catalysts of oxygen radical reactions cannot be detected by conventional assays unless interfering high-molecular-weight substances, probably proteins, are removed by ultrafiltration or inactivated by exposure to low pH values. The bleomycin assay for ;catalytic' iron [Gutteridge, Rowley & Halliwell (1981) Biochem. J.199, 263-265] does not suffer from these artifacts. PMID- 6293470 TI - Sendai virus causes a rise in intracellular free Ca2+ before cell fusion. AB - 1. Sendai virus caused a large increase in the concentration of free Ca(2+) within human erythrocyte ghosts detected by the Ca(2+)-activated photoprotein obelin. 2. The increase in intracellular [Ca(2+)] preceded fusion. However, fusion could also be observed in the absence of a detectable rise in intracellular free [Ca(2+)]. 3. It was concluded that the increase in intracellular free [Ca(2+)] was not an absolute requirement for cell fusion, but may be necessary to produce fusion at the maximum rate. PMID- 6293471 TI - Proteolytic activation can produce a phosphatidylinositol phosphodiesterase highly sensitive to Ca2+. AB - The phosphatidylinositol phosphodiesterase of rat brain shows little activity under conditions likely to pertain in vivo (neutral pH and micromolar Ca(2+) concentrations). A short incubation of a brain supernatant with trypsin, or a longer pre-incubation of the supernatant alone, produce new forms of the enzyme, which are active under such conditions. A possible role of receptor-linked proteinases in initiating phosphatidylinositol catabolism is discussed. PMID- 6293472 TI - Studies on alpha-glycohydrolases (alpha-L-fucosidase, alpha-D-galactosidase, and alpha-D-mannosidase) in sera of diabetic patients and controls. PMID- 6293473 TI - The effect of glucose 2-deoxy-D-glucose and insulin on estradiol secretion by cultured human trophoblast. PMID- 6293474 TI - Specific binding of [3H]nitrendipine to membranes from coronary arteries and heart in relation to pharmacological effects. Paradoxical stimulation by diltiazem. PMID- 6293475 TI - Epinephrine-mediated stimulation of glucose uptake and lactate release by the perfused rat heart. Evidence for alpha- and beta-adrenergic mechanisms. PMID- 6293477 TI - Cis-diamminedichloroplatinum (II) modification of SV40 DNA occurs preferentially in (G+C) rich regions: implications into the mechanism of action. PMID- 6293476 TI - Evidence for multiple molecular weight forms of the chick intestinal 1,25 dihydroxyvitamin D3 receptor. PMID- 6293478 TI - 5-Methylthioribose kinase activity in plants. PMID- 6293479 TI - Prostaglandin E1-stimulable cyclic AMP formation from rat gastric antral organ culture: lack of effect on gastrin secretion. PMID- 6293480 TI - Prolactin modifies the prostaglandin synthesis, prolactin binding and fluidity of mouse liver membranes. PMID- 6293481 TI - Presence of "Ra" and "P"-site receptors for adenosine coupled to adenylate cyclase in cultured vascular smooth muscle cells. PMID- 6293482 TI - The effect of fructose 2,6-bisphosphate on the reverse reaction kinetics of fructose 1,6-bisphosphatase from bovine liver. PMID- 6293483 TI - The effect of antiarthritic drugs and related compounds on the human neutrophil myeloperoxidase system. PMID- 6293484 TI - Long-term effect of 2-deoxy-2-fluoroglucose on maintenance in culture of the neonatal B cell of rat. PMID- 6293485 TI - Binding, internalization and intracellular processing of 125I-epidermal growth factor purified by isoelectric focusing. PMID- 6293486 TI - Selective antilipolytic effect of bacitracin in the isolated fat cell. PMID- 6293487 TI - NAD-glycohydrolase activity in Xenopus laevis oocytes and early embryos. PMID- 6293488 TI - Differences in superoxide production by nonmigrating and migrating human monocyte subpopulations. PMID- 6293490 TI - Differential effects of GTP and cations on binding of labeled dimeric and monomeric enkephalins to neuroblastoma-glioma cell delta opiate receptors. PMID- 6293489 TI - Role of carbohydrate in human chorionic gonadotropin: deglycosylation uncouples hormone-receptor complex and adenylate cyclase system. PMID- 6293491 TI - Isolation of brain Ca2+-calmodulin tubulin kinase containing calmodulin binding proteins. PMID- 6293492 TI - Characterization of the mammalian beta-2 receptor in 8 M urea. PMID- 6293493 TI - The ATPMg-dependent phosphatase is present in mammalian vascular smooth muscle. PMID- 6293494 TI - Stereochemistry of naturally-occurring 25-hydroxyvitamin D3-26,23 lactone as determined by radioligand binding analysis and high-performance liquid chromatography. PMID- 6293495 TI - Reversible inactivation of 3-hydroxy-3-methylglutaryl coenzyme A reductase: reductase kinase and mevalonate kinase are separate enzymes. PMID- 6293496 TI - Mechanism of action of benzo(a)pyrene and nicotine on hormone production by rat pituitary tumor cells. PMID- 6293497 TI - Protective effect of EGTA on rat gastric membranes enriched in (H+-K+)-ATPase. PMID- 6293498 TI - Diltiazem enhancement of [3H]nitrendipine binding to calcium channel associated drug receptor sites in rat brain synaptosomes. PMID- 6293499 TI - EPR evidence for nickel-substrate interaction in carbon monoxide dehydrogenase from Clostridium thermoaceticum. PMID- 6293500 TI - Uncoupling by proteolysis of alpha-adrenergic receptor-mediated inhibition of adenylate cyclase in human platelets. PMID- 6293501 TI - ESR investigations at low temperatures of oxygen radical interaction with rat chromochelatin and metallothioneins. PMID- 6293502 TI - Nonenzymatic galactosylation of human LDL decreases its metabolism by human skin fibroblasts. PMID- 6293503 TI - Adenine nucleotide content of liver mitochondria increases after glucagon treatment of rats or isolated hepatocytes. PMID- 6293504 TI - A novel protamine kinase activity in human promyelocytic leukemia cells. PMID- 6293505 TI - Stimulatory effect of glucagon and dibutyryl-cAMP specifically on the Na+ independent amino acid transport of Chang liver cell. PMID- 6293506 TI - Selective activation of the converting enzyme inhibitor MK 421 and comparison of its active diacid form with captopril in different tissues of the rat. PMID- 6293507 TI - Relationship between cyclic AMP-dependent protein kinase activation and Ca uptake increase of sarcoplasmic reticulum fraction of hog biliary muscles relaxed by cholecystokinin-C-terminal peptides. AB - In hog terminal bile duct cholecystokinin peptides caused an activation of cyclic AMP-dependent protein kinase (A-PK) with cyclic AMP, followed by increase in Ca uptake of sarcoplasmic reticulum fraction (SR-F). By contrast, papaverine showed no activation of A-PK-induced Ca uptake by SR-F with cyclic AMP. The Ca uptake by SR-F was dependent on ATP and Mg2+, but the component phosphorylated was not the phosphoenzyme intermediate in Ca2+-ATPase. The effect of Ca uptake was blocked by the inclusion of a protein inhibitor of A-PK. The correlation coefficient between cyclic AMP-dependent SR-F phosphorylation and stimulated Ca uptake by the phosphorylated SR-F was 0.731 (P less than 0.01). These results suggest that one of the mechanisms by which CCK-4, CCK-8, and CCK-33 peptides relax isolated Oddi's sphincters of terminal bile ducts is activation of A-PK-induced Ca uptake by sarcoplasmic reticulum fraction and possibly also by plasma membrane. PMID- 6293508 TI - Effects of morphine on norepinephrine turnover in various functional regions of rat spinal cord. PMID- 6293509 TI - Uptake and retention of androgens by the rat ventral prostate and consideration of their use as site directing agents. AB - Elucidation of the mechanism whereby androgens are accumulated selectively by the prostate may help in the design of drugs for the treatment of prostatic cancer. The uptake and retention of [3H]testosterone, following intraperitoneal injection, by various tissues in the 24 hr castrate rat has been studied over an extended time course. The selectivity with which prostate, as compared with blood or other tissues, accumulated 3H was shown to be dose-dependent. At a low dose (0.15 microgram), selective prostatic accumulation was greater in 24 hr castrate and diethylstilboestrol-treated rats than in normal animals. Testosterone, 5 alpha-dihydrotestosterone and oestradiol, radioactively labelled, were each administered to 24 hr castrate rats by intraperitoneal injection. Specific prostatic accumulation of radioactivity was more dependent on steroid structure at a low dose than at a high dose (0.6 mg) and at the low dose (0.15 microgram) followed the order testosterone greater than 5 alpha-dihydrotestosterone greater than or equal to oestradiol. This order was surprising in view of the androgen receptor binding affinities of these steroids. It is concluded that small quantities of material could be directed with the greatest specificity to the prostate of castrate or diethylstilboestrol-treated animals if attached to testosterone. Androgens would be more useful for site-directed radiopharmaceuticals than cytotoxic agents. PMID- 6293510 TI - Selective inhibition of separated forms of cyclic nucleotide phosphodiesterase from rat heart by some cardio- or vaso-active butenolide derivatives. PMID- 6293511 TI - Inhibition of a high affinity cyclic AMP phosphodiesterase and relaxation of canine tracheal smooth muscle. AB - The cyclic nucleotide phosphodiesterase (PDE) activity of canine tracheal smooth muscle (CTS,) was examined. Column chromatography of soluble CTSM-PDE revealed five peaks of activity. One of these peaks (V) was examined further in this study and showed a high affinity for adenosine 3',5'-cyclic monophosphate (Km = 0.63 microM). Seven pharmacological PDE inhibitors were tested for their abilities to inhibit the peak V enzyme and also for their abilities to cause mechanical relaxation of CTSM strips in isolated tissue baths. A strong correlation (P greater than 0.001) between peak V PDE inhibition (-log Ki) and airway muscle relaxation (-log ED50) was found. PMID- 6293512 TI - Theophylline effect on the cyclic AMP degrading multienzyme sequence. AB - Membrane-bound 3'.5'-cyclic nucleotide phosphodiesterase (EC 3.1.4.17) is closely associated physically with nucleotidase and deaminase, thus forming an enzyme cluster of unique catalytic behaviour [H. Wombacher, Archs. Biochem. Biophys. 201, 8 (1980)]. This multienzyme cluster, which was found in the microsomal fraction of beef adrenal cortex, catalyses the degradation of cyclic AMP, via AMP and adenosine, to inosine. The present study shows how theophylline, a well-known inhibitor of the phosphodiesterase, acts on the membrane-bound multienzyme sequence. The findings were as follows. Firstly, as expected, theophylline inhibited the phosphodiesterase competitively. In particular, the high-affinity enzyme was inhibited by mM concentrations of theophylline. Phosphodiesterase activity was tentatively ascribed to two enzymes, one with a low Km [0.3 microM], one with a high Km [60 microM]. Secondly, theophylline inhibited the nucleotidase activity to a great extent. A detailed kinetic analysis showed the inhibition to be hyperbolic noncompetitive (alpha = 1, beta = 0.35 and Ki = 0.25 mM). Thirdly, theophylline did not inhibit the deaminase activity of the multienzyme sequence. A model of theophylline inhibition is suggested explaining how an effector could modulate the kinetic behaviour of an enzyme cluster by acting at a single allosteric site. Finally, in view of the existence of the cyclic AMP degrading multienzyme sequence and the effect of theophylline on it, the possibility is discussed that physiologically active adenosine is derived from cyclic AMP. PMID- 6293513 TI - Increased endothelial cell adherence, aggregation, and superoxide generation by neutrophils incubated in systemic lupus erythematosus and Felty's syndrome sera. AB - The ability of sera from patients with systemic lupus erythematosus (SLE) and Felty's syndrome to induce increased adhesiveness of normal human neutrophils (PMN) was investigated. PMN from normal healthy donors were incubated in sera from 19 patients with active SLE, 12 with inactive SLE, 20 with Felty's, 24 with rheumatoid arthritis, and 34 normal persons. After incubation, the degree of adherence of the PMN to human endothelial cells in culture, their aggregation, and superoxide (O2-) generation were determined. Sera from patients with both active SLE and Felty's syndrome induced significantly increased PMN adherence to endothelial cells and PMN aggregation in vitro, compared with normal sera. This increased adherence to endothelial cells was maintained after heat treatment (56 degrees C for 30 minutes) of the sera. In O2- generation experiments, sera from patients with active SLE induced significantly increased O2- release from normal PMN using both fresh and heat-treated sera. Sera from Felty's patients demonstrated the same effect with heat-treated sera but not ith fresh sera. When sera from patients with active SLE and Felty's syndrome were used, all three parameters correlated significantly with each other in individual patients. In contrast, sera from the 12 patients with inactive SLE and 24 rheumatoid arthritis patients without Felty's failed to induce significant differences in the three parameters studied when compared with 34 normal controls. Fractionation of 3 SLE sera and 1 Felty's serum on Sephadex G-200 demonstrated that the adherence enhancing factor was present in both IgG and IgG-excluded fractions. The observed increased adhesiveness of PMN induced by SLE and Felty's sera may, at least in part, contribute to the neutropenia which is common in these diseases. Increased O2- release associated with PMN adherence may contribute to endothelial cell damage and vascular injury, which is also a common manifestation of these diseases. PMID- 6293514 TI - [On the site of action of benzarone and some other agents influencing the intracellular concentration of cyclic nucleotides]. PMID- 6293515 TI - Further urinary metabolites of delta 1-tetrahydrocannabinol in man. AB - The in vivo metabolism of delta 1-tetrahydrocannabinol (delta 1-THC) was further investigated in man after oral administration. Five dicarboxylic acids were isolated from the urine and identified by gas chromatography-mass spectrometry. None of these acids have been identified in man before and two of them (3",4",5" trisnor-delta 1-THC-7,2"-dioic acid (15) and 5"-nor-delta 1-THC-7,4"-dioic acid (17)) have hitherto not been found in any species. The major dicarboxylic acid was 4",5"-bisnor-delta 1-THC-7,3"-dioic acid (16) corresponding to 8% of the excreted radioactivity in the urine. The other dicarboxylic acids (14, 18) were present in small amounts. PMID- 6293516 TI - Effects of bezafibrate on receptor-mediated and receptor-independent low density lipoprotein catabolism in type II hyperlipoproteinaemic subjects. AB - This study examines the effects of bezafibrate (200 mg t.i.d.) on LDL metabolism in 7 type II hyperlipoproteinaemic subjects. Eight weeks of treatment lowered plasma cholesterol and triglyceride by 10% and 30%, respectively (P less than 0.02). These reductions were associated with a fall in circulating VLDL (31%, P less than 0.02) and LDL (11%, P less than 0.05), while HDL cholesterol stayed the same. LDL metabolism changed during therapy. The plasma fractional clearance rate (FCR) of autologous [125I]LDL normalized from a low value of 0.256 +/- 0.048 (mean +/- SD) to 0.298 +/- 0.040 pools/day (P less than 0.001). This was attributable to a 65% increase (P less than 0.01) in receptor-mediated LDL catabolism since the clearance of simultaneously injected 1,2-cyclohexanedione modified [131I]LDL, which measures the receptor-independent pathway, was unaltered (FCR of [131I]cyclohexanedione/LDL in control phase = 0.194 +/- 0.030 pools/day; during drug treatment = 0.194 +/- 0.024 pools/day). We conclude that bezafibrate lowers plasma LDL in type II hyperlipoproteinaemia by promoting its degradation via high affinity receptors. PMID- 6293517 TI - Epidemiologic approaches to cancer etiology. PMID- 6293518 TI - Dissociation between normal hemispheres in delayed recognition of verbal and spatial cues of the same visual pattern. AB - Is each hemisphere capable of disentangling a specific kind of information from a complex 3-target set of stimuli in an expectancy-free delayed recognition experiment on normals? A tachistoscopical study set out to answer this question. It provided evidence for a double dissociation between the hemisphere of input of complex stimuli (comprising both spatial and verbal information) and the kind of information to be retrieved. PMID- 6293519 TI - Transmission and integration of biologically meaningful olfactory information after bilateral transection of the lateral olfactory tract in the rat. AB - The contribution of the olfactory inputs conveyed by the lateral olfactory tract (LOT) to the central olfactory areas in the onset of behavioral and electrophysiological responses i.e. multiunit activity of mitral cells was studied in rats which were submitted to a bilateral transection of LOT. The characteristic emotional reactions elicited in sham-operated rats by biologically meaningful odorants--odors of predator or of conspecific--were no longer observed after such lesion. At the olfactory bulb level, the differential habituation of mitral cell electrical responses according to the biological meaning of the stimuli disappeared in the rats with a bilateral section of LOT. However the awaking influence of the odorants when stimulations occurred during slow wave sleep remained unchanged: all the rats were more often awakened by the odor of predator than by the other stimuli. On the other hand, an inhibitory centrifugal influence of LOT on mitral cell electrical responses was noted in rats with a unilateral transection of LOT. The results are discussed in relation to the two olfactory pathways (lateral and medial). During wakefulness, LOT seems essential to the olfactory information processing; in slow wave sleep, the medial olfactory pathways appear to mediate olfactory discrimination and to elicit adapted arousal. PMID- 6293520 TI - The role of the medial olfactory pathways in olfaction: behavioral and electrophysiological data. AB - The role of the olfactory inputs transmitted by the medial olfactory pathways - i.e. anterior limb of the anterior commissure and medial forebrain bundle-was studied in rats submitted either to a bilateral lesion of the anterior commissure (bAC group) or to a bilateral transection of the lateral olfactory tract followed by an anterior commissure bilateral lesion (bCL group). In the bCL group, the characteristic emotional reactions usually observed in sham-operated rats to biologically meaningful odorants (odor of a predator or of conspecifics) were no longer observed; in this group, the modulation of mitral cell electrical responses according to the biological meaning of the stimuli disappeared and no habituation was noted when the stimulations were repeated: few responses appeared at random; lastly, all the stimuli tested, the odor of conspecifics excepted, did not exert an awaking influence on the sleeping rats (slow wave sleep). On the contrary, emotional reactions were always observed in the bAC group, a slight increase of reactivity to the odorants was even noted. In this group of animals, the anterior commissure bilateral lesion induced a large increase of the bulbar responses to all the odorants; the habituation of the electrical responses disappeared, a large percentage of responses being always noted even for the tenth series of stimulation; these results point out the great inhibitory influence that the anterior commissure exerts on the olfactory bulb activity. In these bAC rats almost usual awaking influences were noted, however, the odor of conspecifics became as efficient as fox odor to awake the animals. Results are discussed in relation to the functional role of the medial and lateral olfactory pathways, the integrity of both the pathways being necessary for an accurate olfactory process. PMID- 6293521 TI - Pre- and postjunctional blocking effects of aminoglycoside, polymyxin, tetracycline and lincosamide antibiotics. AB - The effects of seven antibiotics (streptomycin, amikacin, polymyxin B, lincomycin, clindamycin, tetracycline and oxytetracycline) were compared with those of magnesium, tubocurarine and lignocaine in the frog sciatic nerve- sartorius muscle preparation, using intracellular recording techniques. All compounds except tubocurarine decreased end-plate potential quantal content. The prejunctional effects of magnesium, streptomycin, amikacin, polymyxin B and oxytetracycline (but not the other drugs) were well reversed by increasing the calcium concentration. At concentrations which depressed quantal content, only magnesium, tetracycline and oxytetracycline did not reduce postjunctional sensitivity. Further postjunctional effects of the drugs were revealed by alterations in the time-courses of end-plate potentials. All the drugs tested except magnesium, tubocurarine and lincomycin produced changes in muscle action potentials. None of the compounds had anticholinesterase activity. The results confirm that aminoglycoside, polymyxin, tetracycline and lincosamide antibiotics produce neuromuscular block by a combination of both pre- and postjunctional actions. PMID- 6293522 TI - In vitro comparison between the neuromuscular and ganglion blocking potency ratios of atracurium and tubocurarine. AB - Results from in vitro experiments, using the hypogastric nerve--vas deferens preparation, and the phrenic nerve--hemidiaphragm preparation of the guineapig, have been used to determine the separation between the neuromuscular and ganglionic blocking effects of atracurium and tubocurarine. Regression lines were used to calculate the concentrations of each drug (99% confidence limits) which would produce a 50% blockade (EC50) of ganglionic and neuromuscular transmission. The equipotent molar ratio, using EC50 values, for ganglionic/neuromuscular blockade was 48 for atracurium and 9.4 for tubocurarine. PMID- 6293524 TI - Antibodies from EB-virus-transformed lymphocytes of lymph nodes adjoining lung cancer. PMID- 6293525 TI - Modifications of the hydroxylapatite assay to measure oestrogen receptors in human breast carcinomas. PMID- 6293523 TI - Epstein-Barr virus in nasopharyngeal and salivary gland carcinomas of Greenland Eskimoes. AB - Biopsy specimens from nasopharyngeal carcinomas (NPC) or salivary-gland carcinomas (SGC) in Greenland Eskimoes were examined for the presence of Epstein Barr virus (EBV) DNA and sera from the patients were tested for EBV-specific antibody titres. Six out of 7 NPCs and one from an undifferentiated SGG were positive for EBV DNA. The EBV-specific antibody spectra and titres of the patients with NPC or undifferentiated SGG conformed to the results of earlier studies in other high-incidence areas. PMID- 6293526 TI - Antibody to Herpesvirus hominis in patients with carcinoma of the cervix. PMID- 6293527 TI - Types I and III collagens and the activities of prolyl hydroxylase and galactosylhydroxylysyl glucosyltransferase in skin lesions of tuberous sclerosis. AB - Collagen synthesis and the ratios of collagen types I and III were assayed from the skin lesions of five subjects with tuberous sclerosis. Collagen synthesis, measured by the activities of prolyl hydroxylase and galactosylhydroxylysyl glucosyltransferase, was clearly increased in three angiofibromas of these patients and in one soft tumour of the face, but it was unchanged in shagreen patches. The total collagen content was decreased in angiofibromas, indicating either increased turnover of collagen or an increased amount of cellular or other macromolecular elements in these lesions. The proportions of types I and III collagens, estimated by cyanogenbromide digestion and SDS-gel electrophoresis, were 80-90% and 10-20%, respectively, in all samples except two angiofibromas, in which the relative amount of type III collagen was increased. This may indicate that angiofibromas of tuberous sclerosis are heterogenous with respect to the collagen types they contain, and that there may be disturbed cell growth or collagen synthesis, with individual variation from case to case. PMID- 6293528 TI - Malignant eccrine poroma: a study of twenty-seven cases. PMID- 6293529 TI - Pityrosporum folliculitis and ketoconazole. PMID- 6293530 TI - Rh immunization facilitated by passively-administered anti-Rh? AB - The response to 0.8 ml of D-positive (ccDEE) red cells injected i.v., together with 1 microgram anti-D i.m., was studied in 13 D-negative male volunteers (test subjects); the red cells were labelled with 51Cr and were found to have a mean survival time of 24 d, after correction for Cr elution. Within the following 5 months, nine of the 13 subjects made anti-D, compared with four out of 12 control subjects in a previous series who received 1 ml of red cells from the same donor but without passively-administered antibody. The time at which anti-D was first detected and the maximum concentration of anti-D attained were similar in test and control subjects. After a second injection of D-positive red cells, given 5 months after the first, the total number of subjects making anti-D became 11 out of 13 in the test series, compared with six out of 12 in the previous control series. Antibody concentrations were similar in test and control subjects. Evidently, Rh immunization was not augmented if this term implies accelerated formation of antibody or an obvious increase in antibody concentration. On the other hand, the suggestive increase in the proportion of responders, though not statistically significant, is consistent with the earlier observations of others and suggests that the effect of a small dose of passively administered antibody is to facilitate primary immunization. PMID- 6293531 TI - Clinical and aetiological study of adenoviral conjunctivitis, with special reference to adenovirus types 4 and 19 infections. AB - The clinical and aetiological findings are presented on 343 patients with clinical adenoviral conjunctivitis treated between July 1979 and July 1980 at 3 eye clinics in Sapporo. The age of the patients ranged from 12 days to 79 years, and the monthly incidence of the disease from 9 to 83, with clustering in the summer season. The aetiological diagnosis was established in 196 (57%) of 343 patients: adenovirus 3 in 24 cases; adenovirus 4 in 33 cases, including one case serologically diagnosed; adenovirus 8 in 124 cases; and adenovirus 19 in 15 cases. The different serotypes caused different clinical pictures. Adenovirus 19 conjunctivitis was more severe, with keratitis and preauricular lymphadenopathy more frequent than that of adenovirus 3 conjunctivitis. Adenovirus 4 conjunctivitis was generally similar to adenovirus 3 conjunctivitis. The clinical difference between the 2 groups adenoviruses 3 and 4, and adenoviruses 8 and 9, was statistically significant. Although in Japan adenovirus 8 was the most prevalent, adenoviruses 4 and 19 should be considered as causative agents of adenoviral conjunctivitis. PMID- 6293533 TI - Clinical and epidemiological features of adenovirus keratoconjunctivitis in London. AB - Ninety-eight patients were studied. Ninety were consecutive patients who were isolation-positive for adenovirus, and 8, who were associated with a hospital outbreak of adenovirus serotype 8 infection, developed characteristic features of infection but were isolation-negative. The ratio of males to females was 2 to 1, and most patients were aged 20 to 39. Adenovirus serotypes 3, 7, and 8 were isolated from 86% of patients, and serotypes 2, 4, 5, 11, 15, and 15/29 from the remaining 14%. Adenovirus serotype 7 was more commonly isolated from patients under the age of 19 and was Not isolated during winter. Sources of infection could be identified in 36% of patients and included contact with upper respiratory tract of ocular infections, a hospital outbreak, and a recent visit to a swimming pool. Associated systemic disease was detected in 47% of patients, most of whom had upper respiratory tract infection. The most severe and prolonged conjunctivitis was caused by serotypes 5 and 8. Most patients developed epithelial punctate keratitis. Subepithelial punctate keratitis, which was once considered to be a characteristic feature of adenovirus serotype 8, developed in cases of serotype 3, 4, 5, 7, and 8 infection. PMID- 6293532 TI - Beta adrenergic receptors in pigmented ciliary processes. AB - Beta adrenergic receptors from membrane fragments of pigmented sheep eyes were studied and characterised by ligand binding techniques after the removal of melanin. In a representative experiment the beta max (total number of beta receptors) was 394.9 fmol/mg protein. The receptor affinity (Ka) was 440 pM. The potency series of drugs to displace 125I-HYP from the receptors was timolol = ( ) propranolol greater than (+) propranolol greater than salbutamol greater than practolol. beta 1 Receptors were not detected in the ciliary processes. beta 2 Receptors were the prominent adrenergic receptors present. The theory as to how beta blockers work in glaucoma, their site of action, and the potential role of beta 2 blockers for use in intraocular pressure control is discussed. PMID- 6293534 TI - Case report of granular cell myoblastoma arising within the medial rectus muscle. PMID- 6293535 TI - The effect of heat treatment and particle size of bran on mineral absorption in rats. AB - The effect of heat treatment of bran on the true zinc absorption was measured using an isotope-dilution technique. A bran-based breakfast cereal (heated to 204 degrees for 40 min during manufacture) was incorporated into a semi-synthetic diet at a level of 180 g/kg. A parallel diet was formulated containing an identical weight of untreated bran from the same source plus other ingredients used to make the cereal. 2. Young male Wistar rats (mean weight 80 g) were injected intramuscularly with 65Zn to label body Zn. They were given the heat treated- and untreated-bran diets for 9 d. During the last 6 d of this period Zn intakes and faecal and urinary Zn were measured in order to calculate apparent Zn retention. True Zn retention was measured by taking into account losses of Zn of endogenous origin (labelled with 65Zn), by measuring faecal and urinary radioactivity of endogenous origin. 3. Heat treatment of bran removed approximately one-third of the phytate, but this was not enough to improve Zn absorption from the diet. True Zn retention measured by isotope dilution was significantly higher (P less than 0.02) than apparent Zn retention measured by the conventional balance technique. 4. The hypothesis that a reduction in particle size of bran would improve mineral availability was tested by feeding coarse and milled bran (100 mg/kg diet) in a semi-synthetic diet to rats and measuring true Fe and apparent Zn absorptions. The importance of phytate was also investigated by feeding a diet containing dephytinized bran. 5. Male Wistar rats (mean weight 172 g) were given diets containing coarse, milled or dephytinized bran for 9 d. Fe and Zn intakes were measured and faeces and urine collected for Fe and Zn analysis. 6. Rhe mean (+/- SE) particle size of the bran was reduced on milling from 3.5 (+/- 1.8) to 0.2-0.5 mm. There were no differences in the fraction of Fe retained between the three groups. Particle size had a small effect on Zn retention which was marginally higher in rats on the milled-bran diet (0.126 (+/- 0.023)) than in those on the coarse-bran diet (0.087 (+/- 0.012)). Total removal of phytate had a greater effect and apparent Zn retention from the dephytinized-bran diet was significantly higher (0.182 (+/- 0.027), P less than 0.001). PMID- 6293536 TI - Correlations between structural and spectroscopic properties of the high-spin heme protein cytochrome c'. AB - The cytochromes c' are a class of heme proteins whose native spectroscopic properties have been suggested to represent a quantum mechanical admixture of intermediate-(S = 3/2) and high-(S = 5/2) spin states. Here features of the cytochrome c' heme environment, as revealed by X-ray crystallographic studies of the dimeric cytochrome c' from Rhodospirillum molischianum, are related to the observed spectroscopic properties. The environment of the heme group in cytochrome c' supports the existence of the admixed spin state at neutral pH and suggests that pH-dependent transition to a pure high-spin state at alkaline pH involves deprotonation of the histidine axial ligand to the heme iron. PMID- 6293537 TI - Adenosine 3',5'-cyclic monophosphate protein kinase from bovine brain: inactivation of the catalytic subunit and holoenzyme by 7-chloro-4-nitro-2,1,3 benzoxadiazole. PMID- 6293538 TI - Activation and inactivation kinetics of Torpedo californica acetylcholine receptor in reconstituted membranes. AB - By use of a quench-flow technique to measure tracer ion flux rates in a physiologically significant time domain, the kinetics of activation and inactivation of purified reconstituted acetylcholine receptor (AChR) were investigated. After solubilization in sodium cholate, purification by affinity chromatography, and reconstitution into soybean lipids, the AChR from Torpedo californica displayed a characteristically fast rate of ion influx measured with 86Rb+. At 4 degrees C 1 mM carbamoylcholine (Carb) stimulated a fast (t1/2 = 7 ms) first-order filling of vesicle internal volume that presented a 10(4)-fold stimulation of ion flux rate by Carb. The concentration dependence of activation was sigmoidal with a half-maximal value at 3 X 10(-4) M Carb. In the presence of Carb, the purified AChR also underwent a two-step inactivation (desensitization) process. Inactivation was measured by preincubating AChR with Carb for various times (milliseconds to minutes) and then measuring the 86Rb+ influx rate. The two inactivation processes were each characterized by a distinct maximum rate (5.3 and 0.10 s-1) and by a different dependence on Carb concentration. The slow phase of inactivation gave a half-maximal rate at 2.5 X 10(-4) M Carb, and the fast inactivation was half-maximal at 1.3 X 10(-3) M Carb. The concentration dependence curves for both inactivation processes were approximately hyperbolic. The results are discussed in terms of models that describe the relationship between ligand binding and the processes of channel activation and desensitization. PMID- 6293539 TI - Ribulose-1,5-bisphosphate carboxylase: enzyme-catalyzed appearance of solvent tritium at carbon 3 of ribulose 1,5-bisphosphate reisolated after partial reaction. AB - When ribulose 1,5-bisphosphate is allowed to react with carbon dioxide in tritiated water in the carboxylation reaction catalyzed by ribulose-1,5 bisphosphate carboxylase from Rhodospirillum rubrum, the ribulose 1,5 bisphosphate reisolated after partial reaction is found to be labeled. The specific radioactivity of the remaining substrate pool rises during the course of the reaction. Experiments in deuterium oxide show that the isotopic label resides on carbon 3. Earlier failures to detect this exchange process probably derive from the use of enzyme that was, in the absence of carbon dioxide, inactive. The present results provide direct evidence for the intermediacy of the enediol between C-2 and C-3 of ribulose 1,5-bisphosphate and show that the enolization step is at least partially rate limiting in the overall carboxylase reaction. The specific radioactivity of the product 3-phospho-D-glycerate remains constant throughout the course of the reaction at about one-sixth that of the solvent. This strengthens the argument against the involvement of "sticky" protons in the reaction. PMID- 6293540 TI - Conformations of cytochrome oxidase: thermodynamic evaluation of the interconversion of the 418- and 428-nm forms. AB - Oxidized cytochrome c oxidase exists in two reasonably well-defined conformations, a high-spin conformation with maximal absorption at 418 nm and a low-spin conformation with maximal absorption at 428 nm. The equilibrium between these two conformations has been studied as a function of pH, pressure, and temperature. pH effects the equilibrium between the two conformations, the maximum fraction of the 418-nm form being found at about pH 6.8. Increasing pressure displaced the equilibrium toward the 428-nm form; the molar volume changes found are independent of pH but strongly dependent on temperature. Increasing temperature over the range -20 to 25 degrees C displaces the equilibrium toward the 428-nm form; the van't Hoff plots that result show a discontinuity at about 10 degrees C. Above 10 degrees C, delta H is relatively constant as a function of pH; below 10 degrees C, delta H is strongly pH dependent. Delta G, delta H, delta S, and delta V have been evaluated for the equilibrium. PMID- 6293541 TI - Purification and characterization of human lymphoid poly(adenosine diphosphate ribose) polymerase. AB - Poly(ADP-ribose) polymerase has been purified 12 000-fold from human tonsils with an 83% recovery of enzymatic activity relative to that of the initial homogenate. The specific activity of the purified enzyme is 862 units/mg of protein. The isolated protein has a molecular weight of approximately 116 000 as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The apparent Km for NAD+ is estimated to be 185 microM at pH 8.0 and 37 degrees C. The purified enzyme has an absolute requirement for exogenous DNA for catalytic activity, and the reaction is enhanced by the addition of purified histone H1. The enzyme does not require magnesium or other divalent cations for activity. Enzyme activity is inhibited by p-(hydroxymercuri)benzoate and N-ethylmaleimide. Thymidine, theophylline, nicotinamide, and 5-methylnicotinamide markedly inhibit enzyme activity whereas ADP-ribose, 3',5'-cAMP, and sodium fluoride have a minimal effect on enzyme activity. Autoradiograms of labeled products of the reaction catalyzed by the purified enzyme at different concentrations of NAD+ and at different incubation times show that at low concentrations of NAD+ and after short incubations, poly(ADP-ribosyl)ation of the enzyme occurs preferentially over that of histone H1; at higher concentrations of NAD+ or after longer incubations, poly(ADP-ribosyl)ation of histone H1 is increased. PMID- 6293542 TI - Hydroxylation of prolyl residues in type II procollagen in vitro and in cellulo. Lack of preferential hydroxylation of specific regions of the protein. AB - [14C]Proline-labeled protocollagen, the unhydroxylated form of procollagen, was isolated from cartilage cells incubated with alpha, alpha'-dipyridyl. For examination of the initial steps in the hydroxylation of the protein, it was incubated in vitro with prolyl hydroxylase so that an average of 1.3-2.7 prolyl residues per chain was hydroxylated. The partially hydroxylated alpha chain were cleaved with cyanogen bromide, and the fragments were separated by polyacrylamide gel electrophoresis or column chromatography. The cyanogen bromide fragments were hydroxylated to the same degree. The results indicated, therefore, that in the initial hydroxylation of alpha chains in vitro, there was no preferential hydroxylation of any specific regions of the protein. In a second series of experiments, cartilage cells were incubated with [14C]proline and alpha, alpha' dipyridyl so that prolyl hydroxylase in the cells was extensively, but not completely, inhibited. Partially hydroxylated alpha chains were isolated, and cyanogen bromide fragments of the alpha chains from the cells were assayed for hydroxy[14C]proline. The alpha chains contained an average of two residues of hydroxyproline per chain, and the cyanogen bromide fragments were hydroxylated to about the same degree. The results indicated, therefore, that when prolyl hydroxylase activity in cells is low relative to the rate at which pro alpha chains are synthesized, hydroxylation of prolyl residues occurs as it does in vitro, and there is no preferential hydroxylation of a specific region of the protein. PMID- 6293543 TI - Metal ion binding to alpha-lactalbumin species. AB - A strong cation (calcium) binding site has been demonstrated to exist in several alpha-lactalbumin species; bovine, goat, human, and guinea pig. A metal ion induced conformational change occurs, resulting in a unique (10-14-nm) blue shift and relative quenching of Trp fluorescence for all species. Calcium ion binding to the alpha-lactalbumins yielded dissociation constants (Kdiss consistently in the 10(-10)--10(-12) M range, while Mn(II) binding was in the 20-30 microM range. Independent determinations of these cation binding equilibria were made by ESR measurements of free unliganded Mn(II) in titrations with the bovine species. One strong site (Kdiss = 30.5 microM) was found, which correlated directly with the fluorescence-associated cation binding, plus three weaker sites (Kdiss = 1.1, 5.0, and 5.0 mM, respectively). Several lanthanides as well as Mg(II) were found to displace Mn(II) from the strong site on bovine alpha-lactalbumin (as monitored by ESR) and to cause the identical fluorescence changes as found for Ca(II) and Mn(II) above. The importance of measuring these equilibria by both fluorescence and ESR was borne out by demonstrating the potential errors in estimating dissociation equilibria by the fluorescence method alone. Also, the errors in estimating Kdiss for samples containing partially metal bound apo-alpha lactalbumin are described as well as rapid, sensitive methods for estimating the extent of metal-free protein and correctly accounting for residual bound metal in equilibrium calculations. PMID- 6293544 TI - Subunit structure of islet-activating protein, pertussis toxin, in conformity with the A-B model. AB - The subunit structure of islet-activating protein (IAP), pertussis toxin, has been analyzed to study a possibility that this protein is one of the A-B toxins [Gill, D. M. (1978) in Bacterial Toxins and Cell Membranes (Jeljaszewicz, J., & Wadstrom, T., Eds.) pp 291-332, Academic Press, New York]. Heating IAP with 1% sodium dodecyl sulfate caused its dissociation into five dissimilar subunits named S-1 (with a molecular weight of 28 000), S-2 (23 000), S-3 (22 000), S-4 (11 700), and S-5 (9300), as revealed by polyacrylamide gel electrophoresis; their molar ratio in the native IAP was 1:1:1:2:1. The molecular weight of IAP estimated by equilibrium ultracentrifugation was 117 000 which was not at variance with the value obtained by summing up molecular weights of the constituent subunits. The preparative separation of these IAP subunits was next undertaken; exposure of IAP to 5 M ice-cold urea for 4 days followed by column chromatography with carboxymethyl-Sepharose caused sharp separation of S-1 and S 5, leaving the other subunits as two dimers. These dimers were then dissociated into their constituent subunits, i.e., S-2 and S-4 for one dimer and S-3 and S-4 for the other, after 16-h exposure to 8 M urea; these subunits were obtained individually upon further chromatography on a diethylaminoethyl-Sepharose column. Subunits other than S-1 were adsorbed as a pentamer by a column using haptoglobin as an affinity adsorbent. The same pentamer was obtained by adding S-5 to the mixture of two dimers. Neither this pentamer nor other oligomers (or protomers) exhibited biological activity in vivo. Recombination of S-1 with the pentamer at the 1:1 molar ratio yielded a hexamer which was identical with the native IAP in electrophoretic mobility and biological activity to enhance glucose-induced insulin secretion when injected into rats. In the broken-cell preparation, S-1 was biologically as effective as the native IAP; both catalyzed ADP-ribosylation of a protein in membrane preparations from rat C6 glioma cells. In conclusion, IAP is an oligomeric protein consisting of an A (active) protomer (the biggest subunit) and a B (binding) oligomer which is produced by connecting two dimers by the smallest subunit in a noncovalent manner. Rationale for this terminology is discussed based on the A-B model. PMID- 6293545 TI - Stoichiometry of the H+-ATPase of growing and resting, aerobic Escherichia coli. AB - The H+/ATP stoichiometry of the proton-translocating ATPase was investigated in growing and nongrowing, respiring cells of Escherichia coli. The protonmotive force, delta p, was determined by measuring the transmembrane chemical gradient of protons, delta pH, from the cellular accumulation of benzoate anions, and the electrical gradient, delta psi, from the accumulation of the lipophilic cation tetraphenylphosphonium (TPP+). The accumulation of lactose was also used to calculate the delta p in this lactose operon constitutive beta-galactosidase negative mutant. The phosphorylation potential, delta GP', was determined by measuring the cellular concentration of ATP, ADP, and inorganic phosphate. According to chemiosmotic principles, at steady state the phosphorylation potential is in thermodynamic equilibrium with the protonmotive force, and thus the ratio delta p/delta GP' can be used to determine the H+/ATP ratio. Respiring E. coli cells, in mid-exponential phase of growth or incubated in buffer, at external pHs from 6.25 to 8.25 had a constant delta GP' of about 500 mV. The H+/ATP ratio was found to be 3 when the delta p value derived from lactose accumulation levels was used. However, when the delta p values derived from delta pH and delta psi were used in the calculations, the H+/ATP ratio varied from about 2.5 at external pH 6.25 to about 4 at pH 8.25. Arguments are presented for the hypothesis that the delta psi values obtained from the TPP+ measurements are likely to be inaccurate and that a value of 3 H+/ATP, independent of the external pH, is likely to be the valid stoichiometry. PMID- 6293546 TI - Reconstitution of types I and II adenosine cyclic 3',5'-phosphate dependent protein kinase. AB - Fluorescence intensity and anisotropy measurements using the fluorescent adenosine cyclic 3',5'-phosphate (cAMP) analogue 1,N6-ethenoadenosine cyclic 3',5'-phosphate (epsilon-cAMP) are sensitive to the dissociation of epsilon-cAMP which occurs when either the type I or the type II regulatory subunit (RI or RII) of cAMP-dependent protein kinase associates with the catalytic subunit. Studies using epsilon-cAMP show that MgATP has opposite effects on the reconstitution of both types of protein kinase: MgATP strongly stabilizes the type I holoenzyme while it slightly destabilizes the type II holoenzyme. The synthetic substrate Kemptide has a small inhibitory effect on the reconstitution of both holoenzymes when tested at 10 microM concentration. The protein kinase inhibitor has a larger effect which is especially pronounced in the reassociation of the type I enzyme. The diminished relative ability of the type I regulatory subunit to compete with the protein kinase inhibitor suggests that the combined effects of the two opposing equilibria (epsilon-cAMP and catalytic subunit binding) are different for the two types of regulatory subunits. Displacement experiments show that cAMP and epsilon-cAMP bind about equally well to the type I subunit. Slow conformational changes accompanying the binding of epsilon-cAMP by both regulatory subunits are greatly accelerated with the holoenzymes, suggesting that dissociation of the holoenzymes occurs via ternary complexes. The time courses of epsilon-cAMP binding also show the heterogeneity of binding characteristics of RII. The 37 000-dalton fragment of type II subunit retains the epsilon-cAMP binding properties of the native subunit. However, only a fraction of the fragment preparation (approximately 32% estimated from sedimentation measurements) binds the catalytic subunit well, suggesting heterogeneity of cleavage. PMID- 6293547 TI - Binding of 5,5'-bis[8-(phenylamino)-1-naphthalenesulfonate] by the regulatory subunits of adenosine cyclic 3',5'-phosphate dependent protein kinase. AB - Binding to the regulatory subunits of types I and II adenosine cyclic 3',5' phosphate (cAMP) dependent protein kinase (RI and RII, respectively) produces large distinctive increases in fluorescence and optical activity of 5,5'-bis[8 (phenylamino)-1-naphthalenesulfonate] [bis(ANS)]. Both specific and nonspecific interactions are involved. Association of the regulatory subunits with either the catalytic subunit or cAMP results in dissociation of a major portion of the bound bis(ANS) as detected by changes in fluorescence and circular dichroism. The results are consistent with the accepted cAMP binding properties of RI and RII, showing cooperativity in case of RI and two heterologous binding sites for RII. cGMP has the same overall effect on bis(ANS) binding as cAMP. However, very high concentrations are required for complete dissociation of bis(ANS) from RII, consistent with the observation that cGMP is inefficient in bringing about the dissociation of the type II holoenzyme. Magnesium binding to sites having dissociation constants of ca. 12 mM increases the interaction of bis(ANS) with both of the isolated regulatory subunits. Experiments involving the 37 000-dalton fragment of RII indicate that the limited proteolytic cleavage was heterogeneous, with only 24-39% of the resulting population interacting strongly with the catalytic subunit. PMID- 6293548 TI - Partial purification and characterization of phosphotyrosyl-protein phosphatase from Ehrlich ascites tumor cells. AB - We have previously described a phosphotyrosylprotein phosphatase in membrane vesicles from human epidermoid carcinoma A431 cells which is inhibited by micromolar concentration of Zn2+ and is insensitive to ethylenediaminetetraacetic acid (EDTA) and NaF [Brautigan, D. L., Bornstein, P., & Gallis, B. (1981) J. Biol. Chem. 256, 6519-6522]. Here we present the identification and partial purification of a similar enzyme from lysates of Ehrlich ascites tumor cells. the enzyme was purified by using diethylaminoethyl-Sephadex, Zn2+ affinity, and Sephadex G-75 chromatography. During purification, the phosphatase was separated into at least three fractions, all of which exhibited very similar properties and an apparent molecular weight of 40 000 upon gel filtration. The enzyme dephosphorylated phosphotyrosine (P-Tyr)-containing carboxymethylated and succinylated (CM-SC) phosphorylase with an apparent Km of 0.8 microM, as well as P-Tyr containing casein and epidermal growth factor (EGF) receptor kinase, but did not dephosphorylate P-Ser-phosphorylase. The phosphatase was inhibited by Zn2+ at micromolar concentrations (K0.5 with EGF receptor kinase = 5 X 10(-6) M; with CM-SC phosphorylase = 3.3 X 10(-5) M) but not by millimolar concentrations of EDTA and NaF. No inhibition was seen with 1 mM tetramisole, a specific inhibitor of alkaline phosphatases. P-Tyr inhibited the enzyme by 50% at 0.4 X 10(-3) M, while Tyr, Pi, PPi, and p-nitrophenyl phosphate, an excellent substrate for alkaline phosphatases and structurally very similar to P-Tyr, exerted partial inhibition at concentrations above 10(-3) M. The pH optimum was found to be 6.5 7, depending on the substrate used. Very little activity was seen below pH 5 and above pH 8.5. These properties clearly distinguish this enzyme from alkaline phosphatases, as well as the neutral and acidic protein phosphatases so far described, and therefore define it as a new enzyme of the phosphatase family--a phosphotyrosyl-protein phosphatase. PMID- 6293549 TI - Purification and properties of pyruvate dehydrogenase phosphatase from bovine heart and kidney. AB - Pyruvate dehydrogenase phosphatase was purified to apparent homogeneity from bovine heart and kidney mitochondria. The phosphatase has a sedimentation coefficient (S20,w) of about 7.4 S and a molecular weight (Mr) of about 150 000 as determined by sedimentation equilibrium and by gel-permeation chromatography. The phosphatase consists of two subunits with molecular weights of about 97 000 and 50 000 as estimated by sodium dodecyl sulfate--polyacrylamide gel electrophoresis. Phosphatase activity resides in the Mr 50 000 subunit, which is sensitive to proteolysis. The phosphatase contains approximately 1 mol of flavin adenine dinucleotide (FAD) per mol of protein of Mr 150 000. FAD is apparently associated with the Mr 97 000 subunit. The function of this subunit remains to be established. The phosphatase binds 1 mol of Ca2+ per mol of enzyme of Mr 150 000 at pH 7.0, with a dissociation constant (Kd) of about 35 microM as determined by flow dialysis. Use of ethylene glycol bis(beta-aminoethyl ether)-N,N,N',N' tetraacetate (EGTA) at pH 7.6 in conjunction with flow dialysis gave a Kd value for Ca2+ of about 8 microM. In the presence of both the phosphatase and the dihydrolipoyl transacetylase (E2) core of the pyruvate dehydrogenase complex, two equivalent and apparently non-interacting CA2+-binding sites were detected per unit of Mr 150 000, with a Kd value of about 24 microM in the absence and about 5 microM in the presence of EGTA. In the presence of 0.2 M KCl, which inhibits phosphatase activity about 95%, the phosphatase exhibited only one Ca2+-binding site, even in the presence of E2. The phosphatase apparently possesses an "intrinsic" Ca2+-binding site, and a second Ca2+-binding site is produced in the presence of E2. The second site is apparently altered by increasing the ionic strength. It is proposed that the second site may be at the interface between the phosphatase and E2, with Ca2+ acting as a bridging ligand for specific attachment of the phosphatase to E2. PMID- 6293550 TI - Orientation and motion of spin-labels in rabbit small intestinal brush border vesicle membranes. AB - The temperature dependence of the packing (order) and fluidity (microviscosity) of rabbit small, intestinal brush border vesicle membranes and of liposomes made from their extracted lipids has been investigated by using a variety of lipid spin probes. The lipids in the brush border membrane are present essentially as a bilayer. Compared to other mammalian membranes, the brush border membrane appears to be characterized by a relatively high packing order as well as microviscosity. At body temperature, the lipid molecules undergo rapid, anisotropic motion, which is essentially a fast rotation about an axis approximately perpendicular to the bilayer normal. Both the order (motional anisotropy) and the microviscosity increase with decreasing temperature and with increasing distance from the center of the bilayer. Qualitatively similar motional or fluidity gradients have been reported for other mammalian and bacterial membranes. The liposomes made from the extracted lipids have a somewhat lower packing order and a slightly higher fluidity than brush border vesicle membranes. The differences are, however, small indicating that the packing and the fluidity (microviscosity) of the membrane are primarily determined by the lipid composition. Membrane-associated proteins and cytoskeleton cannot play a dominant role in determining the order and fluidity of the lipid bilayer. Discontinuities are observed in the temperature dependence of various spectral parameters, the order parameter S, the rotational correlation time tau, and 2,2,6,6-tetramethylpiperidinyloxy partitioning. They are assigned to phase transitions and/or phase separations of the membrane lipids. These discontinuities occur at about 30, 20, and 13 degrees C for 5-doxyl-, 12-doxyl-, and 16-doxylstearic acid, respectively. The apparent transition temperature depends on the location of the spin probe along the bilayer normal, being higher the closer the probe is to the membrane surface. This indicates the possibility that chain melting is progressive and spreads with increasing temperature from the center of the membrane outward. PMID- 6293551 TI - Specific binding of toxin II from Centruroides suffusus suffusus to the sodium channel in electroplaque membranes. AB - The binding of toxin II from the scorpion Centruroides suffusus suffusus (CssII) to electroplaque membranes from Electrophorus electricus was studied with the use of a radiolabeled derivative of the toxin ([125I]CssII). Specific binding of the latter to the membranes required the protonation of a group, either in the membrane or in the toxin itself, with an apparent pKa value of 7.5 and also the presence of a certain minimum concentration of ions, though there was no requirement for a specific ion. At 20 degrees C and pH 6 the second-order rate constant for formation of the [125I]CssII-membrane complex was about 5 X 10(6) M 1 s-1, while the first-order constant for its dissociation was about 2 X 10(-3) s 1. Under equilibrium conditions specific binding of [125I]CssII was a simple saturable function of [125I]CssII concentration, characterized by a dissociation constant of 0.4-0.7 nM and a maximum capacity of 0.9-2.4 pmol of toxin/mg of membrane protein. The latter value was the same as the number of membrane sites that could specifically bind a radiolabeled derivative of tetrodotoxin. Unlabeled CssII displaced bound [125I]CssII with an apparent dissociation constant of about 1 nM. None of 19 other neurotoxins or local anaesthetics known to interact with Na+ channels in excitable cells affected [125I]CssII binding, but it was completely inhibited by toxin gamma from the scorpion Tityus serrulatus serrulatus. These findings suggest that the Na+ channel possesses a distinct class of binding sites to which these two scorpion toxins bind with high affinities. On the other hand, no CssII receptor was detected in crab axonal membranes, indicating that it is not a characteristic feature of all Na+ channels. PMID- 6293552 TI - Lactose-proton symport by purified lac carrier protein. AB - The lac carrier protein of Escherichia coli was purified by an improved procedure and its activity assayed by a rapid filter method. Following reconstitution of the carrier by octyl glucoside dilution, proteoliposomes were concentrated by filtration on a microporous filter. Lactose accumulation by adsorbed or entrapped proteoliposomes is driven by an artificially imposed pH gradient (interior alkaline), by a membrane potential (interior negative), or by a combination of both forces. Activity is almost completely abolished by the protonophore carbonyl cyanide m-chlorophenylhydrazone or by the competitive inhibitor thiodigalactoside. Addition of lactose to proteoliposomes under appropriate conditions results in alkalinization of the external medium. This effect is not observed with liposomes devoid of lac carrier or in the presence of proton conducting agents. The results provide a strong indication that the lac gamma gene product is the only protein in the cytoplasmic membrane of Escherichia coli required for lactose-proton symport. PMID- 6293553 TI - Photoaffinity labeling of human thyroxine-binding prealbumin with thyroxine and N (ethyl-2-diazomalonyl)thyroxine. AB - To facilitate studies of thyroid hormone-binding proteins, we have synthesized and tested the photoaffinity analogues N-(ethyl-2-diazomalonyl)-3,5,3' triiodothyronine (EDM-T3) and N-(ethyl-2-diazomalonyl)thyroxine (EDM-T4). The binding affinities of L-EDM-T4 and D-EDM-T4 to human thyroxine-binding prealbumin were 4% and 13.2%, respectively, that of L-thyroxine (L-T4). For comparison the affinities of L-EDM-T3 and D-EDM-T3 to crude rate liver nuclear receptor preparation were 0.1% and 0.7%, respectively, that of L-triiodothyronine (L-T3). Photolysis of prealbumin-[125I]-L-EDM-T4 complexes at 254 nm resulted in covalent linkage of [125I]-L-EDM-T4 to prealbumin as judged by sodium dodecyl sulfate gel electrophoresis. Virtually no labeling was observed in the absence of photolysis. Photolabeling of prealbumin was specific for the high-affinity hormone binding site since it was (a) completely blocked during photolysis in the presence of excess 3,5,3',5'-tetraiodothyroacetic acid, (b) saturated at high [125I]-L-EDM-T4 concentrations, (c) prevented when the hormone binding site had been previously blocked by dansylation of prealbumin, and (d) blocked competitively by T3 and T4 with inhibition constants (K1) similar to the dissociation constants (Kd) for these ligands. Analysis of prealbumin photolabeling by direct attachment of [125I]-L-EDM-T4 or attachment of unlabeled L-EDM-T4 followed by titration of the remaining sites with [125I]-L-T4 indicated a photolabeling efficiency of 54-61% at 63-67% site occupancy. After destruction of the diazo group by preirradiation, L-EDM-T4 was found to label prealbumin after further irradiation; 19-26% photolabeling efficiency could be achieved by using preirradiated reagent at 87 91% site occupancy. This carbene-independent photoattachment was also specific for a high-affinity hormone binding site. The mechanism of the carbene independent process may involve attachment via radical formation following photoinduced loss of the thyronine ring iodine. Accordingly, specific covalent cross-linking of L-T4 to prealbumin was demonstrated; however, the photolabeling efficiency was much lower than that with preirradiated EDM-T4. Both EDM-T4 and T4 were employed to photolabel prealbumin, thyroxine binding globulin, and albumin in unfractionated human serum. PMID- 6293554 TI - Interaction of a fluorescent N-dansylaziridine derivative of troponin I with calmodulin in the absence and presence of calcium. AB - Rabbit skeletal muscle troponin I was covalently labeled with N-dansylaziridine, resulting in a fluorescent labeled protein. This derivative (DANZTnI) and native troponin I (TnI) inhibited calmodulin (CaM) stimulation of bovine heart Ca2+ sensitive cyclic nucleodite phosphodiesterase with identical inhibition constants. Association of DANZTnI with calmodulin was monitored directly by changes in flourescence intensity in the presence of Ca2+ and by changes in fluorescence anisotropy in the absence of Ca2+. Quantitation of the affinity of calmodulin for calmodulin-binding proteins in both the presence and absence of Ca2+ is necessary for prediction of the extent of interaction of both Ca2+ and calmodulin-binding proteins with calmodulin in vivo. The dissociation constants for the DANZTnI-calmodulin-l4Ca2+ and DANZTnI-calmodulin complexes were 20 nM and 70 micrometers, respectively. These dissociation constants define a free energy coupling of-4.84 kcal/mol of troponin I for binding of Ca2+ and troponin I to calmodulin. The Ca2+ dependence for troponin I-calmodulin complex formation predicted from these experimentally determined parameters was closely approximated by the Ca2+ dependence for complex formation between troponin I and fluorescent 5-[[[(iodoacetyl)amino]ethyl]-amino]-1-napthalenesulfonic acid derivatized calmodulin as determined by fluorescence anisotropy. Complex formation occurred over a relatively narrow range of Ca2+ concentration, indicative of positive heterotropic cooperativity for Ca2+ and troponin I binding to calmodulin. PMID- 6293555 TI - High-affinity uptake and degradation of apolipoprotein E free high-density lipoprotein and low-density lipoprotein in cultured porcine hepatocytes. AB - Isolated pig liver membranes contain a specific "lipoprotein binding site" that recognizes low-density lipoproteins (LDL) and apolipoprotein E (apoE) free high density lipoprotein (HDL) [Bachorik, P. S., Kwiterovich, P. O., & Cooke, J. (1978) Biochemistry 17, 5287-5299]. We report here that a similar site exists in cultured porcine hepatocytes and that it mediates the uptake and degradation of apoE-free HDL. The binding of 125I-labeled HDL and 125I-labeled LDL (125I-HDL and 125I-LDL, respectively) at 4 degrees C and the uptake and degradation of the lipoproteins at 37 degrees C were time dependent and saturable and were not inhibited by unrelated proteins. Chloroquine (6 x 10(-5)M) inhibited the degradation of 125I-HDL by 76% and of 125I-LDL by greater than 99%; leupeptin inhibited the degradation of both lipoproteins by about 25%. 125I-HDL binding (4 degrees C), uptake, and degradation (37 degrees C) were inhibited by LDL, methyl LDL, and methyl-HDL about as well as by unlabeled HDL but were unaltered in Pronase-treated cells or in cells that were cultured for 24 h in either lipoprotein-free medium containing HDL or LDL (200 micrograms/mL). In contrast, these conditions affected the uptake and degradation of 125I-LDL disproportionately. HDL and methyl-LDL inhibited 125I-LDL uptake by 50% or more but had little effect on degradation. 125I-LDL binding was reduced by 12% and degradation by 57% in Pronase-treated cells. Preincubation of the cells with LDL (200 micrograms/mL) reduced uptake by 35% and degradation by 68%. Similar preincubation with HDL (200 micrograms/mL) increased 125I-LDL degradation by 60% but did not affect 125I-LDL uptake. The findings indicated the presence in porcine hepatocytes of at least two distinct sites for lipoproteins. One site resembled the LDL receptor and mediated 125I-LDL degradation. A second, Pronase insensitive site recognized both HDL and LDL. This site mediated almost all of the degradation of 125I-HDL but little if any degradation of 125I-LDL. PMID- 6293556 TI - Monomer of sodium and potassium ion activated adenosinetriphosphatase displays complete enzymatic function. AB - The distribution of sodium and potassium ion activated adenosinetriphosphatase [(Na+ + K+)-ATPase] among the various oligomeric forms present in a given solution is assessed unambiguously by cross-linking with glutaraldehyde. Purified enzyme dissolved in a solution of a nonionic detergent, octaethylene glycol dodecyl ether, remains dispersed and unaggregated after removal of the bulk of the detergent. Increases in the aggregation of the enzyme, which have been previously observed upon the addition of substrates to such a solution, are found to be due to changes in ionic strength rather than a consequence of the initiation of turnover. Furthermore, conditions are described that produce solutions containing stable, enzymatically active mixtures of the smaller oligomers of the asymmetric unit, alpha beta. Cross-linking by glutaraldehyde while the enzyme is turning over demonstrates that at least one of these oligomers is responsible for the observed enzymatic activity. A determination of which oligomers are present in each fraction from a glycerol gradient demonstrates that the profiles of the enzymatic activity and the concentration of monomer coincide. In addition, the monomer can form the sodium-dependent, phosphorylated intermediate of the mechanism for the enzyme. Finally, a preparation of (Na+ + K+)-ATPase, dissolved in solutions of the same nonionic detergent, can be prepared in which the predominant species (greater than 85%) is the monomer. The enzyme in this solution exhibits high specific activity, and its apparent Michaelis constants for the cationic substrates are very similar to those of the purified, membrane-bound enzyme. It is concluded from these results that a monomer of the alpha beta asymmetric unit is fully capable of catalyzing (Na+ + K+)-ATPase activity, and hence active transport, in the native enzyme. A reassessment of proposed molecular mechanisms for active transport is made in light of these discoveries. PMID- 6293557 TI - Kinetics of ubiquinol-1-cytochrome c reductase in bovine heart mitochondria and submitochondrial particles. AB - A kinetic study on ubiquinol-cytochrome c reductase (EC 1.10.2.2) has been undertaken either in situ in KCN-inhibited mitochondria and submitochondrial particles, or in the isolated cytochrome b-c1 complex using ubiquinol-1 and exogenous cytochrome c as substrates. The steady-state two-substrate kinetics of the reductase appears to follow a general sequential mechanism, allowing calculation of a Km for ubiquinol-1 of 13.4 microM in mitochondria and of 24.6 microM in the isolated cytochrome b-c1 complex. At low concentrations of cytochrome c, however, the titrations as a function of quinol concentration appear biphasic both in mitochondria and in submitochondrial particles containing trapped cytochrome c inside the vesicle space, fitting two apparent Km values for ubiquinol-1. Relatively high antimycin-sensitive rates of ubiquinol-1-cytochrome c reductase have been found in submitochondrial particles: both the Vmax and the Km for ubiquinol-1 are, however, affected by the overall orientation of the particle preparation, i.e., by the reactivity of cytochrome c with its proper site. The turnover numbers corrected for particle orientation with respect to cytochrome c interaction are at least 2-fold higher in submitochondrial particles than in mitochondria. This is particularly evident using inside-out particles containing trapped cytochrome c in the vesicle space (and therefore reacting with its physiological site). A diffusion step for the quinol substrate appears to be rate limiting in mitochondria and can be removed by addition of deoxycholate, suggesting that the oxidation site of ubiquinol may be more exposed to the matrix side of the inner mitochondrial membrane. PMID- 6293558 TI - Electron transfer after flash photolysis of mixed-valence carboxycytochrome c oxidase. AB - The light-induced difference spectra of the fully reduced (a2+ a23+-CO) complex and the mixed-valence carboxycytochrome c oxidase (a3+ a23+-CO) during steady state illumination and after flash photolysis showed marked differences. The differences appear to be due to electron transfer between the redox centres in the enzyme. The product of the absorbance coefficient and the quantum yield was found to be equal in both enzyme species, both when determined from the rates of photolysis and from the values of the dissociation constants of the cytochrome a23+-CO complex. This would confirm that the spectral properties of cytochrome a3 are not affected by the redox state of cytochrome a and CuA. When the absorbance changes after photolysis of cytochrome a23+-CO with a laser flash were followed on a time scale from 1 mus to 1 s in the fully reduced carboxycytochrome c oxidase, only the CO recombination reaction was observed. However, in the mixed valence enzyme an additional fast absorbance change (k = 7 X 10(3) s-1) was detected. The kinetic difference spectrum of this fast change showed a peak at 415 nm and a trough at 445 nm, corresponding to oxidation of cytochrome a3. Concomitantly, a decrease of the 830 nm band was observed due to reduction of CuA. This demonstrates that in the partially reduced enzyme a pathway is present between CuA and the cytochrome a3-CuB pair, via which electrons are transferred rapidly. PMID- 6293559 TI - Basal-lateral membranes from rabbit renal cortex prepared on a large scale in a zonal rotor. AB - Basal-lateral membranes from the renal cortex of the rabbit were isolated by sucrose gradient centrifugation in a zonal rotor which allows for a large-scale preparation of these membranes. A heterogeneous population of membranes (P4) which contained 29% of the (Na+ + K+)-ATPase found in the homogenate of renal cortex was prepared by differential centrifugation. When pellet P4 was subjected to centrifugation in a sucrose gradient the activity of (Na+ + K+)-ATPase, a marker for basal-lateral membranes, could be separated from enzymatic markers of other organelles. The specific activity of (Na+ + K+)-ATPase was enriched 12-fold at a density of 1.141 g/cm3. Membranes (P alpha) contained in the (Na+ + K+) ATPase-rich fractions consisted primarily of closed vesicles which exhibited probenecid inhibitable transport of rho-aminohippurate. These membranes did not exhibit Na+-dependent, phlorizin-inhibitable D-glucose transport. Sodium dodecyl sulfate polyacrylamide gel electrophoresis of proteins from P alpha revealed at least six major protein bands with molecular weights of 91000, 81000, 73000, 65000, 47000 and 38000. A small fraction of total alkaline phosphatase found in the homogenate was found in pellet P4. Membranes containing this alkaline phosphatase activity were distributed widely over the gradient, with peak activity found at a density of 1.141 g/cm3. In contrast, when brush borders were subjected to gradient centrifugation under the same conditions as P4, alkaline phosphatase was found in a narrow distribution, with peak activity at a density of 1.158 g/cm3. The principle subcellular localization of the alkaline phosphatase found in P4 could not be determined unambiguously from the data, but the activity did not seem to be primarily associated with classical brush borders. PMID- 6293561 TI - Inhibition of a voltage-dependent cation channel in sarcoplasmic reticulum vesicles by caesium studied by using a potential-sensitive cyanine dye. AB - The effect of caesium on the cation transport system in sarcoplasmic reticulum vesicles has been analysed kinetically through Tris+ influx. The Tris+ influx was measured by following the change in K+ diffusion potential due to the mutual diffusion between K+ and Tris+ in the presence of valiomycin using a potential probe; 3,3'-dipropylthiadicarbocyanine iodide. The main results were as follows. (1) Tris+ influx increased when membrane potential became inside-negative. This suggests that Tris+ permeates through the channel which has a voltage-dependent gate. (2) Cs+ reacted with the cation transport system only from the outside of the vesicle and inhibited Tris+ influx. The inhibition follows a single-site titration curve with a voltage-dependent dissociation constant of 18 mM at -60 mV. The inhibition can be explained by assuming that Cs+ binds to a site located about 45% of the way through the membrane from the outside of the vesicle in the open state of the channel. These results are in good agreement with those reported by Coronado and Miller (Coronado, R. and Miller, C. (1979) Nature 288, 495-497), which were gained electrically by using sarcoplasmic reticulum vesicles incorporated into an artificial planar phospholipid bilayer. PMID- 6293560 TI - Incorporation of Na+ - Ca2+ antiporter and of (Na+ + K+)-ATPase into liposomes and demonstration of their non-identity. AB - (Na+ + K+)-ATPase was isolated from the grey matter of brain and incorporated into liposomes. Most of the reconstituted enzyme was oriented 'inside-out' with respect to its in vivo orientation and externally added ATP promoted Na+ uptake that was inhibitable by internally trapped ouabain. Using the same proteoliposomes, an Na+ - Ca2+ exchange system was observed as indicated by the following pieces of evidence. (1) The Na+ gradient provided the only readily apparent driving force for acceleration of Ca2+ accumulation into proteoliposomes. (2) The antiporter was specific for Ca2+, high Mg2+ excess did not inhibit Ca2+ antiport. (3) The Na+ efflux was dependent on the extravesicular Ca2+ concentration. (4) The Na+ efflux was not inhibited by tetrodotoxin. The demonstrated Na+ - Ca2+ exchange could not be related to (Na+ + K+)-ATPase protein, since it was not purified with (Na+ + K+)-ATPase, as followed from transport studies with liposomes containing (Na+ + K+)-ATPase of different specific activity. The results strongly indicate that plasma membranes isolated from the grey matter of brain contain an Na+ - Ca2+ exchange system and that the proteoliposomes are suitable for further purification of the carrier molecule. PMID- 6293562 TI - Studies on ouabain-complexed (Na+ +K+)-ATPase carried out with vanadate. AB - Vanadate is able to promote the binding of ouabain to (Na+ +K+)-ATPase and it is shown that vanadate is trapped in the enzyme-ouabain complex. Also ouabain-bound enzyme, the formation of which was facilitated by (Mg2+ +Na+ +ATP) or (Mg2+ +Pi), is accessible to vanadate when washed free of competing ligands used for the promotion of ouabain binding. For vanadate binding to (Na+ +K+)-ATPase and to enzyme-ouabain complexes a divalent cation (Mg2+ or Mn2+) is indispensable, indicating that the cation does not remain attached to the ouabain-bound enzyme. K+ further increases vanadate binding in the absence of ouabain, but seems to have no additional role in case of vanadate binding to enzyme-ouabain complexes. Mn2+ is more efficient than Mg2+ in promoting binding of vanadate and ouabain to (Na+ +K+)-ATPase. That K+ in combination with Mn2+, in analogy with the effect in combination with Mg2+, increases the equilibrium binding level of vanadate and decreases that of ouabain does not seem to favour the hypothesis of selection of a special E2-subconformation by Mn2+. The vanadate-trapped enzyme-ouabain complex was examined for simultaneous nucleotide binding which could demonstrate a two substrate mechanism per functional unit of the enzyme. The acceleration by (Na+ +ATP) of ouabain release from the (Mg2+ +Pi)-facilitated enzyme-ouabain complex does not, as anticipated, support such a mechanism. On the other hand, the deceleration of vanadate release as well as of ouabain release from a (Mg2+ +vanadate)-promoted complex could be consistent with a two-substrate mechanism working out-of-phase. PMID- 6293564 TI - Quinidine is a strong perturber of acidic phospholipid bilayer order and fluidity. AB - The effect of an antiarrhythmic drug, quinidine, on the organization of model phospholipid membranes was studied by the spin-labeling technique. Quinidine strongly perturbs the molecular organization of lipid bilayers prepared from acidic phospholipids (phosphatidylserine, phosphatidic acid) and has only a slight effect on neutral phosphatidylcholine membranes. The interaction of the drug with acidic phospholipids manifests itself in a pronounced increase in the order parameter of the region close to the polar surface of the bilayer and in some decrease in its inner hydrocarbon core fluidity. It is suggested that the perturbation in the organization of membrane lipids may contribute to the mechanisms by which quinidine exerts its pharmacological effects. PMID- 6293563 TI - Detergent effects of kinetic properties of (Na+ +K+)-ATPase from kidney membranes. AB - Studies on (Na+ +K+)-ATPase generally employ detergents such as SDS and deoxycholate. Under such conditions, the purified enzyme possesses high specific activity. The (Na+ +K+)-ATPase from kidney membranes was unmasked by deoxycholate and SDS as described by Jlrgensen and its kinetic properties were studied. The results suggest that these detergents induce some irreversible alterations in the kinetic properties of the native enzyme. Another detergent, saponin, unmasked the (Na+ +K+)-ATPase as effectively as did SDS, but it seems to affect the kinetic properties of the native enzyme to a lesser extent. PMID- 6293565 TI - Effect of fatty acids on plasma membrane lipid dynamics and cation permeability in neuroblastoma cells. AB - In this study the effects of experimental modifications of plasma membrane lipid lateral mobility on the electrical membrane properties and cation transport of mouse neuroblastoma cells, clone Neuro-2A, have been studied. Short-term supplementation of a chemically defined growth medium with oleic acid or linoleic acid resulted in an increase in the lateral mobility of lipids as inferred from fluorescence recovery after photobleaching of the lipid probe 3,3' dioctadecylindocarbocyanide iodide. These changes were accompanied by a marked depolarization of the membrane potential from -51 mV to -36 mV, 1.5 h after addition, followed by a slow repolarization. Tracer flux studies, using 86Rb+ as a radioactive tracer for K+, demonstrated that the depolarization was not caused by changes in (Na+ + K+)-ATPase-mediated K+ influx or in the transmembrane K+ gradient. The permeability ratio (PNa/PK), determined from electrophysiological measurements, however, increased from 0.10 to 0.27 upon supplementation with oleic acid or linoleic acid. This transient rise of PNa/PK was shown by 24Na+ and 86Rb+ flux measurements to be due to both an increase of the Na+ permeability and a decrease of the K+ permeability. None of these effects occurred upon supplementation of the growth medium with stearic acid. PMID- 6293566 TI - Characterization of a beta-actinin-like protein in purified non-muscle cell membranes. Its activity on (Na+ + K+)-ATPase. AB - Treatment by EDTA of purified plasma membranes from MF2S cells (a variant of the murine plasmacytoma MOPC 173) solubilized proteins and increased by a 1000-fold the sensitivity of (Na+ + K+)-ATPase to ouabain. When added back with Ca2+ to treated plasma membranes, these EDTA-solubilized proteins restored the initial sensitivity of the enzyme to its inhibitor. We report the purification of a protein of Mr 32000, isolated from the EDTA-treated membrane supernatant. This protein was purified by a one-step procedure involving a preparative polyacrylamide gel electrophoresis without detergent. In the presence of Ca2+ it was able to restore the original sensitivity to ouabain of (Na+ + K+)-ATPase from EDTA-treated membrane. This protein was shown to be similar to the beta-actinin described by Maruyama by the following criteria: (1) molecular weight and amino acid composition; (2) cross-reactivity with their respective antisera; (3) in the presence of Ca2+ the same quantitative biological activity on ouabain sensitivity of the (Na+ + K+)-ATPase. A possible interaction between beta-actinin, calmodulin and membrane-bound (Na+ + K+)-ATPase is discussed. PMID- 6293567 TI - Inhibition of gastric (H+ + K+)-ATPase by unsaturated long-chain fatty acids. AB - Arachidonic acid and unsaturated C18 fatty acids at concentrations near 10(-5) M markedly inhibited (H+ + K+)-ATPase in hog or rat gastric membranes. Arachidonic acid was a more potent inhibitor than unsaturated C18 fatty acids, but the involvement of the metabolites of arachidonic acid cascade was ruled out. Linolenic acid inhibited the formation of phosphoenzyme and the K+ -dependent p nitrophenylphosphatase activity of the hog ATPase. Treatment with fatty acid-free bovine serum albumin abolished only the inhibitory effect of the fatty acid on the phosphatase activity without restoring the overall ATPase action. These data suggest the existence of at least two groups of hydrophobic binding sites in the gastric ATPase for unsaturated long-chain fatty acids which affect differentially the catalytic reactions of the ATPase. (H+ + K+)-ATPase in rat gastric membranes was found more susceptible to the fatty acid inhibition and also more unstable than the ATPase in hog gastric membranes. The presence of a millimolar level of lanthanum chloride or ethylene glycol bis(beta-aminoethyl ether)-N,N'-tetraacetic acid stabilized the rat ATPase probably via the inhibition of Ca2+ -dependent phospholipases in the gastric membranes. PMID- 6293568 TI - Topology of membrane exposure in the renal cortex slice. Studies of glutathione and maltose cleavage. AB - We measured glycine release from ([2-3H]glycine)-labelled GSH and glucose formation from maltose incubated with rat kidney whole cortex homogenate, thin cortex slices or collagenase-treated tubule fragments. Liberation of glycine was inhibited (74-83%) by serine borate (20 mM), indicating a gamma glutamyltransferase-dependent hydrolysis of GSH. In whole cortex homogenate, the GSH cleavage activity was 17.4 +/- 0.6 nmol GSH degraded/mg protein per min (mean +/- S.D.); cleavage activity by intact slices was 3.5 +/- 0.7 (P less than 0.001 relative to whole cortex homogenate) and in tubule fragments 9.4 +/- 0.8 (P less than 0.001). Homogenizing the tissue preparation increased cleavage rate in slices about 4-fold (12.4 +/- 2.9; P less than 0.005 relative to intact slice) but did not change the rate in tubule fragments (9.8 +/- 0.5). Maltose cleavage activity in whole cortex homogenate was 512 +/- 22 nmol glucose formed/mg protein per min, in slices 162 +/- 12, and in tubules 884 +/- 48. These findings imply that substrate in the incubation medium has a limited access to the luminal membrane of cortex slices but not of tubule fragments. They further imply that basolateral membrane is preferentially exposed in the slice preparation. PMID- 6293569 TI - Possible role of non-bilayer lipids in the structure of mitochondria. A freeze fracture electron microscopy study. AB - The possible role of non-bilayer phospholipids on the structure of isolated rat liver mitochondria has been morphologically studied. Freshly isolated freeze fractured mitochondria show smooth fracture faces with particles, representing the limiting membranes. The frequency and size of the particles is representative for the various membrane faces. Distinctly large particles and pits represent the attachment sites of cristae to the inner membrane. Liposome-like structures in the matrix are found upon incubation with Ca2+ and Mn2+. At 5 mM Mn2+ and more, curved hexagonal (HII) phase tubes are observed. Subsequent addition of 1 mM EDTA results in disappearance of the HII tubes, and liposomal structures can again be seen. These findings are interpreted in terms of an Mn2+-induced lamellar to HII phase transition. Patchwork-like structures characterize the membranes of mitochondria, quenched from 37 degrees C, as well as those incubated with Ca2+, Mn2+, Mg2+ and apo- or cytochrome c. This phenomenon is interpreted as being the result of the fracture plane, jumping from the outer to the inner limiting membrane and vice versa at sites of contact. A semi-fusion model, in which non bilayer lipids are involved, is proposed for these contact sites. PMID- 6293570 TI - Uniform ionophore A23187 distribution and cytoplasmic calcium buffering in intact human red cells. AB - The divalent cation-selective ionophore A23187 has been used to characterize cytoplasmic Ca and Mg buffering, Ca2+-pump parameters and the properties of a Ca2+-activated K+-channel in intact red cells. A critical assumption in these studies has been that the ionophore causes a uniform increase in divalent cation permeability in all the cells. This has now been tested directly in ATP-depleted human red cells by analysing the kinetics of ionophore-induced 45Ca-tracer and net Ca2+ fluxes. The experimental curves were all adequately fitted by single exponentials at all ionophore concentrations tested. Moreover, statistical analysis of 61 individual tracer influx curves and of pooled data showed no trend towards fast second exponential components. These results demonstrate uniformity of ionophore distribution, ionophore-induced Ca2+-permeability, and cytoplasmic Ca-buffering among all the cells. Experiments involving mixing of cell suspensions with high and low original ionophore content, and involving ionophore extraction by albumin, demonstrate a rapid redistribution of ionophore among the cells, indicating that homogeneity of ionophoric effects is achieved through dynamic ionophore redistribution. PMID- 6293571 TI - Orientation of rat-liver plasma membrane vesicles. A biochemical and ultrastructural study. AB - Using both biochemical and morphological methods, the membrane orientation of plasma membrane vesicles from rat liver which are capable of catalysing the active transport of amino acids was investigated. In intact vesicles, the plasma membrane enzyme (Na+ + K+)-ATPase displays only a minor portion of its total activity which is greatly increased upon vesicle disruption. The same intact vesicles show an almost maximal binding of ouabain, which binds only to the extracellular side of the plasma membrane. A freeze-fracture analysis of the vesicles shows that a distinct population of relatively large vesicles have predominantly the in vivo membrane orientation. These large vesicles are labelled with numerous filipin-sterol complexes following exposure to the cholesterol probe, filipin, and are therefore assumed to be plasma membrane vesicles. A population of smaller vesicles with mainly an inside-out orientation were not labelled with filipin and are probably microsomes. The data obtained with both biochemical and ultrastructural techniques indicate that the plasma membrane vesicles isolated from rat liver for transport studies are mostly (at least 70%) orientated as in vivo, i.e. inside-in. PMID- 6293572 TI - A fluctuation analysis study of the development of amiloride-sensitive Na+ transport in the skin of larval bullfrogs (Rana catesbeiana). AB - In the presence of the Na+ -channel blocker amiloride, the short-circuit current across the skins of bullfrog tadpoles in metamorphic stages XIX-XXIV was subjected to fluctuation analysis. The resulting power spectra contained a Lorentzian component of which the plateau value (S0) decreased while the corner frequency (fc) increased as the mucosal amiloride concentration was increased from 0.5 to 24 microM. From the linear relationship between the fc values and the amiloride concentrations it was possible to determine the binding (k'01) and unbinding (k10) constants for amiloride to its receptor on the Na+ channel. With these parameters as well as short-circuit current and S0 values, the current through the individual Na+ channels (i) was calculated (average 0.58 pA). It did not increase significantly during late metamorphosis. The density of Na+ channels (M) in the apical membrane, on the other hand, increased significantly. It would appear that the increase in short-circuit current which occurs at this time is due primarily to an increase in amiloride-blockable Na+ channels. Unexpectedly, a Lorentzian component could be fitted to power spectra in amiloride-treated skins (stages XIX-XXI) which showed no amiloride-sensitive short-circuit current. Moreover, the typical increase in fc with the amiloride concentration did not occur in these animals. PMID- 6293573 TI - beta-Adrenergic receptors of brain cells. Membrane integrity implies apparent positive cooperativity and higher affinity. AB - Beta-Adrenergic receptors were studied in intact cells of chick, rat and mouse embryo brain in primary cultures, by the specific binding of [3H]dihydro-L alprenolol ([3H]DHA). The results were compared to the receptor binding of broken cell preparations derived from the cell cultures or from the forebrain tissues used for the preparation of the cultures. Detailed analysis of [3H]DHA binding to living chick brain cells revealed a high-affinity, stereoselective, beta adrenergic-type binding site. Equilibrium measurements indicated the apparent positive cooperativity of the binding reaction. By direct fitting of the Hill equation to the measured data, values of Bmax = 12.01 fmol/10(6) cells (7200 sites/cell), Kd = 60.23 pM and the Hill coefficient n = 2.78 were found. The apparent cooperative character of the binding was confirmed by the kinetics of competition with L-alprenolol, resulting in maximum curves at low ligand concentrations. The rate constants of the binding reaction were estimated as k+ = 8.31 X 10(7) M-1 X min-1 and k- = 0.28 min-1 from the association results, and k- = 0.24 min-1 from the dissociation data. The association kinetics supported the cooperativity of the binding, providing a Hill coefficient n = 1.76; Kd, as (k /k+)1/n was found to be 101 pM. Analysis of the equilibrium binding of [3H]DHA to rat and mouse living brain cells resulted in values of Bmax = 13.04 fmol/10(6) cells (7800 sites/cell), Kd = 43.85 pM and n = 2.52, and Bmax = 8.08 fmol/10(6) cells (4800 sites/cell), Kd = 46.70 pM and n = 1.63, respectively, confirming the apparent cooperativity of the beta-receptor in mammalian objects, too. The [3H]DHA equilibrium binding to broken cell preparations of either chick, rat or mouse brain cultures or forebrain tissues was found to be non-cooperative, with a Hill coefficient n = 1, Kd in the range 1-2 nM, and a Bmax of 10(3) - 10(4) sites/cell. Our findings demonstrate that cell disruption causes marked changes in the kinetics of the beta-receptor binding and in the affinity of the binding site, although the number of receptors remains unchanged. PMID- 6293574 TI - The effect of ACTH on rat brain synaptic plasma membrane lipid fluidity. AB - The effect of ACTH on the lipid fluidity was examined in synaptic plasma membranes from rat forebrain. ACTH1-24 increased the fluidity of the synaptic plasma membranes in a dose-dependent way, the lowest effective dose being 10(-5) M. The shorter N-terminal fragment ACTH1-10 was not effective. The significance of this finding is discussed in relation to the known effects of ACTH on synaptic membrane phosphorylation. PMID- 6293575 TI - Inhibition of protein synthesis by a tryptic polypeptide of Clostridium perfringens type A enterotoxin. AB - The biological activity of Clostridium perfringens enterotoxin can be tested more precisely and with a much higher sensitivity by using the inhibition of protein synthesis by Vero cells, rather than the guinea pig skin test. Tryptic peptides of the enterotoxin produced in the presence of different concentrations of sodium dodecyl sulfate (0-1%) have been tested for biological activity (Vero cells) and inhibitory effect on cell-free protein synthesis (rabbit reticulocyte lysate). A fraction of tryptic peptides, about 16,000 daltons, was able to inhibit the cell free protein synthesis, while the native enterotoxin had no such effect. The 16 kDa fraction had, however, lost the ability to disrupt the Vero cells (normal biological activity). It is probable that the enterotoxin has the double function (A and B chain), known from several other toxins, confined in its single polypeptide chain. PMID- 6293576 TI - Adenosine diphosphate: thymidine 5'-phosphotransferase, a new enzyme activity, associated with the Herpes simplex virus-induced deoxypyrimidine kinase. AB - The deoxypyrimidine kinase induced in mouse fibroblasts, strain CLID (TK-) infected with either herpes simplex virus (HSV) type 1 or type 2, possesses besides deoxypyrimidine kinase (ATP:dThd/dCyd phosphotransferase) two further enzyme activities: an AMP:dThd phosphotransferase and an ADP:dThd phosphotransferase. The latter enzyme activity, described in this report, was found to be inhibited by antiserum against the HSV deoxypyrimidine kinase and to be absent after infection with TK- mutant MDK 10 (B 2006). The ADP:dThd phosphotransferase, which had been purified approx. 340-fold, differs by a series of physicochemical properties from the viral AMP:dThd- and ATP:dThd phosphotransferase. PMID- 6293577 TI - Activation of phosphodiesterase in frog rod outer segment by rhodopsin analogues. AB - Activation of guanosine 3',5'-cyclic monophosphate (cGMP) phosphodiesterase (EC 3.1.4.35) in frog rod outer segment membrane by rhodopsin analogues has been investigated. A rhodopsin analogue modified at the Schiff-base linkage (N-retinyl opsin) or the beta-ionone ring (3-dehydro-rhodopsin) in the retinylidene chromophore of rhodopsin has some ability in activation of the enzyme. In consideration of our previous observation that opsin including a retinal-oxime can activate the enzyme, it seems likely that the Schiff-base linkage is not always necessary for the phosphodiesterase activation. On the other hand, a change in the length of the side chain of retinal (complex of opsin and beta ionone, beta-ionylideneacetaldehyde or retinylideneacetaldehyde) or dissection of the conjugate double-bond system of the side chain (retro-gamma-rhodopsin) remarkably reduces the activation ability. However, 5,8-epoxy-rhodopsin having a similar dissected conjugate double-bond system induces some enzyme activation because of its rigid conformation around C7-C8-C9 single bonds. Consequently, it is suggested that the necessary portion of rhodopsin chromophore for the activation of the enzyme is the rigid conjugate double-bond system between the beta-ionone ring and the Schiff-base linkage in its all-trans form. PMID- 6293578 TI - Lysosomal and microsomal beta-glucuronidase of monkey brain. Differential elution characteristics from con A-sepharose and neutral sugar composition. AB - Microsomal and lysosomal beta-glucuronidase (beta-D-glucuronide glucuronosohydrolase, EC 3.2.1.31) of monkey brain were differentially eluted from Con A-Sepharose when subjected to chromatography and linear gradient elution with methyl alpha-glucoside at 28+/-1 degree C. The lysosomal enzyme was eluted as a sharp peak in the first few fractions, while the microsomal enzyme was eluted as a broad peak extending over several fractions. This differential pattern of elution was dependent only on the temperature of elution and the concentration of methyl alpha-glucoside used. The lysosomal and microsomal glucuronidases were purified to apparent homogeneity and their neutral sugar analysed. Both of them contained glucose, mannose and fucose but the microsomal enzyme contained about 3-times as much of all these sugars as the lysosomal enzyme. Sodium periodate treatment of the microsomal enzyme resulted in a shift in its elution pattern, similar to the lysosomal enzyme when subjected to Con A Sepharose chromatography. The content of neutral sugars and the structural features of the oligosaccharide units in the microsomal glucuronidase might be responsible for its elution pattern. A processing of the carbohydrate units of the microsomal glucuronidase might be envisaged to take place if it were to act as a precursor of the lysosomal glucuronidase. PMID- 6293579 TI - Collagenase digestion demonstrates carboxy-terminal crosslinking in acid-soluble collagen. AB - Among the products of the collagenase cleavage of Type I acid-soluble collagen from calf and rabbit tendons, there can be found fragments with the lengths of half alpha-chains. Because purified collagenase cleaves the alpha-chains three quarters of the length from the amino-terminus, the presence of half-length chains is evidence for the occurrence of crosslinks between two carboxy-terminal, quarter-length fragments, The collagen preparations were reduced with [3H]borohydride, the collagenase-cleaved fragments were separated by gel electrophoresis, and their 3H-labeled crosslink derivatives were analyzed. The major labeled components in the half-length chains were the reduced aldol condensation product and hydroxylysinonorleucine. These experiments demonstrate that the carboxy-terminal telopeptides in monomer-enriched collagen samples form aldol crosslinks which are probably intramolecular, but some intermolecular aldol and aldimine crosslinks may also be formed. PMID- 6293580 TI - Purification and characterization of phosphoglycerate mutase isozymes from pig heart. AB - The three isozymes of phosphoglycerate mutase from pig heart have been purified to homogeneity. The isozymes have a molecular weight of 57000 as determined by gel-filtration chromatography. Discontinuous gel electrophoresis in the presence of sodium dodecyl sulfate yields a single band with a molecular weight of 29000, indicating that the isozymes are dimers composed of subunits of similar mass. Hybridization experiments show that the three isozymes result from homodimeric and heterodimeric combinations of two different subunits. The two types of subunit differ in their heat lability and in the presence of -SH groups essential for enzymatic activity. No remarkable differences exist in the kinetic constants of the purified isozymes. The kinetic pattern is consistent with a 'ping-pong' mechanism. The homogeneous preparations of the three isozymes show intrinsic glycerate-2,3-P2 synthase activity and glycerate-2,3-P2 phosphatase activity which can be stimulated by glycolate-2-P. PMID- 6293581 TI - Comparison of some physicochemical and kinetic properties of S adenosylhomocysteine hydrolase from bovine liver, bovine adrenal cortex and mouse liver. AB - S-Adenosyl-L-homocysteine hydrolase (EC 3.3.1.1) was purified to apparent homogeneity from bovine liver, bovine adrenal cortex and mouse liver. All enzymes were tetramers, composed of two types of subunit present in the proportion 1:1, as judged by SDS-polyacrylamide gel electrophoresis. The partition coefficient was exactly the same for these enzymes on high-performance gel permeation chromatography, and they co-sedimented in density gradients, suggesting the same molecular size and form of S-adenosylhomocysteine hydrolase from these sources. The bovine enzymes differed from the mouse liver enzyme with respect to isoelectric point (pI = 5.35, versus pI = 5.7), affinity for DEAE-cellulose, and migration of subunits on SDS-polyacrylamide gel electrophoresis with SDS from some commercial sources. The enzymes were not substrates for cAMP-dependent protein kinase. The apparent Km values for adenosine (0.2 microM) and S adenosylhomocysteine (0.75 microM) were the same for all three enzymes. The ratio between Vmax for the synthesis and hydrolysis of S-adenosylhomocysteine was about 4 for the mouse liver enzyme, and about 6 for the bovine enzymes. It is concluded that only subtle kinetic and physicochemical differences exist between S adenosylhomocysteine hydrolase from these bovine and mouse tissues. This suggests that differences in experimental procedures rather than species- and organ differences of S-adenosylhomocysteine hydrolase are responsible for the variability in kinetic and physicochemical parameters reported for the mammalian hydrolase. PMID- 6293582 TI - Axial histidyl imidazole non-exchangeable proton resonances as indicators of imidazole hydrogen bonding in ferric cyanide complexes of heme peroxidases. AB - Proton NMR spectra of a model of low-spin cyanide complexes of ferric hemoproteins indicate that two broad single-protein resonances from the axial imidazole can be resolved outside the diamagnetic spectral region. Upon deprotonation of the imidazole in the model, the upfield resonance shifts dramatically to higher field, suggesting that its position may reflect the degree of hydrogen bonding or proton donation of the imidazole. Met-cyano myoglobin reveals a pair of such broad peaks in the regions expected for an essentially neutral axial imidazole. In the cyano complexes of horseradish peroxidase and cytochrome c peroxidase, a pair of single-proton resonances are located which are assigned to the same imidazole protons on the basis of their linewidth and shift changes upon altering the heme substituents. The upfiled proton, however, is found at much higher field than in metMbCN. The upfield bias of this resonance is taken as evidence for appreciable imidazolate character for the axial ligand in these heme peroxidases. PMID- 6293583 TI - Cytochrome c oxidase from the liver of bullfrog, Rana catesbeiana and change in its turnover rate during metamorphosis. AB - 1. Cytochrome c oxidase was purified from the liver mitochondria of bullfrog (Rana catesbeiana). The heme a content of the purified enzyme was 13.5 nmol per mg protein. Polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate revealed that the enzyme protein was composed of nine polypeptide subunits having molecular weights of 42000, 27000, 25000, 20000, 15000, 13000, 8600, 5400 and 3600. The purified enzyme from the adult frog was immunological identified with that from the tadpole. 2. The ratio of synthesis and degradation of cytochrome c oxidase were 5.2- and 2.0-times higher at metamorphic climax than at premetamorphic stage, respectively. The amount of the enzyme in the liver was highest at metamorphic climax. PMID- 6293584 TI - Subcellular localization and substrate specificity of dolichol kinase from rat liver. AB - When purified subcellular fractions were prepared from rat liver and assayed for dolichol kinase activity using pig liver dolichol as a substrate, the microsomes were found to contain the highest specific activity and greater than 75% of the total activity. With regard to substrate specificity, the microsomal enzyme showed a marked preference for saturation of the alpha-isoprene: dolichol-16 and 19 were 2.5-fold more active than the corresponding polyprenols. For a given class of prenol, the 16 and 19 isoprenologs exhibited similar activity, whereas the 11 isoprenolog appeared less active. The enzyme was twice as active against the naturally occurring polyprenol-16 (alpha-cis-isoprene) compared to synthetic alpha-trans-polyprenol-16. Taken together, the data indicate that the alpha isoprene specificity follows the order: saturated greater than cis greater than trans. In addition, all-trans-2,3-dihydrosolanesol was not a substrate, suggesting that at least one cis isoprene residue is required. PMID- 6293585 TI - Localization of neutral magnesium-stimulated sphingomyelinase in plasma membrane of cultured neuroblastoma cells. PMID- 6293586 TI - The production of free radicals during the autoxidation of cysteine and their effect on isolated rat hepatocytes. AB - Autoxidizing cysteine has been shown to produce thiyl and hydroxyl radicals. Hydrogen peroxide increased the yield of both radicals which was inhibited by catalase but stimulated by copper/zinc superoxide dismutase. This effect is due to increased hydrogen peroxide production by copper/zinc superoxide dismutase as a result of superoxide dismutation. The production of superoxide radicals could not be detected probably because of its low reactivity, however, measurement of oxygen uptake and reduction of ferricytochrome c by autoxidizing cysteine clearly implicate the involvement of super oxide radicals. The production of hydroxyl radicals is postulated to proceed through a fenton reaction, however, this may not necessarily be metal ion controlled. Autoxidizing cysteine disrupts the integrity of hepatocytes causing release of glutathione, adenosine triphosphate and lactate dehydrogenase indicating that it is of little use as a therapeutic agent. PMID- 6293587 TI - Volume regulation of human peripheral blood lymphocytes and stimulated proliferation of volume-adapted cells. AB - Human peripheral blood lymphocytes exposed to hypotonic media (Ca/Mg-free, room temp.) first swell and then shrink. This shrinking response is characterized by a simple exponential with a half-time of 1.44 +/- 0.60 min (n = 11) and its extent but not the half-time for a given hypotonicity is influenced by [K+]0. Using K selective electrodes, we observe a change in [K+]0 when cells are diluted into hypotonic media. A half-time of 1.55 +/- 0.06 min (n = 4) was obtained. A similar half-time was obtained by assay of total cell K using atomic absorption spectroscopy. At all osmolarities [K+]i was decreased from control values and was constant as [K+]0 was increased. Short-term incubation with ouabain (10(-4) M) had no effect. Decreasing osmolarities progressively inhibited phytohemagglutinin stimulated DNA synthesis, yet cell number and viability remained unaltered. Our evidence indicates that the volume response is mediated by a change in the passive permeability of the plasma membrane to K and/or to the accompanying anions, and that the consequently volume-adapted cells are growth-inhibited. PMID- 6293588 TI - Effects of sulfonylureas on membrane-bound low Km cyclic AMP phosphodiesterase in rat fat cells. AB - The effects of sulfonylureas and a biguanide on membrane-bound low Km cyclic AMP phosphodiesterase and lipolysis were examined in rat fat cells. Pharmacologically active sulfonylureas, such as tolbutamide (10 mM), acetohexamide (10 mM) and glibenclamide (200 microM) activated the phosphodiesterase when incubated with fat cells and suppressed lipolysis induced by isoproterenol. However, neither of these actions was observed in the presence of a pharmacologically inactive sulfonylurea, carboxytolbutamide (10 mM) and a biguanide, buformin (500 microM). Tolbutamide (0.5-10 mM) activated the enzyme, concentration dependently, and this manner of activation appears to coincide with that of the suppressive effect on the lipolysis. The time course of the enzyme activation was similar to that seen with insulin. Km, optimal pH and sensitivity to temperature of the enzyme from tolbutamide-treated cells were the same as those of the enzyme from control and insulin-treated cells. Direct incubation of the enzyme from control cells with tolbutamide did not affect the activity, while as little as 10 microM 3-isobutyl 1-methylxanthine markedly inhibited the enzyme. Tolbutamide continued to activate the enzyme in cells in which insulin receptor had been destroyed by trypsin pretreatment. These results are compatible with the idea that the enzyme activated by sulfonylurea and that activated by insulin may be the same species of phosphodiesterase and that the antilipolytic action of sulfonylurea may be mediated by the activation of the enzyme which does not occur through the insulin receptor. PMID- 6293589 TI - [Functional properties of the catecholamine-sensitive adenyl cyclase system in embryonic skeletal muscle]. AB - During embryogenesis the adenylate cyclase from chicken skeletal muscle possesses a high catalytic activity in comparison with the mature tissue. The embryonic muscle enzyme is sensitive to the stimulating effects of fluoride, vanadate and guanyl nucleotides. The enzyme sensitivity to catecholamines appears at the end of the 2nd week of embryogenesis. Activation of chicken skeletal muscle adenylate cyclase by catecholamines occurs via the adrenoreceptor of the beta-type. Upon combined action of guanyl nucleotides and adrenaline the potentiation effect typical for the mature muscle is observed only at post-embryonic stages. It is assumed that the functional development of the catecholamine--sensitive adenylate cyclase system in embryogenesis occurs in the following order: catalytic, regulatory, receptor components. The process is completed at the post-embryonic stage. PMID- 6293590 TI - [Binding of low density lipoproteins (LDL) to platelets]. AB - Binding of LDL to platelets was studied by two independent methods, radioactive and flow cytofluorimetry, using 125I- and fluorescently labelled RITC-LDL. Saturation of 125I- and RITC-LDL binding to platelets, inhibition of binding by unlabelled LDL and a lower inhibitory effect of unlabelled HDL evidence the existence of a limited number of binding sites specific for LDL in platelets. Unlike nuclear cells platelets do not degrade LDL. The binding of LDL to platelets is reversible and independent of Ca2+. The decrease of total binding level at 4 degrees and the absence of heparin effect on the release of bound LDL suggest LDL incorporation into platelets. PMID- 6293591 TI - [Generation of the differences of the electric potentials by Rhodospirillum rubrum reaction center complexes devoid of the heavy subunit]. AB - The electrogenic activity of Rhodospirillum rubrum P870 reaction center complexes devoid of the heavy (H) subunit and retaining the light (L) and medium (M) subunits, was studied. The proteoliposomes containing such reaction center complexes were formed by a self-assembly procedure, using soya bean phospholipids. In the presence of Ca2+ the reaction center proteoliposomes were incorporated into a phospholipid-impregnated Teflon filter separating two solutions of identical composition. After addition of N,N,N',N'-tetra-methyl-p phenylenediamine (or cytochrome c) and qinone (menadione), illumination caused generation of an electric potential difference between the two filter-separated compartments, the proteoliposome-free compartment being negatively charged. The illuminated proteoliposomes took up penetrating tetraphenylphosphonium cations and, in a lesser degree, tetraphenylborate anions. The data obtained suggest that the reaction center complexes containing only L- and M-subunits possess the electrogenic activity. The H-subunit is not directly involved in membrane potential generation. PMID- 6293592 TI - [Obtaining homo- and heterokaryons between mouse fibroblasts and rat peritoneal leukocytes and mastocytes using Sendai virus and polyethylene glycol]. PMID- 6293593 TI - Subsensitivity of catecholaminergic neurons to direct acting agonists after single or repeated electroconvulsive shock. AB - Spontaneous firing rates and changes in firing rate in response to an intravenously administered dose of apomorphine were measured after various electroconvulsive shock (ECS) treatment regimens from dopaminergic cells of the substantia nigra in urethane-anesthetized rats. Similar measurements were obtained from noradrenergic neurons of the locus coeruleus before and after intravenous injection of clonidine. A significant decrement in the inhibition of spontaneous firing in response to intravenous administration of these agonists was observed following multiple or single ECS treatment in both substantia nigra and locus coeruleus cells. There was a consistent but nonsignificant tendency for cells in both areas of the brain from treated animals to display higher rates of spontaneous firing than their respective sham-shocked controls. Both the effects on base-line rates of spontaneous activity and on the depression of firing rate in response to drug administration were found to be independent of repeated treatment. A significant negative correlation was obtained between base-line firing rate and percentage depression to the autoreceptor agonist, but this correlation alone was insufficient to account for the observed differences in the drug response. These results are discussed with respect to possible mechanisms of action of electroconvulsive therapy in the treatment of depression. PMID- 6293594 TI - beta-Endorphin hypersecretion in depression: possible cholinergic mechanisms. AB - Morning plasma concentrations of beta-endorphin immunoreactivity were significantly higher in a group of depressed patients meeting the Research Diagnostic Criteria for Major Depressive disorder or Schizo-affective disorder, depressed, than in age- and sex-matched groups of normal controls and psychiatric patients without affective disorders. Furthermore, physostigmine-stimulated release of beta-endorphin immunoreactivity was also significantly greater in the depressed patients. These results provide the first evidence for elevated plasma concentrations of beta-endorphin in depression and also represent further evidence for cholinergic supersensitivity in depression. These results suggest that elevated plasma concentrations of beta-endorphin and cholinergically stimulated hypothalamic-pituitary beta-endorphin release, might potentially represent biological state or trait markers for depression. PMID- 6293595 TI - Survival of the K+ channel in axons externally and internally perfused with K+ free media. AB - In perfused squid giant axons, potassium channels irreversibly deteriorate when the internal K+ is removed and replaced by impermeant ions. Under the same conditions in perfused Myxicola giant axons, the K+ conductance is also irreversibly lost with a time constant of 10-15 min. In contrast, the K+ conductance in Myxicola giant axons dialyzed with impermeant ions and bathed in monovalent cation free solutions does not deteriorate, even over 5-6 h. Thus we suggest that washout of some internal component may be an important additional factor in the deterioration of K+ channels in perfused giant axons. PMID- 6293596 TI - Simultaneous modifications of sodium channel gating by two scorpion toxins. AB - The effects of purified scorpion toxins from two different species on the kinetics of sodium currents were evaluated in amphibian myelinated nerves under voltage clamp. A toxin from Leiurus quinquestriatus slowed and prevented sodium channel inactivation, exclusively, and a toxin from Centruroides sculpturatus Ewing reduced transient sodium currents during a maintained depolarization, and induced a novel inward current that appeared following repolarization, as previously reported by Cahalan (1975, J. Physiol. [Lond.]. 244:511-534) for the crude scorpion venom. Both of these effects were observed in fibers treated with both of these toxins, and the kinetics of the induced current were modified in a way that showed that the same sodium channels were modified simultaneously by both toxins. Although the toxins can act on different sites, the time course of the action of C. sculpturatus toxin was accelerated in the presence of the L. quinquestriatus toxin, indicating some form of interaction between the two toxin binding sites. PMID- 6293597 TI - Jumping frequencies in membrane channels. Comparison between stochastic molecular dynamics simulation and rate theory. AB - Permeation of molecules through membrane channels involves local interactions with a limited number of ligand groups. A method for the molecular dynamics simulation of particle movement in small ligand systems is described. It is assumed that the ligand groups carry out thermal vibrations, whereas the rest of the channel molecule and the surroundings act as a heat bath which is coupled via random forces to the motions of the ligands. The simulation technique is applied to a simple system which contains some of the essential features influencing jumping rates in membrane channels, such as flexibility of ligand configuration or inertial effects in the motion of the ligands. Since the simulation is based on strictly microscopic parameters of the particle-ligand system, a rigorous test of the predictions of rate theory is possible. It is found that rate theory describes the general dependence of jumping frequency k' on temperature and on ligand binding strength rather well, although the values of k' obtained by computer simulation are 2-3 times smaller than those predicted by rate theory. PMID- 6293598 TI - The gating of single calcium-dependent potassium channels is described by an activation/blockade mechanism. AB - Single calcium dependent potassium channels from cultured rat myoballs have been studied with the patch clamp technique, and current records subjected to statistical analysis. From the dependence of the mean open state probability on the internal calcium concentration, two calcium ions are required to open the channel. The open state and closed state lifetime distributions reveal that the usual activation model is not applicable to these channels. They are consistent with a two step gating mechanism that involves both activation by calcium and blockade by a calcium-sensitive gate. PMID- 6293599 TI - Molecular dynamics of ferrocytochrome c: anharmonicity of atomic displacements. PMID- 6293600 TI - Localized energy coupling during photophosphorylation by chromatophores of Rhodopseudomonas capsulata N22. AB - The principle of the dual inhibitor titration method for testing models of electron-transport phosphorylation is outlined, and the method is applied to the study of photophosphorylation in bacterial chromatophores. It is concluded that energy coupling is strictly localized in nature in this system, in the sense that free energy released by a particular electron-transport chain may be used only by a particular H+-ATP synthase. Dual inhibitor titrations using the uncoupler SF 6847 and the H+-ATP synthase inhibitor oligomycin indicate that uncouplers act by shuttling rapidly between the localized energy-coupling sites. PMID- 6293601 TI - The effects of S-adenosylhomocysteine and S-adenosylmethionine on some purine- and pyrimidine-metabolizing systems. AB - The effects of S-adenosylhomocysteine and S-adenosylmethionine on some purine- and pyrimidine-metabolizing systems have been examined. Both compounds were capable of acting as relatively good inhibitors of adenosine deaminase, nucleoside phosphorylase, and adenylate deaminase activities but as relatively poor inhibitors of myokinase and nucleoside monophosphate kinase. The inhibitory effects were freely reversible. 5'-Nucleotidase, orotidine 5'- phosphate, and phosphodiesterase were unaffected. Nucleoside phosphorylase was competitively inhibited by both compounds, whereas mixed inhibitory effects occurred with adenosine deaminase. PMID- 6293602 TI - Hereditary retinal degenerations in the dog: specificity of abnormal cyclic nucleotide metabolism to diseases of arrested photoreceptor development. PMID- 6293603 TI - The significance of genetic research in ophthalmology. PMID- 6293604 TI - Lipid inclusions in human ocular tissues in vitro induced by aminoglycoside antibiotics. PMID- 6293606 TI - [Possible localization of the gene of sensitivity of human cells to Coxsackie B virus in the short arm of chromosome 21]. PMID- 6293605 TI - Current concept of macular corneal dystrophy. PMID- 6293607 TI - [Affinity of various antidepressive agents to imipramine receptors of synaptic membranes in mouse brain]. PMID- 6293608 TI - [Effect of diazepam on reactivity of hippocampal neurons during blocking of GABA ergic system]. AB - It was suggested that diazepam action is mediated by the GABA-ergic system. This suggestion was controlled in vitro experiments with hippocampal slices. The GABA ergic system was blocked by bicuculline and picrotoxin. The influence of diazepam (1 X 10(-6) M) on the population spikes (PS) evoked in the CA1 area by Schaffer's collaterals stimulation manifested itself in a reversible decrease. Bicuculline (1 X 10(-7) M) and picrotoxin (1 X 10(-6) M) induced an increase in the PS amplitude and appearance of additional PS. Diazepam was normally ineffective when applied in conjunction with bicuculline or picrotoxin. The data indicate that the action of diazepam is really mediated by the GABA-ergic system. PMID- 6293609 TI - [Inactivation of fast sodium current across the membrane of isolated cardiocytes]. AB - Inactivation of sodium conduction across the membrane of rat single heart cells was studied by the patch-voltage-clamp method. The development of sodium channel inactivation was found to be double-exponential. Dependence of both time constants on the membrane potential was measured. The time course of the recovery of the sodium conduction from inactivation was sigmoidal in shape with marked retardation at the beginning. The data obtained are not covered by the Hodgkin Huxley formalismus. PMID- 6293610 TI - [Identification and evaluation of properties of benzodiazepine receptors of glial and neuronal fractions of human cerebral cortex]. PMID- 6293612 TI - [Characteristics of GABA-potentiating effect of harman]. AB - Experiments on an isolated spinal cord of rats aged 9-15 days have shown that harmane (10(-7)-10(-5) M) enhances GABA-induced (1.10(-4) M) depolarization of primary afferents and hyperpolarization of motoneurones. The GABA-potentiating action of harmane on primary afferents is depicted by a bell-shaped curve with a maximum at 10(-5) M. The action is more pronounced the higher the concentration of chlorine ions in the medium. Harmane (10(-6)-10(-4) M) enhances spontaneous neuronal activity and evoked synaptic potentials (mono- and polysynaptic potentials of the ventral and dorsal roots) at the concentrations at which it exerts a direct depolarizing action on motoneurones and primary afferents. At higher concentrations the stimulant activity of harmane (10(-5)-10(-4) M) counteracts its GABA-potentiating effects. PMID- 6293611 TI - [Nature of potentiation of effects of gamma-aminobutyric acid (GABA) by benzodiazepine tranquilizers]. PMID- 6293613 TI - [Effect of diethylamine analog of ethmozine on parameters of fast sodium current in normal and depolarized myocardial fibers]. AB - The effect of a diethylamine analog of ethmozine (DAAE) on fast sodium current of normal and depolarized frog atrial trabeculae was studied by means of the double sucrose gap technique. The depolarization of the fibers was produced both by increasing extracellular potassium concentration up to 8 +/- 9 mM and by current passing. The resting potential of normal fibers was within the range of 75-80 mV, and the depolarized one was within 65 +/- 60 mV. DAAE (8 X 10(-7) g/ml) reduced sodium conductivity, slowed inactivation and reactivation of fast sodium current and shifted the steady-state activation curve (h infinity) to a more negative potential, but the steady-state activation curve (m infinity) to a more positive potential. All these effects were expressed more considerably in the depolarized fibers. The depressing ability of DAAE is assumed to be due to high concentration of the drug in the membrane owing to its high solubility in lipids. The slowing of sodium reactivation and inactivation shows the existence of a receptor for DAAE linked with a h-gate. Prolonged antiarrhythmic action is accounted for by a very slow recovery of sodium current after the drug superfusion. A stronger effect of DAAE on sodium current in the depolarized fibers is likely to point to its selective action on ischemic tissue. PMID- 6293614 TI - [Effect of diethylamine analog of ethmozine on parameters of sodium current in isolated rat cardiomyocytes]. AB - Voltage clamp experiments were made on ezymically isolated and internally perfused rat cardiac cells. The effect of a diethylamine analog of ethmozine (DAAE) on sodium current (INa) was tested when the drug was applied inside or outside the cell. It was found that the effect of DAAE (8 X 10(-6) g/ml) on INa was asymmetrical: after DAAE addition outside the cell, the amplitude of INa was effectively suppressed. Thus, 5 minutes after DAAE action the maximal value of INa in a voltage-current relationship was 20% of the control value without significant changes in the kinetics of INa. When the DAAE was added inside the cell preferentially, the inactivation time constant was increased without significant changes in the amplitude of the maximal INa. The same results were obtained with pronase (1 mg/ml) added inside the cell. It was supposed that as compared to ethmozine, the DAAE possesses a supplementary binding site on the cardiac cell membrane possibly linked to the structures responsible for inactivation processes. PMID- 6293615 TI - [Effect of epinephrine on the level of free radicals in human plasma and erythrocytes]. AB - The electron-spin resonance method was applied to examine human plasma and red cells at a temperature of 77 degrees K and variation in the level of free radicals (FR) under the effect of adrenaline. In plasma, the signals of transferrin (g approximately 4.26), ceruloplasmin (g approximately 2.05), and FR (g approximately 2.0024-2.0029 and delta H 6-8 Oe) were registered, while in red cells, the signals of hemoglobin (g approximately 6.00), superoxide dismutase (g approximately 2.063), and flavosemiquinone (g approximately 2.0030-2.0040 and delta H 12-15 Oe). Addition of adrenaline entailed an increase in the level of FR and a lowering of the ceruloplasmin signal intensity in plasma. The level of FR in red cells was found to be elevated. The mechanisms of the phenomena described are discussed. PMID- 6293616 TI - [Reactivity of B lymphocytes in mouse spleen to epinephrine during immune response]. PMID- 6293617 TI - [Function of hormone-secreting cells cultured under continuous perfusion with nutrient medium]. AB - The data are presented as to prolactin and ACTH secretion by rat adenohypophysis cells and insulin secretion by pancreatic beta-cells of the newborn rats during cell cultivation on flat synthetic membranes with pores differing in diameter under continuous medium perfusion. The cultured cells were discovered to be viable and to possess secretory activity. Secretion of prolactin and ACTH and that of insulin in the appropriate cultures increases in the course of long-term incubation under continuous medium perfusion. It is concluded that application of the porous membranes enables a selective study of the release of one or another hormone into the medium. It is believed that cultivation of hormone-secreting and hormone-susceptible cells under closed- and unclosed-type nutrient medium perfusion is fairly promising. PMID- 6293618 TI - [Activity of erythrocyte membrane Na,K-ATPase in rats with experimental botulism]. AB - The effect of type C botulinum toxin on Na, K, Mg-ATPase activities of erythrocyte membranes of white rats was studied in experiments in vivo and in vitro. The activity of Na, K, Mg-ATPase was found to be markedly inhibited in the preclinical period of poisoning, 2 hours after intraperitoneal injection of the toxin. In this case Mg-ATPase activity noticeably increased. In the presence of the development of a grave paralytic syndrome one day after intraperitoneal injection of the toxin, the activity of Na, K-ATPase of the erythrocyte membrane remained decreased as was the case in the preclinical period of poisoning, whereas the activity of Mg-ATPase returned to normal. The experiments in vitro with preincubation of erythrocyte membranes with botulinum toxin in the concentrations corresponding to the mean calculated ones in the experiments in vivo demonstrated inhibition of Na, K-ATPase. The magnitude of Mg-ATPase activity remained virtually unchanged in all the modifications of the experiments with boiled and native botulinum toxin. The in-vivo experiments with intraperitoneal injection of glutathione and unithiol to the pretreated animals attested to normalization of Na, K-ATPase in the preclinical period of poisoning, with this normalization being brought about by unithiol. In the in-vitro experiments with addition of unithiol or glutathione into the incubation medium, each of the donators of sulphhydryl groups prevented Na, K-ATPase inhibition with botulinum toxin. PMID- 6293619 TI - [Cyclic nucleotides (cAMP and cGMP) in the dog plasma during extracorporeal hemocarboperfusion]. AB - The concentrations of cyclic nucleotides (cAMP and cGMP) were measured in blood plasma of dogs subjected to extracorporeal hemocarboperfusion. In spite of complete and irreversible absorption of cyclic nucleotides by the coal absorbent, SKN-2M, the concentration of cGMP in plasma remained within normal. The concentration of cAMP increased 1.5-2.5-fold. The rise in the cAMP concentration was not connected with injection of sodium thiopental and heparin before hemocarboperfusion. It is assumed that cyclic nucleotides play the role of circulating intercellular regulators in blood plasma. These regulators are necessary for the normal course of metabolic processes. PMID- 6293620 TI - [Enzymatic detoxication of superoxide anion radical and lipoperoxides in the intima and media of the aorta in atherosclerosis]. PMID- 6293621 TI - [Role of blood lipoproteins in adaptive reorganization of the rat liver mitochondria]. AB - It was shown in experiments in vitro that high density lipids (HDL), very low density lipids (VLDL) and cAMP taken separately did not affect the kinetics of mitochondrial swelling in the liver of control and 72-hr-fasting rats. ApoHDL and particularly ApoVLDL obtained by delipidation of appropriate lipoproteins had pronounced capacities to stimulate the process of swelling. Mitochondrial swelling increased to an ever greater degree under the action of cAMP coupled with apoproteins. The maximum stimulation of the swelling was attained as a result of administering apoVLDL in conjunction with cAMP. The effects of enhanced swelling were the most remarkable in experiments with mitochondria from the control animals. PMID- 6293622 TI - [Purinergic effect of ethymisole]. AB - A study was made of the mechanism of action of ethimizole, a drug synthesized as central stimulant in terms of structural similarity to caffeine but possessing some antagonistic pharmacological effects as regards caffeine action. Experiments on rats given intraperitoneal injections of ethimizole demonstrated a considerable increase in adenylate cyclase activity of the brain and in the ATP level. Caffeine was found to prevent the stimulant action of ethimizole on energy metabolism and adenylate cyclase activity, while adenosine to potentiate this action. The possibility of ethimizole action on brain adenosine receptors is analyzed. PMID- 6293624 TI - [Effect of ethmozine on action potentials and myocardial contraction in guinea pigs]. AB - Ethmozine decreased the maximum rate of action potential rise (Vmax) in a dose dependent manner. Using the Scatchard plot the apparent dissociation constant was calculated to be 1.52 X 10(-5) g/ml. Ethmozine also decreased the force of contraction in the concentration range between 1 X 10(-6) and 1 X 10(-4) g/ml with the apparent dissociation constant obtained from the Scatchard plot being equal to 1.48 X 10(-5) g/ml. The linear correlation coefficient between the decrease in Vmax and the decrease in the force of contraction was found to be equal to 0.998. Negative inotropic action of ethmozine was less pronounced when the stimulation frequency had been switched from 0.8 to 0.1 Hz. The decrease in Vmax under the action of ethmozine (3 X 10(-5) g/ml) was diminished from 56 +/- 7% (0.8 Hz) to only 3 +/- 8% (0.1 Hz). This was accompanied by the decrease in the negative inotropic effect: from 58 +/- 9% (0.8 Hz) to 16 +/- 15% (0.1 Hz). It was assumed that the negative inotropic action of ethmozine was mediated by the Na--Ca exchange, which was inhibited by the decrease of the intracellular Na+ concentration due to the blockade of sodium channels by ethmozine. PMID- 6293623 TI - [Mechanism of action of cyproheptadine (peritol) on the hypothalamo-pituitary adrenal axis]. AB - The present paper is concerned with the mechanism of action of the blocker of serotonin receptors, cyproheptadine (peritol) on the hypothalamohypophyseal adrenal system activity. It has been established in male rats that the inhibitory, antiserotonin action of peritol on the indicated system activity depends on the rate and rhythmicity of the drug administration, and, to a less degree, on its dosage. Peritol exerts a specific antiserotonin action via the CNS and by acting on some regulatory mechanism in peripheral tissues (adenylate cyclase system, corticosterone secretion). PMID- 6293625 TI - The red cell membrane skeleton: recent progress. PMID- 6293626 TI - Platelet aggregation and exposure of fibrinogen receptors by prostaglandin endoperoxide analogues. PMID- 6293627 TI - Suppression of estrogens with aminoglutethimide and hydrocortisone (medical adrenalectomy) as treatment of advanced breast carcinoma: a review. AB - Fifty to sixty percent of postmenopausal women with estrogen receptor positive metastatic breast cancer respond objectively to surgical ablation of the pituitary or adrenal glands. Several investigators have recently developed medical alternatives to surgical ablative therapy for these patients. This review describes one of these strategies, the inhibition of estrogen synthesis with the enzyme inhibitor aminoglutethimide (AG). Aminoglutethimide blocks several cytochrome P-450-mediated steroid hydroxylation steps including those required for cholesterol to pregnenolone conversion and for the aromatization of androgens to estrogens. In women with metastatic carcinoma, a regimen including 1,000 mg of AG and 40 mg of hydrocortisone as replacement glucocorticoid was administered daily. Clinical studies revealed a 32% objective response rate to AG-HC in unselected patients, and a 52% response in women with estrogen receptor positive tumors. Randomized trials revealed that AG-HC produced objective regression as frequently as surgical adrenalectomy (Ag-HC + 53% vs. surgical adrenalectomy (43%, p = NS), and as surgical hypophysectomy (AG-HC 47% vs. hypox 21%, p + NS). Comparison of AG-HC administration with antiestrogen treatment suggested an equal rate of response to either therapy. Preliminary data document responses to AG in antiestrogen-resistant patients. Current studies do not allow precise recommendations regarding the sequence of use of antiestrogens and AG-HC. PMID- 6293628 TI - Prolactin binding by human mammary carcinoma: relationship to estrogen receptor protein concentration and patient age. AB - Biopsy specimens of 55 human mammary carcinomas (38 primary and 17 metastatic) were assayed for prolactin receptors (PrlR). Prolactin bound specifically to 32 (58%) of the tumor biopsy specimens. The apparent Kd for PrlR in individual tumors ranged from 15 pM to 2.3 nM (mean 600 pM, n = 5) and the concentration of PrlR ranged from 0 to 44.5 fmoles/mg protein. Estrogen receptors (ERP) were also detected in 28 of the 32 tumors which had PrlR. Overall, there was no correlation between PrlR and ERP. However, the mean concentration of PrlR was significantly higher (p less than 0.01) in tumors with 6-100 fmoles/mg protein ERP (approximately 13 fmoles PrlR) than in tumors with either less than 6 or greater than 250 fmoles ERP (4.0 +/- 0.4 and 6.5 +/- 1.8 respectively fmoles PrlR). Analysis of PrlR concentration as a function of patient age also showed no overall correlation, but the mean PrlR in tumors from women aged 60-70 was significantly higher (p less than 0.01) than in those from either younger or older women. A higher concentration of PrlR was observed in tumors which were classified histologically as medium or well differentiated (6.1 +/- 1.2 and 11.1 +/- 2.1, respectively) than in those classified as poorly differentiated (3.3 +/- 1.2) (p less than 0.03). There was a negative correlation between PrlR concentration and membrane yield from the tumors (r = 0.43, p less than 0.02). The membrane yield correlated with the ratio of tumor cells to stroma (histologically) (r = 0.63, p less than 0.001). In tumors from 12 patients with metastatic disease on whom follow up after endocrine-related therapy was available, the mean PrlR concentration was significantly higher in the non responding group (8.2 +/- 3.0) than in the responding group (3.4 +/- 4.2, p = 0.05). PMID- 6293629 TI - Total prolactin binding sites in human breast cancer biopsies. PMID- 6293631 TI - Transmission and immunity of bovine papilloma in Iraq. PMID- 6293630 TI - A phase II trial of tamoxifen, premarin, methotrexate and 5-fluorouracil in metastatic breast cancer. AB - Complete remissions in patients with metastatic breast cancer using endocrine therapy or chemotherapy are infrequent. Breast tumors are known to be heterogeneous with respect to estrogen receptor status, and the low complete remission rate may be related to this biochemical heterogeneity. Based on laboratory experiments using human breast cancer cells in tissue culture, a phase II protocol was designed using tamoxifen, premarin, methotrexate, and 5 fluorouracil. Thus far, twenty-nine (29) patients have been entered into this study and twenty-five (25) are currently evaluable for response. Overall response rate was 72%, and 14 of 25 (56%) attained a complete remission. Toxicity was minimal. Median nadir white blood cell count was 5,800 and median nadir platelet count was 252,000. In summary, this combination chemo-hormonal therapy regimen is effective with a more than 50% complete remission rate and minimal toxicity. PMID- 6293632 TI - Carry-over of dinitramine, triallate, and trifluralin to the following spring in soils treated at different times during the fall. PMID- 6293633 TI - [Localization of Na+, K+-ATPase in the rat submandibular gland using an autoradiographic technique]. PMID- 6293635 TI - Determination of dependence of binding parameters on receptor occupancy. PMID- 6293634 TI - A kinetics model of abrin binding in a virus transformed lymphocyte cell culture. PMID- 6293636 TI - Viruses and squamous neoplasia of the lower female genital tract: an update. PMID- 6293637 TI - The effect of desipramine upon central adrenergic function in depressed patients. AB - Eleven drug free patients meeting Research Diagnostic Criteria for Major Depressive Disorder have been treated with desipramine and given a clonidine infusion after 0, 1 and 3 weeks of treatment. The sedative and hypotensive effects of clonidine were significantly inhibited after three weeks of treatment with desipramine: a similar interaction was seen after one week of treatment although this just failed to reach statistical significance. The growth hormone (GH) response to clonidine was initially impaired, but increased significantly after one week of treatment. A significant reduction in the GH response occurred during the second and third weeks of treatment with desipramine. This last finding is interpreted as evidence of adaptive change of alpha 2 adrenoceptors: the other changes can be explained by the known ability of desipramine to block the re-uptake of noradrenaline. PMID- 6293638 TI - Converting-enzyme inhibitor enalapril (MK421) in treatment of hypertension with renal artery stenosis. AB - Enalapril maleate (MK421), a new inhibitor of angiotensin converting enzyme, in single daily doses of 1.25-40 mg was assessed in five patients with hypertension and renal artery stenosis. Only small falls in plasma angiotensin II concentrations were seen at doses less than 10 mg; even with 10 and 20 mg, angiotensin II concentrations had risen again 24 hours from the last dose. During long-term treatment with 10-40 mg daily all patients achieved good blood-pressure control. No significant changes of body sodium or potassium values were seen. The drug was well tolerated with no serious side effects. These findings are evidence of the efficacy and acceptability of enalapril in the medical management of hypertension with renal artery stenosis. PMID- 6293639 TI - Pathogenic microbial flora of genital ulcers in Sheffield with particular reference to herpes simplex virus and Haemophilus ducreyi. AB - The pathogenic microbial flora of genital ulcers in 161 (80 men and 81 women) unselected patients was studied prospectively. In only one case was Treponema pallidum responsible whereas herpes simplex virus was considered to be the cause of 130 (80.8%) genital ulcers. H ducreyi was isolated from 46 (28.6%) patients, most commonly as a secondary pathogen in herpetic lesions. Two or more pathogens were isolated from the ulcers in 67 (41.6%) patients, and in 21 (13%) patients no pathogens were isolated. Our results indicate an urgent need for antiviral treatment to reduce the local reservoir of genital herpes, challenge traditional concepts about the prevalence of H ducreyi in Britain, and call for a reappraisal of its role in the causation of genital ulcers. PMID- 6293640 TI - Preparation and immunogenicity of vaccine Ac NFU1 (S-) MRC towards the prevention of herpes genitalis. AB - A subunit antigenoid vaccine, Ac NFU1 (S-) MRC, was used to prevent primary herpes genitalis in 60 subjects considered to be at risk of this infection. There was no evidence of serious local or general side effects. Neutralising antibody responses were detected in 59% and 90% of subjects receiving the low and high doses of vaccine respectively; immunoprecipitating antibody was detected at a lower frequency, namely in 23% and 43% of subjects receiving the low and high doses respectively. After a mean follow-up period of 18 months none of the vaccinated subjects contracted herpes genitalis after completing the vaccination course. PMID- 6293641 TI - Choline and PAH transport across blood-CSF barriers: the effect of lithium. AB - The components of the blood-CSF barrier responsible for the transport of p aminohippuric acid (PAH) and choline from CSF to blood were identified using in vitro preparations of frog choroid plexus and arachnoid membranes. Choline was transported out of CSF across the arachnoid, while PAH was transported out across the choroid plexus. Probenecid and ouabain blocked both processes. The effect of Li on these transport processes was tested by the addition of 5 mM LiCl to the incubation media. Li increased, by a factor of two, choline transport across the arachnoid, but there was no effect of Li on PAH transport across the plexus. Lithium was passively transported across the choroid plexus, and we suggest that the major transport pathway is through the tight junctions. The steady-state distribution of Li between the choroidal epithelium and the incubation medium was only half that expected for passive distribution. This suggests the existence of sodium/lithium countertransport in these epithelial cell membranes. PMID- 6293642 TI - Independent in vitro regulation by the D-2 dopamine receptor of dopamine stimulated efflux of cyclic AMP and K+-stimulated release of acetylcholine from rat neostriatum. AB - Two types of dopamine receptors whose stimulation affect cAMP efflux (and by inference formation) could be identified in rat neostriatum. One type of receptor, called D-1 receptor, increased cAMP efflux whereas stimulation of a second type of dopamine receptor, called D-2 receptor, was followed by a reduction in cAMP efflux induced by stimulation with a D-1 receptor agonist. D-2 receptor agonists inhibited the effects of D-1 receptor agonists on cAMP efflux in a non-competitive way. These inhibiting effects of D-2 receptor agonists occurred also in the absence of Ca2+-ions which could imply that some of the D-2 receptors are located on cells possessing D-1 receptors. The dopamine receptor mediating inhibition of the release of radiolabeled acetylcholine (ACh) in the neostriatum appeared to have the same pharmacological characteristics as the D-2 dopamine receptor mediating the inhibition of the D-1 receptor agonist induced cAMP efflux. Selective D-2 receptor agonists like LY 141865 and RU 24926 stimulated this receptor while the D-1 receptor agonist SKF 38393 was inactive. Effects of the selective D-2 receptor agonists could be antagonized by (-) sulpiride, a selective D-2 receptor antagonist. Although the pharmacological characteristics of the dopamine receptors mediating inhibition of both ACh release and (D-1 dopamine receptor agonist induced) cAMP efflux appeared to be similar, drugs stimulating cAMP efflux did not affect ACh release or LY 141865 induced inhibition of ACh release from rat neostriatum. Therefore it is still questionable whether the dopamine receptor mediating inhibition of both ACh release and cAMP efflux is one and the same functional entity. PMID- 6293643 TI - Modification of transmission in the cuneate nucleus by raphe and periaqueductal gray stimulation. AB - In anesthetized decerebellate cats, with additional decerebration or decortication and with one of two types of cervical spinal cuts which either eliminated the dorsal half of the spinal cord or spared the dorsal funiculi, conditioning stimulation in the raphe nuclei or periaqueductal gray modified transmission in the cuneate nucleus. This was shown by 4 types of recordings: surface potentials, somatic afferent excitability testing, lemniscal tract response and extracellularly recorded single neuron activity. PMID- 6293644 TI - Effects of calcitonin on rat extrapyramidal motor system: behavioral and biochemical data. AB - The effects of i.v.c. injection of human and salmon calcitonin on biochemical and behavioral parameters related to the extrapyramidal motor system, were investigated in male rats. Calcitonin injection resulted in a potentiation of haloperidol-induced catalepsy and a partial prevention of apomorphine-induced hyperactivity. Moreover calcitonin induced a significant decrease in nigral GAD activity but no change in striatal DA and DOPAC concentration or GAD activity. The results are discussed in view of a primary action of calcitonin on the striatonigral GABAergic pathway mediating the DA-related behavioral messages of striatal origin. PMID- 6293645 TI - Evidence that cholecystokinin is a neurotransmitter of olfaction in nucleus olfactorius anterior. PMID- 6293646 TI - Biogenic antagonists of the nicotinic receptor: their interactions with erabutoxin. AB - The hypothesis that the sensitivity of the nicotinic ACh-receptor is reduced by some neurotransmitters was evaluated by studying the interaction between these neurotransmitters and erabutoxin-b (ETX-b), known to bind irreversibly with the specific ACh-receptor site. It was found that the blocking action of ETX-b was apparently prevented by previous application of 5-HT, whereas it was not prevented by application of catecholamine (CA). These results indicate that 5-HT blocks the nicotinic ACh-receptor by interacting with the specific ACh binding site, whereas CA blocks it by interacting with an allosteric site of the ACh receptor ionic channel complex. PMID- 6293647 TI - Excitatory effects of ACTH on noradrenergic neurons of the locus coeruleus in the rat. AB - Responses of presumed noradrenaline-containing neurons in locus coeruleus (LC) to iontophoretically applied ACTH were determined. The full sequence ACTH1-39 and the partial sequence ACTH1-24 were excitatory on the majority of neurons tested. The partial sequence ACTH27-39 was without effect on most neurons. The presence of ACTH-containing fibers in LC together with the present results may suggest a controlling influence of the peptide on the noradrenaline system. PMID- 6293648 TI - Ischemic pain nonsegmentally produces a predominant reduction of pain and thermal sensitivity in man: a selective role for endogenous opioids. AB - Ischemic pain was produced by a blood pressure cuff placed to the arm of healthy human subjects for 15 min which produced a mean pain score of 59% (visual analogue scale). Ischemia induced a significant dental pain threshold elevation (mean 67%) and 2 mg of naloxone did not reduce it. Thermal sensitivity of the upper lip had a tendency to reduction during ischemia and 2 mg of naloxone reduced this effect. Tactile thresholds in the forehead or in the contralateral arm were not markedly elevated. Neither ACTH nor prolactin level in the plasma was related to the dental pain threshold elevation during ischemia. The findings of the present study suggest that ischemic pain nonsegmentally produces a predominant inhibition of responses to thin afferents. Endogenous opioids may markedly contribute to the reduction of thermal sensitivity induced by ischemia, but their contribution to dental pain threshold elevations seems to be less important. Stress or other adenohypophyseal mechanisms involving the release of ACTH or prolactin do not explain the effects of ischemia found in the present study. PMID- 6293649 TI - Sleep and medial reticular unit responses to protein synthesis inhibitors: effects of chloramphenicol and thiamphenicol. AB - This study utilized a newly developed combination push-pull cannula/microdrive microwire device to record single-unit activity within the diffusion field of substances introduced into the brainstem. Single unit activity within the midbrain and pontine medial reticular formation (RF) and sleep were recorded following either perfusion or oral administration of protein synthesis inhibitors, chloramphenicol and thiamphenicol. Chloramphenicol administration led to significant reductions in the frequency, but not the duration, of individual REM episodes without altering slow-wave sleep. During control experiments in slow wave sleep, incipient REM was characterized by substantial increases in medial RF unit activity which occurred in association with the appearance of PGO spikes. On the other hand, discharge rates of medial RF neurons were markedly attenuated by chloramphenicol but were not consistently affected by thiamphenicol. These findings suggest that reduced unit activity contributes to the REM-suppressive effects of protein synthesis inhibitors. PMID- 6293650 TI - An electron microscopic study on enkephalin-like immunoreactive nerve fibers in the celiac ganglion of guinea pigs. AB - Enkephalin-like immunoreactive nerve fibers in the celiac ganglion of guinea pigs were characterized by a high population of large granular vesicles mixed with small clear vesicles. The immunoreactive material is confined to the large granular vesicles. The immunoreactive nerve fibers formed many axo-dendritic as well as axo-somatic synapses and also formed a few synapses with presumed preganglionic axons containing numerous vesicles. The immunoreactive fibers were regarded as presynaptic at these synapses. These findings suggest that enkephalin might play a role as a neurotransmitter or neuromodulator in the ganglionic transmission of this prevertebral ganglion. PMID- 6293651 TI - Responses of vestibular-nerve afferents in the squirrel monkey to externally applied galvanic currents. PMID- 6293652 TI - Freeze-fracture study of membranous cytoplasmic bodies of cortical neurons in feline GM1-ganglioside storage disease. PMID- 6293653 TI - Localization of [3H]2-deoxyglucose at the cellular level using freeze-dried tissue and dry-looped emulsion. PMID- 6293654 TI - Nicotinic and muscarinic reactive sites in mammalian glomus cells. AB - Nicotinic and muscarinic sites on glomus cell membranes of cats, rabbits and mice were determined in carotid body slices. Cells were impaled under Nomarski optics. Resting potentials were 11.1-73.1 mV and input resistances were 11-250 M omega. Nicotine, pilocarpine or bethanechol depolarized glomus cells and their input resistance decreased. Curare or alpha-bungarotoxin reduced nicotine effects. Atropine had similar effects on the responses to pilocarpine or bethanechol. PMID- 6293655 TI - alpha 2-Adrenergic receptors inhibit catecholamine secretion from bovine adrenal medulla. AB - Catecholamine secretion evoked by carbamylcholine from isolated bovine adrenal medullary cells was inhibited by alpha 2-agonist, clonidine, in a dose-dependent manner with IC50 value of 2.8 X 10(-5) M. [3H]Clonidine bound to adrenal medullary membranes with high affinity and saturable characteristics. These results suggest that alpha 2-adrenergic receptors which exist on adrenal medullary cells have inhibitory effect on the secretion of catecholamine from the cells. PMID- 6293656 TI - Effects of anions on calcium component in sensory nerve terminal of frog muscle spindles. AB - An increase in the Ca2+ component following individual sodium spike in spontaneous discharges was observed in the isolated sensory terminal of the frog muscle spindle perfused with isotonic sodium solution with a group of anions, of which the size in the aqueous solution ranged from 0.74 to 1.32 times that of hydrated sodium ion. The effects of these anions was counteracted with divalent cations. It is hypothesized that anions similar in size to hydrated sodium ions may form an anion-cation complex at the interaction site of the Na+ carrier, whereby the Na+ may enter the Ca2+ channel. This may be inactivated by the divalent cations. PMID- 6293657 TI - Is a retrosplenial (cingulate) pathway involved in the mediation of high frequency hippocampal rhythmical slow activity (theta)? AB - In previous experiments we demonstrated that in rats there are two kinds of hippocampal rhythmical slow activity patterns (RSA or theta) as defined by the dominating EEG frequencies. RSA with a frequency of 6-8 Hz appeared during exploratory behavior (locomotion), whereas stimulation of the dorsomedial hypothalamus (DMH) elicited RSA with frequencies of 8-12 Hz. To determine the neural pathways involved in the mediation of these two types of RSA, local injections of tetracaine were made either in the medial septum or in the cingulate cortex in order to reversibly interrupt the functional activity of these loci. Blockade of the medial septum suppressed the 6-8 Hz 'walking associated' RSA in the hippocampal EEG, but had no effect on the 8-12 Hz DMH driven RSA. On the other hand, a tetracaine injection into the cingulate cortex selectively blocked the high-frequency RSA elicited by DMH stimulation, but had no effect on the 6-8 Hz 'walking-associated' RSA. Both effects disappeared between 30 and 90 min after tetracaine injection. We conclude that the DMH-driven RSA is mediated by the cingulum and/or fibers traveling through the cingulate cortex (retrosplenial region) and thus, that this type of RSA operates without septal involvement. PMID- 6293658 TI - GK(Ca)-dependent cyclic potential changes in the sensory nerve terminal of frog muscle spindle. AB - Spontaneous cyclic hyperpolarizations along the sensory nerve terminal of frog muscle spindles were observed during the application of 1-9 nA depolarizing currents across an air-gap on which the axon was bridged. An increase in the current intensity increased the amplitude and duration of the cyclic changes. Upon subthreshold depolarization, single or repetitive hyperpolarizations could be elicited after a brief electric pulse or during stretch of the receptors, respectively. The threshold was decreased in higher Ca2+, Sr2+ or Ba2+ solutions. The cyclic changes were reversibly blocked by K+- or Ca2+-blockers and quinine. These results suggest that the changes are due to GK(Ca). The site of origin of the changes was at the branching node in the capsule, as confirmed by the following results: (1) the cyclic changes were abolished upon inactivating the node by UV-irradiation; (2) in normal Ringer's solution, the rate of afferent impulses, which reflects the membrane potential at the encoding site along the non-myelinated filaments, was unmodified by the cyclic changes and was independent of the intensity of the polarizing currents within a certain range; however, it was sensitively dependent on this intensity after treatment with K+ blockers; (3) the amplitude of the impulses reaching the branching node was attenuated during the cyclic changes, but not after GK-blockade. PMID- 6293659 TI - Increased sensitivity of neurons to angiotensin II in SHR as compared to WKY rats. AB - Angiotensin II (ANG II)-sensitive septal neurons in the brain of stroke-prone spontaneously hypertensive rats (SHR-sp) and of normotensive Wistar-Kyoto rats (WKY) were investigated for possible differences at receptor sites. ANG II, the competitive ANG II-antagonist saralasin, and acetylcholine (ACh), were applied microiontophoretically onto neurons of the lateral septal area. ANG II-evoked neuronal firing which was specifically inhibited by saralasin occurred at a significant lower threshold in SHR-sp (23%) and showed an extended postactivity (340%) as compared to the age-matched WKY controls. In contrast, the activity due to ACh remained similar in both strains. PMID- 6293660 TI - The distribution of [3H]kainic acid binding sites in rat CNS as determined by autoradiography. AB - The distribution of [3H]kainic acid (KA) binding sites in the rat CNS was determined by in vitro autoradiography. KA sites are distributed throughout the CNS gray matter in an anatomically specific pattern with telencephalic structures and the cerebellum accounting for the majority of the binding. These results, together with our previous finding that KA sites are greatly enriched at the synapse, suggest that KA binding sites are associated with select terminal fields, and hence may be involved in neurotransmission in certain CNS pathways. PMID- 6293661 TI - Fluorescence histochemical methods for the study of peptide hormone-producing cells. AB - Fluorescence histochemical methods for the demonstration of specific residues in peptides and proteins are reviewed: Formaldehyde-ozone for NH2-terminal tryptophan, formaldehyde-HCl for tryptophan regardless of position in the peptide, OPT for NH2-terminal histidine, formaldehyde-fluorescamine for "protected" amino groups, nitroso-naphthol for tyrosine, and phenanthrenequinone for arginine residues. The methods are potent in demonstrating granule-stored material in peptide hormone-producing cells. Also quinacrine, the fluorescent anti-malaria agent, binds to granular components, as yet unidentified, in several endocrine cell types. In many cases the fluorescence histochemical methods seem to demonstrate peptides and proteins distinct from the known hormones. PMID- 6293662 TI - Spinal cord grafts: an intraocular approach to enigmas of nerve growth regulation. AB - The possible usefulness of intraocular transplantation in studies of spinal cord growth and regeneration has been evaluated. Defined segments of fetal rat spinal cord were grafted to the anterior chamber of the eye of adult rats. Such grafts become vascularized from the host iris, grow and develop neuron types, myelinated fiber bundles, astroglial populations (as shown by GFA-immunoreactivity), and electrical activity reminiscent of such features in normal spinal cord tissue. The intraocular technique permits studies of intrinsic circuitries as well as conditions for formation of afferent and efferent connections with the host iris and with other central or peripheral tissues which can be grafted into contact with the spinal cord grafts. One example of an intrinsic system preserved in the grafts is a rich network of nerve fibers with enkephalin-like immunoreactivity. When combined with cerebral cortex, the enkephalin-positive neurons of the spinal cord graft are able to form only very limited projections to the cortex graft. Special emphasis was given the possible formation of adrenergic afferents to spinal cord grafts. No appreciable ingrowth of peripheral sympathetic nerves occurred. Locus coeruleus grafts have many organotypical electrophysiological characteristics and were able to innervate adjacent spinal cord grafts provided that the sensory innervation of the host iris was removed. Experiments such as these suggest that "negative neurotropic" factors may be present in spinal cord and possibly relate to the unique relationship between spinal ganglia and spinal cord. PMID- 6293664 TI - Contraction of the ovarian follicle induced by local stimulation of its sympathetic nerves. AB - Strips of bovine ovarian follicle wall, known to contain smooth muscle cells innervated by adrenergic nerves, were dissected out and exposed to transmural electrical stimulation in an organ bath. A frequency-dependent contractile response was obtained with a maximum at 8-16 Hz. The response was abolished in the presence of tetrodotoxin and inhibited by bretylium, phentolamine and reserpine. Thus, stimulation of the sympathetic nerves in the ovarian follicle releases sufficient amounts of norepinephrine to produce a contraction of its wall, an effect mediated by alpha-adrenergic receptors. PMID- 6293663 TI - Catecholamine fiber regeneration across a collagen bioimplant after spinal cord transection. AB - A cell-free bovine derived collagen matrix was used to study potential axonal regeneration in transected rat spinal cord. Rats were initially subjected to a 200 g/cm force acceleration injury at T10 and 10 days later, the spinal cord was totally transected at the injury site. Controls had their spinal cord stumps juxtaposed end-to-end following transection. Experimental rats had 3-4 mm of spinal cord tissue trimmed from the proximo-distal stumps. The semi-fluid collagen material was implanted to bridge the proximo-distal ends and after several hours, the collagen graft polymerized to a firm gel. Rats were observed for 90 days. After 90 days, animals were evaluated using somatosensory evoked potentials, local spinal cord blood flow, and catecholamine histofluorescence in and around the site of transection. Results suggest that the collagen bioimplant can support the development of anastomotic blood vessels with the cord as well as provide a non-hostile environment to regenerating spinal cord axons. PMID- 6293665 TI - Sympathetic influence on sodium-potassium activated adenosine triphosphatase activity of rabbit and rat choroid plexus. AB - Ouabain-sensitive Na+-K+-ATPase was measured spectrophotometrically in the lateral choroid plexuses of rabbit and rat. In the rabbit, a significant increase in the enzyme activity was seen at one week after unilateral sympathectomy (removal of the superior cervical ganglion), but not at three days or two weeks postoperatively, as compared with the intact, contralateral plexus. Unilateral sympathetic denervation of the rat's choroid plexus induced a nearly 40% decrease in Na+-K+-ATPase activity at 6 days after the operation, while no effect was seen after 12 days. The results agree with a local sympathetic inhibition of CSF production in rabbit (corresponding studies on rat have not been performed), and favor the assumption that the adrenergic nerves in the choroid plexus mediate direct effects on transport functions in the plexus epithelium. PMID- 6293666 TI - [Use of silica-based products at a maximum amount of 1% in caseinates, serum proteins and milk powders to facilitate the fluidization of these powders]. PMID- 6293667 TI - [Parameters influencing poliovirus inactivation by chlorine]. PMID- 6293668 TI - [Purine metabolic defects in primary immunodeficiency]. PMID- 6293669 TI - [Experimental studies on the infection of an adult tree shrew (Tupaia belangeri yunalis) with human rotavirus]. PMID- 6293670 TI - Optimal conditions for Ca-acidic phospholipid-PO4 formation. AB - Calcium-acidic phospholipid-phosphate complexes (Ca-PL-PO4) cause hydroxyapatite (HA) deposition in vitro. The acidic phospholipids--phosphatidylserine (PS), phosphatidylinositol (PI), phosphatidylglycerol (PG), and diphosphatidylglycerol (DPG)--all form Ca-PL-PO4 complexes: Nonacidic phospholipids--sphingomyelin (SPL), phosphatidylcholine (PC) and phosphatidylethanolamine (PE)--do not form Ca PL-PO4 and do not induce hydroxyapatite (HA) deposition in low ionic strength metastable calcium phosphate solution or in synthetic lymph. The extent to which each of the acidic phospholipids reacts with calcium and inorganic phosphate, and the chemical composition of the Ca-PL-PO4 complexes is a function of pH, solution composition, the nature of the phospholipid, and the method of isolation. The addition of inorganic phosphate prior to or in combination with calcium appears to be an absolute requirement for Ca-PL-PO4 formation. PMID- 6293671 TI - Conversion of amorphous tricalcium phosphate into apatitic tricalcium phosphate. AB - Precipitated calcium orthophosphate, rapidly prepared in highly supersaturated solutions at pH 9-11, is an amorphous tricalcium orthophosphate (atomic ratio Ca/P = 3/2) of formula Ca9(PO4)6, nH2O. Kept wet at room temperature, this phosphate is hydrolyzed according to the reaction PO4(3-) + H2O leads to HPO4(2-) + OH-; a tricalcium orthophosphate series is then formed, its general formula is Ca9 (HPO4)x (PO4)6-x (OH)x, O less than or equal to x less than or equal to 1. Moreover, the amorphous phosphate is converted into apatitic phosphate at half hydrolysis (x approximately equal to 0.5). The hydrolysis occurs simultaneously with an endothermic effect [delta H = 12.0 +/- 2.0 kJ/mol Ca9(PO4)6], and the conversion with an exothermic effect [delta H = -5.9 +/- 1.0 kJ/mol Ca9(PO4)6]. PMID- 6293672 TI - Effect of fluoride on crystal growth of calcium apatites in the presence of a salivary inhibitor. AB - The effect of fluoride on the kinetics of crystal growth of calcium apatites was studied using seed crystals of hydroxyapatite coated with PRP-3, a proline-rich phosphoprotein salivary inhibitor of crystal growth. Initial precipitation rates in the presence of PRP-3, under conditions of partial inhibition, were enhanced by fluoride, effectively counteracting the inhibitory activity of the macromolecule. This rate enhancement is related to an increase in the precipitation driving force, i.e., the degree of supersaturation with respect to the precipitating phase; in this case a fluoridated hydroxyapatite. The growth of this phase takes place at the uncovered crystal growth sites which have been previously shown to be the same sites as the adsorption sites for the protein inhibitors. Explanations based on fluoride activation of secondary crystal growth sites and on the displacement of adsorbed inhibitor by fluoride are not substantiated by the present results. It has been demonstrated that under conditions of maximum coverage of available crystal growth sites by PRP-3, resulting in no apparent crystal growth, measurable crystal growth is observed upon the addition of fluoride (1 ppm). This phenomenon is best explained by the fact that at maximum PRP-3 coverage, a small number (16%) of crystal growth sites remain uncovered, which support an unmeasurable rate of crystal growth. Upon the addition of fluoride, this rate is significantly enhanced. It is suggested that the fraction of uncovered growth sites is related to steric interactions of the PRP-3 molecule in the adsorbed state. Overall, the results presented suggest that fluoride can accelerate crystal growth in an environment such as the enamel surface where the acquired pellicle is formed. PMID- 6293673 TI - ESR on the hydroxyl ion vacancies in the apatites. AB - Heating biological apatites and synthetic apatites in the stream of inert gas saturated with heavy water vapor, introduces OD ions which substitute for the hydroxyl (OH) ions and partially for the OH vacancies, depending on the heating time and the gas flow rate. This has been verified by ESR and partially by IR. Deuteration gives no serious alterations of the a-axis dimension or of the PO4 bands in the IR spectra. These findings have been interpreted as suggesting that the OH ions diffuse out and the OD ions diffuse in through specific pathways, and the diffusing ions interact permanently with none of the other ions in the apatite structure. Heating in dry inert gas results in the formation of hydroxyl ion vacancies. The ESR spectra of these samples in the g = 2.06 region have been interpreted in terms of O-2 and O-. The shortening of the a-axis dimension in this case has been accounted for based on the loss of structurally incorporated H2O and the loss of the OH ions on the hexad axis. PMID- 6293674 TI - Phosphoprotein inhibition of hydroxyapatite dissolution. AB - A chromatography column containing hydroxyapatite beads was used to study the effect of different proteins on the rate of hydroxyapatite dissolution. The four phosphoproteins tested (phosvitin, alpha sl-casein, beta-casein and kappa-casein) markedly reduced the rate of hydroxyapatite dissolution. Three nonphosphorylated proteins had a relatively smaller effect. The effect of the protein in reducing the hydroxyapatite dissolution rate has been attributed to protein binding to the surface of hydroxyapatite. The reduction in dissolution rate, expressed as the change in nmol calcium released per min per nmol of phosphoprotein bound to hydroxyapatite, increased with increasing number of phosphoserine residues of the protein. The results are consistent with the proposition that phosphoproteins have a regulatory role in mineralization processes and could provide a mechanism by which dietary and salivary phosphoproteins exert an anticariogenic effect. PMID- 6293675 TI - Orientation of adsorbed alizarin red S on hydroxyapatite. AB - Adsorption isotherms of alizarin red S on hydroxyapatite were supplemented with additional data to provide a simple model of adsorption. The model treats the adsorbing surface as a mosaic of unit cell a-c faces which have an area of 65 A2 each. The adsorbate of the model has molecular dimensions consistent with a 1:1 relationship between adsorbate and adsorption site. By comparing slurries of hydroxyapatite which differed by a factor of 4 in surface area of the solid, it was found that the isotherm based on edge-on adsorption was best simulated by the model. A method for compensating for differing degrees of surface coverage is described. PMID- 6293676 TI - In vitro formation of crystalline apatite by matrix vesicles isolated from rachitic rat epiphyseal cartilage. PMID- 6293677 TI - Preparation, analysis, and characterization of carbonated apatites. AB - A range of synthetic carbonated apatites were characterized using infrared and Raman spectroscopy, X-ray diffraction, and chemical analysis and compared to stoichiometric hydroxyapatite and dental enamel. Synthetic apatites were prepared by aqueous precipitation and by solid-state reaction at high temperatures. A method is described for the carbonate determination of apatite samples using gas chromatography. This study demonstrates that carbonate exists in two crystallographically distinct sites in the apatite crystal structure and is consistent with a carbonate-for-phosphate substitution mechanism, where four carbonate groups replace three phosphate groups. Sodium and hydroxyl ions are shown to be involved with the substitution of carbonate ions in the apatite structure. Observed sodium/carbonate and calcium/phosphorus molar ratios confirm this mechanism. Substitution of this type may explain the predominant mode of carbonate substitution in biological apatites. PMID- 6293678 TI - In vitro evaluation of editempa on hydroxyapatite formation and its effects on dental enamel. AB - N,N,N',N' ethylene diamine tetra (methylene phosphonic acid)-Editempa inhibited the formation of hydroxyapatite (HA) in vitro at 4 ppm. In the seeded crystal growth of HA at 37 degrees C, it completely inhibited the crystal growth at 0.5 ppm. C14-Editempa adsorbed to HA crystal at 37 degrees C. The maximum adsorption was 1.29 mumol/m2 of HA and the adsorption was monolayer. Compared to the natural inhibitors of HA in saliva (Statherin and Proline-rich proteins), Editempa was more effective in vitro on weight basis. At 0.3 ppm, it inhibited growth rate of the crystals by 80% while 4 to 5 ppm of the natural inhibitors were needed to get the same effect. The inhibition of rates with Editempa was not directly proportional to the area covered by it on the HA seeds (occupied less than 10% of the surface). The natural inhibitors, on the other hand, seemed to cover all the available surface. These data indicate that the inhibition with Editempa was due to binding to specific sites on the surface of the seeds. The solutions up to 5% of Editempa did not damage or etch the surface of human dental enamel in vitro at pHs 5.0 and 7.5, as evaluated by scanning electron microscopy and calcium released in the exposed solutions. PMID- 6293679 TI - Electron spin resonance study of sound and carious enamel. AB - The present electron spin resonance (ESR) study aimed to investigate chemical alterations in enamel apatite associated with carious attacks. Enamel blocks were cut from sound areas and natural white spot lesions of human molar teeth and then X-irradiated at room temperature in air. Q-band ESR analysis of these samples showed the presence of two stable signals in the spectra recorded at room temperature. One of them has been identified at CO3-(3) centers, which originate from the carbonate ion substituting in the apatite lattice. The other signal is tentatively identified as O- defects in the hexad axis of apatite lattice. A comparison of the spectra between sound and carious samples demonstrated that the intensity ratios between the O- and the g parallel component of CO3-(3), R = O /CO3-(3) (g parallel), are significantly higher in the carious samples than in the sound ones. This suggests that enamel apatite in the white spot lesions may have less carbonate ions in the crystal lattice, or have a higher producibility of the O- defects in its hexad axis. The observed alterations in enamel apatite may well be accounted for by remineralization playing a major role in the formation of white spot lesions, because this acid-dissolution study using irradiated enamel blocks showed that the R values do not increase but decrease in a simple process of demineralization. PMID- 6293680 TI - ESR identification of radiation damage in synthetic apatites: a study of the 13C hyperfine coupling. AB - Although radiogenic free radicals are widely used as molecular probes in biological calcified tissues, their nature and their location remain controversial. In the present study we have synthesized and irradiated apatites in which 13CO3(2-) ions substitute hydroxyl or phosphate ions in hydroxyapatite. The 13C-couplings show that the carbon-centered radicals are CO2- --rather than CO3(3-)--the locations of which depend on the type of carbonate and the treatment of the sample. PMID- 6293681 TI - Adenoassociated virus has a unique chromatin structure. AB - The organization of intranuclear adenoassociated virus DNA (AAV) was examined following micrococcal nuclease digestion of nuclei prepared from cells coinfected with AAV type 2 (AAV-2) and adenovirus type 2 (Ad2). Blot-hybridization analysis of the DNA with AAV-2, Ad2, and cellular DNA probes revealed that AAV-2 chromatin has a unique structure, which upon nuclease digestion gives rise to a smear of oligomeric DNA fragments from 600-2200 base pairs in length with only a very faint band about 160 base pairs and no discrete multimers. This structure was similar to, but distinguishable from, Ad2 chromatin and completely unrelated to eukaryotic chromatin. PMID- 6293682 TI - Convenient biosynthetic preparation of isomeric spin-labelled radioactive phosphatidic acids. AB - A convenient method for the enzymatic preparation of sn-3-[2-3H]phosphatidic acids carrying also 5-, 12-, or 16-nitroxide stearic acids, from sn-3-[2 3H]glycerophosphate and isolated guinea pig liver microsomes, is described in detail. The procedure allows a simultaneous preparation of three spin-labelled sn 3-[2-3H]phosphatidic acids of yields 3-3.5 mumol of each compound which is greater than 99% pure in respect to the radioactivity and which contains 25 mol% of spin-labelled fatty acids. These phosphatidic acids were approximately equally distributed between the primary and the secondary hydroxyl when 12- or 16 nitroxide stearic acids were used or predominantly (75%) associated with the secondary hydroxyl of sn-3-[2-3H]phosphatidic acid when 5-nitroxide stearic acid was present in the incubation mixture. PMID- 6293683 TI - Cytochrome c554 as a possible electron donor in the hydroxylation of ammonia and carbon monoxide in Nitrosomonas europaea. AB - Mechanism of ammonia oxidation was studied in the reconstituted system of Nitrosomonas membrane fraction plus the Nitrosomonas cytochrome c554. The cytochrome c554 was reduced by hydroxylamine, hydrazine, and ammonia and the reduced cytochrome was oxidized upon the addition of ammonia or carbon monoxide. The oxidation of carbon monoxide in the presence of hydroxylamine or hydrazine was studied as a possible assay method for ammonia hydroxylase where hydroxylamine or hydrazine was supplying the reducing power required for the hydroxylation of carbon monoxide. The stoichiometry of the reaction, Km values for substrates, and effects of pH and inhibitors were investigated. It is concluded that carbon monoxide, a competitive inhibitor for ammonia oxidation, is an alternate substrate for ammonia hydroxylase using the reduced cytochrome c554 as the reducing power. PMID- 6293684 TI - Effect of ACTH or zinc treatment on plasma aldosterone and corticosterone levels and on the in vitro steroid output from adrenocortical cells. AB - We have studied the effect of in vivo treatment with two forms of ACTH (Synacthen and Duracton) and of zinc hydroxide on plasma corticosteroid levels from adult Long Evans female rats. The corticosteroid output by isolated zona glomerulosa cells in vitro was also studied. Dose-response experiments showed that, after 2 days of treatment, Synacthen caused a 2.5- and 25.9-fold increase in plasma aldosterone and corticosterone levels, respectively, while maximal increases of 1.7- and 5.6-fold were obtained following treatment with Duracton. In contrast to elevated plasma steroid levels, the basal aldosterone and corticosterone output by isolated zona glomerulosa cells was significantly decreased for all doses of Synacthen administered. The 8 IU/day treatment with Synacthen produced an 86% diminution of aldosterone output while a treatment of 32 IU/day with Duracton gave only a 48.8% decrease. Concomitantly the Synacthen treatment provoked a high mitotic response in the zona glomerulosa cells (fivefold over control). A 2-week treatment with Synacthen resulted in elevated plasma aldosterone and corticosterone levels and produced a 92% diminution of aldosterone output by isolated zona glomerulosa cells. The aldosterone and corticosterone output from these cells was not enhanced by the addition of ACTH in the incubation media; zona glomerulosa cells of animals treated with Synacthen were no longer responsive to ACTH stimulation in vitro. The effect of a 2-day treatment with zinc hydroxide on plasma aldosterone and corticosterone levels and on steroid output by isolated adrenocortical cells was different from that of Synacthen. This meant that zinc was not the active principle of the Synacthen preparation. Our results indicate that long-term treatment with ACTH provokes profound functional changes at the adrenocortical zona glomerulosa level. PMID- 6293685 TI - Characterization of the electrophoretic mobility mutation in the N protein of the tsD1 mutant of vesicular stomatitis virus New Jersey serotype. AB - Some isolates of the temperature sensitive mutant tsD1 of complementation group D of vesicular stomatitis virus of New Jersey serotype have a nucleocapsid (N) protein which shows an increased electrophoretic mobility on sodium dodecyl sulfate--polyacrylamide gel electrophoresis (SDS-PAGE) when compared with wild type. Utilizing techniques involving specific chemical cleavage at tryptophan or methionine residues, as well as enzymatic cleavage with carboxypeptidases A and B, we have determined that residues near the carboxyterminus are responsible for the electrophoretic difference of the mutant protein. We have further shown that there are no differences in the tryptic peptides of the mutant compared with the wild type or a non-ts revertant in this region of the protein. We have identified a tryptic peptide located outside the relevant carboxyterminal region which is distinct in mutant and revertant. We conclude that the mutation producing the aberrant electrophoretic mobility of N protein of the tsD1 mutant is a missense point mutation located at least 40 amino acid residues from the carboxyterminus and which interacts with a more proximal carboxyregion so as to influence electrophoretic mobility on SDS-PAGE. PMID- 6293686 TI - Evidence for bluetongue virus in Canada: 1976-1979. AB - Since the identification of approximately 1 400 bovine serological reactors to bluetongue virus in the Okanagan Valley of British Columbia in 1976, there has been no evidence of virus establishment in Canada. No clinical signs suggestive of bluetongue were observed. It was not possible to demonstrate viral activity at the time the seropositive animals were detected and subsequent serological testing supports the hypothesis that the virus has not become endemic or indeed survived in Canadian cattle populations. This combined with the dramatic reduction in prevalence of serological reactors in the years following the initial slaughter suggests that viral activity occurred in the Okanagan Valley prior to 1976 and disappeared. There has been no evidence for transmissions different from that expected of a classical arbovirus; that is, no evidence of "vertical" transmission. PMID- 6293687 TI - Experimental fetal infection with bovine viral diarrhea virus. I. Virological and serological studies. AB - The serological and virological results of an experimental infection of bovine fetuses with bovine viral diarrhea virus are presented. Four fetuses, 120-165 days gestational age, were inoculated in utero with a second passage virus strain. Two fetuses received a sham-inoculum. A humoral immune response in the virus-inoculated fetuses, was demonstrated three weeks later. In three fetuses only IgM and IgG1 were detectable. The serum from the fourth fetus also contained IgG2 and IgA. Bovine viral diarrhea virus-neutralizing antibodies were detected in two fetuses. These two fetuses inoculated at 135-150 days gestational age, represent the youngest reported bovids, giving a specific response in three weeks following an experimental infection with bovine viral diarrhea virus. The fetal sera did not contain heat-labile factors, which could mediate the neutralization. The virus was not reisolated from any of the fetuses, but viral antigen was nevertheless demonstrated by immunocytochemical methods in sections of several of the fetal organs, primarily lymphoid tissues. PMID- 6293688 TI - Experimental fetal infection with bovine viral diarrhea virus. II. Morphological reactions and distribution of viral antigen. AB - The effect of an infection with bovine viral diarrhea virus on fetal bovine tissues as well as the tissue-localization of viral antigen are described. Four bovine fetuses, 120-165 days of gestation, were inoculated in utero with a second passage virus strain. Lymphoid tissues were studied by light and electron microscopy. The infection induced precocious development of the secondary lymphoid organs. Characteristic changes were seen in postcapillary venules, cells of the mononuclear phagocyte system and lymphoid follicles. In all four fetuses the thymus was hypoplastic. In three fetuses this implicated a morphological immaturity, but no actual pathological alterations. In the fourth fetus, the hypoplasia was caused by necrosis and depletion of lymphocytes, attended by infiltration of macrophages. Histopathological changes were also noted in the cerebellum of three fetuses, consisting of necrosis in and depletion of the external germ layer and in the skin and mucous membranes of all four fetuses. The viral antigen was present in cells of the mononuclear phagocyte system, primarily in the lymphoid tissues. The infection was further demonstrated in the affected cerebelli. PMID- 6293689 TI - Field evaluation of test-and-removal and vaccination as control measures for pseudorabies in Missouri swine. AB - Eighteen Missouri swine herds were serologically monitored to determine the efficiency of two methods for the control of pseudorabies. A serum neutralization test-and-removal procedure was effective in ten of ten herds using this method. Vaccination procedures were less reliable. Virus still circulated in five of eight vaccinated herds and a thorough epidemiological evaluation of herd status was impossible. Titers caused by vaccination could not be distinguished from those by natural infection. Vaccination also was carried out in three seronegative herds. Antipseudorabies virus titers in these herds ranged from negative to 1:16 two to three months postvaccination. A majority of the sows in these herds responded with low 1:2 or 1:4 titers following vaccination. A direct comparison of "test and removal" farms and farms that used vaccination was not possible because of the differences between the two groups of farms. PMID- 6293690 TI - Rotaviral antibodies in cow's milk. AB - In the past, it has been reported that neonatal diets made from unheated cow's milk were superior to those made from heated cow's milk. It was observed that piglets were equally protected from rotaviral diarrhea when they were fed diets made from either unheated milk that came from a cow immunized against porcine rotavirus or from a cow that was not immunized. Because of this observation, we examined four pools of "normal" cows' colostrum and 58 samples of "normal" cow's milk for the presence of antibody to rotavirus. All pools of colostrum, collected in four different years, had immunofluorescent antibody titers of 1:100 to rotavirus. Seventy-two percent of the samples of milk were also positive--titer no higher than 1:10. Antibodies to rotavirus were found in cow's milk at a creamery prior to but not after pasteurization. Rotaviral antibodies were detected in one out of eight brands of milk bought at the market--perhaps indicating inadequate pasteurization for this brand. These results support the proposition that, at least in part, unheated milk is superior to heated milk because unheated milk contains antibody to an ubiquitous enteropathogen like rotavirus. PMID- 6293691 TI - Lateral cerebral ventricle and preoptic-anterior hypothalamic area infusion and perfusion of beta-endorphin and ACTH to unrestrained rats: core and surface temperature responses. AB - Both beta-endorphin and ACTH have been found in high concentrations within the hypothalami of mammals and each neuropeptide has been proposed to play a physiological role in regulating body temperature. In an attempt to determine how these peptides may alter thermoregulation, small, microgram concentrations of beta-endorphin and ACTH were injected either into lateral cerebral ventricle (ICV) or directly into the preoptic-anterior hypothalamic area (POAH) or perfused into the POAH of unrestrained rats. Core (rectal) and surface (tail) temperatures were recorded before and after ICV and POAH injection of 1 microgram of beta endorphin or ACTH or perfusion (10 ng/microL) of either neuropeptide. POAH perfusion of naloxone HCl following the neuropeptide perfusion was tested to determine the specificity of the temperature responses. Regardless of the route of central administration, beta-endorphin, in the concentrations used, consistently evoked a hyperthermic core temperature response, that could be antagonized by naloxone. Increased core temperatures may, in part, have been due to peripheral vasoconstriction, as suggested by the decreases seen in tail temperature. The same concentrations of ACTH failed to show any prominent core temperature changes. Results suggest that beta-endorphin is a more potent modulator than ACTH in altering core temperatures of unrestrained rats. Whether beta-endorphin and ACTH act physiologically in an antagonistic manner to maintain a constant body temperature remains to be proven. PMID- 6293692 TI - Radioiodination of p-hydroxyphenylisopropyladenosine: development of a new ligand for adenosine receptors. AB - dl-p-Hydroxyphenylisopropyladenosine has been iodinated with 125I and tested as a ligand of adenosine receptors in membranes from rat brain using a filtration assay. Binding studies using l-[3H]phenylisopropyladenosine as a ligand were carried out in parallel, and the binding of both ligands could be displaced by dl p-hydroxyphenylisopropyladenosine and l-phenylisopropyladenosine in a similar fashion. These data establish the feasibility of using radioiodinated derivatives of adenosine as ligands of suitable types of adenosine receptors. PMID- 6293693 TI - Preformed magnesium hydroxide precipitate for second-step concentration of enteroviruses from drinking and surface waters. AB - A method is described for the second-step concentration of viruses from large volumes of drinking and surface waters. Seeded viruses present in the first eluate, performed with 50 mM glycine buffer, pH 11.5, were adsorbed on a preformed magnesium hydroxide precipitate. After low-speed centrifugation they were desorbed and adjusted to pH 7 with McIlvaine citrate-phosphate buffer. In these experimental conditions 90% of the viruses present in the 300-mL first eluate were reconcentrated in a final volume of 40 mL. The recovery efficiency was independent of either virus concentration or water quality. PMID- 6293694 TI - Adaptive resistance to polymyxin in Pseudomonas aeruginosa due to an outer membrane impermeability mechanism. AB - The isolated outer membrane from cells of a Pseudomonas aeruginosa strain exhibiting adaptive resistance to polymyxin was not affected by polymyxin treatment, as monitored by electron microscopy of negatively stained preparations. This was in sharp contrast with extensive disruption by polymyxin of the outer membranes of the parent polymyxin-sensitive strain and the resistant strain following reversion to greater polymyxin sensitivity. The isolated cytoplasmic membrane of the polymyxin-resistant strain, on the other hand, remained sensitive to the disruptive effects of polymyxin treatment. The permeability characteristics of the resistant strains appear to be altered, as indicated by differences in minimal inhibitory concentrations for a variety of antibiotics between the polymyxin-sensitive and polymyxin-resistant strains. No evidence was found for a polymyxin-inactivating enzyme in osmotic shock fluid from the polymyxin-resistant strain. No evidence for a cytoplasmic membrane repair mechanism was found in the polymyxin-resistant strain. These observations suggest that the mechanism of adaptive polymyxin resistance in this model system is the alteration of the outer membrane so that it excludes polymyxin from reaching the still sensitive cytoplasmic membrane. PMID- 6293695 TI - Characterization of bacteriocin 28 produced by Clostridium perfringens. AB - Bacteriocin 28, produced by Clostridium perfringens, was characterized by gel filtration and sodium dodecyl sulfate - polyacrylamide gel electrophoresis as a glycoprotein with a molecule weight of approximately 100,000. Density gradient centrifugation suggested a lower weight of 84,000. The bacteriocin bound firmly to phenyl-Sepharose CL-4B gel, indicating hydrophobic properties, and elution from this gel with ethylene glycol clearly separated bacteriocin from the alpha and theta toxins of C. perfringens, the latter of which was also hydrophobic. Bacteriocin 28 was immunogenic, inducing neutralizing and precipitating antibodies, and possessed three isoelectric points: 7.37, 7.05, and 5.4. Amino acid and carbohydrate analysis of the active material showed a composition of 15 amino acids and several carbohydrates. The molecule demonstrated instability with increasing purification, and several approaches to purification are described. PMID- 6293696 TI - Spatial distribution of proliferating cells in avian sarcoma virus-induced gliomas. AB - We studied the regional distribution of proliferating tumor cells in five avian sarcoma virus-induced gliomas. The labeling index and spatial distribution of [3H]thymidine (dThd)-labeled tumor cells were determined in serial sections of each tumor with a computer-assisted digitizing system. The density of [3H]dThd labeled cells showed marked regional variation in each tumor, and the ratio of the density of [3H]dThd-labeled cells in tumor periphery to tumor center varied from 0.86 to 1.38. The labeling index generally, but not always, reflected [3H]dThd-labeled cell density. This study indicates that proliferating pools of glioma tumor cells exhibit regional variability in concentration and that the highest numbers of proliferating cells may be predominantly located in central regions of tumor and not in tumor periphery as assumed previously. In all tumors, large numbers of proliferating cells were present in all parts of the tumor. PMID- 6293697 TI - Effect of anthracycline antibiotics on oxygen radical formation in rat heart. AB - This investigation examined the effect of the anthracycline antitumor agents on reactive oxygen metabolism in rat heart. Oxygen radical production by doxorubicin, daunorubicin, and various anthracycline analogues was determined in heart homogenate, sarcoplasmic reticulum, mitochondria, and cytosol, the major sites of cardiac damage by the anthracycline drugs. Superoxide production in heart sarcosomes was significantly increased by anthracycline treatment; for doxorubicin, the reaction appeared to follow saturation kinetics with an apparent Km of 112.62 microM, required NADPH as cofactor, was accompanied by the accumulation of hydrogen peroxide, and probably resulted from the transfer of electrons to molecular oxygen by the doxorubicin semiquinone after reduction of the drug by sarcosomal NADPH:cytochrome P-450 reductase (NADPH:ferricytochrome oxidoreductase, EC 1.6.2.4). Superoxide formation was also significantly enhanced by the anthracycline antibiotics in the mitochondrial fraction. Doxorubicin stimulated mitochondrial superoxide formation in a dose-dependent manner that also appeared to follow saturation kinetics (apparent Km of 454.55 microM); however, drug-related superoxide production by mitochondria required NADH rather than NADPH and was significantly increased in the presence of rotenone, which suggested that the proximal portion of the mitochondrial NADH dehydrogenase complex [NADH:(acceptor) oxidoreductase, EC 1.6.99.3] was responsible for the reduction of doxorubicin at this site. In heart cytosol, anthracycline-induced superoxide formation and oxygen consumption required NADH and were significantly reduced by allopurinol, a potent inhibitor of xanthine oxidase (xanthine:oxygen oxidoreductase, EC 1.2.3.2). Reactive oxygen production was detected in all of our studies despite the presence of both superoxide dismutase (superoxide:superoxide oxidoreductase, EC 1.15.1.1) and glutathione peroxidase (glutathione:hydrogen peroxide oxidoreductase, EC 1.11.1.9) in each cardiac fraction. These results suggest that free radical formation by the anthracycline antitumor agents, which occurs in the same myocardial compartments that are subject to drug-induced tissue injury, may damage the heart by exceeding the oxygen radical detoxifying capacity of cardiac mitochondria and sarcoplasmic reticulum. PMID- 6293698 TI - Systemic excretion of benzo(a)pyrene in the control and microsomally induced rat: the influence of plasma lipoproteins and albumin as carrier molecules. AB - In vitro studies have previously indicated that benzo(a)pyrene distributes primarily into the plasma lipoprotein fraction when incubated with whole plasma. Hydroxylated metabolites of benzo(a)pyrene distribute increasingly into the albumin fraction as the degree of metabolite hydroxylation increases. This report assesses the influence of plasma lipoproteins and albumin as carriers for benzo(a)pyrene on carcinogen excretion in the control and microsomally induced rat. Male Sprague-Dawley rats cannulated in the bile duct received i.v. injections of radiolabeled benzo(a)pyrene noncovalently bound to the very-low density, low-density, or high-density lipoproteins in equimolar amounts. Bile was collected and measured for radioactivity. Cumulative biliary excretions of benzo(a)pyrene complexed with rat lipoproteins were 39.6 +/- 9.7 (S.D.), 24.6 +/- 1.3, and 21.2 +/- 8.8% for very low-density, low-density, and high-density lipoprotein, respectively. Values for excretion of benzo(a)pyrene complexed with rat or human lipoproteins were comparable. These data suggest that the transport molecule can effect a 2-fold difference in benzo(a)pyrene excretion under conditions of the present study. We infer that metabolism of the plasma lipoprotein molecules determines, in part, the extent of benzo(a)pyrene excretion. Cumulative biliary excretions of albumin-bound benzo(a)pyrene, 3 hydroxybenzo(a)pyrene, benzo(a)pyrene 7,8-dihydrodiol, and benzo(a)pyrene-4,5 epoxide were 28.0 +/- 2.7, 39.8 +/- 0.5, 46.9 +/- 2.5, and 49.8 +/- 1.2%, respectively. Thus, excretion increased as the degree of benzo(a)pyrene hydroxylation increased. The effect of microsomal enzyme induction on excretion of lipoprotein-bound benzo(a)pyrene was also assessed. Contrary to expectation, excretion of benzo(a)pyrene bound to the very-low-density, low-density, or high density lipoproteins in Aroclor-induced rats was not greater than that of control animals. Hence, under the conditions of the present study, 60 to 80% of the injected benzo(a)pyrene and 50 to 60% of the injected benzo(a)pyrene metabolites were not excreted immediately in control or microsomally induced animals. This benzo(a)pyrene may represent a carcinogen pool that is slowly excreted. PMID- 6293699 TI - Hormonal and metabolic regulation of human chondrosarcoma in vitro. AB - Prostaglandin A1 has a profound inhibitory effect on uridine incorporation into RNA of normal cartilage whereas N6-monobutyryladenosine 3',5'-cyclic monophosphate is either stimulatory or without an effect. Sera from intact and growth hormone-treated hypophysectomized rats stimulate RNA synthesis but serum from untreated hypophysectomized rats does not. The present study investigated the in vitro regulation of [3H]uridine incorporation into RNA of six human chondrosarcomas to determine if malignant human chondrocytes are under similar metabolic and hormonal regulation. Prostaglandin A1 (25 micrograms/ml) markedly inhibited uridine incorporation in all six tumors (56 to 80%). N6 Monobutyryladenosine 3',5'-cyclic monophosphate (1 mM) inhibited uridine incorporation in five tumors (20 to 50%). Uridine incorporation was stimulated by growth hormone-dependent serum factors in one tumor and by growth hormone independent serum factors in two tumors. Two tumors were more responsive to serum from growth hormone-treated hypophysectomized rats than to serum from intact rats, and one tumor was unresponsive to serum stimulation. The data indicate that: (a) prostaglandin A1 is a very potent inhibitor of RNA synthesis in human chondrosarcomas; (b) N6-monobutyryladenosine 3',5'-cyclic monophosphate affects human chondrosarcomas differently than it does normal cartilage; and (c) responses of human chondrosarcomas to serum growth factors vary among individual tumors. PMID- 6293700 TI - Expression of Friend leukemia virus and spleen focus-forming virus-specific sequences in erythroid bursts and granulocyte-macrophage colonies from spleen and marrow of mice infected with Friend leukemia virus. AB - A large number of studies have been carried out to identify the Friend leukemia virus (FV) target cell(s). In FV-infected mice, the kinetics of "primitive" erythroid burst-forming units (P-BFU-E) is perturbed, and their proliferative rate is enhanced. These results indirectly suggest, but do not prove, that cycling P-BFU-E may serve as FV target. In vitro infection studies showed that normal erythroid colony forming units (CFU-E) and "mature" erythroid burst forming units (M-BFU-E) are targets for FV, while the largely out-of-cycle normal P-BFU-E are not. In an attempt to shed light on these aspects, we have evaluated the expression of viral cytoplasmic RNA sequences in pools of colonies generated by P-BFU-E and granulocyte-macrophage colony forming units (CFU-GM) from spleen and marrow of polycythemic Friend virus (FVP)-infected mice, as measured by liquid hybridization with FVP- or spleen focus-forming polycythemic virus (SFFVp) specific DNA probes. Moreover, similar assays were performed on RNAs derived from whole spleen or bone marrow from mice treated with FVP or the anemic strain of Friend virus (FVA). Control studies were performed on corresponding colonies and whole tissues from normal animals. FVP- and SFFVp-specific sequences are more abundant in RNA extracted from infected spleen as compared to marrow by a 10-fold factor. On the other hand, FVP and SFFVp-specific sequences are expressed at a comparable level in both P-BFU-E- and CFU-GM-derived colonies from spleen or marrow of FVP-treated mice. Since in vitro spread of FVP infection was excluded by control studies with addition in culture of antibody to the viral glycoprotein with a molecular weight of 70,000 (gp70) these results indicate that P-BFU-E and CFU-GM are infected in vivo by FVP. PMID- 6293701 TI - Limitations of DNA histogram analysis by flow cytometry as a method of predicting chemosensitivity in a rat renal cancer model. AB - Analysis of DNA histograms obtained from rat renal cancer cells stained with propidium iodide and submitted to flow cytometry revealed a tumor cell population with prominent 2C and 4C peaks and with usually less than 10% each of S-phase or hyper-4C cells. The presence of an increased proportion of 4C cells was found to depend on the age and size of the tumor nodule. Sampling replicate portions of the same tumor or different tumors of the same size and age frequently revealed highly variable 4C:2C ratios. Treatment of animals bearing this tumor with a single i.p. injection of cyclophosphamide (CY), under conditions known to reduce the tumor burden by 80% within 1 week, or with 5-fluorouracil (FUra), which is ineffective against this tumor, in many cases did not yield changes in DNA histograms that permitted one to distinguish the effective drug. Either no marked difference in histogram shape occurred after therapy, or FUra induced more striking differences in cell cycle position than did CY. In tumor generations with greater than 15% S-phase cells, treatment with CY resulted in multiple effects on DNA histograms. These included detecting increased numbers of moribund cells (hypo-2C), a decrease in 4C cells, and an increase in hyper-4C cells. These changes did not occur with the ineffective agent FUra. The tumors grown in vitro show no evidence of replication of 4C cells. The DNA histograms of late-log-phase cultures have a major 2C peak and a minor S plus G2 hump. Since neither the untreated tumor in vivo nor that grown in vitro has a major hyper-4C cell population, it is probable that the tumor stem cells are chiefly 2C (diploid hyperdiploid). Treatment in vitro of late-log-phase cultures with CY or FUra produces DNA histograms which permit identification of the effective agent. After CY, a major part of the cell population was hypo-2C (moribund) cells. PMID- 6293702 TI - Effects of dibutyryl cyclic adenosine 3':5'-monophosphate on the growth of cultured human small-cell lung carcinoma and the specific cellular activity of L dopa decarboxylase. AB - High activity of L-dopa decarboxylase separates small (oat)-cell from non-small cell lung cancer in cell culture. The present study investigates relationships between the specific cellular activity of this enzyme and: (a) cell growth kinetics of an established line (O-H-1) of human small cell lung carcinoma, and (b) responses of these cells to treatment with cyclic adenosine 3':5' monophosphate and sodium butyrate. The O-H-1 cells, as for most other established small-cell lines, grow as suspended cell aggregates. During growth, the specific cellular activity of L-dopa decarboxylase parallels levels for [3H]thymidine labeling index and the ratio of cells in G2-M to those in G1-G0 phases of the cell cycle. Each of these parameters is 2- to 3-fold higher during exponential versus stationary growth. Continuous treatment with dibutyryl cyclic adenosine 3':5'-monophosphate (dcAMP; 0.1 or 1 mM) and 1 mM theophylline produces simultaneous cessation of cell growth and an increase in cellular L-dopa decarboxylase activity. During this period, analyses of DNA histograms reveal an increase in the number of cells in the G2-M phase; the rate of increase in the ratio of G2-M to G1-Go cells paralleled the rate of increase in specific activity of the enzyme. The effects of the dcAMP were promptly reversible; release of the apparent G2-M block preceded regrowth of the cells and was accompanied by a return of L-dopa decarboxylase activity to base-line levels. The changes in enzyme activity were specific for cyclic adenosine 3':5'-monophosphate; another cyclic adenosine 3':5'-monophosphate analogue, 8-bromo adenosine cyclic 3':5' monophosphate yielded similar increases in L-dopa decarboxylase to those seen with dcAMP, while 0.01 to 1 mM butyrate alone produced the inhibition of cell growth but no changes in specific activity of L-dopa decarboxylase or percentage of cells in the different phases of the cell cycle. We conclude that the specific activity of L-dopa decarboxylase, a key neuroendocrine marker for cultured small cell lung carcinoma, is highest during proliferative growth and/or when these cells are in the G2M phase of the cell cycle. The differential effects of dcAMP and sodium butyrate offer potential for exploring neuroendocrine differentiation in this important lung cancer and related endocrine neoplasms. PMID- 6293703 TI - Immunological detection of carcinogen-modified DNA fragments after in vivo modification of cellular and viral chromatin. AB - Antibodies specific for DNA modified by (+/-)-trans-7, 8-dihydrobenzo(a)pyrene 7,8-diol-9, 10-epoxide have been used to quantitate the relative modification level in fragments derived from pBR322 DNA from cellular DNA and in the coding and noncoding strands of simian virus 40 DNA. DNA fragments with a covalent molar modification level ranging from less than 1 to over 200 are resolved by agarose gel electrophoresis and transferred to diazobenzyloxymethyl cellulose paper. The paper is incubated with antibodies specific to carcinogen-modified DNA, and the location of the antibody is visualized by autoradiography after incubation with 125I-protein A. The binding of antibodies is directly proportional to the level of DNA modification. Using this technique, we find that linker DNA is about 2.5- to 3-fold more accessible to (+/-)-trans-7,8-dehydrobenzo(a)pyrene-7, 8-diol-9, 10-epoxide than nucleosomal core DNA and that under in vivo conditions the coding and noncoding strands of the simian virus 40 chromosome are equally accessible to trans-7,8-dihydrobenzo[a]pyrene-7,8-diol-9, 10-epoxide. The approach described allows assessment of the relative level of modification in any DNA sequence which can be subjected to gel electrophoresis. PMID- 6293704 TI - Enhancement of antitumor activity of ascorbate against Ehrlich ascites tumor cells by the copper:glycylglycylhistidine complex. AB - Ascorbate in an aqueous solution is easily oxidized by molecular oxygen in the presence of cupric ion, thus producing reactive oxygen species and exhibiting cytotoxicity. In order to increase the antitumor activity of ascorbate, we used the innocuous form of cupric ion complexed with glycylglycylhistidine, a tripeptide designed to mimic the specific Cu(II) transport site of albumin molecule. Although this square planar copper:glycylglycylhistidine complex did not significantly oxidize ascorbate at pH 7.4, it killed Ehrlich ascites tumor cells in vitro in a high concentration of ascorbate. The injections of large doses of ascorbate together with copper: glycylglycylhistidine prolonged the life span of mice inoculated i.p. with Ehrlich tumor cells. The target specificity against tumor cells was primarily attributable to their high peptide-cleaving activity. PMID- 6293705 TI - Selective cytotoxicity of purified homologues of tunicamycin on transformed BALB/3T3 fibroblasts. AB - The selective cytotoxicity of tunicamycin homologues against SV40-transformed 3T3 cells (SV40-3T3) was examined. Incubation of 3T3 or virally transformed 3T3 cells with four different homologues (A1, A2, B1, and B2 at 0.1 to 0.25 microgram/ml) caused detachment and death of transformed cells after 1 to 3 days, while the nontransformed cells were almost unaffected. Cytotoxicity against nontransformed cells occurred only when higher doses (at least 5-fold) of A2-, B1-, and B2 tunicamycins were used. In contrast, these homologues inhibited proliferation of 3T3 cells, even when doses of 0.5 microgram/ml were used. These cytotoxic effects are dose dependent, and maximal cytotoxicity of each homologue is achieved at a different concentration in each cell type. These results indicate that tunicamycin homologues have selective cytotoxicity against transformed cells. Incorporation of [3H]mannose into acid-precipitable macromolecules synthesized by transformed cells was strongly inhibited (70 to 75%) by A1- and B2-tunicamycins at 0.01 to 0.05 microgram/ml, while incorporation by 3T3 cells was not affected. At higher concentrations of the above tunicamycins (0.5 to 1 microgram/ml), [3H]mannose incorporation by both 3T3 and SV40-3T3 cells was inhibited more than 95%. In contrast, the effect of these tunicamycin homologues on protein synthesis in 3T3 and SV40-3T3 fibroblasts was less pronounced since the incorporation of amino acids was inhibited by approximately 20%. Very little inhibition of amino acid incorporation occurred when 3T3 or SV40-3T3 cells were treated with B2 tunicamycin. However, A1-tunicamycin inhibited [3H]proline incorporation and slightly increased [3H]tyrosine incorporation into cell layers of 3T3 cells. Examination of secreted proteins synthesized by these cells on sodium dodecyl sulfate:polyacrylamide gel electrophoresis revealed that both 3T3 and SV40-3T3 cells treated with homologues produced partially glycosylated macromolecules, such as procollagen and fibronectin, and failed to convert procollagen to collagen. Tunicamycin homologues also inhibited the N-acetylglucosamine-1 phosphate transferase activity found in microsomes prepared from 3T3 and virally transformed 3T3 fibroblasts. The data presented indicate that the cytotoxic activity of purified homologues of tunicamycin against transformed fibroblasts might be due to the selective inhibition of glycosylation and to the differences in the membrane solubilities of the homologues. PMID- 6293706 TI - Antibodies against three purified proteins of the human type C retrovirus, human T-cell leukemia-lymphoma virus, in adult T-cell leukemia-lymphoma patients and healthy Blacks from the Caribbean. AB - Six black patients of Caribbean origin with adult T-cell leukemia-lymphoma, 18 of their healthy family members and relatives, and 337 healthy black individuals from the Caribbean were investigated for the presence of serum antibodies against human T-cell leukemia-lymphoma virus (HTLV). Three distinct structural proteins of this virus with molecular weights of 24,000, 19,000, and 15,000 were purified, radiolabeled, and used in radioimmune precipitation assays. Five of the patients, three of the family members (two of them spouses), and 11 of the normals had specific antibodies against at least the proteins with molecular weights of 24,000 and 19,000. High antibody titers against these proteins were often associated with antibodies against the protein with a molecular weight of 15,000. In all cases, antibody titers against this protein were considerably lower than those against the proteins with molecular weights of 24,000 and 19,000. HTLV is highly associated with Caribbean adult T-cell leukemia-lymphoma and is also endemic among the normal Caribbean population. By comparison of the frequencies of anti-HTLV positives among family members of patients and the normal population, we conclude that infection by HTLV occurs in a horizontal way and at least in the West Indian black and Japanese population probably without a requirement for uncommon genetic factors. PMID- 6293707 TI - An overall review on prenylamine mechanisms of action in experimental models of myocardial damage. AB - In this review we present the effects of a well-known antianginal drug, prenylamine (PNL), in experimental models of acute myocardial damage induced by a beta-agonistic drug, isoproterenol (ISP), in several trials conducted in our laboratory in both rats (n = 204) and monkeys (n = 26). PNL significantly inhibited ISP-induced lesions, protecting the majority of animals studied. Studies dealing with the site of action of the drug, such as 45Ca, 3H-PNL and 3H ISP trials, showed a clear membrane effect slowing down Ca transport. Correlation between ECG (inhibition of ST depression after ISP) and pathological findings in monkeys was also obtained in one of our experiments. These series of assays were useful in obtaining a more complete idea of activity and site of action of the drug. It seems that, in acute models, PNL acts as a calcium antagonistic drug rather than an adrenergic moderator. PMID- 6293708 TI - Role of cyclic nucleotides in heart metabolism. PMID- 6293709 TI - Detection of severity of coronary artery disease by the ST segment/heart rate relationship in patients on beta-blocker therapy. AB - Many patients presenting with anginal pain are on beta-blocker therapy and it has been suggested that noninvasive exercise tests for coronary artery disease are adversely affected by such therapy. Recently a new exercise test has been introduced claiming the ability accurately to detect the presence and severity of coronary heart disease in patients with anginal pain; the claim was based on results obtained in patients not undergoing beta-blocker therapy. Therefore using this test, the maximal rate of progression of ST segment depression relative to increases in heart rate during exercise (maximal ST/HR slope) as an index of the severity of myocardial ischaemia, 60 patients on beta-blocker therapy were studied and the results compared with results of coronary arteriography independently obtained. There was a complete agreement, without false results, between the maximal ST/HR slope and the number of significantly diseased coronary arteries. In 21 of these 60 patients the maximal ST/HR slopes obtained before and after beta-blockade were compared; the maximal ST/HR slopes after beta-blockade were not different from those before. It is concluded, that the maximal SR/HR slope can be used reliably to detect the pressure and the severity of coronary heart disease in patients with angina pectoris who are already on beta-blocker therapy. PMID- 6293710 TI - Effects of physical training on beta-adrenergic receptors in rat myocardial tissue. AB - A diminished sympathetic activity has been related to training bradycardia seen at rest and during exercise. In order to evaluate if changes in heart adrenergic receptors can be one of the mechanisms by which the sympathetic responsiveness could be decreased by physical training, the number and affinity of beta adrenergic receptors were determined in heart ventricular tissue of rats submitted to a 10-week running programme. Binding studies were done at different concentrations of (-)[3H] dihydroalprenolol (DHA) (0.5 to 14.4 nmol X litre-1) with ventricular membrane preparations from control and trained rats. Direct linear plot analysis revealed that physical training reduced the total number (1933 +/- 192 vs 2922 +/- 211 fmol X ventricles-1; P less than 0.01) density of beta-adrenergic receptors expressed either as fmol X mg-1 of membrane protein (34 +/- 3 vs 43 +/- 3; P less than 0.05) or as fmol X g-1 ventricle (1740 +/- 170 vs 2308 +/- 155; P less than 0.05). There was no significant change in the dissociation constant (3.11 +/- 0.14 vs 4.08 +/- 0.51 nmol X litre-1; P greater than 0.05). Basal plasma noradrenaline levels were not affected by training (116 +/- 18 vs 101 +/- 14 pg X cm-3; P greater than 0.10); however the adrenaline values were significantly higher in trained rats (91 +/- 16 vs 47 +/- 7 pg X cm 3; P less than 0.05). These data indicate that physical training induces changes at the level of beta-adrenergic receptors and this may partly explain the bradycardia seen in trained subjects and animals. PMID- 6293711 TI - The surgical applications of cefotaxime. Proceedings of a symposium. Hospital for Joint Diseases, Orthopaedic Institute, New York, June 22, 1982. PMID- 6293712 TI - Clinical evaluation of cefotaxime versus gentamicin plus clindamycin in the treatment of polymicrobial peritonitis. AB - One hundred fifty-one patients with presumed aerobic-anaerobic mixed peritoneal infections were treated in a prospective, randomized trial with either cefotaxime alone (76) or the combination of gentamicin-clindamycin (75). Primary and complicating foci of sepsis were cultured for both aerobic and anaerobic pathogen identification and antibiotic susceptibility. In vitro aerobic disk sensitivities (114 isolates) to cefotaxime were 82% and to gentamicin, 88%; anaerobic agar diffusion sensitivities (227 isolates) to cefotaxime were 87% and to clindamycin, 98%. Only enterococci and Pseudomonas sp were consistently resistant to cefotaxime. Infection was eliminated in 82% of those treated with cefotaxime and in 87% of those treated with the gentamicin-clindamycin combination, yet sepsis recurred in 11% of those treated with cefotaxime and in 13% for those given gentamicin-clindamycin. Five patients (7%) demonstrated nephrotoxicity for gentamicin. (Serum creatinine increased greater than 1.5 mg/100 ml over pretreatment levels.) Otherwise, incidence and severity of adverse reactions were identical for the two groups and consisted primarily of phlebitis and diarrhea. One patient in each treatment group died of uncontrolled sepsis. Although results suggested a laboratory superiority of gentamicin-clindamycin, there was a clinical equality in therapeutic benefit and a greater safety following the use of cefotaxime alone. PMID- 6293713 TI - Cefotaxime treatment in patients with osteomyelitis and septic arthritis: a multicenter study. AB - The effectiveness of parenteral cefotaxime in the treatment of adults with acute septic arthritis or acute or chronic osteomyelitis was evaluated in a multicenter trial. The drug was given to 47 patients admitted to the University of Texas Medical Branch Hospitals or Hahnemann Medical College and Hospital (UT-H study) and to 40 patients in other medical centers using an identical protocol. In the UT-H study, cefotaxime was effective in 15 of 16 patients (94%) with acute osteomyelitis, in 24 of 27 patients (89%) with chronic osteomyelitis, and in four of four patients (100%) with acute septic arthritis. In the multicenter study, the success rates were as follows: acute osteomyelitis, six of six (100%); chronic osteomyelitis, 14 of 19 (74%); and septic arthritis, 12 of 15 (80%). The antibiotic was well tolerated in most patients. The most serious side effect was significant neutropenia, which occurred in three patients. Cefotaxime appears to be a clinically useful, broad-spectrum antibiotic for bone and joint infections. PMID- 6293714 TI - Cefotaxime treatment of skin and skin structure infections: a multicenter study. AB - The efficacy, safety, and tolerability of cefotaxime, a new parenteral cephalosporin resistant to beta-lactamase, were evaluated in a multicenter open trial. The study population comprised 594 hospitalized patients with infections of the skin or subcutaneous tissue. Of these, 409 patients fulfilled the protocol requirements for assessment of clinical efficacy. Usual dosages of cefotaxime were in the range of 1.5 to 12 gm/day for 5 to 85 days. Staphylococcus aureus was the most frequent gram-positive organism; Proteus mirabilis, Escherichia coli, and Pseudomonas aeruginosa, the most frequent gram-negative bacilli; and Peptococcus sp, the most frequent anaerobic organism isolated. Of the 409 evaluable patients, 382 (93.4%) were clinically cured. Bacteriological cures were obtained in 316 of 372 (84.9%) patients. Reactions to cefotaxime included asymptomatic eosinophilia, rash, drug fever, phlebitis, and elevated hepatic enzyme levels. All reactions were transient, and drug therapy had to be terminated in only eight patients. Cefotaxime proved effective for treating a variety of skin and soft tissue infections. The lack of serious toxicity and adverse reactions associated with cefotaxime, together with its broad antimicrobial spectrum, makes it a suitable substitute for aminoglycosides in certain clinical settings. PMID- 6293715 TI - Clinical comparison of cefotaxime versus the combination of gentamicin plus clindamycin in the treatment of polymicrobial soft-tissue surgical sepsis. AB - The safety and efficacy of cefotaxime versus a combination of gentamicin and clindamycin were compared in a prospective, randomized study of 98 surgical patients with polymicrobial soft-tissue infection or septicemia. Forty-nine patients received cefotaxime (20 mg/kg every six hours), and 49 received gentamicin (1 mg/kg every eight hours) plus clindamycin (5 mg/kg every six hours); all drugs were given intravenously. Overall, there was no statistical difference in clinical response to the two regimens, infection being eliminated in 73% of the patients treated with cefotaxime and 71% of those given gentamicin plus clindamycin. Adverse effects were mild and self-limited in both treatment groups, although three patients treated with gentamicin plus clindamycin experienced some loss of renal function. Most aerobic gram-negative rods were sensitive to both cefotaxime and gentamicin, but anaerobes were slightly more sensitive to clindamycin than to cefotaxime. Cefotaxime appeared to be at least as effective as gentamicin plus clindamycin in the treatment of polymicrobial soft-tissue infections and septicemia, and, in light of the loss of renal function associated with the gentamicin-clindamycin regimen, somewhat safer. The high failure rate among patients on both regimens with septicemia of unknown origin (five of the nine treated with cefotaxime and two of the four treated with gentamicin and clindamycin), however, indicates the critical role of surgical management in the treatment of polymicrobial soft-tissue sepsis. PMID- 6293716 TI - A prospective randomized study of prophylactic mannitol (10%)-neomycin-cefotaxime therapy in patients undergoing elective colonic and rectal surgery. AB - A prospective randomized trial was performed to assess the effectiveness of 10% mannitol, oral neomycin, and intravenous cefotaxime in preventing septic complications following elective colonic and rectal surgery in 99 patients. In each of the three regimens, 10% mannitol and oral neomycin were given preoperatively. In regimen 1 (34 patients) intravenous cefotaxime was given perioperatively. In regimen 2 (33 patients) cefotaxime was given perioperatively and for 24 hours postoperatively. Regimen 3 (32 patients) was the same as regimen 2 except that cefazolin sodium was substituted for cefotaxime. Septic complications developed in one patient each on regimens 1 and 3. There were no infections in the patients on regimen 2. This study demonstrates that bowel preparation with 10% mannitol and oral neomycin, coupled with a short perioperative course of cefotaxime or cefazolin sodium, is effective in controlling the septic complications of elective colorectal surgery, and the combinations are simple, safe, and cost effective. PMID- 6293717 TI - Antibiotic prophylaxis in genitourinary surgery: a comparison of cefotaxime and cefazolin. AB - In a prospective, single-blind study, the efficacy of cefotaxime and cefazolin in the prevention of postoperative infection was compared in 133 patients admitted for elective genitourinary surgery (over half of whom were undergoing transurethral resection). In regimen 1, cefotaxime was given perioperatively (before, during, and up to two hours after surgery); in regimen 2, cefotaxime was given perioperatively and for 24 hours postoperatively; and in regimen 3, cefazolin was given perioperatively and for 24 hours postoperatively. All doses equaled 1 gm and were given either intramuscularly or by intravenous bolus. All patients had negative urine cultures preoperatively. The criterion for absence of infection was a negative urine culture (less than 10(5) organisms/ml) after surgery. At the final evaluation, 113 of the 133 patients (85%) were free of infection--39 of 45 (86.7%) on regimen 1, 40 of 45 (88.9%) on regimen 2, and 34 of 43 (79.1%) on regimen 3. All 20 patients who had postoperative infections had been catheterized for at least three days. Only 12 patients had temperatures greater than or equal to 101 F two days after surgery, and none had been clinically septic. No side effects other than a mild rash and itching in one patient on regimen 2 were reported. Both cefotaxime regimens were slightly more effective, bacteriologically and clinically, than the standard prophylactic cefazolin regimen. It is concluded that cefotaxime given perioperatively (up to two hours postoperatively) and/or up to 24 hours postoperatively effectively prevents postoperative complications associated with genitourinary surgery. PMID- 6293718 TI - Short-term cephalosporin prophylaxis in transurethral surgery. AB - The effects of three short-term prophylactic cephalosporin regimens were compared in a prospective, randomized, double-blind study. One hundred sixteen men, with sterile urine preoperatively, underwent transurethral surgery (resection of the prostate or bladder neck, 73; resection or fulguration of bladder tumors, 38; or internal urethrotomy, 5). One group received 1 gm of cefotaxime parenterally 30 to 90 minutes preoperatively, every 60 to 90 minutes during surgery, and immediately postoperatively. In the other two groups, 1 gm of cefotaxime or cefazolin was administered according to the regimen used in the first group and also every eight hours for 24 hours postoperatively. All three prophylactic regimens significantly lowered the incidence of infection three and five to seven days postoperatively, the incidence being approximately 5% as compared with an incidence of about 40% found in an earlier study in which a placebo was used. The results with the three regimens were not significantly different. Our data showed that to prevent postoperative urinary tract infection, prophylactic antibiotics need only be administered immediately perioperatively in uninfected patients who undergo transurethral surgery; longer prophylaxis is unnecessary. PMID- 6293719 TI - Comparative efficacy of cefotaxime and cefazolin as prophylaxis against infections following elective hysterectomy. AB - The safety and efficacy of cefotaxime and cefazolin as prophylaxis against posthysterectomy infections were compared in a prospective, randomized study. A total of 118 women undergoing elective vaginal or abdominal hysterectomy were randomly assigned to receive either (1) cefotaxime perioperatively, (2) cefotaxime perioperatively and 24 hours postoperatively, or (3) cefazolin perioperatively and 24 hours postoperatively. In all regimens, the initial antibiotic dose was given intramuscularly and subsequent doses were given intravenously. No postoperative pelvic or wound infections developed in patients in any of the three study groups during the hospital stay or in the 30-day follow up period. No side effects or changes in laboratory test values attributable to the antibiotics were noted. The results of the study indicate that (1) standard one-day regimens of cefotaxime and cefazolin are equally safe and effective as prophylaxis against posthysterectomy infections, and (2) a brief, three-dose perioperative course of cefotaxime is as effective as a standard one-day regimen of either cefotaxime or cefazolin and has the advantages of reduced cost and greater convenience. PMID- 6293720 TI - Comparison of cefotaxime with cefazolin for prophylaxis of vaginal or abdominal hysterectomy. AB - One hundred fourteen women undergoing vaginal hysterectomy (n = 63) or abdominal hysterectomy (n = 51) were randomly allocated to one of three groups: regimen 1- cefotaxime perioperatively; regimen 2--cefotaxime perioperatively and for 24 hours postoperatively; or regimen 3--cefazolin perioperatively and for 24 hours postoperatively. Febrile morbidity was evaluated with respect to antibiotics, number of doses, type of procedure, operative time, estimated blood loss, and wound bacteriology. Of the patients who underwent vaginal hysterectomy, febrile morbidity occurred in three of 24 on regimen 1, one of 23 on regimen 2, and zero of 16 on regimen 3. Of those patients who underwent abdominal hysterectomy, febrile morbidity occurred in three of 18 on regimen 1, zero of 19 on regimen 2, and two of 14 on regimen 3. There were no significant differences among regimens in the incidence of febrile morbidity after vaginal or abdominal hysterectomy. Findings indicate, however, that febrile morbidity correlates with the type of procedure rather than with any of the other variables studied. Return of vaginal flora (aerobes, anaerobes, or mixed) was similar with all regimens. Cefotaxime was found to be as effective as cefazolin in reducing febrile morbidity. PMID- 6293721 TI - Cefotaxime, cefazolin, or ampicillin prophylaxis of febrile morbidity in emergency cesarean sections. AB - Cefotaxime, a new third-generation cephalosporin, was compared with ampicillin and cefazolin in a randomized double-blind trial to evaluate the efficacy of antibiotic prophylaxis of febrile morbidity associated with emergency cesarean sections. A 1-gm intravenous dose of one of the three antibiotics was given by bolus injection immediately after clamping of the umbilical cord and six and 12 hours later. All patients were in labor with membranes ruptured and had a temperature less than or equal to 37.8 C, and none had a history of penicillin or cephalosporin allergy. A total of 195 women were entered into the trial. Initially, the study included a placebo control group which was switched to ampicillin after 30 patients. Of the 188 evaluable patients, 51 of 59 (86.5%) ampicillin recipients, 59 of 67 (88.1%) cefazolin recipients, 48 of 55 (87.3%) cefotaxime recipients, and two of seven (28.5%) placebo recipients had uneventful postoperative courses. During the study, an additional 39 women who were in labor with ruptured membranes but who were allergic to penicillin or who declined antibiotic prophylaxis were classified as untreated patients and observed for postoperative complications. Standard febrile morbidity, primarily related to endometritis or wound infections, occurred in 6 of 59 (10.1%) ampicillin, 5 of 67 (7.5%) cefazolin, 5 of 55 (9.1%) cefotaxime, and 18 (40.0%) of placebo or untreated patients. Cefotaxime, cefazolin, and ampicillin were equally effective in reducing febrile morbidity in emergency cesarean sections. PMID- 6293722 TI - Update of safety of cefotaxime. AB - Adverse reactions to various antibacterial drugs were compared in a review of the following: multiple-dose studies of cefotaxime (n = 3,463), cefazolin (n = 554), the combination gentamicin-clindamycin (n = 163), and cefoxitin (n = 18); prophylactic studies of cefotaxime (n = 300) and cefazolin (n = 149); and single dose studies of cefotaxime (n = 314) and penicillin G procaine (n - 265). The demographic and background characteristics of the groups were similar. Results of extensive pretreatment and posttreatment laboratory tests, measures of vital signs, and physical examinations revealed no clinically important intergroup differences. In the multiple-dose studies, side effects were reported in 9.8% of the cefotaxime, 3.8% of the cefazolin, 17.2% of the gentamicin-clindamycin, and 16.7% of the cefoxitin patients. The most frequent side effects were reactions at the injection site, of the skin and appendages, and of the digestive and urogenital systems, the only significant difference being fewer injection-site reactions in the cefazolin group than in the other three groups. In the prophylactic studies one cefotaxime patient reported rash and pruritus. In the single-dose studies, side effects were reported in 1.6% of the cefotaxime and 4.2% of the penicillin patients. Side effects sufficiently severe to warrant drug discontinuation were reported in 2.1% of the cefotaxime, 0.7% of the cefazolin, 1.8% of the gentamicin-clindamycin, and in none of the cefoxitin patients. Posttreatment prolongation of prothrombin time was found in one cefotaxime patient, whose pretreatment value was also abnormal, and in two gentamicin clindamycin patients. No patient deaths were attributed to any of the drugs. PMID- 6293723 TI - Presence in mouse DNA of repeated sequences related to small regions of the genome of Gross/AKR murine leukemia virus. PMID- 6293725 TI - Targeting of lactoperoxidase to phosphomannosyl-specific receptors on fibroblasts. PMID- 6293726 TI - [Antibodies to the Epstein-Barr virus in pregnant women with suspected teratogenic virus infection]. PMID- 6293724 TI - Immunocytochemical study of Wulzen's cone of the bovine pituitary. AB - The Wulzen's cone of the bovine adenohypophysis presents a variable development and general arrangement. It is joined to the pars intermedia with no intervening connective tissue. It is covered by a single layer of cubical cell epithelium on the side of the hypophysial cleft. Immunofluorescence reveals the presence of different glandular cell types. The most abundant cells are those demonstrated by an anti-oPRL antibody and are either isolated or clustered. Other cells react with anti-hGH, anti-bLH, anti-beta aLH or anti-beta hTSH antibodies. Some cells react simultaneously with anti-beta MSH, anti-1-24ACTH, anti-17-39ACTH, anti-beta LPH and anti-beta endorphin antibodies. Cell types other than the numerous prolactin cells appear only as isolated elements. We did not observe cells reacting with anti-leu-enkephalin, anti-met-enkephalin or anti-calcitonin antibodies either in the Wulzen's cone or in the pars distalis or pars intermedia. PMID- 6293727 TI - Adamantyl nitroxide: a spin label for probing membrane surfaces. AB - In an effort to understand more about the perturbing properties of adamantane like molecules on biological membranes, the spin probe adamantyl nitroxide (2,2' dimethyl-5-adamantyl oxazolidine-N-oxyl) was synthesized, purified and characterized. Electron paramagnetic resonance (EPR) spectra were then obtained from 1:50 and 1:200 mixtures of adamantyl nitroxide with dipalmitoyl and dipalmityl phosphatidylcholine multibilayers. Above the phase transition temperature of these lipids (41 degrees C for dipalmitoyl phosphatidylcholine and 43 degrees C for dipalmityl phosphatidylcholine) the spectra of adamantyl nitroxide are similar to control spectra obtained in liquid oleic acid. Below the phase transition temperatures, however, spectral differences were observed depending on: (1) the concentration of the spin probe in the lipid; (2) the linkage between the polar head group and the hydrocarbon tails of the phospholipid; (3) the temperature of the sample. Partitioning of adamantyl nitroxide between the aqueous and hydrocarbon phases of the sample is most prominent at probe-to-lipid ratios of 1:200 and at temperatures below the pre transition temperature of the lipid (around 33 degrees C). Computer simulations of the above results, as well as additional experiments performed at 35 GHz, show that the results arise from true partitioning and not from asymmetric probe motion. Two conclusive results of these experiments are that spectra of adamantyl nitroxide in phospholipid multibilayers are sensitive to probe concentration and to the physical characteristics of the phospholipid which they probe. The spectral differences which arise when adamantyl nitroxide is used with ether- and ester-linked phospholipids indicate that it is a sensitive probe of membrane surfaces. Employment of this molecule in membrane research should prove to be useful in obtaining additional information about membrane surface events. PMID- 6293728 TI - [Review of 17 cases of human hydatiosis published in West Africa. Apropos of the 4th Ivory Coast case]. AB - Seventeen cases of echinococcus cysts in West Africa has been published up today. All concern Black Africans. Ten are located in the lungs, two in the liver. Want of any animal or human epidemiological studies doesn't permit to find out the reasons of this scarceness. PMID- 6293729 TI - [Traps in orbitotomy]. PMID- 6293730 TI - Acute sodium fluoride toxicity in the rat kidney. AB - Acute sodium fluoride toxicity was studied histologically and histochemically in the rat kidney. The doses given subcutaneously by a single injection were 10, 12.5 and 15 mg/100 gm of body weight and the duration after the administration were 30 minutes, and one, 3, 5 and 7 days. Histologically, coagulation necrosis of the proximal tubules was found at 1 day and the injury was most striking at 3 days. Regeneration was almost complete at 7 days. Among the enzymes studied, e.g., alkaline phosphatase, acid phosphatase, 5'-nucleotidase, adenosine triphosphatase and succinic dehydrogenase, and acid phosphatase of the tubular epithelium was the most severely impaired. PMID- 6293731 TI - The effects of dimercaptosuccinic acid and other chelating agents on the retention of platinum in the rat kidney after treatment with cisplatin. AB - The ability of three chelating agents, dimercaptopropanesulphonate (DMPS), dimercaptosuccinic acid (DMSA), and diethylenetriaminepentaacetic acid (DPTA), to reduce the retention of platinum in the kidneys was tested in male Sprague-Dawley rats treated 24 h previously with Cisplatin. DMPS and DMSA, when given as four daily doses of 1 mMol/kg, increased the urinary excretion from 10.1% +/- 1.2% to 13.6% +/- 1.3% and 13.5% +/- 2.6%, respectively, but only DMSA caused a small but significant reduction in the kidney platinum content. DTPA was totally ineffective. It is concluded that none of these agents is likely to be useful for the reduction of Cisplatin-induced renal toxicity in the clinical situation. PMID- 6293732 TI - Binding of benzo[a]pyrene and intracellular transport of a bound electrophilic benzo[a]pyrene metabolite by lipoproteins. AB - Human serum lipoproteins were isolated by means of size exclusion h.p.l.c. Non covalent uptake of [3H]benzo[a]pyrene was quantitated for fractions collected from the effluent of a liquid chromatographic separation of human serum, and was found to directly correlate with the lipoprotein concentration. An electrophilic benzo[a]pyrene metabolite, [3H]trans 7,8-dihydrodiol-9,10-epoxybenzo[a]pyrene, non-covalently associated with low density lipoproteins was transferred to human lymphocytes in vitro and bound acid-precipitable nucleic acids of the lymphocytes as a function of time. Benzo[a]pyrene metabolite binding to lymphocyte DNA was demonstrated by means of CsCl density gradient analysis. Non-mitogen-stimulated lymphocytes exposed to very low concentrations of carcinogen in the presence of low density lipoprotein demonstrated [3H]thymidine incorporation; without the concomitant addition of low density lipoprotein the low concentrations of carcinogen did not stimulate [3H]thymidine incorporation. PMID- 6293733 TI - DNA in proximity to the site of replication is preferentially alkylated in S phase 10T1/2 cells treated with N-methyl-N-nitroso-urea. AB - Replicating DNA is more susceptible to modification by N-methyl-N-nitrosourea (MNU), a spontaneously active methylating agent, than bulk DNA. This conclusion is supported by results from two different experimental approaches. First, synchronized C3H 10T1/2 clone 8 cells were treated in S phase with MNU and DNA replicated during the period of treatment was separated from bulk DNA. This was done by digesting the purified DNA with restriction enzymes and retaining the replication fork-associated DNA in nitro-cellulose filters. Second, synchronized C3H 10T1/2 clone 8 cells were exposed to 5-bromodeoxyuridine and [3H]MNU and the density-labelled, replicated DNA was separated in CsCl gradients. Both methods show 2.6 to 5.0 times more [3H]methyl adducts per nucleotide residue associated with replicating DNA than that expected from random methylation. These experiments were done at low MNU concentrations (0.018-0.115 mM) that did not cause any detectable inhibition of DNA synthesis or stimulation of repair replication in 10T1/2 cells. PMID- 6293734 TI - Acceptors for the poly ADP-ribosylation modification of chromatin structure are altered by carcinogen-induced DNA damage. AB - Poly(ADP-Rib) polymerase is activated by strand breaks in DNA and appears to play an important role in DNA repair. The enzyme catalyses the poly ADP-ribosylation of histones and non-histone proteins, yet the contribution of these major alterations in chromatin composition have, as yet, not been critically evaluated with regard to DNA strand breaks. In the present study, the effects of N-methyl-N nitrosourea (MNU) upon the poly ADP-ribosylation of nuclear protein acceptors have been identified and quantified at the oligonucleosomal level of chromatin. Treatment of HeLa cells with MNU (4.5 mM) for 1 h resulted in a reduction in the cellular NAD pool (30%), a 2-3 fold stimulation of poly ADP-ribosylation in isolated nuclei and in isolated oligonucleosomes. Of acceptors modified, the automodification of the polymerase was stimulated at least 3-fold. Analysis of the acid-soluble acceptors showed a stimulation in the modification of the core histones and a 2-fold increase in histone H1 poly ADP-ribosylation. This modification causes a novel crosslinking of the latter histone, and this has been studied as it relates to DNA strand breaks in the present work. In vivo treatment with MNU resulted in the synthesis of longer chain or more complex polymer species at the expense of the shorter chained ADP-ribose moieties. PMID- 6293736 TI - Geno- and cytotoxicity of nitrosamines, aflatoxin B1, and benzo[a]-pyrene in continuous cultures of rat hepatoma cells. AB - Two closely related hepatoma cell lines were examined for their response to carcinogens requiring metabolic activation: H5, a dedifferentiated line expressing cytochrome P-448-dependent monooxygenase(s); and HF1-4, a differentiated line which also expresses cytochrome P-450-dependent monooxygenase(s). The hepatocarcinogens dimethyl- and diethylnitrosamine and aflatoxin B1, preferred substrates for cytochrome P-450-dependent monooxygenase(s), and the non-hepatocarcinogen benzo[a]pyrene, which is preferentially metabolized by cytochrome P-448-dependent monooxygenase forms, were used as test agents. Their effects were compared to those of the directly alkylating agents N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) and N-ethyl-N nitrosourea (ENU). The cytotoxicity was evaluated by plating efficiency, the genotoxicity by the appearance of alkaline labile DNA sites. The nitrosamines had a cytotoxic and genotoxic effect on the differentiated HF1-4 cells, but had no effect on H5 cells. Aflatoxin B1 affected both cell lines, but was approximately 10-times more potent in the HF1-4 than in the H5 cells. In contrast to the nitrosamines and the mycotoxin, benzo[a]-pyrene exerted a stronger effect on the dedifferentiated cell line. Pretreatment of cultures with dexamethasone increased both the cytotoxicity and genotoxicity of the hepatotoxic agents. MNNG and ENU induced a similar degree of DNA-damage after short-term (2 h) exposure in the two cell lines. When cells were allowed to recover for 16 h HF1-4 cells, but not H5 cells, regained their full growth potential suggesting a marked capacity for the repair of MNNG- and ENU-induced lesions in the HF1-4 cells. The results indicate that continuous lines of mammalian cells may retain a considerable degree of organ-specific response to chemical carcinogens. Hepatoma cells of the type described above may be useful for screening the wide spectrum of chemicals which are potentially genotoxic in liver and in extrahepatic tissues and for analyzing their metabolic activation and mechanism of action. PMID- 6293735 TI - Epigenetic membrane effects of a possible tumor promoting type on cultured liver cells by the non-genotoxic organochlorine pesticides chlordane and heptachlor. AB - The organochlorine pesticides chlordane and heptachlor were evaluated for their genotoxicity and epigenetic membrane effects. Both compounds were non-genotoxic in the ARLHGPRT mutagenesis assay in which the genotoxic carcinogens 7, 12 dimethylbenz[a]anthracene and benzo[a]pyrene induced significant increases in mutant incidence. The pesticides both markedly inhibited intercellular communication between cultured liver cells, a property demonstrated by many tumor promoting agents, whereas benzo[a]pyrene did not produce this epigenetic effect. PMID- 6293737 TI - Chemical interactions with herpes simplex type 2 virus: enhancement of transformation by selected chemical carcinogens and pro-carcinogens. AB - Quantitative assays for the morphological transformation of 3T3 Swiss mouse cells by herpes simplex type 2 virus (HSV-2) were employed to examine the effect on cell transformation of chemical carcinogens and pro-carcinogens. The carcinogens tested were N-methyl-N'-nitro-N-nitrosoguanidine, quinacrine mustard, N nitrosomethyl urea, urethane, and benzene. The pro-carcinogens tested were N nitrosodimethylamine, 3-methylcholanthrene, benzo[a]pyrene, and p dimethylaminoazobenzene. Exposure of the cells to the chemical compound and to the virus resulted in enhancement of transformation when compared to that observed with chemical or virus alone. Enhancement of transformation occurred in cells treated with all of these compounds. In general, enhancement occurred regardless of whether the cells were pre-exposed to the carcinogen or pre infected with virus. These results are suggestive of combined herpes virus and chemical effects on cells resulting in increased risk of oncogenic transformation. PMID- 6293738 TI - Stimulation of slow action potentials in guinea pig papillary muscle cells by intracellular injection of cAMP, Gpp(NH)p, and cholera toxin. AB - To test the hypothesis that intracellular cAMP has a regulatory role in cardiac slow channel function, intracellular pressure injections of cAMP and adenylate cyclase activators, Gpp(NH)p and cholera toxin, were carried out. Guinea pig papillary muscles were depolarized to about -45 mV by superfusion with 22 mM K+ Tyrode's solution to inactivate the fast Na+ channels. Induction of slow action potentials or enhancement of ongoing slow action potentials was observed in about 70% of all cells in which a successful intracellular injection of the testing compounds was obtained. The slow AP is highly dependent on slow inward current and is known to be enhanced by catecholamines. The effect of the injected cyclic nucleotides and related compounds occurred within 3 minutes after starting the injection, whereas superfusion with these compounds (dibutyryl cAMP was used in place of cAMP) required 10-30 minutes to show an effect. This difference is attributed to the intracellular injection of the compound. The effect on stimulating slow action potentials persisted (greater than 5 minutes) after termination of the application of either Gpp(NH)p or cholera toxin, indicating the long-lasting nature of their action. The effect of the cAMP injections decayed within 1 minute. Intracellular injection of 5'-AMP was without effect. These results support the view that a causal relationship exists between intracellular cAMP level and slow channel function. Phosphorylation of a protein constituent of the slow channel by a cAMP-dependent protein kinase may be involved. PMID- 6293739 TI - Radioimmunoassay for 1,25-dihydroxycholecalciferol. PMID- 6293740 TI - An acth-secreting pituitary tumour arising in a patient with congenital adrenal hyperplasia. AB - The case reported is of a 46-year-old woman who had congenital adrenal hyperplasia due to a 21-hydroxylase deficiency, and in whom there was the development of an ACTH secreting pituitary tumour. The patient was untreated with glucocorticoids until the age of 32 years when she presented with infertility. She next presented with amenorrhoea at the age of 44 years when she was found to have an enlarged pituitary fossa. Despite treatment with bromocriptine and adequate doses of dexamethasone, the tumour enlarged and required operative treatment 1 year later. Before and after operation, plasma ACTH levels were between 300 and 400 ng/l, immunocytochemistry showed staining for ACTH and other structurally related pro-opiocortin peptides but for no other hormones, and the tumour secreted large amounts of ACTH in vitro. The report of this case is to our knowledge the first account of a feedback tumour in congenital adrenal hyperplasia and provides yet another reason why patients with this condition should be treated, and good control achieved. PMID- 6293741 TI - Direct radioimmunoassays of renin and renin substrate during converting-enzyme inhibition. AB - In 10 severely hypertensive patients, on a low sodium diet, converting enzyme inhibition increased plasma renin activity and decreased plasma renin substrate. The use of direct radioimmunoassays for both the enzyme and its substrate showed that the number of immunoreactive renin molecules increased from 11.3 +/- 4.9 to 31.7 +/- 25.3 pmol 1(-1) whereas the number of immunoreactive renin substrate molecules decreased from 1.04 +/- 0.35 to 0.74 +/- 0.16 mumol 1(-1). The direct radioimmunoassay for angiotensinogen gave higher values than the direct enzymatic assay, and during converting enzyme inhibition, the difference between both methods increased in proportion to the rise in circulating renin. It is concluded that the difference between the renin substrate radioimmunoassay, which measures angiotensinogen and des-angio I-angiotensinogen, and the renin substrate enzymatic assay which only measures "active" substrate, is an index of the increased consumption of renin substrate, in a situation where the fall in angiotensin II enhances renin release and decreases renin substrate release. PMID- 6293743 TI - Lymphocyte subsets in hypogammaglobulinaemia. PMID- 6293742 TI - Enzymes of purine metabolism in human peripheral lymphocyte subpopulations. AB - Ecto-5'nucleotidase (5'NT), adenosine deaminase (ADA), purine nucleoside phosphorylase (PNP) and deoxycytidine (CdR), deoxyguanosine (GdR), deoxyadenosine (AdR) and adenosine (AR) kinases have been measured in subpopulations of peripheral blood lymphocytes of eight healthy volunteers. The separation of B, T, T helper/inducer and T suppressor/cytotoxic cells was performed by means of density gradient centrifugation, E rosetting, passage through a nylon-wool column and antibody affinity chromatography utilising OKT8 and OKT4 monoclonal antibodies. ADA was significantly higher in T lymphocytes and 5'NT in B lymphocytes. Among T cell subpopulations, 5'NT activity was significantly higher (P less than 0 . 01) in T suppressor/cytotoxic (OKT8+) cells (32 . 9 units/10(6) cells than in T helper/inducer (OKT4+) cells (9 . 7 units/10(6) cells). Indeed, the 5'NT activity in T suppressor cells was similar to that in B cells. T helper cells tended, however, to have higher PNP and ADA activities than T suppressor cells but the differences were not statistically significant. No major differences were noted in kinase activities between any of the lymphocyte subpopulations. PMID- 6293744 TI - Enzyme patterns in normal lymphocyte subpopulations, lymphoid leukaemias and immunodeficiency syndromes. PMID- 6293745 TI - [Electrophysiological aspects of peripheral nerve lesions as a sequela of SMON]. PMID- 6293746 TI - [A study of blood nerve barrier in experimental allergic neuritis]. PMID- 6293747 TI - [An autopsy case of extramammary Paget's disease with an acute onset of neurological deficit]. PMID- 6293748 TI - Imaging of brain tumors and other lesions utilizing Tc-99m phosphates and Tc-99m pertechnetate. AB - The intensity of parenchymal brain lesions was compared using Tc-99m pertechnetate and Tc-99m phosphate. The following conclusions were made: 1. If the Tc-99m phosphate intensity is greater than the intensity of the Tc-99m pertechnetate scan, and the patient is evaluated within four weeks of ictus, the lesion is a CVA (P less than .001). 2. If the Tc-99m phosphate intensity is less than or equal to the intensity of the Tc-99m pertechnetate scan, and the patient is evaluated within four weeks of ictus, the parenchymal lesion is not a CVA (P less than .001). 3. If the evaluation takes place longer than six weeks after ictus, then no evaluation about the nature of the lesion can be made based upon uptake of Tc-99m phosphate and Tc-99m pertechnetate. PMID- 6293749 TI - Diagnosis of non-Hodgkin's lymphoma intracerebral mass lesions. Usefulness of Tc 99m pertechnetate and Ga-67 citrate brain scans. AB - This paper summarizes the clinical and diagnostic features of five reports of patients with intracerebral, non-Hodgkin's lymphoma. In three patients the brain lesion was the only evidence of lymphoma, while two patients also had concomitant systemic involvement. Four patients had diffuse histiocytic lymphoma and one had a mixed type of malignant lymphoma. In all patients, Tc-99m and Ga-67 brain scans disclosed discrete areas of increased radionuclide uptake consistent with a mass. In each case, brain blood perfusion studies were normal and brain computerized tomographic (CT) scans and cerebral angiograms produced variable nondiagnostic patterns. Craniotomies in four patients provided histologic confirmation of the non-Hodgkin's lymphoma in the areas of abnormality. The remaining patient had systemic histiocytic lymphoma with concomitant brain lesions that responded to irradiation. The combined use of the above noninvasive modalities in correlation with clinical findings may result in more accurate prebiopsy diagnoses of intracerebral lymphoma. PMID- 6293750 TI - Left ovarian vein. Unexpected renal scan finding. PMID- 6293751 TI - Incidence of a pyramidal lobe on thyroid scans. AB - Gamma camera pertechnetate and radioiodine thyroid scans were reviewed to determine the incidence of recognition of a pyramidal lobe. Ten to 17% of "normals" and of patients with various thyroid disease states had a pyramidal lobe on their scans. However, in patients with diffuse toxic goiter, 43% had a pyramidal lobe on the thyroid images. There appears to be a correlation between elevated thyroid function studies (likely in thyroid mass) and the incidence of a pyramidal lobe on thyroid scans in diffuse toxic goiter. PMID- 6293752 TI - Disease of the trophoblast. AB - Molar pregnancy should always be considered as a possible diagnosis in any pregnancy that does not conform to the normal. The practice of subjecting all pregnant women to ultrasound B-scan at their first visit in some centres is commendable, but unfortunately not possible in the areas of high incidence, owing to lack of facilities. Close, regular and meticulous follow-up with facilities for radio-immunoassay of HCG is a necessity after a mole has been treated. Early detection of lesions and individualized treatment of patients, with close monitoring of progress, are essential. Where necessary, enlightened use of drugs with adjuvant surgery and occasionally radiotherapy will give the optimum chance of recovery. To achieve complete eradication of the tumour, treatment may be required even after the HCG test appears to be negative. PMID- 6293754 TI - Variations in the immune response to Herpesvirus saimiri in squirrel and rhesus monkeys. AB - Humoral and cell-mediated immunity (CMI) to herpesvirus saimiri (HVS), an oncogenic lymphotropic herpesvirus, was studied in squirrel and rhesus monkeys. Natural antibody to HVS was found in five of six squirrel monkeys but there was no evidence of specific CMI directed against HVS. Rhesus monkeys did not show natural antibody or CMI against HVS antigens. Immunization with HVS, however, produced both antibody and specific CMI in the rhesus monkeys, but no CMI developed in the squirrel monkeys. These findings are important in the development of animal models for the treatment of tumors associated with lymphotropic herpesviruses. PMID- 6293753 TI - Sexually transmitted diseases in pregnancy. PMID- 6293755 TI - [An ELISA test for detection of maedi-visna antibodies. Comparative study with gel immunodiffusion and complement-fixation test]. AB - An indirect microELISA test was performed for detection of maedi-visna antibodies in ovine and caprine species. The antigen consisted in viral particles, highly purified by successive ultracentrifugations. By comparative testing of 934 sera in ELISA and gel immunodiffusion, we found a good correlation between these two tests, and moreover, ELISA revealed another 11.3% of positive samples. The precocity of this ELISA was shown by experimental infection of sheep with different strains of maedi-visna: positive sera were detected 7 weeks post infection, instead 4-5 months with gel immunodiffusion. The complement fixation test was compared with gel immunodiffusion and was found the less sensitive. This ELISA test appeared to be satisfactory, and may be used for early diagnosis of maedi-visna infection. PMID- 6293756 TI - Biochemical and immunological studies of angiotensin converting enzymes from human, bovine, dog, hog, rabbit, rat and sheep kidneys. AB - 1. Molecular and kinetic properties of angiotensin converting enzymes from seven different species' kidneys were similar concerning apparent molecular weight, heat sensitivity, Km value, optimum pH, activation by chloride ion, and inhibition by specific converting enzyme inhibitors (captopril and SA 446). 2. Rabbit antibody against pure human kidney enzyme cross-reacted partially with each animal kidney enzyme except for the rabbit enzyme. Antigenic determinants of animal enzymes were variable from species to species and differed from those of the human enzyme in extent and specificity. PMID- 6293757 TI - Distribution and properties of flavokinase in the developing chick embryo. AB - 1. Localization of flavokinase was studied in the nonembryonic portions of the chicken egg, in embryos at various stages of development, and in the adult hen. 2. Embryonic biosynthesis of flavin nomonucleotide (FMN) increases with increasing protein synthesis. 3. In the embryo, higher activity of flavokinase was observed in intestine, heart, liver, and yolk sac. 4. Lower activity was observed in allantoic membrane and fluid. 5. No significant activity was measured in yolk. 6. Both specific and total flavokinase activities in organs of the more aerobic and metabolically active adult chicken are considerably greater than those of embryo organs. 7. Flavokinase from embryonic liver was enriched by affinity chromatography by using N-10-flavinyl agarose. 8. The partially purified enzyme has properties generally similar to other animal flavokinases. PMID- 6293759 TI - Report of a clinical conference on a new implant material for ridge augmentation and preprosthetic preparation of the edentulous patient. PMID- 6293758 TI - Influence of hormone concentration and time factor on development of receptor memory in a unicellular (Tetrahymena) model system. AB - 1. Treatment of Tetrahymena pyriformis cells with diiodotyrosine (T2) gave rise to a considerable, concentration-dependent increase of the growth rate within the range of 10(-15) and 10(-9) M, but did not influence it at the level of 10(-18) M. 2. Re-exposure of the cells 1, 2 and 4 weeks later to the hormone concentrations originally used accounted for a marked increase of growth rate at all hormone levels tested, indicating that the extremely low concentration of 10( 18) M, which failed to stimulate growth on first exposure, did nevertheless give rise to hormonal imprinting, which caused the cells to "remember" the hormone, as judged from their increased responsiveness to it on re-exposure. 3. The degree of growth response was concentration-dependent on both first and second exposure: higher levels of treatment gave rise to firmer imprinting, and to greater response on re-exposure. 4. The length of exposure time proved to be more decisive than the level of treatment in respect of the development of hormonal imprinting. 5. Short-term exposures up to 60 min, although they stimulated cell growth by direct effect, gave rise to lasting inhibition of cellular response to re-exposure(s) rather than to hormonal imprinting. PMID- 6293760 TI - Review of clinical experiences with durapatite. PMID- 6293761 TI - A new implant material for ridge augmentation and preprosthetic preparation of the edentulous patient. Review of clinical experiences. PMID- 6293762 TI - Observations and reflections on durapatite. PMID- 6293763 TI - Impact of durapatite as a bone grafting material in oral and maxillofacial surgery. PMID- 6293764 TI - Catechol estrogens and thrombosis: differential effect of 2-hydroxyestradiol and estradiol on prostacyclin release. AB - High estrogen burden states, such as that seen in women who ingest oral contraceptive preparations, are associated with increased thrombosis. In other instances, however, estrogens decrease thrombogenicity. Variations in conversion to active metabolites could explain this paradox. Thus, we examined the influence of estradiol and a major metabolite, 2-hydroxyestradiol (catechol estrogen), on prostacyclin release from cultured human umbilical venous endothelium. In micromolar concentrations, both compounds inhibited prostacyclin (PGI2) release from bradykinin-, arachidonic acid-, and ionophore-stimulated, as well as unstimulated monolayers. However, when thrombin was used to stimulate release, estradiol enhanced, while 2-hydroxyestradiol inhibited, PGI2 formation. These sex steroids probably exert their influence at micromolar concentrations through a non-specific interaction with the cell membrane, and may be of importance in the predisposition to thrombosis suffered by those who use oral contraceptives. PMID- 6293765 TI - Immunosuppressive therapy in primary glomerulonephritis (pros). PMID- 6293766 TI - The effect of cyclophosphamide on the myocardial oxidase system of mitochondria. AB - The authors studied the effect of immunosuppressive doses of cyclophosphamide (5 mg/kg b. w.) on the activity of myocardial NADH, succinate and cytochrome oxidases in the mitochondria of rabbit myocardium on two experimental models: a) after a single dose of cyclophosphamide (acute model); b) after cyclophosphamide given daily for a period of 10 weeks (chronic model). A single dose of cyclophosphamide did not affect the myocardial oxidase systems of mitochondria, whereas its chronic administration significantly increased the succinate oxidase activity and decreased cytochrome oxidase activity. The drug did not interfere with NADH- oxidase activity. It is concluded that long-term administration of cytostatic and immunosuppressive doses of cyclophosphamide may result in severe myocardial lesions on subcellular level. This should be taken into account in clinical practice. PMID- 6293767 TI - Transforming genes and gene products of polyoma and SV40. AB - The small DNA-containing viruses, SV40 and polyoma, transform cells in vitro and induce tumors in vivo. For both viruses two genes required for transformation have been found. The genes required for transformation are also involved in productive infection. Although the two viruses are similar in their effects on cells, the organization of the transforming genes and gene products is different. The purpose of this review is to compare what is known about the biology and the biochemistry of the early regions of the two viruses. The genetic and biochemical studies defining the sequences important for transformation will be reviewed. Then, the products of the transforming genes, called T antigens, will be discussed in detail. There is a substantial body of descriptive information on those products, and studies on the function of the T antigens have also begun. PMID- 6293768 TI - Studies on sequence recognition by type II restriction and modification enzymes. AB - Type II DNA restriction and modification systems are ideally suited for analysis of mechanisms by which proteins specifically recognize unique DNA sequences. Each system is comprised of a unique DNA recognition site and two enzymes, which in those cases examined in detail, are comprised of distinct polypeptide chains. Thus, not only are the DNA substrates extremely well defined, but each system affords the opportunity to compare distinct proteins which interact with a common DNA sequence. This review will focus only on those Type II systems which have been examined in sufficient molecular detail to permit some insight into modes of specific DNA-protein interaction. PMID- 6293769 TI - Immunotoxicology of silica. PMID- 6293770 TI - Chondroid syringoma in a toe. AB - The second case of chondroid syringoma in a toe is presented. The diagnosis was confirmed microscopically, and histochemical studies suggested the presence of heavily sulfated stromal acid mucopolysaccharides. Glycogen was present only in some tumor cells. Keratin-containing tumor cells and nests were seen with polarization. This tumor should be considered in the differential diagnosis of painless, slow-growing, pedal tumors. PMID- 6293771 TI - Chromoblastomycosis treatment with ketoconazole. PMID- 6293772 TI - The intracellular acid-extractable (acid-soluble) amino acid pool in mammalian cells: 4 An hypothesis to explain the effects of preloading on the exchange of amino acids across the cell membrane. AB - Intracellular acid-extractable pools preloaded with amino acids before the cells were placed in lower concentrations of labelling amino acid showed a massive infiltration of the label at resuspension. In some the overshoot was rapid, in others it was slower developing, but in all cases the final level returned to the control value. Some amino acids can produce an overshoot in preloaded pools of others, whereas in certain combinations, e.g. alanine preloaded cells challenged with leucine, no discernible change in uptake kinetics was seen. Discharge of the preloaded pool is a necessary prerequisite for overshoot since cells placed in challenges of equal concentration to the preloading conditions do not exhibit this phenomenon. There is no energy requirement for the rapid phase of overshoot to occur since it follows similar kinetics at 2 degrees and 37 degrees C. The results are discussed in terms of exchange reactions occurring within the acid extractable pool which is composed of amino acids in a complexed form, and a hypothesis is offered to explain the phenomenon. This hypothesis is consistent with the cyclical perfusion model of Wheatley and Inglis (1980), and does not contravene Fick's law ans the second law of thermodynamics. PMID- 6293773 TI - Exogenous DNA transcription in cells with their native DNA inhibited. 4. Demonstration of specific antigens codified by exogenous DNA. AB - It is possible to obtain antisera to cancer-specific antigens of mouse Ehrlich ascites cancer (EAC) cells when chimera rabbits previously made unresponsive to immunological stimulation by normal mouse cells antigens are immunized with mouse cancer cells. Employing this specifically anti-cancerous immunoserum this work shows that EAC cells and TC-SV40 cells contain cross-reacting cancer-specific antigens. Exogenous DNA from EAC cells is demonstrated as being able to display its coded information when it is incorporated to endogenous TC-SV40 cell DNA whose activity has been previously inhibited by 5'-bromodeoxyuridine. PMID- 6293774 TI - [Ultrastructural study of mixed tumours]. PMID- 6293775 TI - Small cell lung cancer. Serial bronchofiberscopy and photographic documentation- the bridge sign. AB - To determine whether serial bronchofiberscopy provides better assessment of remission and relapse in chemotherapy and radiotherapy for small cell lung cancer, therapy was preceded by bronchofiberscopy with a new, high-resolution system for photographic documentation and either open-tube of bronchofiberscopic histocytologic diagnosis. Four-course cytostatic induction therapy was combined with prophylactic brain irradiation and irradiation of primary disease sites. Bronchofiberscopy was repeated after induction chemotherapy (in some cases combined with consolidating radiotherapy) and following complete remission (CR) if relapse was suspected. Sixty patients with limited-stage disease underwent a total of 152 bronchoscopies. Eight (16 percent) of those thought to be in CR by clinical, radiologic, and tumor-marker data were histocytologically tumor positive on endoscopy. Of ten patients thought later to have had relapse following CR, four were endoscopically tumor-positive. The bridge sign is described as a further endoscopic indicator of CR. Consecutive outpatient endoscopic examinations were well tolerated, without complications. In treatment of small cell lung cancer, routine serial bronchofiberscopy is feasible and useful. It substantiates definition of CR, lowering the risk of early relapse. By better identifying the partial responder, it contributes to a rational approach for subsequent management. Finally, it enables early endoscopic detection of local recurrence in selected cases. PMID- 6293776 TI - The roentgenographic staging of sarcoidosis. Historic and contemporary perspectives. AB - The awareness that sarcoidosis can involve intrathoracic lymph nodes and lung parenchyma in sequential fashion evolved during the first half of this century. This awareness resulted in a roentgenographic staging system that has relevance to the course and prognosis, pulmonary function changes, and the symptom of dyspnea. The following definition or roentgenographic staging is proposed as the simplest, most reproducible system based solely on the roentgenographic appearance, avoiding histopathologic and pathophysiologic inferences. Stage I: bilateral hilar lymphadenopathy; stage II: bilateral hilar lymphadenopathy plus parenchymal infiltration; and stage III: parenchymal infiltration without bilateral hilar lymphadenopathy. The early data concerning bronchoalveolar lavage, determination of serum angiotensin-converting enzyme, and gallium 67 citrate scanning appear to have no clear relationships to staging, suggesting that staging continues to have unique value to the clinician in the classification of sarcoidosis and in the delineation of its clinical course and prognosis. PMID- 6293777 TI - The brain's own morphine and cigarette smoking: the junkie in disguise? PMID- 6293778 TI - Effects of traditional Chinese herbs, toad tincture and adenosine 3',5' cAMP on Ehrlich ascites tumor cells in mice. PMID- 6293779 TI - Computed tomography in diagnosis of hepatocellular carcinoma. PMID- 6293780 TI - [Cystic ganglion of the common peroneal nerve]. PMID- 6293781 TI - [Complications caused by tourniquets during operations on the limbs under ischemic conditions]. PMID- 6293782 TI - [A rare cause of aorto-caval fistula: malignant fibrous retroperitoneal histiocytoma]. PMID- 6293783 TI - Overlapping deficiencies refine the map position of the sex-linked actin gene of Drosophila melanogaster. AB - A 3H-labelled actin-specific probe was hybridized to Drosophila melanogaster X chromosomes heterozygous for deficiencies in the 5C region. The results suggest that the sex-linked actin gene resides in the overlap region of Df (1) C149 and Df (1) N73 at 5C3-4. PMID- 6293784 TI - [Scanning electro microscopic photography of lesions induced by a single dose of ground quartz emulsion given intratracheally in rats]. PMID- 6293785 TI - [Reaction of pulmonary macrophages activated by BCG to silica. II. A study of the changes of the permeability of lysosomal membrane]. PMID- 6293786 TI - Chromosome sorting and DNA sequence localization. AB - This article focuses on current techniques and possible future developments in chromosome isolation and sorting, and DNA analysis of sorted chromosomes. The strategy of subchromosomal gene mapping by chromosome sorting is outlined and a list of cell lines containing translocated chromosomes is provided which may be used to map genes to a single chromosome with a standard fluorescence activated cell sorter. The usefulness of this and other gene mapping methods for localizing unique DNA sequences and characterizing recombinant DNA libraries constructed from sorted chromosomal DNA is also discussed. PMID- 6293787 TI - Analysis of ligand-receptor interactions with the fluorescence activated cell sorter. AB - The binding of a fluorescent peptide to human neutrophils is analyzed with a fluorescence activated cell sorter. We examine steady-state and kinetic features of the ligand-receptor interaction (in the presence of unbound ligand) and we show tht the number of receptors may be estimated without resorting to any external references for calibration. These methods are applicable to other fluorescent ligands with affinities greater than 10(8) M-1. PMID- 6293788 TI - Flow cytofluorometric study of lipoprotein interactions with cultured endothelial cells. AB - Accumulation of rhodamine B isothyocyanate-conjugated low density lipoproteins (R LDL) in cultured endothelial cells from human umbilical cord was studied with a fluorescence activated cell sorter. R-LDL uptake was blocked at 0 degrees C, inhibited by addition of excess of nonlabeled low density lipoprotein, high density lipoprotein-2 or high density lipoprotein-3. High density lipoprotein-2 was about twice as effective in inhibition of R-LDL uptake as high density lipoprotein-3. Endothelial cells that formed a contact-inhibited monolayer lost the ability to incorporate R-LDL via a receptor-mediated pathway. Using R-LDL, it was possible to distinguish cells with different levels of R-LDL incorporation. PMID- 6293789 TI - A strategy for prevention of cancer of the large bowel. AB - The etiology of cancer of the large bowel is in large part environmental. Both epidemiologic and experimental studies suggest that the environmental factor is dietary, specifically associated with a diet high in fat and low in fiber. Prevention is possible by altering these dietary factors, but the degree of change needed to accomplish it would be too drastic to be acceptable. However, the inclusion of certain additives in a diet only slightly modified with respect to the fat-fiber content may circumvent this problem. We have evidence that the addition of small amounts of representatives of three classes of cancer inhibitors (retinoids, plant steroids, and selenite) reduces intestinal cancer formation in rats by 50 per cent. This suggests that a diet with modest changes in the fat-fiber content plus the addition of certain inhibitors may be effective in reducing the incidence of cancer of the large bowel. PMID- 6293790 TI - Review of sigmoid volvulus. Clinical patterns and pathogenesis. PMID- 6293791 TI - Increased insulin receptor binding to monocytes from insulin-dependent diabetic patients after a low-fat, high-starch, high-fiber diet. PMID- 6293792 TI - [Isolated gastrointestinal tract synthesizes ACTH and possibly thyroid hormones and enkephalins]. PMID- 6293793 TI - Amitriptyline-related peripheral neuropathy relieved during pyridoxine hydrochloride administration. AB - Tricyclic antidepressants rarely cause peripheral neuropathy. In fact, this class of drugs has been used to control the symptoms of pain and paresthesia that accompany peripheral neuropathy. We report peripheral paresthesias that occurred in a 39-year-old female during five years of amitriptyline administration. The patient's symptoms were relieved by oral pyridoxine hydrochloride, associated with elevated plasma pyridoxal phosphate. PMID- 6293794 TI - [Intestinal protozoal infestation in persons with occupational sewage contact]. AB - Intestinal protozoal infestation and antibodies against Entamoeba histolytica were determined in 614 employees of the Hamburg sewage works and compared with control groups without occupational sewage contact. The protozoal infection rate was significantly higher in sewer-men than in control persons. There was a positive correlation between the duration of exposure to sewage and the frequency of intestinal protozoal infestation. The risk of infection by sewage was higher than by a short journey to tropical or subtropical countries. Immunodiagnostic results showed no significant differences of invasive activity of E. histolytica between both investigated groups. On the basis of parasitological results classification of amoebiasis and giardiasis as occupational diseases in exposed persons is recommended. PMID- 6293795 TI - Adolescent hypophosphataemic rickets. PMID- 6293796 TI - [Naloxone-resistant EEG slowing induced by the synthetic opioid peptide FK 33-824 in the 4th cerebral ventricle of the dog]. AB - In order to determine the importance of opioid peptides in the central control of wakefulness, the synthetic analogue of Met-Enkephalin, FK 33-824 (d-Ala-2-Phe-Met (O)-ol-Enkephalin) which is more resistant to enzymatic degradation, was perfused in increasing concentrations (20, 100, 200, and 400 micgrograms/ml) through the fourth cerebral ventricle of the conscious dog. In the EEG (Power-spectral density in continuous acquisition) high concentrations (200--400 micgrograms/ml) induce slowing (theta--delta) with overlying beta-activity. This was reflected in the animals behaviour resulting in a sleep-like state which in spite of the administration of high doses of the antagonist Naloxone (100 micrograms/kg i.v.) sustained for 12 hours. A simultaneously dose-related respiratory depression (drop in arterial pO2 and an increase in arterial pCO2) was reversed by Naloxone. It is concluded that the opioid peptide FK 33-824 induces EEG-slowing which is mediated by a subpopulation of opiate-binding sites, possibly the kappa receptors. These receptors are different from binding sites mediating respiratory depression (micro-receptors) as they interact with the opiate-antagonist Naloxone. PMID- 6293797 TI - Structural and functional organization of some pituitary protein hormones. AB - Some recent observations on the interrelation between structural and functional organization of some anterior pituitary hormones (corticotropin, beta-lipotropin, prolactin and somatotropin) made in this Institute are reviewed. The primary structure and interspecies differences of these hormones are discussed and a mathematical method of comparative analysis of the homologous amino acid sequences, which has been previously developed, is presented. A hypothesis has been raised that some sequences of the amino acid chains are closely related to the nature of biological effect of the appropriate protein hormone, while some other parts of the molecular are responsible predominantly for a contact with a specific receptor and several experimental data are presented which seem to be in agreement with such a view. This may be also supported by the observations that the lipolytic effect of somatotropin may be blocked by proteinase inhibitors (trasylol and methylamine) which shows that such an action presumably takes place only after a cleavage of its molecule by proteolytic enzymes into smaller fragments. PMID- 6293798 TI - Three types of desensitization of metabolic responses to thyrotropin in thyroid tissue: a review. AB - The evidence is reviewed that a desensitization to thyrotropin is of three distinct types. Type 1 is proximal to the production of cyclic AMP and is manifested by a fall of cyclic AMP concentration in the thyroid tissue despite the continued presence of TSH or phosphodiesterase inhibitors in the medium. Type 2 is distal to this event and involves the metabolic effects of TSH mediated by cyclic AMP such as colloid droplet formation, iodide organification and, in dog thyroid, glucose oxidation. All these effects may be due to a reduced amount of cyclic AMP which is not capable of stimulation the protein kinase. They may be prevented by a pre-exposure to similar concentrations of TSH and cannot be overcome by other stimulators acting through the same adenylate cyclase-cyclic AMP pathway. Type 3 involves the processes not regulated by cyclic AMP. Thus, a desensitization of thyroid tissue by pre-exposure to TSH results in a reduced response of phospholipid turnover, a process not mediated by cyclic AMP, either to homologous (TSH) or heterologous (acetylcholine) stimulation. PMID- 6293799 TI - alpha-Melanocyte-stimulating hormone-like peptides in the intermediate lobe of the rat pituitary gland: characterization of content and release in vitro. AB - Reverse phase high performance liquid chromatography (HPLC) followed by RIA of the chromatographic fractions was used to separate and quantify, respectively, the alpha MSH-like peptides stored in the intermediate lobe (IL) of the rat pituitary gland and released from IL cells in vitro. Immunoreactive material eluting with the same HPLC retention time as N,O-diacetyl alpha MSH accounted for approximately 80% of the total immunoreactive alpha MSH (IR-alpha MSH) in either the neurointermediate lobe or dispersed IL cells. The remainder of the IR-alpha MSH coeluted with either synthetic desacetyl alpha MSH or alpha MSH. Furthermore, the predominant alpha MSH-like compound released in vitro from dispersed IL cells eluted from the HPLC column with the same retention time as synthetic N,O diacetyl alpha MSH. Treatment of dispersed IL cells with drugs known to enhance (l-isoproterenol or A 23187) or to inhibit (apomorphine or lisuride) the release of IR-alpha MSH revealed that N,O-diacetyl alpha MSH was the primary form released. Finally, an evaluation of the stability of the alpha MSH-like peptides indicated that N,O-diacetyl alpha MSH was readily converted to alpha MSH in the presence of 0.1 N hydrochloric acid. PMID- 6293800 TI - Adenine-derived purines increase adenosine triphosphate (ATP) levels in the luteal cell: evidence that cell levels of ATP may limit the stimulation of adenosine 3',5'-monophosphate accumulation by luteinizing hormone. PMID- 6293801 TI - Specificity of the alteration in aldosterone biosynthesis in the spontaneously hypertensive rat. PMID- 6293802 TI - Effects of age and adrenocorticotropin on microsomal enzymes in guinea pig adrenal inner and outer cortices. AB - Guinea pig adrenals possess a distinctive zona reticularis which increases in volume as the animal matures, coming to occupy over 50% of the gland in mature 800-g animals. The adrenocortical cells of this zone possess abundant smooth surfaced endoplasmic reticulum. In this paper we demonstrate that microsomes obtained from guinea pig inner cortices of both young and mature animals, consisting predominantly of zona reticularis tissue, possess higher activities of 21-hydroxylase, an enzyme that hydroxylates steroids, and ethylmorphine demethylase, a drug-metabolizing enzyme, as well as higher contents of cytochromes b5 and P450, than microsomes derived from outer cortices, which contain predominantly zona fasciculata tissue. In addition, we show that while the inner cortex contains 90% of the total adrenal activity for ethylmorphine demethylase, ACTH suppresses this activity by 55% and decreases activity in the outer cortex to negligible levels. ACTH does not affect 21-hydroxylase activity or cytochrome b5 or P450 content in either cortical region. These observations suggest that differential regulation of microsomal functions for steroid hydroxylation and drug metabolism occurs largely in the zona reticularis. Furthermore, they suggest that other activities for xenobiotic metabolism, previously reported as a property of whole guinea pig adrenals, may reside in the inner cortex and be unique functions of the abundant smooth endoplasmic reticulum of the zona reticularis cells. PMID- 6293803 TI - Stress-induced secretion of adrenocorticotropin, corticosterone, and prolactin in experimentally and genetically hypertensive rats. AB - Plasma levels of immunoreactive ACTH, corticosterone (CS), and PRL in two-kidney, one clip (2K1C) hypertensive SABRA, hypertension-prone (SBH), hypertension resistant (SBN), and normotensive SABRA rats were compared under both quiescent conditions and after acute (2 min) cold water stress. Serum levels of CS were higher in 2K1C hypertensive compared with normotensive SABRA rats under both quiescent and stressful conditions. Circulating levels of ACTH and PRL were similar in both groups under quiescent conditions. Resting circulating levels of CS were higher in the SBH rats compared with SABRA or SBN rats. Serum PRL levels were similar in SBH and SABRA rats under both quiescent and stressful conditions. Resting PRL levels in the SBN rats were lower compared with the SABRA rats. Resting serum levels of ACTH and CS in the SBN rats were similar to those found in the SABRA rats. After stress exposure serum ACTH and CS levels were elevated in all groups. Serum PRL levels in SBN rats were not affected by stress, unlike the marked elevation seen in the other groups. Our study demonstrates increased secretion of CS in both 2K1C hypertensive and SBH rats under quiescent conditions. Both 2K1C hypertensive and SBH rats have normal hormonal capacity to respond to stress. SBN rats exhibited reduced PRL secretion under both quiescent and stressful conditions. It is suggested that abnormal activity of the hypothalamic-pituitary-adrenal system may play a role in the pathogenesis of 2K1C and genetic hypertension as well as in resistance to hypertension. PMID- 6293804 TI - Testosterone stimulation of progesterone synthesis by gonadotropin-desensitized corpora lutea. PMID- 6293805 TI - Insulin-induced hypoglycemia in conscious dogs: effect of maintaining carotid arterial glucose levels on the adrenocorticotropin, epinephrine, and vasopressin responses. PMID- 6293806 TI - Adrenalectomy inhibits noradrenergic, adrenergic, and vasopressor responses to angiotensin II in the Pekin duck (Anas platyrhynchos). AB - The iv injection of Asp1, Val5-angiotensin II(AII) was followed by increases in arterial plasma norepinephrine (NE) and epinephrine (E) concentrations and elevation of arterial blood pressure in the anesthetized Pekin duck. AII injection did not affect the concentration of unconjugated dopamine in arterial plasma. Adrenalectomy inhibited the increases in plasma NE and E concentrations elicited by AII. Pressor responses to AII also were diminished after adrenalectomy, whereas pressor responses to exogenous NE and tyramine were not measurably affected. The alpha-adrenergic blocking drug phenoxybenzamine diminished the pressor effect of AII in both shamoperated and adrenalectomized ducks. The blockade of alpha-adrenergic receptors by phenoxybenzamine was incomplete, since pressor responses to exogenous NE were lessened by not abolished. We conclude from these experiments that AII increases arterial blood pressure in the Pekin duck by mobilizing NE and E from both adrenal and extraadrenal stores. It remains unknown whether the residual pressor response to AII in phenoxybenzamine-treated, adrenalectomized ducks involves an alpha adrenergic mechanism. PMID- 6293807 TI - Thyroid hormone stimulation of phosphorylation and dephosphorylation of rat liver cytosolic proteins. PMID- 6293808 TI - Responses of cortisol secretion to repeated hemorrhage in the anesthetized dog. AB - We used two sequential 7.5 ml/kg hemorrhages, spaced 24 h apart, in the chronically prepared, pentobarbital-anesthetized dog to study the effects of repeated stimuli on the adrenocortical system. Adrenal secretion of cortisol, peripheral cortisol, and ACTH were measured. All three variables increased after an initial 7.5 ml/kg hemorrhage. When the hemorrhage was repeated 24 h later, the secretory response of cortisol began more rapidly (by 4 min), reached a higher peak, and was more prolonged than the response on day 1. ACTH rose to significantly higher values than on day 1, but only after 8 min. There was no differences in cardiovascular variables after hemorrhage on the 2 days. A repeated 3.75 ml/kg hemorrhage did not lead to a potentiated response. These results confirm reports that after physiological stimulation, changes occur in the pituitary-adrenal system that may lead to a potentiated response to later stimuli. The mechanism of these changes is unknown, but our results suggest that both an increase in circulating ACTH and a change in adrenal sensitivity to ACTH may be involved. PMID- 6293809 TI - Rat kidney cells in primary culture: interaction of salmon calcitonin with receptor sites. PMID- 6293810 TI - The effect of serotonin agonist 1-(trifluoromethylphenyl)-piperazine on corticotropin releasing factor and arginine vasopressin in rat hypothalamic nuclei. AB - Effects of 1-(m-trifluoromethylphenyl)-piperazine, a serotonin agonist, were examined on rat plasma levels of adrenocorticotropin (ACTH) and arginine vasopressin (AVP), and on hypothalamic contents of corticotropin releasing factor (CRF) and AVP, to investigate the role of brain serotonin in ACTH regulation. Both plasma ACTH and AVP levels increased markedly 30 min after injection of the compound and were still elevated at 80 min. CRF and AVP contents in the median eminence decreased 30 min after injection but returned to the basal levels by 80 min. The AVP content in the supraoptic nucleus was elevated 80 min after injection. The CRF and aVP content did not significantly change in the paraventricular, suprachiasmatic and arcuate nuclei. Serotonin or 1-(m trifluoromethylphenyl)-piperazine did not stimulate the release of ACTH in pituitary cell cultures. These results suggest that both CRF and AVP were secreted into the portal vessels by 1-(m-trifluoromethylphenyl)-piperazine to release ACTH from the anterior pituitary and that both the ACTH and AVP release were stimulated via the brain serotonergic mechanism. PMID- 6293811 TI - In vitro activity of N-formimidoyl-thienamycin in comparison to that of moxalactam and cefotaxime against gentamicin-resistant gram-negative bacteria. AB - The inhibitory and bactericidal activity of N-formimidoyl-thienamycin in vitro against 131 clinical isolates selected for their gentamicin resistance was compared with that of cefotaxime and moxalactam. All strains were inhibited by N formimidoyl-thienamycin concentrations within a range of 0.12-4 mg/l. N formimidoyl-thienamycin was less active than cefotaxime and moxalactam against Escherichia coli and Klebsiella spp., and more active than all other antibiotics tested against Serratia spp., Enterobacter cloacae, Pseudomonas aeruginosa and Acinetobacter spp. In contrast to the other antibiotics N-formimidoyl-thienamycin showed a narrow margin of difference between minimal inhibitory and minimal bactericidal concentrations. N-formimidoyl-thienamycin is a promising antibiotic for the treatment of hospital infections with multi-resistant organisms. PMID- 6293812 TI - Effect of subinhibitory antibiotic concentrations on the phagocytosis of Staphylococcus aureus. AB - The effect of subinhibitory antibiotic concentrations on the phagocytosis of Staphylococcus aureus was studied by pretreating 3H-thymidine labelled bacteria with one-third the minimal inhibitory concentration of clindamycin, doxycyclin, cefotiam, vancomycin, piperacillin and penicillin G, respectively. Pretreatment with clindamycin and doxycyclin resulted in enhanced uptake of the bacteria by polymorphonuclear leukocytes compared to the untreated control. The augmented phagocytosis was still observed at 1/32 the MIC of clindamycin and 1/64 the MIC of doxycyclin, and when the serum was diluted to a concentration of 1%. Pretreatment of the bacteria with penicillin, cefotiam, piperacillin and vancomycin had no effect on phagocytosis. Inhibitors of bacterial protein synthesis induce alterations of Staphylococcus aureus leading to increased phagocytosis, whereas antibiotics acting on cell wall synthesis are without effect. PMID- 6293813 TI - In vitro activity of ceftazidime, cefotaxime and piperacillin against multi resistant gram-negative bacteria tested with a modified agar dilution method. AB - The minimum inhibitory concentrations of ceftazidime, cefotaxime and piperacillin were determined for 870 multi-resistant gram-negative bacterial strains by means of a modified agar dilution technique using dishes with 25 compartments. This technique facilitates the reading of the MICs, reduces the risk of inoculation errors, and eliminates interference of interaction of bacterial strains. Ceftazidime and cefotaxime inhibited 90% of the Enterobacteriaceae strains at concentrations of 3.12 mg/l and 1.56 mg/l respectively. Ceftazidime and piperacillin inhibited 90% of the Pseudomonas aeruginosa strains at concentrations of 6.25 mg/l and 12.5 mg/l respectively. Mucoid Pseudomonas aeruginosa strains were more susceptible to the antibiotics tested, especially piperacillin, than non-mucoid strains. PMID- 6293814 TI - Adenosine diphosphate ribosylation of chicken-erythrocyte histones H1, H5 and high-mobility-group proteins by purified calf-thymus poly(adenosinediphosphate ribose) polymerase. AB - Poly(ADP-ribosylation) of histones H1, H5 and non-histone chromosomal high mobility-group proteins HMG 1, 2, 14 and 17 from chicken erythrocytes by purified calf thymus poly(ADP-ribose) polymerase was studied using acid/urea/Triton gel electrophoresis and autoradiography. With histone H1, besides ADP-ribosylated H1 supporting short chains of polymer, the appearance of H1 'dimer' was observed and this reaction was dependent on NAD concentration and incubation time. In addition, highly modified and/or aggregated species of histone H1 were observed. Histone H5 was slightly ADP-ribosylated at low NAD concentrations. At higher NAD concentrations or after longer incubations the formation of H5 'dimer' and of more modified forms of H5 could be observed. HMG 1 and HMG 2 were found to be ADP ribosylated, the reaction being dependent on NAD concentration and time. Here again some discrete intermediates appeared. HMG 14 and HMG 17 were only slightly ADP-ribosylated under our experimental conditions. These results indicate that the purified DNA-independent poly(ADP-ribose) polymerase can catalyse the formation of H1 'dimer' as in nuclei and nucleosomes and that H5 and HMG proteins can also be ADP-ribosylated and produce well-defined higher complexes. These modifications of nuclear proteins may provide a means of localized conformational changes of the chromatin structure in vivo. PMID- 6293815 TI - Phosphorylation of the type-II regulatory subunit of cyclic-AMP-dependent protein kinase by glycogen synthase kinase 3 and glycogen synthase kinase 5. PMID- 6293816 TI - Non-equilibrium thermodynamic assessment of redox-driven H+ pumps in mitochondria. AB - Isolated mitochondria suspended in an aerobic medium with 3-hydroxybutyrate or succinate serving as electron donor attain a stationary state with vanishing net flow of H+ ions (state 4). Adding valinomycin to such a suspension in the presence of various concentrations of K+ ions and a weak acid system such as acetate or phosphate creates new stationary states for the mitochondria which are characterized by a constant influx of K+ ions, while the net flow of H+ ions again vanishes due to the recycling of these ions by the weak acid system. Sufficiently low concentrations of K+ ions (less than 4 mM) cause these stationary states to last long enough for a separation of the mitochondria by centrifugation. The difference in electrochemical potential for H+ ions can then be determined by means of the partitioning of radioactively labelled markers. Suitable procedures to correct for binding of the markers are described. It is found that, for a constant affinity of the electron in the suspending medium, electron flow and the flow of K+ ions, which indicates the flow of pumped H+ ions, are linearly dependent on the electrochemical potential difference of H+ ions. The phenomenological coefficients obtained from these correlations are discussed with respect to the contributions of additive constants in the linear relations. It is found that, under the present experimental condition, such constants most likely vanish thus yielding symmetric flow-force relations. It is concluded that the redox-driven H+ pumps are not tightly coupled due to molecular slipping in the pumps and that the molecular stoichiometry is 2 H+ ions/electron for coupling site I and 4 H+ ions/electron for coupling sites II and III together. PMID- 6293817 TI - A shuttle mechanism for DNA-protein interactions. The regulation of poly(ADP ribose) polymerase. AB - Previously it had been shown that poly(ADP-ribose) polymerase requires DNA for its activity and that this enzyme is auto-poly(ADP-ribosyl)ated. The studies reported here indicate that this self-modification inhibits the enzyme and decreases its affinity for DNA, as shown by sucrose gradient density centrifugation. The coupling of poly(ADP-ribose) polymerase with poly(ADP-ribose) glycohydrolase reactivates the polymerase by degrading poly(ADP-ribose) and restoring the polymerase-DNA complex. The assay of polymerase in the presence of glyco-hydrolase was made possible by use of a double-label assay involving release of 14C-labelled nicotinamide and the incorporation of 3H-labelled ADP ribose from NAD+. These results provide the basis for a shuttle mechanism in which the polymerase can be moved on and off DNA by the action of these two enzymes. Mg2+ and histone H1 appear to activate the polymerase by increasing the affinity of the polymerase for DNA. PMID- 6293818 TI - A hypothesis for the role of dithiol-disulfide interchange in solute transport and energy-transducing processes. AB - We have recently shown that the physical mechanism for delta approximately mu H+ driven changes in the Km for three different transport systems is an oxidation reduction reaction involving a dithiol-disulfide interconversion [Robillard, G.T. and Konings, W.N. (1981) Biochemistry, 20, 5025-5032; Konings, W.N. and Robillard, G.T. (1982) Proc. Natl Acad. Sci. USA, in the press]. Based on the similarities between the data from these three systems and published data from other systems, we now propose that dithiol-disulfide interchange may play a general role in membrane-related processes such as transport, energy transduction and hormone-receptor interactions. We propose that the affinities of the substrate-binding sites are regulated by a dithiol and a disulfide situated at different depths in the membrane. In addition we propose that the oxidation states of these two redox centers are coupled by dithiol-disulfide interchange such that, when one is oxidized, the other is reduced. Since a transmembrane electrical potential, delta psi, or a pH gradient, delta pH, can alter the redox state, it can change the affinity of the substrate-binding sites. The delta approximately mu H+-induced changes in affinity are sufficient to drive active transport (symport or antiport) and energy-transducing processes. A similar mechanism can be applied to transport systems driven by phosphorylated enzyme intermediates instead of delta approximately mu H+. Changes of the redox potential in a given compartment during metabolism could also control the affinity of ligand binding even in the absence of a delta approximately mu H+. The ligand-binding affinities of facilitated diffusion transport systems and receptor proteins may be regulated in this manner. PMID- 6293819 TI - Phosphorylation and inactivation of yeast fructose-bisphosphatase in vivo by glucose and by proton ionophores. A possible role for cAMP. AB - Addition of glucose to yeast cells causes a phosphorylation and an inactivation of the gluconeogenic enzyme fructose-bisphosphatase [Mazon, M.J., Gancedo, J.M., and Gancedo, C. (1982) J. Biol. Chem. 257, 1128-1130]. We report here that the addition of the proton ionophores 2,4-dinitrophenol and carbonylcyanide m chlorophenylhydrazone to yeast cells produces the same effect as that of glucose. Both glucose and ionophores produced: (a) phosphorylation and inactivation of fructose-bisphosphatase, (b) an immediate rise in the intracellular concentration of cAMP, (c) an instant inhibition of the transport of amino acids driven by the membrane potential. It is proposed that the effect of glucose on fructose bisphosphatase involves as a first step the depolarization of the plasma membrane resulting in an increase of the intracellular concentration of cAMP. This in turn would stimulate phosphorylation of fructose-bisphosphatase. PMID- 6293820 TI - A pulse-radiolysis study of cytochrome c3. Kinetics of the reduction of cytochrome c3 by methyl viologen radicals and the characterisation of the redox properties of cytochrome c3 from Desulfovibrio vulgaris (Hildenborough). AB - 1. Pulse-radiolysis experiments were performed in the presence of methyl viologen and cytochrome c3. After the pulse, methyl viologen radicals are formed and the kinetics of these radicals with cytochrome c3 are studied, The reaction between cytochrome c3 and methyl viologen radicals (MV+) is diffusion controlled. The ionic strength dependence and the pH-dependence of this reaction were studied. From the ionic strength dependence (at pH 7.8) we found that the net charge of the fully oxidized cytochrome c3 molecule was Z = + 4.7 +/- 0.7. 2. After the pulse an equilibrium is reached for the reaction of MV+ with cytochrome c3. From this equilibrium an apparent midpoint potential can be obtained. The apparent midpoint potential of this multihaem molecule was found to depend on the degree of reduction, alpha. With the help of the Nernst equation an empirical equation is obtained to describe this dependence of the midpoint potential: E0 = - 0.250 - 0.088 alpha (in V). 3. An estimation is made of the energy of interaction between the haems due to electrostatic interactions (delta epsilon less than 32 mV) and due to ionic strength effects (- 12 mV less than delta epsilon less than 26 mV). The results suggest that the redox properties of the individual haems in the cytochrome c3 molecule are dependent on the degree of reduction of the other haems in the molecule. 4. The reaction of cytochrome c3 with MV+ or with ethanol radicals (EtOH) has been compared with the reactions of horse-heart cytochrome c and of metmyoglobin with the same radicals. The reaction of MV+ or EtOH with horse-heart cytochrome c is found to be diffusion controlled; the reactions with metmyoglobin on the other hand are most probably controlled by an activation energy. PMID- 6293821 TI - Studies on the mechanism of action of triphenyltin on proton conduction by the H+ ATPase of mitochondria. AB - A study is presented of the action of triphenyltin on the kinetics of the anaerobic relaxation of the proton gradient set up by respiration in various type of 'inside-out' inner membrane vesicles obtained by exposure of beef-heart mitochondria to ultrasonic energy. Triphenyltin is shown to act as a powerful inhibitor of the proton conductivity of the H+-ATPase. The inhibition persists after removal of the ATPase protein inhibitor, F1 and the oligomycin-sensitivity conferral protein (OSCP) from the particles. The inhibitory effect of triphenyltin is exerted, as in the case of oligomycin and N,N' dicyclohexylcarbodiimide, on the F0 moiety of the ATPase complex. Comparison of the characteristics of the effect of triphenyltin on proton translocation in chloride and nitrate media shows that the inhibition of passive proton conductivity studied here is unrelated to the hydroxide/anion exchange induced by the organotin. Lack of additivity of the inhibition of H+ conduction by triphenyltin with that exerted by oligomycin and N,N'-dicyclohexylcarbodiimide and the kinetic pattern of the effect of triphenyltin show that the mechanism of action of the organotin is different from that of the other two inhibitors. The relevance of the results obtained with respect to the subunit location and chemical nature of the reaction site of triphenyltin in the H+-ATPase complex is discussed. PMID- 6293822 TI - 1H-NMR studies on nucleotide binding to the sarcoplasmic reticulum Ca2+ ATPase. Determination of the conformations of bound nucleotides by the measurement of proton-proton transferred nuclear Overhauser enhancements. AB - The glycosidic bond torsion angles and the conformations of the ribose of Mg2+ATP, Mg2+ADP and Mg2+AdoPP[NH]P (magnesium adenosine 5'-[beta, gamma imido]triphosphate) bound to Ca2+ATPase, both native and modified with fluorescein isothiocyanate (FITC), in intact sarcoplasmic reticulum have been determined by the measurement of proton-proton transferred nuclear Overhauser enhancements by 1H-NMR spectroscopy. This method shows clearly the existence of a low-affinity ATP binding site after modification of the high-affinity site with FITC. For all three nucleotides bound to both the high-affinity (catalytic) site and the low-affinity site, we find that the conformation about the glycosidic bond is anti, the conformation of the ribose 3'-endo of the N type and the conformation about the ribose C4'-C5' bond either gauche-trans or trans-gauche. The values for the glycosidic bond torsion angles chi (O4'-C1'-N9-C4) for Mg2+ATP, Mg2+ADP and Mg2+AdoPP[NH]P bound to the low-affinity site of FITC modified Ca2+ATPase are approximately equal to 270 degrees, approximately equal to 260 degrees and approximately equal to 240 degrees respectively. In the case of the nucleotides bound to the high-affinity (catalytic) site of native Ca2+ATPase, chi lies in the range 240-280 degrees. PMID- 6293823 TI - Induction of lipogenic enzymes in primary cultures of rat hepatocytes. Relationship between lipogenesis and carbohydrate metabolism. AB - Using primary cultures of adult rat hepatocytes, the regulation of the following lipogenic enzymes was studied: glucose-6-phosphate dehydrogenase, malic enzyme, ATP-citrate lyase, acetyl-CoA carboxylase, fatty acid synthetase, and stearoyl CoA desaturase. The addition to the culture medium of either insulin or triiodothyronine produced a 2-3-fold increase in each of the individual enzyme activities whereas glucagon slightly decreased enzyme activities. The addition to the medium of 8-bromoguanosine 3,'5'-monophosphate had no effect on any of the enzyme activities unless glucose was also added to the culture medium. Glucose addition alone to the culture medium was without any effect; however, glucose enhanced the stimulation of enzyme activity due to insulin. The addition of fructose or glycerol, even in the absence of insulin, increased the activities of each of the enzymes studied 2-3-fold. The increases in enzyme activity brought about by insulin or fructose were apparently the result of de novo enzyme synthesis, as indicated by the observation that the increases were not noted in the presence of cordycepin or cycloheximide. Immunoprecipitation of ATP-citrate lyase from hepatocytes pulse-labeled with [3H]leucine indicated that the induction of this enzyme in response to the addition of fructose or glycerol to the culture medium was the result of an increase in the rate of synthesis of the enzyme. These results indicate that the activity and synthesis of individual enzymes involved in lipogenesis are increased in response to the metabolism of carbohydrate independently in part from hormonal effects. PMID- 6293824 TI - Induction of 2H+/Me2+ exchange in rat-liver mitochondria. AB - The time dependency of CA2+ efflux from Ca2+-loaded rat liver mitochondria has been investigated. The rate of ruthenium-red-insensitive Ca2+ efflux is continuously increased during the retention as a result of induction of an electroneutral H+ Ca2+ exchange system. The activation of the Ca2+ efflux pathway takes place under the constant value of the membrane potential and is accompanied by oxidation of mitochondrial pyridine nucleotides. It has also been found that the ruthenium-red-insensitive H+/Sr2+ exchange occurs in mitochondria during Sr2+ induced oscillation of ion fluxes. The rate of H+/Sr2+ exchange is variable and depends on the stage of the oscillatory cycle. PMID- 6293825 TI - Dual role of hexose-1-phosphate uridylyltransferase in galactosamine metabolism. PMID- 6293826 TI - Regulation of synthesis of catalases and iso-1-cytochrome c in Saccharomyces cerevisiae by glucose, oxygen and heme. AB - The regulation of the hemoproteins catalase T, catalase A and iso-1-cytochrome c was studied in the yeast Saccharomyces cerevisiae. Levels of catalase T and catalase A mRNAs are low or undetectable in anaerobic and heme-deficient cells, and in wild type strains grown on high glucose concentrations. Regulatory mutants (cgr4 and cas1), which have previously been shown to have high catalase T activity when grown in the absence of oxygen or on high glucose concentrations, have high levels of catalase T mRNA when grown under glucose repression conditions. Whereas no catalase T mRNA could be detected in a heme-deficient (ole3) single mutant, double mutants (ole3 cgr4) and (ole3 cas1) contain mature catalase T mRNA. Catalase T and A mRNAs are accumulated rapidly during adaptation of anaerobic cells to oxygen. Anaerobic and heme-deficient cells lack or have extremely low levels of iso-1-cytochrome c mRNA, which, like catalase mRNAs, is accumulated rapidly during oxygen adaptation. The results obtained demonstrate that glucose, oxygen and heme regulate the synthesis of the hemoproteins studied by controlling mRNA levels. In addition, posttranscriptional, probably translational control has to be postulated at least in the case of catalases, to explain the results obtained. PMID- 6293827 TI - The participation of poly(ADP-ribosyl)ated histone H1 in oligonucleosomal condensation. AB - The chromatin-associated enzyme poly(ADP-Rib) polymerase causes an NAD-dependent crosslinking of modified oligonucleosomes, as demonstrated by electrophoretic and sedimentation analysis [Butt, T. R. and Smulson, M. (1980) Biochemistry, 19, 5235 5242]. It was speculated that poly(ADP-ribosyl)ation of histone H1 and subsequent formation through crosslinking to an H1 dimer may be an important component of this phenomenon. To study this process, a method of complexing histone H1 to chromatin was required that promoted the restoration of accurate poly(ADP ribosyl)ation of this histone. Previously we have established that two histone H1 molecules are crosslinked by a chain of poly(ADP-Rib) 15 or 16 units in length. In the current study, we made use of the ability of oligonucleosomes, reconstituted with H1, to carry out the synthesis of the poly(ADP-Rib)-H1 complex in order to monitor the accuracy of reconstitution. It appears that a specific distance and juxtaposition of adjacent H1 molecules along the polynucleosome fiber is required for the enzymatic synthesis of this modified histone complex. We established that a controlled trypsin digestion of oligonucleosomes removed H1 histone with minimal perturbation of other nuclear proteins associated with chromatin. In addition, poly(ADP-Rib) polymerase was partially removed from chromatin by this procedure. Subsequently, methods utilizing gradient salt dialysis have been employed to reconstitute both the polymerase and histone H1 to the depleted oligonucleosomes. The reassociation of H1 (and polymerase) to specific binding sites within oligonucleosomes was accomplished by the above procedures. Poly(ADP-Rib)--H1-dimer synthesis was not observed in depleted oligonucleosomes, but this capacity was found to be partially restored in the reconstituted chromatin. Similarly, the ability of NAD to promote crosslinking of nucleosomes was restored in the reconstituted samples. These results provide a basis for further studies on how the poly(ADP-ribosyl)ation of histones alters the structure of chromatin. PMID- 6293829 TI - Effect of thiol reagents on the proton conductivity of the H+-ATPase of mitochondria. AB - The role of thiol groups in the proton conduction by the H+-ATPase of mitochondria is examined. A detailed kinetic analysis of the effect of arsenite and N-ethylmaleimide on the anaerobic relaxation of the proton gradient set up by respiration in 'inside-out' submitochondrial particles from beef-heart has been carried out. Arsenite, which reacts with vicinal dithiols, is shown to enhance the proton conductivity of the H+-ATPase. This effect is exerted on the F0 moiety of the complex and apparently mimics and is, in fact, favoured by a state of high proton conductivity induced in the complex by the respiratory delta mu H+. N Ethylmaleimide (MalNEt), which is a permeant monothiol blocking reagent, appears to attack critical -SH groups in a reaction leading to inhibition of the proton conductivity of the H+-ATPase. Also the inhibitory action of MalNEt on proton conduction is exerted on the F0 moiety of the H+-ATPase. Whilst the stimulatory effect of arsenite develops rapidly, the inhibitory action of MalNEt is sluggish and takes more than 10 min to fully develop. This and other kinetic characteristics, as well a partial additivity of the inhibition by MalNEt with that by oligomycin, indicate that the inhibitory action of MalNEt is associated to a substantial conformational transition in F0. Differences in the mechanism of inhibition of proton conduction by MalNEt and triphenyltin are also presented. PMID- 6293828 TI - Subclasses of simian-virus-40 large tumor antigen. Partial purification and DNA binding properties of two subclasses of tumor antigen from productively infected cells. AB - Two major subclasses of simian virus 40 tumor antigen were prepared from productively infected monkey cells. These subclasses can be distinguished by their sedimentation properties: one tumor antigen form sediments at 5-6S and the other at 14-16S. The DNA-binding properties of these subclasses were investigated by two different experimental procedures. In the first procedure, the DNA binding of subclasses of crude tumor antigen, separated by zone velocity sedimentation, were assayed by immunoprecipitation of the DNA-protein complexes. In the second procedure, the two tumor antigen forms were partially purified by column chromatography and DNA binding was tested in a filter binding assay. Both procedures gave comparable results. (a) The 5-6-S and the 14-16-S tumor antigen bound specifically to a DNA restriction fragment containing the viral genome control regions. (b) At low salt concentrations, both subclasses bound to specific and to nonspecific DNA sequences; competition experiments in the presence of nonspecific DNA showed, however, that the affinity of both tumor antigen forms for the viral genome control region was at least 10-fold higher than their affinity for nonspecific DNA sequences. (c) The binding of the 5-6-S subclass to viral control region DNA was optimal at 60-80 mM NaCl while specific DNA binding of the 14-16-S form was optimal at 150-200 mM NaCl; however, binding of the 14-16-S form to nonspecific DNA sequences was also more resistant to high salt concentrations than that of the 5-6S form. (d) Both tumor antigen forms bound well to specific and to nonspecific DNA at pH 6-6.5; with increasing pH values, binding to nonspecific DNA decreased while binding to specific DNA reached an optimum at pH 7-7.5. Binding of the 14-16-S form to viral origin DNA was more resistant to pH values above 7.5 than binding of the 5-6-S form. PMID- 6293830 TI - Cyclic AMP-dependent protein kinase in stimulated rat parotid gland cells: compartmental shifts after in vitro treatment with isoproterenol. PMID- 6293831 TI - Primary lymphoma of the thyroid gland: its limited diagnostic approach. PMID- 6293832 TI - Uremic neuropathy: correlations between electroneurographic parameters and serum levels of parathyroid hormone and aluminum. AB - An electrophysiological investigation on 29 dialysis patients was performed with the aim of verifying whether serum parathyroid hormone (PTH) and aluminum (Al) levels may play a role in the pathogenesis of uremic neuropathy. All patients except 1 showed electrophysiological abnormalities. Both PTH and Al levels were found to be significantly correlated with the time on dialysis. On the contrary, the values of each single electroneurographic parameter did not show any correlation with the duration of dialysis, PTH levels and Al levels. The authors conclude that PTH and Al does not seem to play a role in the etiology of uremic neuropathy. PMID- 6293833 TI - Adenylate cyclase activity and cAMP content of human platelets in uraemia. AB - We investigated the effects of uraemia and haemodialysis on the basal activity of adenylate cyclase and the cyclic-AMP content of human platelets in patients with end-stage renal insufficiency, patients receiving maintenance haemodialysis, and as controls healthy voluntary subjects. Basal adenylate cyclase activity in terminal renal disease (creatinine clearance less than 15 ml/min/1.73 m2) was 824 +/- SEM 57, in comparison to the healthy subjects with 453 +/-SEM 28 (P less than 0.001). We also found significant elevation (P less than 0 . 001) of platelet cAMP levels as compared to the controls. Basal adenylate cyclase activity and platelet cAMP levels were approximately normal in the dialysed patients. These results show that uraemic toxins adversely affect the platelet AC-cAMP system, possibly causing impaired platelet aggregation and the bleeding diathesis of uraemia. PMID- 6293834 TI - Pharmacokinetics of bacmecillinam and pivmecillinam in volunteers. AB - The pharmacokinetics of bacmecillinam and pivmecillinam were studied in healthy fasting volunteers given tablets in a cross-over, randomized order. The mean (+/- SD) peak levels of plasma mecillinam were 1.43 +/- 0.34, 2.73 +/- 0.43, and 4.62 +/- 1.41 mg/l after bacmecillinam 100, 200, and 400 mg and 2.38 +/- 0.65 mg/l after pivmecillinam 400 mg. The corresponding areas under plasma Vs time curves (AUC) were 2.21 +/- 0.19, 3.99 +/- 0.63, and 7.74 +/- 1.38 mg . h. l-1 for bacmecillinam and 5.35 +/- 0.93 mg . h. l-1 for pivmecillinam. The elimination half-lives were 0.8-1.1 h for bacmecillinam and 0.7 h for pivmecillinam. The 12 h urinary recovery of unchanged mecillinam after the 400 mg doses was 41% for bacmecillinam and 30% for pivmecillinam. The 400 mg dose of bacmecillinam gave a significantly higher plasma peak (p less than 0.001), AUC (p less than 0.001) and urinary recovery (p less than 0.001) than did pivmecillinam 400 mg. The plasma peaks appeared earlier and the rate of absorption was higher after bacmecillinam than after pivmecillinam (p less than 0.05). In conclusion, bacmecillinam had a better bioavailability than pivmecillinam in the tablet formulations studied. The AUC increased linearly with increasing doses of bacmecillinam. PMID- 6293835 TI - Distribution of oral ketoconazole to vaginal tissue. AB - Plasma samples and biopsies of vaginal tissue were obtained from 23 healthy women undergoing operative sterilization, 1 to 6 h after a single oral dose of ketoconazole 200 mg. Drug concentrations in plasma and tissue, were measured by a specific gas chromatographic method. The vaginal tissue concentration averaged 2.4 times less than the corresponding plasma levels. Equilibrium between tissue, and plasma was established within 1 h after dosing, when vaginal tissue levels exceeded 1 microgram/g. Ketoconazole concentrations decayed monoexponentially over the time interval studied (1-6h), with the similar half-lives of 1.2 and 1.4 H in plasma and tissue, respectively. Following an oral 200 mg dose, a tissue concentration not less than 0.01 microgram/ml was maintained over a 12 h period. This concentration has been shown to prevent outgrowth of the invasive (pseudo) mycelial form of Candida albicans. Hence, a b.i.d. or t.i.d. dosage schedule of ketoconazole in vaginal candidosis would give continuously effective levels at the site of infection. Ketoconazole concentrations in vaginal fluid are thought to be much higher than in the tissue because of ion-trapping. The present data may explain the efficacy of oral ketoconazole in the treatment of vaginal candidosis. PMID- 6293836 TI - Construction of a partial rabbit spleen cDNA library and identification of immunoglobulin clones. AB - A partial cDNA library was constructed from total poly A(+)-RNA isolated from the spleen of a rabbit (kappa allotype b5; heavy chain allotypes a3d12e15) that had been hyperimmunized with Streptococcus pneumoniae (type III). In spite of the absence of either specific DNA probes for rabbit immunoglobulin (Ig) sequences or cross-hybridizing mouse Ig DNA probes, recombinant clones containing cDNA sequences of rabbit gamma heavy chain and kappa light chains were identified by a combination of screening techniques: (a) colony hybridization using labeled mRNA; (b) mRNA hybridization selection and translation and (c) hybridization to electrophoretically fractionated poly A(+)-RNA ("Northern" analysis). Sequencing of three kappa light chain recombinant DNA sequences, including part of the 3' untranslated (UT) region, has confirmed the fact that recombinant DNA for kappa light chain mRNA has been identified. An unexpectedly high degree of homology between the 3' UT region sequence of this DNA from a rabbit of b5 allotype and the published 3' UT sequence from a b4 rabbit was found. It appears that 3' UT sequences from b4 and b5 alleles have diverged less than the coding sequences for the constant regions. The functional significance of this conservation of 3' UT sequences remains to be elucidated. PMID- 6293837 TI - Demonstration of slow channel activation in skeletal muscle of the dog. AB - The effects of verapamil, MnCl2, CaCl2 and tetrodotoxin on the contracture induced by KCl infusion were studied on the isolated, blood-perfused canine diaphragm preparation. Continuous intra-arterial infusion of KCl (30-100 mg/min) produced a biphasic contracture; initially a fast and large contracture was induced which soon declined and was replaced by a second smaller contracture which was sustained throughout the KCl infusion. Verapamil (3 micrograms-10 mg, i.a.) and MnCl2 (0.1-10 mg, i.a.) produced a dose-dependent relaxation in the sustained contracture. Intra-arterial injection of CaCl2 produced a dose dependent contractile response which was inhibited by treatment with verapamil and MnCl2. On the other hand tetrodotoxin, even in high doses, did not show a significant inhibitory effect on the sustained potassium-induced contracture. Therefore, the present results indicate that the slow calcium channel of skeletal muscle of the dog was activated with depolarization induced by potassium infusion. PMID- 6293838 TI - Changes in biogenic amine and benzodiazepine receptors correlated with conditioned emotional response and its reversal by diazepam. AB - Groups of littermate rats were trained to respond for food reinforcement on a variable interval one-min (VI 1) schedule, after which they were classically conditioned to associate a conditioned stimulus (CS) with footshock (conditioned emotional response; conditioned suppression; CER). Two control groups received yoked footshock (no CS) or the visual-auditory stimulus only (no footshock). On test day, a group of the CER conditioned animals received injections of either vehicle or diazepam prior to exposure to the VI 1 food-reinforced schedule. After 30 min of the VI 1 schedule, the CS was presented continuously for 15 min, after which the animals were decapitated, the brains removed, membranes prepared and in vitro receptor binding evaluated. During the CS, the CER animals suppressed responding and exhibited conditioned fear (emotional) behavior, while the control groups, and animals given acute diazepam, maintained normal responding. [3H]Diazepam binding was reduced in the CER animals, yet acute benzodiazepine administration did not effect this binding. [3H]QNB binding was reduced by CER and increased by diazepam administration. Adrenergic, serotonergic and dopaminergic systems were also evaluated. Traditional biogenic amine systems may respond to CER and diazepam administration in some compensatory manner. PMID- 6293839 TI - Pyrimido-pyrimidine derivative, RA642, a central pressor agent in cat endotoxin shock. AB - The effects of a pyrimido-pyrimidine derivative, RA642, in endotoxin shock were studied. Blood pressure and preganglionic splanchnic nerve (PSN) activity were measured in alpha-chloralosed cats. Within 30 min of intravenous administration of E. coli endotoxin (1 mg/kg), blood pressure and PSN activity were 68 +/- 8% and 47 +/- 10% of control and by 60 min were depressed by 54 +/- 8% and 42 +/- 8%. RA642 (0.25 mg/kg) injected i.v. 30 min after endotoxin, caused blood pressure to recover significantly to 83 +/- 6% of control within 5 min and be maintained at that level. PSN activity was similarly increased to and maintained at 62 +/- 9% of control. The efficacy of RA642 in reversing the lethal consequences of endotoxin shock were dramatic; all treated animals survived whereas the mortality rate of non-treated animals was 83%. This strongly suggests that the central pressor agent, RA642, may have important therapeutic applications in the management of endotoxin shock. PMID- 6293840 TI - Imidazole-induced potentiation of the contractile response to various agonists in vascular smooth muscle. AB - In isolated rabbit aortae, imidazole (10(-4)M) caused a unique nonspecific potentiation of the contractile responses to prostaglandins, norepinephrine, 5 hydroxytryptamine, histamine and potassium only at low concentrations. Imidazole had no effect on the dose-response relationship for Ca2+ in the presence of potassium, 40 mM. Imidazole potentiated contractions in supersensitive strips pretreated with reserpine or with low temperature (5 degrees C) for 5 days. In the presence of theophylline (3 x 10(-4)M), imidazole failed to potentiate the responses to norepinephrine, 5-hydroxy-tryptamine, histamine and potassium, but significantly potentiated responses to prostaglandin E1, E2 and F2 alpha. Imidazole (10(-4)M) significantly decreased the 45Ca uptake of aorta in a Ca-free solution, whereas the drug did not affect 45Ca uptake in a normal solution. These results suggest that imidazole-induced potentiation is related to an increase in Ca2+ permeability and possibly to an additive effect on Ca2+ binding. PMID- 6293842 TI - Captopril decreases vascular reactivity independently of changes in converting enzyme activity and prostaglandin release in the rat isolated kidney. AB - The relationship of the vascular effect of captopril to angiotensin converting enzyme activity and prostaglandin-dependent mechanism was studied in rat isolated kidneys, perfused with Krebs-Henseleit at 20 ml/min per 2 kidneys, with basal perfusion pressures of 78 +/- 1 mm Hg (Mean +/- S.E.M.). Two doses of captopril were used; both low (0.05 microgram/ml) and high doses (5 microgram/ml) inhibited maximally the vasoconstrictor responses to 100 and 200 ng of angiotensin I. Captopril at the low dose did not affect the renal vasoconstrictor responses to norepinephrine (NE) (25-400 ng), whereas high-dose reduced the vasoconstriction to all doses of NE. Treatment with captopril tended to diminish dose-related release of prostaglandins in response to NE. Indomethacin (1 microgram/ml) prevented NE-induced release of bioassayable and radioimmunoassayable prostaglandins but did not affect the ability of captopril to reduce NE-induced vasoconstriction. High-dose captopril also decreased the vascular reactivity to angiotensin II (5 ng) and lysine vasopressin (10 mU); however, the renal vasoconstriction caused by PGE2 (80 ng) was unaffected by captopril. We conclude that high-dose captopril decreased vascular reactivity by a mechanism independent of converting enzyme inhibition and unrelated to a prostaglandin-dependent vascular mechanism. PMID- 6293841 TI - Direct evidence that eseroline possesses morphine-like effects. AB - The opiate-like effects of eseroline, a physostigmine derivative, were studied in different tests. The antinociceptive effect of eseroline given s.c. and intracerebrally could be detected in the rat hot plate test and was reversed by naloxone. The apparent pA2 values of naloxone demonstrated with eseroline and morphine were found to be equal, suggesting an effect on similar receptors. Eseroline also had opiate agonist activity on the isolated longitudinal muscle strip of guinea pig ileum and isolated nictitating membrane of the cat: presynaptically, it inhibited the contractions evoked by stimulation and its effect was antagonized by naloxone. Eseroline reduced acetylcholine release from the myenteric plexus of the longitudinal muscle strip when the cholinesterases had been inhibited by physostigmine. In addition, it was also found that eseroline antagonized the inhibitory effect of normorphine in the longitudinal muscle strip and potentiated the effect of exogenous acetylcholine on smooth muscle, both effects being attributed to its anticholinesterase activity. The analgesic effect of eseroline was not related to its anticholinesterase activity. PMID- 6293843 TI - GABA abolishes cooperativity between benzodiazepine receptors. AB - Benzodiazepine receptors on neuronal membranes demonstrate cooperative interactions; these can be studied by observing the rates at which [3H]flunitrazepam dissociates from the receptors at different levels of receptor occupancy. In this study, the effect of GABA on the rates of reaction of [3H]flunitrazepam with benzodiazepine receptors is examined. GABA increases the association rate and decreases the dissociation rate of this ligand. In the presence of GABA, it is no longer possible to observe cooperativity between benzodiazepine receptors. PMID- 6293844 TI - Protection against chemically induced seizures by 2-amino-7-phosphonoheptanoic acid. AB - The anticonvulsant activity of 2-amino-7-phosphonoheptanoic acid (2APH) (an antagonist of excitation induced by N-methyl-D-aspartic acid) was studied against N-methyl-DL-aspartic acid (NMDLA), kainic acid, 3-mercaptopropionic acid (3MPA), thiosemicarbazide (TSC), quinolinic acid, bicuculline, picrotoxin and methyl-6,7 dimethoxy-4-ethyl-beta-carboline-3-carboxylate (DMCM) in Swiss S mice. 2APH, 0.33 mM/kg i.p., antagonizes convulsions induced by NMDLA, 3MPA, TSC, DMCM and picrotoxin but not those produced by the other convulsants. It is proposed that an aspartergic component may contribute to the development of convulsions after 3MPA, TSC, DMCM and picrotoxin. PMID- 6293845 TI - Modulation of the behavioral effects of amphetamine in rats by clonidine. AB - Clonidine (0.01, 0.05, 0.5 mg/kg) dramatically reduced the locomotor response to amphetamine (2 mg/kg) in a dose related fashion. In contrast, the same doses of clonidine had no effect on locomotions produced by a higher dose of amphetamine (6 mg/kg). Clonidine also had no effect on stereotyped head movements or the duration of the behavioral response to amphetamine. The lower dose of clonidine reduced amphetamine induced licking/biting while the two higher doses potentiated amphetamine induced licking/biting. Thus the behavioral effects of clonidine vary depending upon the dose of amphetamine and the particular behavior selected for study. The known neurochemical effects of clonidine do not account for this phenomenon. PMID- 6293846 TI - Inhibition of myocardial K+ channels by bromobenzoyl-methyladamantylamine, an adamantane derivative. AB - The effect of bromobenzoyl-methyladamantylamine (BMA) on transmembrane potentials and contractility of atrial and ventricular myocardium of guinea-pig and cat, as well as on transmembrane ionic currents of the frog atrial trabeculae was studied using conventional glass microelectrode and double sucrose-gap voltage clamp techniques. BMA markedly prolonged the action potential duration, depolarized the cell membrane, reduced the rate of rise of the action potential and exerted a positive inotropic effect on non-clamped myocardial preparations. The drug induced pacemaker activity in ventricular working muscle of cat. Moreover, BMA antagonized the effects of the K+ channel activator acetylcholine in a dose dependent manner. BMA was found to induce slow response action potentials in K+ depolarized ventricular myocardium of guinea-pig. In voltage clamp experiments, BMA reduced the outward K+ current but had no effect on either rapid inward Na+ or slow inward Ca2+ currents. The results suggest that BMA is capable of selectively blocking the myocardial K+ channels. PMID- 6293847 TI - Postsynaptic alpha 1-and alpha 2-adrenoceptor involvement in the vascular responses to neuronally released and exogenous noradrenaline in the hindlimb of the dog and cat. AB - The contribution of postsynaptic alpha 1-and alpha 2-adrenoceptors to vascular responses was investigated in the blood perfused hindlimbs of pentobarbitone anaesthetised dogs and chloralose-anaesthetised cats. Responses were obtained to sympathetic nerve stimulation, noradrenaline, phenylephrine and UK-14,304. In the dog and cat hindlimbs, the response to phenylephrine was reduced by the selective alpha 1-adrenoceptor antagonist, prazosin, while that to UK-14,304, in the dog, was reduced by the alpha 2-adrenoceptor antagonist, rauwolscine. Responses to noradrenaline and sympathetic nerve stimulation were only partially blocked by prazosin in both the dog and cat, but were further reduced by the addition of rauwolscine. These results suggest that the hindlimb vasculature of the dog and cat responds to neuronally released, as well as exogenous, noradrenaline by activation of both postsynaptic alpha 1- and alpha 2-adrenoceptors. PMID- 6293848 TI - A comparison of 3,4-diaminopyridine and 4-aminopyridine in the anaesthetized cat. AB - 3,4-Diaminopyridine and 4-aminopyridine were compared in the anaesthetized cat and found to be equiactive in their anti-curare activity. This in vivo similarity is at variance with previous in vitro studies which demonstrate 3,4 diaminopyridine to be more active than 4-aminopyridine at the neuromuscular junction. Possible reasons for the similarity between the two aminopyridines at the in vivo neuromuscular junction are discussed and it is concluded that 3,4 diaminopyridine has only marginal advantages over 4-aminopyridine as a potential anti-curare agent. PMID- 6293850 TI - Alpha 1-adrenoceptors induce Ca influx and intracellular Ca release in isolated rabbit aorta. PMID- 6293849 TI - Regional heterogeneity of benzodiazepine binding sites in rat brain. AB - Displacement of [3H]propyl-beta-carboline-3-carboxylate ([3H]PrCC) and [3H]flunitrazepam ([3H]FNM) was studied in rat hippocampus and cerebellum. Diazepam displaced both ligands with equal potency in both regions. Ethyl-beta carboline-3-carboxylate (beta CCE) and CL 218,872 displaced [3H]PrCC more potently than [3H]FNM and were more potent in the cerebellum than the hippocampus. Beta CCE and CL 218,872 interact more potently with BZ1 than BZ2 sites, while diazepam is equipotent. GABA and chloride ions enhance the potency of diazepam and CL 218,872 but not beta CCE. PMID- 6293851 TI - Purification of human melanocytes by monoclonal antibody combined with Percoll gradients. PMID- 6293852 TI - Phagocytotic activity of glial cells in culture. PMID- 6293853 TI - Isolation by tritium suicide of uridine-cytidine kinase-defective mutants in Chinese hamster V79 cells. PMID- 6293854 TI - Isolation of mouse primordial germ cells. PMID- 6293855 TI - Remarks on the differentiation of lysosomes from cultured human fibroblasts by silica gradient centrifugation. PMID- 6293856 TI - Effects of ATP, cAMP and pH on the initiation of flagellar movement in demembranated models of rat epididymal spermatozoa. PMID- 6293857 TI - Binding of plasma fibronectin to the surfaces of BHK cells in suspension at 4 degrees C. PMID- 6293858 TI - Virus proteins in herpetic keratitis. PMID- 6293859 TI - The role of herpes simplex virus secreted glycoproteins in herpetic keratitis. PMID- 6293860 TI - Raphe - cerebellum interactions. I. Effects of cerebellar stimulation and harmaline administration on single unit activity of midbrain raphe neurons in the rat. AB - The firing patterns of single raphe units at the posterior midbrain level were examined in chloralosed rats to assess the effects of cerebellar stimulation and/or harmaline administration. Raphe cells were grouped according to their spontaneous firing rate and other characteristics into two categories. From a total sample of 160 cells, 106 (66%) presenting a slow regular discharge pattern were classified as serotonergic (5-HT cells), whereas 35 (22%), having a faster firing rate, were considered non serotonergic (NS cells). Moreover, 19 (12%) raphe units were non categorized. Cerebellar juxtafastigial (JF) stimulation modified the discharge pattern of 56 (35%) raphe units. The remaining 65% were unaffected by the stimulation. Of the 41 5-HT cells affected by JF stimulation, 28 neurons (68%) showed a systematic increase of their firing rate, whereas of the 12 NS cells affected 8 neurons (66%) were inhibited. It thus appears that cerebellar stimulation has an opposite effect on raphe units according to the cell types. Harmaline administration suppressed the activity of 5-HT cells and increased the discharge rate of NS cells. Moreover, we noticed in the latter units a phase modulation of the firing pattern by pauses occurring with a fixed periodicity of 2.5 to 10 s. Considered in the context of previous studies, these results strongly suggest an inhibitory influence of the raphe system on the olivo cerebellar circuitry. PMID- 6293861 TI - Raphe - cerebellum interactions. II. Effects of midbrain raphe stimulation and harmaline administration on single unit activity of cerebellar cortical cells in the rat. AB - The effects of midbrain raphe stimulation and/or harmaline administration on cerebellar cell activities were examined in chloralosed rats. Cerebellar cortical cells were grouped into two categories. From a total sample of 68 cells, 48 were classified as Purkinje cells and the 20 others were unidentified. Midbrain raphe stimulation was found to inhibit for many sec the discharge of 40% of the Purkinje cells and 80% of the unidentified units. Other cells were unaffected, except 4 of them which were excited. Harmaline administration increased the CS firing rate of all Purkinje cells by inducing a rhythmic CS discharge at 7-12 Hz. Moreover, harmaline increased the discharge rate of unidentified units without inducing rhythmic activity. In both types of cerebellar cells a modulation of their firing pattern by periodical pauses at 0.1-0.4/s was noticed. These data are discussed in relation to the known influences of harmaline and cerebellar stimulation on raphe neurons. Taken in this context, results presented here confirm the existence of a modulatory influence of the raphe nuclei on the olivo cerebellar circuitry. A general model of interactions is proposed. PMID- 6293862 TI - The excitatory response of in vitro hippocampal pyramidal cells to normorphine and methionine-enkephalin may be mediated by different receptor populations. AB - As shown previously, opiate agonists increase the excitability of hippocampal pyramidal cells in a naloxone-reversible manner. In the present study, the degree of excitability was measured by population spike size recorded from hippocampus slices (CA1) obtained from naive or chronically morphinized rats. Cross tolerance could not be demonstrated to occur between met-enkephalin and normorphine in hippocampal cells made tolerant to morphine: the potent stimulatory effect of met enkephalin remains when applied to hippocampal slices removed from chronically morphinized rats, whereas normorphine was no longer effective. When these slices are washed or exposed to naloxone a diminution of the population spike occurs. These results suggest that while both opiate agonists increase neuronal excitability of hippocampal pyramidal cells, this effect is most likely mediated via different receptor populations. PMID- 6293863 TI - A high probability of an orientation shift between layers 4 and 5 in central parts of the cat striate cortex. AB - On the postlateral gyrus of the cat striate cortex the cells' preferred orientation was measured as a function of cortical depth in penetrations as parallel as possible to the radial fibre bundles. According to the penetration angle and in agreement with the current model of orientation columns, there was a low orientation drift in layers 2-4. At the transition between layers 4 and 5 an orientation shift of 45-90 deg was found in most penetrations. The orientation differences between adjacent recording sites in lower layers was normally low too, but clearly higher than in upper layers. The results are discussed in terms of more independent orientation mechanisms in upper and lower layers. PMID- 6293864 TI - ACTH and corticosterone secretion following insulin in intact and in variously hypothalamic deafferented male rats. AB - Adult male rats, intact (N) or with posterior (PHD), anterior (AHD) or complete (CHD) hypothalamic deafferentations, were injected with either 0.04 or 0.2 mu/100 g b.wt. of insulin. Forty-five minutes later they were decapitated and trunk blood was collected for serum glucose, adrenocorticotropic hormone (ACTH) and corticosterone (CS) determinations. The high insulin dose reduced serum glucose by approximately 50% and elicited a marked increase in serum ACTH and CS levels in all groups of animals as compared to the vehicle-treated group. In contrast, the low insulin dose which reduced serum glucose approximately 30% elicited a significant adrenocortical response only in the intact or PHD groups but failed to stimulate this response in animals with CHD or AHD. These results demonstrate that (1) CNS sites, outside the medial basal hypothalamus (MBH), mediate the adrenocortical response during the initial stages of hypoglycemia by a neural pathway impinging upon the CRF neurons from the rostral direction, and (2) the adrenocortical response during the more enhanced hypoglycemia stages is mediated by a systemic mechanism which acts directly on the MBH. PMID- 6293865 TI - Somatic sensory transmission to the cortex during movement: gating of single cell responses to touch. PMID- 6293866 TI - Somatic sensory transmission to the cortex during movement: phasic modulation over the locomotor step cycle. PMID- 6293867 TI - Altered cGMP-phosphodiesterase activity in chemotactic mutants of Dictyostelium discoideum. PMID- 6293868 TI - The design of a new group of angiotensin-converting enzyme inhibitors. AB - Using X-ray and NMR data relating to the conformation of the antihypertensive, angiotensin-converting enzyme inhibitor, captopril, and structure--activity relationships of analogues, it has been possible to postulate with the aid of computer graphics, the orientation of the three functions, the thiol, the terminal carboxyl and the carbonyl group which are involved in binding to the enzyme. Bicyclic mimetics of captopril, with related arrays of these functions, have been designed and synthesized. Compounds with the closest approximation to the array in captopril are the most active inhibitors of angiotensin converting enzyme, in vitro. PMID- 6293869 TI - Leukotriene-induced neutrophil aggregation in vitro. PMID- 6293870 TI - Involvement of calcium, calmodulin and phospholipase A in the alteration of membrane dynamics and superoxide production of human neutrophils stimulated by phorbol myristate acetate. PMID- 6293871 TI - Possible role of H+--alkali cation countertransport in secretory granule swelling during exocytosis. AB - Studies in model systems, as well as observations in intact cells, suggest that osmotic swelling of secretory granules is an essential step in exocytosis. A model is proposed whereby the low pH recorded in most secretory organelles could provide the driving force for granule swelling. The model assumes that during stimulation an exchange of H+ for alkali cations is triggered across the granule membrane. The outgoing H+ are rapidly replaced by the internal buffering capacity with a concomitant osmotic gain. The exchange is independent of anions and could be triggered by cytoplasmic Ca2+. The H+ -pump is responsible for the delta pH but independent of the exchange mechanism. PMID- 6293872 TI - Chemical modifications of pokeweed antiviral protein: effects upon ribosome inactivation, antiviral activity and cytotoxicity. AB - Pokeweed antiviral protein (PAP) is a protein known to inactivate eukaryotic ribosomes by an unknown enzymatic action and inhibit the production of mammalian viruses in tissue culture. This protein was subjected to a variety of chemical modifications to determine their effects upon ribosomal inactivation, antiviral action, and cytotoxicity. It was found that modifications of a number of different amino acid residues had similar effects upon all 3 activities. Also the inactivation of PAP with diethylpyrocarbonate was not due to its reaction with a histidine residue but to a modification of an unidentified amino acid residue. PMID- 6293873 TI - Parathormone promotes glycogen formation from [14C]glucose in cultured osteoblast like cells. AB - Parathyroid hormone stimulates [U-14C]glucose incorporation into glycogen of cultured osteoblast-like calvaria cells. This effect is detectable only several hours after the addition of PTH and it is mimicked by dibutyryl cyclic AMP. In contrast to insulin (in pharmacological concentrations), PTH enhances glycogen formation only in calvaria cells, but not in fibroblasts. Insulin-like growth factor I in physiological concentrations promotes glycogen-synthesis shortly after addition. PMID- 6293874 TI - A biotin-dependent sodium pump: glutaconyl-CoA decarboxylase from Acidaminococcus fermentans. AB - The decarboxylation of glutaconyl-CoA to crotonyl-CoA in the anaerobic bacterium Acidaminococcus fermentans is catalysed by a membrane-bound, biotin-dependent enzyme which requires Na+ for activity. Inverted vesicles from A. fermentans accumulated Na+ only if glutaconyl-CoA was decarboxylated. The Na+ uptake was inhibited by avidin but not by the avidin biotin complex. Detergents and ionophores such as monensin also prevented the Na+ transport. The results indicate that the enzyme is able to convert the free energy of decarboxylation (delta Go' approximately equal to -30 kJ/mol) into a Na+ gradient. PMID- 6293875 TI - The effect of sodium channel activators on muscarinic receptors of neuroblastoma cells. AB - Incubation of neuroblastoma NIE 115 cells with veratrine leads to an apparent reduction in the number of muscarinic acetylcholine receptors assayed by [3H]scopolamine methyl chloride binding. No true down-regulation of the receptors occurs but a component of veratrine with muscarinic receptor affinity, which is not veratridine, enters the intracellular water space during the incubation period and competes with [3H]scopolamine methyl chloride for the muscarinic binding sites in subsequent ligand binding assays unless it is carefully washed away. Treatment of cells with the agonist carbamoylcholine does, however, lead to a true downregulation of muscarinic receptors. PMID- 6293876 TI - Identification of a calmodulin-dependent glycogen synthase kinase in rabbit skeletal muscle, distinct from phosphorylase kinase. AB - A glycogen synthase kinase that is completely dependent on Ca2+ and calmodulin has been identified in mammalian skeletal muscle, and purified approximately 3000 fold by chromatography on phosphocellulose and calmodulin--Sepharose. The presence of 50 mM NaCl in the homogenisation buffer was critical for extraction of the enzyme. The calmodulin-dependent glycogen synthase kinase (app. Mr 850 000) is distinct from myosin light-chain kinase and phosphorylase kinase, but phosphorylates the same serine residue on glycogen synthase as phosphorylase kinase. The physiological role of the enzyme is discussed. PMID- 6293877 TI - Reconstitution of cytochrome bc1 complex into lipid vesicles and the restoration of uncoupler sensitivity. AB - Beef heart mitochondrial bc1 complex (ubiquinone--cytochrome c oxidoreductase) has been assayed by its ability to catalyse the reaction of duroquinol reduction of ferricytochrome c. When the isolated complex is reincorporated into lipid vesicles, the enzyme-catalysed electron transfer rate becomes uncoupler sensitive. Initial experiments suggest that a protonmotive force is generated across the vesicles when electron transfer is initiated. Both delta psi and delta pH components of this protonmotive force then influence an internal rate constant of the bc1 complex. PMID- 6293878 TI - Biosynthesis of cytochrome c oxidase by isolated liver mitochondria of the tadpole, Rana catesbeiana. PMID- 6293879 TI - Phosphorylation of myosin light chain in skeletal and smooth muscles. AB - Significant differences in the properties of myosin light chain phosphorylation in skeletal and smooth muscles may be important in considering the role of myosin phosphorylation in contraction. Repetitive, low-frequency stimulation of fast twitch skeletal muscle resulted in phosphorylation of myosin light chain. Some of the factors leading to phosphorylation under these conditions include 1) partial activation of myosin light chain kinase with each stimulus, 2) a slow rate (t1/2 = 1 s) of inactivation of the kinase activity, and 3) a very slow rate of dephosphorylation by myosin light chain phosphatase. Myosin light chain phosphorylation was correlated with potentiation of isometric twitch tension in posttetanic potentiation and staircase responses. Stimulation of contraction in bovine tracheal smooth muscle by the cholinergic agonist carbachol was correlated with phosphorylation of the myosin light chain. The initial rate and maximum extent of phosphorylation during the first minute of stimulation was dependent on the concentration of carbachol. Phosphate incorporation into light chain declined after 1 min whereas isometric tension was maintained. beta-Adrenergic inhibition of tension development was accompanied by a decrease in the rate and extent of phosphorylatable myosin light chain phosphorylation but was not associated with reduced affinity of myosin light chain kinase for calcium-calmodulin. PMID- 6293880 TI - Coordination of cardiac sarcoplasmic reticulum and myofibrillar function by protein phosphorylation. AB - Adrenergic stimulation alters functional dynamics of the heart by mechanisms most likely involving cyclic AMP (cAMP)-dependent protein phosphorylation. In vitro studies indicate that the myofibrils and sarcoplasmic reticulum (SR) may act as effectors of the adrenergic stimulation. cAMP-dependent phosphorylation of troponin I (TnI), one of the regulatory proteins of cardiac myofibrils, results in a decreased steady-state affinity of troponin C (TnC) for calcium, an increase in the off-rate for Ca2+ exchange with TnC, and a rightward shift of the relation between free Ca2+ and myofibrillar force or ATPase. Phosphorylation of phospholamban, a regulatory protein of cardiac SR, results in an increased velocity of Ca2+ transport by SR vesicles, an increased affinity of the transport protein for Ca2+, and an increased turnover of elementary steps of the ATPase reaction. These in vitro findings support the hypothesis that the inotropic response of the heart to catecholamine stimulation involves phosphorylation of TnI and phospholamban. Our in vivo studies with perfused rabbit hearts show that during the peak of the inotropic response to isoproterenol there is a simultaneous phosphorylation of TnI and an 11,000-dalton protein in the SR, most likely the monomeric form of phospholamban. PMID- 6293882 TI - Properties and function of phosphatases from vascular smooth muscle. AB - Myosin light chain phosphatase (MLCP) activity was present in extracts from a wide variety of mammalian tissues. A partially purified preparation of bovine aortic MLCP also showed activity against phosphorylase a and p-nitrophenyl phosphate (PNP). Whether these three activities are ascribable to a single multifunctional phosphatase or to three distinct phosphatases is unknown. The three phosphatase activities coelute during gel filtration both before and after treatment with ethanol showing exclusion volumes corresponding to 240,000 and 35,000 daltons, respectively. This indicates that the enzyme is dissociable into a smaller catalytic subunit. The widespread occurrence of MLCP activity and the close parallel among MLCP, phosphorylase a phosphatase, and PNP phosphatase activities suggest that the enzyme (or enzymes) may participate in physiological processes in addition to dephosphorylation of phosphorylated myosin light chains. PMID- 6293881 TI - Regulation of cardiac contractile proteins by phosphorylation. AB - Several of the contractile proteins of the heart can be phosphorylated, but in studies with isolated proteins only phosphorylation of the inhibitory subunit of troponin (TnI) produces a major change in the properties of the contractile system. As TnI is phosphorylated, the concentration of calcium required for activation of contraction is increased. Phosphorylation of the tropomyosin binding subunit of troponin (TnT) or of the light chain of myosin fails to change ATPase activity of the isolated protein system. Phosphorylation of TnI is stimulated by the beta-adrenergic system and inhibited by the cholinergic system. Maximum calcium-activated force produced by the contractile system can be increased in hyperpermeable cardiac cells by cyclic AmP (cAMP) or agents that stimulate cAMP synthesis. This change in the contractile system, which appears to be part of the physiological response to beta-adrenergic stimulation, is mediated by phosphorylation of an intermediate that then modifies the contractile system. Phosphorylation of the contractile proteins is not involved. PMID- 6293883 TI - Regulation of cardiac glycogen synthase. AB - In perfused rat hearts insulin can activate, and catecholamines can inactivate, glycogen synthase (EC 2.4.1.11); the magnitude of each hormonal response is magnified if tissue glycogen levels are depleted. Both beta-adrenergic and alpha adrenergic agonists inactivate insulin-stimulated and basal glycogen synthase, with each promoting the same extent of inactivation in both circumstances. In this system beta-adrenergic agonists act via cyclic AMP (cAMP), and alpha adrenergic agonists via Ca2+, whereas insulin action appears to be independent of either cAMP or Ca2+. The action on cardiac glycogen synthase by the physiological catecholamine epinephrine is apparently mediated by the concomitant interaction with both alpha and beta receptors; interaction with each is mediated by their separate second messenger systems, which combine to produce the end physiological response. PMID- 6293884 TI - Reversal of apparent premature ovarian failure in a patient with myasthenia gravis. PMID- 6293885 TI - Influence of human chorionic gonadotropin in vivo on steroid formation and gonadotropin responsiveness of isolated human preovulatory follicular cells. AB - Granulosa and thecal cells of preovulatory follicles taken from 12 women were isolated and incubated separately for 2 hours in the presence and absence of human chorionic gonadotropin (hCG). To six of these women, an ovulatory dose of hCG (9000 IU) had been given 24 to 30 hours before excision of the follicle. Following incubation, cellular cyclic adenosine 3':5' monophosphate (cAMP) levels and the medium content of progesterone (P), androstenedione (A), and 17 beta estradiol (E2) were determined. All follicles appeared healthy and well developed, and the oocytes recovered were morphologically normal and mature. Exposure to hCG in vivo caused a shift in steroidogenesis from A toward P formation in isolated thecal cells and a marked increase in the P production by the granulosa cells of the preovulatory follicles. Furthermore, the thecal cells, but not the granulosa cells, developed refractoriness to further stimulation with hCG in vitro. PMID- 6293886 TI - Gonadal function in men with testis cancer. AB - A group of 218 patients with unilateral germinal testicular cancer was investigated with biopsy from the contralateral testis and/or semen and hormone analyses after orchidectomy but before irradiation and chemotherapy. In 24% of the biopsies severe irreversible changes such as spermatogenic arrest, Sertoli cell-only tubules, hyalinized tubules, or carcinoma in situ were found. Serum testosterone (T) values were below the reference interval in 13% of the patients, whereas 12% had normal serum T values in combination with elevated serum luteinizing hormone (LH). Most serum follicle-stimulating hormone (FSH) values were above the reference interval. The impairment of gonadal function in the patients could be explained partly by the removal of one of the testes, by premorbid defects of spermatogenesis in the contralateral testis, and by elevated serum human chorionic gonadotropin (hCG) values caused by the cancer. PMID- 6293887 TI - Effect of prostaglandin D2 on gonadotropin release from rat anterior pituitary in vitro. AB - A new prostaglandin D2 (PGD2), which is highly concentrated in the rat hypothalamic-pituitary system, was investigated for its gonadotropin release and synthesis by radioimmunoassay (RIA). Significant increases in the release of luteinizing hormone (LH) and follicle-stimulating hormone (FSH) were observed when the anterior pituitaries were incubated for 4 hours in vitro in the presence of 500 micrograms PGD2/ml incubation medium. Little change in LH and FSH contents in the rat anterior pituitary was observed between the control and PGD2 stimulated groups during incubation. In addition, more than 200 micrograms PGD2/ml induced a marked increase of cyclic adenosine 3',5' monophosphate (cAMP) formation in the rat anterior pituitary, and this was observed within 10 minutes after stimulation by PGD2. These data indicate that PGD2 directly induces a marked release of both LH and FSH from the gonadotrophs, and the mechanism of action of PGD2 could be mediated by cAMP formation. PMID- 6293888 TI - [Response of pyramid neurons of the cerebral cortex to oxygen deficiency and cAMP administration]. AB - Contact microscopy of vital brain preparation revealed changes in bioelectric activity of the cat brain cortex during short--time anoxia following cAMP microionophoretic administration. cAMP at that acted as a protector of single pyramidal neurons as manifested in a decrement of "anoxia excitation" frequency and in an increasing number of previously "silent" neurons which could not be made active with an ordinary anoxia. The intracellular reductive equivalent accumulation rate also decreased essentially during anoxia. These effects depended on time interval after cAMP administration. PMID- 6293889 TI - [Effect of prostaglandin E2, dexamethasone, and exertion on adrenocortical and sympathetic adrenal activity in dogs]. AB - Influence of dexamethasone, prostaglandin E2 (PGE2) and physical load on activity of corticotropin, 11-oxycorticosteroids (11-OHCS), adrenaline, noradrenaline, glucose in arterial blood was studied in chronic experiment on dogs. A comparison with intact dog responses to exogenous PGE2 revealed an increase of corticotropin and 11-OHCS and a decrease of noradrenaline in arterial blood. Dexamethasone prevented adrenocortical activity in response to PGE2; it helped, however, to increase activity of noradrenaline. PMID- 6293890 TI - [Electrogenesis and contraction of smooth muscle taenia coli kept in a solution with elevated concentration of potassium ions]. AB - Experiments performed on smooth muscle strips of guinea-pig taenia coli in high potassium solution using sucrose-gap method revealed that phasic and tonic components of contractile response following potassium depolarization were due to influx into the cells of Ca++ ions from extracellular solution through two types of voltage-dependent Ca channels: the fast inactivated and the slow noninactivated those. After cessation of the fast Ca channel inactivation with anodal current the membrane recovers its ability to generate spontaneous or evoked AP depending on repolarization level. Under these conditions AP generation can also occur in Ca-free and Na-free solutions due to regenerative process of K channel activation. PMID- 6293891 TI - [Role of the adenylate cyclase system in utilization of succinate by the frog gastric mucosa]. AB - Interrelationship between the adenylate cyclase system state and utilization of succinate as a substrate of oxidation in mechanisms of HCl secretion by isolated gastric mucosa of frog Rana esculenta was studied. The inhibition of the protein synthesis by cycloheximide rendered gastric mucosa insensitive to the activation of HCl secretion by histamine and succinate. A similar effect was exerted by cimetidine, a blocking agent for H2-receptors. The blockade could be prevented with dibutyryl-cAMP. The seasonal dependence of activation of HCl secretion by specific and unspecific modifiers was discovered. The data obtained show the necessity of preliminary activation of adenylate cyclase and accumulation of limiting content of cAMP for the utilization of succinate in reactions of generation and transport of H+ by acid-producing cells of isolated gastric mucosa in frog. PMID- 6293892 TI - [Trigger function of cyclic AMP in the initiation of HCl secretion by the frog gastric mucosa]. AB - Histamine (0.3 mM) caused a rapid but transient increase in cAMP level prior to a rise in HCl secretion in the isolated gastric mucosa of frog Rana esculenta. H2 receptor blocking agent cimetidine (0.01 mM) suppressed HCl secretion which had been preceded by a decrease in cAMP level. No statistically significant changes in cGMP content were found. Dibutyryl cAMP imitated the effect of histamine. In frogs captured in spring, with a high level of HCl secretion and high content of cAMP, histamine exerted no stimulating effect. It seems to be the cAMP and not cGMP that acts either as a trigger in activation of HCl secretion or as a stabilizer of induced secretion. PMID- 6293893 TI - Intracellular levels of cyclic nucleotides and proliferative activity of V79-1A cells. AB - We studied intracellular levels of cAMP and cGMP in relation to the proliferative activity of Chinese hamster V79-1A cells. In the course of population growth the intracellular level of cAMP correlated with cell growth activity. The intracellular level of cGMP rather correlated with population density. Stimulation of stationary cells with fresh medium was accompanied by a surge of cAMP level while cGMP level was not affected. PMID- 6293894 TI - Effect of high concentrations of cAMP on proliferative activity of V79-1A cells. AB - We studied the mechanisms of the inhibitory effect of high cAMP concentrations on growth of Chinese hamster V79-1A cells. High cAMP concentrations inhibit growth of V79-1A cells in the S phase of the cell cycle. The inhibition is probably due to some effect on the metabolism of DNA precursors. We suggest that this inhibition is not induced by the cyclic nucleotide itself, but by AMP as a degradation product of cAMP. PMID- 6293895 TI - Inhibitory activity of cytosine arabinoside on Marek's disease virus. AB - The presence of ara-C in growth medium at concentrations of 10(-4) to 10(-7) M completely or partially suppressed the formation of plaques specific for MDV or HVT and decreased proportionally the growth of HPRS line 1 lymphoblastoid cells. Administration of ara-C to chickens immediately after infection with MDV (1 mg/chicken/day i.p. for 5 days) reduced the incidence of Marek's disease by 50%. Thus ara-C appears to be an inhibitor of Marek's disease. PMID- 6293896 TI - [Insulin action on adenylate cyclase. The effect of insulin on adenylate cyclase of rat fat cells in the presence of theophylline]. AB - Insulin-effects on adenylate cyclase activity, cyclic AMP (cAMP) content and free fatty acid (FFA) accumulation of rat epididymal fat cells were examined under conditions of maximal inhibition of phosphodiesterase by sufficient amounts of theophylline for the purpose of localizing the site of the antilipolytic action of insulin. After brief preincubation of the cells with physiological amounts of insulin in the presence or absence of 0.1 mM adrenalineee, fat cells were homogenized following the addition of theophylline and then further incubated. Under these conditions, small but significant enhancement of adenylate cyclase activity, cAMP content and FFA accumulation by insulin alone was observed, while insulin remarkably inhibited adrenalin-stimulated FFA accumulation, reducing adenylate cyclase activity and cAMP levels. This increase of cAMP content by insulin was only seen 5 or 15 minutes after the addition of theophylline. Furthermore, this insulin effect was also observed in the experiments which were performed in a medium containing high concentrations of albumin (2%). The concomitant accumulation of FFA might have resulted from the stimulation of lipolysis, rather than from the synthesis of FFA, since there was no added glucose in the medium. And finally, the hydrolysis of 14C-tripalmitate by a fraction of the cell homogenate under the presence of theophylline was more extensive after preincubation of the cells with insulin than without insulin. In summary, insulin, which is recognized as a typical antilipolytic hormone, activated adenylate cyclase and increased lipolysis at its physiological concentrations when it alone exerted its effect upon fat cells under the conditions where phosphodiesterase was completely inhibited by theophylline. Accordingly, the present results indicate the bimodal effect of insulin on adenylate cyclase and lipolysis under the presence of theophylline; enhancement when applied alone, and depression with adrenalin. So it is most likely that the "negative synergism" occurs as a net effect when a mild activator acts together competitively with a strong activator toward the same target. These data suggest the fundamental roles of adenylate cyclase systems in the mechanism of lipolysis regulation by insulin. PMID- 6293897 TI - Comparative studies on nucleoside diphosphatase of rat ascites hepatoma and rat liver: activity level, purification and properties. AB - 1. The activities of nucleoside diphosphatase in various rat ascites cells of hepatoma, and fetal and neonatal rat liver were much lower than that of normal adult rat liver. 2. The enzyme was purified from ascites hepatoma (AH-66 cell lines) to an apparently homogeneous state and the enzymatic properties were studied in comparison with the enzyme from rat liver microsomes. 3. The hepatoma enzyme had less stability based on the results of heat-inactivation experiments. 4. However, the other properties of hepatoma enzyme; Km value, molecular weight, optimal pH, isoelectric point, substrate specificity and antigenicity, were similar to those of rat liver enzyme. PMID- 6293898 TI - Human insulinoma hybrids produce proinsulin-like material. AB - We have established somatic cell hybrids by fusing cells from two human insulinomas with an established murine cell line LMTK- Cl1D. After selection of the hybrids, the media were analyzed and found to contain insulin and human C peptide immunoreactive material. The newly synthesized material was further characterized by pulse labeling and immunoprecipitation, and shown in five hybrid lines on Sephadex G-50 chromatography to have a size similar to proinsulin. The hybrids produced the apparent proinsulin-like material for up to 7 mo. Chromosome composition of the hybrids was determined by isozyme analysis and banding techniques. Chromosome 11, which previously has been assigned the insulin gene using cDNA probes, was identified in the hybrids producing proinsulin-like material. However, the retention of this chromosome did not always assure the production of hormone. This independent technique has confirmed the localization of the insulin gene to chromosome 11 and offers the opportunity of studying insulin processing and developing continuous insulin-producing cell lines. PMID- 6293899 TI - Responsiveness of plasma 18-hydroxycorticosterone and aldosterone to angiotensin II or corticotropin in nonazotemic diabetes mellitus. AB - To assess the function of the final step of the pathway for aldosterone biosynthesis, the responsiveness of plasma 18-hydroxycorticosterone and aldosterone concentrations to angiotensin II infusion was studied in 14 patients with nonazotemic diabetes mellitus as compared with 14 normal controls approximately matched for sex and age. In addition, the responses of both steroids to corticotropin injection were investigated in the diabetic patients. Under basal conditions, plasma aldosterone levels were slightly lower in the patients than in normal controls, while plasma 18-hydroxycorticosterone concentrations were similar in the two study groups. Angiotensin II induced marked and comparable increases in plasma 18-hydroxycorticosterone and aldosterone levels in normal and diabetic subjects. Plasma 18 hydroxycorticosterone and aldosterone levels before and after angiotensin II infusion were significantly interrelated; this correlation was similar in normal subjects (r = 0.61; P less than 0.001) and diabetic patients (r = 0.51; P less than 0.005). Plasma 18-hydroxycorticosterone and aldosterone were significantly increased by corticotropin in the patients. These findings indicate that the terminal step of aldosterone biosynthesis, which involves the production of 18 hydroxycorticosterone and aldosterone, is largely unaltered in patients with nonazotemic diabetes mellitus. PMID- 6293900 TI - Effect of gastrin on fasting and postprandial plasma GIP release in man. AB - In order to evaluate the mechanism behind the augmented postprandial gastric inhibitory polypeptide (GIP) release seen in patients with achlorhydria and hypergastrinemia, 8 healthy subjects were given a liquid test meal on three different days. On 1 day the meal was preceded by ranitidine (H2 receptor antagonist) to block the gastric H+ secretion. On another day the meal was also preceded by ranitidine, but on this occasion gastrin was infused intravenously during the first hour in order to induce hypergastrinemia. On a third day the meal was given along to serve as a control. Finally, 6 healthy fasting subjects were given a 1-hour intravenous infusion of gastrin after pretreatment with ranitidine. Plasma GIP responses after the meal on the days with ranitidine alone or together with the gastrin infusion did not differ significantly from that found on the control day. When gastrin was infused in the fasting state, no effect was seen on the basal GIP levels. Thus, neither exogenous gastrin nor achlorhydria seems to affect the plasma GIP release, and it appears more likely that a rapid gastric emptying rate can account for the augmented GIP response found in achlorhydric patients. PMID- 6293901 TI - Insulin secretion by fetal human pancreas in organ culture. AB - Whole fetal human pancreases of 12-22 weeks gestation, showed histological growth and differentiation in vitro over 3 weeks. At glucose concentrations of 1-4 g/l, there was no difference in insulin secretion into culture medium over 1 h. There was no stimulation of insulin release by D-glyceraldehyde, thus defective glucose stimulated insulin release was probably not due to impairment of an early step in glycolysis. In the presence of 0.5 mmol/l dibutyryl cyclic AMP, insulin secretion was enhanced (0.188 +/- 0.030 versus 0.100 +/- 0.012 mU x mg tissue-1 x h-1, p less than 0.001) independently of glucose concentrations. It thus appears that impairment of glucose-stimulated insulin release was unlikely to be due to insufficient intracellular cyclic AMP. Insulin release increased in response to tolbutamide and theophylline. Insulin secretion was stimulated in the presence of a fivefold increase in amino acid concentration (0.118 +/- 0.018 versus 0.031 +/- 0.008 mU x mg tissue-1 x h-1, p less than 0.001). There was a fourfold increase in basal insulin secretion from islets previously grown in high concentration of amino acids compared with standard culture medium, (0.284 +/- 0.052 versus 0.067 +/- 0.011 mU x mg tissue-1 x h-1, p less than 0.001), emphasizing the important role of amino acids as substrates for B cell metabolism and development. PMID- 6293902 TI - Origins of laboratory mice deduced from restriction patterns of mitochondrial DNA. AB - To determine the origins of laboratory mice, the restriction patterns of mitochondrial DNAs (mtDNAs) from various strains were compared with those of relevant subspecies and/or races of Mus musculus. In most strains and substrains of laboratory mice examined (50/55), the cleavage patterns were identical to those of the European subspecies M. m. domesticus. Those that varied include two sublines of NZB, the strain NZC, and the Japanese strain RR. The NZB and NZC patterns were identical to that of the European subspecies M. m. brevirostris, which itself has restriction patterns similar to M. m. domesticus. On the other hand, the RR pattern was identical to M. m. molossinus-like mice trapped in Western China and slightly different from Japanese M. m. molossinus. These findings suggest that the strains NZB and NZC stemmed from a European founder stock which differed from the ancestral stocks of other laboratory strains and that the ancestral mice of the RR strain had been transported from China to Japan. Therefore, most laboratory strains of mice are derived from the European subspecies M. m. domesticus while M. m. brevirostris and M. m. molossinus have made minor contributions. M. m. musculus does not appear to have made any contribution. PMID- 6293903 TI - Expression of embryonic haemoglobin in tsAEV-transformed embryonic erythroid cells during temperature-induced differentiation. AB - Cells prepared from 1-day-old chick blastoderms were infected with a temperature sensitive mutant of avian erythroblastosis virus (tsAEV). Clonal strains of transformed erythroblasts were isolated from the infected blastoderm cells. By shift to the nonpermissive temperature, these cells could be induced to differentiate into erythrocyte-like cells which expressed embryonic haemoglobins. Embryonic haemoglobins could not be detected in tsAEV-transformed erythroblasts from adult bone marrow when induced to differentiate under the same conditions. In contrast to normal primitive erythrocytes, tsAEV-infected embryonic erythroblasts differentiated in vitro expressed also adult haemoglobin. These results suggest an influence of the haematopoietic environment on the switch from embryonic to adult erythrocytes. PMID- 6293905 TI - [What is the role of hepatitis B virus in the appearance of hepatocellular carcinomas in patients with alcoholic cirrhosis?]. PMID- 6293904 TI - [New inotropic drugs]. PMID- 6293906 TI - The effects of vitamin E on the indicator enzymes of organella membranes in the injured liver. AB - Acute and chronic liver damage was caused by the administration of either galactosamine or carbon tetrachloride. Consequently, the rats with damaged livers were killed after vitamin E was administered. The livers were removed and were homogenated. Indicator enzymes (5'-nucleotidase, arylsulfatase, cytochrome C oxidase and glucose-6-phosphatase) of organella membranes were measured in the homogenates of the normal and damaged livers. The effects of vitamin E resulted in the stabilizing of the impaired membranes of plasma, lysosome, mitochondria and microsome; (1) the abnormal decrease of 5'-nucleotidase activity and glucose 6-phosphatase activity, and the abnormal increase of arylsulfatase activity, which induced galactosamine or carbon tetrachloride, and (2) the abnormal decrease of cytochrome C oxidase activity induced by galactosamine- HCl, were normalized. PMID- 6293907 TI - Cyclic adenosine monophosphate stimulates active potassium secretion in the rat colon. AB - To determine whether cyclic adenosine monophosphate influences active potassium transport in the rat colon, we studied the effect of dibutyryl cyclic adenosine monophosphate and theophylline on unidirectional transmural 42K fluxes across proximal colonic mucosa under short-circuited conditions. Active potassium secretion (-0.19 +/- 0.02 microEq/h X cm2) was present in animals maintained on a normal potassium diet. Both 0.5 mM dibutyryl cyclic adenosine monophosphate and 5 mM theophylline significantly increased net potassium secretion by 0.49 +/- 0.04 and 0.33 +/- 0.03 microEq/h X cm2, p less than 0.001, respectively; the stimulation of net potassium secretion was secondary to an increase in serosal-to mucosal potassium transport without change in mucosal-to-serosal potassium movement. A similar increase in active potassium secretion (from -0.15 +/- 0.03 to -0.32 +/- 0.03 microEq/h X cm2, p less than 0.005) was produced by bethanechol, a cholinergic muscarinic agonist that alters sodium and chloride transport by a noncyclic adenosine monophosphate, calcium-dependent process. In animals maintained on a high potassium diet, active potassium secretion was significantly increased to -0.79 +/- 0.17 microEq/h X cm2 (p less than 0.001). In these potassium-loaded animals, theophylline produced a greater increase in active potassium secretion (0.91 +/- 0.10 vs. 0.33 +/- 0.03 microEq/h X cm2, p less than 0.001) than in animals fed a normal potassium diet. These studies demonstrate that cyclic adenosine monophosphate and noncyclic adenosine monophosphate mediated secretogogues stimulate active potassium secretion. We speculate that the mechanism by which cyclic adenosine monophosphate increases active potassium secretion is related to an increase in luminal potassium conductance. PMID- 6293908 TI - Studies of the etiology of Crohn's disease using athymic nude mice. AB - Following injection of Crohn's disease tissue filtrates, lymphomas and hyperplastic lymph nodes developed in 16% of athymic nude (nu/nu) mice; whereas only 4% of control nude mice developed lymphadenopathy (p less than 0.025). One hundred forty coded sera from 111 patients (Crohn's disease = 36, ulcerative colitis = 28, diarrheal and other controls = 47) were assayed by indirect immunofluorescence for immunoreactivity with the lymphomas and hyperplastic lymph nodes. Coded sections were examined by two observers and scored on a 0 to 3 + scale. Fifty-four percent of the sera from patients with Crohn's disease were reactive with the Crohn's disease induced lymphoma by this assay. Eighty percent of sera from patients with symptomatic Crohn's disease were positive, whereas 22% of sera from patients in remission were positive. Sixty-six percent of sera from patients with symptomatic Crohn's disease reacted against hyperplastic lymph nodes induced by Crohn's disease filtrates. In contrast, only one control serum (from a patient with ulcerative colitis) reacted with the lymphomas or hyperplastic lymph nodes. Lymphomas induced by other means or arising spontaneously did not show immunofluorescence with Crohn's disease or control sera. Electron microscopy revealed C-type viral particles in five lymphomas induced by Crohn's disease filtrates and in one control lymphoma, but not in five hyperplastic lymph nodes and five control lymph nodes. Absorption of Crohn's disease sera with control lymphoma or with murine leukemia virus infected fibroblasts did not diminish immunoreactivity, whereas similar absorption with lymphomas induced by Crohn's disease filtrates abolished the immunofluorescence. These studies indicate that Crohn's disease tissue, when injected into athymic nude mice, induces lymphoid hyperplasia and lymphomas that contain an antigen(s) recognized by Crohn's disease sera. PMID- 6293910 TI - Cyclic AMP-dependent and independent positive inotropic effects of phenylephrine. AB - To summarize, a large number of studies indicate that the effects of phenylephrine on cardiac contractility are mediated through alpha-adrenoceptors, although not exclusively. There are an equally large number of reports indicating some beta-adrenoceptor stimulant effects of phenylephrine. Of the latter, some are based on the finding of antagonism by beta-receptor antagonists and some on the fact that phenylephrine is capable of elevating cyclic AMP levels, which is primarily a beta-, and not alpha-adrenoceptor effect. Although it is now widely accepted that phenylephrine elevates cyclic AMP levels, the contribution and significance of the elevated nucleotide levels to the overall positive inotropy of phenylephrine appears to be small. Alpha-Adrenoceptors may be involved with calcium, although the precise interaction between calcium and the alpha adrenoceptor is not clear. Finally, the adrenergic effects seem to depend on the experimental and physiological conditions including calcium ion concentration, thyroid state and driving frequency. PMID- 6293909 TI - [Rare presentations of cytosarcoma Phylloides]. AB - Three cases of cystosarcoma phylloides of the breast are presented. The different microscopic presentations are discussed. The therapeutic and prognostic importance of the histologic differences are discussed. Even after excision of the tumour in what appears to be benign types, recurrence must be expected. The recurrent tumour often shows a higher degree of atypia. The same conclusion is reached in other cases in the literature. An excision of the tumor can only be considered to be sufficient treatment if the microscopic findings show a benign well incapsulated tumour with few cells and the excision has been done well in healthy tissue. This should only take place in young women. Since cystosarcoma phylloides can usually not be differentiated clinically or mammographically from fibro-adenomas not even by cytology all fibroadenomas of the breast must be removed and subjected to microscopic evaluation. A mistake in the microscopic evaluation may arise if not several areas of the tumours are examined. The treatment of choice in cystosarcoma phylloides is removal of the breast especially if there is a marked epithelial component to the carcinosarcoma, axillary lymphadenectomy and possibly postoperative radiation may be necessary. PMID- 6293911 TI - Mode of inhibition of activity of Na+-K+-stimulated adenosine triphosphatase by indomethacin. AB - 1. The activity of the overall Na+-K+-ATPase reaction was inhibited by indomethacin in vitro. 2. The K+-NPPase activity was also inhibited by indomethacin. 3. The activity of Na+-dependent phosphorylation of Na+-K+-ATPase was activated by indomethacin. 4. Indomethacin required for 50% inhibition of K+ NPPase activity was 0.4 mM. 5. Inhibition mode of indomethacin for both the substrate and K+ in K+-NPPase reaction was competitive type. 6. The Ki values for indomethacin for the substrate and K+ in K+-NPPase reaction were 0.4 and 0.24 mM, respectively. 7. Inhibitory effect of indomethacin on K+-NPPase was reversible. PMID- 6293912 TI - Topoisomerase involvement in multiplicity reactivation of phage T4. AB - The products of phage T4 genes 39, 52 and probably 60 have been previously characterized as forming a type II DNA topoisomerase. Other evidence suggested that this topoisomerase promotes normal initiation of DNA replication, and that when it is defective its loss is partially compensated for by the host gyrase. We present evidence here that mutants defective in genes 39, 52 and 60 have reduced ability to carry out multiplicity reactivation (MR, a form of recombinational repair) of phage DNA damaged either by mitomycin C (MMC) or psoralen plus near-UV light (PUVA). We also observed that there is not extensive superhelicity in the intracellular phage DNA either in the presence or absence of the phage topoisomerase. This tends to rule out the possibility that the topoisomerase influences MR by controlling the general superhelicity of the phage DNA. The dependence of MR on topoisomerase could occur in several possible ways. However, we favor the explanation that the lesions are bypassed by a postreplication recombinational repair process that is influenced by the topoisomerase through its role in initiating replication. PMID- 6293913 TI - Site-specific instability in Drosophila melanogaster: evidence for transposition of destabilizing element. AB - In this study, we show that at least one lethal mutation at the 3F-4A region of the X chromosome can generate an array of chromosome rearrangements, all with one chromosome break in the 3F-4A region. The mutation at 3F-4A (secondary mutation) was detected in an X chromosome carrying a reverse mutation of an unstable lethal mutation, which was mapped in the 6F1-2 doublet (primary mutation). The primary lethal mutation at 6F1-2 had occurred in an unstable chromosome (Uc) described previously (LIM 1979). Prior to reversion, the fF1-2 doublet was normal and stable, as was the 3F-4A region in the X chromosome carrying the primary lethal mutation. The disappearance of the instability having a set of genetic properties at one region (6F1-2) accompanied by its appearance elsewhere in the chromosome (3F-4A) implies that a transposition of the destabilizing element took place. The mutant at 3F-4A and other secondary mutants exhibited all but one (reinversion of an inversion to the normal sequence) of the eight properties of the primary lethal mutations. These observations support the view that a transposable destabilizing element is responsible for the hypermutability observed in the unstable chromosome and its derivatives. PMID- 6293914 TI - Selfish DNA: a sexually-transmitted nuclear parasite. AB - A quantitative population genetics model for the evolution of transposable genetic elements is developed. This model shows that "selfish" DNA sequences do not have to be selectively neutral at the organismic level; indeed, such DNA can produce major deleterious effects in the host organism and still spread through the population. The model can be used to explain the evolution of introns within eukaryotic genes; this explanation does not invoke a long-term evolutionary advantage for introns, nor does it depend on the hypothesis that eukaryotic gene structure may be an evolutionary relic. Transposable genes that carried information specifying sexual reproduction in the host organism would favor their own spread. Consequently, it is tempting to speculate that some of the genes controlling sex were originally selected as transposable elements. PMID- 6293915 TI - Mutations in the pho80 gene confer permeability to 5'-mononucleotides in Saccharomyces cerevisiae. AB - Yeast mutants permeable to dTMP (tup) were selected and two new complementation groups (tup5 and tup7) were identified. Assay of the levels of both acid and alkaline phosphatase in cells grown under either repressing (5 mM PO4(-3) or derepressing (0.03 mM PO4(-3) conditions indicated that, in general, tup mutations cause cells to be defective in their regulation of phosphatase synthesis. In addition, three of the tup mutations (tup1, tup4 and tup7) displayed markedly elevated rates of inorganic phosphate transport. The tup7 locus was found to be tightly centromere-linked on the right arm of chromosome XV, and was shown to be allelic with the pho80 regulatory locus on the basis of both genetic and biochemical criteria. Analysis of other mutations known to affect phosphatase levels (pho) indicated that some also conferred permeability to dTMP. Possible allelic relationships between tup genes and certain of these pho mutations are discussed. Regardless of the culture conditions, wild-type strains were not permeable to dTMP; in contrast, it was found in the course of this work that normal yeast cells were permeable to dUMP and that dUMP permeability was regulated by the concentration of inorganic phosphate present in the medium used to grow the cells. Thus, permeability to 5'-mononucleotides appears to be under coordinate control with phosphatase synthesis. PMID- 6293917 TI - [Primary mapping of the Oag locus determining the synthesis of the somatic O antigen on the Vibrio eltor chromosome]. PMID- 6293916 TI - [Characteristics of hybrid plasmids carrying the genes of colicinogenic plasmid Collb-P9 responsible for colicin Ib synthesis and inhibition of phage T5 development]. AB - The EcoRI and HindII restriction endonucleases and pBR325 vector plasmid were used to obtain a set of hybrid plasmids containing ColIb-P9 fragments carrying the characters for colicin Ib synthesis and immunity and the ability to inhibit T5 phage growth. The genes responsible for colicin synthesis and immunity are closely linked and localized in the EcoRI fragment with a molecular weight of 1.85 MD (pIV41) or in the HindII fragment of 2.4 MD (pIV1). The clones containing these plasmids show an increased level of both spontaneous and mitomycin C induced colicin synthesis and an increased level of immunity due to a larger dosage of the genes. The genes controlling T5 growth inhibition are localized in other restriction fragments of ColIb DNA: the EcoRI fragment of 1.45 MD (pIV7) and the HindII fragment of 4.3 MD (pIV5). We have demonstrated by means of hybrid plasmids that T5 growth inhibition is not connected with the colicin Ib synthesized in infected cells and is controlled by other specific product(s) of the ColIb plasmid genes. T5 phage growth was as efficient in clones containing plasmids with cloned colicin Ib genes as in a strain without plasmids. An investigation of the expression of the genes inhibiting T5 phage growth in an in vitro protein synthesis system has revealed a protein with a molecular weight of 36 000 which seems to take part in the process. PMID- 6293918 TI - [Instability study of Escherichia coli strains containing hybrid plasmids with threonine operon fragments]. AB - The stability of Escherichia coli strains carrying hybrid plasmids which contain ColE1-like replicon and threonine operon genes was studied. It was shown that the main reason for instability is the loss of a plasmid. The second reason for instability is the rec-dependent recombination that leads to formation of new plasmids. All experiments where instability of strains was observed, can be quantitatively described by the model that presumes a random loss of plasmids in cell population with a frequency of about 7.10(-4), which results in diappearance of plasmid-borne cells due to their low growth rate. Instability increases during the stationary phase but it is not easy to quantitatively estimate this process. PMID- 6293919 TI - [Isolation and study of hybrid plasmids carrying Escherichia coli threonine operon genes]. AB - A fragment of Escherichia coli chromosome containing the intact threonine operon or its distinct genes has been cloned on the pBR322 plasmid. This fragment has been mapped using some restriction endonucleases. Cloning results in an increased level of appropriate enzyme activity in cells containing hybrid plasmids. Those carrying the complete threonine operon are capable of accumulating threonine up to 5 g/l in culture medium during 48 h. When multi-copy plasmids are used for gene cloning, interpretation of experiments aimed at transformation of auxotrophic bacterial strains, might be complicated. For example, transformation of appropriate threonine auxotrophs by a hybrid plasmid carrying mutation in the threonine gene, might result in prototrophic phenotype. It is possible that the great amount of mutant enzyme molecules compensated their low activity. On the contrary, the presence of a gene within the plasmid, as shown by restriction and biochemical analysis, did not always ensure the growth on a minimal medium of auxotrophs transformed by this plasmid. PMID- 6293920 TI - [Cloning of the adenylate cyclase gene of Escherichia coli K-12]. AB - The adenylate cyclase gene of Escherichia coli has been cloned on the plasmid vector pBR325. The hybrid plasmid pTH4 obtained has a molecular weight of 6,4 megadalton and represents pBR325 plasmid with the insertion of 2,8 megadalton in the Pst1 site. The cya mutant bacteria carrying pTH4 recover their ability to utilize mannitol, lactose and other carbohydrates as carbon sources, and lose this ability again in the case of rare spontaneous excision of the DNA insert from the Pst1 site. The phenotypical effect of pTH4 in cya mutants can be only seen in the crp+ genome. The strains carrying pTH4 are also characterized by the ability of beta-galactosidase induction under conditions of catabolite repression. Besides, the bacteria containing cya+ allele on the plasmid do not grow on glycerol, which seems to be caused by toxic concentrations of methylglyoxal formed as a result of the increased intracellular level of cyclic adenosine monophosphate. PMID- 6293921 TI - [Comparative analysis of the P-1-group plasmids of incompatibility]. AB - Wide host range plasmids (IncP-1) R906, R751 and R702 have several cleavage sites for BamHI, HindIII and EcoRI enzymes, in contrast to RP4 plasmid. Using these enzymes, deletion mutants of R906 plasmid have been obtained in vitro which only lost short DNA fragments (1 to 14 kb). A narrow host range pAV1 plasmid of the same incompatibility group has been transformed into the cells of Escherichia coli. pAV1 is stably maintained in the new host and retains its narrow host range in the course of conjugation. Different restriction fragments of R702, R751, R906 and R906-derived deletion mutants hybridize with the nick-translated probe of RP4 DNA. It is suggested that the wide host range plasmids have a similarity in structural and functional organization. PMID- 6293922 TI - [Amplification of genome regions in the somatic cells of mammals resistant to colchicine. III. Localization of the amplified genes in minute chromatin bodies]. AB - Small chromatin bodies (SCB) were revealed in Djungarian hamster cells resistant to colchicine. They looked like single bodies or like clusters of small particles. SCB were localized both in nucleus and cytoplasm. Similar formations were earlier observed in oocytes of insects with amplified extrachromosomal rDNA genes. DNA in the SCB was able to replicate not only during the S phase but also during other phases of the cell cycle. The restriction analysis showed that in cells with SCB DNA amplified sequences were replicated autonomously too. These data indicate that SCB in colchicine-resistant cells contain amplified genes. Besides, SCB double-minute chromosomes (DMs) were observed in some resistant sublines. In one of them, DMs were the only karyotypic alteration. The relationship between SCB, chromosomal homogeneously staining regions (HSRs) and DMs was studied. Single SCB and DMs appeared at the early stage of the development of colchicine-resistance (the level of drug resistance is 16-22). Selection of variants 170-220-fold resistant to colchicine was usually accompanied by the decrease in the cell number with SCB and DMs and by the increase in the amount of cells containing the chromosomes with HSRs. During the further enhancement of drug resistance (700-750), some decrease in the number of cells with HSRs and the appearance of the great number of cells containing large groups of SCB were found. The loss of colchicine-resistance observed during cultivation in colchicine free medium was accompanied by the disappearance of HSRs, emergence of SCB and DMs and further elimination of SCB and DMs from cells. The quantity of autonomously replicating amplified DNA fragments after digestive by HindIII was increased with the enhancement of SCB number in cultures. PMID- 6293923 TI - [Vibrio eltor mutants with altered hemolysin production: their isolation and characteristics]. AB - Hemolysin hyper- hypoproductive Vibrio eltor mutants and hemolytically inactive clones were isolated using tetracycline (Tn10) and chloramphenicol (Tn9) resistance transposons. Chromosomal localization was demonstrated for hml gene(s) involved in hemolysin production. Mutations responsible for an altered hemolysin production are likely to be situated closely to pur ura arg met chromosomal markers. PMID- 6293924 TI - Detailed physical mapping of the ribosomal RNA genes of Bacillus subtilis. AB - Characterization of patterns of ribosomal RNA (rRNA) homology with restriction digests of Bacillus subtilis 168 chromosomal DNA and with cloned DNA sequences has resulted in the construction of a physical map of the rRNA gene sets. There are two types of gene sets which differ in the size of "spacer" DNA sequences separating the 16S and 23S rRNA determinants. It was estimated that there are ten rRNA gene sets on the B. subtilis chromosome. PMID- 6293925 TI - The yeast frameshift suppressor gene SUF16-1 encodes an altered glycine tRNA containing the four-base anticodon 3'-CCCG-5'. AB - The SUF16 frameshift suppressor locus encodes a glycine tRNA. The SUF16-1 suppressor tRNA is inferred by DNA sequence analysis to contain the four-base anticodon sequence 3'-CCCG-5' in place of the wild-type anticodon 3'-CCG-5'. SUF16-1 mediates translation of the four-base messenger RNA (mRNA) sequence 5' GGGU-3' but apparently fails to act at the sequence 5'-GGGG-3'. A molecular model is presented that accounts for the observed specificity of tRNA-mediated frameshift suppression in Saccharomyces cerevisiae. PMID- 6293926 TI - An Mr 29000 protein is essential for mini-F maintenance in E. coli. AB - Plasmids consisting of mini-F inserted into multicopy vectors were constructed. Derivatives of these hybrid replicons were isolated which contained the transposon Tn5. The polypeptides encoded by these plasmids were identified by Escherichia coli minicell analysis. We show that a previously unidentified polypeptide of 29000 Mr is encoded by the mini-F gene E between 45.1 and 46.2 F kb on the mini-F plasmid map, and that this coding sequence (E gene) is transcribed rightward. Hybrid plasmids carrying Tn5 inserted into the E gene are unable to replicate in a polA- strain. Hence the E protein is essential for mini F replication. Mutations in the A and B genes of mini-F affect E gene expression, and the results suggest that E protein synthesis is stimulated by A protein. PMID- 6293927 TI - Partial restriction map of Marek's disease virus DNA. AB - A partial restriction map of Marek's disease virus (MDV) DNA was constructed by digestion with endonucleases BamHI, Bg/I and SmaI and by blotting hybridization. The data suggest that there is a terminal heterogeneous sequence at least on one end of the MDV DNA molecule. The data did not reveal four different orientations of the terminal fragments of MDV DNA molecules despite the observation that MDV DNA contains inverted repeat sequences as also present in Herpes simplex virus (HSV) DNA molecules (Cebrian et al., 1981). Terminal deletion of MDV DNA, SalI-H and I, was found in high passage number preparations. PMID- 6293928 TI - Cloning of the ColE3-CA38 colicin and immunity genes and identification of a plasmid region which enhances colicin production. AB - The colicin and immunity genes of plasmid ColE3-CA38 have been localized by characterization of bacteria carrying its cloned restriction fragments. They are within a 3.14-kb EcoRI segment, such that the immunity gene contains the KpnI site, and the colicin gene is adjacent to it within a 2.1-kb KpnI-HincII segment. The immunity gene and one end of the colicin gene are in the region of ColE3-CA38 which is not homologous to the closely related plasmid ColE2-P9. A 0.64-kb PvuI EcoRI segment of the plasmid adjacent to that containing the colicin and immunity genes was found to augment colicin production on solid media, and also affected the morphology of clearing zones produced by the cells when used as indicators in overlays of stabs of colicin E2 or E7 producers. The 0.64-kb segment was required in its native orientation relative to the 3.14-kb EcoRI segment to cause its effects. PMID- 6293929 TI - Representation of DNA sequences in recombinant DNA libraries prepared by restriction enzyme partial digestion. AB - We present a theoretical study of the fraction of sequences incorporated in a recombinant DNA partial digest library as a function of the size of the library. The fraction incorporated depends on the degree of restriction enzyme partial digestion. If all restriction sites in the target DNA can be cleaved with the same rate, optimum incorporation of sequences is observed when the number average length of the digested DNA equals the desired average length of the cloned insert. Overdigestion severely reduces the fraction of sequences present in a sample of clones. Heterogeneity in restriction enzyme cleavage rates also reduces the fraction incorporated, and underdigestion improves sequence representation in the face of cleavage rate heterogeneity. Practical methods for determining the number average length of partially digested DNAs are also presented. PMID- 6293930 TI - Escherichia coli plasmid vectors containing synthetic translational initiation sequences and ribosome binding sites fused with the lacZ gene. AB - The construction of a series of Escherichia coli plasmid vectors suitable for assaying the effects of gene control signals fused with the E. coli lacZ gene is reported. A synthetic deoxyoligonucleotide dodecamer 5'-CATGAATTCATG GTACTTAAGTAC 5' containing two translation initiation codons (ATG) separated by an EcoRI site was ligated with a lacZ gene derivative which lacks the codons for the first eight amino acids in plasmid pMC1403 (Casadaban et al., 1980). Two ribosome binding sequences were synthesised and inserted into the EcoRI site before an ATG, and the effects of these sequences on lacZ gene expression in vivo measured by assaying beta-galactosidase activity. The E. coli ribosomal RNA gene (rrnB) promoter, the tetracycline resistance gene promoter, and a lambda phage promoter were cloned using these plasmids. The plasmids are 9.9 kb in size, have ampicillin resistance as a selectable marker and are generally useful for the detection and in vivo assay of gene control regions. PMID- 6293931 TI - Cloning and characterization of a plasmid DNA from anacystis nidulans 6301. AB - A plasmid DNA of Anacystis nidulans 6301 was isolated by CsCl-EtBr centrifugation. The Mr of the plasmid, named pBA1, was estimated to be 5.04 +/- 0.26 X 10(6) by electron microscopic analysis and 5.2 X 10(6) by agarose gel electrophoresis. The pBA1 DNA was opened at a unique site with BamHI and cloned in pBR322 vector propagated in Escherichia coli HB101 cells. The recombinant plasmid, named pBAS18, was digested with various restriction endonucleases and its cleavage map was constructed. Based on this result, the cleavage map of the pBA1 plasmid is presented. PMID- 6293932 TI - A novel cloning vector for the direct selection of recombinant DNA in E. coli. AB - A new plasmid cloning vector (pHE3) is described carrying dominant p fluorophenylalanine-sensitivity (pheS) and chloramphenicol-resistance markers. This vector is used in combination with a p-fluorophenylalanine-resistant Escherichia coli recipient (strain RR28). Foreign DNA can be cloned into pHE3 leading to insertional inactivation of pheS. Transformation of RR28, and plating on minimal medium with chloramphenicol plus p-fluorophenylalanine (pfp) directly selects for colonies containing recombinant DNA. PMID- 6293933 TI - Restriction fragment maps of the genome of Bacillus subtilis bacteriophage SP beta. AB - Six different restriction endonucleases were used to generate restriction fragment maps of the genome of the temperate Bacillus subtilis phage SP beta. AvaI and SalI each had six target sites in the phage DNA, AvaII had three, BamHI had seven, PstI had twenty, and SacI had sixteen. Restriction analysis and heteroduplex analysis were used to locate a 10-kb region of DNA that is deleted in the clear-plaque mutant, SP beta cl. The deletion lay approx. 50 kb from the left end of the 126-kb phage genome. PMID- 6293934 TI - Technical assistance workshop for directors of NIMH-sponsored clinical training programs in mental health of the aging. PMID- 6293935 TI - [Glutathione content of the lens in various forms of cataract]. AB - Both the electrolyte ratio (K greater than Na) and high glutathione content of the lenses are generally undisturbed in the brunescent nuclear cataract (cataracta brunescens), the primary grey nuclear cataract, and the supranuclear cataract (senile cortical cataract). The electrolyte ratio (Na greater than K) and greatly decreased glutathione content are also grossly pathological in the subcapsular cataract (cataracta complicata, posterior subcapsular cataract), especially in connection with a secondary grey nuclear cataract, the mature cataract (cataracta matura), and the intumescent cataract. Alteration of the electrolyte concentrations and the glutathione content may be the results of physical, anatomical, and various metabolic disorders. PMID- 6293936 TI - [Data on the hygienic establishment of permissible residues for nitrogen nitrates in melon and vegetable corps]. PMID- 6293937 TI - [Decontamination of the area of acid burns of the skin contaminated with 239Pu]. PMID- 6293938 TI - [Substantiation of maximum permissible concentration of polidasol in the air of working zone]. PMID- 6293939 TI - Diet and gall stones: effects of refined and unrefined carbohydrate diets on bile cholesterol saturation and bile acid metabolism. AB - It has been suggested that consumption of refined carbohydrate foods (notably sugar and white flour) increases bile cholesterol saturation and hence the risk of cholesterol gall stone formation. To test this hypothesis, 13 subjects with probable cholesterol gall stones ate refined and unrefined carbohydrate diets, each for six weeks in random order. On the refined carbohydrate diet, subjects ate more refined sugar (mean = SEM: 106 +/- 7 vs 6 +/- 1 g/day, p less than 0.001), less dietary fibre (13 +/- 1 vs 27 +/- 3 g/day, p less than 0.001), and had a higher energy intake (9.17 +/- 0.66 vs 7.16 +/- 0.64 MJ/day, p less than 0.001). After each diet, the lipid composition of duodenal bile and bile acid kinetics was determined. The cholesterol saturation index of bile was higher on the refined carbohydrate diet in all but one subject, with a mean value of 1.50 +/- 0.10 compared with 1.20 +/- 0.12 on the unrefined diet (p less than 0.005). On the refined carbohydrate diet, bile contained relatively less cholic acid and slightly more deoxycholic acid. There were, however, no significant differences in total or individual bile acid pool sizes. There were also no differences in the rates of primary bile acid synthesis or fractional turnover on the two diets. Consumption of carbohydrate in refined form increases bile cholesterol saturation. The risk of gall stones might be reduced by avoidance of refined carbohydrate foods. PMID- 6293940 TI - Lack of association of cytomegalovirus antibody level with carcinoma of the uterine cervix. AB - Serum samples were collected from 199 patients attending a gynaecological clinic, colposcopy specimens being taken at the same time for routine pathology of paraffin sections which ranged from normal, through the degrees of dysplasia to carcinoma in situ. Other parameters noted were age, number of pregnancies and time since last pregnancy. When level of antibody to cytomegalovirus in each serum sample was tested by ELISA, anti-complement immunofluorescence and neutralisation, no correlation was found between antibody titre and pathological diagnosis. The only interdependence seen was that, as the age of the patients increased, so did the frequency of sero-positivity to cytomegalovirus. Also in the group studied the frequency of carcinoma in situ was not age dependent. PMID- 6293942 TI - Non-B, non-T acute lymphoblastic leukaemia terminating in malignant histiocytosis: a new case and a review of the literature. PMID- 6293941 TI - Restriction enzyme studies on highly repeated DNAs of human leukemic leucocytes. PMID- 6293943 TI - Cytochemical evidence of multiple molecular forms of myeloperoxidase (MPO). PMID- 6293944 TI - [Enhancement of platelet function in cholesterol-fed guinea-pigs]. AB - Effects of cholesterol-loading on platelet functions were studied. Guinea-pigs were fed on a diet containing 1% cholesterol for 1--5 months. Cholesterol-feeding caused an elevation of cholesterol content in the plasma and platelets, although the phospholipids increased only in the plasma. Enhanced platelet aggregation induced by ADP, collagen, and arachidonic acid was maintained for 5 months during cholesterol-feeding, concomitant with an increase in malondialdehyde production in platelets. Although basal levels of platelet cyclic AMP were not affected, PGE1-stimulated cyclic AMP levels were markedly decreased in platelets of cholesterol-fed animals. The same results were observed in platelets treated with theophylline. No change in the generation of PGI2-like substance by the aortic tissue occurred during 4 months of cholesterol-feeding, though a significant increase was seen after 5 months. These results suggest that the enhancement of platelet function by cholesterol-feeding is concerned with enhanced prostaglandin synthesis and suppressed adenylate cyclase activity in platelets. PMID- 6293946 TI - Collagenolytic activity of a soil actinomycete. PMID- 6293945 TI - Buffering capacity of the chief components of nutritive media for algae. AB - The buffering capacity of solutions of KH2PO4 and NaHCO3 increases with their concentration, the behaviour being describable by mathematical expressions. Solutions of KH2PO4 prepared from tap water exhibit a buffering capacity higher by an order of magnitude than those prepared from distilled water. However, there is no difference between the buffering capacities of solutions of NaHCO3 prepared from tap and distilled water. Urea and NH4NO3 have almost no effect on the buffering capacities of solutions. PMID- 6293947 TI - [Computer tomography of the thorax in the Poland syndrome]. PMID- 6293948 TI - [Hepatitis B vaccines. An evaluation of human plasma extracted vaccines]. PMID- 6293949 TI - [Ranitidine--what will the future bring]. PMID- 6293950 TI - An autopsy case of primary hepatoma associated with an oral contraceptive. AB - Primary hepatoma was found in a 38-year-old female who had been using an oral contraceptive for 28 months. Histologically the hepatoma was a well differentiated hepatocellular carcinoma. Alpha-fetoprotein was not increased and tests for HBS antigen was negative. Carcinoembryonic antigen was elevated remarkably before death. The findings of hepatic arteriography and peritoneoscopy suggested metastatic tumors of the liver rather than primary hepatoma. During the course of the disease, phlebothrombosis occurred and spread widely in the lower left limb. The observation of erythrocytosis indicated the presence of a tumor producing erythropoietin or an erythropoietin-like substance. PMID- 6293951 TI - [Underlying mechanisms of inotropic effects mediated by alpha-adrenoceptors in mammalian left atria--influences of stimulation frequencies and Ca++ channel blocking agents--]. AB - Experiments were performed in the electrically driven left atria of guinea pigs, rabbits and rats in order to clarify the inotropic mechanisms of myocardial alpha adrenoceptor stimulation. In the left atria of guinea pigs stimulated of 0.2 Hz, stimulation of alpha-adrenoceptors produced triphasic time response curves which composed of an initial positive inotropic phase followed by a negative phase and then a second positive phase. The negative inotropic effect leading to the occurrence of the triphasic time course was depend on concentration of phenylephrine and stimulation frequency, i.e. it became more manifest with increase in the concentration of phenylephrine and with decrease in stimulation frequency. Phenoxybenzamine blocked both the positive and the negative phase. Phentolamine and yohimbine preferentially inhibited the negative phase while prazosin preferentially depressed the positive phase. These results suggest that two populations of alpha-adrenoceptors might be present; one stimulatory and one inhibitory. In the left atria of rabbits, alpha-adrenoceptor stimulation increased the contractile force at rates more than 0.5 Hz, whereas the same procedure resulted in a negative inotropic effect at 1 beat/min. Rested state contractions were significantly decreased by alpha-adrenoceptor stimulation, while increased by beta-adrenoceptor stimulation. Post rest contractions, when NaCl concentration was reduced to 50%, was significantly diminished by alpha adrenoceptor stimulation in contract with significant increase by beta adrenoceptor stimulation. In the left atria of rats, alpha-adrenoceptor stimulation did not strongly influence a negative staircase whereas beta adrenoceptor stimulation abolished it. In the partially depolarized preparations, alpha-adrenoceptor stimulation did not always restore the mechanical responses. Lower concentration of Ca++ channel blocking agents, which are known not to inhibit the slow inward current, significantly depressed the alpha-mimetic positive inotropic effect. These results indicate that the positive inotropic mechanisms of alpha-adrenoceptor stimulation might be closely related to the surface Ca++ rather than to an enhancement of the slow inward current. PMID- 6293952 TI - Effect of dexamethasone on steroidogenesis and on adrenocortical cyclic AMP and cyclic GMP responses to ACTH. AB - The inhibitory action of dexamethasone on the adrenal steroidogenic response to ACTH was confirmed by im administration of graded doses (5, 10 and 30 ng) of synthetic beta 1-24 ACTH to young adult male rats which had received dexamethasone (0.1 mg/100 g bw) 4 hr prior to sacrifice. Following this, kinetic studies were performed by measuring plasma corticosterone, adrenocortical cyclic AMP and cyclic GMP before and 4, 12 and 30 min after administration of either 10 or 30 ng of ACTH. These doses were selected because their effects could be either completely or partially inhibited by dexamethasone. In rats without dexamethasone all the doses of ACTH which were checked induced an increase in both corticosterone and cyclic AMP and a decrease in cyclic GMP. With the smallest dose of ACTH the earlier administration of dexamethasone resulted in complete suppression of both the steroidogenic response and the cyclic AMP response. With the largest dose of ACTH both responses were diminished. In dexamethasone-treated rats the decrease in cyclic GMP was significantly less pronounced 4 min after ACTH than it was in non-treated rats. These results support the view that cyclic AMP and cyclic GMP might both be concerned with the mechanism of acute adrenal steroidogenesis. PMID- 6293953 TI - Retroviruses and embryogenesis, Seventh Adolf Butenandt Lecture given to the Gesellschaft fur Biologische Chemie in Damp 2000 on September 29, 1982. PMID- 6293954 TI - Effects of environmental stress or ACTH treatment during pregnancy on maternal and fetal plasma androstenedione in the rat. PMID- 6293955 TI - Growing pains. PMID- 6293956 TI - A 74-year-old with 'recurrent varices'. PMID- 6293957 TI - Cutaneous cytomegalovirus vasculitis: an unusual clinical presentation of a common opportunistic pathogen. AB - A case of cutaneous leukocytoclastic cytomegalovirus (CMV) vasculitis arising in a man with acute myelogenous leukemia is described. An antemortem biopsy specimen of ulcerated skin and an open lung biopsy specimen showed leukocytoclastic vasculitis and nonspecific diffuse interstitial pneumonitis, respectively, neither tissue demonstrating viral infection. Autopsy material revealed CMV vasculitis with typical intranuclear inclusions identified in enlarged endothelial cells associated with thrombus formation and luminal narrowing, in addition to florid CMV pneumonitis. This case represents an unusual although clinically relevant expression of a common opportunistic pathogen. PMID- 6293958 TI - The DNA tumor virus SV 40 induces gene mutations in human cells. Reversion of HPRT deficiency. AB - The mutagenic effect of papovavirus SV40 on human cells could be demonstrated by reversion of HPRT deficiency in Lesch-Nyhan fibroblasts transformed by the virus. SV40 seems to induce different gene mutations in individually selected cell clones, as was clearly shown by the respective HPRT enzyme properties. The consequences of our results for use of SV40-derived vectors in gene substitution experiments are discussed. PMID- 6293959 TI - Comparison of the inhibitory action of delta-9-tetrahydrocannabinol and petroleum spirit extract of herbal cannabis on human sperm motility. AB - 1 Using the trans-membrane migration method to measure drug effects on human sperm motility, it is found that delta-9-tetrahydrocannabinol is the major constituent of cannabis which inhibits sperm motility and unlike the cataleptic effect, other constituents in the petroleum spirit extract do not potentiate this effect significantly. 2 It is suggested that the mechanisms for sperm immobilization and cataleptic effects are different. PMID- 6293960 TI - The pulmonary macrophages of human smokers: decreased surface Fc receptor concentrations on heavily particle-laden cells. AB - 1 Human pulmonary macrophages (PM) have been obtained by broncho-pulmonary lavage from smokers and non-smokers. Smokers' PM tend to be larger cells with more particulate intracytoplasmic inclusions than non-smokers' cells. 2 Electronmicrographs show that the majority of inclusions are electron dense and within lysosomes. Other lysosomal inclusions in smokers' PM are needle-like structures and lamellar bodies. 3 A rosette assay that detects changes in surface Fc receptor expression was used to compare the Fc receptor expression of a PM subgroup that was heavily particle laden with a 'clean' PM subgroup. 4 There is strong evidence for reduced surface Fc receptor expression in heavily particle laden PM. The possible explanations of this difference are discussed. PMID- 6293961 TI - Establishment and characterization of chicken embryo fibroblast clone LSCC-H32. AB - Cell line CEC-32 and clone LSCC-H32 were established from primary chicken embryo cells spontaneously but not experimentally transformed at 32 degrees C. The lines consisted of fibroblastoid and polygonal cells and had a subtetraploid karyotype of 2N = 130 to 140. The cells showed increased plating efficiency and metabolic activities as demonstrated by hexose uptake and plasminogen activator assay. The established cells produced avian lymphoid leukosis viruses of subgroups A and B. The virus released from LSCC-H32 cells induced lymphoid leukosis in inoculated chickens 18 to 22 wk post infection (PI). The cells have been carried in continuous culture for 285 passages and they appeared to grow indefinitely. They were efficiently used to propagate several animal viruses and to titrate chicken interferon. PMID- 6293962 TI - In vitro isolation and establishment of new embryonal cell lines from rat nephroblastoma. PMID- 6293963 TI - Cell culture and in vitro studies of fresh and cryopreserved human insulinoma. AB - Dispersed cells from both fresh and cryopreserved human insulinoma have been maintained in cell culture. Initial yield of viable cells was 50% for fresh and 25% for cryopreserved tissue. Viability of cells in culture was documented by increasing numbers of cells (doubling time approximately 5 d initially and 2 d at the sixth subculture for both fresh and cryopreserved tissue) and continued release of insulin over time (approximately 100 ng/ml per 10(5) cells at 10 d and 175 ng/ml per 10(5) cells at 30 d of culture for both fresh and cryopreserved tissue). Evidence that cells growing in culture were beta cells was provided by: (a) recovery of intracellular and extracellular immunoreactive insulin (IRI), (b) electron microscopic morphology, and (c) immunohistochemical staining. Cells from fresh insulinoma incubated with increasing concentrations of extracellular glucose released increasing amounts of IRI up to approximately 15 mM glucose, which paralleled changes in plasma insulin obtained during a preoperative glucose tolerance test. PMID- 6293964 TI - The adverse effects of HEPES, TES, and BES zwitterion buffers on the ultrastructure of cultured chick embryo epiphyseal chondrocytes. AB - Chick embryo epiphyseal chondrocytes cultured in media containing HEPES, TES, and BES zwitterion buffers, used in combination or independently, consistently developed cytoplasmic vacuoles. This cytoplasmic vacuolation was resolved when the zwitterion buffered media was replaced by media containing bicarbonate:CO2 enriched air buffer. Vacuoles were infrequent or absent in cultures grown in bicarbonate:CO2 enriched air. Chondrocytes with an established extracellular matrix showed less vacuolation than fibroblastlike and polygonal shaped cells that lacked such a matrix. The granular endoplasmic reticulum and Golgi dictyosomes of zwitterion buffered chondrocytes were distended and contained a flocculent amorphous material. Cytoplasmic vacuoles (0.5 to 3.0 micron diam) formed by the fusion and intracellular accumulation of Golgi vesicles and vacuoles also contained a flocculent material enhanced by ruthenium red. Membrane bound extracellular vacuoles containing ruthenium red stained proteoglycan aggregates were common in the extracellular matrix of zwitterion buffered cultures but were generally absent from bicarbonate treated cultures. Electron dense calcium deposits seemed much larger and more numerous in the presence of zwitterion buffers. It is suggested that HEPES, TES, and BES buffers, used alone or in combination, may adversely affect cell membrane systems, and thus the transport or secretory mechanisms operative in cultured chondrocytes, or both, resulting in vacuole formation and the intracellular accumulation of synthesized export material. Although the mechanism by which HEPES, TES, and BES induce these changes remains unclear, the use of zwitterion buffers in biological preparations should be treated with caution. PMID- 6293965 TI - Transport of ions across an artificial membrane incorporated with some phospholipids. PMID- 6293966 TI - Histomorphologic spectrum of tumours of minor salivary glands. PMID- 6293967 TI - Clonal analysis of a human lymphoblastoid cell line (B17) secreting antibody to N acetyl-D-glucosamine. AB - In this paper we analyse the clonal composition of a human lymphoblastoid B-cell line secreting IgM/k antibody to N-acetyl-D-glucosamine, the immunodominant sugar of Group-A-streptococcal carbohydrate. Besides non-antibody secreting cells, the line consists of two clonotypes of antibody-secreting cells: B17 cells producing over 90% and F6 cells producing less than 10% of the antibody in the supernatant. The proportions of B17 and F6 cells in the cell line seem to be similar to the proportion of antibodies in the supernatant. F6 cells can be isolated by cloning and maintained as stable lines, whereas this is more difficult with B17 cells. The results suggest that upon establishment of the line, at least two N-acetyl-D glucosamine-specific B cells were immortalized and coexist together as independent clonotypes. Although F6 cells seem to have a slight tissue culture advantage, they represent the minor clonotype in the B17 cell line. PMID- 6293968 TI - Early changes in cyclic AMP and calcium efflux during phagocytosis by neutrophils from normals and patients with chronic granulomatous disease. AB - During phagocytosis of serum-treated zymosan particles (STZ) human neutrophils respond within 5 sec by a two-fold increase in cAMP concomitantly with an increase in 45Ca-efflux from intracellular stores. The changes in cyclic AMP and calcium efflux during phagocytosis were essentially the same whether the cells were preincubated in the presence of 5 mM glucose or without glucose. However, the phagocytic capacity, and oxygen consumption and degranulation during phagocytosis were reduced about 20%-25% in the absence of glucose. Addition of 2 deoxyglucose and iodoacetamide, which results in depletion of cellular ATP, abolished the cAMP increments during phagocytosis and profoundly inhibited calcium efflux. At the same time, the phagocytic capacity and STZ-induced oxygen consumption and degranulation were severely impaired. The mitochondrial inhibitors, cyanide, azide, and antimycin A, in the presence of glucose, did not affect the cellular ATP levels. Neither were cAMP increments and calcium effluxes during phagocytosis affected by these drugs, and likewise these did not affect phagocytosis or the post-phagocytic events. Neutrophils from two patients with X linked chronic granulomatous disease (CGD) and one patient with the autosomal recessive form elicited normal increases in cAMP levels and calcium effluxes after addition of STZ despite no increase in oxygen consumption showing that these early metabolic events are not affected by the lack of oxidase function. PMID- 6293970 TI - In vitro induction of anti-intermediate filament antibody in lymphocyte cultures by Epstein-Barr virus. AB - Serum antibodies reactive with intermediate filaments of the cytoskeleton (anti IF antibodies) are often present in infectious mononucleosis, some other viral diseases, and rheumatoid arthritis. The mechanism of their production is not known, but it is possible that the formation of this and other autoantibodies result from polyclonal activation of B-cells. Peripheral blood mononuclear cells from subjects with or without serum anti-IF antibody were therefore cultured in the presence or absence of Epstein-Barr virus (EBV). IgM anti-IF antibody was produced in both unfractionated and T-cell-depleted cultures, but not in the supernatants of the same cells cultured without added EBV. PMID- 6293969 TI - Conversion of leukotriene D4 to leukotriene E4 by a dipeptidase released from the specific granule of human polymorphonuclear leucocytes. AB - Leukotriene D4 (LTD4), the most active spasmogenic leukotriene constituent of the slow reacting substance of anaphylaxis was converted by suspended human polymorphonuclear leucocytes (PMNs) to a single, less polar metabolite which was not further catabolized. This product was identified as leukotriene E4 (LTE4) by its retention time during reverse phase-high performance liquid chromatography (RP-HPLC) and subsequent bioassay on the guinea-pig ileum. LTD4 with a retention time of 21 +/- 1.6 min (mean +/- SD) and a contractile activity of 5.0 +/- 0.4 u./pmol (mean +/- SD) was quantitatively converted extracellularly by PMNs to LTE4 with a retention time of 26 +/- 1.8 min and a contractile activity of 1.2 +/ 0.3 u./pmol. Subcellular fractionations of PMNs revealed the recovered LTD4-to LTE4 converting activity, termed LTD4 dipeptidase, to be localized only in he granule fraction. There was a time- and calcium-dependent extracellular release of LTD4 dipeptidase in association with lysozyme (r = 0.97, n = 16, P less than 0.001), a constituent of both specific and azurophilic granules, in the absence of release of cytoplasmic lactate dehydrogenase (LDH) and of beta-glucuronidase from the azurophilic granule. Phorbol myristate acetate (PMA), which selectively induces secretion of specific granules, released lysozyme and the LTD4 dipeptidase in a constant dose-dependent manner from PMNs (r = 0.96, n = 8, P less than 0.001). Calcium ionophore A23187 at concentrations less than 10(-7) M stimulated the parallel secretion of LTD4 dipeptidase and lysozyme (r = 0.91, n = 9, P less than 0.005), dipeptidase and lysozyme (r = 0.91, n = 9, P less than 0.005), whereas higher concentrations resulted in secretion of beta-glucuronidase and additional lysozyme without further release of dipeptidase. Thus, human PMNs can convert LTD4 to LTE4, a less vasoactive and spasmogenic leukotriene, via the secretion of a dipeptidase associated with the specific granules. PMID- 6293971 TI - Effect of quartz dust on peritoneal exudate cells in rat. PMID- 6293972 TI - Aetiology of the 1978 outbreak of encephalitis in Tirunelveli and other districts of Tamil Nadu. PMID- 6293973 TI - [A rapid method for the determination of virus-specific IgM antibodies]. AB - A newly developed ion exchange chromatography is reported; it is a simple and quick method to separate a highly purified IgM fraction from blood serum. The immunofluorescence test was used to demonstrate that these fractions contain only virus-specific IgM antibodies, but no demonstrable IgG antibodies, which can often block the specific IgM reactivity by competitive inhibition. The specificity of the reaction was tested on sera from 145 persons: the IgM test was always positive in clinically proven viral infections; the reaction was negative in all patients whose infection had occurred some time in the past. A series of suspected virus infections could be verified by the demonstration of specific IgM antibodies. PMID- 6293974 TI - Epidemiological aspects of acute viral hepatitis in drug abusers. AB - The epidemiological features of acute symptomatic viral hepatitis were examined in 151 consecutive, hospitalized drug abusers. Hepatitis B was diagnosed in 101 patients (66.8%), hepatitis A in 13 (8.6%) and non-A, non-B hepatitis in 35 (23.1%). Non-A, non-B hepatitis was significantly more prevalent among drug abusers than in an age-matched control population of non-drug abusers. Moreover, the mean duration of parenteral drug abuse was significantly lower among non-A, non-B cases than in patients with hepatitis A or B. These results suggest a wide circulation of both hepatitis B virus and non-A, non-B agent(s) among drug abusers in our area. These patients most likely represent a main reservoir of non A, non-B infection due to the high rate of chronicity reported for non-A, non-B hepatitis. PMID- 6293975 TI - Hematogenous Candida vertebral osteomyelitis treated with ketoconazole. AB - Candida vertebral osteomyelitis was diagnosed in a patient with systemic lupus erythematodes following X-ray evidence of osteomyelitis and the repeated culturing of Candida albicans from material obtained by needle biopsies from the third lumbar vertebra. The patient had been on glucocorticosteroids and parenteral nutrition six months previously. At that time, a yeast was cultured from the blood and the tip of the subclavian catheter which had been removed. After candida vertebral osteomyelitis was diagnosed, she was treated with ketoconazole for seven months. Recovery was impressive, as judged by the clinical and radiographic findings. At the time of writing this paper--12 months after the withdrawal of ketoconazole--the patient showed no signs of recurrence. PMID- 6293976 TI - The effect of N-formimidoyl thienamycin, ceftazidime, cefotiam, ceftriaxone and cefotaxime on non-fermentative Gram-negative rods, Aeromonas, Plesiomonas and Enterobacter agglomerans. AB - Thirty-one species (185 strains) of non-fermentative gram-negative rods (excluding Pseudomonas aeruginosa) as well as 45 strains of Aeromonas spp., 15 strains of Plesiomonas shigelloides and 68 strains of Enterobacter agglomerans were tested in microdilution procedures against N-formimidoyl thienamycin, ceftazidime, cefotiam, ceftriaxone and cefotaxime. N-formimidoyl thienamycin was the most effective drug as far as the spectrum of these bacterial groups and potency is concerned; ceftazidime was the second most effective agent. Ceftriaxone and cefotaxime were similar in their activity (against a smaller spectrum), while cefotiam showed little effect. There were occasional differences between MBC and MIC values which were most notable with ceftazidime, cefotiam, ceftriaxone and cefotaxime against E. agglomerans. PMID- 6293977 TI - Comparative beta-lactamase hydrolysis of and inhibition by 7-aminothiazolyl alpha methoxyimino cephalosporins. AB - Six 7-aminothiazolyl alpha-methoxyimino cephalosporins were found to be most variable in their ability to resist Type IV beta-lactamases (ceftizoxime, most stable and ceftriaxone, least stable), to inhibit Type I beta-lactamases (cefotaxime, the best inhibitor and desacetyl-cefotaxime, the least inhibition), to inhibit Enterobacteriaceae as measured by their minimum inhibitory concentrations (MICs), and in their published serum half-lives. The 3-position substituents appear to exert significant physical-chemical effects that influence the pharmacology, molecular enzyme stability and the antimicrobial activity of these compounds. However, we conclude that these differences are of minimal clinical consequence and that other more relevant factors for in vivo drug selection should be considered, including proven clinical efficacy and cost. PMID- 6293978 TI - Ceftriaxone: in vitro activity against 410 bacterial isolates compared with cefotaxime. AB - The in vitro activity of the two new cephalosporins, cefotaxime and ceftriaxone, against 410 bacterial isolates was compared using an agar dilution method. Both compounds were highly active against Enterobacteriaceae, including indole positive Proteus and Providencia; the great majority of the isolates were inhibited by 0.06 mg/l of either drug. Activity against Pseudomonas aeruginosa and Staphylococcus aureus was moderate, and enterococci were resistant. All Streptococcus pneumoniae, Streptococcus pyogenes and Haemophilus influenzae isolates were susceptible to 0.03 mg/l of either drug. The isolates belonging to the Bacteroides fragilis group were inhibited over a wide range of concentrations and some were highly resistant (MIC greater than or equal to 64 mg/l). There were no significant differences in the antibacterial activity of the two drugs against our isolates. Both drugs may be of potential use in the treatment of serious infections caused by Enterobacteriaceae; they may prove to be a useful alternative to the aminoglycosides. PMID- 6293979 TI - Complications of transfusion. PMID- 6293980 TI - Complement components, C1 activation and disease activity in SLE. AB - Laboratory parameters were studied in 8 systemic lupus erythematosus patients during periods of high and low disease activity, mainly as defined by clinical criteria. Renal manifestations were present in 6 patients 5 of which showed antibodies to native DNA. C-reactive protein was raised in 3 patients. Only 1 of these showed a superimposed bacterial infection. Markedly high concentrations of C1r-C1s-C1 inactivator cOmplexes (C1r-C1s-Cl IA) in the sera provided direct evidence of C1 activation independent of disease activity. During active disease. C1r-C1s-C1 IA were correlated with C1q binding immune complexes as measured by solid phase, but not by fluid phase assay. Immunochemical concentrations of C1q, C4 and C3 and functional C2 were decreased in active SLE, consistent with sequential activation of the classical pathway. Discrepancies were noted between functional and immunochemical assay for C2 but not for factor B. Although essentially within the normal range, the levels of C1s, C4 binding protein, C5 and properdin were lower during active than during inactive disease. The concentrations of the factors B, I and H did not suggest involvement of the alternative pathway. 1 exceptional patient showed low factor B, a relative decrease of factor I and the presence of Bb fragments in plasma during active SLE. Markedly high factor D values were found. This could partly be explained by reduced renal function. PMID- 6293981 TI - Release of eosinophil chemotactic leukotriene from neutrophils of patients with pseudo-allergic reactions. AB - Possible alterations in the release of eosinophil chemotactic leukotrienes (ECL) were studied with leukocytes from 14 patients with chronic urticaria due to pseudo-allergic reactions (PAR) and from 14 healthy volunteers. Patients tended to release less ECL, and inhibition of ECL release was more marked in the presence of cyclooxygenase and lipoxygenase inhibitors. The latter compounds did not cause ECL secretion by themselves. Increased release of lipoxygenase products of arachidonate metabolism from blood leukocytes is therefore an unlikely cause for PAR. PMID- 6293982 TI - Prolactin receptors in interstitial cells of testes from rats at different stages of development. AB - The developmental pattern of prolactin receptors was examined in intact interstitial cells isolated from testes of 19-day old foetuses (F19), 1 to 45-day immature (N1 to N45) and adult animals. The prolactin binding activity in F19 interstitial cells is low, being 27% of that found in cells of the adult. There is an abrupt 60% increase in prolactin binding immediately after birth (N1), followed by a slow, but gradual, increase for the next 34 and adulthood which follows an increment in serum prolactin concentrations after day 24. Equilibrium analysis revealed that the receptors In interstitial cells from N1, N34 and adults have similar affinity (Kd = 2.7 to 3.3 x 10(-10)M) for prolactin. However, cells from adults contain twice as many prolactin receptors per cell as that of immature animals. This increase in prolactin receptors may play a role in the modulation of interstitial (Leydig) cell function during sexual maturation. PMID- 6293983 TI - Effect of lead chelation therapy with EDTA in children on erythrocyte pyrimidine 5'-nucleotidase and with cytidine triphosphate levels. AB - Children with elevated whole blood lead levels were treated for 4 days with EDTA. Changes in whole blood lead concentrations, erythrocyte zinc protoporphyrin concentrations, red cell pyrimidine 5'-nucleotidase activities, and erythrocyte cytidine triphosphate (CTP) levels were measured during the course of the EDTA therapy. EDTA treatment decreased blood lead levels by approximately 50% after 4 days. An inverse relationship existed between blood lead levels and erythrocyte pyrimidine 5'-nucleotidase activity. Despite reactivation of pyrimidine 5' nucleotidase enzyme activity by EDTA, there was no decrease in erythrocyte cytidine triphosphate or in zinc protoporphyrin concentrations. The lack of change in these two parameters during EDTA treatment supports the concept that pyrimidine 5'-nucleotidase inhibition by lead in the reticulocyte cytidine phosphate accumulation in moderate lead poisoning may reflect chronic lead overburden analogous to prolonged elevation of erythrocyte zinc protoporphyrins. PMID- 6293984 TI - Bowenoid papulosis of the genitalia. PMID- 6293986 TI - Synthesis of human beta-endorphinyl-thiolglycine and its use for the preparation of affinity columns of beta-endorphin. AB - The peptide human beta-endorphinyl-thiolglycine (I) has been synthesized by the solid-phase method. The citraconyl derivative of peptide I was coupled to aminohexyl-Sepharose by reaction with silver nitrate/N-hydroxysuccinimide in water. The citraconyl groups were removed in aqueous acetic acid and the resulting resin was used in affinity chromatography for the purification of antisera to beta-lipotropin and beta-endorphin. PMID- 6293985 TI - Beta-endorphin: synthesis and properties of analogs with replacement of lysine residues by arginine. PMID- 6293987 TI - Chemical properties of water-soluble porphyrins 3. The reaction of superoxide radicals with some metalloporphyrins. AB - The catalytic efficiency of some water soluble metalloporphyrins upon the disproportionation of the superoxide radicals has been determined by observing the rate of decay of the radical at 254 nm. The Fe(III), Mn(III), Co(III), Ni(II), Cu(II) and Zn(II) derivatives of tetrakis-(4-N-methylpyridyl) porphyrin (TMPyP), tetra(4-N,N,N-trimethylanilinium) porphyrin (TAP), and tetra(4 sulphonatophenyl) porphyrin (TPPS4) were studied. The order of catalytic efficiency found was Fe(III)TMPyP much greater than Mn(III)TMPyP greater than Co(III)TMPyP approximately equal to Mn(TAP) greater than Fe(III)TPPS4. Other metalloporphyrins did not show catalytic activity. Reduction potentials of the catalytically active metalloporphyrins determined from cyclic voltamograms indicate the importance of having a second energetically accessible oxidation state available to the metal ion for catalysis. The catalytic action of the above metalloporphyrins is discussed in terms of inner and outer-sphere mechanisms. It is suggested that with the exception of Fe(III)TMPyP which reacts via an inner sphere mechanism, the others are governed by an outer-sphere mechanism. PMID- 6293988 TI - Antibody to acetylcholine receptors in human MG. PMID- 6293989 TI - Myasthenia gravis and acetylcholine receptor: autoimmunity and steroid effects. PMID- 6293990 TI - The immunopathology of myasthenia gravis. PMID- 6293991 TI - Experimental myasthenia gravis: a model of receptor disease. PMID- 6293992 TI - End-plate acetylcholinesterase deficiency associated with small nerve terminals and reduced acetylcholine release. A new syndrome. PMID- 6293993 TI - Treatment of schizophrenic patients in their homes through a visiting nurse- some issues in the nurse's training. PMID- 6293994 TI - Food borne infection with Clostridium perfringens type A. AB - Clostridium perfringens type A is one of the four most important bacterial agents causing food poisoning. Differential biochemical characterization appears to be important because of certain confounding species. Both the heat sensitive and resistant spore forming strains cause food poisoning. Ubiquitous nature, ability to contaminate carcass intravitally, thermal resistance of spores, relatively short generation time at high incubation temperature, survival in chilled and frozen meat and ability to overcome stomach acid barrier makes Cl. perfringens type A a common agent of food borne infection. Enteropathy is mediated through enterotoxin synthesised in vivo during sporulation. The enterotoxin is relatively insensitive to intestinal enzymes and is cytotoxic. The membrane bound transport enzyme, Na+K+-ATPase seems to be affected resulting into fluid accumulation in the ligated ileal loop of rabbit. Agglutination test using vegetative cell antigen is the commonly used epidemiological tool. New tools such as agglutination test with antispore serum, precipitin-in-gel test with anti enterotoxin serum, active and passive bacteriocin, bacteriophage and biotyping have been developed using standard strains and isolates of c. perfringens type A from foods, food poisoning and others. Except for bacteriophage typing, others appears to be useful alone or in combination, though these require to be tested in field investigations. PMID- 6293995 TI - Serological evidence of Japanese encephalitis virus activity in Bihar. AB - Serum samples from 404 pigs originating from eight districts of Bihar were examined for haemagglutination-inhibiting and complement fixing antibodies of Japanese Encephalitis, Dengue-2, West Nile and Chikungunya viruses. Japanese Encephalitis viral infection was found to be widely prevalent compared to other arboviruses tested. RElatively large number of pigs had antibodies to JEV only (i.e. monoconverters). High geometric mean titre to JEV was observed. These observations appear to be of epidemiologic significance to the 1978 epidemic of encephalitis in Bihar. Epidemic of encephalitis was reported in 1978 from several states of India including Bihar. The only evidence of the activity of Japanese Encephalitis in Bihar came from Khan and Banerjee who found neutralizing antibodies in ardeid birds from Asansol - Dhanbad regions. Evidences are presented in this paper on the prevalence of Japanese Encephalitis in eight districts including Dhanbad of Bihar. PMID- 6293996 TI - Intestinal loss of vitamin D3 metabolites in rats with cirrhosis of the liver. AB - The metabolism of cholecalciferol (vitamin D3) was studied in cirrhotic and in control rats that were either depleted of or supplemented with vitamin D3. The vitamin was supplemented by s.c. injections of radiolabeled cholecalciferol at doses which meat the daily nutritional requirements of rats. Excretion of vitamin D3 metabolites in the feces of the cirrhotic rats was greater than that in the control rats. Fecal excretion consisted mainly of vitamin D3 ester the unchanged vitamin D3 and 25-hydroxyvitamin D3. It is suggested that one of the causes for vitamin D3 depletion in cirrhosis of the liver is fecal loss. PMID- 6293997 TI - Inhibition of angiotensin-converting enzyme: therapeutic implications. PMID- 6293998 TI - Plasma opioid levels in post-traumatic chronic headache and trigeminal neuralgia: maintained response to acupuncture. PMID- 6293999 TI - A study of radiation parameters in an open-pit mine. AB - Radiation parameters associated with the open pit mining of a small (10,000 tonnes), but high grade (2%) uranium deposit at Nabarlek, N.T., have been investigated in detail. External radiation levels, radon emanation rates and radon daughter levels were measured systematically during the development of the mine, and are correlated with ore grade, properties of the host rock and atmospheric conditions. Significant radon daughter concentrations were observed only under stable atmospheric conditions, usually during the might and were invariably associated with thermal inversions. The mean cumulative exposure to radon daughters was estimated from the measured levels to be 0.065 Working Level Months for employees working in the pit for the entire four and a half months of mining. The mean cumulative external gamma ray exposure for the same employee group was measured using thermoluminescent dosimeters to be 2.3 mSv (230 mrem). For most other employees, however, exposures were much lower. Data on long lived radionuclides in dust and on particle size distribution are also presented. PMID- 6294000 TI - Paranasal sinus radiology, Part 2B: ethmoidal sinuses. AB - The clinician must have a high level of suspicion in order to detect ethmoidal sinus disease. Survey plain film radiographic examination may provide information about gross changes in ethmoidal sinus transparency and the sinus walls. Conventional tomographic study will often aid evaluation of the ethmoidal sinus walls which may be obscured by overlying structures on plain films. Computerized tomography adds a new dimension to ethmoidal sinus study. By revealing not only bone changes but soft-tissue abnormality as well, more complete understanding of the disease process is gained. Computerized tomography is particularly useful in evaluating orbital structural change due to ethmoidal sinus abnormality. PMID- 6294001 TI - [Malignant tumors of the salivary glands]. PMID- 6294002 TI - Homozygous alpha thalassemia/Hb G Philadelphia. AB - Microcytic red cells from a 70 year old Negro man with mild anemia contained only hemoglobin G-Philadelphia. Red cells from all of his children had low-normal MCV's, and contained 32-34 percent of the abnormal hemoglobin. Oxygen affinity of his blood and stability of his hemolysate were normal, suggesting that his mild anemia was not caused by the the abnormal hemoglobin. Restriction endonuclease analyses of DNA from the proband and his offspring showed that the alpha G Philadelphia globin gene exists in only one copy per chromosome. The new gene was probably created by an unequal cross-over which deleted an alpha globin coding sequence (derived from one or both alpha globin genes), as well as some or all of the DNA sequence between those genes. PMID- 6294003 TI - Alteration of hemoglobin function by two aliphatic amine buffers. PMID- 6294005 TI - Binding of bile salts to fibre-enriched wheat bran. AB - A new commercial product of fibre-enriched wheat bran was tested for its binding of bile salts in vitro. The wheat bran preparation was standardized and through enzymatic digestion of protein and starch contained 78 per cent fibre (w/w). Fibre-enriched wheat bran bound with high capacity both conjugated and unconjugated bile salts. Binding was saturable, reversible and showed no specificity towards tauro- or glycine-conjugated bile salts. Binding was rapid, dependent on pH, was enhanced by the presence of high salt concentrations and partially inhibited by 6 M urea. This indicated that binding was a combination of hydrophobic and hydrophilic interactions. PMID- 6294004 TI - Assessment of the metabolic effects of dietary carbohydrate and fibre by measuring urinary excretion of C-peptide. AB - An assessment of the metabolic effect of dietary carbohydrate on daily insulin secretion, reflected by the 24-h urine output of C-peptide, has been made. Urinary C-peptide excretion increased when the carbohydrate intake of 6 normal subjects was increased from 200 to 400 g. A high-fibre diet rich in beans and lentils caused a significant fall in urine C-peptide and a lowering of blood glucose values in both normal and diabetic subjects. This confirms the insulin sparing effect of leguminous foods. An increase of dietary fibre to 50 g mainly as cereal fibre did not significantly alter urine C-peptide excretion in normal subjects. Urinary C-peptide estimations could be useful for assessing the effect of different diets on daily insulin secretion. PMID- 6294006 TI - The cytochemical demonstration of NaK dependent adenosine triphosphatase at electrocyte level in Eigenmannia virescens (Gymnotidae). AB - ATPase activity (E.C. 3.6.1.3.) has been studied by electron microscopy with the help of several cytochemical techniques on Eigenmannia virescens electrocytes. Incubation was carried out with in two different media containing paranitrophenyl phosphate (p-NPP) or adenosine triphosphate (ATP) as substrate. With p-NPP the phosphate freed is captured at alkaline pH, either by strontium chloride or by lead citrate. With ATP the phosphate freed is captured at a pH close to neutrality by the lead nitrate. NaK ATPase activity was only demonstrated with the medium containing ATP; the positive results obtained with this technique were sensitive to ouabain. The enzyme is situated both on the membrane of the posterior face which is innervated and on that of the anterior face of the electrocytes. The cytoplasm of the anterior face is occupied by a strong concentration of tubules on whose membranes the enzyme is also present. The localisation of the enzyme on the tubules can explain biochemical results which indicate that 70% of the total NaK ATPase of the electrolytes is situated at the level of the anterior face. PMID- 6294007 TI - A cytochemical method for the demonstration of 5'-nucleotidase in mouse peritoneal macrophages, with cerium ions used as trapping agent. AB - A method was developed for the demonstration of 5'-nucleotidase in murine peritoneal resident macrophages. The cells are incubated cytochemically without agitation and cerium chloride is used as a trapping agent. Under these conditions, the great majority of the macrophages in the unstimulated peritoneal cavity show enzyme activity in the plasma membrane. In the presence of AMP-S (an AMP analogue inhibiting 5'-nucleotidase, as shown biochemically) there was a decrease in both the number of positive macrophages and the amount of reaction product on the plasma membranes. This indicates that the enzyme activity detected by our cytochemical procedure is attributable to 5'-nucleotidase. PMID- 6294008 TI - Can increased beta-endorphins explain the etiology of premenstrual syndrome? PMID- 6294009 TI - Beta-endorphins in fetal cord blood: correlation to maternal stress during parturition. PMID- 6294010 TI - How does increased cardiac output increase shunt in pulmonary edema? AB - In pulmonary edema, the relationship between cardiac output (QT) and shunt (QS/QT) may be due to a diffusion barrier for O2 transfer (incomplete alveolar capillary equilibration) or to redistribution of increased pulmonary blood flow toward edematous units. We compared transfer of O2 and multiple inert gases in the left (LLL) and right (RLL) lower lobes and in the whole lungs of eight dogs having oleic acid edema in LLL. When mean QT was increased from 3.0 to 5.5 l X min-1 during O2 ventilation, relative perfusion of LLL did not increase but QS/QT increased because LLL shunt increased from 56 to 78%. We conclude that increased pulmonary blood flow is not redistributed toward edematous regions, but we cannot exclude such redistribution within LLL and other slightly edematous lobes. In LLL, inert gas shunt and O2 shunt were not systematically different during O2 ventilation, and lobar venous PO2 measured during air ventilation was not different from that predicted by inert gas transfer. We conclude that diffusion limitation for O2 does not contribute to QS/QT or to the increase in QS/QT when QT increases. Conceivably, increased QT increased QS/QT by increasing edema or hematocrit in edematous regions. PMID- 6294011 TI - Helping nurses learn assertiveness. PMID- 6294012 TI - Futurism scenario: commencement address to the class of 2010. PMID- 6294013 TI - Utilizing a community data base system with community health nursing students. AB - The instrument to "go out and study the community" can be awesome and confusing to both students and nurses. The use of the data base management system as described in this paper makes the task a less formidable one. It also systematically classifies data about a community into a usable form so that nursing diagnoses can be identified appropriately. We strongly believe that the concept of community-as-client must become a part of the philosophy of every community health nurse. We see the data base management system as one method of incorporating this philosophy. PMID- 6294014 TI - How to conduct an assertion training course for nursing students: a step-by-step plan for instruction. PMID- 6294015 TI - Well elderly screening clinics: a community clinical experience in health assessment. PMID- 6294016 TI - Impact of a human sexuality workshop on the sexual attitudes & knowledge of nursing students. PMID- 6294017 TI - First Annual Scientific Meeting of Research in Nursing Education .... Abstracts. PMID- 6294018 TI - A teaching strategy for evaluating assertive behavior change. PMID- 6294019 TI - A comparison of lecture-discussion and self-study methods in nursing education. AB - Three groups of students in a Nurse Practitioner Program were subjected to a multiple-choice test based upon two articles from the Nurse Practitioner. Prior to testing, one group read and studied the articles for one hour with no discussion permitted. The other two groups did not read the articles but participated in a lecture-discussion on their contents. The testing followed each learning session. Overall, the results support strongly the hypothesis that the Lecture-Discussion method is superior to the Reading Only method. The fact that the Lecture-Discussion method utilized two different instructors suggests that this finding is not based on some unique personality characteristic of one particular faculty member. Naturally, these findings are limited to this particular subject matter with nurse practitioner students. It is possible that the Reading Only method would be equal to or superior to the Lecture-Discussion method with other topics or with other student bodies. In any case, the present results run counter to the current wave of enthusiasm for the Self-study method in nursing education. These results suggest that caution be employed in a wholesale shift to autotutorial results. As Hogan states: "Self-instructional materials are not replacements for . . . teacher-centered instruction. Instead they are supplements that encourage individual initiative, autonomy, and responsibility for students who can use and wish to use them." PMID- 6294020 TI - Art therapy: a learning experience for students in nursing. PMID- 6294021 TI - Development of a computer simulation to evaluate the clinical performance of nursing students. PMID- 6294022 TI - A self-directed clinical practicum. PMID- 6294023 TI - Assessment of the consumer's potential response to the nurse practitioner model. AB - A very positive atmosphere of acceptance for the nurse practitioner model exists in the general population. There is not complete acceptance for the NP role, and there remains a sizeable number of individuals who have not considered the issue, much less resolved it. Public relations, through mass media could significantly influence and alter consumer opinion, but no media campaign is so essential as the 90% rate of satisfaction reported by the 231 subjects who were pleased or very pleased with their prior nurse practitioner treatment. PMID- 6294024 TI - Teaching diploma students how to utilize the ANA Quality Assurance Model. PMID- 6294025 TI - Cell-mediated immunity: induction and expression of T-cell function. PMID- 6294026 TI - Ontogeny of the immune system. AB - Cellular and humoral aspects of the immune response develop sequentially in the fetus. Although there is evidence of immunologic activity to some antigens, many of the nonimmune effector systems are neither developed nor fully functional in the fetus. The lack of fully functional systems make the fetus a highly susceptible candidate for invading bacteria and viruses. Often, infection prior to immune competence may lead to abortion, malformation, and in some instances, viral persistence and immune tolerance. On the other hand, immune responses tend to elevate immunoglobulin values and cause specific antibody that may be used for diagnostic purposes. Little work has been done to appreciate fully the immunologic and nonimmune effector systems' role in normal development or during congenital infections. PMID- 6294027 TI - Respiratory tract immune response to microbial pathogens. AB - Effective resistance to respiratory tract infection depends principally on specific immunity on mucosal surfaces of the upper or lower respiratory tract. Respiratory tract immune response comprises antibody and cell-mediated systems and may be induced most readily by surface presentation of replicating agents but can result from parenteral or local presentation of highly immunogenic antigens. Upper and lower respiratory tract systems differ in immunologic competence, with the lungs having a greater inventory of protective mechanisms than the trachea or nose. Several effective vaccines have been developed for prevention or modification of respiratory tract diseases. PMID- 6294028 TI - Immune response to tumor cells in domestic animals. PMID- 6294029 TI - Theoretical and practical aspects of an immunization program for dogs and cats. PMID- 6294030 TI - Immunoprophylaxis in the horse. PMID- 6294031 TI - Theory and practice of immunoprophylaxis in cattle. PMID- 6294032 TI - Mechanisms of action by immunologic adjuvants. PMID- 6294033 TI - Candidiasis in captive cetaceans. AB - Disseminated Candida albicans infections were found or suspected in 4 captive cetaceans. Ketoconazole at a dosage of 2.5 mg/kg, BID, administered orally for 18 days, followed by 8 biweekly oral doses of levamisole hydrochloride at the rate of 9 mg/kg, resulted in regression of clinical signs of candidiasis in an adult male Atlantic bottlenose dolphin (Tursiops truncatus). A higher dosage of ketoconazole (6 mg/kg, BID) was effective in eliminating the shedding of C albicans from an adult male belukha whale (Delphinapterus leucas). A juvenile female harbor porpoise (Phocoena phocoena) treated with nystatin died with disseminated candidiasis, as did a juvenile male longfinned pilot whale (Globicephala melaena) treated with nystatin and levamisole. Three other adult bottlenose dolphins, a juvenile female belukha whale, and a female Commerson's dolphin (Cephalorhynchus commersonii) kept in the same water system never had evidence of candidiasis. A 5th bottlenose dolphin (an adult female) was culture positive on 1 occasion, but never had signs of the disease. PMID- 6294034 TI - Herpesvirus-like infection in map turtles. AB - Two map turtles housed in an aquarium with a group of other map turtles became lethargic and anorectic, and subcutaneous edema developed prior to their deaths. Gross postmortem findings included subcutaneous edema and swollen, diffusely pale livers. Histologic examination of the livers revealed multifocal to diffuse areas of necrosis, with intranuclear eosinophilic inclusions in hepatocytes. Intranuclear and intracytoplasmic aggregates of particles morphologically resembling herpesvirus were identified in degenerating hepatocytes by electron microscopy. PMID- 6294035 TI - Papilloma-like virus infection in Bolivian side-neck turtles. AB - Five Bolivian side-neck turtles had multifocal small, round to confluent, white skin lesions distributed over the head. Several gram-negative microorganisms were isolated from the lesions. Light microscopy revealed hyperkeratosis and hyperplasia of the epidermis. Ultrastructural evaluation demonstrated crystalline aggregates of virus particles within nuclei of cells in the stratum granulosum and free within the stratum corneum. On the basis of size, location, arrangement, and tissue affected, the particles resembled papillomaviruses. PMID- 6294036 TI - A treatment regimen for candidiasis in an Atlantic bottlenose dolphin. PMID- 6294037 TI - Oral therapy for nasal cryptococcosis in a cheetah. PMID- 6294038 TI - Inapparent bluetongue in free-ranging white-tailed deer. PMID- 6294039 TI - Fatal Herpesvirus hominis encephalitis in a white-handed gibbon. PMID- 6294040 TI - The inactivation of Bacillus cereus 569/H bera-lactamase by 6-beta (trifluoromethane-sulfonyl)amidopenicillanic acid sulfone: pH dependence and stoichiometry. PMID- 6294041 TI - Total chemical structure of streptothricin. PMID- 6294042 TI - Residues in the fat of ewes grazing on soil contaminated with halogenated hydrocarbons. AB - Three halogenated hydrocarbon compounds were applied to the surface of each of two .57 ha bluegrass plots in October. Each plot was stocked with 10 mature ewes 6 mo after application. Supplemental feed was not offered. Five ewes remained on the plots for 180 d, whereas the other five were removed and replaced at 60-d intervals. Average soil residues for the period of grazing were 13.3, 3.8, 29.3 and 32.8 mg/m2 HCB (hexachlorobenzene), DDE [1,1-dichloro-2,2-bi(p chlorophenyl)ethylene], DDT [1,1,1-trichloro-2,2-bis-(p-chlorophenyl)ethane] and PBB (polybrominated biphenyls), respectively, for plot 1 and 16.5 and 48.0 mg/m2 DDE and PBB, respectively, for plot 2. Average concentrations of residue in body fat of the five ewes grazing for 180 d were .37, .30 and .30 micrograms/g HCB, DDE and PBB, respectively, for plot 1 and 2.41 and .79 micrograms/g DDE and PBB, respectively, for plot 2. Average residue concentrations in ewes that grazed 60-d subperiods were nearly as great, which indicated that steady state residue levels were reached in less than 180 d. A second trial and was conducted 3 yr later after plowing and reseeding the plots, but only PBB was measured. Polybrominated biphenyl was distributed throughout the top 16 cm of soil but the quantity present had not changed appreciably from the first trial. After seven ewes/plot grazed 136 d, residues were detected in only one of the seven on plot 1, but were detected in all seven on plot 2 with an average concentration of only .032 micrograms/g. It is concluded that concentration of residue in body fat of ewes depended on the concentration of chemical at the soil surface and the amount of soil ingested. PMID- 6294043 TI - In-vitro activity of cefotiam against bacteria of clinical interest. PMID- 6294044 TI - Inoculum size effect on the MIC of cefoperazone, moxalactam, cefotaxime, cefoxitin and cephalothin for 118 strains of Haemophilus influenzae including 'tolerant' micro-organisms. PMID- 6294045 TI - Combination of sulbactam pivoxyl and bacampicillin in acute exacerbations of chronic bronchitis. PMID- 6294046 TI - Tn1721-encoded tetracycline resistance: mapping of structural and regulatory genes mediating resistance. AB - The genes encoding inducible tetracycline resistance in Tn1721 were located in a 2.1-kilobase portion of the transposon. Using deletions and insertions, we mapped and characterized two tet genes by their mutant phenotypes. Two tetracycline inducible polypeptides synthesized in minicells were assigned to the tet genes. The polarity of the tet genes was determined by employing a deletion and a gene fusion which altered the carboxy termini of the polypeptides. The gene responsible for resistance (tetA) encompasses 1,250 base pairs and encodes a membrane-bound protein with an apparent molecular weight of 34,000. The second gene (tetR) encompasses at least 650 base pairs and encodes a soluble 26,000 dalton protein, identified by complementation analysis as the repressor. The two adjacent genes have opposite transcriptional polarity, suggesting that the sites controlling their expression are located in the intercistronic region between tetA and tetR. PMID- 6294047 TI - Mechanism of CRP-mediated cya suppression in Escherichia coli. AB - Escherichia coli strain NCR30 contains a cya lesion and a second-site cya suppressor mutation that lies in the crp gene. NCR30 shows a pleiotropic phenotypic reversion to the wild-type state in expressing many operons that require the cyclic AMP (cAMP)-cAMP receptor protein (CRP) complex for positive control. In vivo beta-galactosidase synthesis in NCR30 was sensitive to glucose mediated repression, which was relieved not only by cAMP but also by cyclic GMP and cyclic CMP. The CRP isolated from NCR30 differed from the protein isolated from wild-type E. coli in many respects. The mutant protein bound cAMP with four to five times greater affinity than wild-type CRP. Protease digestion studies indicated that native NCR30 CRP exists in the cAMP-CRP complex-like conformation. The protein conferred a degree of cAMP independence on the in vitro synthesis of beta-galactosidase. In addition, the inherent positive control activity of the mutant protein in vitro was enhanced by those nucleotides that stimulate in vivo beta-galactosidase synthesis in NCR30. The results of this study supported the conclusion that the crp allele of NCR30 codes for a protein having altered effector specificity yet capable of promoting positive control over catabolite sensitive operons in the absence of an effector molecule. PMID- 6294048 TI - Porin channels in Escherichia coli: studies with beta-lactams in intact cells. AB - Wild-type Escherichia coli K-12 produces two porins, OmpF (protein 1a) and OmpC (protein 1b). In mutants deficient in both of these "normal" porins, secondary mutants that produce a "new" porin, protein PhoE (protein E), are selected for. We determined the properties of the channels produced by each of these porins by measuring the rates of diffusion of various cephalosporins through the outer membrane in strains producing only one porin species. We found that all porin channels retarded the diffusion of more hydrophobic cephalosporins and that with monoanionic cephalosporins a 10-fold increase in the octanol-water partition coefficient of the solute produced a 5- to 6-fold decrease in the rate of penetration. Electrical charges of the solutes had different effects on different channels. Thus, with the normal porins (i.e., OmpF and OmpC proteins) additional negative charge drastically reduced the penetration rate through the channels, whereas additional positive charge significantly accelerated the penetration. In contrast, diffusion through the PhoE channel was unaffected by the presence of an additional negative charge. We hypothesize that the relative exclusion of hydrophobic and negatively charged solutes by normal porin channels is of ecological advantage to E. coli, which must exclude hydrophobic and anionic bile salts in its natural habitat. The properties of the PhoE porin are also consistent with the recent finding (M. Argast and W. Boos, J. Bacteriol. 143:142 150, 1980; J. Tommassen and B. Lugtenberg, J. Bacteriol. 143:151-157, 1980) that its biosynthesis is derepressed by phosphate starvation; the channel may thus act as an emergency pore primarily for the uptake of phosphate and phosphorylated compounds. PMID- 6294050 TI - Pseudomonas aeruginosa outer membrane permeability: isolation of a porin protein F-deficient mutant. AB - A mutant of Pseudomonas aeruginosa severely deficient in outer membrane protein F levels was isolated by screening heavily mutagenized strains for membrane protein alterations on sodium dodecyl sulphate-polyacrylamide gel electrophoresis. To provide a basis for phenotypic comparison, three independent spontaneous revertants with normal protein F levels were isolated. Neither the protein F deficient mutant nor its revertants had gross surface alterations as judged by their sensitivities to 31 phages with diverse receptors and their low degrees of leakage of periplasmic beta-lactamase into the supernatant. Outer membrane permeability was measured in whole cells by examining the rates of hydrolysis of a chromogenic beta-lactam, nitrocefin, by periplasmic RP1-encoded beta-lactamase. It was found that the outer membrane permeabilities of wild-type and protein F revertant strains were similar, but low when compared with those of Escherichia coli and an antibiotic-supersusceptible mutant Z61 of P. aeruginosa. The loss of protein F caused a further significant decrease in outer membrane permeability. The results suggest that protein F is a pore-forming protein in vivo and that only a small proportion, as few as 1 in 400, of the protein F molecules form active functional channels in vivo. PMID- 6294049 TI - Porin channels in Escherichia coli: studies with liposomes reconstituted from purified proteins. AB - Rates of diffusion of uncharged and charged solute molecules through porin channels were determined by using liposomes reconstituted from egg phosphatidylcholine and purified Escherichia coli porins OmpF (protein 1a), OmpC (protein 1b), and PhoE (protein E). All three porin proteins appeared to produce channels of similar size, although the OmpF channel appeared to be 7 to 9% larger than the OmpC and PhoE channels in an equivalent radius. Hydrophobicity of the solute retarded the penetration through all three channels in a similar manner. The presence of one negative charge on the solute resulted in about a threefold reduction in penetration rates through OmpF and OmpC channels, whereas it produced two- to tenfold acceleration of diffusion through the PhoE channel. The addition of the second negatively charged group to the solutes decreased the diffusion rates through OmpF and OmpC channels further, whereas diffusion through the PhoE channel was not affected much. These results suggest that PhoE specializes in the uptake of negatively charged solutes. At the present level of resolution, no sign of true solute specificity was found in OmpF and OmpC channels; peptides, for example, diffused through both of these channels at rates expected from their molecular size, hydrophobicity, and charge. However, the OmpF porin channel allowed influx of more solute molecules per unit time than did the equivalent weight of the OmpC porin when the flux was driven by a concentration gradient of the same size. This apparent difference in "efficiency" became more pronounced with larger solutes, and it is likely to be the consequence of the difference in the sizes of OmpF and OmpC channels. PMID- 6294051 TI - Transposon insertion and subsequent donor formation promoted by Tn501 in Bordetella pertussis. AB - The mercuric chloride resistance transposon, Tn501, was introduced into Bordetella pertussis by using the chimeric plasmid pUW942, which is unable to replicate in this species. Tn501 insertions which conferred a thiamine requirement were the predominant insertion class. In many cases, the mercuric chloride-resistant transconjugants were also resistant to the other plasmid markers, but failure to detect plasmid DNA in these isolates indicated that integration of the entire plasmid into the chromosome had occurred. One such insertion was further characterized. Southern hybridization with a Tn501-specific probe indicated that chromosomal DNA from one strain containing the integrated plasmid had two copies of Tn501 and an intervening copy of the plasmid associated with the chromosome. The presence of the plasmid was unstable, and derivatives which had lost all of the plasmid markers except mercuric chloride resistance were obtained. These strains had a single copy of Tn501 and had lost all of the rest of the plasmid-specific sequences. Strains containing the plasmid in the integrated state could act as genetic donors and mobilize chromosomal genes. PMID- 6294052 TI - Isolation of Saccharomyces cerevisiae TRP3. AB - Several plasmids, isolated from two plasmid pools, complemented a Saccharomyces cerevisiae trp3 mutant with defective indole-3-glycerol-phosphate synthase activity. Restriction mapping indicated that a 1.2-kilobase StuI segment was common to all complementing plasmids. Southern blot hybridization established that a cloned 5.2-kilobase BamHI fragment was derived intact from chromosomal DNA. A yeast trp3 mutant transformed with trp3-complementing plasmids contained approximately 40-fold elevated indole-3-glycerol-phosphate synthase activity. These plasmids also complemented an Escherichia coli trpC mutant, and transformants exhibited enzyme activity. Yeast trp3 is therefore associated with a 1.2-kilobase StuI DNA segment. PMID- 6294053 TI - Effect of starvation on cytoplasmic pH, proton motive force, and viability of an acidophilic bacterium, Thiobacillus acidophilus. AB - The question of whether Thiobacillus acidophilus maintains its cytoplasmic pH at values close to neutrality by active or passive means was explored by subjecting the organism to long-term starvation (up to 22 days). Starving cells maintained a delta pH of 2 to 3 U throughout starvation, although cellular poly-beta hydroxybutyric acid and ATP, the proton motive force, and culture viability were low or not detectable after 200 h. Cells exposed to azide or azide plus N,N' dicyclohexylcarbodiimide immediately exhibited characteristics of cells starved for more than 200 h. Thus, a large delta pH in T. acidophilus was maintained in the absence of ATP, ATPase activity, respiration, significant levels of proton motive force, and cell viability and was therefore not dependent on chemiosmotic ionic pumping. The transition from a metabolically active to an inactive state was accompanied by a large increase in the positive membrane potential, which nearly completely compensated for the delta pH in the inactive cells. The longevity of the acidophile during starvation was comparable to that reported previously for neutrophiles, and the loss of viability occurred not because of the acidification of the cytoplasm but apparently because of energy depletion. PMID- 6294054 TI - New class of mutations in Escherichia coli (uup) that affect precise excision of insertion elements and bacteriophage Mu growth. AB - We have used a papillation screening technique to isolate mutations that increase the precise excision of insertion elements. The three mutations isolated stimulated precise excision of Tn5, Tn10, and the IS elements. They had a large, 20- to 600-fold, effect on excision of Tn5 at various chromosomal sites. The varied stimulation for different Tn5 insertions showed that the mutations altered the relationship between a precise excision activity and the chromosomal sequence flanking an inserted Tn5. A much smaller stimulation was observed for insertions on the plasmid F'128. The stimulation was recA independent. The mutations also reduced the rate of production of bacteriophage Mu progeny. The mutations were mapped by two- and three-factor crosses with closely linked Tn10 insertions. They defined the uup locus, located at 21.3 min on the Escherichia coli map, next to pyrD. PMID- 6294055 TI - Mapping of chromosomal IS5 elements that mediate type II F-prime plasmid excision in Escherichia coli K-12. AB - Three IS5 elements were mapped in overlapping chromosomal segments on a series of F-prime plasmids by restriction analysis and hybridization. IS5A was located clockwise of proA near 6 min, IS5B was located clockwise of purE near 12 min, and IS5C was tentatively located near 14 min on the Escherichia coli K-12 map. The physical structures of nine type II F-prime plasmids that contain chromosomal DNA from this region indicated that these plasmids were excised from the chromosome by recombination between pairs of IS5 elements. PMID- 6294056 TI - Characterization of the mgl operon of Escherichia coli by transposon mutagenesis and molecular cloning. AB - We used transposon insertion mutagenesis, molecular cloning, and a novel procedure for in vitro construction of polar and nonpolar insertion mutations to characterize the genetic organization and gene products of the beta methylgalactoside (Mgl) transport system, which utilizes the galactose-binding protein. The data indicate that the mgl operon contained three genes, which were transcribed in the order mglB, mglA, and mglC. The first gene coded for the 31,000 Mr galactose-binding protein, which was synthesized as a 3,000-dalton larger precursor form. The mglA product was a 50,000 Mr protein which was tightly associated with the membrane, and the mglC product was a 38,000 Mr protein which was apparently loosely associated with the membrane and was probably located on the internal face of the cytoplasmic membrane. Identification of gene products was facilitated by in vitro insertion of a fragment of Tn5 containing the gene conferring kanamycin resistance into a restriction site in the operon. The fragment proved to have a polar effect on the expression of promoter-distal genes only when inserted in one of the two possible orientations. The three identified gene products were necessary and apparently sufficient for transport activity, but only the binding protein was required for chemotaxis towards galactose. The transport system appeared to contain the minimum number of components for a binding protein-related system: a periplasmic recognition component, a transmembrane protein, and a peripheral membrane protein that may be involved in energy linkage. PMID- 6294057 TI - Escherichia coli mutants defective in the uncH gene. AB - Plasmids carrying cloned segments of the unc operon of Escherichia coli have been used in genetic complementation analyses to identify three independent mutants defective in the uncH gene, which codes for the delta subunit of the ATP synthetase. Mutations in other unc genes have also been mapped by this technique. ATPase activity was present in extracts of the uncH mutants, but the enzyme was not as tightly bound to the membrane as it was in the parental strain. ATP dependent membrane energization was absent in membranes isolated from the uncH mutants and could not be restored by adding normal F1 ATPase from the wild-type strain. F1 ATPase prepared from uncH mutants could not restore ATP-dependent membrane energization when added to wild-type membranes depleted of F1. Membranes of the uncH mutants were not rendered proton permeable as a result of washing with low-ionic-strength buffer. PMID- 6294058 TI - Integration of R91-5::Tn501 into the Pseudomonas putida PPN chromosome and genetic circularity of the chromosomal map. AB - Derivatives of the Pseudomonas aeruginosa plasmid R91-5, loaded with the transposon Tn501, were transferred to P. putida PPN. Over 90% of exconjugants, which arose at a frequency of ca. 10(-6) per donor cell, exhibited high-frequency (greater than 10(-2) per donor cell) polarized transfer of chromosomal markers. In one instance it was demonstrated by transduction that the plasmid had been inserted into a gene required for serine biosynthesis. The integrated nature of the plasmid in this and other P. putida (R91-5::Tn501) derivatives was supported by the failure to detect covalently closed circular DNA in these strains. The transfer origins of six different Hfr donors have been characterized genetically, and time-of-entry kinetics obtained from interrupted matings have enabled the construction of a circular genetic map 103 min in length and containing 35 markers. The genetic map of P. putida PPN shows significant differences in marker order to that of P. aeruginosa PAO. PMID- 6294059 TI - Genetic homology between independently isolated chlorobenzoate-degradative plasmids. AB - Two chlorobenzoate-degradative plasmids were studied by the hybridization of the restriction endonuclease-generated fragments of one plasmid after transfer to a nitrocellulose filter with nick-translated radioactive DNA of the other plasmid as a probe. Two strains harboring the 3-chlorobenzoic acid-degradative plasmids were isolated in two different parts of the world at two different times. The plasmids are now found to be closely related to each other by hybridization studies. The chlorobenzoate-degradative plasmid from Pseudomonas sp. strain B13 (termed pB13) has a 6-kilobase deletion but otherwise is homologous with previously described plasmid pAC25. PMID- 6294060 TI - Proton pump coupled to cytochrome c oxidase in the cyanobacterium Anacystis nidulans. AB - Intact spheroplasts of the cyanobacterium Anacystis nidulans were found to oxidize various exogenous c-type cytochromes with concomitant proton extrusion. In the coupled state, H+/e stoichiometries close to 1 were measured, regardless of absolute reaction rates. It is concluded that the proton translocation observed is an intrinsic property of the cytoplasmic membrane-bound cytochrome c oxidase of A. nidulans. PMID- 6294061 TI - Copy number mutations (Cop-) of the plasmid containing the replication origin (oriC) of the Escherichia coli chromosome: lethal effect of the cop region cloned onto a high-copy-number vector on host cells. AB - High-copy-number mutants were isolated from an oriC plasmid. They carried insertion mutations within a region (about 470 base pairs) near the uncB gene. When a segment containing this region was cloned onto a high-copy-number plasmid, such a plasmid could be maintained as an intact form only when it was present in a lower copy number. PMID- 6294062 TI - Absence of significant light-induced changes in cAMP levels in sporangiophores of Phycomyces blakesleeanus. AB - We were unable to find transient changes in the amount of cAMP or cGMP that had been proposed to mediate the light-growth response in sporangiophores of Phycomyces blakesleeanus. PMID- 6294063 TI - Activation and inactivation of methanol: 2-mercaptoethanesulfonic acid methyltransferase from Methanosarcina barkeri. AB - Methanol is converted to methane by crude extracts of Methanosarcina barkeri. The first reaction involved in this process, is catalyzed by methanol:2 mercaptoethanesulfonic acid methyltransferase (EC 2.1.1.-). The methyltransferase has an optimum at pH 6.5 and is not inhibited by 2-bromoethanesulfonic acid. Pyridoxal-5'-phosphate acts as an inhibitor (Ki = 0.30 mM). The methyltransferase was tested in the presence of 2-bromoethanesulfonic acid, which inhibits the conversion of 2-(methylthio)ethanesulfonic acid to methane. The reaction is subject to activation and inactivation. Inactivation is brought about by the presence of oxygen, flavin mononucleotide, flavin adenine dinucleotide, and 2 (methylthio)ethanesulfonic acid, the product of the reaction. Activation of the system requires the presence of ATP and Mg2+ and of hydrogen. Hydrogen can be replaced by enzymatic systems, such as pyruvate dehydrogenase, which deliver free hydrogen. PMID- 6294064 TI - Plasmid linkage of the D-tagatose 6-phosphate pathway in Streptococcus lactis: effect on lactose and galactose metabolism. AB - The three enzymes of the D-tagatose 6-phosphate pathway (galactose 6-phosphate isomerase, D-tagatose 6-phosphate kinase, and tagatose 1,6-diphosphate aldolase) were absent in lactose-negative (Lac-) derivatives of Streptococcus lactis C10, H1, and 133 grown on galactose. The lactose phosphoenolpyruvate-dependent phosphotransferase system and phospho-beta-galactosidase activities were also absent in Lac- derivatives of strains H1 and 133 and were low (possibly absent) in C10 Lac-. In all three Lac- derivatives, low galactose phosphotransferase system activity was found. On galactose, Lac- derivatives grew more slowly (presumably using the Leloir pathway) than the wild-type strains and accumulated high intracellular concentrations of galactose 6-phosphate (up to 49 mM); no intracellular tagatose 1,6-diphosphate was detected. The data suggest that the Lac phenotype is plasmid linked in the three strains studied, with the evidence being more substantial for strain H1. A Lac- derivative of H1 contained a single plasmid (33 megadaltons) which was absent from the Lac- mutant. We suggest that the genes linked to the lactose plasmid in S. lactis are more numerous than previously envisaged, coding for all of the enzymes involved in lactose metabolism from initial transport to the formation of triose phosphates via the D tagatose 6-phosphate pathway. PMID- 6294065 TI - Some characteristics of hydrogen- and alkylhydroperoxides metabolizing systems in cardiac tissue. AB - The effect of hydroperoxides on the cardiac tissue was studied by using hemoglobin-free perfused rat heart. Ethylhydroperoxide was degraded mainly through the glutathione peroxidase system of the heart at a maximal rate of about 1.2 mumol/min per g wet wt. When ethylhydroperoxide infused was not degraded completely, the hydroperoxide concentration in the effluent perfusate paralleled the formation of ferrylmyoglobin in the heart. The infusion of ethylhydroperoxide caused release of oxidized glutathione into the effluent perfusate as a result of the enhancement of the cytosolic glutathione peroxidase reaction. The leakage of oxidized glutathione reached the maximal rate of 3.5 nmol/min per g wet wt with the infusion of 175 microM ethylhydroperoxide. At hydroperoxide concentrations above 150 microM, oxidations of pyridine nucleotides and of cytochrome a + a3 occurred, probably through a stimulation of the mitochondrial glutathione peroxidase reaction, and resulted in sudden failure of the heart function. The infusion of t-butyl- and cumene-hydroperoxides, which are unable to react with myoglobin, also caused the oxidations of pyridine nucleotides and cytochrome a + a3, the inhibition of oxygen consumption and the failure of heart function. The results indicate that the cardiac toxicity of hydroperoxides is due mainly to their effect on mitochondrial metabolism. PMID- 6294066 TI - Purification and characterization of a protein from Escherichia coli which forms complexes with superhelical and single-stranded DNAs. AB - From the cells of an Escherichia coli K-12 strain, a 22,000-dalton protein which has an affinity for the superhelical DNA molecule was purified to apparent homogeneity by monitoring the DNA-binding activity using the filter binding assay. In the sedimentation analysis of the DNA-protein complex, the protein has an affinity for the superhelical or single-stranded DNA molecule but neither for the open-circular nor for the linear DNA molecule. The amino acid composition of the protein resembled those of the other prokaryotic histone-like proteins and also to eukaryotic histones H2A and H2B. The protein precipitated upon heating, which is in contrast to the heat-stable feature of the other histone-like proteins. Furthermore, DNA and RNA syntheses in vitro were not affected by the presence of the protein. In view of these characteristics, this protein may play a role in maintaining the bacterial nucleoid structure. PMID- 6294067 TI - Inactivation of ATP-dependent deoxyribonuclease of Micrococcus luteus by 2,3 butanedione. AB - ATP-dependent deoxyribonuclease from Micrococcus luteus was purified to near homogeneity by a procedure involving gentle cell lysis, ammonium sulfate fractionation, TEAE-cellulose chromatography, Sephadex G-150 gel filtration and DNA-cellulose chromatography. Treatment of the enzyme with 2,3-butanedione, which binds specifically to arginyl residues, caused rapid loss of enzyme activities and the effect was enhanced by borate ion. The reaction obeyed first order kinetics with respect to the butanedione concentration, indicating that at least one functional arginyl residue is involved in the inactivation reaction. The enzyme was protected from inactivation by the presence of a low concentration of ATP, but not of ADP, AMP or adenosine. These results indicate that ATP-dependent deoxyribonuclease of Micrococcus luteus has functional arginyl residue(s) at an ATP-binding site. PMID- 6294068 TI - The conventional and saturation transfer electron paramagnetic resonance of spin labeled myosin subfragment-1 in the presence of F-actin and nucleotides. AB - SH-1 thiol of S-1 was modified with N-(1-oxyl-2,2,6,6-tetramethyl-4-piperidinyl) iodoacetoamide spin label (IASL). The extent of dissociation, alpha, of spin labeled myosin subfragment-1 (IASL-S-1) from acto-IASL-S-1 by a nucleotide was measured by an ultracentrifugal separation method, a light-scattering method, and a saturation transfer EPR method. The alpha values obtained by these three methods were the same within the limits of the experimental errors. The dependence of alpha on the concentrations of AMPPNP, [S], and F-actin, [A], could be described by the equation: alpha-1 = 1 + (1 + Ks/[S])[A]/KA. The Ks and KA values were 0.65-1.2 mM and 1.7-2.7 mg/ml, respectively, in 0.5 M KCl and 4 mM MgCl2 at pH 7.0 and 20 degrees C. The height of the weakly immobilized peak of the conventional EPR spectrum of IASL-S-1, W, increased linearly with increase in the ATP or AMPPNP concentration, and became saturated at 1 mol nucleotide/mol IASL-S-1. No change in W was observed upon the binding of IASL-S-1 with F-actin. The dependence of the extent of change in W, delta W, on [A] and [S] was given by delta W-1 = 1 + Ks/[S], where Ks = Ks/(1 + KA/[A]). This finding indicates that the delta W value is proportional to the amount of a nucleotide bound to IASL-S-1 and independent of the binding of F-actin to IASL-S-1. PMID- 6294069 TI - Ca2+-induced Ca2+ release from fragmented sarcoplasmic reticulum: a comparison with skinned muscle fiber studies. AB - Uptake and release of Ca2+ in heavy and light fractions of fragmented sarcoplasmic reticulum (FSR) isolated from frog and rabbit skeletal muscle was studied under conditions similar to those employed in skinned muscle fiber experiments, where ATP and Mg2+ concentrations were considered to be physiological and free Ca2+ concentration was kept constant during the Ca2+ uptake and release. Ca2+ level in FSR monotonously approached a steady state level which depended only on the final experimental conditions. Heavy fractions, but not light fractions, exhibited characteristics similar to those of Ca2+ induced Ca2+ release reported in skinned fiber studies: i) the rate and steady state level of Ca2+ uptake increased with increase in free Ca2+ concentration in the reaction medium up to 10(-6) M. With further increase in free Ca2+ concentration, the steady state level of Ca2+ taken up decreased while the Ca2+ uptake rate increased. ii) The steady state Ca2+ level was decreased by caffeine but increased by procaine or ruthenium red. Parallel measurement of Ca2+-ATPase activity clearly showed that these drugs modify the Ca2+ efflux but hardly affect the Ca2+-pump activity. It was concluded that the Ca2+-induced Ca2+ release mechanism was in operation at as low as 10(-6) M free Ca2+ concentration. Treatment of FSR with 0.6 M KCl did not have any significant effect. PMID- 6294070 TI - Acridine orange as a fluorescent probe for lysosomal proton pump. AB - Acridine orange was found to accumulate in pure lysosomal particles (tritosomes) in vitro, and the quenching of its fluorescence correlated well with the delta pH (inside acid) across the lysosomal membrane. Use of this dye showed that Mg-ATP caused lysosomal acidification. This acidification was sensitive to N,N' dicyclohexylcarbodiimide, N-ethylmaleimide, and azide, but not to oligomycin, ouabain or vanadate. These results supported the idea of the existence of a lysosomal H+-pump, suggested in a previous paper (Ohkuma et al. (1982) Proc. Natl. Acad. Sci. U.S. 79, 2758-2762). PMID- 6294071 TI - Peptide mapping of multiple forms of cyclic nucleotide phosphodiesterase. AB - Purified multiple forms of 3':5'-cyclic-nucleotide phosphodiesterase (EC 3.1.4.17) were analyzed using two-dimensional tryptic pep]tide mapping of radioiodinated peptides. Comparisons of peptide maps of rat liver insulin sensitive phosphodiesterase (PDE) with rat brain calmodulin-sensitive PDE suggest that some peptides co-migrate (31-43% co-migration). However, except for a single peptide, bovine retinal rod outer segment PDE, peptide maps appear unrelated to the other two forms (7-12% co-migration). In contrast, peptide maps of a 36,000 dalton proteolysis product of calmodulin-sensitive PDE are highly related to the peptide maps of a rat brain calmodulin-sensitive holoenzyme (73% co-migration). These results suggest that the multiple PDE forms are distinct molecular entities. PMID- 6294072 TI - Evidence that changes in platelet cyclic AMP levels regulate the fibrinogen receptor on human platelets. AB - Fibrinogen binds to human platelets after specific receptor sites are exposed by thrombin, ADP, epinephrine, and other stimuli. Since prostaglandin I2 (PGI2), a potent activator of platelet adenylate cyclase, prevents mobilization of the fibrinogen receptor by aggregating agents, we investigated the relationship between platelet cAMP levels and fibrinogen receptor status in thrombin stimulated human platelets. A dose-dependent rise in platelet cAMP in response to two adenylate cyclase agonists, PGI2 and forskolin, correlated with progressive inhibition of fibrinogen binding. Moreover, the receptor inhibition produced by either agonist was sustained up to 2 h and was associated with a persistent increase in cAMP levels. The phosphodiesterase inhibitor, 1-methyl-3 isobutylxanthine, in the presence of a subthreshold concentration of PGI2 also raised cAMP and inhibited fibrinogen binding. In contrast, the effects of PGI2 on both cAMP and fibrinogen binding were markedly attenuated by 9-(tetrahydro-2 furyl) adenine, an adenylate cyclase inhibitor. These results indicate that the inhibition of fibrinogen binding by PgI2 is linked to its effect on cAMP levels and suggest that elevation of platelet cAMP levels from any cause prevents exposure of the fibrinogen receptor. PMID- 6294073 TI - Evidence for nickel and a three-iron center in the hydrogenase of Desulfovibrio desulfuricans. AB - Hydrogenase from Desulfovibrio desulfuricans (ATCC No. 27774) grown in unenriched and in enriched 61Ni and 57Fe media has been purified to apparent homogeneity. Two fractions of enzymes with hydrogenase activity were separated and were termed hydrogenase I and hydrogenase II. they were shown to have similar molecular weights (77,600 for hydrogenase I and 75,500 for hydrogenase II), to be composed of two polypeptide chains, and to contain Ni and non-heme iron. Because of its higher specific activity (152 versus 97) hydrogenase II was selected for EPR and Mossbauer studies. As isolated, hydrogenase II exhibits an "isotropic" EPR signal at g = 2.02 and a rhombic EPR signal at g = 2.3, 2.2, and 2.0. Isotopic substitution of 61Ni proves that the rhombic signal is due to Ni. Combining the Mossbauer and EPR data, the isotropic g = 2.02 EPR signal was shown to originate from a 3Fe cluster which may have oxygenous or nitrogenous ligands. In addition, the Mossbauer data also revealed two [4Fe-4S]2+ clusters iun each molecule of hydrogenase II. The EPR and Mossbauer data of hydrogenase I were found to be identical to those of hydrogenase II, indicating that both enzymes have common metallic centers. PMID- 6294074 TI - Isolation and identification of 25-hydroxyvitamin D3-26,23-peroxylactone. A novel in vivo metabolite of vitamin D3. AB - A new vitamin D3 metabolite was isolated in pure form (18.2 micrograms) from the serum of rats given large doses (two doses of 26 mumol/rat) of vitamin D3. The new metabolite has been unequivocally identified as 3 beta, 25-dihydroxy-9,10 seco-5,7,10(19)-cholestatrieno-26,23-peroxylactone by ultraviolet absorption spectrophotometry, Fourier transform infrared spectrophotometry, mass spectrometry, field desorption mass spectrometry, and specific chemical reaction with triphenyl phosphine. The stereochemical configuration at the C-23 and c-25 positions of the 25-hydroxyvitamin D3-26-23-peroxylactone was definitely determined to be the 23(S)25(R),25-hydroxyvitamin D3-26,23-peroxylactone is suggested for this metabolite. The isolation involved chloroform-methanol extraction and four column chromatographic procedures. The metabolite purification and elution position on these columns were followed by UV measurement at 264 nm. This metabolite was ultimately resolved from the previously known 25-hydroxyvitamin D3-26,23-lactone by high pressure liquid chromatography using a Zorbax Sil column. The 25-hydroxyvitamin D3-26,23 peroxylactone was converted upon storage at room temperature or -20 degrees C into the 25-hydroxyvitamin D3-26,23-lactone. Since under the conditions of this isolation only the 26,23-peroxylactone and no 26,23-lactone of 25-hydroxyvitamin D3 was present in the rat serum, this suggests that the 25-hydroxyvitamin D3 26,23-peroxylactone is the naturally occurring metabolite. PMID- 6294075 TI - Effect of differentiation on the adenylate cyclase system of 3T3-C2 and 3T3-L1 cells. Determination of choleragen substrates in differentiating 3T3-L1 and nondifferentiating 3T3-C2 cells. AB - 3T3-L1 preadipocytes, when treated with 3-isobutyl-1-methylxanthine, dexamethasone, and insulin, differentiate into cells with the morphological and biochemical properties of adipocytes; the closely related 3T3-C2 cells, under identical conditions, exhibit a low frequency of adipocyte conversion. During differentiation, 3T3-L1 preadipocytes acquire an increased responsiveness to certain agonists (e.g. isoproterenol and adrenocorticotropic hormone) that influence lipolysis and lipogenesis through activation of adenylate cyclase, whereas 3T3-C2 cells do not. It has been suggested that changes in hormone responsiveness of 3T3-L1 cells during differentiation result from increased amounts of the guanyl nucleotide-binding protein of adenylate cyclase, as demonstrated by choleragen-catalyzed [32P]ADP ribosylation of 42 and 49-50 kilodalton particulate peptides. Particulate fractions from nondifferentiating 3T3-C2 cells, like those from 3T3-L1 cells, contained choleragen substrates of 42 and 46-47 (doublet) kilodaltons. Incubation of intact 3T3-L1 or 3T3-C2 cells with choleragen prior to preparation of particulate fractions prevented the subsequent in vitro choleragen-dependent [32P]ADP ribosylation of only these peptides. Increased incorporation of radioactivity into both the 42 and 46-47-kilodalton peptides was observed during differentiation of 3T3-L1 cells. However, a similar increase was also observed in nondifferentiating 3T3-C2 cells subjected to the differentiation protocol. Therefore, increased hormone responsiveness of 3T3-L1 adipocytes cannot be explained solely on the basis of increased labeling, and perhaps increased amounts, of the guanyl nucleotide-binding protein. PMID- 6294076 TI - Energy transduction by the reconstituted b-c1 complex from yeast mitochondria. Inhibitory effects of dicyclohexylcarbodiimide. AB - A purified cytochrome b-c1 complex isolated from yeast mitochondria has been reconstituted into proteoliposomes. The reconstituted comp]lex catalyzed antimycin A-sensitive electron transfer from different analogues of coenzyme Q to cytochrome c. The reconstituted complex was also capable of energy conservation as indicated by uncoupler-stimulated rates of electron transfer, electrogenic proton ejection, and reversed electron flow from cytochrome b to coenzyme Q2 in the presence of antimycin A driven by a valinomycin-induced K+-diffusion potential (negative inside). Close to four protons were ejected per two electrons transported through the reconstituted b-c1 complex with ferricyanide as an artificial and impermeable electron acceptor.l The H+/2e- ratio decreased to two in the presence of the proton-conducting agent, carbonyl cyanide m chlorophenylhydrazone. The same processes were studied in parallel in energy conserving site 2 of rat liver mitochondria with similar results. In the reconstituted b-c1 complex, dicyclohexylcarbodiimide (DCCD) blocked the function of the electrogenic proton translocating device in the forward direction of proton ejection as well as in the backwards direction, measured as reversed electron flow from cytochrome b to coenzyme Q2 driven by a K+-diffusion potential. The primary effect of DCCD is localized on the proton ejection process, as the low proton conductance of the proteoliposome membrane was totally preserved after DCCD treatment. PMID- 6294077 TI - Characterization of the homopolymer tailing reaction catalyzed by terminal deoxynucleotidyl transferase. Implications for the cloning of cDNA. PMID- 6294078 TI - Studies on the interaction of palmitoyl coenzyme A with the adenine nucleotide translocase. PMID- 6294079 TI - Fibronectin binding to Staphylococcus aureus. AB - Bacteria are able to interact with a number of macromolecules which act as opsonins or tissue-adherence factors. Because soluble fibronectin may be important factors. Because soluble fibronectin may be important in the phagocytic removal of bacteria and insoluble fibronectin may serve as a bridge between bacteria and host tissues, we have characterized the binding of soluble plasma fibronectin to Staphylococcus aureus as a first step to understanding these interactions. The binding of 125I-fibronectin to clinical and laboratory strains of S. aureus was studied. Bound fibronectin was separated from free fibronectin by centrifugation. Specific binding was determined by subtracting the amount bound in the presence of excess fibronectin from the total amount bound. We found that (i) fibronectin bound saturably, irreversibly, and noncovalently to S. aureus when the binding reaction was carried out at pH 7.4 or greater; (ii) S. aureus harvested in logarithmic phase of growth from media buffered to pH 8.4, and from brain-heart infusion media which demonstrated the greatest number of fibronectin-binding sites; (iii) high molecular weight dextrans, fibrinogen, cyanogen bromide fragment 7 of collagen, cationic proteins, dibromide fragment 7 of collagen, cationic proteins, dithiothreitol, and protein A did not alt er fibronectin binding to S. aureus; (iv) nonsaturable binding occurred below pH 7.0 with peak binding occurring at pH 5.8; and (v) there were marked differences in the amounts of fibronectin that bind to different strains of S. aureus. S. aureus ATCC 25923, when harvested in logarithmic phase of growth from tryptic soy broth and tested for fibronectin binding in (2-hydroxyethyl)-1-piperazineethanesulfonic acid-buffered saline, pH 7.4, had 7500 binding sites/organism with an apparent association constant of 5.6 X 10(9) M-1. PMID- 6294081 TI - Interaction of a DNA-binding protein, the gene product of D5 of bacteriophage T5, with double-stranded DNA. Analysis by metrizamide gradient centrifugation. AB - Interactions of DNA and the gene product D5 (gpD5) of bacteriophage T5, a DNA binding protein that binds preferentially and cooperatively to double-stranded DNA, were analyzed by metrizamide gradient centrifugation. Conditions were set so that DNA and DNA protein complex sedimented to apparent equilibrium positions. DNA has a buoyant density of 1.12 g/cm3, and DNA saturated with gpD5 has a buoyant density of 1.17 g/cm3. These values are independent of DNA size and base composition in the range studied. At gpD5 concentration below the saturation value in a low ionic strength buffer, DNA distribution is bimodal, indicating cooperative binding of gpD5 to DNA. However, in the presence of 10 mM MgCl2, the binding process becomes distributive, with the buoyant density increasing linearly with the amount of gpD5 added until the saturation. From these data, one molecule of gpD5 is calculated to cover 40 base pairs at saturation. The technique as described has general applicability to the study of any interaction between DNA and dNA-binding proteins that bind in sufficient amount to cause detectable changes in buoyant density. PMID- 6294080 TI - Inhibition of HeLa cell protein synthesis following poliovirus infection correlates with the proteolysis of a 220,000-dalton polypeptide associated with eucaryotic initiation factor 3 and a cap binding protein complex. AB - Following poliovirus infection of HeLa cells, the synthesis of cellular proteins is inhibited but translation of poliovirus mRNA proceeds. The defect in the recognition of host cell mRNA may be due to a change in a cap recognition complex which, when added to an infected cell lysate, restores the ability to translate capped mRNAs. We employed immunoblotting techniques to examine initiation factors in crude lysates from uninfected and poliovirus-infected HeLa cells. Using an antiserum against eucaryotic initiation factor 3, we detected an antigen of approximate molecular weight 220,000 in uninfected cell lysates but not in infected cell lysates. Antigenically related polypeptides of 100,000 to 130,000 daltons, presumably degradation products, were detected in the infected cell lysate. The time course for degradation of the 220,000-dalton polypeptide correlates with that for inhibition of cellular protein synthesis in vivo. A portion of the population of 220,000-dalton polypeptides apparently associates with initiation factor eIF3 but is readily dissociated in buffers containing high salt. Affinity-purified antibodies against the polypeptide recognize a protein of the same size in a purified preparation of a cap binding protein complex obtained by cap-affinity chromatography. We postulate that the 220,000-dalton polypeptide is an essential component of the cap recognition complex and that its degradation in poliovirus-infected cells results in the inhibition of host cell translation. These results are in the first demonstration of a specific structural defect in an initiation factor resulting from poliovirus infection. PMID- 6294083 TI - Characterization of highly phosphorylated subcomponents of rat thymus H1 histone. AB - The phosphorylation of electrophoretically homogeneous preparations of the five major subcomponents of that thymus H1 histone by growth-associated histone kinase isolated from Ehrlich ascites tumor or Novikoff hepatoma cell chromatin results in the introduction of three to six phosphates/molecule into different subcomponents. Fully phosphorylated preparations of subcomponents 1 through 4 consist of H1 molecules containing a uniform number of phosphate groups, and run as single bands in long acid-urea gels. Fully phosphorylated preparations of subcomponent 5 consist of a mixture of molecules containing five and six phosphate groups. Phosphorylation of subcomponents 2, 4, and 5 occurs in both the NH2- and carboxyl-terminal regions of the molecules. Phosphorylation of subcomponents 1 and 3 occurs only in the carboxyl-terminal region. The central globular region of the histones is not phosphorylated. The major sites of phosphorylation in rat H1 histone subcomponents are similar to, but not entirely identical with, the major sites of phosphorylation previously characterized in total calf thymus H1, as determined by comparison of phosphopeptide maps. Highly phosphorylated rat H1 molecules, similar in phosphate content to those found in mitotic cells, have distinct chromatographic properties, compared to lightly phosphorylated molecules of the type found in interphase cells. This change in chromatographic properties appears to depend on the number of phosphate groups present in the histone rather than on the presence of phosphate in any specific sites. PMID- 6294082 TI - Spatial relationship between cytochrome a and a3. AB - We have studied the spatial relationship between cytochromes a and a3 by the enhancement of the spin relaxation of cytochrome a3-NO EPR signals by the paramagnetic a heme at 15 K. An Fe-Fe distance of 12-19A is estimated from the absence of dipolar broadening and from the observation of spin relaxation enhancement in the a3-NO complex. When this result is combined with resonance x ray diffraction data reported by Blasie et al. (Blasie, J. K., Pachence, J. M., Tavormina, A., Dutton, P. L., Stamatoff, J., Eisenberger, P., and Brown, G. (1982) Biochim. Biophys. Acta 679, 188-197) and the contribution from the exchange interaction is considered, we can limit the iron-iron distance to 12-16 A and estimate the angle between the Fe-Fe vector and mitochondrial membrane normal as 30-60 degrees. We also consider the possible effects of CuA on cytochrome a3-NO. PMID- 6294084 TI - Effects of streptozotocin diabetes, glucose, and insulin on the metabolism of type IV collagen and proteoglycan in murine basement membrane-forming EHS tumor tissue. AB - The influence of streptozotocin diabetes, glucose, and insulin on the metabolism of basement membrane collagen and proteoglycan was studied in the murine EHS tumor. No differences were found in the amino acid composition of tumors grown in diabetic and control mice, suggesting that the relative amount of collagen was not increased. Similarly, no significant differences were observed in any of the intracellular enzyme activities of collagen biosynthesis in the tumor tissue, but in diabetic mice, a significant increase was found in all four enzyme activities studied in the kidneys. High glucose concentrations increased total protein synthesis of the tumor tissue in vitro whereas insulin had no effect. Basement membrane collagen synthesis was also increased but to the same extent as protein synthesis in general. No qualitative changes were found in the pro-alpha 1(IV) and pro-alpha 2(IV) chains synthesized in vitro with or without insulin at different glucose concentrations. Hyperglycemia secondary to insulin deficiency may thus be responsible for the basement membrane thickening in diabetes but this cannot explain the relative increase in collagen content of these matrices. The incorporation of 35SO4 into proteoglycan in vitro was likewise increased at high glucose concentrations, indicating that the synthesis of this component may also be altered in diabetes. PMID- 6294085 TI - Cholinephosphotransferase in rat lung. In vitro formation of dipalmitoylphosphatidylcholine and general lack of selectivity using endogenously generated diacylglycerol. AB - Diacylglycerol was generated in vitro in rat lung microsomes by forming phosphatidic acid via sn-glycerol-3-phosphate acyltransferase followed by the hydrolysis of the phosphatidic acid by phosphatidate phosphohydrolase. Diacylglycerol concentrations of 35 to 50 nmol/mg of microsomal protein were obtained. Cholinephosphotransferase activity was determined in microsomes by measuring the conversion of endogenously generated [14C]diacylglycerol to phosphatidylcholine. Reaction rates of 14 to 16 nmol/min/mg of protein were obtained with a 30-s reaction. Diacylglycerol which was primarily dipalmitoylglycerol was produced when palmitic acid was used in the sn-glycerol-3 phosphate acyltransferase reactions. Dipalmitoylphosphatidylcholine was formed via cholinephosphotransferase from the dipalmitoylglycerol with an apparent maximal velocity of 20 nmol/min/mg of protein. When oleic acid was used instead of palmitic acid, the apparent maximal velocity for cholinephosphotransferase was 26 nmol/min/mg of protein. The apparent Km values for the two different diacylglycerol substrates were the same (28.5 nmol/mg of protein). Diacylglycerols, with different molecular species composition, were generated using a variety of fatty acids and fatty acid mixtures. The phosphatidylcholine formed from these diacylglycerols had the same molecular species profiles as the diacylglycerol used as the substrate. The relative reaction rates with the different diacylglycerols were essentially the same except when 20:4 and 22:6 fatty acids were used individually, in which case the rates were lower. We conclude that cholinephosphotransferase readily forms dipalmitoylphosphatidylcholine from endogenously generated dipalmitoylglycerol and that the cholinephosphotransferase reaction is generally nonselective for the diacylglycerol substrate. PMID- 6294086 TI - Selective hormonal activation of cyclic AMP-dependent protein kinase isoenzymes in normal and malignant osteoblasts. AB - The pattern of cyclic AMP-dependent protein kinase isoenzyme response to acute hormonal activation has been studied in cultured cells derived from rat osteogenic sarcoma and osteoblast-rich cells grown from newborn rat calvaria. Using multiple small anion exchange columns and a batch elution technique, a rapid method of separating the isoenzymes of cyclic AMP-dependent protein kinase was developed and the acute activation by parathyroid hormone and prostaglandin E2 of each isoenzyme was studied. Activation was rapid, being detectable at 5 s, maximal at 15-30 s, and persisting for up to 6 h. Both hormones showed a dose dependent activation of each isoenzyme in both cell types, but the patterns of response differed. Parathyroid hormone predominantly stimulated isoenzyme I in the clonal osteogenic sarcoma cells but showed equivalent activation of each isoenzyme in calvarial cells. Prostaglandin E2 also predominantly stimulated isoenzyme I in the malignant cells, whereas in the calvarial strain there was a major effect on isoenzyme II with almost no stimulation of isoenzyme I. Half maximal stimulation of cyclic AMP-dependent protein kinase in the malignant cell strain was achieved for both hormones at concentrations an order of magnitude lower than those in the normal strain. These studies demonstrate selective activation of cyclic AMP-dependent protein kinase isoenzymes by hormones. Furthermore, the nature of the response differs between the normal and the corresponding neoplastic cell types for the same hormone stimulus. PMID- 6294087 TI - Effects of chronic insulin and glucagon exposure on the biosynthesis of glycerolipids by cultured hepatocytes. AB - Primary cultures of adult rat hepatocytes incubated (24-72 h) in Waymouth's 752/1 medium, exhibited a marked decline (80-90%) in their ability to incorporate 0.5 mM [1,3-14C]glycerol and 1.0 mM palmitate into glycerolipids. The specific activities of glycerol kinase and sn-glycerol-3-P acyltransferase also showed a time-dependent reduction (30-80%). Phosphatidate phosphohydrolase, diacylglycerol cholinephosphotransferase, and fatty acid-CoA ligase activities were unaffected. Insulin and/or glucagon (10(-8) to 10(-6) M) not only prevented these reductions in hepatocyte monolayer glycerolipid formation and enzyme activities but increased (2-5-fold) the level of these processes. Cycloheximide (1 microM) reduced the insulin- and glucagon-dependent increases in sn-glycerol-3-P acyltransferase and glycerol kinase activity and glycerolipid biosynthesis. There was an excellent correlation (r greater than 0.93) between changes in glycerol kinase and sn-glycerol-3-P acyltransferase activity and the capacity of hepatocyte monolayers to incorporate labeled glycerol into glycerolipids under all conditions studied. Therefore, insulin and glucagon may regulate hepatic glycerolipid biosynthesis in part by maintaining the liver cell's enzymatic rate of glycerolipid biosynthesis. PMID- 6294088 TI - The granulocyte-angiotensin system. Angiotensin I-converting activity of cathepsin G. AB - Cathepsin G, an Mr = 26,000-29,000 cationic human neutrophil lysosomal serine protease, releases angiotensin II from angiotensinogen and was, therefore, examined for angiotensin I-converting activity. Cathepsin G-dependent angiotensin I conversion was detected by a high performance liquid chromatography assay which permitted independent quantitation of angiotensin I and angiotensin II and detection of angiotensin degradation products. 1.8-5.0 X 10(-9) M cathepsin G converted angiotensin I (3.3 X 10(-4) M) to angiotensin II without further degradation of angiotensin II. The pH optimum for cathepsin G-catalyzed angiotensin I conversion was pH 7.0-7.5, and the Km and Kcat were 2.2 X 10(-4) M and 3.4 s-1, respectively. In contrast to dipeptidyl hydrolase-converting enzyme, cathepsin G did not inactivate bradykinin, did not cleave hippuryl-His-Leu, and was not inhibited by 10(-4) M Captopril or SQ 20881. Purified human neutrophils stimulated with 2.5 X 10(-6) M-10(-10) M fMet-Leu-Phe released angiotensin converting activity with a Km of 3.3 X 10(-4) M. That the angiotensin-converting activity released from neutrophils was attributable to cathepsin G was indicated by similar susceptibility to inhibitors and adsorption by goat antibody to cathepsin G. The granulocyte-angiotensin system provides a mechanism for the local generation of angiotensin II at sites of neutrophil accumulation and may be of significance in regulation of blood flow in tissue microvasculature. PMID- 6294089 TI - The stereochemical configuration of Mn(II) . ADP at the active site of creatine kinase elucidated by electron paramagnetic resonance with Rp [alpha-17O]ADP and Sp [alpha-17O]ADP. AB - The stereochemical configuration of the Mn(II) . ADP complex at the active site of creatine kinase has been elucidated by EPR using chirally labeled [alpha 17O]ADP. Superhyperfine coupling between the 17O nucleus and Mn(II) produces a characteristic inhomogeneous broadening of the EPR signals for Mn(II) whenever the 17O is in the first coordination sphere of the metal ion. Previous experiments with ADP regio-specifically labeled with 17O in the phosphate groups had shown that Mn(II) was coordinated to the alpha and beta positions of ADP in transition state analog complexes with creatine kinase that involved enzyme, Mn(II), ADP, creatine, and anions such as formate and thiocyanate (Reed, G. H., and Leyh, T. S. (1980) Biochemistry 19, 5472-5480). The present experiments were initiated to determine the stereochemical configuration of the Mn(II) . ADP complex at the active site of the enzyme. EPR spectra for Mn(II) in transition state analog complexes with formate, thiocyanate, and nitrate as the stabilizing anions show inhomogeneous broadening from 17O in Sp [alpha-17O]ADP whereas the spectra obtained with Rp [alpha-17O]ADP are indistinguishable from those for matched samples with unlabeled ADP. The precision of the measurements indicates that the stereoselectivity of the enzyme for the delta configuration of the alpha, beta chelate of Mn(II) . ADP is greater than 15:1. The delta configuration is also preferred in fully active enzymic complexes involving the equilibrium mixture of substrates. PMID- 6294090 TI - Nucleotide sequence of the yeast nuclear gene for cytochrome c peroxidase precursor. Functional implications of the pre sequence for protein transport into mitochondria. PMID- 6294091 TI - Enzymatic deglycosylation of the subunits of chorionic gonadotropin. Effects on formation of tertiary structure and biological activity. AB - Both the O- and N-linked oligosaccharide moieties of the subunits of the placental glycoprotein hormone, human choriogonadotropin (hCG), are removed by treatment with a mixture of glycosidases produced by Streptococcus (Diplococcus) pneumoniae. The resulting deglycosylated subunits recombine with their native counterparts in good yield, and the reassociated hormones bind to gonadotropin receptors equally as well as the untreated hormone. Stimulation of steroidogenesis by the deglycosylated alpha-native beta recombinant, however, was markedly less than the stimulation by unmodified hCG both in terms of relative potency (0.10-0.15) and the maximal amount of steroid (40-50%) produced. The native alpha-deglycosylated beta recombinant produced a maximum level of steroid production of 80-90% that of control hCG although its relative potency had decreased approximately 4-fold. The data are in accord with results by others in which either hCG or lutropin was partially deglycosylated by treatment with anhydrous hydrofluoric acid. In addition, the effects of deglycosylation on the ability of each subunit to refold after reduction of their disulfide bonds was studied. Of particular interest is that, after deglycosylation, the beta subunit can correctly refold to a significant degree, in contrast to several unsuccessful attempts to demonstrate correct refolding of the unmodified beta subunit of either lutropin or hCG. Alpha subunit, as measured by a conformation sensitive radioimmunoassay, refolds with equal facility both before and after deglycosylation. PMID- 6294092 TI - The gamma subunit of 7 S nerve growth factor binds to cells via complexes formed with two cell-secreted nexins. PMID- 6294093 TI - Phosphorylation of a membrane receptor for glycoproteins. Possible transmembrane orientation of the chicken hepatic lectin. AB - The chicken hepatic lectin, a receptor for partially deglycosylated serum glycoproteins, has been identified as a phosphoprotein. Phosphorylation was detected by incorporation of 32P into the protein in cultured hepatocytes and by two-dimensional gel analysis of protein purified from liver tissue. In addition, forms of the receptor containing one, two, and three sialic acid residues have been detected, with the disialylated form predominating. The site of phosphorylation has been identified as Ser7 in the complete amino acid sequence of the receptor (Drickamer, K. (1981) J. Biol. Chem. 256, 5827-5839). The presence of a protein kinase target site near the NH2-terminal of the receptor, a stretch of 25 uncharged, hydrophobic residues in positions 24 through 48, and a site of glycosylation at position 67 suggests that the chicken hepatic lectin is probably a transmembrane protein, oriented with COOH-terminal outside the cell and NH2-terminal in the cytoplasm. PMID- 6294094 TI - Structural analysis of sericin genes. Homologies with fibroin gene in the 5' flanking nucleotide sequences. AB - Two closely related mRNAs (11.0 and 9.6 kilobases long) for sericin, a cocoon protein other than fibroin, were isolated from the middle silk gland of Bombyx mori. They were shown to be specifically accumulated in this particular part of the gland. Nine genomic clones complementary to the mRNAs were isolated from the recombinant phage library of Bombyx DNA. They represent two structurally similar genomic segments, and contain the 5' end and its surrounding region of sericin gene. We have identified at least 5 exons in then, and the intervening coding junctions were determined at nucleotide level. The two genomic segments differ mainly in the 5' flanking as well as in the intervening sequences and may represent alleles of a sericin locus. DNA sequence analysis of an exon has revealed an internally repetitive unit of 114 base pairs. From an internally repetitive unit of 114 base pairs. From an open frame of the repetitive DNA sequence, a peptide consisting of 38 amino acids abundant in serine was deduced. We propose the peptide represents a repetitive unit of a sericin protein. The probable site of initiation of sericin gene transcription in vivo has been identified at the nucleotide level that also turned out to be the 5' end of mature sericin mRNA. There exist strong homologous regions in the sericin and fibroin genes at their corresponding 5' flanking sequences. A possible involvement of the homologous sequences around -111 and -186 for the concerted expression of the sericin and fibroin genes in the silk glands is discussed. PMID- 6294095 TI - Prostatic steroid-binding protein. Isolation and characterization of C3 genes. AB - Prostatic steroid-binding protein, whose expression is stimulated by androgens, consists of two subunits, one containing the polypeptides C1 and C3 and the other containing the polypeptides C2 and C3. We have isolated and sequenced cDNA clones specific for C3 mRNA and used them to isolate and characterize genomic clones for two C3 genes. Both genes are 3.2 kilobases with identical exon/intron arrangements, which is similar to the organization of the C1 and C2 genes, suggesting that they may have arisen by duplications of an ancestral gene. Finally, homologous human genes have not been detected. PMID- 6294096 TI - The primary structure of the alcohol dehydrogenase gene from the fission yeast Schizosaccharomyces pombe. AB - We have cloned and sequenced the alcohol dehydrogenase gene of the fission yeast Schizosaccharomyces pombe. The gene was isolated by transformation and complementation of a Saccharomyces cerevisiae strain which lacked functional alcohol dehydrogenase with an S. pombe gene bank constructed in the autonomously replicating yeast plasmid YEp13. Southern hybridization analysis indicates that S. pombe contains only one alcohol dehydrogenase gene. The structural region of the gene is 50% homologous to the alcohol dehydrogenase encoding genes of the budding yeast S. cerevisiae. The gene exhibits a very strong codon usage bias; with the set of predominantly used codons generally resembling that which S. cerevisiae employs preferentially. All of the differences in codon usage bias between S. pombe and S. cerevisiae are in the direction of greater G + C content in S. pombe codons. It is argued that this observation supports the hypothesis that selection toward uniform codon-anticodon binding energies contributes to codon usage bias and that the optimum binding energy is, on the average, higher in S. pombe than S. cerevisiae. PMID- 6294097 TI - Demonstration by covalent cross-linking of a specific interaction between beta endorphin and calmodulin. PMID- 6294098 TI - Protein C inhibitor. Purification from human plasma and characterization. AB - Protein C inhibitor was isolated from human plasma using conventional chromatographic technique consisting of barium citrate adsorption, polyethylene glycol fractionation, DEAE-Sepharose CL-6B treatment, ammonium sulfate fractionation, dextran sulfate-agarose chromatography, gel filtration on ACA-44, and DEAE-Sephacel chromatography. The purified protein C inhibitor is a single polypeptide chain with an apparent Mr = 57,000 on sodium dodecyl sulfate polyacrylamide gel electrophoresis. The inhibitor is heterogeneous in pI: six pIs exist between pH 7.4 and 8.6. The inhibitor was shown to be different from the already known plasma protease inhibitors by chemical and immunological analyses. It migrates to the late alpha 1-globulin region on agarose gel electrophoresis. The inhibitor reduced the amidolytic activity of activated protein C noncompetitively by forming a 1:1 molar complex with the enzyme, determined by the use of a fluorogenic substrate toward activated protein C (Boc-Leu-Ser-Thr Arg-4-methylcoumaryl-7-amide). The inhibition constant (Ki) of the inhibitor against activated protein C was 5.8 x 10(-8) M. The inhibitor also blocked the prolongation of activated partial thromboplastin time by activated protein C. The immunoglobulin which was produced by the inhibitor completely removed the inhibitory activity present in normal human plasma against activated protein C. This suggests that the inhibitor which we have isolated is the only inhibitor in plasma against activated protein C. PMID- 6294099 TI - Conformational and biological properties of di[delta-(5-nitro-2 pyrimidyl)ornithine 17,18]glucagon. Role of the arginine residues. AB - The reaction of nitromalondialdehyde with the arginine residues of glucagon results in the conversion of the 2 arginine residues in the peptide to delta-(5 nitro-2-pyrimidyl)ornithine to form di[delta-(5-nitro-2-pyrimidyl)ornithine 17,18]glucagon (NP-glucagon). The modified peptide does not exhibit any loss in ability to activate adenylate cyclase of rat liver plasma membranes or to stimulate glycogenolysis in cortisone-primed rabbits relative to the native hormone despite this marked alteration in structure. The CD of dilute solutions of NP-glucagon is similar to that of the native hormone. In the absence of salt, the CD of NP-glucagon is independent of peptide concentration, but structures of higher helical content are observed in concentrated peptide solutions in the presence of 0.1 M NaCl and in methanol. The extent of helix formation under these conditions is greater than that given by glucagon. Results from viscosity and proton magnetic resonance spectra confirm and extend previous studies to indicate that this fully active derivative is in a compact folded conformation. PMID- 6294100 TI - Electron transfer across the chromaffin granule membrane. AB - Membrane vesicles (ghosts) containing ascorbic acid were prepared from bovine chromaffin granules. When ferricyanide or ferricytochrome c were added to the external medium, a membrane potential (interior positive) developed across the ghost membrane. This membrane potential could not be elicited from ascorbate-free ghosts or by ferrocyanide added instead of ferricyanide. These results indicate that the chromaffin-granule membrane has a transmembrane electron carrier with a midpoint potential between that of ascorbate (+85 mV) and that of cytochrome c (+255 mV). The most likely candidate is cytochrome b-561 (+140 mV). PMID- 6294101 TI - Structure and organization of a cluster of sic tRNA genes in the space between tandem ribosomal RNA gene sets in Bacillus subtilis. AB - Hybridization of Southern blots of EcoRI digests of total Bacillus subtilis DNA with ribosomal RNA and transfer RNA probes provides evidence of highly clustered tRNA and rRNA genes, with several tRNA clusters being located in spaces which span between tandem ribosomal RNA gene sets. Clones containing these tRNA clusters were isolated from a Charon 4A library. One of them, denoted trrnB, was partially sequenced. A cluster of six tRNA genes was found, with anticodon assignments of Asn, Thr, Gly, Arg, Pro, Ala. This cluster is closely flanked on both sides by ribosomal RNA gene sets, which were identified by sequencing upward through the 5 S rRNA gene, across a space, and into the 23 S rRNA gene, and also sequencing downward into the 16 S rRNA gene. The tRNA gene cluster appears to be organized into at least two transcriptional units separated by an attenuator region. These transcriptional units may be components of the flanking ribosomal RNA operons. The putative promoter region of the downstream 16 S rRNA is organized differently from Escherichia coli; it is smaller and seems less complex. This gene organization provides insight into possible mechanisms for coordinate and differential control of transfer and ribosomal RNA gene expression. PMID- 6294102 TI - Binding sites for lactogenic and somatogenic hormones from rabbit mammary gland and liver. PMID- 6294103 TI - Selective affinity labeling and molecular characterization of hepatic alpha 1 adrenergic receptors with [3H]phenoxybenzamine. PMID- 6294104 TI - Location of 5.8 S rRNA contact sites in 28 S rRNA and the effect of alpha-sarcin on the association of 5.8 S rRNA with 28 S rRNA. AB - We have constructed phage M13mp7 clones each containing the coding strand from one of three restriction fragments which collectively span the mouse 28 S rRNA gene with the exception of the 3'-terminal approximately 500 base pairs. When hybridized to 28 S rRNA, only the fragment containing the 5'-terminal 1400 nucleotides of the gene inhibited the annealing of 5.8 S rRNA to the 28 S rRNA. The same results were obtained when either the 5'- or 3'-terminal fragment of 5.8 S rRNA was used in lieu of intact 5.8 S rRNA, each of which had been shown to contain an independent 28 S rRNA contact site. However, alpha-sarcin, a cytotoxin that inhibits protein synthesis by hydrolyzing a phosphodiester bond near the 3' end of 28 S rRNA, produces a 3'-terminal 488-nucleotide fragment which exhibits a marginal capacity to anneal to 5.8 S rRNA. These results indicate that 5.8 S rRNA interacts predominantly with a structural domain near the 5' end of 28 S rRNA. This conclusion is consistent with base-pairing interactions between 5.8 S rRNA and 28 S rRNA based on the proposed secondary structures for Escherichia coli 23 S and yeast 26 S rRNAs. However, alpha-sarcin treatment of ribosomes affects the stability of the binding of 5.8 S rRNA to the 28 S rRNA, even though the toxin hydrolyzes a phosphodiester bond several thousand nucleotides from the proposed contact regions. Finally, mouse 5.8 S rRNA was shown to lack two internal nucleotides reported to be present in rat 5.8 S rRNA. PMID- 6294105 TI - Evidence of an ubisemiquinone radical(s) from the NADH-ubiquinone reductase of the mitochondrial respiratory chain. AB - NADH-ubiquinone (Q) reductase isolated from beef heart mitochondria exhibited, upon reduction by NADH, a prominent EPR signal at room temperature attributable to stable ubisemiquinone radical(s). The concentration of the ubisemiquinone radical reached as high as 40% of the total Q content in the reductase. The radical was virtually abolished by adding rotenone, whereas rotenone had no effect on the reduction of FMN by NADH. The radical showed an EPR signal of g = 2.0042 at approximately 9.5 GHz with no resolved hyperfine structure and had a line width of 6.8 Gauss at 23 degrees C. The Q-band EPR spectra at 35 GHz showed well resolved g-anisotropy and had a field separation between derivative extrema of 24 Gauss. These results substantiate the fact that this radical was bound to a protein; we call it ubiquinone protein-N (QP-N). The pH dependence of the EPR signals demonstrated that the species of the ubisemiquinone radical(s) consisted of not only an anionic form but also a neutral form. Only about half of the QP-N radical formed by NADH reduction was abolished by p-chloromercuric sulfonate. The microwave power saturation curve of the radical was biphasic; the first phase leveled off at about 5 milliwatts and then at about 20 milliwatts. These results suggested that the ubisemiquinone radical from QP-N was heterogenous, consisting of at least two populations of stable ubisemiquinone radical(s). It is suggested that two kinds of QP-N exist in NADH-Q reductase. Each mole of protein may bind two mol of Q. PMID- 6294106 TI - Tandem repeat of the genes for protein S, a development-specific protein of Myxococcus xanthus. AB - Protein S, a development-specific protein of Myxococcus xanthus is produced only during fruiting body formation. More than 15% of total protein synthesis during this period is accounted for by the production of protein S. The genes for protein S were identified and cloned with the use of mixed probes consisting of eight synthetic oligodeoxyribonucleotides (tetradecamers) which correspond to a carboxyl-terminal portion of protein S. The two genes are oriented in the same direction and are separated approximately 1.2 kilobases. The DNA sequences of the carboxyl-terminal portions of the two genes reveal that both can code for the identical eleven amino acid sequence which corresponds to the carboxyl-terminal end of protein S. However, there are a few base substitutions upstream of these regions. This duplication of genes in M. xanthus may facilitate the extremely rapid synthesis of protein S during fruiting body formation. PMID- 6294107 TI - Separation of multiple phosphotyrosyl-and phosphoseryl-protein phosphatases from chicken brain. PMID- 6294108 TI - Pyruvoyl-dependent histidine decarboxylases from Clostridium perfringens and Lactobacillus buchneri. Comparative structures and properties. AB - Histidine decarboxylase (EC 4.1.1.22) was purified to homogeneity from Clostridium perfringens and also from Lactobacillus buchneri. Both enzymes are composed of alpha and beta subunits, with an essential pyruvoyl group bound to the alpha subunit. In this respect and also in molecular weight of both the alpha and beta subunits and the native enzyme, they closely resemble the previously described (Riley, W.D., and Snell, E. E. (1970) Biochemistry 9, 1485-1491) histidine decarboxylase from Lactobacillus 30a. Rabbit antibodies to the latter enzyme cross-react incompletely with the decarboxylase from L. buchneri but not with that from C. perfringens in double diffusion tests. The clostridial decarboxylase differs substantially from the Lactobacillus 30a enzyme in amino acid composition and, unlike the latter enzyme, requires high ionic strength (I approximately 1.4 M) for maximum activity. The enzymes also differ in rates of electrophoretic migration. A proenzyme for the decarboxylase similar to that previously found in Lactobacillus 30a was detected in immunoprecipitates of extracts of L. buchneri. We conclude that these proteins arise from pyruvate-free precursor proteins by similar mechanisms and probably have diverged from a common ancestral protein. PMID- 6294109 TI - Hormonal antagonistic properties of chemically deglycosylated human choriogonadotropin. AB - The biological properties of chemically deglycosylated human choriogonadotropin (DG-hCG) preparations were examined in collagenase-dispersed rat interstitial cells in vitro. Despite effective receptor binding activity in membrane preparations, DG-hCG failed to induce cyclic AMP accumulation in the cells when incubated in the presence or absence of a phosphodiesterase inhibitor. The steroidogenic ability as assessed by testosterone accumulation in the medium was less than 0.5% of the native hormone with a failure to attain maximal steroid production. Time course experiments have revealed that altered kinetics could not be responsible for the loss of hormone response. Consistent with its property of good receptor binding and poor cell activation, DG-hCG antagonized the action of native hCG. When added to the cells at the same time, DG-hCG inhibited the action of a maximal stimulatory dose of hCG in a dose-dependent manner. Inhibition of cyclic AMP accumulation was complete whereas inhibition of steroidogenesis was about 75%. DG-hCG had no effect on the stimulatory action of cholera toxin in interstitial cells or that of follitropin in rat seminiferous tubular preparations. The data suggest that DG-hCG has a conformation conductive for effective interaction with the receptor, but its ability to activate the adenylate cyclase is either lost or weakly expressed. PMID- 6294110 TI - Evidence for cAMP-independent inhibition of S-phase DNA synthesis by prostaglandins. AB - Two prostaglandins, prostaglandin E1 (PGE1) and prostaglandin B1 (PGB1), block S phase DNA synthesis in synchronous cultured baby hamster kidney (BHK) cells. The prostaglandin inhibition of DNA synthesis does not appear to require elevated levels of cAMP. In BHK-21 cells that have been "desensitized" to prostaglandin stimulation of adenylate cyclase and, therefore, have control levels of cAMP, PGE1 retains its inhibitory effect on the incorporation of tritiated thymidine into DNA. When BHK cells are exposed to PGB1 (a prostaglandin that does not elicit a cAMP response), DNA synthesis is also blocked. In nonsynchronous cells exposed for 1 h to PGE and then incubated for 1 h with PGE removed, a rebound of DNA synthesis occurs, therefore providing evidence that a transient rise of cAMP in itself is not capable of causing a cascade of reactions that block the synthesis of DNA. In addition, the concentration of PGE required for inhibition of DNA synthesis is significantly less than that required for cAMP generation. Addition of 1 x 10(-8) M PGE to BHK cells can be shown to significantly inhibit DNA synthesis within 30 min, with half-maximal inhibition seen at 3 x 10(-7) M PGE. Cyclic AMP levels for controls were 4.9 +/- 0.2 and 4.6 +/- 0.1 for 1 x 10( 6) M PGE1. These findings suggest that the prostaglandins can act independently of cAMP at physiological concentrations; and, therefore, it is possible that prostaglandins have a physiological role in the control of cell growth during S phase. PMID- 6294111 TI - Studies on the phi X174 gene A protein-mediated termination of leading strand DNA synthesis. AB - Recombinant RF (replicate form) I DNAs containing the bacteriophage phi X174 gene A protein-recognition sequence are cleaved by the phi X A protein yielding a phi X RF II X A protein complex (Zipursky, S.L., Reinberg, D., and Hurwitz, J. (1980) Proc. Natl. Acad. Sci. U.S.A. 77, 5182-5186). Such complexes support DNA synthesis in both RF I leads to SS(c) and RF I leads to RF I phi X DNA replication reactions in vitro. Two phi X A protein-recognition sequences were inserted into plasmid pBR322. Both sequences were contiguous with the same strand of the vector DNA and separated by 667 and 4275 base pairs. This recombinant plasmid (G27-4) was cleaved by the phi X A protein at either insert and both inserts support the initiation of RF leads to SS(c) DNA synthesis. This was verified by the finding that replication products were circular molecules of 667 and 4275 nucleotides. This finding is in keeping with the multifunctional activities associated with the phi X A protein; these include the site-specific nicking of RF I DNA which initiates DNA synthesis and site-specific termination resulting in the circularization of the displaced DNA strand. The phi X A protein and the Escherichia coli rep and SSb proteins catalyze the unwinding of phi X RF I DNA in vitro (Scott, J.F., Eisenberg, S., Bertsch, L.L., and Kornberg, A. (1977) Proc. Natl. Acad. Sci. U.S.A. 74, 193-197). Recombinant plasmid G27-4 RF I DNA was also unwound in vitro by this enzyme system; in this case, both circular and linear single-stranded DNA molecules of 667 and 4275 nucleotides, as well as full length circular single-stranded DNA were formed. Full length linear DNA was not detected. The two single-stranded circular DNA products formed as leading strands in RF leads to SS(c) reaction mixtures containing G27-4 RF I DNA differed in their ability to support lagging strand DNA synthesis. It was shown that the large single-stranded circular product included DNA sequences homologous to a replication factor Y effector sequence required for RF leads to RF and SS(c) leads to RF replication (Zipursky, S.L., and Marians, K.J. (1980) Proc. Natl. Acad. Sci. U.S.A. 77, 6521-6525). The 4275-nucleotide, but not the 667 nucleotide, single-stranded circular DNA product was converted to a duplex structure. PMID- 6294112 TI - Protein phosphorylation in chick kidney. Response to parathyroid hormone, cyclic AMP, calcium, and phosphatidylserine. AB - The regulation of endogenous protein phosphorylation by parathyroid hormone (PTH) was investigated using confluent monolayer cultures of chick kidney cells. Homogenates and subcellular fractions of PTH (bovine 1-34)-treated cells were subjected to an endogenous protein phosphorylation assay using ((gamma- 32P]ATP in the presence or absence of 2.0 microM cAMP or 0.5 mM Ca2+ with 25 micrograms/ml of phosphatidylserine and reactions terminated with sodium dodecyl sulfate. In other experiments, cultures were incubated in a phosphate-free 4-(2 hydroxyethyl)-1-piperazineethanesulfonic acid-buffered saline containing 50 muCi/ml of [32P]PO4 and incubations were terminated with sodium dodecyl sulfate. Protein phosphorylation was assessed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography. Cyclic AMP stimulated 32P incorporation into proteins having molecular weights of 17,000, 22,000, 35,000, 42,000, 54,000, 75,000, 80,000, 120,000, and 143,000. Calcium-phosphatidylserine stimulated the phosphorylation of proteins of 20,000, 52,000, 58,000, 60,000, and 143,000. The protein phosphorylation patterns in cultured kidney cells and freshly isolated kidney tissue were quite similar. Treatment of cultured cells with 5-50 ng/ml of PTH resulted in stimulated phosphorylation of the 35,000 and 42,000 dalton proteins as assessed by endogenous phosphorylation in homogenates. In intact cells incubated with [32P]PO4, PTH stimulated most noticeably the phosphorylation of the 35,000-dalton protein. Based on studies with cultured and fresh kidney cells, the majority of the substrate proteins for cAMP and calcium-dependent protein kinases were located in the cytoplasm with the exception of the 42,000 dalton protein which was located in the brush-border-plasma membrane fraction. The cytoplasmic cAMP-dependent protein kinase activity was responsible for the majority of PTH-stimulated protein phosphorylation. PMID- 6294113 TI - Diadenosine 5', 5"'-P1,P4-tetraphosphate stimulates processing of adp-ribosylated poly(ADP-ribose) polymerase. AB - The effect of diadenosine 5', 5"'-P1,P4-tetraphosphate (Ap4A) on the time course and acceptors of poly(ADP-ribose) synthesis was studied in undamaged and N-methyl N'-nitro-N-nitrosoguanidine-treated human lymphocytes. Analysis of protein acceptors of poly(ADP-ribose) revealed that treatment with Ap4A stimulated ADP ribosylation of bands at molecular weights of 96,000, 79,000, and 62,000. Pulse chase studies showed that these bands were produced as a result of an effect of Ap4A on the processing of ADP-ribosylated proteins rather than on the synthesis of newly ADP-ribosylated proteins. By incubating permeabilized cells in the absence or presence of Ap4A and purified poly(ADP-ribose) polymerase auto-ADP ribosylated with [32P]NAD+, we showed that the Mr = 96,000, 79,000, and 62,000 bands were derivatives of the prelabeled enzyme. Our results indicate that normal human lymphocytes process auto-ADP-ribosylated poly(ADP-ribose) polymerase to specific lower molecular weight products and that this processing is stimulated by Ap4A. PMID- 6294114 TI - Isolation of type IV procollagen-like polypeptides from glomerular basement membrane. Characterization of pro-alpha 1(IV). AB - Type IV procollagen-like constituents of glomerular basement membrane were solubilized by reduction and alkylation of disulfide bonds under denaturing conditions. Four polypeptides were observed with apparent Mr = 185,000, 175,000, 164,000, and 152,000. The two largest chains correspond to pro-alpha 1(IV) and pro-alpha 2(IV), described in model systems which secrete a basement membrane like matrix, while the smaller chains appear to be shortened forms of these polypeptides. Fractionation of the four polypeptides into two groups was achieved by ion exchange chromatography. Pro-alpha 1(IV) and 164,000 polypeptide are relatively acidic with respect to pro-alpha 2(IV) and 152,000 polypeptide, which is due in part to a relatively high content of arginine in the latter. Based on amino acid analysis of the collagenase-sensitive regions of these polypeptides, pro-alpha 1(IV) is the parent molecule from which alpha 1(IV) is derived on pepsin digestion of basement membranes and pro-alpha 2(IV) is the parent molecule of alpha 2(IV). Pro-alpha 1(IV) was isolated by gel filtration and ion exchange chromatography and characterized. It has a molecular weight of 194,000 as determined by sedimentation equilibrium. The polypeptide contains 14% carbohydrate in the form of both disaccharide, glucosylgalactosylhydroxylysine, and heteropolysaccharide units. The polypeptide backbone mass is calculated to be 167,000 daltons. Digestion of pro-alpha 1(IV) with bacterial collagenase resulted in two resistant segments of mass = 31,000 and 33,000 dalton, which make up approximately 30% of the polypeptide. PMID- 6294115 TI - Transcriptional regulation of two genes specifically induced by glucose starvation in a hamster mutant fibroblast cell line. AB - This report concerns the characterization of the RNA transcripts encoded by two cDNA sequences p4A3 and p3C5, derived from a hamster temperature-sensitive mutant cell line K12. Using the two cDNA sequences as hybridization probes, we show that they occur as single copy genes in the hamster genome and encode for RNA transcripts which are highly inducible in K12 cells at 40.5 degrees C. After incubation at 40.5 degrees C for 16 h, there is a 10-fold increase in the p4A3 and p3C5 mRNA levels, reaching a final concentration of about 1% of the cytoplasmic polyadenylated RNA. We demonstrate that the kinetics of transcription of p4A3 and p3C5 directly parallel the accumulation of the mRNA levels at 40.5 degrees C. Thus, our data indicate that the expression of these two genes are primarily regulated at the transcriptional level. In addition, there is a 3- to 4 fold increase in the p4A3 and p3C5 mRNA levels when the cells are specifically starved of glucose. This implies that the expression of these two genes are stringently regulated by the availability of glucose in the culture medium. The relationship between the cDNA clones and two glucose-regulated proteins which are overproduced in K12 cells at 40.5 degrees C is discussed. PMID- 6294116 TI - Assembly of the mitochondrial membrane system. Characterization of the oxi2 transcript and localization of its promoter in Saccharomyces cerevisiae D273-10B. AB - The oxi2 gene of yeast mitochondrial DNA was previously shown to code for subunit 3 of cytochrome oxidase (Thalenfeld, B.E., and Tzagoloff, A. (1980) J. Biol. Chem. 255, 6173-6180). In Saccharomyces cerevisiae D273-10B, a 3.6-kilobase (kb) transcript has been mapped to the oxi2 region of mitochondrial DNA. This transcript, presumed to be the messenger RNA of subunit 3, has been characterized by Northern hybridization analysis and by S1 nuclease mapping. The 3.6-kb transcript has a 5' untranslated leader of 490 nucleotides followed by a 807 nucleotide long coding sequence and a 3' extension of approximately 2450 nucleotides. The nucleotide sequence of the coding region in the 3.6-kb transcript is identical with the gene sequence, thus excluding the presence of introns in the oxi2 gene. Analysis of mitochondrial RNA in cytoplasmic petite mutants containing the oxi2 gene, but with varying lengths of flanking sequences, suggest the presence of a common promoter for oxi2 and the upstream valine tRNA. The promoter has been mapped to a 400-nucleotide long region located on the 5' side of the tRNA gene. Generation of the mature subunit 3 mRNA must, therefore, involve the excision of the tRNA from the primary transcript. PMID- 6294117 TI - Photoaffinity labeling of the N-formyl peptide receptor binding site of intact human polymorphonuclear leukocytes. A label suitable for following the fate of the receptor-ligand complex. PMID- 6294118 TI - Role of carbohydrate in human chorionic gonadotropin. Effect of deglycosylation on the subunit interaction and on its in vitro and in vivo biological properties. PMID- 6294119 TI - Characterization of sarcomeric myosin heavy chain genes. AB - Myosin heavy chain is encoded by a large multigene family. Using pMHC-25, a recombinant cDNA clone isolated from the rat myogenic cell line L6E9, four members of this family in the rat have been isolated and shown to be tissue specific and developmentally regulated. The coding regions of these genes share regions of homology interspaced with regions of non-homology. Detailed analysis of one embryonic and one adult myosin heavy chain gene shows that the coding sequences are interrupted by numerous intervening sequences whose number, size, and distribution do not appear to be conserved in the same organism or between species. PMID- 6294120 TI - An in-vitro study of the effect of buffer on the degradation of poly(glycolic acid) sutures. AB - The tensile strengths of poly(glycolic acid) (PGA) sutures immersed in buffered and unbuffered aqueous media were compared. The media used were an unbuffered physiological saline solution (pH = 5.0) and a phosphate-buffered physiological saline solution (pH = 7.4). PGA samples were immersed for various periods in each medium, and kept at 37 +/- 1 degree C in a constant temperature oven. The tensile strengths of the specimens were tested immediately after removal from the medium. Stress-strain curves of the specimens were expressed in terms of the stress unit "tenacity," commonly used in the study of fibrous polymers; it is an appropriate unit for materials of fibrous nature. These stress-strain curves were investigated as functions of buffering and duration of immersion. Degradation reduced the tensile strength of PGA more in the buffered saline solution than in the unbuffered. This higher rate of degradation in the buffered solution might be due to the presence of Na2HPO4, which removed the degradation products, shifted the reaction toward increased hydrolosis, and accelerated the loss of tensile strength in the PGA. A continuous decrease in the pH of the unbuffered solution supports this explanation. Tied-chain segments of macromolecules, a theory widely used in the study of mechanical strength of fibrous polymer may be the key to a comprehensive description of the degradation phenomenon of PGA. PMID- 6294121 TI - The characteristics of an anchorage-independent clonal agar assay for primary explanted bovine granulosa cells. AB - Normal, primary explanted, bovine granulosa cells grow reproducibly in agar culture as anchorage-independent clones. Epidermal growth factor (EGF) and rat erythrocytes are effective stimulators of colony formation, and when both are added to the culture medium at optimal concentrations, there is an enhancement of colony numbers and colony size, indicative of an independent, and operationally additive, mode of action for the two factors. The ability of cells propagated from agar clones to secrete progesterone, and to augment progesterone secretion 4 fold in the presence of 1 mM dbcAMP is proof that colonies originate from and are composed of functional granulosa cells. Maximal colony numbers are present at day 10 of incubation, and colony forming cells undergo self-renewal as assessed by the ability of cells from primary colonies to reclone in agar. Absolute cloning efficiency, however, is dependent on a number of factors. Inherent variability exists in cloning efficiency of granulosa cells from individual follicles. Quantitative and qualitative clonal growth was improved at an osmolality of less than 300 mOsm when compared with higher osmolalities. Cl-1 medium and the alpha modification of Eagle's medium were equally effective in supporting agar clonogenic growth, whereas both Ham's F12 and NCTC 135 media exhibited poor clonogenic growth supporting properties. The substitution of agarose for agar did not affect colony numbers but colonies grown in the presence of agarose tended to be smaller and more uniform in size. PMID- 6294122 TI - Amiloride, protein synthesis, and activation of quiescent cells. AB - Amiloride is known to inhibit both influx of sodium ions and activation of quiescent cells by growth factors. The coincidence of these effects has been cited to support the proposal that influx of sodium ions acts as a mitogenic signal. Although it was noted that amiloride inhibited protein synthesis, this was attributed to an action on transport of amino acids, particularly those coupled to sodium fluxes. We find, however, that amiloride directly inhibits polypeptide synthesis in a reticulocyte lysate. In Swiss 3T3 cells, concentrations of amiloride and of cycloheximide that are nearly matched in their degree of inhibition of protein synthesis, produce about the same degree of inhibition of transit of cells from G0 to S. Inhibition of protein synthesis is sufficient to explain the effect of amiloride on mitogenesis; the drug, therefore, is not suitable for testing the hypothesis that sodium influx is a mitogenic signal. PMID- 6294123 TI - Selective utilization of omega 6 and omega 3 polyunsaturated fatty acids by human skin fibroblasts. AB - Incorporation of exogenous [14C] arachidonate by human skin fibroblasts was found to be significantly greater than that of either [14C]linoleate or alpha-[14C] linolenate. Arachidonate was preferentially esterified in the PI + PS and PE classes of phospholipids. Over 40% of the incorporated [14C] arachidonate was chain elongated in 24 hours. Cells were also grown in lipid-free medium to enhance PUFA desaturation and elongation and the utilization of various omega 6 and omega 3 metabolites examined. Whereas [14C] linoleate partitioned approximately 50:50 between PL and TAG, eicosatrienoate (20:3 omega 6) was selectively sequestered in TAG. Arachidonate and docosatetraenoate (22:4 omega 6) were preferentially incorporated into phospholipids; the PI + PS fraction was most highly enriched with arachidonate. Modification of alpha-[14C] linolenate was more extensive than that of [14C] linoleate. Docosapentaenoate (22:5 omega 3) was the major omega 3 [14C] PUFA of PI + PS and PE. Eicosapentaeonate was not selectively incorporated into phospholipids; within phospholipids the 20:5 omega 3 was primarily in PC. These results indicate that human skin fibroblasts exhibit acyl specificity in the esterification of polyunsaturated fatty acids, including preferential utilization of arachidonate rather than other prostaglandin precursors in the PI + PS fraction. PMID- 6294124 TI - Cells transformed by Rous sarcoma virus release transforming growth factors. AB - Chicken embryo fibroblasts and hamster BHK cells transformed by Rous sarcoma virus (RSV) release in their culture media growth factors which enhance markedly anchorage-independent colony formation in gelified medium, at the restrictive temperature (41 degrees 5 C), of chicken embryo fibroblasts (CEF) infected by RSV mutants with a ts mutation of the src gene. This action is not observed with uninfected CEF, and, therefore, appears to require some expression of the viral src gene in the target cells. The enhancing factors are proteins related to the family of the transforming growth factors (TGFs) by their molecular weight (about 20 kd), their heat and acid resistance, and their sensitivity to dithiothreitol. They do not compete with 125I EGF for binding on the EGF receptors of the membrane of A431 cells. As chicken embryo fibroblasts are devoid of EGF receptors, their activity is not potentiated by EGF. PMID- 6294125 TI - Factors involved in supporting the growth and steroidogenic functions of bovine adrenal cortical cells maintained on extracellular matrix and exposed to a serum free medium. AB - Bovine adrenal cortex cells maintained on extracellular matrix (ECM)-coated dishes will proliferate actively when serum is replaced by HDL (25 micrograms protein/ml), insulin (10 ng/ml), and FGF (100 ng/ml). The cells have an absolute requirement for HDL in order to survive and grow. The omission of insulin, FGF, or both results in a slower growth rate and lower final cell density of the cultures. A requirement for transferrin (1 microgram/ml) becomes apparent only when cells have been grown for at least four generations in the absence of serum. Early passage (P1-P3) bovine adrenal cortex cells cultured in serum-free medium responded to ACTH (10(-8)M) with increased 11-deoxycortisol production; this effect was not observed in later passage cells (P7-P15). The cells' ability to utilize LDL-derived cholesterol and to respond to db cAMP (1mM) by increased steroid release was preserved in cells cultured for over 60 generations in the serum-free medium. HDL, although also able to increase steroid production in early-passage cultures exposed to ACTH or to ACTH and dibutyryl cyclic AMP (db cAMP), was 10 fold less potent than LDL. It did not support steroidogenesis in cultures not exposed to these trophic agents. The life span of bovine adrenal cortex cells grown in the serum-free medium on fibronectin (FN)- versus ECM coated dishes was compared. Cells seeded in serum-containing medium and grown in serum-free medium had a life span of 34 versus 60 generations when maintained on fibronectin- or ECM-coated dishes, respectively. Cells seeded in the complete absence of serum in the serum-free medium on ECM- or fibronectin-coated dishes could be passaged for 26 or 13 generations, respectively. While FGF was an absolute requirement for cells cultured on fibronectin-coated dishes, it was not required when cells were maintained on ECM. These observations demonstrate the influence of the ECM not only in promoting cell growth and differentiation but also on the life span of cultured cells. PMID- 6294126 TI - Formyl peptide stimulation of superoxide anion release from lung macrophages: sodium and potassium involvement. AB - We examined the role of the monovalent cations Na+ and K+ in the events encompassing the release of O-2 by alveolar macrophages after stimulation with formyl methionyl phenylalanine (FMP). This was accomplished by determining the effect of changing the extracellular [Na+] and/or [K+] on FMP-stimulated O-2 production; and measuring 22Na+, 42K+ and 86Rb+ influx and efflux and intracellular [K+] for control and FMP-stimulated alveolar macrophages. Stimulated O-2 production was relatively insensitive to changes in extracellular K+ or Na+ concentrations until the [Na+] was decreased below 35 mM. At 4 mM [Na+], the rate of O-2 production remained at 75% of the maximal rate observed at physiological concentrations of [Na+]. Both influx and efflux of 22Na+ were stimulated above control rates by FMP. The increased rates of fluxes lasted for a few minutes suggesting a transient increase in membrane permeability to Na+. Ouabain partially inhibited 22Na+ efflux but had no effect on O-2 release. The influx of 86Rb+ and 42K+ was not altered by the addition of FMP but was virtually abolished in the presence of 10 microM ouabain or 1 mM quinine. In the presence of extracellular calcium, FMP-stimulated a prolonged (greater than 20 minutes) increase in 86Rb+ or 42K+ efflux which was inhibitable by 1 mM quinine. In the absence of extracellular calcium, FMP stimulation of K+ efflux was greatly diminished and was not affected by quinine, although quinine still inhibited O-2 production under these conditions. It was also observed that there was a loss of intracellular K+ when cells were stimulated by FMP in the presence of Ca+2, but not in the absence of Ca+2. Taken together, these results suggest a minimal direct role, if any, for K+ in the events that lead to FMP-stimulated O-2 release by alveolar macrophages. PMID- 6294127 TI - Stimulation of dome formation in MDCK kidney epithelial cultures by inducers of differentiation: dissociation from effects on transepithelial resistance and cyclic AMP levels. AB - Changes in transepithelial electrical resistance and cyclic nucleotide levels were monitored accompanying chemical induction of domes in a clonal subline of MDCK kidney epithelial cells. Confluent cell monolayers grown on nitrocellulose filters exhibited a relatively high mean transepithelial resistance (387 ohms X cm2). Hexamethylene bisacetamide, a potent inducer of dome formation (Lever, 1979b), stimulated significantly increased transmonolayer resistance as well as elevated levels of intracellular cyclic AMP. By contrast, dimethylformamide, an equally potent inducer of dome formation in MDCK cells, did not appreciably alter either resistance values or cyclic nucleotide levels. These results suggest that induction of dome formation in epithelial cell cultures by compounds generally known as inducers of differentiation may involve multiple and separate mechanisms. PMID- 6294129 TI - The effects of temperature and ouabain on steady-state Na and K exchanges in human lymphocytes. PMID- 6294128 TI - Genetic evidence that a phorbol ester tumor promoter stimulates ornithine decarboxylase activity by a pathway that is independent of cyclic AMP-dependent protein kinases in CHO cells. AB - Ornithine decarboxylase (ODC) inductions by cholera toxin and by the phorbol ester tumor promoter, TPA, were compared in wild-type Chinese hamster ovary (CHO) cells and in mutant cells having altered cyclic AMP-dependent protein kinase activity. The aim of these studies was to determine whether cyclic AMP-dependent protein kinase is involved in these inductions. The time course and the magnitude of ODC inductions by either 100 ng/ml cholera toxin or 100 ng/ml TPA were similar in wild-type cells with a maximum at 3-4 hours after treatment and a return to unstimulated levels by 8 hours. Induction of ODC by cholera toxin was suppressed more than 80% in the four protein kinase mutants studied (10215, 10248, 10260, and 10265), strongly implicating a cyclic AMP-dependent kinase step in the mechanism of induction. Similar results were found with the cyclic AMP analog 8 Br-cyclic AMP and the phosphodiesterase inhibitor, methyl-isobutylxanthine. The induction of ODC by TPA, on the other hand, was only partially inhibited (approximately 50%) in three of four mutants. Lower ODC activity in two mutants stimulated by cholera toxin or TPA whose kinetics were studied in more detail could not be ascribed to a reduced affinity (Km) of ornithine for the enzyme, but appeared to be due to reduced catalytic activity (Vmax) in the extracts. These results suggest that the induction of ODC by TPA proceeds by a mechanism which is only partially dependent on an intact cyclic AMP-dependent protein kinase activity. PMID- 6294130 TI - Control of melanin synthesis and secretion by B16/C3 melanoma cells. AB - In culture, B16/C3 murine melanoma cells grown in the presence of serum undergo melanogenesis at a specific time after plating. At this time, melanin is synthesized intracellularly and then secreted into the extracellular culture fluid. We have found that melanin secretion is dependent on the presence of serum in the growth medium. When confluent cultures are deprived of serum, that is, refed with serum-free medium, cells remain viable but do not undergo melanogenesis. Addition of serum-free medium supplemented with either melanocyte stimulating hormone (MSH) or dibutyryl cAMP induced melanogenesis in these cells but did not result in melanin secretion. Furthermore, when B16/C3 cells are grown in serum-free, hormone-supplemented medium, they also undergo melanogenesis but fail to release melanin. The addition of serum, however, to B16/C3 cells induced to undergo melanogenesis with MSH, dibutyryl cAMP, or hormone-supplemented medium promotes melanin secretion. Fractionation studies hence revealed that serum contains specific factors capable of inducing melanin secretion. These results demonstrate that factors that regulate melanin synthesis are distinct from those that induce cells to release melanin into their extracellular environment. Furthermore, the ability to induce melanogenesis with single factors will permit us to study the precise sequence of events leading to differentiation in B16/C3 cells under chemically defined conditions. PMID- 6294131 TI - Growth stimulation of rous sarcoma virus-transformed BHK cells by biotin and serum lipids. AB - Biotin or a serum lipid extract stimulated proliferation of G1 arrested Rous sarcoma virus-transformed BHK cells in modified Eagle's MEM (BM). The cells could be maintained continuously in BM plus biotin (BMB), but not in BM plus serum lipid extract (BM X L). Avidin inhibited growth stimulation when added to BMB, but did not inhibit growth when added to BM X L. 14C-acetate incorporation into total cellular lipids was stimulated in BMB, but not in BM. Thin-layer chromatography of the labeled cellular lipid extract indicated that relatively large amounts of 14C-acetate were incorporated into phosphatidylserine and little into the other major phospholipids. In the neutral lipids, the largest amount of incorporation was in cholesterol. G1 arrested cells multiplied rapidly in BM supplemented with dialyzed serum (BM X DS), but they did not multiply in BM with delipidized serum (BM X DLS). The addition of biotin or serum lipid extract to BM X DLS stimulated growth. Growth stimulation in BM X DLS by biotin was inhibited by avidin, but avidin had no effect on growth stimulation by serum lipid extract. Biotin stimulated additional multiplication in BM X DS and avidin inhibited this additional growth stimulation. These results suggest that growth stimulation requires lipids supplied by serum lipids or by de novo synthesis stimulated by biotin. In the absence of serum, the stimulation of the synthesis of growth factor(s) by biotin are also required for continuous multiplication. PMID- 6294132 TI - Cross-reactivity of antibodies against synthetic peptides. AB - Antiserum against the synthetic peptide Lys-Arg-Ser-Arg-His-Phe, corresponding to the carboxy terminus of polyoma virus medium tumor antigen (medium T antigen), immunoprecipitates a protein of 36,000 daltons from polyoma virus-infected and uninfected cell extracts treated with the sulfhydryl group reagent N-ethyl maleimide. This protein appears to share an antigenic determinant with medium T antigen that is normally buried inside the protein or covered up by another protein or cellular structure. The two-dimensional tryptic fingerprints of the 36K protein and of medium T antigen are apparently unrelated to each other. Antiserum against the octapeptide Ac-Met-Asp-Lys-Val-Leu-Asn-Arg-Tyr, including the amino-terminal heptapeptide sequence of the simian virus 40 (SV40) large tumor (T) and small T antigens, cross-reacts with polyoma virus large T antigen, which has an identical amino-terminal heptapeptide sequence except that Lys is replaced by Arg and Asn by Ser. The problem of cross-reactivities of antipeptide sera is discussed. PMID- 6294133 TI - Detection of a complex of SV40 large tumor antigen and 53K cellular protein on the surface of SV40-transformed mouse cells. AB - The possible interaction between simian virus 40 (SV40) large tumor antigen (T ag) and cellular proteins in the plasma membrane of SV40-transformed mouse cells was investigated. The presence of SV40 T-ag, 53,000 (53K) cellular protein, and histocompatibility (H-2) antigens on the surface of SV40-transformed cells was demonstrated by immunofluorescence. The use of lactoperoxidase-catalyzed cell surface iodination and a differential immunoprecipitation technique established that large T-ag is associated with the 53K host-coded protein on the surface of the transformed cells. In contrast, no detergent-stable complex between large t ag and H-2 antigens was detected. Both labeled T-ag and 53K protein were coprecipitated from surface-iodinated SV40-transformed cells by monoclonal antibodies directed against either the viral or the cellular protein. Based on the unique antigenic sites recognized by the anti-T monoclonal antibodies, it appears that both the carboxy and amino termini of the T-ag polypeptide are exposed on the surface of SV40-transformed mouse cells. The nature of the association between surface T-ag and 53K protein, as well as that between the molecular complex and the plasma membrane, remains to be determined. The possible effect of the surface-associated T-ag/53K complex on cellular proliferation is considered. PMID- 6294134 TI - [Shoulder movements after mammectomy with conservation of muscle. Report of 100 cases]. AB - One hundred patients were treated by a Patey type conservative radical mammectomy with or without associated radiotherapy. Objective numerically rated comparison of the frequency and degree of limitation of the different shoulder movements after surgical treatment alone showed that abduction was the movement most markedly and frequently limited. This was followed, in decreasing order of frequency and degree, by limitation of amplitude, anterior flexion, external rotation, posterior flexion, and internal rotation. In cases with associated radiotherapy, highest percentages of maximal imitation for each movement occurred in those undergoing pre-operative cobalt therapy, and particularly involved abduction, posterior flexion, and external rotation. Patey type mammectomy provides improved functional results when compared with the classical Halsted's operation, but its technique has still to be perfected, a useful basis for this being a detailed analysis of its possible sequelae. PMID- 6294135 TI - Some aspects of cellular aging. AB - Studies were carried out on cultured human fibroblasts in order to elucidate the biology of aging and the origins of age-dependent diseases. The replicative life span of cultures was inversely proportional to the chronological age of the tissue donor, and cultures derived from subjects with two inherited disorders of premature aging, progeria and Werner syndrome, had more severely impaired growth capacity. Studies on circular outgrowths whereby cell division is restricted to a circumferential rim of cells indicated that the replicative life span is controlled by a mitotic counter to a critical limit. The response of progeria cells to a hormone preparation with insulin-like activity was decreased, while in normal cells this decrease occurred as a function of passage level with a "shift to the right" of the dose-response curve. Cyclic AMP content of fibroblasts at late passage fell in response to PGE1 stimulation but rose in response to epinephrine, likely due to altered expression of genes for the receptors for each of these two hormones. This system of cultured human fibroblasts is useful in explaining various concomitants of biological aging including decreased tissue cellularity and impaired hormone and drug responsiveness. PMID- 6294136 TI - Cerebrospinal fluid IgG bands and virus-specific IgG, IgM, and IgA antibodies in herpes simplex virus encephalitis. AB - To characterize the immune response of the central nervous system in herpes simplex virus (HSV) encephalitis, cerebrospinal fluid (CSF) specimens of 7 biopsy proven and 7 presumptive herpes simplex virus (HSV) encephalitis patients were studied, using sodium dodecyl sulfate polyacrylamide gel electrophoresis for the presence of CSF IgG bands, and solid-phase enzyme immunoassays for HSV-specific antibodies. IgG bands were detected in all CSF specimens of the patients, as early as day 6 and up to day 1088. A novel, unidentified, 120 000 dalton polypeptide was found in the CSF of most of the patients, in a total of 25/50 specimens, but not in the controls. This polypeptide was evident by day 6, its intensity fluctuated and it was present in specimens collected as late as day 855. HSV-specific antibodies, of either IgG, IgM, or IgA class, were not detected in the CSF during the first week of illness. IgG antibodies appeared later in all patients and persisted to the end of the follow-up of 3 years. The fact that CSF IgG bands were present in some patients before the appearance of HSV antibodies, and also persisted longer, suggests that the IgG response is not restricted to HSV-specific antigenic determinants. PMID- 6294137 TI - Role of apolipoprotein E-containing lipoproteins in abetalipoproteinemia. AB - Detailed studies of apolipoprotein E (apoE)-containing lipoproteins in abetalipoproteinemia have been performed in an attempt to resolve the apparent paradox of a suppressed low density lipoprotein (LDL) receptor pathway in the absence of apoB-containing lipoproteins. It was hypothesized that apoE-containing high density lipoproteins (HDL) in abetalipoproteinemia might functionally substitute for LDL in regulation of cholesterol metabolism in these patients. The mean (+/-standard deviation) plasma concentration of apoE in nine patients with abetalipoproteinemia was 44.8+/-8.2 mug/ml, slightly higher than the corresponding value for a group of 50 normal volunteers, 36.3+/-11 mug/ml. Fractionation of plasma lipoproteins by agarose column chromatography or by ultracentrifugation indicated that in abetalipoproteinemia, plasma apoE was restricted to a subfraction of HDL. This was in contrast to the results obtained with plasma from 30 normal volunteers, in whom apoE was distributed between very low density lipoproteins (VLDL) and HDL. Consequently, the mean apoE content of HDL in abetalipoproteinemia (44.8 mug/ml) was more than twice that found in the normal volunteers (20.3 mug/ml).ApoE-rich and apoE-poor subfractions of HDL(2) were isolated by heparin-agarose affinity chromatography. ApoE comprised a mean of 81% of the protein mass of the apoE-rich subfraction. Compared with the apoE poor subfraction, the apoE-rich HDL(2) was of larger mean particle diameter (141+/-7 vs. 115+/-15 A) and had a higher ratio of total cholesterol/protein (1.01+/-0.11 vs. 0.63+/-0.14). Plasma and HDL fractions from three patients were studied with respect to their ability to compete with (125)I-LDL in specific binding to receptors on cultured human fibroblasts. The binding activity of plasma from patients (per milligram of protein) was about half that of plasma from normal volunteers. All binding activity in the patients' plasma was found to reside in the HDL fraction. The binding activity of the patients' HDL (on a total protein basis) was intermediate between that of normal HDL and normal LDL. However, the large differences in binding between patients' HDL and normal HDL entirely disappeared when data were expressed in terms of the apoE content of these lipoproteins. This suggested that the binding activity was restricted to that subfraction of HDL particles that contain apoE. These apoE-rich HDL particles had calculated binding potencies per milligram of protein 10-25 times that of normal LDL. Direct binding studies using (125)I-apoE-rich HDL(2) and (125)I-apoE-poor HDL(2), confirmed the suggestion that binding is restricted to the subfraction of HDL particles containing apoE. The apoE-rich HDL(2) were found to be very potent inhibitors of 3-hydroxy-3-methyl-glutaryl coenzyme A reductase activity in cultured fibroblasts, providing direct evidence of the ability of these lipoproteins to regulate cholesterol metabolism. On the basis of binding potencies of apoE-rich HDL, apoE concentrations, and the composition of apoE-rich HDL, it could be calculated that apoE-rich HDL in abetalipoproteinemia have a capacity to deliver cholesterol to tissues via the LDL receptor pathway equivalent to an LDL concentration of 50-150 mg/dl of cholesterol. Thus, these apoE-rich lipoproteins are capable of producing the suppression of cholesterol synthesis and LDL receptor activity previously observed in abetalipoproteinemia. PMID- 6294138 TI - Spontaneous oxygen radical generation by sickle erythrocytes. AB - Since the various membrane abnormalities of sickle erythrocytes might result from excessive accumulation of oxidant damage, we have measured the generation of superoxide, peroxide, and hydroxyl radical by normal and sickle erythrocytes using assays involving reduction of cytochrome c, aminotriazole inhibition of catalase, and methane evolution from dimethyl sulfoxide, respectively. Compared with normal erythrocytes, sickle erythrocytes spontaneously generate approximately twice as much superoxide, peroxide, and hydroxyl radical. One possible source of hydroxyl radical generation was identified as hemichrome, excessive amounts of which are bound to sickle erythrocyte membranes. Hemichrome did not generate hydroxyl radical when exposed to superoxide alone or peroxide alone. However, in the presence of both superoxide and peroxide, hemichrome greatly facilitated hydroxyl radical generation. Supporting this, normal erythrocyte membranes induced to acquire sickle hemichrome concomitantly acquired an enhanced ability to mediate hydroxyl radical generation. Finally, sickle erythrocyte membranes greatly enhanced superoxide/peroxide-driven hydroxyl radical generation as compared with normal erythrocyte membranes. These data suggest that an excessive accumulation of oxidant damage in sickle erythrocyte membranes might contribute to the accelerated membrane senescence of these cells. They further indicate that accumulation of oxidant damage could be a determinant of normal erythrocyte membrane senescence. PMID- 6294139 TI - Evidence of normal functional levels of activated protein C inhibitor in combined Factor V/VIII deficiency disease. AB - Human activated protein C (APC) is a plasma serine protease that possesses amidolytic and anticoagulant activity. The rate at which the amidolytic and anticoagulant activity of APC was neutralized in normal plasma was essentially identical to that observed in plasma obtained from four individuals with combined Factor V/VIII deficiency disease. Incubation of radioiodinated APC with either normal human plasma or the combined Factor V/VIII-deficient plasmas resulted in the formation of a stable complex (Mr = 96,000) of the enzyme and a plasma protein as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Pretreatment of the radiolabeled APC with diisopropyl fluorophosphate prevented the formation of the enzyme-protein complex. On the basis of its ability to form a complex with radiolabeled APC, the APC-binding protein was purified to homogeneity from normal human plasma by ammonium sulfate fractionation, heparin-agarose chromatography, and QAE-Sephadex A-50 chromatography. The APC-binding protein (Mr = 54,000) is a glycoprotein, and possesses an amino-terminal sequence of Gly-Arg-Thr-Cys-Pro-Lys-Pro-Asp. The amino-terminal sequence of the APC-binding protein exhibited considerable homology with bovine colostrum inhibitor and pancreatic trypsin inhibitor, but no apparent sequence homology with the plasma serine protease inhibitors. Affinity purified antibody against APC-binding protein immunoprecipitated a complex of radiolabeled APC and native APC-binding protein from normal human plasma. Complex formation was virtually eliminated in plasma immunodepleted of the APC-binding protein. Quantitative electroimmunoassay indicated essentially equal levels of APC-binding protein antigen in normal plasma compared with plasma from four patients with combined Factor V/VIII deficiency disease. PMID- 6294140 TI - Beta endorphin selectively stimulates aldosterone secretion in hypophysectomized, nephrectomized dogs. AB - We examined the effects of synthetic human beta-endorphin (beta END) and a stable methionine (Met)-enkephalin analogue on aldosterone and cortisol secretion rates in anesthetized, hypophysectomized, and nephrectomized dogs and compared them to those of (1-39) ACTH. The circulation of the adrenal glands was completely isolated on the arterial and venous sides (Hilton Pouch). The peptides were infused to deliver 3 pmol/min into the aortic "pouch." Blood was collected from the vena caval pouch, which received blood only from the adrenal gland. Secretion rates of aldosterone and cortisol were calculated as the product of adrenal blood flow and venous steroid concentration. Duplicate steroid measurements were obtained during a control period, at 10, 30, and 50 min of peptide infusion and during a postcontrol period. BetaEND increased aldosterone secretion rate from 2.4 +/- 0.5 ng/min (mean +/- SEM) to 3.2 +/- 0.9 ng/ min at 10 min (N.S.), 8.2 +/ 2.5 ng/min (P less than 0.05) at 30 min and 11.0 +/- 3.7 ng/ min (P less than 0.05) at 50 min of infusion. Cortisol secretion rate was not affected by infusion of betaEND. Infusion of the stable Met-enkephalin analogue D-alanine2; Metphenylalanine4, Met(O)-enkephalin-ol or saline alone had no effect on aldosterone or cortisol secretion rates. ACTH infusion increased mean aldosterone secretion rate by approximately 215% and significantly stimulated cortisol secretion rate. These results indicate that beta END selectively stimulates aldosterone secretion with a potency similar to that of an equimolar dose of ACTH. PMID- 6294142 TI - Sonographic diagnosis of intraductal hepatoma. PMID- 6294141 TI - Preferential binding of vasoactive intestinal polypeptide to basolateral membrane of rat and rabbit enterocytes. AB - Binding of radioiodinated vasoactive intestinal polypeptide (VIP) to intestinal cell membranes of the rabbit ileum and rat jejunum was investigated. Specific binding of 125I-labeled VIP could be demonstrated only on the basolateral membrane and not on the brush border membrane. This corresponded with the lack of an effect on ion transport when VIP was applied to the mucosal side of an in vitro preparation of rabbit ileum. VIP altered ion transport only when it was applied to the serosal side. The binding of 125I-VIP was specific and dependent upon incubation temperature. There was a close correlation between the potency of VIP for inhibition of 125I-VIP binding and that for increasing adenylate cyclase activity. These observations demonstrate that VIP receptors are located on the basolateral membrane. PMID- 6294143 TI - Sonographic findings in a patient with pseudomyxoma peritonei. PMID- 6294144 TI - Comparison of enzyme-linked immunosorbent assay, radioimmunoassay, complement fixation, anticomplement immunofluorescence and passive haemagglutination techniques for detecting cytomegalovirus IgG antibody. AB - The radioimmunoassay (RIA) and enzyme-linked immunosorbent assay (ELISA) techniques were found to be comparable in sensitivity and specificity for detecting cytomegalovirus IgG antibody, and 10 to 100 times more sensitive than complement-fixation (CF), anticomplement immunofluorescence (ACIF) and passive haemagglutination (PHA). In screening tests for antibody, the frequency of false positive and -negative results was 0.6% for RIA and ELISA, 1.5% for CF, 1.6% for ACIF and 3.6% for PHA. PHA was the least satisfactory test, largely because of technical problems. Cytomegalovirus (CMV) infection is an important cause of congenital brain damage and is also a major complication of both prolonged immunosuppressive therapy, especially in patients with organ transplants, and multi-donor blood transfusions. For serological diagnosis of infection, as well as for screening for antibody in patients and in blood donors, the solid-phase indirect radioimmunoassay (RIA) and enzyme-linked immunosorbent assay (ELISA) techniques offer distinct improvements in sensitivity over previous methods. Although the principle of both tests, based on the detection of antigen-antibody reactions by means of a labelled anti-antibody, is the same, each possesses its own particular technical advantages and disadvantages, and both require their own expensive equipment for the reading of the results. There is still a lack of data on how they compare in sensitivity and specificity. The present study was undertaken to compare the two methods for the detection of CMV IgG and to evaluate them against the older techniques of complement-fixation (CF), passive haemagglutination (PHA) and anticomplement immunofluorescence (ACIF). PMID- 6294145 TI - A simple disc technique for the presumptive identification of Legionella pneumophila. AB - A method is described for the presumptive identification of Legionella pneumophila by the formation of satellite colonies around filter paper discs impregnated with ferric pyrophosphate and L-cysteine hydrochloride on a deficient basal medium. This technique simplifies the differentiation of picked colonies of L pneumophila from other organisms in mixed cultures from environmental and contaminated clinical samples. PMID- 6294146 TI - The potential of bacteriocin typing in the study of Clostridium perfringens food poisoning. AB - A range of 49 bacteriocins was used to type 311 strains of Clostridium perfringens isolated from food poisoning outbreaks. Strains of same serotype within an outbreak showed similar patterns of susceptibility to bacteriocins, whereas strains of different serotype isolated from different sources produced many variations in bacteriocin susceptibility patterns. The 311 strains, along with isolates from a wide range of sources were screened for their ability to produce bacteriocins. A much greater proportion of the strains from food poisoning outbreaks was bacteriocinogenic than were isolates from human and animal infections, various foods and the environment. PMID- 6294147 TI - Alveolar-cell carcinoma: a problem in sputum cytodiagnosis. AB - Cytology and histology are correlated in a series of 22 cases chosen to illustrate the differential diagnosis between clusters of benign bronchial or bronchiolar cells seen in sputum, and those of alveolar cell carcinoma or adenocarcinoma with alveolar spread. Alveolar-cell carcinoma is characterised by clusters of small epithelial cells in spherical or irregular formations, none showing enough polarity to distinguish a smooth or palisaded surface. The appearances are most distinctive if vacuolation is absent. The diagnosis cannot, however, be confidently made in all cases from morphological features of cells in sputum. PMID- 6294148 TI - A comparative histological and electrophysiological study of some neurotoxins in the rat hippocampus. PMID- 6294149 TI - Functional role of regrowing pyramidal tract fibers. AB - When pyramidal tract axons are severed in the infant hamster, the damaged fibers regrow via a new pathway to their normal terminal sites in the medulla and spinal cord and there form synaptic connections (Kalil and Reh, '79, '82). We studied the behavior of animals with infant and adult lesions of the medullary pyramid to determine the functional significance of the new pathway in maintaining normal motor behavior. Examination of behaviors normally mediated by the pyramidal tract, particularly the manipulation of sunflower seeds during feeding, revealed a correlation between the presence of the new tract and the preservation of function. Furthermore, in the adult animal with an infant lesion, the spared behaviors were lost when the new pathway was destroyed. PMID- 6294150 TI - An intracellular HRP study of the rat globus pallidus. I. Responses and light microscopic analysis. AB - A study of the intracellularly recorded responses of rat globus pallidus neurons to activation of striopallidal fibers was combined with light microscopic examination of the morphology of these same neurons using intracellular horseradish peroxidase. The response to stimulation of caudate-putamen is an inhibitory postsynaptic potential with observed latencies ranging from 5.1 to 9.8 msec. These values correspond to conduction velocities of 0.4 to 0.8 m/second for striopallidal fibers. Comparison with extracellular controls shows no excitatory component to the response. All recovered and analyzed neurons (n = 11) were of the large type of pallidal neuron known from Golgi studies but the addition that two subtypes could be recognized. Large neurons located medially in the nucleus had dendritic fields with large dorsoventral extent (ca. 1 mm) when compared to their mediolateral and rostrocaudal dimensions (ca. 0.4 mm) and these neurons emitted no axon collaterals. Large neurons located laterally in the nucleus had disklike dendritic fields with both dorsoventral and rostrocaudal dimensions being on the order of 1 mm but with a minor axis of approximately 100 micrometers. The axons of these neurons possessed collaterals. As a consequence of their disk-shaped dendritic field, neurons belonging to the laterally placed subgroup and occupying the narrow (ca. 100 micrometers thick) striopallidal border zone known to receive a distinct input from neostriatum have dendrites restricted to that zone. PMID- 6294151 TI - The motor cortex of the rat: cytoarchitecture and microstimulation mapping. AB - The first motor (MI) cortex of the rat was identified as the region from which movements could be evoked by the lowest intensity of electrical stimulation. The location of this region was correlated with cytoarchitecture in the frontal and parietal cortex. Two frontal areas can be discerned in Nissl-stained sections: (1) the medial agranular field, marked by a pale-staining layer III and a compact layer II, and (2) the lateral agranular field, which has more homogeneous superficial layers and a broad layer V containing large, densely staining cells. Both of these regions project to the spinal cord and can therefore be included in the somatic sensorimotor cortex. MI in the rat coincides with the lateral agranular field but also overlaps with part of the adjacent granular cortex of the first somatic sensory (SI) representation. We conclude that the rat MI cortex can be identified by microstimulation techniques and by cytoarchitecture in the rat. PMID- 6294152 TI - Genetic predisposition to Kaposi's sarcoma. PMID- 6294153 TI - Brachial plexus: anatomic, radiologic, and pathologic correlation using computed tomography. AB - Computed tomographic (CT) anatomy of the normal brachial plexus was analyzed in 50 patients with nonneurologic disease. Identification of anatomic structures on the CT images was facilitated by comparison with cadaver cross sections. Ten patients with clinical brachial plexopathy were also examined. Of these 10 patients, 7 demonstrated lesions of the brachial plexus. Computed tomographic examination of the brachial plexus is useful in detecting mass lesions of the plexus and may allow earlier diagnosis and specific therapy. Whether CT will be reliable in distinguishing idiopathic brachial plexopathy from the plexopathy associated with pathological lesions awaits further study. PMID- 6294154 TI - CT detection of hepatic metastases with Ethiodized Oil Emulsion 13. AB - The use of an experimental liposoluble contrast agent-Ethiodized Oil Emulsion 13 (EOE 13)--is described in hepatic computed tomography (CT) of 23 oncologic patients. Without exception, all the hepatic metastases were better delineated in the EOE 13 enhanced scans. The postcontrast scans also detected an increased number of lesions but not all were malignant. There is no specificity to the increased lesion detection. Various splenic abnormalities were detected. Very few minor side effects and no major side effects were caused by the contrast media. We feel that with time, hepatic specific agents such as EOE 13 will become the contrast media of choice in hepatic CT examinations. Also, CT with hepatic specific agents may become the preoperative examination of choice in candidates for partial hepatectomy. PMID- 6294155 TI - Biodistribution study of Ethiodized Oil Emulsion 13 for computed tomography of the liver and spleen. AB - Biodistribution studies were conducted with a new intravenous lipoid contrast material currently undergoing clinical trials in four hospitals. The contrast material selectively opacifies the liver and spleen for computed tomographic examination. The experiments were performed on rats with 125I-labeled ethiodized oil emulsion. The study showed that the liver accumulates nearly 80% of the injected iodine within 15 min of the injection and retains a high concentration over 3 h. The second highest concentration was found in the spleen. More than 99% of the iodine is eliminated from the liver and spleen within 48 h, primarily through the kidneys. PMID- 6294156 TI - Mucinous adenocarcinoma of the prostate. PMID- 6294157 TI - Modulation of catecholamine activation of adenylate cyclase by the number of active beta-adrenergic receptors: theoretical considerations on the role of receptor diffusion in the cell membrane. AB - Numerous experimental studies on the beta-adrenergic receptor suggested that the adenylate cyclase activity is correlated to the number of agonist-receptor complexes. In S49 lymphoma cells, it has been previously shown that, for a given number of occupied receptors, the level of adenylate cyclase activity depends on the total number of receptors. Here we propose and demonstrate that such a behaviour can be accounted for by the cyclic model of Cassel and Selinger provided the diffusion of the receptor in the plane of the membrane is explicitly introduced in the model. The key properties of this expanded version of the model are related to the relative kinetics of receptor diffusion and of agonist dissociation from the receptor. PMID- 6294158 TI - beta-Adrenergic receptors and cyclic AMP responses to epinephrine in cultured human fibroblasts at various population densities. AB - The beta-adrenergic receptors of the intact human lung diploid fibroblast line Wl 38 and an SV-40 transformed clone of Wl-38, Wl-38-VA-13-2RA (VA13), were estimated in experiments utilizing the beta-adrenergic ligand, 125l hydroxybenzylpindolol (125IHYP). When specific 125IHYP binding was measured in cells grown to relatively low population densities (0.15x10(6)cells/35mm dish), both Wl-38 and VA13 cells had approximately 40,000 beta-adrenergic receptors per cell. Wl-38 cells, when cultured to a high population density (0.5x10(6) cells/35/mm dish) had clearly diminished numbers of beta-adrenergic receptors and greatly decreased cAMP responses to epinephrine stimulation. On the other hand, in VA13 cells, neither the receptor number nor the beta-adrenergic response was affected by cell population density. In Wl-38 cells, the diminished cAMP response to epinephrine paralleled the decrease in number of beta-adrenergic receptors. Prostaglandin E1 (PGE1) stimulation of cAMP levels was unaffected by cell population density in either Wl-38 or VA13 cells. Thus, increased cell population density, perhaps related to density dependent inhibition of growth, caused a specific diminution in 125IHYP binding concomitant with decreased cAMP responses to epinephrine. PMID- 6294159 TI - Influence of sodium bicarbonate on growth and health of young calves. AB - Fifty-four Holstein and Jersey calves were assigned at 4 days of age within breed and sex to one of four treatments: control consisting of colostrum, milk replacer, and starter; buffered colostrum and replacer (.6% sodium bicarbonate) and starter (2% sodium bicarbonate); acidified colostrum (1% propionic), untreated replacer, and starter; and acidified, buffered colostrum (1% propionic, .6% sodium bicarbonate), buffered replacer (.6% sodium bicarbonate), and starter (2% sodium bicarbonate). The feeding regimen was colostrum once daily, day 4 to 14; milk replacer once daily, day 15 to 28; and calf starter ad libitum, day 4 to 84. Bull calves were fed for 42 days and heifers for 84 days. Calves fed acidified colostrum refused more feed and were less efficient from day 4 to 14 than calves fed buffered colostrum. Bulls were more sensitive to acidified colostrum than heifers. Starter intake, total dry matter intake, and average daily gains were similar for all calves during days 4 to 84. Rumen fluid from calves fed diets with sodium bicarbonate was higher in acetate and lower in propionate and lactate than that from calves fed diets without sodium bicarbonate. Sodium bicarbonate improved intake of acidified colostrum during the first 2 or 3 days of feeding but had no other effect on gain or feed intake. PMID- 6294160 TI - Solvent effects on bonding organo-silane to silica surfaces. AB - Interfacial bonding and stability of gamma-methacryloxypropyltrimethoxysilane with silica surfaces have been studied by means of infrared spectroscopy. The addition of n-propylamine enhances silanization of gamma methacryloxypropyltrimethoxysilane to silica surfaces in normal aliphatic hydrocarbons, and cyclohexane yields a more water-resistant silica-silane bond, and improves the diametral tensile strength of the composite. PMID- 6294161 TI - Extraskeletal implantation of a porous hydroxyapatite ceramic. AB - Coralline replamineform hydroxyapatite implants were placed in canine subcutaneous tissues in an attempt to clarify whether the hydroxyapatite matrix acted as a passive matrix for osseous ingrowth when placed in some inherent bone induction capacity. The implants were well-tolerated and elicited no deleterious host response. Connective tissue rapidly infiltrated the pores, but no evidence of bone formation was noted in any of the specimens. It was concluded that this implant material does not act to induce bone formation. PMID- 6294162 TI - Synovial sarcoma of the foot. PMID- 6294163 TI - Dermabrasion for the management of angiofibromas in tuberous sclerosis. PMID- 6294164 TI - Dietary fiber content of some tropical fruits and vegetables. PMID- 6294165 TI - Nutritional evaluation of seashore mallow seed, Kosteletzkya virginica. PMID- 6294166 TI - Evidence for adrenergic modulation of ganglionic transmission in the ganglia of the nerve of Remak of the chicken. AB - Adrenergic depression of ganglionic transmission in non-cholinergic, non adrenergic (NCNA) excitatory nerves was investigated in the isolated chicken rectum with its associated nerve of Remak. Preganglionic stimulation of the NCNA nerves produced contraction of the rectum, an excitatory junction potential and discharges in the postganglionic axons. Preceding stimulation of descending fibers in the nerve of Remak suppressed these responses. These inhibitory effects were almost eliminated in reserpinized preparations, and were significantly reduced by pretreatment with guanethidine and phentolamine, but not with propranolol. Exogenously-applied adrenaline and noradrenaline mimicked the inhibitory effect of nerve stimulation on the discharge in the postganglionic axons. These results indicated that the nerve of Remak contained descending adrenergic fibers, stimulation of which caused suppression of ganglionic transmission in the NCNA excitatory pathways to the rectum, by the activation of alpha-adrenoceptors on the pre- or postganglionic NCNA neuron. PMID- 6294167 TI - Some characteristics of transmission from non-adrenergic, non-cholinergic excitatory nerves to the smooth muscle of the chicken. AB - Non-adrenergic, non-cholinergic excitatory junction potentials (e.j.p.s) were recorded from the longitudinal muscle cells of the chicken rectum following stimulation of the nerve of Remak and field stimulation of intramural nerves. The amplitude of e.j.p.s was a direct function of the stimulus intensity. When peak depolarization of an e.j.p. reached threshold, a muscle action potential was evoked. When the extrinsic nerve was stimulated with a given stimulus intensity, the e.j.p.s recorded from different cells showed little change in amplitude and time-course, reached their peaks in 74 ms and declined to half their amplitudes in 230 ms. These corresponding values for the e.j.p.s in response to field stimulation were 51 ms and 159 ms respectively, but became larger as the microelectrode was moved away from the stimulating site. The amplitude became smaller also and the distance at which the e.j.p. declined to half its control amplitude was 4.3 mm. The time constant for the exponential decay phase of the e.j.p. was the same as the time constant for the muscle membrane. Facilitation of e.j.p. amplitude occurred during repetitive nerve stimulation; however, the facilitation was accompanied frequently by an initial depression phase during extrinsic nerve stimulation. PMID- 6294168 TI - Involvement of both adrenergic and cholinergic receptors in the cardiovascular effects of naloxone during hemorrhagic hypotension in the conscious rabbit. AB - Opiate receptor blockade with naloxone reverses the hypotension associated with severe hemorrhage in a variety of animal models. In the present study, we examined the mechanisms of naloxone's actions in conscious rabbits made hypotensive by hemorrhage. This was accomplished through pharmacological blockade of the efferent limbs of the sympathetic or parasympathetic nervous systems prior to naloxone injection. In addition, we examined the effects of naltrexone in the same model. Naloxone treatment in hypotensive-hypovolemic, conscious rabbits results in an increase in mean arterial blood pressure (BP) and a decrease in heart rate (HR). The bradycardia appears to be due to a reduction in beta adrenergic and an increase in muscarinic-cholinergic activity. The pressor effect is apparently due to increased alpha-adrenergic receptor activation, and is accompanied by an increase in cardiac output, stroke volume, and total peripheral resistance. Naltrexone did not significantly affect BP but it did reduce HR. The results from the present study suggest that naloxone's effects are mediated by an integrated response of the sympathetic and parasympathetic nervous systems. The actions of naloxone may be mediated through antagonism of endogenous opiates. PMID- 6294169 TI - Acidification of a Solonetzic soil by nitrogenous fertilizers. AB - Annual applications of (NH4)2SO4, NH4NO3 and urea on a Solonetzic soil at 112 kg N/ha for 10 consecutive years reduced pH levels from 5.6 for the check to 4.4, 4.9 and 5.3, respectively for (NH4)2SO4, NH4NO3 and urea. (NH4)2SO4 generated twice as much exchange acidity as NH4NO3 and four times as much as urea. Net extractable cations leached from the Ap horizon closely approximated the amount of exchange acidity generated by (NH4)2SO4 and NH4NO3 fertilizers. The levels of soil extractable Al and Mn were greatly enhanced by (NH4)2SO4 as were plant contents. Similar acidifying effects to that produced by the (NH4)2SO4 occurred when NH4NO3 was applied at 300 kg N/ha annually for 12 consecutive years in another field experiment on the same soil. Liming samples of the field (NH4)2SO4 acidified soils in the greenhouse, significantly increased yields and lowered the Al and Mn contents of the plants to normal levels. PMID- 6294171 TI - Plasma cortisol response to ACTH does not accurately indicate the state of hypothalamic-pituitary-adrenal axis. AB - The hypothalamic-pituitary-adrenal function was studied in 55 patients with various pituitary disorders. In particular, the consistency between the responses of plasma cortisol to exogenous ACTH and to insulin hypoglycemia was investigated in 5 patients in whom cortisol response to insulin was absent; four of these patients showed a cortisol response to ACTH of variable degree. These 4 patients had surgical or functional hypothalamus-pituitary disconnection and showed a preserved cortisol response to lysine vasopressin. These data demonstrate the unreliability of ACTH test in assessing hypothalamic-pituitary-adrenal function in hypopituitary patients. PMID- 6294172 TI - Effect of age on red cell membrane sodium -potassium dependent adenosine triphosphatase (Na+-K+ ATPase) activity in healthy men. AB - Decreased cellular thermogenesis may represent a normal aspect of the aging process. Whereas Na+-K+ ATPase appears to be involved directly in body metabolism, enzyme activity on the erythrocyte membrane Na+-K+ ATPase was significantly lower in aged men. In a separate series of experiments, each with representation from the two age groups, red cell membrane Na+-K+ ATPase was also found to be decreased significantly in the aged men. Although mean values were statistically different, one of the elderly men had enzyme activity above that of the mean of the younger men. None of the younger men had enzyme activity below the mean of the older group. Red cell membrane Na+-,+ ATPase activity may be an indicator of physiological aging. PMID- 6294170 TI - Primary hypothyroidism associated with secondary adrenocortical insufficiency. AB - A 50 year old man is described in whom primary hypothyroidism and isolated ACTH deficiency leading to adrenocortical insufficiency occurred simultaneously. Low thyroid hormone levels, elevated serum TSH values and high titers of antithyroid antibodies (ATA) were consistent with primary hypothyroidism. The diagnosis of secondary adrenocortical insufficiency was based on low cortisol and ACTH morning levels, low urinary steroids, a significant increase of cortisol levels after 0.25 mg tetracosactide iv, an absence of ACTH and cortisol release after insulin induced hypoglycemia and a similar absence of ACTH and 11-deoxycortisol increase after metyrapone administration. After one year treatment and five days withdrawal of cortisol administration, the insulin-induced hypoglycemia still failed to elicit secretory responses of ACTH and cortisol, despite the normalization of thyroid hormone levels. The etiopathogenesis of these two coexisting endocrine deficiencies is discussed. PMID- 6294173 TI - Ascorbate radical and ascorbic acid level in human serum and age. AB - Free radical in human serum was observed by an electron spin resonance (ESR) technique and was assigned to ascorbate radical. Quantitative estimation revealed that the ESR intensity could be used as an expedient mean. The ESR intensity of ascorbate radical, the concentrations of total ascorbic acid and oxidized form of ascorbic acid were determined on sera of 200 healthy individuals whose ages ranged from 12 to 96 years. The ESR intensity as well as the concentrations of total and oxidized form of ascorbic acid declined with age. There were significant correlations between the ESR intensity and the concentrations of total and oxidized form of ascorbic acid. From these results, the clinical significance of the concentrations of total ascorbic acid, ascorbate radical and the oxidized form was discussed in relation to age. PMID- 6294174 TI - Congenital anomalies of the upper limb among the Chinese population in Hong Kong. AB - Three hundred twenty-six patients with 396 congenital upper limb anomalies are classified according to the system adopted by the International Federation of Societies for Surgery of the Hands. The relative incidence of each separate anomaly is listed and compared with that of other series. Two hundred cases are also analyzed with respect to the family history, pregnancy history, neonatal history, and associated anomalies and the results are discussed. Significant racial differences were found in the incidence of polydactyly, syndactyly, anomalies associated with syndromes, and transverse arrests. The incidence of polydactyly is higher in this series than in comparable western series. PMID- 6294175 TI - The use of ranitidine in the management of duodenal ulcer: controlled and open comparison with cimetidine in 59 patients. AB - Forty patients with duodenal ulcer were randomly allocated either ranitidine 150 mg twice daily or cimetidine 1 g daily on a single blind basis for 4 weeks initially, with an additional month of treatment if endoscopy showed incomplete healing. The endoscopist was unaware of the patients' treatment. After 8 weeks treatment, the healing rate was 85% for ranitidine and 95% for cimetidine; the difference was not statistically significant. An additional 19 patients who did not fulfill the trial criteria, 11 of whom had not responded to cimetidine, were openly treated with ranitidine 150 mg twice daily. In 16 of the patients, the ulcer had healed at 8 weeks (84%), but in one patient, the ulcer perforated after 6 weeks of therapy. Thus, although overall healing rates with ranitidine and cimetidine are similar, ranitidine may be useful in patients where cimetidine has proved ineffective. PMID- 6294176 TI - Investigation on an outbreak of Japanese encephalitis in Raipur, Madhya Pradesh. PMID- 6294177 TI - A monoclonal antibody reactive with Marek's disease tumor-associated surface antigen. AB - An antibody-secreting hybridoma, RPH-6, to Marek's disease tumor-associated surface antigen (MATSA) was produced by somatic-cell hybridization between the mouse myeloma SP2/O-Ag/14 and spleen cells from MSB1 immunized mice. The antibody reacted with 95 to 100% of the cells from eight of 10 chicken MD cell lines and one of two turkey MD cell lines. It did not react with chicken MD cell lines RP1 and SK3, turkey MD cell line RP19, lymphoid leukosis (LL) cell lines, RP9 and RP12, and reticuloendotheliosis virus (REV) cell line RP14. A weak reaction was observed with REV cell line RP13 (10 to 20%), and a very slight reaction with normal chicken spleen cells (1 to 5%). RPH-6 produces immunoglobulin of the IgM class. Cell culture and mouse ascitic fluids have titers of 10(2) and 10(6), respectively, by fluorescent antibody (FA) test against MD tumor cell lines. The antibody did not react with Marek's disease virus (MDV) internal antigen or membrane antigen as detected by indirect FA test on chicken embryo fibroblast cultures grown on coverslips. The 51Cr release assay showed RPH-6 is highly cytotoxic (60% release) only against MD lymphoblastoid cell lines, with antibody prepared from mouse ascites having a cytotoxic titer approaching 10(6). Results from using RPH-6 for differential diagnosis of chicken lymphoid tumors of unknown origin were in complete agreement with those obtained with rabbit anti-MATSA reference serum and pathologic diagnosis. PMID- 6294178 TI - Regulation of neutrophil inflammatory mediator release: chemotactic peptide activation of stimulus-dependent cytotoxicity. AB - Human neutrophils stimulated with phorbol myristate acetate (PMA) were able to damage human erythroleukemic K-562 target cells as assessed by a 3-hr 51Cr release assay. Neutrophils from a patient with chronic granulomatous disease of childhood were ineffective in mediating PMA-stimulated cytolysis. Cytotoxicity was inhibited under anaerobic conditions as well as by catalase and several free radical scavengers. Superoxide dismutase, azide, and cyanide failed to inhibit PMA-dependent cytotoxicity. The influence of the chemotactic peptide N-formyl methionyl-leucyl-phenylalanine (FMLP) on stimulus-dependent cytotoxicity was examined. Neutrophils preincubated with 1.0 X 10(-7) M FMLP did not display an increased cytotoxic response, but were markedly amplified in their ability to effect cytotoxicity with the addition of PMA. Enhancement of PMA-stimulated cytotoxicity resulted from a reversible cellular response to FMLP. The kinetics of the cytotoxic responses reflected the possibility that chemotactic peptide treated neutrophils released cytotoxic mediators to a greater extent and at a faster rate than did untreated neutrophils. Although azide and superoxide dismutase did not inhibit cytotoxic responses of chemotactic peptide-activated neutrophils, the response was prevented by catalase and was markedly inhibited by several free radical scavengers. The ability of FMLP to enhance cytotoxic responses correlated well with its enhancement of PMA-stimulated chemiluminescence under a variety of conditions. In addition, the ability of PMA stimulated neutrophils to mediate methane generation from dimethyl sulfoxide and ethylene generation from alpha keto-gamma-methiol-butyric acid (KMB)--assays that quantitate the generation of oxidizing radicals--was increased if the neutrophils were preincubated with FMLP. These results demonstrate that a chemotactic factor greatly potentiates the release of cytotoxic mediators from neutrophils upon stimulation with a nonchemotactic agent. The cytotoxic mediators appear to be products of oxidative metabolism. The cytotoxic potential of neutrophils that have responded to chemotactic stimuli to reach sites of inflammation may be activated in a similar manner. PMID- 6294179 TI - Granulocyte activation by endotoxin. II. Role of granulocyte adherence, aggregation, and effect of cytochalasin B, and comparison with formylated chemotactic peptide-induced stimulation. AB - PMN stimulation by endotoxin is heavily dependent on incubation conditions: although marked increase of PMN adhesiveness, enzyme release, and hexose monophosphate shunt activity, as well as superoxide production, occur when endotoxin-challenged cells are incubated in stationary petri dishes, absolutely no such response is observed with cells kept in suspension. Preincubation with cytochalasin B does not alter this reaction pattern. Hyperadhesiveness of endotoxin-challenged PMN is retained after thorough washing, eliminating the possibility that adhesion is due to absorption of endotoxin to petri dishes. In contrast to formylated chemotactic peptides that stimulate PMN adherence as well as aggregation, endotoxin elicits absolutely no aggregatory response (+/- cytochalasin B). Preexposure to endotoxin does not block the aggregatory response to subsequent addition of chemotactic peptides, whereas effects of endotoxin and formylated peptides on PMN in the stationary system are additive. These findings indicate that contact to surfaces is an essential primary event in PMN stimulation. PMID- 6294180 TI - Hormonal activation of adenylate cyclase in macrophage membranes is regulated by guanine nucleotides. AB - Many macrophage functions such as chemotaxis, phagocytosis, enzyme secretion, and cytotoxicity are influenced by intracellular cyclic nucleotide levels, but the regulatory mechanisms involved are poorly defined. We have developed methods that allowed us to study the activation of AC in isolated guinea pig (g.p.) macrophage membranes. AC in these membrane preparations could be stimulated approximately twofold by guanine nucleotides. We could not obtain any hormonal activation of membrane-bound AC in the absence of guanine nucleotides. In the presence of GTP, however, the hormones isoproterenol and PGE1 elicited an additional threefold rise in AC activity, which subsided after approximately 15 min. As little as 10( 8) M concentrations of these two hormones induced significant elevations of AC activity. Replacement of GTP by its nonhydrolyzable analogue Gpp(NH)p resulted in a persistent hormone-independent activation of AC, and addition of hormones enhanced this level of activation. Thus, GTP-ase activity is present in macrophage membrane preparations and serves to regulate AC activation. Hormonal stimulation of AC was receptor mediated, because the effect of the beta adrenergic agonist isoproterenol, but not PGE1, was inhibited by the beta adrenergic blocker propranolol. In addition, the potency series of PG corresponded to that observed for stimulation of cAMP production in intact g.p. macrophages, i.e., PGE1 = PGE2 greater than PGA1 greater than PGF2 alpha. AC activation by PG in the membrane preparation was inhibited by an alpha-adrenergic agonist, thus demonstrating one means for down regulating cAMP production in g.p. macrophages. Our studies also showed that certain hormones (e.g., beta-adrenergic agonists, PG) can exert their effect on cAMP production by stimulation of membrane-bound AC, whereas other agents such as lectins or arachidonic acid require additional intracellular components to elevate cAMP levels in macrophages. The mechanism of activation of AC by hormones in g.p. macrophage membranes appears to fit the model of a ternary complex, the components of which include the hormone receptor, AC, and guanine nucleotide regulatory protein, which transmits the signal from the receptor to AC. PMID- 6294182 TI - Natural killer (NK) cells as a responder to interleukin 2 (IL 2). II. IL 2 induced interferon gamma production. AB - In the accompanying paper, we showed that natural killer (NK) cells were a major population in the naive spleens of normal mice that responded directly to a T cell growth factor, interleukin 2 (IL 2), and clonally replicated without other stimulating agents. The cloned cells growing in IL 2 showed a potent NK activity against several NK targets without addition of an NK-activating agent, interferon (IFN). In the present study, therefore, we examined whether these cloned NK cells on their own produced IFN. It was found that all NK clones growing in IL 2 produced IFN in the culture fluids. The titers of IFN produced in the IL 2 containing media correlated well with the number of growing cells. With the culture in the absence of IL 2, neither cell growth nor IFN production could be detected. Addition of Con A into the culture in the IL 2-free media showed no IFN production. The antiserum neutralizing IFN alpha and IFN beta failed to significantly neutralize IFN produced by NK clones. Treatment with either a pH of 2.0 or antiserum neutralizing mouse IFN gamma resulted in a marked reduction of IL 2-induced NK IFN, indicating that a major part of IFN produced was IFN gamma. These results indicate that IL 2 stimulates NK clones to proliferate, accompanied by IFN gamma production. The results also show that an NK clone, when stimulated with Sendai virus, produced a type 1 IFN (IFN alpha and/or IFN beta), suggesting that murine NK cells can produce both type 1 (alpha and/or beta) and type 2 (gamma) IFN, depending on inducers. PMID- 6294181 TI - An unusually high-titer human anti-Epstein Barr virus (EBV) serum and its use in the study of EBV-specific proteins synthesized in vitro and in vivo. AB - Sera from a patient with a chronic Epstein Barr virus (EBV) infection contained unusually high anti-EBV antibody titers (1:2560 to 1:10,240 for EA(D) and 1:5,120 to 1:40,960 for VCA). One of these serum samples was shown by immunoprecipitation to recognize at least 11 EBV-specific proteins from virus producer cells labeled in vivo and 10 EBV-specific proteins from in vitro translations of producer cell mRNA. Six of the in vivo labeled proteins (135,000, 89,000, 50,000 to 55,000 doublet, 46,000, and 34,000 daltons) are "early" by their resistance to phosphonoacetic acid, and five (350,000, 220,000, 160,000, 140,000, and 85,000 daltons) are "late" membrane and capsid proteins. The EBV-specific proteins immunoprecipitated from in vitro translations had molecular masses of 150,000, 140,000, 115,000, 52,000, 50,000, 45,000, 34,000, 29,000, 17,000, and 15,000. Subcellular fractionation studies of cells labeled in vivo revealed that the 135,000-dalton protein and part of the 50,000 to 55,000 dalton protein doublet were found in both the nuclear and the cytoplasmic fractions, and thus are good candidates to be components of the EA(D) diffuse-type immunofluorescence observed with many EA-positive sera. PMID- 6294184 TI - A sensitive method for detecting cells coated with antibodies with human monoclonal rheumatoid factor produced in vitro. AB - A human monoclonal rheumatoid factor (RF) produced in vitro by an Epstein-Barr virus immortalized cell line has been used to detect and quantitate antibodies specifically bound to cells. The RF is purified from cell culture supernatant by a simple procedure and then radiolabeled. In this assay, the binding of 125I labeled RF to cells coated with specific antibodies is determined. This RF method detects very low titers of antibodies directed against specific cellular antigens and also minorities of antibody coated tumor cells in mixtures with identical non coated cells. The advantages of this unique human monoclonal antibody are discussed. PMID- 6294183 TI - Autoradiographic detection of IgG and viral antigens. AB - Autoradiographic methods can be used as an alternative to indirect immunofluorescence to detect viral antigen expression or the presence of IgG in tissue sections. Iodinated protein A isolated from Staphylococcus aureus detects an influx of IgG into the central nervous system of mice inoculated with the coronavirus SD. Antispecies antibody that has been iodinated detects coronavirus antigen expression for 24 days post-inoculation while it is only detectable for 10 days by immunofluorescence. A direct comparison of indirect fluorescence and autoradiographic methods indicates that the autoradiographic techniques are considerably more sensitive. This increased sensitivity is sufficient to permit the detection of viral antigen in formalin fixed paraffin embedded tissue sections. PMID- 6294185 TI - Purification of human blood monocytes by hypotonic density gradient centrifugation in Percoll. AB - Leukocyte cell concentrates, obtained by continuous flow leukapheresis from single donors, were separated on a continuous hypotonic (260 mosM) Percoll gradient. On average, 86% of monocytes were recovered in a sharp band at a purity of up to 91% (average 76%). By this procedure 1-2 X 10(9) monocytes may be obtained from an individual donor. Hypotonic gradient purification, as compared with isotonic (295 mosM) conditions, proved superior with regard to capacity, speed of performance, yield and monocyte purity. PMID- 6294186 TI - [Diagnosis, prevalence, prognosis and treatment of the trophoblastic component in germinal tumours of the testis in the adult]. AB - The authors report a series of 97 germinal tumours of the testis in the adult, studied first in terms of conventional histological data. Of 33 seminomas, 2 secreted HCG. The 5-year actuarial survival at stage I was 93%, and at stage II 75%. Stage II deaths revealed the existence of not purely seminomatous tumours. Amongst dysembryomas, half secreted HCG, with 3 histological groups: predominant choriocarcinomas, tumours with a trophoblastic component and "apparently pure" dysembryomas. The 3-year actuarial survival for dysembryomas was 90% at stage I and 58% at stage II. 51 patients of the series were studied retrospectively by sections with HCG peroxidase, a technique which reveals the intracytoplasmic synthesis of the hormone. Two types of cells have proved capable of such synthesis: syncytial cells, of syncytial-trophoblastic type, and small mononuclear cells. One third of seminomas and 90% of dysembryomas proved to have a trophoblastic component as demonstrated by HCG immunoperoxidases. All patients secreting HCG were HCG peroxidase positive. This equally applied to all patients with syncytial cells. Furthermore, all the indications are that HCG secretion is above all by the syncytial cells. From a diagnostic standpoint, any rise in beta HCG is synonymous with an HCG immunoperoxidase trophoblastic component. Detection of such a component using immunoperoxidase would seem to be essential for non secreting tumours. From a prognostic standpoint, seminomas with a trophoblastic component are in fact dysembryomas and lymph node dissection should be performed, this being the only way of not missing a non-seminomatous metastasis. Therapeutically, pure seminomas are distinguished by the possibility of cure by radiotherapy. For all other tumours, orchidectomy must be followed by lymph node dissection. Subsequent treatment is decided on the basis of the results of the latter, with the exception of tumours with visceral metastases where chemotherapy must come first. PMID- 6294187 TI - [Primary vesical adenocarcinoma with signet ring cells or linitis plastica of the bladder. Three cases]. AB - Signet ring cell adenocarcinomas of the bladder are regarded as rare tumours. The authors actually think that their frequency is underrated since they report three new cases out of a range of 700 bladder tumours. Revealing signs consist in urgency pain, frequency and/or hematuria. This set of cancer is usually tardily diagnosed and therefore of very poor prognosis whatever the therapies. PMID- 6294188 TI - [Nephroblastoma in the adult. Apropos of a case]. AB - Nephroblastoma is a rare tumour in the adult and the diagnosis is only rarely made preoperatively. In the light of a new case, the authors consider the problems of histopathological differential diagnosis with other borderline forms. They study the clinical aspects of these tumours. The prognosis would seem to be extremely gloomy, worse than that of renal adenocarcinoma. PMID- 6294190 TI - Virus-like intranuclear particles in bronchiolar-alveolar cell carcinoma. PMID- 6294191 TI - [Biosynthesis and intracellular transport of virus membrane glycoproteins]. PMID- 6294189 TI - Neuropeptides in the pathogenesis of obesity-associated benign acanthosis nigricans. AB - The association of acanthosis nigricans with pituitary tumors and insulin resistant diabetes suggests that a pituitary peptide may promote papillomatosis and acanthosis characteristic of acanthosis nigricans. Although such a peptide has not been isolated, it may derive by sequential cleavage from the 31,000 dalton precursor peptide to ACTH and beta-lipotropin (beta-LPH). In order to evaluate the role of pituitary peptides in the pathogenesis of acanthosis nigricans, we compared plasma levels of beta-endorphin (beta-EP) and ACTH in plasma of 8 fasting patients with obesity-associated benign acanthosis nigricans and 7 fasting normal controls utilizing sensitive radioimmunoassay procedures. Mean plasma beta-EP levels for the acanthosis nigricans and control subjects were not significantly different (90 pg/ml vs. 140 pg/ml), nor was any significant difference observed between plasma ACTH levels of the 2 groups (42.3 and 31.2 pg/ml, respectively.) Our data indicate that plasma levels of the pituitary derived peptides ACTH and beta-EP are not increased in obesity-associated benign acanthosis nigricans, and suggest that its proposed hormonal mediator might originate independently from the large peptide precursor of ACTH, beta-LPH and their fragments. PMID- 6294192 TI - [Studies on changes of the plasma prolactin, growth hormone and ACTH levels following surgical stress and epidural micro-injections of morphine hydrochloride as a postoperative analgesic method]. AB - Epidural micro-injection of morphine hydrochloride is very useful for the postoperative analgesic method. In this study, we investigated the relationship between appearance of amelioration of pain and the plasma prolactin (PRL), growth hormone (GH) and ACTH levels following epidural morphine. The plasma PRL levels significantly elevated with surgical stress. Following epidural morphine, the plasma PRL levels significantly decreased in the effective cases, and significantly increased in the ineffective cases. Following epidural morphine, the plasma ACTH levels significantly decreased in the effective cases, and insignificantly increased in the ineffective cases. The duration to onset of amelioration of pain was prolonged with intravenous injections of metoclopramide (MCP) before epidural morphine. We concluded that patterns of PRL and ACTH releases following epidural morphine were correspondent with the analgesic effect, and that appearance of amelioration of pain following epidural morphine may depend upon the dopaminergic mechanism. PMID- 6294193 TI - [Studies on the recurrent trophoblastic disease]. AB - We studied the remission and the recurrent of trophoblastic disease treated in our clinic, during 1973 to 1980. 1) The remission and recurrent rates of 168 cases were 91.1% (153/168) and 5.2% (8/153) respectively. In these percentages, high risk was 65.9% (29/44) and 20.7% (6/29), low risk was 100% (124/124) and 1.6% (2/124), choriocarcinoma was 58.3% (21/36) and 28.6% (6/21), invasive mole was 100% (44/44) and 0, and undetermined was 100% (88/88) and 2.3% (2/88). 2) The interval to recurrence: 5 cases appeared under a year, 3 cases did over a year. And remission rate of recurrent case was (4/8), high risk was (2/6), and low risk was (2/2). 3) HCG-beta titers when U-hCG decreased under the LH levels was higher until 3 months in the recurrent case of high risk than that unrecurrent, while we did not recognize any differences after 3 months. Therefore criteria of remission in high risk would occurred on continuous LH levels of U-hCG for 3 months. 4) High risk has been treated by 3 courses of additional chemotherapy, but that when hCG-beta titer over 1.0ng/ml require more chemotherapy. PMID- 6294194 TI - [Studies on luteinizing hormone receptors of human corpora lutea during menstrual cycle and pregnancy]. AB - With the purpose of explicating the lifespan of human corpora lutea, using human corpora lutea of the menstrual cycle and pregnancy, binding of 125I-LH to the 20,000g cell membrane fraction was examined. 1) Specific bindings of 125I-LH, 125I-HCG were demonstrated in the 20,000g cell membrane fraction. Although LH and HCG were parallel in inhibiting 125I-LH binding, HCG was found to be more effective. FSH did not inhibit binding. 2) Binding of 125I-LH was dependent on time, temperature, 125I-LH concentration, amount of the cell membrane fraction protein and pH. The highest binding was seen at pH 6.0 while incubating for 60 min at 37 degrees C. 3) The number of LH receptors in human corpora lutea of the menstrual cycle increased towards midluteal phase, especially on 5th day from ovulation, and decreased towards late luteal phase. LH receptor was not found in corpus albicans. The apparent dissociation constant of each corpus luteum did not change throughout the menstrual cycle. 4) Corpora lutea of pregnancy contained a few or no receptors which bound 125I-LH specifically. These data suggest that LH receptor is an important factor regulating the lifespan of corpus luteum and exogenous HCG has effect on luteal insufficiency, but the effect of HCG on threatened abortion is uncertain. PMID- 6294195 TI - [Definition, classification and diagnostic standards for trophoblastic diseases]. PMID- 6294196 TI - [Clinical diagnosis of well differentiated cervical adenocarcinoma with plentiful mucous secretion]. AB - Six cases of cervical adenocarcinoma with plentiful mucous secretion were recently experienced. They were clearly distinguishable from the majority of adenocarcinoma without plentiful mucous secretion from the standpoint of clinical and colposcopic findings. As common features deduced from clinical and colposcopic findings, it could be pointed out that portio vaginalis become swollen as a whole showed resiliency as of a rubber ball, and that erosion widespread over nearly entire surface of portio vaginalis is accompanied with a plentiful mucus. On the surface of erosion was found neither localization of ulcer, nor necrosis and nor bleeding trend even in cases under progressing. Colposcopic findings were distinguished with features of papillea-, net-and transformation zone-like, and mixed type. On the other hand, all the cases were evaluated cytologically to be positive with difficulty. And they were histologically well differentiated adenocarcinoma with plentiful mucus. The distinctive feature in the clinical and colposcopic findings identified for the cases of well differentiated adenocarcinoma with a plentiful mucus was evaluated to be useful means to assist the diagnosis of cervical adenocarcinoma. PMID- 6294197 TI - [Gonadotropin and plasma cyclic nucleotides]. AB - Plasma cyclic nucleotides in some physiological conditions were measured in normal women and the changes of cyclic AMP were studied after the loading of gonadotropins in normal and castrated women. We demonstrated a circadian rhythm of cyclic AMP and its variation during menstrual cycle in normal women and also demonstrated its significant increase in pregnant women. But there was no significant variation of plasma cyclic GMP levels in those physiological conditions. Plasma cyclic AMP was increased by the simultaneous administration of HMG and HCG in both normal and castrated women. However, individual administration of HMG or HCG demonstrated that it was HCG that caused that increase in plasma cyclic AMP even in castrated women. To examine the extragonadal effects of HCG, cortisol, triiodothyronine (T3), thyroxine (T4) in the plasma were measured after the loading of HCG. The plasma cortisol level was influenced by HCG in neither normal nor castrated women. HCG did not cause any change of T3 or T4 in normal women; however it caused a marked increase in T3 with a small decrease in T4 in castrated women. PMID- 6294198 TI - [Diagnosis of malignant fibrous histiocytoma]. PMID- 6294199 TI - [A case of glucagonoma]. PMID- 6294201 TI - Na,K-ATPase inhibitor from guinea pig brain is not ouabain-like. AB - An acetone HCl extract of guinea pig brain reported to have ouabain-like activity was prepared. The extract was column-chromatographed on Sephadex G-10 and fractions were tested for ion content and "apparent ouabain-like" activity, i.e., ability to displace ouabain from its binding site and to inhibit Na,K-ATPase under conditions that optimize one or the other assay. "Apparent ouabain-like" activity eluted with the salt peak. When the ability of these fractions to inhibit ouabain binding was correlated with their potassium concentrations, the displacement could be entirely accounted for by the potassium content. The Na,K ATPase inhibiting activity of these salt peak fractions was demonstrable in a Tris assay medium but not in a histidine (metal chelator)assay medium. Thorough desalting of fractions with "apparent ouabain-like" activity abolished this activity. Additional Na,K-ATPase inhibitory activity appeared in fractions distinct from the salt peak. However, these fractions did not display ouabain displacing ability. It is concluded that the "apparent ouabain-like" activity of the guinea pig brain extract is not truly ouabain-like and can be explained by interfering ions. PMID- 6294200 TI - [A case of myxedema associated with cerebellar ataxia and various neurological findings]. PMID- 6294202 TI - Measurement of human converting enzyme level by direct radioimmunoassay. AB - CE was purified from human lung, and antisera were raised in rabbits. Antisera inhibited the activity of the purified enzyme from lung and kidney and the plasma CE of normal persons and sarcoid patients. With antisera at a titer of 1:100,000, a sensitive, direct RIA was developed. CE purified from lung or kidney and CE present in normal and in sarcoid plasma gave parallel logit-log displacement lines, suggesting immunological identity. The level of CE in normal human plasma was 400 +/- 131 ng/ml. In untreated sarcoid patients, the enzyme level and activity increased in parallel. There was a negative correlation (r = -0.81) between enzyme level and diffusing capacity of the lung for CO in sarcoid patients. Synthetic inhibitors such as captopril or MK 421 did not interfere with the RIA, permitting enzyme levels to be monitored in patients undergoing acute inhibitor therapy. During administration of MK 421, CE activity was negligible and plasma levels of CE did not change. In contrast, renin activity increased eightfold during the inhibitor therapy. PMID- 6294203 TI - Glucose: a role as a free radical scavenger in biological systems. AB - Recent observations indicate that OH . may be important in the microbicidal capacity of phagocytic cells, in prostaglandin metabolism, and as a mediator of inflammation. Although glucose is a weak hydroxyl scavenger, it occurs in high concentrations in biological systems. We therefore studied the capacity of glucose to scavenge OH . in biological systems known to generate this reactive oxygen species. Our experiments used a specific assay for the detection of OH .. We measured 14CO2 released during the oxidation of 14C-benzoic acid. We have previously demonstrated that benzoic acid is oxidized as a consequence of OH . in the following systems: the enzyme system xanthine-xanthine oxidase, zymosan stimulated granulocytes, and arachidonic acid-stimulated platelets as a consequence of the lipoxygenase pathway. In all three systems the oxidation of benzoic acid was inversely proportional to the concentration of glucose in the assays. Also, platelets incubated with arachidonic acid and a high concentration of glucose increased HETE production, an effect predicted by the capacity of glucose to act as an OH . scavenger. Our results indicate that glucose acts as a scavenger of OH . in physiological concentrations and therefore may serve an antioxidant role in biological systems. In addition, the capacity of glucose to act as an OH . scavenger may explain some of the defects seen in patients with diabetes mellitus. PMID- 6294204 TI - Pathology as it related to ear surgery. I. Surgery of glomus tumours. PMID- 6294205 TI - Pleomorphic adenoma of the auricle. PMID- 6294206 TI - Granular cell myoblastoma of the larynx. PMID- 6294207 TI - Chemodectoma of the larynx. PMID- 6294208 TI - Characteristics of thyrotrophin-stimulated cyclic AMP production in cultured human thyroid cells from thyrotoxic tissue and non-toxic goitres. AB - We have described a system for the maintenance in culture of isolated human thyroid cells from both thyrotoxic tissue and non-toxic goitres. The cells isolated from the two thyroid tissue types showed similar cyclic AMP response characteristics to TSH with large increases in intracellular and extracellular cyclic AMP after 20-min incubations. Maximal responses were obtained with 50 mu. TSH/ml and half-maximal responses at 1.0 mu. TSH/ml. With cell passage the cyclic AMP responses to TSH decreased in magnitude and sensitivity. As with other thyroid cultures, growth of the cells with TSH induced arrangement into follicular structures, whereas cells grown in the absence of TSH remained as a monolayer. Basal intracellular cyclic AMP levels were increased in a dose-related fashion in cells grown in the presence of graded concentrations of TSH and the maximal response to further additions of TSH was not greater than in control cultures. PMID- 6294209 TI - Hyperprolactinaemia attenuates the gonadotrophin releasing hormone receptor response to gonadectomy in rats. AB - Measurement of pituitary gonadotrophin releasing hormone (Gn-RH) receptor content provides a qualitative index of prior exposure of the pituitary gland to endogenous Gn-RH. The effect of moderate hyperprolactinaemia (serum prolactin = 95-250 micrograms/1), achieved with three pituitary grafts beneath the renal capsule, on the pituitary Gn-RH receptor content and serum LH responses to gonadectomy of adult rats has been studied. In males the presence of hyperprolactinaemia for 7 days completely prevented the increase in Gn-RH receptor content 3 days after castration and inhibited the serum LH rise by 45%. By 6 days after castration, Gn-RH receptors had increased in the hyperprolactinaemic castrated animals but values were 33% lower than in sham grafted controls, while the serum LH increase attenuated by 30%. Pituitary LH content was also lower in grafted castrated animals 6 days after castration. Hyperprolactinaemia for 3 weeks had no effect on Gn-RH receptors or pituitary LH content of intact male rats, although basal serum LH was decreased by 50%. Hyperprolactinaemia also attenuated the increases in Gn-RH receptors, serum LH and pituitary LH which occurred 6 days after ovariectomy in female rats. In all experiments the pituitary content of prolactin was reduced by 80-90% in animals bearing pituitary grafts. These results suggest that hyperprolactinaemia restricts the Gn-RH receptor response to gonadectomy by decreasing endogenous hypothalamic Gn-RH secretion. PMID- 6294210 TI - Central nervous integration of cardiovascular control. PMID- 6294211 TI - Characterization of a membrane pore-forming protein from Entamoeba histolytica. AB - We describe the partial purification and characterization of a pore-forming material (PEM) from Entamoeba histolytica. The formation of ion channels by PFM was examined in three systems. (a) PFM depolarizes J774 macrophages and mouse spleen lymphocytes as measured by [3H]TPP+ uptake. (b) PFM induces rapid monovalent cation flux across the membrane of phosphatidylcholine-cholesterol vesicles. (c) PFM confers a voltage-dependent conductance to artificial planar bilayers, which is resolved as a summation of opening of individually conducting steps of 67 pS in 0.1 M KCl. Monomers of PFM are functional; however, a preferential aggregation occurs in the planar bilayer. Activity is pronase, trypsin, and heat sensitive and is stable between pH 5-8. PFM is not secreted by unstimulated amoebae but after exposure to the calcium ionophore A23187, concanavalin A, and E. coli lipopolysaccharide, 5-10% of the total cell content of PFM is released into the medium within 5-10 min. High-performance gel filtration results in an approximately 1,000-fold purification of PFM and gives an Mr of 30,000. This protein may play a role in the cytotoxicity mediated by E. histolytica. PMID- 6294212 TI - Limiting dilution analysis of Epstein-Barr virus-induced immunoglobulin production by human B cells. AB - The Epstein-Barr virus (EBV) is a herpes virus that has the capacity to infect human B cells and to induce them to secrete immunoglobulin (Ig). In the current experiments, Poisson analysis of limiting dilution cultures has been used to study the activation of human peripheral B cells by the B95-8 strain of EBV. Under the culture conditions used, 0.2-1% of peripheral blood B cells were activated by EBV to secrete IgM or IgG. In addition, when multiple replicate cultures containing limited numbers of B cells were tested for IgM and for IgG production, the precursors for IgM and IgG segregated independently; thus, individual B cell precursors matured into cells secreting IgM or IgG but not both classes of Ig. Additional experiments using limiting dilutions of EBV were undertaken to study the viral requirements for B cell activation. These studies indicated that B cell activation by EBV to produce Ig was consistent with a "one hit" model and inconsistent with a "two-hit" model. Taken together, these results indicate that infection by one EBV virion is sufficient to induce a precursor peripheral blood B cell to secrete Ig and that only one isotype of Ig is then secreted. PMID- 6294214 TI - An assessment of the immunofluorescence technique as a method for demonstrating the histological localization of tetrahydrocannabinol in mammalian tissues. AB - The use of the indirect immunofluorescence technique as a method for demonstrating the histological localization of tetrahydrocannabinol (delta-THC) has been examined. The experimental protocol was designed in order that optimal staining conditions with respect to temperature, the length of time of incubations and washes, and the dilution of the antisera should be defined. No marked differences were detected between frozen sections of liver from normal and delta-THC-injected mice. Results from radiotracer experiments using human liver suggest that the success of the method is dependent upon the solubility characteristics of the antigen-antibody complex. PMID- 6294213 TI - Analysis of the defects responsible for the impaired regulation of Epstein-Barr virus-induced B cell proliferation by rheumatoid arthritis lymphocytes. I. Diminished gamma interferon production in response to autologous stimulation. AB - T cells of patients with rheumatoid arthritis (RA) do not control the rate of B lymphoblast transformation induced by Epstein-Barr virus (EBV) as efficiently as T cells from healthy individuals; thus, lymphoblast cell lines are established more readily in RA lymphocytes in vitro after EBV infection. In the present experiments, we have asked whether this T cell regulation can be reproduced by lymphocytes. We found that normal T cells, activated in allogeneic or autologous mixed leukocyte reactions (MLR), produce lymphokines that inhibit in vitro EBV induced B cell proliferation. Allogeneic MLR supernatants inhibited EBV-induced DNA synthesis 62 +/- 4% (mean +/- SE) at 10 d post-infection, whereas autologous MLR supernatants suppressed it 50 +/- 3%. RA T cell supernatants produced in an allogeneic MLR suppressed as well as normal T cell supernatants (64 +/- 5% inhibition). In contrast, supernatants from RA autologous MLR had little inhibitory activity. EBV-induced DNA synthesis at 10 d was reduced only 8 +/- 3%, compared with the 50 +/- 3% suppressive activity of normal autologous MLR supernatants. The magnitude of the proliferative responses in the autologous MLR regenerating the lymphokines was similar in the normal and RA populations. After depletion of adherent cells from the RA auto-MLR stimulators, supernatant inhibitory activities increased to normal levels (from 11 +/- 6 [SE] to 52 +/- 6% [SE]). The inhibitory factor involved in the regulation of in vitro EBV infection is a protein with a molecular weight of approximately 50,000. Its activity is eliminated by hearing at 56 degrees C and by exposure to acid at pH 2. The inhibitory activity is blocked by mixing the MLR supernatants with a polyvalent antisera or monoclonal antibodies specific for human gamma interferon. Gamma interferon produced by activating T cells in allo- or auto-MLR can reproduce T cell-mediated regulation of EBV-induced B cell proliferation, and the failure of RA auto-MLR to generate that lymphokine parallels the defective T cell regulation of EBV-induced B cell proliferation characteristic of RA lymphoid cells. PMID- 6294215 TI - Incidence and toxicological aspects of drugs detected in 484 fatally injured drivers and pedestrians in Ontario. AB - Results are presented of a comprehensive drug study carried out on specimens from drivers and pedestrians fatally injured in Ontario. Toxicological analyses were regularly performed on blood and urine and occasionally on vitreous humor, stomach contents, and liver. The analytical procedures could detect and quantitate a wide variety of drugs including such illicit drugs as Cannabis. With respect to drivers, alcohol was found in 57% of the study sample and drugs other than alcohol, in 26%. However, in only 9.5% of the drivers were psychoactive drugs (other than alcohol) detected in the blood in concentrations that may adversely affect driving skills. Delta-9-Tetrahydrocannabinol and diazepam accounted for a majority of the findings in this category. PMID- 6294217 TI - [Histocompatibility profile of hepatoma in Taiwan]. PMID- 6294216 TI - [Seroepidemiology of hepatitis A infection in children in Taiwan]. PMID- 6294218 TI - [Metastatic adenocarcinoma to the lungs simulating diffuse type bronchiolo alveolar carcinoma--a case report]. PMID- 6294219 TI - Simulation of Na channel inactivation by thiazine dyes. AB - Some dyes of the methylene blue family serve as artificial inactivators of the sodium channels when present inside squid axons at a concentration of approximately 0.1 mM. The dyes restore a semblance of inactivation after normal inactivation has been destroyed by pronase. In fibers that inactivate normally, the dyes hasten the decay of sodium current. Many dye-blocked channels conduct transiently on exit of the dye molecule after repolarization to the holding potential. In contrast, normally inactivated channels do not conduct during recovery from inactivation. Kinetic evidence shows that inactivation of a dye blocked channel is unlikely or impossible, which suggests that dye molecules compete with inactivation "particles" for the same site. In the absence of tetrodotoxin, the dyes do not affect the ON gating current unless the interpulse interval is very short. If sufficient equilibration time is allowed during a pulse, the initial amplitude of the OFF gating current is reduced to near zero. This suggests that a dye molecule is a Na channel completely blocks that channel's gating current, even the fraction that is resistant to normal inactivation. Dyes block INa and Ig with the same time course. This provides the strongest evidence to date that virtually all of recorded "gating current" is associated with Na channels. Tetrodotoxin greatly slows dissociation of dye molecules from Na channels and reduced gating current during both opening and closing of the channels. PMID- 6294220 TI - Block of squid axon K channels by internally and externally applied barium ions. AB - We have studied the interactions of Ba ion with K channels. Ba2+ blocks these channels when applied either internally or externally in millimolar concentrations. Periodic depolarizations enhance block with internal Ba2+, but diminish the block caused by external Ba2+. At rest, dissociation of Ba2+ from blocked channels is very slow, as ascertained by infrequent test pulses applied after washing Ba2+ form either inside or outside. The time constant for recovery from internal and external Ba2+ is the same. Frequent pulsing greatly shortens recovery time constant after washing away both Ba2+in and Ba2+out. Block by Ba2+ applied internally or externally is voltage dependent. Internal Ba2+ block behaves like a one-step reaction governed by a dissociation constant (Kd) that decreases e-fold/12 mV increase of pulse voltage: block deepens with more positive pulse voltage. For external Ba2+, Kd decreases e-fold/18 mV as holding potential is made more negative: block deepens with increasing negativity. Millimolar external concentrations of some cations can either lessen (K+) or enhance (NH+4, Cs+) block by external Ba2+. NH+4 apparently enhances block by slowing exist of Ba ions from the channels. Rb+ and Cs+ also slow clearing of Ba ions from channels. We think that (a) internally applied Ba2+ moves all the way through the channels, entering only when activation gates are open; (b) externally applied Ba2+ moves two-thirds of the way in, entering predominantly when activation gates are closed; (c) at a given voltage, Ba2+ occupies the same position in the channels whether it entered from inside or outside. PMID- 6294221 TI - Modified kinetics and selectivity of sodium channels in frog skeletal muscle fibers treated with aconitine. AB - The effect of the plant alkaloid aconitine on sodium channel kinetics, ionic selectivity, and blockage by protons and tetrodotoxin (TTX) has been studied in frog skeletal muscle. Treatment with 0.25 or 0.3 mM aconitine alters sodium channels so that the threshold of activation is shifted 40-50 mV in the hyperpolarized direction. In contrast to previous results in frog nerve, inactivation is complete for depolarizations beyond about -60 mV. After aconitine treatment, the steady state level of inactivation is shifted approximately 20 mV in the hyperpolarizing direction. Concomitant with changes in channel kinetics, the relative permeability of the sodium channel to NH4,K, and Cs is increased. This altered selectivity is not accompanied by altered block by protons or TTX. The results suggest that sites other than those involved in channel block by protons and TTX are important in determining sodium channel selectivity. PMID- 6294222 TI - Biphasic regulation of development of the high-affinity saxitoxin receptor by innervation in rat skeletal muscle. AB - Specific binding of 3H-saxitoxin (STX) was used to quantitate the density of voltage-sensitive sodium channels in developing rat skeletal muscle. In adult triceps surae, a single class of sites with a KD = 2.9 nM and a density of 21 fmol/mg wet wt was detected. The density of these high-affinity sites increased from 2.0 fmol/mg wet wt to the adult value in linear fashion during days 2-25 after birth. Denervation of the triceps surae at day 11 or 17 reduced final saxitoxin receptor site density to 10.4 or 9.2 fmol/mg wet wt, respectively, without changing KD. Denervation of the triceps surae at day 5 did not alter the subsequent development of saxitoxin receptor sites during days 5-9 and accelerated the increase of saxitoxin receptor sites during days 9-13. After day 13, saxitoxin receptor development abruptly ceased and the density of saxitoxin receptor sites declined to 11 fmol/wg wet wt. These results show that the regulation of high-affinity saxitoxin receptor site density by innervation is biphasic. During the first phase, which is independent of continuing innervation, the saxitoxin receptor density increases to 47-57% of the adult level. After day 11, the second phase of development, which is dependent on continuing innervation, gives rise to the adult density of saxitoxin receptors. PMID- 6294223 TI - Effect of Ca2+, cyclic GMP, and cyclic AMP added to artificial solution perfusing lingual artery on frog gustatory nerve responses. AB - The lingual artery of the bullfrog was perfused with artificial solution and the effects of Ca2+, Ca-channel blockers (MnCl2 and verapamil), cGMP, and cAMP added to the perfusing solution of the gustatory nerve responses were examined. The responses to chemical stimuli of group 1 (CaCl2, NaCl, distilled water, D galactose, and L-threonine) applied to the tongue surface were greatly decreased by a decrease in Ca2+ concentration in the perfusing solution, suppressed by the Ca-channel blockers, enhanced by cGMP, and suppressed by cAMP. The responses to chemical stimuli of group 2 (quinine hydrochloride, theophylline, ethanol, and HCl) were practically not affected by a decrease in Ca2+ concentration, the Ca channel blockers, cGMP, and cAMP. The responses to the stimuli of group 1 seem to be induced by Ca influx into a taste cell that is triggered by depolarization and modulated by the cyclic nucleotides in a taste cell. The responses to group 2 seem to be induced without accompanying Ca influx. PMID- 6294224 TI - Sodium pump-mediated ATP:ADP exchange. The sided effects of sodium and potassium ions. AB - Resealed human red cell ghosts containing caged ATP (Kaplan et al., 1978) and [3H]ADP were irradiated at 340 nm. The photochemical release of free ATP initiated a rapid transphosphorylation reaction (ATP:ADP exchange), a component of which is inhibited by ouabain. The reaction rate was measured by following the rate of appearance of [3H]ATP. The sodium pump-mediated ATP:ADP exchange reaction showed high-affinity stimulation by Mg ions (less than 10 microM) and was inhibited at higher levels. At optimal [Mg], extracellular Na (Nao) had a biphasic effect. Nao progressively inhibited the reaction rate between 0 and 10 mM and stimulated at higher levels. Intracellular Na (Nai) activated the reaction; the rate was maximal when Nai was 1 mM and remained unaltered up to 115 mM Nai at constant Nao. Extracellular K ions (Ko) inhibited the reaction; at high Nao, half-maximal inhibition was observed with 0.9 mM Ko. Lio inhibited the exchange rate with a lower affinity than Ko; half-maximal inhibition was produced by approximately 50 mM Lio. Intracellular K ions were without dramatic effect on the reaction rate in the concentration range where Ko inhibited completely. The relationship between these observations and previous studies on porous preparations is discussed, as well as the extent to which these observations support the hypothesis that the sodium pump-mediated ATP:ADP exchange reaction accompanies the Na:Na exchange transport mode of the sodium pump. PMID- 6294225 TI - Transmembrane respiration-driven H+ translocation is unimpaired in an eup mutant of Escherichia coli. PMID- 6294226 TI - Neuraminidase-sensitive erythrocyte receptor for enterovirus type 70. AB - Enterovirus type 70 (EV70) agglutinated human 'O' erythrocytes at 4 degrees C as well as 22 degrees C, but visible agglutination was lost when warmed at 37 degrees C although the virus remained attached to the surface of the erythrocyte. The receptor sites for the virus were neuraminidase-sensitive. A direct involvement of sialic acid on the cell surface in virus-cell interaction was confirmed by the fact that the presence of fetuin or free N-acetylneuraminic acid inhibited the haemagglutinating activity of EV70. Similar numbers of virus particles were required for 1 haemagglutinating unit (HAU) of EV70 and 1 HAU of mengovirus, whereas 2.6-fold or more of virus particles of echovirus type 7 and type 11 gave the same activity. On the other hand, the number of receptor sites on the cell surface for EV70 was found to be sevenfold more than for mengovirus. Therefore, the erythrocyte receptor for EV70 is different from that for common enteroviruses and similar, though not identical, to the cardiovirus receptor. However, serological tests such as neutralization, complement fixation or haemagglutination inhibition did not reveal any common antigen between EV70 and cardiovirus. PMID- 6294228 TI - Restricted translation of the genome of the flavivirus Kunjin in vitro. AB - Virion RNA of Kunjin virus was translated in rabbit reticulocyte lysates at a rate (7000 daltons/min) approaching that observed previously in vivo. As many as 18 polypeptides were translated and shown by tryptic peptide mapping to be closely related to one another and to contain some of the elements of envelope (E) and most of the elements of core (C) proteins of Kunjin virus. None of the products resolved in gels were precipitated by antiserum to purified E protein, but the larger products were precipitated by an antiserum against all the Kunjin virus-specified proteins. No evidence was obtained of translation of the non structural proteins, despite attempts to retain or perturb the structure of the virion RNA. As many as 50% of the amino acid sequences in the peptide maps were not identifiable. PMID- 6294227 TI - Influence of the cytoskeleton on the expression of a mouse hepatitis virus (MHV 3) in peritoneal macrophages: acute and persistent infection. AB - The effects of anti-cytokinetic drugs on virus production, formation of syncytia, cell surface changes and lysosomal damage were examined during mouse hepatitis virus 3 (MHV-3) infection of mouse peritoneal macrophages. Colchicine and vinblastine caused no detectable effect on the infectious process. In the presence of cytochalasin B the acute, highly cytopathogenic interaction that MHV 3 establishes with macrophages was converted into one which persisted for several days. Under these conditions the cell surface changes induced by the infection were maintained unaltered but cell fusion was reduced and no significant lysosomal damage was detectable. PMID- 6294229 TI - Development of an enzyme-linked immunosorbent assay for the identification of arthropod-borne togavirus antibodies. AB - The applicability of the standard enzyme-linked immunosorbent assay (ELISA) for the identification of togavirus infections was investigated. Optimal concentration of gradient-purified antigen was 2.5 micrograms/well for alphaviruses or flaviviruses when coating polystyrene microtitre plates. A procedure for producing antigen in suckling mouse brain was developed. Results obtained with ELISA could be correlated with standard serology, but in general the ELISA was more sensitive. The ELISA was specific in differentiating alphavirus antigens. Flavivirus cross-reactivity was magnified by ELISA. ELISA should be useful as a rapid screening assay for non-related antigens, avoiding the extensive techniques currently used in standard serodiagnosis. PMID- 6294230 TI - Molecular cloning of the endogenous rat C-type helper virus DNA sequence: structural organization and functional analysis of some restricted DNA fragments. AB - Recently, we have identified and purified the integrated and proviral DNA sequences specific for two endogenous rat type C leukaemia helper viruses: WR RaLV which originated from a fibrosarcoma induced in a feral rat and RHHV from the cell line HTC-H1 which originated from a Buffalo rat hepatoma. The rat leukaemia helper virus DNA sequences have previously been shown to be 8.4 to 8.8 kilobases (kb) in size. In this communication, we report the molecular cloning of the 8.8 kb DNA of RHHV by ligation at the BamHI site of the vector pBR322, cultured in an Escherichia coli RR1 host. After screening 5750 clones for ampicillin resistance and tetracycline sensitivity and testing by colony hybridization using 32P-labelled RHHV cDNA, four clones were isolated, two of which carried the total 8.8 kb DNA. A detailed restriction endonuclease map of the cloned RHHV DNA was deduced by sequential digestions of either 3'- or 5' labelled DNA. Of the 14 restriction enzymes tested, EcoRI, BamHI, PstI, KpnI, TaqI, PvuII and SmaI gave informative cleavage patterns. At least two copies of long terminal repeated sequences (LTR) flanking the 3' and 5' termini of the proviral DNA were identified by TaqI and PstI cleavages. LTR in the rat endogenous leukaemia helper virus DNA measured 780 +/- 20 nucleotides in length. The genetic information encoded by the cloned DNA was also analysed by hybridization selection of RHHV mRNA, which was then used in cell-free protein synthesis in a rabbit reticulocyte lysate system. Essentially all major RaLV specific proteins precipitable by anti-RaLV serum were synthesized in vitro, confirming that the RHHV genomic DNA was successfully cloned with little fidelity loss or scrambling of the genetic information. PMID- 6294231 TI - Ultraviolet irradiation of murine cytomegalovirus. AB - Ultraviolet irradiation of murine cytomegalovirus (MCMV) caused a rapid dose related decline in virus infectivity, manifested by virus antigen induction, and in virus production as measured by plaque formation and infectious centre assay. The virus survival curve was multi-component, suggesting host cell-assisted reactivation. Multiplicity reactivation and photoreactivation of MCMV were not observed in these experiments. Productive infection was more sensitive to u.v. irradiation than was virus antigen production, indicating differential inactivation of virus functions. The effects of u.v. irradiation were similar in most respects to those reported for human cytomegalovirus. PMID- 6294232 TI - Biological activity of cloned rat endogenous C-type virus DNA transferred by microinjection. AB - The biological activity of a molecularly cloned DNA of a rat endogenous C-type leukaemia helper virus, RHHV, was assessed by intranuclear microinjection into normal rat kidney cells (NRK153). Release of rat C-type leukaemia helper viruses by the microinjected cells was examined by superinfection of Kirsten-transformed non-producer cells (K-NRK). Immediate release of helper leukaemia viruses at a very low level was observed only in the NRK153 m3 . 5/cir cells microinjected with the supercoiled form of RHHV DNA in toto, suggesting that the circular form of the virus DNA might have expedited the replication and expression of virus particles. Genome rescue experiments were also performed by co-cultivating the microinjected NRK153 m cells carrying various linear RHHV DNAs, in toto or of subgenomic sizes, with K-NRK cells. The results indicated that both the total and the 5.8 to 6.2 kb DNA fragment proximal to the 5' terminus of the cloned RHHV 8.8 kb DNA were able to rescue successfully a transforming replication-competent pseudotype virus. Subgenomic DNA fragments derived from the centre or the 3' end of the RHHV DNA were ineffective in the genome rescue experiments. PMID- 6294233 TI - Rauscher spleen focus-forming virus: biological properties and relationship to helper viruses. AB - A Rauscher virus (RV)-transformed erythroid cell line, RA-1, was shown to be a non-producer cell line. RA-1 cells express not only gp51-54 env-related glycoprotein, but also gp70, which is more closely related to gp51-54 coded by a recombinant env gene than to the MuLV-R gp70. RA-1 cells could be infected by Friend, Moloney and Gross viruses, but not by the homologous Rauscher murine leukaemia virus. Rescue of spleen focus-forming activity was obtained on infection of these cells with MuLV-F or MuLV-Mol, but not with MuLV-Gross. The RNA of the RV complex resembles closely that of Friend virus (FV). It contains a 32S, presumably defective, genome, which most likely is responsible for spleen focus formation, and a 35S helper virus genome. Oligonucleotide fingerprint data suggest that RV has evolved independently of FV. Erythroid early BFU-E cells of mice infected with RV of Friend helper virus-infected RA-1 cells were shown to require no addition of conditioned medium to form large erythroid colonies (BFU E) in the presence of only small amounts of erythropoietin. PMID- 6294234 TI - Host antigens on avian oncoviruses: presence on viruses produced by quail, duck and rat cells of antigens related to membrane antigens of chick embryo fibroblasts and chicken erythrocytes. AB - Avian sarcoma viruses (ASV) produced by Japanese quail embryo fibroblasts (QEF) were inactivated to the same degree as ASV produced by chick embryo fibroblasts (CEF), with or without complement, by rabbit antisera to CEF and to membrane antigens of chick embryo and adult chicken erythrocytes, in particular their unique age-specific antigens. ASV produced by duck embryo fibroblasts (DEF) were only inactivated by the antisera to the chicken erythrocyte antigens. Hence, viruses produced by QEF and DEF bear on their envelope antigens related to the chicken antigens picked up by ASV in CEF. These antigens are presumably coded by the quail and duck cells, since antigens related to the chicken antigens were found on QEF and on quail and duck erythrocytes. Antigens related to the chicken antigens have also been found on ASV shed in low amounts by semi-permissive rat sarcoma cells (line 17RBI77) and on the surface of these cells, as well as on that of another semi-permissive cell line (RBH) originating from a hamster sarcoma produced by inoculation of the rat cells. The origin of these antigens, which may play a role in semi-permissiveness, remains to be explained since they do not appear to be normally expressed by rat or hamster cells. It was also found that, contrary to an earlier conclusion and in agreement with a recent report, uninfected CEF bear on their membrane an antigen that is related or identical to the specific antigen of chick embryo erythrocytes. Therefore, only the antigen related to the adult-specific chicken erythrocyte antigen does not pre-exist on CEF. PMID- 6294236 TI - Spread of virus and distribution of latent infection following ocular herpes simplex in the non-immune and immune mouse. AB - In both non-immune and immune mice infected with herpes simplex virus the incidence of latent infection of the trigeminal ganglion was related to the severity of ocular virus infection. During primary infection, virus was shown to travel via the ophthalmic part of the ganglion to reach the brainstem, from where centrifugal spread resulted in latent infection of neurones in the trigeminal ganglion which did not serve the site of inoculation. Primary infection also resulted in latent infection of the superior cervical ganglion. Shedding of virus occurred rarely in the tears of animals which had recovered from primary disease. In immune mice, spread of virus resulted in a much lower incidence of latent infection and that occurred only in ophthalmic neurones. PMID- 6294235 TI - Modifications of the nuclear envelope of BHK cells after infection with herpes simplex virus type 1. AB - Numerous discrete lesions, which we have termed blebs, appeared in the nucleus of BHK cells 10 to 15 h after infection with herpes simplex virus type 1 (HSV-1). They were formed in the inner portion of the nuclear envelope by the apposition of two thickened lamellae overlying a vacuole. As demonstrated by electron microscopic studies, blebs were regular and associated with the peripheral lamina in the nucleus, averaging 3.5 blebs per micron2. They appeared to be associated with an enrichment of the 155K major capsid protein in the nuclear membrane subfractions as compared with the protein composition of nuclei and plasma membrane fractions. We propose that blebs represent the site of assembly of capsid proteins before DNA insertion and eventual envelopment. PMID- 6294237 TI - Selective blockade of brain alpha 2-autoreceptors by yohimbine: effects on motor activity and on turnover of noradrenaline and dopamine. AB - The motor activity of groups of three mice was increased by yohimbine at doses up to 3 mg/kg intraperitoneally. The turnover of dopamine and noradrenaline in the mouse brain, as assessed by the disappearance of catecholamines following treatment with the tyrosine hydroxylase inhibitor alpha-methyltyrosine, was accelerated by yohimbine with a peak effect after 10 mg/kg intraperitoneally. Prazosin (3 mg/kg i.p.) completely antagonized the stimulatory effect of yohimbine on motor activity and on dopamine turnover but it somewhat potentiated the stimulatory effect on the turnover of noradrenaline. Amphetamine reversed the prazosin-induced hypomotility, indicating that prazosin can selectively block postsynaptic alpha 1-receptors. Yohimbine did not stimulate motor activity following 10 mg/kg and it retarded the turnover of dopamine following 30 mg/kg. These actions might be due to blockade of postsynaptic alpha-receptors by yohimbine. The data indicate that yohimbine at low doses stimulates motor activity and dopamine turnover by selectively blocking alpha 2-autoreceptors leading to increased release of noradrenaline and subsequent activation of post synaptic alpha 1-receptors. PMID- 6294238 TI - A comparative study by serial electron microscopy of neuromuscular junctions in the dimorphic claws of the snapping shrimp, Alpheus heterochelis. AB - We compared the neuromuscular junctions on the main closer muscle in the first pair of chelipeds in the snapping shrimp Alpheus heterochelis by serial section electron microscopy. We sought an ultrastructural basis for the different behavioral and physiological functions of these dimorphic claws and for the role of the nervous system in claw transformation. We were unable to detect any statistically significant morphological differences between the junctions. Further, we found the muscle fiber populations and filament arrangements, as well as the electrical properties of the fibers, to be more homogeneous and similar to each other in A. heterochelis than those reported for another species, A. armillatus. We consider our results in light of recent data on the anatomy and electrical properties of the motor neurons within the CNS and conclude that the neural trigger for claw transformation involves factors not revealed by conventional electron microscopy. PMID- 6294239 TI - Calcium-activated potassium conductance noise in snail neurons. AB - Current fluctuations were measured in small, 3-6 micrometers-diameter patches of soma membrane in bursting neurons of the snail, Helix pomatia. The fluctuations dramatically increased in magnitude with depolarization of the membrane potential under voltage clamp conditions. Two components of conductance noise were identified in the power spectra calculated from the membrane currents. One component had a corner frequency which increased with depolarization. This component was blocked by intracellular injection of TEA and was relatively insensitive to extracellular calcium levels (as long as the total number of effective divalent cations remained constant). It was identified as fluctuations of the voltage-dependent component of delayed outward current. The second component of conductance noise had a corner frequency which decreased with depolarization. It was relatively unaffected by TEA injection and was reversibly blocked by substitution of extracellular calcium with magnesium, cobalt, or nickel. This second component of noise was identified as fluctuations of the calcium-dependent potassium current. The results suggest that the two components of delayed outward current are conducted through physically distinct channels. PMID- 6294240 TI - Swelling of the Muller fibers in the chicken retina. AB - A high potassium concentration (33 meq) in the solution superfusing the isolated chicken retina causes an increase in the tissue transparency. An L-glutamate (1 mM) or L-proline (10 mM) solution has the same effect. Swelling of the Muller fibers, which have a radial position in the retina, could explain the transparency increase. This possibility was investigated in electron micrographs of retinas subjected to these treatments and fixed by freeze-substitution to preserve the water distribution in the tissue. The Muller fibers in the controls had a mean diameter of 0.22 micrometer. The fibers in retinas bathed for 2 min in the high [K+] solution were more than three times as thick (0.74 micrometer); the fibers in glutamate-treated retinas were more than twice as thick (0.49 micrometer). The fibers in the proline-treated retinas had a diameter of 0.39 micrometer. The glutamate- and proline-induced swelling may be due to a K+ release from neuronal elements, acting on the Muller fibers. The fiber swelling was postulated to be the expression of different Donnan equilibriums of fibers bathed in solutions of different K+ concentrations. The observed swelling caused by the high [K+] solution was compared with the theoretical swelling of the fiber as an ideal Donnan system, postulating permeabilities for different ions of the fiber membrane. This suggested that the high [K+] solution causes an increase in Na+ permeability in addition to the permeability of the membrane for K+, Cl-, and HCO3-. Chemical fixation with glutaraldehyde and formaldehyde in an Na-phosphate buffer yielded micrographs in which the Muller fibers of retinas treated with a high [K+] or a glutamate solution had diameters similar to those of the control preparations. PMID- 6294241 TI - Structural and electrotonic connections between developing moth muscle fibers. PMID- 6294242 TI - Formation of hydroxyl radicals from the paraquat radical cation, demonstrated by a highly specific gas chromatographic technique. the role of superoxide radical anion, hydrogen peroxide, and glutathione reductase. AB - Yeast glutathione reductase catalyzes an NADPH-dependent reduction of the herbicide paraquat in vitro. The single-electron reduced paraquat radical reacts with O2 to generate the superoxide radical, O2.-. Hydroxyl radicals (OH.) can also be detected in this assay system by their reaction with phenol to form diphenols, as assayed quantitatively by a highly specific and sensitive method employing gas-liquid chromatography. Formation of hydroxyl radicals can be virtually completely suppressed by catalase and partially suppressed by superoxide dismutase. The role of hydroxyl radicals and superoxide in paraquat toxicity in vivo is discussed. PMID- 6294243 TI - Carbonic anhydrase, 5'-nucleotidase, and 2',3'-cyclic nucleotide-3' phosphodiesterase activities in oligodendrocytes, astrocytes, and neurons isolated from the brains of developing rats. AB - The activities of three myelin-associated enzymes, carbonic anhydrase, 5' nucleotidase, and 2',3'-cyclic nucleotide-3'-phosphodiesterase (CNP), were measured in oligodendrocytes, neurons, and astrocytes isolated from the brain of rats 10, 20, 60, and 120 days old. The carbonic anhydrase specific activity in oligodendrocytes was three- to fivefold higher than that in brain homogenates at each age, and, at all the ages, low activities of this enzyme were measured in neurons and astrocytes. The oligodendrocytes and astrocytes from the brains of rats at all ages had higher activities of the membrane-bound enzyme 5' nucleotidase than was observed in neurons. In oligodendrocytes from 10- and 20 day-old rats, the 5'-nucleotidase activity was two-to threefold the activity in the homogenates (i.e., relative specific activity = 2.0-3.0), and the relative specific activity of this enzyme in the oligodendrocytes declined to less than 1.0 at the later ages, concomitant with the accumulation of 5'-nucleotidase in myelin. The CNP activity was always higher in oligodendrocytes than in neurons, but not appreciably different from that in astrocytes from 20 days of age onward. The relative specific activity of CNP was highest in the oligodendrocytes from 10 day-old rats but was lower, at all ages, than we had observed in bovine oligodendrocytes. These enzyme activities in oligodendroglia are quite different in amount and developmental pattern from those reported previously for myelin. PMID- 6294244 TI - Regulation of glycogen metabolism in primary and transformed astrocytes in vitro. AB - Glycogen metabolism was studied in primary and Herpesvirus-transformed cultures of neonatal rat brain astrocytes. A small fraction of the glucose consumed was conserved in glycogen in both the primary and the transformed astrocytic cell cultures. After addition of culture medium containing 5.5 mM glucose, glycogen increased to maximal levels within 2.5 h, the approximate time at which half of the medium glucose was consumed, and rapidly declined thereafter in both the primary and transformed astrocytic cultures. Maximum levels of glycogen were apparently related to the cell density of the Herpesvirus-transformed cultures, but primary cultures did not show this behavior. At any given cell density, maximal levels of glycogen were dependent on the concentration of extracellular glucose. Administration of glucose caused a transient activation of glycogen synthase alpha and a rapid inactivation of glycogen phosphorylase alpha. PMID- 6294245 TI - Regulation of glycogenolysis in transformed astrocytes in vitro. AB - Cultured astrocytes, transformed by Herpesvirus, were used as a model system to study several aspects of the control of glycogenolysis. Adrenergic agonists such as norepinephrine and isoproterenol caused an immediate and dose-dependent increase in the intracellular levels of cyclic AMP. Concomitant with the initial phase of cyclic AMP increase, conversion of phosphorylase b to a and glycogenolysis were observed. The elevation of cyclic AMP, phosphorylase conversion, and glycogenolysis were simultaneously blocked by beta-adrenergic blockers, but not by alpha-adrenergic blocking agents. Repeated administration of norepinephrine caused an attenuated response in both cyclic AMP accumulation and glycogenolysis. Glycogen degradation is also partially regulated by glucose availability. In the presence of glucose, norepinephrine-induced glycogenolysis is blocked, despite elevations in cyclic AMP. The direct role of glucose is postulated, since glucose analogs mimic the effects of glucose. PMID- 6294246 TI - Changes of beta-endorphin and Met-enkephalin content in the hypothalamus pituitary axis induced by aging. AB - The amounts of beta-endorphin- and Met-enkephalin-immunoreactive material are higher in the pituitary of aged rats. However, the aging process decreases the content of beta-endorphin-, but does not affect that of Met-enkephalin immunoreactive material, in hypothalamus. Thus, it seems that the regulatory mechanisms in the two areas are differentially affected by increasing age. On the other hand, the pituitary increase of these peptides is in line with the assumption that in the elderly the hormonal response to stress is impaired. PMID- 6294247 TI - Characterization of the solubilized GABA and benzodiazepine receptors from various regions of bovine brain. AB - GABA and benzodiazepine receptors were solubilized from bovine cerebral cortex, cerebellum, and hippocampus and then partially purified by gel filtration and characterized. The apparent molecular weights of all these receptors were determined to be 600,000-650,000 by gel filtration, the sedimentation coefficients being 11.0-11.3 S by sucrose density gradient centrifugation. [3H]Muscimol was bound to two classes of sites in fractions from all three regions, and [3H]flunitrazepam bound to one class of sites. A comparison of the ratios of Bmax for flunitrazepam binding to Bmax for muscimol binding revealed that the fractions from the hippocampus exhibited a much higher ratio of benzodiazepine binding sites than were detected in fractions from the cortex and cerebellum. GABA agonist and antagonist inhibited [3H]muscimol binding to the fractions from these regions, at similar concentrations. Benzodiazepine agonists and antagonists also inhibited [3H]flunitrazepam binding in these three fractions, with similar potency. CL 218,872, however, inhibited [3H]flunitrazepam binding in the cerebellar fraction with the lowest IC50 value and that in th hippocampal fraction with the highest IC50 value. Hill coefficients for CL 218,872 inhibition were 0.98, 0.64, and 0.58 for cerebellum, cortex, and hippocampus, respectively. PMID- 6294248 TI - Attenuation of enkephalin activity in neuroblastoma X glioma NG108-15 hybrid cells by phospholipases. AB - The role of membrane phospholipids in enkephalin receptor-mediated inhibition of adenylate cyclase (EC 4.6.1.1) activity in neuroblastoma X glioma NG108-15 hybrids was studied by selective hydrolysis of lipids with phospholipases. When NG108-15 cells were treated with phospholipase C from Clostridium welchii at 37 degrees C, an enzyme concentration--dependent decrease in adenylate cyclase activity was observed. The basal and prostaglandin E1 (PGE1)-stimulated adenylate cyclase activities were more sensitive to phospholipase C (EC 3.1.4.3) treatment than were the NaF-5'-guanylylimidodiphosphate (Gpp(NH)p)-sensitive adenylate cyclase activities. Further, Leu5-enkephalin inhibition of basal or PGE1 stimulated adenylate cyclase activity was attenuated by phospholipase C treatment, characterized by a decrease of enkephalin potency and of maximal inhibitory level. [3H]D-Ala2-Met5-enkephalinamide binding revealed a decrease in receptor affinity with no measurable reduction in number of binding sites after phospholipase C treatment. Although opiate receptor was still under the regulation of guanine nucleotide after phospholipase C treatment, adenylate cyclase activity was more sensitive to the stimulation of Gpp(NH)p. Thus, the reduction of opiate agonist affinity was not due to the uncoupling of opiate receptor from N-component. Further, treatment of NG108-15 hybrid cell membrane with phospholipase C at 24 degrees C produced analogous attenuation of enkephalin potency and efficacy without alteration in receptor binding. The reduction in enkephalin potency could be reversed by treating NG108-15 membrane with phosphatidylcholine, but not with phosphatidylserine, phosphatidylinositol, or cerebroside sulfate. The enkephalin activity in NG108-15 cells was not altered by treating the cells with phospholipase A2 o phospholipase C from Bacillus cereus. Hence, apparently, there was a specific lipid dependency in enkephalin inhibition of adenylate cyclase activity. PMID- 6294249 TI - Enkephalins and opiate antagonists control calmodulin distribution in neuroblastoma-glioma cells. AB - The calcium binding protein calmodulin and the opiate receptor binding sites are unevenly distributed in various subcellular fractions of neuroblastoma-glioma NG108-15 cells. The crude mitochondrial-membrane fraction of these cells contains two membrane fractions that are separable by sucrose gradient centrifugation. These two differ in the content of both calmodulin and opiate receptors. Leucine enkephalin and D-Ala2-methionine enkephalinamide decrease the amount of membrane bound calmodulin in the NC108-15 cells in a time- and dose-dependent manner, whereas the opiate antagonists naloxone and levallorphan have an opposite effect. Naloxone blocks the effect of leucine enkephalin and dextrallorphan has no significant effect. The opiate alkaloids entorphine and phenazocine induce changes similar to that of the enkephalins whereas morphine is inactive even at high concentrations. The alteration in the amount of membrane-bound calmodulin after a short incubation (15 min) with the enkephalins or with naloxone is reflected as an opposite change in the amount of calmodulin in the cell cytosol. Naloxone and levallorphan also increase the number of opiate receptors in NG108 15 cells but dextrallorphan has no such effect. Modulation of the intracellular distribution of calmodulin by opioid peptides and alkaloids may control the activity of various membrane-bound and cytosolic systems that are calmodulin- and/or calcium-dependent. PMID- 6294250 TI - The effects of the pyrethroids deltamethrin and cismethrin on nerve excitability in rats. AB - Deltamethrin produced a prolonged period of increased excitability following the passage of a nerve impulse, which was dose-related and lasted up to 400 ms. Excitability changes were detected without neurological signs following a single IV injection of 0.5 mg/kg and feeding 200,100 and 50 ppm in the diet for up to 8 weeks. No changes were detected following 0.3 mg/kg IV or 25 ppm in the diet. No cumulative effects were detected during chronic feeding. Cismethrin produced increased nerve excitability only between 2 and 4 ms after a nerve impulse. Excitability changes after cismethrin were biphasic during the first 20 ms. PMID- 6294251 TI - Adult ceroid-lipofuscinosis: diagnostic value of biopsies and of neurophysiological investigations. AB - In a sibship of ten, three brothers presented with an adult form of ceroid lipofuscinosis. The diagnosis was confirmed by necropsy of the first patient and was made by electron microscopy of eccrine sweat glands and of skeletal muscles in the two others. Somatosensory evoked potentials were characterised by biphasic, nearly monophasic, very high voltage complexes totally unlike those found in normal controls. Similar sensory evoked potentials were however recorded in other types of ceroid-lipofuscinosis. While electron microscopy of easily available tissues gives fairly specific results, sensory evoked potentials can bring supportive diagnostic evidence in adult ceroid-lipofuscinosis. PMID- 6294252 TI - Association of Klippel-Trenaunay-Weber syndrome with myotonic dystrophy. PMID- 6294253 TI - Summation of rapid tactile stimuli in parietal lobe disease. AB - The perception threshold for trains of rapid tactile pulses, applied to the index finger, has been measured in patients with parietal lobe lesions and in patients with median nerve lesions. The former patients had increased perception thresholds for single tactile pulses on the abnormal side. With successively prolonged pulse trains, the threshold decreased exponentially to reach a stable level after 150-400 ms. In contrast, the median nerve patients had increased perception thresholds for tactile pulses irrespective of pulse train duration. PMID- 6294254 TI - Familial granulovacuolar lobular myopathy with electrical myotonia. AB - We report 3 patients with a myopathy characterized by profound selective muscle wasting and weakness, electrical myotonia without clinical myotonia and an unusual muscle biopsy. Cryostat sections showed muscle fibers with vacuoles containing hematoxylinophilic granules, and 30% of type I fibers showed demarcation of their sarcoplasm into "lobules" due apparently to reorganization of myofibrillar elements. The electrical myotonia suggests an underlying muscle membrane defect. Two of the patients are siblings suggesting that the disorder may be inherited. PMID- 6294255 TI - A longitudinal study of changes in motor units in motor neuron disease. AB - In a longitudinal investigation of 14 patients with motor neuron disease motor units were studied in the biceps and first dorsal interosseous muscles in both arms, using single fibre EMG. The fibre density usually increased initially and this was occasionally accompanied by temporary improvement in strength. Later, as the fibre density decreased, increasing weakness and atrophy developed. The fibre density was rarely as high at any stage of the disease as in other chronic neurogenic disorders. Marked asymmetry was observed in individual patients at all stages of the disease. PMID- 6294256 TI - Binding of herpes simplex virus to regional areas of the rodent brain. AB - The binding of 3H-thymidine labeled herpes simplex virus to homogenates of rodent brain regions and liver was examined. The results indicated that binding sites in the brain are distributed in a non-uniform manner, the specific pattern observed depending upon the age of the animal. In addition, binding was found to be reduced by pretreatment of the labeled virus with specific antiserum or trypsin. These results suggest that the distribution of herpes simplex virus receptors is one of the factors determining neurotropism and the localization of infection within the brain. PMID- 6294257 TI - Neurophysiological studies in adrenomyeloneuropathy. A report on five cases. AB - Neurophysiological studies were performed in 5 patients in two families suffering from adrenomyeloneuropathy (AMN). The diagnosis was supported by electron microscopy of nerve twigs in all cases and by the demonstration in 2 cases of increased levels of saturated very long chain fatty acids in cultured fibroblasts (Moser, personal communication). Measurements of the sensory-motor conduction velocities demonstrated the variability of the peripheral nerve damage in AMN, further confirmed by quantitative studies of sensory nerve biopsies. Somatosensory evoked potentials (SEP) were abnormally delayed and their configuration was abnormal, mainly following stimuli applied to the lower limbs. Our data suggest a more severe involvement of the fasciculus gracilis and an extension of the lesions to the medial lemnisci in agreement with the few postmortem reports showing multifocal demyelination. Brain stem auditory evoked potentials (BAEP) were delayed, pointing towards lesions between cochlear nerve and superior olivary nucleus and also at lateral lemniscal level. Morphological confirmation is lacking but the close topographical relationship between the secondary auditory pathways and the medial lemnisci indicates that even small lesions could damage simultaneously both pathways. Neurophysiological studies contribute to the diagnosis of AMN, they confirm the inter- and intrafamilial variability of the clinical features and help to explain the signs and symptoms of this condition. PMID- 6294258 TI - Normal platelet aggregation in myotonic dystrophy. AB - In 18 patients with myotonic dystrophy, spontaneous aggregation and platelet aggregation induced by thrombin, adenosine diphosphate and epinephrine were compared with normal aggregation patterns. In 17 of the 18 patients the results were not significantly different from normal. In 1 patient spontaneous aggregation and hypersensitive platelets were found. These results are in disagreement with earlier reports on a specific hypersensitivity to epinephrine in myotonic dystrophy. Neither the clinical data (myotonia, paresis) nor the laboratory data (creatine kinase, myoglobin, immunoglobulin G) were correlated with the platelet aggregations. PMID- 6294259 TI - Monosynaptic projections of individual spindle group II afferents to type identified medial gastrocnemius motoneurons in the cat. PMID- 6294260 TI - Computerized tomographic and pathologic studies of the untreated, quiescent, and recurrent glioblastoma multiforme. AB - Pathological findings in 20 cases of glioblastoma multiforme were correlated with clinical histories and computerized tomographic (CT) scans. This was done to define the neoplasm in three stages: before treatment, during remission, and during recurrence. The untreated lesions were markedly cellular neoplasms composed predominantly of small anaplastic cells. The radiographic central region of low density was necrosis, the enhancing rim was a cellular zone of viable neoplasm, and the perilesional low-density area was edema with infiltrating tumor. In these 20 cases, all of the identifiable neoplasms lay within the zone of peritumoral edema or contrast enhancement, although small anaplastic cells may have been present in more distant regions. The lesions in remission were remarkable for their minimal mass effect, discrete nature, extensive necrosis, and content of large bizarre glia. The large cells were confined to the original tumor bed and were consistent with neoplastic cells inactivated and immobilized by radio- and chemotherapy. These lesions were accurately localized by CT scanning. The recurrent lesions were heterogeneous, but most were formed of widely disseminated small anaplastic cells. The highly cellular regions of such lesions could be localized by CT scanning, but CT could not detect less cellular foci in the cerebrum, cerebellum, or brain stem. In one patient, the contrast enhancing lesions of "recurrence," were foci of radionecrosis, underscoring the difficulty in distinguishing this entity from recurrent neoplasm. PMID- 6294261 TI - Comparison of suture methods and materials in experimental inferior alveolar nerve grafting. AB - Currently, peripheral nerve defects are being treated successfully by microsurgical techniques using autogenous nerve transplantations and synthetic suture materials. This paper describes the influence of various suture methods and materials on nerve repair as experimentally evaluated in rabbits. The results show that the action potentials of the nerves anastomosed with polyglycolic acid sutures recovered faster than those anastomosed with nylon. PMID- 6294262 TI - An ultrastructural study of the origin and function of basophilic degeneration in human cardiac muscle--cardiac colloid. Type 1. AB - The myofibres in biopsies obtained from the left ventricle during cardiopulmonary bypass, in patients undergoing surgery for mitral and aortic valve replacement and the correction of atrial septal defect, contained inclusions thought to be type 1 cardiac colloid. Electron microscopy showed this material to be synthesised by and contained within a grossly dilated rough endoplasmic reticulum. The presence of membrane-bound aggregates of non-fibrillar sarcoplasmic organelles within colloid deposits, together with osmiophilic inclusions thought to be related to lipofuscin, both within and moving from this structure, suggest that type 1 cardiac colloid is an autophagic vacuole designed to degrade ageing or redundant sarcoplasmic organelles. PMID- 6294263 TI - Single-dose ceftriaxone pharmacokinetics in pediatric patients with central nervous system infections. AB - Ceftriaxone has greater in vitro and in vivo efficacy against many common bacteria than other third-generation cephalosporins. Single-dose ceftriaxone pharmacokinetics were studied in 17 patients, aged 0.6 to 52 months, with infections of the central nervous system. Patients received a randomized dose of 50 or 75 mg/kg ceftriaxone intravenously over 5 minutes on the second to fifth day of illness. Serial blood samples were collected over 24 hours in all patients, and cerebrospinal fluid (CSF) was obtained 1 to 4.5 hours after injection. Ceftriaxone mean peak plasma concentrations, determined by high-power liquid chromatography, were 267 and 184 microgram/ml for the 75 and 50 mg/kg dosage groups, respectively. The harmonic mean elimination half-life was 4.2 hours, and the mean percent drug penetrance into CSF was 4.8 +/- 3.5%. Of CSF studies evaluated, the glucose concentration was correlated most closely (inversely) with CSF penetration of ceftriaxone. Individual CSF concentrations of ceftriaxone exceeded the minimal inhibitory concentrations of the respective bacteria causing infection by 480 to 5,600 times. Ceftriaxone may be useful in the treatment of serious pediatric infections, including meningitis. PMID- 6294264 TI - Sodium valproate and corticotropin suppression in the child treated for seizures. PMID- 6294265 TI - Epstein-Barr virus and infantile papular acrodermatitis. PMID- 6294266 TI - Renin substrate depletion in salt-losing congenital virilizing adrenal hyperplasia: low plasma renin activity despite increased renin concentration. PMID- 6294267 TI - Intracardiac Wilms' tumor: diagnosis and management. AB - Complete excision of Wilms' tumor may require resection of adjacent organs and removal of intracaval tumor propagation. Extension of tumor to the right atrium can be determined preoperatively guiding a direct and safe approach to intracardiac tumor at the time of nephrectomy. Preoperative ultrasonography of two children with Wilms' tumor demonstrated caval and right atrial tumor. Laparotomy for nephrectomy and abdominal caval exposure was combined with cardiopulmonary bypass and atriotomy. In both patients, tumor contiguous with the renal pelvis extended from the iliac bifurcation into the right atrium with a large atrial mass. In one patient nephrectomy was performed first, and she was then placed on cardiopulmonary bypass. Caval tumor was easily removed through the atriotomy and open renal vein. In the second patient, bypass was instituted first because of cardiac instability. The large right atrial mass extended through an atrial septal defect into the left atrium. The cardiac tumor and a large amount of caval tumor were removed. Bypass was discontinued after repair of the ASD. Tumor remained in the IVC below the renal veins necessitating a separate venotomy. Combined abdominal and cardiac exploration allows safe and complete excision of all gross tumor. Ultrasonography is a sensitive and noninvasive method of diagnosing retrohepatic and atrial tumor extension and can be obtained easily even on very sick patients. PMID- 6294268 TI - The abdominal technetium scan (a decade of experience). AB - Out of 270 children with gastrointestinal symptoms, the indications for technitium scanning were: gastrointestinal tract bleeding (165 patients), abdominal pain (99 patients) and a history of intussusception (6 patients). Thirty children had abnormal findings, while the remaining 240 patients had "normal" scans. Four of the 30 children with positive scans were not explored, while the others underwent laparotomy. Of the 26 operated patients, 12 (46%) had a Meckel's diverticulum. Nine patients (34%) had other pathologic lesions that were detected by the scan. Five had true "false positives" as no pathologic lesions were found. Of the 240 children with negative scans, 19 were eventually explored because of persistent symptoms or clinical findings. Two of these had a Meckel's diverticulum. Eleven had a negative exploration while six had other surgical lesions. Technitium scan should reliably detect around 80%-90% of Meckel's diverticula. It will also accurately exclude the diagnosis of Meckel's diverticulum in over 90% of patients. PMID- 6294269 TI - [Quantum chemical studies on enzymatic reaction mechanisms]. PMID- 6294270 TI - [Effects of total saponins extracted from several crude drugs on rat adrenocortical hormone secretion]. PMID- 6294271 TI - [Chemistry for the basis of biological activities of macrocyclic polyamines]. PMID- 6294272 TI - Effects of nitroprusside and 8-bromo-cyclic GMP on the contractile activity of the rat aorta. PMID- 6294274 TI - N-Ethylmaleimide-induced alteration in the interaction of agonists with muscarinic cholinergic receptors of rat brain. PMID- 6294273 TI - 45Ca fluxes in isolated toad bladder epithelial cells: effects of agents which alter water or sodium transport. AB - Vasopressin enhances osmotic water flow and sodium transport across the toad urinary bladder by mechanisms involving cyclic AMP and calcium. It is believed that changes in intracellular calcium concentration or in its binding to membranes may in part mediate the effects of vasopressin. In addition, several agents which alter the response of the toad bladder to vasopressin may also act by altering cellular calcium metabolism. The effects of vasopressin and several agents which modify its effects in the toad bladder were studied on 45Ca fluxes in isolated epithelial cells from the toad bladder. Compartmental analysis of 45Ca exchange revealed three components. Vasopressin reduced the amount of calcium in the most rapidly exchanging pool from 1.67 +/- 0.20 to 0.86 +/- 0.12 nmol/mg of protein (P less than .025) and the most slowly exchanging pool from 2.72 +/- 0.26 to 1.90 +/- 0.34 nmol/mg of protein (P less than .001), while not affecting the intermediate pool. Theophylline, which mimics the natriferic and hydroosmotic effects of vasopressin, also mimicked the effects on 45Ca exchange by vasopressin. Exogenous cyclic AMP and the prostaglandin endoperoxide analog U46619, which mimic the hydroosmotic effect of vasopressin, also reduced the amount of calcium in the most slowly exchanging pool. Prostaglandin E1, which inhibits the hydroosmotic effect of vasopressin at the concentrations used increased the size of the most slowly exchanging pool. These studies suggest that prostaglandins and other agents which alter the effect of vasopressin in the isolated toad bladder may elicit their effects in part by influencing the calcium concentration at some critical site. PMID- 6294275 TI - Botulinum neurotoxin type E: studies on mechanism of action and on structure activity relationships. AB - Single chain type E botulinum neurotoxin was isolated from culture fluids of Clostridium botulinum (strain Alaska E-43). The neurotoxin, which migrated as a single band in polyacrylamide gel electrophoresis with sodium dodecylsulfate, had a molecular weight of approximately 147,000. Single chain type E neurotoxin that was exposed to trypsin was converted to a dichain molecule. Pretreatment of the single chain molecule with 1,2-cyclohexanedione, a reagent that selectively modifies arginine residues, inhibited trypsin-induced generation of the dichain molecule. In dose-response experiments (10(-13) to 10(-9) M) on the isolated neuromuscular junction (phrenic nerve-hemidiaphragm preparation), the dichain neurotoxin was approximately two orders of magnitude more potent than the single chain neurotoxin. Neither specie of neurotoxin (1 pmol/mouse, in vivo; 1 X 10( 11) M, in vitro) was very effective in blocking autonomic transmission (vagus nerve-atrium preparation). The neuromuscular blocking action of the dichain molecule was divided into a sequence of three steps. There was an initial binding step that was relatively rapid, little influenced by temperature and which left the neurotoxin partially accessible to the neutralizing effects of antitoxin. There was a translocation step that was temperature dependent, antagonized by ammonium chloride and methylamine hydrochloride and which caused the neurotoxin to become inaccessible to the neutralizing effects of antitoxin. Finally, there was an intracellular lytic step, during which the toxin blocked excitation secretion coupling. PMID- 6294276 TI - Characterization of adrenergic receptor binding in rat lung: physiological regulation. AB - The ontogeny, circadian rhythm and sex differences of adrenergic receptors were studied in a crude particulate fraction prepared from rat lung. The density of alpha-2 adrenergic receptors ([3H]yohimbine binding sites) decreased rapidly with age from 304 fmol/mg of protein on the 1st day to 123 fmol/mg after the 1st week and then to an undetectable level at 5 weeks of age. Alpha-1 adrenergic receptors ([3H]prazosin binding) showed a moderate variation in density, with the greatest density at the 3rd week of age (311 fmol/mg). The density at birth (194 fmol/mg) was higher than the adult (126 fmol/mg). Beta adrenergic receptors ([3H]dihydroalprenolol binding) demonstrated changes in density with a sharp rise in the 3rd week (to 813 fmol/mg from 315 fmol/mg at birth), which was maintained through the 10th week. Unlike receptor densities, the affinity constants were not significantly altered during postnatal development. Alpha-1 and beta receptors in the adult rat lung did not show any significant changes in their densities or their affinities in the two other physiological variables studied (circadian rhythm and sex). During postnatal development, the highest concentration of norepinephrine in the rat lung is reported to be at about 3 weeks of age, which correlates with the changes in both alpha-1 and beta adrenergic receptors, but not alpha-2 receptors. The presence of alpha-2 receptors in the lungs of young, but not mature animals, may suggest an important developmental role, which is not yet understood. PMID- 6294277 TI - Interactions of salsolinol and its mono-O-methylated analogs with adrenergic and dopaminergic receptors in the rabbit ear artery. AB - We have examined the action of the tetrahydroisoquinoline derivatives salsolinol, 6-O-methyl salsolinol (6-O-Me-Sal) and 7-O-methyl salsolinol (7-O-Me-Sal) on adrenergic and dopaminergic receptors in the isolated and perfused rabbit ear artery. The racemic form of the three compounds and the S-(--)-isomer of 6-O-Me Sal were used. Salsolinol (0.3-10 micrometers) produced concentration-dependent inhibition of the vasoconstrictor response to electrical stimulation of the periarterial sympathetic nerves but did not inhibit the vasoconstrictor response to exogenous norepinephrine. The inhibitory effect of salsolinol on neurotransmission was antagonized by yohimbine, but not by sulpiride or propranolol. The dissociation constant (KB) for yohimbine acting as an antagonist of salsolinol was 98 +/- 8 nM. 6-O-Me-Sal and 7-O-Me-Sal did not affect the response to nerve stimulation or norepinephrine administration; however, these mono-O-methylated analogs of salsolinol antagonized the inhibition of neurotransmission produced by dopamine. The KB values for 6-O-Me-Sal and 7-O-Me Sal acting as antagonists of dopamine were 1.3 +/- 0.2 and 6.4 +/- 0.31 microM, respectively. S-(--)-6-O-Me-Sal, with a KB value of 0.64 +/- 0.06 microM, was about twice as potent as racemic 6-O-Me-Sal. We conclude that salsolinol acts as an agonist on prejunctional alpha adrenergic receptors and that 6-O-Me-Sal and 7 O-Me-Sal act as antagonists on dopaminergic receptors. PMID- 6294278 TI - Effect of yohimbine on rat prolactin secretion. AB - It was recently proposed that yohimbine (YOH), an indole alkaloid with multiple pharmacological effects, is an antagonist of the D2 dopamine (DA) receptor. Because the pituitary DA receptor involved in the inhibition of prolactin (PRL) secretion is the prototypic D2 receptor, we examined the effect of YOH on PRL secretion in male rats. YOH produced marked, dose-dependent and sustained increases in plasma PRL levels. However, YOH did not block the inhibitory effect of DA on PRL release from rat pituitary glands in vitro, did not displace [3H]spiperone from bovine pituitary membranes and had no effect on the concentration of DA in pituitary stalk plasma of anesthetized rats, suggesting that the stimulation of PRL release by YOH is not due to its antidopaminergic effects. Clonidine, an alpha-2 adrenergic agonist, produced a partial, non-dose dependent inhibition of the YOH-induced rise in serum PRL levels. Two antagonists of the H1 histamine receptor, diphenhydramine and promethazine, markedly antagonized the PRL-releasing effect of YOH, but another H1 blocker, chlorpheniramine, and an H2 antagonist, metiamide, had no effect. Serotonin receptor blockers, cyproheptadine, mianserin and pizotifen, and the opiate antagonist, naloxone, also had no effect on the PRL response to YOH. Nevertheless, the PRL-releasing effect of YOH was potentiated 24 hr after the administration of reserpine or para-chlorophenylalanine, an inhibitor of serotonin synthesis. Thus, the mechanisms by which YOH stimulates rat PRL secretion has has not been fully elucidated. It is possible that YOH may stimulate PRL secretion by a novel mechanism, possibly through the intervention of a PRL-releasing factor. PMID- 6294279 TI - Postjunctional supersensitivity of the smooth muscle of the rat vas deferens induced by calmodulin-antagonizing drugs applied locally to the hypogastric plexus. AB - Local application of 100 micrograms of chlorpromazine hydrochloride (CPZ) to the hypogastric plexus of the rat produced supersensitivity of the smooth muscle of the vas deferens. In comparison with surgical denervation, CPZ produced a similar increase in the maximum response but less leftward shift of the dose-response curves to norepinephrine. The changes in dose response curves to methacholine and potassium induced by CPZ were substantially similar to those induced by denervation. Virtually identical supersensitivity was brought about in the rat vas deferens by the same treatment with other drugs which possess calmodulin antagonizing actions. CPZ produced no change in norepinephrine content of the tissue. Contractile responses of the tissue to tyramine were markedly potentiated by CPZ, but the responses to nerve stimulation were not significantly altered except for that at the lowest frequency. In electrophysiologic studies, while spontaneous junction potentials could be detected in all of the impaled cells, they occurred with lesser frequency and smaller amplitude in all of the impaled cells in the CPZ-treated rat vas deferens than in the control tissue. Nevertheless, the junction and action potentials could be induced by nerve stimulation in all smooth muscle cells. The results indicate that CPZ and also other drugs used in the present study produced postjunctional supersensitivity accompanied by a large increase in the maximum response, but almost no prejunctional supersensitivity in the rat vas deferens. PMID- 6294280 TI - Inhibition of ouabain-induced increase in Na content of cultured myocardial cells by quinidine and procainamide. AB - Addition of ouabain caused gradual increases of both the Na content of cultured myocardial cells and the rate of Ca++ uptake by the cells. Ouabain-induced irregular beating of the cells (ouabain toxicity) appeared to develop when the Na content and the rate of Ca++ uptake exceeded about 1.5 and 2.0 times, respectively, the normal levels. Quinidine and procainamide prevented ouabain induced increases of the Na content and the rate of Ca++ uptake as well as ouabain-induced toxicity. The problem of how quinidine and procainamide counteract the effects of ouabain was then studied. Quinidine and procainamide did not affect the Na+-Ca++ exchange activity. Na+,K+-adenosine triphosphatase activity, Na+-pumping out activity or ouabain-binding activity of myocardial cells, but inhibited passive Na+ influx, which is achieved by a simple diffusion system. From these observations, it is suggested that inhibition by quinidine or procainamide of passive Na+ influx indirectly prevents ouabain-induced increase in the intracellular Na content of myocardial cells and that this presumably explains at least in part the inhibitory effect of quinidine and procainamide on ouabain-induced irregular beating. PMID- 6294281 TI - Pharmacological characterization of purinergic receptors in the rat vas deferens. AB - Using the isolated rat vas deferens, we have confirmed the existence of P1 purinergic receptors whose activation results in an inhibition of the neurogenic twitch of the vas deferens. The observed order of potency for agonists (adenosine ethyl carboxamide greater than 2-chloroadenosine greater than adenosine greater than 5'-AMP greater than 5'-ADP greater than ATP) and antagonism of these effects by theophylline supports a P1-mediated response. Metabolically stable analogs of ATP elicited dose-dependent contractile responses which were quantitatively greater than, but qualitatively comparable to, ATP-induced responses. The order of potency for the eliciting contraction was the following: adenylyl-5 imidodiphosphate = beta-gamma-methylene ATP greater than adenosine tetraphosphate much greater than ATP greater than ADP. Interestingly, these compounds also produced an inhibition of the neurogenic twitch with a similar rank order of potency. This response was not due to the activation of P1 receptors insofar as high concentrations of theophylline failed to attenuate either the inhibition of the neurogenic twitch or the contractile response induced by these agonists. Thus, these data demonstrate the presence of both P1 and P2 purinergic receptors in the rat vas deferens. In addition, the data are consistent with the idea that two distinct classes of P2 receptors exist in this tissue. Furthermore, these data suggest that the rat vas deferens provides a useful tissue for studying compounds which interact with both major subtypes of purinergic receptors. PMID- 6294282 TI - Vascular effects of the stereoisomers of dobutamine. PMID- 6294283 TI - Inhibition of tail-flick and shaking responses by intrathecal and intraventricular D-Ala2-D-Leu5-enkephalin and beta-endorphin in anesthetized rats. AB - The present study compares the potencies of D-Ala2-D-Leu5-enkephalin (DADL) and beta-endorphin (beta-EP) injection intrathecally and i.c.v. on the inhibition of tail-flick and ice water-induced shaking responses in pentobarbital-anesthetized rats. Peptides were injected stereotaxically into the third ventricle or into the spinal subarachnoid space. The tail-flick response was measured 10 or 20 min after the injection of DADL or beta-EP, respectively, after which the rats were immersed for 5 min in ice water and the number of shakes was counted. Intrathecal DADL and beta-EP were equipotent in inhibiting both the tail-flick and shaking responses. However, treatment with naloxone (2 mg/kg i.p.) shifted the dose response curve for both responses to the right more for the beta-EP than for DADL. Unlike the equipotency intrathecally, DADL by i.c.v. injection was less potent than beta-EP in inhibiting both the tail-flick and shaking responses. Previous results in the unanesthetized rat had also shown that i.c.v. DADL was less potent than beta-EP in inhibiting the tail-flick response. Thus, the spinal cord is sensitive to both DADL and beta-EP, whereas the supraspinal area is more sensitive to beta-EP. Also, because the potency of i.c.v. beta-EP for inhibiting tail-flick but not shaking was substantially reduced in anesthetized compared to unanesthetized rats, the neuronal substrates involved in inhibition of shaking are different from those of inhibition of the tail-flick. The differential sensitivities to i.c.v. vs. intrathecal administration and the unequal responses to the antagonistic action of naloxone indicate that DADL and beta-EP produce their action through different opioid receptors. PMID- 6294284 TI - A kappa opioid effect: increased urination in the rat. AB - The effects of various opioids (mu agonists, kappa agonists and mixed agonists/antagonists) were determined on urination in the normally dehydrated rat. Opioids considered as kappa agonists (bremazocine, ethylketazocine and ketazocine) produced a marked dose-related increase in urination. The mixed agonists/antagonists (cyclazocine, butorphanol and nalorphine) produced less urination than the kappa agonists, but more than the mu agonists (morphine and l methadone). The mu agonists did not increase urine output compared with controls. The increased urination effect was blocked by opioid antagonists in a potency order which indicated that the effect was due to an action at a kappa opioid receptor. The data suggest the hypothesis that dynorphin, a kappa agonist, acts as an endogenous ligand for an autoreceptor which inhibits the corelease of dynorphin and antidiuretic hormone from the neurohypophysis. This decrease in antidiuretic hormone levels produces the increased urination. Increased urination is a simple in vivo test for studying the actions of compounds at kappa opioid receptors. PMID- 6294285 TI - Inotropic and electrophysiological effects of histamine on human ventricular heart muscle. AB - 1. The effects of histamine were investigated on mechanical and electrophysiological parameters in isolated electrically driven human ventricular papillary muscles. The effects of cimetidine and propranolol on histamine responses were also investigated. 2. The effects of histamine were compared with those of noradrenaline, isoprenaline, dimaprit, a selective H2-receptor agonist, and a cyclic AMP derivative, 8-(4-chlorphenylthio) cyclic AMP. 3. The effects of histamine and dimaprit and the effects of cimetidine on histamine responses were also investigated in guinea-pig right ventricular papillary muscles in order to allow a comparison with human papillary muscles. 4. In human papillary muscles, histamine caused concentration-dependent increases in the force of contraction and reductions in both time-to-peak tension and time-to-half-maximal relaxation. Histamine simultaneously caused distinct changes in the action potential configuration with increases in the height and duration of the plateau phase and an increase in the over-all action potential duration. 5. Noradrenaline and isoprenaline produced similar responses to histamine, as did 8-(4 chlorphenylthio) cyclic AMP, consistent with the view that the effects of histamine as well as the beta-adrenoceptor agonists on human ventricle, were associated with cyclic AMP mediated increases in calcium-dependent slow inward current. PMID- 6294286 TI - Occlusion of rubidium ions by the sodium-potassium pump: its implications for the mechanism of potassium transport. AB - 1. The occlusion of rubidium ions by Na, K-ATPase has been investigated by suspending enzyme prepared from pig kidney outer medulla in media containing low concentrations of (86)Rb, forcing the suspensions rapidly through small columns of cation-exchange resin, and measuring the amounts of radioactivity emerging from the columns.2. When the suspension media contained 2 mM-ATP or ADP, or 15 mM NaCl, the amounts of radioactivity emerging from the columns were greatly (and similarly) reduced, presumably because both nucleotides and sodium ions stabilized the enzyme in the E(1) form. (See p. 19 for definition of E(1) and E(2)). The extra radioactivity carried through the columns when nucleotides and sodium were absent was taken as a measure of the amount of rubidium occluded within the enzyme (in the E(2) form) when it emerged from the resin.3. By varying the flow rate, and therefore the time spent by the enzyme on the resin, and relating this to the amount of radioactivity emerging from the columns, we have been able to estimate the rate constant for the conformational change (E(2) --> E(1)) that allows the occluded rubidium ions to escape. At 20 degrees C, and in the absence of nucleotides, it is about 0.1 S(-1).4. The rate constant for rubidium release was the same in a sodium-containing as in a potassium-containing medium. The opposite effects of sodium and potassium ions on the poise of the equilibrium between the E(1) and the E(2) forms of the enzyme must, therefore, be due solely to opposite effects of these ions on the rate of conversion of E(1) to E(2).5. The rate constant for rubidium release was greatly increased by ATP and by ADP. Both nucleotides appeared to act at low-affinity sites and without phosphorylating the enzyme.6. Orthovanadate, in the presence of magnesium ions, stabilized the enzyme in the occluded-rubidium (E(2)Rb) form.7. Ouabain, in the presence of magnesium ions, prevented the occlusion of rubidium ions.8. We have measured the amount of rubidium occluded by the enzyme as a function of rubidium concentration, and estimate that at saturating rubidium concentrations about three rubidium ions can be occluded per phosphorylation site (or per ouabain binding site).9. We have found that the occluded-rubidium form of the enzyme can also be formed by allowing rubidium ions to catalyse the hydrolysis of phosphoenzyme generated by the addition of ATP to enzyme suspended in a high sodium medium.10. The properties of the occluded-rubidium form of the enzyme, and of the two routes that can lead to its formation, suggest that an analagous occluded-potassium form plays a central role in the transport of potassium ions through the sodium-potassium pump. This hypothesis is supported by a detailed consideration of the probable magnitudes of the rate constants of the individual reactions making up the two routes. PMID- 6294287 TI - The electrogenic sodium pump in guinea-pig ventricular muscle: inhibition of pump current by cardiac glycosides. AB - 1. The inhibition of the electrogenic sodium pump in guinea-pig ventricular muscle by cardiac glycosides was studied with a voltage-clamp technique.2. Superfusion of the preparation with dihydro-ouabain (DHO) produced a reversible depolarization of up to 7 mV. When the membrane potential was clamped to a constant value near the resting potential application of DHO produced a corresponding current change in the inward direction which reached a steady state in less than 1 min.3. The drug-induced current change (I(D)) was found to be the result of a parallel shift of the current-voltage relation. The contributions of a change in extracellular K or intracellular Na to the measured I(D) were shown to be very small. From these findings and the results summarized below it was concluded that I(D) represents the blockage of the electrogenic pump current by DHO and that it is proportional to the number of drug molecules bound to the Na-K ATPase in the intact cell.4. The dependence of I(D) on the concentration of DHO applied (5 x 10(-6)-8 x 10(-4) M) was found to be consistent with the predictions of the law of mass action for reversible one-to-one binding of the drug to the Na K pump under equilibrium conditions. From a Scatchard-type plot the equilibrium dissociation constant (K(D)) of DHO was determined to be 4.6 (+/-2.3) x 10(-5) M.5. The steady-state pump current in the resting preparation was calculated to be 0.81+/-0.26 muA/cm(2). It contributed 6.4+/-0.9 mV to the resting potential in Tyrode solution containing 3 mM-K.6. In the smallest preparations used the measured time course of the onset and decay of I(D) agreed with the chemical kinetics of binding and unbinding calculated for various DHO concentrations. The rate constant of unbinding (k(2)) was found to be 3.4 (+/-0.7) x 10(-2) S(-1) and the average rate constant of binding (k(1)) was 7.4 x 10(2) M(-1) S(-1).7. By comparing the effects of ouabain and DHO in the same preparation the following estimates of the chemical constants of ouabain binding to the Na-K pump were obtained: K(D) approximately 1.5 x 10(-6) M; k(1) approximately 4 x 10(3) M(-1) S(-1); k(2) approximately 6 x 10(-3) S(-1).8. An analysis of the transmembrane movements of Na and K in the steady state showed that the measured pump current density is consistent with a counter-transport of 3 Na and 2 K ions. PMID- 6294288 TI - The absence of neuromuscular transmission failure in sustained maximal voluntary contractions. AB - 1. Muscle mass action potentials (M waves) were evoked by supramaximal single shocks to the ulnar nerve given at 5-10 s intervals throughout sustained isometric maximal voluntary contractions (m.v.c.) of the adductor pollicis and first dorsal interosseous muscles. Both muscles were fatigued simultaneously. Recordings were made from the muscle surface and also intramuscularly. 2. During a maximal contraction lasting for 60 s there was 30-50% loss of force. No decline was observed in intramuscularly recorded M wave amplitude, while the areas of the total and half M wave forms increased due to a slowing in conduction velocity. The area measured over a fixed time period declined. No evidence was obtained that these M wave potentials were contaminated by electrical activity arising in adjacent muscles. The size of the single unit potentials appeared to remain unaltered during maximal voluntary activity. 3. We conclude that neuromuscular block is not a cause of force loss during this type of fatiguing voluntary contraction. PMID- 6294289 TI - The appearance and development of chemosensitivity in Rohon-Beard neurones of the Xenopus spinal cord. AB - 1. We have examined the onset and subsequent development of chemosensitivity in Rohon-Beard neurones from the Xenopus spinal cord. These cells become sensitive to bath-applied gamma-aminobutyric acid (GABA) around stage 25 (early tailbud, about 1 d old), and remain so at least until stage 49 (9 d old). In contrast, a number of other neurotransmitter candidates tested caused no potential or conductance change during the same period.2. We examined ionophoretic dose response relations of the cells at stage 26, a couple of hours after the first acquisition of GABA sensitivity. Sensitivities as high as 450 mV/nC were recorded. Comparable sensitivities were recorded between stages 46-49 (5-9 d old).3. Measurements of ionophoretic sensitivities and input resistances during several periods from stage 26 to maturity show that the underlying conductance change for a given GABA dose is likely to increase steadily during this time. A ;sensitivity index' (ionophoretic sensitivity/input resistance) was calculated, which is low at stage 26, higher at intermediate stages (stages 31-42), and highest for mature cells (stages 46-49; 5-9 d of development).4. The reversal potential of the ionophoretic GABA response is the same at stage 26 (-30 mV) as it is in mature cells. Ion substitution experiments show that Na(+) and K(+), but not Cl(-) or Ca(2+), are involved in the response.5. GABA responses at stage 26 are pharmacologically similar to those of mature cells. The responses are blocked by 10 muM-picrotoxin or curare, and muscimol is an agonist in concentrations as low as 1 muM.6. GABA responses at stage 26 desensitize in a manner similar to that seen for mature cells, either with prolonged bath application of GABA or with repetitive ionophoretic application.7. Nearly half of the cells tested at stage 26 respond to glycine, in concentrations as low as 5 muM. This sensitivity is absent by 3(1/2) d of development.8. The responses of Rohon-Beard neurones to GABA are similar to those of other cells in that they involve a conductance increase, are mimicked by muscimol, and are blocked by picrotoxin. These responses are different in that they do not involve Cl(-) and are blocked by low concentrations of curare.9. Many of the characteristics of GABA receptors, i.e. the reversal potential, desensitization, and pharmacology, are constant during development. However, the sensitivity of the cells to GABA and the spectrum of transmitters to which they are sensitive appear to change. PMID- 6294292 TI - Haemoperitoneum due to traumatic rupture of a hepatoma in a patient with haemochromatosis. PMID- 6294290 TI - M-currents and other potassium currents in bullfrog sympathetic neurones. AB - 1. Bullfrog lumbar sympathetic neurones were voltage-clamped in vitro through twin micro-electrodes. Four different outward (K(+)) currents could be identified: (i) a large sustained voltage-sensitive delayed rectifier current (I(K)) activated at membrane potentials more positive than -25 mV; (ii) a calcium dependent sustained outward current (I(C)) activated at similar positive potentials and peaking at +20 to +60 mV; (iii) a transient current (I(A)) activated at membrane potentials more positive than -60 mV after a hyperpolarizing pre-pulse, but which was rapidly and totally inactivated at all potentials within its activation range; and (iv) a new K(+) current, the M current (I(M)).2. I(M) was detected as a non-inactivating current with a threshold at -60 mV. The underlying conductance G(M) showed a sigmoidal activation curve between -60 and -10 mV, with half-activation at -35 mV and a maximal value (G(M)) of 84+/-14 (S.E.M.) nS per neurone. The voltage sensitivity of G(M) could be expressed in terms of a simple Boltzmann distribution for a single multivalent gating particle.3. I(M) activated and de-activated along an exponential time course with a time constant uniquely dependent upon voltage, maximizing at approximately 150 ms at -35 mV at 22 degrees C.4. Instantaneous current-voltage (I/V) curves were approximately linear in the presence of I(M), suggesting that the M-channels do not show appreciable rectification. However, the time- and voltage-dependent opening of the M-channels induced considerable rectification in the steady-state I/V curves recorded under both voltage-clamp and current-clamp modes between -60 and -25 mV. Both time- and voltage-dependent rectification in the voltage responses to current injection over this range could be predicted from the kinetic properties of I(M).5. It is suggested that I(M) exerts a strong potential-clamping effect on the behaviour of these neurones at membrane potentials subthreshold to excitation. PMID- 6294293 TI - Changes in the plasma concentrations of free and conjugated oestrogens in heifers after treatment to induce superovulation and the relationship with number of ovulations. AB - Oestradiol-17 beta and conjugated oestrone, oestradiol-17 beta and oestradiol-17 alpha were measured in peripheral plasma of heifers treated with PMSG/PGF-2 alpha to induce superovulation. Changes in the concentrations of each hormone were synchronous, the highest level being near oestrus. For a given number of ovulations the hormone with the highest concentration was total oestradiol-17 alpha, then came total oestrone, total oestradiol-17 beta and oestradiol-17 beta. For each oestrogen, the maximum preovulatory concentration measured was significantly correlated with the number of ovulations; the regression line for total oestradiol-17 alpha was twice as steep as that for oestradiol-17 beta. It is concluded that in animals treated to induce superovulation assay of total oestradiol-17 alpha gives a better induction of the number of follicles induced to ovulate than does the more conventional assay of oestradiol-17 beta. PMID- 6294291 TI - Evidence for non-cholinergic, non-adrenergic transmission in the guinea-pig ileum. AB - To elucidate further the innervation of the longitudinal and circular muscle cells of the guinea-pig ileum, junction potentials elicited by field stimulation were recorded from both muscle layers in the presence and absence of atropine (1 2 muM) with guanethidine (10 muM) at 36 degrees C.1. In longitudinal muscle cells, a single stimulus elicited an atropine-sensitive transient depolarization (cholinergic e.j.p.), whereas in circular muscle cells, a transient hyperpolarization (non-cholinergic non-adrenergic i.j.p.) was elicited. With stimulation of nerves in the presence of atropine and guanethidine, responses of circular muscle cells to nerve stimulation were preserved while the longitudinal muscle cells generated depolarization (non-ch., non-adr. e.j.p.), hyperpolarization (non-ch., non-adr. i.j.p.) or both depolarization and subsequent hyperpolarization. These potential changes ceased with application of TTX or excess Mg(2+).2. The latency for junction potentials recorded from longitudinal muscles after field stimulation was in the following order; non-ch., non-adr. e.j.p. < cholinergic e.j.p. < non-ch., non-adr. i.j.p. The cholinergic e.j.p.s had a lower, and non-ch., non-adr. i.j.p.s a higher threshold.3. At low frequencies of stimulation (below 1 Hz) amplitudes of successively generated cholinergic e.j.p.s and non-ch., non-adr. i.j.p.s were gradually reduced, but at higher frequencies (2-20 Hz) of stimulation they were summated. The amplitudes of non-ch., non-adr. e.j.p.s were, however, not affected at low frequencies of stimulation (up to 0.5 Hz) and were summated at higher frequencies (over 1 Hz) of stimulation.4. Reversal potentials for non-ch., non-adr. e.j.p.s and i.j.p.s estimated from the amplitude of junction potentials and membrane potential were 27 and -80 mV, respectively. The reversal potential for non-ch., non-adr. e.j.p.s was higher (more negative) than that for cholinergic e.j.p.s5. Generation of i.j.p.s in longitudinal muscle cells elicited by repetitive stimulation was followed by a rebound depolarization on which was superimposed a burst of spikes. During repetitive stimulation, the amplitude of hyperpolarization was gradually reduced but the rebound depolarization and spike discharge were enhanced. Thus, while the rebound depolarization was related to the amplitude of the preceding hyperpolarization it was more related to the duration of the stimulation.6. Therefore longitudinal muscles of the guinea-pig ileum show evidence of having in addition to cholinergic and adrenergic innervation, both excitatory and inhibitory non-ch., non-adr. innervation. In the longitudinal muscle layer of the ileum, the non-ch., non-adr. excitatory fibres are more densely distributed in the terminal rather than in the proximal region, while in the case of non-ch., non-adr. inhibitory fibres, the distribution is reversed. Circular muscle cells are, however, homogenously innervated by non-ch., non-adr. inhibitory nerves. PMID- 6294294 TI - Unmasking by neuraminidase of specific chorionic gonadotrophin binding activity of human placental syncytiotrophoblast. AB - Purified human placental syncytiotrophoblast consistently failed to bind specifically to 125I-labelled hCG. Treatment of the syncytiotrophoblast with neuraminidase resulted in the ability to bind 125I-labelled hCG that was displaceable by excess of unlabelled hCG. Neuraminidase treatment removed 73.8% of the total neuraminic acid of syncytiotrophoblast. The specific binding of 125I labelled hCG increased linearly with increasing amount of neuraminidase-treated syncytiotrophoblast, was saturable and had a Ka = 1.6 x 10(7) M-1. Excess of GH, prolactin, placental lactogen or insulin did not inhibit the binding, whereas LH did so completely and FSH partly. PMID- 6294296 TI - Isotopes in the measurement of joint inflammation. PMID- 6294295 TI - Isolation of cytomegalovirus from synovial cells of a patient with rheumatoid arthritis. AB - Synovial cells from the joint of a patient with rheumatoid arthritis (RA) showed foci of cytopathic effects during their 3rd subculture in vitro. Electron microscopy of these cells revealed findings characteristic for cytomegalovirus (CMV), and immunofluorescent staining using antiserum specific for CMV was positive. The patient had classical seropositive RA, was not taking immunosuppressive agents, and had no clinical manifestations of CMV disease at the time of synovectomy 3 months earlier. Titers of antibodies to CMV were studied in the sera and synovial fluids of 3 additional patients with RA, and in 1, elevated IgM antibodies to CMV were found, indicating recent infection. Additional studies for CMV in RA are indicated. PMID- 6294297 TI - Do sulphonamide-trimethoprim combinations select less resistance to trimethoprim than the use of trimethoprim alone? PMID- 6294298 TI - Isolation and properties of metronidazole-resistant mutants of Clostridium perfringens. AB - Clostridium perfringens strains resistant to metronidazole and tinidazole were isolated from the sensitive parent strain CM288 after mutagenesis with N-methyl N'-nitro-N-nitrosoguanidine. Strain CM288 was already resistant to rifampicin and nalidixic acid; these genetic markers helped to confirm the identity of mutants. All mutants showed similar characteristics: they grew more slowly than the parent strain and failed to reach the same maximum turbidity; uptake of metronidazole and tinidazole from culture fluids was slow and end products of glucose metabolism were different from those of the parent. Pyruvate dehydrogenase activity was not detected in broken cell preparations of the mutant strains although this enzyme was readily detected in the parent strain. Changes in end products of glucose metabolism were consistent with the absence of pyruvate dehydrogenase activity because pyruvate was accumulated during growth and lactate levels were higher whereas acetate, CO2 and ethanol levels were diminished. PMID- 6294299 TI - Response of mammalian cell lines to the toxins of Escherichia coli. PMID- 6294300 TI - The ben gene of bacteriophage lambda. Mapping, identification and control of synthesis. PMID- 6294301 TI - Extrahelical bases in duplex DNA. PMID- 6294302 TI - Electron microscopic mapping of complementary sequences on single strands of bacteriophage lambda DNA. PMID- 6294303 TI - Mendelian analysis of the organization of actin sequences in Physarum polycephalum. PMID- 6294304 TI - Fine structure of the regulatory region of simian virus 40 minichromosomes revealed by DNAase I digestion. PMID- 6294305 TI - Binding sites of viral protein P19 onto Rous sarcoma virus RNA and possible controls of viral functions. PMID- 6294306 TI - Interactions of phosphate ligands with Escherichia coli aspartate carbamoyltransferase in the crystalline state. PMID- 6294307 TI - Transcription in vitro of an isolated fragment of bacteriophage T4 genome. PMID- 6294308 TI - Preliminary studies of crystals of poliovirus type I. PMID- 6294309 TI - Membrane currents related to configuration changes in the action potential of frog atrial muscle in Na- and Ca-free conditions. PMID- 6294311 TI - Isoproterenol, norepinephrine and phosphodiesterase inhibitors are blockers of the depressed fast Na+-system in ventricular muscle fibers. PMID- 6294310 TI - Role of purines in acetylcholine-induced coronary vasodilation. PMID- 6294312 TI - Mechanism of Ca2+ resistance in adult heart cells isolated with trypsin plus Ca2+. PMID- 6294313 TI - Spontaneous bladder rupture and cytomegalovirus infection complicating renal transplantation: cause or coincidence? AB - The high incidence of surgical complications following renal transplantation is well known. Urologic complications, however, present some of the most challenging problems to the transplant surgeon. The authors present here a detailed case report of spontaneous (delayed) bladder rupture (SDBR) which occurred 90 days after kidney transplantation in a recipient with cytomegalovirus infection (CMV). Urinary catheter drainage is recommended in preference to surgical intervention for the successful correction of SDBR. It is postulated further that, despite a negative bladder biopsy, CMV may have infiltrated the bladder and contributed to this "spontaneous" bladder wall rupture. PMID- 6294314 TI - Histogenetic classification of lung carcinomas. small cell carcinomas studied by light and electron microscopy. AB - Forty-six small cell carcinomas of the lung identified by light microscopy were also studied by electron microscopy in an attempt to subclassify them histogenetically. Thirty were subclassifiable into neuroendocrine secretory, mucus secretory, or epidermoid categories on the basis of their differentiated structural features; 16 were too poorly differentiated to assign to any specific cell type and were designated null. In addition to histogenetic separation, electron microscopy revealed a spectrum of average cell differentiation within each subclass except null. The widest spectrum occurred in the most frequent subclass, the neuroendocrine group. This study clearly demonstrated that the light microscopic diagnosis of small cell carcinoma does not identify a morphologically homogeneous process. Whether histogenetic separation and/or some assessment of the relative degree of differentiation have any consistent clinical value remains to be shown; however, combined light and electron microscopy should improve the precision of the initial diagnosis over that of light microscopy alone. PMID- 6294315 TI - Malignant breast lesions. PMID- 6294316 TI - Coronavirus-like particles and other agents in the faeces of children in Efate, Vanuatu. AB - Between June 1979 and July 1980, faecal specimens were collected from 100 children in Efate, Vanuatu, and examined for the presence of virus-like particles by direct negative staining electron microscopy. Of the children, 40 were hospital patients 22 of whom had gastroenteritis and 18 had other illnesses. The remaining 60 children were apparently healthy controls. Coronavirus-like particles were the most common agents encountered and were detected in approximately 20% of all three groups of children. Particles with the morphological features of calicivirus, astrovirus, adenovirus and parvovirus were also detected. PMID- 6294317 TI - Dietary bulk as a limiting factor for nutrient intake in pre-school children: IV. Effect of digestive enzymes on the viscosity of starch-based weaning foods. PMID- 6294318 TI - Changes of cellular structure and subcellular enzymatic patterns during the activation of glyconeogenesis in Tetrahymena pyriformis. PMID- 6294319 TI - [Ultrastructure of Sertoli cells in teleost fish: Abudefduf marginatus]. PMID- 6294320 TI - Factors controlling the in vitro growth pattern of human microvascular endothelial cells. AB - Monolayer cultures of endothelial cells of human dermal microvascular origin were exposed to a variety of culture conditions and in vitro differentiation of the cells assessed by light and electron microscopic examination. Restoration of a cytologic and fine structural appearance which resembled most closely that present in vivo was possible by raising the intracellular cAMP level. These cells formed junctional complexes seen in uncontracted microvessels and specialized attachment sites at their basal cell membrane, contained a complex network of bundled micro- and intermediate filaments and numerous Weibel-Palade bodies and accumulated electron-opaque deposits between the cells and the culture dish surface. PMID- 6294321 TI - Assembly of vesicular stomatitis virus: distribution of the glycoprotein on the surface of infected cells. AB - This study demonstrates that the glycoprotein of vesicular stomatitis virus clusters in the plasma membrane of infected Chinese hamster lung cells during morphogenesis and suggests that viral nucleocapsids are required for this clustering. A mutant virus (ts E-1) which is temperature sensitive for the synthesis of viral nucleocapsids but not viral membrane proteins was used. The surface distribution of the viral glycoprotein in cells infected by this virus was determined by a specific indirect immunoferritin stain. Early in infection at permissive temperatures, the glycoprotein was randomly distributed on membrane ghosts. Later, clusters of ferritin the size and shape of virus particles were seen. In contrast, ghosts prepared from virus-infected cells maintained at a restrictive temperature always had a random distribution of viral glycoprotein. PMID- 6294322 TI - Molecular biology of rotaviruses. IV. Molecular cloning of the bovine rotavirus genome. AB - A new cloning strategy has been developed for cloning the genomes of double stranded RNA viruses by using bovine rotavirus as a test system. The major modification adopted was the use of denatured polyadenylated double-stranded RNA as the template for reverse transcriptase. This allowed the two complementary strands of cDNA to be synthesized in a single reaction and removed the need for S1 nuclease digestion to remove the 5' hairpin structure normally generated in cDNA synthesis. PMID- 6294323 TI - mlt Mutants of polyoma virus. AB - New mlt deletion mutants of polyoma virus were isolated, and their abilities to produce a lytic response in mouse cells or to transform rat cells were assessed. Their properties were analyzed in terms of the sequences deleted and their effects upon the structure and functions of the viral middle and large T antigens. PMID- 6294324 TI - DNA-binding properties of a herpes simplex virus immediate early protein. AB - The herpes simplex virus alpha, or immediate early, protein ICP4 has been shown to be central to the control of the early stages of virus replication. The detailed mechanism of this control is unknown. In this communication we show that purified ICP4 was unable to bind to DNA even though the protein was capable of such activity in a crude extract. Addition of either infected- or uninfected-cell extracts to the purified protein restored its DNA-binding activity. These results suggest that ICP4 binds to DNA only via a component of uninfected cells. PMID- 6294325 TI - The B95-8 isolate of Epstein-Barr virus arose from an isolate with a standard genome. AB - Blot hybridization studies revealed that the deletion which characterizes the DNA from the B95-8 strain of Epstein-Barr virus was not present in the virus from which the B95-8 strain was derived (883L). The deletion event must have occurred during establishment of the B95-8 cell line or very soon afterward, since the deletion was present in Epstein-Barr virus DNA from a cell line established with B95-8 virus soon after it became available. The presence of the deletion correlates with decreased expression of the gp220 viral envelope glycoprotein. PMID- 6294327 TI - Further characterization of a protein kinase from foot-and-mouth disease virus. AB - Acid disruption of foot-and-mouth disease virus released a protein kinase activity that sedimented at less than 7S. This enzyme was separated into three peaks of activity by ion-exchange and hydroxylapatite chromatography. Analysis of the various enzyme fractions by polyacrylamide gel electrophoresis and silver staining revealed that one of the fractions lacked the major virion structural proteins, but still contained two or three other polypeptides. This enzyme phosphorylated mainly one protein (P17) in an in vitro assay. PMID- 6294326 TI - Detection by RNA blot hybridization of RNA sequences homologous to the BglII-N fragment of herpes simplex virus type 2 DNA. AB - RNA species, extracted at the time of peak synthesis of the alpha, beta, and gamma classes of herpes simplex virus polypeptides from lytically infected Vero cells, were examined for homology to the BglII-N fragment (map units 0.58 to 0.63) of herpes simplex virus type 2 DNA. By using northern blot analysis, two major and several minor polyadenylated RNA species showed homology to the BglII-N fragment at times corresponding to the maximum synthesis of the beta (7 h postinfection) and gamma (12 h postinfection) herpes simplex virus polypeptides. No alpha RNA homologous to the BglII-N fragment was detected. PMID- 6294328 TI - Accurate transcription of simian virus 40 chromatin in a HeLa cell extract. AB - During simian virus 40 viral maturation, a series of modifications occur which alter the composition of viral nucleoprotein complexes. As a consequence, the chromatin that is extracted from extracellular simian virus 40 virions exhibits properties that are similar to those of transcriptionally active eucaryotic chromatin. The influence of this chromatin structure on specific RNA initiation by RNA polymerase II was examined by using the in vitro HeLa cell extract of Manley et al. (Proc. Natl. Acad. Sci. U.S.A. 77:3855-3859, 1980). The 5' ends of RNA transcripts were positioned by the "run-off" assay, in which transcripts extend from the initiation site to termination sites created by restriction cleavage and by S1 nuclease analysis, using DNA probes labeled at their 5' termini. Two major early RNA transcripts, which originated at map positions 5,240 +/- 10 and 5,145 +/- 10, and two major late RNA transcripts, which originated at map positions 325 +/- 10 and 185 +/- 10, were identified. Transcripts were initiated with comparable relative efficiencies at the same 5' site when either purified DNA or chromatin was used as the template. Our results suggest that extracellular simian virus 40 virion chromatin modifications do not regulate simian virus 40 promoter selection but function to increase the accessibility of RNA promoter sequences to RNA polymerase II and allow efficient elongation of the RNA chain after the initiation event. PMID- 6294329 TI - Coronavirus proteins: biogenesis of avian infectious bronchitis virus virion proteins. AB - We examined the synthesis of viral structural proteins in cultured cells infected with the avian coronavirus infectious bronchitis virus. Tryptic peptide mapping was used to determine the structural relationships of the intracellular proteins to the virion polypeptides. Pulse-chase experiments were performed to identify precursors to the virus-specific proteins. We found that the nucleocapsid protein, P51, and the small viral membrane proteins GP31, GP28, and P23 do not undergo post-translational proteolytic processing. In contrast, GP90 and GP84, the two large virion membrane proteins, were found to be produced by cleavage of a single precursor, GP155. This demonstrated that at least one coronavirus mRNA specifies two virion proteins. PMID- 6294330 TI - Coronavirus proteins: structure and function of the oligosaccharides of the avian infectious bronchitis virus glycoproteins. AB - The recent finding that the E1 glycoproteins of murine coronaviruses contain only O-linked oligosaccharides suggested that this unusual modification might be a distinguishing feature of coronaviruses and might play an essential role in the life cycle of this family of viruses. To examine these possibilities, we analyzed the oligosaccharide moieties of the membrane proteins of the avian coronavirus infectious bronchitis virus. In addition, we determined the effect of inhibiting the glycosylation of these proteins on viral maturation and infectivity. Infectious bronchitis virus virions contain nine proteins. Four of these proteins, GP36, GP31, GP28, and P23, are closely related structurally and appear to be homologous to the E1 proteins of murine coronaviruses. We found that the oligosaccharides of GP31 and GP28 could be removed with endoglycosidase H and that neither of these glycoproteins was detectable in tunicamycin-treated cells. These two results indicated that GP31 and GP28 contain N-linked oligosaccharides. Therefore, O-linked oligosaccharides are not a universal feature of the small coronavirus membrane glycoproteins. Tunicamycin inhibited glycosylation of all of the viral glycoproteins but did not inhibit production of virions by infectious bronchitis virus-infected cells. The virions released by these cells contained only the three non-glycosylated viral proteins P51, P23, and P14. These particles were not infectious. Therefore, it appears that glycosylated infectious bronchitis virus polypeptides are not required for particle formation. However, the viral glycoproteins are apparently indispensible for viral infectivity. PMID- 6294331 TI - Genetic heterogeneity within individual bovine rotavirus isolates. AB - The genomic RNA patterns of six different bovine rotavirus isolates were analyzed on high-percentage polyacrylamide gels (12.5, 13.6, and 17.5%). In contrast to the RNA patterns exhibited by conventional gel systems, those on the high percentage gels showed an improvement in segment resolution which consequently aided in the detection of extensive band splitting in these patterns. The ability to clone out various electrophoretically distinct virus subpopulations from each of the six isolates provided an explanation for the band splitting detected by the high-resolution gels. The significance of the coexistence of genetically distinct rotavirus populations within a single host is discussed. PMID- 6294332 TI - Identification and nucleotide sequences of two similar tandem direct repeats in Epstein-Barr virus DNA. AB - Epstein-Barr virus DNA is known to have partially homologous segments, designated DL and DR, near the left and right ends of the long unique region (Raab-Traub et al., Cell 22:257-267, 1980). DL and DR are each partially composed of tandem direct repeat sequences. DL contains 11 to 14 repeats of a 124-base-pair sequence designated IR2. DR contains approximately 30 direct repeats of a 103-base-pair sequence designated IR4. The DL and DR sequences have colinear partial homology for approximately 2.4 and 1.5 kilobase pairs to the right of IR2 and IR4, respectively. IR2 and IR4 are similar sequences and evolved in part from a common ancestor. Both sequences are 84% guanine and cytosine and have limited homology to Epstein-Barr virus IR1 and to the herpes simplex virus type 1 inverted terminal repeat "a" sequence. IR2 encodes part of an abundant 2.5-kilobase persistent early EBV RNA expressed in productively infected cells, but does not encode part of the 3-kilobase Epstein-Barr virus RNA which is transcribed from the adjacent IR1-U2 region of the Epstein-Barr virus genome in latently infected cells. PMID- 6294333 TI - Non-immortalizing P3J-HR-1 Epstein-Barr virus: a deletion mutant of its transforming parent, Jijoye. AB - The P3J-HR-1 strain of Epstein-Barr virus (EBV) fails to immortalize human lymphocytes. We wished to understand the nature of the genomic alterations which correlated with the loss of this ability. As a first step, the heterogeneity of DNA molecules in the P3J-HR-1 line was eliminated by cell cloning. Then a physical map was prepared of virion DNA from one cell clone, designated FF452-3. By comparison with the genomes of two EBVs, B95-8 and FF41, which are competent to immortalize lymphocytes, we identified a total of eight modifications of BamHI and EcoRI restriction endonuclease fragments of EBV (FF452-3) DNA consisting of insertions, deletions, or loss of a restriction endonuclease recognition site. To determine which of these alterations might be responsible for the loss of transforming phenotype, we examined homologous DNA fragments of the Jijoye strain of EBV, the progenitor of the HR-1 strain which still retains the ability to immortalize lymphocytes. We also studied viral DNA in lymphocytes transformed in vitro by Jijoye virus. Six of the eight alterations were found both in Jijoye and in clonal HR-1 DNA and were presumably genomic traits characteristic of this lineage of EBV. A small deletion in the BamHI-K fragment of HR-1 DNA was not found in Jijoye virion DNA, but this deletion was present in intracellular Jijoye DNA. Thus only one major genomic lesion in HR-1 DNA, a deletion of at least 2.4 x 10(6) molecular weight of DNA from a fused BamHI-H-Y fragment, consistently distinguished Jijoye DNA from its non-immortalizing P3J-HR-1 derivative. This deletion is likely to affect EBV genes which are directly or indirectly involved in immortalizing lymphocytes. PMID- 6294334 TI - Novel forms of woodchuck hepatitis virus DNA isolated from chronically infected woodchuck liver nuclei. AB - We cloned several unique forms of woodchuck hepatitis virus, a DNA virus closely related to hepatitis B virus, from a chronically infected woodchuck liver. Each of the three clones contained more than two genome equivalents of viral sequences with extensive rearrangements and no detectable cellular sequences. From the frequency by which they were isolated from a library of recombinant clones, we estimate that they are present in approximately one copy per cell. Of a total of 11 sites at which rearrangements were mapped in the clones, 10 occurred between segments of opposite polarity, and 1 occurred between segments of the same polarity. The possible significance of these findings to the persistence of virus production in infected cells is discussed. PMID- 6294335 TI - Separation and properties of two kinds of simian virus 40 late transcription complexes. AB - Simian virus 40 (SV40) transcription complexes were labeled in cells with 3-min pulses of [(3)H]uridine 48 h after infection and were extracted from nuclei in isotonic buffer or in a buffer containing Sarkosyl. In sucrose gradients, the labeled complexes sedimented faster than both free RNA and most SV40 nucleoproteins. Most of the pulse-labeled nascent RNA hybridized to the entire late region of SV40, remained bound to viral DNA in Cs(2)SO(4) gradients, and ranged in size from a few nucleotides to about 5,000 nucleotides, with a peak at about 700. In contrast, the SV40-associated RNA polymerase activity in the same preparations sedimented near the major peak of SV40 nucleoproteins and was clearly separated from the transcription complexes bearing pulse-labeled nascent RNA. The two kinds of transcription complexes were released from isolated nuclei at different rates. Complexes bearing pulse-labeled RNA were released immediately when the nuclei were agitated in a Dounce homogenizer in isotonic buffer, whereas most of the complexes bearing RNA polymerase active in vitro were released more slowly, during subsequent incubation of the nuclei at 0 degrees C. Since the complexes bearing pulse-labeled nascent RNA were virtually inactive in vitro, the blocked complexes described by Laub et al. (Proc. Natl. Acad. Sci. U.S.A. 77:3297 3301, 1980) probably account for almost all the SV40-associated RNA polymerase activity studied previously by many investigators. New procedures must be developed to preserve the activity of the pulse-labeled complexes if the many advantages of the SV40 system for studying transcription by nucleoprotein complexes in vitro are to be realized fully. PMID- 6294336 TI - Development of a mouse mammary tumor virus-negative mouse strain: a new system for the study of mammary carcinogenesis. AB - All inbred strains of mice transmit one or more copies of mouse mammary tumor virus (MMTV) DNA integrated as proviral sequences. This complicates efforts to define viral-induced mammary carcinogenesis. Here we report the use of surgical nonlethal splenectomy in tissue typing mice and the development of an MMTV negative mouse strain. The MMTV-negative strain allows study of the involvement of non-MMTV genes in mammary carcinogenesis. In addition, it can be used as a sterile background into which MMTV variants can be introduced. Through the techniques described here, mice containing single MMTV loci or specific combinations can be specially chosen and rapidly developed. In this manner, the oncogenecity of particular MMTV variants may be assessed. PMID- 6294337 TI - Identification and characterization of a DNase induced by Epstein-Barr virus. AB - The diterpene ester promoter of mouse skin tumors, 12-O-tetradecanoyl-phorbol-13 acetate, induced a DNase activity in the Epstein-Barr virus-producer cell line P3HR-1. The elution patterns of the enzyme from DEAE-cellulose, phosphocellulose, and DNA-cellulose columns were different from virus-associated DNA polymerase activity. The partially purified activity could be neutralized to the extent of 90% by sera of patients with nasopharyngeal carcinoma. Purified immunoglobulin G from sera of nasopharyngeal carcinoma patients inhibited this enzyme and that obtained from superinfected Raji cells to the same extent. The partially purified enzyme preferred native DNA as a substrate over denatured DNA and 3'-terminally labeled activated calf thymus DNA. The activity was inhibited by high ionic strength. Phosphonoformic acid did not have any effect on this enzyme activity. PMID- 6294338 TI - In vitro synthesis and assembly of picornaviral capsid intermediate structures. AB - Cell-free translation of encephalomyocarditis RNA in extracts of rabbit reticulocytes results in the synthesis of viral proteins indistinguishable from those produced during virus infection of cells. The viral capsid proteins are produced in an active form capable of assembly into viral capsid intermediate structures. Protomers (5S), pentamers (14S), and shell-like structures (75 to 85S) can be detected after prolonged incubation in the extracts. Proteolytic cleavage of capsid precursor proteins appears to be a prerequisite for assembly, in apparent contrast to cell-associated assembly. Assembly of pentamers is also preceded by conversion of protein epsilon 1 to epsilon in a step which may reflect an amino-terminal blocking reaction. PMID- 6294339 TI - Molecular cloning of biologically active proviral DNA of the anemia-inducing strain of spleen focus-forming virus. AB - Previously, we have molecularly cloned proviral DNA of a polycythemia-inducing strain of the spleen focus-forming virus (SFFVp). In this paper, we report that unintegrated proviral DNA of the anemia-inducing strain of SFFV (SFFVA) has been molecularly cloned into pBR322. This molecularly cloned DNA retains the biological activity of SFFVA, as infectious SFFV can be recovered from the DNA clone by marker rescue using a previously described two-stage cotransfection assay (Linemeyer et al., J. Virol. 35:710-721, 1980). The recovered SFFV retains an important property of the initial SFFVA which distinguishes SFFVA from SFFVP, namely, the ability of SFFVA to cause proliferation of erythroid cells in which hemoglobin synthesis is erythropoietin dependent. By utilizing a marker rescue technique, the splenomegaly and anemia characteristic of SFFVA-induced disease have been traced to a DNA fragment of SFFVA containing sequences coding for the env gene product. gp52. The results suggest that the differences in pathogenicity between SFFVP disease and SFFVA disease are an intrinsic property of the env gene products of these two variants of Friend virus, and future studies with the molecular clones of each strain should allow us to map regions of each env gene responsible for common and distinctive features of the erythroproliferative diseases induced by each virus. PMID- 6294340 TI - Translational control of vesicular stomatitis virus protein synthesis: isolation of an mRNA-sequestering particle. AB - An mRNA-ribonucleoprotein particle (mRNP) was found in vesicular stomatitis virus (VSV)-infected Chinese hamster ovary cells. The particle was present 3 and 4.5 h after infection but was barely discernible at 2 h. The mRNP (buoyant density, 1.56 g/cm3), which cosedimented with viral nucleocapsid in a sucrose density gradient at approximately 120 to 160S, was separable from nucleocapsid (buoyant density, 1.31 g/cm3) by CsCl density gradient centrifugation. It contained all five VSV mRNAs and, almost exclusively, viral N protein. Some host mRNA and host protein was also present in the particle. The intact mRNP was incapable of stimulating protein synthesis in an in vitro protein-synthesizing system, although the VSV mRNA isolated from the particle by phenol extraction was functional in vitro. In contrast, intact polysomes stimulated cell-free protein synthesis to the same extent as purified polysomal mRNA. By 4.5 h after infection, 97% of the functional mRNA in vivo was associated with the mRNP, and only 3% was on polysomes. The amount of polysomal mRNA at 4.5 h after infection was only 31% of that found at 2 h after infection; this was reflected by the 76% decrease observed in the rate of in vivo protein synthesis at 4.5 h relative to that found at 2 h. Thus, it appears that the mRNP serves as an organelle which sequesters the large excess of VSV mRNA that is normally made during secondary transcription. PMID- 6294342 TI - A locus that enhances the induction of endogenous ecotropic murine leukemia viruses is distinct from genome-length ecotropic proviruses. AB - The segregation of genes that enhance the induction of ecotropic murine leukemia viruses (In loci) has been compared with the segregation of ecotropic-specific nucleotide sequences in 12 low-leukemic mouse strains and 18 recombinant inbred strains. Endogenous ecotropic viruses of these strains are of genome length and structurally similar to AKR ecotropic proviruses. Low-leukemic strains of related pedigree contain ecotropic proviruses at common integration sites. Loci previously identified which enhance induction of ecotropic viruses (In genes) were correlated with the inheritance of ecotropic viral sequences in inbred low leukemic mouse strains and in CXB recombinant inbred mouse strains. However, four BXH recombinant inbred strains were observed to possess an In gene(s) yet lack the probed envelope gene region for the corresponding endogenous ecotropic virus. These observations indicate that at least one gene that enhances ecotropic virus expression in vitro is encoded by DNA sequences outside ecotropic proviruses or by subgenomic viral sequences. PMID- 6294343 TI - Alteration in the simian virus 40 maturation pathway after butyrate-induced hyperacetylation of histones. AB - The role of histone acetylation in the replication and maturation pathways of simian virus 40 was assessed. Histones were hyperacetylated by briefly exposing infected cells to sodium butyrate. Viral DNA in cells exposed to butyrate was found to reenter replication to a greater extent and mature to the previrion form to a lesser extent than viral DNA in control cells. Previrions formed in the presence of butyrate had altered sedimentation properties. These data suggest that increased acetylation of histones is not the signal for removal of DNA from the pool of molecules available for replication. It appears, in fact, that hyperacetylation retards entry into and progression along the maturation pathway. PMID- 6294341 TI - Structural features of the herpes simplex virus alpha gene 4, 0, and 27 promoter regulatory sequences which confer alpha regulation on chimeric thymidine kinase genes. AB - Previous studies have shown that herpes simplex virus genes form three groups, alpha, beta, and gamma, whose expression is coordinately regulated and sequentially ordered in a cascade fashion. Chimeric genes constructed by fusion of the coding and 5' nontranslated leader sequences of the thymidine kinase (TK) gene to the sequences upstream from the site of initiation of transcription of alpha genes 4 and 27 are regulated as alpha genes and are induced in cells converted to TK+ phenotype by infection with TK- virus. In alpha gene 4 (S. Mackem and B. Roizman, Proc. Natl. Acad. Sci. U.S.A. 79:4917-4921, 1982), both the promoter and the regulatory region are separable and movable. The promoter permits expression but not induction when fused to TK in the noncoding leader region of the gene. The regulator, when fused to the promoter of an expressible but noninducible portion of the natural beta TK, renders the gene inducible as an alpha gene; it consists of multiple regulatory units acting cumulatively. In this paper, we report on the precise site of initiation of transcription of alpha gene 0 within the inverted b sequences of the L component of viral DNA. We also report the following. (i) The chimeric gene consisting of the coding and 5' nontranslated leader regions of the TK gene fused to portions of the domain of alpha gene 0 extending largely upstream from the site of initiation of transcription of alpha gene 0 was regulated in the same fashion as the alpha 4- and alpha 27-TK chimeras. The regulatory region in the alpha gene 0 is largely upstream from nucleotide - 140. (ii) The promoter-regulatory regions of alpha genes 0, 4, and 27 share TATA sequences, A + T-rich (consensus) sequences occurring in regulating regions of alpha genes 0 and 4 in more than one copy, and multiple G + C-rich inverted repeats. The relation of these sequences to the function of the promoter-regulatory regions of the alpha genes is discussed. PMID- 6294344 TI - Structure and synthesis of a simian virus 40 super T-antigen. AB - Mouse cells transformed by simian virus 40 often contain virus-coded tumor antigens distinct from those synthesized in productively infected permissive cells. The SV3T3 C120 cell line produces no large T-antigen of apparent molecular weight 94,000 but instead a super T-antigen of apparent molecular weight 145,000. We used recombinant DNA techniques to isolate the template for this super T antigen and determined its structure by DNA sequencing. The integrated viral early transcription unit contains an in-phase, perfect tandem duplication of 1,212 base pairs. Transfer hybridization and endonuclease S1 mapping experiments were performed to elucidate the structures of the stable, cytoplasmic mRNAs of SV3T3 C120 cells, mRNAs of 3.9 and 3.6 kilobases, containing the small t- and large T-antigen splices, respectively, were transcribed from the internally duplicated early transcription unit. We showed by in vitro translation that these mRNAs encode small t-antigen and the super T-antigen of molecular weight 145,000. Peptide mapping studies of the SV3T3 C120 super T-antigen were consistent with its being derived from an internally duplicated template, since the protein has methionine and cysteine tryptic fingerprints virtually identical to those of normal large T-antigen, with certain methionine peptides present in greater than one molar yield. PMID- 6294345 TI - Functional analysis of a simian virus 40 super T-antigen. AB - The SV3T3 C120 line of simian virus 40-transformed mouse cells synthesizes no large T-antigen of molecular weight 94,000 but instead a super T-antigen of molecular weight 145,000. In the accompanying paper (Lovett et al., J. Virol. 44:963-973, 1982), we showed that the integrated viral DNA segment SV3T3-20-K contains a perfect, in-phase, tandem duplication of 1.212 kilobases within the large T-antigen coding sequences. Our data suggested that this integrated template encodes mRNAs of 3.9 and 3.6 kilobases, the smaller of which directs the synthesis of the super T-antigen of molecular weight 145,000. We transfected the DNA segment SV3T3-20-K into nonpermissive rat cells and into TK- mouse L cells and analyzed the T-antigens and viral mRNAs in the transfectants; these data prove directly the coding assignments suggested previously. The super T-antigen retained the ability to induce morphological transformation, and may even transform better than the wild-type protein. It also retained the ability to bind to the cell-coded p53 protein. Transfection into permissive CV-1 cells showed that the super T-antigen encoded by SV3T3-20-K was incapable of initiating DNA replication at the viral origin. The duplication in SV3T3-20-K thus defines a mutation which separates the transformation and DNA replication functions of large T-antigen. We discuss why such mutations may be selected in transformed cells. PMID- 6294346 TI - Association of viral particles and viral proteins with membranes in SA11-infected cells. AB - Electron microscopy after negative staining of SA11-infected cell homogenates revealed that most of the viral particles are associated with membrane-like material. Many of the particles seemed to be fully enveloped in a membrane. This association could also be detected by the observed cosedimentation of viral proteins and cell membranes. Pulse-chase experiments showed that viral glycoproteins rapidly associate with membranes, whereas most of the structural proteins appearing in the soluble fraction immediately after the pulse were slowly chased into the membrane fraction. The membranes could be further fractionated into at least four fractions differing in density and containing a different distribution of viral proteins. Also, the distribution of label into each of these membrane fractions changed after long chase periods. The inhibition of glycosylation with tunicamycin yielded viral particles without an outer layer, but did not affect the described association with membranes. The possible relationship of this finding to the maturation of the virion is discussed. PMID- 6294348 TI - Primary signet ring cell adenocarcinoma of the bladder (linitis plastica of the bladder): report of a case and review of the literature. AB - Signet ring cell adenocarcinoma of the bladder is a rare tumor that usually presents as a diffuse and infiltrating neoplasm, similar to the linitis plastica type of gastric adenocarcinoma. We report the thirteenth case of primary signet ring cell adenocarcinoma of the bladder, which is documented by light microscopic, immunohistochemical and electron microscopic studies. the pertinent medical literature is reviewed and analyzed. In general, this tumor present with hematuria and irritative symptoms, and may yield equivocal radiologic and cystoscopic findings. It is characterized by a rather rapid growth rate, infiltration of the surrounding pelvic organs, and variable response to radiotherapy and radical surgery. The prognosis is apparently favorably modified by early detection of localized forms of the tumor and aggressive treatment. PMID- 6294347 TI - Lytic action of cloned phi X174 gene E. AB - The phi X174 lysis gene E was placed under control of the lac promoter by cloning into the multicopy plasmid pBH20. Other phi X174 gene sequences were removed by nuclease digestion. Expression of gene E was shown to be necessary and sufficient to produce lysis phenomena exhibited by infection with intact phage. Lysis, its inhibition by MgSO4 and spermine, its progression through a spheroplasting stage, and its dependence on an early chloramphenicol-sensitive step were reproduced in clones induced for expression of the E gene product. Escherichia coli clones carrying the E gene not under lac control, and clones under lac control but only minimally induced for gene E expression, exhibited morphological aberrations consistent with the view that the mechanism by which gene E mediates cell lysis is related to host cell division processes. PMID- 6294349 TI - Hepatoma associated with diethylstilbestrol therapy for prostatic carcinoma. PMID- 6294350 TI - The effect of the chronic administration of a potent luteinizing hormone releasing hormone analog on the rat prostate. AB - In order to assess the effect of the chronic administration of a potent luteinizing hormone releasing hormone analog, (D-SER(But)6) LHRH (1-9) nonapeptide-ethylamide (Buserelin, HOE 766) on the pituitary gonadal axis, and the prostate, adult male Wistar rats were administered either 0, 3, 10 or 50 micrograms./kg. body weight Buserelin subcutaneously daily. At 7, 21, 35 and 42 days of treatment, groups of animals were sacrificed and certain serum endocrine and grave metric parameters determined. In addition, at 1, 21 and 42 days of treatment the 1-hour response of serum LH and serum testosterone to a single injection of 10 micrograms./kg. body weight Buserelin was determined. All treatment doses had similar effects. Serum prolactin and the basal and "acute" response of serum LH to Buserelin (+ delta 5,000 per cent) were unaltered throughout treatment. Testes weight, testicular LH receptor content, and basal and "acute" concentrations of serum testosterone were markedly decreased by 42 days of treatment (48, 89, 88 and 88 per cent, respectively). Although seminal vesicle weight declined 50 per cent at 42 days of treatment, prostate weight was not altered from initial weight, but was significantly lower than age matched control at 42 days of treatment. Buserelin remains a potent stimulator of pituitary LH release even during chronic administration. It markedly reduces serum testosterone through a predominant testicular site of action. Buserelin treatment inhibits the growth of the normal prostate, but does not cause its regression. PMID- 6294351 TI - Interstitial nephritis after cimetidine but not ranitidine. PMID- 6294352 TI - Cannabinoids in plasma after passive inhalation of marijuana smoke. PMID- 6294353 TI - Tumor lysis syndrome after induction chemotherapy of small-cell bronchogenic carcinoma. PMID- 6294354 TI - [A study of the disc sensitivity test for cefotaxime]. AB - Susceptibility of 101 strains of 29 bacterial species to cefotaxime were determined by the 2-fold agar dilution method in parallel with the diameter of inhibition zone by the single-disc method, under the experimental condition established by Kanazawa. The experiments demonstrated significant correlation between MIC by the dilution method and diameter of inhibition zone in each of conventional assay of the over-night (about 16 hours) incubation, delayed assay (about 24 hours incubation), and rapid assay (after 3 approximately 4 or 5 approximately 6 hours incubation), thus confirming applicability of the single disc assay for cefotaxime. Analysis of the data obtained by using cefotaxime disc containing 30 micrograms revealed the primary regression equation to be: D (diameter, mm) = 26.5-9.4 log MIC (microgram/ml) in conventional assay, D = 33.1 11.7 log MIC (microgram/ml) in delayed assay, D = 16.8 log MIC (micrograms/ml) in 3-4 hours rapid assay and D = 21.4-6.6 log MIC (micrograms/ml) in 5-6 hours rapid assay, respectively. The range of variations in MICs estimated from the diameter of inhibition zone by the disc test was then calculated in comparison with that in MIC determined by the 2-fold agar dilution assays, as reference for the experimental errors which may be involved in the estimation of MIC of cefotaxime by the single-disc assay. PMID- 6294355 TI - [Fundamental and clinical investigations of cefotaxime in neonates]. AB - Fundamental and clinical investigations of cefotaxime were carried out in neonates. The following results were obtained. 1. Seven neonates with serious infections caused by identifiable pathogens, including Group B streptococcal meningitis and Group A streptococcal sepsis, were treated by intravenous bolus injection of 20-200 mg/kg of cefotaxime 2 or 3 times daily (60-400 mg/kg/day). The clinical efficacy of cefotaxime was assessed to be good in 6 patients and fair in 1 patient. Bacteriological efficacy was evaluable in 4 patients, all of whom displayed complete eradication of pathogens. 2. Among 22 neonates administered cefotaxime, adverse reactions appeared in 3 patients. Adverse reactions consisted of a transient skin rash in 1 patient and elevation of GOT in 2 patients. 3. Serum concentrations of cefotaxime and desacetyl cefotaxime were investigated in 8 mature infants and 5 immature infants on days 0-7 postpartum. A single intravenous injection of 20 mg/kg produced peak serum concentrations of 31.8-49.7 mcg/ml, associated with a half-life of 1.38-4.47 hours, in mature infants and peak serum concentrations of 35.5-55.0 mcg/ml, associated with a half life of 3.22-6.43 hours, in immature infants. On days 0-2 postpartum the half life was longer than on subsequent days. This tendency was particularly remarkable in immature infants. Serum concentrations of desacetyl cefotaxime displayed high individual variations; no consistent trends were noted. 4. Cefotaxime and desacetyl cefotaxime serum concentrations were studied in 3 neonates undergoing exchanged transfusion (exchanged volume 177-180 ml/kg) on 1-4 days postpartum. Serum concentrations of cefotaxime after exchanged transfusion were equivalent to 32.6-63.9% of the pretransfusion level, while those of desacetyl cefotaxime were 75.2-106% of the pretransfusion level. 5. Minimal inhibitory concentration (MICs) and minimal bactericidal concentration (MBCs) of cefotaxime were determined against clinical isolates. MICs for inoculum sizes of 10(8)/ml and 10(6)/ml were respectively 3.13-25 mcg/ml and 3.13-25 mcg/ml against S. aureus, 0.024 mcg/ml and 0.012 mcg/ml against Group A Streptococcus, 0.05 mcg/ml and 0.05 mcg/ml against Group B Streptococcus and 0.39 mcg/ml and 0.1 mcg/ml against E. coli. MBCs for an inoculum size of 10(6)/ml were 3.13-100 mcg/ml or over against S. aureus, 0.012 mcg/ml against Group A Streptococcus, 0.39 mcg/ml against Group B Streptococcus and 1.56 mcg/ml against E. coli. PMID- 6294356 TI - [Fundamental and clinical studies of cefotaxime in neonates and immature infants]. AB - Cefotaxime (CTX) was used in the treatment and prophylaxis of infections in neonates and immature infants. The following results were obtained. 1. Mean serum concentrations (bioassay) 30 minutes after a single intravenous injection of about 20 mg/kg of CTX were 44.5 mcg/ml in neonates and 47.2 mcg/ml in immature infants aged 0-3 days, 45.8 mcg/ml in neonates and 56.4 mcg/ml in an immature infant aged 4-7 days and 40.6 mcg/ml in neonates and 38.1 mcg/ml in immature infants aged 8 or more days. Six hour values were respectively 10.9 mcg/ml, 17.0 mcg/ml, 4.6 mcg/ml, 13.4 mcg/ml, 3.8 mcg/ml and 2.7 mcg/ml. 2. Mean serum concentration half-lives were 3.0 hours in neonates and 3.2 hours in immature infants aged 0-3 days, 1.8 hours in neonates and 3.2 hours in an immature infant aged 4-7 days, and 1.5 hours in neonates and 1.6 hours in immature infants aged 8 or more days. 3. Urinary recovery rates were 0.8-78.0% for 0-6 hours after treatment. 4. Adequate clinical efficacy can be expected by the intravenous injection of CTX in doses of 20 mg/kg 2 times daily, every 12 hours, in neonates and immature infants aged 0-3 days, 20 mg/kg 3 times daily, every 8 hours, in neonates and immature infants aged 4-7 days, and 20 mg/kg 3 to 4 times daily, every 6-8 hours, in neonates and immature infants aged 8 or more days. 5. The clinical efficacy of CTX was good in all 4 cases of sepsis (including suspected case), excellent in 1 case of urinary tract infection, and good in all 4 cases of fever of unknown origin for a cure rate of 100%. 6. Adverse reactions were not noted in any cases. PMID- 6294357 TI - [Study of intravenous cefotaxime therapy in neonates]. AB - It has been proven that, when cephem group antibiotics are administered intravenously to neonates, the peak serum level of the drug is higher, and the half-life is longer, compared with the values attained in suckling infants. The same pattern is seen even with cefmetazole. This is, the Post- and Perinatal Period Research Group recently reported that, when 20 mg/kg of cefmetazole was administered intravenously, the mean serum half-life of the drug was 4.18 hours in infants up to 3 days after birth, while by about the age of 2 weeks there was no longer a difference with suckling infants. In the present study, cefotaxime was administered at a dosage level of 20 mg/kg by intravenous drip infusion over a 30-minute period. The administered subjects were a 16-day-old neonate and a 45 day-old suckling infant, and the serum level of cefotaxime was monitored. The peak concentration was found to be higher in the younger subject, and the half life in the serum was longer, i.e., 2.52 hours compared with 1.5 hours in the suckling infant. In addition, 4 cases of newborn infection were treated with cefotaxime at 120--504 mg/day (approximately 35--300 mg/kg/day), given intravenously for a period of 6 to 21 days. Clear clinical efficacy and bacteriological efficacy were achieved in relation to 1 case of staphylococcal pneumonia, 1 case of septicemia compounded by purulent meningitis caused by Enterobacter aerogenes and 1 case of fever of undetermined origin. The following summarizes the results of the present study. 1) There was no adverse effect exerted on the hepatic or renal functions. 2) Cefotaxime was efficacious in the treatment of neonates suffering from infections caused by staphylococci and a Gram-negative rod. PMID- 6294358 TI - [Basic and clinical studies on the use of cefotaxime in neonates]. PMID- 6294360 TI - [Studies on the pharmacokinetics of cefotaxime in neonates]. AB - The pharmacokinetics of cefotaxime was investigated in neonates. The following results were obtained. 1. The peak serum concentration of cefotaxime, seen 15 minutes after a single intravenous of 20 mg/kg, was 51.6 +/- 9.3 mcg/ml by bioassay. After 6 hours the mean serum concentration decreased to 5.6 +/- 3.1 mcg/ml. Concentrations obtained by HPLC paralleled those determined by bioassay. The peak serum concentration of the desacetyl metabolite was attained 30 minutes to 2 hours after injection. The mean serum desacetyl metabolite concentration was about 1/2.5 times higher than the cefotaxime concentration. The half-life was inversely related to the age of the neonates, decreasing to 1.64 hours on day 11 postpartum. 2. Serum concentrations determined by bioassay and HPLC after administering a dose of 10 mg/kg of cefotaxime by 30-minute intravenous drip infusion were comparable. The peak serum concentration at the completion of intravenous drip infusion was 21.0 mcg/ml by bioassay. The half-life of cefotaxime was 2.85 hours. The peak serum concentration of the desacetyl metabolite, seen at the completion of intravenous drip infusion, was about 1/2 times that of the peak cefotaxime concentration. 3. The peak serum concentration at the completion of a 30-minute intravenous drip infusion of 20 mg/kg displayed a mean value of 33.5 +/- 10.3 mcg/ml by bioassay. After 6 hours the mean serum concentration was 4.0 +/- 1.0 mcg/ml. The peak serum concentration of the desacetyl metabolite, seen at the completion of infusion to 2 hours thereafter, was equivalent to about 1/2.2 the peak cefotaxime concentration. 4. Mean urinary excretion rate of cefotaxime in 2-day-old neonates was 23.4% by bioassay 6 hours after a 30-minute intravenous drip infusion of 10 mg/kg. The mean urinary excretion rate of the desacetyl metabolite was 8.7%. Mean 6-hour excretion rates in 2-day-old and 4-day-old neonates administered 20 mg/kg of cefotaxime by 30 minute intravenous drip infusion were 6.2% and mean 37.7%, respectively. The corresponding values for the desacetyl metabolite were 2.4% and 12.4%, respectively. PMID- 6294359 TI - [Study on the use of cefotaxime in neonates]. PMID- 6294361 TI - [Evaluation of cefotaxime in the treatment of infections in the newborn]. PMID- 6294362 TI - [Clinical results and pharmacokinetics of cefotaxime in newborn infants]. AB - One full-term newborn infant and 2 premature ones were treated with cefotaxime for the treatment of suspected sepsis and umbilical suppurative inflammation. Pathogenic organisms could not be identified in all cases. A good result was obtained with the case of suspected sepsis. But the other 2 cases were not evaluable because underlying diseases such as massive pulmonary atelectasis or respiratory distress syndrome masked the effects of this agent. Serum levels of cefotaxime in 3 of the 4 cases were determined with bioassay. Time courses of the serum levels in 2 of them resulted in peculiar biphasic disappearance curves. This fact implies the possibility that desacetylation of cefroxime proceeds also in newborns as in adults and that desacetyl metabolite accumulates in the body owing to the premature function of the neonatal kidney. PMID- 6294363 TI - [Pharmacokinetic, bacteriological and clinical studies of cefotaxime in newborn and immature infants]. PMID- 6294364 TI - [Clinical studies of cefotaxime in perinatal infection]. AB - The therapeutic efficiency of a new cephalosporin derivative, cefotaxime, which is stable against beta-lactamase hydrolysis, has been studied in cases involving perinatal infection. The following results have been obtained. 1. In the treatment of 7 cases of infection, the preparation showed excellent efficacy in 1 case and good efficacy in 5 cases except 1 unknown case. 2. This drug has demonstrated its efficacy in the treatment of 5 cases of infections refractory to ABPC, out of which 1 had excellent and 3 had good results. 3. No side effects were observed in any of our patients. In conclusion, this drug shows excellent efficacy and high safety in the treatment of cases involving infection. PMID- 6294365 TI - [Studies on the transplacental passage of cefotaxime in late pregnancy]. AB - The transplacental passage of a single intravenous dose of cefotaxime (CTX), 1,000 mg, was examined in 11 pregnant women undergoing delivery at term by cesarean section. The results were as follows. 1. Measurements by HPLC: After a single 1,000 mg intravenous dose, the maternal blood level of CTX took the following course: 40.1 +/- 3.4 mcg/ml (mean +/- S.E.) at 15 minutes, 22.5 +/- 1.9 mcg/ml at 30 minutes, 10.8 +/- 0.9 mcg/ml at 60 minutes and 3.2 +/- 0.4 mcg/ml at 120 minutes. Slightly high maternal blood levels of 0.7 and 1.1 mcg/ml were obtained at child deliveries made at 200 minutes, but only a trace amount was found at a child delivery at 356 minutes. The CTX level in the umbilical cord blood was comparatively high until the measurement at 222 minutes, and then declined. IN the amniotic fluid, peak levels were observed at 222 to 252 minutes, thereafter decreasing slowly. Desacetyl-CTX, a metabolite of CTX, showed maternal blood levels of 5.8 +/- 0.5 mcg/ml at 15 minutes, 6.3 +/- 0.6 mcg/ml at 30 minutes, 6.8 +/- 0.7 mcg/ml at 60 minutes and 4.9 +/- 0.7 mcg/ml at 120 minutes. Thus, the maternal blood level of desacetyl-CTX showed an increasing tendency until 60 minutes after intravenous administration, and decreased rapidly from 120 minutes onwards. No consistent tendency was noted in desacetyl-CTX levels in the umbilical cord blood and the amniotic fluid. 2. Measurements by bioassay: Maternal blood levels of CTX determined by bioassay were 45.2 +/- 3.3 mcg/ml at 15 minutes, 25.8 +/- 2.2 mcg/ml at 30 minutes, 12.4 +/- 1.0 mcg/ml at 50 minutes adn 4.0 +/- 0.5 mcg/ml at 120 minutes. At the time of child delivery, maternal blood levels of CTX were 1.1 and 2.6 mcg/ml at 200 minutes, and then decreased slowly. Trace levels were noted at 336 minutes and CTX was undetectable at 356 minutes. 3. The HPLC study demonstrated that the level of CTx remained comparatively high in the umbilical cord blood and the amniotic fluid even 6 hours after administration (0.3--0.4 mcg/ml and 4.0 mcg/ml, respectively). This result suggests, in consideration of CTX's MIC values for causative organisms, that CTX will produce a sufficient clinical effect in perinatal infections. No side effects were observed in the mothers or their babies. PMID- 6294366 TI - [A study of clinical application of cefotaxime in the perinatal period]. AB - Pharmacokinetic studies of cefotaxime (CTX) were carried out in perinatal mothers and infants. CTX was promptly absorbed after intravenous injection, intravenous drip infusion and intramuscular injection in pregnant women, producing dose related peak blood levels. Placental passage to the fetus was favorable. After intravenous injection, intravenous drip infusion and intramuscular injection of 500--1,000 mg of CTX, drug concentrations of the cord blood, amniotic fluid and fetal blood exceeded MICs of the main pathogenic organisms. By administration of this dose 1 to 2 times daily it is possible to successfully prevent or treat uterine infections. Passage of CTX into the milk of lactating mothers was minimal, suggesting that only minute quantities can possibly be transferred from the milk to newborn infants. Absorption of CTX in neonatal infants was prompt. The peak blood levels of 47.9 micrograms/ml were attained 15--30 minutes after intravenous injection of 20 mg/kg. The half-life ranged from 2.4--5.56 hours, depending on the number of days postpartus. CTX was effective in the prophylaxis and therapy of perinatal uterine infections. PMID- 6294367 TI - [Fundamental and clinical studies on cefotaxime in the perinatal period]. AB - Fundamental and clinical studies on the perinatal use of cefotaxime (CTX, HR 756), a new cephalosporin antibiotic, were done, with the following results. 1. Following intravenous administration of 2 g of CTX, transport of CTX from maternal serum to umbilical cord serum and to amniotic fluid was found to be good. CTX was not transferred into mother's milk. 2. In clinical use, CTX was given to 16 pregnant patients with premature rupture of the membrane. It showed excellent efficacy in preventing perinatal infection. Five patients with perinatal infections were administered CTX, and in 4 patients CTX had good efficacy. No side effects were noted in any cases. PMID- 6294369 TI - [Study of cefotaxime in the perinatal period]. PMID- 6294368 TI - [Fundamental and clinical studies of cefotaxime during the perinatal period in pregnant women]. AB - Fundamental and clinical studies of cefotaxime (CTX) were carried out in pregnant women. The following results were obtained. CTX was more rapidly eliminated from the serum of pregnant women than that of adult males. Urinary excretion of CTX in pregnant women was comparable to that in nonpregnant women and adult males. Passage of CTX to the embryo, fetus and fetal appendages was minimal. Peak amniotic fluid concentration (9.8 mcg/ml) was attained at 3.5 hours after administration of CTX and gradually declined thereafter. This amniotic fluid concentration was sufficiently higher than reported MIC90 of CTX against E. coli strains. CTX was used in the treatment of 6 pregnant patients with acute pyelonephritis and 2 with puerperal infections. The bacteriological and clinical responses were both 100%. Since passage of CX into the amniotic fluid is favorable, CTX can be expected to be effective for the prophylaxis of intrauterine amniotic infection associated with early rupture of the membrane. CTX was used in the treatment of a neonate with purulent meningitis. The clinical response was effective. CTX did not cause any noteworthy adverse reactions or laboratory data abnormalities in our patients or neonates. PMID- 6294370 TI - Reduction of experimentally produced acute myocardial infarction size by a new synthetic inhibitor, NCO-700, against calcium-activated neutral protease. AB - Calcium-activated neutral protease (CANP) might be involved in the irreversible degradation of myocardial proteins in the ischemic region, leading to the loss of contractility. The new compound, NCO-700, and its analogues were synthetized against CANP. Among these analogues, NCO-700 was the most potent to reduce the size of acute myocardial infarction, which was produced by coronary artery ligation in rabbits, in vivo, although it showed less powerful action to inhibit CANP activity in vitro. The new reagent, NCO-700 might be promising to reduce acute myocardial infarction size and beneficial for the clinical studies, because it had no action to reduce cardiac muscle contractility, compared with beta antagonist or calcium-channel blockades. PMID- 6294371 TI - [An adult case of acute erythroblastopenia]. PMID- 6294372 TI - [Function and structure of the neuromuscular junction]. PMID- 6294373 TI - [Polyneuropathy: hereditary neuropathy]. PMID- 6294374 TI - [Polyneuropathy: toxic neuropathy]. PMID- 6294375 TI - [Symptoms often seen in peripheral nerve and muscular diseases]. PMID- 6294377 TI - [Nerve biopsy]. PMID- 6294376 TI - [Application of electrophysiologic tests to the diagnosis of peripheral nerve and muscular diseases]. PMID- 6294378 TI - [Ectopic hormones]. PMID- 6294379 TI - [Basic fetoprotein (BFP)]. PMID- 6294380 TI - [Histological classification of gastric carcinoma]. PMID- 6294381 TI - [Hepatitis-associated antigen and antibody in patients with hepatocellular carcinoma]. PMID- 6294382 TI - [2 cases of gastric cancer with marked calcification]. PMID- 6294383 TI - The effects of silica on lipid peroxidation, and the production of superoxide radicals by phagocytizing rabbit macrophages. PMID- 6294384 TI - The effect of gentamicin and mercuric chloride on cyclic AMP and lipoperoxide in rat kidneys. PMID- 6294385 TI - [The role of sympathetic beta-adrenergic receptor on renin secretion in essential hypertension]. PMID- 6294386 TI - [Evaluation of myocardial scintigraphy for the primary care of acute myocardial infarction]. PMID- 6294387 TI - [Classification of 201Tl-pertechnetate leg scan patterns and evaluation of their clinical significance]. PMID- 6294388 TI - [Effectiveness of single photon emission computed tomography in scanning of acute myocardial infarction with 99m Tc-PYP]. PMID- 6294389 TI - [Clinical evaluation of 67Ga and 201Tl scans in primary lung cancer]. PMID- 6294390 TI - Effect of cholecystokinin octapeptide and vasoactive intestinal polypeptide on adrenocortical secretion in the rat. AB - Intraperitoneal (i.p.) injection of C-terminal octapeptide of cholecystokinin (CCK-8) produced a dose-related increase in plasma corticosterone levels in intact rats, but not in vagotomized ones. Intracerebroventricular (i.c.v.) injection of CCK-8 was ineffective in stimulating the secretion of corticosterone, and in vitro experiment on ACTH release indicated that CCK-8 could not affect pituitary tissue directly. Since i.p. injection of non-sulfated CCK-8 failed to elevate plasma corticosterone levels, sulfated tyrosine residue in the CCK molecule is assumed to be indispensable for the stimulation of visceral organs. On the other hand, vasoactive intestinal polypeptide (VIP) was found to cause a dose-dependent increase in plasma corticosterone levels when administered centrally, but not after i.p. injection. However, VIP could not stimulate the release of ACTH from the pituitary tissue directly. The results suggest that VIP, but not CCK, stimulates the hypothalamic CRF neurons either directly or indirectly. PMID- 6294391 TI - Effects of beta-endorphin, thyrotropin-releasing hormone and cholecystokinin on body shaking behavior in rats. AB - Effects of intracerebroventricular administration of beta-endorphin, thyrotropin releasing hormone (TRH), cholecystokinin octapeptide (CCK-8), non-sulfated CCK-8 (CCK-8-NS) and caerulein on body shaking behavior were observed in rats. CCK-8 and its related peptides produced only a small increase in the number of body shakes, while TRH had the striking effect of stimulating body shakes, this increase being markedly suppressed by simultaneous administration of beta endorphin. Moreover, the suppressive effect of beta-endorphin on TRH-induced body shakes was antagonized by simultaneous administration of caerulein and CCK-8. The body shakes induced by ice-water immersion were also reduced by beta-endorphin, this beta-endorphin effect being partly antagonized by caerulein and CCK-8. PMID- 6294392 TI - Effects of 7-ethoxycarbonyl-6,8-dimethyl-4-hydroxymethyl-1(2H)-phthalazinone (EG626) on the spinal trigeminal nucleus, ventral posteromedial nucleus, and sensory cortex. AB - Effects of 7-ethoxycarbonyl-6,8-dimethyl-4-hydroxymethyl-1 (2H)-phthalazinone (EG626) on the spinal trigeminal nucleus (STN), ventral posteromedial nucleus (VPM), and sensory cortex were examined in cats anesthetized with alpha chloralose in comparison with the effects of morphine. EG626 produced a dose dependent inhibition of the polysynaptic components of the cortical field potentials upon VPM stimulation and either facilitatory or inhibitory effects on the polysynaptic components of the VPM field potential upon stimulation of the medial lemniscus, while the drug failed to affect the STN field potential with trigeminal nerve stimulation. Morphine inhibited the postsynaptic components of the STN field potentials and to a lesser extent, the polysynaptic components of the cortical field potential; and the effects of morphine on the VPM field potential were similar to those seen with EG626. Pretreatment of the animal with naloxone antagonized the facilitatory effect on the VPM field potentials produced by morphine, but not those by EG626. Morphine and EG626 induced either a prolonged increase in the blood flow or transient increase followed by a decrease in the blood flow in the VPM. These results suggest that EG626 may impair the polysynaptic transmission and/or neuron excitability in the sensory cortex and the VPM at least partly due to the change in blood flow there as does morphine. Unlike morphine, however, EG626 did not produce any obvious effect on the STN. PMID- 6294393 TI - Mechanism of relaxant action of papaverine. Effect on caffeine-induced contraction of guinea pig taenia coli. AB - The mode of action of papaverine on smooth muscle relaxation was examined through analyzing its effects on caffeine-induced contraction of guinea pig taenia coli, which might be due to calcium ion mobilized from intracellular store sites. Caffeine contraction induced in normal solution at 32 degrees C was decreased in the presence of papaverine but increased after the removal of papaverine, and it was also increased when the muscle was preincubated with papaverine and then washed out. These papaverine actions were inhibited at low temperature. Caffeine contraction induced in Ca-free, depolarizing solution was also enhanced by treatment with papaverine during Ca-loading. Cyclic AMP and its dibutyryl derivative produced a similar effect on the caffeine contraction induced in normal or Ca-free, depolarizing solution as those of papaverine. In addition, a good correlation was observed between the papaverine-induced increases in caffeine response and in tissue cyclic AMP level. From these results, it is considered that papaverine increases cyclic AMP of the taenia coli, and this increased cyclic AMP stimulates accumulation of calcium ion into its storage sites from where calcium ion was released by caffeine, finally leading to smooth muscle relaxation. PMID- 6294394 TI - Stimulation of renin release from dog renal cortical slices with L-isoproterenol and dibutyryl cyclic adenosine 3',5'-monophosphate. PMID- 6294395 TI - [Case of sclerosing hemangioma of the lung, pre-operative diagnosis by bronchial arteriography]. PMID- 6294396 TI - [Effect of dopamine on coronary circulation]. AB - The effect of dopamine on coronary and systemic circulation was examined in an experimental study on dogs using catheterization and extracorporeal perfusion of the bypassing branch of the left coronary artery, and simultaneous recording of heart's vascular resistance as well as cardio- and hemodynamic patterns. Dopamine introduced into coronary perfusion flow caused a dilatation of coronary vessels, a reflex drop in arterial blood pressure and peripheral vascular resistance, and changes in heart rate and intensity of cardiac contractions. Dopamine dosage should be about ten times as high to reproduce cardiodynamic and hemodynamic effects, comparable with the response to adrenaline and noradrenaline in terms of magnitude and pattern of development. Coronary vessels are dilated in two steps, through cholinergic activation and direct stimulation of beta-adrenoreactive systems. These two phases can be blocked by atropine and obsidan, respectively. After m-cholinergic and beta-adrenoreactive systems are blocked, dopamine produces a slight coronary vasoconstriction. PMID- 6294397 TI - [Dynamics of the cyclic nucleotide content in the acute period of myocardial infarct]. AB - Developing myocardial infarction is shown to be accompanied by raised plasma cAMP and cGMP levels which peak within the first few hours of the disease. Two patterns of changes were noted in the content of cyclic nucleotides: cAMP increase prevailing (a more typical pattern) and cGMP increase prevailing. Primary ventricular fibrillation was recorded in some patients belonging to the latter group. The development of cardiac failure is accompanied by a more stable rise of plasma cAMP. PMID- 6294398 TI - Diminished response of ovarian cAMP to luteinizing hormone in experimental uremia. AB - In prepuberal female rats with acute bilateral nephrectomy or chronic subtotal nephrectomy, the increase of ovarian cAMP concentration in response to submaximal doses of luteinizing hormone (LH 10 micrograms) and human chorionic gonadotropine (hCG 2.5 IU) was diminished (CO + 2.5 IU hCG 488 +/- 49 pmoles cAMP/mg protein; NX + 2.5 IU hCG 366 +/- 56. P less than 0.05). The cAMP response to follicle stimulating hormone (FSH) was unchanged. The abnormality was found both after administration of LH in vivo and incubation of ovaries with LH in vitro. Similarly, plasma estradiol concentrations in response to submaximal hCG stimulation were diminished. Basal cAMP concentrations and cAMP concentrations after maximal stimulation were unchanged. The defect was observed both in ovaries of untreated prepuberal rats, of pregnant mare serum (PMS)-treated rats (follicular phase) and PMS/hCG-treated rats (luteal phase). Diminished ovarian cAMP response to LH was observed both in parathyroid intact and in parathyroidectomized rats. Administration of 1,25(OH)2D3 in physiological doses (60 ng/kg) to acutely uremic rats restored diminished ovarian cAMP response to submaximal LH stimulation irrespective of parathyroid status. The effect of 1,25(OH)2D3 could not be reproduced by hypercalcemia resulting from intraperitoneal calcium injection. In vivo administration of indomethacin further diminished ovarian cAMP response in uremic animals and had no effect in control animals. Incubation of ovaries with PGE1 and PGE2 increased basal and stimulated cAMP concentrations and abolished the difference between control and uremic animals. The diminished response of ovarian cAMP content to submaximal doses of hCG was not corrected by bromocriptine (1 mg/kg) despite normalization of hyperprolactinemia. The present study shows diminished ovarian cAMP and plasma estradiol response to LH in experimental uremia. It documents a role of 1,25(OH)2D3 and prostaglandins in the genesis of this abnormality. PMID- 6294399 TI - Action of phosphorus on calcium release in isolated perfused rat tails. AB - A bone perfusion system using isolated rat tails was developed to study the action of inorganic phosphorus (Pi) on calcium (Ca) release in bone. Rats were either pre-labelled with 45Ca to study calcium removal from bone or 45Ca was added to the perfusate to study Ca deposition into bone. When perfusate Pi was increased from 3.5 to 5.25 mg/dl, a marked inhibition of 45Ca release was demonstrated. 85Sr microspheres, used to demonstrate differences in the flow of perfusate between bone and soft tissue, showed that the Pi effect was not due to alterations in perfusate flow to bone. Increasing the perfusate Pi concentration from 3.5 to 7 mg/dl did not increase 45Ca deposition into bone, nor did it affect baseline or PTH-stimulated cyclic AMP production. The Pi effect was inhibited by reducing the perfusate temperature to 4 degrees C, by using an anerobic perfusate, adding iodoacetate to the perfusate, and finally, by pre-treating the rats with actinomycin D prior to the experiment. Thus, in this model an increase in the Pi concentration of the perfusate inhibits Ca release from bone but does not increase Ca uptake by bone. The inhibitory effect appears to be dependent on normal cell metabolism. PMID- 6294400 TI - [Role of endolymphatic antibacterial therapy in the preparation of patients for pulmonary surgery]. PMID- 6294401 TI - [Benign lung tumors]. PMID- 6294402 TI - [Radionuclide diagnosis of internal hemorrhage in abdominal diseases and injuries]. PMID- 6294403 TI - [Glue hemostasis in gastrointestinal surgery]. PMID- 6294404 TI - Use of enzyme-linked immunosorbent assay for detection of IgM antibodies to Japanese encephalitis virus in swine sera. PMID- 6294405 TI - LDL receptor activity of cultured arterial smooth muscle cells from genetically hyperlipidemic rabbits (WHHL-rabbit). PMID- 6294406 TI - [Clinical forms of herpetic meningoencephalitis in adults]. PMID- 6294408 TI - Genetic analysis of polykaryocytosis by herpes simplex virus. V. An ultraviolet light-induced r mutant which probably possesses a suppressor gene. PMID- 6294407 TI - [Energy reactions in the skeletal muscles of rats following space flight on the Kosmos-936 biosatellite]. AB - The respiration of mitochondria isolated from mixed skeletal muscles of hindlimbs of rats flown for 18.5 days on Cosmos-936 was investigated polarographically. At R + 10 hours the rate of mitochondrial respiration in different metabolic states during the oxidation of succinic acid and NAD-dependent substrates declined. The enzyme activity of mitochondrial cytochrome oxidase and cytosol lactate dehydrogenase diminished. At R + 25 days both aerobic and anaerobic oxidative processes increased, thus leading to the recovery of the parameters (sometimes they not only returned to the norm but exceeded it). PMID- 6294410 TI - Identification of basic drugs in urine by dual fused silica capillary column GC. AB - A method for analyzing urine for the presence of basic drugs and their metabolites using two fused silica capillary columns is described along with operating conditions and column characteristics. Urine extracts were injected into a gas chromatograph equipped with nitrogen phosphorous detectors (GC/NPD) and two capillary columns, housed in a single injection port. The injected material was simultaneously chromatographed in each column. Identification of unknowns was done using relative retention times. PMID- 6294409 TI - Use of cerebellar cGMP in the measurement and characterization of the diazepam/ethanol interaction. AB - The characterization and measurement of the benzodiazepine and ethanol interaction has been of major interest for many years. Various pharmacological and biochemical studies have been employed to investigate this interaction, which is believed to occur predominantly in the CNS, but localization of one particular brain area has not been investigated. This research employed the rat cerebellum as a site to study the diazepam/ethanol interaction. The measurement of cerebellar ethanol and diazepam, by gas chromatography, demonstrated an enhancement of diazepam levels by ethanol. Ethanol and diazepam, alone and in combination, displayed a significant depression of cerebellar 3',5'-Guanosine Cyclic Monophosphate (cGMP). The depression seen by the combination was significantly greater than the simple algebraic sum, but insignificantly different from the corrected algebraic sum. The double reciprocal plot of the data demonstrated a common ordinate intercept for the diazepam line and the diazepam/ethanol (2 g/kg) line, thus indicating a competitive mechanism of action. PMID- 6294411 TI - Androgen receptors in fetal rabbit lung and the effect of fetal sex on the levels of circulating hormones and pulmonary hormone receptors. AB - High affinity (KD = 0.2 nM), low capacity (3.6-5.0 fmol/mg protein), androgen specific binding proteins with characteristics typical of androgen receptors were identified in the lungs of rabbit fetuses between the 26 and 29th day of gestation and in the lungs of adult rabbits. While androgen receptor concentrations increased significantly from late gestation to adulthood (P less than 0.01), no sex-related differences were observed in either the binding affinities or concentrations of the receptors at any age tested. Similarly, no sex-related differences were found in the levels of progesterone, cortisol and cortisone in the fetal circulation, or in the levels of progesterone receptors, glucocorticoid receptors and beta-adrenergic receptors in the fetal lung at 26 days of gestation. It is concluded that the fetal lung interacts directly with circulating androgens via specific androgen receptors and that the suggested male disadvantage with respect to lung maturation in the perinatal period does not appear to be associated with sex-related differences in the levels of pulmonary androgen, glucocorticoid, progesterone or beta-adrenergic receptors. PMID- 6294412 TI - A proton magnetic resonance study of the 18-hydroxy derivatives of cortisol and corticosterone. AB - Proton magnetic resonance spectroscopy played a crucial role in the identification of a new corticosteroid, 18-hydroxycortisol, recently isolated from the urine of a patient with an aldosterone-producing adenoma. Mass spectrometric analysis and chemical degradative studies demonstrated an empirical formula of C21 H30 O6 corresponding to a diketopregnenetetrol and placed three of the four hydroxyl groups at the 11 beta, 18 and 21 positions. The first suggestion that the locus of the fourth hydroxyl was 17 alpha came from the n.m.r. spectrum in the form of negative evidence for proton linked to the carbon atom bearing this fourth hydroxyl. The n.m.r. spectral features of the 18-hydroxy derivatives of cortisol and corticosterone are compared. Both were found to exist in the 20, 18 cyclic hemiketal form. PMID- 6294413 TI - Characteristics of the response of dispersed guinea-pig adrenal cells to low doses (1-50 pg/ml) of alpha 1-24 adrenocorticotrophin. AB - Isolated adrenal cells prepared by tryptic digestion of the guinea-pig adrenal gland are sensitive to low concentrations (less than 25 pg/ml) of adrenocorticotrophin (ACTH). Cell which have been pre-incubated for 2 h. centrifuged and resuspended in fresh culture medium prior to the introduction of 10 pg/ml ACTH for 60 min show a marked increase (328 +/- 109 nmol/l; mean +/- SD) in cortisol secretion over the control compared to freshly dispersed cells (75 +/ 45 nmol/l). Further potentiation of the ACTH effect was seen with the pre incubated cells by suplementing the medium with calcium (8 mM) and ascorbate (2 mM) but not with theophylline (1 mM). Basal cortisol secretion was not affected by any of the additives. In the presence of 8 mM calcium and after 60 min incubation 10 pg/ml ACTH stimulated cortisol secretion from 328 nmol/l over the control to 839 +/- 382 nmol/l. The effect of ascorbate (2 mM) was to further increase the effect of ACTH at all dose levels tested (1-25 pg/ml). The concentration of ACTH required to provoke half maximal cortisol secretion decreased from 95 pg/ml with normal medium to 12 pg/ml with calcium -ascorbate supplemented medium. Using this supplemented medium the cells were sensitive to 1 pg/ml and cortisol secretion was stimulated 10-fold over the control with 50 pg/ml, a dose which saturated the system. PMID- 6294414 TI - Synthesis and biological activity of 3 beta-hydroxy-9,10-secopregna-5,7,10[19] triene-20-one: a side chain analogue of vitamin D3. AB - The synthesis, biological and antagonistic activity of 3 beta-hydroxy-9,10 secopregna-5,7,10[19]-triene-20-one (20-oxopregnacalciferol, 7) a shortened side chain analogue of vitamin D3, are described. At the highest dose tested the analogue was found to have small though significant bone and soft tissue mobilization activity; no significant increase in intestinal calcium transport was noted. The compound was found to possess no antagonistic activity against vitamin D3. PMID- 6294415 TI - Changes in corticosteroid secretory pattern induced by prolonged corticotropin treatment in the rabbit. PMID- 6294416 TI - The average spacing of restriction enzyme recognition sites in DNA. PMID- 6294417 TI - Capping and mitogenesis: a model implicating microfilaments in lymphocyte activation. PMID- 6294418 TI - Adrenocortical hormone levels during cardiopulmonary bypass with and without pulsatile flow. AB - To determine the effect of hypothermic pulsatile and nonpulsatile cardiopulmonary bypass (CPB) with hemodilution on adrenocortical function we measured plasma levels of adrenocorticotropic hormone (ACTH), cortisol, aldosterone, and renin in two groups of patients. Group I, comprising 11 patients had routine CPB (nonpulsatile), and Group II, comprising 12 patients, had pulsatile flow during CPB (pulsatile). Both groups demonstrated comparable increases in cortisol, ACTH, and aldosterone with operation. Levels for all three hormones appeared to decline during CPB and then rose again in the post-CPB period. There were no significant differences between groups. Plasma renin activity gradually declined in a comparable manner in both groups. In the post-CPB period, renin activity was slightly higher in the nonpulsatile group (1.7 +/- 0.5 versus 0.8 +/- 0.2 ng/ml/hr, p less than 0.05). Correction for the effect of hemodilution demonstrated no decrease in cortisol and a slight increase in ACTH in both groups during CPB. Significant increases occurred in both groups during CPB in urinary Na+ excretion rate and urinary Na+/K+ ratio, more so for the nonpulsatile group. There was no correlation between urinary Na+/K+ ratios and either plasma cortisol or aldosterone levels. Thus routine CPB demonstrates no evidence of adrenocortical hypofunction and the addition of pulsatile flow produces little improvement. PMID- 6294419 TI - Encephalomyocarditis virus as a probe of errors in macromolecular synthesis in aging mice. AB - A viral probe was used in attempts to develop an in vivo test of the hypothesis that cellular senescence is accompanied by an increased rate of errors in macromolecular synthesis. Young and senescent Balb/cNNia mice were infected with encephalomyocarditis (EMC) virus. No differences in pattern of infection or titers of virus in brain and heart were observed between the two age groups. The yield of virus in control experiments was shown to be reduced by growth in the presence of 5-fluorouracil or a mixture of three amino acid analogs. Since the growth of this virus is highly dependent upon host cell synthetic machinery, these results are thought to suggest that substantial elevations in the rate of errors in macromolecular synthesis in these tissues do not occur with age. Further studies might allow a more precise determination of whether there is an age correlation of in vivo error rates; for the EMC virus, a selectable marker suitable for the quantitation of rates of mutation in vivo has so far not been obtainable. PMID- 6294420 TI - [Treatment of anaplastic carcinoma of the lung with VP 16-213]. PMID- 6294421 TI - [Acute respiratory failure as the initial manifestation of hepatocellular carcinoma. Apropos of 2 cases]. PMID- 6294422 TI - Mechanism of photoaffinity labeling of P2-purinergic receptors by arylazido aminopropionyl atp in isolated guinea-pig vas deferens. PMID- 6294423 TI - Regional heterogeneity of benzodiazepine receptors at 37 degrees C: an in vitro study in various regions of the rat brain. AB - The most compelling pharmacological evidence in support of benzo-diazepine (BZD) receptor heterogeneity is derived from the study of the complex interactions of CL 218872 and propyl beta-carboline-3-carboxylate (PCC) with brain BZD receptors. In the present study, we provide evidence to support the hypothesis that intraregional BZD receptor heterogeneity in rat brain is a result of the different conformational states of a single receptor. This hypothesis is based upon the observation that CL 218872 and PCC lose the ability to effectively discriminate BZD receptor subtypes in rat cerebral cortex, hippocampus and pons medulla at physiological temperature (37 degrees C). Interestingly, both PCC and CL 218872 show higher affinity for BZD receptors in the cerebellum when compared to other brain regions at 37 degrees C. This observation suggests that interregional BZD receptor heterogeneity occurs under physiologically relevant temperatures. We propose that distinct cerebellar and non-cerebellar type BZD receptors exist in vivo while marked differences in the affinity of the type I and type II BZD receptor subtypes postulated by Klepner et al. 1979 may only occur in vitro at 0 degree--4 degree C. PMID- 6294424 TI - A comparison of the receptor constants of morphine and ethylketocyclazocine for analgesia and inhibition of gastrointestinal transit in the rat. AB - The efficacies and dissociation constants of proposed mu and kappa receptor agonists (morphine and ethylketocyclazocine, respectively) were compared using the method of partial irreversible blockade (with buprenorphine) and Stephenson's theory of drug action. While there was good agreement between the dissociation constant (KA) of morphine in analgesia (3.3 x 10(-5) M) and in inhibition of gastrointestinal transit (1.1 x 10(-5) M), the KA of ethylketocyclazocine differed by an order of magnitude in these endpoints (3.2 x 10(-6) M and 6.7 x 10(-5) M, respectively). The efficacies of morphine were found to be similar for the two effects studied (4.23 and 5.26), while those for ethylketocyclazocine differed markedly (2.06 and 10.39). The fraction of receptors remaining unblocked after buprenorphine was consistent for the test but not for the agonist, indicating a different distribution of receptors for the two endpoints. Our results strongly suggest that morphine induces analgesia, and slows transit in the small intestine, through the same type of receptor. The same conclusion cannot be drawn for ethylketocyclazocine. PMID- 6294425 TI - Schizophrenia and reduced cyclic AMP production: evidence for the role of receptor-linked events. AB - Reduced cyclic AMP (cAMP) production has been found in platelets of schizophrenic patients. cAMP is generated physiologically as a result of a series of steps beginning with receptor activation by a ligand, progressing through activation of the enzyme protein, adenylate cyclase. The deficit of cAMP found in the schizophrenic population may occur at any one, or at multiple steps in this cascade. The present study attempts to discriminate whether impaired adenylate cyclase itself was responsible for the cAMP deficit or whether abnormalities in receptor events or linkage are present in schizophrenics. The production of cAMP following direct stimulation of adenylate cyclase by NaF was contrasted with receptor mediated activation of adenylate cyclase by prostaglandin E1 (PGE1) in disrupted platelet preparations from schizophrenics and normal controls. cAMP formation stimulated by NaF was not different in platelets of schizophrenics as compared to controls, however, platelets of schizophrenics showed reduced response to PGE1 stimulation. The authors interpret these findings as evidence for a membrane associated abnormality of either receptor or receptor-adenylate cyclase linkage in the schizophrenias. PMID- 6294426 TI - HL 725, an extremely potent inhibitor of platelet phosphodiesterase and induced platelet aggregation in vitro. AB - The new pyrimido-isoquinoline compound HL 725 is an extremely potent inhibitor of the aggregation of human platelets induced in vitro by ADP, collagen, thrombin and epinephrine. The aggregation induced by 0,5 mM arachidonic acid is inhibited about 50% with 50 pM HL 725. Thus the potency of HL 725 is higher than that of prostacyclin, the most active natural inhibitor of aggregation. We hypothesize that HL 725 inhibits the enzymatic degradation of cyclic adenosine 3', 5' monophosphate (cAMP) in the platelets. In accordance with this proposal is the strong inhibitory action on cAMP phosphodiesterase extracted from human platelets. About 250 pM HL 725 inhibited 50% of the activity of this enzyme at a substrate concentration of 0.5 microM. A marked elevation of cAMP levels in human platelets could be demonstrated after incubation in vitro with 100 nM HL 725. PMID- 6294427 TI - L-dopa mediated accumulation of cyclic AMP in isolated rabbit retinae in vitro. Effects of light and/or pharmacological factors. AB - The ability of L-dopa to stimulate the formation of cyclic AMP in pieces of intact rabbit retina in vitro has been studied and compared with that of dopamine and of other dopamine-mimetic drugs. Dose-dependent effects were measured in response to 5 microM up to 100 microM L-dopa with a maximal stimulation after 20 min of incubation at 35 degrees C. The L-dopa precursor, tyrosine, was totally ineffective. The L-dopa mediated response was detectable only in pieces of intact tissues (and not in homogenates) and is presumably due to the formation of newly formed dopamine, since it was completely inhibited by a decarboxylase inhibitor (benserazide). The biochemical response (cyclic AMP increase) was facilitated by ambient light, this effect being potentiated by 56 mM K+ or fully inhibited by 1 mM gamma-butyrolactone (GBL). The data suggest that the measurement of cyclic AMP levels in pieces of rabbit retina may provide a useful neurochemical model for the study of physiological and/or pharmacological agents able to interact at pre- and/or post-synaptic dopaminergic sites. PMID- 6294428 TI - Increase of beta-endorphin concentrations in the plasma and pituitary neuro intermediate lobe of the rat on the afternoon of proestrus. AB - Beta-endorphin (beta-EP) concentrations in the plasma and the anterior and neuro intermediate lobes of the pituitary (AP and NIL) were quantitated by radioimmunoassay (RIA) and gel filtration chromatography in female rats at 1000, 1400, and 1900 h on the day of proestrus and diestrus day-1. There were no significant changes in beta-EP in the plasma, AP, or NIL on diestrus day-1. On proestrus, beta-EP in the plasma and NIL, but not the AP, increased significantly from 1000 to 1400 h and returned to basal levels by 1900 h. The time course of this increase of beta-EP in the NIL and plasma is consistent with the temporal sequence of the prolactin and gonadotropin surges on the afternoon of proestrus, suggesting that beta-EP in the NIL may be involved in the regulation of these neuroendocrine events. PMID- 6294429 TI - CRF stimulates alpha-MSH secretion and cyclic AMP accumulation in rat pars intermedia cells. PMID- 6294430 TI - Anti-lewisite activity and stability of meso-dimercaptosuccinic acid and 2,3 dimercapto-1-propanesulfonic acid. AB - Meso-dimercaptosuccinic acid (DMSA) and the sodium salt of 2,3-dimercapto-1 propanesulfonic acid (DMPS) are analogous in chemical structure to dimercaprol (BAL, British Anti-Lewisite). Dimercaprol was among the first therapeutically useful metal chelating agents and was developed originally as an anti-lewisite agent. Either DMSA or DMPS protects rabbits from the lethal systemic action of dichloro(2-chlorovinyl)arsine (29.7 mumols/kg, also known as lewisite. The analogs are active in this respect when given either sc or po. The stability of each of the three dimercapto compounds in distilled H2O, pH 7.0 at 24 degrees, has been examined for seven days. DMSA retained 82% of its mercapto groups, but no titratable mercapto groups remained in the DMPS or BAL solutions. At pH 5.0, however, there was no striking difference in the stability of the three dimercapto compounds (78-87%) over a seven day period. DMSA and DMPS warrant further investigation as water soluble metal binding agents in both in vivo and in vitro experiments. PMID- 6294431 TI - GABAergic control of anterior pituitary hormone secretion. AB - Anatomical and biochemical studies have identified a hypothalamic tubero infundibular GABAergic system, which plays a functional role on anterior pituitary hormone secretion. Experimental and clinical evidence support the presence of a dual component in the action of GABA; one mediated via the central nervous system and the other exerted directly at the anterior pituitary level. The two sites of action may be responsible for the excitatory and inhibitory effects of GABA on pituitary hormone and especially prolactin secretion. The future characterization of this system will provide a better understanding of the involvement of GABA in the physiology of anterior pituitary hormone secretion and will contribute to the development of new pharmacological agents for the therapy of neuroendocrine disorders. PMID- 6294432 TI - Tolerance and cross tolerance with morphine resulting from physiological release of endogenous opiates. AB - Mice which had been exposed to a chronic schedule of warm water swimming showed the development of a significant tolerance to the antinociceptive response (tail flick latency) and a significant, two-fold increase in the ED50 of morphine (tail flick latency and abdominal constriction response). These results suggest the involvement of endogenous opiates during swim stress in mice and are consistent with the hypothesis that during chronic stress the opiate receptors are activated in a manner analogous to the repeated application of exogenous opiates producing tolerance, morphine cross tolerance and (as previously reported) withdrawal-like behaviour. PMID- 6294433 TI - Opiate receptor blockade in man reduces 2-deoxy-D-glucose-induced food intake but not hunger, thirst, and hypothermia. AB - Opioid peptides may act as neuromodulators in the central nervous system to conserve energy stores and water in mammals. To examine this hypothesis in man, the effect of opiate receptor blockade with naloxone on the hunger, thirst, and hypothermic response to 2-deoxy-D-glucose-induced glucoprivic stress was assessed. Opiate receptor blockade decreased stress-induced food intake but did not reduce marked increases in hunger produced by glucoprivation. Naloxone infusions did not change the hypercortisolemic, polydipsic, hypothermic, and thermogenic response to 2-deoxy-D-glucose. While these results do not suggest a major role for a beta-endorphin modulation of stress-induced hunger, hypothermia and water conservation, the reduction of food intake could be due to augmented satiety, perhaps associated with retardation of gastric emptying during opiate receptor blockade. PMID- 6294434 TI - Preferential oxidation of linolenic acid compared to linoleic acid in the liver of catfish (Heteropneustes fossilis and Clarias batrachus). AB - The fate of [1(-14)C] linoleic acid and [1(-14)C] linolenic acid in the liver slices and also in the liver tissues of live carnivorous catfish, Heteropneustes fossilis and Clarias batrachus, was studied. Incorporation of the fatty acids into different lipid classes in the live fish differed greatly from the tissue slices, indicating certain physiological control operative in vivo. The extent of desaturation and chain elongation of linoleic and linolenic acids into long-chain polyunsaturated fatty acids was low. Linolenic acid was oxidized (thus labeling the saturated fatty acid with liberated 14C-acetyl-CoA) in preference to linoleic acid, and this oxidation also seemed to be under physiological control since both of the fatty acids were poorly oxidized in the tissue slices and in the killed fish. These fish can therefore recognize the difference in the acyl chain structures of linoleate and linolenate. The higher oxidation of linolenic acid and poor capacity for its conversion to longer chain, highly unsaturated derivatives indicates a higher demand for the dietary supply of these essential fatty acids in these two species. PMID- 6294435 TI - Sclerosing hepatic carcinoma: relationship to hypercalcemia. AB - A series of 30 patients is reported whose primary hepatic tumors had a distinctive histologic pattern that we have called "sclerosing hepatic carcinoma" (SHC). Sixty-nine percent of those tested had hypercalcemia and low levels of serum phosphate. As comparison, 38 control patients who had either classical peripheral cholangiocarcinoma or typical hepatocellular carcinoma were studied. In the control group, only two patients, who also had bone metastases, had hypercalcemia. Sclerosing hepatic carcinoma is characterized by intense fibrosis in which the tubular neoplastic structures are embedded. Although the tumor in each patient superficially resembled peripheral cholangiocarcinoma, on close inspection 63% were found to be of apparent hepatocyte origin, 20% were apparently ductal, and 13% were mixed or not distinguishable. One patient's tumor had the pattern of the rare cholangiolocellular carcinoma. The difficulty of histological diagnosis was well illustrated by the fact that none of premortem biopsies in 16 patients was correctly interpreted unequivocally as primary carcinoma of the liver. Many were misinterpreted as metastatic adenocarcinomas, most frequently of pancreatic origin. We believe that, by describing clinicopathological features, more attention will be drawn to this unique carcinoma of liver origin. PMID- 6294436 TI - Ground glass inclusions in liver cells in an alcoholic treated with cyanamide (Dipsan). PMID- 6294437 TI - Albumin receptors on hepatitis B virus and antibodies to albumin. PMID- 6294438 TI - The hepatitis A virus infection. PMID- 6294439 TI - Non-inflammatory herpes simplex hepatitis in an adult with chronic neutropenia. AB - We report the case of a young woman with chronic neutropenia, in whom hepatitis, extensive herpetic eruption and herpes simplex viremia developed after genital herpetic ulceration. Although severe liver necrosis was present, the patient's death did not result from hepatic failure. No inflammatory cell infiltration was found circumscribing the multiple necrotic foci in the liver. This absence of inflammatory cell infiltration reflects the host's inability to normally restrain herpes simplex virus dissemination and, in this patient, might be the consequence of chronic neutropenia. PMID- 6294440 TI - Invasion of the lumen of the bile ducts by hepatocellular carcinoma. AB - The purpose of this investigation was to ascertain the prevalence and manifestations of tumorous invasion of the lumen of the bile ducts, a mode of local extension characteristic of hepatocellular carcinoma. In a series of 140 necropsied patients with hepatocellular carcinoma, tumorous invasion of the bile duct was noted in three patients (2.1%). Marked cholestasis was present in these three patients; the other manifestations related to the tumorous invasion of the bile duct were biliary pain in one and gall-bladder enlargement in two patients. PMID- 6294441 TI - A clinical and pathological study of diffuse type hepatocellular carcinoma. AB - Six autopsy cases of the rare, diffuse type of hepatocellular carcinoma (HCC), as classified gross anatomically according to the strict definition, have been studied. The prominent clinical feature was the rapid deterioration of the patient's general condition, terminating in hepatic failure. The liver size enlarged quickly, at a perceptible speed, often accompanied by abdominal pain. Diagnosis of this particular type of HCC was difficult, and celiac angiography and scintiscan of the liver were only suggestive when considered together with other laboratory data. Hepatitis B surface antigen was positive in all three patients in whom it was tested. The entire liver was studded with minute, uniformly sized tumor nodules, evenly distributed throughout. Some of them were grossly indistinguishable from cirrhotic nodules. All livers had an underlying cirrhosis which was characterized by relatively small regenerative nodules with thin stromas. Large portal branches at the hilum contained tumor thrombi in all patients, except for one case in which left lobectomy was followed by intraportal dissemination. Histologically, all tumor nodules represented intrahepatic metastases via the portal vein system. Tumor cells were poorly differentiated. These findings suggest that the diffuse type of HCC most frequently, if not always, represents intrahepatic, widespread portal metastases which have occurred within a short period of time. PMID- 6294442 TI - Hepatitis B virus antigens in liver tissue in hepatocellular carcinoma and advanced chronic liver disease-relationship to liver cell dysplasia. AB - Hepatitis B surface (HBs) and core (HBc) antigens (Ag) were studied in liver tissue in HBsAg seropositive patients with chronic liver disease complicated (n = 32) and not complicated (n = 36) by hepatocellular carcinoma. Both groups were matched by age, sex and underlying disease. There was no qualitative and quantitative difference in tissue HBsAg between the two groups. However, HBcAg was significantly less in quantity in hepatocytes of patients with hepatocellular carcinoma compared to chronic liver disease without cancer. Serum hepatitis B e antigen tested by radioimmunoassay was also less frequently positive in patients with hepatocellular carcinoma. These findings seem to suggest that hepatitis B virus replication becomes less active in the process of hepatocarcinogenesis. The relationship between intrahepatic hepatitis B antigens and liver cell dysplasia was also studied. In hepatocellular carcinoma, tissue hepatitis B antigens often coexisted in the same liver having liver cell dysplasia, but no such association was observed in chronic liver disease without cancer. However, no indication was obtained that the dysplastic cells harbor HBsAg more frequently than non dysplastic cells. PMID- 6294443 TI - Immunocytochemical study of pyruvate kinase isoenzymes in normal and pathologic human liver. AB - In healthy human livers, L pyruvate kinase (L PK) was detected by immunofluorescence and double labelling in hepatocytes and M PK was detected in bile duct epithelial cells. Numerous associations between isoenzyme type and hepatological lesion were observed. In 10 cases of ethanolic cirrhosis, L and M PK were simultaneously observed in the hepatocytes of regenerative nodules, as they were in biliary neoductules; fibrotic regions were L and M PK negative and the hepatocytes in the anastomosing plate system were almost exclusively L PK positive, as in normal subjects. In 12 hepatocarcinomas, cancer cells had a double L and M specificity with variations in the intensity of the M isoenzyme, while the stroma reaction was L and M negative. In five hepatoblastomas, the simultaneous presence of L and M PK was also observed, but the consistently marked intensity of M PK argues in favour of the embryonic nature of this type of cancer. The results suggest that mature, highly differentiated cells exclusively and specifically synthesize one isoenzyme. A double specificity in pathology reflects the dysfunctional state of the hepatic and biliary epithelial cells or a dedifferentiated state which may terminate with the appearance of a cancer. PMID- 6294444 TI - [Addison crisis in acute kidney failure]. PMID- 6294445 TI - [Infections causes of congenital abnormalities]. PMID- 6294446 TI - Status of the red cell Na,K-pump in hyper- and hypothyroidism. AB - To investigate the status of the sodium-potassium pump in cells of human subjects with abnormal thyroid function, we measured the number of pump units as well as the cation transport activity of the pump in erythrocytes from 23 hyperthyroid and 7 hypothyroid patients. It was found that the number of Na+-K+-ATPase units in erythrocytes (as measured by ouabain binding) was significantly reduced in hyperthyroidism (mean 36% below controls, p less than 0.001). The rate of rubidium uptake by the same cells was also reduced, but to a smaller extent (mean 9%, p less than 0.02). These changes were reversible with control of the hyperthyroidism. Hypothyroid individuals showed changes in erythrocyte Na+-K+ pump which were in the opposite direction to those seen in hyperthyroidism. It is concluded that thyroid hormone exerts a marked negative influence on the number of Na+-K+ pump units in one easily available human cell type. The direction of the effect suggests a complex relationship between thyroid hormone and the level of the Na,K-ATPase in any one tissue. Whatever the cellular mechanism responsible for the effects observed in the red cell, these changes should provide a measure of thyroid hormone action at a cellular level and this may prove useful in the study of thyroid hormone physiology in man. PMID- 6294447 TI - NMR and EPR studies of chromium and cobalt nucleotides and their interactions with enzymes. PMID- 6294449 TI - Galactose-1-phosphate uridylyltransferase: detection, isolation, and characterization of the uridylyl enzyme. PMID- 6294448 TI - Stereochemistry of enzymic phosphoryl and nucleotidyl transfer. PMID- 6294450 TI - Stereochemistry of selected phosphotransferases and nucleotidyltransferases. PMID- 6294451 TI - Chiral [16O, 17O, 18O]phosphoric monoesters as stereochemical probes of phosphotransferases. PMID- 6294452 TI - The stereochemical course of nucleoside triphosphatase reactions. PMID- 6294453 TI - Kinetic examination of enzyme mechanisms involving branched reaction pathways--a detailed consideration of multifunctional glucose-6-phosphatase. PMID- 6294454 TI - Intermediates in the phosphoglycerate mutase and bisphosphoglycerate synthase reactions. PMID- 6294455 TI - Two-protonic-state electrophiles as probes of enzyme mechanisms. PMID- 6294456 TI - Covalent chemistry of pyruvate, orthophosphate dikinase. PMID- 6294457 TI - Solvent isotope effects of enzyme systems. PMID- 6294459 TI - Enzymology of enol intermediates. PMID- 6294458 TI - Carbonic anhydrase: oxygen-18 exchange catalyzed by an enzyme with rate contributing proton-transfer steps. PMID- 6294460 TI - Intermolecular and intramolecular transposition and transposition immunity in Tn3 and Tn2660. AB - Intermolecular transposition of Tn2660 into pCR1 was measured at 30 degrees C in recA- and recA+ hosts as between 2.6 and 5.5 X 10(-3), a similar value to that previously found for Tn3. No cointegrate structures were found under conditions where 10(4) transposition events occurred. Immunity to intermolecular transposition of Tn2660, similar to that found for Tn3 was demonstrated by showing that the above transposition frequency was reduced by a factor of between 10(-3) and 10(-4) when a mutant Tn2660 (resulting in the synthesis of a temperature-sensitive beta-lactamase) was present in the recipient plasmid. Intramolecular transposition of Tn3 was found to occur under the same conditions as previously demonstrated for Tn2660 giving rise to similar end products, in which the newly introduced Tn3 is oriented inversely to the resident Tn3 and the DNA sequence between the two transposons has been inverted. Thus, in all respects functional identity of the transposition activities of Tn3 and Tn2660 is shown, thereby identifying characteristics of intramolecular transposition that are not readily accommodated by current models of transposition. PMID- 6294461 TI - Genetic evidence that control of F replication is negative. AB - We have taken advantage of two situations in which the incompatibility barrier between F plasmids is overcome to show that wild-type genes controlling F copy number (cop+) are dominant in trans over mutant genes. The simplest interpretation of our findings is that the cop mutations have inactivated a repressor gene that controls F replication. Since the cop. mutations all map in a region that others have shown by sequence analysis to theoretically encode four proteins, a strong possibility exists that one of these proteins is the repressor. PMID- 6294462 TI - Isolation of a recombinant lambda phage carrying nusA and surrounding region of the Escherichia coli K-12 chromosome. AB - A recombinant bacteriophage lambda, lambda argG-6, has been isolated which carries the argG gene and neighbouring loci on an EcoRI-generated 15.5 Kb DNA fragment from the Escherichia coli chromosome. The locations of the argG, nusA and pnp genes on the 15.5 Kb DNA fragment have been determined. In the case of nusA, a Tn5 insertion and sub-cloning of restriction fragments were used to locate the gene. The gene products of nusA and pnp have been identified on one- and two-dimensional polyacrylamide gels. The clockwise gene order was found to be argG-nusA-pnp. PMID- 6294463 TI - Gene expression in Streptomyces: construction and application of promoter-probe plasmid vectors in Streptomyces lividans. AB - Promoter-probe plasmid vectors were constructed for Streptomyces lividans using expression of the Escherichia coli chloramphenicol acetyltransferase gene as an indicator of promoter activity. These vectors have been used to isolate and to study the activity of DNA sequences that contain transcriptional control signals from Streptomyces, Bacillus licheniformis, E. coli, and Serratia marcescens. Studies of these promoter regions in heterospecific hosts indicate that genus or species-specific factors may present barriers to the expression of bacterial genetic material in certain heterologous cellular environments. While promoter regions isolated from E. coli, S. marcescens and B. licheniformis all appear to be recognized by the RNA polymerase of S. lividans, the Streptomyces transcriptional control signals isolated do not appear to function normally in E. coli. PMID- 6294464 TI - crpX mutants of Escherichia coli K12: specific regulatory effects of altered cyclic AMP receptor proteins. AB - We attempted to correlate structural modifications of the adenosine 3',5' cyclic monophosphate (cAMP) receptor protein (CAP), to changes in some of its in vivo regulatory functions such as (i) stimulation of the lactose operon expression and (ii) control of adenylate cyclase activity. A radioimmunological procedure was used to study the structure of CAP synthesized by three mutants (crpX) grown under various conditions, in the presence or absence of endogenous or exogenous cAMP. In one mutant CAP appears to be sensitive to thermal inactivation. In another mutant CAP is particularly sensitive to degradation in the absence of cAMP; this degradation is enhanced by high temperature and during stationary phase of growth, and prevented by the addition of glucose. Functional alterations of CAP were not found to follow structural changes strictly. In the crpX mutants and in strains carrying the crp+ or other crp allele, the stimulation of the lactose operon expression and the modulation of the in vivo rates of cAMP synthesis appear to vary in parallel, favoring an indirect mechanism of regulation of adenylate cyclase by CAP. PMID- 6294465 TI - Sequence relationships between plasmids associated with conventional MLS resistance and zonal lincomycin resistance in Streptococcus pyogenes. AB - By using electron microscopy of self-annealed DNA and restriction enzyme analysis, we have compared the physical maps of two group A streptococcal plasmids associated with conventional MLS resistance (pEL1; 20 Md) and zonal lincomycin resistance (pSM10419; 15 Md). Of their monomeric molecules, about 40% and 60%, respectively, are occupied by identical non-tandem inverted repeats containing sequences specifying putative replication functions. Sequence homology also exists between their resistance determinants which are located in unique DNA. Moreover, homology between additional regions of unknown function is so extensive and restriction fragment arrangement so similar that, formally, pSM10419 can be considered a deletion variant of pEL1. The results suggest that MLS and zonal lincomycin resistance have the same biochemical basis (i.e. methylation of 23S ribosomal RNA) and differ only quantitatively in the inducible control systems. PMID- 6294466 TI - Construction of a Schizosaccharomyces pombe gene bank in a yeast bacterial shuttle vector and its use to isolate genes by complementation. AB - A gene bank of partial Sau3A restriction fragments of S. pombe DNA has been constructed in the plasmid vector, pDB248', which is capable of high frequency transformation of S. pombe. Procedures are described which enable plasmids to be recovered from S. pombe by their reintroduction into E. coli. These methods have been used to detect the S. pombe genes lys 1+, ade 6+ and his 2+ in the gene bank by complementation of mutant gene functions, and to physically isolate the lys 1+ gene. PMID- 6294467 TI - In vitro construction of the tufB-lacZ fusion: analysis of the regulatory mechanism of tufB promoter. AB - To investigate the regulatory mechanism of the tufB operon, we have constructed plasmids in which the lac structural genes have been fused to the regulatory region and the 5'-coding sequence of the tufB gene. The fusion was performed by incorporating the 6.6 kb EcoRI-HpaI fragment of plasmid pTUB1, which carried the tufB gene (Miyajima et al. 1979), into the EcoRI and SmaI sites of pMC1403 lac fusion vector (Casadaban et al. 1980). This gene fusion resulted in the production of a hybrid protein consisting of the N-terminal portion (12 amino acid residues) of EF-TuB and the enzymatically active C-terminal half of beta galactosidase. Bacteria harboring the recombinant plasmid showed a strong Lac+ phenotype. In such a fusion, the lac gene expression was under the control of the tufB promoter. This was evidenced by the following observations; (i) the tufB lacZ hybrid protein was synthesized constitutively; (ii) its production augmented in parallel with the increase in growth rate; and (iii) on carbon-source upshift, the hybrid protein was produced at a rate 2.5-fold higher than that of the mass increase. Several derivatives of this recombinant plasmid harboring deletions and/or inversions in the tufB regulatory region have been constructed and their properties are described. PMID- 6294468 TI - Origin of replication of Escherichia coli plasmid RSF 1030. AB - The nucleotide sequence of a region of plasmid RSF 1030 that includes the origin of DNA replication was determined using the DNA of a small derivative, pST19. The nucleotide sequence of the pST 19 origin region is very similar to that of the ColE1 origin except for a 25 base pair (bp) deletion about 350 bp upstream of the origin and a considerable difference in the region between 400 and 600 bp upstream of the origin. Replication of pST19 starts at one of three consecutive nucleotides (dA, dA or dC) located at a unique position in the region where the nucleotide sequence is identical to that of the ColE1 origin. There are two major sites of initiation of transcription in the region. Transcription from one of the sites yields the primer precursor that can be cleaved by RNase H to form the primer of about 530 nucleotides long. Transcription from the other site proceeds on the opposite strand and terminates close to the primer initiation site to yield species I RNA (or RNA I) about 105 nucleotides long. The presumed RNA polymerase binding sites in the promoters of these transcripts differ from those of the corresponding ColE1 transcripts. Incompatibility specified by pST19 is different from that specified by ColE1. Hypothetical peptides encoded by the origin region of these plasmids are unlikely to be involved in the determination of incompatibility. It has been shown that RNA I is an incompatibility-group specific inhibitor of primer formation. Despite a significant difference in nucleotide sequence, the primer RNA and RNA I of pST19 can be folded into structures analogous to those of the ColE1 transcripts. PMID- 6294469 TI - DNA-mediated gene transfer in Chinese hamster ovary cells: clonal variation in transfer efficiency. AB - Thymidine kinase-deficient Chinese hamster ovary (CHO) cells were genetically transformed with the BamHI restriction fragment encoding the thymidine kinase gene of herpes simplex virus (HSV-tk). We have observed considerable clonal variation among independent CHO sublines with respect to transformation competence for the DNA-mediated gene transfer of HSV-tk. Transformation frequencies greater than or equal to 3 X 10(-4) were observed consistently in one subline, with a transformation efficiency of approximately 1 transformant per ng viral gene. The frequency and efficiency of transformation we observed in this system are at least 10-fold greater than those previously reported for DNA mediated transformation of CHO cells by HSV-tk. All of the CHO HSV-tk+ transformants examined were stable for the transferred genotype in the absence of selection, and all showed evidence of co-transformation by unselected plasmid pBR322 sequences. PMID- 6294471 TI - Localization of the metJBLF gene cluster of Escherichia coli in lambda met transducing phage. AB - The position of the metJBLF gene cluster in the transducing phage lambda met102 was determined by ligation of its leftmost EcoRI fragment (102-1) to the lambda BCDEF (nin5) EcoRI fragment of lambda gtl (lambda BC) and characterization of the resultant recombinant phage. The new transducing phage carries about 6kb of bacterial DNA which contains the entire met gene cluster including the promoter of its rightmost member metF. Reasonable estimates of the coding capacity required for the four genes indicate that most of the bacterial DNA of the recombinant phage is occupied by the met gene cluster. PMID- 6294470 TI - The organization and regulation of the pyrBI operon in E. coli includes a rho independent attenuator sequence. AB - 1. The two polypeptide chains that comprise aspartate carbamoyltransferase in Escherichia coli are encoded by adjacent cistrons expressed in the order, promoter-leader-catalytic cistron-regulatory cistron (p-leader-pyrBI). These two cistrons and their single control region have been cloned as a 2,800 base pair (bp) fragment (The minimal coding requirement for the catalytic and regulatory polypeptides is about 1,350 bp plus control regions). The genes contained by this fragment are subject to normal repression controls and thus possess the intact control regions. 2. By deleting an internal fragment with specific restriction endonucleases, it was possible to construct shortened fragments which no longer produced the regulatory polypeptide. In these cases the expression of the catalytic cistron was normal and subject to repression upon growth in the presence of uracil. Since the pyrB cistron retained transcriptional control, the regulatory polypeptide was not required for expression or control of the catalytic cistron. As expected, the catalytic trimer (Mr = 100,000 daltons) from these deletion mutants had no effector response nor did it exhibit homotropic kinetics for aspartate. The enzyme was identical to the c3 trimer purified from the native holoenzyme by neohydrin dissociation. 3. Insertion of Mu d1(lac Apr) into the structural region of pyrB had a negative effect on the expression of pyrI. This supports the idea that the catalytic and regulatory polypeptide chains of aspartate carbamoyl-transferase are encoded by a single bicistronic operon. Detailed restriction analysis of the cloned pyrBI region has produced a genetic map of restriction sites which is colinear with the published amino acid sequences of the two polypeptides. These maps indicate that the 3'-terminus of the catalytic cistron is adjacent to the 5'-terminus of the regulatory cistron and separated by 10-20 bp. 4. DNA sequence analysis of the 5'-proximal regions of pyrBI revealed that an extensive leader sequence separated the promoter and first structural gene pyrB. This leader of approximately 150 bp contains an attenuator sequence and the translational signals required for the production of a leader polypeptide of 43 amino acids. In this paper we describe the structural organization of pyrBI, and provide a detailed analysis of its regulatory region including its DNA sequence. PMID- 6294472 TI - Mutants that overproduce TraTp, a plasmid-specified major outer membrane protein of Escherichia coli. AB - The isolation of a series of plasmid mutant derivatives that overproduce the traT outer membrane protein, TraTp, is described. Some of the mutants directed the synthesis of 10-fold more TraTp (200,000 copies/cell) than did the parental plasmid (20,000 copies/cell). The proteins specified by all mutant plasmids except one were correctly inserted into the outer membrane and exposed on the cell surface. The TraTp that was not correctly inserted did not mediate the expected levels of surface exclusion and serum resistance, suggesting that surface localization is a requirement of TraTp function. The overproduction of TraTp was deleterious to bacterial growth, particularly that of minicell mutants of E. coli K-12. PMID- 6294473 TI - The role of the transforming A gene of SV40 in the mutagenic activity of the virus. AB - The mutagenic activity of the tsA239 mutant of SV40 which synthetizes a defective T antigen at 40 degrees C was investigated in Chinese hamster cells under permissive and nonpermissive temperature. At 33 degrees C the virus increased the yield of 6-mercaptopurine-resistant colonies after 2 days expression time by a factor of 1.6-4 as compared with the control and raised the frequency of aberrant metaphases after the same time by a factor of 1.9-3.4. In the same experiments, with the same initially infected population of Chinese hamster cells, at 40 degrees C tsA SV40 did not induce either gene mutations or chromosome aberrations at the same early stage after infection. Presumably the activity of the A gene of SV40 is necessary not only for the transforming but also for the mutagenic effect of the virus. PMID- 6294474 TI - The sites of action of the two copy number control functions of plasmid R1. AB - Two negatively acting functions - the CopA-RNA and the CopB protein - are involved in the control of replication of plasmid R1. They both act as inhibitors of expression of a gene, repA, which seems to be positively required for autonomous plasmid replication. Here we show that the two control functions act separately and independently. The CopB protein represses initiation of transcription of the repA gene, and its target site lies within a 60 base pair region containing the repA promoter. The CopA-RNA acts downstream of the repA promoter in the leader sequence containing the copA gene itself, preceding the repA structural gene. Measurements of RepA-beta-galactosidase expression from wild-type and a copA mutant fusion hybrid in the presence of extra copies of the respective copA genes show that a point mutation affecting the activity of the CopA-RNA can also affect CopA target properties. It is therefore concluded that the target site for the CopA-RNA resides within the copA gene in a small region encoding the loop of a stem-loop structure in the CopA-RNA. In addition, the data indicate a direct nucleic acid-nucleic acid interaction as the basis for the CopA inhibitor activity. PMID- 6294475 TI - Cloning of bacterial DNA replication genes in bacteriophage lambda. AB - Recombinant lambda phages containing the genes for dnaZ protein (the gamma subunit of DNA polymerse III holoenzyme), primase (dnaG protein) and dnaC protein from Escherichia coli and Salmonella typhimurium were isolated. Each gene cloned from S. typhimurium has extensive DNA sequence homology to the corresponding E. coli gene. Clones selected by complementation of a dnaA temperature-sensitive mutant appear similar to other isolated suppressors of dnaA (Projan and Wechsler 1981). Derivatives of each cloned fragment suitable for overproduction of the protein were constructed. Of those tested, only the phage containing the E. coli dnaZ gene resulted in significant overproduction. PMID- 6294476 TI - DNA sequence analysis of a Drosophila foldback transposable element rearrangement. AB - The complete nucleotide sequence of a DNA rearrangement associated with the foldback 4 (FB 4) transposable element is presented. The results demonstrate that the entire loop sequence and almost all of one of the inverted terminal repeats is absent. Moreover, the sequence of the remaining inverted repeat suggests that the FB elements might undergo inversions via recombinations between the two inverted repeats of a single element. PMID- 6294477 TI - Repair of psoralen-induced crosslinks in cells multiply infected with SV40. AB - Experiments were conducted to study the relationship between the production of interstrand crosslinks by 4,5',8-trimethylpsoralen (psoralen) in simian virus 40 DNA and the ability of psoralen to inactivate the virus. Under conditions where only single viral particles enter a given host cell, approximately one crosslink was lethal to the virus and could not be repaired. In contrast, when multiple viral genomes infected a host cell, psoralen-induced crosslinks were repaired (multiplicity reactivation). A model is proposed for multiplicity reactivation which involves genetic recombination between damaged viral genomes. PMID- 6294478 TI - Marker rescue allows direct selection for recombinant plasmids in streptococci. AB - Resident deletion derivatives (Ems or Cms) of the streptococcal plasmid vector pGB301 rescue antibiotic resistance genes from linearized pGB301 (Emr, Cmr) DNA with high frequency. Insertion of passenger DNA next to an antibiotic resistance determinant of pGB301, which is missing on the resident plasmid, forces corescue of these two plasmid domains, thus allowing direct selection for recombinant plasmids. PMID- 6294479 TI - The Red function of phage lambda mediates the alteration of an interspersed repeated DNA sequence from the kangaroo rat Dipodomys ordii. AB - The kangaroo rat D. ordii is shown to contain an interspersed repeated sequence 3.25 kb in length. This sequence undergoes alteration (deletions) when cloned into a Red+ lambda vector but is stable in a Red- vector. Using the Red- clone it is shown that deletions are the result of the activity of the lambda Red, or E. coli RecE, recombination systems. Mixed infection experiments demonstrate that this activity is acting in trans. The deletions appear to yield fragments of specific size, suggesting that the recombination system is recognizing a specific sequence. The demonstration of small homologous fragments in the animal genomic DNA suggests that a similar system of recombination may exist in D. ordii. PMID- 6294480 TI - Insertion of nonhomologous DNA into the yeast genome mediated by homologous recombination with a cotransforming plasmid. AB - Bacterial plasmids containing no detectable homology with yeast DNA sequences were inserted into the yeast genome by cotransforming with a plasmid containing a yeast gene. Analysis of the yeast transformants confirmed that recombination events occurred between the prokaryotic sequences shared by the two plasmids and between the yeast sequences common to the cotransforming plasmid and to the genome. Multiple copies of the two plasmids, in both tandem and interspersed arrays, are inserted by this method. Populations of cells grown from individual transformants are heterogeneous for the number of integrated sequences. The number of integrated bacterial sequences is greatly reduced after 100 generations of growth in the populations that initially contained large numbers of sequences, while it is stable in those populations that initially contained either a single or a small number of copies. PMID- 6294481 TI - The cytochrome oxidase subunit I split gene in Saccharomyces cerevisiae: genetic and physical studies of the mtDNA segment encompassing the 'cytochrome b homologous' intron. AB - We have constructed a refined genetic and physical map of 38 oxi3 mutations. With the help of the rho- clones derived from 'short' and 'long' genes, pairwise crosses between mutants, estimations of their reversion frequencies and analyses of mitochondrially synthesized proteins, we have characterized and localized several mutants in the exon A4 and in the intron aI4. We present genetic and physical evidence that in the 'long' gene the exon A5 is split into at least three quite distinct exons, A5-1, A5-2 and A5-3 where numerous mutations are localized. We suggest that a novel 56 Kd polypeptide, which accumulates in some cis-dominant oxi3- mutants results from the translation of the upstream exons and the downstream aI4 intron. PMID- 6294482 TI - On the role of the single-stranded DNA binding protein of bacteriophage T4 in DNA metabolism. I. Isolation and genetic characterization of new mutations in gene 32 of bacteriophage T4. AB - The product of gene 32 of bacteriophage T4 is a single-stranded DNA binding protein involved in T4 DNA replication, recombination and repair. Functionally differentiated regions of the gene 32 protein have been described by protein chemistry. As a preliminary step in a genetic dissection of these functional domains, we have isolated a large number of missense mutants of gene 32. Mutant isolation was facilitated by directed mutagenesis and a mutant bacterial host which is unusually restrictive for missense mutations in gene 32. We have isolated over 100 mutants and identified 22 mutational sites. A physical map of these sites has been constructed and has shown that mutations are clustered within gene 32. The possible functional significance of this clustering is considered. PMID- 6294483 TI - DNA replication and transcription in a temperature-sensitive mutant of E. coli with a defective DNA gyrase B subunit. AB - A temperature-sensitive mutant of E. coli with a defective DNA gyrase B subunit has been obtained. The mutation is expressed in the thermolability of DNA gyrase in vitro and in DNA relaxation in vivo. DNA replication in the mutant does not stop under non-permissive conditions; its rate gradually falls by a factor of 2 to 3. The transcription rate also drops by a factor of 2 to 3, but before replication. Small concentrations of rifampicin, an inhibitor of bacterial RNA polymerase, make for a partial survival of the mutant cells under non-permissive conditions. The results suggest the conclusion that DNA supercoiling is mainly required to ensure the optimum transcription level in the cell. PMID- 6294484 TI - Glycerol-releasing activity of histamine-sensitizing factor of Bordetella pertussis for rat adipocytes in vitro. AB - Histamine-sensitizing factor (HSF) purified from Bordetella pertussis induced specifically the release of glycerol from rat epididymal adipocytes in vitro. The most sensitive and reproducible results were obtained by using 1 to 2 x 10(5) adipocytes/tube from rats weighing 150 to 200 g, and by incubation at 37 C for 180 min. After a lag period of about 60 min, HSF-treated adipocytes released glycerol in increasing amounts between 60 and 240 min, depending on the dose of HSF. A close correlation between the glycerol-releasing (GR) activity of HSF for adipocytes and histamine-sensitizing or leukocytosis-promoting activity in mice was observed. The GR activity was inactivated by heating at 56 C for 60 min, 63 C for 30 min or 96 C for 10 min. The adipocytes washed out with a Krebs-Ringer bicarbonate buffer immediately after being exposed to HSF for 1 to 3 min manifested about 75% of the total GR activity induced by HSF, and those washed out after being exposed for 30 min or longer had full activity. Anti-HSF serum neutralized the activity when it was added to adipocytes simultaneously with HSF, but did not when it was added 30 min after being exposed to HSF. By using both native and 125 I-labeled HSF, the ratio of binding of HSF to adipocytes was estimated to be 10 to 15% of the total HSF per 2 x 10(5) cells/tube, and to be about 1,000 molecules of HSF per cell to induce the release of glycerol. The GR activity induced with 10 ng of HSF was inhibited by addition of insulin at a dose of over 1 micro IU/tube, but not by concanavalin A. PMID- 6294485 TI - Effect of interferon on the production of HBsAg and induction of an antiviral state in human hepatoma cell line PLC/PRF/5. AB - The effects of human alpha and beta interferons (IFN) on the production of HBsAG by PLC/PRF/5 cells, an HBsAg-producing human hepatoma cell line, were studied in the exponential and stationary phases of cell growth. When exponential phase cells were treated with 100 or 1,000 U of IFN per ml for 48 hr. the amount of HBsAg in the culture medium decreased. The number of cells and the synthesis of DNA and proteins were also reduced by the IFN treatment. These results suggested that IFN did not affect the production of HBsAg specifically in exponential phase cells. When cells in the stationary phase were similarly treated with IFN, HbsAg production was not inhibited nor did the number of cells decrease. To examine the antiviral state induced by IFN in PLC/PRF/5, induction of 2'5'-oligo (A) synthetase and susceptibility to two kinds of viruses were examined. The 2'5' oligo (A) synthetase activity was increased in an IFN-dose dependent manner. Susceptibility to vesicular stomatitis virus (VSV) and encephalomyocarditis virus (EMCV) was decreased by treatment with 10 and 100 U of IFN per ml for 20 hr. It was concluded that IFN-alpha and IFN-beta induce 2'5'-oligo (A) synthetase and the antiviral state, but do not inhibit HBsAg production by PLC/PRF/5 cells. PMID- 6294486 TI - New host specificity of DNA produced by Escherichia coli carrying trimethoprim resistant R plasmid. PMID- 6294487 TI - Temperature dependence of bactericidal action of polymyxin B. PMID- 6294488 TI - Detection of human rotavirus by reversed passive hemagglutination (RPHA) using antibody against a cultivable human rotavirus as compared with electron microscopy (EM) and enzyme-linked immunosorbent assay (ELISA). PMID- 6294489 TI - Efficacy of intradermal administration of herpes simplex virus subunit vaccine. PMID- 6294490 TI - Lacrimal gland carcinoma. PMID- 6294492 TI - Late recurrence of Wilms tumor. AB - Three patients with Wilms tumors recurring after nine years in an organized abdominal hematoma, after ten years in the lung, and after eleven years in the central nervous system, respectively, are presented. It is proposed that the nephroblastoma cells have survived in a latent or a very slowly growing state. The reactivation mechanisms are briefly discussed. A review of the literature indicates a rate of recurrence of nonmetastasizing Wilms tumors later than 24 months of 1.4--3.9%. PMID- 6294491 TI - [Carcinogenesis of mineral fibers: state of the art]. PMID- 6294493 TI - A differentiation-dependent polyisoprenol kinase in Dictyostelium discoideum. AB - Crude membrane fractions of Dictyostelium discoideum show the capacity to synthesize (1--3H)dolicholphosphate from (1--3H)dolichol. Formation of dolicholphosphate increased continuously over the first 15 min. The reaction rate was nearly linear with respect to the dolichol content up to 150 microM. The phosphate donor for the reaction is CTP. The optimum concentration of CTP is about 0,75 mM. The reaction is dependent on divalent metal ions, magnesium being more effective than calcium or manganese. The activity of the polyisoprenol kinase depends on the course of the early development. Maximum enzyme activities are present 4--6 h after the induction of the differentiation. PMID- 6294495 TI - Thialysine and selenalysine as allosteric inhibitors of E. coli aspartokinase III. PMID- 6294494 TI - Liver phosphorylase phosphatase. PMID- 6294496 TI - Positive correlation between superoxide release and intracellular adenosine deaminase activity during macrophage membrane perturbation regardless of nature or magnitude of stimulus. PMID- 6294497 TI - Effect of the regulatory subunit of cAMP-dependent protein kinase on the genetic activity of eukaryotic cells. PMID- 6294498 TI - Protein synthesis in lysates of Aedes albopictus cells infected with vesicular stomatitis virus. AB - Aedes albopictus cells (clone LT-C7) showed a marked cytopathic effect and inhibition of protein synthesis (both host and viral) after infection with vesicular stomatitis virus (VSV), but only if (i) cultures were incubated at 34 degrees C rather than 28 degrees C and (ii) serum was present in the medium (S. Gillies and V. Stollar, Mol. Cell. Biol. 2:66-75, 1982). To learn more about how protein synthesis is shut off in VSV-infected A. albopictus cells, we have compared cell-free protein synthesis in extracts prepared from VSV-infected cells and control cells. Extracts prepared 6 h after infection from VSV-infected cells maintained at 34 degrees C in the presence of serum reflected what was observed with intact cells in at least two respects: (i) they showed a markedly diminished capacity to carry out protein synthesis (whether directed by endogenous or exogenously added mRNA), and (ii) there was decreased phosphorylation in vitro by [gamma-32P]ATP of a specific ribosomal protein (Gillies and Stollar, Mol. Cell. Biol. 2:66-75, 1982). In addition, and consistent with a block at the level of initiation, the formation of 80S initiation complexes, as measured by binding of VSV 12 to 18S mRNA, was reduced in the inactive extracts. Addition of an S-100 fraction from uninfected cells to the inactive extract reversed each of the aforementioned changes; i.e., it restored protein synthetic activity, it stimulated the formation of 80S initiation complexes, and it increased phosphorylation of the specific ribosomal protein referred to above. The active component in the S-100 fraction was heat labile and non-dialyzable and, upon ammonium sulfate fractionation of the S-100 fraction, was found in the 40 to 70% saturation fraction. Our findings suggest that VSV infection of A. albopictus cells inhibits protein synthesis by inactivating a macromolecular component, probably a protein, in the S-100 fraction which may be involved in the initiation of protein synthesis. More specifically, we suggest that this component is involved in the joining of the ribosomal subunits to form 80S initiation complexes. PMID- 6294499 TI - Revertants of an S49 cell mutant that expresses altered cyclic AMP-dependent protein kinase. AB - Dibutyryl adenosine 3',5'-phosphate (Bt2cAMP)-sensitive (Bt2cAMPS) revertants were isolated from a resistant S49 cell mutant carrying a structural gene lesion in the regulatory subunit of cAMP-dependent protein kinase (cA-PK). This was accomplished with a counter-selection in which, first, Bt2cAMP was used to reversibly arrest revertants, and then a sequence of treatments with bromodeoxyuridine, 33258 Hoechst dye, and white light was used to kill cycling mutant cells. Reversion rates in nonmutagenized cultures could not be accurately measured, but spontaneous revertants do occur and with frequencies of less than 10(-7) to 10(-5). The mutagens ethyl methane sulfonate (EMS), N-methyl-N'-nitro-N nitro-soguanidine (MNNG), and ICR191 increased the reversion frequency. In all cases, reversion to Bt2cAMP sensitivity was associated with restoration of wild type levels and apparent activation constant for cAMP of cA-PK. MNNG induced revertants whose cell extracts contained cA-PK activity distinguishable from that of wild type by thermal liability. EMS did not. The counter-selection effectively isolates rare phenotypes and is therefore a useful tool in further somatic genetic experiments. The association of reversion with alterations in cA-PK function supports all previous data from this and other laboratories implicating cA-PK as the intracellular mediator of cAMP effects. Reversion is probably the result of a mutational event. Induction of reversion by ICR191 suggests the existence of a novel mechanism for generating revertants in somatic cells. PMID- 6294500 TI - Deletions of N-terminal sequences of polyoma virus T-antigens reduce but do not abolish transformation of rat fibroblasts. AB - Polyoma virus transforms, upon infection or DNA transfection, nonpermissive Fisher rat fibroblasts. Cloned viral DNA was deleted of sequences around the Bg/I site at nucleotide 86 by Bal31 nuclease treatment and then recloned in Escherichia coli. The extent of deletion for each mutant was then determined by DNA sequencing. Deletions included the early transcription control signals; others stretched into the N-terminal coding sequences of the viral tumor antigens. The transformation efficiency of 16 mutants was tested by transfecting rat fibroblasts. Expression of the T antigens was analyzed by immunofluorescence detection after transfection of rat fibroblasts, mouse secondary embryo cells, and HeLa cells. We found that the absence of the early transcription control sequences (TATA and CAAT boxes) did not significantly alter the transformation capacity of the virus. On the other hand, deletion of the initiator methionine ATG codon or further into the coding sequences did abolish the transformation capacity in some mutants, whereas others maintained a reduced transforming activity, possibly by initiation of translation in a penultimate methionine. PMID- 6294501 TI - Polyacrylamide gel mapping of chicken tRNA: comparison of polysome-bound and whole-cell tRNA from normal and avian sarcoma virus-infected chicken embryo fibroblasts. AB - Analysis of the tRNA population from chicken cells was performed by means of polyacrylamide gel mapping. About 60 species were detected; most of these were positively identified by their acceptor specificity. The comparison of polysome bound and overall cellular tRNA gel patterns from normal and Rous sarcoma virus infected chicken embryo fibroblasts led us to the following observations: some tRNA species were present in the same relative proportions in all the preparations, and within isoaccepting groups the same species was preponderant; however, although about 8% of whole-cell tRNA was recovered in polysomal preparations, amounts ranging from 3 to 30% were found for individual tRNA species. This points to the absence of a direct correlation between the amount of each mature tRNA species produced and the frequency with which it is used in this case of embryonic cells. No significant difference was observed between the whole cell tRNA patterns from normal and infected cells. Thus, tRNA transcription appears unaltered when cells are transformed and virus producing. No change was observed in the extent of a post-transcriptional modification of tRNAPhe (the base Y). However, viral infection led to some changes in the relative proportions of individual species from polysomal preparations. PMID- 6294502 TI - Somatic cells efficiently join unrelated DNA segments end-to-end. AB - Molecular substrates for probing nonhomologous recombination in somatic cells were constructed by inserting pBR322 sequences at selected sites on the simian virus 40 (SV40) genome. The chimeric products are too large to be packaged into an SV40 capsid. Therefore, production of viable progeny requires that most of the pBR322 sequences be deleted without altering any SV40 sequences that are essential for lytic infection. As judged by plaque assay, these recombination events occur at readily detectable frequencies after transfection into CV1 monkey kidney cells. Depending on the site of pBR322 insertion, the infectivities of the full-length circular or linear chimeras ranged from 0.02 to 2% of the infectivity of linear wild-type SV40 DNA. Nucleotide sequence analysis of several recombinant progeny revealed three distinct classes of recombination junction and indicated that the causative recombination events were minimally dependent on sequence homology. Potential mechanisms involving recombination at internal sites or at ends were distinguished by measuring the infectivity of chimeric molecules from which various lengths of pBR322 had been removed. These data support end-to-end joining as the primary mechanism by which DNA segments recombine nonhomologously in somatic cells. This end joining appears to be very efficient, since SV40 genomes with complementary single-stranded tails or with short non-complementary pBR322 tails were comparably infectious. Overall, this study indicates that mammalian somatic cells are quite efficient at the willy-nilly end-to-end joining of unrelated DNA segments. PMID- 6294503 TI - Genetic selection for reciprocal translocation at chosen chromosomal sites in Saccharomyces cerevisiae. AB - We have constructed viable Saccharomyces cerevisiae strains containing a reciprocal translocation between the URA2 site of chromosome X and the HIS3 site of chromosome XV. Our methodology is an extension of the method originally developed to introduce an altered cloned sequence at the chromosomal location from which the parent sequence was derived (S. Scherer and R.W. Davis, Proc. Natl. Acad. Sci. U.S.A. 76:4951-4955, 1979). It comprises three essential steps. First, a nonreverting ura2- strain was constructed by deleting a 3.7-kilobase fragment from the coding sequence of the wild-type URA2 gene. Second, part of the coding sequence of the wild-type URA2 gene (without promotor) was inserted at the HIS3 locus of the ura2- strain. Third, after several generations of growth on uracil-supplemented medium, ura2+ colonies were selected which resulted from mitotic recombination between the nonoverlapping deletions of URA2 located on chromosomes X and XV. PMID- 6294504 TI - Cyclic AMP-modulated phosphorylation of intermediate filament proteins in cultured avian myogenic cells. AB - The intermediate filament proteins desmin and vimentin and the muscle tropomyosins were the major protein phosphate acceptors in 8-day-old myotubes incubated for 4 h in medium containing radiolabeled phosphate. The addition of isoproterenol or 8-bromo-cyclic AMP (BrcAMP) resulted in a two- to threefold increase in incorporation of 32PO4 into both desmin and vimentin, whereas no changes in the incorporation of 32PO4 into tropomyosin or other cellular proteins were observed. The BrcAMP- or hormonally induced increase in 32PO4 incorporation into desmin and vimentin was independent of protein synthesis and was not caused by stimulation of protein phosphate turnover. In addition, BrcAMP did not induce significant changes in the specific activity of the cellular ATP pool. These data suggest that the observed increase in 32PO4 incorporation represented an actual increase in phosphorylation of the intermediate filament proteins desmin and vimentin. Two-dimensional tryptic analysis of desmin from 8-day-old myotubes revealed five phosphopeptides of which two showed a 7- to 10-fold increase in 32PO4 incorporation in BrcAMP-treated myotubes. Four of the phosphopeptides identified in desmin labeled in vivo were also observed in desmin phosphorylated in vitro by bovine heart cAMP-dependent protein kinase. Although phosphorylation of desmin and vimentin was apparent in myogenic cells at all stages of differentiation, BrcAMP- and isoproterenol-induced increases in phosphorylation of these proteins were restricted to mature myotubes. These data strongly suggest that in vivo phosphorylation of the intermediate filament proteins desmin and vimentin is catalyzed by the cAMP-dependent protein kinases and that such phosphorylation may be regulated during muscle differentiation. PMID- 6294506 TI - Murine cell complementation of a cold-sensitive defect for simian virus 40 replication in simian cells. AB - Mouse 3T3 fibroblasts were found to complement, in simian cell variants semipermissive to simian virus 40, a cold-sensitive defect of an early function, but not a nonconditional defect of viral uncoating. The variant simian cells could rescue simian virus 40 from 3T3 transformants, and this capacity was not temperature dependent. PMID- 6294505 TI - Gene transfer: DNA microinjection compared with DNA transfection with a very high efficiency. AB - We have developed a procedure that gives a very high efficiency of transfection in mammalian cells with low-molecular-weight DNA (approximately 10(4) base pairs). The procedure uses cells in suspension that are shocked with polyethylene glycol 4 h after replating. We compared this transfection technique to the standard technique involving manual microinjection of DNA into the nuclei of mammalian cells, using recombinant plasmids containing the simian virus 40 A gene or the herpes simplex virus thymidine kinase gene or both. The efficiency of transfection depends on a number of variables, the most important of which is the difference in transfectability of different cell lines. In our laboratory, the cell line that had the highest efficiency of transfection was tk-ts13, which is derived from baby hamster kidney cells that are deficient in thymidine kinase and temperature sensitive for growth. Under the appropriate conditions, as many as 70% of these cells can be transfected so that transient gene expression can be detected. With the manual microinjection technique, gene expression is independent of the cell line used and occurs faster than after transfection. The results suggest that the critical stage in transfection is the delivery of DNA molecules to the nucleus. Our experiments also indicate that an enzymatic function, in our case, thymidine kinase activity, gives a higher percentage of positive transfectants than when proteins are visualized only by indirect immunofluorescence. The transfection procedure described in this paper is simple and reproducible and, although less efficient than microinjection, ought to be useful in phenotypic and genotypic studies in which transfer of genes to a large number of cells is desirable. PMID- 6294508 TI - Complement receptor enhancement by chemotactic factors. PMID- 6294507 TI - Membrane perturbation and stimulation of arachidonic acid metabolism. PMID- 6294509 TI - [The sublingual and submandibular glands after preganglionic parasympathetic blockade. A histological, electron optical, autoradiographic and scintigraphic study on the guinea pig]. AB - After preganglionic parasympathetic block three histological and histochemical phases could be determined in the only cholinergically innervated mucous cells of the sublingual gland. The first phase is characterized by overproduction and oversecretion. Possibly this results from acetylcholine overproduction, caused by the block, termed as "degeneration secretion" however of preganglionic origin. Hypersensitivity of the gland parenchyma appears to be present also, which with catecholamins as well as acetylcholine cause the ensuing "paralytic secretion". At the end of the first phase the desquamation of parts of cells, and rarely entire cells, in form of apocrine and holocrine secretion occurs and a uniform cell population is developing. When this cell population develops the second phase starts. The lack of stimulation because of acetylcholine exhaustion is discussed as the cause of the second phase. In the third phase a self regulatory involution commences. In the double innervated serous submandibular gland the effect of chorda tympani section was less severe. PMID- 6294510 TI - Pharmacology and clinical use of ranitidine. PMID- 6294511 TI - [Infectious mononucleosis]. PMID- 6294512 TI - [Cushing syndrome]. PMID- 6294513 TI - [African histoplasmosis; immunological parameters and treatment with ketoconazole]. PMID- 6294514 TI - Enhanced differentiation of isolates of Candida albicans using a modified resistogram method. PMID- 6294515 TI - Subcellular distribution of adenylate cyclase, cyclic-AMP phosphodiesterase, protein kinases and phosphoprotein phosphatase in Trypanosoma brucei. AB - The subcellular distribution of adenylate cyclase, cyclic-AMP phosphodiesterase, protein kinases and phosphoprotein phosphatase in bloodstream forms of Trypanosoma brucei was determined by isopycnic sucrose-gradient centrifugation of post-large-granule extracts. Cyclic-AMP phosphodiesterase was almost entirely soluble whereas adenylate cyclase was membrane-bound. The latter enzyme appeared to be absent from the plasma-membrane fraction but copurified with acid phosphatase and acid phosphodiesterase indicating a possible association with the flagellar pocket. At least two protein kinase activities could be distinguished as based on their distribution profiles in gradients, their preference for exogenously added acceptor protein and their inhibition and stimulation by suramin and nucleoside, respectively. Suramin-sensitive protein kinase co purified with the plasma-membrane marker alpha-D-glucosidase and a nucleoside stimulated protein kinase behaved as a typical cell-sap enzyme. Phosphoprotein phosphatase activity was found to be mainly soluble but a small part seemed to be associated with plasma membranes. PMID- 6294516 TI - MMWR. PMID- 6294517 TI - Intensive immunosuppression in progressive multiple sclerosis. A randomized, three-arm study of high-dose intravenous cyclophosphamide, plasma exchange, and ACTH. AB - Fifty-eight patients with severe, progressive multiple sclerosis were prospectively randomized to one of three treatments: 20 received intravenous ACTH, 20 received high-dose intravenous cyclophosphamide plus ACTH, and 18 were placed on a regimen consisting of plasma exchange, low-dose oral cyclophosphamide, and ACTH. The three groups were similar in age, sex, duration and type of disease, and degree of disability. Before treatment and six months and one year after treatment, a disability-status score, ambulation index, and functional-status score were determined, and a quantitative neurologic examination was performed. In the ACTH group, the number of patients stabilized or improved was 8 of 20 at six months and 4 of 20 at one year; in the cyclophosphamide-ACTH group, 18 of 20 at six months and 16 of 20 at one year; and in the plasma exchange group, 11 of 18 at six months and 9 of 18 at one year. High-dose cyclophosphamide plus ACTH was most effective in halting progression of the disease at both 6 and 12 months (at 12 months, cyclophosphamide-ACTH vs. ACTH, P = 0.0004; cyclophosphamide-ACTH vs. plasma exchange, P = 0.087). Thus, progressive multiple sclerosis may be stabilized by short-term, intensive immunosuppression with cyclophosphamide plus ACTH. PMID- 6294518 TI - Unstable methotrexate resistance in human small-cell carcinoma associated with double minute chromosomes. PMID- 6294519 TI - Treatment of multiple sclerosis. PMID- 6294520 TI - Elevated angiotensin I-converting enzyme in histiocytic medullary reticulosis. PMID- 6294521 TI - Stability of delta-9-tetrahydrocannabinol in stored blood and serum. PMID- 6294522 TI - The constituents of cannabis and the disposition and metabolism of cannabinoids. PMID- 6294523 TI - Radioimmunoassays for cannabinoids. AB - The simplicity, sensitivity, and specificity of radioimmunoassay have made it an attractive procedure for the analysis of delta-9-tetrahydrocannabinol (THC) in biological fluids or tissues. The presence of closely related compounds such as metabolites may interfere with radioimmunoassay results. Appropriate design of immunogens may diminish such interference. This work has been directed towards the use of the amyl side chain for linking cannabinoid compounds to proteins to form immunogens. Although the amyl side chain is metabolized to some extent, the metabolites are not quantitatively significant in most cases. 5'-Carboxy-delta-8 THC and 5'-carboxy-delta-9-THC were linked to bovine serum albumin. Immunization of rabbits with the resulting conjugates resulted in the formation of antisera with high selectivity for delta-9-THC vs. its carboxylic acid metabolite, 11-nor 9-carboxy-delta-9-THC. Delta-8-THC radioligands (4',5'-tritium and 5'-iodine-125) could be used with these antisera for analysis of delta-9-THC in plasma. Sensitivity with tritium-labeled material is about 2.5 ng/ml. 5'-Oxo-11-nor-9 carboxy-delta-8-THC was used to prepare an immunogen which led to the generation of an antiserum highly specific for 11-nor-9-carboxy-delta-9-THC. This antiserum and iodine-125-5'-iodo-11-nor-9-carboxy-delta-8-THC were used to develop a highly specific assay for 11-nor-9-carboxy-delta-9-THC in plasma. PMID- 6294524 TI - Radioimmunoanalysis of delta-9-THC in blood by means of an 125I tracer. AB - A radioimmunoassay for delta-9-THC in plasma, whole blood, or hemolyzed blood specimens has been presented. Samples and standards were diluted with methanol and centrifuged. An aliquot of the supernatant fluid was incubated with RIA buffer, 125I-labeled delta-8-THC and rabbit anti-THC serum. Solid phase goat anti rabbit immunoglobulins were added to separate bound from free THC. After centrifugation the supernatant fluid was aspirated and the radioactivity of the precipitate was counted in a gamma counter. The concentration of THC was calculated from a standard curve using the logit-log transformation of the average counts of duplicate tubes. The assay had several advantages. Methanol dilution gave better results than direct analysis. The 125I-labeled THC had high specific activity and could be counted in a gamma counter. The immunological separation of antibody-bound THC from free THC was better than separation techniques using ammonium sulfate and activated charcoal. THC was determined in 0.1 ml of sample with a sensitivity of 1.5 ng/ml in plasma and 3.0 ng/ml in hemolyzed blood. PMID- 6294525 TI - Significant developments in radioimmune methods applied to delta 9-THC and its 9 substituted metabolites. AB - Novel RIA's have been developed for marijuana detection. Modified simple techniques measure the "acute" metabolites in minute saliva, breath, and dried blood samples. Solid purified antibody matches the sensitivity and yields sensitivity values identical to liquid assays. The importance of the polar 9 substituted metabolites (9SM) of THC in acute users is shown by time course data that indicate their levels remain high in the critical post-intoxication period when delta 9-THC clears from the blood. Therefore a single-tube assay using solid phase reagents was constructed to quantitate simultaneously delta 9-THC and 9SM. When delta 9-THC values are low or questionable in subject samples, 9SM would appear useful to confirm or refute acute marijuana use. PMID- 6294526 TI - Combined high-performance liquid chromatography and radioimmunoassay method for the analysis of delta 9- tetrahydrocannabinol and its metabolites in plasma and urine. AB - A high-performance liquid chromatography-radioimmunoassay method for the measurement of cannabinoids in plasma and urine is described. The experimental procedure consists of chromatographing a plasma extract or hydrolysed urine sample by high-performance liquid chromatography and quantifying the eluting cross-reacting cannabinoids with radioimmunoassay. The concentrations of delta 9- tetrahydrocannabinol (THC), cannabinol, mono-hydroxylated metabolites, di hydroxylated metabolites, delta 9-THC-11-oic acid, and delta 9-THC-11-oic acid ester glucuronide may be measured by this technique. PMID- 6294527 TI - The use of high-pressure liquid chromatography with electrochemical detection for the assay of nantradol and its application to delta 9- tetrahydrocannabinol. AB - High-pressure liquid chromatography with electrochemical detection permits sensitive, selective and quantitative analysis of biological and pharmaceutical compounds containing electroactive functional groups. In support of pharmacological and clinical studies with nantradol, a potent, nonopioid analgetic, a specific HPLC/electrochemical assay for nantradol and related compounds in plasma was recently developed. In preliminary studies we have now extended this technique to the detection of delta 9- and 11-OH-delta 9- tetrahydrocannabinol. Results are presented which indicate that HPLC coupled to an electrochemical detector provides sensitivity for these compounds in plasma in the low ng/ml range. PMID- 6294528 TI - Quantitative analysis for delta 9-THC, 11-hydroxy-delta 9-THC, and 9-carboxy delta 9-THC in plasma using GC/CI-MS. AB - A method for quantitative measurement of delta 9-THC concentrations in plasma has been developed and applied to the analysis of more than 2000 samples over the past 4 years. The method includes addition of deuterium-labeled delta 9-THC to the plasma for use as the internal standard, a simple and relatively rapid solvent extraction procedure, formation of the trimethylsilyl derivative, and quantitation by selected ion monitoring using ammonia chemical ionization. A similar procedure has also been developed for simultaneous analysis of delta 9 THC and two of its major metabolites in plasma. The latter procedure requires use of a glass capillary column for assays of plasma samples in which the delta 9-THC concentration is below 10 ng/ml. Sensitivities for both methods permit quantitation of THC and its metabolites at concentrations as low as 0.2 ng in 1 ml plasma samples. PMID- 6294529 TI - The roles of individual polyoma virus early proteins in oncogenic transformation. AB - The expression in normal rat cells of modified polyoma virus genomes, separately encoding large T, middle T or small T antigens, has allowed the investigation of the roles of these proteins in oncogenic transformation. Middle T is sufficient to transform cells of established lines but the transformants are serum dependent. Large T lacks intrinsic oncogenic potential but can relieve the serum dependence of normal and transformed cells. Middle T alone cannot transform primary rat embryo fibroblasts. PMID- 6294530 TI - Detection of human papillomavirus DNA in anogenital neoplasias. AB - The presence of papillomaviruses in epithelial-derived cancers from several animal species has led to the speculation that these viruses may also have a pathogenic role in the development of certain human carcinomas, particularly those associated with the anogenital tract. Recently, human papillomavirus (HPV) DNA has been detected in epithelial-derived cancers, both cutaneous and metastatic, from patients exhibiting the rare, chronic flat wart disease, epidermodysplasia verruciformis (EV). Except for patients exhibiting this chronic wart syndrome, the association of HPV genomes with human epithelial cancers has not been demonstrated. In an attempt to delineate the association and possible involvement of papillomaviruses with human anogenital carcinomas, we have begun an analysis of these cancers for the presence of HPV-specific nucleotide sequences by using highly sensitive hybridization procedures capable of detecting distantly related papillomaviruses at low copy number. Here we demonstrate the presence of HPV DNA in several types of anogenital tumours: Bowenoid papulosis, carcinoma in situ, and verrucous carcinoma. These data indicate that HPV can be detected in several types of premalignant and malignant tumours, supporting the contention that this group of viruses may be involved in the development of certain types of human epithelial-derived cancers. PMID- 6294531 TI - Orientation of spin labels attached to cross-bridges in contracting muscle fibres. AB - Electron micrographs showing different cross-bridge orientations in different states of muscle fibres, and X-ray diffraction patterns indicating axial cross bridge disorder in contracting muscle first suggested that force generation in the contracting muscle involved a change in orientation of the myosin heads that form cross-bridges between thick and thin filaments. This has been supported by subsequent work; the myosin molecule has the required flexibility for changes in orientation. The orientation of muscle tryptophans and of probes attached to the myosin heads of permeable muscle fibres depends on the state of the muscle. Recently, fluorescence polarization fluctuations and time-resolved X-ray diffraction patterns have suggested that cross-bridges of a contracting muscle can rotate. We have used electron paramagnetic resonance (EPR) spectroscopy to monitor the orientation of spin labels attached specifically to a reactive sulphydryl on the myosin heads in glycerinated rabbit psoas skeletal muscle. Previously, it has been shown that the paramagnetic probes are highly ordered in rigor muscle, with a nearly random angular distribution in relaxed muscle. We show here that during the generation of isometric tension, approximately 80% of the probes display a random angular distribution as in relaxed muscle while the remaining 20% are highly oriented at the same angle as found in rigor muscle. These findings indicate that a domain of the myosin head does not change orientation during the power stroke of the contractile interaction. PMID- 6294532 TI - [A belly full of bran]. PMID- 6294533 TI - Severe interstitial pneumonitis due to influenza A/Bangkok/1/79. PMID- 6294534 TI - Reactivity of free and coordinated radicals in biology and chemical carcinogenity. I. Coordinated phenoxy radicals generated by hydrogen transfer from hydroxy derivatives of 3,4-benzopyrene. AB - The tautomeric keto form of 6-, 7- and 8-hydroxy-3,4-benzopyrene (BP) prevails in nonpolar solvents at laboratory temperature, in contrast with the enol form of 9 HO-BP and 3-HO-BP, as it was shown according to the ESR study of H-transfer reactions initiated by tert. butyl peroxy radicals coordinated upon the hydroxy derivative of cobalt(III)-acetylacetonate [HO-Co(acac)2]. A further hydroxylation of the keto form of 6-HO-BP, mainly in the position 3, in the thermal interval 40 60 degrees C and the presence of oxygen was observed. Because of lack of steric hindrance in the neighborhood of the position 3 or 9 the primarily formed phenoxy radical after H-abstraction remains stabilized as sigma-coordinated radical on CoIII. Such a radical complex can be destroyed after addition of polar solvents (e.g. methanol). During autooxidation of BP in aerated solutions, similarly as during enzymatic oxidation, a relatively great concentration of high-stable radicals accumulates in nonpolar solvents and in the absence of peroxides. The paramagnetic species is interpreted as a radical pair of two nondissociated semiquinones. In discussion of the carcinogenic activity of oxidative products of BP, not only the enzymatically formed, but also the randomly formed radical intermediates in the first steps of autooxidation must be taken into consideration, which reactivity to biological targets is mediated with the polarity of the biological medium. PMID- 6294535 TI - [Serum concentration of beta-endorphin in schizophrenic and depressive diseases]. PMID- 6294536 TI - A comparison of ranitidine and cimetidine in the treatment of gastric ulcer. A two-centre study. PMID- 6294537 TI - Activities of enzymes involved in purine metabolism and some related adenine nucleotide concentrations of leucocytes in renal failure. AB - We have studied purine metabolism in mononuclear and polymorphonuclear cells from uraemic patients using microradiochemical enzyme assays and high-pressure liquid chromatography. In mononuclear cell lysates the mean activities of adenosine deaminase (EC 3.5.4.4) and 5'-nucleotidase (EC 3.1.3.5) were significantly diminished. The activities of adenylate kinase (EC 2.7.4.3), purine nucleoside phosphorylase (EC 2.4.2.1), adenine phosphoribosyltransferase (EC 2.4.2.7), and hypoxanthine phosphoribosyltransferase (EC 2.4.2.8) were not significantly different in the two groups. The activities of adenosine deaminase and adenine phosphoribosyltransferase were reduced in the polymorphonuclear cell lysates. No clear differences emerged in the concentration of adenine nucleotides in the mononuclear cells. The significance of these changes, which are less marked than those in erythrocytes, is discussed with reference to the immunodeficiency associated with uraemia. PMID- 6294538 TI - Effect of hormones on hepatocyte gluconeogenesis in different models of acute uraemia. AB - Hepatocytes isolated from the livers of starved, sham-operated, bilaterally nephrectomised and ureter-ligated rats as well as rats with ischaemic acute renal failure were used for a comparative study of the effects of different hormones on gluconeogenesis. In all tested groups dibutyryl-3':5'-adenosine monophosphate inhibits glucose synthesis from pyruvate whereas this process is not affected by glucagon and only slightly activated by adrenalin. In contrast, gluconeogenesis from dihydroxyacetone was stimulated by all three hormones at the expense of the conversion of dihydroxyacetone to lactate. In the presence of l-serine adrenalin, glucagon and dibutyryl cAMP also stimulate glucose synthesis, which is more marked in bilaterally nephrectomised and ureter-ligated animals. In half of the experiments with bilaterally nephrectomised rats (group BN 2), lack of sensitivity of hepatocytes to all tested hormones on gluconeogenesis from serine or dihydroxyacetone was observed. The beta-adrenergic antagonist propranolol reduced the stimulatory effect of adrenalin on glucose synthesis from serine and abolished the influence of catecholamines in the presence of dihydroxyacetone and pyruvate. This suggests that both alpha- and beta-receptors are involved in the activation of hepatic gluconeogenesis. Insulin and parathyroid hormone did not change the rate of glucose synthesis in any of the experimental groups. PMID- 6294539 TI - Cyclic nucleotide response to stimulation in isolated glomeruli from dog kidney. AB - The effects of biogenic amines on cyclic 3',5'-adenosine monophosphate (cAMP) and on cyclic 3',5'-guanosine monophosphate (cGMP) were studied in isolated glomeruli from dog kidneys. Of the agents tested, histamine produced the greatest increase in cyclic nucleotide activity. Isoproterenol, dopamine, epinephrine and norepinephrine also produced increases in cAMP and/or cGMP activity but of lesser magnitude. Propranolol decreased cyclic nucleotide levels. Glomeruli isolated from the outer cortex showed greater responses to stimulation than glomeruli from the inner cortex. PMID- 6294540 TI - The role of the neuronal cell body in neurotoxic injury. PMID- 6294541 TI - Conditioned taste aversion following acutely administered acrylamide. AB - Fischer-344 rats were acclimated to a daily 15 min period of water availability. After water consumption had stabilized, the rats were permitted a 15 min access to 0.15% (w/v) solution of saccharin. One to two min following saccharin presentation, they were given various doses of acrylamide (10-50 mg/kg) or lithium chloride (LiCl; 3 mEq/kg) by gavage. Three days later, aversion to the saccharin solution was determined. Acrylamide, like LiCl, produced an aversion to saccharin. The effect of acrylamide appeared to be dose-related with the threshold dose being 10 mg/kg. PMID- 6294542 TI - The subcellular fractionation of the bovine caudate nucleus. AB - Two synaptosomal fractions could be obtained from bovine caudate nucleus on sucrose density gradients one of which had a much greater capacity for 'high affinity' choline uptake than the other but comparable amounts of CAT and choline kinase activity. Specific binding of QNB was widely distributed among all the subcellular fractions except the mitochondrial fraction and in quantitative terms by far the greatest amount was in the microsomal fraction. Only the microsomal fraction contained measurable amounts of glycerophosphocholine phosphodiesterase. PMID- 6294543 TI - Conformational aspects of the actions of some piperidine dicarboxylic acids at excitatory amino acid receptors in the mammalian and amphibian spinal cord. AB - A series of piperidine dicarboxylates (PDA) has been tested for excitatory amino acid agonist and antagonist activity and for synaptic depressant properties of the spinal cords of frogs and immature rats in vitro and of cats in vivo. The substances tested comprised (+/-)-cis-2,3-PDA, (+/-)-cis-2,4-PDA, (+/-)-cis-2,5 PDA, (+/-)-cis-2,6-PDA, (+/-)-trans-2,3-PDA, (+/-)-trans-2,4-PDA and both (+) and (-) forms of cis-2,3-PDA. Peak excitatory amino acid agonist activity was observed with (+/-)-trans-2,3- and (+/-)-trans-2,4-PDA. Excitatory amino acid antagonism and synaptic depressant activity was observed only with cis dicarboxylates, this activity being greatest in the 2,3-analogue. The agonist actions of piperidine dicarboxylates were effectively depressed by the specific NMDA receptor antagonist, (-)-2-amino-5-phosphonovalerate and, where tested, also by D-alpha-aminoadipate and low concentrations of Mg2+. It was concluded that the major part of these agonist actions were mediated by NMDA receptors. The main structural feature of the NMDA agonist actions of these substances was considered to be their close relationship to N-alkyl-aspartic and glutamic acid molecules, with the trans arrangement of the respective 2,3- and 2,4-situated carboxyl groups promoting most effective interaction with the active sites of the NMDA receptor. (+/-)-Cis-2,3-PDA depressed excitatory responses induced by NMDA, kainate, quisqualate, (+/-)-trans-2,3-PDA and (+/-)-trans-2,4-PDA, or evoked by dorsal root stimulation. Both monosynaptic and polysynaptic excitation were susceptible to the depressant action of this substance. The (-) isomer of cis-2,3 PDA carried both excitatory amino acid agonist and antagonist activity and also the synaptic depressant properties observed with the racemic form of this substance. The (+) isomer showed little pharmacological activity. It is proposed that the structure-activity features of these heterocyclic amino acids indicate some of the conformational requirements for interaction with physiological excitatory amino acid receptors. PMID- 6294545 TI - Virus titres and persistently raised white cell counts in cerebrospinal fluid in mice after peripheral infection with demyelinating Semliki Forest virus. PMID- 6294544 TI - IDPN neuropathy in the cat: coexistence of proximal and distal axonal swellings. AB - Administration of beta,beta'-iminodipropionitrile (IDPN) to rodents has previously been shown to produce neurofilament-filled axonal swellings in the proximal regions of motor and sensory nerve fibers. Because of the distinctive distribution of these swellings, IDPN has been classed as a proximal axonopathy and thereby distinguished from other disorders in which similar axonal swellings occur in the distal parts of the axon (distal axonopathies). This report describes the pathology in the peripheral nerves of cats which received intermittent injections of IDPN and calls attention to two previously undescribed pathological changes. First, in addition to the typical proximal swellings associated with IDPN, these animals developed numerous axonal swellings within the distal branches of the sciatic nerve. Distal swellings were present as early as 23 days after initiation of intoxication, indicating that they formed locally (rather than developing in the proximal axon and undergoing transport into the distal regions). The second finding was Wallerian-like degeneration within the affected nerve branches. These changes in the distal sciatic nerve and its branches closely resembled the pathology of the distal axonopathies produced by agents such as the neurotoxic hexacarbons and carbon disulfide. The pathological similarities suggest that IDPN may share with these agents pathogenetic mechanisms to an extent not previously suspected. PMID- 6294546 TI - Electron microscopic study of intranuclear spheres of Bouteille type-III in astrocytes with special reference to liver carcinoma. AB - Astrocytic intranuclear spheres of Bouteille type-III were studied with the electron microscope in the neocortex of 29 patients dying from various causes. The numbers of laminate whorls in these spheres in patients with carcinoma involving the liver were found statistically to be significantly more numerous than the whorls in the spheres of patients who did not have carcinoma of the liver. This second group included patients who were suffering from carcinoma of other organs or from cirrhosis of the liver. The frequency of cases with the spheres increased with age. The numbers of spheres found in people with liver carcinoma, carcinoma of other organs and cirrhosis were the same as the numbers found in people dying from other diseases. The possibility that the increase in numbers of whorls found in people with liver carcinoma was related to a raised metabolic activity of the astrocytes resulting from reduced catabolism of hormones or metabolites by the liver is briefly discussed. PMID- 6294547 TI - The epileptogenic spectrum of opiate agonists. AB - The present authors gave mu, delta, kappa, epsilon and sigma opiate receptor agonists intracerebroventricularly to rats both singly and in combination while monitoring the electroencephalogram from cortical and depth electrodes. Dose response curves were plotted with naloxone against the changes produced by each agonist, and the effect of a number of anticonvulsant drugs on agonist-induced seizures was ascertained. Each opiate agonist produced a different seizure pattern with a different naloxone dose-response curve and anticonvulsant profile. The order of convulsive potency was epsilon greater than delta greater than mu greater than sigma much greater than kappa. Petit mal-like seizure activity was unique to the delta agonist, leucine-enkephalin, while only the mu agonist, morphine produced generalized convulsive seizures. These experiments raise the possibility that opiate systems in the brain may be involved in the pathogenesis of a wide spectrum of seizure disorders. PMID- 6294548 TI - Down-regulation of beta-adrenergic receptors following repeated injections of desmethylimipramine: permissive role of serotonergic axons. AB - The injection of desmethylimipramine (DMI) twice daily for 3 weeks reduced the density of beta-adrenergic receptor recognition sites located in crude synaptic membranes prepared from the cortex and hippocampus and attenuated the stimulation of the membrane-bound adenylate cyclase by isoproterenol. Both actions were abolished if prior to treatment with desmethylimipramine the serotonergic axons were destroyed by an intraventricular injection of 5,7-dihydroxytryptamine. These results show that the down-regulation of beta-adrenergic receptors elicited by repeated injections of desmethylimipramine occurs only if the serotonergic axons are intact. PMID- 6294549 TI - Comparative study of interneuronal relations in the auditory cortex of waking and anesthetized cats. PMID- 6294551 TI - Unit activity of hippocampal structures (the presubiculum). PMID- 6294550 TI - Synaptic responses of sensomotor cortical neurons to stimulation of emotionally significant brain structures. PMID- 6294552 TI - Degree of uniformity of forms of statistical dependence between the EEG and spike trains of neurons. PMID- 6294553 TI - Analysis of power spectra and coherence functions of neocortical potentials during the formation of a postural dominance focus. PMID- 6294554 TI - Program and feedback in organization of the motor system. PMID- 6294555 TI - Structural-functional analysis of identified neurons in the snail Helix pomatia. PMID- 6294556 TI - Integral extracellular postsynaptic potential of a single axon: EEG quantum in the rabbit neocortex. PMID- 6294557 TI - Analysis of cross-correlation functions of cortical unit activity. PMID- 6294558 TI - Histochemical localization of cytochrome oxidase in the hippocampus: correlation with specific neuronal types and afferent pathways. AB - Cytochrome oxidase was histochemically localized in the hippocampus and dentate gyrus of various species of mammals. The most intense staining was observed within stratum moleculare of areas CA1-3 and the outer molecular layer of the dentate gyrus, as well as the somatic and basal dendritic layers of CA3. These regions correspond to the synaptic terminal fields of major excitatory afferent pathways to the hippocampus. The somata of CA3 pyramidal cells and various interneurons were more intensely stained than CA1 pyramidal cells and dentate granule cells, and these levels appeared to correlate positively with their reported rates of spontaneous firing. At the electron-microscopic level, the highest concentrations of densely reactive mitochondria were localized within the distal apical dendritic profiles of principal cells (granule and pyramidal) and certain interneurons (pyramidal basket and stratum pyramidale interneurons). The specific layers in which these structures were found are known to receive intense excitatory input from the perforant pathway. High concentrations of reactive mitochondria were also observed within the somata and proximal dendrites of CA3 pyramidal cells and various interneurons, confirming our light-microscopic observations. These results demonstrated that not only can soma and dendrites of the same cell have disparate but distinct levels of cytochrome oxidase activity, but the pattern of reactivity within a neuron's apical and basal dendrites, or even within specific dendritic segments of the same dendrite can be quite different. While the levels of somatic reactivity correlate with reported levels of spontaneous and/or synaptic activity, the degree of dendritic and somatic staining appeared to be more closely related to the intensity of convergent and/or pathway-specific excitatory synaptic input. PMID- 6294559 TI - Kainic acid receptors and neurotoxicity in adult and immature rat cerebellar slices. AB - The neurotoxic actions of kainate were examined in incubated slices of adult and immature rat cerebellum using light- and electron-microscopy. In the adult, Purkinje cells and inhibitory interneurones became selectively necrotic at concentrations between 5 micro M and 20 micro M. At 30 micro M, granule cells also became affected. In the immature cerebellum, at an age (8 days after birth) when the parallel fibres (thought to use glutamate as transmitter) are largely yet to be developed, selective toxicity was still evident but Purkinje cells and inhibitory interneurones were about 10-fold, and granule cells about 30-fold, less sensitive to kainate than in the adult. Kainate and other excitotoxins also increased cyclic GMP levels in cerebellar slices, apparently through the activation of excitatory amino acid receptors. In the adult tissue, the dose cyclic GMP response curve to kainate was biphasic suggesting the presence of two components. The lower concentrations of kainate eliciting the first component mirrored those inducing selective necrosis of Purkinje cells and inhibitory interneurones while the second component correlated with necrosis of granule cells. Similar correlations applied to the immature cerebellum, but here kainate neurotoxicity appeared to be associated with the activation of receptor types different from those evident in the adult. It is suggested that kainate receptors, whose activation is associated with both neurotoxic damage and elevation of cyclic GMP levels, are located on all cell types in the adult cerebellum, with Purkinje cells and inhibitory interneurones displaying a higher sensitivity to kainate than granule cells. The lower sensitivity of immature cerebellum to the neurotoxic effect of kainate is probably due to lower levels of kainate receptors. PMID- 6294560 TI - Case for diagnosis: adenoid cystic carcinoma of the bronchus. PMID- 6294561 TI - Case for diagnosis: fibrolamellar carcinoma of the liver. PMID- 6294562 TI - [Pneumonectomy for lung cancer. Apropos of 120 cases]. PMID- 6294563 TI - [Conservative surgery of malignant neoplasms of the renal parenchyma and of the upper excretory passages. Prognosis and therapy]. PMID- 6294564 TI - Central projections of the octaval system in the thornback ray Platyrhinoidis triseriata. AB - Octaval nerve centers are studied at medullary and mesencephalic levels after electrical stimulation of the VIIIth nerve and selected brain areas. Recording of evoked potentials shows that primary afferent input is relayed to the anterior, magnocellular, and descending octaval nuclei in the medulla as well as to the cerebellar auricle. The three medullary nuclei project to their contralateral homologues via second-order commissural fibers. Both the ipsi- and contralateral anterior and magnocellular nuclei pass on information via secondary neurons to the oculomotor area and the torus semicircularis in the midbrain. PMID- 6294565 TI - Estimation of generator potentials in primary spindle endings evoked by brief tendon taps in man. AB - The ligamentum patellae in human subjects was tapped by a random triangular waveform, while single primary spindle afferent spikes were recorded from the femoral nerve. Cross-correlation between the primary spindle afferent spikes and the taps revealed a prominent peaked distribution of the discharge probability. The cross-correlograms showed that response time from the onsets of taps to the afferent spike was 7.5 +/- 0.6 msec in 6 units and that the width between the feet of the peaked distribution was 3.3 +/- 0.2 msec in 6 units. The width of the peak implies jitter of the response time of the primary spindle spikes which appear to correspond with the time-to-peak of generator potentials elicited by the taps in primary endings of muscle spindles. Integration of the peaked distribution showed a slow slope at the beginning and near the summit and a steep slope on the way. This curve is assumed to represent the rising phase of the generator potentials in primary spindle endings. PMID- 6294566 TI - Striatal 3-methoxytyramine as an index of dopamine release effects of electrical stimulation. AB - Electrical stimulation of the substantia nigra elicited frequency-dependent increases in striatal 3-methoxytyramine (3-MT). Subsequently, successive increases in dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA) were measured. Inhibition of nigrostriatal cell firing with HA-966 resulted in decreased striatal 3-MT, but in this case elevated DOPAC and HVA were also observed. These data indicate that 3-MT is a reliable index of dopamine (DA) release but that concurrent measurements of DOPAC, HVA and DA provide the most reliable interpretation of drug effects on dopaminergic function. PMID- 6294567 TI - Alpha-melanocyte-stimulating hormone and behavior. AB - Melanocyte-stimulating hormone (MSH) has putative adaptive significance in all forms of species where it is present. In mammals the polypeptide chain influences learning, memory and attention. Chemically MSH shares the first 13 (alpha-MSH) or the first 18 or 22 (beta-MSH) amino acids with adrenocorticotropic hormone (ACTH), even though the mechanisms of secretion and behavioral effects are often quite different. The still shorter peptide chain MSH/ACH4-10 demonstrates significant melanotropic and behavioral actions of alpha-MSH without showing any steroidogenic effect. Behaviorally, alpha-MSH and MSH/ACTH analogues (MSH/ACTH4-9 and MSH/ACTH4-10 influence the parameters of learning, attention and memory in both human and infrahuman subjects. Alpha-MSH has also been reported to increase sensitivity and augment arousal mechanisms in the CNS. Alpha-MSH has been observed to increase and sustain novelty-induced defecation, and this behavior was found to be accompanied by a concomitant decrease in whole brain DA and NE levels in both intact and hypophysectomized rats exposed daily to a test box. The behavioral effects of alpha-MSH may be partially modulated by the enhanced cyclic AMP activity in the CNS observed after MSH administration. MSH also seems to be working in conjunction with the hypothalamic tripeptide MIF-1 and the pineal hormone melatonin, both of which can affect the release of MSH from the pituitary. Recent evidence suggests that MSH is counterbalancing against and complementing with the effects of endorphins, specifically beta-endorphin (61-91 chain of beta-LPH), in maintaining learning and attentive behaviors. PMID- 6294568 TI - Computer assisted analysis of 2-DG autoradiographs. AB - A computerized image processing system is described that assists the neurobiologists in analyzing data from 2-DG autoradiography by providing for: (1) Rapid fine-scale digitization of gray levels using a TV camera (2) The recognition of and verification of subtle differences in optical density with the aid of color windows (3) the superimposition of the autoradiographic image upon the histological image, so that the activity seen in the autoradiograph can be accurately assigned to anatomically defined structures (4) The production of numerical data suitable for statistical analysis and line drawings suitable for black on white publication (5) The relating of local gray level to a norm for the image as a whole, so as to remove the variability introduced by variations in section thickness, in the amount of 2-DG seen by the brain during incorporation, in level of anesthesia, etc. If the localized darkening in autoradiographic images is being used as an index of localized functional activity rather than as a measure of metabolism, normalization obviates the need to obtain arterial blood samples. These routines permit anatomically accurate numerical analysis of autoradiographs without any constraints on the experimental situation. PMID- 6294569 TI - Do neurons process information by relative intervals in spike trains? AB - We suggest the possibility that neurons process information in terms of the relative duration of clusters of adjacent and successive inter-action potential intervals ("bytes" of intervals). If this concept is plausible, as is supported by research from several laboratories which have specifically addressed this possibility, one should be able to see evidence for such patterning in the published illustrations from studies in which this concept was not considered. We present some of this evidence here, along with some illustrations from the original publications. Byte patterns are evident in these examples, even though they went unrecognized by authors and readers alike. It is true that interval patterns are not obvious in all published illustrations of spike trains, and we suggest that this can be explained by one or more of the following: (1) some neurons may operate with an interval-pattern code while others do not, (2) a given neuron may use an interval-pattern code only under certain conditions, and (3) even when such a code exists, it may be difficult to detect for identifiable technical reasons. Therefore, we believe that the relative-internal-pattern concept is a valid scientific hypothesis which merits specific testing of its validity and range of applicability. PMID- 6294571 TI - Nerve terminal effects of indoleamine psychotomimetics on 5-hydroxytryptamine. AB - The mode of action of indoleamine psychotomimetics has been closely linked to 5 HT. Early work showed increases in rat brain levels of 5-HT which were later localized to the nerve-ending fraction. With improved methodology, the 5-HT increment was further detected in the synaptic vesicle fraction. These effects were obtained with several indoleamine hallucinogens but not with mescaline. LSD has been most thoroughly studied and has served as the prototypical compound in ascertaining the mode of action of these drugs. Pretreatment with reserpine abolished the 5-HT effects of LSD in the vesicular fraction. However, a new compartment, termed "juxtavesicular," displayed 5-HT increases following reserpine and LSD. A soluble binding site for 5-HT within the synaptoplasm has been postulated in confirmation of independent results by other groups of investigators. The origin of the 5-HT increment appears to be associated with newly synthesized amine. This was deduced from experiments involving various 5-HT synthesis blockers. To ascertain whether inhibition of raphe neuronal firing is responsible for the accumulation of 5-HT at the nerve terminal, two sets of experiments were performed. Destruction of the raphe cell bodies by radiofrequency lesions failed to abolish the LSD-induced 5-HT increase early after the lesion. Destruction of cortical 5-HT neurons with the neurotoxin 5,7 dihydroxytryptamine completely abolished the 5-HT effect of LSD. It was concluded that an intact nerve terminal is necessary for the expression of the LSD-mediated increases in 5-HT. A LSD "autoreceptor" is postulated, possibly identical to a 5 HT presynaptic receptor inhibiting the release of 5-HT. PMID- 6294570 TI - Opiates, endorphins and the developing organism: a comprehensive bibliography. AB - A comprehensive bibliography of the literature concerned with opiates, endorphins, and the developing organism is presented. A total of 1378 clinical and laboratory references, with citations beginning in 1875, are recorded. A series of indexed accompanies the citations in order to make the literature more accessible. These indexes are divided into clinical and laboratory topics. The clinical section is subdivided into: age of subject examined; maternal aspects; effects on the fetus; pharmacology, physiology, and the withdrawal syndrome; and "other" effects on the offspring. The laboratory section is subdivided into: type of opiate/endorphin studied; species utilized; and major subject areas explored. PMID- 6294572 TI - Estimating unilateral renal function with non-invasive methods. PMID- 6294573 TI - Identification of Warthin's tumor with 99mTc. Report of a case. PMID- 6294574 TI - [Skin grafts in hand surgery]. PMID- 6294575 TI - [Nonchromaffin paraganglioma of the larynx]. PMID- 6294576 TI - Papillary carcinoma of the thyroid occurring as multiple vascular tumors. AB - A case of advanced papillary carcinoma in a 16-year-old boy is presented. While this case is typical of the indolent and locally invasive character of papillary carcinoma, it is remarkable for the extent of tumor involvement appearing as reactive lymphadenopathy. The pronounced tumor vascularity and the similarity of the needle biopsy specimen to chemodectoma both conflicted with the initial diagnostic impression. The correct diagnosis was confirmed only with definitive surgical therapy. PMID- 6294577 TI - Small cell carcinoma of the nose and paranasal sinuses. PMID- 6294578 TI - [Effect of inhibitors of proteolysis on the concentration of cationic proteins and myeloperoxidase in peripheral blood neutrophils in experimental peritonitis in the mouse]. PMID- 6294579 TI - [Role of heparin in increasing the nonspecific resistance of the body]. PMID- 6294580 TI - Paravertebral widening in a case of recurrent Wilms' tumour. AB - In infants and children the association of paravertebral widening and abdominal tumour has been regarded as typical of neuroblastoma. However, a paraspinal mass may develop in other malignant tumours too, but seems to be extremely rare in nephroblastoma. The fourth case so far on record is reported. Microscopically the tumour was of the sarcomatous clear cell variant. PMID- 6294581 TI - Effect of parathyroid hormone on cAMP and 1,25-dihydroxyvitamin D formation and renal handling of phosphate in vitamin D-dependent rickets. AB - Studies were carried out to compare the effects of parathyroid extract (PTE) on the serum concentration of 1,25-dihydroxyvitamin D (1,25[OH]2D), 24,25 dihydroxyvitamin D (24,25[OH]2D), 25,26-dihydroxy vitamin D (25,26[OH]2D) and cAMP, and the urinary excretion of calcium, phosphorus, and cAMP in two normal adult subjects, and in a girl with vitamin D-dependent rickets. The concentration of 1,25[OH]2D was markedly decreased even when she was receiving a daily dose of 25,000 IU of ergocalciferol. PTE infusion resulted in a prompt and distinct increase in the serum levels and the urinary excretion of cAMP in the patient and control subjects. In the control subjects the serum concentration of 1,25[OH]2D increased after the PTE infusion, whereas there was no response in the patient with vitamin D-dependent rickets. The two other dihydroxylated metabolites of vitamin D showed no consistent response to the PTE infusion in the control subjects or the patient. The patient showed no phosphaturic response to PTE while she was receiving high-dosage ergocalciferol treatment. By contrast, when the patient was re-studied after therapy with 1 alpha-hydroxyvitamin D, PTE infusion resulted in an increase in urinary phosphate excretion. These findings might lend support for the notion that 1,25[OH]2D has an effect on tubular phosphate resorption and has a permissive role in the phosphaturic effect of parathyroid hormone. The present findings also confirm that the formation of 1,25[OH]2D is impaired in vitamin D-dependent rickets and indicate that the renal 25 hydroxyvitamin D-1 alpha-hydroxylase is unresponsive to the stimulatory effect of parathyroid hormone in this condition. PMID- 6294582 TI - [Cyclic AMP content in children with gastroduodenitis]. PMID- 6294583 TI - [Effect of nitrogen-siliceous thermal waters on the autoimmune reactions in children with chronic tonsillitis treated at the "Kul'dur" health resort]. PMID- 6294584 TI - [Neonatal cecal perforation. Clostridium perfringens enterocolitis with septicemia and hemolysis. Total megacolon]. PMID- 6294585 TI - Isolated bovine ventricular myocytes. Characterization of the action potential. PMID- 6294586 TI - Calcium currents of isolated bovine ventricular myocytes are fast and of large amplitude. PMID- 6294587 TI - Batrachotoxin protects sodium channels from the blocking action of oenanthotoxin. AB - 1. The blocking action of oenanthotoxin (OETX) and butanol on Na+ channels was studied in voltage clamp experiments on single myelinated nerve fibres treated by batrachotoxin (BTX). 2. OETX (40 microM) blocked Na+ currents through normal channels but did not affect significantly the BTX modified Na+ current. 3. BTX removed the depolarization-induced charge immobilization and slowed down significantly the OFF charge movement. However, the maximum charge displaced, as well as the kinetics of the ON charge movement during a strong membrane depolarization, remained unchanged. 4. OETX blocked the charge movement in normal Na+ channels but did not affect noticeably the charge movement modified by BTX. 5. BTX did not modify the K+ currents and did not protect them from the blocking action of OETX. 6. Butanol (0.01-0.1 M) decreased almost identically and reversibly both normal and BTX-modified Na+ currents. 7. It is concluded that binding of BTX to its receptor protects the Na+ channel from interaction with OETX but left it accessible to butanol. PMID- 6294588 TI - Magnesium restores high K-induced inactivation of the fast Na channel in guinea pig ventricular muscle. PMID- 6294589 TI - Na and Ca spikes produced by ions passing through Ca channels in mouse ovarian oocytes. AB - Ovarian oocyte membrane of the mouse was found to be excitable. Ca-dependent action potentials, which were blocked by Co2+, indicated the existence of Ca channels. In addition, Na-dependent action potentials were detected in Ca2+-free solution. These Na spikes were insensitive to tetrodotoxin (TTX) and were blocked by Co2+, Cd2+, or La3+, suggesting that the Na+ goes through the Ca channel instead of the Na channel. Such a Na current has not been reported in other eggs. It is concluded that both Na+ and Ca2+ pass through the Ca channels during excitation in mouse ovarian oocytes. PMID- 6294590 TI - Changes in extracellular potassium and calcium in rat cerebellar cortex related to local inhibition of the sodium pump. AB - Extracellular K+, Ca2+, and Na+ ([K+]e, [Ca2+]e, [Na+]e) were recorded with ion selective microelectrodes in the cerebellar cortex of urethane-anesthetized rats. Superfusion of the cerebellum with artificial cerebrospinal fluid containing K strophanthidin (10(-6)-10(-4) mol/l) or other cardioactive steroids, known to be inhibitors of the sodium/potassium pump, had the following effects: elevation of resting [K+]e, reduction of poststimulus K+-undershoots, decrease of resting [Ca2+]e and [Na+]e. For instance, at 3 X 10(-5) mol/l K-strophanthidin within the superfusion solution (the unknown intracerebellar concentration being certainly much smaller), [K+]e was elevated up to 130% and [Ca2+]e reduced to 70% of their resting values. Iontophoretic K+-pulses were enhanced in amplitude at the same time. Control experiments with iontophoretic TMA application demonstrated that the glycoside effects were not due (or in higher concentrations only partly due) to shrinkage of the extracellular fluid volume. When tetrodotoxin (10(-7) mol/l) or Mn2+ (1-3 mmol/l) were additionally superfused, K-strophanthidin effects were qualitatively similar, though quantitatively smaller. This indicates that part of the effects were indirect via neuronal activity evoked by the blockade of the sodium pump. The experiments show that reduction of sodium pump activity in cerebellar cortex has rapid and serious consequences on the distribution of potassium and calcium in the extracellular space, resulting in an alteration of neuronal circuit excitability. PMID- 6294592 TI - Stretch stimulates cyclic nucleotide metabolism in the isolated frog ventricle. AB - An investigation has been made of the effect of stretch on cyclic nucleotide metabolism. The levels of endogenous adenosine 3',5' - cyclic monophosphate (cyclic AMP) and guanosine 3',5' - cyclic monophosphate (cyclic GMP) were measured during different patterns of stretch and release. Stretch to Lmax produced a moderate increase in cyclic AMP and a proportionately greater increase in cyclic GMP; as a result, the ratio cyclic AMP: cyclic GMP fell to around 0.5 x control (unstretched) value. The effects of releasing previously stretched preparations resulted in a decrease in cyclic GMP levels. In contrast, the increase in cyclic AMP level following stretch was not affected by this procedure. Nevertheless, these effects together produced an increase in the ratio cyclic AMP: cyclic GMP, the magnitude of which increased linearly with the amplitude of release. PMID- 6294591 TI - Effect of glucocorticoid treatment on the excitability of rat skeletal muscle. AB - Dexamethasone treatment in the rat produced depolarization of extensor digitorum longus (EDL) muscle fibers but not soleus (SOL) fibers studied in vitro at 23 degrees C. The depolarization of EDL fibers was most prominent after 1 day of treatment (treated -77.5 +/- 1.1 mV, control -87.2 +/- 0.8 mV; mean +/- S.E.), and was associated with elevation of the action potential threshold and reduction of the action potential overshoot. In vivo, or in vitro in chloride-free solution, the resting potential and action potential threshold and overshoot of EDL fibers from glucocorticoid-treated and control rats were similar. Sodium currents were studied with a patch voltage clamp. Glucocorticoid treatment did not alter the voltage dependence of sodium channel activation or inactivation in fast twitch muscle fibers. Maximal inward currents occurred at about -29 mV and half-maximal inward currents at about -50 mV. Sodium channels were half inactivated at about -71 mV. Glucocorticoid treatment did not alter the sarcolemmal resistance or capacitance. We conclude that glucocorticoid treatment does not produce muscle weakness or atrophy by altering the excitability of muscle fibers. PMID- 6294593 TI - [Radioprotective effect of YM-08310 in radiotherapy of cervical cancer]. PMID- 6294594 TI - [Proton radiotherapy facility using a spot scanning method]. PMID- 6294595 TI - The incidence of Aujeszky's disease in Finland. PMID- 6294596 TI - [A case of Adie's syndrome with chronic progressive neuropathy and generalized amyloidosis sparing the nerve]. PMID- 6294597 TI - Polyamine-induced hydrolysis of apurinic sites in DNA and nucleosomes. AB - The ability of different polyamines to catalyze hydrolysis of phosphodiester linkages in apurinic and apyrimidinic (AP) sites has been investigated in supercoiled, relaxed and denatured DNA, and also in core and chromatosome particles. The rate constants for the hydrolysis in the DNAs have been determined. In general the order of effectiveness of the polyamines were: spermine greater than spermidine greater than putrescine greater than cadaverine. A 9 fold difference in rate constants was found between spermine and cadaverine. No difference in the rate of hydrolysis was seen between AP-sites in supercoiled and relaxed DNAs, whereas the rate for the single-stranded DNA and DNA in core and chromatosome particles was only half of that in the double-stranded DNA. All AP-sites in both free DNA and DNA-histone particles were hydrolyzed in the presence of polyamines. For all polyamines, with the exception of spermine, increasing concentration of both Mg++ and salts such as KCl both led to a large decrease in the rate of polyamine-induced hydrolysis of AP-sites. The rate of hydrolysis increased markedly with increasing pH in the pH range pH 6 - pH 11. PMID- 6294598 TI - Gene expression: chemical synthesis of E. coli ribosome binding sites and their use in directing the expression of mammalian proteins in bacteria. AB - Mammalian genes, when inserted into bacterial plasmid or phage DNAs, will not be expressed into the corresponding specific proteins in E. coli unless proper initiations signals required for recognition by E. coli ribosomes are provided. We have studied these signals and chemically synthesized two DNA duplexes each containing different initiation signals. These have been inserted in front of the Simian virus 40 (SV40) small tumor antigen gene (SV40 t gene) at varying distances from the ATG initiation codon prior to its cloning into pBR322 plasmid DNA. Plasmid containing clones carrying either of these two synthetic ribosome binding sites (RBS) at varying distances from the SV40 t gene all produced a 17K protein identical to authentic t antigen by immunologic, electrophoretic and proteolytic digestion analyses. This provides a novel method to ensure the specific expression of any contiguous mammalian gene to be cloned to bacteria, and also a unique in vivo method for studying the structure-function (efficiency) relationship of RBS with specific base changes. PMID- 6294599 TI - Bacteriophage T7 late promoters: construction and in vitro transcription properties of deletion mutants. AB - The construction of plasmids containing T7 class I promoters with deletion mutants was described. Restriction fragments, ending at the Hinf I site located at position -10 in the promoter from 14.8% of the T7 genome, were cloned into pBR322. This produced the deletion of either the left or the right part of the promoter. The in vitro transcription properties of these plasmids were determined. Control plasmids were obtained by cloning wild type class II and class III promoters into pBR322. These plasmids also were used to compare the in vitro transcription properties of the two classes of late promoters. Much of the leftward part of a T7 late promoter can be deleted without abolishing activity, but deletion of the right part eliminates promoter activity. Class II, class III, and the mutated promoters have characteristic responses to changes in ionic strength, exogenous glycerol, and temperature. PMID- 6294600 TI - Synthesis in vitro of full length genomic RNA and assembly of the nucleocapsid of vesicular stomatitis virus in a coupled transcription-translation system. AB - Synthesis of a small amount of 42S RNA in addition to the VSV specific mRNA species was observed in a coupled transcription-translation system containing ribonucleoprotein particles from L cell infected with vesicular stomatitis virus and nuclease-treated ribosomal extract obtained from uninfected HeLa cells. Analysis on a CsCl density gradient showed that the synthesized 42S RNA was associated with newly synthesized by protein as a nucleoprotein of bouyant density of 1.3 g/ml. The 42S RNA and the N protein present in the nucleoprotein were resistant to nuclease and protease, respectively. About 35% of the remaining 65% had a complementary polarity. The evidence presented here demonstrates that both the full length genomic and the complementary RNA are associated with N protein in the in vitro replication process. A template role for the complementary 42S RNA for replication of the genomic RNA is also suggested. PMID- 6294601 TI - Simple, efficient in vitro synthesis of capped RNA useful for direct expression of cloned eukaryotic genes. AB - A simple and efficient method for direct in vitro synthesis of capped transcripts of cloned eukaryotic genes is described. As an example capped transcripts were made from a plasmid containing the human fibroblast interferon gene cloned under the control of a prokaryotic promoter. These transcripts were translated in vivo in Xenopus laevis oocytes and in vitro in reticulocyte and in wheat germ cell free protein synthesizing systems. PMID- 6294602 TI - Nucleotide sequence of a Euglena gracilis chloroplast gene coding for the 16S rRNA: homologies to E. coli and Zea mays chloroplast 16S rRNA. AB - The nucleotide sequence of 16S rDNA from Euglena gracilis chloroplasts has been determined representing the first complete sequence of an algal chloroplast rRNA gene. The structural part of the 16S rRNA gene has 1491 nucleotides according to a comparative analysis of our sequencing results with the published 5'- and 3' terminal "T1-oligonucleotides" from 16S rRNA from E. gracilis. Alignment with 16S rDNA from Zea mays chloroplasts and E. coli reveals 80 to 72% sequence homology, respectively. Two deletions of 9 and 23 nucleotides are found which are identical in size and position with deletions observed in 16S rDNA of maize and tobacco chloroplasts and which seem to be characteristic for all chloroplast rRNA species. We also find insertions and deletions in E. gracilis not seen in 16S rDNA of higher plant chloroplasts. The 16S rRNA sequence of E. gracilis chloroplasts can be folded by base pairing according to the general 16S rRNA secondary structure model. PMID- 6294603 TI - A rat tRNA gene cluster containing the genes for tRNAPro and tRNALys. Analysis of nucleotide sequences of the genes and the surrounding regions. AB - A lambda clone carrying a rat DNA fragment of 11.9 kb was isolated from a rat gene library with total rat tRNA as a probe. Nucleotide sequence analysis revealed that the DNA fragment contained six tRNA genes, three for tRNAPro and three for tRNALys. Of the six genes all but one tRNAPro gene have the same polarity. Each tRNA gene is separated by a DNA region of 0.1 to 3.6 kb. The 5' flanking regions of the six rat genes in the cluster do not have any significant sequence homology, but in the 3'-flanking region, each gene has a short T cluster, which is supposed to be a transcription termination signal. PMID- 6294605 TI - Isolation and characterization of phi X174 mutants carrying lethal missense mutations in gene G. AB - A previously constructed Escherichia coli transformant carrying a functional copy of bacteriophage phi X174 gene G on a plasmid, p phi XG, was used to isolate gene G mutants carrying temperature sensitive and lethal missense mutations. Two of the mutations have been characterized by sequencing: one carries a G --> A transition at residue 2821 producing a Gly --> Ser change in codon 143 of the G spike protein; the other carries an A --> G transition at residue 2678 producing Glu --> Gly change in codon 95. Sequencing DNA from 2 other mutants carrying lethal mutations that are rescued with p phi XG did not reveal any changes in the coding sequence. The lesion is believed to be in the intercistronic region between genes F and G. The adsorption kinetics for these mutants appear to be normal. Their burst size is about 25% that of wild type phi X174 on the host carrying p phi XG. These results along with previous results from the senior author's laboratory demonstrate that p phi XG can be used to rescue any gene G mutant of phi X174 regardless of the nature of the mutation involved. PMID- 6294604 TI - A tandem repeat gene in a picornavirus. AB - Three closely related genes for the small genome-linked protein (VPg) of picornaviruses have been identified by sequence analysis as a tandem repeat in the genome of Foot and Mouth Disease Virus (FMDV), strain O1K. This unusual structure was also found in the genome of strain C1O, belonging to a different FMDV serotype. Predicted biochemical properties of the three VPg gene products are in excellent agreement with the data from protein analysis of a heterogeneous VPg population from a third FMDV serotype, strain A10 (1). Taken together, these data indicate that the VPgs from all three genes function equally well in vivo. This is the first report of a tandem repeat gene in a viral genome. PMID- 6294606 TI - Directed mutagenesis of DNA cloned in filamentous phage: influence of hemimethylated GATC sites on marker recovery from restriction fragments. AB - Gapped duplex DNA molecules of recombinant genomes of filamentous phage are constructed in vitro. Denatured restriction fragments covering (part of) the precisely constructed gap are hybridized to the gapped duplex DNA molecules to form ternary duplices. The two strands of the ternary duplex molecules carry different genetic markers within the region spanned by the restriction fragment leading to a one base pair mismatch or to an insertion loop of 93 nucleotides, respectively. The two strands also vary with respect to A-methylation in GATC sites. In cases of asymmetrical methylation, transfection of E. coli with these heteroduplex molecules leads to marker recoveries with a pronounced bias in favour of the marker encoded by the methylated strand. This effect at least partly explains the comparably low marker yields achieved in previous directed mutagenesis experiments using filamentous phage as the vector. The results suggest how these procedures can be optimized. Precise construction of a 93 bp insertion of 9.5% marker yield is described. PMID- 6294607 TI - A new restriction endonuclease from Citrobacter freundii. AB - CfrI, a new restriction endonuclease of unique substrate specificity, has been isolated from a Citrobacterfreundii strain. The enzyme recognizes a degenerated sequence PyGGCCPu in double-strand DNA and cleaves it between Py and G residues to yield 5' -protruding tetranucleotide ends GGCC. PMID- 6294609 TI - The ILtat 1.4 surface antigen gene family of Trypanosoma brucei. AB - The cDNA sequence for the variable surface glycoprotein (VSG) expressed in Trypanosoma brucei clone ILtat 1.4 (called clone D for brevity) hybridizes strongly to three regions in trypanosome genomic DNA. These three regions were extensively characterized by Southern hybridization analyses, genomic DNA cloning and DNA sequence determinations. All three regions occur in the genomes of all trypanosome clones of the ILTAR 1 repertoire regardless of whether or not VSG D was being expressed. Extensive (clone dependent) DNA rearrangements and a (clone independent) double strand DNA break were found distal to the 3'-end of the VSG D coding sequence of one of the regions. VSG D mRNA is most likely synthesized from this region, but a recombinant DNA clone of the VSG coding sequence could not be obtained for confirmation. Recombinant clones of the other two regions were obtained. DNA sequence analyses revealed that their coding sequences differ from each other by 17%. They differ from the ILtat 1.4 cDNA sequence by 4% in one case, and 13% in the other. By analogy with another VSG gene system, one of these two regions may have originally given rise to the third region from which the mRNA is probably transcribed. PMID- 6294608 TI - E. coli initiator tRNA analogs with different nucleotides in the discriminator base position. AB - The effect of base changes at the fourth position from the 3'-terminus of Escherichia coli initiator tRNAMet has been studied to test the 'discriminator hypothesis' which proposed that the nucleotide in this position might have a role in the specificity of the aminoacylation reaction. E. coli initiator tRNA lacking the 3'-terminal tetranucleotide was prepared by partial digestion with S1 nuclease. To construct tRNA analogs with different bases in the fourth position this truncated tRNA was joined by RNA ligase to each of four chemically synthesized 2',3'-ethoxy-methylidene tetranucleotides pACCA(em), pCCCA(em), pGCCA(em), and pUCCA(em). In vitro aminoacylation studies showed that all four molecules accepted methionine, albeit with different Vmax values. PMID- 6294610 TI - Gene amplification in methotrexate-resistant mouse cells. IV. Different DNA sequences are amplified in different resistant lines. AB - DNA was purified from double minutes isolated from MTX-resistant EL4/8 mouse lymphoma cells, digested to completion with Bam H1 restriction endonuclease and cloned in lambda-1059. The properties of the library suggest that the DNA from which it was made was not detectably contaminated with non-dm chromosome material, and that the library is essentially complete for sequences contained in Bam H1 restriction fragments between 9 and 19 kb. The inserts of some selected lambda-recombinants were subcloned in pBR328 or pAT153 to separate sequences of differing repetition frequency. Clones representative of different classes of sequences were used as probes to Southern transfers of Bam H1 digested total nuclear DNAs of various MTX-resistant cell lines. The results clearly show that the amplified unit of each cell line has a unique structure, and that different amplified units differ widely in their sequence composition. PMID- 6294611 TI - Drosophila mitochondrial DNA: a novel gene order. AB - Part of the replication origin-containing A+T-rich region of the Drosophila yakuba mtDNA molecule and segments on either side of this region have been sequenced, and the genes within them identified. The data confirm that the small and large rRNA genes lie in tandem adjacent to that side of the A+T-rich region which is replicated first, and establish that a tRNAval gene lies between the two rRNA genes and that URF1 follows the large rRNA gene. The data further establish that the genes for tRNAile, tRNAgln, tRNAf-met and URF2 lie in the order given, on the opposite side of the A+T-rich region to the rRNA genes and, except for tRNAgln, are contained in the opposite strand to the rRNA, tRNAval and URF1 genes. This is in contrast to mammalian mtDNAs where all of these genes are located on the side of the replication origin which is replicated last, within the order tRNAphe, small (12S) rRNA, tRNAval, large (16S) rRNA, tRNAleu, URF1, tRNAile, tRNAgln, tRNAf-met and URF2, and, except tRNAgln, are all contained in the same (H) strand. In D. yakuba URF1 and URF2, the triplet AGA appears to specify an amino acid, which is again different from the situation found in mammalian mtDNAs, where AGA is used only as a rare termination codon. PMID- 6294613 TI - Ribosomal RNA genes of Trypanosoma brucei. Cloning of a rRNA gene containing a mobile element. AB - An ordered restriction map of the ribosomal RNA genes of Trypanosoma brucei brucei is presented. Bgl II fragments of T.b.brucei genomic DNA were cloned into pAT 153, and the clones containing rDNA identified. Restriction maps were established and the sense strands identified. One clone was shown by heteroduplex mapping to contain a 1.1 kb inserted sequence which was demonstrated to be widely distributed throughout the genomes of members of the subgenus Trypanozoon. However, in two other subgenera of Trypanosoma, Nannomonas and Schizotrypanum, the sequence is far less abundant. Analysis of the genomic DNA from two serodemes of T.b.brucei showed that the sequence was present in the rRNA of only one of them, implying that the sequence is a mobile element and that its appearance in rDNA is a comparitively recent occurrence. PMID- 6294612 TI - The construction of cosmid libraries which can be used to transform eukaryotic cells. AB - Cosmid vectors have been developed which carry selective markers for growth in bacteria (beta lactamase gene) and animal cells (the Herpes Simplex virus thymidine kinase gene, the transposon Tn-5 aminoglycosyl 3' phosphotransferase gene and the E. coli guanine phosphoribosyltransferase gene). The design of the cosmids allows the exchange of the eukaryotic markers in recombinant cosmids. Human and mouse cosmid libraries containing DNA inserts of about 40kb have been generated by an improved method. Several clones from the human beta-globin locus were isolated. These cosmids transform mouse L cells at high efficiency in both circular and linear form. The newly introduced genes are expressed accurately in L cells. PMID- 6294615 TI - Identification of splicing signals in introns of yeast mitochondrial split genes: mutational alterations in intron bI1 and secondary structures in related introns. AB - Four mitochondrial mutations are known to block excision of intron I1 of the cob gene in S.cerevisiae. The nucleotide sequence alteration of one of them, M4873, has been determined. It is a deletion of 1 bp in a run of five G's at a distance of 30 to 34 bp upstream to the 3' splice point. Reversion is found to occur by restoration of the run of five G's either by insertion of 1 G (wild type reversion) or by transition A leads to G next to this run of G's (pseudo-wild type reversion). The effect of mutation and reversion on RNA splicing indicates that the run of five G's is of critical importance for intron I1 excision, possibly in participating in the formation of a splice signal with a helical structure. This presumption is confirmed by the observation that this sequence is part of a larger sequence of some 80 bp next to the 3' splice point which is conserved to some extend in the four mitochondrial introns (bI1, aI1, aI2, aI5) that survive after excision as circular RNAs. Most striking is the conservation of this sequence at the level of secondary structure. PMID- 6294614 TI - Hypoxanthine-guanine phosphoribosyltransferase genes of mouse and Chinese hamster: construction and sequence analysis of cDNA recombinants. AB - Recombinant plasmids containing DNA inserts complementary to mRNA coding for hypoxanthine-guanine phosphoribosyltransferase (HPRT) from mouse and Chinese hamster cell lines have been isolated from cDNA libraries and characterized by DNA sequence analysis. A total of 1292 nucleotides of the mouse cDNA sequence and 1301 nucleotides of the Chinese hamster cDNA sequence has been determined. Each of these sequences includes an open reading frame of 654 nucleotides (218 amino acids) corresponding to the HPRT protein coding region. The deduced amino acid sequences for the mouse and Chinese hamster enzymes are presented and compared to that of human HPRT. At least 95% of the amino acids are conserved in the three species. In addition, we present evidence that two species of HPRT mRNA, which differ in the site of polyadenylation that is utilized during processing of the RNA transcripts, exist in Chinese hamster cells. PMID- 6294616 TI - The binding of gyrase to DNA: analysis by retention by nitrocellulose filters. AB - Three distinct Escherichia coli DNA gyrase complexes with DNA can be identified using a nitrocellulose filter-binding assay. One complex consists of an ensemble of two subunit A and two subunit B protomers bound noncovalently to specific sequences of DNA. High levels of each subunit alone are inactive but a single gyrase molecule binds DNA to a filter. At 23 degrees, the complex has a dissociation constant of approximately 10(-10) M and a half-time of decay of about 60 h. It is sufficiently stable that it can be purified by gel filtration and retain full supercoiling activity. Gyrase binds preferentially to relaxed DNA over supercoiled DNA by a factor of about 10. On addition of oxolinic acid, a second complex is formed that is distinguished by its stability in high ionic strength solutions and by efficient conversion to a third form upon addition of protein denaturants. The first and second complexes require Mg++ for optimal formation. The third form has been shown previously to contain denatured A protomers covalently linked to DNA that is broken at the site of attachment. PMID- 6294617 TI - Initiation and termination of the bacteriophage phi X174 rolling circle DNA replication in vivo: packaging of plasmid single-stranded DNA into bacteriophage phi X174 coats. AB - The bacteriophage phi X174 viral (+) origin when inserted in a plasmid can interact in vivo with the A protein produced by infecting phi X174 phages. A consequence of this interaction is packaging of single-stranded plasmid DNA into preformed phage coats resulting in infective particles (1). This property was used to study morphogenesis and to analyse the signals for initiation and termination of the rolling circle DNA replication in vivo. It is shown that the size of the DNA had a strong effect on the encapsidation by the phage coats and the infectivity of the particle. Termination was analysed by using plasmids with two phi X (+) origins either in the same orientation or in opposite orientation. Both origins were used with equal frequency. Initiation at one origin resulted in very efficient termination (greater than 96%) at the second origin in the case of two origins in the same orientation. When the two (+) origins have opposite orientations, no correct termination was observed. The second origin in the opposite strand effectively inhibits (greater than 98%) the normal DNA synthesis; i.e. the covalently bound A protein present in the replication fork interacts with the (+) origin sequence in the opposite strand. PMID- 6294619 TI - The temporal order of replication of murine immunoglobulin heavy chain constant region sequences corresponds to their linear order in the genome. AB - The time of replication during the S phase in a murine erythroleukemia (MEL) cell line was determined for immunoglobulin heavy chain constant region C alpha, C gamma 2b and C mu sequences whose boundaries are defined by EcoR1 restriction endonuclease sites (EcoR1 segments). Logarithmically growing cultures of MEL cells with an S phase of about 7.5 hours were pulse labelled with 20 micrograms/ml of 5-bromodeoxyuridine (BUdR). The cells were then fractionated by centrifugal elutriation into 10-12 distinct populations containing cells in different stages of the cell cycle. Flow microfluorimetric (FMF) analysis of DNA content, measurements of cell volume and autoradiography after 3H-thymidine pulse labelling were used to determine position in the cell cycle. Fractions were pooled to represent four selected intervals of S in which BU-DNA was synthesized for 2.5 hrs or less. Newly replicated DNA which had incorporated BUdR into one strand was isolated, cleaved with EcoR1, and separated on neutral Cs2S04 gradients. Equal amounts of BU-DNA replicated during these four intervals of S were electrophoresed in 0.8% agarose gels, transferred to diazotized aminobenzyloxymethyl paper and hybridized with 32p probes containing the C alpha, C gamma 2b and C mu genes and flanking sequences. The relative amounts of segments replicated were assessed by quantitation of the appropriate bands on the autoradiograms by microdensitometry. The results indicate that the 2.8 kb C alpha, 6.6 kb C gamma 2b and 12 kb C mu EcoR1 segments in these MEL cells replicated during defined intervals of the first half of the S phase. The order of replication of these EcoR1 segments as the cells proceeded through S was C alpha, C gamma 2b, C mu, corresponding to the linear order of the genes determined by restriction endonuclease mapping. PMID- 6294618 TI - Structure and transcription of the spinach chloroplast rDNA leader region. AB - A cloned fragment of spinach chloroplast DNA carrying 140 bp of the 16S rRNA gene and 691 bp upstream this gene has been analysed by DNA sequencing, by in vitro transcription, by S1 mapping with chloroplast RNAs and purified 16S rRNA from 30S ribosomal subunits. A tRNAVal gene has been located between the position 394 and 465. Crude chloroplast RNA polymerase has been purified by heparin sepharose chromatography of a 80 000 g supernatant from pure lysed spinach plastids and used to transcribe the cloned Bg1 II-Pvu II DNA fragment. Four in vitro transcripts of about 830, 550, 350 and 260 bases were obtained whatever RNA polymerase used: the chloroplast or the E. coli enzyme. The transcripts of 550 and 260 bases are initiated by ATP. S1 mapping with in vivo chloroplasts RNAs on 5' labelled separated strands from Bg1 II-Pvu II fragments indicates 2 protected DNA fragments respectively of 140 and 260 bases on the strand which codes for rRNAs and possibly one protected DNA fragment of 550 bases on the other strand. The start site of the 260 bases transcript might correspond to the initiation site of transcription of the rRNA genes. The possibility that the 550 bases transcription of the non coding strand for rRNA genes corresponds to the beginning of a mRNA is discussed. PMID- 6294620 TI - Sequence signal involved in the generation of an internally deleted defective interfering RNA from vesicular stomatitis virus. AB - We have determined the nucleotide sequence at the 3' end of the vesicular stomatitis virus (VSV) polymerase or L gene and compared it to that obtained from a defective interfering particle (DI) RNA generated by this virus. The latter (DI 0.50) contains a large internal deletion within this gene. The deletion begins exactly 253 residues from the transcription start of the gene and extends to approximately 300 bases from the 5' end of the standard RNA genome. The flanking sequences bear no homology to the eukaryotic consensus splice sequence. The sequence immediately preceding the deletion is complementary to the ribosome binding site of the L gene transcript and also resembles genome transcription termination sites. In addition, we present the results of nuclease protection experiments which show that this DI RNA retains an exact copy of the 3' end half of the standard genome (leader, N, NS, M and G genes), although its own 3' end is non-genomic. The implications of these findings regarding mechanisms of DI generation are discussed. PMID- 6294621 TI - Nucleotide sequence of the 3' half of AKV. AB - We report the nucleotide sequence of the 3' half of the ecotropic murine leukemia virus AKV genome. To obtain a preliminary sequence, we developed a sequencing strategy whereby a nested set of restriction fragments is chemically modified prior to gel purification and strand scission. The sequence defines the genetic map of the 3' half of AKV and locates recombinant regions previously identified in structural analyses of MCF viruses. PMID- 6294622 TI - Drosophila topoisomerase I: isolation, purification and characterization. AB - We have purified and characterized topoisomerase I from Drosophila melanogaster. The molecular weight of the enzyme is 135,000; 100,000, 90,000, and 65,000 molecular weight products result from degradation of the enzyme. The enzyme relaxes both positive and negative supercoiled DNA. Mg++ is not absolutely required, but stimulates the enzymatic activity considerably. PMID- 6294623 TI - Primary structure of the ompF gene that codes for a major outer membrane protein of Escherichia coli K-12. AB - The nucleotide sequence of the ompF gene coding for a major outer membrane protein of Escherichia coli K-12 has been determined and the amino acid sequence of the OmpF protein was deduced from it. The OmpF protein contains 340 amino acid residues, and is produced from a precursor having 22 extra amino acid residues, the signal peptide, at the amino terminus. The expected secondary structure of the OmpF protein had a high beta-sheet content with a low alpha-helix content. The promoter region and the transcription termination region of the ompF gene had a significantly high AT content, while the AT content of the coding region was about the same as the average AT content of the E. coli chromosome. Following the termination codon, a typical rho-independent transcription termination signal was observed. The codon usage in the ompF gene was highly nonrandom; the codons preferably utilized are those recognized by the most abundant species of isoaccepting tRNAs or those, among synonymous codons recognized by the same tRNA, that can interact more properly with the anticodon. PMID- 6294626 TI - [Prognostic value of the assessment of nonspecific immunologic reactivity in lung cancer]. PMID- 6294625 TI - Multiple Pol I initiation sequences in rDNA spacers of Drosophila melanogaster. AB - Sequence analysis of the rDNA nontranscribed spacer (NTS) of D.melanogaster shows that it contains many regions that are perfectly homologous with the Pol I transcription initiation site. Each initiation site homology is contained within a 240 base pair repeated sequence which are organised in a tandem array terminating 140 bases pairs upstream from the transcription initiation site. We discuss the significance of these regions and the maintenance of multiple homologies against the observed high rate of spacer divergence between related Drosophila species. In addition there are two other regions of repetition in the NTS: a 90 base pair periodicity commencing 16 bases downstream from the 3' end of the 28S gene and a number of 340 base pair repeats. A similar complex of different periodicities has been described in Xenopus and may be a general feature of NTS sequences. PMID- 6294624 TI - Neuraminidase gene from the early Asian strain of human influenza virus, A/RI/5 /57 (H2N2). AB - The complete structure of the neuraminidase gene from the A/RI/5-/57 strain of influenza virus has been determined. It is 1467 nucleotides long and codes for a protein of 469 amino acid residues. Comparison with the gene sequence for the N1 strains A/WSN/33 and A/PR/8/34, the N2 strain A/Udorn/72 and the protein sequence for the N2 strain A/Tokyo/3/67 shows the amino acid sequence changes that have occurred during antigenic shift (60%) and drift (7-9%). PMID- 6294627 TI - New agents in diarrhea. PMID- 6294628 TI - The third generation cephalosporins and the pediatric practitioner. PMID- 6294630 TI - Eating, drinking and temperature responses to intracerebroventricular cholecystokinin in the chick. AB - The central effect of cholecystokinin-octapeptide (CCK), SQ 19,844 or sincalide, on the intake of food and water and on colonic temperature (Tc) was investigated using the broiler cockerel. Four-week old chicks were maintained in a thermoneutral environment of 23-24 degrees C. After food was removed for a 24 hr interval, CCK was infused in a volume of 10.0 microliters into the lateral cerebral ventricle (ICV) in doses ranging from 10-150 ng. Although lower doses of CCK had no effect on food intake, 100 or 150 ng of CCK significantly reduced consumption of food in a dose-dependent manner; water drinking was significantly decreased by 100 ng of CCK. In addition, CCK at doses of 100 and 150 ng prevented the slow rise in Tc observed following infusions of control CSF. This latter effect appeared to be a result of feeding activity associated with caloric intake and the heat increment in the control birds rather than a specific thermoregulatory effect. Overall, our results suggest that CCK may comprise a part of the central mechanism underlying the neural control of short term satiety in an avian species similar to that proposed for the mammal. PMID- 6294629 TI - Effects of MSH/ACTH 4-10 on the classically-conditioned rabbit nictitating membrane. AB - Subjects were conditioned/extinguished under four experimental conditions using either MSH/ACTH 4-10(A) or diluent (D): D/D, D/A, A/D, and A/A. The major question investigated was whether or not the peptide has an effect on this classically-conditioned behavior similar to that reported for instrumental conditioning paradigms. The results indicated that it does not. An effect was seen on performance, not on learning or attentional processes. Animals treated with the peptide performed more poorly (i.e., displayed fewer conditioned responses) during both acquisition and extinction. In addition, there was an apparent residual effect of the peptide that lasted 24 but not 48 hours. PMID- 6294631 TI - Autoradiographic distribution of [3H]neurotensin receptors in rat brain: visualization by tritium-sensitive film. AB - [3H]Neurotensin ([3H]NT) binds specifically to a single class of binding sites on slides-mounted sections of rat brain 1Kp = 5.1 nM; Bmax = 16.2 fmol/mg tissue). Bound [3H]NT can be displaced by nonradioactive NT and a series of its fragments and analogues with relative potencies that correlate closely (r = 0.89; p less than 0.01) to their potencies in the rat stomach strip bioassay. These results suggest that NT receptors are similar in both systems. [3H]NT binding sites were visualized by using tritium-sensitive LKB film analysed by computerized densitometry. [3H]NT receptors are highly concentrated in the external layer of the olfactory bulb, in the rhinal sulcus, in certain nuclei of the amygdala, in the substantia nigra, zona compacta and in the ventral tegmental area. The high density of [3H]NT receptors in the last two areas suggest an interaction between NT and brain dopaminergic systems such as the nigrostriatal and the mesolimbic pathways. PMID- 6294632 TI - Localization of neurotensin binding sites in rat kidney. AB - [3H]Neurotensin ([3H]NT) appears to bind specifically to a single class of sites in slide-mounted rat kidney sections (KD = 8.3 nM; Bmax = 31.6 fmol/mg tissue). Bound [3H]NT can be displaced by nonradioactive NT and a series of its fragments and analogues with relative potencies that correlate well (r = 0.91; p less than 0.005) to their potencies in the rat stomach strip bioassay. These results suggest that NT receptors are similar in both systems. However, they are probably slightly different from those present in the guinea pig atria (r = 0.78; p less than 0.1). We visualized these sites by using the tritium-sensitive LKB film technique analysed by computerized densitometry. [3H]NT binding sites are highly concentrated in the renal cortex while low levels are observed in the renal medulla. The possible physiological and/or pathophysiological significances of the presence of [3H]NT binding sites in the kidney are discussed. PMID- 6294633 TI - Immunohistochemical and immunocytochemical localization of gamma-melanocyte stimulating hormone (gamma-MSH)-like immunoreactivity in human and rat hypothalamus. AB - Immunohistochemical localization of gamma-MSH was studied in human and rat hypothalamus by peroxidase-labeled antibody method both at light and electron microscopic levels. Human and rat hypothalamus contained immunoreactive gamma-MSH neurons and varicose nerve fibers. The distribution of gamma-MSH-positive nerve fibers was similar to that of beta-endorphin previously reported. By our "re staining method," gamma-MSH and ACTH were localized in the same neurons and nerve fibers. In the rat, the immunologic staining of gamma-MSH in hypothalamic neurons and nerve fibers was not diminished after hypophysectomy. These findings strongly suggest the possibility of actual precursor production in the hypothalamus which is similar to that in the anterior pituitary. The presence of gamma-MSH at the synapse-like structure of the nerve terminal may indicate that gamma-MSH could function as a neurotransmitter or a neuromodulator. PMID- 6294634 TI - Characterization of Leydig cell gonadotropin-releasing hormone binding sites utilizing radiolabeled agonist and antagonist. AB - Gonadotropin-releasing hormone (GnRH) binding sites in intact Leydig cells and in membrane preparations were investigated using 125I-labeled GnRH agonist and antagonist. Binding was saturable and involved a single class of high affinity sites. Intact Leydig cells and a membrane preparation had a higher affinity for GnRH agonist (Kd 3.0 +/- 1.7 X 10(-10) M) than for GnRH antagonist (Kd 10.0 +/- 1.8 X 10(-10) M). With anterior pituitary membranes the Kd was 2.8 +/- 0.7 X 10( 10) M for the agonist and 2.4 +/- 1.4 X 10(-10) M for the antagonist. The Kd for GnRH was similar for Leydig cells and the anterior pituitary. Chymotrypsin and trypsin digestion decreased receptor binding, but neuraminidase increased Leydig cell binding in contrast to the decrease in binding observed with pituitary receptors. The results suggest that the Leydig cell GnRH binding sites may differ from the pituitary receptor which may be related to structural differences in GnRH-like peptides recently described in extracts of rat testis. PMID- 6294636 TI - Lack of effect of corticotropin releasing factor on hypothalamic dopamine and serotonin synthesis turnover rates in rats. AB - Catecholamine and serotonin neurons in the hypothalamus regulate the secretion of corticotropin releasing factor (CRF). We considered the possibility that CRF might in turn affect the activity of these aminergic neurons. We examined the effect of intracisternal administration of synthetic CRF on the synthesis turnover rates of dopamine and serotonin in the hypothalamus of adult male rats using two different methods to assess turnover. In one study, we measured the accumulation of L-dihydroxyphenylalanine (L-DOPA) or 5-hydroxytryptophan (5-HTP) in mediobasal hypothalamus after L-aromatic amino acid decarboxylase inhibition with m-hydroxybenzylhydrazine 20 min before sacrifice, and in the second study we measured the accumulation of dopamine, norepinephrine, epinephrine and serotonin after monoamine oxidase inhibition with pargyline 20 min before sacrifice. The commercial CRF which we administered intraarterially increased plasma ACTH and corticosterone concentrations. Intracerebral CRF 5 to 20 micrograms 20 min before sacrifice or 20 micrograms 110 min before sacrifice did not alter the m hydroxybenzylhydrazine-induced accumulation of L-DOPA or 5-HTP when compared with saline vehicle-injected controls. CRF 20 micrograms did not alter basal concentration or pargyline-induced accumulation of the catecholamines or serotonin in whole hypothalamus when compared with saline vehicle-injected controls. Thus, intracisternal administration of CRF did not alter hypothalamic dopamine or serotonin synthesis rates as assessed by two nonsteady state turnover methods. The data suggest that the release of CRF from neurons in hypothalamus does not alter the activity of catecholamine or serotonin neurons in the hypothalamus of normal adult male rats. PMID- 6294635 TI - Antinociceptive potencies of beta-casomorphin analogs as compared to their affinities towards mu and delta opiate receptor sites in brain and periphery. AB - beta-Casomorphins and their analogs were tested for their opioid activities in the myenteric plexus longitudinal muscle preparation of the guinea pig ileum (GPI), the isolated mouse vas deferens (MVD), and for their affinities to mu- delta- and kappa- binding sites in rat brain membranes. C-terminal amidation of beta-casomorphin-4 and (-5) increased opioid potency in both organ preparations (GPI, MVD) and affinity to mu-binding sites in brain whereas binding to delta sites was diminished. These beta-casomorphin-amides displayed a 2-3 times greater naloxone reversible antinociceptive effect than natural beta-casomorphins. Introduction of D-alanine at position 2 in the beta-casomorphin-amides increased potency in the GPI whereas activity in the MVD was only slightly changed. These compounds, however, showed a remarkable increase in binding to delta-sites in brain with an unaffected or slightly increased binding to mu-sites and decreased binding to kappa-sites. D-Ala2-beta-casomorphin-4 and (-5) amides were 10 times more potent antinociceptive agents than corresponding beta-casomorphin-amides. These results suggest firstly, that peripheral delta-receptors in the MVD are not as closely related to delta-binding sites at rat brain membranes as is the case with mu-receptors in the GPI and mu-binding sites, and secondly, in addition to mu-receptors, delta-receptors may be of importance in mediating antinociception. PMID- 6294637 TI - Electrophysiological and contractile responses of canine atrial tissue to adrenocorticotropin. AB - The direct extra-adrenal actions of adrenocorticotropin 1-39 (ACTH) on electrical (E) and mechanical (M) characteristics of canine atrial tissues (AT) were investigated in in vitro experiments. One hundred twenty-five mU/ml of ACTH 1-39 significantly augmented the catecholamine induced positive inotropism as seen by shortening the time to peak tension (10.6%, p = 0.01) and increasing peak isometric tension (3.5 times, p = 0.001). Effects on the M responses were inhibited by propranolol (10(-6) M) (P). ACTH did not significantly modify action potential E or M parameters during cholinergic receptor antagonism or alpha adrenergic receptor antagonism. Existence of a specific ACTH receptor was demonstrated using 125I radioiodinated ACTH 1-24. Significant binding of 125I ACTH to AT was observed. Intracellular C-AMP levels were also measured in AT using radioimmunoassay. Tissues were exposed to 125mU/ml ACTH 1-39 plus combinations of norepinephrine (10(-6) M) (NE) and P. ACTH alone did not elevate intracellular C-AMP levels. NE increased C-AMP levels were not further increased by ACTH. Exposure to antagonist returned elevated C-AMP levels to control values. In conclusion (1) ACTH augments the NE induced M positive inotropism of the beta adrenergic receptor system. (2) ACTH specifically binds to AT and (3) ACTH does not utilize the C-AMP second messenger system. PMID- 6294638 TI - Effects of an LHRH agonist and gonadotropins on testicular prolactin receptors. AB - Effects of administration of the LHRH agonist D-Leu6-LHRH ethylamide (LHRH-A), gonadotropin (PMS), and their interaction on testicular prolactin (PRL) receptor levels were investigated in rats. LHRH-A (2 micrograms/100 g body wt.) or saline was injected SC daily, and PMS (5 IU) injected every other day. In intact rats, the testicular PRL receptor levels were about 400 fmoles/testis after either 1 or 7 daily injections of saline. Administration of LHRH-A decreased PRL receptors to 12% of that of saline-injected control rats at day 1, and to 20% at day 2, and PRL receptor levels were partially restored to 55% at day 7. In hypophysectomized rats given daily injections of saline for 7 days PRL receptor levels were only 20% of those in saline-injected intact rats. Injections of LHRH-A in hypophysectomized animals did not further decrease PRL receptor numbers at this time. Administration of PMS to hypophysectomized rats for 7 days partially reversed the reduction of PRL receptors that occurred after hypophysectomy, to 46% of those in intact controls. Injections of LHRH-A into hypophysectomized. PMS treated animals did not significantly alter PRL receptors on day 1 (117% of that of saline-injected, hypophysectomized, PMS-treated rats at day 1) or day 2 (96% of same-day controls), but decreased PRL receptors on day 7 to 102 fmoles/testis (55% of same-day controls). This latter concentration is nearly the same as that in saline-injected, 7-day hypophysectomized rats not treated with PMS. These findings suggest that: (1) the effects of LHRH-A on testicular PRL receptors differ depending on the presence or absence of gonadotropin, (2) gonadotropin, primarily FSH, maintains some population of testicular PRL receptors, and these gonadotropin-dependent PRL receptors are suppressed by direct action of LHRH-A upon the testes, and (3) there is a population of PRL receptors which is not affected by LHRH-A or gonadotropin. PMID- 6294639 TI - Metkephamid (Tyr-D-ala-Gly-Phe-N(Me)Met-NH2), a potent opioid peptide: receptor binding and analgesic properties. AB - The interaction of metkephamid (Tyr-D-Ala-Gly-Phe-N(Me)Met-NH2) with 3H dihydromorphine and 3H-D-Ala2-D-Leu5-enkephalin binding has been examined in rat brain homogenates. Displacements of both 3H-ligands by metkephamid indicate that metkephamid interacts competitively with greatest potency to the high affinity binding component for both ligands (mu1 site). Unlike most enkephalins and opiates, metkephamid binds equipotently to both morphine-selective (mu2) and enkephalin-selective (delta) binding sites. Metkephamid is differentiated from morphine by its better than 12-fold higher affinity for the delta receptor. Blockade of the high affinity (mu1) binding in vivo with high doses of naloxazone dramatically reduces metkephamid's analgesic potency. PMID- 6294641 TI - [Disorders of arachidonic acid metabolism in experimental arteriosclerosis]. PMID- 6294640 TI - [Effect of drugs modulating the function of adrenergic receptors on gastric emptying in duodenal ulcer]. PMID- 6294643 TI - Nutrition, platelet function, and heart disease. PMID- 6294642 TI - [Clinical evaluation of ifosfamide in one-agent chemotherapy of malignant testicular tumors]. PMID- 6294644 TI - Oral antifungal therapy. Current status of ketoconazole. AB - Ketoconazole, an oral, broad-spectrum antifungal agent with relatively low toxicity, is the preferred drug for treatment of chronic mucocutaneous candidiasis and an alternative choice for treatment of thrush, vaginitis, histoplasmosis, and coccidioidomycosis. Clinical trials have shown it to be effective also against dermatophytoses. Further testing is necessary to determine its effectiveness for treating blastomycosis, cryptococcosis, sporotrichosis, and aspergillosis. PMID- 6294645 TI - [Psychopharmaca in chronic pain]. PMID- 6294646 TI - Fluid shift across the placenta: II. Fetomaternal transfer of horseradish peroxidase in the guinea pig. AB - The fetal side of the isolated guinea-pig chorio-allantoic placenta has been perfused with a fluid containing horseradish peroxidase. Electron micrographs show that protein is transferred across the fetal capillary endothelium via micropinocytosis as well as by transport through intercellular clefts. Under normal hydrostatic pressure no pinocytosis or other signs of fetomaternal protein transfer across the trophoblast have been observed. Instead, macrophages ingest the proteins that have reached the interstitial space between fetal endothelium and trophoblast. When the fetal venous pressure is raised up to 20 mmHg about 30 to 50 per cent of the arterial perfusion fluid, including proteins, is shifted into the maternal lacunae. In the beginning of this process slender tubules, and later on bag-like channels, appear that cross the trophoblast and probably connect lacunae and interstitium. The inner surface of the channels and the maternal lacunar surface are labelled by peroxidase. This indicates a fetomaternal transfer of proteins conveyed by the fluid shift. Protein transfer and fluid shift return to their previous levels when the fetal venous pressure is reduced. The size and number of channels are reduced but their structure and the reversibility of the events refute the possibility that they are of traumatic origin. It cannot be decided whether the channels are pre-existent but unrecognizable, or de-novo formations. PMID- 6294647 TI - [Changes in the cardiovascular system after continuous and intermittent administration of the rodenticide ratron]. AB - The authors follow up the effect of "Ratron" a raticide (coumarin derivative) with two regimes of administration--monotonous and intermittent--upon the state of the cardiovascular system of experimental animals. The experiment was carried out on 65 female albino rats, grouped into three groups: group I--20 animals treated 5 times weekly with 1/100 LD50 ratron--five weeks; group II--25 animals, administered 5 times weekly every second week 1/50 LD50, and group III--20 controls. At the end of the sixth month, the state of the metabolic processes and the qualitative and quantitative characteristic of the connective tissue of myocardium and aorta were followed up by biochemical and histological methods. Unidirectional deviations in metabolism and the structure of myocardium and aorta were observed under the effect of both regimes, the changes being better manifested with the intermittent administration of ratron. Data were established not only about the general toxic effect of ratron but about the direct lesions of vessels, multiple hemorrhages, interstitial oedema, regeneration of endothelium, structural changes in the small vessels, changes in the components of the basic intercellular substance of the connective tissue. PMID- 6294648 TI - [Histochemical mechanisms in etiology of pulmonary fibrosis after short-term exposure to a mixture of dust and quartz from copper mine]. AB - The authors induced pulmonary silicosis in albino rats by the intratracheal method with 50 mg mixed middling slime from Panaguiriste and quartz dust, dissolved in I ml physiological solution. The animals received a single treatment and were killed on 90th day of the experiment. Some intime characteristics of collagen-forming mechanisms in lungs were followed up via routine histological, histochemical, enzymohistochemical/ acid phosphatase activity, beta glucoronidase, AS-esterase and lipase/ and autoradiographic/H3 labelled proline/ methods. It was established, that under the effect of both dusts from Panaguiriste mines studied/ mixed dust and pure quartz dust/ considerable deviations developed in the structure of the lungs of the experimental animals in the enzyme activity of acid phosphatase and carbon esterase, accompanied by various forms of fibrosis in the organ, depending on the dust applied. The difference in the composition of both dusts quartz and mixed dust slime - had an effect on the degree of the deviations, but not on their character, manifested in: I. specific protective reaction of organism, with activation of the ferments from the group of carbon esterase in macrophagic elements of interstitium and around the silicotic alterations in the lungs of the experimental animals; 2. inhibition of the maturation of macrophages in silicotic foci and 3. acceleration of the proliferation of fibroblasts with intensification of collagen-formation, accompanied by an enhanced activity of the ferments from the group of carbon esterase and in the cytoplasm of fibroblasts. The authors draw the conclusion that the complex use of routine histological, histochemical and quantitative isotope methods enables the more significant determination of fibrinogenicity of quartz-containing dust even at the early terms of their exposure under experimental conditions. PMID- 6294649 TI - [Role of chemotherapy in the treatment of Wilms' tumor in children]. AB - In the years 1962-1976, 168 children with nephroblastoma (Wilms Tumour) were treated in the Clinical Department of Child Oncology of the Institute of Mother and Child in Warsaw. In 150 cases, independently from surgery and radiotherapy, chemotherapy was performed. Our material was divided into 5 groups: a) without chemotherapy, b) chemotherapy with different drugs, c) chemotherapy with 1 course of Actinomycine D, d) chemotherapy with several courses of Actinomycine D, e) chemotherapy according to SIOP Trial. The results with reccurency free survival are as follows: a) 22,2%, b) 24,1%, c) 28,1%, d) 46,6%, e) 56,6%. In our material the best results were received with SIOP Trial chemotherapy. PMID- 6294650 TI - Odontodysplasia associated with unilateral malformations. A case report. PMID- 6294651 TI - The Rous sarcoma virus long terminal repeat is a strong promoter when introduced into a variety of eukaryotic cells by DNA-mediated transfection. AB - We characterized the transcriptional activity of the long terminal repeat (LTR) of Rous sarcoma virus by constructing a recombinant plasmid, pRSVcat, in which bacterial chloramphenicol acetyltransferase (CAT; acetyl-CoA:chloramphenicol 3-O acetyltransferase, EC 2.3.1.28) coding sequences are placed under LTR control. We find that the LTR directs relatively high levels of CAT synthesis within 48 hr after calcium phosphate-mediated introduction of this plasmid into CV-1 monkey kidney cells, chicken embryo fibroblasts, Chinese hamster ovary cells, HeLa cells, or mouse NIH/3T3 cells. The level of CAT synthesis is 3-fold higher in CV 1 cells and up to 10-fold higher in HeLa and mouse NIH/3T3 cells than after transfection with a related vector, pSV2cat, carrying CAT sequences under control of the simian virus 40 early promoter. We have shown, by primer extension, that the amounts of CAT-specific mRNAs encoded by pRSVcat and pSV2cat correlate with the levels of CAT enzyme activity. By both S1 nuclease mapping and primer extension, we have demonstrated that the start site for RNA transcription within the LTR of pRSVcat corresponds to previous mapping data. We estimated transfection efficiencies by monitoring immunofluorescence induced by a rhodamine labeled CAT antibody. Our results indicate that the Rous sarcoma virus LTR can direct synthesis of high levels of functional mRNA and has a wide expression range. The observed high transcriptional activity of the LTR is significant because it has been postulated that this LTR promotes activity of adjacent cellular oncogenes. PMID- 6294653 TI - Differential subcellular localization of in vivo-phosphorylated and nonphosphorylated middle-sized tumor antigen of polyoma virus and its relationship to middle-sized tumor antigen phosphorylating activity in vitro. AB - A small fraction of polyoma virus middle-sized tumor (T) antigen is phosphorylated in vivo, resulting in a small amount of phosphotyrosine and phosphothreonine and significantly larger amounts of phosphoserine. When infected cells are separated into nuclear, plasma membrane, and low-speed supernatant fractions, 80-95% of in vivo-phosphorylated middle-sized T antigen is localized to the plasma membrane fraction, while 25-50% of [35S]methionine-labeled middle sized T antigen is found in the nuclear fraction and the same amount is found in the plasma membrane fraction. Immunoprecipitated T antigens contain a protein kinase activity that phosphorylates middle-sized T antigen at tyrosine residues. Eighty to 90% of this activity is located in the plasma membrane fraction. When immunoprecipitated T antigens are treated with alkaline phosphatase, middle-sized T antigen-phosphorylating activity decreases as 32PO4 is lost from in vivo 32P labeled middle-sized T antigen. The possibility that in vivo-phosphorylated middle-sized T antigen located in the plasma membrane is an active tyrosine specific kinase is discussed. PMID- 6294652 TI - Insulin activates a tyrosine-specific protein kinase in extracts of 3T3-L1 adipocytes and human placenta. AB - Insulin activates a tyrosine-specific cAMP-independent protein kinase when added directly to detergent extracts of differentiated 3T3-L1 adipocytes and humal placental membranes. The kinase is also activated by antibody to the insulin receptor and, to a lesser extent, by proinsulin. It catalyzes the phosphorylation of the 92,000-dalton component of the insulin receptor, histone, and casein; in each case, tyrosine is the principal amino acid modified. Under the conditions used to activate the kinase, insulin does not affect the rate of dephosphorylation of the receptor or of histone. The insulin-activated kinase is copurified with the human placental insulin receptor until the final elution from insulin-Sepharose. It remains to be established whether the kinase and the insulin receptor are separate molecules. PMID- 6294654 TI - Complete nucleotide sequence of the neuraminidase gene of influenza B virus. AB - The complete nucleotide sequence of the neuraminidase gene of influenza virus B/Lee/40 was derived from a cloned cDNA copy of virion RNA segment 6 and its corresponding mRNA. The RNA segment contains 1,557 virus-specific nucleotides, and the protein encoded by the longest open reading frame has a total of 466 amino acids with a molecular weight of 51,721. As is the case with the influenza A virus neuraminidases, the deduced amino acid sequence of the influenza B protein includes a single hydrophobic region near the amino terminus which would be capable of spanning the lipid bilayer of the viral or cell membrane. There are four potential glycosylation sites in the protein, two of which are near the amino-terminal hydrophobic region. Comparisons of the nucleotide and amino acid sequences with those of influenza A virus neuraminidases revealed seven regions of extensive homology within the central portion of the molecules, including 12 conserved cysteine residues. Five other cysteine residues in the terminal portions were also conserved. PMID- 6294655 TI - Affinity labeling of a transforming growth factor receptor that does not interact with epidermal growth factor. AB - Membrane components that interact with epidermal growth factor (EGF) and transforming growth factors (TGFs) have been identified by covalent crosslinking to their respective 125I-labeled ligands. Under appropriate conditions, disuccinimidyl suberate or hydroxysuccinimidyl p-azidobenzoate cross-link receptor-bound 125I-labeled EGF to a 140- to 170-kilodalton (kDal) receptor species in membranes from both A431 human carcinoma cells and normal rat kidney cells. 125I-Labeled sarcoma growth factor (SGF), a TGF from virally transformed mouse 3T3 cells, also can be affinity-crosslinked to the 140- to 170-kDal EGF receptor species in membranes from A431 and rat kidney cells. The labeling of this receptor is inhibited when either excess unlabeled EGF or SGF is present during incubation of membranes with either 125I-labeled EGF or 125I-labeled SGF. In contrast, a second receptor species of 60 kDal is affinity-labeled with 125I labeled SGF but not with 125I-labeled EGF in membranes from both A431 and rat kidney cells. SGF and a TGF from virally transformed rat embryo cells inhibit the labeling of the 60-kDal species when present in excess during incubation of membranes with 125I-labeled SGF, whereas EGF is completely ineffective in inhibiting the labeling of this receptor. The data suggest that a specific 60 kDal receptor that displays high affinity for TGFs but not for EGF may mediate induction of the transformed phenotype. In addition, SGF and other TGFs interact with the 140- to 170-kDal EGF receptor that appears to mediate normal cell growth effects. PMID- 6294656 TI - Isolation and preliminary characterization of the GAL4 gene, a positive regulator of transcription in yeast. AB - The GAL4 locus encodes a positive regulator of the inducible galactose and melibiose genes of yeast. Using the yeast plasmid vector YEp13, we have cloned GAL4 by complementation of a gal4 mutation. Restriction endonuclease mapping of subclone DNA has delimited the region sufficient for complementation to a 3.2 kilobase segment of DNA. The GAL4 mRNA is 2.8 kilobases long, sufficient to encode a protein as large as 105,000 daltons. The concentration of the GAL4 transcript is about 0.1 per cell and is almost identical in galactose-induced and noninduced cells. This result is consistent with a previously proposed model in which the activity of the GAL4 protein and not the transcription of the GAL4 gene is modulated by galactose induction. PMID- 6294657 TI - Vitellogenin genes A1 and B1 are linked in the Xenopus laevis genome. AB - Genomic clones containing the Xenopus laevis vitellogenin gene B1 have been isolated from DNA libraries and characterized by heteroduplex mapping in the electron microscope, restriction endonuclease analysis, and in vitro transcription in a HeLa whole-cell extract. Sequences from the 3'-flanking region of the previously isolated A1 vitellogenin gene were found in the 5'-flanking region of this B1 gene. Thus, the two genes are linked, with 15.5 kilobase pairs of DNA between them. Their length is about 22 kilobase pairs (A1 gene) and 16.5 kilobase pairs (B1 gene) and they have the following arrangement: 5'-A1 gene spacer-B1 gene-3'. The analysis of heteroduplexes formed between the two genes revealed several regions of homology. Both genes are in the same orientation and, therefore, are transcribed from the same DNA strand. The possible events by which the vitellogenin gene family arose in Xenopus laevis are discussed. PMID- 6294658 TI - Cloning of developmentally regulated flagellin genes from Caulobacter crescentus via immunoprecipitation of polyribosomes. AB - Immunoprecipitation of Caulobacter crescentus polyribosomes with antiflagellin antibody provided RNA for the synthesis of cDNA probes that were used to identify three specific EcoRI restriction fragments (6.8, 10, and 22 kilobases) in genomic digests of Caulobacter DNA. The RNA was present only in polyribosomes isolated from a time interval in the Caulobacter cell cycle that was coincident with flagellin polypeptide synthesis. The structural gene for Mr 27,500 flagellin polypeptide was assigned to a region of the 10-kilobase EcoRI restriction fragment by DNA sequence analysis. Analysis of mutants defective in motility further established a correlation between the Mr 27,500 flagellin gene and the flaE gene locus [Johnson, R. C. & Ely, B. (1979) J. Bacteriol. 137, 627-634]. The other EcoRI fragments that hybridize with the immunoprecipitated polyribosome derived cDNA probe are also temporally regulated and have features that suggest they encode other polypeptides associated with the flagellum. Modifications were required to adapt the procedure of immunoprecipitation of polyribosomes for use with Caulobacter and should be applicable to the production of specific structural gene probes from other prokaryotic systems. PMID- 6294659 TI - Isolation and characterization of cDNA clones for human apolipoprotein A-I. AB - We have isolated cDNA clones encoding human apolipoprotein (apo) A-I. Twenty putative apo A-I cDNA clones were selected by screening 10,000 clones of an adult human liver cDNA library with an oligonucleotide probe. The probe was a mixture of synthetic 14-base-long DNA oligomers constructed to correspond to the codons for apo A-I amino acids 105-109. Four of these clones were examined further and showed 600- to 800-base-pair (bp) inserts. Preliminary restriction mapping and partial DNA sequence analysis indicated that the shorter inserts were a subset of the longer DNA inserts. DNA sequence analysis of the clone with an insert of approximately equal to 600 bp, designated pAI-113, revealed that it contained a DNA sequence corresponding to apo A-I amino acids 94-243. The DNA base sequence of this clone also contained a standard termination codon, polyadenylylation signal, and poly(A) tail. Partial DNA sequence of a second clone that contained an 800-bp insert, designated pAI-107, showed that it corresponded to apo A-I amino acids 18-243 and also included the 3' untranslated region. Isolation of these cDNA clones will facilitate molecular analyses of apolipoproteins in normal and disease states. PMID- 6294660 TI - Lateral mobility of cytochrome c on intact mitochondrial membranes as determined by fluorescence redistribution after photobleaching. AB - Lateral mobility of an active fluorescent derivative of cytochrome c on the membranes of giant mitochondria was measured by fluorescence redistribution after photobleaching. A diffusion coefficient of 1.6 X 10(-10) cm2/sec was determined for the labeled cytochrome c on inner mitochondrial membranes under conditions where succinate oxidase activity was demonstrated. This relatively low rate of diffusion, together with results of other investigators, is explained in terms of a model involving a dynamic equilibrium between freely diffusing and associated forms of electron-transfer components. PMID- 6294661 TI - Extensive intragenic sequence homology in two distinct rat lens gamma-crystallin cDNAs suggests duplications of a primordial gene. AB - The nucleotide sequences of two different rat lens gamma-crystallin cDNA clones, pRL gamma 2 and pRL gamma 3, have been determined. pRL gamma 3 contains the complete coding information for a gamma-crystallin of 173 amino acids whereas pRL gamma 2 is incomplete in that it lacks the codons for the first three amino acids of a separate but very homologous gamma-crystallin of identical length. Both rat gamma-crystallins are homologous to the known amino acid sequence of bovine gamma crystallin II which is only a single amino acid longer. The length of the region downstream the coding sequence to the A-A-T-A-A-A polyadenylylation signal sequence is 40 nucleotides in each clone. In pRL gamma 3 the poly(A) signal sequence is followed at 14 nucleotides by a remnant of the poly(A) tail which indicates that this clone contains a complete 3' noncoding region. pRL gamma 2 has only seven nucleotides following this signal sequence and no poly(A) tail, suggesting an incomplete 3' end. The cDNA clones show an overall nucleotide sequence homology of 85%. The mutual homology at the amino acid level is 73% whereas their amino acid homology with bovine gamma-crystallin II is about 70%. The nucleotide sequence of each clone also reveals a high intragenic homology and seems to be duplicated in itself. We suggest that the gamma-crystallin genes have arisen by multiple duplications of a primordial gene which consisted of about 120 nucleotides. PMID- 6294662 TI - Recovery of myc-specific sequences by a partially transformation-defective mutant of avian myelocytomatosis virus, MC29, correlates with the restoration of transforming activity. AB - Avian myelocytomatosis virus MC29 transforms fibroblasts and macrophages in vitro. Recently we isolated three deletion mutants of MC29 that have a decreased ability to transform macrophages while retaining their capacity to transform fibroblasts. One of these mutants, MC29 td10H, on passage through chicken embryo cultures gave rise to a recovered virus MC29 10H B1, which has regained the ability to transform macrophages efficiently. Immunoprecipitation analysis of MC29 10H B1-infected cells revealed a 108,000-dalton gag-myc polyprotein as opposed to the 90,000-dalton protein of MC29 td10H or the 110,000-dalton polyprotein of wtMC29. Tryptic peptide mapping studies demonstrated that the 108,000-dalton protein had acquired v-myc peptides that were lost from the td10H 90,000-dalton polyprotein and two novel peptides. Restriction enzyme analysis of the MC29 10H B1 proviral DNA also showed that myc sequences had been acquired. These results suggest that MC29 td10H has recombined with c-myc sequences to generate a recovered virus, MC29 10H B1. PMID- 6294663 TI - Hypolipidemic drugs are inhibitors of phosphatidylcholine synthesis. AB - Clofibric acid (CPIB) and several other systemic hypolipidemic drugs are shown to block phosphatidylcholine synthesis by inhibiting cholinephosphotransferase (ChoPTase; CDPcholine:1,2-diacylglycerol cholinephosphotransferase, EC 2.7.8.2) and particularly lysolecithin acyltransferase (LLAcylTase; acyl-CoA:1-acylglycero 3-phosphocholine O-acyltransferase, EC 2.3.1.23) of rat liver microsomes. Whereas millimolar drug concentrations are required to affect de novo lecithin synthesis catalyzed by ChoPTase, reacylation of lysolecithin by LLAcylTase is inhibited at micromolar levels. Increasing effectiveness in ChoPTase inhibition is observed in the series CPIB, SaH-42-348, tibric acid, S-321328, WY-14643, S-8527, and DH-990, with IC50 ranging from 22 mM (CPIB) to 0.3 mM (DH-990). LLAcylTase inhibition by the hypolipidemic drugs follows the same general pattern, but IC50 concentrations range from 9 mM (CPIB) to 40 microM (DH-990). The agents inhibit ChoPTase (Ki, 25 0.25 mM) and LLAcylTase (Ki, 10-0.025 mM) noncompetitively. The data suggest that inhibition of phosphatidylcholine synthesis, particularly by the LLAcylTase pathway, may be related to a drug's effectiveness in decreasing serum triglyceride and cholesterol levels by blocking lipoprotein synthesis. PMID- 6294664 TI - Human adult T-cell leukemia virus: molecular cloning of the provirus DNA and the unique terminal structure. AB - Adult T-cell leukemia virus (ATLV) is a human retrovirus closely associated with adult T-cell leukemia. The integrated provirus DNA and cDNA from virion RNA were molecularly cloned and their structures were analyzed. Clone lambda ATM-1 of an integrated provirus DNA in the MT-1 cell line, established from adult T-cell leukemia cells by cocultivation with cord lymphocytes, contained DNA about 13,000 base pairs (bp) long and long terminal repeats (LTR) at both ends of the viral sequence that were about 8,000 bp long. These two LTR sequences were linked to cellular sequences with direct repeats of 7 bp. Each LTR consisted of 754 bp including inverted repeats of 2 bp at the ends and the T-A-T-A-A box, characteristics in common with those of LTRs of other known retroviruses. Adjacent to the 5' LTR there was a sequence identical to the tRNAPro binding site in murine leukemia virus, suggesting that tRNAPro is a primer for reverse transcription of the viral genome. From these structural features, the mechanism of ATLV replication was suggested to be the same as that of other known animal retroviruses. However, the length of the small terminal repeats at the ends of the RNA genome, 228 +/- 1 bases, is much longer than the lengths, up to 80 bases, of those in avian, mouse, or primate retroviruses so far analyzed. These findings suggest that ATLV should be classified in a distinct group of retroviruses with bovine leukemia virus which also makes unusually long strong-stop cDNA. PMID- 6294665 TI - Nucleotide sequence requirements for specific initiation of transcription by RNA polymerase I. AB - The nucleotide sequence(s) specifying RNA polymerase I initiation has been investigated by studying the transcription of deleted and nondeleted mouse ribosomal RNA gene (rDNA) templates in vitro. The deletion of 5'-flanking sequences upstream from position -- 39 did not affect transcriptional activity, but removal of sequences between positions -- 39 and -- 34 resulted in a 90% decrease of rDNA transcription. The template activity was completely eliminated by the further deletion of nucleotides -- 33 to -- 13. It is concluded that sequences between -- 34 and -- 12, upstream from the transcribed region, represent an essential control region for the initiation of transcription in vitro. Therefore, this region may be functionally analogous to the T-A-T-A box of RNA polymerase II promoters. In addition to this control region, sequences located further upstream (between positions -- 45 and -- 169) may also exert some function in efficient transcription initiation as revealed by competition experiments between wildtype and mutant rDNA templates. PMID- 6294667 TI - Transfer of Bacillus thuringiensis plasmids coding for delta-endotoxin among strains of B. thuringiensis and B. cereus. AB - The recently discovered high-frequency transfer of plasmids between strains of Bacillus thuringiensis was used to study the genetic relationship between plasmids and production of the insecticidal delta-endotoxin crystal. Three strains of B. thuringiensis transmitted the Cry+ (crystal-producing) phenotype to Cry- (acrystalliferous) B. thuringiensis recipients. Agarose gel electrophoresis showed that one specific plasmid from each donor strain was always present in Cry+ "transcipients." The size of the transmissible crystal-coding plasmid varied with the donor strain, being 75 MDal (megadaltons) in size in HD-2, 50 MDal in HD 73, and 44 MDal in HD-263. Immunological analysis showed the Cry+ transcipients to be hybrid strains, having flagella of the recipient serotype and crystals of the donor serotype. These results demonstrate that the structural genes for the delta-endotoxin are plasmid borne. Crystal-coding plasmids also transferred into two strains of the related species Bacillus cereus and yielded transcipients that produced crystals of the same antigenicity as the donor strain. PMID- 6294666 TI - Analysis of desmin and vimentin phosphopeptides in cultured avian myogenic cells and their modulation by 8-bromo-adenosine 3',5'-cyclic monophosphate. AB - The intermediate filament proteins desmin and vimentin are two of the major 32P phosphate acceptors in chicken myotubes differentiating in tissue culture. Analysis of the desmin and vimentin phosphopeptides by two-dimensional tryptic peptide mapping shows that both proteins are phosphorylated at multiple sites, giving rise to 5 phosphopeptides in desmin and as many as 11 in vimentin. Addition of the cAMP analogue 8-bromoadenosine 3',5'-cyclic monophosphate (8 BrcAMP) to the culture medium of mature (8-day-old) myotubes results in a 2- to 3 fold increase in PO4 incorporation into desmin and vimentin. Two-dimensional tryptic analysis of desmin and vimentin from 8-BrcAMP-treated myotubes shows increased 32PO4 incorporation into a subset of the phosphopeptides observed in control cells. Comparison of phosphopeptides from the two proteins shows the presence of at least three comigrating peptides. All three comigrating peptides exhibit cAMP-dependent increases in 32PO4 incorporation in vimentin, while only two of the three exhibit 8-BrcAMP-dependent responses in desmin. While these peptides are the only two that are sensitive to 8-BrcAMP in desmin, vimentin contains additional peptides that exhibit increased 32PO4 incorporation in response to 8-BrcAMP. This result suggests the existence of both common and distinct phosphorylation sites between desmin and vimentin that may be differentially regulated by cAMP. Thus, desmin and vimentin, even though structurally related, may be capable of responding differently to physiological stimuli. PMID- 6294668 TI - Transcriptionally active DNA region that rearranges frequently in murine lymphoid tumors. AB - A DNA region not associated with conventional immunoglobulin gene rearrangement is rearranged in many lymphoid tumors. This region, designated here as lymphoid rearranging (LyR) DNA, was cloned from plasmacytoma J558 in which it had recombined 5' to a constant (C) region of the alpha heavy (H) chain gene, C alpha, within a switch (S) region, S alpha, involved in the switching of CH genes. Sequence determination established that LyR DNA had recombined within a S alpha recombination unit. LyR DNA does not originate from the H chain locus, and discordance between LyR DNA and CH copy number in certain lines suggests that LyR DNA probably derives from another chromosome. LyR DNA rearrangement is a characteristic of tumors of mature B cells; it was detected in 24 of 28 plasmacytomas and B-cell lymphomas, usually as LyR-S alpha, but not in 11 Abelson retrovirus-induced lymphomas of B-cell precursors nor detectably in normal B cells. In contrast, rearrangement was observed in only 3 of 18 T-cell lymphomas, and none of seven nonlymphoid lines. Most tumor lines (49 of 52), whether lymphoid or not, contained a low level of polyadenylylated LyR transcript(s), but several new RNA species with differences in their 5' regions appeared in B-cell lines in which LyR DNA was rearranged, suggesting that rearrangement may activate a new promoter or mode of splicing. The results suggest that the LyR-S alpha rearrangement represents a translocation to chromosome 12 that alters expression of LyR-encoded genes; hence, it may have participated in lymphoid tumor oncogenesis. PMID- 6294669 TI - Isolation of the yeast regulatory gene GAL4 and analysis of its dosage effects on the galactose/melibiose regulon. AB - GAL4 is a classically defined positive regulatory gene controlling the five inducible structural genes of galactose/melibiose utilization in yeast. The positive regulatory function of the GAL4 gene product in turn is controlled by the product of another gene, the negative regulator GAL80. We have cloned a 3.1 kilobase fragment containing GAL4 by homologous complementation using the multicopy chimeric vector YEp24 and demonstrated that multiple copies of GAL4 in yeast have pronounced dosage effects on the expression of the structural genes. Yeast transformed with GAL4-bearing plasmid become constitutive for expression of the galactose/melibiose genes, even in normally repressing (glucose) medium. Multiple copies of the GAL4 plasmid also increase expression of the structural genes in inducing (galactose) medium and can partially overcome the effects of a dominant super-repressor mutant, GAL80S. Using an internal deletion in GAL4, we have demonstrated that these dosage effects are due to overproduction of GAL4 positive regulatory product rather than an effect of the flanking sequences titrating out a negative regulator. These results point to the importance of competitive interplay between the positive and negative regulatory proteins in the control of this system. We have also used the dosage effect of GAL4 plasmid in combination with different GAL4 and GAL80 alleles to create new phenotypes. We interpret these phenotypes as indicating that (i) the repressing effects of glucose, at least in part, are mediated by the product of the negative regulatory gene, GAL80, and (ii) the GAL80 protein may have specific interactions with the control regions of the structural genes. PMID- 6294670 TI - Preparation and characterization of a pentaammineruthenium(III) derivative of horse heart ferricytochrome c. AB - A stable complex between pentaammineruthenium(III) and histidine-33 in horse heart ferricytochrome c is formed in the reaction between aquopentaammineruthenium(II) and the protein at pH 7. HPLC of the tryptic hydrolysate of the modified protein was employed to identify the pentaammineruthenium binding site. Spectroscopic measurements show that the integrity of the native structure in the vicinity of the heme c group is maintained in the ruthenium-modified protein. The reduction potentials are: heme c (Fe3+/2+), 0.26 V; Ru(NH3)5(His-33)3+/2+, 0.15 V (vs. normal hydrogen electrode). PMID- 6294671 TI - The assessment of a new protein source--rapeseed. PMID- 6294673 TI - Golgi complex function in the excretion of renal kallikrein (41502). PMID- 6294672 TI - Demonstration of alpha 2-adrenergic receptors in rat pancreatic islets using radioligand binding (41498). PMID- 6294674 TI - Reserpine inhibits rat anterior pituitary hormone secretion in vitro: effects on GH, TSH, and LH. PMID- 6294675 TI - Discriminative stimulus properties of d-amphetamine in pigeons. AB - Two out of four pigeons were successfully trained in an operant procedure to discriminate between the presence and absence of the effects induced by d amphetamine (final dose: 1.6 mg/kg). The solvent (saline) or d-amphetamine was administered intramuscularly (IM) 30 min prior to training. Tests with other drugs and dosages indicated that l-amphetamine (ED50 = 0.55 mg/kg) and cocaine (ED50 = 1.05 mg/kg) fully generalized to d-amphetamine (ED50 = 0.35 mg/kg), whereas drugs such as p-hydroxy-amphetamine (1.6 and 3.2 mg/kg), morphine (1.5, 3.0 and, 6.0 mg/kg), and delta 9-THC (0.125, 0.25, and 0.50 mg/kg) failed to do so at the doses tested. Apomorphine (0.25 and 0.50 mg/kg) and LSD-25 (0.04 and 0.08 mg/kg) produced intermediate results. Pretreatment with haloperidol (dose range: 0.04 to 1.28 mg/kg), but not propranolol (10 and 20 mg/kg), attenuated significantly the d-amphetamine (1.6 mg/kg) stimulus effects. The two pigeons emitted predominantly d-amphetamine appropriate responses when the training dose (1.6 mg/kg) of d-amphetamine was tested on different occasions 15, 60, and 120 min after the administrations. One bird emitted mostly vehicle appropriate responses when tested 240 min after the d-amphetamine injection whereas the other bird performed d-amphetamine appropriate responses. Selection of the non-drug associated key occurred in the two birds when testing was carried out 480 min (8 hrs) after the administration of d-amphetamine. PMID- 6294676 TI - Brain cyclic AMP and memory in mice. AB - A phosphodiesterase inhibitor 4-(3-cyclopentyloxy-4-methoxyphenyl)-2-pyrrolidone (Rolipram, 10 mg/kg IP) administered immediately, but not 3 hr post-training, reversed an amnesia for an inhibitory avoidance response induced by the protein synthesis inhibitor anisomycin. Immediate post-training administration of Rolipram also enhanced retention for a weakly learned avoidance response. Unshocked animals did not show increased test latencies thus ruling out conditioned aversion as an explanation for the enhanced avoidance. Mice treated with Rolipram (10 mg/kg after training showed elevated cyclic AMP but not cyclic GMP in frontal cortex, thalamus, and hypothalamus. These results support the suggestion that cyclic AMP may play a role in memory processes. PMID- 6294677 TI - Attenuation of pentobarbital-elicited hypothermia in rats with a history of pentobarbital-LiCl pairings. AB - Rats were given five separate pairings (sequential IP injections) of pentobarbital and lithium chloride, both hypothermia-inducing agents. When the animals were subsequently tested with a single injection of pentobarbital alone, they exhibited an attenuated hypothermia relative to controls that had either (a) received pentobarbital-LiCl pairings spaced twenty-four hours apart, or (b) received only placebo injections of normal saline. This phenomenon provides further evidence that rats can learn an association between drug states and may help to explain why pentobarbital-LiCl pairings tend to eliminate pentobarbital's capacity to produce a conditioned flavor aversion. PMID- 6294678 TI - Chronic phencyclidine treatment decreases phencyclidine and dopamine receptors in rat brain. AB - Chronic phencyclidine treatment (10 mg/kg/day, SC for 14 days) significantly decreased the number of [3H]phencyclidine and [3H]spiperone binding sites in rat brain. [3H]Dihydromorphine binding was not affected by the same treatment. An acute treatment with phencyclidine (10 mg/kg, SC) did not modify any of the binding sites under study. These results suggest that a chronic phencyclidine treatment induces a down-regulation of phencyclidine and dopamine receptors without affecting opiate receptors. These reductions in the number of phencyclidine and dopamine binding sites might be related to the development of tolerance and/or dependence to phencyclidine. PMID- 6294679 TI - Similarities of the cataleptic state induced by beta-endorphin and morphine. AB - A variety of behavioral tests were used to characterize the cataleptic state induced by various treatments. Besides catalepsy, posture, locomotion, rigidity and the presence of reflexive responses were assessed. Measures of analgesia and body temperature were taken. The behavioral profiles of beta-endorphin, morphine, etonitazene, haloperidol, arecoline and GABA were compared at the time maximal catalepsy scores were obtained. Results indicated that, for an equivalent degree of catalepsy, the profile of beta-endorphin was similar to that of opiates, except for changes in body temperature; beta-endorphin's profile differed markedly from that of haloperidol, arecoline and GABA. Catalepsy was less pronounced with the latter two drugs. There were similarities in the behavioral profile of haloperidol and arecoline. PMID- 6294680 TI - Generalization between benzodiazepine- and triazolopyridazine-elicited discriminative cues. AB - Using a milk reinforced two-lever operant procedure, rats were trained to discriminate 3 mg/kg chlordiazepoxide (CDP) from saline. Following this, generalization experiments were conducted with the triazolopyridazine CL 218,872, a synthetic non-benzodiazepine (BDZ) ligand for the BDZ receptor. CL 218,872 produced CDP lever selection in a dose related fashion and thus generalized to the standard CDP treatment. However, this generalization was antagonized by the concurrent administration of pentylenetetrazol or amphetamine, but not by strychnine or bicuculline. Also, there was evidence for cross tolerance for a sedative effect between CDP and CL 218,872. PMID- 6294681 TI - Effects of the combination of tripelennamine and pentazocine at the behavioral and molecular levels. AB - The purpose of the present experiments was to determine if the antihistamine tripelennamine potentiates the morphine-like effects of the narcotic-antagonist analgesic pentazocine at the behavioral level or the molecular level or both. At the behavioral level, the effects of pentazocine were determined alone and in combination with tripelennamine in rats trained to discriminate between saline and either morphine or the psychotomimetic narcotic derivative SKF 10,047. The interaction between pentazocine and tripelennamine were also evaluated in the guinea-pig ileum preparation and in the [3H]-naloxone opiate receptor binding assay. Tripelennamine significantly enhanced the morphine-like discriminative stimulus effects of pentazocine and markedly reduced the SKF 10,047-like stimulus effects of pentazocine. Naloxone antagonized the morphine-like effects of pentazocine plus tripelennamine. Pentazocine significantly constricted pupils in the rat, an effect which was additive with the mydriatic effects of tripelennamine. Inhibition of the twitch-height of the electrically stimulated guinea-pig ileum by pentazocine was not affected by tripelennamine. Further, tripelennamine did not modify the Ke for naloxone in antagonizing pentazocine. Inhibition of specific [3H]-naloxone binding by pentazocine was also not affected by tripelennamine. These results are consistent with the hypothesis that the potentiation of the morphine-like effects of pentazocine by tripelennamine which was observed behaviorally was not due to molecular interactions at the morphine receptor. At least a part of this interaction may be attributable to tripelennamine decreasing the psychotomimetic actions of pentazocine. PMID- 6294682 TI - Extent and control of shock affects naltrexone sensitivity of stress-induced analgesia and reactivity to morphine. AB - Opioid and nonopioid mediated changes in pain sensitivity have been observed after exposure to various stressful conditions. A series of inescapable shocks sequentially produces an early form of analgesia which is not affected by the opiate antagonist, naltrexone, and a late antinociceptive response which is sensitive to reversal by naltrexone. Here, this is shown to be true over a wide range of doses. In a further experiment subjects given either escapable or inescapable shock were analgesic immediately after the stress session. However, the analgesia of inescapably shocked subjects was more sensitive to reversal by naltrexone. A final experiment revealed that inescapably shocked subjects, but not escapably shocked subjects, were hyperreactive to the analgesic effects of morphine 24 hr after shock. These results suggest that activation of an opiate system occurs only after extended exposure to stress and that this activation is greater when the stress is inescapable. Implications for opioid versus nonopioid mechanisms of stress-induced analgesia are discussed. PMID- 6294683 TI - Failure to find electrophysiological correlates of chronic neuroleptic-induced oral dyskinesias in cats: somatosensory and substantia nigra evoked potentials, electroencephalogram, and caudate spindles. AB - Each of nine cats was prepared with 30 chronically implanted stainless steel gross electrodes in cortex, basal ganglia and other brain structures. Measurements were taken for 0.5-11.5 months of baseline, chronic daily administration of chlorpromazine, and withdrawal. In most cases there was also a second cycle of drug administration and withdrawal. Although we observed dramatic, persistent increases in licking behavior, suggestive of tardive dyskinesia, consistent correlated patterns were not observed in somatosensory or substantia nigra evoked potentials, electroencephalogram, or spindling evoked in cortex by caudate stimulation. PMID- 6294684 TI - Cyclic AMP and cyclic GMP response to stress in brain and pituitary: stress elevates pituitary cyclic AMP. AB - Male rats were exposed to six stressors (saline injection, cold, forced running, Formalin injection, immobilization, electric footshock) for 15, 30, or 60 min. Following sacrifice by microwave irradiation, cyclic AMP and cyclic GMP levels were measured in pituitary, pineal and 8 regions of rat brain. All stressors except saline increased plasma corticosterone, plasma prolactin and pituitary cyclic AMP levels compared to control animals. The magnitude of the pituitary cyclic AMP response was highly correlated with the intensity of the stress as determined by the levels of plasma prolactin. Electric footshock increased pituitary cyclic AMP levels over 10 fold and plasma prolactin over 60 fold. Cyclic AMP levels in other brain regions were not altered. Cerebellar cyclic GMP was increased only by stressors that involved increased motor activity. PMID- 6294685 TI - Naloxone and play fighting in juvenile rats. AB - According to the opioid hypothesis of social attachment opiate receptor blockade should increase the need for social contact. Yet naloxone reduces play fighting [11], a major form of social interaction in young rats. This observation might be reconciled with the opioid attachment hypothesis if it could be shown that naloxone produced compensatory increases in other social activities or if naloxone shortened play fighting bouts without reducing their frequency. In the present experiment naloxone reduced play fighting in a dose-dependent fashion. However, the frequency of play bouts was reduced, their duration unchanged and no compensatory increase in social sniffing and grooming was observed. In addition, naloxone inhibited rearing almost as potently as it affected play fighting. PMID- 6294686 TI - Enhancement of osmotic- and hypovolemic-induced drinking by chlordiazepoxide in rats is blocked by naltrexone. AB - Recent reports indicate that benzodiazepine-induced hyperphagia can be antagonised by naloxone, an opiate antagonist. Benzodiazepines are also known to facilitate water ingestion in water-deprived rats, and the present study showed that in addition, benzodiazepine treatment can enhance drinking which is elicited by an osmotic thirst stimulus (2 M hypertonic saline) or by a hypovolemic thirst stimulus (20% polyethylene glycol). In both cases, low dose levels of naltrexone (also an opiate antagonist) dose-dependently suppressed the facilitation of thirst-aroused drinking by chlordiazepoxide. Taken with recent biochemical data these behavioral results indicate that the enhancement of ingestive responses by benzodiazepines may depend upon a naloxone-reversible release of endogenous opioid peptides. PMID- 6294687 TI - Effects of prenatal and postnatal exposure of rats to alcohol: changes in (Na+ K+) ATPase. AB - Maternal ethanol consumption produces a reduction in postnatal growth. We have studied especially changes of liver and brain. This reduction is more marked if the alcoholic offspring are maintained with their biological mothers than if they are kept with surrogate mothers. Rats exposed prenatally to alcohol show a marked accumulation of fat in the liver and a significant proliferation of liver endoplasmic reticulum. No change in the postnatal development of liver alcohol (ADH) and acetaldehyde dehydrogenases (ALDH) (high and low Km) is observed in offspring from alcoholic mothers, with the exception of slightly higher ALDH (low Km) for the offspring that remain with alcoholic mothers. The postnatal development of the liver (Na+-K+) ATPase is also similar in control and alcoholic groups. However, in the case of the enzyme from the brain, a lower ATPase activity is observed in the group derived from alcoholic mothers. Interestingly, at 20 days of postnatal period, an induction of the ATPase (from liver and brain) was observed when the group of offspring from alcoholic mothers were kept on an alcohol diet. PMID- 6294688 TI - Lesions of cholinergic forebrain nuclei: changes in avoidance behavior and scopolamine actions. AB - The acquisition of active (shuttle-box) and passive avoidance conditioned responses and the effects of scopolamine on acetylcholine (ACh) output in freely moving rats and on conditioned responses were investigated 20 days after placing a unilateral lesion in the magnocellular forebrain nuclei (MFN). In the lesioned rats spontaneous ACh output from the cerebral cortex ipsilateral to the lesion was slightly decreased, while on the other hand the increase in ACh output elicited by scopolamine was strongly reduced. Sham operated rats always performed more active avoidance responses than MFN lesioned rats in the daily training shuttle-box sessions, and the facilitating effect of scopolamine (1 mg/kg IP) on the shuttle-box performance was suppressed. However the lesion did not disrupt the shuttle-box performance whenever training had taken place before the lesion. In the lesioned rats retested 30 min after the training trial, an impairment of the passive avoidance response was found. The effect of the lesion was potentiated by scopolamine. The results show therefore that MFN lesions impair the cortical cholinergic mechanisms, whose activity seems to play an important role in cognitive functions. PMID- 6294689 TI - Dopaminergic properties of nomifensine. AB - Nomifensine and a proposed dihydroxy metabolite produced stimulation of motor behavior in mice with nomifensine being more potent. Weak cage-climbing behavior (stereotypy) was also produced. The stimulatory effects were greater in mice in which dopamine receptor sensitivity was increased by long-term haloperidol. Both of the analogs were potent inhibitors of dopamine and norepinephrine uptake in vitro with nomifensine approximately 3 times more potent than the metabolite. In contrast, the two analogs had weak affinity for the post-synaptic dopamine receptor (as measured by displacement of ligand binding in vitro) with dihydroxynomifensine approximately 6 times more potent than nomifensine. These results suggest that the behavioral effects of nomifensine are largely dependent on presynaptic catecholamine mechanisms but that weak direct dopamine agonist properties do exist, particularly in vivo where the drug may be metabolized to a more active form. PMID- 6294690 TI - Intra-raphe muscimol induced hyperactivity depends on ascending serotonin projections. AB - Acute microinjections of the GABA agonist, muscimol (100 ng), into either the dorsal (DR) or the median (MR) raphe nucleus of etherized rats induced post anesthesia hyperactivity as measured in photocell chambers. The increased activity counts seen after MR injections, furthermore, were 4 times greater than those following DR injections. In animals implanted with chronically indwelling cannulae, a muscimol (25-400 ng) dose-response analysis confirmed the greater sensitivity of the MR site. Subsequent experiments thus employed only MR cannulae. The benzodiazepine, chlordiazepoxide, in a subataxic dose (3.8 mg/kg, IP) by itself did not affect activity level, but enhanced the locomotor response to low doses (25-50 ng) of muscimol. Conversely, a sub-convulsant dose of the GABA antagonist, bicuculline (1.1 mg/kg, IP), completely blocked the hyperactivity produced by muscimol (50-100 ng). Bilateral electrolytic destruction of the ventral tegmental nuclei of Gudden produced hyperactivity, but failed to alter the effect of muscimol. Forebrain 5-hydroxytryptamine (5-HT, serotonin) depletion following administration of 5,7-dihydroxytryptamine did not affect baseline activity level, but markedly attenuated the locomotor response produced by intra-MR injections of muscimol. These data suggest that midbrain GABA neurons modulate activity level through a direct action on 5-HT neurons, and indicate that intra-MR muscimol induced hyperactivity depends on intact ascending 5-HT fibers. PMID- 6294691 TI - Effects of dithiocarb and (+)-catechin on the glutathione-conjugating system in rat liver cytosol in vivo and in vitro. PMID- 6294692 TI - Drug interactions at (pharmacodynamically active) receptor sites. PMID- 6294693 TI - The cellular resting and action potentials: interpretation based on the association-induction hypothesis. AB - The Hodgkin, Huxley, and Katz theories of resting and action potentials are based on the membrane theory, which holds that cell K+ and water exist in the free state. Reviewed here are these theories of cellular potential along with the results of experimental testings. Reviewed also is Ling's association-induction (AI) hypothesis, which holds that all K+ is absorbed selectively and singly on anionic protein sites and that cell water is absorbed in multilayers on extended chains of "matrix proteins." In the development of the AI model, molecular mechanisms of cell permeation and electric potentials were presented according to which the potentials are surface-adsorption phenomena. Thus they resemble those suggested by Baur rather than the membrane potentials proposed by Ostwald and Bernstein. In the present review it is shown that the AI version of the surface adsorption model can account for evidence supporting the Hodgkin, Huxley, Katz approach as well as evidence against it-including extensive recent confirmation of the absorbed state of K+ in muscle. PMID- 6294694 TI - Oxidation of tyrosine to dopachrome by peroxidase isolated from murine melanoma. AB - Peroxidase, isolated from B16 mouse melanoma, converted tyrosine to dopachrome in the presence of either dopa or dihydroxyfumarate co-factor. A suspended homogenate of cloned, cultured B16 mouse melanoma cells also showed peroxidatic conversion of tyrosine to dopachrome in the presence of dihydroxyfumarate co factor. The findings confirm previous histochemical, autoradiographic histochemical, and EM-histochemical studies showing that melanoma peroxidase can convert tyrosine to melanin. PMID- 6294695 TI - Flavor-toxicosis associations: tests of three hypotheses of long delay learning. AB - Rats were prepared with gastric cannulas and esophageal fistulas to assess the contribution of taste and smell, postingestinal stimuli, and the combination of the two in forming flavor-toxicosis associations. Groups were either given sugar water to drink, a stomach intubation of sugar water, both sugar water to drink and a stomach intubation of sugar water, or not given sugar water. The experimental and pseudo-conditioning control animals were given a LiCl injection and the control animals were given a NaCl injection one hour later. They were tested for drinking of sugar water the next day. It was found that taste and smell are the primary stimuli for forming food aversions. Furthermore, postingestional stimuli are not sufficient to produce this type of learning. PMID- 6294696 TI - Pre-test epinephrine injections reverse DDC-induced retrograde amnesia. AB - The effects of pre-test systemic administration of epinephrine on DDC-induced retrograde amnesia (RA) for discriminated Pavlovian fear-conditioning were examined in rats. Epinephrine reversed RA with the optimal dosage being 0.05 mg/kg. Apparently, the effect was specifically reversal of amnesia since (a) performance was restored to control levels, but no higher, and (b) sensitization and activity-related artifacts were minimized. These results are consistent with those showing reversal of RA by pre-test administration of hormones or catecholamine agonists. That is, they suggest that amnesia is due to a retrieval deficit rather than to failure of memory storage. Results are discussed in terms of epinephrine-induced modulation of storage and retrieval processes through central and peripheral mechanisms. PMID- 6294697 TI - Unique contribution of catecholamine receptors in the brain of the cat underlying feeding. AB - After a Collison cannula was implanted bilaterally in the cat for intracerebroventricular (ICV) injection, individual patterns of food intake were determined. A mixture of dry and canned cat food was provided to the animal every morning at the same time for 1.0 hr. Once intakes had stabilized, either norepinephrine (NE) or an adrenoreceptor blocking agent was infused in one lateral ventricle by gravity flow over an interval of 30-60 sec. NE given ICV in a dose of 25-75 micrograms 15 min before the period of feeding enhanced both the amount of food consumed by the cat and the duration of its feeding for up to 1.0 hr. When phentolamine in a dose of 25-100 micrograms was infused 15 min prior to the feeding interval, the cat's intake of food was attenuated in a dose-dependent manner, again in terms of both the quantity consumed and the interval of eating. Propranolol was essentially without effect on food intake. In the fully satiated cat, NE infused ICV 15-45 min following the termination of its morning feeding episode tended to augment food intake at the highest 100 micrograms dose, but the effect was not significant. The dose-dependent hypothermia typically observed after NE is given ICV to the cat was found to be entirely independent of the feeding response. Overall, these findings show that noradrenergic pathways in the cat's feeding system apparently bias or otherwise modulate the amount of food consumed. As in other species, the feeding mechanism appears to be mediated through alpha-adrenergic receptors presumed to be in the diencephalon of the cat. PMID- 6294698 TI - Vagal mediation of the cholecystokinin satiety effect in rats. AB - Central (intracerebroventricular) and peripheral (intraperitoneal) injections of the octapeptide of cholecystokinin (CCK-8) were compared to determine the most effective route of administration to elicit satiety for food intake in the rat. Subdiaphragmatic bilateral vagotomy and spinal cordotomy (T2-T3) were also performed to investigate the importance of visceral nerves for the satiety effect. CCK-8 suppressed feeding and elicited satiety resting behavior when injected peripherally but it was less effective when injected centrally. The satiety effect of CCK-8 or CCK-33 following peripheral injections was blocked by vagotomy whereas spinal cordotomy had no effect. The results indicate that some component of the vagus is required to mediate the peripherally induced cholecystokinin satiety effect, but the splanchnic nerves are not necessary. The weak effect of CCK-8 following ventricular administration is additional evidence suggesting that cholecystokinin of intestinal origin acts in the periphery rather than directly on the brain to elicit its typically rapid satiety effect in rats. PMID- 6294699 TI - C-terminal octapeptide of cholecystokinin decreases food intake in obese men. AB - Six of eight obese men ate significantly less food during an intravenous infusion of the C-terminal octapeptide of cholecystokinin (CCK-8, 4 ng . kg-1 . min-1) than during a saline infusion in a double blind experimental paradigm. Subjects stopped eating sooner during CCK-8. CCK-8 did not change the rate of eating. No overt side effects were reported or observed. This is the first report of the satiety effect of CCK-8 in obese humans and it suggests that the therapeutic potential of CCK-8 for the treatment of obesity deserves investigation. PMID- 6294700 TI - Disruption of conditioned taste aversion by the combined effects of LiCl and ECS. AB - Rats were taught a conditioned taste aversion by allowing them to drink sucrose (CS) for 5 min and 30 min later poisoning them with LiCl (UCS). Experimental animals were given ECS (80 mA for 250 msec) at 0, 15, 20, 25, 27.5, 30, 32.5, 35, 40, or 45 min after the CS. Only animals given ECS both within the CS-UCS interval and in close temporal proximity to the UCS (within 5 min) showed a significant, although limited, disruption of learning. At least two explanations are possible. The first is that apart from its toxicity, LiCl may also possess amnesic properties which interact with those of the ECS. Alternatively, ECS may have impaired the ability of the animals to fully experience the lithium-induced illness. PMID- 6294701 TI - Construction and characterization of a chimeric plasmid in Bacillus subtilis. PMID- 6294702 TI - Multimerization and replication of plasmid pBP11. PMID- 6294703 TI - Expression of cloned chloroplast DNA from Euglena gracilis in an in vitro DNA dependent transcription-translation system prepared from E. coli. PMID- 6294704 TI - Tn7 insertion mutations affecting the host range of the promiscuous IncP-1 plasmid R18. PMID- 6294705 TI - Genetic isolation and physical characterization of pAgK84, the plasmid responsible for agrocin 84 production. PMID- 6294706 TI - The kanamycin resistance transposon Tn2680 derived from the R plasmid Rts1 and carried by phage P1Km has flanking 0.8-kb-long direct repeats. PMID- 6294707 TI - Cloning of the replication and incompatibility regions of a plasmid derived from Rts1. PMID- 6294708 TI - Analysis of the Inc P-1 group plasmids R906 and R751 and their relationship to RP1. PMID- 6294709 TI - Characterization of the transposons Tn1822 (Tc) and Tn1824 (TpSm) and the light they throw on the natural spread of resistance genes. PMID- 6294710 TI - Mitochondrially encoded resistance to paromomycin in Saccharomyces cerevisiae: reinvestigation of a controversy. PMID- 6294711 TI - Complementation of transposition of tnpA mutants of Tn3, Tn21, Tn501, and Tn1721. PMID- 6294712 TI - Small circular DNA complexes in eucaryotic cells. PMID- 6294713 TI - Incompatibility group Y member relationships: pIP231 and plasmid prophages P1 and P7. PMID- 6294714 TI - Inhibition of cyclic AMP phosphodiesterase by flavonoids. PMID- 6294715 TI - Two cases of nevoid basal cell carcinoma syndrome. AB - Motivated by the diagnosis and treatment of two cases of nevoid or Goltz-Gorlin syndrome, we have taken a brief look at the literature and present these two cases, which display the four principal features defining this syndrome: multiple basal cell carcinomas, maxillary cysts, skeletal anomalies, and ectopic calcifications. Certain aspects are emphasized, including the association in one of them of basal cell carcinomas with a cystic adenoma or Brooke's tumor, which, although described by other authors, is infrequent. PMID- 6294716 TI - Regulation and adaptation of central norepinephrine receptor systems: modification by antidepressant treatments. PMID- 6294717 TI - Effect of morphine, ACTH, epinephrine, Met-, Leu- and des-Tyr-Met-enkephalin on beta-endorphin-like immunoreactivity of rat brain. AB - Morphine (1,0 mg/kg), ACTH1-24 (10.0 micrograms/kg), epinephrine (12.0 micrograms/kg), Met-enkephalin (2.0 and 5.0 micrograms/kg), Leu-enkephalin (2.0 micrograms/kg) and des-Tyr-Met-enkephalin (2.0 micrograms/kg) all produced marked reductions of beta-endorphin-like immunoreactivity in the rat diencephalon. At a dose of 0.4 mg/kg, naloxone had no effect of its own and was unable to reverse the depleting effect of the other substances. The depletion of beta-endorphin like immunoreactivity caused by the various treatments is attributable to release and subsequent degradation of beta-endorphin and/or of its precursors. The various behavioral effects of morphine, ACTH, epinephrine and the enkephalins may be explained by the release of endogenous beta-endorphin. PMID- 6294718 TI - Effects of gonadectomy and testosterone replacement on growth hormone response to alpha 2 adrenergic stimulation in the male rat. AB - The growth hormone (GH) response to clonidine in reserpine-pretreated rats is a putative in vivo model to reflect activation of central postsynaptic alpha 2 receptors. In the present study the influence of testosterone on the responsiveness of central alpha 2 receptors was investigated using this method. One week after operation the GH response to clonidine was drastically reduced in gonadectomized adult male rats compared to sham-operated controls. Testosterone replacement completely antagonized the effect. The results suggest an influence of testosterone on central postsynaptic alpha 2 receptors or on structures connected to these receptors. PMID- 6294719 TI - Lithium does not prevent ECS-induced decreases in beta-adrenergic receptors. AB - Electroconvulsive shock (ECS) reduces the number of rat cortical beta-adrenergic receptors. Lithium has been reported in several systems to prevent receptor changes induced by other agents. However, the present experiment reports that chronic lithium does not prevent the reduction in dihydroalprenolol binding induced by ten daily ECS treatments. PMID- 6294720 TI - Sensitivity amplification in biochemical systems. PMID- 6294721 TI - Relationship of enhanced survival during confluent holding recovery in ultraviolet-irradiated human and mouse cells to chromosome aberrations, sister chromatid exchanges, and DNA repair. PMID- 6294723 TI - [Enzymatic and structural aspects of activation of repair synthesis of DNA in mammalian chromatin]. PMID- 6294722 TI - A comparison of the DNA and chromosome repair kinetics after gamma irradiation. PMID- 6294724 TI - [Effect of adeturon on the survival and the blood system of mice under the effect of ionizing radiation of different types]. PMID- 6294725 TI - [Toxicity and radiation-protective effect of multi-component combinations of sulfur-containing substances]. PMID- 6294726 TI - Investigation on the distribution of 14C-triptophan and the incorporation into the central nervous system in rats pretreated with radioprotector and exposed to irradiation. PMID- 6294728 TI - [Diagnostic importance of 99mTc pertechnetate profiles in juvenile chronic arthritis]. PMID- 6294729 TI - [Lymphographic studies in the maxillofacial region and neck by injection of radiocolloid into the upper lip mucosa]. PMID- 6294727 TI - The colitides. AB - Diverse signs may be seen on barium enema in the various forms of colitis. Barium enema is only reasonably specific in the diagnosis of Crohn's disease and ulcerative colitis once the other possible diseases have been excluded from consideration by biopsy, microscopy, culture, or therapeutic trial. In particular, campylobacter colitis has proved to be such a common entity that many patients originally thought to have had one attack of ulcerative colitis, ischemic colitis, or Crohn's colitis may never have had these diseases. Many of the relatively specific signs can be seen in a variety of conditions. For example, aphthae in the colon may be seen in Crohn's disease, yersinia enterocolitis, Behcet's syndrome, amebiasis, ischemia, tuberculosis, and campylobacter colitis. Continuity of disease is characteristic in ulcerative colitis. Nevertheless, patches of healing may occur, so that a rare patient may be seen during a resolving attack of ulcerative colitis in whom only scattered patches of active disease can be seen on barium enema. In general, few radiologic signs on barium enema are truly specific for one disease, with penumatosis cystoides coli being an exception to this rule. However, the sensitivity of barium enema with currently available materials for double-contrast techniques is such that radiology continues to be useful at present in diagnosing the colitides and in managing patients. PMID- 6294730 TI - [Computer tomographic guided fine needle puncture of abdominal masses]. AB - CT-guided fine-needle aspiration biopsy was carried out in 177 patients with proven intra-, retro- or extraperitoneal lesions. This method verified the diagnosis in 89/132 cases (66%) with malignant neoplasia, metastases and recurrent malignant tumors. In 45 patients with benign or inflammatory lesions of the intra- and retroperitoneal space, fine-needle puncture showed a purulent liquid, inflammatory altered cell elements or a positive bacteriological result. The success of CT fine-needle puncture depends mainly on the examiner's experience, the exact localization of the needle, and careful handling of the cytological material. Fine-needle puncture should be applied simultaneously with CT when an undefined space-occupying lesion is present. Unsuccessful sonography is an indication for CT-guided fine-needle puncture. PMID- 6294731 TI - [Ankylosing spondylitis in women: comparison of scintigraphic results with clinical and roentgenologic findings]. AB - Since ankylosing spondylitis (S.a.) tends to be milder and less progressive in women than in men, there are great difficulties in diagnosing S.a. in female patients. The role of scintiscanning for investigation of S.a. in women is discussed by few examiners only, especially there are no investigations of quantitative sacroiliac scintigraphy in large groups of female patients. Quantitative sacroiliac scintigraphy was performed in 19 female patients with S.a. and in 30 control subjects. The sacroiliac/sacrum ratio (Index ISG/sacrum) was calculated. Significantly increased uptakes were found in the patient group compared with the controls. Especially in the early periods of S.a. the activity index was highest. When history and physical examination lead to a suspicion of S.a. and there is no verification by roentgenographic studies, sacroiliac joint scintigraphy is indicated. If clinical and laboratory findings are respected in each patient, quantitative sacroiliac scintigraphy is useful for the early diagnosis of S.a. in female patients. PMID- 6294732 TI - Studies of gallbladder contraction using intramuscular sincalide. AB - Intramuscular sincalide (the carboxy terminal octapeptide of cholecystokinin) was evaluated as an agent for producing gallbladder contraction. Following oral cholecystography 35 patients received intramuscular sincalide in one of two dosages, 18 patients received intravenous sincalide, and six patients were given intramuscular placebo. Generalized symptoms, often similar to those noted clinically, were more common following intravenous injection of sincalide; gallbladder contraction was greater following intramuscular injection. Both of these findings may be related to lower but more sustained blood levels of sincalide following intramuscular administration. Intramuscular sincalide may be useful for further studies of the value of cholecystokinin cholecystography. PMID- 6294734 TI - Spontaneous pneumothorax in primary cavitating lung carcinoma. AB - Primary cavitating lung carcinomas have not previously been described with spontaneous pneumothorax as the presenting manifestation. Two patients whose pneumothoraces resulted in the initial diagnosis of a primary cavitary lung carcinoma are described. Pathologic evidence of vascular invasion suggests tumor necrosis with secondary rupture into the pleural space as a likely mechanism for pneumothorax. PMID- 6294733 TI - Computed tomographic study of hormone-secreting microadenomas. AB - A review was made of the computed tomographic (CT) studies of 33 patients with hormone-secreting microadenomas that had been verified by transsphenoidal surgery and endocrinologic evaluation. In previous studies in small series of patients, the CT appearance of pituitary microadenomas has been reported as hypodense, isodense, and hyperdense. In this study, CT showed a region of diminished enhancement and usually an enlarged pituitary gland in cases of prolactin secreting adenomas. HGH- or ACTH-secreting adenomas were less consistently hypodense. It is concluded that hypodensity and enlargement in the pituitary gland are the most useful criteria for identification of microadenomas. Some technical factors that may affect the CT appearance of microadenomas and lead to conflicting reports are discussed. PMID- 6294735 TI - Acute esophageal food impaction treated by gas-forming agents. AB - Impacted meat obstructing the esophagus was successfully treated in 8 patients by oral administration of tartaric acid and sodium bicarbonate. This acid-base mixture produces carbon dioxide, which distends the esophagus and propels the meat into the stomach. The authors have had a 100% success rate with no complications and recommend that this procedure be the first treatment attempted for meat impaction in the esophagus. PMID- 6294736 TI - Multilocular cystic nephroma: a radiographic-pathologic correlation of 58 patients. AB - Clinical radiographic, and pathologic findings of multilocular cystic nephroma (MLCN) in 58 patients are presented. The lesion that affects predominantly boys in childhood and women in adulthood is usually solitary but rarely can be multiple, and it commonly occurs as an asymptomatic mass, occasionally with hematuria. Tumors may grow slowly over years or rapidly within months. A mass that is usually identified on plain radiographs occasionally has curvilinear calcification. Excretory urography and retrograde pyelography are helpful when pelvic herniation of the tumor is recognized, or when septae are noted with total body opacification. Angiography showed an avascular mass (six lesions), a hypovascular mass (14 lesions), or a hypervascular mass three lesions). Ultrasonography is helpful when multiple circumscribed sonolucent areas are identified. Similarly, a multilocular mass with septae is usually identified by computed tomography (six of seven cases). Our findings and a review of the literature indicate that this lesion is a neoplasm that can be strongly suspected preoperatively if pelvic herniation or a multilocular pattern is noted on imaging studies. PMID- 6294737 TI - Ultrasound, x-ray mammography, and histopathology of cystosarcoma phylloides. AB - Ultrasound and x-ray mammograms were obtained in 8 patients with palpable breast masses, which were subsequently proven to be cystosarcoma phylloides. Histopathologic examination of the 8 tumors showed benign cystosarcoma phylloides in 5, recurrent benign tumor in 1, coexistent invasive papillary carcinoma in 1, and coexistent in situ duct cancer and in situ lobular cancer in 1. A retrospective review of the B-scan images was undertaken to list the descriptive ultrasound features of cystosarcoma phylloides. The ultrasound findings included low-level internal echoes, smooth walls, and good through transmission. The carcinomas were indistinguishable from the benign lesions, although ultrasound was able to distinguish the cystosarcoma phylloides lesions as predominantly solid lobulated masses. PMID- 6294738 TI - Comparative value of bone scintigraphy and radiography in monitoring tumor response in systemically treated prostatic carcinoma. AB - Radionuclide bone scans and skeletal radiographs were obtained before and during combination chemotherapy or initial hormonal treatment in 46 patients with disseminated adenocarcinoma of the prostate. The purpose of the study was to determine the usefulness of these two modalities in evaluating tumor response to therapy. Prior to treatment, bone scans were positive in 44 patients (96%). In all but one patient either bone radiographs or bone marrow biopsy revealed evidence of osseous metastases. In 22 patients partial response to therapy was documented by a variety of other staging tests. Eleven of these patients showed concurrent or later improvement on bone scans; one showed improvement on a radiograph. "Flare phenomena" were observed relatively frequently since 23% of the scans and 50% of the radiographs showed worsening at the time of response. Bone scans revealed worsening in 79% of 33 patients with disease progression of extraosseous tumor; radiographs were equally sensitive (82% worsening). It is concluded that bone scans in particular are useful for monitoring tumor status in systemically treated patients with prostate cancer. However, because of the lack of sensitivity for response and paradoxical worsening with tumor regression in some patients, scans are not accurate enough to be employed as the sole test in following these patients. PMID- 6294739 TI - Nonspecificity of the "rim sign" in the scintigraphic diagnosis of missed testicular torsion. AB - Causes of a hyperemic peritesticular rim on dynamic and static scintigrams are reviewed. Of 6 patients exhibiting such a pattern, 3 had missed testicular torsion; the other 3 had tumor, trauma, or inflammation. The authors conclude that a hyperemic peritesticular rim is a nonspecific finding reflecting underlying pathophysiological changes and is not pathognomonic of missed torsion. PMID- 6294740 TI - Treatment of congestive heart failure. II. Vasodilators and angiotensin converting enzyme inhibitors. PMID- 6294742 TI - Calcium and histamine secretion from mast cells. PMID- 6294743 TI - [Hormone receptor and stimulated turnover of inositol phospholipids; role of PI response in the enhancement of Ca2+ influx]. PMID- 6294741 TI - Physiology of calcium current in cardiac muscle. PMID- 6294744 TI - [Pathology of mitochondria with special reference to megamitochondrial formation]. PMID- 6294745 TI - [Somatic cell genetics on cultured mammalian cells--drug-resistant and auxotrophic mutants]. PMID- 6294747 TI - Effects of prostaglandins, dibutyryl cAMP, LHRH, estrogens, progesterone, and potassium on output of prostaglandin F2 alpha, 13, 14-dihydro-15-keto prostaglandin F2 alpha, hCG, estradiol, and progesterone by placental minces. AB - In order to compare the endocrine response of placental minces to luteinizing hormone releasing hormone (LHRH) and dibutyryl cAMP (dbcAMP) and to screen for effects of potential stimulatory and inhibitory substances, the simultaneous outputs of PGF2 alpha, 13, 14-dihydro-15-keto-prostaglandin F2 alpha (PGFM), progesterone, 17 beta-estradiol, and hCG were evaluated during a 4 hour incubation in 5 placentas. The output of hCG was highest for 12-week placentas, intermediate for a 16 week placenta, and lowest for term placentas. The output of 17 beta-estradiol by 12 and 16 week placentas in the presence of 30 microM dehydroepian-drosterone sulfate (DHEAS) was greater than that by term placentas. Progesterone output was apparently independent of gestational age although some variation between 12-week placentas was demonstrated. Output of PGF2 alpha was lower in 12 and 16-week placentas than in term placentas and that of PGFM was lower in 12-week placentas than in term placentas. LHRH (100 nM) produced stimulation of PGF2 alpha output (P less than .005) and a trend toward inhibition of progesterone output (which failed to achieve statistical significance) but no stimulation of hCG under these conditions. Stimulation of the outputs of hCG (P less than .005) and PGF2 alpha (P less than .001) and inhibition of that of progesterone (P less than .005) was produced by 20 mM dbcAMP. DHEAS inhibited output of progesterone (P less than .01) and PGF2 alpha (P less than .01). There were no effects of potassium, estrogens, progesterone, or prostaglandins on output of any measured substance. PMID- 6294746 TI - Prostaglandins and the rat seminal vesicle: effects of mating and adrenergic stimulation. AB - The concentrations of PGE, PGF, and 6-keto-PGF1 alpha were increased in rat seminal vesicle tissue following mating activity. Likewise, synthesis of PGE and PGF was stimulated by epinephrine (3 X 10(-7) to 3 X 10(-6) M) in tissues and media from in vitro incubations of intact rat seminal vesicles. The in vitro stimulation was inhibited by phentolamine, an alpha-adrenoreceptor blocking agent. Carbamylcholine (2 X 10(-6) M) and bradykinin (1 X 10(-6) M) had no effect on PGE or PGF synthesis, even though both compounds stimulated contractility of the rat seminal vesicle at these concentrations. These data suggest that mating and adrenergic stimulation increase prostaglandin synthesis in the rat seminal vesicle, probably through an alpha-adrenergically mediated mechanism. PMID- 6294748 TI - Short-term flavour memory in the rat. PMID- 6294749 TI - Medical surveillance of biotechnology workers: report of the CDC/NIOSH Ad Hoc Working Group on Medical Surveillance for Industrial Applications for Biotechnology. PMID- 6294751 TI - Delta versus mu receptors: cardiovascular and respiratory effects of opiate agonists microinjected into nucleus tractus solitarius of cats. AB - The cardiovascular and respiratory responses to relatively specific mu or delta agonists microinjected (0.5 microliter/kg) into the region of the nucleus of tractus solitarius (NTS) were examined in anesthetized cats. Blood pressure, heart rate, and respiratory rate were monitored for 30 min after the microinjection of opioid compounds or saline vehicle. The delta agonist, (D Ala2,D-Leu5)-enkephalin (10-100 nmol/kg) elicited dose-dependent decreases in blood pressure, heart rate, and respiratory rate which were naloxone reversible. In contrast the mu agonists, morphine (10-54 nmol/kg) and morphiceptin (100-320 nmol/kg) had no effect on blood pressure or respiratory rate; yet, naloxone elicited pressor responses in animals pretreated with these mu agonists. A receptor-binding assay demonstrated a predominance of mu sites in the NTS. These data show that the delta opiate agonist is more effective than mu agonists in modifying cardiovascular variables in the NTS; we suggest caution in relating specific cardiovascular function to receptor subtypes defined by binding assays. PMID- 6294750 TI - Effects of PHI on vasoactive intestinal peptide receptors and adenylate cyclase activity in lung membranes. A comparison in man, rat, mouse and guinea pig. AB - The presence of receptors, recognized by vasoactive intestinal peptide (VIP) as well as by PHI (a peptide with N-terminal histidine and C-terminal isoleucine amide), was documented in lung membranes from rat, mouse, guinea pig and man by the ability of these receptors, once occupied, to stimulate adenylate cyclase. In lung membranes from rat, mouse and guinea pig, the capacity of VIP, PHI and secretin to stimulate the enzyme and the potency of the same peptides to compete with 125I-VIP for binding to VIP receptors were similar, the affinity decreasing in the order: VIP greater than PHI greater than secretin. In addition, dose effect curves were compatible with the coexistence of high-affinity and low affinity VIP receptors, in the four animal species considered. If PHI was able to recognize all VIP receptors it could not, however, discriminate the subclasses of VIP receptors. PMID- 6294752 TI - Effect of beta-endorphin on gastric acid secretion and serum gastrin concentration in humans. PMID- 6294753 TI - [Use of the stable isotopes in life science (VII). Analysis of stable isotope tracers by nuclear technics]. PMID- 6294754 TI - [Use of the stable isotopes in life science (VIII). Proton activation analysis of 18O in biological samples]. PMID- 6294755 TI - [After care of patients treated with partial prosthesis]. PMID- 6294756 TI - [Liberation of Schistosoma mansoni, Sambon, 1907 cercariae by Biomphalaria glabrata (Say, 1818) maintained in a medium with magnesium thermophosphate]. PMID- 6294757 TI - [Treatment with 1,25-dihydroxycholecalciferol in children with different types of resistant rickets]. PMID- 6294758 TI - [Experimental pharmacology of pirozadil. Comparative study of the action of the product on induced lipolysis in the epididymal fat of rats]. PMID- 6294759 TI - [Transsphenoidal microsurgical treatment of pituitary adenomas]. PMID- 6294760 TI - [Epidemiologic survey of A and B viruses. Study of 500 sera from persons at varying risks]. PMID- 6294761 TI - [Human pentastomiasis in Abidjan. A report on 29 cases]. AB - The authors report 29 cases of human pentastomida detected radiologically in the medical thoracic service over 10 years and a fatal case occurring in the paediatric clinic. After a parasitological review of the life cycle and localisation of the pentastomida in man, the epidemiology clinical picture and typical radiological appearance of thoracic and abdominal calcification are discussed. They are always detected by chance and it is the case history which establishes the link between the eating of poorly cooked snakes or their handling. Finally a case is mentioned (which is currently unique in the literature) of a massive fatal septicaemia in a five years old child caused by Armillifer grandis, which poses the problem of the diagnostic difficulty in current clinical practice of this infestation. Since 1973 the disorder can be diagnosed immunologically thanks to the Marseilles school, but it is difficult to achieve in practice. It would only allow an estimation of the frequency of this infestation and to define the clinical features possible. PMID- 6294762 TI - Differences in effects of amino-terminal and intact parathyroid hormone on calcium, phosphate, and cAMP excretion by the isolated perfused rat kidney. AB - Urinary cAMP (UcAMP) reflects activation of renal adenylate cyclase by parathyroid hormone (PTH). UcAMP excretion and the phosphaturic, anticalciuretic responses to bovine PTH by a stable isolated perfused rat kidney were compared. Dose-response curves were obtained for two synthetic amino-terminal [1-34] PTH preparations and three highly purified intact [1-84] PTH preparations. With all preparations, anticalciuria occurred at lower concentrations (less than 10(-10) M) than those needed to produce a significant increase in UcAMP excretion or phosphate excretion. Maximal decreases in calcium clearance with [1-34] PTH were independent of concentrations between 10(-10) and 10(-7) M whereas cAMP and phosphate increased sigmoidally at PTH concentrations above 10(-10) M. The two synthetic [1-34] PTH preparations had identical anticalciuretic and phosphaturic dose-response curves despite their significantly different effects on cAMP excretion. Intact [1-84] PTH concentration-response curves were identical to those of [1-34] PTH at concentrations less than 3 x 10(-9) M. At higher concentrations, phosphaturia was greater and anticalciuria blunted compared to [1 34] PTH. These latter changes were associated with natriuresis and kaliuresis, effects not seen with [1-34] PTH. The isolated kidney may be a useful model for further studies of PTH action and also constitutes a bioassay system to evaluate hormone potency. PMID- 6294763 TI - Alteration of benzo(a)pyrene metabolism by acute ethanol or corticosterone. AB - The effect of acute ethanol (3 g/kg and 5 g/kg) or corticosterone (12.5 mg/kg) on benzo(a)pyrene metabolism was measured in microsomes from 3-methylcholanthrene pretreated rats. BAP disappearance was inhibited 34% at 5 g/kg but unaltered at 3 g/kg ethanol. Corticosterone administration inhibited BAP metabolism 53%. In adrenalectomized (ADX) rats, alcohol or CS did not decrease BAP metabolism (compared to untreated ADX rats). However, corticosterone in vitro inhibited BAP metabolism in a dose-related fashion in washed microsomes from untreated rats. These results demonstrate that acute ethanol and corticosterone administration alter benzo(a)pyrene metabolism. Furthermore, the inhibition requires an intact adrenal. PMID- 6294764 TI - Influence of pH on ketoconazole activity against Candida parapsilosis and Candida albicans. AB - Experiments involving viable count determinations during incubation of yeasts in a synthetic liquid medium showed that ketoconazole was significantly more active at pH 7.0 than it was at pH 5.5 against strains of Candida parapsilosis and Candida albicans. PMID- 6294765 TI - Influence of dietary purines on pool size, turnover, and excretion of uric acid during balance conditions. Isotope studies using 15N-uric acid. AB - Pool size, turnover, and excretion of uric acid were investigated in three normal subjects both during purine-free, isoenergetic liquid formula diet and during additional purine administration by use of isotope dilution techniques. The fractional turnover of the uric acid pool was increased during dietary purine administration suggesting an increased total body uric acid clearance as a result of the increase in renal clearance. Fractional turnover increased more in the female subject than in males, while pool size was increased less. It can be calculated from the results obtained that endogenous uric acid synthesis is not inhibited by dietary purines. PMID- 6294766 TI - An electronic system of infrared spectral data for toxicological analysis. PMID- 6294767 TI - Prospects for the use of assisted circulation in cases of poisoning. PMID- 6294768 TI - Hemodynamic studies of the effect of total alkaloids of datura in experimental hemorrhagic shock in dogs. PMID- 6294769 TI - The concentration of ammonia in blood and plasma stored for transfusion. PMID- 6294770 TI - An international collaborative clinical study mechanism for resuscitation research. AB - Recent experimental and clinical studies of cardiac arrest, suggesting a brain damage ameliorating effect of thiopental loading, stimulated the development of an international randomized clinical trial of brain resuscitation. Twelve collaborating hospitals in 9 countries are testing the efficacy of post-cardiac arrest thiopental loading (30 mg/kg body wt). The methodology for assessing insult and outcome data, as well as risk and benefit monitoring, is described. This clinical trial will be completed in 1983 and is expected to provide valuable data about the efficacy of thiopental loading. The study mechanism is now well established and ready for the evaluation of future promising brain resuscitation therapies. PMID- 6294771 TI - Serum phospholipase A2 and pulmonary changes in acute fulminant pancreatitis. AB - Twenty-three patients with acute fulminant pancreatitis were studied. The diagnosis was confirmed at laparotomy in every case. Blood samples for the assay of phospholipase A2 were collected for 14 days, and the pulmonary status of the patients was followed by monitoring the blood gases and the inspired oxygen fraction and studying a derived variable, the alveolar to arterial oxygen tension difference--the arterial oxygen tension ratio (A--aDo2/PaO2). The serum phospholipase A2 activities correlated with the changes in pulmonary function and with the outcome of the disease. Eight patients succumbed and they showed higher phospholipase A2 activities and A--aDo2/PaO2 ratios than the five patients who survived after major complications and the ten patients who survived without major complications. The results suggest that in acute fulminant pancreatitis serum phospholipase A2 activity correlates with the severity of the pulmonary changes. Furthermore, it seems to reflect the prognosis. PMID- 6294772 TI - Cerebral protection during extracorporeal circulation. PMID- 6294774 TI - Optimal inputs for parameter determination of inert gas washout from the lung. AB - The nitrogen washout test will yield more information about the distribution of pulmonary ventilation if a pattern of inspired gas concentrations is utilized other than the standard series of 100% oxygen breaths. The input breathing pattern which yields optimal results will vary with the specific features of the lung being studied but typically includes breaths of air, particularly in the last third of the washout. Using computerized, mathematical techniques, optimal inputs were selected for washout tests of duration 10, 20, 30 and 40 breaths for each of six lung models, ranging from a unicompartmental to a highly non-uniform 'diseased' lung. Knowing these optimal inputs we were able to devise a nominal input which was similar to the optimal inputs for all models. For a 10 breath test this nominal input pattern utilized a breath of air at breath 7. For a 20 breath test, air is utilized at breaths 12, 14, 15, 19; for 30, air at 19, 21, 22, 23, 29 and for a 40-breath test, air at breaths 19, 24, 28, 30, 31, 32 and 39. The 30-breath nominal sequence was compared with the conventional oxygen-only washout and a 31-breath pseudo-random binary (PRB) input. Comparisons were made using computer simulated washouts as well as actual tests on human subjects. The 30-breath nominal input was always superior to the standard washout and was superior to the PRB input for all models except the lung ventilated with vital capacity breaths. We conclude that a significant advantage is gained by substituting the recommended sequences of breaths in studies of the dynamics of gas exchange in the lung. PMID- 6294773 TI - Diagnostic vitrectomy in metastatic breast carcinoma in the vitreous. AB - Pars plana vitrectomy was employed to establish the diagnosis of intraocular carcinoma in a 72-year-old woman with a history of breast carcinoma and cerebral astrocytoma. Cytologic study of the material obtained from the vitreous cavity established the diagnosis of metastatic breast carcinoma, and this was later confirmed by study of the eyes obtained postmortem. The clinical and cytologic features of the ocular metastasis of breast carcinoma are presented. PMID- 6294775 TI - [Advantage of the serum measurement of angiotensin I converting enzyme in sarcoidosis]. PMID- 6294776 TI - [Treatment of superficial mycoses with ketoconazole]. PMID- 6294777 TI - Current status of cefotaxime sodium: a new cephalosporin. A symposium. Phoenix, Arizona, January 12 and 13, 1981. PMID- 6294778 TI - Mechanisms of bacterial resistance to antimicrobial agents, with particular reference to cefotaxime and other beta-lactam compounds. AB - Many mechanisms exist by which bacteria can become resistant to antimicrobial agents. Although mutational events are important in the development of resistance to some agents, by far the most important factor in resistance is extrachromosomal genetic material in the form of plasmids. Important mechanisms of bacterial resistance to antibiotics are interference with the transport of the antimicrobial agents into the bacterial cell, inactivation of the agent, and alteration of the target site or metabolic pathway by the microorganism. In order to affect bacteria, beta-lactam antibiotics must pass through proteins at the surface of the cell, must evade destruction by beta-lactamases, and must have an affinity for proteins involved in cell wall synthesis. A number of the new beta lactam antibiotics, because of their great resistance to destruction by beta lactamases and their high affinity for penicillin-binding proteins, have overcome many of the mechanisms bacteria have used in the past to resist beta-lactam drugs. PMID- 6294779 TI - Cefotaxime: a review of in vitro antimicrobial properties and spectrum of activity. AB - Cefotaxime has remarkable potency against all Enterobacteriaceae, including Enterobacter species, Citrobacter freundii, Serratia marcescens, and Morganella morganii, Proteus vulgaris, and Providencia species--all of which are resistant to earlier cephalosporins. Cefotaxime generally inhibits greater than 90% of enteric bacilli at concentrations of less than or equal to 0.5 microgram/ml; in one study it inhibited greater than 98% of isolates at less than or equal to 8 micrograms/ml. For staphylococci and nonenterococcal streptococci, the mean values for the minimal inhibitory concentration50 (MIC50) of cefotaxime (i.e., the lowest concentration inhibiting growth of 50% of tested strains) are 1.1-1.9 microgram/ml and 0.01-0.05 microgram/ml, respectively. Cefotaxime is inactive against Streptococcus faecalis and most other serogroup D streptococci. It is moderately active against Pseudomonas aeruginosa (MIC50, 19 microgram/ml) and Acinetobacter calcoaceticus subspecies anitratus (MIC50, 18 microgram/ml). Because the activity of cefotaxime against other pseudomonads and nonfermentative gram-negative bacilli varies, in vitro susceptibility testing must be used as a guide to therapy. Cefotaxime is potent against Haemophilus influenzae and Neisseria species. Infections due to beta-lactamase-producing gonococci have been treated effectively with cefotaxime (MIC, mode = less than or equal to 0.004 microgram/ml). Most anaerobes are highly susceptible to cefotaxime, but the minimal inhibitory concentrations for 10%-20% of Bacteroides fragilis strains (MIC50, 5.3 microgram/ml) and other Bacteroides species may exceed obtainable serum concentrations. The potent antimicrobial activity of cefotaxime appears to be the result of a combination of characteristics which include: beta-lactamase stability (types I, III, IV, and V), good ability to pass through the cell membrane, strong affinity for lethal penicillin-binding proteins 1a, 1b(s), and 3, minimal limitation by the inoculum effect, and bactericidal action at or close to the inhibitory concentration. Clinically useful methods of susceptibility testing have been developed and can be recommended for clinical laboratory use. PMID- 6294780 TI - Antimicrobial susceptibility tests with cefotaxime and correlation with clinical bacteriologic response. AB - Cefotaxime is a new cephalosporin with a wide spectrum of antimicrobial activity. For broth and agar dilution tests, the MIC breakpoints suggested for defining bacterial susceptibility are less than or equal to 8 microgram/ml for susceptible, 16-32 microgram/ml for moderately susceptible (indeterminate), and greater than 32 microgram/ml for resistant. For disk diffusion tests, the zone diameter breakpoints suggested are greater than or equal to 23 mm for susceptible, 15-22 mm for moderately susceptible, and less than or equal to 14 mm for resistant. Examination of the bacteriologic outcome of infection for 1,440 patients treated with cefotaxime shows that the results of disk diffusion tests correlated well with clinical response. The organisms that had cefotaxime disk zones of greater than or equal to 23 mm were eradicated in 89% of cases. Clinical response of urinary tract infections due to Pseudomonas aeruginosa was unrelated to disk results for bacterial isolates; the rate of bacteriologic cure was 55% 64% regardless of category. It is further recommended that cefotaxime and other cephalosporins not be tested against the enterococci. PMID- 6294781 TI - Metabolism of cefotaxime in animals and humans. AB - The metabolism of [14C]cefotaxime was studied in vivo in rats, dogs, and humans and in vitro in cells of rats and rabbits. Excretion of radioactivity was similar in all species, and greater than 80% of the dose was recovered in the urine. Approximately one-third of the dose was eliminated unchanged, and the major metabolite was desacetylcefotaxime. Under normal circumstances these two products, both with antibacterial activity, were the only materials detected in the plasma. Two further metabolites, designated M2 and M3, (formerly known as UP1 and UP2, respectively, were observed in canine and human urine. Although M2 and M3 were not present in the plasma of normal animals, they were found in the plasma and bile of nephrectomized rats. Extensive studies have shown that the metabolic pathway follows the route: cefotaxime leads to desacetylcefotaxime leads to desacetylcefotaxime lactone leads to M metabolites. The rate-limiting step is the formation of desacetylcefotaxime lactone. All of these reactions take place in the liver. It is concluded that species differences in the metabolism of cefotaxime are more likely to be quantitative than qualitative and that both rat and dog are suitable species for toxicity studies. PMID- 6294782 TI - Clinical pharmacokinetics of cefotaxime in patients with normal and reduced renal function. AB - Two analytic methods for measuring cefotaxime, microbiologic and high-pressure liquid chromatography (HPLC), were compared in normal healthy subjects and in patients with impaired renal function. Results of studies showed that in healthy subjects the levels of desacetylcefotaxime are low relative to those of cefotaxime and that either a bioassay or HPLC is adequately specific for determination of pharmacokinetic parameters. However, in patients with decreased renal function, desacetylcefotaxime reaches much higher levels than in healthy subjects. Thus, the less specific bioassay produces misleading data on serum and urine concentrations in these patients, and the HPLC assay of cefotaxime is recommended for estimating pharmacokinetic parameters in patients with decreased renal function. In healthy subjects and in patients with a creatinine clearance greater than 20 ml/min, neither cefotaxime nor its metabolites accumulates after multiple dosing. If creatinine clearance is less than 20 ml/min, metabolites begin to accumulate. It is recommended that until further data are obtained the dose of cefotaxime given to patients with an estimated creatinine clearance of less than 20 ml/min be half that given to patients with values greater than 20 ml/min. PMID- 6294783 TI - Levels of cefotaxime in body fluids and tissues: a review. AB - Cefotaxime is a third-generation cephalosporin with a broad spectrum of activity. Concentrations of cefotaxime in serum and urine are sufficient for clinical efficacy. This report reviews the available data on the penetration of cefotaxime into other body fluids and tissues. Therapeutic doses of cefotaxime result in significant levels in bile (20 micrograms/ml), in cerebrospinal fluid of patients with meningitis (5-10 micrograms/ml), in pleural fluid (2-7 micrograms/ml), and in otitis media exudate (2-10 micrograms/ml). Lower levels of cefotaxime are obtained in uninflamed aqueous humor (1 microgram/ml), in breast milk (0.1-0.5 microgram/ml), and in cerebrospinal fluid of patients without meningitis (0.2 microgram/ml). Cefotaxime levels in tissues are generally well within the range required for clinical efficacy--i.e., 2-5 micrograms/g, with some higher levels reported in testis, prostate, ureter, skin, and gallbladder wall. Drug levels of less than 2 micrograms/g have been reported in fat, muscle, and uterus. All body fluids and tissues, including bone and skin, are penetrated easily by therapeutic doses of cefotaxime. PMID- 6294785 TI - Antimicrobial activity of desacetylcefotaxime alone and in combination with cefotaxime: evidence of synergy. AB - The level of antimicrobial activity of 3-desacetylcefotaxime was found to be approximately eightfold lower than that of cefotaxime. Variation in the inhibitory effects of desacetylcefotaxime was species dependent. Desacetylcefotaxime was 1-2 log2 dilutions more active than cefotaxime against Pseudomonas acidovorans and Pseudomonas cepacia. Desacetylcefotaxime and cefotaxime were bactericidal, and their minimal inhibitory concentrations were influenced only slightly by increases in the inoculum concentration. Synergy (complete and partial) of 75.8% was noted with the combination. Only with strains of Morganella morganii was the combination antagonistic, but these results were not clinically significant; i.e., all strains remained susceptible to cefotaxime. PMID- 6294784 TI - Cefotaxime toxicity studies: a review of preclinical studies and some clinical reports. AB - Toxicologic studies indicate that cefotaxime is well tolerated by animals. It is difficult to demonstrate acute toxicity. Subacute and chronic effects include local reactions at the site of injection, some hematologic changes with enlargement of the spleen, cecal enlargement in rats and mice, and slight changes in kidney tubules. Special studies indicate that the potential for nephrotoxicity in animals is minimal, and limited clinical reports show no nephrotoxicity in humans. In animals cefotaxime is well tolerated when combined with furosemide, gentamicin, lidocaine, or probenecid. Intrathecal or suboccipital injections of cefotaxime produce convulsions. At concentrations studied cefotaxime was not mutagenic, and no effects on fertility or teratology were observed in animals. PMID- 6294786 TI - Antibacterial activity of desacetylcefotaxime alone and in combination with cefotaxime. AB - Cefotaxime is metabolized to 3-desacetylcefotaxime. The antibacterial activity of desacetylcefotaxime is four- to eightfold less than that of cefotaxime. The activity of desacetylcefotaxime is species- and strain-specific. Although desacetylcefotaxime is less active than cefotaxime, it is more active than cefazolin, cefamandole, or cefoxitin. Desacetylcefotaxime does not inhibit many strains of Morganella, most strains of Pseudomonas aeruginosa, some strains of Serratia marcescens, some strains of Providencia, and many strains of Bacteroides fragilis at clinically achievable concentrations. Cefotaxime and desacetylcefotaxime act synergistically against 72% of strains tested, and antagonism between the two compounds is seen only with strains of Morganella and Proteus vulgaris. The presence of the metabolite desacetylcefotaxime increases rather than decreases the activity of cefotaxime. PMID- 6294787 TI - The pharmacokinetics of cefotaxime and its metabolites in subjects with normal and impaired renal function. AB - The pharmacokinetics of single doses (15 mg/kg) of cefotaxime and each of its metabolites were examined in six normal subjects and 24 patients who showed various degrees of renal insufficiency. An additional nine patients with renal failure were entered into a multiple-dose study in which cefotaxime (1 g) was given twice daily for five to seven days. After intravenous administration, the levels of cefotaxime in serum declined in two phases. The half-life for the terminal phase was 1.10 +/- 0.78 hr. Cefotaxime administered intramuscularly was well absorbed from the injection site (91% +/- 19%) and produced maximal levels of 25.5 +/- 3.5 microgram/ml in serum at approximately 0.5 hr after injection. The levels of the desacetyl metabolite in serum were comparatively low, and the half-life of this metabolite was 1.3 hr. Microbiologic measurements of cefotaxime showed that the half-life of the drug was lower in normal subjects (1.71 +/- 0.90 hr) than in patients with severe renal impairment (half-life, 10.20 +/- 3.43 hr). However, this difference was due primarily to desacetylcefotaxime. Specific measurement by high-performance liquid chromatography indicated only a slight increase in the half-life of cefotaxime, whereas the increase in half-life of desacetylcefotaxime was greater. The most pronounced increases in half-life of metabolites occurred when values for creatinine clearance in patients were below about 5 ml/min per 1.73 m2. When cefotaxime was administered repetitively, the ratio of maximal serum levels after the last dose to those after the first dose demonstrated minimal accumulation of intact drug. The average accumulation of desacetylcefotaxime in this group of patients was about twofold, and that for metabolites M2 and M3 was approximately fourfold. This accumulation of metabolites was observed only in those patients with severe renal impairment and was not excessive. Hemodialysis caused a significant reduction in the half-life of both cefotaxime and desacetylcefotaxime. Recommendations are made for the adjustment of dosing schedules in patients with renal failure. PMID- 6294788 TI - Penetration of cefotaxime into bronchial secretions. AB - The penetration into bronchial secretions of cefotaxime, a new, highly active cephalosporin, was studied in 45 patients with respiratory infections. Ten patients received 0.75 g and 25 received 1 g of cefotaxime intramuscularly; 10 patients received a 30-min intravenous infusion of 2 g of cefotaxime. Samples of bronchial secretions were obtained by means of fiber-optic bronchoscopy after a single dose in all patients and after three and seven days of treatment in 30 and 15 patients, respectively. Simultaneous serum samples were collected for determination of the relationship between the levels of drug in bronchial secretions and those in serum. Assays were performed by microbiologic agar diffusion. In 30 cases bacteriologic analysis determined the minimal inhibitory concentrations of cefotaxime for the bacteria isolated from sputum. Mean peaks in bronchial secretions reached 1.5-2.5 microgram/ml (according to the groups) after 3 hr; individual concentrations varied according to the dose, the route of administration, and the duration of the treatment. Ratios between the levels in bronchial secretions and the corresponding levels in serum were approximately 8% 25% after 3 hr, as is usual for other cephalosporins. Cefotaxime reached significant concentrations in bronchial secretions, exceeding the minimal inhibitory concentrations for microorganisms responsible for respiratory infections. PMID- 6294789 TI - Penetration of cefotaxime and moxalactam into cerebrospinal fluid of rabbits with experimentally induced Escherichia coli meningitis. AB - Penetration of the new beta-lactam antibiotics cefotaxime and moxalactam into cerebrospinal fluid (CSF) was studied in rabbits with experimentally produced Escherichia coli meningitis. Cefotaxime reached peak concentrations (mean +/- SEM) of 31.9 +/- 5.4 micrograms/ml in serum and 2.8 +/- 0.3 micrograms/ml in CSF after an infusion of 50 mg/kg per hr for 8 hr. Moxalactam, after an infusion of 12.5 mg/kg per hr iv, produced peaks of 31.0 +/- 13.1 micrograms/ml in serum and 9.7 +/- 1.2 micrograms/ml in CSF. Both drugs reduced the initial concentration of E. coli in the CSF by greater than 1,000-fold. An infusion of 100 mg/kg per hr of cephalothin produced a peak concentration of 76.5 +/- 15.2 micrograms/ml in serum but resulted in a concentration of only 0.17 +/- 0.05 micrograms/ml in CSF and had no bactericidal activity in CSF. Paper chromatography of CSF from cefotaxime treated rabbits showed that 85.3% (+/- 3.1%) of the antibiotic activity was ascribed to desacetylcefotaxime, a metabolite that is less potent than the parent drug. Neither cefotaxime nor moxalactam was taken up in vitro by rabbit choroid plexus tissue, but cephalothin was taken up at a rate of 9.6 +/- 1.1 microgram/g per hr. Perhaps cefotaxime and moxalactam reached higher concentrations in CSF than did cephalothin because they are not removed from the CSF by the exit pump of the choroid plexus. The fact that levels of cefotaxime in CSF are lower than those of moxalactam could be attributed to the presence of desacetylcefotaxime, a metabolite that is less active than cefotaxime. PMID- 6294790 TI - Comparative activity of cefotaxime and selected beta-lactam antibiotics against Haemophilus influenzae and aerobic gram-negative bacilli. AB - The activity of cefotaxime was compared with that of ampicillin, moxalactam, and cefoperozone against 50 isolates of Haemophilus influenzae and with that of ampicillin, carbenicillin, cephalothin, cefoxitin, cefamandole, cefazolin, and several other established and investigational beta-lactam antibiotics against several hundred isolates of gram-negative aerobic enteric bacilli. Minimal inhibitory concentrations of the drugs were determined by the agar plate dilution technique for H. influenzae and by the microtiter broth dilution technique for the other pathogens. Cefotaxime was the most active agent against H. influenzae; it was 20 times more active than ampicillin. It was also the most active agent against Escherichia coli, Klebsiella pneumoniae, nontyphoid Salmonella species, and Yersinia enterocolitica. Cefotaxime was among the most active agents against Enterobacter cloacae, Citrobacter species, Shigella species, Proteus mirabilis, and Acinetobacter calcoaceticus. None of the new cephalosporins or penicillin inhibited greater than 90% of the isolates of Pseudomonas aeruginosa at concentrations of less than or equal to 16 micrograms/ml; these drugs were, however, more active than carbenicillin. PMID- 6294792 TI - Clinical experience with cefotaxime in the treatment of patients with bacteremia. AB - Seventy-three of 129 patients with bacteremia were observed for their responses to cefotaxime therapy. The clinical response to treatment with cefotaxime was satisfactory in 68 (93%) of the 73 patients. The bacteriologic response was satisfactory in 64 (92.7%) of the 69 patients with bacteriologic evidence of bacteremia. One hundred twenty-nine patients were observed for adverse reactions. Four patients had adverse reactions serious enough to warrant discontinuation of therapy, and three patients developed superinfections. PMID- 6294791 TI - Comparative susceptibility of Yersinia enterocolitica, Eikenella corrodens, and penicillin-resistant and penicillin-susceptible Streptococcus pneumoniae to beta lactam and alternative antimicrobial agents. AB - The antimicrobial susceptibilities were determined for 14 strains of Yersinia enterocolitica, 28 strains of Eikenella corrodens, 10 strains of penicillin resistant Streptococcus pneumoniae, and 10 strains of penicillin-susceptible S. pneumoniae. Y. enterocolitica was found to be susceptible to the aminoglycosides tested and to chloramphenicol, moxalactam, cefoperazone, and cefotaxime but resistant to ampicillin and variably susceptible to carbenicillin, cefoxitin, and cefazolin. On a weight basis, cefotaxime and moxalactam were the most active agents against E. corrodens. Most strains of E. corrodens were resistant to the aminoglycosides tested: gentamicin, tobramycin, kanamycin, and amikacin. Penicillin-resistant S. pneumoniae was most susceptible to cefotaxime and moxalactam. Cefotaxime was consistently active against all tested strains of Y. enterocolitica, E. corrodens, and penicillin-resistant S. pneumoniae, three unusual but clinically significant pathogens. PMID- 6294793 TI - Treatment of urinary tract infections with cefotaxime: noncomparative and prospective comparative trials. AB - Three studies evaluated the efficacy of cefotaxime for the treatment of urinary tract infections. An open, multicenter, noncomparative trial included 477 patients who received a usual dose of 2 g of intravenous or intramuscular cefotaxime daily for five to 10 days. The maximal daily dosage in severe cases was 12 g. Pathogens included Escherichia coli and species of Citrobacter, Pseudomonas, Klebsiella, Enterobacter, Serratia, Morganella, Providencia, and Proteus. Of all the causative organisms, 83.6% were eradicated in the 271 patients who could be evaluated. The other two studies were prospective, randomized comparisons of cefotaxime (392 patients) with cefazolin (250 patients) for the treatment of urinary tract infections caused by microorganisms susceptible to both antibiotics. The treatment schedule was the same as that in the first study. Results of these two prospective studies were pooled. Cefotaxime eradicated 90% of all pathogens, and cefazolin eradicated 72%. Cefotaxime was significantly more effective (P less than 0.01) than cefazolin in eradicating E. coli and Proteus mirabilis. A satisfactory clinical response was obtained in 98.1% of patients given cefotaxime and in 87.6% of those given cefazolin (P less than 0.01). PMID- 6294794 TI - Clinical trials of cefotaxime for the treatment of bacterial infections of the lower respiratory tract. AB - The efficacy of cefotaxime sodium for treatment of patients with lower respiratory infections was evaluated by three protocols in multicenter trials. The first trial studied cefotaxime alone; the second and third trials compared cefotaxime with cefazolin in observer-blind and single-blind randomized controlled studies, respectively. A total of 656 patients were entered in the three trials; 527 received cefotaxime. Overall rates of bacteriologic and clinical cure, analyzed by pathogen, for the cefotaxime treated patients were 89.9% and 93.9%, respectively. In the two comparative trials, overall rates of bacteriologic and clinical cure with cefotaxime, by pathogen, were greater than 94% and numerically exceeded those for cefazolin in each instance. The efficacies in the comparative studies, when analyzed by total patient responses, were significantly different only for clinical responses to cefotaxime in the single blind randomized trial (P = 0.03). Favorable cure rates with cefotaxime were obtained in patients with infections due to Streptococcus pneumoniae, Haemophilus influenzae, Streptococcus pyogenes, Staphylococcus aureus, and Escherichia coli, and nearly comparable responses were obtained for infections due to Proteus, Enterobacter, and Klebsiella species. Cefotaxime was clinically effective in some of the infections due to Serratia marcescens and Pseudomonas aeruginosa, but bacteriologic failures occurred in 67% and 61% of cases, respectively. All Serratia strains were highly susceptible in vitro to cefotaxime, but the range of minimal inhibitory concentrations of cefotaxime for Pseudomonas isolates was wide. PMID- 6294795 TI - Clinical experience with cefotaxime in obstetric and gynecologic infections. AB - Infections in the upper genital tract continue to be one of the leading causes of serious morbidity for obstetric and gynecologic patients. The polymicrobial, mixed aerobic and anaerobic isolates recovered from women with such infections demand broad-spectrum antimicrobial activity. In the past, combination therapy has been given in order to attain this coverage. In a multicenter open study, cefotaxime was used for treatment of endomyometritis after cesarean section, pelvic cellulitis after hysterectomy, and acute pelvic inflammatory disease. The drug effected a clinical cure in 93% of 104 women. In a randomized comparative study conducted at one center, cefotaxime cured 97% of 36 cases of post-cesarean section endomyometritis; clindamyclin plus gentamicin cured 94% of 18 cases of the same infection. There was no evidence of significant alteration in hematopoietic, hepatic, or renal function with either regimen. Cefotaxime appears to be a safe, extremely effective antimicrobial drug that is ideally suited for single-agent treatment of serious soft-tissue pelvic infections in obstetric or gynecologic patients. PMID- 6294796 TI - Clinical comparison of cefotaxime with gentamicin plus clindamycin in the treatment of peritonitis and other soft-tissue infections. AB - The efficacy and safety of cefotaxime were compared with the efficacy and safety of gentamicin plus clindamycin in the treatment of peritonitis and soft-tissue infection in 112 patients. Patients received 20 mg of intravenous cefotaxime/kg of body weight every 6 hr or 1 mg of gentamicin/kg every 8 hr plus 5 mg of clindamycin/kg every 6 hr (both intravenously). Therapy was continued for five to 10 days. The overall clinical cure rate was 82%, with no significant difference between cure rates in the two groups. Both antibiotic regimens were effective against aerobic and anaerobic isolates, although Pseudomonas aeruginosa, an occasional isolate of Enterobacter, and some anaerobes were resistant to cefotaxime. All clinical failures involved patients who had septicemia or who had received inadequate surgical treatment. Six (11%) of the patients who received combination therapy developed impaired renal function, as indicated by a rise in serum creatinine of 30%. No reduction in renal function was noted in patients given cefotaxime. The clinical efficacy of cefotaxime was equal to that of gentamicin plus clindamycin, and less nephrotoxicity was encountered with cefotaxime. PMID- 6294797 TI - Cefotaxime in the treatment of infections of the skin and skin structure. AB - In a series of open, noncomparative studies, cefotaxime was given to 360 hospitalized patients with bacterial wound infections such as cellulitis, abscesses, or necrotizing ulcers of the skin or subcutaneous tissues. The drug was administered intramuscularly or intravenously in a mean dosage of 4 g per day (range, 1.4-12.0 g per day) in three or four equal doses for at least five days. Clinical response to therapy could not be evaluated for 100 patients, and bacterologic response could not be evaluated for 145 patients. Clinical response was satisfactory in 93.5% of the 260 patients for whom therapy could be evaluated, and bacterial response was satisfactory in 84% of the 225 patients for whom therapy could be evaluated. These rates of response include both single- and multiple-pathogen infections. There were nine instances of superinfection. PMID- 6294799 TI - Treatment of gram-negative bacillary meningitis: role of the new cephalosporin antibiotics. AB - Results of the treatment of gram-negative bacillary meningitis have been disappointing: mortality is extremely high, and treatment with chloramphenicol has shown a high failure rate. This failure rate for chloramphenicol is consistent with the wide gap between minimal inhibitory concentration and minimal bactericidal concentration of this drug for Escherichia coli, Klebsiella, and other Enterobacteriaceae. Cefotaxime, a new cephalosporin, is cidal for most gram negative bacteria at concentrations of less than 0.25 microgram/ml. By late 1981, 137 patients with meningitis due to a variety of bacteria had been treated with this agent. Bacteriologic cure rates were 93% for meningitis due to Streptococcus pneumoniae, Neisseria meningitidis, and Haemophilus influenzae and 88% for meningitis due to gram-negative bacteria (94.4% for meningitis due to E. coli and Klebsiella). This new antibiotic shows considerable promise in the treatment of these forms of meningitis. PMID- 6294798 TI - Treatment of uncomplicated gonorrhea with cefotaxime. AB - Three hundred seventy-six patients with uncomplicated infection due to beta lactamase-negative Neisseria gonorrhoeae were treated with 1.0 g of cefotaxime intramuscularly or with 4.8 x 10(6) units of aqueous procaine penicillin G (APPG) intramuscularly plus 1.0 g of probenecid, administered orally. Cefotaxime eradicated 157 (98.1%) of 160 urethral or endocervical gonococcal infections, 22 (96%) of 23 rectal infections, and 8 (73%) of 11 pharyngeal infections. In comparison, APPG-probenecid eradicated 125 (98.4%) of 127 urethral or endocervical gonococcal infections, 17 (94%) of 18 rectal infections, and 4 (100%) of 4 pharyngeal infections. Of 304 domestic beta-lactamase-negative gonococcal isolates, 294 (96.7%) were inhibited by less than or equal to 0.03 microgram/ml of cefotaxime. In a separate study, cefotaxime eradicated 31 of 31 urethral infections due to beta-lactamase-positive strains of N. gonorrhoeae acquired by U.S. Navy personnel in the Philippines. Treatment with cefotaxime was tolerated better than treatment with APPG, primarily because only one injection is required for cefotaxime. The efficacy of cefotaxime was comparable to that of APPG-probenecid in the treatment of uncomplicated genital or rectal infection due to beta-lactamase-negative N. gonorrhoeae, and cefotaxime appears to be highly effective for the treatment of urethral infection due to beta-lactamase-positive N. gonorrhoeae. Further studies are needed for assessment of the efficacy of cefotaxime for treatment of pharyngeal gonococcal infection. PMID- 6294800 TI - Clinical experience with cefotaxime in the treatment of serious bone and joint infections. AB - Cefotaxime, a new parenteral cephalosporin that is beta-lactamase resistant, was evaluated for safety and efficacy in 55 patients (at 22 hospitals) with serious bone and joint infections. Septic arthritis and bursitis and acute and chronic osteomyelitis were treated with 2-16 g of parenteral cefotaxime per day (mean, 7.45 g) for 4-54 days (mean, 22.8 days). Thirty-seven patients had underlying diseases or conditions, 13 patients had infections that were hospital acquired, and 39 patients required surgery. Staphylococcus was the most frequently isolated pathogen. Overall, 39 of the 51 patients who met all criteria for evaluation had satisfactory responses to cefotaxime. The drug was well tolerated by all patients. Further investigation of cefotaxime for the treatment of bone and joint infections is warranted. PMID- 6294801 TI - Cefotaxime therapy for patients with osteomyelitis and septic arthritis. AB - Cefotaxime, a new cephalosporin, was evaluated for efficacy and safety in the treatment of 52 patients with serious bone and joint infections. For five of these patients therapy could not be evaluated. Diagnosis of osteomyelitis or septic arthritis was made on the basis of clinical and roentgenographic evidence of infection. The diagnosis of a bone infection was confirmed by either a positive culture of a bone biopsy or of blood in combination with a positive bone scan or roentgenogram. The diagnosis of a joint infection was confirmed by a positive culture of joint aspirate samples. Osteomyelitis was arrested in 93% (15 of 16 patients) of cases of acute osteomyelitis, 89% (24 of 27 patients) of cases of chronic osteomyelitis, and 100% (4 of 4 patients) of cases of septic arthritis. Follow-up ranged from 0-17 months after completion of cefotaxime therapy. Laboratory monitoring revealed positive direct Coombs' test (six patients), neutropenia less than 1,000 polymorphonuclear leukocytes (two patients), macular rash (two patients), phlebitis (two patients), and pseudomembranous colitis (one patient). It is concluded that cefotaxime is a useful and safe antibiotic for the treatment of osteomyelitis and septic arthritis. PMID- 6294802 TI - Cefotaxime and cephalosporins: adverse reactions in perspective. AB - Cefotaxime was used to treat infections in 2,579 patients during phase II and phase III clinical trials. This paper summarizes the adverse reactions reported to Hoechst-Roussel Pharmaceuticals (Somerville, NJ) during the treatment of these infections. Cefotaxime caused adverse reactions that are caused by all other cephalosporins, including pain at the site of injection (31.9%), thrombophlebitis (4.9%), skin rash (1.8%), thrombocytopenia (3.8%), glomerulotubular dysfunction (1.4%), diarrhea (1.2%), and superinfection (1.1%). Compared with cefazolin, cefotaxime caused pain on injection, phlebitis, and diarrhea more commonly (P less than 0.05) but caused superinfection less commonly (P less than 0.04). Since these data were obtained from many different sources by diverse methods, further controlled trials are needed to substantiate these differences. However, the adverse reactions caused by cefotaxime appear to be similar in spectrum and severity to those caused by other cephalosporins. PMID- 6294803 TI - [The value of bacteriological controls in the epidemiological studies of tuberculosis]. AB - The authors make an analysis of bacteriological data concerning patients with tuberculosis from a certain area (the Jassy County) which have been recorded between 1971 and 1980. An analysis is presented of the instantaneous and of the maximal prevalence of tuberculous bacilli carriers in the urban and the rural environments, and the incidence of tuberculosis in children and adults, also in the urban and the rural media. Some other aspects are studied of primary drug resistance, as well as of secondary resistance. The data presented in the paper stress the major role of the bacteriological investigation in the epidemiological follow-up of tuberculosis, as well as the fact that bacteriological indicators are capable to evidence essential aspects of the evolution of tuberculosis endemia in a certain area. PMID- 6294804 TI - [Research on the occurrence of functional anomalies of the peripheral bronchi (small bronchi disease) in workers in a factory with respiratory noxious substances]. AB - A group of 908 subjects has been studied, of which two-thirds were males, classified in 5 categories: unexposed, with light exposure, and severely exposed to inhalation of volatile substances, or various types of dust. A B.M.R.C. file was filled for each of the subjects. This file concerns respiratory symptoms. A forced expirogram was also performed, and FEV 25-75%, FEV 75-85%, and TEM were determined. The two expiratory flows (median and final) are frequently disturbed in subjects without respiratory symptoms, more frequently in smokers with a normal FEV 1. In those exposed to respiratory risk the proportion of alterations increases, but association of smoking with inhalation of noxious chemical substances will determine a modification of FEW 1 and anomalies of the peripheral bronchi occur more frequently in smokers. Between the criteria suggesting the presence of small airways disease, the ratio FEV 25-75% under 60%, with a FEV 1/FVC ratio within normal limits, this being the most sensitive of the criteria, also appears as the most reliable. PMID- 6294805 TI - [Current possibilities of echography in pneumological diagnosis (preliminary note)]. AB - Various types of echographies are presented, and it is concluded that the bi dimensional echography, also called type B or echo-tomography is the most useful for the study of pulmonary affections, and of compact formations in contact with the pulmonary tissues. With the aid of Model E.D.B. Unirad sonograph (Technicare, USA), and of the Scintron 5 computer (Scintag, Berthold, Switzerland) 6 patients with pleurisy of various extension, one patient with fibrothorax, and 2 patients with pulmonary tumours have been examined. Five of the cases are presented. Echotomography has demonstrated its usefulness in measuring the thickness of the thoracic wall, in establishing by means of a non-invasive technique the solid or fluid character of pleural affections, in assessing the structure of pachypleuritis and in detecting changes induced by tumours, or atelectasia in the vicinity of the pleura. However, the method does not provide information on round peripheral tumours which do not come into contact with the pleura. Also this method cannot replace other methods already used in the pneumological practice, but can provide supplementary informations without any risks for the patient. PMID- 6294806 TI - [Preliminary results of the use of a new derivative of rifamycin SV in the treatment of sarcoidosis]. AB - Seven patients with advanced forms of mediastino-pulmonary sarcoidosis (involvement of the pulmonary parenchyma, and restrictive respiratory syndrome), confirmed by histopathologic investigation, have been treated with a new SV rifamycin derivative, 1246-EH, or "Reprimum" without any other association. Reprimun was given in daily doses of 10 mg/kg of body weight for a period of two weeks, then, for another 4-5 weeks, the drugs was given intermittently in doses of 15 mg/kg of body weight twice weekly. Between two cycles of therapy a period of 14 days was free of drug administration. In all patients the treatment with the new derivative was well tolerated. The treatment was not followed by adrenal deficiencies, neither was noted obesity, bone decalcification, gastric ulcers, etc., which are commonly associated with prolonged corticoid therapy. The treatment with "Reprimun" efficient for all patients. A clear clinical and X-ray improvement was noted after the first three months of treatment with the new drug. Respiratory function, as well as clinical and radiological signs improved, the serum immunologic parameters, and the biochemical ones were also significantly improved (Ig, C3 total serum proteins and gammaglobulins). All parameters which were tested were finally improved and became normal. In only two of the seven patients the radiological aspects of the lung suggested the presence of sequellae at the end of the treatment. In these two patients there was a persistant reduction in the respiratory volume, as well as a discret hypoxemia. However, both these patients had a more advanced form of the disease, with extensive pulmonary fibrosis, and in these cases prolonged corticoid therapy which had previously been applied had failed. Reprimun had an immunological modulatory effect in all patients, resulting in normal levels of IgG, IgA, and C3 in the first three months of therapy. There was a direct correlation between the normalization of immunological indicators and the good evolution of the diseases, a fact which suggests once again the existence of some immune mechanisms involved in the maintenance of sarcoidosis. PMID- 6294807 TI - [The contribution of the clinical examination to the diagnosis of bronchopulmonary cancer]. AB - An analysis is made of the contribution of clinical examination to the diagnosis of primary bronchopulmonary cancer in a lot of 355 patients. As it is known bronchopulmonary cancer does not have a characteristic and patognomonic clinical symptomatology. In two thirds of the patients (236/66.4%) the disease' onset was marked by respiratory symptoms. In half of these patients (120/33.8% of the total number of cases) the respiratory syndromes for which the patient had requested medical help could have suggested the diagnosis, and were marked by a pseudo pneumonia, or pneumopathy with long evolution, intensified coughing in a patient who had been coughing for a long time, or an onset marked by hemopthysis. In the other half of the patients in this group the onset symptoms had nothing characteristic. The onset marked by metastases was noted 69 of the patients (19.4%), and para-neoplastic syndromes were noted in 10 (2.8%). These certainly suggested the presence of pulmonary cancer. It appears that in over half of the patients 9195, or 54.1%) the clinical examination could suggest the existence of bronchopulmonary cancer, and this percentage increases if patients are included in those whose respiratory symptoms are not characteristic but in those which physical examination will evidence suggestive changes. Only in a very small number of patients (12, or 3.3%) the clinical examination did not provide any diagnostic element. These were the asymptomatic cases, accidentally identified in the course of a radiological examination. It should be stressed that the clinical suspicion of pulmonary cancer should always be confirmed by paraclinical investigations. PMID- 6294808 TI - [Bacteriological examination of the sputum in support of a rational antibiotic therapy of bronchopulmonary infections]. AB - An algorhythm of bacteriological investigation of the sputum is suggested for the etiological diagnosis of acute bronchopulmonary infections with pathogenically conditioned bacteria. This investigation avoids misleading results, and provides, in a minimal time interval, both the first empiric criteria for the application of antibiotherapy--a definition of the microscopy category of the infecting pathogenically-conditioned bacteria--, and the second one, namely the antibiogram of the primary culture. This algorhythm is based on: decontamination of the sample by repeated washings; microscopical screening of the sample's quality, according to the ratio between inflammatory cells, squamous epithelial cells and the fibrin exudate; the qualitative bacterioscopic examination; the semi quantitative culture of the sputum with the antibiogram on the primary culture; identification of infecting pathogenically-conditioned bacteria by confrontation of the microscopic significant amounts; inflammatory cells on the smear prepared directly from the sputum; the antibiogram on sub-cultures of the infecting, pathogenically-conditioned bacteria. Of high significance for the etiologic implication is the association of the inflammatory cells in quantities equal or greater than 13 organisms per microscopic field of pneumoncocoid bacteria, equal or greater than 20 organisms per microscopic field of the haemophilloid germs (with a significance threshold of 5%), and equal or greater than 18 organisms per microscopic field of neisseroid germs (at a significance threshold of 0.27%). PMID- 6294809 TI - [Aspects of the etiological diagnosis of extrapulmonary tuberculous infections]. AB - An analysis is made of criteria used for making an etiologic diagnosis of extra pulmonary tuberculosis over a period of 15 years, and a general correlation is noted between the figures for morbidity and those of bacteriologic confirmations. In the group of patients which has been investigated the diagnosis could be ascertained from urine samples in 44% of the cases, from lymph-nodes and nodules in 15% from pleural fluids in 14%, from purulent collections in 13%, and from the cerebrospinal fluid in 8% of the cases. Human type mycobacteria was evidenced in 90% of the cases. Bovine type of mycobacteria was found in 5% of the cases, with a higher frequency in the cerebrospinal fluid (14%) and in lymph-nodes (11%). In 79% of the cases the antibiogram has evidenced the sensitivity of the isolated strains. Resistance of the strains was noted in 10% of the cases. The strains isolated from lymph-nodes and nodules were resistant in a higher proportion (11%), and those from the pleural fluid in 15%. PMID- 6294810 TI - [Considerations on the results of initial chemotherapy in pulmonary tuberculosis]. AB - The results of initial chemotherapy applied in 501 patients with secondary pulmonary tuberculosis hospitalized and treated between 1973 and 1978 are analized. At the end of the treatment the following results have been noted: 94.8% bacteriological negativation, and 5.2% of cases which had not been influenced by chemotherapy and had constantly presented Koch bacilli in their sputum. From the clinical and radiological viewpoints a persistence of cavitary lesions was noted in 12.1% of all patients, or in 22.4% of those who presented initially with cavities. In the surveillance period 7.5% of the patients again became positive, and another course of therapy solved the new positivity in 84.2% of those involved. A percentage of 95.2% global favourable results have been noted in the final stage of the therapy, and 4.8% failures. Relapses, and bacteriological failures were noted with a higher frequency in males, and are in direct proportion with the extension of the lesions and the severity of the clinical forms. Eighteen of the patients (3.6%) had a course which led to chronicization of the disease. The favourable results and the reduction to a minimum of the therapeutic failures could be obtained by a correct application of chemotherapy and dispensarization of the patients under control. PMID- 6294811 TI - [Tuberculosis associated with bronchopulmonary cancer in a young man]. PMID- 6294812 TI - [Dynamics of the risk of tuberculosis infection and invalidity in Romania]. AB - Analysis of epidemiological indicators is of particular importance, and relevant for assessing the efficiency of the prevention and fighting programs against tuberculosis. The most relevant epidemiometric indicators are the incidence of the infection and that of the disease itself. The annual risk of infection with tuberculous bacilli is considered as the most valuable indicator of extension or limiting tendencies of endemia, and of the efficiency of anti-tuberculosis programs. The annual incidence of the infection at the age of 14 years established by various methods, and improper conditions determined by mass allergizing of the infantile population by mandatory B.C.G. vaccination has decreased from 3,5% in 1960 to 0.8-1.0% in 1980. The level is still high, as compared with other european countries, but the rate of decrease has accelerated over the last years, a fact which demonstrates the efficiency of fighting measures applied. The incidence of contracting tuberculosis should be dealt with separately with regard to children and adults, since in children it is mainly related to primary infection, while in adults it is related to the presence of minimal lesions with phtysiogenic potential which can be traced to the post primary stage, as well as to immuno-depressing influence of some environmental factors. In this connection the incidence of tuberculosis in children has been reduced in our country in the last 25 years from 297.2% to 10.3%. The risk of developing the disease has fallen from 1/230 in 1950 to 1/9 700 in 1980. The risk of developing the disease has decreased more rapidly in older children (between 5 and 9 years) than in the younger ones (0 to 4 years). The incidence of tuberculosis in the population aged above 15 years has fallen from 352.6% in 1955 to 70.5% in 1980. The risk of developing the disease in adults was of 1/284 in 1956 and 1/1 418 in 1980. The authors also make an analysis of the dynamics of some demographic risk factors, such as sex, age and environment. PMID- 6294813 TI - Action of collagenase and elastase from human polymorphonuclear leukocytes on human articular cartilage. AB - Collagenase from human polymorphonuclear leukocytes (neutrophil collagenase) attacks collagen type II in solution at a rate intermediate to those of type I and III collagens. This enzyme alone is not able to initiate degradation of native human articular cartilage. If the cartilage is first treated with leukocyte elastase, collagenase slowly degrades collagen. Confirming earlier findings by other investigators, elastase has a dual action on cartilage: The enzyme removes proteoglycans, thus demasking collagen fibers and giving collagenase access to them, and solubilizes collagen at a sizable rate. Although neutrophil collagenase cleaves collagen type II in solution at a high rate, the native, cross-linked status of collagen in cartilage makes it a relatively poor substrate for this enzyme. On a weight by weight scale, elastase and collagenase display about the same collagenolytic potential on human articular cartilage. The elastase/collagenase system from human polymorphonuclear leukocytes could represent a cooperative proteolytic complex in the destruction of cartilage in rheumatoid arthritis. PMID- 6294814 TI - Immunoglobulin inclusions in rheumatoid arthritis polymorphonuclear cells: lack of correlation with circulating immune complexes. AB - A discriminating direct immunofluorescent test has been used to identify immunoglobulin inclusions in polymorphonuclear leucocytes (PMNs) isolated from the blood of patients with rheumatoid arthritis. These inclusions are thought to represent phagocytosed immune complexes, since normal PMNs incubated in RA sera known to contain raised levels of immune complexes developed similar immunoglobulin inclusions. Inclusions did not develop in normal PMNs incubated in normal serum. No correlation was found between the percentage of either RA blood PMNs with immunoglobulin inclusions or normal PMNs developing inclusions after incubation in RA sera, and levels of immune complexes in the corresponding sera. Using heat-aggregated IgG as a laboratory model of immune complexes, a simple relationship has been demonstrated between the uptake of IgG aggregates by normal PMNs and the concentrations of IgG aggregates in the test solutions over a concentration range of 12.5-200 micrograms . ml-1. These results indicate that the C1q- PEG test gives no measure of the actual amounts of immune complexes available in serum for phagocytosis. PMID- 6294815 TI - Inflammatory responses to intradermal crystals in healthy volunteers and patients with rheumatic diseases. AB - The inflammatory response to intradermal injections of urate, pyrophosphate and hydroxyapatite crystals in human forearm skin is described. Patients with rheumatoid arthritis responded normally to urate crystals, and patients with osteoarthritis or pyrophosphate arthropathy responded normally to hydroxyapatite and pyrophosphate crystals respectively. These results suggest that variation in host response to crystals cannot explain the different patterns of crystal induced disease seen in man. The model, however, is recommended as a safe, simple ethical and reproducible test of inflammation in human subjects. PMID- 6294816 TI - [Granular cell myoblastoma of the tongue]. PMID- 6294818 TI - NMR imaging techniques and applications: a review. PMID- 6294817 TI - [Chemodectoma in an unusual location]. PMID- 6294819 TI - Interaction of insulin, glucagon, and DBcAMP on bile acid-independent bile production in the rat. AB - In experiments on fasted, pentobarbital-anesthetized rats the effect of insulin (1 U X kg-1, followed by 0.05 U X kg-1 X min-1), glucagon (0.5 micrograms X kg-1 X min-1), and dibutyrylic cyclic AMP (DBcAMP) (0.5 mumol X kg-1 X min-1) on bile flow and composit8ion was examined. Infusion of these substances resulted in maximal increases only in the bile acid-independent bile formation, and insulin appeared to be a more powerful stimulant of bile production than glucagon or DBcAMP. When bile production was first stimulated maximally with glucagon or DBcAMP, supplementary infusion of insulin increased bile production significantly. When administration of glucagon or DBcAMP supplemented maximal infusion of insulin, only DBcAMP resulted in a further increase in bile production. Bile production was, however, increased by supplementary glucagon infusion, when a submaximal dosage of insulin was given. No additive effect of glucagon and DBcAMP on bile secretion was observed. The results suggest that glucagon induces choleresis in rats via liberation of cAMP and that the mechanisms of glucagon choleresis differ at least partly from those involved in insulin choleresis. The results are compatible with an insulin-produced inhibition of the adenylate cyclase and activation of the phosphodiesterase in the liver. In accordance with present knowledge of the biological effects of insulin and glucagon the choleretic response to both hormones may be secondary to stimulation of Na-K-ATPase located to the hepatocellular membrane. PMID- 6294820 TI - [Acquired immunologic deficiency syndrome, opportunistic infections and homosexuality. Presentation of 3 cases studied in Switzerland]. AB - Three cases of multiple opportunistic infections in previously healthy homosexual males were observed in 3 different University Hospitals in Switzerland. Two of the patients died. Infections were multiple, with P. carinii pneumonia (3 cases), chronic mucocutaneous ulcers probably due to Herpes simplex virus (2 cases), mucosal candidiasis (2 cases) and disseminated infections due to cytomegalovirus (1 case) and Mycobacterium avium (1 case). All patients had depressed cellular immunity with marked lymphopenia. These cases are similar to those recently observed in the United States, but two of the patients had never been to the USA although both had vacationed in Haiti where the syndrome has been described. These patients illustrate that this new syndrome should also be suspected outside the USA when a patient (especially a male homosexual) presents with persistent fever of unknown origin and develops opportunistic infections without obvious underlying disease. PMID- 6294821 TI - [Fabry's disease. Light and electron microscopic cardiac findings 12 years after successful kidney transplantation]. AB - Fabry's disease (angiokeratoma corporis diffusum universale) is a disorder of sphingolipid metabolism affecting predominantly male patients. Renal damage is usually the cause of death in the fourth or fifth decade. Renal transplantation represents a new approach to correction of uremia and perhaps even of the basic congenital metabolic anomaly. In this case the patient, a 57-year-old man, developed renal failure at the age of 45 and received a renal allograft at that age. The transplantation was followed by complete remission of uremia and concomitant symptoms, but later complicated by progressive cardiac failure, anginal chest pain and arrythmia. Necropsy findings showed persistent deposition of ceramide in the myocardium and especially in the conduction system. This was documented, for the first time to the author's knowledge, by electron microscopic investigations. Cardiac manifestations of Fabry's disease are discussed in relation to patients with renal transplants. PMID- 6294823 TI - [Necrotic enteritis in piglets due to Clostridium perfringens type "C"]. PMID- 6294822 TI - [Hormonal regulation of phosphorus and calcium metabolism at the kidney level]. AB - The effects of hormones in the renal handling of calcium and phosphate are reviewed in the context of transport events occurring in different segments of the nephron. Hormones that influence the renal handling of calcium include parathyroid hormone, calcitonin and vitamin D. However, the kidney plays only a minor role in the maintenance of calcium balance. Hormones that modify the renal handling of phosphate are parathyroid hormone, thyroid hormone, growth hormone, glucocorticoids and vitamin D. In contrast to calcium, the kidney plays an important role in the maintenance of phosphate balance. In this endocrine context the contribution of the kidney is reviewed in clinical conditions of hypo- and hyperphosphatemia. PMID- 6294824 TI - [Regulation of the hypothalamus-pituitary-adrenal cortex axis by locus ceruleus and central noradrenergic system]. PMID- 6294825 TI - [The structural and functional heterogeneity of the axon membrane of mammalian myelinated nerve fibers and saltatory conduction]. PMID- 6294826 TI - [The three-dimensional structure of pharmacology]. PMID- 6294827 TI - Dimethyl sulfoxide stimulates tyrosine residue phosphorylation of rat liver epidermal growth factor receptor. AB - Epidermal growth factor, a potent mitogen, stimulates phosphorylation of its 170,000-dalton plasma membrane receptor. Dimethyl sulfoxide selectively increased phosphorylation of the epidermal growth factor receptor in rat liver microsomal fraction. Maximal stimulation occurred at 15 to 25 percent dimethyl sulfoxide and resembled the effect of epidermal growth factor in magnitude and rapidity. Like epidermal growth factor, dimethyl sulfoxide selectively stimulated tyrosine residue phosphorylation of this protein. PMID- 6294828 TI - Physalaemin: an amphibian tachykinin in human lung small-cell carcinoma. AB - Immunoreactivity to the amphibian peptide physalaemin was characterized from extracts of a human lung small-cell carcinoma by immunological, chemical, and pharmacological means. Tumor-related peptide cross-reacted with three antiserums to physalaemin to yield 1.1 to 1.6 nanomoles per gram of tissue. Physalaemin and tumor peptide had similar retention times on high-performance liquid chromatography after chemical and enzymic modifications that included pH changes, oxone oxidation, use of a hydrophilic ion-pairing reagent, and digestion with trypsin and pyroglutamate aminopeptidase. Both physalaemin and the tumor peptide produced a contractile response of isolated guinea pig ileum at threshold concentrations of approximately 100 to 150 picograms per milliliter. These data suggest that small-cell carcinoma of the lung contains a physalaemin-like peptide that has structural and biological homology to its amphibian counterpart. PMID- 6294829 TI - Recombination during gene transfer into mouse cells can restore the function of deleted genes. AB - Two plasmids containing nonoverlapping deletions of the herpes simplex virus thymidine kinase gene were introduced into thymidine kinase-deficient mouse L cells by DNA-mediated gene transfer. Thymidine kinase-producing transformants were generated by a mixture of the two plasmids at a frequency significantly greater than that generated by either plasmid alone. Southern blot analyses demonstrated that functional thymidine kinase genes were generated by homologous recombination between the two deletion mutants. PMID- 6294830 TI - Protein kinase injection reduces voltage-dependent potassium currents. AB - Intracellular iontophoretic injection of the catalytic subunit of cyclic adenosine monophosphate-dependent protein kinase increased input resistance and decreased a delayed voltage-dependent K+ current of the type B photoreceptor in the nudibranch Hermissenda crassicornis to a greater extent than an early, rapidly inactivating K+ current (IA). This injection also enhanced the long lasting depolarization of type B cells after a light step. These findings suggest the involvement of cyclic adenosine monophosphate-dependent phosphorylation in the differential regulation of photoreceptor K+ currents particularly during illumination. On the other hand, conditioning-induced changes in IA may also be regulated by a different type of phosphorylation (for example, Ca2+-dependent). PMID- 6294831 TI - Evidence for the neuropeptide cholecystokinin as an antagonist of opiate analgesia. AB - The endogenous neuropeptide cholecystokinin, when administered systemically or perispinally, potently antagonizes opiate analgesia produced by foot shock and morphine. Nonopiate foot-shock analgesia is not reduced by this neuropeptide. The spinal cord appears to be a critical site of cholecystokinin action. These experiments suggest a physiological role for cholecystokinin as a specific opiate antagonist in analgesia-mediating systems. A similar mode of action may explain other behavioral effects of cholecystokinin, such as suppression of food intake. PMID- 6294832 TI - Chromosome localization of highly repetitive human DNA's and amplified ribosomal DNA with restriction enzymes. AB - Restriction endonucleases cut and partially removed DNA throughout fixed air dried human metaphase chromosomes. Some enzymes produced a G-banding pattern; some revealed the presence of multiple chromosome-specific classes of highly repetitive DNA in C-band heterochromatin. Enzymes that produced the informative C band patterns had recognition sequences that were four or five, but not six, base pairs long and did not contain a cytosine-guanine doublet. In both rat and human chromosomes, regions containing amplified ribosomal RNA genes were specifically removed by the restriction endonuclease Msp I. PMID- 6294833 TI - A cellular mechanism of classical conditioning in Aplysia: activity-dependent amplification of presynaptic facilitation. AB - A training procedure analogous to differential classical conditioning produces differential facilitation of excitatory postsynaptic potentials (EPSP's) in the neuronal circuit for the siphon withdrawal reflex in Aplysia. Thus, tail shock (the unconditioned stimulus) produces greater facilitation of the monosynaptic EPSP from a siphon sensory neuron to a siphon motor neuron if the shock is preceded by spike activity in the sensory neuron than if the shock and spike activity occur in a specifically unpaired pattern or if the shock occurs alone. Further experiments indicate that this activity-dependent amplification of facilitation is presynaptic in origin and involves a differential increase in spike duration and thus in Ca2+ influx in paired versus unpaired sensory neurons. The results of these cellular experiments are quantitatively similar to the results of behavioral experiments with the same protocol and parameters, suggesting that activity-dependent amplification of presynaptic facilitation may make a significant contribution to classical conditioning of the withdrawal reflex. PMID- 6294835 TI - A benzodiazepine receptor antagonist decreases sleep and reverses the hypnotic actions of flurazepam. AB - The benzodiazepine receptor antagonist 3-hydroxymethyl-beta-carboline, which blocks several of the pharmacological actions of benzodiazepines, induces a dose dependent increase in sleep latency in the rat. Furthermore, at a low dose that by itself does not affect sleep, 3-hydroxymethyl-beta-carboline blocks sleep induction by a large dose of flurazepam. The benzodiazepine receptor may play a role in both the physiological regulation and pharmacological induction of sleep. PMID- 6294834 TI - Associative conditioning of single sensory neurons suggests a cellular mechanism for learning. AB - A cellular analog of associative learning has been demonstrated in individual sensory neurons of the tail withdrawal reflex of Aplysia. Sensory cells activated by intracellular current injection shortly before a sensitizing shock to the animal's tail display significantly more facilitation of their monosynaptic connections to a tail motor neuron than cells trained either with intracellular stimulation unpaired to tail shock or with tail shock alone. This associative effect is acquired rapidly and is expressed as a temporally specific amplification of heterosynaptic facilitation. The results suggest that activity dependent neuromodulation may be a mechanism underlying associative information storage and point to aspects of subcellular processes that might be involved in the formation of neural associations. PMID- 6294836 TI - [Complications of antitumor and antileukemic chemotherapy. 1]. AB - The recent development of chemotherapy in the treatment of cancer and leukemia requires that all practitioners involved have a thorough knowledge of the sometimes life-threatening side-effects of chemotherapeutic agents. All these agents, whether used alone or in a combination, carry a risk because of their lack of specificity which make active on normal cells, especially those with a rapid turn-over such as the hematopoietic cells or the cells of the digestive tract. Prior to the prescription of a chemotherapeutic regimen, the acceptable risk must always be clearly defined, according to the seriousness of the disease and to the patient's age, physical condition and psychological status. During the course continuous monitoring adjusted to the specific toxicity of the agents used is requisite. More or less prominent asthenia and weight loss are common, as the result of various physiopathological mechanisms. Digestive disorders may consist only of nausea and emesis or include mucosal lesions with diarrhea as the main feature. Vincristine and vindesine are responsible for constipation. Hepatic toxicity, which is less common, is usually due to L-asparaginase. Transient hair loss is the most frequent cutaneous side-effect. Hyperpigmentation, photosensitivity, nail lesions, cellulitis and ulcerations may occur, as well as specific lesions with bleomycin. High fever during injection often occurs with this last agent. PMID- 6294837 TI - [Results of the repair of isolated coarctation of the aorta during the 1st 6 months of life, Apropos of 46 cases]. AB - Between 1972 and 1978, forty-six infants under six months of age underwent surgery for isolated coarctation of the aorta. These forty-six patients represent 32% of the total number of infants aged less than six months who had surgery for coarctation of the aorta during the same period. At the time of operation, 41% were aged less than one month and the youngest was five days old. The main cause for surgery was heart failure in infants under one month (14/19) and severe systemic hypertension (150 to 300 mm Hg) in the one to six month age group (18/27). Overall mortality rate in our series was 17%. Six of the eight infants who died were under one month of age. Recoarctation occurred in 31,5% of infants; in eight cases the first surgical procedure had been done before one month of age. A second procedure was necessary in four cases. Early surgical repair of severe coarctation diagnosed during the first six months of life leads to functional improvement and avoids residual hypertension. After repair, the main risk is recoarctation. PMID- 6294838 TI - [Diaphragmatic eventration in adults. Apropos of 20 cases]. AB - Twenty adults with a mean age of 49 were operated on between 1972 and 1980 for eventration of the diaphragm. The etiology was probably traumatic in 11 cases; it was congenital in 2 and degenerative in 7. The functional signs were usually respiratory (55%) or digestive (10%). In all cases, the diagnosis was based on standard fluoroscopy showing superelevation of the hemidiaphragm, which was visualized fluoroscopically as immobile while a pneumoperitoneum showed a absense of any tear. Spirography confirmed a restrictive deficit which was sited broncho spirometrically on the eventration side. Surgical repair was achieved in most cases by thoracotomy (19 cases) and consisted in plication (17 cases) or incision followed by suturing of the overlapping edges of the muscle (3 cases). Postoperative complications included the death of the patient with multiple trauma, on the eight postoperative day, as a result of the cerebrovascular accident, and in another case rutpure of the repair on the second day, requiring a second operation. Apart from one patient who died as a result of a fatal accident two years postoperatively, all the surgically treated cases are alive, and, with one exception, symptom free. After an average follow up period of 2 years and 2 months, the control respiratory function tests show a 21% improvement in vital capacity and a 20% increase in FEV1. On the basis of these results, the authors conclude that this functional surgery is justified in the case of patients presenting with typical clinical symptoms, and worth considering in cases where respiratory function tests show a restrictive deficit, but also in patients who do not complain of breathlessness. PMID- 6294839 TI - [Inguinal island flap for covering trochanteric bedsores]. AB - Trochanteric bedsores are a serious condition because of the danger of osteoarthritis of the coxofemoral joint, which is a source of septicemia, and also because of the difficulty of covering them with skin owing to the mobility of the bone and the absence of skin laxity in the area. The authors suggest a new technique of closure after local preparation providing a clean "surgical" defect. They use a groin island flap raised in the Japanese "retrograde" manner (i.e. laterally to medially). This technique is of course the fruit of the authors' experience of the microsurgery. The distal portion of the flap is drawn, corresponding to the size of the defect (round shape). The proximal portion contains the vessels surrounded by subcutaneous and protected by a de epithelialized triangle of skin with a distal base. The flap is passed through a subcutaneous tunnel between the femoral trigone and the trochanteric area to be covered. The donor site is closed by approximation of the edges after they have been undermined. The authors discuss the others possible surgical managements: 1) local flaps (transposition flap, Z plasties, are unsuitable; 2) the double S rotation flap had the drawback of creating a large undermined area and leaving a scar on the trochanteric region; 3) simple GER-type muscle flaps are not advisable because the transposed muscle always indergoes a fibrous involution so that the graft provides insufficient protection; 4) myocutaneous flaps, and especially fascia lata myocutaneous flap, offer better indication, because of muscle only acts as a vascular pedicle for the skin flap. The only drawback is the relative defect resulting from raising the flap. The authors conclude that the groin island flap offers the following advantages; a skin material which is more than efficient; ideal vascularization; perfect corverage without undue tension of the trochanteric surface; an interesting training for the plastic surgeon performing microsurgical reconstruction. PMID- 6294840 TI - [Ultrasonic diagnosis of acute cholecystitis. Critical study of 50 verified cases]. AB - A retrospective study of fifty surgically proven cases of acute cholecystitis and a review of the literature enabled the authors to summarise the sonographic features of the disease. These may include one or several of the following signs: thick gall bladder wall with an occasional posterior hypoechogenic rim, diffuse echogenicity of the purulent bile and local tenderness at the passage of the transducer. If an abscess is present, there is a hypoechogenic area in contact with the gall bladder. The technical problems of the B scan performed on an emergency basis make the use of real time intercostal scans invaluable. Ultrasound should be the first and may be the radiological examination performed in this situation. PMID- 6294841 TI - [The alcoholic in the post-withdrawal periods. Conditions of their care and results after 2 years. Apropos of 200 cases]. AB - The authors report on two years of their experience in caring for alcoholics after withdrawal. In evaluating results, improvements in asthenia, depression, and anxiety, as well as in the aptitude to resume a normal life, with satisfactory familial, conjugal, sexual, professional and social achievements, are considered. Overall results are given in terms of abstinence, relapse, hospitalization, and normalization of behavior and character. PMID- 6294842 TI - [Chemical sympathectomy in the treatment of arteriopathies of the lower limbs]. AB - Phenol injected transcutaneously into the lumbar sympathetic chain appears to be effective in ischemic arterial disease of the lower limbs. Among the other procedures, such as angioplasty, belonging to the field of therapeutic radiology, this technique seems to have it's specific indications. A preliminary study in fifty patients is reported. PMID- 6294843 TI - [Pathological hyperprolactinemia II. course and diagnostic and therapeutic problems]. AB - Natural growth of prolactinomas unrelated to pregnancy of certain drugs is slow. Mixed adenomas should be systematically looked for since acromegaly or Cushing syndrome may be cured by bromoergocriptine (brc) alone. Management of prolactinomas now rests mainly on bromoergocriptine. Surgery may still be indicated in encapsulated voluminous adenomas, in growing tumors that do not respond to large doses of brc, and in exacerbations during pregnancy which are not rapidly controlled by brc. When hyperprolactinemia does not respond to brc the patient should be investigated for a premenopausal state of a suprasellar lesion. PMID- 6294844 TI - [A therapeutic trial in patients with spasmophilia]. AB - As the clinical manifestations of spasmophilia are characterized by their striking polymorphism, quantitative assessment, which is requisite in order to evaluate therapeutic effectiveness, is a difficult task. The authors have attempted to analyze clinical, electrical and biological data from 30 patients. Each of these patients, given only manganese-cobalt oligosol and magnesium oligosol for two months, in two daily doses, had at least two groups of symptoms. Analysis of results shows that the most significant improvement was clinical: improvement was satisfactory or very satisfactory in 83% of patients tested, with muscular relaxation being recorded first, In relation with these clinical results, is the 59% improvement rate in EMG controls. Improvement in overall laboratory data was conclusive in only 24% of patients; however, if only the changes in intraerythrocytic magnesium, which is the only parameter with real clinical value (60% of abnormal results before treatment) are considered, improvement or return to normal values was recorded in 83% of cases. PMID- 6294846 TI - [Is it necessary to prescribe maintenance therapy in duodenal ulcer?]. PMID- 6294845 TI - [Reflux esophagitis. Etiologies, mechanisms and therapeutic directions]. PMID- 6294848 TI - [Stimulating the immune defenses of the respiratory system]. PMID- 6294847 TI - [What is the status of double contrast barium enema in 1982?]. PMID- 6294849 TI - [Individuality of allergic hypersensitivity to acarids]. PMID- 6294850 TI - [Sleep and respiration in infants and children]. PMID- 6294852 TI - [A questionable terminology for a common disorder: tension headaches]. PMID- 6294851 TI - [Diabetic patients under thirty years of age treated orally]. PMID- 6294853 TI - [Essential tremor]. PMID- 6294854 TI - [Insomnia in daily practice]. PMID- 6294855 TI - [Artificial lenses in surgery for cataracts in adults]. PMID- 6294856 TI - [ORL disorders and air travel]. PMID- 6294857 TI - [Achlorhydric migraine: pathogenesis and management]. PMID- 6294858 TI - [Adverse effects of antitumor and antileukemic chemotherapy. 2]. AB - Aplastic anemia is the most severe hematologic side-effect. All chemotherapeutic agents, with the exception of bleomycin and L-asparaginase, may induce aplasia, but the degree of hematotoxicity varies according to the drug. With the exception of acute leukemia in which drug-induced aplasia is part of the treatment, aplasia must be prevented through perfect knowledge of the posology and injection schedules for each drug, as well as by adjusting doses to the patient's hematological status. If aplasia develops, intensive hematological care is requisite. The most common cardiac side-effect is toxic cardiomyopathy caused by anthracyclines, which must be diagnosed early by EKG recordings before each injection and repeated ultrasonography or dynamic cardiac scintigraphy. The risk of toxic cardiomyopathy makes it requisite not to exceed the maximal doses set for each drug. Pulmonary side-effects include acute hypersensitivity pneumopathy and chronic diffuse interstitial fibrosis, the latter being more common and mainly caused by bleomycin. The risk of chronic fibrosis demands that patients be closely monitored and that the total dose be kept under 300 mg. Renal toxicity usually results in acute transient renal failure, as with cisplatinum, and requires a thorough biological study before each injection. Vesical hemorrhage, which is threatening in some instances, may occur with cyclophosphamide. VM26 and VP16 may induce anaphylactic shock. Allergic symptoms are possible with L asparaginase and bleomycin. PMID- 6294859 TI - [External pelvic endometriosis: diagnostic study. Our experience in the maternity department of Saint-Antoine Hospital. A propos of 130 cases]. AB - The circumstances leading to the diagnosis of pelvic endometriosis (adenomyosis excluded) were studied in 130 patients. From the epidemiological data yielded by these cases, the profile of women susceptible to this disease is delineated. Hysterosalpingography is the key to the diagnosis of endometriosis in most instances; however, coelioscopy and/or laparotomy are required to establish the diagnosis, as well as for staging the disease which is important for the choice of therapy. PMID- 6294860 TI - [Subcutaneous angioma of the hand]. AB - The authors describe the specific features of hemangiomas of the hand in adults (three cases). The clinical syndrome is described. The main examination is by arteriography of the hand which shows three principal aspects: the vascular topography; the volume of the tumors; the extension of the neighbouring tissue. The treatment is analysed: surgical exeresis; in some cases, microvascular transplants are necessary (especially for the distal reconstruction of a finger); embolization is normally avoided because of the risk of necrosis of the finger. PMID- 6294862 TI - [Tiapride and pain in amputees]. AB - Missing limb pain remains a preoccupying and worrisome problem for amputees. Approximately one-third of amputees experience missing limb pain. Although it often abates with time, this pain disables the patient in the first few months following amputation. We carried out a therapeutic trial of oral tablets of tiapride, which was given to twenty patients in three daily doses of 50 to 100 mg each. Tiapride was the sole medication in most instances. Analgesia appeared after an interval of 24 hours to 15 days. Clinical tolerance was excellent. Efficiency in terms of resolution of pain is observed in most cases. PMID- 6294861 TI - [Radioisotopic study of bone growth and maturation. 160 cases]. AB - Increases in blood supply, and certain metabolic and physical processes, are of special significance in epiphyseal growth centers. Such events are evidenced by the uptake of 99mTc methylene diphosphonate (MDP). Bone scans in children and young adults show increased radionuclide uptake in epiphyseal growth plates. Age related changes in 99mTc MDP epiphyseal uptake can be used for determining a functional bone age. Radiation exposure is within the range of other routine diagnostic procedures. Scanning was done with a gamma scintillation camera, which supplies rapid, high resolution, studies. In the first five years of life, nearly all the main epiphyseal growth plates were visible symmetrically on routine scans. In six cases, growth plates were visible before the corresponding epiphyseal ossification foci became apparent on roentgenograms. Little bones (hands, wrists, feet, and ankles) were not assessed because of the poor resolution of the parallel collimator-camera system. A slow decrease in epiphyseal growth plate activity was demonstrated in subjects over sixteen years of age. In some instances, this decrease occurred only after epiphyseal closure became visible on roentgenograms. Data described here are not usually employed for clinical purposes, except when viability of a growth plate is doubtful. Functional analysis with 99mTc MDP is a useful complement to roentgenologic investigation of skeletal growth and maturation. PMID- 6294863 TI - [Eosinophilic fasciitis and Hodgkin's disease]. AB - A case of eosinophilic fasciitis (EF) associated with Hodgkin disease is described. In eight previously published cases, EF was associated with immunologic diseases, such as autoimmune marrow aplasia. The relation between EF and Hodgkin disease should be studied by investigating the physiopathologic meaning of eosinophils in both conditions. PMID- 6294864 TI - [A complication of peritoneojugular shunt for ascites: peritoneal migration of the shunt tubing]. AB - A peritoneojugular shunt operation was done in a cirrhotic patient with ascites. The Le Veen valve functioned correctly for six months. The patient then experienced recurrence of ascites following an uncommon complication: migration of the shunt tubing into the peritoneal cavity. The tubing could no longer be felt in the cervical or thoracic regions. Plain roentgenograms of the abdomen showed that the shunt tubing was in the peritoneal cavity. PMID- 6294866 TI - [Waardenburg's syndrome associated with a generalized epileptic seizure. Cerebral tomodensitometric study]. AB - Waardenburg syndrome was diagnosed in a nine-month-old female infant with dystopia canthorum, broad nasal bridge, overdeveloped eyebrows, cutaneous hypopigmentation, and deafness. Seizures occurred, which makes this case unusual. CAT scan demonstrated wide subarachnoid spaces without ventricular dilatation. PMID- 6294865 TI - [Bilateral ureteral metastases, a 1st symptom of gastric cancer. Gastrectomy. Bilateral ureteral replacement]. AB - Renal colic in a young adult led to the discovery of very extensive bilateral ureteral stenosis. Exploration and biopsy showed metastatic lesions of an asymptomatic gastric cancer. Gastrectomy and subtotal bilateral ureteral replacement by prosthesis gave excellent short term results. PMID- 6294867 TI - [Medium-stay for geriatric patients: a long-stay waiting room or an active rehabilitation unit?]. AB - From May 1979 to April 1980, 223 patients were discharged from a "medium stay" unit (mean length of stay: 45 days) for geriatric patients in Bordeaux (France). The outcome at discharge was established from retrospective data, and the long term outcome (18 months) through a letter sent to general practitioners (rate of response: 88,5% with, in some instances, a prompting phone-call). The mortality rates were 22,5% in hospitalized patients and 38% in survivors after discharge. When those hospitalized for social reasons only are excluded, 62,9% of the remaining patients returned to their former place of residence at discharge. Moreover, 83% were still living at the same place after 18 months (range: 12-24 months). Considering the patients' ages and the seriousness of their conditions, "medium stay", geriatric units seem efficient in helping to maintain the aged at home. PMID- 6294868 TI - [Aging and cerebral vascular sclerosis: myth or reality?]. AB - The most important aspects of normal and pathological brain ageing are considered from a structural, cellular, hemodynamical, chemical and clinical point of view. It seems admitted that cerebral blood flow and cerebral consumption for oxygen decrease with age but when severe health criteria are used to select normal old subjects, no significant differences exist between normal young and optimally healthy elderly. The question arises whether the blood flow reductions are responsible for the alterations in metabolism, or whether it is simply an autoregulatory response to the lesser metabolic demands of the tissue. The part of cerebral arteriosclerosis is questioned on cerebral flow drop accompanying clinical signs in senile mental impairment. Among some cardiovascular risk factors, hypertension seems to play a leading part in acute and chronic neurological diseases associated with age and with certain types of senile dementia (multi-infarct). However in all other cases, hypertension and cerebral vascular insufficiency, for all that this syndrome exists, seem to have been overestimated to explain chronic senile brain deterioration. PMID- 6294869 TI - [Early treatment of myocardial infarction by direct intracoronary thrombolysis]. PMID- 6294870 TI - [How should we treat the acute flare-up of arterial hypertension?]. PMID- 6294871 TI - [Holter recording. Diagnostic and therapeutic applications]. PMID- 6294872 TI - [Should antihypertensive agents be stopped or not before general anesthesia?]. PMID- 6294873 TI - [Management of complete arrhythmia caused by auricular fibrillation]. PMID- 6294874 TI - [Is it necessary to die in mountaineering?]. PMID- 6294875 TI - [Current risks in carotid surgery]. PMID- 6294877 TI - [Tumors of the heart]. PMID- 6294876 TI - [Coronary insufficiency and radioisotopes. Current status of a noninvasive clinical research method]. PMID- 6294878 TI - [At what age should patients with congenital cardiopathies be operated on?]. PMID- 6294879 TI - [Medical treatment of infectious endocarditis]. PMID- 6294880 TI - [When should a patient with an atheromatous lesion of the internal carotid be operated on?]. PMID- 6294881 TI - Radiologic contributions to the diagnosis and management of gynecologic neoplasms. PMID- 6294882 TI - [Ultrastructural study of changes in the rat submandibular gland induced by long term repeated administration of isoproterenol]. PMID- 6294883 TI - Polymicrobial aetiology of genital ulcers. PMID- 6294884 TI - [Cytomegaly in newborn infants. Difficulties in diagnosing clinically manifest and latent forms of infection]. PMID- 6294885 TI - Demonstration of the stem cell nature of clonogenic tumor cells from lung cancer patients. AB - The tumor cell colony-forming efficiency in soft agarose was evaluated in 190 specimens obtained from patients with either small cell carcinoma of the lung (SCCL) or adenocarcinoma of the lung (ACL). Tumor cell colony formation was observed in 45 out of 53 (84%) specimens containing cytopathologically identifiable SCCL tumor cells obtained from a variety of metastatic sites. Tumor cell colony formation was also observed in 6 out of 10 specimens containing ACL tumor cells. The colony-forming efficiency for these lung cancer specimens ranged from 0.003 to 0.72% per nucleated cell plated and from 0.05 to 1.5% per tumor cell plated. There were no differences in plating efficiency for SCCL and ACL. The tumor cell origin of the cells in the agarose colonies was confirmed by cytologic examination, flow cytometric DNA content analysis, and electron microscopy examination. The 'stem cell' nature was demonstrated by the ability of colonies to form typical SCCL or ACL tumors when inoculated into athymic nude mice. In addition, continuous tumor cell lines were established from several specimens forming tumors in nude mice. These data confirm the tumor cell origin and 'stem cell' nature of lung cancer cell colonies growing in soft agarose. Whether the low colony-forming efficiency represents a property inherent in the tumor cells, or a defect in the culture system, remains to be determined. PMID- 6294886 TI - [Lymphoscintigraphy of the retroperitoneum]. PMID- 6294887 TI - Vipoma. PMID- 6294888 TI - The frequency of hepatitis A and B viruses as the offending viral type in suspected hepatitis. AB - Three hundred and thirty blood samples from patients suspected of having hepatitis on clinical grounds but in whom the aetiology of the hepatitis was unknown (93 Whites and 237 Blacks) were tested for the presence of hepatitis B surface antigen (HBsAg), anti-HBs, total anti-hepatitis A virus (HAV) activity and anti-HAV of the IgM class. These tests identified the offending hepatitis virus whenever this was type A or type B, and also revealed the patient's immune status in respect of these viruses. Among the White patients HAV was the cause of 32,2% of the cases of hepatitis and was found commonly in patients up to the age of 35 years, with 1 further example being identified among the patients over the age of 50 years. In contrast, among the Black patients this infection was found only in children, with none of the patients over the age of 5 years remaining susceptible to the disease. Hepatitis B virus (HBV) was found frequently in Blacks of all ages and caused 39,7% of the cases of hepatitis, but was far less common among the Whites, in whom it was responsible for only 10,7% of cases. Serological evidence of exposure to HBV in the combined forms of HBsAg and anti HBs reached 85,7% in Blacks aged between 36 and 50 years, while the highest level to be found in Whites was 45,5%, in the same age group. Active hepatitis A or B infections were diagnosed in 43% of the White patients and 50% of the Black patients in the study. PMID- 6294889 TI - [The cytochemistry of mononuclear-macrophagic cells]. PMID- 6294891 TI - The role of cerebrospinal fluid shunting in tuberculous meningitis. PMID- 6294890 TI - Acute scrotal pathology. PMID- 6294892 TI - [A case of vitamin D resistant rickets]. PMID- 6294893 TI - [Clinical and therapeutic aspects of status asthmaticus in patients with bronchial asthma]. PMID- 6294894 TI - Control of the interferon system: an inhibitor of interferon action. AB - An inhibitor of interferon action has been identified which is present in IFN gamma preparations. The inhibitor is produced following rising interferon concentrations in mitogen stimulated mouse spleen cell cultures. Indications are that the inhibitor is produced in response to the production of interferon, and may therefore be a feedback control mechanism for the interferon system. The action of the inhibitor is independent of both interferon type and concentration, and seems to act by preventing the establishment of the interferon-induced antiviral state at some point following the interaction of the interferon molecule with the cell membrane. The inhibitor has an apparent molecular weight of 8,000 to 10,000 daltons and is stable to treatment with low pH, heat, and trypsin. It is proposed that the inhibitor has the specific role in vivo of controlling interferon-mediated activities. PMID- 6294895 TI - [New ulcer drugs: is cimetidine out of date?]. PMID- 6294896 TI - [Herpesvirus infections in immunosuppressed patients]. PMID- 6294897 TI - [Epidemiologic problems of bacteria resistant to antibiotics: the concept of transposons]. PMID- 6294898 TI - Functional fractionation of platelets: aggregation kinetics and glycoprotein labeling of differing platelet population. AB - Platelet heterogeneity has been studied with a technique called functional fractionation which employs gentle centrifugation to yield subpopulations ("reactive" and "less-reactive" platelets) after exposure to small doses of aggregating agent. Aggregation kinetics of the different platelet populations were investigated by quenched-flow aggregometry. The larger, "reactive" platelets were more sensitive to ADP (Ka = 1.74 microM) than the smaller "less-reactive" platelets (Ka = 4.08 microM). However, their maximal rate of aggregation (Vmax, % of the platelets aggregating per sec) of 23.3 was significantly lower than the "less-reactive" platelets (Vmax = 34.7). The "reactive" platelets had a 2.2 fold higher level of cyclic AMP. Platelet glycoproteins were labeled using the neuraminidase-galactose oxidase--[H3]-NaBH4 technique. When platelets were labeled after reversible aggregation, the "reactive" platelets showed a two-fold decrease in labeling efficiency (versus control platelets). However, examination of whole cells or membrane preparations from reversibly aggregated platelets revealed no significant difference in Coomassie or PAS (Schiff) staining. These results suggest that the large, "reactive" platelets are more sensitive to ADP but are not hyperaggregable in a kinetic sense. Reversible aggregation may cause a re-orientation of membrane glycoproteins that is apparently not characterized by a major loss of glycoprotein material. PMID- 6294899 TI - Further studies on the interaction between thrombin and GP Ib using crossed immunoelectrophoresis. Effect of thrombin inhibitors. AB - The platelet surface protein GP Ib (glycocalicin-related protein) has been shown to be retarded by thrombin-Sepharose 4B in a crossed immunoelectrophoresis system. The interaction between GP Ib and thrombin was abolished when thrombin was blocked either at the active serine site with tosyl-lysine-chloromethyl ketone (TLCK) or phenylmethylsulfonylfluoride (PMSF) or at the fibrinogen binding site (macromolecular binding site) with N-bromosuccinimide (NBS) or heparin, indicating that both sites have to be freely accessible for the retention of the glycocalicin-related protein by thrombin. PMID- 6294900 TI - Inhibitory effect of fibronectin on the fibrin formation. AB - 125I-fibronectin is incorporated into fibrin when blood plasma is clotted with thrombin. This process is dependent upon fibronectin concentration, temperature, and the presence of calcium ions. Fibronectin appears to be a very strong inhibitor of fibrin formation and influences both the thrombin clotting time and fibrin quality. PMID- 6294901 TI - Cleavage of cytoskeletal proteins by two forms of Ca2+ activated neutral proteases in human platelets. AB - Two forms of calcium activated neutral proteases (CANPs) with different affinity to Ca2+ were partially purified from the soluble fraction of human platelets; one (mu-CANP*) required 6 muM Ca2+ and the other (m-CANP*) did 900 muM Ca2+ for the respective half maximal activity. Human platelets were found to contain mu-CANP predominantly in contrast to our previous study in bovine platelets, in which mu- and m-CANP were equally distributed. Among platelet protein preparations examined for possible endogenous substrates of platelet CANPs, actin binding protein (ABP) and 230 K protein were proteolysed completely by mu- or m-CANP in the presence of the respective optimal concentration of Ca2+. As far as mu-CANP is concerned, this is the first demonstration of proteolysis of endogenous substrates, which implies possible involvement of mu-CANP in stimulus-linked platelet reaction coupled with an increase in intracellular Ca2+ to micromolar concentration. Furthermore, microtubules associated proteins (MAPs) of bovine brain was proteolysed by mu- or m-CANP of human platelet in the similar manner, indicating functional ubiquity of CANPS. PMID- 6294903 TI - Some chemical aspects of labeling human fibrinogen with 99m-technetium. AB - Human fibrinogen was labeled with 99m-Technetium. Tin(II)-chloride in citric acid served as reducing agent for the pertechnetate-ion, eluted from a Mo-Tc generator. Before adding the fibrinogen, the citric acid was always neutralized with sodium hydrogen carbonate. The influences on the quantity of fibrinogen bound 99m-Tc of the relative concentrations of Sn(II), fibrinogen and sodium hydrogen carbonate, of the reaction time and temperature were tested by thin layer chromatography. The reaction temperature of 28 degrees C showed an optimum of fibrinogen bound 99m-Tc for the reaction time from 1 and 2 hours. With a reaction time of 30 minutes not enough 99m-Tc was bound to fibrinogen, the doubling of the reaction time from 1 to 2 hours showed only an increase of binding of less than 4%. The concentration of Sn(II) with respect to the fibrinogen concentration showed no influence on the quantity of fibrinogen bound 99m-Tc at low Sn(II)-concentrations. At values higher than 34 times of the fibrinogen concentration a decrease of the quantity of bound 99m-Tc was observed. The concentrations of sodium hydrogen carbonate showed no influence on the quantity of fibrinogen bound 99m-Tc but on the clottability of fibrinogen, the pH of the solution must be approximately 7.5. In 3 parallel and independent experiments under optimized conditions (1 hour at 28 degrees C, molar ratio of Sn(II) : fibrinogen = 8.5, pH = 7.5) 89.97 +/- 0.92 % of 99 m-Tc were bound to fibrinogen. Controls of these results by column chromatography showed a binding of 81.08 +/- 1.47 % of 99m-Tc to fibrinogen. The clottability of fibrinogen tested by the method of Clauss (1) was entirely preserved. PMID- 6294902 TI - The effects of the oral administration of fish oil concentrate on the release and the metabolism of [14C]arachidonic acid and [14C]eicosapentaenoic acid by human platelets. AB - It has been suggested by several investigators that eicosapentaenoic acid (C20:5 omega 3, EPA) might have anti-thrombotic effects. In this experiment, the effect of the oral administration of EPA rich fish oil concentrate on platelet aggregation and the release and the metabolism of [1-14C]arachidonic acid and [(U)-14C]eicosapentaenoic acid by human platelets was studied. Eight healthy male subjects ingested 18 capsules of fish oil concentrate (EPA 1.4 g) per day for 4 weeks. Plasma and platelet concentrations of EPA markedly increased, while those of arachidonic acid (C20:4 omega 6, AA) and docosahexaenoic acid (C22:6 omega 3, DHA) did not change. Platelet aggregation induced by collagen and ADP was reduced. Collagen induced [14C]thromboxane B2 (TXB2) formation from [14C]AA prelabeled platelets decreased. There was no detectable formation of [14C]TXB3 from [14C]EPA prelabeled platelets, and the conversion of exogenous [14C]EPA to [14C]TXB3 was lower than that of [14C]AA to [14C]TXB2. The release of [14C]AA from [14C]AA prelabeled platelets by collagen was significantly decreased. These observations raise the possibility that the release of arachidonic acid from platelet lipids might be affected by the alteration of EPA content in platelets. PMID- 6294904 TI - [A case of syndactylia--'mule foot'--'horse's hoof'--in cattle]. PMID- 6294905 TI - [Rheumatoid diseases in man and animals (1)]. PMID- 6294907 TI - [Fungal rhinitis in the dog. Diagnosis and therapy]. PMID- 6294906 TI - [Infectious bovine rhinotracheitis--infectious pustular vulvovaginitis (IBR/IPV)]. PMID- 6294908 TI - [Adenocarcinoma of the gastric mucosa in the dog]. PMID- 6294909 TI - [Eye diseases in dogs and cats. (8)]. PMID- 6294910 TI - [The contamination of western Europe in 1979 with influenza virus A/equi-2. Vaccination protection of various contingents of horses and vaccination directives for the future]. PMID- 6294911 TI - [Virus and cancer]. PMID- 6294912 TI - [Multiple lung adenomatosis in the dog]. PMID- 6294913 TI - [Hazards of environmental chemicals. Environmental health in the service of human and animal health. 3. Incidence and distribution of environmental chemicals- strategy of environmental health]. PMID- 6294914 TI - Stimulation of 1,25-dihydroxyvitamin D production by parathyroid hormone and dibutyryl 3',5'-cyclic AMP in normal subjects, hypoparathyroidism and pseudohypoparathyroidism. AB - Parathyroid extract (PTE) or synthetic 1-34 human parathyroid hormone (1-34 hPTH) was injected intravenously as a bolus in 4 normal subjects, 4 patients with PTH deficient hypoparathyroidism (HP) and 3 patients with pseudohypoparathyroidism (PHP). In normal subjects and HP, plasma 1,25(OH)2D was markedly increased at 6 hr and reached the peak at 12 or 14 hr after administration of 200 units of PTE or 20 to 30 micrograms of 1-34hPTH. On the other hand, 500 units of PTE or 20 micrograms of 1-34hPTH failed to increase plasma 1,25(OH)2D in PHP. However, 2.5 mg/kg of dibutyryl cAMP remarkably increased plasma 1,25(OH)2D in a patient with PHP. Maximal increments of plasma 1,25(OH)2D in 3 patients with HP(21.7 +/- 5.6 pg/ml, mean +/- S.D.) were nearly as high as in normal subjects (20.6 +/- 7.0 pg/ml), whereas those in 3 patients with PHP (2.3 +/- 2.3 pg/ml) were distinctly lower than in normal subjects or HP. It is suggested that 1,25(OH)2D production by PTH is intact in HP, but is impaired in PHP mainly due to a defect in the activation of adenylate cyclase system. PMID- 6294915 TI - Further observation on empyema after resection of bronchogenic carcinoma and survival. AB - Out of 11,247 cases undergoing resectional therapy for bronchogenic carcinoma at 39 institutions, 6,348 were operated before July 1976 and the remaining 4,899 thereafter. Analyzing both series, the following results were obtained: 1) The rate of pneumonectomies to lobectomies was 1:2.3 in the former and 1:3.9 in the latter. Pneumonectomies were more on the left side, whereas the right lobectomies were about twice as many as the left ones. 2) Postoperative empyema occurred in 180 cases (2.8%) in the former and 79 cases (1.6%) in the latter. The incidence was significantly higher in the cases of right pneumonectomy than in the cases of left pneumonectomy, and in the cases of pneumonectomy than in the cases of lobectomy. Although the incidence of postpneumonectomic empyema decreased only slightly, overall incidence decreased significantly because of marked increase of lobectomy cases. 3) Out of 180 cases of empyema in the former series, 17 died within 30 days. 4) The 5-year survival rate of the patients with empyema was 28 per cent. 5) The 5-year survival rate was 30 per cent in 93 cases of squamous cell carcinoma, 30 per cent in 50 cases of adenocarcinoma and 15 per cent in 13 cases of large cell carcinoma. There was no survivor among 7 cases of small cell carcinoma. 6) Out of 259 cases of empyema, micro-organisms were cultured in 161 cases, were negative in 20, and unknown in 78. Although various organisms were found, gram-negative bacteria and Pseudomonas were most frequently isolated, i.e. 37 per cent and 36 per cent, respectively. 7) As for the treatment of 5-year survival cases of empyema, thoracoplasty and bronchial closure were frequent as compared with open thoracic window technics such as Eloesser's and Symbas' operation. And, more than half of the cases of empyema seem to have been complicated with bronchial fistulae. PMID- 6294916 TI - Effect of aging on lipid and carnitine metabolism. AB - In healthy subjects aged 20 to 50 years, the urinary excretion of carnitine and its serum concentration increased rapidly and markedly after synthetic beta 1-24 ACTH-Z was injected. Their serum triglyceride levels changed inversely. In contrast, healthy subjects older than 70 and patients aged 45 to 50 with atherosclerosis exhibited lower and delayed changes of carnitine excretion and serum concentrations of carnitine and lipid after ACTH injection. When the aged subjects and the atherosclerotic patients were administered with triiodothyronine prior to ACTH, their metabolic responses to ACTH improved towards normal. The results suggest that the aging process impairs the homeostatic regulation of serum lipid through a "lipid-carnitine" system and that thyroid hormone plays a contributory role in the activation of this system. PMID- 6294917 TI - Serological markers of hepatitis A and B virus infection in university students. AB - Serological markers of hepatitis A and B virus infection were determined by radioimmunoassay in 162 university students. Antibody to hepatitis B e antigen (anti-HBe) was demonstrated in 11 of 15 students positive for both antibody to hepatitis B e antigen (HBsAg) and antibody to hepatitis B core antigen (anti HBc). The presence of anti-HBc alone seemed to denote previous hepatitis B virus infection. Antibody to hepatitis A virus was demonstrated only in 3%. Study of 23 stored HBsAg-positive sera showed high titer anti-HBc significantly more frequently in sera positive for anti-HBe than in sera positive for hepatitis B e antigen. Incidence of hepatitis B e antigen was significantly higher in male students with HBsAg, and that of anti-HBe in female students. PMID- 6294918 TI - Deposition, retention, and biological fate of inhaled benzo(a)pyrene adsorbed onto ultrafine particles and as a pure aerosol. PMID- 6294919 TI - Effect of oxygen on the antagonism of cyanide intoxication--cytochrome oxidase, in vivo. PMID- 6294920 TI - The effect of sodium diethyldithiocarbamate treatment on copper and zinc concentrations in rat brain. PMID- 6294922 TI - DDT toxicity: gluconeogenic enzymes and non-specific phosphomonoesterases in three teleosts. AB - In 3 fish species (Labeo rohita, Clarias batrachus and Channa punctatus) subjected to a sublethal concentration (0.035 mg/l) of DDT continuously for 20 days both hepatic and renal acid and alkaline phosphatases were elevated, whereas the rise in glucose-6-phosphatase and fructose-1,6-diphosphatase was restricted to hepatic tissue. The variations in phosphatase levels were more pronounced in L. rohita than in the other two species. PMID- 6294921 TI - Induction of ion-permeable channels by the venom of the fanged bloodworm Glycera dibranchiata. AB - Venom from the poison glands of the polychaete annelid Glycera convoluta has been reported to dramatically increase the frequency of miniature end-plate potentials at the frog and crayfish neuromuscular junctions, without causing detectable ultrastructural changes. We report here that addition of venom from the related annelid Glycera dibranchiata to one side of a lipid bilayer results in the formation of ion-permeable channels in the membrane. The channel forming activity was found in the void volume of a Sephadex G-25 column (mol. wt. greater than 5000). The conductance of a single channel is about 350 pmho in 0.1 M NaCl and is ohmic. The channels exhibit moderate (but not ideal) cation selectivity in NaCl or KCl gradients. Other selectivity measurements suggest that Ca2+ and Mg2+ are also permeable. The channels show a slight voltage sensitivity. The steady state conductance at--70 mV (side opposite venom) is about 5 times the conductance at + 70 mV. We suggest that these channels in the venom may evoke transmitter release at neuromuscular junctions either by (1) depolarizing the pre-synaptic terminal and thus opening voltage-dependent Ca2+ channels, or (2) directly allowing Ca2+ to enter the terminal. Black widow spider venom is known to produce similar effects on neuromuscular junctions and lipid bilayers. The single channel conductances and ionic selectivities of the channels found in the venoms of Glycera and Latrodectus are strikingly similar. Taken together, these results suggest that channel formation can explain the electrophysiologic effects of these two different venoms. PMID- 6294924 TI - Central toxic effects of chronic ethanol treatment: actions on GABA and benzodiazepine recognition sites. AB - Prolonged ethanol treatment modifies various neurotransmitter systems. GABAergic neuronal function was particularly affected. On the other hand, clinical reports have indicated an interaction between ethyl alcohol and benzodiazepine receptors. These observations suggest a possible site of action of ethanol at the level of the GABA-benzodiazepine receptor complex. Our results showed that ethanol treatment differentially affected GABA receptor function and benzodiazepine binding sites. When [3H]GABA binding in the cerebellum, striatum and hippocampus was increased, [3H]diazepam binding remained unchanged in the same areas. The possibility of modulation of ethanol effects on GABAergic neurons through benzodiazepine receptors is discussed. PMID- 6294923 TI - Effect of nitrogen dioxide exposure on cyclic GMP in rat lung. AB - In vivo exposure of rats to 10 ppm nitrogen dioxide (NO2) for 6 h caused approx. 5-fold increase in the content of cyclic GMP in lung tissue. This increased cyclic GMP level lasted up to 24 h but returned to the normal level within 2 days, even when the NO2 exposure continued. This increase in the content of cyclic GMP of lung tissue by NO2 exposure was observed in both young and aged rats. There were no statistically significant changes in the content of cyclic AMP in lung tissue. PMID- 6294925 TI - Clinical study on etiologic aspects of primary hepatocellular carcinoma. AB - In spite of many animal experiments and clinical observations, the etiologic aspects of primary hepatocellular carcinoma remain obscure. In 20 cases of primary hepatocellular carcinoma occurring in cases of liver cirrhosis, the author clarified the etiologic aspects. In 10 out of 20 cases of primary hepatocellular carcinoma, an early history of blood transfusion could be detected. In eight cases, an early history of acute viral hepatitis including three cases of posttransfusion hepatitis could be clarified. In the five remaining cases, etiologic factors were unknown but an early history of anicteric hepatitis could not be ignored. In four out of 20 cases, HBs Ag in the serum could be detected. In spite of these findings, the close relationship between hepatitis virus infection and the pathogenesis of primary hepatitis virus infection and the pathogenesis of primary hepatocellular carcinoma must be taken into consideration. The period of time from blood transfusion and/or the onset of acute viral hepatitis to initial diagnosis of primary hepatocellular carcinoma appears to be reasonable for the occurrence of the disease. The question remains open as to what role hepatitis virus plays in the pathogenesis of primary hepatocellular carcinoma. PMID- 6294926 TI - [Physiological and pharmacological importance of presynaptic beta-adrenoceptors in modulation of noradrenaline release]. PMID- 6294927 TI - [Cardiac adrenoceptors: characterisation and regulation]. PMID- 6294928 TI - [Lymphography and a confusional syndrome]. PMID- 6294930 TI - Cholecalciferol sulfate identification in human milk by HPLC. AB - Synthetic vitamin D3 sulfate was prepared by reacting cholecalciferol with sulfamic acid in pyridine. Vitamin D3 sulfate ammonium salt was purified by crystallisation and transformed in sulfate sodium salt. Homogeneity was controlled by reverse phase high pressure liquid chromatography (HPLC). Purified synthetic vitamin D3 sulfate sodium salt was used as a reference. Milk whey was obtained after protein precipitation by adding ethanol. Vitamin D3 sulfoconjugate was identified in supernatant (lyophylized) after purification by Sephadex LH 20 and HPLC. Milk whey purified fraction obtained exhibited the same ultra-violet absorption (UV) as synthetic vitamin D3 sulfate; after solvolysis, cholecalciferol was liberated from natural and synthetic sulfoconjugate. The results confirmed that vitamin D3 sulfate was present in human milk. PMID- 6294929 TI - [Radiation exposure and radiation risk from natural radionuclides in the air]. PMID- 6294931 TI - [Myoblastoma: reference cases]. PMID- 6294932 TI - Possible relation of seropositivity to Japanese encephalitis in pigs with the 1978 epidemic of human encephalitis in Bihar. PMID- 6294933 TI - Quo vadis macrophage activation--role of phospholipids in the elicitation of the oxidative burst in macrophages. PMID- 6294934 TI - A rapid semi-quantitative glucose-6-phosphate dehydrogenase screening test for the clinical laboratory. PMID- 6294936 TI - Mammary tumors in BALB/cfC3H and BALB/cfRIII virgin female mice bearing hypophyseal isografts. AB - The behavior and morphology of mouse mammary tumors in high cancer strain mice are controlled by the causative murine mammary tumor viruses (MuMTV). However, relationships also exist between mammary tumors and other factors such as host genotype, other etiologic agents (chemicals, X-rays), and hormones. Among the latter, prolactin has been particularly studied. The aim of this investigation was to test whether the mammary tumor morphology controlled by C3H and RIII MuMTVs is in any way influenced by continuous prolactin secretion. A total of 28 BALB/cfC3H and 24 BALB/cfRIII virgin female mice was implanted with 2 hypophyseal isografts under the kidney capsule. Mice were of the same BALB/c genotype and carried respectively C3H and RIII MuMTV infection. The biologic and morphologic characterization of the tumors observed showed that several differences previously reported between mammary tumors in breeding female mice of the 2 strains persisted in virgins bearing hypophyseal isografts. These data confirm that the mammary tumor characters in high cancer strain mice are mainly carried out and controlled by the causative MuMTVs but also suggest that different MuMTVs may show different sensitivity to continuous prolactin secretion. PMID- 6294935 TI - Methodologic problems encountered in the assay of proteinases in Lewis lung carcinoma, a mouse metastasizing tumor. AB - The proteolytic activity in homogenates and extracts of subcellular fractions prepared from subcutaneous Lewis lung carcinoma was determined using proteins and synthetic peptides as substrates. The presence of cathepsin D, plasminogen activator, cathepsin B-, cathepsin G- and elastase-like enzymes was observed. No difference was revealed between the proteolytic activity in homogenates of Lewis lung carcinoma, at the growth stage examined, and in homogenates of normal lung. High specific activities were found in the lysosomal extract, whereas decreasing activities were found in the nuclear extract, the homogenate and the postlysosomal mitochondrial supernatant; no active or trypsin-activatable collagenase activity was detected. The presence in the tumor tissue of these enzymatic activities is in agreement with their proposed role in the process of metastasis. The lack of differences between homogenates of tumor and normal lung tissue suggests that the use of whole cells is required to selectively study tumor proteinases specifically involved in tumor malignancy. PMID- 6294937 TI - Effects of opiates and naloxone on certain enzymes of carbohydrate metabolism in human breast carcinomas. AB - The effects of administration of opiates and naloxone on the activities of PFK, 6PGDH and alpha-GPDH and alpha-GPDH/6PGDH ratios in human breast carcinomas were investigated in patients awaiting mastectomy. Injection of naloxone or fentanyl into an antecubital vein resulted in a statistically significant reduction in the activity of alpha-GPDH. Fentanyl was also effective in reducing the activity of 6PGDH. Injection of morphine into a branch of the internal mammary artery during mastectomy failed to induce changes in the activities of any of the enzymes but injection of naloxone resulted in a significant rise in the activity of 6PGDH. It is postulated that these alterations in the activities might not be associated with the binding of these drugs to opiate receptor proteins in the carcinoma. Furthermore opiate agonists or antagonists might not produce the required changes in the activities of any of the enzymes. PMID- 6294938 TI - [Treatment of irritable bowel with dietary fiber. A controlled clinical study]. PMID- 6294939 TI - [Epidural and intrathecal opiates]. PMID- 6294940 TI - [Epidural morphine]. PMID- 6294941 TI - [Diet for diabetics]. PMID- 6294942 TI - [Metabolic alkalosis in a newborn infant caused by topical application of bicarbonate]. PMID- 6294943 TI - Applications of nuclear medicine in genitourinary imaging. AB - Major advances in nuclear medicine instrumentation and radiopharmaceuticals for renal studies have occurred during the last decade. Current nuclear medicine methodology can be applied for accurate evaluation of renal function and for renal imaging in a wide variety of clinical situations. Total renal function can be estimated from the plasma clearance of agents excreted by glomerular filtration or tubular secretion, and individual function can be estimated by imaging combined with renography. A major area of radionuclide application is in the evaluation of obstructive uropathy. The introduction of diuretic renography and the use of computer-generated regions of interest offer the clinician added useful data which may aid in diagnosis and management. Imaging is of proven value also in trauma, renovascular hypertension, and acute and chronic renal failure. Methods for the evaluation of residual urine, vesicoureteral reflux, and testicular torsion have achieved increasing clinical use. These many procedures assure a meaningful and useful role for the application of nuclear medicine in genitourinary imaging. PMID- 6294944 TI - Application of pattern recognition techniques to breast cancer detection: ultrasonic analysis of 100 pathologically confirmed tissue areas. PMID- 6294945 TI - Wilms tumor in adult patient. PMID- 6294946 TI - Retroperitoneal and inguinal manifestation of malignant fibrous histiocytoma. PMID- 6294947 TI - Melanotropic effects of benzodiazepines: correlation with high-affinity receptors. PMID- 6294948 TI - Prolonged intracerebral infection with poliovirus in asymptomatic mice. PMID- 6294949 TI - Benign infantile mitochondrial myopathy due to reversible cytochrome c oxidase deficiency. PMID- 6294950 TI - In McArdle disease, phosphorylase deficiency is the tip of an iceberg. PMID- 6294951 TI - Age-dependent in vitro restriction of mouse neurotropic retrovirus replication in central nervous system-derived cells from susceptible Fv-1nn mice. PMID- 6294952 TI - Isolation of herpes simplex virus type 1 in recurrent (Mollaret's) meningitis. PMID- 6294953 TI - A summary of complications of fluid therapy. PMID- 6294954 TI - Electron microscopy of radiating clubs in tonsillar crypts of swine. AB - Tonsils of 25 adult swine with inflamed crypts were screened for the presence of radiating clubs, and five were selected for electron microscopic study of pathogenesis of clubs. Radiating clubs in crypts were surrounded by an exudate which principally contained neutrophils. Clubs were present on the outer surface of bacterial microcolonies; they also were present on the outer surface of plant particles wedged in crypts or on the cell walls of these particles near bacterial microcolonies. The study demonstrated development of clubs from an amorphous material derived from bacterial and leukocytic cells and also from precipitated proteins. The amorphous material was shaped into clubs by a protracted phagocytic activity of numerous neutrophilic leukocytes. The lesions with radiating clubs within crypts were not surrounded by granulomatous inflammation due to the intact epithelial lining of crypts, and therefore they differ from the granulomas having clubs around microbial colonies. They could, however, serve as models of pathogenesis of radiating clubs in the purulent core of specific granulomas with radiating clubs. PMID- 6294955 TI - Rhinomune (rhinopneumonitis vaccine) PMID- 6294956 TI - Distribution of hexoestrol residues in caponised chickens. AB - A radioimmunoassay method is described for the detection and measurement of residues of hexoestrol and other stilbenes in tissues of poultry. The residues following caponisation with 12 mg hexoestrol were measured in leg muscle, liver, visceral fat and neck 44 days after implantation. They were significantly greater than values obtained from eight untreated control birds. The mean values obtained in the caponised chickens ranged from 471 pg/g net weight of leg muscle to 584,500 pg/g of tissue from the upper neck region, which included the site of implantation. Control values in untreated birds fell within the range of 8 pg/g in leg muscle to 44 pg/g in liver tissue. PMID- 6294957 TI - Welfare in veal calf units. AB - Compared to France, Italy, the Netherlands and Germany the production of white veal in the United Kingdom is relatively limited. Nevertheless, public interest in veal calf welfare appears to be lively. Although calves are sucklings of social-living ruminants, veal calves are not allowed to suck, to have a social life or to ruminate. This, added to anaemia in order to obtain white meat, and the high rate of morbidity caused by high density of the animals, sums up the welfare problems which arise. In principle, there are two types of housing: single housing in crates or group housing. Group housing is almost impossible for calves younger than six weeks, because of disease control and urine sucking. Even after this age it requires much skill and labour. Enforceable regulations within the European Economic Community concerning the amount of light in the fattening houses and the measurements of crates for individual housing are suggested. It is recommended that anaemia in young calves should be treated. PMID- 6294958 TI - Blood dried on filter or blotting paper for the detection of antibody against swine vesicular disease virus by enzyme-linked immunosorbent assay. PMID- 6294959 TI - A common code of practice for the control of contagious equine metritis and other equine reproductive diseases for the 1983 covering season in France, Ireland and the United Kingdom. PMID- 6294960 TI - Herd eradication of enzootic bovine leukosis. PMID- 6294961 TI - From firing to phenylbutazone in equine practice. PMID- 6294962 TI - Electrophoretic comparison of the genomes of North American bluetongue viruses, one Australian bluetongue virus, and three other related orbiviruses. AB - The genomes of U.S. bluetongue viruses, an Australian bluetongue virus, and three other related orbiviruses were analyzed by polyacrylamide gel electrophoresis. The genomes were comprised of ten segments of double-stranded (ds) RNA. Estimates of the molecular weights of the dsRNA segments revealed that the U.S. bluetongue serotypes were remarkably similar. Although the dsRNA profiles of the viruses exhibited common segments, each virus had a distinct dsRNA profile. The usefulness of the genome analysis as a diagnostic tool for identification and for epidemiologic studies is discussed. PMID- 6294963 TI - Experimental infection of lambs with an adenovirus followed by Pasteurella haemolytica. PMID- 6294964 TI - Humoral and cell-mediated immune responses in chickens with infectious bursal disease. AB - Primary and secondary immune responses to Newcastle disease virus (NDV) was evaluated in chickens infected with infectious bursal disease virus (IBDV) at one and 28 days of age. The geometric mean primary hemagglutination-inhibition antibody titers (GMT) of chickens infected with IBDV at one day of age was significantly lower (P less than or equal to 0.01) than those infected at 28 days of age. Infection with IBDV had no influence on secondary immune response to NDV. The effect of IBDV infection at one day of age on the cell-mediated immunity of chickens was evaluated by skin allograft acceptance or survival time. There was no significant difference between the percentage of grafts accepted in IBDV infected and noninfected control chickens. However, the mean graft survival time in the IBDV infected chickens was significantly longer (P less than or equal to 0.05) than those in the control group. This suggested a suppression of cell mediated immunity due to IBDV infection. PMID- 6294965 TI - Analysis of buoyant density of canine peripheral blood leukocytes with PVP-Silica (Percoll) density gradients. AB - Isolation of canine peripheral blood mononuclear cells with a one step centrifugal separation procedure has not been very successful sofar. Significant contamination with polymorphonuclear cells has been reported. An analysis of the buoyant density of canine peripheral blood leukocytes on a self-generating Percoll gradient showed that the buoyant densities of polymorphonuclear cells and lymphocytes are so near that separation with high purity and yield is not possible with the use of a density gradient. Transient changes in buoyant density of polymorphonuclear cells have been observed. In such situations differences in buoyant density between cell types have been observed which permit separation of mononuclear cells from polymorphonuclear cells at a reasonable yield. PMID- 6294966 TI - Protection against bovine leukosis virus infection in sheep with the BL 20 bovine lymphoblastoid cell line. AB - The bovine lymphoblastoid BL 20 cell line derived from a case of sporadic bovine leukosis when inoculated into sheep did not induce an antibody response directed against bovine leukosis virus (BLV) structural proteins. Sheep were inoculated twice with the BL 20 cell line and then challenged with BLV infected lymphocytes. Three out of four sheep challenged four weeks after BL 20 inoculation did not develop BLV antibodies. Of the 12 sheep challenged later, three sheep did not develop BLV antibodies. BLV was isolated from all the seropositive animals and from none of the seronegative animals. PMID- 6294967 TI - [Adaptation of attenuated strain of P. parainfluenzae-3 in cultured swine kidney cells and analysis of its immunogenicity]. AB - Strain Mihailovgrad of P. parainfluenza-3 was successfully adapted to cell cultures of pig kidney at its 15th passage. Its testing for innocuity was carried out through the inoculation of test calves with high amounts of the pathogen. No pathologic, temperature, and anaphylactic reactions were established. This confirmed the harmless character of the attenuated strain. Comparative testing for immunogenicity was performed with 8-month-old calves. Full coincidence with the antibody response was obtained with strain Mihailovgrad as a vaccinal strain. The use of the adapted strain for the production of a vaccine is to be preferred as there is no possibility of contamination with other bovine viruses. PMID- 6294968 TI - [Serological study of the bovine virus diarrhea-mucosal disease]. AB - Employed were the virus-neutralization test (VNT), the complement-fixation tests (CFT), and the immunodiffusion test (IT) in the study of serum samples of calves on a total of 25 farms that had records of virus diarrhea-mucosal disease (VD-MD) in the course of two years. Samples were also taken from calves in the initial phase of the same disease in the course of four months. The blood of experimentally infected calves waw likewise sampled for three months. It was found that the stage of infection could successfully be studied via all three serologic methods. Thus, for example, positive VNT and CFT results coupled with negative IT results spoke of the initial stage of the disease, and vice versa- negative CFT results and positive VNT and IT results gave evidence of the advanced stage of the infection. PMID- 6294969 TI - [Detection of antibodies against bovine leukemia virus using the enzyme-linked immunosorbent assay]. AB - Described is the experimental procedure of the enzyme-linked immunosorbent assay (ELISA) in the detection and quantitative determination of bovine leukosis virus antibodies. The optimum conditions for the employment of the method are stated along with the optimum amounts of the reagents taking part in the performance of the assay. It was established that as much as 10 ng/cm3 virus purified in gradients was sufficient to sensitive the polyvinyl plaques. A comparative investigation was carried out through the agar gel immunodiffusion test, using a double antigen. Forty-seven out of a total of 446 sera were found to be ELISA positive, and 45 were positive as demonstrated by the immunodiffusion test. Results indicated that ELISA could be used as a routine method in the diagnosis of enzootic bovine leukosis. PMID- 6294971 TI - [Adenovirus infections of cattle]. AB - Results are given of studies on the adenoviral infections in cattle in the region of Southeast Bulgaria. Serologic methods were used to elucidate some moments of the enzootic process and the part played by bovine adenoviruses in the etiology of the respiratory diseases of calves. Demonstrated was the participation of adenovirus strains of group II. It was established that the technologic moments influence the course of the adenoviral enzootics. A successful attempt was made to uncode the type picture and the circulation of types 1, 2, 3, and 8 adenoviruses within the cattle herds. The method of direct immunofluorescence was successfully employed in the diagnostics of adenoviral diseases in calves when alive. Demonstrated was also the presence of specific skin supersensitivity in cattle infected with adenoviruses. Data are given on the specificity of the reaction, the quality and standardization of the allergen, the age-associated dynamics of the process, etc. PMID- 6294970 TI - [Bulgarian potassium-calcium zeolite in the prophylaxis of digestive tract disorders in calves]. AB - Scientific and economic trials were carried out to test the prophylactic effect of Bulgarian K-Ca zeolite products in the prevention of digestive troubles in calves, corresponding to the State Standard production branch requirements of 33675789-79, and to 0.00-0.01 mm (France). When applied at the rate of 2 per cent constituent of the daily milk ration from the first to the fifteenth day after calving it was found to produce a good prophylactic effect against digestive disturbances. It was also found that there was enhancement of the general resistance in calves. This resulted in lower morbidity and mortality rates, and shortening the period of illness in calves that received zeolite. By the end of the experimental period there was a definite rise of the blood albumins, the sum total of the gamma-globulins, the hemoglobin value, and the absolute values of granulocytes and lymphocytes. PMID- 6294972 TI - [Relationship between the genetic polymorphism of alkaline phosphatase and the resistance of broiler poultry to Marek's disease virus]. AB - Studied was the polymorphism of alkaline phosphatase in blood plasma samples taken from 507 poulets and cocks of lines 66 and 77 of the Cornish breed, and lines 88 and 99 of the White Plymouth Rock breed. The birds were divided into three groups to test their resistance to Marek's disease: (1) controls, (2) infected with the virus of Marek's disease with 3 subgroups--resistant birds, slightly susceptible ones, and strongly susceptible ones, and (3) contacts, with two subgroups--survivals and birds that died. The blood plasma alkaline phosphatase was determined through horizontal electrophoresis on a starch gel, employing the method of Gahne. It was found that the alkaline phosphatase genetype FF frequency was lower with lines 66 and 77 than with line 88 and especially with line 99 of the White Plymouth Rock breed which proved most susceptible to the virus of Marek's disease. In the homozygotic alkaline phosphatase genetype SS none of the investigated lines showed any mortality. The resistance of broiler birds to Marek's disease was governed by the Akps allele, while the susceptibility was governed by the Akpf one. The former could be used as a genetic marker in the selection of initial lines of broiler birds by resistance, and the latter could be used as a marker in the selection by susceptibility. PMID- 6294973 TI - [Breed, line and sex differences of phagocytic activity in relation to broiler poultry resistance to Marek's disease virus]. AB - The phagocytic activity was studied of 421 poulets and cocks of lines 66 and 77 of the Cornish breed, and lines 88 and 99 of the White Plymouth rock breed. Breed and linear differences were noticed in the changes of the phagocytic activity (phagocytic number and phagocytic index) with birds infected with the virus of Marek's disease. The rise of the phagocytic activity with lines 66 and 77 was slighter as compared with that of the White Plymouth Rock lines 88 and 99. Strongest were the changes in the phagocytic activity with birds that were strongly susceptible to Marek's disease, and slight est--with birds that were resistant to the disease. A trend of a better expressed phagocytic activity was noticed with the cocks of all investigated lines. PMID- 6294974 TI - Microvasculature of human colorectal epithelial tumors. An electron microscopic study. AB - The ultrastructure of the microvasculature in human colorectal adenomas and carcinomas was studied, and compared with that of normal tissue. Blood vessels in adenomas were generally of normal structure, whereas carcinoma vessels displayed a variety of structural alterations. Endothelial proliferation was frequently observed in all the eleven carcinomas examined, which confirms the reports of new vessel formation in experimental malignant tumors. The presence of fenestrations in obviously abnormal endothelium was tentatively attributed to hypoxia, vascular regression or immaturity of the cells, in spite of the fact that normal capillaries in colonic mucosa are fenestrated. Some vessels showed multilayered basement membranes, various types of activated cells containing numerous lysosomal granules, and thickened perivascular tissue. These features may be a vascular response to repeated damage or a manifestation of vascular remodelling. Dilated thin-walled vessels, which were usually found at the periphery of carcinomas, were identified as venules. Small vessels located between the closely apposed carcinoma tubules were not obviously abnormal. They were nevertheless considered to be tumor-induced vessels which had undergone differentiation. The differences in vascular morphology between adenomas and carcinomas were considered to be due to their different growth patterns, growth rates and degrees of maturity. PMID- 6294975 TI - Demonstration of alpha 1-antitrypsin in paraffin sections of hepatoma and cirrhosis. AB - Alpha 1-antitrypsin has been examined in formalin-fixed, paraffin-embedded liver specimens from Greek patients with cirrhosis (35 cases) and hepatoma (55 cases) by peroxidase-antiperoxidase (PAP) method. Ring-like AAT globules were found in the non-neoplastic cells in 12% of the cases of hepatoma and in 11% of the cases of cirrhosis. Atypical globules were seen in neoplastic cells in 5.4% of the cases of hepatoma and in 17% of the cases of liver cirrhosis. A diffuse fine granular pattern of AAT distribution was present in 31% of the cases of hepatoma in the neoplastic cells and in 27% of those in the non-neoplastic cells. The relatively high incidence of ring-like AAT-globules, and of atypical globules in cases of hepatoma and cirrhosis is not in agreement with the extremely low gene frequency of Z allele in a Greek population of patients with cirrhosis and hepatoma. Thus, there is some doubt whether AAT-globules in the liver represent a histopathologic marker of genetically determined AAT deficiency. A relationship between AAT deposits and the degree of differentiation of hepatoma was noted in this series. AAT-positive cells were found in 55% of moderately differentiated, in 29% of highly differentiated and in 20% of poorly differentiated hepatomas. PMID- 6294976 TI - Distribution of epithelial membrane antigen in benign and malignant lesions of the salivary glands. AB - An antiserum against epithelial membrane antigen has been used to stain a variety of lesions arising in the salivary glands. In normal major and minor glands staining was localised to the ductal systems. There was no evidence of myoepithelial cell staining. The mucous elements of the submandibular and sublingual glands were negative, but in the mucous elements of the minor glands there was focal cytoplasmic positivity. There was no cytoplasmic staining of serous elements in major or minor salivary glands. In pleomorphic adenomas the luminal membrane of ductal elements was strongly positive, with focal cytoplasmic positivity in some myxoid areas. In mucoepidermoid tumours both adjacent cell membranes and cytoplasm were strongly positive. The ductal structure of adenoid cystic carcinomas were clearly delineated while the pseudoducts produced by enclosed areas of stroma were negative. All mesenchymally derived tumours were negative and a tumour previously considered as a chondroma was strongly positive. The results are discussed in relation to phenotypic heterogeneity and the histogenesis of salivary gland tumours. PMID- 6294977 TI - Herpes simplex virus type 1 cell fusion occurs in the presence of ammonium chloride-inhibited glycoproteins. PMID- 6294978 TI - Isolation and characterization of four new temperature-sensitive mutants of avian erythroblastosis virus (AEV). PMID- 6294979 TI - Cell fusion induced by Nelson Bay virus. PMID- 6294980 TI - Characterization of P1argF derivatives from Escherichia coli K12 transduction. III. P1Cm13argF derivatives. PMID- 6294981 TI - Chromosomal organization of the herpes simplex virus type 2 genome. PMID- 6294982 TI - Polyoma virus-transformed cells produce transforming growth factor(s) and grow in serum-free medium. PMID- 6294983 TI - Synthesis of VSV RNPs in vitro by cellular VSV RNPs added to uninfected HeLa cell extracts: VSV protein requirements for replication in vitro. PMID- 6294984 TI - A study of rat embryo cells transformed in vitro by the bovine adenovirus type 3 (BAV-3) DNA before and after a passage in the host. PMID- 6294985 TI - Nucleotide sequence analysis of the recombinant joints in 16 naturally arising deletion mutants of simian virus 40. PMID- 6294987 TI - Concatemers in a rapidly sedimenting, replicating bacteriophage T7 DNA. PMID- 6294986 TI - Synthesis of vaccinia virus thymidine kinase in microinjected Xenopus oocytes. PMID- 6294988 TI - T4 bacteriophage-coded polynucleotide kinase and RNA ligase are involved in host tRNA alteration or repair. PMID- 6294989 TI - Cyclic nucleotide levels in light- and dark-adapted ground squirrel whole eyes. PMID- 6294990 TI - [Modern concepts of the pathogenesis of alcoholism]. PMID- 6294991 TI - [Clinical aspects and diagnosis of complications of diphtheria of the oropharyngeal isthmus in adults]. PMID- 6294992 TI - [Role of phosphodiesterase in the changes of cyclic adenosine-3',5'-monophosphate in the rabbit jejunal mucosa after treatment with cholera enterotoxin]. AB - An increase in content of cAMP in the rabbit jejunum mucosa after treatment with cholerae toxin was due to adenylate cyclase activation. The maximal activation of adenylate cyclase (AC) and increase in the cAMP content in the mucosa were observed within 4 hrs after administration of the cholerae toxin in situ into the lumen of an isolated strip of the rabbit jejunum. Cholerae enterotoxin did not affect the activity of phosphodiesterase (PDE). It was shown that AC activation did not correlate with an increase in the cAMP content in rabbit jejunum mucosa exposed to cholerae enterotoxin. Absence of the correlation was demonstrated by kinetic characteristics of PDE and by regulatory effect of Ca2+ on the activity of AC and PDE. PMID- 6294994 TI - [Effect of diets with different content of polyunsaturated fatty acids on ATPase activity in different rat tissues]. AB - The authors studied the effect of the diets with varying content of polyunsaturated fatty acids (PUFA) on ATPase activity in the heart, kidneys and lungs of Wistar rats. During 3 weeks the rats received different diets. Group I was kept on the diet poor in PUFA, Group II on the diet rich in linoleic acid, and Group III on the diet rich in linolenic acid. The content of PUFA in food has no effect on ATPase activity in the lungs. The maximal activity of Mg-ATPase in the heart was seen in the Group III rats, while the minimal in the Group I rats (P less than 0.05). On the contrary, Na, K-ATPase activity was minimal in the Group III rats (P-0.05). Examination of ATPase activity in the kidneys has demonstrated no substantial difference between Groups II and III as regards Mg ATPase activity, whereas Mg-ATPase activity in the Group I animals was significantly reduced (P less than 0.02). As Na, K-ATPase activity in the kidneys increased (Group II, P-0.05) excretion of Na and K with urine considerably diminished. PMID- 6294993 TI - [Interaction of various tetrahydroisoquinoline alkaloids with opiate receptors in the rat hypothalamus and midbrain]. AB - Using the method of competitive displacement of 3H-labelled naloxone, (D-ala2) met enkephalinamide or (D-ala2)-D-leu-enkephalin by salsolinol, 1-methyl-6 hydroxy-1, 2, 3, 4-tetrahydro-beta-carboline or 1-methyl-6-methoxy-1, 2, 3, 4 tetrahydro-beta-carboline from opiate receptors, it was shown that the alkaloids studied were capable to cause specific interactions with rat hypothalamus and midbrain however, exhibiting distinctly less affinity as compared with morphine or its analogs. The sodium ratios, determined from the effective doses of the tetrahydroisoquinoline alkaloids corresponding to the alkaloid concentrations, which induce 50% displacement of 3H-naloxone from the opiate receptors in the presence or absence of 100 mM NaCl, have been found to be 0.75 for salsolinol and 3.6 for beta-carbolines studied. The data obtained suggest that salsolinol, similar to naloxone, is a "pure" morphine antagonist, whereas the beta-carbolines studied may be classified with the agonist-antagonist type. A considerable decrease in the affinity of mu-type opiate receptors has been found in presence of salsolinol in the incubation medium. The possible mechanisms of pharmacological action of the alkaloids and their relation to development of alcohol dependence and tolerance are discussed. PMID- 6294995 TI - Cryopreservation of platelets isolated with the IBM 2997 blood cell separator: a rapid and simplified approach. AB - The equivalent of 5-7 units of platelets, isolated from a single donor with the IBM Blood Cell Processor 2997 using a dual stage separation chamber, was frozen with the cryoprotectant dimethylsulfoxide (DMSO). The DMSO-saline solution was added directly to the platelets, and the platelets were frozen in a polyvinyl chloride plastic bag by storage in a -80 degrees C mechanical freezer. Washing the thawed platelets with a phosphate-buffered sodium chloride-dextrose solution, pH of 5.0, removed about 95% of the DMSO. In vitro freeze-thaw-wash recovery was 80%, and in vivo 51Cr platelet recovery was 31%. Platelet dense body granules were well maintained after freezing, thawing, and washing. This is a safe and effective method of platelet cryopreservation which can be performed in less time than other currently used methods. PMID- 6294997 TI - [Calcium antagonists in the treatment of cardiovascular diseases (review of the literature)]. PMID- 6294996 TI - [Hormone secretion in hypophyseal adenomas]. PMID- 6294998 TI - [Primary malignant histiocytoma of the heart]. PMID- 6294999 TI - [Catecholamines in the therapy of chronic cardiac insufficiency]. PMID- 6295000 TI - The management of the hypoglycemic patient. PMID- 6295002 TI - Chronic congenital infections of man. AB - This discussion focuses on the dynamic nature of three chronic congenital infections of man, namely those caused by cytomegaloviruses, rubella virus, and Toxoplasma gondii. The spectra of the fetal infection are described and the risks associated with the acute and chronic phases of the fetal infections are contrasted. Because of their frequency and subtlety, emphasis is placed on the late-appearing sequelae which are associated with the persistent and/or recurring aspects of these infections. PMID- 6295001 TI - The radiologist's role in acute endocrine problems. PMID- 6295003 TI - The clinical illness promotion factor: a third ingredient. AB - The interactions between a causative agent and a susceptible host involve a series of responses most of which are subclinical or asymptomatic but a few of which are manifested by clinical illness. The factor(s) which tip the balance are poorly understood in both acute and chronic diseases. It is designated here as the clinical illness promoting factor (CIPF), a third ingredient. Among infected persons some leads have been found as to why clinical illness develops: in tuberculosis genetic susceptibility plays a key role, as shown in twin studies; in EBV infections age at the time of infection, genetic, and psychosocial factors determine both the expression and the severity of illness; in poliomyelitis age, exercise in the incubation period, and genetic background are related to the development of paralysis. In the relationship between viruses and cancer, viruses and chronic diseases, or inanimate pathogens like tobacco and lung cancer, we know very little as to the factors that result in clinical disease among the many who are presumably susceptible and fully exposed. Epidemiologic study is urged to identify this CIPF or "third ingredient." PMID- 6295005 TI - The enteroviruses: recent advances. AB - New information accrues periodically in the ledger accounting for infections and diseases associated with the human enteroviruses. The discoveries of "new" serotypes and how they affect people are subjects of continuing attention. Some other relevant information on "old" serotypes relates to variations in age specific attack rates and associated morbidity and mortality for neonates and older infants. Among the morbidity reports are recounts during outbreaks of virus positive cerebrospinal fluids (CSFs) that initially may not have cytological or biochemical abnormalities. Prolonged enterovirus infections may develop in persons having agammaglobulinemia. Lastly, some provocative associations concern the pathologic expressions of enteroviruses in the development and persistence of injury to the heart (myocardiopathies) and the pancreas (insulin-dependent diabetes mellitus). PMID- 6295004 TI - Infectious mononucleosis: observations on transmission. AB - Epsten-Barr virus oropharyngeal shedding has been demonstrated in infectious mononucleosis patients many months after acute illness and long after the disease hallmarks, atypical lymphocytes and heterophile antibody, have disappeared. Extracellular virus is present more frequently in saliva than in other oropharyngeal samples. Prolonged excretion of EBV in asymptomatic carriers explains the difficulty in tracing case-to-case spread and increased transmissibility in age groups in which salivary exchange is high. PMID- 6295006 TI - Prospects for human monoclonal antibodies: a critical perspective. AB - Monoclonal antibodies have proved useful in detecting antigenic variation at single determinant sites on complex antigens, a fact which seems to have engendered a common misperception that a monoclonal antibody is necessarily a "monospecific" agent. Yet there exists considerable evidence that individual antibody molecules are multispecific in their binding capabilities, and that the amount of functional and genetic redundancy in mammalian immune systems may be quite large. The implications of this data for the future of monoclonal antibodies are emphasized, along with a brief review of present progress toward human monoclonal antibodies. PMID- 6295007 TI - Immortalization of human lymphocytes by Epstein-Barr virus. AB - Epstein-Barr virus (EBV) confers upon normal lymphocytes derived from bone marrow the ability to proliferate indefinitely in a test tube. This process, called immortalization, is crucial to the pathogenesis of EBV infections. Inside the immortalized lymphocyte the EBV genome exists as a complete multicopy circular plasmid which is probably not integrated into the cell chromosome. Most of the viral genetic information is not expressed. However, at least six to eight separate regions of the EBV genome encode viral products which are made in the immortalized cell. The identification of the function of these few genes holds some interesting answers to questions concerning the biochemical mechanisms of control of lymphocyte growth and differentiation. PMID- 6295009 TI - Chickenpox encephalitis and encephalopathy: evidence for differing pathogenesis. AB - Retrospective assessment of hepatic and central nervous system involvement associated with chickenpox cases at a large metropolitan medical center reveals that 28 of 58 patients had biochemical, but not inflammatory, evidence of liver involvement. An additional 18 patients had biochemical liver abnormalities along with non-inflammatory encephalopathy (Reye syndrome) and 12 had clear evidence of central nervous system inflammatory involvement (encephalitis). There were no cases of solitary inflammatory liver involvement. Reviewed evidence suggests that the pathogenesis of hepatopathy and hepatoencephalopathy (Reye syndrome) is not caused by replication of virus in the involved organs, but instead is mediated through a cytotoxic mechanism and that the inflammatory brain disease is also not caused by viral replication in brain tissue, but appears to be tissue damage associated with immune cell responses (post-infectious encephalitis). The concept put forth in this essay is that a virus replicating in one organ (skin) could affect the macromolecular function of cells in another organ (liver, brain) bringing about both hepatopathy and hepatoencephalopathy. PMID- 6295010 TI - The functions of a university-based viral diagnostic laboratory: recent experiences at Yale-New Haven Hospital. AB - The work of the Viral Diagnostic Laboratory at Yale-New Haven Hospital has been continuously inspired by the virologic and epidemiologic principles taught us by Dr. Horstmann. This paper helps to define the various functions of this facility. Perhaps its most important role is as a learning laboratory where generations of students, fellows, house officers, and teachers are taught how viruses behave in the human host and in the community. PMID- 6295008 TI - The biology of circulating B lymphocytes infected with Epstein-Barr virus during infectious mononucleosis. AB - EBV-infected B cells are present in blood during acute infectious mononucleosis (IM) in surprisingly large numbers. These cells share with cells transformed by EBV in vitro the capacity for unlimited proliferation, but this capacity is quite restricted in the normal host. The unusual extent of plasmacytic differentiation seen in infected cells in vivo early in IM and the loss of this differentiation in the late stages of the disease suggest that these cells are subject to the same immunoregulatory mechanisms that affect normal B cells. The host response to virus-altered B cells is quite complex but is perhaps best viewed as an in vivo autologous mixed lymphocyte reaction initiated by the interaction of T cells with virus-activated B cells. PMID- 6295011 TI - New opportunities for development of safe, effective live virus vaccines. AB - Effective vaccines are not available for most viral diseases. This situation may soon change when the full force of contemporary molecular biology is applied to immunoprophylaxis. In certain viral diseases, particularly those affecting the respiratory and gastrointestinal tracts, live attenuated vaccines are needed to confer effective protection. Until now the major obstacle to success has been genetic instability. It may be possible to construct stable, satisfactorily attenuated mutants by cloning viral DNA or RNA and then subjecting the cloned DNA to enzyme surgery to create viable deletion mutations. Modified cloned DNA derived from positive-strand viral RNA could then be transferred back into its virus by transfection of cells. Conversion of mutant cloned DNA into negative strand RNA and transfer into its virus will require a more elaborate type of rescue. PMID- 6295012 TI - Antiviral chemotherapy--a frontier for health and learning. AB - Antiviral chemotherapy has been too long perceived as being relatively impossible. Such notions adversely affect the acquisition of important specific clinical information, whereas much new knowledge is available about viral replication and cell biology which enhances the prospects for effective chemotherapy. Some immediate goals can be recognized that will further determine the ability to influence viral infections and properly interpret the drug effects. In recent controlled observations there is reason for expectant optimism, but the demonstration of antiviral chemotherapy is both disease- and host-dependent, with important nonpharmacologic aspects. Rapid specific and sensitive diagnostic tests are of paramount importance; that they can be devised is a generally accepted conclusion among virologists. Problems in the scientific evaluation of antiviral chemotherapy in man have led to the recommendations of compounds that have no proved effect; amantadine, Ara A, and interferon, however, have been shown to be efficacious. Acyclovir and bromvinyldeoxyuridine have demonstrated virus-directed chemotherapy with impressive specificity. The frontier of antiviral chemotherapy holds great promise for additional learning and improved health through the implementation of developing knowledge. PMID- 6295014 TI - [Cytomegaly]. PMID- 6295013 TI - Hepatitis virus vaccines: present status. AB - During the past decade there has been extraordinary progress toward the development of vaccines for the prevention of type A and type B hepatitis. The successful propagation of hepatitis A virus in cell culture in 1979 was followed by the preparation of experimental live attenuated hepatitis A vaccines that have been shown to induce antibody in marmosets and chimpanzees and protect immunized marmosets against challenge with hepatitis A virus. The first human immunization trials will begin in mid-1982. An inactivated hepatitis B vaccine that was licensed in the United States in November 1981 has been shown to be safe, immunogenic, and effective. When this vaccine becomes available for use in July 1982, it will be recommended for persons who are considered to be at increased risk of contracting hepatitis B infection. Future generations of hepatitis B vaccines may be prepared from hepatitis B surface antigen derived from DNA recombinant technology or by in vitro synthesis of HBs Ag determinants by chemical means. PMID- 6295016 TI - [Electron microscope studies of lipid droplets in the amniotic fluid]. PMID- 6295015 TI - [Stevens-Johnson syndrome in childhood]. PMID- 6295017 TI - Neurosecretory and microstructural changes in the hypothalamus of rats following administration of lithium chloride and 3H-thymidin. AB - Adult male rats were intraperitoneally administered aqueous solution of lithium chloride (LiCl). Studies, including neurosecretory and microstructural changes within particular neurocytes in supraoptic (NSO) and paraventricular nuclei (NPV) were performed on hypothalamic sections. In the experimental rats the administered LiCl increased the level of GOMORI-positive neurosecretory material both in supraoptic and paraventricular nuclei. Great amounts of the neurosecretory material were markedly conspicuous in the above areas after 20 days of LiCl administration. Investigations carried out on cellular nuclei of particular neurocytes showed a significant enlargement of the nuclei, and statistical calculations revealed that, in comparison with the basic control, the difference was essentially significant (p less than 0.001). 3H-thymidin administration to the rats which had previously been on LiCl for 20 days demonstrated also that within supraoptic nuclei the incorporation of the isotope in cellular nuclei took a faster course than in control animals. PMID- 6295018 TI - [Ultracytochemical analysis of nuclear ribonucleoproteins]. PMID- 6295019 TI - [Structural bases of the reliability of cortical neuronal functioning]. PMID- 6295020 TI - [Physico-chemical mechanism of interaction of phagocytic cells with fibrosis inducing dusts]. PMID- 6295022 TI - Additional data on PGP genetic transmission in family groups. AB - PGP (phosphoglycolate phosphatase, EC 3.1.3.18.) gentic studies were performed in 188 families including a total of 415 offspring in the Galician population. The results are in agreement with the formal hypothesis of three codominant alleles at an autosomal locus. No silent alleles have been observed. PMID- 6295021 TI - A study of ten red cell enzymatic markers in the Naples' population. Report of a new GPT variant phenotype. AB - A sample of the population of Naples has been examined for several red cell enzyme markers. About 2,000 newborn have been analyzed for ACP, GLO I, and UMPK; 1,000 of them were also analyzed for PepA and PepB, and 500 for PGM1 and PGM2. In addition about 400 school children have been typed for the PGD and PGP polymorphisms. The observed gene frequencies for the polymorphic systems are: ACPA = 0.293, ACPB = 0.667 and ACPC = 0.040; GLO1 = 0.372; GPT2 = 0.462; UMPK2 = 0.029; PGM21 = 0.279; PGDC = 0.037; PGP1 = 0.953, PGP2 = 0.038 and PGP3 = 0.009. Moreover during the screening of PepA, PepB and GPT markers, some rare alleles have been encountered, one of which, at the GPT locus, has never been reported before. We propose for it the name GPT10. PMID- 6295023 TI - [Retrovirus activating potential of nuclear polyhedrosis viruses in mammalian cell cultures]. AB - Activation of endogenous C-type retroviruses by nuclear polyhedrosis viruses (NPV) was investigated in "in vitro" cell culture systems of four mammalian species: mouse, rat, monkey and man. Cells were treated with NPV, (isolated from larvae and insect cell cultures), NPV-DNA, C-type virus-activating chemicals (halogenated pyrimidine analogue) and chemical insecticides alone and in combination. C-type retroviruses were activated only in mouse cells and transformed rat cells by the halogenated pyrimidine analogue. In NPV-treated cell cultures no C-type retrovirus activation was detectable. In simultaneous treatments of the cells with nuclear polyhedrosis viruses and chemicals no potentiating effects by the NPV could be detected. Virions of NPV in cell cultures showed unaltered morphology and upon reisolation were infectious in homologous insect cell cultures for a long time. No influence upon growth or morphology of the treated mammalian cells could be observed. From our experiments we conclude that the application of nuclear polyhedrosis viruses for pest control is uncritical with respect to their retrovirus activating potential in mammalian cells. PMID- 6295024 TI - Experimental studies on enteropathogenicity of Vibrio cholerae serotypes other than 1. AB - Vibrio cholerae serotypes other than I have come to be recognised as an important aetiological agent of choleraic diarrhoea and a few of them have been shown to ben enterotoxigenic in experimental models. This study was taken up with strains belonging to all the known 59 serotypes. Live cells and culture filtrates of all the strains could cause accumulation of fluid in rabbit ileal loops. However, eleven strains did so after 1-2 serial passages in rabbit gut. All the culture filtrates could also cause increased vascular permeability. The toxin was found to be heat and pH labile. The pattern of time course of fluid accumulation in ileal loop and negative suckling mouse assay indicated that the enterotoxin was heat labile. Reduction of fluid outpouring by chlorpromazine but not by synthesis and receptor blockers of prostaglandin and 5-hydroxytryptamine indicated that the toxin may act through mediation of cAMP. All the 10 strains tested were negative in Sereny's test indicating lack of invasive capacity of these organisms. PMID- 6295025 TI - Rapid detection of Treponema pallidum and cytomegalovirus specific IgM antibodies with the passive haemagglutination. AB - For the detection of Treponema pallidum-specific IgM antibodies in patients' serum samples four simple and rapid methods are compared and evaluated. They show decreasing sensitivity and specificity in the row listed: (1) Treponema pallidum haemagglutination (TPHA) after IgM separation by affinity chromatography on controlled-pore glass heads (anti-IgM-CPG), or (2) by ion exchange chromatography on DEAE-cellulose; (3) solid-phase haemagglutination (SPHA) in glass microvials or (4) on polysterene microplates. The latter tests have found to be inferior, although they can be considerably improved by the use of affinity chromatography- purified catch antibodies. The commercially available DEAE-cellulose microcolumn delivers a simple method for IgM separation, and in spite of a relatively high IgA contamination it proved to be useful also for CMV-specific IgM haemagglutination tests, as compared to ELISA results. PMID- 6295026 TI - [Release of extracellular metabolic products by Streptococci groups C and G]. AB - 24 group C and 31 group G streptococcal strains were studied examining the production of extracellular nucleases, NADase, hyaluronidase and streptolysin O. 5 mucous strains were characterized by diminished growth and lack of hyaluronidase and NADase-production. Regarding the other strains of group C, one half produced nuclease-activity whereas only one out of ten group G streptococci did. Group G streptococci were more active in the production of NADase and hyaluronidase than group C streptococci. The levels of streptolysin O in groups C and G are comparable. PMID- 6295028 TI - [Experimental studies on extraosseous calcification in hypervitaminosis D3. I. Significance of proteoglycans in the early stages of calcification in the rat]. PMID- 6295027 TI - [Occurrence of IgM antibodies against cytomegalovirus-induced late antigens in women with imminent abortion in comparison to women with normal pregnancy]. PMID- 6295029 TI - A simple and inexpensive method for detection of BLV infected cattle based on a modified ELISA principle. PMID- 6295030 TI - Early diagnosis of virus induced bovine leukosis in milk by a simple modified ELISA test. PMID- 6295032 TI - [Elaboration of a method of stabilizing the protective fraction of pertussis microbes]. AB - A method for stabilizing adsorbed preparations of the protective fraction of B. pertussis has been developed; according to this method, a colloid suspension of protective protein in phosphate buffer is obtained, the protein is then adsorbed on aluminium hydroxide gel and lyophilized with 10% of sucrose. If stored at 4 degrees C, these dried preparations have been found to retain their immunogenicity for 1 year. PMID- 6295031 TI - [Current methods of detecting the hepatitis A virus and its antibodies]. PMID- 6295033 TI - [Isolation of a purified theta-hemolysin from C1. perfingens type A and its use for titrating toxins and sera]. AB - Purified Cl. perfringens theta-hemolysin with a specific activity of 15-16 X 10(5) HU per mg of protein nitrogen, toxic for mice when injected intravenously, has been obtained by precipitation with zinc chloride, gel filtration through Sephadex G-100 and ion-exchange chromatography on DEAE Sephadex A-50. 1 Dcl of the toxin is 0.2-0.4 mg. The yield of theta-hemolysin amounting to 20-25% has been achieved, the degree of its purification reaching 80- to 120-fold. Equine anti-theta hemolytic serum has been obtained. The method for the determination of the anti-theta toxin-binding properties of Cl. perfringens toxins and the quantitative content of anti-theta-hemolysin in sera has been developed. PMID- 6295034 TI - [Changes in the cAMP level in macrophages in Salmonella typhimurium phagocytosis]. AB - The effect of a virulent S. typhimurium strain and its cAMP-deficients mutant on the level of cAMP in macrophages in the process of phagocytosis has been studied. The virulent strain has been shown to induce the 3-fold increase of the level of cAMP in macrophages, while the mutant renders no such effect. PMID- 6295035 TI - [Genetic transfer of pAP38, pAP39, and pAP41 plasmids into E. coli, Erwinia, and Hafnia strains]. AB - Genetic transfer factors pAP38, pAP39 and pAP41 can be transferred to E. coli, both typed and untypecd, as well as to Erwinia and Hafnia strains, with different frequency (10(-2) to 10(-8)). The transconjugates thus obtained possess donor activity and can transfer the factors they have received to recipient E. coli strains. After transfer these factors are stably maintained in a new host for at least 10 days. Under the action of ethidium bromide or elevated temperature the elimination of the transfer factors from host bacteria is observed. The studied transfer factors pAP38, pAP39 and pAP41 (F-like factors) are plasmids fi+. PMID- 6295036 TI - [Characterisation of major biological properties of Bordetella bacteriophages]. AB - The main biological properties (morphology of negative colonies, parameters of adsorption and single development cycle) of B. pertussis and B. bronchiseptica phages, isolated spontaneously and by induction with mitomycin C, were studied. To compare these characteristics, one B. parapertussis indicator strain was used, and the experiments were carried out under identical conditions. Highly active sera were obtained with the use of complete Freund's adjuvant. B. pertussis phages isolated from the strains of different serovars were serologically related, but not identical, and differed in their constant characterizing their rate of neutralization with homologous antisera. The adsorption of the phages on homologous strains was more intensive than on the cells of B. parapertussis indicator strain. However, the authors failed to observe the further development of the phages in the host cells. PMID- 6295038 TI - [Status of the electromechanical system of a muscle in myasthenia]. PMID- 6295037 TI - [Viral hepatitis with clinical and epidemiological manifestations similar to hepatitis A but of a different etiology]. AB - A case of viral hepatitis in man, appearing as the result of infection caused by pooled concentrated suspensions of fecal samples collected from patients having had repeated hepatitis infection during the period of 1-2 years, is described. Though this infection was similar to hepatitis A in many clinical and epidemiological signs, the possibility of its etiologic relationship with hepatitis virus A was positively excluded; there was also no evidence of the participation of hepatitis virus B in the process. Immunoelectron microscopy of excretions collected at the acute stage of the disease revealed the presence of spherical viral particles 27-30 nm in diameter. Antibodies capable of reacting with these particles were detected in the sera of patients having had 2 kinds of hepatitis and in the sera of patients having the 1 kind of hepatitis in the focus of infection where repeated cases of hepatitis had been observed. No such antibodies were found in the sera of patients with hepatitis A alone and in the set of standard sera specific to viruses causing hepatitis A and hepatitis non A, non B. The authors believe that 2 kinds of hepatitis with the fecal-oral mechanism of transmission exist and propose to name their causative agents hepatitis viruses A, type 1 and type 2. PMID- 6295039 TI - [Prognostic findings in histopathology in bronchopulmonary carcinoma after surgery]. AB - The prognostic value of hystological cell type, nuclear grading and mitotic index has been examined in 48 carcinomas of the lung after radical surgery. Three hystological cell types have been identified: squamous cell carcinoma (48%), large cell anaplastic carcinoma (33%) and adenocarcinoma (19%). The Nuclear Grade, ranging from 0 to 4, has been classified as low (N.G. less than 2) and high (N.G. greater than or equal to 2). The Mitotic Index (M.I.) has been considered as the mean of the number of the mitosis observed in 10 microscopic high power fields. (400 X). The 5 year survival rate is significantly higher in patients having large cell carcinoma or squamous cell carcinoma in relation to N.G. less than 2 and M.I. less than mean. PMID- 6295040 TI - [Local cellular immune response in bronchopulmonary carcinoma after surgery in relation to long-term prognosis]. AB - The prognostic value of local stromal cell reaction has been examined in 48 carcinomas of the lung (23 squamous cell carcinomas, 16 large cell anaplastic carcinomas, 9 adenocarcinomas) after radical surgery. The intensity of the peri and intratumoral lymph cell (R.L.), plasma cell (P.R.) and macrophage (M.R.) reactions has been classified in low, moderate and high reactions. The 5 year survival rate is significantly higher in patients having large cell carcinoma and squamous cell carcinoma in relation to a high or moderate lymphocytic reaction. There is no correlation between the age of the patient and the stromal cell reaction. PMID- 6295041 TI - Effect of phthalate ester metabolites on rat liver. PMID- 6295042 TI - Decreasing incidence of acute appendicitis, with special reference to the consumption of dietary fiber. AB - The incidence of acute appendicitis and the total number of appendectomies performed in the adult population admitted to our medical center are both decreasing. The cause is not clear. Better nutrition and the wide-spread use of antibiotics are two possible factors which come to mind, but we know of no scientific evidence that these are responsible. There has been a slight decrease in the mean dietary fiber content in the Swedish diet during the last three decades. The decreased incidence of acute appendicitis cannot therefore be correlated with an increase in the mean dietary fiber intake. PMID- 6295043 TI - Guar and gastric emptying in non-insulin dependent diabetes. AB - A gamma camera was used to measure gastric emptying in 10 non-insulin dependent diabetics and 10 control subjects and after a breakfast meal of porridge labelled with 113mIn. In the diabetics there was a more rapid early phase and a more prolonged later phase of gastric emptying compared with controls. Incorporation of 10 g guar with the meal prolonged the later phase of gastric emptying in controls. However, guar did not significantly alter gastric emptying in the diabetics, although postprandial plasma glucose concentrations were reduced. The study demonstrates abnormalities of both the early and later phases of gastric emptying in an unselected group of non-insulin dependent diabetics. Guar reduced plasma glucose concentrations without affecting gastric emptying in these patients. PMID- 6295045 TI - Some circuit operations in the mammalian brain. AB - There is an account of the basis neuronal connectivities of the spinal cord with the Sherringtonian principles of divergence and convergence. Neurones act synaptically either as excitatory or as inhibitory, depending on the specific transmitter substances liberated. Inhibitory neurones usually act either in a feedback or a feedforward manner. Voluntary movement is considered in relation to the instructions delivered to the motor cortex in order to produce the discharges down the pyramidal tract that evoke the required movement. There is an account of the three lines of evidence which indicate that in voluntary movements the primary neural event arises in discharges of neurones of the supplementary motor area (SMA). There are three main circuits from the SMA that activate subroutines concerned in the preprogramming of movements: (1) SMA to the basal ganglia, thence to the thalamus with a collateral line through the substantia nigra, thence to the association cortex; (2) SMA to cerebellar hemisphere via the pontine nuclei, thence to the nucleus dentatus, to the thalamus, to the association cortex, and (3) SMA to association cortex both frontal and parietal. According to the SMA hypothesis the liaison brain for intention is located in the SMA, there being reciprocity of informational flow from the mental events of intention to the neuronal events in the SMA. PMID- 6295044 TI - To what extent does the artificial pancreas facilitate the surgery of preoperatively not localized insulinomas? AB - The patient study has been performed in order to evaluate the usefulness of the artificial pancreas in the surgical management of previously not localized insulinomas. In the 4 patients studied, blood glucose was maintained both overnight and during surgery up to a preselected individualized level in order to avoid hypoglycemia. During surgery, only one patient required dextrose infusion. The continuous intrasurgical monitoring of blood glucose in the 4 cases examined showed that: (1) anesthesia induction, surgical incision and viscera mobilization were accompanied by a rise in blood glucose (10.30 mg/dl), reaching the highest levels 30-40 min after the start of the operation; (2) adenoma manipulation was followed by a drop in blood glucose (10.40 mg/dl), reaching the lowest level after 30-40 min; (3) adenoma resection was followed by a rise in blood glucose (25-40 mg/dl), particularly evident after 30-40 min. It is concluded that the artificial pancreas is certainly useful during surgery of insulin-producing tumors, allowing continuous monitoring of glycemia and avoiding dangerous blood glucose excursions: however, when the insulinoma is not identified during surgery, the periods elapsing between the surgical phases and the blood glucose changes observed can be too prolonged to ensure successful conservative serial pancreatectomy in all cases. PMID- 6295046 TI - [Heteromorphic glioblastoma. Analysis of 77 cases]. PMID- 6295047 TI - Experimental allergic encephalomyelitis: modification of optic nerve pathology by antecedent virus infection. PMID- 6295049 TI - Sarcoidosis of the peripheral nerve: a histological and ultrastructural study of two cases. PMID- 6295048 TI - Intranuclear inclusions in the neurons of senescent rats. PMID- 6295050 TI - The pathology of a sensory neuropathy affecting Long Haired Dachshund dogs. PMID- 6295052 TI - The temporary threshold shift (TTS) and its relationships to the temporal characteristics of the acoustic stimulus. PMID- 6295051 TI - Effect of glycerol on the EP decrease caused by furosemide. AB - A change in endocochlear potential (EP) following furosemide injection was observed in regard to prior glycerol administration in two groups of guinea pigs. In one group, various doses of furosemide (20, 30, 40, 50 mg/kg) were injected, while in the other, glycerol (50 v/v %, 1 ml/kg) was injected prior to the furosemide (20, 30, 40 mg/kg) injection. In the glycerol-furosemide group, the decrease in EP was 40% greater than in the furosemide group. Therefore, glycerol was thought to potentiate the EP lowering action of furosemide. Such an effect of glycerol was assumed to be resulted by facilitating the access of furosemide to the site of action in the stria vascularis. PMID- 6295053 TI - Brain stem potential and its relationships to the temporal characteristics of the acoustic stimulus. PMID- 6295054 TI - Blood parameters, heart activity and their relationships to the temporal characteristics of the acoustic stimulus. PMID- 6295055 TI - Characteristics of the acoustic reflex elicited by pure tones and white noise bursts with different overall acoustic energy. PMID- 6295056 TI - Relationships between ambient acoustic energy and inner ear. Introduction, bibliographical survey and general remarks. PMID- 6295057 TI - Epidemiology of acute otitis media in children. PMID- 6295058 TI - Ultrasonic examination of the paranasal sinuses. PMID- 6295059 TI - Vasoactive intestinal polypeptide (VIP) in children with neural crest tumours. AB - The concentration of vasoactive intestinal polypeptide (VIP) in plasma was measured in 22 children with neural crest tumours (NCT) during a 5-year period; the mean concentration of VIP in plasma was 22.5 pmol/l (interval 2.0-95.0 pmol/l). To establish a reference interval the plasma concentration of VIP was measured in 41 children without tumours; the mean concentration of VIP in plasma was 6.2 pmol/l (interval 0.5-19.0 pmol/l). Of the 22 children with NCT 16 had a plasma concentration of VIP within the normal range while 6 children (27%) had elevated plasma concentration of VIP between 28 and 95 pmol/l. Only one child, whose plasma concentration of VIP was 95 pmol/l had diarrhoea. Elevated plasma concentration of VIP in children with NCT but no diarrhoea has not previously been described. The urinary excretion of vanillylmandelic acid (VMA) was increased in 18 of the children with NCT (82%). In 2 of the children with normal excretion of VMA the concentration of VIP in plasma was elevated. Thus, the plasma concentration of VIP may be a supplement to VMA as a tumour marker in some cases of NCT. PMID- 6295060 TI - Childhood acromegaly successfully treated with interstitial irradiation using yttrium-90. AB - A child with a growth hormone producing tumour presented at the age of 4 1/2 years. The onset of the disease was at 18 months of age. Treatment was given with three doses of interstitial irradiation using yttrium-90 implants. There were no local complications from the procedures. Now, 11 years after diagnosis, she is asymptomatic, of normal appearance, and her height and the size of the pituitary fossa are normal. Growth hormone levels are almost normal, thyroid function is intact, and she is maintained on prednisone and sex hormones. PMID- 6295061 TI - Induction and abrogation of suppressor cell function in humans: effect on B cell activation by different polyclonal activators. AB - In a reverse plaque forming cell (PFC) assay, the effect of concanavalin A (Con A)-induced suppressor cells on polyclonally activated B lymphocytes was studied. Differentiation to PFC, as induced by pokeweed mitogen (PWM), Staphylococcus aureus or Epstein-Barr virus (EBV) was in all cases suppressed by the addition of ConA-pretreated cells. The EBV-stimulated cells showed the smallest PFC response and appeared least amenable to suppression. IgA-secreting cells were less suppressible than IgM- and IgG-secreting cells regardless of the polyclonal B lymphocyte activator used. The suppressor cells were radiosensitive. PMID- 6295062 TI - [Applications of A. C. oscillopolarographic titration in pharmaceutical analysis. II. Titration of disodium dimercaptosuccinate with Cd]. PMID- 6295063 TI - Interaction of hydroflumethiazide and 2,4-disulfamyl-5-trifluoromethylaniline with cyclic AMP phosphodiesterase and carbonic anhydrase. AB - The inhibitory effect of hydroflumethiazide (HFT) and its metabolite, 2,4 disulfamyl-5-trifluoromethylaniline (DTA) on cyclic AMP phosphodiesterase and the binding of HFT and DTA to carbonic anhydrase was studied in vitro. Significant inhibition of rat kidney low-Km cyclic AMP phosphodiesterase was observed with DTA concentration above 2.5 X 10(-4) mol/l and with HFT concentration above 1 X 10(-4) mol/l. 50% inhibition was observed at a DTA concentration of 1 X 10(-3) mol/l. Binding of DTA and HFT to commercially obtained bovine erythrocyte carbonic anhydrase was demonstrated by equilibrium dialysis. Data were consistent with one class of binding sites. The product of n (number of binding sites) and Kass (association constant) was 5 X 10(5) M for DTA and 3.3 X 10(4) M for HFT at 2 degrees. In human blood in vitro at 37 degrees, the equilibrium erythrocyte/plasma concentration ratio was 18 for DTA and 1.6 for HFT. It is concluded that HFT and DTA have approximately the same potency as cyclic AMP phosphodiesterase inhibitors, whereas DTA is more extensively bound by erythrocyte carbonic anhydrase. PMID- 6295064 TI - Binding of beta-carbolines and caffeine on benzodiazepine receptors: correlations to convulsions and tremor. AB - Compounds from both the beta-carboline (BC) and xanthine groups have been suggested to be the natural ligands for benzodiazepine (BZ) receptors. In this study we examined the effects of several BC's and caffeine, 1,3,7 trimethylxanthine, on the binding of 3H-flunitrazepam (3H-FZ) and beta-3H carboline-3-carboxylic acid ethyl ester (3H-BCCE) to the BZ receptors of rat and mouse brain. In mice, convulsion-producing doses of caffeine (120 mg/kg intravenously) and harmane (30 mg/kg intravenously) lowered the specific binding of 3H-FZ in vivo by 12-31%. A tremorogenic dose of harmaline (30 mg/kg intravenously) increased binding by 31%. Caffeine and harmane also slightly decreased the in vivo binding of 3H-BCCE, a compound that binds preferentially to the cerebellar type of BZ receptors. Harmaline stimulated the binding of 3H-BCCE only in the forebrain. Both harmaline and harmane increased by 41-111% the amount of 3H-BCCE that was distributed to the brain. In vitro BC's and caffeine displaced 3H-FZ from receptors in the rat brain with various Ki values (4.7 to 206.9 microM). The antagonism for BZ binding was competitive and in Scatchard analysis produced linear plots. Exceptions were harmaline and caffeine in the forebrain: both exhibited curvilinear plots for 3H-FZ binding. Harmaline increased the binding, and caffeine decreased it by altering the affinity of a subgroup of BZ receptors. In the hindbrain both harmaline and caffeine inhibited binding and produced linear plots. BC-induced tremor and convulsions unveil a large number of spare receptors in the brain, and these seem to be of the cerebellar type of BZ receptors. In addition, our results show that tremorogenic and convulsive BC's act differently on BZ receptors: during harmaline-induced tremor the affinity of some BZ receptors is increased, while harmane-induced convulsions are connected to direct occupation of BZ receptors. PMID- 6295065 TI - Reversal of enterotoxic diarrhoea by anaesthetic and membrane-stabilizing agents. AB - The effect of membrane-stabilizing and sedative drugs on enterotoxic diarrhoea was studied in mice. Fluid secretion was induced in ligated loops of the small intestine by challenge with cholera toxin (CT), heat-labile enterotoxin (LT) from Escherichia coli or dibuturyl-cyclic AMP (dB-cAMP). Chlorpromazine, melperone, diazepam, mebumal, ketamine and ethanol all inhibited CT-induced hypersecretion. ED50 being 1.5, 4, 4, 35, 70 and 1500 mg/kg, respectively. The drugs also blocked CT-stimulation of adenylate cyclase, which mediates the action of CT. Concomitant with the antisecretory effect a sedative effect was induced, as judged by the motility and righting reflex of the animals. Hypersecretion by E. coli and LT was also totally blocked by chlorpromazine, diazepam, ketamine and ethanol. In contrast, the secretion by dB-cAMP which bypasses adenylate cyclase in its action, was not affected by diazepam and was only partly reversed by chlorpromazine, ketamine and ethanol. The reversal of dB-cAMP-secretion is probably due to enhanced absorption, rather than inhibition of adenylate-cyclase. The use of membrane-stabilizing drugs may represent a new principle for pharmacological treatment of diarrhoea. PMID- 6295066 TI - Comparison of the effects of different arachidonic acid metabolites on cyclic nucleotide accumulation in human peripheral lymphocytes. AB - The effect of PGE1, PGE2, PGD2, PGF2 alpha, PGI2, PGG2, PGA1, 12L-HETE, arachidonic acid, 15- HPETEa and linolenic acid on the accumulation of cyclic AMP in human peripheral lymphocytes was studied. PGE1, PGE2 and PGD2 were essentially equipotent as stimulators of cyclic AMP accumulation (threshold at about 10(-8)M and EC50 about 0.15 microM), PGF2 alpha was about 20 times less potent, while PGG2, 12L-HETE, 15-HPETE, PGA1 and linolenic acid were inactive. PGI2 caused a weak stimulation between 5 and 600 nM and a secondary stimulation above 3 microM. Arachidonic acid had no effect on cyclic AMP levels up to 100 microM. PGE1, PGD2, PGI2 and PGF2 alpha increased cyclic GMP in the concentrations that produced a rise in cyclic AMP, but the cyclic GMP increase was of smaller magnitude. Exogenous arachidonic acid was converted mainly to 12L-HETE, HHT and thromboxane B2 by lymphocyte suspensions. This conversion could be accounted for by contamination with blood platelets. The results show that the degree of cyclic AMP accumulation in human lymphocytes following stimulation of arachidonic acid metabolism will be critically dependent upon which prostaglandins are in fact formed by cells surrounding the lymphocytes. PMID- 6295067 TI - Relaxant beta-receptors in the trachea, but not prejunctional alpha 2-receptors in the tracheal cholinergic neurones, are subjected to homologous desensitization. AB - Guinea-pigs were pretreated with either isoprenaline, terbutaline or the alpha 2 agonist B-HT 920 in order to asses the hypothesis that beta-and alpha 2 receptors in trachea are subjected to homologous desensitization. In these experiments the beta receptor activity was investigated on tracheal ring preparations contracted with carbacholine. Both in the isoprenaline and the terbutaline pretreated group the relaxant responses to beta agonists were diminished. Pretreatment with B-HT 920 did not affect the sensitivity of the trachea to beta stimulation. In order to asses the responsiveness of alpha 2 receptors the trachea was contracted by electrical field stimulation in the presence of propranolol. During these conditions contractions were mediated by activation of cholinergic neurones and inhibitory effects of alpha stimulation were due to inhibition of the cholinergic neurotransmission by stimulation of prejunctional alpha 2 receptors. In these tests neither isoprenaline, terbutaline nor B-HT 920 pretreatment affected the responsiveness of alpha stimulation to inhibit the electrically induced contractions. PMID- 6295068 TI - Anoxia increases potassium conductance in hippocampal nerve cells. AB - The effect of anoxia on nerve cell function was studied by intra- and extracellular microelectrode recordings from the CA1 and CA3 region in guinea pig hippocampal slices. Hyperpolarization and concomitant reduction of the nerve cell input resistance was observed early during anoxia. During this period the spontaneous activity first disappeared, then the evoked activity gradually disappeared. The hyperpolarization was followed by depolarization and an absence of a measurable input resistance. All the induced changes were reversed when the slice was reoxygenated. Reversal of the electro-chemical gradient for Cl- across the nerve cell membrane did not affect the course of events during anoxia. Aminopyridines blocked the anoxic hyperpolarization and attenuated the decrease of membrane resistance, but had no effect on the later depolarization. Blockers of synaptic transmission. Mn++, Mg++ and of Na+-channels (TTX) were without effect on the nerve cell changes during anoxia. It is suggested that the reduction of nerve cell excitability in anoxia is primarily due to increased K+ conductance. Thus, the nerve cells are hyperpolarized and the input resistance reduced, causing higher threshold and reduction of synaptic potentials. The mechanism of the K+-conductance activation is unknown at present. PMID- 6295069 TI - Intracisternal administration of avian pancreatic polypeptide lowers respiration rate and enhances the clonidine induced reduction of respiration rate in alpha chloralose anesthetized rats: possible interactions with an alpha 2-adrenergic receptor. PMID- 6295070 TI - Rat plasma and hypothalamic beta-endorphin levels fluctuate concomitantly with plasma corticosteroids during the day. PMID- 6295071 TI - Increased excretion of urinary cyclic GMP in primary hepatoma and preneoplastic liver. AB - Urinary excretion of cyclic GMP (cGMP) and the plasma level of cyclic AMP (cAMP) were determined in patients with liver diseases. The urinary excretion of cGMP, expressed on the basis of creatinine excreted per day, was at significantly higher levels not only in primary hepatoma but also in liver cirrhosis, while the plasma level of cAMP was higher only in liver cirrhosis. Thus, the ratio of urinary cGMP excretion to plasma cAMP level in primary hepatoma was significantly higher than that in liver cirrhosis. In cirrhotic patients studied by catheterization, the level of cGMP in the hepatic vein was significantly lower than that in the superior mesenteric or portal vein, indicating the uptake of cGMP by the liver. Since cGMP excretion correlated with KICG both in liver cirrhosis and primary hepatoma, the increased cGMP excretion appeared to be explained by a reduced uptake of cGMP by the liver. PMID- 6295073 TI - Receptive fields of neurons in the cats visual cortex after a change of alertness level. AB - Exact and reliable changes in receptive fields (RF) of neurons in the primary visual cortex were revealed after the action of extravisual stimuli such as air puff to the corner of a closed eye which leads to an enhancement of alertness level. Such stimulus evoked a change in the configuration and size of RFs in most investigated units. This lasted from 15 to 30 min and in most cases the size of RFs excitatory zones decreased, while the size of inhibitory surround increased. This effect could be seen at different levels of visual adaptation. Under light and short anesthesia (sombrevin) the main picture of RF change was different: increase of excitatory zones and nearly equal probability of the opposite changes in inhibitory RF parts lasting from 5 to 30 min. The role of intracortical inhibitory systems in the modulation of RF size and form is discussed. The possible behavioral meaning of the observed effects may consist in the fine description of signal properties in the small RFs at high alertness level, while under opposite conditions enlarged RFs can serve to improve signal detection and organization of orienting reaction even to a weak new stimulus. PMID- 6295072 TI - Electrophysiological analysis of conditioned taste aversion in rats. AB - The memory mechanisms of conditioned taste aversion (CTA) were examined using electrophysiological methods. Presentation of the aversive taste to CTA trained animals inhibited unit activity in gustatory cortex, amygdala and ventromedial hypothalamus and caused delayed excitation of neurons in lateral hypothalamus. Lick-triggered rewarding stimulation of medial forebrain bundle substituted the taste CS in CTA experiments. The same stimulation triggered by nose-poking failed to be associated with subsequent poisoning. Importance of specific brain areas for CTA retrieval was assessed by the effect of lick-triggered stimulation of the examined site on gustatory discrimination. The disruption threshold was lowest in amygdala and lateral hypothalamus. Stimulation of other brain structures did not interfere with gustatory discrimination at current intensities subthreshold for disruption of licking. Unilateral electrical stimulation of vestibular nuclei following ingestion of saccharin elicited marked aversion to this taste. PMID- 6295074 TI - A model for xenotransplantation of human malignant astrocytomas into the brain of normal adult rats. AB - Transplantation of human brain tumours into the brain of normal laboratory animals is still considered to be unsatisfactory by many researchers, despite the fact that the brain is considered an immunologically privileged site. We present in this paper a model of xenotransplantation fo human astrocytomas grade III-IV into the brain of normal, adult Sprague-Dawley rats with good take rates, i.e. takes in two thirds of the animals, half of these with large, infiltrating tumours. The transplants are placed using a microsurgical technique in the vessel rich choroidal fissure in the host brain from where rapid vascularization occurs. The technique has previously been used for CNS-regeneration studies. This model should provide an excellent opportunity to study human malignant astrocytomas in a milieu as natural as possible. PMID- 6295076 TI - The pituitary: cerebrospinal fluid barrier in the primary empty sella syndrome. PMID- 6295075 TI - S-100 protein in human glial tumours. Qualitative and quantitative studies. AB - The authors studied a total of 48 human glial tumours for S-100 protein, an extremely acidic, protein specific to the nervous system, by immunohistochemistry and by micro-complement fixation assay in order to evaluate S-100 protein as an index for malignancy of glial tumours. All of 48 glial tumours analyzed in the present study demonstrated variable amounts of S-100 protein which might serve as a biochemical cell marker for glial tumours. The mean value of S-100 protein content in 3 ependymomas is higher than those of 19 low-grade (grades I, II) astrocytomas and 26 high-grade (grades III, IV) astrocytomas, being lowest in the latter. A statistically significant (P less than 0.001) difference in S-100 protein levels between low- and high-grade astrocytomas is observed, but not for ependymoma. At present, however, no correlation can be found between S-100 protein content of a tumor and the patient's survival time. Immunoperoxidase staining for S-100 protein in high-grade astrocytomas is generally weak in intensity and heterogeneous throughout the section, whereas that in low-grade astrocytomas and ependymomas is relatively strong and homogeneous, indicating that high-grade astrocytomas consist of a more heterogeneous population of tumour cells in terms of S-100 protein. These results show that the investigation of S 100 protein in a glial tumor is valuable to a certain extent in assessing the degree of differentiation or malignancy of the tumour. PMID- 6295077 TI - Properties of follicle stimulating hormone binding inhibitors found in physiological fluids. PMID- 6295078 TI - Oocyte maturation inhibitor: a 1981 perspective. PMID- 6295079 TI - Role of cumulus cells in oocyte maturation. PMID- 6295080 TI - Actions of hormones and other factors upon oocyte maturation. AB - Oocyte maturation is controlled by a combination of hormonal and local follicular factors. Osmolarity, pH, and perhaps Ca2+ concentration of the surrounding medium are also important. Follicular fluid contains a low molecular weight OMI which acts to keep the oocyte from maturing. Luteinizing hormone added to cultured cumulus enclosed porcine oocytes can reverse the inhibitory action of OMI. The level of OMI in the follicular fluid appears to decrease as the follicle matures. Addition of FSH and prolactin to cultured granulosa cells stimulates OMI secretin whereas addition of testosterone or dihydrotesterone brings about a decrease in OMI secretion. Elevated LH in vivo may bring about oocyte maturation before ovulation by (a) an antagonist action on OMI; (b) stimulating the synthesis of testosterone by theca cells and thus inhibiting the synthesis of OMI by granulosa cells; and (c) action on the granulosa cells to promote luteinization which may also cause a decrease in OMI synthesis. The hastened oocyte maturation associated with follicular atresia could be due to a decline in OMI due to granulosa cell death and/or elevated follicular androgens. PMID- 6295081 TI - Intraovarian actions of GnRH. PMID- 6295082 TI - Regulation of ovarian granulosa and luteal cell functions by gonadotropin releasing hormone and its antagonist. PMID- 6295083 TI - LH-binding inhibitors from the corpus luteum. PMID- 6295085 TI - Regulation of cholesterol acquisition and utilization in the corpus luteum. AB - In this report we have focused on two aspects of control of corpus luteum function; the regulation of lipoprotein uptake and the modulation of cholesterol side chain cleavage through alterations in the phospholipid milieu. Until recently, a role of gonadotropins in the acquisition of lipoprotein-carried cholesterol had not been generally appreciated. However, it is likely that the regulation expression of cell surface lipoprotein receptors will prove to be an important function of tropic hormones, particularly LH, since luteal cells generate progestins at a substantial rate and must, therefore, have an equally substantial supply of cholesterol. Factors other than gonadotropins may have a function in control of lipoprotein receptors and it would not be surprising if luteolytic factors, such as prostaglandin F2 alpha, are found to diminish lipoprotein-uptake as part of their action in reducing steroidogenesis. The control of mitochondrial pregnenolone synthesis by gonadotropin-induced alterations in phospholipid composition of mitochondrial membranes represents a novel concept. Whether certain polar lipids will emerge with the status of intracellular messengers remains to be seen. In any event, our observations clearly demonstrate the possibility for such a mechanism of control. PMID- 6295084 TI - Characterization of LH/hCG receptor binding inhibitor in corpora lutea of human and sheep ovaries. PMID- 6295086 TI - Intraovarian regulation of granulosa-cell replication. PMID- 6295087 TI - Comparison of granulosa and sertoli cells at various stages of maturation: similarities and differences. PMID- 6295088 TI - Intracellular inorganic phosphate and ATP levels in human blood erythrocytes, leucocytes and platelets in normal subjects and in diseases associated with altered phosphate metabolism. PMID- 6295089 TI - Regulation of phosphate excretion by the diseased kidney: the intrinsic proximal tubular adaptation. PMID- 6295090 TI - Parathyroid hormone resistance in renal allograft recipients. PMID- 6295091 TI - Experimental alcoholism induces phosphorus and magnesium deficiency in skeletal muscle. PMID- 6295092 TI - Development and application of an exchange assay for 1,25-dihydroxyvitamin D3. PMID- 6295093 TI - Metabolic abnormalities underlying the pathogenesis of vitamin D resistant rickets (VDRR). PMID- 6295094 TI - Calcium as intracellular messenger in hormone action. PMID- 6295095 TI - Functional properties of parathyroid hormone receptors in kidney and bone. PMID- 6295096 TI - Effects of parathyroid hormone on glomerular ultrafiltration in the rat. PMID- 6295097 TI - 1,25-dihydroxycholecalciferol (calcitriol): effect on phosphate (PO4) absorption by isolated pars recta. PMID- 6295098 TI - Neuronal degenerations of aging. PMID- 6295099 TI - Nonfamilial amyotrophy with dementia or multisystem degeneration and other neurological disorders. PMID- 6295100 TI - The pathogenesis of motor neuron disease: perspectives from neurotoxicology. PMID- 6295101 TI - Persistent infection by poliovirus: experimental studies. PMID- 6295102 TI - Nucleic acid probes in the study of amyotrophic lateral sclerosis. PMID- 6295103 TI - Selective vulnerability of neural cells to viral infections. AB - A number of viruses selectively infect neurons and, in some cases, specific populations of neurons. The susceptible neuron need not be permissively infected to cause acute or chronic disease; therefore, infectious virus may not be recoverable and morphologically identifiable viral structures may not be detectable by ultrastructural structures. Polioviruses and the neurotropic murine retrovirus both cause paralytic disease with major pathological changes in motor neurons of the spinal cord. Both produce disease more readily in later life; in poliovirus because the mature animals are more susceptible to acute infection, and in the neurotropic retrovirus infections because of the long incubation period of the natural infection. In the acute inflammatory poliovirus infections, the motor neurons appear to be selectively infected and lysed by the virus, whereas in the chronic noninflammatory retrovirus infection, the effect may be indirect or may result from nonpermissive infection. PMID- 6295104 TI - Aspects of the ultrastructure of amyotrophic lateral sclerosis. PMID- 6295105 TI - Slow virus infections of ovine lung. PMID- 6295106 TI - Anti-inflammatory effect of LA 2851 and reference drugs on some models of inflammation. Investigation of the mechanism of action. AB - LA 2851 (2-4-diamino-7-methyl-pyrazolo (1,5-a) 1,3,5-triazine), a bronchodilator and antiallergic compound, in type I hypersensitivity, has been tested orally for activity against carrageenan oedema and complement dependent, reverse passive Arthus (RPA) and zymosan oedema in rats. Pharmacokinetic determinations were also realized in order to correlate plasma blood levels and pharmacological activity. LA 2851 was found active in the first test but showed a more marked effect in the immunologically mediated RPA reaction and zymosan oedema. Among reference drugs tested, theophylline showed the same pattern in contrast with non-steroidal anti inflammatory agents. LA 2851 and theophylline, using a superfused lung preparation, were found ineffective on the synthesis of cyclooxygenase products from arachidonic acid. LA 2851 as theophylline inhibited cAMP phosphodiesterase (PDE) but this inhibition does not seem to be involved in their anti-inflammatory activity since papaverine, a potent inhibitor of PDE, was totally inactive. The activity on RPA and zymosan inflammation was achieved at the drug plasma level in the range of those required to relax the trachea. The same antagonism was obtained with the 2 drugs at a lower plasma level with LA 2851 than with theophylline for the same dose administered (25 mg/kg). LA 2851 and theophylline did not inhibit all the components of the inflammatory process since maximum inhibition did not exceed 60% up to 200 mg/kg and 100 mg/kg respectively. PMID- 6295107 TI - Prostaglandin-mediated inhibition of lymphokine secretion in normal individuals and patients with progressive systemic sclerosis (scleroderma, PSS). AB - The sensitivity of peripheral blood lymphocytes to E-type prostaglandin-mediated inhibition of lymphokine secretion was examined in 3 groups of individuals; normal controls, hospitalized patients, and patients with progressive systemic sclerosis (PSS, scleroderma). Leukocytes were stimulated by a polyclonal T-cell activator, phytohemagglutinin, and the release of the lymphokine, leukocyte migration inhibitory factor (LIF), was measured in the presence or absence of exogenous PGE2 using a direct agarose droplet migration inhibition technique. Leukocytes of scleroderma patients were found to be hyporesponsive to E-type prostaglandin (i.e., lymphokine secretion by these cells was not inhibited at concentrations of PGE2 between 2.8 X 10(-8) and 2.8 X 10(-5) M). In addition, a marked sex difference in PGE responsiveness was found to exist among normal controls, whereby females were hyporesponsive during the latter half of the menstrual cycle. It is possible that this deficit may facilitate, in part, the development of connective tissue diseases in women of childbearing age. The inability to suppress lymphokine production and arrest persistent immune reactivity, coupled with the known ability of lymphokines to augment fibroblast collagen production, offers a a reasonable explanation for the accumulation of tissue collagen in scleroderma. PMID- 6295108 TI - Lysosomal enzymes and metabolic activity of polymorphonuclear leukocytes from patients with systemic lupus erythematosus and from experimental animals after levamisole treatment. AB - The activity of beta-D-glucuronidase (BDG) was lowered and the activity of myeloperoxidase (MPO) was elevated in polymorphonuclear leukocytes (PMNs) of SLE patients compared with normal subjects. After levamisole treatment, the activities of BDG, MPO, and lysozyme rose in PMNs of SLE patients. A higher activity of lysozyme was also observed in peritoneal PMNs of rabbits after levamisole administration. During incubation of human phagocytizing and non phagocytizing PMNs, no effects of levamisole at concentrations of 10(-3) to 10( 5) M were observed on the release of lysosomal enzymes. These concentrations of levamisole also did not influence INT reductase activity and production of superoxide by normal human PMNs in the presence of zymosan particles. These findings suggest that levamisole might have an effect on the actual level of lysosomal enzymes in PMNs rather than on their release. PMID- 6295109 TI - A comparison of superoxide production by human eosinophils and neutrophils. AB - Previous studies have demonstrated that human eosinophils have a more active hexose monophosphate shunt than neutrophils. However, measurements of superoxide anion production have been variable. In this study human eosinophils are shown to produce one and a half times as much superoxide as neutrophils during a 2-h incubation. The rates of production differed only after the first 30 min of incubation. The production was inhibited by superoxide dismutase in a dose responsive manner. PMID- 6295110 TI - Effects of nonsteroidal anti-inflammatory drugs on rat gastric mucosal phosphodiesterase activity. AB - In the present study, the effects of acetylsalicylic acid, diclofenac, ibuprofen, indomethacin, naproxen, phenylbutazone, proquazone, fluproquazone (RF 46-790 N), sulindac (sulfoxide and sulfide forms) and tolfenamic acid were compared on rat gastric mucosal cyclic nucleotide phosphodiesterases (PDEs). Some of the drugs inhibited PDEs effectively, the Ki values being clearly lower than those of theophylline. Mostly the type of inhibition was apparently competitive. Acetylsalicylic acid and ibuprofen were ineffective. No unambiguous correlation between the inhibition of mucosal PDEs and clinically observed gastric irritation was found. However, the inhibition of PDEs may modulate gastric side-effects of NSAIDs. PMID- 6295112 TI - Oxygen radicals. AB - Superoxide radicals play a role in at least three different levels of the biochemistry of inflammation, 1) They are produced sometimes excessively by macrophages and polymorphonuclear neutrophils during phagocytosis (bacterial killing). 2) A plasmatic neutrophil chemotactic factor is activated by O2 -. and 3) These radicals can cause chromosome breakage and other aberrations and also activate a plasmatic clastogenic factor present in various autoimmune diseases and in high energy irradiation cases. Superoxide dismutases can probably be useful for the control and treatment of autosustaining inflammatory conditions. Pharmacological application of superoxide dismutase is governed by a number of parameters including 1) Circulation time, 2) Penetration of cells, 3) Organ specificity, 4) Intracellular localization of the exogenous superoxide dismutase. Some recent experimental results in these areas will be described using both free superoxide dismutase and liposomal encapsulated enzyme. Although superoxide anions are the primary oxygen radicals produced in biological systems, they can also give rise to a cascade of other radicals such as hydroxy, carbonate and lipoperoxy radicals. Other enzymic systems for protection against uncontrolled oxidative processes exist and may have a future medical utility in certain cases. PMID- 6295111 TI - Interactions in connective tissues involving monocyte/macrophages and control of production of proteinases and proteinase inhibitors. AB - Using human articular chondrocytes in monolayer culture as an experimental system, we have been studying mechanisms of control of production and activity of neutral proteases which degrade connective tissue matrices. Soluble factors from cultured human blood mononuclear cells (MCF) or synovial fragment cultures (SF) stimulate the production of collagenase and proteoglycanase by chondrocytes. Chondrocytes also release a collagenase inhibitor (mol. wt. 26-31,000), which is similar to the tissue inhibitor of metalloproteinases (TIMP) synthesized by cultured mammalian tissues and this is reduced in cultures exposed to MCF or SF. Retinol and dexamethasone partially inhibit the factor-stimulated collagenase, but increase the amount of inhibitor, restoring it to control levels in the presence of MCF or SF. The effects of these agents in cellular interactions in vitro will be discussed in relation to their possible roles in the control of connective tissue turnover in vivo. PMID- 6295113 TI - Leukotriene B4: an inflammatory mediator with vascular actions in vivo. AB - Leukotriene B4 (LTB4) is a 5, 12-dihydroxy derivative of arachidonic acid generated by a variety of inflammatory cells via the lipoxygenase enzyme system. In vitro leukotriene B4 is a potent chemotactic and aggregatory agent; enhances neutrophil complement receptors; stimulates membrane calcium changes and causes contraction of lung parenchyma. In vivo LTB4 is a potent stimulator of vascular permeability in rats, rabbits, guinea-pigs and man particularly in the presence of a vasodilator such as PGE2. LTB4 causes a profound transient neutropenia and massive accumulation of neutrophils when injected into the dermis (rabbit and man), skin chambers (rabbit and man) or body cavities (guinea-pig, rat). PMID- 6295114 TI - Oxygen free radicals and leukotriene B4 induced increase in vascular leakage is mediated by polymorphonuclear leukocytes. AB - Topical application of the arachidonic acid metabolite leukotriene B4 (LTB4), known to be a potent chemotactic and chemokinetic substance or the superoxide radical producing enzyme xanthine oxidase (XO) initiate an increase in adhering polymorphonuclear leukocytes (PMN) along the venular endothelium and an increase in vascular leakage of fluorescein labelled dextran (Mw 150.000) in the hamster cheek pouch microvasculature. We have studied the contribution of PMN in the permeability response induced with LTB4 or XO. Animals made neutropenic by pretreatment with antineutrophil serum (ANS) obtained from immunized rabbits, showed a dramatically decreased leakage response to LTB4 or XO, whereas the response to histamine was unaffected. These results suggest that LTB4 and XO induced increase in vascular leakage is mediated by PMN adhering to the venular endothelium. Extensions can be made to acute inflammatory responses characterized by accumulation of PMN, vascular leakage and oedema formation. PMID- 6295115 TI - [Ca2+-dependent thiol protease (calpain) in cultured retinal pigment epithelial cells]. PMID- 6295116 TI - [Phagocytic activity of cultured retinal pigment epithelial cells: inhibition by melatonin and cyclic AMP, and reversion by taurine and cyclic GMP]. PMID- 6295117 TI - [The DNA restriction enzyme analyses of herpes simplex virus type 1 isolates from cases of ocular herpes]. PMID- 6295119 TI - Tail signs. PMID- 6295118 TI - Clostridium perfringens type A toxins in relation to nephritis and uric acid diathesis in captive willow ptarmigan (Lagopus l. lagopus). PMID- 6295120 TI - Nodule size and tail signs. PMID- 6295121 TI - Limited value of CT brain scans in the staging of small cell lung cancer. AB - Computed tomography of the brain was performed as part of the initial staging evaluation of 84 patients with small cell lung cancer. Brain scans indicative of metastatic disease were obtained in 12 (14%) patients, two of whom had no neurologic signs or symptoms. One of these had no other extrathoracic disease. Brain scans without evidence of metastatic disease were obtained in 72 patients, 58 (80.5%) of whom had no signs or symptoms suggestive of metastatic intracranial disease. In the 14 patients with neurologic symptoms but negative computed tomographic scans, other explanations than brain metastases were found. It was concluded that head scanning is a sensitive and accurate method of detecting central nervous system metastases in patients with small cell lung cancer. However, head computed tomography should not be included as part of the initial staging evaluation of the neurologically asymptomatic patients. In only one of 60 such patients did the brain scan change the initial clinical staging, which included chest films, liver and bone scans, and bone marrow biopsy. PMID- 6295122 TI - Primary linitis plastica of the rectum. PMID- 6295123 TI - Spectrum of sonographic findings in 125 renal masses other than benign simple cyst. AB - The sonographic findings of 125 renal masses that did not meet the sonographic criteria of benign simple cyst were retrospectively reviewed and correlated with the pathologic diagnosis. Of the 125 masses, 102 were renal cell carcinoma. These carcinomas, two of nine angiomyolipomas, and the 14 other renal masses of various histologic types exhibited a broad spectrum of sonographic findings. Seven of nine angiomyolipomas and the one multiloculated cystic nephroma had a rather characteristic sonographic appearance. With the knowledge of this spectrum of sonographic findings and strict attention to scanning techniques and sonographic criteria, the radiologist should be able to define the characteristics of a variety of renal masses and suggest the correct diagnosis. It should be possible to make the diagnosis of angiomyolipoma confidently if the characteristic sonographic and computed tomographic appearance of these tumors can be demonstrated. In some cases, the correct histologic diagnosis of a renal mass can be made only with biopsy or surgery. PMID- 6295124 TI - Bronchogenic carcinoma in breast carcinoma patients. AB - Forty-eight female patients who developed bronchogenic carcinoma synchronously with or after the occurrence of breast carcinoma are reported. The object of the study was to review the radiographic findings for possible clues to the diagnosis of bronchogenic carcinoma and to ascertain the histologic diagnoses observed. In 40 patients on whom films were available, the original radiographs before surgery for bronchogenic carcinoma were examined. The clinical records of the 48 patients were also reviewed. Histologic diagnosis of primary bronchogenic carcinoma was required. A solitary pulmonary lesion in a patient with breast carcinoma, a cavitated lesion, or a lesion that failed to respond to appropriate therapy for metastatic breast carcinoma suggested the diagnosis of bronchogenic carcinoma. An increase in the frequency of small cell carcinoma and a decrease in the frequency of adenocarcinoma was seen in bronchogenic carcinomas developing 5 years or more after radiation therapy for breast carcinoma. It was concluded that the benefits of radiation therapy in selected patients far outweighed the risk of bronchogenic carcinoma. PMID- 6295125 TI - Dyes, preservatives and salicylates in the induction of food intolerance and/or hypersensitivity in children. AB - We present 25 patients, aged between 18 and 153 months, with clinical symptoms suggestive of allergy to food antigens. After undergoing exhaustive studies (including case histories, cutaneous tests for reactions to food antigen, peripheral eosinophils, secretory and humoral immunity, determination of total IgE and of specific RAST, exclusion-provocation diets) and without being able to identify an offender, patients were submitted to oral provocation with different food additives (tartrazine, sunset yellow FCF, new coccine, erythrosine, sodium benzoate, 4-methyl hydroxybenzoate and acetylsalicylic acid) after 48 hours of exclusion from their diets of dyes, preservatives and salicylates. The results obtained reflect, at a global level, 57.89% of positivities for dyes, 34.21% for the benzoates and 7.81% for acetyl-salicylic acid. The low incidence of crossed intolerance phenomena should be emphasized (32% of the patients). The disparity of our results with those of other authors could be due to the age of our patients, the clinical patterns they present and the dietary habits of different countries and regions. PMID- 6295126 TI - A study of the circadian rhythm in 3, 5 cAMP in bronchial asthma after administration of ketotifen. AB - A study of the circadian rhythm of cAMP is carried out in ten patients suffering from mixed bronchial asthma, with a marked bacterial component and with sensitizations due to pneumo-allergens (house dust and Dermatophagoides). Cyclic AMP levels were determined before and after treatment with Ketotifen (HC-20511). At the same time, cAMP levels were determined in a control group of 10 healthy subjects in order to realize a comparative study with the asthmatic group. Blood was drawn beginning at 5 AM. at four hour intervals for 24 hours. During this period, medication was withdrawn from these patients, although in some cases antibiotics and mucolytics were administered later; these same patients were treated with Ketotifen in a dose of 1 mg every 12 hours for 5 days. In the blood samples, cAMP levels were determined through a method based on Gilman's technique (comparative protein binding) using the Cyclic AMP Assay Kit (The Radiochemical Center, England). Among the results obtained, there is a clear elevation of cAMP levels in all the determinations after the administration of Ketotifen, maintaining the circadian rhythm with a maximum peak in cAMP levels 5 hours after the drug was given. Thus an increase in all values when compared with basal levels were found arguing in favour of a protective action of this substance carried out through stabilization of the mast cell membrane and thus impeding the release of histamine. PMID- 6295127 TI - Prediction of subclinical perhexiline neuropathy in a patient with inborn error of debrisoquine hydroxylation. PMID- 6295128 TI - Left ventricular performance during exercise in patients with left bundle branch block: evaluation by gated radionuclide ventriculography. AB - To investigate changes in left ventricular (LV) function during exercise in patients with left bundle branch block (LBBB), 22 patients without a history or physical findings of previous myocardial infarction or LV dysfunction were studied by gated radionuclide ventriculography (GRNV) at rest and during bicycle exercise. Coronary arteriography demonstrated greater than 75% diameter narrowing of at least one coronary artery in nine patients. Of the remaining 13 patients, GRNV demonstrated wall motion abnormalities in seven patients either at rest or with exercise. During exercise, mean ejection fraction (EF) did not increase in patients without coronary artery disease (CAD). Patients with CAD had a 12-point fall in mean EF with exercise. We conclude that LV reserve, as demonstrated by ability to increase EF with exercise, is impaired in patients with LBBB even in the absence of CAD or other underlying cardiac disease and that standard GRNV criteria to exclude the presence of CAD (a greater than five-point increase in EF with exercise and normal wall motion) are not strictly applicable in screening patients with LBBB. PMID- 6295129 TI - Isolated guinea pig adrenocortical cells in vitro: morphology and steroidogenesis in control and ACTH-treated cultures. AB - Isolated guinea pig adrenocortical cells were maintained in long-term culture in order to perform sequential experiments on the same cell populations. The cells produced fluorogenic steroids, shown by thin-layer chromatography to be at least aldosterone, cortisol, and corticosterone. In addition, they increased production of these steroids when treated with either ACTH or dibutyryl cyclic AMP. Of particular interest was the fact that cultures treated for the initial 24-hour culture period with ACTH maintained enhanced levels of secretion for several days in absence of hormone and had an enhanced response to ACTH later in the culture period. Such enhancement of secretion was not seen following early treatment with dibutyryl cyclic AMP. The fine structure of the ACTH-treated cells was consistent with increased steroidogenesis. They possessed more smooth-surfaced endoplasmic reticulum, larger mitochondrial crystal surfaces, and larger Golgi complexes than the cells in untreated cultures. PMID- 6295130 TI - National Institutes of Health consensus development conference statement: defined diets and childhood hyperactivity. PMID- 6295131 TI - Bioavailability of iron from wheat bran in pigs. AB - Iron anemia was induced in pigs immediately after birth by feeding an iron depletion diet containing only 17 mg iron/kg feed. (The requirement for iron in this period is 50 mg iron/kg feed). When Hb concentrations were 5 g/100 ml the pigs were given iron repletion diets. One group received 7% bran in the diet, about 60% of the iron derived from the bran and 40% from ferrous sulfate. The other group received no bran and 80% of the iron from ferrous sulfate. There were no differences, either in the increase of Hb or in the increase of serum iron, in the two groups. In a second experiment, one group received all their iron from cereals, and an addition of 20% bran in the diet. The other group received no bran and 80% of the iron from ferrous sulfate. There was no significant difference in the bioavailability of the iron present in the diets. In our experiments bran seemed to have no inhibitory effect on iron absorption, even when 20% bran was included in the diet. PMID- 6295132 TI - Effect of coarse wheat bran fiber and exercise on plasma lipids and lipoproteins in moderately overweight men. AB - The effects of the American Association of Cereal Chemists coarse wheat bran fiber and exercise were evaluated in 20 males, aged 24 to 35 yr, and at least 10% above ideal weight. Thirteen of the subjects participated in a 4 mile walk-jog run program 3 times weekly. In a cross-over design with diets switched midway through the experiment, both the exercising and sedentary groups consumed isocaloric amounts of either white bread or white bread containing wheat bran (0.5 g/kg body weight). Plasma total and lipoprotein cholesterol and triglycerides, body weights, and percentage body fats were determined at 0, 6, and 12 wk. No consistent effects were observed as a result of the wheat bran feeding on any of the parameters measured. Exercise training tended to decrease body weights and percentage body fats, and was associated with significantly increased (p less than 0.05) plasma high-density lipoprotein cholesterol levels and high-density lipoprotein cholesterol to low-density lipoprotein cholesterol ratios. After a rise in triglycerides at wk 6, a significant decline (p less than 0.05) back to base-line values was observed at wk 12 for the exercised subjects. PMID- 6295133 TI - Metachronous bilateral Wilms' tumor, National Wilms' Tumor Study. AB - Eighteen of the patients randomized for treatment according to the first and second National Wilms' Tumor Studies have developed clinically evident tumor in the contralateral kidney subsequent to the initial diagnosis. Only seven of 18 had an adequate exploration of the opposite kidney during the initial surgery and of these seven, four had abnormal findings. Thirteen of 18 cases showed some form of nephroblastomatosis histologically. The original tumors in six and possibly seven were multicentric in origin. While metachronous Wilms' tumors are an unusual occurrence, the prognosis is ominous with only seven of 18 surviving free of disease. The outlook might be improved with adequate attention to the surgical and histologic details during the initial diagnostic procedures. PMID- 6295134 TI - Acute leukemia after successful chemotherapy for oat cell carcinoma. AB - A report of acute myelomonocytic leukemia following successful therapy for oat cell carcinoma is presented. The patient had been treated with extensive cytotoxic and radiation therapy, and was without clinical evidence of disease at one year follow-up. Eighteen months later, a peripheral smear revealed numerous blasts with monocytoid characteristics. This unusual presentation is discussed and compared with several other cases appearing in the recent literature. PMID- 6295135 TI - Thick (myosin) filaments in a glomus tumor. AB - An otherwise classic digital glomus tumor is presented with the unusual ultrastructural finding of cytoplasmic thick (myosin) filaments together with thin (actin) filaments in many of the cells. In places, sarcomere-like orientation was seen. It is little appreciated among diagnostic pathologists, but is well-established, that thick (myosin) filaments occur in smooth muscle type cells. They are present in vivo and can be demonstrated ultrastructurally if rather stringent preparative conditions are met. Whether or not the contractile process in smooth muscle is analogous to skeletal muscle is a debated issue. In the context of diagnostic electron microscopy, it is stressed that thick filaments are not, as often stated, pathognomonic of skeletal muscle neoplasms, and may potentially be found in smooth muscle neoplasms and neoplasms of related cell type (glomus tumors, hemangiopericytomas, tumors of myofibroblasts, etc.). PMID- 6295136 TI - An immunoperoxidase investigation of S-100 protein in granular cell myoblastomas: evidence for Schwann cell derivation. AB - Five cases of granular cell myoblastoma have been studied for detection of the neuroectodermal protein S-100. Immunoperoxidase staining on paraffin sections, using an antibody raised against calf brain S-100 protein, was utilized to demonstrate positive cytoplasmic and nuclear reactivity in all cases. Negative staining in adjacent muscle and connective tissue elements was contrasted to in situ control staining of Schwann cells in peripheral nerves and staining of Langerhans cells and melanocytes in overlying stratified epithelia. These observations are interpreted as support for possible Schwann cell origin of granular cell myoblastomas. PMID- 6295137 TI - C2 deficiency vasculitis: complication of enterocolitis, cutaneous ulcers, and neuropsychiatric disorder. AB - A rare case of systemic vasculitis with second component of complement (C2) deficiency was documented in a patient who developed colonic ulcerations, jejunal edema and dilatation, cutaneous ulcers, peripheral neuropathy, and psychosis. Colonoscopy revealed typical features of ischemic colitis and radiological examination showed ischemic changes in the jejunum and ileum. Histopathological examination of the cutaneous biopsy revealed typical necrotizing vasculitis. It is very likely that multiorgan involvement, including ischemic changes of the intestine, developed secondary to vasculitis associated with C2 deficiency. PMID- 6295138 TI - The role of endoscopy in suspected amebiasis. AB - The diagnosis of amebic colitis can be difficult and confusing. The gross endoscopic appearance as well as the results of endoscopic biopsy can be extremely helpful in differentiating amebiasis from other forms of colitis. Clinical symptoms, laboratory studies, x-ray findings, cultures, and even serological studies may not be sufficient for making an accurate diagnosis. To illustrate the potential difficulties we are reporting three patients in whom the diagnosis of amebiasis was considered but in whom endoscopy was important for arriving at the correct diagnosis. PMID- 6295139 TI - Skeletal muscle uptake of Tc 99m pyrophosphate in amyloidosis. PMID- 6295141 TI - Arsenic--state of the art. AB - Approximately 1.5 million workers in the United States are exposed to arsenic. Occupational exposure is primarily by inhalation. NIOSH recommends that time integrated exposure to arsenic in air not exceed 2 micrograms/m3. Recent exposure is accurately measured by urine assay; urine arsenic concentrations above 50 micrograms/liter indicate increased absorption. Hair assay is a semiquantitative index of past exposure. Toxicity is associated primarily with the trivalent (3+) form of arsenic. Acute poisoning is caused most commonly by contaminated food or drink; it is rarely occupational. Chronic intoxication is characterized by dermatitis, hyperpigmentation, keratoses, peripheral neuropathy (primarily sensory), irritation of the upper and lower respiratory tract, and occasionally by hepatic toxicity and peripheral vasculopathy (blackfoot disease). Arsenic is not carcinogenic in animal species, but is mutagenic in Syrian hamster cells. In man, arsenic is known definitely to cause cancer of skin, lung, and liver (angiosarcoma) and possibly to cause lymphoma. PMID- 6295140 TI - An alternate method for demonstration of bisphosphoglyceromutase (DPGM) on starch gels. AB - The phosphatase activity of bisphosphoglyceromutase (DPGM) was used to determine the phenotypes of the enzyme. DPGM was polymorphic in four Alaskan ethnic groups. PMID- 6295142 TI - Delayed onset sandstone pneumoconiosis: a case report. AB - An unusual case of silicosis is described in a worker who inhaled the dust of pure silica while working in a sandstone quarry. The exposure lasted only eight years. In 1980, 45 years after exposure ceased, severe clinical manifestations of silicosis appeared for the first time. The chest X-ray showed a pneumoconiosis A 2mn/A2 Mn Cor, em, hilus, based on the International Classification of Geneva, 1958. A diagnosis of sandstone pneumoconiosis was made. The case is one further example of late-occurring disease appearing after a latency of several decades. PMID- 6295143 TI - Amphibole fibers in a taconite mine and in the lungs of the miners. AB - Fibers of the amphibole mineral series have been demonstrated in the dust from an open taconite ore mine. Though the total dust levels in some places are high, exposures to fibers are below one fiber/cc. The energy dispersive X-ray spectra of the amphibole fibers correspond to those of cummingtonite-grunerite, hornblende, or actinolite. The same type and size distribution of fibers were found during post mortem analyses of lung tissue from two previously exposed miners. The pathological examination revealed an undifferentiated small cell carcinoma of left lung as well as pulmonary fibrosis in one of the cases. In the second case a poorly differentiated squamous cell carcinoma of left lung was found along with silicosis. PMID- 6295144 TI - Host rocks and gangue minerals in relation to pneumoconiosis and cancer. PMID- 6295145 TI - Aplastic anemia and non-A, non-B hepatitis. AB - Severe aplastic anemia is a rare but important complication of hepatitis. The agent(s) responsible for the hepatitis in these cases have not been well defined. Sixteen patient with hepatitis-associated aplastic anemia were studied for evidence of recent infection with hepatitis A virus, hepatitis B virus, cytomegalovirus, Epstein-Barr virus, and Toxoplasma. Results were compared with data from 10 randomly selected patients with aplastic anemia unassociated with hepatitis. Of the 16 patients, recent acute hepatitis A infection could be excluded in at least 14 patients. Hepatitis B surface antigen (HBsAg) was present in only one patient. A diagnosis of recent hepatitis B infection could not be excluded with confidence in two others. Tests for cytomegalovirus, Epstein-Barr virus, and Toxoplasma gave negative results. No patient with aplasia unassociated with hepatitis had evidence of recent hepatitis A infection, and the frequency of hepatitis B antibodies in this group was indistinguishable from that in patients with hepatitis. These data indicate that most cases of hepatitis that preceded aplastic anemia were not caused by hepatitis A virus or hepatitis B virus; non-A, non-B agents were probably involved in at least 13 of the 16 cases studied. PMID- 6295146 TI - Ketoconazole in the treatment of fungal infection. Clinical and laboratory studies. AB - Ketoconazole is an effective treatment for chronic superficial candidiasis as well as chronic dermatophytosis. In the latter group of infections the best results were obtained in patients with tinea corporis who were not responsive to griseofulvin. It is possible to maintain some patients with chronic mucocutaneous candidiasis in remission without using prophylactic ketoconazole, although relapses may occur. However, the responses of patients with Hendersonula and Scytalidium infections as well as those with subcutaneous mycoses, such as eumycetoma, were disappointing. Patients who have an inadequate response to ketoconazole may also have subnormal serum levels of the drug and the value of such estimations in routine management needs further evaluation. PMID- 6295147 TI - The mechanism of action of the new antimycotic ketoconazole. AB - Ketoconazole is one of the new members of the imidazole series with a broad spectrum antifungal profile. Although sharing its basic active principles with the other imidazoles, ketoconazole obtains its superior in vivo activity mainly from its good oral absorption and its lower degree of inactivation once absorbed. Its selective toxicity for yeasts and fungi is found to be primarily linked to the inhibition of ergosterol biosynthesis and to interference with other membrane lipids. In vitro growth studies revealed that ketoconazole's activity was more pronounced against the invasive morphogenetic form than against the saprophytic form of Candida albicans, which at least partly explains its prominent in vivo potency. At extremely low concentrations (10 ng/ml-1) ketoconazole prevents the development of the very form that is responsible for the expression of clinical symptoms. In contrast to other imidazoles, ketoconazole's action on the morphogenesis of the organism is not influenced by serum. The synergistic action with host defense cells, as demonstrated in culture systems, is another inherent property of this drug and may have a great impact on the eradication of systemic fungal infections. These effects of ketoconazole have been studied in a variety of fungal organisms with the aid of phase-contrast, scanning, and transmission electron microscopy in order to characterize ketoconazole's profile in comparison to the other imidazole derivatives. PMID- 6295148 TI - Therapy of chronic mucocutaneous candidiasis. AB - Clinical observations made on a group of six children and three adults indicate that ketoconazole is a safe and effective fungistatic agent that is useful for both initial and long-term maintenance therapy of mucocutaneous lesions due to infection by sensitive strains of Candida in patients with chronic mucocutaneous candidiasis. PMID- 6295149 TI - Long-term therapy of chronic mucocutaneous candidiasis with ketoconazole: experience with twenty-one patients. AB - Our experience in the treatment of chronic mucocutaneous candidiasis with ketoconazole is reviewed. Of 21 patients, 15 have evidence of deficient cellular immunity and eight have endocrine abnormalities. Six patients had concurrent dermatophytosis or chromomycosis. All patients responded to treatment. Mucosal lesions improved in 6.7 +/- 0.5 days and cutaneous lesions responded to 22.7 +/- 5.1 days. The responses by infected nails were more variable (mean response time 92.4 +/- 14.4 days). Concurrent dermatophytoses did not prolong response times. Adverse effects were infrequent: one patient had drug-induced hepatitis and two patients became hypertensive. The relationship of hypertension to ketoconazole treatment is unclear. One patient was able to remain in remission after treatment was discontinued. Two patients had relapses while on treatment. Candida albicans isolated from these patients was highly resistant to ketoconazole in vitro. We conclude that ketoconazole is an effective and well-tolerated drug for the treatment of the infectious component of chronic mucocutaneous candidiasis. PMID- 6295150 TI - Laboratory and clinical assessment of ketoconazole in deep-seated mycoses. AB - Forty-eight cases of deep mycoses were studied and treated with ketoconazole, each with in vitro evaluation of the minimum inhibitory concentrations (MIC) of the causative fungi, in vivo pharmacokinetic, clinical, and mycologic evaluations, several months to two years after the treatment was stopped. Excellent results were obtained in six cases of chronic mucocutaneous candidiasis, with restoration of immunologic disturbances; 23 cases of systemic candidiasis, including new aspects of heroin addicts with cutaneous, ocular, or osteoarticular manifestations; eight cases of histoplasmosis, five due to Histoplasma capsulatum and three to Histoplasma duboisii, with cure in seven and remission in one; one case of African blastomycosis (Blastomyces dermatitidis); three cases of mycetoma, two due to Monosporium apiospermum, one due to a dematiacious fungus; three cases of entomophthoromycosis with cure; one case of fungal arthritis, due to new hyphomycete similar to M. apiospermum, pathogenic for laboratory animals; one case of Drechslera longirostrata causing vertebral arthritis, following a fungal endocarditis and cured by combination of ketoconazole with amphotericin B, each agent alone being ineffective; and other deep mycoses. PMID- 6295151 TI - Treatment of paracoccidioidomycosis with ketoconazole: a three-year experience. AB - The results of ketoconazole therapy in 38 patients with active paracoccidioidomycosis are described. Treatment consisted of a 200 mg tablet a day for 6 months. Evaluation was accomplished by means of a scoring system and the results were as follows: none of the patients worsened during therapy, one was found to be unchanged, five had minor improvement, 330 had major improvement, and there was complete resolution of the pretherapy conditions in 13. These findings plus the lack of toxicity of the drug and the facility for oral administration, make of ketoconazole a first line drug for the treatment of paracoccidioidomycosis. PMID- 6295152 TI - Post-therapy status of paracoccidioidomycosis treated with ketoconazole. AB - Twenty-four paracoccidioidomycosis patients who completed a course of therapy with ketoconazole were followed for 12 months after treatment, and 10 of these patients were followed for 24 months. Only two of these patients relapsed; the remaining continued in remission. There were no fatalities. Immunologic tests showed decreased antibody activity and increased skin test reactivity to paracoccidioidin in comparison with the results obtained at the end of therapy. These findings indicate that ketoconazole therapy has improved the prognosis of patients with paracoccidioidomycosis. PMID- 6295153 TI - Experience with ketoconazole in three major manifestations of progressive coccidioidomycosis. AB - Forty-seven courses of oral ketoconazole therapy administered to 46 patients with progressive coccidioidomycosis were evaluated. Responses, of varying degrees evaluated by a point system, occurred in 81, 94, and 91 percent of courses evaluated for skeletal, chronic pulmonary, and cutaneous disease, respectively. Presumed side effects occurred in 26 percent of courses and were generally minor and reversible. Relapse occurred in 33, 33, and 11 percent of evaluable responding patients with skeletal, chronic pulmonary and cutaneous disease, respectively. From this experience, ketoconazole appears to be an important advance in the therapy of chronic coccidioidomycosis. Whether cure is possible will require further study and follow-up. PMID- 6295155 TI - Treatment of disseminated and progressive cavitary histoplasmosis with ketoconazole. AB - Ten patients with disseminated histoplasmosis and seven with progressive cavitary histoplasmosis were treated with ketoconazole, an imidazole derivative that is well absorbed orally and relatively nontoxic. Seven of seven noncompromised hosts with disseminated disease tolerated therapy well and achieved clinical and mycologic cures. Although well tolerated in all three compromised hosts with disseminated disease, none achieved clinical and bacteriologic cures. All patients with progressive cavitary disease tolerated therapy well and six of seven achieved clinical and radiographic cures. Therefore ketoconazole appears safe and effective in the treatment of disseminated or progressive cavitary histoplasmosis in the noncompromised host. In the compromised host with disseminated histoplasmosis, ketoconazole does not appear to be effective and more conventional therapy appears warranted. PMID- 6295154 TI - Treatment of coccidioidomycosis with ketoconazole: an evaluation utilizing a new scoring system. AB - The evaluation of the response of patients with coccidioidomycosis to any therapeutic modality is a major challenge. A numerical scoring system was devised to quantitate separately the severity of disease on clinical presentation, the findings on chest film, bone scan, gallium scan, serology and skin test with coccidioidin and spherulin. The scoring system was used to evaluate the response to treatment with ketoconazole of seven patients with infiltrate pulmonary coccidioidomycosis; 20 patients with chronic cavitary coccidioidomycosis; and 40 patients with disseminated coccidioidomycosis. Dissemination included the soft tissue in 15, bone in 15, synovium in 11 and skin in 18. In all categories clinical severity scores improved dramatically. Radiographic scores showed similar improvement in cases of infiltrative pulmonary coccidioidomycosis but showed no change in cavitary coccidioidomycosis. Serology scores improved significantly (-2 or more) in one of seven infiltrative pulmonary cases, three of twenty chronic cavitary cases and twenty-three of forty disseminated cases. Among those with adequate mycology followup, cultures converted to negative in two of three infiltrative pulmonary coccidioidomycosis; seven of fourteen chronic cavitary coccidioidomycosis; and sixteen of twenty-two with disseminated disease. Unfortunately, when ketoconazole was discontinued or interrupted, symptoms recurred in four of twenty (20 percent) with chronic cavitary and ten of forty (25 percent) of disseminated cases. The disease in two patients progressed while on ketonconazole. One of those developed meningitis. PMID- 6295156 TI - Treatment of mycoses in cancer patients. AB - Invasive fungal infections are becoming increasingly frequent among immunocompromised patients and especially among cancer patients. The most common pathogens identified are Candida species, Aspergillus species, Cryptococcus neoformans, and Mucor species. Amphotericin B remains the mainstay of antifungal therapy. However, the toxicity of this drug may limit its use and, in addition, both failures and relapses have been reported. 5-Fluorocytosine and imidazoles, such as miconazole and ketoconazole, have been shown to be active, mainly on yeast organisms. The emergence of 5-fluorocytosine-resistant strains warrants caution for its administration as a single agent. The specific role of ketoconazole has not yet been established in large studies. In our experience, ketoconazole seems to be effective in the treatment of severe oral candidiasis in non-neutropenic cancer patients. Moreover, ketoconazole administered prophylactically to neutropenic patients decreases the number of positive surveillance cultures in these patients. The rare incidence of major toxicity and the ability to administer ketoconazole orally represent also major arguments for further investigation of ketoconazole activity by prospective controlled studies. PMID- 6295157 TI - Potential and problems with ketoconazole. PMID- 6295159 TI - The Greig cephalopolysyndactyly syndrome in a Canadian family. PMID- 6295158 TI - The antifungal activity of ketoconazole. AB - The antifungal properties of ketoconazole were investigated both in vitro and in vivo. The antifungal potency of ketoconazole in vitro was studied in Sabouraud's broth for 715 fungal strains belonging to 85 species and several strains were tested in other media, including Eagle's minimal essential medium. Ketoconazole is highly active in vitro and possesses broadspectrum activity. Its in vitro activity is largely dependent on the medium used. Ketoconazole's activity is increased in medium enriched with serum and in Eagle's minimal essential medium. Ketoconazole is very potent in the topical treatment of skin dermatophytosis, skin candidiasis, and in vaginal candidiasis of laboratory animals. Ketoconazole is superior to griseofulvin in the oral treatment of skin dermatophytosis. Furthermore, ketoconazole is orally highly active in skin candidiasis in guinea pigs, in vaginal candidiasis in rats, and in gastrointestinal candidiasis in various animal animal species. In systemic candidiasis and in disseminated dermatophytosis in guinea pigs cure with oral ketoconazole is achieved. No side effects are observed. PMID- 6295161 TI - Participation of the renin system in treatment-resistant hypertension as measured by acute blockade of the angiotensin converting enzyme. AB - The acute blood pressure response to the angiotensin converting enzyme inhibitor, captopril, was used to measure participation of the renin-angiotensin system in treatment-resistant hypertension. By 2 h after a single 25-mg oral captopril dose in patients still receiving a diuretic-vasodilator-beta-blocker combination, systolic and diastolic blood pressures had fallen significantly; the decrease in diastolic pressure correlated with the control (immediately pre-captopril) plasma renin activity (r = 0.64, p less than 0.001). Apart from its possible contribution to the underlying hypertension, this captopril-identified renin component may have reflected diuretic and vasodilator-induced renin stimulation that could not adequately be prevented by the renin-lowering properties of the beta-blocker. Captopril, and perhaps other specific antirenin drugs, therefore, may be valuable adjuncts to treatment in patients with hypertension refractory to conventional therapy. PMID- 6295160 TI - Urinary calcium excretion at extremes of sodium intake in normal man. AB - To elucidate the relationship between the renal regulation of sodium and calcium excretion at extremes of sodium intake, we studied 6 normal men ingesting a fixed, 400 mg/day calcium intake and four levels of sodium intake from 10 to 1,500 mEq/day. Serum ionized calcium was not influenced by sodium intake. Blood pressure and cardiac index increased modestly. Urinary calcium excretion increased to 262 +/- 53 mg/day (mean +/- SE). Plasma norepinephrine concentration, serum PTH levels, hematocrit, total serum protein concentrations and CO2 content decreased with increasing sodium intake. Urinary cAMP increased as sodium intake was raised from 10 to 300 mEq/day, but subsequently decreased to basal values. Urinary calcium and sodium excretion were related (p less than 0.001) in a nonlinear fashion as were the fractional excretions of these cations (p less than 0.001). The filtered calcium load and the fractional calcium excretion were directly and linearly related (p less than 0.001). We conclude that the effect of sodium intake on urinary calcium excretion principally reflects changes in the filtered calcium load rather than changes in renal sodium handling. Calcium homeostasis at extremes of sodium intake does not appear to be critically dependent upon PTH-mediated mechanisms. The data suggest that the proximal tubule has a remarkable capacity to dissociate calcium resorption from that of sodium. PMID- 6295162 TI - Increase in plasma beta-endorphin-like immunoreactivity at parturition in normal women. PMID- 6295164 TI - Experimental primary cytomegalovirus infection in pregnancy: timing and fetal outcome. AB - In contrast to intrauterine rubella infection, the relationship between timing of maternal cytomegalovirus (CMV) infection and fetal outcome has not been clearly defined. In order to investigate this relationship, a guinea pig model was utilized to assess the fetal consequences of maternal CMV infection during the first, second, or third trimester of pregnancy. Congenital infection occurred in 24 of 35 newborn guinea pigs (69%) delivered to mothers infected during the third trimester, with localization of virus to salivary gland in 17 of the 24 infected newborn guinea pigs. In contrast, only one of 28 (5%) progeny sacrificed following first-trimester maternal infection was congenitally infected (p less than 0.01). Second-trimester maternal infection was associated with an intermediate risk of intrauterine infection with transmission of virus to 17 of 54 progeny (33%) (p less than 0.01). Eight of the 10 fetuses delivered after second-trimester infection had virus in multiple organs including the brain. These data suggest that timing of maternal CMV infection is an important variable affecting fetal outcome, with increased risk of intrauterine infection when maternal infection occurs late in pregnancy. However, if fetal infection occurs earlier in pregnancy, it appears to present a greater threat to the fetus, with the potential for dissemination of virus in multiple fetal tissues, including the brain. PMID- 6295163 TI - Observation of an organism found in patients with gestational trophoblastic disease and in patients with toxemia of pregnancy. AB - This is an initial descriptive report of observations of multiple forms of an organism found in patients with gestational trophoblastic disease and in patients with preeclampsia-eclampsia. The worm-like forms most frequently observed have an average length of 1.0 to 1.5 mm. Larva-like forms have an average length of 150 mu; primordial eggs and egg-like forms in developmental stages range from 7 to 43 mu in diameter; and sperm-like forms are 3.5 mu or slightly smaller in size. These forms have been observed in contact smears prepared from 3 ml samples of peripheral circulating blood from both groups of patients, from trophoblastic tumor tissue, from contact smears prepared from placentas of patients with preeclampsia-eclampsia, and from umbilical cord blood of infants delivered of patients with preeclampsia-eclampsia. The various forms of this organism share morphologic characteristics of several orders of helminths, i.e., hookworms, roundworms and tapeworms. The taxonomy of these forms has not yet been determined. Until the time of taxonomic classification, the various forms will be referred to as Hydatoxi lualba. We have experimental evidence that this organism has biologic activity in BALB/c mice and in beagle dogs. PMID- 6295165 TI - A herpesvirus antigen in human premalignant and malignant cervical biopsies and explants. AB - Cervical biopsies and explant cultures from patients with squamous metaplasia, various grades of dysplasia, carcinoma in situ (CIS), and invasive squamous cell carcinoma were screened for VP143, an early nonstructural polypeptide of herpes simplex virus type 2 (HSV-2), VP143 was identified in 31% of biopsies exhibiting severe dysplasia, 29% with CIS, and 41% with invasive squamous cell carcinoma. Similar results were obtained when explants derived from these biopsies were examined for VP143. The expression of the protein persisted in passaged subcultures in four of five invasive carcinomas which originally contained VP143. Staining for VP157, the major capsid protein of HSV-2, was absent. Furthermore, virus structures were not seen by electron microscopy and infectious virus was not isolated from cell cultures inoculated with biopsy extracts. These results suggest that VP143 was expressed in the premalignant and malignant cervical cells in the absence of productive viral infection. Thus, a fragment of the HSV-2 genome was retained within the cells, the expression of which resulted in the production of VP143. PMID- 6295166 TI - Ovarian function after radical hysterectomy for stage IB carcinoma of cervix. AB - Ovarian function was assessed in 20 patients after radical hysterectomy and lymph node dissection for Stage IB cervical carcinoma. All patients were under 45 years of age, and four were or had been on estrogen therapy for postmenopausal symptoms. The other 16 patients were free of symptoms and demonstrated premenopausal gonadotropin profiles. Fourteen of these 16 had luteal phase serum progesterone levels. Only one patient required reoperation for a pathologic condition of the adnexa. A surgical approach to Stage IB cervical carcinoma conserves ovarian function in 80% of patients. PMID- 6295167 TI - Follicle-stimulating hormone--regulated granulosa cell steroidogenesis: involvement of the calcium-calmodulin system. AB - The role of the calcium-calmodulin system in the gonadotropic regulation of ovarian steroidogenesis was studied by investigating the influence of various agents known to alter calcium metabolism or calmodulin activity on basal and follicle-stimulating hormone (FSH)-stimulated production of progesterone by rat granulosa cells in vitro. Lanthanum, a specific calcium antagonist, attenuated FSH-stimulated cyclic adenosine monophosphate (cyclic AMP) and progesterone production. [Ethylene-bis(oxyethylene-nitrilo)]tetraacetic acid (EGTA) significantly reduced this steroidogenic response but failed to alter the synthesis of the nucleotide. Although progesterone production was markedly increased by dibutyryl cyclic AMP, this was significantly lowered by an inhibitor of calcium uptake, verapamil. FSH-stimulated production of cyclic AMP and progesterone and dibutyryl cyclic AMP-induced progesterone biosynthesis were all significantly reduced by trifluoperazine, a specific inhibitor of calmodulin. These results indicate the existence of at least two sites in the trophic regulation of granulosa cell steroidogenesis which are calcium- and calmodulin dependent: one localized at the level of cyclic AMP production, and the second at an unidentified step(s) distal to the formation of this nucleotide. PMID- 6295169 TI - Combined clinical and computed tomographic diagnosis of primary lacrimal fossa lesions. AB - We studied 39 patients who had solid mass-lesions primary in the lacrimal gland by computed tomography and reviewed their clinical histories. Twenty-three patients had either inflammatory conditions (16 cases) or lymphoid tumors (seven cases), with average symptomatic periods of less than a year. In this group, soft tissue contour analysis in the axial and corneal projections demonstrated diffuse, compressed, and molded enlargements of the lacrimal gland in an oblong fashion, and there were no associated bone defects. Sixteen parenchymal benign or malignant tumors (six benign mixed tumors, one schwannoma, and nine malignant epithelial tumors) exhibited rounded or globular soft-tissue outlines and were frequently associated with contiguous bone changes. The benign tumors had smooth encapsulated outlines at their margins, whereas the malignant tumors displayed microserrations indicative of infiltration. The patients with the benign mixed tumors had had symptoms, on the average, for more than a year, whereas those with epithelial malignancies became symptomatic or had a preexisting benign mixed tumor that became exacerbated in periods of less than six months. Contour analysis of the soft-tissue mass depicted in coronal and axial tomograms is a valuable adjunct that leads to more accurate preoperative diagnosis when combined with a radiographic search for bone changes and the clinical history. Once a diagnosis regarding the presumptive lesional family has been made preoperatively, corticosteroid therapy may be instituted for acute inflammation and biopsies through the eyelid should be performed for suspected chronic inflammations, lymphoid lesions, or epithelial malignancies. A lateral orbitotomy without prior biopsy should be performed for rounded, well-encapsulated masses of long duration that are likely to be benign mixed tumors. PMID- 6295170 TI - An analysis of retinal cotton-wool spots and cytomegalovirus retinitis in the acquired immunodeficiency syndrome. PMID- 6295168 TI - Steroid modulation of pregnenolone to progesterone conversion by human placental cells in vitro. AB - Studies were performed to examine the production of progesterone by human placental cells in vitro. Samples of placentas from 22 women at term after spontaneous onset of labor and vaginal delivery were utilized. The tissue was dispersed into isolated cells with the use of collagenase, and suspensions of these cells were incubated with pregnenolone as substrate in the presence or absence of other compounds which may regulate progesterone production. These cell preparations produced progesterone in a dose-related fashion with exogenous pregnenolone. The conversion of pregnenolone to progesterone occurred rapidly, with most of the conversion completed during the first hour of incubation. The conversion was inhibited by dehydroepiandrosterone, estrone, androstenedione, and testosterone (p less than 0.001 in all cases). The inhibitory effect of androstenedione and testosterone was not dependent on aromatization to estrogen. Dihydrotestosterone and 5 alpha-pregnanedione resulted in a significant increase in the amount of progesterone present (p less than 0.001). In preliminary experiments, gonadotropin-releasing hormone, salbutamol, and propranolol were without significant effect in this system. We conclude that this system is a useful model for studying progesterone production by human placental tissue, and that placental progesterone production may be significantly influenced by the presence of other steroid hormones. PMID- 6295171 TI - Ocular histopathologic studies of neonatal and childhood adrenoleukodystrophy. AB - Histopathologic studies of the eyes of one patient (a boy who died at 14 years of age) with childhood adrenoleukodystrophy and two patients (girls who died at 24 and 31 months of age) with neonatal adrenoleukodystrophy showed the accumulation of the characteristic bileaflet inclusions in optic nerve macrophages, retinal neurons, and macrophages and loss of ganglion cell and nerve fiber layer. Additionally, in the two cases of neonatal adrenoleukodystrophy, changes resembling early retinitis pigmentosa were found, with accumulation of characteristic inclusions in the retinal pigment epithelium and pigment-laden macrophages. One of the patients with neonatal adrenoleukodystrophy also had an anterior subcapsular cataract and cystoid macular edema. PMID- 6295172 TI - Studies on the pathogenesis of avian rickets II. Necrosis of perforating epiphyseal vessels during recovery from rickets in chicks caused by vitamin D3 deficiency. AB - This study involved comparison of the distribution and integrity of perforating epiphyseal and marrow vessels with the stage of development and integrity of chondrocytes and the distribution of insoluble calcium in the proximal tibial growth plate of 3-week-old vitamin-D3-deficient hypocalcemic chicks and 3-week old D3-deficient chicks 12, 36, 72, and 120 hours after an oral dose of 10,000 units vitamin D3. The aim was to clarify the mechanisms responsible for chondrocyte hypertrophy and cartilage calcification in the avian growth plate. Within 12 hours after administration of vitamin D3, serum calcium levels rose to normocalcemic levels. The following morphologic changes were first recognizable at the times indicated. Distal portions of previously elongated perforating epiphyseal vessels and adjacent proliferative and maturing chondrocytes underwent necrosis by 12 hours. Chondrocyte necrosis was not preceded by hypertrophy. By 36 hours, vascular and chondrocyte necrosis involved large portions of the thickened proliferating and maturing zone, and perforating epiphyseal vessels were shortened to a normal length. By 72 hours, chondrocyte hypertrophy and calcification resumed around the shortened epiphyseal vessels. By 120 hours, marrow had removed the necrotic cartilage, and morphologically normal growth plate was restored, with perforating epiphyseal and marrow vessels, both ending in a narrow hypertropic cartilage zone. The results indicate that proximity of chondrocytes to perforating epiphyseal vessels is necessary for their viability, but loss of these vessels does not cause hypertrophy. Since hypertrophy and calcification both occur in the proximity of perforating epiphyseal vessels in normocalcemic animals but not in hypocalcemic animals, it is likely that the vessels influence hypertrophy and calcification by delivering calcium to chondrocytes. PMID- 6295173 TI - Macroglobulinemic neuropathy. Experimental macroglobulinemic neuropathy in mice. PMID- 6295174 TI - Experimental silicosis. II. Long-term effects of intratracheally instilled quartz on collagen metabolism and morphologic characteristics of rat lungs. AB - Rats received intratracheal instillations of 50 mg of silica (quartz, 0.5 mu particles). One, 2, 4, 5, 6, 9, and 12 months later, the lungs were evaluated histologically and by various biochemical measurements. The lung content of protein, proline, and hydroxyproline (collagen) were quantitated, as were the synthesis rates of lung collagen and the total lung protein (evaluated with lung minces in vitro. The ratio of newly synthesized and of total lung Type I to Type III collagen was also determined. These experiments were performed in parallel on rats free of chronic respiratory disease and a strain of conventional animals. The authors conclude that 1) the excess collagen deposited in granulomas and/or silicotic nodules as part of the fibrotic response of the lung is similar to normal lung collagen with respect to relative ratios of Types I and III present, in contrast to the response of the lung to oxidant pneumotoxins; 2) the response of the lung to silica continues for at least 1 year; 3) there are essentially no differences in the response of chronic respiratory disease-free Sprague-Dawley and conventional Wistar rats to intratracheally instilled silica. Both strains of rats develop silica-containing granulomas, mature silicotic nodules, and areas of alveolar lipoproteinosis associated with interstitial pneumonitis. Even 1 year after instillation of silica areas of granulomas, silicotic nodules and alveolar lipoproteinosis may be observed in most of the lungs studied; ie, these responses are not mutually exclusive. PMID- 6295176 TI - Modulation of intracellular Na+ activity and cardiac force by norepinephrine and Ca2+. AB - The actions of norepinephrine and high calcium on the electrical, mechanical, and intracellular sodium ion activities were studied in electrically driven canine cardiac Purkinje fibers under different conditions. It was found that norepinephrine and high calcium decrease intracellular sodium ion activity (aiNa). The exposure to either agent is followed by a transient decline of force that correlates with the lower aiNa. Inhibition of the Na+ -K+ pump by strophanthidin reduces or abolishes the decrease in aiNa by norepinephrine but not that by high calcium. It is concluded that norepinephrine and high calcium both decrease aiNa and thereby the contractile force but (unlike high calcium) norepinephrine acts through the stimulation of the Na+ -K+ pump. PMID- 6295175 TI - Modification of pancreatic carcinogenesis in the hamster model. 2. The effect of partial pancreatectomy. AB - The effect of partial pancreatectomy (PP) on the pancreatic carcinogenicity of N nitrosobis (2-oxopropyl)amine (BOP) was investigated in Syrian golden hamsters by subcutaneous injection of a single dose of BOP (20 mg/kg, body weight) given 30 minutes after (Group 1), 1 week after (Group 2), or 1 week before 70% PP (Group 3). Additional groups consisted of animals with PP alone (Group 4), sham operation (laparotomy) followed 30 minutes later by BOP treatment (Group 5), and BOP treatment only (Group 6). The experiment was terminated 46 weeks after BOP administration in each group. The pancreas and extrahepatic bile ducts, including the common duct and gallbladder, were examined histologically. Tumor patterns were compared in hamsters with PP and in the corresponding segments of the pancreas in BOP-treated control groups. The pancreatic cancer incidence was highest (31%) in Group 2 and lowest in Group 1 (3%), a difference that was statistically significant (P less than 0.01). Also, a statistically highly significant larger number of tumors occurred in Group 2, compared with group 1, 3, or 5 (P less than 0.0005). In a comparison of the number of carcinomas per tumor-bearing hamster, there were greater numbers of carcinomas in Group 2 (2.6 carcinomas) than in Groups 1, 3, 5, and 6 (1.0, 1.0, 1.3, and 2.6 tumors, respectively). Moreover, pancreatic tumors in Group 2 hamsters were larger (average diameter, 10 mm) than in Group 1 (4 mm), Group 3 (3.5 mm), Group 5 (4 mm), and Group 6 (average, 9mm). The incidence of extrapancreatic tumors did not vary among the PP groups but was equally lower than those in BOP-treated control groups. The data indicated BOP carcinogenesis was inhibited by surgery (regardless of whether PP was per formed) when the carcinogen was given 30 minutes after the surgery but was significantly enhanced when BOP was administered 1 week after PP. The possible reasons for these conflicting results are discussed. Morphologically all tumors were of ductular, ductal, and mixed ductular-insular patterns and most developed at the resected margins, where proliferation of islets, ducts, and ductules, but not of acinar cells, occurred. The results confirm our view that the ductal and ductular cells are the progenitor cells for BOP-induced pancreatic tumors in hamsters. PMID- 6295177 TI - Nonenzymatic separation of myocardial cell nuclei from whole heart tissue. AB - A simple and rapid nonenzymatic method has been developed to isolate myocardial cell nuclei from whole heart tissue. This method consists of a controlled disruption of cells followed by isopycnic gradient centrifugation. We have reviewed and compared our method to others more lengthy and laborious. By using a number of criteria, such as morphometric measurements, chemical composition, functional studies, specific nuclear protein markers, and mathematical analysis, we show that the nonenzymatic digestion method provides a most useful technique for the study of the biochemistry of the myocardial cell nucleus. PMID- 6295178 TI - Effect of glutaraldehyde on hydrosmotic response of toad bladder to vasopressin. AB - The present study investigates the time-, dose-, and temperature-dependence of glutaraldehyde action on the permeability to water of the toad bladder. Bladders preincubated with increasing concentrations of glutaraldehyde become progressively desensitized to the hydrosmotic action of vasopressin (ADH), theophylline, and dibutyryl adenosine 3',5'-cyclic monophosphate (dibutyryl cAMP). The ADH response was reduced by 50% with 0.03% glutaraldehyde applied to the serosal side for 10 min at 4 degrees C. Sixfold higher doses of glutaraldehyde were required with mucosal application. Bladders partially fixed with low-dose glutaraldehyde exhibit a markedly prolonged duration of action of ADH. Bladders fixed with higher doses of glutaraldehyde in the presence of ADH retain a high permeability to water for prolonged periods even in the absence of ADH. This action of glutaraldehyde to stabilize the hormone-induced water channels is also considerably more effective with serosal than with mucosal application. As the rate-limiting permeability barrier for water affected by ADH is known to be located in the apical membrane, these findings suggest that glutaraldehyde exerts its action from an intracellular position. It is postulated that glutaraldehyde stabilizes the ADH-induced channels by cross-linkage of amino groups and other reactive sites at the cytoplasmic surface of the apical membrane and/or by inactivating the intracellular machinery responsible for the dispersal or removal of water channels in the hormone target cell. PMID- 6295179 TI - Voltage-dependent Ca2+ channel and Na+ channel in frog taste cells. AB - Frog taste cells were hyperpolarized by injecting an inward current pulse, and regenerative anode-break potentials were observed at the termination of the current pulse. The results obtained are as follows. 1) The magnitude of the anode break potentials increased with the extent of hyperpolarization of taste cells and reached a saturation level around -200 mV. 2) The magnitudes of the anode break potentials observed in 80 different taste cells hyperpolarized to about 200 mV were distributed widely from cell to cell. The average magnitude was 39 mV. 3) The anode-break potentials were recorded after the lingual artery was perfused with artificial solutions containing various channel blockers. The results indicated that the anode-break potentials are composed of Na+ and Ca2+ components. 4) The slope of the current-voltage relation obtained with cells hyperpolarized to 100 mV was appreciably decreased above -50 mV by application of tetrodotoxin to the perfusing solution. Discussion was made on possible roles of the voltage-dependent Na+ and Ca2+ channels in the electrotonic spreading of the depolarization at the receptor membranes to the synaptic area and in releasing a chemical transmitter. PMID- 6295180 TI - Localization and properties of bovine photoreceptor 5'-nucleotidase. AB - The main light-activated enzyme of the vertebrate photoreceptor is cGMP phosphodiesterase, whose product is GMP. GMP would be broken down to guanosine by the enzyme 5'-nucleotidase on the cytoplasmic (extradiscal) surface of the disks. The presence of 5'-nucleotidase on the cytoplasmic surface was verified by using sucrose continuous gradients to show its association with the photoreceptors and by using disk preparation and concanavalin A binding to demonstrate its presence on the extradiscal surface. Further studies using detergents and freeze-thaw showed that an even higher 5'-nucleotidase activity is present on the intradiscal surface; however, it is the smaller cytoplasmic surface activity that is potentially relevant to the physiology. The 5'-nucleotidase on the extradiscal surface is light insensitive, has a broad optimal pH range, shows a divalent cation dependence, and is competitively inhibited by nucleoside di- and triphosphates. When the data determined experimentally were extrapolated to physiological conditions, we obtained a decay time constant for GMP breakdown by 5'-nucleotidase in the range of 0.4 to 1.06 s. This time constant is in the range of the time constants of the fall of rod cell receptor potential, suggesting a possible role for GMP level in visual transduction. PMID- 6295182 TI - Vasopressin and angiotensin II receptors in rat aortic smooth muscle cells in culture. AB - Rat aortic smooth muscle cells were isolated and maintained in primary culture. After 2-3 days, cells recovered their contractile phenotype and could be induced to contract in response to vasopressin and angiotensin II. Vasopressin- and angiotensin-specific binding sites were detected on these cells, using tritiated Lys8-vasopressin, Asn1-Val5-angiotensin II, and Sarc1-Ile8-angiotensin II. Vasopressin binding sites had Kd values of 30 and 12 nM for Lys8-and Arg8 vasopressin, respectively, and a maximal binding capacity of 25,000 sites/cell. They displayed several of the expected characteristics of vasopressin receptors involved in the vasopressor response in vivo. A highly significant correlation was found between the relative agonistic or antagonistic vasopressor potencies of a series of vasopressin structural analogues and their relative abilities to inhibit [3H]vasopressin binding to aortic smooth muscle cells. Specific binding sites for Asn1-Val5-angiotensin II and Sarc1-Ile8-angiotensin II had the following characteristics: Kd = 2.3 and 1.3 nM, respectively; maximal capacity: 50,000 sites/cell. Vasopressin and angiotensin did not modify the intracellular cyclic AMP content of aortic smooth muscle cells. PMID- 6295181 TI - Stimulus-secretion coupling in beta-cells: modulation by pH. AB - We have examined the influence of changes in pH on the oscillatory pattern of electrical activity (EA) in the beta-cell by altering medium pH (pHo) and using permeable weak buffers to alter intracellular pH (pHi). A decrease in pH in the presence of glucose elicited depolarization to the active phase and constant spike activity, whereas an increase in pH elicited a decrease in spike activity or silent hyperpolarization. On inhibition of HCO3:Cl antiport by addition of DIDS (4,4'-diisothiocyano-2,2'-stilbene disulfonic acid), probenecid, or withdrawal of medium HCO-3, there was an increase in the duration of the active phase. A similar result was obtained on the inhibition of Na:H antiport by the addition of amiloride or the reduction of medium [Na+]. The influence of H+ and glucose has been proposed to decrease K+ permeability (PK). However, the influence of pH on 86Rb+ efflux was most effective at subthreshold or 4.2 mM glucose; only a moderate decrease in PK occurred at 8.3 mM glucose, and no effect was obtained at 16.7 mM glucose. Alteration of pHi, and not pHo, induces similar effects on glucose-induced electrical and secretory events. There is a clear dissociation between the influence of inhibitors of the Na:H and HCO3:Cl antiporters on the electrical and secretory events. DIDS and amiloride increased glucose-induced EA, but markedly inhibited the secretory response to glucose. It is evident that pH modulates the electrical events and cationic fluxes and ultimately influences the transduction of information to the mechanisms controlling the secretory process in the beta-cell. PMID- 6295183 TI - Decreased distal acidification in acute hypercapnia in the dog. AB - The present studies evaluate the effect of acute hypercapnia on distal nephron H+ secretion (DNH+S) in vivo by means of the urine-blood PCO2 difference (U-B PCO2) in alkaline urine. Bicarbonaturia was induced by either a sodium bicarbonate infusion or L-lysine administration. Our results demonstrate that the U-B PCO2, as a function of the urinary bicarbonate concentration, was significantly lower during acute respiratory acidosis; this effect was not dependent on changes in glomerular filtration rate and/or fractional excretion of sodium, potassium, and chloride. Infusion of the sodium salts of sulfate, a nonreabsorbable anion, did not correct the diminished U-B PCO2. Amiloride caused the U-B PCO2 to fall in normocapnic dogs but not in hypercapnic dogs. When hypercapnia was superimposed in dogs with extracellular fluid volume contraction, there were no changes in the U-B PCO2. This study indicates that acute hypercapnia in the intact dog decreases DNH+S and is compatible with an effect of hypercapnia on the voltage-dependent component of urine acidification. The mechanism appears to be direct rather than secondary to factors that influence the rate of sodium delivery to the distal nephron. PMID- 6295184 TI - Intracellular microelectrode characterization of the rabbit cortical collecting duct. AB - Cortical collecting ducts of the rabbit were perfused in vitro and the intracellular potential (Vbl) was measured with KCl-filled microelectrodes. The ratio of apical to basolateral membrane resistance (Ra/Rbl) was estimated from the voltage divider ratio using cable analysis. In control tubules Vbl averaged- 84.0 +/- 2.5 mV and Ra/Rbl was 0.83 +/- 0.11. Pretreatment of the rabbits with mineralocorticoid caused Vbl to hyperpolarize to--105.8 +/- 3.1 mV and Ra/Rbl to decrease slightly to 0.62 +/- 0.10. A 10-fold increase of the luminal [K+] caused a 40.6 +/- 3.1 mV depolarization of Vbl in control tubules and a 33.0 +/- 4.2 mV depolarization in tubules from DOCA-pretreated rabbits. Concurrently, Ra/Rbl decreased in both groups, consistent with the existence of a conductive K+ channel at the apical cell membrane. This apical K+ channel was not sensitive to amiloride but was blocked by Ba2+. Conductive movement of Na+ across the apical membrane was also apparent in that Ra/Rbl increased with amiloride from 0.61 +/- 0.10 to 1.45 +/- 0.28. A 10-fold increase in the bath [K+] caused a 28.6 +/- 3.8 and a 49.4 +/- 4.4 mV depolarization of Vbl in tubules obtained from control and DOCA-pretreated rabbits, respectively. In both groups Ra/Rbl increased, suggesting that the basolateral cell membrane also contains a conductive K+ channel. Taken together the results support a model in which the transepithelial reabsorption of Na+ and the transepithelial secretion of K+ are driven by the Na+ K+-ATPase located in the basolateral cell membrane, with passive movement of these ions occurring through separate conductive pathways in the apical cell membrane. PMID- 6295186 TI - Enterocyte alpha 2-adrenergic receptors: yohimbine and p-aminoclonidine binding relative to ion transport. AB - We previously reported that alpha 2-adrenergic agonists enhance absorption and inhibit secretion of electrolytes in small intestine. The present study was undertaken to characterize and localize the relevant receptors. Plasma membranes derived from isolated rabbit ileal epithelial cells were incubated with either [3H]yohimbine (Yo), an alpha 2-antagonist, or p-[3H]aminoclonidine (PAC), an alpha 2-agonist. Scatchard analysis of [3H]Yo binding suggests a single receptor. Competitive displacement of Yo from this receptor by other ligands had a potency order characteristic for alpha 2-receptors in other tissue systems. A Scatchard plot of [3H]PAC binding was curvilinear and best fit by assuming two independent site. Competitive displacement of [3H]PAC by PAC in the presence of 140 mM Na+ or 0.1 mM GTP increased the IC50 for PAC binding from 10 nM to 100 and 105 nM, respectively, and the Hill coefficient from 0.7 to 1.2 and 1.0, respectively. The ED50 for PAC effect on short-circuit current (200 nM) does not differ significantly from these values. We conclude that alpha 2-receptors are present on ileal enterocytes and that these receptors mediate enterocyte fluid and electrolyte transport function. PMID- 6295185 TI - Anion effects on fluid absorption from rat jejunum perfused in vivo. AB - Perfusion of rat jejunal segments in vivo with an isotonic, HCO3-free SO4-Ringer solution resulted in low rates of net sodium (JNanet) and water absorption. When the perfusion fluid was changed to one containing 25 mM Na2SO3, JNanet increased from 4.7 +/- 1.2 to 11.6 +/- 1.5 (SE) mumol X cm-1 X h-1 (P less than 0.001). This increased absorption was accompanied by comparable increases in chloride and water absorption, occurred without a detectable change in potential difference across the perfused segment, and was readily reversed on reinstitution of perfusion with SO4-Ringer. Perfusion with SO3-Ringer had no effect on electrolyte absorption from terminal segments of rat ileum. Addition of L-phenylalanine stimulated absorption from SO4-Ringer perfusate but not from SO3-Ringer perfusate. Addition of 25 mM NaHCO3 to SO4-Ringer perfusate caused parallel increases in JNanet and JHCO3net; when 25 mM NaHCO3 was added to SO4-Ringer perfusate that also contained 25 mM NaSCN, the same increase in JHCO3net occurred but was not associated with any increase in JNanet. These results indicate a potent effect of SO2-3 and HCO-3 to stimulate JNanet from rat jejunum but not from ileum. These anion effects on intestinal transport in vivo resemble their effects on ATPase activity of brush-border fractions from small intestine in vitro and raise the possibility that these effects on ion transport could be mediated through the changes in brush-border ATPase activity, which are brought about by exposure to these anions, although other explanations are also possible. PMID- 6295187 TI - Maximum fluid concentrations of materials released from platelets at a surface. AB - We examine the estimation of local concentrations of materials that are released from the dense and alpha-granules of platelets during accumulation of platelets upon collagen-coated glass. Platelet/red blood cell suspensions were perfused through a 1.3-mm-ID tube. Empirical data were used in a calculation procedure, based on diffusion and convection, designed to yield an upper bound on the interfacial fluid concentration (IFC) for each substance considered. The necessary empirical data are the rate of platelet accumulation and the maximum amount of material in the platelet capable of secretion. It was found that the IFC is dependent on the shear rate at the surface (G) and is proportional to G0.27. This means that an eightfold increase in flow rate would increase the IFCs approximately twofold. Serotonin, pyrophosphate, adenosine 5'-monophosphate (AMP), and adenosine 5'-triphosphate (ATP) were found not to be present in sufficient quantities to produce IFCs that could influence platelet aggregation if used alone at the IFC. A second set of materials, fibrinogen, fibronectin von Willebrand factor, and calcium, had IFCs less than their concentrations normally found in plasma. A third category, containing adenosine 5'-diphosphate (ADP) alone, had an IFC close to those known to affect platelet aggregation. The role of metabolites of arachidonic acid, which may promote or inhibit platelet aggregation, awaits further description. PMID- 6295188 TI - A mathematical model of the vagally driven primary pacemaker. AB - Information regarding the electrical behavior of cells in the sinoatrial (SA) node of the heart is by no means complete; yet sufficiently detailed information is available in the literature to allow the formulation of a reasonably quantitative model of the membrane of the primary pacemaker cell (P-cell) and its parasympathetic innervation. In a previous study [Am. J. Physiol. 243 (Heart Circ. Physiol. 12): H207-H218, 1982], the well-known McAllister-Noble-Tsien model of the cardiac Purkinje fiber is modified to account for the electrical activity of the SA node. In the present study, the effects of vagal activity on sinus rhythm are considered by adding a special acetylcholine-sensitive "muscarinic channel" to the membrane model for the P-cell. The resulting model mimics published data quite well and is capable of characterizing the free-running SA node cell as well as its responses to electrotonic and vagal stimulation. PMID- 6295189 TI - beta-Adrenergic activity and cardiovascular response to severe respiratory acidosis. AB - The mechanism responsible for the depressive myocardial effects of severe respiratory acidosis is unclear; however, sympathetic stimulation and catecholamines are known to be involved. The influence of beta-adrenergic receptor activity on the myocardial response to severe respiratory acidosis was studied in 18 anesthetized, mechanically ventilated dogs. Arterial CO2 tension (PaCO2) was raised by increasing the inspired CO2 fraction in O2. In control animals, as PaCO2 increased, heart rate (HR) decreased (PaCO2 approximately 110 mmHg), then returned to control (PaCO2 approximately 220 mmHg), whereas arterial blood pressure (Pa) and cardiac output (Q) remained unchanged from prehypercapnia levels. At PaCO2 greater than 350 mmHg, Pa, HR, and Q decreased and left ventricular function (LVF) curves were depressed. Death occurred at a PaCO2 of 404 +/- 25 mmHg (pH 6.48 +/- 0.02). In a second group of animals, administration of isoproterenol during the increase in PaCO2 did not result in depression of myocardial function, and death did not occur even at a significantly higher PaCO2 (PaCO2 496 +/- 12 mmHg; pH 6.39 +/- 0.02) than in the control group. Administration of propranolol to a third group of animals as PaCO2 increased did not change Pa, HR, and Q; however, LVF curves indicated a more rapid and severe depression of myocardial performance than in control, and death occurred at a significantly lower PaCO2 (PaCO2 220 +/- 25 mmHg; pH 6.65 +/- 0.02). We conclude that beta-adrenergic receptor stimulation can prevent hypercapnic heart failure and that beta-adrenergic receptor activity is involved in the mechanism responsible for this failure. PMID- 6295190 TI - Regression of myocardial hypertrophy and influence of adrenergic system. AB - Studies of regression of myocardial hypertrophy in spontaneously hypertensive rats (SHR) suggest that the adrenergic system may play an important role in the reversal of hypertrophy. The effect of propranolol on reversal of hypertrophy, however, is still controversial. This study describes the effect of propranolol, given alone or in combination with hydralazine in different ratios for 4 wk, on blood pressure (BP), ventricular weight, and myocardial catecholamine (MC) concentrations. The data show that a certain ratio of propranolol to hydralazine (750:30) leads to moderate BP control (196-156 mmHg) without increased MC (634 vs. 552 ng/g) and moderately reduced hypertrophy. Reduction of BP alone with increased MC (hydralazine alone) or reduction of MC without BP control (propranolol alone) failed to reduce hypertrophy. A significant correlation between both ventricular weight and heart rate with MC (r = 0.6) was obtained by multiple regression analysis. This study suggests that adrenergic factors seem to play an important role in modulating structural cardiac response to variations in arterial pressure. PMID- 6295191 TI - Incomplete free fatty acid oxidation by ascites tumor cells under low oxygen tension. AB - We tried to understand why our earlier estimates of fatty acid (FA) oxidation rates under the nearly anaerobic state of the Ehrlich ascites tumor (EAT) in vivo were even greater than those found in vitro under aerobic conditions. Using tracers [1-14C]linoleate, [1-14C]-, and [9,10-3H]palmitate, and NaH14CO3, we estimated essential and nonessential FA oxidation rates to CO2 + H2O by EAT in living mice and in vitro under aerobic and anaerobic conditions. Sequestration of intraperitoneally (ip)-injected 14C-FFA allowed a selective labeling of the tumor versus the host; thus, breath 14CO2 could be used to estimate the maximum rate of FA oxidation in vivo by the tumor. Initially, we measured breath 14CO2 following NaH14CO3 injections and developed a multicompartmental model to simulate the tumor-host HCO-3-CO2 system. This model was integrated with our earlier model for tumor FA turnover. The integrated model was fitted to breath 14CO2 data from mice injected ip with 14C-FFA to compute tumor FA oxidation rates. Both essential and nonessential FA were oxidized to CO2 at similar rates. The maximum rate of total FA oxidation to CO2 was 5-6 nmol FA X min-1 X 7-ml tumor-1, about 5-10 times lower than all previous estimates obtained in vitro and in vivo. To resolve this dilemma we used doubly labeled [1-14C; 9,10-3H]palmitate and found that under aerobic conditions, in vitro, EAT formed 3H2O and 14CO2 at nearly equal rates. These rates were suppressed markedly but unequally at low PO2. Anaerobic suppression of 14CO2 formation greatly exceeded that of 3H2O formation. As a result 3H2O/14CO2 reached a value of congruent to 10 at low PO2. Our data indicate that under the nearly anaerobic conditions of a growing EAT in vivo, the partial beta-oxidation of FA to 2C + H2O takes place at a 5 to 10 times faster rate than the complete oxidation of FA to CO2 + H2O. This finding can account for earlier apparent inconsistencies in the literature, since aerobic studies of 14C FA oxidation to 14CO2 in vitro and of 3H-FA oxidation to 3H2O under nearly anaerobic conditions would both overestimate greatly the rate of FA oxidation to CO2 by EAT in vivo. PMID- 6295192 TI - Adverse interaction between disulfiram and marijuana: a case report. AB - The authors describe a hypomanic-like reaction in a patient taking disulfiram and marijuana simultaneously and suggest there was an adverse interaction between the two drugs. PMID- 6295193 TI - Hepatoblastoma. Attempt at characterization of histologic subtypes. AB - This is a clinicopathologic study summarizing the experience with hepatoblastomas at Children's Memorial Hospital of Chicago between 1954 and 1981. Of 21 patients studied, 13 (61.9%) died. Three major histologic epithelial patterns were identified: fetal, embryonal, and macrotrabecular. The first two may represent different stages of cytodifferentiation of hepatoblastoma cells. The macrotrabecular type had features similar to hepatocellular carcinoma of adults, from which distinction was difficult; this type pursued an aggressive clinical course. The fetal type exhibited advanced differentiation, and two cases in this category survived after surgery only; local extrahepatic dissemination was present in other cases of the fetal type. Mixed epithelial and mesenchymal tumors constituted only 23% of this series, and none contained rhabdomyoblastic elements. Although modern chemotherapy may alter the course of this disease, the small size of this series precluded definite statements in this regard. Only patients in whom the tumor was completely excised as primary treatment became long-term survivors. PMID- 6295194 TI - Primary thymic carcinomas. AB - Within a 75-year period, 20 patients with primary nonteratomatous carcinomas of the thymus were seen at the Mayo Clinic. Fourteen were males and six were females, with a mean age of 48 years. Thirteen of the lesions were classified as poorly differentiated or spindling squamous cell carcinomas. One neoplasm was a sarcomatoid carcinoma, and two showed both squamous cell and neuroendocrine ultrastructural features. The remaining four tumors had both light- and electron microscopic features of neuroendocrine carcinomas. In all 20 patients, careful clinical and pathologic examinations, including autopsy in fatal cases, excluded malignant lymphomas and metastasis to the thymus from epithelial tumors in other anatomic sites. Thirteen patients underwent surgery, with or without postoperative radiotherapy; of these, one was alive and apparently disease-free 43 months after diagnosis. There were two operative deaths. The remaining patients received primary radiotherapy or chemotherapy, or both. The mean survival of the 16 patients who died (excluding operative deaths) was 18.7 months for the 14 patients with tumors of squamous cell differentiation and 36.0 months for the two patients with pure neuroendocrine carcinomas. Among patients with squamous cell carcinoma, 11 developed metastases to lungs, liver, bones, adrenal glands, or extrathoracic lymph nodes; nine of these died of massive local growth within the mediastinum. Among patients with neuroendocrine carcinomas, two of four patients died of local growth within the mediastinum; one of these also had metastases to the liver and adrenal glands. One patient with oat cell carcinoma was alive with residual thoracic tumor and cervical lymph node metastasis 18 months after diagnosis. PMID- 6295195 TI - Malignant granular cell tumor. Report of a case with special reference to carcinoembryonic antigen. AB - A case of malignant granular cell tumor and its histochemical and electron microscopic characteristics are reported. This case showed, in addition to the well-known distribution of this type of tumor in subcutaneous fat, mediastinum, retroperitoneum and lungs, multiple foci in the myocardium. Contrary to recent studies reporting the presence of carcinoembryonic antigen (CEA) in benign and malignant granular cell tumors, this case is CEA-negative. We suggest that the reported CEA-reactivity in this type of tumor is probably due to cross-reacting antibodies against antigens, presumably associated with lysosomes. PMID- 6295196 TI - Caroli's Disease: a premalignant condition? AB - This study reports four new cases of Caroli's disease complicated by the development of cholangiocarcinoma. Caroli's disease appears to be a premalignant condition, but early diagnosis of malignancy in the cyst wall is difficult because signs of ductal stenosis develop slowly and treatment is often based on erroneous diagnosis of recurrent cholangitis. When carcinoma is present, operative intervention makes no difference in survival. In patients with Caroli's disease, adequate intraductal and hepatic biopsies should be obtained at the first operation. These patients should be followed carefully, and malignant growth should be suspected when cholangitis occurs despite patent anastomoses. Earlier diagnosis will obviate repeated operations in patients with malignant Caroli's disease. PMID- 6295199 TI - Effect of 4-aminopyridine on neurotransmission in smooth muscle. PMID- 6295197 TI - Increased incidence of experimental colon cancer associated with long-term metronidazole therapy. AB - Using the well-established DMH model for colon neoplasia, we demonstrated that a high-fiber diet pair-fed to animals was associated both with certain changes in bacterial profile and with protection against experimental colon neoplasia. The addition of metronidazole on a long-term basis to both high- and low-fiber diets did not alter stool bacteroides counts as expected and was associated with an apparent cocarcinogenic effect. Concern exists among surgeons and gastroenterologists as to whether metronidazole places their patients at risk. The status of long-term metronidazole therapy for patients with Crohn's disease is a pertinent example. In view of our findings, it is important to further elucidate the metabolism of metronidazole in both the rat and human gut. PMID- 6295198 TI - Serum neuron-specific enolase: a serum marker for nonfunctioning pancreatic islet cell carcinoma. AB - The isoenzyme, neuron-specific enolase (NSE) was evaluated as a serum marker for nonfunctioning pancreatic islet cell carcinoma. Serum NSE was measured by radioimmunoassay in 6 patients with islet cell cancer and in 22 healthy adults. Mean serum NSE in control subjects was 5 ng/ml (range 3.2 to 8.4 ng/ml). Three of six patients had clearly elevated serum NSE with values that ranged from 22 to 44 ng/ml. One patient had an equivocally elevated serum NSE of 9.2 ng/ml while receiving chemotherapy. When relapse occurred, the NSE level increased to 44 ng/ml. One of two patients with normal serum NSE had undergone radical pancreatectomy and had no evidence of disease when studied. Elevated serum NSE returned to normal limits after removal of all gross tumor in the only patient from whom a preoperative measurement was obtained. Results of this preliminary study suggest that serum NSE levels can aid in the diagnosis and monitoring of the course of nonfunctining islet cell carcinoma. PMID- 6295200 TI - [Characteristics of various adaptive reactions in patients with endometriosis]. PMID- 6295201 TI - [Ovarian and adrenal function in women with hyperandrogeny of different etiology]. PMID- 6295202 TI - [Cytomegalovirus infections during pregnancy]. PMID- 6295203 TI - The determination of acetaldehyde in human blood by the perchloric acid precipitation method: the characterization and elimination of artefactual acetaldehyde formation. PMID- 6295204 TI - Purification of radiolabeled and native polypeptides by gel permeation high performance liquid chromatography. PMID- 6295205 TI - Rapid recording luminometric assay of adenosine 3',5'-cyclic monophosphate phosphodiesterase activity and a possible pitfall in coupled enzyme assays. PMID- 6295206 TI - Development of competitive enzyme immunoassays for human serum angiotensin-1 converting enzyme: a comparison of four assay configurations. PMID- 6295207 TI - Separation of basic, hydrophilic peptides by reversed-phase ion-pair chromatography. II. Analytical applications with particular reference to phosphoserine peptides. PMID- 6295208 TI - Periodate oxidation of methionine in proteins. PMID- 6295209 TI - An improved method for preparing large arrays of bacterial colonies containing plasmids for hybridization: in situ purification and stable binding of DNA on paper filters. PMID- 6295210 TI - A method for isolation of a large amount of a single-stranded DNA fragment. PMID- 6295211 TI - Quantitative visible spectroscopy at low temperatures: a systematic examination. PMID- 6295212 TI - High-performance liquid chromatographic determination of vitamin D3 in foods with particular reference to eggs. PMID- 6295213 TI - Safety of nerve transmission. PMID- 6295215 TI - The clinical use of glucocorticoids. AB - Glucocorticoids are potent anti-inflammatory agents and their administration results in a wide range of effects on inflammatory and immunologically mediated disease processes. The precise mechanisms by which glucocorticoids impair the human immune response are unknown. Intracytoplasmic glucocorticoids specific receptors are important in the specificity of glucocorticoid actions. Glucocorticoid administration results in neutrophillia, monocytopenia, lymphopenia, and eosinopenia. A principle mechanism whereby glucocorticoids limit inflammation is by limiting the access of leukocytes, particularly neutrophils, to inflammatory sites. Neutrophil function is relatively refractory while monocyte and T-cell function is more easily impaired. A variety of glucocorticoid preparations are available for use, and appreciation of their relative potency and plasma half-lives is essential for designing therapeutic regimens. High doses and frequent administration of glucocorticoids are necessary in order to induce a remission in patients with flagrantly active disease. Once a remission is induced, the glucocorticoid regimen should be adjusted to attain maximal therapeutic benefit with minimal adverse effects. Alternate day dosage regimens can often be used to maintain a remission. PMID- 6295214 TI - The inactivation of the bovine J blood group substance by the periodate ion. AB - 1. Treatment of J-positive (Jcs) bovine erythrocytes with periodate (0.25 mmol/l final concentration, 1 hour, room temperature) has no effect on the J activity. Higher periodate concentrations cause spontaneous haemolyses. 2. Treatment of the lipids extracted from (and containing all J activity of) Jcs erythrocytes with periodate leads to a decrease of J activity even with lower periodate concentrations. 3. Treatment of the stroma prepared from Jcs erythrocytes with periodate demonstrated the relative stability of the J antigen up to 0.25 mmol/l periodate. At the same time the sialic acid concentration of stroma is reduced to about 13% of the initial concentration. 4. Desialylation of Jcs erythrocytes or Jcs stroma with sialidase does not affect the J activity thus confirming previous findings. On the other hand, the J activity of desialylated Jcs stroma is much more susceptible to periodate. 5. It is concluded that membrane-bound sialic acid shields the membrane-bound J antigen from being attacked by periodate. PMID- 6295216 TI - [Cytomegalovirus infection in general surgery. Two cases]. PMID- 6295217 TI - [Psychomotor hyperexcitability induced by fentanyl in anesthesia]. PMID- 6295218 TI - Pharmacology and clinical use of neuromuscular blocking agents. PMID- 6295219 TI - Interaction of bluetongue virus with preimplantation embryos from mice and cattle. AB - Preimplantation embryos from mice and cattle were exposed to bluetongue virus in vitro to determine whether the virus would replicate in these early embryos and, if so, what pathologic consequences would ensue. A high proportion of zona pellucida-free, 2-cell embryos and morulae from mice, and morulae from cattle became infected. The infection was rapidly cytopathic in embryos from both species. Indirect immunofluorescence was used to demonstrate accumulation of virus antigen in the blastomeres of these embryos. The zona pellucida of both murine and bovine embryos provided effective protection from virus present in culture fluid. PMID- 6295220 TI - Responses of serum and lung angiotensin-converting enzyme activities in the early phase of pulmonary damage induced by oleic acid in dogs. AB - Changes in angiotensin-converting enzyme (ACE) activity in the lung tissue, edema fluid, and bloodstream were studied during induction of lung edema by oleic acid. Left lungs of 25 mongrel dogs were unilaterally treated with oleic acid (0.1 ml/kg) injected into the right atrium during right pulmonary artery occlusion for 2.5 min. Lung tissue and serum ACE activities and physiologic parameters were followed for as long as 180 min. Serum ACE activity increased to 106% at 2.5 min (p less than 0.025) and to 128% at 180 min. The ACE activity of treated lung tissue decreased compared with the zero time control in terms of tissue DNA content (at 180 min, p less than 0.05). Furthermore, the left to right activity ratio of the precipitate fraction decreased to 0.59 as early as 5 min after oleic acid injection (p less than 0.01). Changes in these ACE activities preceded those of arterial oxygen tension and base transthoracic electrical impedance. Edema fluid at 45 min had a specific activity that was 1.62 times greater than that in serum. These data indicated that the change in ACE activity reflected the impairment of pulmonary vascular endothelial cells and that the oleic acid injured endothelial cells of the pulmonary capillaries as early as 2.5 min after administration. PMID- 6295221 TI - Treatment of mixed-dust pneumoconiosis with whole lung lavage. AB - A patient with silicosis and progressive dyspnea on exertion is described in whom open lung biopsy revealed active chronic inflammation with many macrophages filling alveolar spaces. Because of the extensive involvement by the disease of small air spaces, bilateral whole lung lavage was performed. The lung lavage effluent was striking in its blackish brown color. It was composed predominantly of macrophages containing silica, silicates, and graphite. Particles in the tissue and lavage were analyzed using scanning electron microscopy and energy dispersive X-ray analysis. The dry weight of the material removed was approximately 25 g, of which an estimated 135 mg was silica. The procedure resulted in immediate symptomatic improvement in the patient. Although his pulmonary function did not change significantly, it is hoped that removal of this material will improve his long-term prognosis. PMID- 6295222 TI - Alterations in leukocyte oxidative metabolism in cigarette smokers. AB - The polymorphonuclear leukocyte (PMN) may play an important role in the pathogenesis of lung disease associated with cigarette smoking. To investigate its potential for oxidant-mediated lung injury in cigarette smokers, we studied PMN oxidative metabolism in asymptomatic cigarette smokers and nonsmoking control subjects. We found a marked increase in oxidant release in a group of cigarette smokers. After stimulation by phorbol myristate acetate, release of superoxide anion (O-2) by PMN in smokers with white blood counts (WBC) greater than 9,000 was 50% greater than in nonsmokers with similar WBC or smokers and nonsmokers with WBC less than 9,000. Abstinence from smoking did not affect the alterations in O-2 release nor did a serum factor appear responsible. The changes appeared to be part of a generalized increase in oxidative metabolism, as there was greater oxidation of glucose (1-14C) and chemiluminescence by PMN from smokers with WBC greater than 9,000. A further estimate of lung oxidant load was determined by evaluating the marginated pool of PMN. Smokers with WBC greater than 9,000 showed a 70% increase in WBC after epinephrine, and PMN oxidative metabolism remained increased in this group. This study demonstrates that cigarette smokers with elevated WBC have increased release of potentially toxic oxygen metabolites. These cigarette smokers also demonstrated increased oxidant release from the marginated PMN pool. Because leukocyte-generated oxygen metabolites are highly reactive and can cause tissue injury, these findings may have important implications in the pathogenesis of smoking-related lung disease. PMID- 6295223 TI - The value of bronchoscopy in establishing the etiology of pneumonia in renal transplant recipients. AB - Fifty-two episodes of fever and new pulmonary infiltrates were evaluated prospectively in 51 renal allograft recipients. Thirty-nine flexible fiberoptic bronchoscopies were performed in the diagnostic evaluation of these infiltrates. Specific etiologic diagnoses were obtained in 30 (77%) of the patients. This information was clinically useful, as defined by preset criteria, in 21 (54%) of the patients and definitive but not clinically useful in an additional 9 (23%). In the remaining 9, it was neither definitive nor clinically useful. Microbiology brush specimens were useful in establishing etiologic diagnoses in 12 (44%) of the 27 patients in whom it was performed. Transbronchial lung biopsies yielded specific etiologic diagnoses in 9 (53%) of the 17 biopsies obtained. Complications related to the bronchoscopic procedure occurred in 2 patients (5% of total bronchoscopies). No prolonged morbidity was noted. We conclude that fiberoptic bronchoscopy is a safe, useful procedure, and should be considered early in the diagnostic evaluation of pulmonary infections in renal transplant recipients. PMID- 6295224 TI - Interstitial collagenase secretion and giant cell formation from rabbit alveolar macrophages. Effects of dexamethasone. AB - Rabbit alveolar macrophages (AM) induced by complete Freund's adjuvant have been used as a model for activated AM. We studied the effects of in vivo and in vitro corticosteroid on collagenase release and spontaneous formation of multinucleated giant cells (MGC) in culture with this system. Dexamethasone 10- through 10(-7) molar concentration in vitro inhibited both collagenase release and spontaneous AM fusion. Daily in vivo administration of dexamethasone to as much as 0.128 mg/kg similarly suppressed these AM functions in culture. These studies show that in vitro and in vivo corticosteroids inhibit collagenase release and MGC formation of rabbit AM in culture in doses comparable to those used therapeutically in humans. This model may be useful in examining the mechanisms of cell fusion and functions of MGC in granulomatous lung disease. PMID- 6295226 TI - Exaggerated natriuretic response of Brattleboro rats to extracellular volume expansion. PMID- 6295225 TI - Endothelial cell damage and tubuloreticular structures in interstitial lung disease associated with collagen vascular disease and viral pneumonia. AB - Lung tissue showing the light microscopic changes of interstitial inflammation and fibrosis was studied electron-microscopically in 11 patients with collagen vascular disease and 26 patients with viral pneumonia. All patients exhibited progressive respiratory insufficiency and had abnormal pulmonary function test results. Endothelial cell cytoplasmic swelling and intracellular tubuloreticular structures were the dominant ultrastructural change in the lungs of both groups. Tubuloreticular structures were identified in peripheral blood lymphocytes in all patients with collagen vascular disease and in 2 with viral pneumonia. Tubuloreticular structures were not identified in the lung tissue from various control patients and in the lymphocytes from patients with idiopathic interstitial pulmonary fibrosis. We conclude that (1) the ultrastructural changes seen in viral pneumonia and collagen vascular-associated interstitial lung disease are essentially identical; (2) endothelial cell damage may be the primary pathologic change associated with both diseases; (3) tubuloreticular structures are seen in large numbers in lung tissue from patients with viral pneumonia and collagen vascular disease; (4) tubuloreticular structures are present in peripheral blood lymphocytes from all patients with collagen vascular disease. PMID- 6295227 TI - ACTH and indomethacin-induced antidiuresis in low salt Brattleboro strain rats: induction and escape. PMID- 6295228 TI - The renin-angiotensin system in the absence of antidiuretic hormone. PMID- 6295229 TI - Effect of lithium and antidiuretic hormone on plasma renin concentration in diabetes insipidus rats (Brattleboro rat model). PMID- 6295230 TI - Use of the Brattleboro rat in studies of the mechanism by which chlorpropamide enhances the action of vasopressin. PMID- 6295232 TI - Fetal and postnatal maturation of corticotrope function in the vasopressin deficient rat (Brattleboro strain): a radioimmunological, immunocytochemical, and morphometric study. PMID- 6295233 TI - Molecular forms and anterior pituitary content of ACTH, beta-lipotropin, and beta endorphin in diabetes insipidus di/di (Brattleboro) rats. PMID- 6295231 TI - The hormonal status of the Brattleboro rat. PMID- 6295234 TI - Potentiation of hypothalamic corticotropin releasing activity by vasopressin: studies in the Brattleboro rat. PMID- 6295235 TI - Central control of ACTH secretion in diabetes insipidus Brattleboro rats. PMID- 6295237 TI - Oxytocin action in isolated adipocytes from Brattleboro rats. PMID- 6295236 TI - Role of vasopressin in the renin and ACTH responses to intraventricular angiotensin II. PMID- 6295238 TI - Influence of posterior pituitary hormones on the pituitary-adrenocortical response to neurogenic stress in the Brattleboro rat. PMID- 6295239 TI - The hypothalamic median eminence of the homozygous Brattleboro rat is deficient in calcitonin-specific binding sites. PMID- 6295240 TI - Immunological studies of Down's syndrome and Alzheimer's disease. PMID- 6295241 TI - Long-term inhalation studies with raw and processed shale dusts. PMID- 6295242 TI - Translocation of mineral fibres through the respiratory system after injection into the pleural cavity of rats. PMID- 6295243 TI - The dust content of the lungs of hard-rock miners and its relationship to occupational exposure, pathological and radiological findings. PMID- 6295244 TI - Asbestos fibres in the lungs of chrysotile miners and millers--a preliminary report. PMID- 6295245 TI - Mineral fibre content of lung in mesothelial tumours in North America. PMID- 6295247 TI - Variations in the carcinogenicity of mineral fibres. PMID- 6295246 TI - Comparative cocarcinogenic effects of crocidolite asbestos, hematite, kaolin and carbon in implanted tracheal organ cultures. PMID- 6295248 TI - Effect of impurities and associated minerals on quartz toxicity. PMID- 6295249 TI - Estimates of dose-response for respiratory cancer among chrysotile asbestos textile workers. PMID- 6295250 TI - Radiological changes and fibre exposure in chrysotile workers aged 60-69 years at Thetford Mines. PMID- 6295251 TI - A mortality study of workers manufacturing friction materials with chrysotile asbestos. PMID- 6295252 TI - Cell-mediated immunity in herpetic keratitis measured by leukocyte migration inhibition. AB - Leukocyte migration inhibition factor (LMIF) assays have been used as a measure of cell-mediated immune responses. Direct assays of this factor were determined in 30 patients with recurrent herpes simplex stromal keratitis during the quiescent stage of the disease and in 10 of these patients during the acute stage as well. The migration of leukocytes incubated in the presence of HSV antigens was compared to that without viral antigens for the migration index (MI). Only 1 out of 30 patients had a positive response during the quiescent stage, while among the 10 patients with the active disease, four had a positive response. PMID- 6295253 TI - [Klippel-Trenaunay syndrome with systemic hypertension and chronic renal failure]. PMID- 6295254 TI - Granular cell tumor. PMID- 6295256 TI - Neonatal-onset adrenoleukodystrophy in a girl. AB - A 4-year 11-month-old girl developed cerebral degeneration with onset in the neonatal period. Postmortem examination showed gross, microscopic, ultrastructural, and biochemical changes identical to those associated with adrenoleukodystrophy (ALD), a sex-linked disorder of boys beginning in the first decade of life. Cytoplasmic inclusions ultrastructurally identical to those in brains and adrenal glands of ALD patients were present not only in this girl's adrenal glands and brain but also in reticuloendothelial cells of the liver, lymph node, spleen, thymus, and hepatic lysosomes. Thin-layer and gas-liquid chromatographic analysis of cerebral tissues demonstrated abnormal long-chain fatty acids in the cholesterol ester fraction, identical to those present in affected tissues of males with ALD. The documentation of abnormal long-chain fatty acids in cerebral tissues of a female patient supports the concept that infantile-onset ALD is a clinically and pathologically distinctive entity characterized by prominent visceral storage abnormalities and occurrence in both sexes. These findings also suggest that ALD and related entities are either phenotypic variants of a specific enzyme deficiency or separate disease resulting from different mutations in a common metabolic pathway. PMID- 6295257 TI - [Trial use of polymyxin and erythromycin for isolating cultures of the plague microbe in the Kyzyl-Kum]. AB - It is recommended that the Hottinger agar medium with polymyxin or erythromycin be used for isolation of pure cultures of the plague microbe from specimens containing various species of microorganisms, except Proteus. Such a procedure was used in the Kizil-Kum for the bacteriological examinations with respect to plague. The procedure may be used in the natural foci with the strains resistant to these antibiotics. PMID- 6295255 TI - Effects of diethyldithiocarbamate on organ distribution and excretion of cadmium. AB - Sodium diethyldithiocarbamate (DDTC) was evaluated for its efficacy in promoting organ mobilization and excretion of metallothionein-bound cadmium (Cd) using mice which received a single injection of CdCl2 X 2.5 H2O, 0.03 mg per mouse, along with 109CdCl2 three to six weeks earlier. After seven or 13 i.p. injections of DDTC, 500 mg per kg, over a two or four week interval, DDTC was highly effective in mobilizing Cd from kidney (Ki) and spleen (Sp), but less effective in removing it from liver (Li). Treatment with DDTC moderately enhanced Cd levels in lung (Lu), heart (He) and testes (Te), and increased brain (Br) levels to over 500 percent of control values. Relative accumulation of Cd in organs of control mice were in the order Li greater than Ki greater than Sp greater than Lu greater than He greater than Te greater than Br. The extreme values were Li = 57 percent and Br = 0.07 percent of the Cd administered. Even though a major portion of Cd mobilized was from the kidneys, excretion was apparently exclusively by the fecal route. PMID- 6295258 TI - [Changes in the biological properties of the pertussis microbe exposed to antibiotics]. AB - Experimental modeling of the process of variation of the pertussis causative agent grown on the casein-carbon agar medium with subtoxic doses of the widely used antibiotics was performed. Significant changes in the morphological, cultural and serological properties of the pertussis microbe, in its sensitivity to the antibiotics and bacteriocins, in the activity of glutamine synthetase and in the electrophoretic mobility of the protein components were shown. The antigen structure determined with the method of immune electrophoresis in the agar gel and the biochemical properties were the stable characteristics defining the population taxonomic position as Bordetella pertussis. Possible occurrence of atypical strains of the pertussis causative agent in the patients with the cough syndrome treated with the antibiotics is indicated. It is suggested that antibiotics may play a significant role in redistribution of subpopulations with different functional properties in the population of the pertussis microbe. PMID- 6295259 TI - [Ultrastructural changes in populations of the pertussis microbe exposed to antibiotics]. AB - The ultrastructural changes in the populations of the pertussis causative agent maintained for a prolonged period of time on nutrient media containing antibiotics were studied with the method of electron microscopy. Significant changes in the ultrastructure of the cells and the population composition due to the effect of tetracycline, levomycetin or erythromycin were observed. The ultrastructure of the populations affected by penicillin or streptomycin and their biological properties were most similar to those of the initial strains. The electronogrammes revealed morphological heterogeneity of the initial and experimental cultures and changes in the subpopulation composition induced by the antibiotics. PMID- 6295260 TI - Antipicornavirus flavone Ro 09-0179. AB - Ro 09-0179 (4',5-dihydroxy-3,3',7-trimethoxyflavone), isolated from a Chinese medicinal herb, was found to have potent antiviral activity. It selectively inhibited the replication of human picornaviruses, such as rhinoviruses and coxsackieviruses in tissue culture, but not other DNA and RNA viruses. Ro 09-0298 (4',5-diacetyloxy-3,3',7-trimethoxyflavone), an orally active derivative of Ro 09 0179, prevented coxsackievirus (B1) infection in mice. The critical time for the inhibition of rhinovirus replication by Ro 09-0179 was 2 to 4 h after virus adsorption, i.e., in the early stages of virus replication. It markedly inhibited coxsackievirus and rhinovirus RNA synthesis in infected HeLa cells, but not in a cell-free system using the RNA polymerase complex isolated from the infected cells. In the infected cells, the RNA polymerase complex was not formed in the presence of Ro 09-0179. Therefore, it is suggested that Ro 09-0179 interferes with some process of viral replication which occurs between viral uncoating and the initiation of viral RNA synthesis. PMID- 6295261 TI - Direct and specific inactivation of rhinovirus by chalcone Ro 09-0410. AB - Studies of various analogs related to the antipicornavirus agent, 4',5-dihydroxy 3,3',7-trimethoxyflavone (Ro 09-0179), led to the identification of 4'-ethoxy-2' hydroxy-4,6'-dimethoxychalcone (Ro 09-0410), a new and different type of antiviral agent. Ro 09-0410 had a high activity against rhinoviruses but no activity against other picornaviruses. Of 53 rhinovirus serotypes so far tested, 46 were susceptible to Ro 09-0410 in HeLa cell cultures. The concentration of Ro 09-0410 inhibiting 50% of the types of rhinovirus was about 0.03 micrograms/ml, whereas the 50% cytotoxic concentration was 30 microgram/ml. Ro 09-0410 inactivated rhinoviruses in direct dose-, time-, and temperature-dependent fashion. Since infectivity, reduced by exposure to the agent, completely regained the original level by extraction of the agent with chloroform, the inactivation may be associated with the binding of the agent to some specific site of the rhinovirus capsid. PMID- 6295262 TI - Pharmacokinetics and cerebrospinal fluid bactericidal activity of ceftriaxone in the treatment of pediatric patients with bacterial meningitis. AB - Single-dose pharmacokinetics of ceftriaxone were determined in 19 patients with proven bacterial meningitis. The dosage was 50 mg of ceftriaxone per kg. The plasma concentration time curve declined in a biexponential manner. The mean peak plasma concentration was 207 micrograms/ml, and the elimination half-life was 4 h. In 12 patients, multiple-dose pharmacokinetics were determined after a loading dose of 75 mg of ceftriaxone per kg, followed by 50-mg/kg doses every 8 h in 5 patients or every 12 h in 7 patients. The mean peak plasma concentration was 230 micrograms/ml after the first dose and 263 micrograms/ml after the last dose. Of 12 patients, 5 had trough values that were larger after multiple doses than after a single dose. Mean penetration of ceftriaxone into cerebrospinal fluid was 3.1%. The median cerebrospinal fluid bactericidal titer against the patients pathogens was greater than 1:1,024 and less than 1:2,048. The drug was well tolerated without adverse effects. PMID- 6295264 TI - In vitro activity of cefodizime (HR-221). AB - The in vitro activity of cefodizime (HR-221), a new cephalosporin antibiotic, was compared with the activities of selected antimicrobial agents against a broad spectrum of aerobic bacteria. Cefodizime concentrations of 2 micrograms/ml inhibited about 90% of Enterobacteriaceae studied. Serratia marcescens required 8 micrograms/ml to inhibit 90% of strains. Among gram-positive cocci, 50% of strains were inhibited by 2 micrograms/ml of cefodizime (including methicillin resistant Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus faecalis, and penicillin-resistant Streptococcus pneumoniae). Pseudomonas aeruginosa was less susceptible to cefodizime. Cefotaxime, an antibiotic closely related to cefodizime structurally, was about fourfold more active. PMID- 6295263 TI - Effects of novobiocin, coumermycin A1, clorobiocin, and their analogs on Escherichia coli DNA gyrase and bacterial growth. AB - Novobiocin, coumermycin A1, and clorobiocin, structurally related compounds that antagonize the B subunit of the essential bacterial enzyme DNA gyrase, were compared with 18 of their analogs for the inhibition of Escherichia coli DNA gyrase supertwisting activity in vitro and of bacterial multiplication. This family of compounds has a 4-hydroxy-8-methylcoumarin core substituted in the 7 and 3 positions. Important for enzyme inhibition in vitro is a 7 ether linkage to a 3'-substituted noviose sugar. The 3'-ester-linked 5-methylpyrrole, found in the coumermycin series, conferred at least 10-fold more inhibitory activity than did the similarly linked amide, found in the novobiocin series; lack of the pyrrole and amide results in the loss of inhibitory activity. Of many aryl and alkyl substituents linked as an amide at the 3 position, the 4-hydroxyl-3-(3-methyl-2 butenyl)benzoic acid moiety, found in novobiocin and clorobiocin, and the reduplication of the coumarin-noviose-5-methylpyrrole, found in coumermycin A1, were most effective in gyrase inhibition. In vivo, the ability of these compounds to inhibit the growth of E. coli varied greatly. The enhanced inhibition of gyrase in vitro conferred by a 5-methylpyrrole relative to an amide in the 3' noviose position was reflected in inhibition of bacterial multiplication. Several substitutions at the 3 position of the coumarin core conferring similar antagonism of gyrase in vitro resulted in substantially different inhibitory activities for E. coli, suggesting that these moieties at the 3 position affect drug access to the intracellular target. This target was shown for isobutyryl PNC NH2 (PNC-NH2 is 3-amino-4-hydroxy-8-methyl-7-[3-O-(5-methyl-2 pyrrolylcarbonyl)noviosyloxy] coumarin) and confirmed for novobiocin, coumermycin A1, and clorobiocin to be in the B subunit of DNA gyrase. PMID- 6295265 TI - Mechanism of action of dnacin B1, a new benzoquinoid antibiotic with antitumor properties. AB - Dnacin B1 preferentially inhibited the incorporation of [3H]thymidine into acid insoluble fractions in Escherichia coli. At a sublethal concentration, dnacin B1 caused filamentous growth in E. coli and induced prophage lambda. The antibiotic also showed potent bactericidal activity against repair-deficient E. coli strains, such as recA, recB, and polA strains, In in vitro studies, dnacin Ba raised the melting temperatures of various double-stranded DNAs. In addition, the antibiotic showed DNA-cleaving activity against PM2 DNA in the presence of reducing agents, and the activity was suppressed by scavengers for oxygen free radicals and an iron-specific chelator, desferrioxamine E. The stimulation of the generation of superoxide radical by dnacin B1 was confirmed by measuring the reduction of neotetrazolium. Therefore, it can be presumed that the primary cellular target of dnacin B1 is DNA in susceptible cells, and the autooxidation of DNA-bound dnacin B1 causes the generation of oxygen-free radicals that result in the damage of DNA and the inhibition of its synthesis. PMID- 6295266 TI - Interactions among amphotericin B, 5-fluorocytosine, ketoconazole, and miconazole against pathogenic fungi in vitro. AB - Interactions among amphotericin B, 5-fluorocytosine, ketoconazole, and micoconazole were tested for all possible paired and triple drug combinations and all four drugs combined against three isolates of Candida albicans, three Candida spp., two isolates of Cryptococcus neoformans, and three isolates Aspergillus fumigatus. An assay for inhibitory activity was developed in which growth in the presence of an antifungal agent was expressed as a percentage of the growth in drug-free cultures. For nearly all of the antifungal combinations, the interaction was additive against most fungal isolates. Drug combinations that included amphotericin B and ketoconazole were most often synergistic, i.e., amphotericin plus ketoconazole, amphotericin plus 5-fluorocytosine plus ketoconazole, and amphotericin plus 5-fluorocytosine plus ketoconazole plus miconazole, whereas the combination of ketoconazole plus miconazole showed the strongest tendency towards antagonism. The data in this screening survey provide a basis for further studies of drug interactions in vivo and in vitro. PMID- 6295267 TI - Diffusion of beta-lactam antibiotics through liposome membranes containing purified porins. AB - A method to determine the diffusion of cephalosporins through porin pores in vitro was developed, using liposomes reconstituted from phospholipids, lipopolysaccharides, and purified porin trimers. With this method, the roles of several species of porin pores from Escherichia coli and Salmonella typhimurium in the diffusion of cephalexin, cephaloridine, and cephalothin were examined. Results clearly showed that porins from E. coli B and 39,000-molecular-weight porins from S. typhimurium formed the most efficient pores. Thus, these were considered to represent a single functional group. OmpF and OmpE porins of E. coli K-12 and 38,000-molecular-weight porins of S. typhimurium formed moderately efficient pores. OmpC porins of E. coli K-12 and 40,000-molecular-weight porins of S. typhimurium were the least efficient pore formers. The present method can be used to distinguish the role of individual porin pores in the diffusion of cephalosporins. PMID- 6295268 TI - Pharmacokinetic characteristics of intravenous ceftriaxone in normal adults. AB - The multiple-dose pharmacokinetics and tolerance of intravenous ceftriaxone were investigated in 44 adults with normal renal function. Doses of 0.5, 1.0, and 2.0 g every 12 h and 2 g every 24 h were administered intravenously at a constant rate over 30 min. Plasma and urine samples were collected after the first (day 1) and last (day 4) dose and assayed for ceftriaxone by high-pressure liquid chromatography. Considering all four doses, mean peak plasma concentrations ranged from 79 to 255 micrograms/ml on day 1 and from 101 to 280 micrograms/ml on day 4. Trough concentrations at 12 h on day 1 were 15 to 45 micrograms/ml and 20 to 59 micrograms/ml on day 4. After a dose regimen of 2 g every 24 h, trough levels were still in the clinically therapeutic range (13 to 15 microgram/ml). The mean beta-phase t1/2 was markedly long (6.3 to 6.9 h) and was independent of dose. The fraction of dose excreted unchanged in the urine (0.33 to 0.44) indicated a substantial nonrenal mechanism of elimination. The plasma clearance ranged between 1,002 and 1,449 ml/h, and renal clearance ranged from 353 to 529 ml/h. The apparent volume of distribution varied from 9.2 to 13.5 liters. The dose-related increases in calculated Vd and Clp could be attributed to concentration-dependent plasma protein binding because of a larger free fraction of drug at higher concentrations. The drug was well tolerated, and no significant clinical or laboratory abnormalities were noted. PMID- 6295269 TI - Role of serum protein binding and multiple antibiotic doses in the extravascular distribution of ceftizoxime and cefotaxime. AB - The extravascular penetration of ceftizoxime and cefotaxime was studied in a rabbit subcutaneous Visking chamber model. Four rabbits, implanted with four chambers each, received each drug intramuscularly at a dose of 50 mg/kg every 3 hours for eight doses. Serum drug concentrations were measured after the eighth dose, and extravascular (chamber) concentrations were measured after the first and eighth doses. Cefotaxime (93% bound to rabbit serum proteins) demonstrated a much lower peak chamber-to-peak serum percent penetration after the first dose (20/163 = 13%) than did the less-bound (32%) ceftizoxime (21/52 = 40%, P less than 0.002). Similarly, the ratio of the chamber fluid area under the curve to the serum area under the curve was significantly lower for cefotaxime (15%) than for ceftizoxime (44%, P less than 0.002) after the first dose. Both agents approached equilibrium conditions between the intravascular and extravascular space by the eighth dose, and the ratios of chamber area under the curve to serum area under the curve of cefotaxime (76%) and ceftizoxime (79%) were similar. The peak-to-peak percent penetration of ceftizoxime (54%) was still significantly higher than that of cefotaxime (41%, P less than 0.01), although the chamber concentration of cefotaxime (66.2 micrograms/ml) was considerably higher than that of ceftizoxime (28.2 micrograms/ml). This study illustrates (i) dampened peak-to-trough antibiotic level fluctuation seen at extravascular sites as compared with measured serum concentrations, (ii) the large differences in extravascular penetration between single- and multiple-dose studies, and (iii) the importance of serum protein binding in the delay, but not the prevention, of extravascular drug distribution. PMID- 6295270 TI - Pharmacokinetics, protein binding, and extravascular distribution of ceftizoxime in normal subjects. AB - Thirteen normal subjects were given 2, 3, or 4 g of ceftizoxime intravenously in a prospective single-blinded study. Serum and urine concentrations were measured for 48 h. The beta-phase serum half-life ranged from 1.4 to 1.7 h. Approximately 100% of the agent was recovered unchanged in the urine over 48 h. Six of 13 subjects reported subjective complaints consisting of mild dizziness and moderate headache, all of which were transient. One subject also had a minimally elevated serum glutamic oxalacetic transaminase level 1 day after drug administration. Based on serum pharmacokinetics and serum protein binding determined in this investigation, unbound ceftizoxime appeared to distribute with the total body water. PMID- 6295271 TI - Activity of ketoconazole and its deacyl derivative against Plasmodium falciparum and Candida isolates. AB - Deacylketoconazole was 15- to 50-fold more active against Plasmodium falciparum than was ketoconazole, based on [(3)H]hypoxanthine uptake and quantitative parasite counts. In contrast, there were no significant differences between these drugs in their activity against clinical isolates of Candida spp. PMID- 6295272 TI - Isolation of bromovinyldeoxyuridine-resistant strains of herpes simplex virus and successful chemotherapy of mice infected with one such strain by using acyclovir. AB - Several strains of herpes simplex virus which were resistant to bromovinyldeoxyuridine were isolated by passaging the virus in the presence of the drug in tissue culture. The resistance of the majority of isolates was accounted for by their reduced ability to induce the enzyme thymidine kinase. These strains were co-resistant to acyclovir, but showed reduced pathogenicity in mice. However, another type of bromovinyldeoxyuridine-resistant virus was isolated which induced normal levels of thymidine kinase and retained virulence for mice. This resistant virus was sensitive to acyclovir and was successfully treated using oral acyclovir therapy. PMID- 6295273 TI - Efficacy of (E)-5-(2-bromovinyl)-2'-deoxyuridine against different herpes simplex virus strains in cell culture and against experimental herpes encephalitis in mice. AB - (E)-5-(2-Bromovinyl-2'-deoxyuridine (BrVUdR) showed strong antiviral activity against different laboratory strains and clinical isolates of herpes simplex virus type 1 (HSV-1) on primary rabbit testes (PRT) cells with a 50% inhibition of plaque formation (ID50) at 0.01-0.02 microM. One laboratory strain (HSV-1-S), however, was completely refractory even at concentrations as high as 100 microM. In contrast, the ID50S for all herpes simplex virus type 2 (HSV-2) strains were about 10(2) - 10(3) times higher (8-25 microM) than for the HSV-1 strains. No toxicity in mice treated with 140 mg BrVUdR/kg/day for 14 days was observed, and successful treatments of herpes encephalitis in mice induced experimentally by intracerebral infection with one laboratory strain (HSV-1-Kupka) and one clinical isolate (HSV-1-64) were achieved. Treatment of encephalitis in mice induced by the strain HSV-1-S insensitive to BrVUdR in cell culture failed to be effective. Similar antibody titers against HSV-1 were found in surviving mice of the control and of the BrVUdR-treated groups. PMID- 6295274 TI - Effects of nucleoside analogues on the expression of herpes simplex type 1 induced proteins. AB - Exposure of herpes simplex virus type 1 (HSV-1)-infected Vero cells to the nucleoside analogues 5-iodo-5'-amino-2',5'-dideoxyuridine (AIdUrd), 5-iodo-2' deoxyuridine (IdUrd) or 5'-amino-2',5'-dideoxythymidine (5'-AdThd) resulted in altered expression of HSV-1-induced proteins. Infected cell proteins (ICPs) synthesized in the presence of the nucleoside analogues were compared by sodium dodecyl sulphate (SDS) polyacrylamide gel electrophoresis to ICPs from non-drug treated cells and it was found that there was no effect on HSV-1-induced alpha proteins but beta and gamma proteins were reduced as much as 60%. There were three exceptions: ICP 35 (Mr = 46,000) and ICP 39 (Mr = 36,000) were not reduced and ICP 36 (Mr = 42,000) was increased during drug treatment. Progeny virions were isolated from drug-treated infected Vero cells and were compared to progeny isolated from control cells with respect to their polypeptide make-up and for their ability to induce HSV-1 proteins in non-drug-treated Vero cells. The progeny virus from drug-treated cells exhibited altered protein patterns on SDS polyacrylamide gels with respect to control HSV-1. The progeny virions from AIdUrd- or IdUrd- but not from 5'-AdThd-treated cells were defective in their abilities to induce proteins upon subsequent infection of non-drug-treated Vero cells. Two unusual phosphoproteins were detected; one with an apparent molecular weight of 30,000 was induced by progeny virus from AIdUrd-treated cells and another at approximately 69,000 was induced by progeny virus from 5'-AdThd treated cells. PMID- 6295275 TI - Isolation of indigenous enteroviruses from chemically treated and dewatered sludge samples. AB - Samples of wastewater sludge were examined for infectious enteroviruses before and after they had been chemically conditioned and dewatered. The least virus was recovered from the cake produced by filter pressing of sludge, which had a greatly increased solids content (39 to 45% [wt/vol]) relative to the untreated sludge (4.2 to 6.2% [wt/vol]) and in one plant was at pH 11 due to the lime conditioner used. Conditioning with a cationic polyelectrolyte before dewatering by centrifugation produced a watery sludge (2.7 to 5.3% [wt/vol]) from which high titers of infectious virus were recovered which were often greater than those isolated from the untreated sludge (0.6 to 1.4% [wt/vol]). This was thought to be due to saturation of virus and sludge floc adsorption sites by the polyelectrolyte, resulting in the liberation of virions from the sludge solids. PMID- 6295276 TI - Effects of temperature and salinity on Vibrio cholerae growth. AB - Laboratory microecosystems (microcosms) prepared with a chemically defined sea salt solution were used to study effects of selected environmental parameters on growth and activity of Vibrio cholerae. Growth responses under simulated estuarine conditions of 10 strains of V. cholerae, including clinical and environmental isolates as well as serovars O1 and non-O1, were compared, and all strains yielded populations of approximately the same final size. Effects of salinity and temperature on extended survival of V. cholerae demonstrated that, at an estuarine salinity (25%) and a temperature of 10 degrees C, V. cholerae survived (i.e., was culturable) for less than 4 days. Salinity was also found to influence activity, as measured by uptake of 14C-amino acids. Studies on the effect of selected ions on growth and activity of V. cholerae demonstrated that Na+ was required for growth. The results of this study further support the status of V. cholerae as an estuarine bacterium. PMID- 6295277 TI - Mechanisms of inactivation of poliovirus by chlorine dioxide and iodine. AB - Chlorine dioxide and iodine inactivated poliovirus more efficiently at pH 10.0 than at pH 6.0. Sedimentation analyses of viruses inactivated by chlorine dioxide and iodine at pH 10.9 showed that viral RNA separated from the capsids, resulting in the conversion of virions from 156S structures to 80S particles. The RNAs release from both chlorine dioxide- and iodine-inactivated viruses cosedimented with intact 35S viral RNA. Both chlorine dioxide and iodine reacted with the capsid proteins of poliovirus and changed the pI from pH 7.0 to pH 5.8. However, the mechanisms of inactivation of poliovirus by chlorine dioxide and iodine were found to differ. Iodine inactivated viruses by impairing their ability to adsorb to HeLa cells, whereas chlorine dioxide-inactivated viruses showed a reduced incorporation of [14C]uridine into new viral RNA. We concluded, then, that chlorine dioxide inactivated poliovirus by reacting with the viral RNA and impairing the ability of the viral genome to act as a template for RNA synthesis. PMID- 6295278 TI - Extraction of Clostridium perfringens spores from bottom sediment samples. AB - Two extraction-separation procedures were developed and evaluated for use in conjunction with the mCP membrane filter method for the enumeration of Clostridium perfringens spores in bottom sediments. In the more facile of the two procedures, a distilled-water suspension of the sediment sample is pulse sonicated for 10 s and allowed to settle. Portions of the supernatant are then removed for membrane filtration. This procedure is recommended for general use. The more complicated procedure is recommended for situations in which the presence of high levels of toxic materials is suspected or in which relatively low spore densities are present in fine silts. In this procedure, sonication is followed by a distilled water wash. The centrifuged sediment is resuspended in distilled water and mixed with the components of a two-phase separation system (50% polyethylene glycol in distilled water and 25% sucrose in 3 M phosphate buffer [pH 7.1]). After equilibration of the system and low-speed centrifugation, the top phase and interphase are removed, mixed, and membrane filtered. The recoveries of C. perfringens spores by the two procedures, when used in conjunction with the mCP method, were comparable to each other and significantly greater than those by the British most-probable-number method. It was estimated that more than 85% of the spores were recovered by the procedures. The precision of the sonicate-and-settle-mCP procedure was markedly better than that obtained theoretically by the most-probable-number method and approached that theoretically attributable to counting an average of 85 colonies on each of two plates. PMID- 6295279 TI - Purification and properties of two ribonuclease H enzymes from yeast. PMID- 6295280 TI - Carbohydrate heterogeneity of vesicular stomatitis virus G glycoprotein allows localization of the defect in a glycosylation mutant of CHO cells. PMID- 6295281 TI - Stimulation of liver glycogen particle synthase D phosphatase activity by caffeine, AMP, and glucose 6-phosphate. PMID- 6295282 TI - Functional subunit structure of photosystem 1 reaction center in Synechococcus sp. PMID- 6295283 TI - Dephosphorylation of skeletal muscle phosphorylase, glycogen synthase, and phosphorylase kinase beta-subunit by a Mn2+-activated protein phosphatase. PMID- 6295285 TI - A case of possible Penicillium tinea capitis. PMID- 6295284 TI - Polyphosphoinositide synthesis in rabbit erythrocyte membranes. PMID- 6295286 TI - Effects of dietary fibre and exercise on mid-morning diabetic control--a controlled trial. AB - Twenty-one insulin-dependent diabetic children completed a trial of 4 different breakfasts, given in random order. Three diets differed in fibre content. The fourth diet contained soya beans as part (38%) of the dietary fibre source. Children collected capillary blood samples on to filter paper strips which were analysed for blood glucose content. Each morning the children were asked to exercise vigorously for an hour and to rest for an hour, resulting in comparable rest and exercise periods for each child. Mean initial blood glucose levels on the 4 diets were not significantly different. The low-fibre diet resulted in the highest blood glucose concentrations after breakfast. Blood glucose levels on the high-fibre diet did not differ from those on the medium-fibre diet. The bean diet produced the lowest mean blood glucose level and the smallest reduction in blood glucose level in the hour before lunch. All the children found the bean diet unacceptable but liked the high- and medium-fibre diets, which were as popular as the low-fibre diet. The level of prescribed exercise had no effect on the level of blood glucose. It appears that the potentially major benefits from beans are limited by their unpalatability. The more acceptable cereal fibre produces a smaller but important benefit on morning hyperglycaemia after breakfast. PMID- 6295287 TI - Systemic candidiasis. PMID- 6295288 TI - Cytotoxic and hemolytic action of coal-quartz mixtures. AB - The effect of coal-quartz mixtures that contained graded amounts of coal or quartz was studied using peritoneal macrophage cultures and plasma-free washed sheep erythrocyte suspensions. A stoichiometric relationship was observed between the quartz content of coal-quartz mixture and the amount of lactic dehydrogenase liberated extracellularly and acid dye uptake by macrophage monolayers. Total lactic dehydrogenase of the monolayers plus supernatant remained unaltered following exposure to different dust mixtures. Dusts containing a higher concentration of quartz also produced a greater degree of hemolysis. Coal did not inhibit the quartz-induced macrophage cytotoxicity or erythrocyte hemolysis. PMID- 6295290 TI - Enzymatic synthesis and isolation of D-glucosamine-6-phosphate barium salt. PMID- 6295289 TI - Norwalk gastroenteritis associated with a water system in a rural Georgia community. AB - An outbreak of acute gastroenteritis occurred during January 4-9, 1982, in a rural community in north Georgia. A systematic telephone survey revealed that 63% of persons living in homes served by the community water system had symptoms of acute gastroenteritis in contrast to 9% of persons in homes served by private wells or other sources (P less than .001). A fourfold rise in antibody titer to the Norwalk virus occurred in 20 of 22 serum pairs obtained from ill persons. Fecal coliforms (greater than 16 MPN/100 ml) were detected in a spring which served as one water source for the community system. Surface runoff from a heavy rainfall, which preceded the outbreak, may have contaminated the system. Outbreaks of gastroenteritis should be promptly reported and investigated to facilitate corrective measures and to identify causative agents such as Norwalk virus. PMID- 6295291 TI - [Effect of psychotropic drugs administered during pregnancy on histochemical reactions in the fetal liver]. PMID- 6295292 TI - Particulate and solubilized LH receptor in the pig and bull testis. AB - Particulate and Triton X-100 solubilized receptors for luteinizing hormone (LH) in the pig and bull testis were studied. In some of the experiments tissue from the rat testis was included for comparison. LH receptors in the bull and pig revealed very similar association and dissociation kinetics, affinity (Kd = 1.2 1.8 x 10(-10) M), stability and physicochemical properties. Binding capacity was higher in the pig (73 fmol/mg protein) than in the rat (33 fmol/mg protein) and bull (17 fmol/mg protein). Hormone binding stabilized the receptor against thermal denaturation and temperature stability decreased upon solubilization. It is concluded that the properties of the testicular LH receptor of the pig and bull are very similar to those previously described from the rat. PMID- 6295293 TI - [The rabies endemic in the Ivory Coast as seen through the work of the Pasteur Institute of the Ivory Coast]. PMID- 6295294 TI - [Aspects of molecular biology of the rabies virus]. PMID- 6295295 TI - Mg++ antagonism of a prejunctional opiate receptor mediated effect at the frog neuromuscular junction. AB - The effect of meperidine on the spontaneous release of acetylcholine (mepp amplitude and frequency) and on the release evoked by nerve stimulation (epp amplitude and quantal content) was investigated in frog sciatic nerve-sartorius muscle in vitro. Meperidine (8.5 X 10(-5) or 1.6 X 10(-4)M) had no effect on mepp frequency recorded intracellularly in the absence of neuromuscular blockers, but reduced mepp amplitude. This depression was not antagonized by 3 X 10(-8) M naloxone and was explained by a nonspecific depressant action of meperidine on the endplate. When mepps and epps were recorded intracellularly in the presence of high Mg++, the mean amplitudes of both potentials were reduced equally by meperidine (either 1.6 or 4.2 X 10(-4)M) i.e. there was no change in the quantal content, and 3 X 10(-8)M naloxone failed to antagonize these amplitude depressions. When epps were recorded in the presence of d-tubocurarine, the percentage depression of epp amplitude by meperidine was significantly greater than found in the presence of Mg++ and was partially antagonized by naloxone indicating an opiate receptor mediated effect of meperidine. Thus it was concluded that Mg++ prevents this prejunctional opiate receptor mediated depressant effect of meperidine on the evoked release of acetylcholine. PMID- 6295296 TI - Inhibition by nifedipine of contraction and calcium entry in rat mesenteric arteries. PMID- 6295298 TI - Cross-reaction of antiserum against enterobacterial common antigen with Entamoeba histolytica HK-9 extracts. PMID- 6295297 TI - Nodular adrenal hyperplasia with elevated adrenocorticotropic hormone levels. AB - Micronodular adrenal hyperplasia is an uncommon adrenal disorder characterized by failure of urinary corticosteroid excretion to be suppressed by high-dose dexamethasone therapy. Thus, micronodular adrenal hyperplasia demonstrates dexamethasone suppressibility that resembles primary adrenal neoplasia. However, since some cases have been reported to have measurable plasma adrenocorticotropic hormone (ACTH) levels, it is unclear whether this disorder arises primarily in the pituitary-hypothalamic region or in the adrenal gland. Our patient had clinical features of Cushing's syndrome and elevated urinary corticosteroid excretion that did not suppress with even high doses of dexamethasone; however, ACTH levels were elevated and were suppressible with high-dose dexamethasone therapy. At operation, enlarged adrenal glands with multiple micronodules were found. This case is compatible with the hypothesis that hypothalamic-pituitary hyperfunction precedes the development of micronodular adrenal disease in some cases. PMID- 6295299 TI - [Ninth Seminar on Amebiasis. 9-11 November 1981]. PMID- 6295300 TI - The effects of prostaglandins on the secretion resulting from Entamoeba histolytica "enterotoxin". PMID- 6295301 TI - Adherence and ingestion of bacteria by trophozoites of Entamoeba histolytica. PMID- 6295302 TI - Separation of adherence, cytolytic, and phagocytic events in the cytopathogenic mechanisms of Entamoeba histolytica. PMID- 6295303 TI - [Isolation of clones of E. histolytica deficient in adhesion to human erythrocytes]. PMID- 6295304 TI - [Clones of E. histolytica deficient in phagocytosis present a deficiency in virulence]. PMID- 6295305 TI - [Morphology of trophozoites of Entamoeba histolytica on natural substrates. Study using scanning electron microscopy]. PMID- 6295306 TI - [Interactions between lectin and receptor mediate the adhesion of E. histolytica to epithelial cells. Relation of adhesion to the virulence of the strains]. PMID- 6295307 TI - [Detection of early cellular damage produced by Entamoeba histolytica in epithelia]. PMID- 6295308 TI - [Receptors participating in the adhesion of E. histolytica to human erythrocytes]. PMID- 6295309 TI - Virulence of Entamoeba histolytica upon continuous axenic cultivation. PMID- 6295311 TI - Phagosomal membranes from Entamoeba histolytica: isolation and biochemical properties. PMID- 6295310 TI - [Presence of collagenolytic activity in trophozoites of Entamoeba histolytica]. PMID- 6295312 TI - [Cytotoxic activity of Entamoeba histolytica on lymphoid cells of mouse spleen]. PMID- 6295313 TI - High salt SDS-DEP technic for isolation of "intact" rRNA from Entamoeba histolytica. PMID- 6295314 TI - [Interaction between Entamoeba histolytica and the cecal epithelium of the guinea pig. Quantitative study]. PMID- 6295315 TI - Activation of lymphocytes from healthy donors and patients with amebiasis by extracts of Entamoeba histolytica. PMID- 6295316 TI - Antigen fraction from Entamoeba histolytica strain HK9 for use in ELISA. PMID- 6295317 TI - [Excretion of anti-ameba specific antibodies of class IgA in bile from rats immunized with trophozoites of Entamoeba histolytica cultivated in axenic media]. PMID- 6295318 TI - The nitroblue tetrazolium (NBT) reduction of Entamoeba histolytica during endocytosis of E. coli and homologous antibodies. PMID- 6295319 TI - [Effect of cycloheximide on the inhibition of chemotaxis in human monocytes caused by products of E. histolytica]. PMID- 6295320 TI - [Partial purification of several surface antigens of E. histolytica. Investigation of the biological activity with respect to sera from patients with amebic liver abscess]. PMID- 6295321 TI - RNA depolymerase in Entamoeba histolytica: soluble vs ribosomal. PMID- 6295323 TI - [Identification of circulating amebic antigen in man by immunoenzymatic analysis. I. Development of the technic]. PMID- 6295322 TI - [Monoclonal antibodies against Entamoeba histolytica]. PMID- 6295324 TI - [Detection of amebic antigen by the ELISA method in sera of patients with hepatic abscess]. PMID- 6295325 TI - [Long term detection of anti-amebic antibodies by counterimmunoelectrophoresis and indirect hemagglutination]. PMID- 6295326 TI - [Localization of actin in trophozoites of Entamoeba histolytica (HMI)]. PMID- 6295327 TI - In vitro activity of certain quassinoid anti-tumor agents against Entamoeba histolytica. PMID- 6295328 TI - [Immunochemistry of a lipopeptidophosphoglycan extracted from trophozoites of Entamoeba histolytica strain HK-9 cultivated in axenic media, using the phenol water method]. PMID- 6295329 TI - [Localization of a lipopeptidophosphoglycan extracted by phenol-water from trophozoites of the HK-9 strain of Entamoeba histolytica]. PMID- 6295330 TI - Specificity of iron requirements of Entamoeba histolytica in vitro. PMID- 6295331 TI - Protein biosynthesis by Entamoeba histolytica in culture. PMID- 6295332 TI - [Clinical correlation of zymodemes of E. histolytica]. PMID- 6295334 TI - A review of isoenzyme characterization of Entamoeba histolytica with particular reference to pathogenic and non-pathogenic stocks isolated in Mexico. PMID- 6295333 TI - Observations on zymodeme studies of Entamoeba histolytica in Durban, South Africa. PMID- 6295335 TI - Membrane dynamics of Entamoeba histolytica and Giardia lamblia studied with reflection contrast microscopy and the carbon immunoassay. PMID- 6295336 TI - [Interaction between zinc and dietary factors]. PMID- 6295337 TI - Spinal leptomeningeal invasion from intracranial glioblastoma multiforme. AB - The cases of 2 patients with meningeal and spinal spread of primary intracranial glioblastoma are reported. While such spinal meningeal spread of intracranial glioblastoma is considered rare, it appears that such involvement is not uncommon if specifically investigated. The incidence of meningeal seeding by primary intracranial glioblastoma may be expected to rise as new therapies are directed toward the primary tumor. PMID- 6295338 TI - [Granular cell tumors of the oral cavity]. PMID- 6295339 TI - Third-generation cephalosporins for polymicrobial surgical sepsis. AB - During 31 months of study, 808 patients with polymicrobial surgical infection were randomized for antibiotic therapy between a third-generation cephalosporin (moxalactam disodium [149], cefotaxime sodium [125], and cefoperazone sodium [141]) and the combination of gentamicin sulfate plus clindamycin (393). Results based on antibiotic therapy included the following: cure in 83% given cephalosporin, 73% with antibiotic combination; control but recurrent sepsis in 7% and 15%; and failure in 4% and 8%, respectively. Such data support the tenet that third-generation cephalosporins are at least equal, if not superior, to the combination of gentamicin plus clindamycin for treatment of polymicrobial surgical sepsis. PMID- 6295340 TI - Ranitidine reverses cimetidine-induced mental confusion in a patient with Zollinger-Ellison syndrome. PMID- 6295341 TI - [Inhibitory effect of noradrenaline on cholinergic neurotransmission in an isolated guinea-pig tracheal muscle]. PMID- 6295342 TI - [Comparison of bronchodilating and metabolic effects of intravenous aminophylline and subcutaneous epinephrine individually and in combination in patients with bronchial asthma]. PMID- 6295343 TI - [On the induction of asthma in guinea pigs by toluene diisocyanate (TDI)]. PMID- 6295344 TI - [Ultrastructural characteristics of the differentiation of glandular cells in the anterior pituitary gland of collared lemmings (Dicrostonyx) from the Vrangel Island]. AB - In 15.5-, 16-, 17.5-, 18.5-, 20-day-old fetuses and newborn lemmings, the ultrastructure of the anterior lobe of the hypophysis has been studied. In the parenchyma of the 15.5-day-old fetus gland, some cells containing secretory material are revealed, but they are difficult for identification, since differentiation of their organellas is not completed and polymorphism of their granules is strongly manifested. On the 17.5th day of the prenatal development TTH-, ACTH-, LH-, STH-secreting adenocytes are detected; they include secretory granules having diameters 70--100 nm, 80--150 nm, 60--140 nm and 200--400 nm, respectively. On the 18.5th day FSH-secreting cells are identified. Lactotropocytes appear on the 20th day. The anterior lobe of the hypophysis of 17.5--18.5-day-old fetuses are characterized by a high level of the secretory activity which decreases to some extent before birth. Ultrastructure of the cellular organellas in the newborn hypophysis is similar to that in a mature animal, nevertheless, the diameter of the secretory granules does not reach the size of these elements in mature individuals. PMID- 6295346 TI - [Complex light and electron microscope study of connections in the central nervous system. Review of current methodological approaches]. PMID- 6295345 TI - [Relations between pigmentation of the cornea and the number of epidermal melanophores in Rana temporaria L. larvae]. AB - The dynamics of the external cornea pigmentation in Rana temporaria L. larvae at the 22d developmental stage have been studied under conditions favourable for various course of certain morphological reactions in the pigment system. The cornea together with the surrounding skin is transferred on the dorsal surface of the larva body, and the piece of the dorsal surface skin is put instead of the cornea removed. When using the reciprocal transplantation method and preserving the organism's integrity (without disturbing melanocyte-stimulating source- namely, the hypophysis, and melatonine sources--namely, the pineal gland and the lateral eyes) the corneal pigmentation is observed on the background of perfect morphological reactions in the pigment system, while the larvae are maintained on the dark and light substrates, that is at various density of the pigment cells (120 larvae have been used). The pigmentation dynamics have been studied from the 6th up to the 20th day in total preparations. The epidermal melanophores density is estimated in 4 areas of each preparation. The melanin amount is estimated by means of the electron paramagnetic resonance-spectrometry according to the contents of free radicals expressed in relative units. A direct proportional dependence between the significantly higher melanin contents (1.5-fold) and a significantly quicker (1.5-fold) process of the corneal pigmentation is revealed, that agrees with an increasing number of the pigment cells per one unit of the body surface in the larvae maintained on the dark substrate. In the larvae maintained on the light substrate, the dependence is of a reverse character. It is probable that the factors forcing the pigmented cells, at cultivation the neural crest cells in vitro to reject from each other, affect the pigmentation of the larval cornea in vivo. If it is the case, the processes specific for the embryonal period, transgress during the cornea pigmentation at the larval stages of development. PMID- 6295347 TI - Evaluation of eight cases of early gastric cancer. AB - We review eight cases of early gastric cancer which occurred in five female and three male patients. In seven patients the main complaint was epigastric pain and in one melena. In seven of the eight cases, endoscopic diagnosis was established on the first examination and confirmed by biopsy. Types of early gastric cancer observed were: type I, one case; IIa, one case; IIc three cases: III, two cases and type III + IIc, one case. In five cases the neoplasia was located in the antrum, two in the pre-pyloric region, two along the greater curvature and one in the anterior wall. Three cases were located in the body of the stomach, two in the lower third of the lesser curvature and one in the upper third of the greater curvature. From the histopathologic standpoint four cases were differentiated adenocarcinomas, three undifferentiated adenocarcinomas and one case mucus carcinoma. In no case did we encounter regional lymph node metastases. Postoperative survival rates at the present time varies between one and five years. PMID- 6295348 TI - [Effect of the dietary fiber content in nutrition on various stool parameters, in man]. AB - Ten subjects were studied for two periods of six days each during which they were fed respectively a regular diet (control diet) followed by a fiber-supplemented diet (experimental diet). The experimental diet consisted of a regular diet in which some foods where substracted and some bran biscuits, specially produced for this investigation, were added. The experimental diet produced a significant increase in daily stool weight, dried stool weight, water contents of the stools and intestinal transit time. The study has shown that special bran biscuits is effective in promoting a increase in bowel transit time, and therefore should be submitted to therapeutic trial in patients with constipation. PMID- 6295349 TI - Pathologic quiz case 2. Adenocarcinoma of the nasal cavity. PMID- 6295350 TI - Autoregulation of the modified low density lipoprotein receptor in human monocyte derived macrophages. AB - Regulation of the macrophage receptor for modified low density lipoprotein (LDL) was evaluated using human monocyte-derived macrophages and acetyl LDL. Factors that regulate native LDL receptor activity in other cell types, such as the cholesterol content of the incubation medium, insulin, and platelet-derived growth factor had no effect on acetyl LDL degradation. Conditioned medium from mature macrophages significantly stimulated acetyl LDL degradation and enhanced cholesterol esterification by freshly isolated monocytes. Time course studies indicated that increasing time in culture was associated with increasing potency of the stimulating activity of macrophage-conditioned medium. These data suggest that a macrophage secretory product may be a prime modulator of modified LDL receptor activity on monocyte-macrophages, thus exerting an autocrine regulatory effect. The positive autoregulation of the human macrophage receptor for modified LDL could accelerate cellular cholesteryl ester accumulation and macrophage derived foam cell formation. PMID- 6295351 TI - Diatomite as a source of cristobalite for dental investments a differential thermal analysis study. AB - Cristobalite can be obtained from diatomite by heat-treatment at relatively low temperatures and short times without the use of flux. The differential thermal analysis method used to examine the alpha leads to beta inversion was not as satisfactory as it might have been, and to determine the amount and crystallinity of cristobalite formed any such method should be supported by other techniques, in particular, X-ray diffraction. To determine the suitability or otherwise of cristobalite produced from diatomite for use in dental investments, dialatometric studies are indicated since the property of thermal expansion is of fundamental importance to a dental investment. PMID- 6295352 TI - A probable linkage between familial Paget's disease and the HLA loci. PMID- 6295354 TI - Enzyme linked immunosorbent assay for detecting specific IgM antibody in congenital cytomegalovirus infection. PMID- 6295353 TI - Double-blind controlled trial of ranitidine versus cimetidine in the treatment of duodenal ulceration. PMID- 6295355 TI - Improved specificity of ELISA for rotavirus. PMID- 6295356 TI - Jaundice and other drug reactions to ketoconazole. PMID- 6295357 TI - Opioid and nonopioid forms of stress-induced analgesia: some environmental determinants and characteristics. PMID- 6295358 TI - Neuroendocrine modulation of memory during development. PMID- 6295359 TI - [Economic problems and contradictions in mineral fertilizer application for solving nutrition problems in the developing countries]. AB - The optimum application of mineral fertilizers in the agriculture of the developing countries is inhibited by the monopolization of the mineral fertilizer industry in the capitalist economic world system and the numerous contradictions in the developing countries themselves. Thus it requires the full state sovereignty and economic independence of the developing countries, and the creation of such social conditions which enable the further extension and better utilization of the fertilizer capacities as well as the optimum fertilizer application. This means applying mineral fertilizers in the best possible way in order to ensure the nutrition of their own population and aiming at agrarian structures which promote mineral fertilization, restructuring the system of mineral fertilizer prices, and developing the mineral fertilizer service. PMID- 6295360 TI - [Storage and preservation of antigen for the enzyme-linked immunosorbent assay (ELISA) in the detection of antibodies against chicken bronchitis virus]. PMID- 6295361 TI - Chemical deglycosylation of ovine pituitary lutropin. A study of the reaction conditions and effects on biochemical, biophysical and biological properties of the hormone. AB - The oligomeric glycoprotein hormone, ovine lutropin was treated with anhydrous HF at 0 degrees C for 30, 60 and 180 min and at 23 degrees C for 60 and 180 min. The products, designated deglycosylated lutropin 1 (DGLH-1) to deglycosylated lutropin 5 (DGLH-5) respectively, were characterized by gel filtration, concanavalin A-Sepharose binding, disc electrophoresis, amino acid analysis, carbohydrate composition and spectral properties. The preparations were also evaluated for receptor binding activity and immunological activity and bioassayed in vitro in collagenase-dispersed rat interstitial cells. In DGLH-1, fucose and galactosamine were removed completely, and there was a 94% decrease in hexoses and 39% decrease in N-acetylglucosamine. Reaction with HF at 0 degrees C for 1 or 3h led to removal of all hexoses and additional loss of hexosamines. Reactions at 23 degrees C for either 1 or 3h were not of additional value in deglycosylation and none of the reaction conditions yielded the apohormone. All the five deglycosylated hormone preparations were not retained on immobilized-concanavalin A columns and on Sephadex G-100 they were eluted with an increased V(e)/V(0) ratio consistent with the loss of carbohydrate residues. Loss of all but the last of the N-acetylglucosamine residues decreased the abnormality of lutropin on sodium dodecyl sulphate/polyacrylamide-gel electrophoresis, but did not eliminate it. Receptor binding activities of DGLH-1 and DGLH-2 were not different from that of the native hormone, but that of DGLH-3 was slightly decreased and the products obtained at 23 degrees C (DGLH-4 and DGLH-5) had lower activity. Immunoreactivities followed a similar pattern. None of the derivatives had activity in the bioassay in vitro. All of the five derivatives inhibited the action of the native hormone in the bioassay in vitro. Their hormonal antagonistic activity was consistent with the receptor binding activity, with DGLH-5 being the least potent in this respect. The DGLH-4 and DGLH-5 preparations had undergone conformational changes as revealed by 8-anilinonaphthalene-1 sulphonate fluorescence, but this did not result in loss of quaternary structure. PMID- 6295363 TI - The autoreducible cytochromes c of the methylotrophs Methylophilus methylotrophus and Pseudomonas AM1. AB - The two types of soluble cytochrome c (cytochrome cH and cytochrome cL) found in methylotrophs are completely distinct proteins; one type is not a dimer or degradation product of the other. Free thiol groups are probably not involved in the unusually rapid autoreduction of the cytochromes at high pH. The axial ligands to the haem iron, histidine and methionine, are the same as in other low spin cytochromes c. The methionine ligand is displaced at high pH by an alternative strong-field ligand. This displacement does not occur on reduction of cytochrome cL by methanol dehydrogenase, but this does not rule out the possibility that the autoreduction mechanism is involved in the interaction of the dehydrogenase and cytochrome c. PMID- 6295362 TI - Submitochondrial localization and asymmetric disposition of two peripheral cyclic nucleotide phosphodiesterases. AB - There are two distinct cyclic AMP phosphodiesterases associated with the liver mitochondrion: one with the outer membrane and one with the inner membrane. No activity is associated with the lysosomal fraction. Both of the enzymes are peripheral proteins and can be released from the membranes by high-ionic-strength treatment. Treatment of intact mitochondria with trypsin and insoluble trypsin localizes these enzymes to the cytosol-facing surface of their respective membranes. The enzymes differ in regard to sedimentation coefficient, thermostability and susceptibility to inactivation by trypsin. Both enzymes degrade cyclic AMP and cyclic GMP. Whereas the outer-membrane enzyme displays Michaelis kinetics and appears to be a low-affinity enzyme, the inner-membrane enzyme displays kinetics indicative of apparent negative co-operativity. PMID- 6295364 TI - The nature of haem a3 in the oxidized state of cytochrome c oxidase. Evidence from low-temperature magnetic-circular-dichroism spectroscopy in the near infrared region. AB - The magnetic-circular-dichroism (m.c.d.) spectra of oxidized 'resting' bovine cytochrome c oxidase and the cyanide-inhibited form are reported at 5.15 T and at 4.2 K along with m.c.d. magnetization curves plotted at selected wavelengths. In both spectra there are features at 790nm and 1564nm due to Cua and haem a respectively, the e.p.r.-detectable components of the enzyme. There is a new peak at 1946nm only in the spectrum of the cyanide-inhibited enzyme. Arguments are advanced that assign this to low-spin ferric haem a3 bridged to Cua3, thereby forming a ferromagnetically coupled pair of metal ions. PMID- 6295365 TI - Ecto-5'nucleotidase and adenosine deaminase activities of lymphoid cells. PMID- 6295366 TI - Generation of free radicals and initiation of radical reactions in nitrones - Fe2+ - phosphate buffer systems. PMID- 6295368 TI - Evidence for plastoquinol-cytochrome f/b-563 reductase as a common electron donor to P700 and cytochrome oxidase in cyanobacteria. PMID- 6295367 TI - Cloning of yeast glycolysis genes by complementation. PMID- 6295369 TI - Phosphorylation of high mobility group proteins 14 and 17 by nuclear protein kinase NII in rat O6 glioma cells. PMID- 6295370 TI - A cyclic AMP dependent protein kinase in Dictyostelium discoideum. PMID- 6295371 TI - Multireceptor-induced release of adrenocorticotropin from anterior pituitary tumor cells. PMID- 6295372 TI - 14N-ENDOR evidence for imidazole coordination in copper proteins. PMID- 6295373 TI - Isolation and properties of the cytochrome B-C1 complex from Rhodopseudomonas sphaeroides. PMID- 6295374 TI - A 1H and 13C N.M.R. study of all-trans retinal in dodecyldimethylamine oxide micelles. PMID- 6295375 TI - Evidence contradicting the notion that gonadal hormones regulate brain opiate receptors. PMID- 6295376 TI - Increased nucleoside triphosphatase activity of rat liver nuclear matrix following low dose carcinogen intoxication. PMID- 6295377 TI - Inhibition of adriamycin-stimulated microsomal lipid peroxidation by mitoxantrone and ametantrone, two new anthracenedione antineoplastic agents. PMID- 6295378 TI - Direct proton and natural abundance carbon-13 NMR observation of liver changes induced by ethionine. PMID- 6295380 TI - Identification of [3H] histamine binding sites on gastric mucosal cells unrelated to histamine H2-receptors. PMID- 6295379 TI - Ovine corticotropin-releasing factor stimulates the concomitant secretion of corticotropin, beta-lipotropin, beta-endorphin and gamma-melanotropin by the bovine adenohypophysis in vitro. PMID- 6295381 TI - Does glucocorticoid deprivation promote the expression of adenosine receptor sites stimulating adenylate cyclase in rat adipocyte membranes? PMID- 6295382 TI - Unambiguous identification of the nickel EPR signal in 61Ni-enriched Desulfovibrio gigas hydrogenase. PMID- 6295383 TI - Effect of substrate size on tonin activity. PMID- 6295384 TI - Expression of early viral genes: a possible pre-alpha protein in cells infected with herpes simplex virus. PMID- 6295385 TI - Leukotriene B4 induces human suppressor lymphocytes. PMID- 6295386 TI - Genetic organization and cloning of Kirsten murine sarcoma virus DNA. PMID- 6295387 TI - Characterization of dopamine receptors by 3H-ADTN binding in calf adrenal zona glomerulosa. PMID- 6295388 TI - The presence of 5'-nucleotidase in swiss chard chloroplasts. PMID- 6295389 TI - Activation by thymidine kinase and potent antiherpetic activity of 2'-nor-2' deoxyguanosine (2'NDG). PMID- 6295390 TI - Diversity of nuclear phorbol ester tumor promoter receptors in mouse liver: evidence for two classes of binding sites. PMID- 6295391 TI - 1 alpha,25-Dihydroxyvitamin D3 suppresses proliferation of murine granulocyte macrophage progenitor cells (CFU-C). PMID- 6295392 TI - Generation of hydroxyl radical and its involvement in lignin degradation by Phanerochaete chrysosporium. PMID- 6295393 TI - Fine structure of the recB and recC gene region of Escherichia coli. PMID- 6295394 TI - Phosphorylation of the Mr = 34,000 protein in normal and Rous sarcoma virus transformed rat fibroblasts. PMID- 6295396 TI - Genetic changes in yeast cells cotransformed with mitochondrial DNA and plasmid YEp13 are not elicited by recombinant molecules made by covalent insertion of mitochondrial DNA into Yep13. PMID- 6295395 TI - Evidence against direct transfer of the adenine nucleotides by the heart mitochondrial creatine kinase-adenine nucleotide translocase complex. PMID- 6295397 TI - Structural differences between insulin and somatomedin-C/insulin-like growth factor-1 receptors revealed by autoantibodies to the insulin receptor. PMID- 6295398 TI - Small angle neutron scattering of the mitochondrial ADP/ATP carrier protein in detergent. PMID- 6295399 TI - Effects of monensin on the processing of pro-opiomelanocortin in the intermediate lobe of the rat pituitary. PMID- 6295400 TI - Chemical modification of opiate receptors with ethoxyformic anhydride and photo oxidation: evidence for essential histidyl residues. PMID- 6295401 TI - Injection of proteins into primary rat hepatocytes by erythrocyte-mediated techniques. PMID- 6295403 TI - Polyglucose content in the cell and the rate of glucose consumption during synchronous growth of Escherichia coli. PMID- 6295402 TI - Metabolites of thyrotropin releasing hormone inhibit angiotensin converting enzyme in vitro. PMID- 6295404 TI - (+)-Cyanidanol-3 changes functional properties of collagen. AB - About 6-7 (+)-cyanidanol-3 molecules are bound per collagen alpha-chain. The (+) cyanidanol-3 treated collagen contains an increased number of pepsin-resistant cross-links, is less susceptible to attack by mammalian collagenase, has a higher shrinkage temp and forms unstructured aggregates. Cell and organ culture studies show that these biological systems produce less protein and collagen in the presence of (+)-cyanidanol-3 and that the newly synthesized collagen is less soluble. PMID- 6295405 TI - Role of cyclic AMP in the inhibition of human platelet aggregation by quercetin, a flavonoid that potentiates the effect of prostacyclin. AB - Quercetin (3.3',4',5,7-pentahydroxyflavone) has previously been shown to inhibit cyclic nucleotide phosphodiesterases prepared from various cell homogenates and the function of intact human platelets. We now report that (1) high concentrations of quercetin raise platelet cAMP levels; and (2) quercetin potentiates the inhibitory effect of prostacyclin (PGI2) on ADP-induced washed human platelet aggregation and the elevation of platelet cAMP levels elicited by PGI2. These results suggest a role for cAMP in the mechanism of action of quercetin on blood platelets. PMID- 6295406 TI - Drug-induced lipid peroxidation in mice--II. Protection against paracetamol induced liver necrosis by intravenous liposomally entrapped glutathione. AB - If injected intravenously 2 hr before the drug, a dose of more than 175 mg/kg body weight glutathione (0.57 mmol/kg) protected male mice from acute liver necrosis induced by intraperitoneal administration of 400 mg/kg (2.65 mmol/kg) paracetamol. Soluble glutathione yielded a limited, and liposomally entrapped glutathione an optimal dose-dependent protective effect against drug-induced lipid peroxidation (as measured by in vivo ethane exhalation) liver necrosis (assessed by serum transaminases) and hepatic glutathione depletion (determined post mortem). N-Acetylcysteine solution had no effect in this model. PMID- 6295407 TI - Generation of hydroxyl radical by anticancer quinone drugs, carbazilquinone, mitomycin C, aclacinomycin A and adriamycin, in the presence of NADPH-cytochrome P-450 reductase. AB - The generation of hydroxyl free radicals in the system consisting of purified NADPH-cytochrome P-450 reductase and anticancer quinone drugs, such as carbazilquinone, mitomycin C, aclacinomycin A and adriamycin, has been confirmed by two methods. In the spin trapping study, using N-tert-butyl-alpha phenylnitrone as the spin trapping agent, four drugs generated hydroxyl radical trapped signals, and the formation of the spin adduct was dependent on time and the enzyme concentration. Among the four drugs, the generation time of signal was in the order of carbazilquinone, aclacinomycin A, adriamycin and mitomycin C, but the magnitude of signal intensity was different. In both aclacinomycin A and adriamycin, the signal disappeared in a few minutes. Catalase completely inhibited the formation of the spin adduct, while superoxide dismutase did not significantly inhibit, but effected in some manner. The generation of hydroxyl radical was also confirmed by the ethylene production from methional. Among the four drugs, the order of the magnitude of ethylene production was different from that of signal intensity by ESR study. Catalase potently inhibited the ethylene production, while superoxide dismutase effected in some manner. From these results, the interactions of anticancer quinone drugs with NADPH-cytochrome P-450 reductase and oxygen, and the possible relations of the enzymes to the radical related actions of these drugs are discussed. PMID- 6295408 TI - Clonidine effect on hepatic cGMP levels in vivo could be mediated by alpha 1 adrenoceptors. PMID- 6295409 TI - The effect of various calmodulin inhibitors on the response of adrenal glomerulosa cells to angiotensin II and cyclic AMP. PMID- 6295410 TI - [Effect of memantine on membrane properties and postsynaptic potentials in neurons of Aplysia californica]. AB - The effect of 1,3-dimethyl-5-aminoadamantane (DMAA, D-145, memantine, Memantine) in the concentration range of 10(-6) to 10(-3) mol/l was investigated on membrane properties and synaptic transmission by the current or voltage clamp method on neurons in the visceral ganglion of Aplysia california. In the lower range (concentrations 10(-6) to 10(-5) mol/l) the substance increases the speed of voltage changes during the course of the action potential, increases spontaneous synaptic activity, and induces a slight membrane depolarization. With increasing concentration the effects reverse in that slope of action potential is slowed and synaptic transmission delayed or blocked. At the RC-R15 synapse, isolated from its interneuron, 10(-5) mol/l DMAA evokes spontaneously and regularly firing EPSPs; 10(-3) mol/l blocks both, spontaneous activity, and its responsiveness to electrical stimulation. PMID- 6295411 TI - Cefotaxime in human lung tissue. AB - In the course of thoracic surgery in 79 patients lung tissue samples were taken up to around 2 and 4 h, respectively, after i.v. injection of 1 (n = 22) or 2 g cefotaxime (n = 57). In these samples cefotaxime levels were measured using a microbiological method. About 1 h after injection 1.6 and 5.1 microgram/g (median values), respectively, were found. Even later after injection cefotaxime could still be detected. These results are evaluated with respect to the problem encountered in the biological measurement of concentrations of ester cephalosporins in tissue due to desacetylation by hemolytic samples. PMID- 6295412 TI - [Mammalian collagenases: physiological and pathophysiological aspects]. AB - Biochemical and cell biological aspects of synthesis, activation and action of mammalian collagenases are presented and discussed with respect to collagen resorption under normal and pathological conditions. Particularly emphasized are the regulatory mechanisms involved which operate at the level of synthesis of the proenzyme, during activation of the latent enzyme, and by the presence of inhibitors of the active collagenase as well as by substrate specificity for the various collagen types. PMID- 6295413 TI - Bacterial collagenases and their clinical applications. AB - Clostridium histolyticum cultures secrete a specific collagenase which can be precipitated from the medium in relatively large amounts. The enzyme is a metalloproteinase capable of cleaving native collagen types I, II, III, IV and V. In addition to being a valuable tool in the laboratory the bacterial collagenase has found clinical applications in the treatment of third degree burns and decubitus, diabetic or arterial ulcers. Several cases of successful topical use of the crude enzyme in an ointment base are illustrated. More recently direct injection of a highly purified form of the enzyme has been proposed in the treatment of herniated discs and keloids and as an adjunct in vitrectomy. Preliminary results of the two latter interventions are described. PMID- 6295414 TI - Intervertebral discolysis with collagenase. AB - Animal experimental studies are reported in which the enzyme collagenase was used to digest the nucleus pulposus of the intervertebral disc in both dogs and monkeys. Studies in the same animals indicated a low risk toxicity. Clinical studies were started in 1979, in which the results in 82 patients are reported. All patients were subjected to rigid standards of selection. Of 78 patients followed, 80.4% had good results, 4.9% fair results, and 14.7% had poor results. There were no instances of toxicity. A double-blind study of 30 patients indicates clearcut superiority of collagenase over placebo in relieving the symptoms of herniated disc. PMID- 6295415 TI - [Influence of (+)-Catechin and Dithiocarb on the Pharmacokinetics of Phenprocoumon, Tolbutamide and Three Inhalation Anesthetics in rats]. AB - Pretreatment with (+)-catechin ((+)-cyanidanol-3, Catergen) (200 mg/kg p.o.) did not alter the elimination of intravenously injected phenprocoumon (0.6 mg/kg) or tolbutamide (100 mg/kg) in rats, while dithiocarb (Sodium diethyl dithiocarbamate) (200 mg/kg p.o.) prolonged only the elimination half-life of phenprocoumon to a small extent. The metabolism of halothane (100 ppm), enflurane (100 ppm) or methoxyflurane (300 ppm) was studied by measuring the disappearance of the compounds from the atmosphere of a closed exposure system. Pretreatment with (+)-catechin did not impair the metabolic removal of all three anesthetics; in the case of enflurane (+)-catechin caused a small, but significant shortening of the elimination half-life, for enflurane and methoxyflurane the uptake of the compounds into the rats seemed to be impaired under the influence of (+) catechin. Dithiocarb strongly impaired the in vivo metabolism of halothane and methoxyflurane, whereas that of enflurane remained unaffected. PMID- 6295416 TI - [Study of the effect of a dexamethasone-containing antirheumatic on the hypothalomus-adenohypophysis-adrenocortical system]. AB - The aim of this study was to investigate the effect of an analgesic-antirheumatic fixed combination drug containing dexamethasone (Ambene) on this endocrine system. 10 healthy volunteers received an intramuscular injection of Ambene as either single dose once, single dose on three consecutive days or on three alternating days. Early morning ACTH plasma concentrations, endogenous cortisol profiles and dexamethasone kinetics were determined and insulin-hypoglycemia tests stimulated stress conditions before, during and after Ambene-application. In all cases treatment led to short-term suppression of the reactability of the Hypothalamus-Anteropituitary Corticoadrenal System. Three days after single dose or alternating day administration for three days reactions of this endocrine system were completely restored, while after administration on three consecutive days there still was a slight decrease in basal and insulin-stimulated cortisol values. The short half-life of water soluble dexamethasone in Ambene appears to be the reason for the rapid restitution of this endocrine system. PMID- 6295417 TI - Sonographic and radiographic images. PMID- 6295418 TI - [Diagnostic methods for myocardial infarct in the patient undergoing aorto coronary surgery]. PMID- 6295419 TI - [Follow-up with 99mTc-pyrophosphate and myoglobin in myocardial necrosis]. PMID- 6295420 TI - Platelet reactivity and its dependence on alpha-adrenergic receptor function in patients with ischaemic heart disease. AB - We studied 57 patients admitted to hospital with ischaemic heart disease, including nine patients with variant angina, to evaluate platelet reactivity and its dependence on alpha-adrenergic receptor function. The threshold concentration for biphasic platelet aggregation in response to adrenaline and adenosine diphosphate was measured in fresh platelet rich plasma. There were age related alterations in platelet responsiveness to adrenaline. In 27 age matched control subjects platelets showed adrenaline induced aggregation at a concentration higher than 0.1 mumol. The threshold concentrations for adrenaline and adenosine diphosphate were 0.91 mumol and 4.68 mumol. In 16 patients with acute infarction, 14 with old infarction, nine with effort angina, and nine with rest angina, mean values of platelet aggregation threshold for both adrenaline and adenosine diphosphate were not altered significantly when compared with control subjects. In contrast, the values for adrenaline and adenosine diphosphate in nine patients with variant angina were 0.012 mumol and 2.24 mumol and seven of them showed obvious platelet hyperactivity to adrenaline at a concentration lower than 0.1 mumol. The threshold concentration for adrenaline induced aggregation did not correlate with serum cholesterol and triglyceride levels. PMID- 6295421 TI - Effect of enflurane on cerebellar cGMP and on motor activity in the mouse. AB - The effect of enflurane on the cerebellar content of the intracellular mediator cyclic 3', 5'-guanosine monophosphate (cGMP) and on motor activity was studied in mice. Seizures, as an index of increased motor activity, associated with an increase in cerebellar cGMP content were induced with isoniazide or picrotoxin. Enflurane 0.28-1.68 vol% produced a dose-dependent, reversible decrease in cerebellar cGMP (by about 50% at 0.28 vol%) and delayed or prevented both the increase in cerebellar cGMP and the convulsions induced by isoniazide. Enflurane also protected against picrotoxin-induced convulsions, but not against strychnine induced convulsions which presumably do not involve cerebellar mechanisms. These results indicate that enflurane affects the cerebellar mechanisms controlling motor activity and it is postulated that this action contributes to the decrease in muscle tone induced by enflurane. PMID- 6295422 TI - Small-cell lung cancer: initial treatment with sequential hemi-body irradiation VS 3-drug systemic chemotherapy. AB - The therapeutic value of sequential hemi-body irradiation (HBI) as a primary treatment for small-cell lung cancer (SCLC) was compared to 3-drug cyclic chemotherapy (CC) in a group of 64 patients with early and advanced disease. Thirty patients were randomized to receive sequential HBI and 34 to receive CC. All patients received a local radiation boost to the primary lesion. An overall response rate of 87% was obtained in patients treated with sequential HBI and 88% in patients treated with CC. In patients with early disease, the estimated median survival was 43 weeks when treated with HBI and 42 weeks when treated with CC, but in advanced disease the estimated median survival was 15 weeks and 44 weeks respectively. Of the patients with an initial complete response, the estimated median survival was 51 weeks for HBI and 62 weeks for CC. From these observations we suggest that sequential HBI treatment technique with local radiation boost is an efficient method of tumour control in patients with early small-cell lung cancer. PMID- 6295423 TI - Correlation of collagenase secretion with metastatic-colonization potential in naturally occurring murine mammary tumours. AB - We report evidence for the secretion of a true mammalian collagenase active against Type 1 collagen, by naturally-occurring mammary tumours of the mouse and show that tumours capable of heavily colonizing the lungs secrete significantly more of this enzyme than those with low pulmonary-colonization potential, or non neoplastic proliferating (e.g. lactating) mammary tissue. Plasminogen activator is secreted in greater quantity by tumours than by normal tissues, but there is no significant difference in the amount produced by tumours with high or low pulmonary-colonization potential. These findings correlate well with our earlier morphological observations of marked connective tissue destruction in the vicinity of invading tumours and metastatic deposits, and indicate that protease release is implicated in the mechanism of tumour spread. PMID- 6295424 TI - Cell aggregates in the soft agar "human tumour stem-cell assay". AB - We evaluated colony formation in soft agar by cells obtained after mechanical and/or enzymatic disaggregation of 455 malignant human tumours. Counting and assessment of cell colonies in the agar plates were done by inverted microscopy, computerized image analysis, and inspection of serial photomicrographs of the agar plates. Our results indicate that standard methods of tumour disaggregation did not usually produce single-cell suspensions and that aggregates of tumour cells varying greatly in size were placed in the agar. Most groupings of cells identified as colonies 1-3 weeks after plating arose from enlargement of preexisting aggregates of cells. PMID- 6295426 TI - Potentiation by the human liver fluke, Opisthorchis viverrini, of the carcinogenic action of N-nitrosodimethylamine upon the biliary epithelium of the hamster. PMID- 6295425 TI - Prostaglandins and human lung carcinomas. AB - Lung primary carcinomas and normal tissue from 136 patients have been extracted for prostaglandins, and the findings examined in relation to histology. In most cases, tumours yielded more prostaglandin-like material (PG-lm), as judged by bioassay, than did normal tissue from the same lungs. Amounts varied with tumour types, in the following ascending order: small-cell carcinomas, large-cell undifferentiated carcinomas, well-differentiated squamous carcinomas, poorly differentiated adenocarcinomas, poorly differentiated squamous carcinomas, and well-differentiated adenocarcinomas. Tumour PG-lm was highest when necrosis or the neutrophil content of the tumours were moderate, whereas PG-lm from normal lung tissue correlated with the number of macrophages. Chromatography indicated the presence of various prostaglandins, in agreement with our recent findings using gas chromatography--mass spectrometry. PMID- 6295427 TI - Survival of histologically proven carcinoma of the lung registered in the North West Thames Region, 1975-1979. PMID- 6295428 TI - Modification by WR 2721 of the response to chemotherapy of tumours and normal tissues in the mouse. AB - The sulphydryl compound WR 2721 has been combined with a range of cytotoxic drugs in the mouse and the effects upon tumours and normal tissues determined. In the acute lethality (LD50/30) assay, mean protection factors produced by WR 2721 (200 or 400 mg kg-1) were generally less than 1.3 for cyclophosphamide (CTX), CCNU and chlorambucil (CHL) but a protection factor of 1.7 was obtained for cisplatinum (cis-P) in combination with 400 mg kg-1 of WR 2721. No protection against the depression of peripheral white cell count seen at 3 days after CTX, CCNU or cis-P was obtained with either 200 or 400 mg kg-1 of WR 2721. Significant protection of the RIF-1 sarcoma by WR 2721 against CTX and cis-P induced growth delay was seen. In the KHT sarcoma, WR 2721 produced small reductions in the growth delay caused by CCNU, melphalan and CHL but these were not statistically significant. These data show less differential normal tissue protection by WR 2721 than do a number of reports in the literature. PMID- 6295431 TI - Lysosomal enzyme changes in macrophages from mice given myocrisin and infected with avirulent Semliki Forest virus. AB - Myocrisin given to mice i.p. causes depression of lysosomal enzyme activity (acid phosphatase, beta-glucuronidase and N-acetyl-beta-D-glucosaminidase) in peritoneal macrophages. If avirulent Semliki Forest virus (SFV) is given i.p. 3 h after the Myocrisin, further depression of lysosomal enzyme activity occurs, a very high titre of virus is produced in these macrophages and the virus becomes lethal, causing 100% mortality. The possible interrelationships between depressed lysosomal activity, high virus titres and the production of a lethal virus infection are discussed. PMID- 6295429 TI - Interaction of misonidazole and WR-2721--II. Modification of tumour radiosensitization. AB - Two types of mouse tumour have been used to study the radiomodifying actions of Misonidazole (MISO) and WR-2721 when used alone and in combination with each other. Single dose studies were performed in both of the tumours and fractionated studies were performed on the anaplastic carcinoma, CA MT. Radioprotection with WR-2721 was seen in both tumours, being most marked at low X-ray doses. The protection was more obvious and the sensitization by MISO less in the fractionated experiment. The combination of MISO and WR-2721 gave an intermediate response compared with either drug used alone, resulting in some sensitization with single doses and an overall protection with repeated small doses. An interactive toxicity of the 2 drugs was also observed, suggesting an additive effect when assessed in terms of lethality. These studies indicate that the effects of both MISO and WR-2721 are dependent upon the oxygen status of the cells in the tumour, and that MISO can act in an oxygen-mimetic manner to modify the radioprotection observed with WR-2721. PMID- 6295430 TI - Immunogenicity and protective efficacy in a rhesus monkey model of vaccine Ac NFUi(S-) MRC against primary type 2 herpes simplex virus infection. PMID- 6295432 TI - Activity of glucose-6-phosphatase in regenerating tubular epithelium in rat kidney after necrosis induced with mercuric chloride: a light and electronmicroscopical study. AB - The activity of glucose-6-phosphatase was investigated in the renal proximal tubules of male albino rats after necrosis induced with mercuric chloride. Between the 3rd and 14th days an active zone could be observed in the inner cortex, which in the control animals was inactive. As regeneration progressed, the number of regenerating active tubules in the cortex increased, and in the 4th week activity approached that of the controls. Ultrastructurally, the enzyme was localized in the endoplasmic reticulum and nuclear envelope and sometimes even in necrotic cells. It appeared again in the regenerating cells on the 4th day. Our histochemical results complement the published biochemical results on the analysis of tissue homogenates localizing the changes of enzyme activity. From this comparison we conclude that glucose-6-phosphatase may take part in either glucose degradation or gluconeogenesis, but whereas gluconeogenesis also occurs in the first part of the proximal tubules in the normal kidney, the enzyme may play a role in glucose consumption only under pathological conditions. PMID- 6295433 TI - Therapeutic effect of acyclovir (ZoviraxTM) on the pathogenesis of chronic murine cytomegalovirus infection in the immunodeficient nude mouse. AB - The ability of acyclovir (ZoviraxTM) to influence the outcome of chronic cytomegalovirus infection in the nude mouse has been studied over a period of 3 weeks. Nude mice infected with a low dose of murine cytomegalovirus develop a chronic progressive disease which is characterized by typical lesions in the lung, the liver, salivary glands and many other organs. After 18-21 days most nude mice die of pneumonia and hepatitis. When acyclovir (ZoviraxTM) was administered s.c. twice daily, starting 1 day after the infectious dose of virus, a marked improvement was noted. No nude mice given the drug died in the 21-day period and the number of lesions to the lung, liver and salivary glands was reduced quantitatively by a highly significant amount. PMID- 6295434 TI - Coxsackievirus B4 nephritis in the squirrel monkey. AB - Seventeen squirrel monkeys (Saimiri sciureus) were experimentally infected with Coxsackievirus B4, and the kidneys, as well as other organs, were studied for pathological changes induced by the virus. Seven (41%) of these monkeys developed renal lesions--interstitial and glomerular. The Coxsackievirus was identified in 4 of these 7 monkeys (by isolation from the renal tissue in 2, by immunofluorescence staining of viral antigen in 1, and by electron microscopic finding of viral particles in 1). The renal lesions produced by Coxsackieviral infection described in this report resemble those seen in renal disease in man. These findings support the concept that viruses can produce glomerular and interstitial renal disease. This report also describes a good animal model for the study of viral disease of the kidney. PMID- 6295435 TI - Eccrine hidradenocarcinoma of the vulva with Paget's disease. Case report with a review of the literature. PMID- 6295436 TI - Adenovirus serotypes isolated from ocular infections in London. AB - During the period 1973-8 700 adenoviruses were isolated from the eyes of patients presenting at Moorfields Eye Hospital. Of these, 678 were serotyped by a neutralisation test. Twenty-one different serotypes were identified. Serotype 3, 7, and 10 accounted for 68% of the isolates, 4, 8, 15 (15/29), and 19 for 25%, and the other 14 serotypes for 7%. Community outbreaks of ocular infections by adenovirus 3, 4, 7, 10, and 15 (15/29) were observed. Outbreaks with adenovirus 3, 7, and 10 appeared to continue for 2 years or more, whereas outbreaks with 4 and 15 (15/29) were restricted to one year or less. Hospital outbreaks by adenovirus 8 and 19 were also recorded. During the same period 18 of these 21 adenovirus serotypes were isolated from the nonocular sites (mainly respiratory tract) in 7804 cases. There was a close association in the distribution of adenovirus 1, 2, 3, 5, and 6 in the ocular and nonocular sites. No such association was observed for adenovirus serotypes 4, 8, 10, 15 (15/29), and 19. PMID- 6295437 TI - Adaptive increase in phytate digestibility by phosphorus-deprived rats and the relationship of intestinal phytase (EC 3.1.3.8) and alkaline phosphatase (EC 3.1.3.1) to phytate utilization. AB - 1. The effects of phosphorus deprivation on phytate digestibility, phosphorus utilization and intestinal phytase (EC 3.1.3.8) and alkaline phosphatase (EC 3.1.3.1) in rats were investigated. 2. P deprivation was achieved by giving rats a diet containing 3 g P/kg and resulted in hypophosphataemia, hypercalcaemia, hypercalciuria, and lower levels of P absorbed and retained, and calcium retained. 3. Rats adapted to P deprivation by increasing the digestion of total dietary-P and phytate-P. 4. Levels of intestinal alkaline phosphatase and alkaline phytase were not different between the two treatment groups. 5. P deprivation in the rats given the marginal-P diet may be a result of a lower absorption of total dietary-P or increased absorption of inositol phosphates formed during the enzymatic hydrolysis of phytate which are not readily utilized by the rat. 6. These results suggest that intestinal phytase and alkaline phosphatase do not play a role in the adaptive increase in phytate digestibility by rats given marginal-P diets. The adaptation may result from enhanced phytase or alkaline phosphatase synthesis by the gastrointestinal microflora stimulated by a lower level of P in the digesta. PMID- 6295438 TI - Inhibition of angiotensin converting enzyme by phosphoramidates and polyphosphates. AB - N alpha-Phosphoryl-L-alanyl-L-proline is a reversible competitive inhibitor of angiotensin converting enzyme with a Ki of 1.4 nM. Alkylation of one phosphate oxygen with methyl, ethyl, or benzyl does not change the Ki. The high activity of the O-alkylated inhibitors demonstrates that the two phosphate oxygen anions do not constitute a bidentate ligand of the active site zinc ion. Substitution of valyltryptophan, glycylglycine, or delta-aminovaleric acid for alanylproline in the phosphoramidate raises the Ki to 12 nM, 25 microM, and 178 microM, respectively. Methylation of the alanine nitrogen in phosphorylalanylproline raises the Ki to 29 microM. Polyphosphates inhibit converting enzyme with the following Ki's: phosphate, approximately 300 mM; pyrophosphate, 2 mM; tripolyphosphate, 18 microM; tetrapolyphosphate, 150 microM. The inhibition by tripolyphosphate appears to be competitive and is unaffected by the addition of excess zinc ion. Since the Ki of tripolyphosphate is nearly 10-fold lower than that of N-phosphoryl-delta-aminovaleric acid and is near that of N alpha phosphorylglycylglycine, its terminal phosphates may bind the zinc site and the cationic site on the enzyme, thus spanning the S1' and S2' sites. PMID- 6295439 TI - Proteolipid of adenosinetriphosphatase from yeast mitochondria forms proton selective channels in planar lipid bilayers. AB - Proteolipid isolated from yeast mitochondrial adenosinetriphosphatase by butanol extraction is reincorporated into lipid vesicles from which planar membranes are formed. The proteolipid permits electric conductance through the membrane. This conductance occurs through membrane channels which are highly selective for protons. Proton channels in the membrane are directly observed at high proton concentrations in the aqueous phases. Channels open and close independently from each other; their open-state conductances and lifetimes are monodisperse but influenced by the applied voltage (12 pS and 3 s, respectively, at pH 2.2 and 100 mV). Proton channels do not occur in single proteolipid molecules; the conducting structure consists of at least two polypeptide chains since channels form in a (reversible) bimolecular reaction of nonconducting forms of proteolipid. The number of proton channels at a constant proteolipid concentration changes in sharp transitions and by orders of magnitudes upon critical changes of membrane composition and pH. These transitions are caused by transitions of proteolipid organization in the membrane from a dispersed state (equilibrium between channel forming "dimers" and a large pool of "monomers") to a state of almost complete aggregation of proteolipid which stabilizes large proton-conducting structures (probably associates of channel-forming dimers). This self-association of isolated proteolipid into structures containing proton-selective channels suggests that the six proteolipids in the adenosinetriphosphatase complex exist as a self-associating entity containing most likely three proton channels. PMID- 6295440 TI - Adenosine cyclic 3',5'-monophosphate dependent protein kinase: kinetic mechanism for the bovine skeletal muscle catalytic subunit. PMID- 6295441 TI - Mechanism of lactose transport in Escherichia coli membrane vesicles: evidence for the involvement of histidine residue(s) in the response of the lac carrier to the proton electrochemical gradient. PMID- 6295442 TI - Direct measurement of lactose/proton symport in Escherichia coli membrane vesicles: further evidence for the involvement of histidine residue(s). AB - Addition of lactose to Escherichia coli ML 308-225 membrane vesicles under nonenergized conditions induces transient alkalinization of the medium, and the initial rate of proton influx is stimulated by valinomycin and abolished by nigericin or carbonyl cyanide m-chlorophenylhydrazone. A functional lac y gene product is absolutely required as the effect is not observed in ML 308-225 vesicles treated with N-ethylmaleimide nor with vesicles from uninduced Escherichia coli ML 30. Furthermore, the magnitude of the phenomenon is enhanced about 3-fold in vesicles from Escherichia coli T206, which contain amplified levels of the lac carrier protein. Kinetic parameters for lactose-induced proton influx are the same as those determined for lactose-facilitated diffusion, and quantitative comparison of the initial rates of the two fluxes indicates that the stoichiometry between protons and lactose is 1:1. Treatment of ML 308-225 vesicles with diethyl pyrocarbonate causes inactivation of lactose-induced proton influx. Remarkably, however, treatment with the histidine reagent enhances the rate of lactose-facilitated diffusion in a manner suggesting that the altered lac carrier catalyzes lactose influx without the symport of protons. The results are consistent with the hypothesis that acylation of a histidyl residue(s) in the lac carrier protein dissociates lactose influx from proton influx and indicate that this residue(s) play(s) an important role in the pathway of proton translocation. PMID- 6295443 TI - Phage P22 tail protein: gene and amino acid sequence. AB - The tail structure of the Salmonella phage P22 mediates both adsorption of the phage to its host and enzymatic hydrolysis of the bacterial O-antigen. The tail is an oligomeric structure, which is assembled from a single polypeptide species. We report here the amino- and carboxyl-terminal sequences of the P22 tail protein and the nucleotide sequence of its gene (gene 9). These data specify the complete amino acid sequence of the tail protein. The tail protein is a slightly acidic protein containing 666 amino acids. Comparison of the gene and protein sequences indicates that mature tail protein arises by cleavage of the initiator N-formyl methionine from the nascent chain. PMID- 6295444 TI - Proton nuclear magnetic resonance studies of Pseudomonas testosteroni 3-oxo-delta 5-steroid isomerase and its interaction with 17 beta-estradiol. PMID- 6295445 TI - Phosphorus-31 nuclear magnetic resonance studies of the two phosphoserine residues of hen egg white ovalbumin. AB - Ovalbumin contains two phosphoserine residues that give rise to two well-resolved resonances in a 31P NMR spectrum. Ovalbumin samples that have been digested with a variety of phosphatases may give rise to only one phosphoserine resonance, indicating that one of the two phosphorylated sites is relatively inaccessible for phosphatase action. By comparison of the amino acid sequence of the peptide containing the nonsusceptible phosphate to the overall primary structure, we have assigned the resonances observed (pH 8.3) at 5.0 and 4.75 ppm to phosphoserines 68 and -344, respectively. pH titration behavior and susceptibility of the phosphoserine residues to phosphatases indicate that both are located on the surface of the protein. Both residues have a pKa = 6.00-6.04. Analysis of the Hill coefficients measured for the pH titrations and the JPH coupling constants indicate that neither residue interacts with other charged groups on the surface of the protein. Frequency dependence of 31P NMR parameters shows that at higher magnetic field strengths the contribution of chemical shift anisotropy to the line width becomes very significant. We have calculated from the field-dependent terms that phosphoserine-344 is mobile with respect to the protein surface but that phosphoserine-68 is more restricted in its motion. The latter is also involved in a pH-dependent conformational change, since it is shielded from hydrolysis by phosphatases at higher pH. A comparison of the amino acid sequence of the phosphoserine-68 site shows that it has a striking homology to the active site peptides of a wide variety of hydrolytic enzymes. Moreover, a comparison with the primary sequences of casein suggests that both proteins are phosphorylated by a protein kinase that specifically recognizes a Ser-X-Glu peptide. PMID- 6295446 TI - Bile pigment--protein interactions. Coupled oxidation of cytochrome c. AB - A new methodology is described for the chemical modification of the heme prosthetic group of horse heart cytochrome c. The selective modification of the heme moiety of cytochrome c is facilitated by utilizing coupling oxidation conditions. Comparison of the absorption spectra of this chemically modified cytochrome c species in two different solvents (aqueous pyridine and carbon monoxide saturated 6 M guanidinium chloride) with those of two model compounds [bis(pyridine)(2,3,7,8,12,13,17,18-octaethyl-5-oxaporphyrinato)iron(II) tetrafluoroborate salt and (pyridine)carbonyl-(2,3,7,8,12,13,17,18-octaethyl-5 oxaporphyrinato)iron(II) tetrafluoroborate salt] shows that coupled oxidation of cytochrome c affords a new protein with a covalently bound iron(II) oxaporphyrin prosthetic group. Amino acid analysis of this protein-bound iron(II) oxaporphyrin species reveals that only limited modification of the primary structure of the apoprotein occurs during coupled oxidation of cytochrome c. This protein-bound iron(II) oxaporphyrin species is also interconvertible to a protein-bound bilatriene species under hydrolytic conditions. The synthetic utility of the coupled oxidation of cytochrome c for the preparation of chromoproteins which possess covalently bound iron(II) oxaporphyrin and bilatriene prosthetic groups is considered. PMID- 6295447 TI - Spin-label saturation-transfer electron spin resonance detection of transient association of rhodopsin in reconstituted membranes. AB - Rotational diffusion of rhodopsin in reconstituted membranes of phosphatidylcholines of various alkyl chain lengths has been measured by using saturation-transfer electron spin resonance spectroscopy as a function of temperature and lipid/rhodopsin mole ratio. For dipalmitoyl-phosphatidylcholine, the rotational correlation time is 20 microseconds at physiological concentration, the same as in rod outer segment (ros) membranes. Dilution reduces the time to 10 microseconds, a value that is ascribed to well-dispersed monomeric rhodopsin. Use of phospholipids with longer or shorter chains results in sharply increased rotational correlation times. It is concluded that rhodopsin molecules are transiently associated in both reconstituted and ros membranes and that the nature of the association is determined by lipid type and composition. PMID- 6295448 TI - Exclusive labeling of the extracytoplasmic surface of sodium ion and potassium ion activated adenosinetriphosphatase and a determination of the distribution of surface area across the bilayer. PMID- 6295449 TI - Numbers and exchangeability with water of oxygen-17 atoms coupled to molybdenum (V) in different reduced forms of xanthine oxidase. AB - The effect of using [17O]water (24-50% enriched) as solvent on the Mo(V) electron paramagnetic resonance spectra of different reduced forms of xanthine oxidase has been investigated. All the Mo(V) signals are affected. Procedures are described, based on the use of difference spectral techniques, that facilitate interpretation of such spectra. The number of coupled oxygen atoms may be determined by estimation of the fraction of the spectrum that remains unchanged by the isotope at a known enrichment. For a species having two coupled oxygen atoms, the use of two different isotope enrichments permits elimination from the difference spectra of the contribution of the two singly substituted species. From the application of these methods, it is concluded that not only the strength of the hyperfine coupling of oxygen ligands of molybdenum but also their number and their exchangeability with the solvent vary from one reduced form of the enzyme to another. The inhibited species from active xanthine oxidase has been studied in the most detail. It has two weakly coupled oxygen atoms [A(17O)av = 0.1-0.2 mT] that do not exchange with the solvent. A cyclic structure is proposed for this species in which two oxygen ligands of molybdenum are bonded to the carbon of the formaldehyde or other alcohol or aldehyde molecule that reacted in producing the signal. Structures of the other signal-giving species from active xanthine oxidase (Very Rapid and Rapid types 1 and 2) are discussed, as is corresponding information on species from the desulfo enzyme and from sulfite oxidase. PMID- 6295450 TI - Specific stimulation of the T7 gene 6 exonuclease by the phage T7 coded deoxyribonucleic acid binding protein. AB - Bacteriophage T7 codes for a single-stranded DNA binding protein. This protein is the product of gene 2.5 and has been found previously to stimulate specifically the activity of the phage-coded DNA polymerase. We report here that the T7 DNA binding protein also stimulates the activity of the phage-coded exonuclease. The gene 6 exonuclease is a double-stranded DNA specific 5'-exonuclease that has been implicated in destruction of bacterial DNA, removal of RNA primers during DNA replication, genetic recombination, and DNA maturation. The enzyme is markedly inhibited by physiological concentrations of NaCl. This inhibition, which is due to a marked reduction in the Vmax of the enzyme, can be largely overcome by the phage-coded DNA binding protein. This stimulation is specific since the Escherichia coli DNA binding protein is without effect. The stimulation by the binding protein is apparently not due to its coating of the 3' single-stranded tails generated during the digestion. Kinetic studies show that the stimulation is due to a combined effect on both the Km and Vmax of the exonuclease. These studies are consistent with a loose binding of the binding protein to either the DNA or the exonuclease. PMID- 6295451 TI - Purification of human neutrophil collagenase and production of a monospecific antiserum. AB - Although there is good evidence for the presence of human neutrophil (PMN) collagenase, only moderate purification has been reported. The probable explanation for this fact is that most assays used to specifically measure collagenase activity are not reliable if high levels of several different proteases are also present in the assay mixture. The PMN granule is just such a concentrated mixture. Therefore, polyacrylamide gel electrophoresis was used to identify and quantitate the alpha 1 3/4 and alpha 2 3/4 cleavage products diagnostic for mammalian collagenase. White cells (85% PMN's) were lysed in 0.34 M sucrose and granules were obtained. The granules were lysed by sonication, and the lysate was chromatographed on a Sephadex G-200 column followed by a Trasylol Sepharose 4B column. This procedure resulted in a 1350-fold purification and a yield of 75 micrograms of enzyme/unit of blood. The collagenase was inhibited by ethylene glycol bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid but not by sulfhydryl or serine protease inhibitors. The preparation was free of elastase, which has been shown to cleave type III collagen into alpha 1 3/4 and alpha 1 1/4 pieces. The pI of collagenase was shown to be 4.7 by isoelectric focusing, and the enzyme lost activity below a pH of 6.5 if collagen was absent. Antiserum was produced by 100-micrograms injections of the purified collagenase into rabbits. Titers were measured by the enzyme-linked immunosorbent assay. For determination of the specificity, collagenase and PMN extract were isoelectrically focused and blotted onto nitrocellulose. The antibody recognized only one band of protein in the PMN extract, which comigrated with the purified collagenase. PMID- 6295452 TI - Stoichiometry of catabolite activator protein/adenosine cyclic 3',5' monophosphate interactions at the lac promoter of Escherichia coli. PMID- 6295453 TI - Sequence analysis of a growth hormone releasing factor from a human pancreatic islet tumor. AB - A growth hormone releasing factor of a human pancreatic islet tumor (hpGRF) of an acromegalic patient was purified and subjected to Edman degradation in a spinning cup sequencer. Approximately 0.7-1.2 nmol of peptide was applied to the cup without any pretreatment, after coupling to 3-sulfophenyl isothiocyanate or after cleavage with cyanogen bromide, staphylococcal protease, or trypsin. On the basis of the analytical data, the N-terminal sequence of 39 residues is established to be H-Tyr-Ala-Asp-Ala-Ile-Phe-Thr-Asn- Ser-Tyr-Arg-Lys-Val-Leu-Gly-Gln-Leu-Ser-Ala Arg-Lys- Leu-Leu-Gln-Asp-Ile-Met-Ser-Arg-Gln-Gln-Gly-Glu-Ser- Asn-Gln-Glu-Arg-Gly . It is proposed that alanine is residue 40 and represents (as free acid) the C terminus of hpGRF. Synthetic hpGRF(1-40)-OH is highly potent in stimulating GH secretion from the rat anterior pituitary in vitro and in vivo. The C-terminal sequence of hpGRF does not appear to contribute significantly to the biologic intrinsic activity and potency of hpGRF, as demonstrated by the fact that the natural product and the synthetic peptides hpGRF(1-40)-OH, hpGRF(1-40)-NH2, and hpGRF(1-29)-NH2 show equivalent in vitro activities. On the basis of sequence homologies, hpGRF is closely related to members of the glucagon secretin family, especially to the porcine gut peptide PHI. PMID- 6295454 TI - Sendai virus membrane fusion: time course and effect of temperature, pH, calcium, and receptor concentration. AB - The conditions that optimize Sendai virus membrane fusion with liposomes have been studied. No fusion occurs in the absence of ganglioside receptors. Maximum fusion occurs when the molar ratio of ganglioside GD1a to phospholipid is 0.02 or greater. The amount of fusion at 37 degrees C increases with time up to at least 6.5 h. The rate of fusion increases from the lowest temperature tested, 10 degrees C, to 40 degrees C. Above 43 degrees C the amount of fusion decreases because of thermal inactivation of the viral proteins. There is a broad pH maximum between pH 7.5 and pH 9.0. At both ends of the pH range the amount of fusion increases and exceeds that found in the physiologic pH range. Neither ethylenediaminetetraacetic acid nor Ca2+ changes the amount of membrane fusion. The optimal conditions for membrane fusion of Sendai virus membranes with liposomes are the same as the optimal conditions for fusion with host cells and with red blood cells. Since the liposomes contain no proteins, the optimal conditions for Sendai virus membrane fusion must be determined by the viral proteins and be mostly independent of the nature or presence of the host proteins. PMID- 6295456 TI - Isolation and characterization of diadenosine 5',5"'-P1,P4-tetraphosphate pyrophosphohydrolase from Physarum polycephalum. AB - A new enzyme that hydrolyzes diadenosine 5',5"'-P1,P4-tetraphosphate has been purified by a factor of 250 from the acellular slime mold Physarum polycephalum. Activity was assayed radioisotopically with [3H]Ap4A. Isolation of the enzyme was facilitated by dye-ligand chromatography. The enzyme symmetrically hydrolyzes Ap4A to ADP and exhibits biphasic kinetics for the substrate with values for the apparent Km of 2.6 micro M and 37 micro M. The two values of Vmax differ by a factor of 10. Mg2+, Ca2+, and other divalent cations inhibit the activity with 40 80% inhibition occurring at 0.5 mM. Mg2+, at 0.5 mM, decreases both values of Vmax by 50%, decreases the low Km value by about 30%, and increases the high Km value by about 100%. (Ethylenedinitrilo)tetraacetic acid (EDTA) and [ethylenebis(oxyethylenenitrilo)]tetraacetic acid (EGTA), at 10 mM, inhibit the activity by 50%. ADP, ATP, Ap4, and Gp4 are equipotent inhibitors with 50% inhibition occurring at 30 micro M. AMP is a relatively weak inhibitor. The molecular weight of the enzyme is 26000 on the basis of elution of activity from a calibrated Sephadex G-75 column. PMID- 6295455 TI - Nuclear magnetic resonance studies of the nucleotide binding sites of porcine adenylate kinase. AB - The alpha, beta, gamma-tridentate complex of CrATP, a paramagnetic competitive inhibitor of porcine adenylate kinase, increases the longitudinal [1/(fT1p)] and transverse [1/(fT2p)] relaxation rates of a resonance of the enzyme previously assigned by McDonald et al. [McDonald, G.G., Cohn, M., & Noda, L. (1975) J. Biol. Chem. 250, 6947-6954] to the C2 proton of histidine-36. These paramagnetic effects are diminished upon the addition of the substrate MgATP by an amount consistent with the simple displacement of CrATP. The 1/(fT2p) value sets a lower limit of 400 s-1 on the rate constant for dissociation of CrATP from the enzyme. The 1/(fT1p) value at 250 MHz and the correlation time for water protons in the same complex are used to calculate a distance of 12.9 +/- 1.0 A from Cr(III) to the C2 proton of histidine-36. A primary, negative nuclear Overhauser effect is detected on the adenine H2 resonance of enzyme-bound MgATP upon preirradiation of the C2 proton of histidine-36, indicating that these protons are approximately less than 5 A apart. These distances and negative intramolecular Overhauser effects from the ribose protons to adenine H8 of MgATP indicate an extended structure for bound MgATP with an anti conformation about the glycosidic bond. These findings require a different orientation or location of the bound metal-ATP substrate from that proposed based on X-ray studies of the binding of salicylate [Pai, E. F., Sachsenheimer, W., & Schirmer, R.H. (1977) J. Mol. Biol. 114, 37 45]. Other nuclear Overhauser effects from resonances of the protein at 1.8 and 0.9 ppm to both adenine H2 and ribose H1' of bound MgATP indicate the proximity to the substrate of at least one Arg C beta proton (at 1.8 ppm), C gamma proton (at 1.7 ppm), Lys C delta proton (at 1.7 ppm), or Leu C beta proton (at 1.6 ppm) and one or more Leu, Ile, or Val methyl groups (at 0.9 ppm). Entirely different Overhauser effects are observed from the enzyme to the adenine protons of AMP consistent with a distinct site for the other substrate. PMID- 6295457 TI - Diadenosine 5',5"'-P1,P4-tetraphosphate pyrophosphohydrolase from Physarum polycephalum. Substrate specificity. AB - The substrate specificity of diadenosine 5',5"'-P1,P4-tetraphosphate pyrophosphohydrolase from Physarum polycephalum for dinucleoside polyphosphates has been determined by high-performance liquid chromatography (HP-LC). Elution of a strong anion-exchange resin with a pH and ionic strength gradient of ammonium phosphate separates a series of monoadenosine and diadenosine polyphosphates. Most of the corresponding guanine nucleotides are also resolved on this HPLC system. One mole each of Ap4A and Gp4G is symmetrically hydrolyzed to 2 mol of ADP and GDP, respectively. Ap3A, Ap5A, Ap6A, and Ap4 are hydrolyzed, and in each case ADP is one of the products. Gp3G, Gp5G, Gp6G, and Gp4 are also substrates, and in each case GDP is one of the products. AMP, ADP, ATP, Ap2A, ADPR, GMP, GDP, GTP, NAD+, and NADP+ are not substrates. No hydrolysis of the cap dinucleotides m7Gp3Am and m7Gp3Cm was detected by HPLC. Diadenosine tetraphosphate pyrophosphohydrolase preparations were also assayed for adenylate kinase, nucleotide diphosphate kinase, NAD(P)+ pyrophosphohydrolase, phosphodiesterase, cyclic nucleotide phosphodiesterase, phosphatase, and ribonuclease activities. These enzymic activities were not detectable in diadenosine tetraphosphate pyrophosphohydrolase. The symmetrical hydrolysis of Ap4A and Gp4G is an unique catalytic property that distinguishes diadenosine tetraphosphate pyrophosphohydrolase from P. polycephalum from diadenosine tetraphosphate phosphohydrolases from other organisms. PMID- 6295458 TI - Molecular aspects of bovine erythrocyte bis[(heme b)copper] protein. AB - An erythrocyte Cu2(heme b)2 protein of Mr 400000 was successfully isolated. Incubating the protein in sodium dodecyl sulfate prior to polyacrylamide gel electrophoresis caused the splitting into Mr 70000, 120000, and 200000 units. The copper was fully electron paramagnetic resonance detectable of the type II (g perpendicular = 2.0309, g parallel = 2.2122, A parallel = 175 G). The high-spin (d55/2) iron(III) showed a g value of 6.05. No magnetic interaction between copper and heme iron was detected. In a comparison of more than ten different enzymatic oxidase activities, not a single one could be assigned to the Cu2(heme b)2 protein. Azide, CO, cyanide, fluoride, and imidazole were bound to the heme iron. The binding of imidazolate suggests the accessibility of the sixth coordination site of the heme b group to fairly large ligands. Removal of the copper by ethylenediaminetetraacetic acid or cyanide resulted in an irreversible precipitation of the protein. This supports the structural contribution of the copper. PMID- 6295459 TI - Inhibition of tyrosine protein kinases by halomethyl ketones. AB - A chloromethyl ketone derivative of lactic acid was shown to inhibit protein phosphorylation in plasma membranes of Ehrlich ascites tumor cells [Johnson, H. J., Zimniak, A., & Racker, E. (1982) Biochemistry 21, 2984-2989]. We now show that this inhibitor as well as three halomethyl ketone derivatives of amino acids and peptides specifically inhibits tyrosine protein kinase activity in intact plasma membranes and Triton extracts of plasma membrane of A-431 tumor cells. The most effective inhibitor is a bromomethyl ketone derivative of leucine that inhibits the phosphorylation of a protein that migrates to the same position as the EGF receptor in sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Inhibition of phosphorylation took place in the presence or absence of added EGF, and the inhibitor did not interfere with the binding of EGF to the receptor nor with the dephosphorylation of the EGF-stimulated phosphoprotein. EGF-dependent phosphorylation in a Triton extract of plasma membranes from normal placenta was considerably less sensitive to the bromomethyl ketone derivative of leucine. The tyrosine protein kinase activity of the transformation gene product of Fujinami virus was particularly sensitive to the bromomethyl ketone derivative of leucine, while the src gene product of Rous sarcoma virus was comparatively less sensitive. The bromomethyl ketone inhibitor interfered with the phosphorylation of the EGF receptor by [gamma-32P]-8-azido-ATP but much less with the light sensitive binding. This observation and the lack of interference with EGF binding suggest that the inhibitor interacts with the protein kinase portion of the receptor complex. PMID- 6295460 TI - Biochemical characterization of the tetrodotoxin binding protein from Electrophorus electricus. AB - Biochemical properties of a detergent-solubilized tetrodotoxin binding component from Electrophorus electricus have been examined and compared with those found for the membrane-bound protein. The toxin binding component was solubilized with high efficiency by a variety of nonionic detergents and with lower efficiency by sodium cholate and deoxycholate. Detergent-solubilized preparations bound tetrodotoxin and saxitoxin tightly and specifically, and this binding was observed to be rapidly and irreversibly blocked by carboxylate-modifying reagents. Inactivation by carbodiimide and glycine ester or by a trimethyloxonium salt could be prevented by tetrodotoxin occupancy of the binding site. Tetrodotoxin binding activity in both solubilized preparations and in membranes was found to be highly resistant to proteases. In contrast, the activity was extremely sensitive to the action of phospholipase A2. The biochemical properties of the tetrodotoxin binding component solubilized in mixed lipid-detergent micelles are similar to those found in native membranes, with respect to the characteristics of equilibrium toxin binding and to the sensitivity of toxin binding activity to chemical modification and degradative enzymes. There were some differences with respect to the kinetics of tetrodotoxin binding. In addition, the tetrodotoxin binding component from eel is shown to behave as a glycoprotein, being selectively absorbed to resins coupled to concanavalin A, wheat germ agglutinin, Lens culinaris lectin, and ricin with the appropriate glycoside. PMID- 6295461 TI - Myeloperoxidase-catalyzed incorporation of amines into proteins: role of hypochlorous acid and dichloramines. AB - Myeloperoxidase-catalyzed oxidation of chloride (Cl-) to hypochlorous acid (HOCl) resulted in formation of mono- and dichloramine derivatives (RNHCl and RNCl2) of primary amines. The RNCl2 derivatives could undergo a reaction that resulted in incorporation of the R moiety into proteins. The probable mechanism was attack of RNCl2 or an intermediate formed in the decomposition of RNCl2 on histidine, tyrosine, and cystine residues and on lysine residues at high pH. Incorporation of radioactivity from labeled amines into stable, high molecular weight derivatives of proteins was measured by acid or acetone precipitation and by gel chromatography and electrophoresis. Whereas formation of RNCl2 was favored at low pH, the subsequent incorporation reaction was favored at high pH. Up to several hours were required for the maximum amount of incorporation, which was less than 10% of the label in RNCl2. For the amines tested, incorporation was in the order histamine greater than 1,2-diaminoethane greater than putrescine greater than taurine greater than lysine greater than glucosamine greater than leucine greater than methylamine. Initiation of the reaction required HOCl, and oxidized forms of bromide, iodide, or thiocyanate did not substitute. Inhibitors of incorporation fell into three classes. First, ammonia or amines competed with the labeled amine for reaction with HOCl, so that larger amounts of HOCl were required. Second, readily oxidized substances such as sulfhydryl or diketo compounds or thioethers (methionine) reduced RNCl2. Third, certain compounds competed with protein as the acceptor for the incorporation reaction. The amount required to block incorporation into protein depended on protein concentration. Among these inhibitors were imidazole compounds (histidine), phenols (tyrosine), and disulfides (glutathione disulfide, GSSG). Low yields of derivatives of histidine, tyrosine, and GSSG were detected by thin-layer chromatography. Acid-precipitable derivatives were obtained by reacting RNCl2 with polyhistidine or polytyrosine, and to a lesser extent with polylysine at high pH, but not with other poly(amino acids). Precipitable derivatives were also obtained by incubating MPO-containing extracts from leukocyte granules with hydrogen peroxide, Cl-, and labeled amines. The extracts were found to have a high content of substances with primary amino groups, which competed for incorporation. The results account for oxidative incorporation of amines into proteins in leukocytes and provide evidence that HOCl and nitrogen-chlorine (N-Cl) derivatives are formed in these cells. The characteristics of the incorporation reaction suggest that it would not contribute significantly to the antimicrobial activity of myeloperoxidase (MPO). Nevertheless, the reaction may provide a sensitive method for studying MPO action in vivo. PMID- 6295462 TI - Exploration of the nucleotide binding sites of the isolated ADP/ATP carrier protein from beef heart mitochondria. 1. Probing of the nucleotide sites by Naphthoyl-ATP, a fluorescent nontransportable analogue of ATP. AB - The ADP/ATP carrier protein was extracted and purified from beef heart mitochondria, and its binding parameters with respect to 3'-O-naphthoyladenosine 5'-triphosphate (N-ATP), a fluorescent nontransportable analogue of ATP, were studied. The binding of N-ATP to the isolated carrier protein was accompanied by a decrease in fluorescence. Conversely, the release of bound N-ATP upon addition of carboxyatractyloside (CATR) or ATP resulted in a fluorescence increase. The bound N-ATP that was released upon addition of an excess of CATR or ATP was referred to as specifically bound N-ATP, i.e., N-ATP bound to the nucleotide sites of the carrier protein. Two classes of binding sites for N-ATP could be identified; the number of high-affinity sites (Kd less than 10 nM) was equal to the number of low-affinity sites (Kd = 0.45 microM). CATR behaved apparently as a noncompetitive inhibitor of the binding of N-ATP. The amount of N-ATP released increased linearly with the amount of CATR added, indicating an extremely high affinity of the carrier protein for CATR. The number of CATR binding sites was equal to half the total number of N-ATP binding sites (high- and low-affinity sites); at saturating concentrations of N-ATP, the binding of 1 mol of CATR resulted in the release of 2 mol of bound N-ATP, one from the high-affinity site and the other from the low-affinity site, showing unambiguously that each CATR site is interacting with a pair of probably interdependent N-ATP sites. A clear competition between N-ATP and ATP for binding to the carrier protein was demonstrated. The Kd values of the high- and low-affinity sites for ATP were less than 50 nM and 5 microM, respectively. In the presence of high concentrations of ATP, the two classes of N-ATP binding sites became indistinguishable, suggesting interconversion. It is proposed that the asymmetry in affinity for N-ATP binding is induced by the binding step itself, the carrier protein exhibiting a negative cooperativity for N-ATP binding. PMID- 6295463 TI - Exploration of the nucleotide binding sites of the isolated ADP/ATP carrier protein from beef heart mitochondria. 2. Probing of the nucleotide sites by formycin triphosphate, a fluorescent transportable analogue of ATP. PMID- 6295464 TI - Transient kinetics of electron transfer reactions of flavodoxin: ionic strength dependence of semiquinone oxidation by cytochrome c, ferricyanide, and ferric ethylenediaminetetraacetic acid and computer modeling of reaction complexes. AB - Electron transfer reactions between Clostridum pasteurianum flavodoxin semiquinone and various oxidants [horse heart cytochrome c, ferricyanide, and ferric ethylenediaminetetraacetic [horse heart cytochrome c, ferricyanide, and ferric ethylenediaminetetraacetic acid (EDTA)] have been studied as a function of ionic strength by using stopped-flow spectrophotometry. The cytochrome c reaction is complicated by the existence of two cytochrome species which react at different rates and whose relative concentrations are ionic strength dependent. Only the faster of these two reactions is considered here. At low ionic strength, complex formation between cytochrome c and flavodoxin is indicated by a leveling off of the pseudo-first-order rate constant at high cytochrome c concentration. This is not observed for either ferricyanide or ferric EDTA. For cytochrome c, the rate and association constants for complex formation were found to increase with decreasing ionic strength, consistent with negative charges on flavodoxin interacting with the positively charged cytochrome electron transfer site. Both ferricyanide and ferric EDTA are negatively charged oxidants, and the rate data respond to ionic strength changes as would be predicted for reactants of the same charge sign. These results demonstrate that electrostatic interactions involving negatively charged groups are important in orienting flavodoxin with respect to oxidants during electron transfer. We have also carried out computer modeling studies of putative complexes of flavodoxin with cytochrome c and ferricyanide, which relate their structural properties to both the observed kinetic behavior and some more general features of physiological electron transfer processes. The results of this study are consistent with the ionic strength behavior described above. PMID- 6295465 TI - Purification and characteristics of a mitochondrial endonuclease from the yeast Saccharomyces cerevisiae. AB - Saccharomyces cerevisiae contains a membrane-bound mitochondrial nuclease. The enzyme was purified nearly 500-fold from sphaeroplasts of the organism by differential centrifugation, differential solubilization, heparin-agarose chromatography, and gel filtration. A final specific activity of 98 mumol min-1 (mg of protein)-1 was obtained. The enzyme required further purification to achieve homogeneity. Two peaks of activity were obtained after gel filtration with apparent molecular weights of 140000 and 57000. Otherwise, these two components have nearly identical characteristics. Without detergent the enzyme is insoluble and has very low activity. Zwittergent 3-14 or Triton X-100 in the presence of KCl could be used to solubilize and activate the enzyme. A number of other detergents were much less effective in solubilizing or activating the nuclease. The enzyme requires Mg2+ for activity, and this can be replaced to some degree by Mn2+ but not by Ca2+ or Zn2+. It is most active at pH 6.5-7.0 and degrades the substrate to small oligonucleotides with 5'-phosphate ends. The relative rates of hydrolysis were 100 for poly(A), 31 for ssDNA, 19 for RNA, 2.1 for dsDNA, and less than or equal to 0.2 for poly(C). Under the assay conditions used the enzyme appears to constitute about 90% of the total nuclease activity of the cell. The enzyme is unstable, especially at neutral and alkaline pH. PMID- 6295466 TI - Interactions between phospholipid head groups at membrane interfaces: a deuterium and phosphorus nuclear magnetic resonance and spin-label electron spin resonance study. PMID- 6295467 TI - Aerobactin-mediated utilization of transferrin iron. AB - Aerobactin and enterobactin, hydroxamate- and catechol-type siderophores, respectively, were found capable of removing iron (III) from transferrin in buffered solution. Although under these conditions aerobactin displaced the iron much more slowly than did enterobactin, the rate for the former could be accelerated by addition of pyrophosphate as mediator. Transfer of iron (III) from transferrin to aerobactin appeared to proceed via a ternary complex. Cells of Escherichia coli BN 3040 NalR iuc containing transport systems for both enterobactin and aerobactin, the genetic determinants for the latter specified on a ColV-type plasmid, took up iron from [55Fe]transferrin in minimal medium. In this case aerobactin was effective at a much lower concentration, although enterobactin still displayed superior ability to transfer the iron. In serum, however, the rate measured with aerobactin exceeded that found with enterobactin. The results indicate that aerobactin, in spite of its relatively unimpressive affinity for iron (III) as a siderophore, is nonetheless equipped with structural features or properties that enhance its ability to remove the metal ion from transferrin, especially when receptor-bearing cells of E. coli are present to act as a thermodynamic sink for the iron. These attributes of the aerobactin system of iron assimilation may account for its status as a virulence determinant in hospital isolates of E. coli. PMID- 6295468 TI - Chitin-bound protein of sarcophagid larvae: metabolism of covalently linked aromatic constituents. AB - The borate-insoluble chitin-protein complex, CB-I, from prepupal sarcophagid larvae was cleaved with chymotrypsin and trifluoromethanesulfonic acid releasing a polypeptide fragment of Mr 68 000. The intact glycoprotein was blocked at the C terminus; the N-terminal sequence of Asp-Val-Ala-His-Tyr was not homologous with seven of the borate-soluble nonglycosylated structural proteins. Bityrosine was identified as a component of the primary chain, both half-residues occupied in peptide linkages. Sclerotization initiated a decline in bityrosine coincident with the addition of soluble proteins to the tanned matrix. The chitin-protein complex also included bound peroxidase, propolyphenol oxidase, and an o-diphenol subject to oxidation on activation of the zymogen. In the course of the oxidation N termini declined in accordance with the formation of 1,4 quinonoid cross-links. PMID- 6295469 TI - Kinetics for exchange of imino protons in the d(C-G-C-G-A-A-T-T-C-G-C-G) double helix and in two similar helices that contain a G . T base pair, d(C-G-T-G-A-A-T T-C-G-C-G), and an extra adenine, d(C-G-C-A-G-A-A-T-T-C-G-C-G). AB - The relaxation lifetimes of imino protons from individual base pairs were measured in (I) a perfect helix, d(C-G-C-G-A-A-T-T-C-G-C-G), (II) this helix with a G . C base pair replaced with a G . T base pair, d(C-G-T-G-A-A-T-T-C-G-C-G), and (III) the perfect helix with an extra adenine base in a mismatch, d(C-G-C-A-G A-A-T-T-C-G-C-G). The lifetimes were measured by saturation recovery proton nuclear magnetic resonance experiments performed on the imino protons of these duplexes. The measured lifetimes of the imino protons were shown to correspond to chemical exchange lifetimes at higher temperatures and spin-lattice relaxation times at lower temperatures. Comparison of the lifetimes in these duplexes showed that the destabilizing effect of the G . T base pair in II affected the opening rate of only the nearest-neighbor base pairs. For helix III, the extra adenine affected the opening rates of all the base pairs in the helix and thus was a larger perturbation for opening of the base pairs than the G . T base pair. The temperature dependence of the exchange rates of the imino proton in the perfect helix gives values of 14-15 kcal/mol for activation energies of A . T imino protons. These relaxation rates were shown to correspond to exchange involving individual base pair opening in this helix, which means that one base-paired imino proton can exchange independent of the others. For the other two helices that contain perturbations, much larger activation energies for exchange of the imino protons were found, indicating that a cooperative transition involving exchange of at least several base pairs was the exchange mechanism of the imino protons. The effects of a perturbation in a helix on the exchange rates and the mechanisms for exchange of imino protons from oligonucleotide helices are discussed. PMID- 6295470 TI - Flexibility of end-labeled polymers from electron spin resonance line-shape analysis: 3' terminus of transfer ribonucleic acid and 5S ribonucleic acid. AB - Saccharomyces cerevisiae tRNA and 5S RNA, Escherichia coli 5S RNA, and wheat germ 5S RNA have each been specifically spin-labeled at the 3'-terminal ribose to give morpholino-spin-labeled (MSL) RNAs. Enzymatic hydrolysis with pancreatic RNase, followed by anion-exchange chromatography, confirms the site of attachment of the spin-label. Effective rotational correlation times, TB and TC, have been determined from electron spin resonance (ESR) peak heights and widths as a function of temperature for each MSL RNA, and Arrhenius plots of -log T vs. 1/T have been constructed. TC is a measure of internal flexibility at the link between the label and the RNA, while TB is a measure of rotational flexibility of the RNA near the labeled site. Validity of the TB and TC determination has been confirmed from simulation of the experimental EPR spectra by theoretical spectra computed for various attachment geometries and motional rates. Discontinuities in the slope of Arrhenius plots for TB were seen at 34 and 66 degrees C (yeast MSL tRNA), 37 and 60 degrees C (E. coli MSL 5S RNA), 37 and 57 degrees C (yeast MSL 5S RNA), and 36 and 54 degrees C (wheat germ MSL 5S RNA). Temperature-induced hydrolysis of each MSL RNA was less than 5% as determined by gel-filtration chromatography. The melting curves are consistent with a recently proposed universal secondary structural model for prokaryotic and eukaryotic 5S RNA. PMID- 6295471 TI - Multiphasic oxidation-reduction of cytochrome b in the succinate-cytochrome c reductase. AB - The triphasic course previously reported for the reduction of cytochrome b in the succinate-cytochrome c reductase by either succinate or duroquinol has been shown to be dependent on the redox state of the enzyme preparation. Prior reduction with increasing concentrations of ascorbate leads to partial reduction of cytochrome c1, and a gradual decrease in the magnitude of the oxidation phase of cytochrome b. At an ascorbate concentration sufficient to reduce cytochrome c1 almost completely, the reduction of cytochrome b by either succinate or duroquinol becomes monophasic. Owing to the presence of a trace amount of cytochrome oxidase in the reductase preparation employed, the addition of cytochrome c makes electron flow from substrate to oxygen possible. Under such circumstances, the addition of a limited amount of either succinate or duroquinol leads to a multiphasic reduction and oxidation of cytochrome b. After the initial three phases as described previously, cytochrome b becomes oxidized before cytochrome c1 when the limited amount of added substrate is being used up. However, at the end of the reaction when cytochrome c1 is being rapidly oxidized, cytochrome b becomes again reduced. The above observations support a cyclic scheme of electron flow in which the reduction of cytochrome b proceeds by two different routes and its oxidation controlled by the redox state of a component of the respiratory chain. PMID- 6295472 TI - Identification and quantitation of electron-transport components in human polymorphonuclear neutrophils. AB - Using dithionite difference spectra we have detected cytochrome b in highly purified human neutrophils at a concentration of 0.08 nmol/mg protein. The presence of quinone was identified in lipid extracts at a concentration of approx. 0.06 nmol/mg protein. It was identified as ubiquinone-10 by mass spectrographic analysis. Simultaneous measurements of cytochrome oxidase indicated that these compounds could not be attributed to mitochondrial contamination. These results are compatible with the hypothesis that initiation of the respiratory burst in human neutrophils involves a multicomponent electron transport system. PMID- 6295473 TI - Bioenergetics and protein synthesis. PMID- 6295474 TI - Thiobacillus ferrooxidans. The bioenergetics of an acidophilic chemolithotroph. PMID- 6295475 TI - A spin label study of the lipid boundary layer of mitochondrial NADH-ubiquinone oxidoreductase. AB - Mitochondrial NADH-ubiquinone oxidoreductase (Complex I) is a lipoprotein enzyme containing phosphatidylcholine (PC), phosphatidylethanolamine (PE) and cardiolipin. Enzyme preparations containing endogenous cardiolipin and a range of either soyabean PC or dimyristoylphosphatidylcholine (DMPC) concentrations have been made. Using a spin-labelled fatty acid, two probe environments differing in mobility have been shown to be present. The fatty acid probe has a relative binding constant (or partition coefficient between lipid and protein) of unity. The boundary layer or lipid annulus reported by the probe has a value of approx. 300 lipid molecules per molecule of enzyme FMN in preparations containing soyabean PC, or DMPC above the phase transition temperature of the latter. In soyabean PC-replaced enzyme the apparent size of the boundary layer is independent of temperature between 30 degrees C and 14 degrees C but shows a modest increase to about 400 lipid molecules per molecule of FMN between 14 degrees C and 2 degrees C. Complex I replaced with high concentrations of DMPC gives non-linear Arrhenius plots of NADH-ubiquinone oxidoreductase activity. The results of the ESR experiments show that both boundary layer and bulk lipid must be motionally restricted for this to occur. Thus, the change in activity is probably not caused by an effect exerted directly on the catalytic activity of the enzyme but is more likely due to restriction of free diffusion of ubiquinone to its site of reduction. PMID- 6295476 TI - The effects of lipid fluidity on the rotational diffusion of complex I and complex III in reconstituted NADH-cytochrome c oxidoreductase. AB - NADH-ubiquinone oxidoreductase (Complex I) can be recombined with ubiquinol cytochrome c oxidoreductase (Complex III) to reconstitute NADH-cytochrome c oxidoreductase. Two modes of interaction have been found. In one, the Complexes interact stoichiometrically in one to one molar ratios to give a binary Complex I III unit. In the other, the kinetics of NADH-cytochrome c oxidoreductase are characteristic of 'Q-pool' behaviour seen in intact mitochondria and submitochondrial particles in which the Complexes need not interact directly but can do so via a pool of mobile ubiquinone. Stoichiometric behaviour is found when only boundary layer or annular lipid is present or the lipid is in the gel phase. The lipid is immobile on the ESR time scale and protein rotational diffusion, measured by saturation transfer ESR, is very slow. Q-pool behaviour is found when mobile extra-annular lipid phase is also present. Protein rotational diffusion is rapid and characteristic of a fully disaggregated state. We have also used freeze fracture electron microscopy of reconstituted NADH-cytochrome c oxidoreductase to monitor protein aggregation and lateral phase separation of lipids and proteins under various conditions. We discuss our findings in relation to models for lateral interactions between respiratory chain enzymes. PMID- 6295477 TI - The regulation by cell density of amino acid transport system L in SV40 3T3 cells. AB - The rate of transport of phenylalanine by System L has been measured in SV40 3T3 cells at various cell densities. When the activity of the L system was determined before any cell depletion of intracellular amino acids, a density-dependent increase in transport paralleled the decrease in cell density. This regulation was lost after cell depletion but reappeared after reloading the cells with pertinent substrates of System L. The phenylalanine transport activity modulated by cell density appeared to be related to the internal level of amino acids capable of exchange up to a definite concentration, beyond which transport activity by System L did not parallel a further increase of internal substrate level. Analysis of the relationship between influx and substrate concentration suggested that two saturable components contribute to entry of phenylalanine and leucine in depleted and in reloaded cells: a low-affinity and a high-affinity component. Both kinetic parameters of the high-affinity component appeared to be modulated by the loading treatment, but only V changed markedly. Activation energies for the high-affinity component of the amino acid transport reaction were calculated from an Arrhenius plot in reloaded cells, and were found to be different for low- and high-density cultures. This result is consistent with the interpretation that cell density modulated the rates at which the amino acid carrier complex can move within the cell membrane. PMID- 6295478 TI - Possible existence of ion pairs at the mouths of ion channels. AB - An electrostatic calculation suggests that when an ion is bound near the mouth of a channel penetrating a low-dielectric membrane, a counter ion may form an ion pair with this ion. The tendency towards ion-pair formation is remarkably enhanced at channel mouths by forces (image forces) arising from the charges induced on the boundaries between different dielectrics. The binding constant for the formation of ion-pairs of monovalent ions is estimated under the assumption that local interactions between the counter ion and the channel wall are negligibly small. It is of the order of 1-10 molal-1 or more for the binding of a Cl- (F-) counter ion to an Na+ (Li+) ion if appropriate conditions are fulfilled. The binding constant depends on the position of the binding site, the dimensions and geometries of the channel and channel mouth, and the state of ion loading of the channel, as well as the ionic species. The present results also indicate that when cation (anion) channels have anionic (cationic) groups as integrant parts of their channel walls, interactions between these charged groups and permeant ions are markedly enhanced by the image forces. PMID- 6295479 TI - Probing the pore size of the hemocyanin channel. AB - We have studied single-channel conductance for different monovalent cations and streaming potentials caused by osmotic gradients of non-electrolytes in hemocyanin-treated membranes. We have found that the smaller ion, which cannot pass through the channel, is tetramethylammonium and that acetamide is the smaller non-electrolyte excluded from the pore. From the streaming potentials measured, we calculated that no more than three water molecules can accompany the ion through the channel in a row. From these results we conclude that the hemocyanin channel has in its structure a narrow portion which can be represented as a cylinder 6 A long and 5 A in diameter. PMID- 6295480 TI - A survey of differences between membrane polypeptides of transformed and nontransformed chick embryo fibroblasts. AB - Plasma membrane-associated polypeptides of chick embryo fibroblasts and cells transformed by the Schmidt-Ruppin wild-type strain of Rous sarcoma virus and its temperature-sensitive tsNY68 mutant were compared by two-dimensional gel electrophoresis. Polypeptide and glycoprotein alterations were identified after incubation of cells with [35S]methionine and [3H]mannose and by staining of the gels with 125I-labeled concanavalin A and Coomassie brilliant blue. Polypeptides found to be consistently transformation-sensitive included a group of five polypeptides that were detected only by short-term labeling with methionine, fibronectin, a 180 kDa polypeptide with a pI of 5.6, a mannose-containing glycoprotein of 48 kDA and an unusually high pI of 8.4, and a 19 kDa polypeptide with a pI of approx. 4.5. Several of these polypeptides appear to be particularly interesting for further characterization. PMID- 6295481 TI - Removal of Na+ channels in squid giant axons by perfusion with trypsin. AB - The irreversible effects of the proteolytic enzyme trypsin on ionic and gating currents of voltage-clamped squid axon membranes have been studied. At physiological pH, internal perfusion of the fibre with trypsin was found to be very effective in removing Na+ channels leaving the potassium system almost unaltered. At T = 13 degrees C the rates of channel-cleavage averaged 1/10 min-1 for the Na+ and 1/128 min-1 for the K+ channel, respectively. As estimated by the decrement of peak sodium conductance, the rate of loss of Na+ channels correlates well with the rate of decrease of the total charge associated with the ON component of gating currents, indicating that trypsin probably interacts with an essential proteic portion of the channel whose removal might prevent both the displacement of gating charges and the subsequent opening of the channel. Intracellular pH remarkably influences the action of the enzyme. A plot of the pH dependence of the rate of cleavage of Na+ channels suggests the involvement of a positively charged group (either lysine or arginine) in the substrate region of the trypsin catalytic reaction. PMID- 6295482 TI - Affinity labeling of a nerve growth factor receptor component on rat pheochromocytoma (PC12) cells. AB - Clonal PC12 rat pheochromocytoma cells were sequentially incubated with 125I labeled nerve growth factor and the photoreactive bifunctional agent hydroxysuccinimidyl-p-azidobenzoate. This treatment effected the crosslinking of 125I nerve growth factor to a PC12 cell component that exhibits an apparent Mr = 148 000-158 000, and consists of a single polypeptide chain with internal disulfide bonds. The amount of label associated with this Mr = 148 000-158 000 species was proportional to the degree of occupancy of nerve growth factor receptors by 125I-labeled nerve growth factor. Affinity labeling of this species was inhibited by the presence of 0.2 microM unlabeled nerve growth factor during incubation of PC12 cells with 125I nerve growth factor. In membranes prepared from PC12 cells hydroxysuccinimidyl-p-azidobenzoate effected the crosslinking of 125I-labeled nerve growth factor to an Mr = 120 000-130 000 species but not to the Mr = 148 000-158 000 component observed in intact cells. The kinetics of 125I nerve growth factor affinity labeling of the Mr = 148 000-158 000 species closely paralleled the time-course of 125I nerve growth factor association to two kinetically distinct forms of nerve growth factor receptors in PC12 cells. The data indicate that the Mr = 148 000-158 000 species affinity-labeled by 125I nerve growth factor is the native form of a component associated with kinetically different nerve growth factor receptors in PC12 cells. PMID- 6295483 TI - Characterization of partially purified (Na+ + K+)-ATPase from porcine lens. AB - The partial purification of (Na+ + K+)-ATPase from pig lens has been achieved by treatment with deoxycholate followed by density gradient centrifugation. The specific activity of the final preparation, ranging from 300 to 500 nmol/h per mg protein, is increased approx. 100-fold compared to the homogenate. A parallel increase in rho-nitrophenylphosphatase activity is also observed. Sodium dodecyl sulfate (SDS) gel electrophoresis reveals six major protein bands, one of which is the 93 kDa alpha subunit of (Na+ + K+)-ATPase which can be phosphorylated by reaction with [gamma-32P]ATP. A second band contains a glycoprotein which displays an apparent molecular weight of 51000 and thus appears to be the beta subunit of the enzyme. The enzyme is sensitive to ouabain with the I50 for (Na+ + K+)-ATPase and rho-nitrophenylphosphatase inhibition being 1.2 and 1.3 microM, respectively. Several agents which inhibit (Na+ + K+)-ATPase from other tissues such as oligomycin, Ca2+, vanadate, N-ethylmaleimide, rho chloromercuribenzenesulfonic acid (PCMBS) and 5,5'-dithiobis-(2-nitrobenzoic acid) (DTNB) also inhibit the lens enzyme. Monovalent cations other than K+ are partially effective in activating the (Na+ + K+)-ATPase and rho nitrophenylphosphatase activities. The K+ congeners were relatively more effective in supporting (Na+ + K+)-ATPase compared to rho-nitrophenylphosphatase activity. Other kinetic properties of the lens enzyme are also comparable to those of the enzyme from other tissues. Utilizing the partially purified membrane bound enzyme, discontinuities in Arrhenius plots of (Na+ + K+)-ATPase activity, rho-nitrophenylphosphatase activity and fluorescence polarization of the fluidity probe, 1,6-diphenyl-1,3,5-hexatriene (DPH), are observed near the physiological temperature of lens. The possible significance of these observations for the mechanism of cataract formation are discussed. PMID- 6295484 TI - Microvesiculation and sphingomyelinase activation in chicken erythrocytes treated with ionophore A23187 and Ca2+. AB - Treatment of chicken erythrocytes with ionophore A23187 and Ca2+ leads to the disappearance of the marginal band of microtubules and to a release of the constraints which normally maintain the nucleus in a central position in the cells. The consequent close apposition of the nucleus to the plasma membrane may allow nuclear-plasma membrane fusion to occur and subsequently results in the release of microvesicles from the hybrid surface membrane. The remnant cells are spherical, and have nuclei which appear to be partly exocytosed. Concomitant with these morphological changes, there is a breakdown of 20-30% of the total cell sphingomyelin by an endogenous sphingomyelinase which does not require Ca2+ and which releases phosphorylcholine only into the cell interior. It is suggested that the pool of sphingomyelin which is broken down as a consequence of Ca2+ entry into the cells is present in the nuclear membrane and that it becomes available to the plasma membrane sphingomyelinase as a result of the close apposition of nucleus and plasma membrane induced by Ca2+. PMID- 6295485 TI - Fatty acid acylation of eucaryotic cell membrane proteins. PMID- 6295487 TI - The effect of isoproterenol and hydroxyurea on the presence of ubiquitin and protein A24 in the rat salivary gland. AB - The in vivo administration of hydroxyurea for 12 h counteracts DNA synthesis and cell cycling stimulated by 72 h of isoproterenol treatment in rat salivary gland, as determined by fluorescence-activated flow cytometry. Hydroxyurea has little effect on [3H]leucine incorporation (protein synthesis) of the nuclear proteins soluble in 0.35 M NaCl, when examined by polyacrylamide gel chromatography and autoradiography from electrostatically sorted nuclei of (G0 + G1) and (G2 + M) phases of the in vivo cell cycle. Differential incorporation of [3H]leucine into nuclear proteins was observed during various phases of the cell cycle. Proteins 'X' and 'Z', observed in stained gel chromatographs of the 0.35 M NaCl-soluble nuclear proteins, were identified by biochemical analyses as ubiquitin and protein A24, respectively. Ubiquitin appeared transiently while A24 increased in gel chromatograms concomitant with progressive quiescence of the salivary gland induced by hydroxyurea. PMID- 6295486 TI - Arrangement of proteins in the mitochondrial inner membrane. PMID- 6295488 TI - Two Fe2+ atoms are present in activated prolyl 4-hydroxylase. AB - A linear relationship was found between the activity of prolyl 4-hydroxylase (EC 1.14.11.2) and the amount of Fe2+ bound per mol enzyme. At maximal activity (2.1 mumol X min-1 per mg protein) the enzyme contains 2.1 mol Fe2+ specifically bound per mol enzyme tetramer, indicating two Fe2+-binding sites on prolyl 4 hydroxylase. The half-maximal concentration of added Fe2+ for enzyme activity depends on the nature of sulphydryl compounds present in the reaction medium. PMID- 6295489 TI - Characterization of cyclic 3':5'-amp-dependent protein kinase in sarcoplasmic reticulum and cytosol of canine myocardium. AB - Canine cardiac sarcoplasmic reticulum vesicles contain intrinsic cAMP-dependent and Ca2+ -calmodulin-dependent protein kinase (EC 2.7.1.37) activities and a common substrate, phospholamban, for these enzymes. Cyclic AMP-dependent protein kinase associated with sarcoplasmic reticulum membranes was solubilized with Triton X-100. Solubilization of the sarcoplasmic reticulum protein kinase did not affect its dependency on cAMP or its substrate specificity. The solubilized cAMP dependent protein kinase was purified by DEAE-cellulose chromatography and was characterized as a type II enzyme on the basis of its elution at high ionic strength. DEAE-purified cAMP-dependent protein kinase exhibited no Ca2+ calmodulin-dependent protein kinase activity. Cytosol from canine cardiac muscle cells, chromatographed on DEAE-cellulose under conditions identical to those used with sarcoplasmic reticulum, exhibited the presence of both type I and type II cAMP-dependent protein kinase isozymes. The properties of the DEAE-cellulose purified type II protein kinases from sarcoplasmic reticulum and cytosol were similar. We conclude that cardiac sarcoplasmic reticulum contains primarily type II cAMP-dependent protein kinase and this is probably the enzyme which phosphorylates sarcoplasmic reticulum in vivo and regulates Ca2+ transport. PMID- 6295490 TI - Production of superoxide anion by N,N-bis(2-hydroxyethyl) iminotris(hydroxymethyl)methane buffer during oxidation of oxyhemoglobin by nitrite and effect of inositol hexaphosphate on the oxidation. AB - Oxidation of oxyhemoglobin by nitrite is characterized by the presence of a lag phase followed by an autocatalysis. As reported previously (Kosaka, H., Imaizumi, K. and Tyuma, I. (1982) Biochim. Biophys. Acta 702, 237-241), in phosphate buffer nitrite produced an ESR signal at g 2.005 (hereafter referred to as the g 2 radical). The g 2 radical produced NO.2 from NO-2, then NO.2 oxidized oxyhemoglobin. Superoxide dismutase did not modify the oxidation. On the other hand in N,N-bis(2-hydroxyethyl)iminotris(hydroxymethyl)methane (bistris) buffer, superoxide dismutase markedly elongated the lag phase and accelerated the autocatalysis, indicating O-2 production. Bistris scavenged the g 2 radical. O-2 was generated by the reduction of O2 by a radical derived from bistris. Inositol hexaphosphate inhibited the oxidation by decreasing H2O2 production from oxyhemoglobin. PMID- 6295491 TI - The peroxidation of thiocyanate catalysed by myeloperoxidase and lactoperoxidase. AB - Peroxidation of SCN- to OSCN-, catalysed by myeloperoxidase and lactoperoxidase, was studied. The rate of this reaction showed sharp optima between pH 5 and 7.5, the position of which is determined by the concentrations of both SCN- and H2O2. At low pH values, both SCN- and H+ inhibited myeloperoxidase and lactoperoxidase competitively with respect to H2O2. The inhibition constants of SCN- for myeloperoxidase and lactoperoxidase (2 and 6 mM, respectively) are independent of pH. For these enzymes a Ki for H+ of 1 microM was found that corresponded to an ionisable group on the enzymes (pKa = 6) which controls the enzymic activity. A kinetic expression is proposed that explains most of the data. The physiological consequences of the corresponding mechanism are discussed. PMID- 6295492 TI - Deoxyguanosine kinase from human placenta. AB - Deoxyguanosine kinase (ATP:deoxyguanosine 5'-phosphotransferase) has been purified up to a specific activity of 10.3 nmol/min per mg protein from human placenta. The enzyme appears to have a molecular weight of 58 000 from the results of Sephadex G-75 gel filtration. The enzyme catalyzed phosphorylation of deoxyguanosine and deoxyadenosine, but deoxycytidine was not phosphorylated. An apparent Km value for deoxyguanosine was 2.5 micro M. When ATP was used as a phosphate donor, the pH optimum was at pH 6.0, but the optimum was shifted to pH 6.8 by the addition of dTTP. At physiological pH, the activity was stimulated 3-4 fold by dTTP. dTTP was also an effective phosphate donor, but using dTTP as a phosphate donor, a broad pH optimum of 7.0 was observed. Two Km values of 0.13 and 2.2 mM were obtained for both MgATP2- and MgdTTP2-. The activity was strongly inhibited by dGTP and dGDP; 50% inhibition by 1.0 micro M dGTP and 2.1 micro M dGDP, respectively. The enzyme required the presence o Mg2+ or Mn2+. PMID- 6295493 TI - A comparison of the heme electronic states in equilibrium and nonequilibrium protein conformations of high-spin ferrous hemoproteins. Low temperature magnetic circular dichroism studies. AB - The visible and near infrared magnetic circular dichroism (MCD) spectra of equilibrium high-spin ferrous derivatives of myoglobin, hemoglobin, horseradish peroxidase and mitochondrial cytochrome c oxidase at 15 K are compared with those of the corresponding proteins in nonequilibrium conformations produced by low temperature photodissociation of CO-complexes of these proteins as well as of O2 complexes of myoglobin and hemoglobin. Over all the spectral region (450-800 nm) the intensities of MCD bands of hemoproteins studied in equilibrium conformation are shown to be strongly temperature-dependent, including a negative band at ca. 630 nm and positive bands at ca. 690 nm and at ca. 760 nm. In contrast to the absorption spectra, the low-temperature MCD spectra of high-spin ferrous hemoproteins differ significantly, reflecting the peculiarities in the heme iron coordination sphere which are created by a protein conformation. The MCD spectra reveal clearly the structural changes in the heme environment which occur on ligand binding. On the basis of assignment of d leads to d and charge-transfer transitions in the near infrared region the correlation is suggested between the wavelength position of the MCD band at approx. 690 nm and the value of iron out of-plane displacement as well as between the location of the band at approx. 760 nm and the Fe-N epsilon (proximal histidine) bond strength (length) in equilibrium and nonequilibrium conformations of the hemoproteins studied. The high sensitivity of low-temperature MCD spectra to geometry at heme iron is discussed. PMID- 6295494 TI - Distinctive selectivity for docosatetraenoic acid incorporation by Ehrlich ascites tumor cells. AB - Three polyunsaturated fatty acids were incorporated into the lipids of Ehrlich ascites cells to examine the selectivity of the synthetic enzymes for chain length and the number of double bonds. Arachidonic acid (20:4n-6) and timnodonic acid (20:5n-6) showed approximately similar patterns of incorporation into neutral acylglycerols, choline glycerophospholipids and ethanolamine glycerophospholipids. Further division of the lipids into diacyl and alkylacyl categories established that 20:4n-6 was incorporated at relatively lower amounts in the triacylglycerols and in the alkylacyl and alkenylacyl forms of the glycerophospholipids. The pattern of incorporation of radioactive 20:5n-3 into the phospholipids was similar to that for 20:4n-6, but there was no evidence for a selectively restrained incorporation into the triacylglycerols. The 22-carbon acid (22:4n-6) differed greatly from its shorter homolog (20:4n-6) in being incorporated in much greater amounts into triacylglycerols than into phospholipids. It did not appear to be present at an appreciably higher relative amount in either the alkyl diacylglycerols or in the ether-containing choline glycerophospholipids. Nevertheless, it occurred with exceptionally high specific activities in the alkylacyl and alkenylacyl forms of ethanolamine glycerophospholipids in a manner that suggests marked discrimination by the cytidine-mediated ethanolamine phosphotransferase. This distinctive selectivity provides evidence for special role for a 22-carbon acid in lipid metabolism. PMID- 6295495 TI - Glycosylation of LDL decreases its ability to interact with high-affinity receptors of human fibroblasts in vitro and decreases its clearance from rabbit plasma in vivo. AB - Incubation of human LDL in vitro at 37 degrees C for 48 h with [14C]glucose at concentrations from 5 to 200 mM resulted in a glycosylated LDL, containing 0.4-20 mol of glucose incorporated per apolipoprotein B of 250 000 daltons. The extent of glucose incorporated was proportional to the time of incubation and concentration of glucose. Glycosylation of LDL abolished its uptake and degradation by the high-affinity process for LDL in normal human skin fibroblasts. 125I-labeled glycosylated LDL was bound, internalized and degraded by the fibroblasts via a nonspecific low-affinity process. The 125I-labeled glycosylated LDL and 125I-labeled LDL were taken up and degraded at similar rates in a non-saturable, low-affinity process by peritoneal macrophages isolated from mice. When 125I-labeled glycosylated LDL or 125I-labeled LDL were injected into rabbits, the glycosylated LDL had a delayed plasma clearance in comparison to the LDL. The mean fractional catabolic rates were 0.67 day-1 and 1.70 day-1 for 125I labeled glycosylated LDL and 125I-labeled LDL, respectively. The uptake and degradation of 125I-labeled LDL by human skin fibroblasts was decreased as the concentration of free carbohydrate, glucose, sucrose or sorbitol, in the medium was increased from 10 mM to 1 M. It is speculated that pathologic levels of plasma glucose in vivo could result in a decrease in LDL uptake as a result of glycosylation of LDL. A decrease in uptake of native or modified LDL in vivo could contribute to hypercholesterolemia and its pathophysiology. PMID- 6295496 TI - Binding properties of high-density lipoprotein subfractions and low-density lipoproteins to rabbit hepatocytes. AB - Primary cultures of rabbit hepatocytes which were preincubated for 20 h in a medium containing lipoprotein-deficient serum subsequently bound, internalized and degraded 125I-labeled high-density lipoproteins2 (HDL2). The rate of degradation of HDL2 was constant in incubations from 3 to 25 h. As the concentration of HDL2 in the incubation medium was increased, binding reached saturation. At 37 degrees C, half-maximal binding (Km) was achieved at a concentration of 7.3 micrograms of HDL2 protein/ml (4.06 X 10(-8)M) and the maximum amount bound was 476 ng of HDL2 protein/mg of cell protein. At 4 degrees C, HDL2 had a Km of 18.6 micrograms protein/ml (1.03 X 10(-7)M). Unlabeled low density lipoproteins (LDL) inhibited only at low concentrations of 125I-labeled HDL2. Quantification of 125I-labeled HDL2 binding to a specific receptor (based on incubation of cells at 4 degrees C with and without a 50-fold excess of unlabeled HDL) yielded a dissociation constant of 1.45 X 10(-7)M. Excess HDL2 inhibited the binding of both 125I-labeled HDL2 and 125I-labeled HDL3, but excess HDL3 did not affect the binding of 125I-labeled HDL3. Preincubation of hepatocytes in the presence of HDL resulted in only a 40% reduction in specific HDL2 receptors, whereas preincubation with LDL largely suppressed LDL receptors. HDL2 and LDL from control and hypercholesterolemic rabbits inhibited the degradation of 125I-labeled HDL2, but HDL3 did not. Treatment of HDL2 and LDL with cyclohexanedione eliminated their capacity to inhibit 125I-labeled HDL2 degradation, suggesting that apolipoprotein E plays a critical role in triggering the degradative process. The effect of incubation with HDL on subsequent 125I labeled LDL binding was time-dependent: a 20 h preincubation with HDL reduced the amount of 125I-labeled LDL binding by 40%; there was a similar effect on LDL bound in 6 h but not on LDL bound in 3 h. The binding of 125I-labeled LDL to isolated liver cellular membranes demonstrated saturation kinetics at 4 degrees C and was inhibited by EDTA or excess LDL. The binding of 125I-labeled HDL2 was much lower than that of 125I-labeled LDL and was less inhibited by unlabeled lipoproteins. The binding of 125I-labeled HDL3 was not inhibited by any unlabeled lipoproteins. EDTA did not affect the binding of either HDL2 or HDL3 to isolated liver membranes. Hepatocytes incubated with [2-14C]acetate in the absence of lipoproteins incorporated more label into cellular cholesterol, nonsaponifiable lipids and total cellular lipid than hepatocytes incubated with [2-14C]acetate in the presence of any lipoprotein fraction. However, the level of 14C-labeled lipids released into the medium was higher in the presence of medium lipoproteins, indicating that the effect of those lipoproteins was on the rate of release of cellular lipids rather than on the rate of synthesis. PMID- 6295498 TI - Control of phosphatidylinositol turnover in adrenal glomerulosa cells. AB - The purpose of the present experiments was to compare the effects on phosphatidylinositol metabolism of agents stimulating aldosterone secretion. Glomerulosa cells, isolated from rat adrenals, were incubated in the presence of one of the following stimuli: angiotensin II, elevated potassium concentration, corticotropin, dibutyryl cyclic AMP and prostaglandin E2. Of all these substances, only angiotensin II stimulated the incorporation of [32P]phosphate into phosphatidylinositol. The effect was already detected 2.5 min and was still maintained 60 min after the onset of stimulation. A slight enhancement of the incorporation into other phospholipids was observed in the first minutes of stimulation. Cycloheximide abolished the effect of angiotensin II on aldosterone production, but not on phosphatidylinositol synthesis. In cells prelabelled with [32P]phosphate, radioactivity in phosphatidylinositol relative to that in other phospholipids decreased in response to angiotensin II within 5 min. This indicates that angiotensin II induces a specific breakdown of phosphatidylinositol. Corticotropin failed to enhance the incorporation of [32P]phosphate into phosphatidylinositol and other phospholipids in isolated fasciculate-reticularis cells. The results suggests that although both angiotensin II and potassium are presumed to act through changes in calcium metabolism, angiotensin alone generates the calcium signal by increased phosphatidylinositol turnover. PMID- 6295497 TI - Mechanism of phospholipid peroxidation induced by ferric ion-ADP-adriamycin-co ordination complex. AB - A system, which contains NADPH, purified cytochrome P-450 reductase and adriamycin, produces H2O2, O-2 and adriamycin semiquinone radical with O2 consumption and NADPH oxidation. This system, however, does not promote a peroxidation cleavage of unsaturated phospholipid. On the other hand, ferric ion ADP-adriamycin-co-ordination complex, which may convert to a perferryl ion-co ordination complex by an intramolecular electron transfer mechanism in air, acts as a powerful initiator for lipid peroxidation. A similar perferryl ion-co ordination complex could also be produced from ferric ion-ADP-adriamycin-co ordination complex after reducing it by NADPH-dependent cytochrome P-450 reductase in air. PMID- 6295499 TI - Interactions between prostaglandin precursors during their oxygenation by human platelets. AB - The oxygenation through the prostaglandin synthase complex and the lipoxygenase pathways of the three prostaglandin precursors was investigated in human platelets. These precursors (dihomogammalinolenic (8,11,14-eicosatrienoic; 20:3 (8,11,14)), arachidonic (5,8,11,14-eicosatetraenoic; 20:4 (5,8,11,14)) and 5,8,11,14,17-eicosapentaenoic (20:5 (5,8,11,14,17)) acids) were used alone or simultaneously. We have found that 20:4 (5,8,11,14) increases the oxygenation of 20:3 (8,11,14) by the prostaglandin synthase complex while 20:5 (5,8,11,14,17) decreases the oxygenation of 20:4 (5,8,11,14) by the same enzyme complex. On the other hand, the utilization of 20:5 (5,8,11,14,17) by the prostaglandin synthase complex and the lipoxygenase was markedly enhanced in the presence of 20:3 (8,11,14), 20:4 (5,8,11,14) or both. Besides, the increased concentrations of 20:5 (5,8,11,14,17) failed to enhance its oxygenation to such an extent while the addition of 20:3 (8,11,14) or 20:4 (5,8,11,14) allows the marked potentiation of the 20:5 (5,8,11,14,17) oxygenation at any concentration. This indicates that, to be efficient, the utilization of 20:5 (5,8,11,14,17) by platelet oxygenases needs the presence of 20:3 (8,11,14), 20:4 (5,8,11,14) or their derivatives. In addition, using small concentrations of each prostaglandin precursor close to concentrations presumably released from platelet phospholipids during aggregation, all show the same tendencies. We conclude that the interactions we have observed between prostaglandin precursors during their oxygenation by human platelets could be of primary importance to explain the modifications of platelet reactivity reported after dietary manipulations. PMID- 6295500 TI - Receptor-mediated catabolism and tissue uptake of human low density lipoprotein in the cholesterol-fed, atherosclerotic rabbit. AB - The LDL receptor pathway, which was delineated in cultured cells, is now known to operate in vivo. In this study we have measured the plasma clearances and tissue uptakes of native and chemically modified (1,2-cyclohexanedione-treated or reductively methylated) LDL in rabbits in order to determine the response of the pathway to a high-cholesterol diet. 1 week on the diet increased circulating LDL and suppressed its receptor-mediated plasma clearance and uptake into all tissues. The fractional catabolic rate of the lipoprotein via the receptor independent route also fell. Continuation of the feeding program for 12 weeks accentuated these changes and virtually eliminated receptor uptake into all tissues so that the plasma decay curves of native and cyclohexanedione-treated LDL were superimposable. Lipoprotein assimilation by the aorta, however, did not follow this general trend. This tissue, after 12 weeks, was variably infiltrated by atheromatous deposits and the appearance of these lesions was associated with a substantial increase in the relative uptakes of both native and chemically modified (cyclohexanedione-treated and reductively methylated) LDL. We concluded (a) that expansion of tissue cholesterol pools virtually abolishes LDL receptor activity in rabbits; and (b) that LDL assimilation (both apparently receptor mediated and receptor-independent) paradoxically increases at sites where the aorta is affected by atheromatous lesions. PMID- 6295501 TI - Formation of alkylacyl- and diacylglycerophosphocholines via diradylglycerol cholinephosphotransferase in rat liver. AB - We found that diacylglycerol and alkylacylglycerol choline phosphotransferase activities exhibited similar pH optima, thermolabilities and inhibitions by Mn2+, and dithiothreitol. The Vmax of diacylglycerol cholinephosphotransferase was higher (approx. 1-2-fold) than the Vmax of alkylacylglycerol cholinephosphotransferase. The Km value for diacylglycerol was somewhat greater than for the alkylacylglycerol, but no differences were found for the Km of CDPcholine with either type of diradylglycerol as the other substrate. Endogenous levels of diacylglycerols in microsomes were 24.3 nmol/mg protein, whereas no detectable amount of alkylacylglycerols was observed. Results from this study indicate that a similar or identical enzyme catalyzes the formation of 1,2-diacyl and 1-alkyl-2-acyl-sn-glycero-3-phosphocholine and that the availability of diradylglycerols and the turnover rate of ether-linked lipids would appear to be important factors in controlling the level (under 1%) of 1-alkyl-2-acyl-sn glycero-3-phosphocholine in rat liver. PMID- 6295502 TI - Properties of diacylglycerol kinase in adult and fetal rat lung. AB - Diacylglycerol kinase activity is found in both adult and fetal lung. Approximately 27 and 52% of the total activity is found in microsomes and cytosol, respectively. The activity is maximal at pH 7.4. The apparent Km for ATP is 0.11 mM and 0.21 mM for cytosol and microsomes, respectively. The apparent Km for dioleoylglycerol is 0.05 mM for cytosol and 0.14 for microsomes. Maximal activity in cytosol and microsomes is obtained with 2.0 mM dexoycholate. Other detergents cannot substitute for deoxycholate. Phosphatidylglycerol stimulates activity in the absence and in the presence of deoxycholate. Phosphatidylserine also stimulates activity, whereas phosphatidylethanolamine was inactive and phosphatidylcholine inhibited the reaction. Linoleic acid produced inhibition. The general properties of the enzyme were similar for fetal and adult lung. Diacylglycerol kinase from microsomes and cytosol fraction from both fetal and adult lung was most active with dioleoylglycerol and diacylglycerol from egg phosphatidylcholine. Significantly lower activity was obtained with dipalmitoylglycerol. Phosphatidylglycerol did not alter the relative substrate preferences. The activity in microsomes increased with development from 19 days gestation to a maximal activity at 21 days gestation. Maximal activity was about 2-fold higher than the adult. The activity dropped rapidly reaching adult values prior to birth (22 days gestation). The activity in cytosol fractions increased gradually from 19 days gestation, reaching adult values by 22 days gestation. PMID- 6295503 TI - Cyclic AMP-binding proteins in human blood platelets detected by photoaffinity labelling. AB - Cyclic AMP inhibits platelet aggregation induced by physiological agents. 8 Azido [32P]cyclic AMP (N3 cyclic AMP) has been utilized as a photoaffinity probe to define the cyclic AMP-binding proteins present in unperturbed human platelets and their subcellular fractions. Specificity of cyclic AMP binding was determined by contrasting binding in the presence and absence of excess unlabelled cyclic AMP, cyclic GMP and 5'-AMP. Binding was unaffected by 5'-AMP and obliterated by cyclic AMP. Four major species of binding proteins, 49 000, 42 000, 39 000, 37 000, were obtained in all platelet fractions (crude homeogenate, cytosol, membranes and granules). Two-dimensional gel electrophoresis of platelet cytosol resolved the major molecular weight species into 15 specific cyclic AMP binding proteins of four molecular weight classes differing by charge density. These studies suggest that platelets contain an array of specific cyclic AMP-binding proteins which may function in hemostatic regulation. PMID- 6295504 TI - Isolation and characterization of monoclonal antibodies to (Na+ + K+)-ATPase. AB - Four stable hybridoma cell lines secreting antibodies specific to the membrane (Na+ + K+)-dependent ATPase isolated from lamb kidney medulla have been produced by fusing mouse myeloma cells with spleen cells from immunized mice. These cell lines produce IgG gamma 1 heavy chain and kappa light chain antibodies which are directed against the catalytic or alpha-subunit of the (Na+ + K+)-ATPase enzyme. Binding studies, using antibodies that were produced by growing hybridomas in vivo and purified by affinity column chromatography, suggest a somewhat higher affinity of these antibodies for the isolated alpha-subunit than for the 'native' holoenzyme. In addition, these monoclonal antibodies show no reactivity with either the glycoprotein (beta) subunit of the lamb enzyme nor the (Na+ + K+) ATPase from rat kidney, an ouabain-insensitive organ. Cotitration binding experiments have shown that the antibodies from two cell lines originally isolated independently from the same culture plate well population of fused cells bind to the same determinant site and are probably the same antibody. Cotitration and competition binding studies with two other antibodies have revealed two additional distinct antibody binding sites which appear to have little overlap with the first site. One of the three different antibodies isolated caused a partial inhibition of the (Na+ + K+)-ATPase activity. This antibody appears to be directed against a specific functionally important site of the alpha-subunit and is a competitive inhibitor of ATP binding. Under optimum conditions of ATPase activity, this inhibitory effect is not altered by the presence of the other two antibodies. PMID- 6295505 TI - Production of plasminogen activator by synchronized normal and Rous sarcoma virus transformed chicken fibroblasts in culture. AB - We studied intracellular activity of the plasminogen activator within the cell cycle of chemically synchronized normal and RSV-transformed chick fibroblasts in culture. Consideration has also been given to the relationship between the plasminogen activator activity and cycles of DNA synthesis or mitosis in cycling fibroblasts after viral infection. The plasminogen activator activity of the cell lysates was assayed on [125I]fibrin-coated Petri dishes and was expressed as the radioactivity released from the plates. Normal fibroblasts produced detectable levels of plasminogen activator in the S-phase and late G2-phase or mitosis of the cell cycle. In contrast, RSV-transformed cells produced high levels of this activator throughout the entire cell cycle although this activity fluctuated and reached a maximum in the G2-M periods. We also found that the level of plasminogen activator activity in the transformed fibroblasts is influenced by the cycles of DNA synthesis and that cell division is required for the appearance of plasminogen activator activity in the 'de novo' virus-infected cultures. PMID- 6295506 TI - The regulatory role of spermine and fatty acid in the interaction of AMP deaminase with phosphofructokinase. AB - The role of fatty acid and polyamine in the interaction of AMP deaminase (EC 3.5.4.6)-ammonium system with glycolysis was investigated using permeabilized yeast cells. (1) The addition of fatty acid inhibited the activity of AMP deaminase in situ, resulting in a decrease in the total adenylate pool depletion, and in the recovery of the adenylate energy charge. (2) The addition of fatty acid resulted in an indirect decrease in the activity of phosphofructokinase (EC 2.7.1.11) through a reduced level of ammonium ion; fatty acid itself did not inhibit phosphofructokinase activity in the presence of excess ammonium ion. (3) Spermine protected AMP deaminase from inhibition by fatty acid: the increased ammonium level enhanced phosphofructokinase activity, glycolytic flux and the recovery of the energy charge. In contrast, alkali metals, which are also activators of AMP deaminase had little effect on the inhibition of the enzyme. The inhibition of glycolysis by fatty acid and its reversal by polyamine can be accounted for by the changes in ammonium ion through the action of AMP deaminase ammonium system, and the physiological relevance is discussed. PMID- 6295508 TI - Effect of microtubule-disrupting agents on superoxide production in human polymorphonuclear leukocytes. AB - We explored the effects of compounds known or proposed to affect microtubule functions on superoxide (O2-) production in human polymorphonuclear leukocytes stimulated by N-formyl-methionyl-phenylalanine (f-Met-Phe), calcium ionophore A23187 and phorbol myristate acetate. F-Met-Phe-induced O2- production was markedly potentiated not only by microtubule-disrupting agents, including colchicine, vincristine, vinblastine, nocodazole, podophyllotoxin and griseofulvin, but also deuterium oxide (2H2O), which is proposed to stabilize microtubules, and not affected by lumicolchicine. Ionophore A23187-induced O2- production was not influenced by colchicine, and markedly enhanced by 2H2O, whereas phorbol myristate acetate-induced O2- production was not influenced by colchicine, and slightly inhibited by 2H2O. 2H2O did not counteract the effects of colchicine and vice versa. Dibutyryl cyclic AMP and prostaglandin E1 inhibited O2- production stimulated by f-Met-Phe and ionophore A23187, whereas phorbol myristate acetate-induced O2- production was strongly resistant to the inhibitory effect of these agents. The enhancing effect of colchicine and 2H2O on f-Met-Phe induced O2- production was abolished by dibutyryl cyclic AMP. Colchicine promoted concanavalin A cap formation, and 2H2O produced concanavalin A patch formation, whereas dibutyryl cyclic AMP did not affect the distribution of concanavalin A receptors. In addition, 2H2O and dibutyryl cyclic AMP did not interfere with the colchicine-induced concanavalin A cap formation. These findings suggest that f Met-Phe, ionophore A23187 and phorbol myristate acetate may activate the oxidative metabolism of human polymorphonuclear leukocytes through different mechanisms, and that microtubule-disrupting agents, 2H2O and cyclic AMP agonists may affect the different steps of the activating system of NAD(P)H oxidase. PMID- 6295507 TI - Biological activities and spectroscopic properties of chromophoric and fluorescent analogs of adenine nucleoside and nucleotides, 2',3'-O-(2,4,6 trinitrocyclohexadienylidene) adenosine derivatives. AB - The ribose-modified chromophoric and fluorescent analog of ATP 2',3'-O-(2,4,6 trinitrocyclohexadienylidene) adenosine 5'-triphosphate (TNP-ATP) has been synthesized previously (Hiratsuka, T., and Uchida, K. (1973) Biochim. Biophys. Acta 320, 635-647 and Hiratsuka, T. (1976) Biochim. Biophys. Acta 453, 293-297). In the present study, four TNP-derivatives of ATP, ADP, AMP and adenosine were synthesized and compared for several chemical, spectral and enzymatic properties. Their visible absorption and fluorescent properties were found to be quite similar. Visible absorption and fluorescence spectra of TNP-derivatives were sensitive to solvent polarity. TNP-adenosine and TNP-AMP showed considerable substrate activities with adenosine deaminase and alkaline phosphatase, respectively. TNP-ATP proved to be an excellent substitute for ATP in adenylate kinase and myosin ATPase systems. The results indicate that these analogs are useful as chromophoric and fluorescent probes for hydrophobic regions in adenine nucleoside and nucleotide requiring enzymes. PMID- 6295509 TI - Synthesis and composition of vitamin D-3 metabolites in Solanum malacoxylon. AB - The synthesis of vitamin D-3 hydroxylated metabolites in Solanum malacoxylon was investigated. When crude leaf homogenates and subcellular fractions were incubated with [3H]vitamin D-3 and [3H]25-hydroxy-vitamin D-3 under conditions described for animal vitamin D-3-25-hydroxylase and 25-hydroxy-vitamin D-3-1 alpha-hydroxylase, respectively, labelled metabolites identified on the basis of their chromatographic properties as 25-hydroxy-vitamin D-3 and 1,25-dihydroxy vitamin D-3 were formed. Other unidentified product metabolites were also detected. Vitamin D-3-25-hydroxylase activity was localized in microsomes and 25 hydroxy-vitamin D-3-1 alpha-hydroxylase in mitochondria and microsomes. Chromatography of sterols isolated from leaf extracts preincubated with beta glucosidase on Sephadex LH-20 columns permitted the isolation of three biologically active fractions with elution properties similar to vitamin D-3, 25 hydroxy-vitamin D-3 and 1,25-dihydroxy-vitamin D-3, respectively. Ultraviolet spectra characteristic of vitamin D-3 and its metabolites were obtained after purification of the fractions by TLC. Co-chromatography of individual fractions with authentic metabolites on TLC provided further evidence that the plant contains vitamin D-3, 25-hydroxy-vitamin D-3 and 1,25-dihydroxy-vitamin D-3 as glucoside derivatives. These results suggest that a similar pathway of vitamin D 3 hydroxylation as in animals may be operative in S. malacoxylon. PMID- 6295510 TI - Generation of hydrogen peroxide during the oxidation of L-phenylalanine by Proteus mirabilis isolated membranes. AB - In the process of L-phenylalanine oxidation by Proteus mirabilis cytoplasmic membrane, hydrogen peroxide was produced at a rate corresponding to 1-3 per cent of the total electron flow (30-110 nmoles min-1mg-1). Peroxide was estimated using a fluorimetric assay with horseradish peroxidase, or by anodic oxidation on a platinum electrode. When using the former method, superoxide dismutase decreased the apparent yield of peroxide, a fact suggesting that H2O2 was in part the dismutation product of superoxide radicals. However the superoxide dismutase effect could be an artefact due to the generation of some superoxide during the peroxidatic reaction in the assay. Adrenaline was the reagent used for the detection of superoxide. There was no significant emergence of superoxide as the result of phenylalanine oxidation by the membrane (specific activity lower than 1 2 nmoles min-1mg-1). Thus it seemed that superoxide was not an intermediate for the bulk of H2O2 formed in this system. According to the results, peroxide was probably formed at a stage of electron transport earlier than the cytochrome level. The membrane phenylalanine dehydrogenase could be a site where peroxide was evolved in these experiments. PMID- 6295511 TI - Induction of drug metabolizing enzymes in the liver of diabetic mice. AB - The effects of two classical inducers, phenobarbital and 3-methylcholanthrene, have been tested on some liver microsomal drug-metabolizing enzymes (monooxygenases and phase II enzymes) and on benzo(a)pyrene metabolism in genetically (ob/ob) and chemically (streptozotocin) diabetic mice. 1) In ob/ob mice, the basal activities and the inducibility of phase I and phase II enzymes, as well as the electrophoretic pattern of microsomal proteins, were not notably different from those of similarly treated lean mice. 2) A possibly common form of cytochrome P 450 present both in microsomes from steptozotocin-diabetic non induced mice and in those from phenobarbital-treated non-diabetic mice could explain the increased "phenobarbital-like" enzyme activities in chemically diabetic animals. 3) The increase of monooxygenase activities produced by streptozotocin treatment is partially depressed by 3-methylcholanthrene, probably as a result of the dilution of "phenobarbital-like" cytochrome P 450 forms by 3 methylcholanthrene-induced cytochrome P 448. 4) The increased formation of the most carcinogenic metabolites of benzo(a)pyrene, and the slight decrease of phase II conjugation enzyme activities, may add their deleterious effects in 3 methylcholanthrene-induced streptozotocin-diabetic animals. PMID- 6295512 TI - Murine retrovirus genome directs the synthesis of gag protein precursor early after infection. AB - Incoming type C retroviral genomic 35S RNA is present in polysomes of undifferentiated and differentiated murine teratocarcinoma cell lines at 4 hours after infection. At the same time a 65,000 daltons viral specific protein is produced by the infected cells. These data present evidence that incoming viral RNA serves as messenger for the synthesis of gag protein precursor Pr65 early in the infectious cycle of ecotropic murine retrovirus. PMID- 6295513 TI - [Interaction of N1-, N6- and C8-substituted derivatives of adenosine-5' triphosphate with the catalytic subunit of cAMP-dependent protein kinase from rabbit skeletal muscles]. AB - In order to investigate the structure of the active site of the cAMP-dependent protein kinase catalytic subunit a synthesis of several previously unknown adenosine-5'-triphosphate (ATP) derivatives containing substituents of various nature at N(1), N(C6) and C(8) positions of the purine base was carried out. The interaction of these derivatives with a homogeneous preparation of the catalytic subunit of rabbit skeletal muscle cAMP-dependent protein kinase was investigated. All the nucleotide analogs were found to inhibit the enzyme activity; the inhibition was competitive with respect to ATP. It was assumed that the adenine moiety of the ATP molecule is bound to the active site of protein kinase by the hydrophobic interaction with the aromatic amino acid residues and by formation of the hydrogen bond between the exo-NH2-group of the substrate and a corresponding group of the enzyme. The "correct" binding of ATP to the enzyme active center is defined by the anti-conformation of the nucleotide. PMID- 6295514 TI - [Two molecular forms of pea ferredoxin in the electron transport chain of chloroplasts]. AB - The effects of two molecular forms of water-soluble ferredoxin (Fd I and Fd II) on the kinetics of electron transport in bean chloroplasts (class B) were studied. The light-induced redox transitions of the photosystem I reaction center P700 were measured by the intensity of the EPR signal I produced by P700+. Both forms of ferredoxin, Fd I and Fd II, when added to the chloroplasts in catalytic amounts, stimulate the light-induced electron transfer from P700 to NADP+. Nevertheless, Fd I is a better mediator of the back reactions from NADPH to P700+. This electron transfer pathway is sensitive to the cyclic electron transport inhibitor, antimycin A, and to DCMU inhibitor of electron transport between photosystem II and plastoquinone. It may be concluded that the two molecular forms of ferredoxin, Fd I and Fd II, differ in their ability to catalyze cyclic electron transport in photosystem I. The role of Fd I and Fd II in regulation of electron transport at the acceptor site of photosystem I is discussed. PMID- 6295515 TI - [Effect of dehydrocorrine-cobalt complex on mitochondria]. AB - The effects of the decamethyloctadehydrocorrine-cobalt complex (Co-C) on respiration and the ATP-synthetase activity of rat liver mitochondria were investigated. The Co-C complex was found to be an effective shunt of the respiratory chain. It accepts electrons from ubiquinone and donates them directly to O2. The Co-C complex inhibits the ATPase and ATP-synthetase activities of mitochondria. PMID- 6295516 TI - [Structural reconstruction of chemo-sensitive biomembranes during the action of low molecular weight compounds using spin resonance]. AB - Using the spin probe technique, the changes in the supramolecular structure of the central nervous system synaptic membranes and of olfactory hair membranes of Rana temporaria induced by low molecular weight organic substances of different chemical nature, were investigated. It was found that the membrane structures under study differ considerably in their sensitivity to chemical stimulation and in temporal kinetics of structural transitions. A correlation between physiological parameters of olfactory perception of the compounds used and the parameters of structural transitions in olfactory hair membranes was established. The interrelationship between the experimental data and chemoreception of odorants and mediators as well as the applicability of these preparations for membrane screening of biologically active substances are discussed. PMID- 6295517 TI - [Isolation, some properties and heterogeneous nature of DNA-topoisomerase (relaxing enzyme) from Ehrlich ascites carcinoma cells]. AB - DNA-topoisomerase catalyzing the conversion of a superhelical circular covalently closed DNA molecule into a super-helix free circular molecule, was isolated from mouse Ehrlich ascites carcinoma cells and purified 209-fold. The optimal conditions for the action and stability of the enzyme were elaborated. Using polyacrylamide gel electrophoresis under non-denaturating conditions as well as in the presence of Na-DS the heterogeneity of purified DNA-topoisomerase was established. This heterogeneity implies the presence of three active forms of the enzyme with Mr of 97 000, 81 000 and 69 000, respectively. Using one-dimensional fingerprint method and limited proteolysis with Staphylococcus aureus protease, it was demonstrated that the low molecular weight enzyme forms are products of limited proteolysis of the highest molecular weight form of DNA-topoisomerase. PMID- 6295518 TI - Clinical studies of the endogenous opioid system. AB - The role of the endogenous opioid system in humans was studied using three clinical research strategies. High doses of the opiate antagonist naloxone (up to 4 mg/kg) were administered to normal volunteers. Dose-dependent increases in self ratings of tension-anxiety and anger-hostility were observed, supporting the hypothesized involvement of the endogenous opioid system in the modulation of human mood and feelings of well-being. Accompanying dose-dependent increases in systolic blood pressure and respiratory rate were found, suggesting that the lower doses of naloxone utilized in previous clinical studies were not sufficient to block the endogenous opioid system. CSF opioid activity in psychiatric patients and normals was measured using a sensitive radioreceptor assay developed by the authors. Results suggest diminished endogenous opioid system activity in some schizophrenics, and a relationship between opioid activity and state change in manic-depressive illness and anorexia nervosa. A complex but consistently observed relationship between ratings of anxiety and CSF opioid activity in normals and patients is consistent with basic science and clinical data suggesting interactions between CNS noradrenergic and opioid systems. General surgery was used as a strategy for studying the relationship of the endogenous opioid system to stress in humans; robust increases in levels of plasma beta endorphin immunoreactivity accompanying surgical stress and an inverse relationship between patient levels of plasma beta-endorphin immunoreactivity and postoperative analgesic requirement were observed. These data support the involvement of the endogenous opioid system in the human stress response and suggest that hormonal stress response and endogenous opioid system activity may relate to human endogenous analgesic mechanisms. PMID- 6295520 TI - [Activation of the fast sodium current through the membrane of single heart cells]. PMID- 6295519 TI - Biochemical and genetic investigations on gap junctions from mammalian cells. AB - Gap junction protein (26K) in mouse or rat liver has been studied using a rabbit antiserum directed against the sodium dodecylsulfate denatured 26K protein from mouse liver. The liver 26K protein has been localized in gap junction plaques of hepatic plasma membranes by immuno electron microscopy. Affinity purified anti 26K antiserum showed weak cross reactivity with mouse or bovine lens gap junction protein (MIP26). This result suggests some structural homology between the different gap junction proteins in liver and lens. After partial hepatectomy of young rats the liver 26K protein appears to be degraded and later resynthesized. A variant of established Chinese hamster fibroblastoid cells has been isolated and shown to be defective in metabolic cooperation via gap junctions. PMID- 6295521 TI - [Effect of the antineoplastic agent cis-Pt(NH3)2C12 and its isomer trans Pt(NH3)2C12 on the latent ATPase activity in isolated mitochondria of the rat liver and on the membrane-bound Na +,K + -ATPase activity in the bovine cerebral cortex]. PMID- 6295522 TI - [Fenibut binding with bicuculline-insensitive GABA receptors in the rat brain]. AB - (+/-) Fenibut beta-phenyl-GABA) was not able to displace 3H-GABA in Na+ independent GABA binding (IC50 greater than 250 microM). Nevertheless, (+/-) fenibut and (+/-) baclofen effectively displaced 3H-GABA in Ca2+ dependent GABA binding in the presence of 50 microM (+) bicuculline. (+/-) Fenibut was less potent in this respect. It is suggested that fenibut may act via bicuculline insensitive GABA receptors. PMID- 6295523 TI - [Isolation and study of the properties of the regulator subunit of cAMP-dependent protein kinase]. AB - The regulatory subunit of type II cAMP-dependent proteinkinase was isolated from cytosol of the rabbit small intestinal mucosa by affinity chromatography. The preparation contained 3 proteolytic enzymes and occurred in two forms differing as regards cAMP affinity. The cAMP-binding capacity of the preparation was equal to 17 nmol cAMP/mg protein. To study the topography of the cAMP-binding center, use was made of cAMP analogs. It was demonstrated that introduction of the substituents into the 8th position of the purine ring and substitution with respect to the N6-exoaminogroup affected insignificantly the analog affinity for the cAMP-binding center. At the same time the substituents introduced into the first position of the adenine base, into the area of the 2'-hydroxyl group of ribose and into the cyclophosphate part of the cAMP molecule considerably decreased the analog affinity for the regulatory center of type II cAMP-dependent proteinkinase. PMID- 6295525 TI - [Renin, kallikrein and angiotensin converting enzyme in human physical loading]. AB - The authors studied the correlation between the activity of renin, angiotensin converting enzyme, the activity of prekallikrein and the blood prekallikrein level in men performing veloergometric exercise. At the same time they recorded the hemodynamic parameters (systolic and diastolic arterial pressure, the systolic rate, stroke volume and cardiac index, specific peripheral resistance). The blood samples collected before and immediately after the exercise showed a 41.4%-increase in the activity of renin and a 95%-increase in that of kallikrein, whereas the level of prekallikrein and the activity of the converting enzyme declined by 19 and 13% (P less than 0.05). These changes were accompanied by an increase in the systolic rate (by 107%) and in the systolic arterial pressure (by 36.7%), as well as by a reduction of the specific peripheral resistance (by 41.4%). PMID- 6295524 TI - [3H-1-glutamate binding with the synaptic membranes isolated from the cerebral cortex and hippocampus of Krushinskii-Molodkina strain rats]. AB - Specific binding of 3H-L-glutamate to synaptic membranes isolated from the cerebral cortex and hippocamp of Wistar and Krushinsky-Molodkina (KM) rats examined both in a quiet awake state and after audiogenic seizures was compared. The dissociation constant (KD) values and binding capacity (Bmax) for KM rats did not differ significantly from the corresponding parameters of binding determined for Wistar rats (KD--89.8 +/- 18.1 and 102.6 +/- 12.5 nm, Bmax--1.23 +/- +/- 0.08 and 1.30 +/- 0.15 pmol/mg for the cortex and hippocamp, respectively). After audiogenic seizures the binding capacity of the hippocamp of KM rats was reduced by 30%. It is suggested that hippocampal glutamate receptors of KM rats are involved in the mechanism of convulsive activity formation. PMID- 6295526 TI - [Histochemical diagnosis of the neurons of cholinergic synaptic transmission]. AB - The authors studied neurons of the medulla oblongata of 5 human fetuses (22-27 weeks of development). Cholinacetyltransferase (CAT) activity was examined by the Berth method. Three neuronal types were diagnosed in the nuclei of the medulla oblongata with regard to CAT localization in the cytoplasm and synapses: (a) cholinergic-cholinoceptive neurons having CAT in the cytoplasm and in the innervating afferent fibers; (b) cholinergic-noncholinoceptive neurons with high CAT content, innervated with noncholinergic afferent fibers; (c) noncholinergic cholinoceptive neurons carrying cholinergic synapses. PMID- 6295527 TI - Lipid accumulation and production of colony-stimulating activity by the 266AD cell line derived from mouse bone marrow. AB - The availability of cloned lines of bone marrow stromal cells could facilitate the analysis of their role in hemopoietic cell development. The 266AD cell line was isolated from a colony of lipid-accumulating bone marrow cells growing in a collagen gel. 266AD cells have subsequently been maintained by passage in tissue culture plastic flasks about every 10 days for greater than 10 mo. Subconfluent cultures of cells are fibroblast-appearing, but in confluent cell sheets, prominent foci of lipid-containing cells develop in both uncloned and four separate cloned cell lines. Supernatants from confluent cultures containing lipid laden cells contain granulocyte-macrophage colony-stimulating activity (GM-CSA) for normal bone marrow cells and can induce differentiation of Abelson virus transformed murine promonocytic leukemia cells. 266AD cells were originally isolated in the presence of hydrocortisone, but hydrocortisone is not necessary for lipogenesis to occur. Growth of bone marrow cells in a collagen gel matrix provided a way to isolate stromal cells, and the 266AD cell line provides a means to examine the relationships between stromal cell lipogenesis and regulation of granulopoiesis. PMID- 6295528 TI - Non-contraceptive exogenous estrogens and risk of breast cancer: a review. AB - Results from epidemiologic and related studies of non-contraceptive estrogens and breast cancer are reviewed. Exogenous estrogens in high doses can enhance the risk of breast cancer. Moderate use of estrogens for menopausal symptoms probably has little effect on risk, but long-term users, and women who take high-strength preparations, appear to be at slightly increased risk. Exogenous estrogens probably reduce the protective effect of premenopausal oophorectomy, and may preferentially enhance the risk of breast cancer in women with some types of benign breast disease, although data from some studies do not support these conclusions. There is no evidence that the influence on risk of breast cancer is different for synthetic and conjugated estrogens. Needs for reanalysis of data from existing studies, and for additional investigations, are summarized. PMID- 6295529 TI - [Trophicity of digestive mucosae]. PMID- 6295530 TI - [Immunohistochemical localization of pituitary peptides in the rat pyloric antrum mucosa]. AB - Several peptides normally produced in the anterior pituitary lobe were searched in rat antral mucosa by immunocytochemistry. Peptides derived from pro opiomelanocortin were tested: some endocrine cells were immunoreactive with ACTH 17-39 antiserum, and only a few elements were stained with ACTH 1-24 and beta LPH antisera. No immunoreactive cells were observed using alpha MSH, beta MSH and beta endorphine antisera. Using an antiserum against beta endorphin, a few cells and nerve fibres were immunostained. The other pituitary hormones were also tested: numerous antral cells contained immunoreactive GH, and some cells immunoreactive PRL or compounds chemically related to these hormones. No cells were stained with antisera directed against glycoproteic hormones. This work showed that several peptides previously localized in the pituitary gland were found in the antral mucosa. Further studies are needed to identify the cell types containing these peptides and to determine their origin. PMID- 6295531 TI - Use of metabolic activation systems of tulip bulbs in the Ames test for environmental mutagens. PMID- 6295532 TI - Adsorption behavior of hydrogen sulfide inside micropores of Molecular Sieve Carbon 5A and Molecular sieve Zeolite 5A. PMID- 6295533 TI - [Oxidation of 2,4,5-triamino-6-hydroxypyrimidine by cytochrome C and 2,6 dichlorophenolindophenol]. PMID- 6295534 TI - Calmodulin: a multifunctional protein for the control of cellular activity and homeostasis. A review of its physico-chemical properties and pharmacological manipulation. PMID- 6295535 TI - Clinical evaluation of preoperative combined chemotherapy with neocarzinostatin (NCS) and 5-Fluorouracil (combined NF therapy) against gastric cancer. PMID- 6295536 TI - The "seagull" flap for syndactyly. AB - A technique is described for the treatment of post-burn syndactyly using a seagull shaped flap to produce a realistic commissure that does not subsequently advance distally. It can release volar digital contractures at the metacarpo phalangeal level and import skin with tactile "adherence" into the distal palmar area. PMID- 6295537 TI - Inhibition and recovery of the iodine uptake function of rat thyroids after 18 MeV proton irradiation in vivo. PMID- 6295539 TI - The effects of vegetable and animal protein diets on calcium, urate and oxalate excretion. AB - A group of 30 meat eating normal subjects were compared with a second group of vegetarians matched for age and sex. Their diets and urinary excretion patterns were compared by statistical analysis. A link between protein intake, particularly animal protein, and urinary calcium excretion was demonstrated and also that dietary calcium was inversely related to urinary oxalate excretion. Urinary oxalate increases with the vegetable protein content of the diet, but within the limits of these diets, animal protein does not affect oxalate excretion though it does affect excretion of urinary urate. PMID- 6295538 TI - Dietary management of urinary risk factors in renal stone formers. AB - Three hundred and ninety-two stone formers were investigated to exclude systemic disorders and to define the presence of haematological and urinary abnormality commonly associated with stone disease. Increased urinary excretion of calcium, oxalate or uric acid was found in 40% and there was more than one abnormality in 16% of the patients. The dietary habit of stone formers did not differ significantly from that of control subjects. Dietary advice to increase the consumption of fibre and reduce the consumption of sugar, refined carbohydrates and animal protein produced a significant reduction in the urinary excretion of calcium, oxalate and uric acid. We consider that reduction of the nutrient density of the diet by this means is the first line of management of idiopathic stone formers. PMID- 6295540 TI - Renal accumulation of ammonia: the cause of post-ischaemic functional loss and the "blue line". AB - The "blue line", a dark discoloration at the corticomedullary junction, is a constant finding after a significant period of renal ischaemia. In this study, it has been shown to be caused by packing of all of the peritubular capillaries at the corticomedullary junction with red blood cells. Five rats and 10 dogs were alkalinised by replacing their drinking water with 2% sodium bicarbonate for 2 weeks pre-operatively, in order to inhibit the glutaminase enzyme system and thereby decrease ammonia accumulation during ischaemia. Five rats and 6 dogs were used as unprotected controls. All animals were subjected to 60 min warm ischaemia. Renal function was significantly protected in the alkalinised rats (P less than 0.002) and dogs (P less than 0.001), with the serum creatinine rising to a maximum of 0.21 +/- 0.03 mmol/l in the alkalinised rats and 0.18 +/- 0.04 mmol/l in the alkalinised dogs. There was no "blue line" in the alkalinised animals. It is suggested that the "blue line" plays a central role in post ischaemic renal failure. Prevention of ammonia formation by alkalinisation protects against ischaemic renal damage and the formation of the "blue line". PMID- 6295541 TI - An in vitro physiological study of normal and unstable human detrusor muscle. AB - A systematic in vitro physiological study of isolated human detrusor muscle strips is reported. Detrusor strips were obtained at operation. Pre-operative urodynamic studies enabled specimens to be classified as normal or unstable. Comparisons were made between responses of normal and unstable detrusor strips. The results provide evidence of a previously undescribed feedback-loop operating via pre-synaptic alpha-adrenoceptors which controls detrusor contractility. Further evidence is provided suggesting that an imbalance in this system is present in unstable detrusor muscle. PMID- 6295542 TI - High serum vitamin B12 binding capacity as a marker of the fibrolamellar variant of hepatocellular carcinoma. PMID- 6295543 TI - Detection of rotavirus in handwashings of attendants of children with diarrhoea. PMID- 6295544 TI - Valproic acid and fetal abnormality. PMID- 6295545 TI - Selective changes in the density of beta 1-adrenergic receptors in rat striatum following chronic drug treatment and adrenalectomy. AB - The corpus striatum has a high density of beta-adrenergic receptors though it appears to contain low levels of beta-hydroxylated catecholamines. In an attempt to determine whether these receptors normally receive an endogenous input, the densities of beta 1 and beta 2-adrenergic receptors in rat caudate have been measured following adrenalectomy and after various pharmacological manipulations. Chronic administration of either pargyline, an inhibitor of monoamine oxidase activity, or desmethylimipramine, an inhibitor of norepinephrine uptake, resulted in a 20-25% decrease in the density of beta 1-adrenergic receptors while either adrenalectomy or the chronic administration of the non-selective beta-adrenergic receptor antagonist propranolol resulted in small but significant increases in the density of beta 1-receptors. These treatments did not lead to significant changes in the density of beta 2-receptors. It thus appears that the density of beta 1-receptors in the caudate is normally affected by changing levels of endogenous catecholamines. PMID- 6295546 TI - Extracellular calcium ionic activity in experimental spinal cord contusion. PMID- 6295547 TI - Effect of sympathectomy on extracellular potassium ionic activity and blood flow in experimental spinal cord contusion. PMID- 6295548 TI - GABA 'desensitization' of frog primary afferent fibers. AB - GABA (gamma-aminobutyric acid) depolarizes the terminals of primary afferent fibers of the in vitro hemisected frog spinal cord. During sustained or repetitive exposure to GABA or to muscimol, the amplitude of the depolarization is characterized by a rapid and exponential decline to a steady plateau level (desensitization). Desensitization to muscimol was eliminated by removal of Ca2+ and addition of Mn2+ to the superfusate--a finding consistent with the presence of 'receptor' ('true') desensitization (i.e., receptor inactivation). GABA desensitization was significantly reduced by exposure of the cord to either low Na+, low temperature, ouabain, dinitrophenol, (+/-)-nipecotic acid, or cis-1,3 aminocyclohexanecarboxylic acid. These treatments also significantly decreased the high affinity uptake of GABA when the latter process was studied by incubating frog spinal slices in Ringer's solution containing a low concentration of [3H]GABA. These results suggest that cellular transport processes can influence the form of GABA responses and indicate that neuronal removal of GABA is responsible in part for GABA desensitization. PMID- 6295549 TI - Effects of norepinephrine and serotonin upon spontaneous activity and responses to mossy fiber stimulation of CA3 neurons in hippocampal slices. AB - Effects of norepinephrine (NE) and 5-hydroxytryptamine (5-HT) upon spontaneous activity and responses to mossy fiber stimulation (mfs) were tested in 192 units of the field CA3 in the guinea pig and rat hippocampal slices. The drugs were added to the incubating medium or ejected by pressure from a micropipette. After NE superfusion firing rate increased in 52% of the reactive units, while activity of 48% was suppressed. The direction of the effect strongly correlated with pattern of spontaneous activity: only the cells with 'complex discharges' (short bursts of 2-4 spikes with attenuation of amplitude) were suppressed by NE; the cells with single spikes increased the level of activity. Similar excitatory effects of NE were observed in all units (n = 14) with single spike activity recorded in the field CA1. 5-HT increased activity in 30% of the reactive units and suppressed it in 70% of them. Some of the cells which were suppressed by 5 HT, were excited by NE. In more than a half of the units tested, 5-HT led to prolonged (up to 30-40 min) increase of the level of background activity irrespective of the initial excitatory or suppressive action; periodic grouped discharges appeared in some units under the influence of 5-HT. The response to mfs usually changed in the same direction as the level of background activity during application of NE and 5-HT, though some exceptions were observed in both cases. Prolonged (up to 30-40 min) facilitation of responses to mfs was present after application of 5-HT. PMID- 6295550 TI - Quantitative morphological analysis of spinal motoneurons. AB - Horseradish peroxidase was injected intracellularly into motoneurons responding to cutaneous stimulation of the central foot pad of the hind limb in cats. Three motoneurons were selected for detailed analysis: two excited by foot pad stimulation, and one postsynaptically inhibited by such stimulation. The overall lengths of the dendritic trees of the 3 cells ranged from 15.2 to 20.4 mm; the total surface areas ranged from 0.161 to 0.185 mm2. One cell had 9 primary dendrites, 39 terminal dendrites, and 69 dendritic branches in all. The second cell had 8 primary dendrites, 44 terminal dendrites, and 80 dendritic branches in all. For the third cell, these numbers were 14, 76 and 136. 75.2% of the total dendritic length of one of the cells was accounted for by branches of 3 of its 9 primary dendrites; for the second, 69.1% by 3 of 8; and for the third cell, 47.9% by 3 of 14. In contrast to the marked disparity in overall length of branches of the various primary dendrites, when the lengths of dendrites were analyzed by order of branching, dendritic branches of orders II-V each included greater than 15% of overall dendritic length. All 3 motoneurons displayed an exponential overall loss of total dendritic width with distance from the cell body, as well as an exponential decrease in Rall's dendritic trunk parameter. Four different patterns of branching were observed. Mean estimated electronic dendritic lengths were 1.3 for two cells, and 1.2 for the third. However, 13% to 28% of the dendrites of the 3 cells terminated more than two space constants from the cell body. In spite of striking differences among these cells in first-order morphology, on many of the detailed measures characterizing the dendritic trees of these motoneurons, they proved to be strikingly similar. PMID- 6295551 TI - Demonstration, characterization and localization of opioid binding sites in the midgut of the insect Leucophaea maderae (Blattaria). AB - The demonstration in the midgut of the insect Leucophaea of specific high affinity binding sites for a synthetic opioid represents the first report on neuropeptide binding in the digestive system of an invertebrate. Binding of the enkephalin analog DAMA (D-Ala2, Met5-enkephalinamide) is monophasic, saturable with respect to the concentration of the radioligand used, and stereospecific. Binding of the opiate antagonist naloxone to midgut homogenates is also monophasic, saturable, and stereospecific. The binding site density for DAMA is reduced by sodium and increased by manganese. By contrast, binding of naloxone is enhanced by sodium and unaffected by manganese. Lithium is equipotent with sodium in altering these values. Prolonged exposure of the organ to naloxone increases its binding capacity for DAMA. In midguts deprived of their autonomic innervation by severance of the recurrent nerve the binding capacity for the synthetic opioid is lower than in controls. Also, such 'denervated' organs are no longer affected by prolonged naloxone treatment. Results of tests for the presence in the midgut of non-peptidergic neurotransmitters (dopamine, norepinephrine) possibly operating in response to enkephalinergic signals, have thus far been negative. The results strongly suggest the existence in the digestive tract of this invertebrate of opioid receptors comparable to those in analogous mammalian systems. PMID- 6295552 TI - Opiate and non-opiate aspects of morphine induced seizures. AB - The intraperitoneal administration of morphine hydrochloride at doses of 300 mg/kg produced analgesia, catalepsy, and electrographic spiking in rats that developed into electrographic seizure patterns after approximately 2.5 h. Whereas naltrexone (12 mg/kg) reversed analgesia and catalepsy, and diminished electrographic spiking, it precipitated electrographic seizure activity similar to that observed following intraperitoneal morphine alone. These seizures were accompanied by behavioral convulsions. No tolerance to these seizures developed with repeated paired administration of morphine and naltrexone or in morphine tolerant rats, but rather potentiation was observed. The epileptogenic effects were found to be potentiated in amygdaloid kindled rats, as well. It was concluded that morphine at these doses activates two different epileptogenic mechanisms, one mediated by opiate receptors, the other not. The possibility of the simultaneous activation of a morphine sensitive anticonvulsant mechanism is discussed. PMID- 6295553 TI - Intracellular protein changes during pentylenetetrazole induced bursting activity in snail neurons. AB - The intracellular protein changes during pentylenetetrazole (PTZ)-induced bursting activity (BA) which is characteristic of seizure discharge were investigated using microdisk electrophoresis with 5 identified neurons. The identified neurons of the snail, Euhadra peliomphala, were used. The PTZ sensitive neurons which manifest marked BA by application of PTZ were examined. PTZ induced in PTZ-sensitive neurons: (1) a prominent increase of 5-7 kdalton protein and (2) peak separation into 3 peaks of 10-15 kdalton protein. In the 5-7 and 10-15 kdalton protein, a marked increase in radioactivity of 45Ca was observed after PTZ application. PTZ-non-sensitive neurons showed neither these protein changes nor 45Ca incorporation into these proteins. The above findings suggest that during PTZ-induced BA, intracellular protein changes occurred in relation to the intracellular calcium shift. PMID- 6295554 TI - Atropine enhances nicotinic cholinergic EPSPs in rat neostriatal slices. PMID- 6295555 TI - The effect of dantrolene on tetanic potentiation of MEPP frequency in EGTA containing salines. AB - The effect of dantrolene on tetanic potentiation of MEPP frequency in salines containing 1 mM EGTA and no added Ca2+ has been examined. Dantrolene pre treatment results in no significant change in the response of normalized MEPP frequency to tetanus at stimulation rates of both 25 and 100 Hz. This result is compared to the way in which dantrolene interacts with other agents that modify spontaneous release and it is suggested that intracellular Ca2+ movements during nerve tetanus may well be complex involving more than one Ca2+ store. PMID- 6295556 TI - Effects of intraocular injections of 6-hydroxydopamine on dopamine-dependent cyclic AMP accumulation in intact pieces of carp retina. AB - Dopamine-stimulated cyclic AMP accumulation was measured in intact pieces of carp retina following intraocular injections of the dopaminergic neurotoxin 6 hydroxydopamine (6-OHDA). This treatment is known to induce a selective destruction of dopaminergic nerve terminals, and in these experiments caused a 94% reduction in retinal dopamine content. However, dopamine-dependent cyclic AMP accumulation was essentially unaltered in retinas exposed to 6-OHDA, indicating that the dopamine receptors linked to adenylate cyclase in the carp retina are located mainly on postsynaptic elements, and not presynaptically on the dopaminergic terminals. PMID- 6295557 TI - Lithium: effects of short-term and chronic treatments in rats on the activity of dopamine-beta-hydroxylase (DBH) in the central versus peripheral nervous system. AB - The therapeutic usefulness of lithium in mania is now well-established. However, the precise neurochemical mechanisms by which lithium brings about its effects remain poorly understood. This report describes the effects of lithium on the central and peripheral levels of activity of dopamine-beta-hydroxylase (DBH), the enzyme that catalyzes the final step in the biosynthetic pathway to the formation of norepinephrine (NE) from dopamine. Since the neurotransmitter NE has been implicated in the mediation of lithium's action, what effects lithium may have on the activity of DBH was deemed worth investigating. Lithium chloride was administered intraperitoneally in rats on a short-term and chronic basis and the effects on DBH activities in the hypothalamus and serum were evaluated. A significant reduction in DBH activity occurred in the hypothalamus after short term lithium but there was no change following chronic treatment. In contrast, serum DBH activity remained unchanged after both acute and chronic lithium injection. Since increased noradrenergic activity in the brain has been implicated in the etiology of mania, the lithium-induced decrease in hypothalamic DBH is suggestive of a neurochemical mechanism by which lithium may act in psychotic patients. PMID- 6295558 TI - Homeostatic control of ascorbate concentration in CNS extracellular fluid. PMID- 6295559 TI - Lack of correlation between physiological and morphological features of regenerating frog neuromuscular junctions. AB - Morphological, ultrastructural and electrophysiological properties of endplates in the frog cutaneous pectoris muscle were monitored at various times after nerve crush. The lengths of regenerating terminals were measured in nitroblue tetrazolium-stained material. Quantal content and the frequency of spontaneous potentials were obtained from intracellular records, and they were correlated to individual terminal lengths in order to normalize their values to a standard unit of terminal length. By day 16 after nerve crush, terminal length reached a plateau near 90% of normal. In contrast, quantal content per unit terminal length quickly recovered to 67% of normal by 7-12 days but did not increase thereafter. The frequency of spontaneous potentials per unit terminal length returned much more slowly, as it remained low (near 18% of normal) throughout the entire first month before recovering to normal levels later on. Ultrastructural observations on terminal size, densities of mitochondria and synaptic vesicles, and multiple innervation could not explain these different recovery rates. It is concluded that terminal length, spontaneous transmitter release and evoked transmitter release, are not restored in parallel after reinnervation. PMID- 6295560 TI - Comparison of morphology and physiology of synapses formed at ectopic and original endplate sites in frog muscle. AB - The properties of junctions formed de novo in ectopic regions of muscle were studied and compared to those of endplates reinnervated after nerve crush and to normals. The nerve to the frog cutaneous pectoris muscle was transposed and implanted onto an endplate-free (ectopic) region of that muscle, while the endplate-containing portion was excised. New synapses, which subsequently formed in the remaining muscle segment, were studied morphologically and physiologically at postoperative times from approximately 3 weeks to a year. Junctions formed de novo consisted of numerous small contacts, and were only 15% as long as those in control groups. Yet, quantal contents were 2.5-4.7 times larger at de novo junctions. PMID- 6295561 TI - Convergence of muscle spindle afferents on single neurons of the cat dorsal spino cerebellar tract and their synaptic efficacy. AB - By means of tungsten microelectrodes, action potentials from axons within the dorsal spino-cerebellar tract (DSCT) and from muscle spindle afferents were recorded. A quantitative study was performed in monomuscular DSCT neurons which were excited predominantly by Ia fibers originating in the gastrocnemius muscles. In some experiments single Ia fibers were stimulated electrically while the impulse sequence of a DSCT neuron postsynaptic to the respective afferent fiber was recorded. The gastrocnemius DSCT neurons receive excitatory inputs from 10-18 Ia muscle spindle afferents. The efficacy of each of these inputs is very similar. Thus the neuronal activation decreased approximately linearly with the number of the excitatory afferents cut. Cross-correlograms between the impulse sequence of a Ia gastrocnemius muscle spindle afferent and a DSCT neuron postsynaptic to it exhibited an increased discharge probability of the DSCT neuron from 3-4 ms to 10 ms after the Ia action potential. With increasing impulse rates of the Ia afferent fibers, the excitatory efficacy of the single action potential decreased, but the overall excitation increased with the presynaptic discharge frequency, according to a hyperbolic function. This effect was tested by electrical stimulation of a single Ia axon exciting the DSCT neuron recorded. Interval histograms computed from DSCT neuron impulse trains at steady stretch conditions were predominantly monomodal. They can be well approximated by a Gaussian distribution. The coefficient of variation was independent of the mean activity. At impulse rates above 25 imp X s-1 a negative correlation between successive intervals was observed in first order joint interval diagrams. With an increasing mean discharge rate this correlation (expressed as the serial linear correlation coefficient of the first order r1,2) became stronger up to--0.62 at 90 imp X s-1. Only in a few neurons did the higher order linear correlation coefficients deviate significantly from zero. In 15% of the observed histograms double discharging (mean interval 3-5 ms) produced bimodal distributions. Under steady-state conditions the response of Ia-activated DSCT cells are linearly related to muscle stretch within a middle range of extensions. The differences between Ia impulse pattern and DSCT neuron impulse pattern at steady stretch are discussed. The number of large dendrites of the principal cells in the nucleus dorsalis (Clarke's column) corresponds to the number of excitatory afferent muscle fibers. It is assumed that each excitatory Ia axon sends one axon collateral to the DSCT neuron, forming a climbing type terminal mainly on one of the large dendrites of a DSCT cell. PMID- 6295562 TI - Amino acids as taste stimuli. II. Quality coding. AB - Two experiments were performed in rats to evaluate the relative taste qualities of 12 L-amino acids, each at a concentration which evoked half the maximum response for that chemical. The first study involved recording the activity of 40 individual chorda tympani fibers to the stimulus series. Only 34% of the evoked responses resembled the short latency phasic-tonic activity which characterizes gustatory responses to inorganic salts and acids. 32% had latencies exceeding 1 s; another 27% consisted of only a phasic burst lasting less than 1 s. The remaining 7% were inhibitory. Both long latency and purely phasic activity were stimulus selective: 61% of the former were in response to Gly or Pro while 69% of the latter were evoked by Cys-HCl, Lys-HCl or His. Response inhibition was not associated with either specific fibers or stimuli. Thus amino acids, which to humans represent a class of perceptually complex stimuli, show a corresponding complexity of evoked neural properties in the rat. The second study employed a conditioned taste aversion paradigm to assess the qualitative similarity of each amino acid to the others and to the 4 prototypical taste stimuli; NaCl, HCl, quinine-HCl and sucrose. Some amino acids showed strong generalization to a single gustatory prototype (Pro and Gly to sucrose; Cys-HCl to HCl); others generalized well to multiple prototypes (e.g. Arg to sucrose and NaCl). Several showed poor generalization to all 4 prototypical tastes, calling into question the assumption that these 4 totally encompass the gustatory domain. PMID- 6295563 TI - Ontogenesis of opiate binding sites and radioimmunoassayable beta-endorphin and enkephalin in regions of rat brain. AB - The postnatal changes in the levels of radioimmunoassayable enkephalin and beta endorphin, as well as the densities of [3H]methionine-enkephalin and [3H]naloxone binding sites in rat cerebellum, brainstem and whole forebrain were determined. The opiate peptides and the opiate binding sites reached their highest levels at the first week postpartum in the cerebellum, at the second week in the brainstem and at the third week in the whole forebrain. This finding is in line with the developmental profiles of other well-established neuronal pathways which also showed a caudal-to-rostral sequence of development. Moreover, there was a close relationship between the elevation and decline in the amounts of opiate binding sites and in the levels of opiate peptides in each brain region. These observations are consistent with other evidence which suggests that enkephalin and beta-endorphin are functioning as neurotransmitters or neuromodulators in the central nervous system. PMID- 6295564 TI - Immunoreactive beta-endorphin in the hypothalamus of female rats: changes in content and release during prepubertal development. AB - Female Wistar rats of different ages (1-45 days) were used. Extracts were made of the mediobasal hypothalamus (MBH) and beta-endorphin immunoreactivity (beta-ENDi) was quantitated by radioimmunoassay. Low but significant amounts of beta-ENDi (6.5 ng/MBH) were present on the first postnatal day. Hypothalamic beta-ENDi content did not change during the first week but decreased during the second week to a minimum (4.5 ng/MBH) on day 14. Thereafter, beta-ENDi increased rapidly to 13 ng/MBH on day 28 and remained at this level. Gel filtration showed that beta ENDi substances with chromatographic characteristics identical to those of beta END and beta-LPH were present in MBHs of 14-, 20- and 45-day-old rats. A beta ENDi substance, possibly representing beta-END1-27, was nearly absent on day 14, but represented a major component of the MBH of the 45-day-old rat. In vitro incubation of MBH resulted in spontaneous release of beta-ENDi. Depolarization of neuronal membranes by incubation in medium containing 45 mMK+ stimulated beta ENDi release. Both the spontaneous and K+-stimulated release of beta-ENDi were low on day 10 but reached postpubertal levels on day 20. These observations lead us to propose that the beta-ENDi-containing neurons in the hypothalamus of developing female rats rapidly mature between 14 and 20 days after birth. This may be causally related to the rapid decrease in circulating FSH levels that occurs during this period. PMID- 6295565 TI - [Ontogenesis of pro-opio-melano-cortin neurons in the rat]. AB - Immunocytochemical study of the pro-opio-melano-cortin neurones in rat fetuses and newborns indicates that: 1) they appear precociously in the mediobasal hypothalamus, as early as day 13 of fetal development, that is to say 3 days before pro-opio-melano-cortin-containing pituitary cells; 2) between day 13 and day 17 they display synchronous changes in their intracellular patterns of immunoreactivity, arguing in favour of their cyclic secreting activity, which precedes the establishing of their projections; 3) beginning with day 18 until birth, the somatal immunoreactivity disappears transitorily; during the same period and the first postnatal days, their immunoreactive fibres extend in various brain regions. PMID- 6295566 TI - [Cytochemical detection and cytofluorimetric analysis of alkaline phosphatase in continuous lines of thymic lymphomas induced in mice by a radiation leukemia virus]. AB - Alkaline phosphatase was studied in cell lines established from Radiation Leukemia Virus induced thymic lymphomas of C57BL/Ka mice. The cytochemical staining techniques and flow cytofluorimetry analysis described by Dolbeare et al. (J. Histochem. Cytochem., 1980, 28, 419-426) were used. The alkaline phosphatase found in lymphoma cells was heat labile and L-homoarginine. L phenylalanine and p-bromotetramisole sensitive and is probably similar to the isoenzyme present in mouse placenta, kidney and liver. Very little alkaline phosphatase activity was detected in normal thymus of adult mice, suggesting that the method used in the paper could be helpful for studying the emergence of the first neoplastic cells during the leukemogenic process. PMID- 6295567 TI - [Increase, in extracts of cancerous or transformed cells, of protein kinase activity phosphorylating tyrosine]. AB - A protein kinase activity which phosphorylates tyrosine in the presence of Mn++ ions has been detected among different protein kinase activities in a postnuclear cell fraction sedimenting mostly at 12000 X g. The proportion of phosphotyrosine relative to phosphoserine and phosphothreonine is at a low level in normal embryonic cultured cells and is increased several fold in various types of cells either malignant or transformed in vitro. This proportion which may reach up to 8% is in the same range as that found in extracts of avian cells transformed by Rous sarcoma virus, where the viral kinase P60 src is expressed. In two ascitic tumour cells the tyrosine kinase activity seems to be associated with the mitochondrial fraction. PMID- 6295568 TI - [Polymorphism of HLA genes: I. Demonstration of a close correlation between DNA fragments determined by BglI restriction enzyme and HLA class I antigens]. AB - Description of DNA fragments associated to HLA class I gene is possible by using restriction enzymes which determine these fragments and specific DNA probes which permit their detection. In one family, with a child presenting a recombination between HLA-A and C, six fragments determined by the enzyme BglI were found to be polymorphic. The informative fragments segregate with HLA, either with a whole haplotype or with one of the two recombinant segments of the HLA complex. In a small sample of population they correlate with one (A11) or with a group of known cross-reactive antigens serologically defined (A3 and A11; A25 and A26. Another fragment is associated with unknown cross-reactive antigens (A2 and A29). PMID- 6295569 TI - Gestational trophoblastic neoplasia. PMID- 6295570 TI - [Selective angiography in the diagnosis of insulinoma]. PMID- 6295571 TI - Effect of cholecalciferol derivatives on the mechanical properties of chick bones. AB - Chicks were depleted of vitamin D, divided into groups, and treated daily with (a) cholecalciferol, (b) 1 alpha-hydroxycholecalciferol [1 alpha (OH)-D3], (c) 24R, 25-dihydroxycholecalciferol [24R,25-(OH)2D3], or (d) 1 alpha (OH)D3 and 24R,25(OH)2D3. Two additional groups of chicks were studied, one that was continuously depleted of vitamin D, and another that was continuously supplemented with the vitamin, since day 1. After killing, the tibiae were removed and tested for their mechanical properties. Bending load was applied to the midshaft, and the intrinsic properties of this site, its quantity and geometry were analyzed. From a mechanical point of view, the weakest bones found were of birds depleted of vitamin D, whereas the strongest were of those treated with 1 alpha (OH)D3. Only the bones of the 24R,25(OH)2D3-treated or the 1 alpha (OH)D3 and 24R,25(OH)2D3-treated groups of birds showed mechanical properties comparable to those obtained with vitamin D-replete chicks. PMID- 6295572 TI - Chemical and biochemical aspects of superoxide radicals and related species of activated oxygen. AB - The spectrum of biological processes in which oxygen is used by living systems is quite large, and the products include some damaging species of activated oxygen, particularly the superoxide radical (O-.2) and hydrogen peroxide (H2O2). Superoxide radicals and hydrogen peroxide, in turn, can lead to the formation of other damaging species: hydroxyl radicals (.OH) and singlet oxygen (1O2). Hydroxyl radicals react with organic compounds to give secondary free radicals that, in the presence of oxygen, yield peroxy radicals, peroxides, and hydroperoxides. Formation, interconversion, and reactivity of O-.2 and related activated oxygen species, methods available for their detection, and the basis of their biological toxicity are briefly reviewed. PMID- 6295573 TI - The pathophysiology of superoxide: roles in inflammation and ischemia. AB - The superoxide radical plays major roles in the neutrophil-medicated acute inflammatory response and in postischemic tissue injury, although the sources and actions of the radical are quite different in these two pathological states. While neutrophils produce superoxide for the primary purpose of aiding in the killing of ingested microbes, a second useful function has evolved. The superoxide released from actively phagocytosing neutrophils serves to attract more neutrophils by reacting with, and activating, a latent chemotactic factor present in plasma. Superoxide dismutase, by preventing the activation of this superoxide-dependent chemotactic factor, exerts potent anti-inflammatory action. During ischemia, energy-starved tissues catabolize ATP to hypoxanthine. Calcium transients in these cells appear to activate a calmodulin regulated protease which attacks the enzyme xanthine dehydrogenase, converting it to a xanthine oxidase capable of superoxide generation. When the tissue is reperfused and reoxygenated, all the necessary components are present (xanthine oxidase, hypoxanthine, and oxygen) to produce a burst of superoxide which results in extensive tissue damage. Ischemic tissues are protected by superoxide dismutase or allupurinol, an inhibitor of xanthine oxidase. PMID- 6295574 TI - The enzymatic basis for O-.2 production by human neutrophils. AB - O-.2 production is the first step in the generation of a group of powerful microbicidal oxidants by neutrophils. The production of O-.2 is catalyzed by a membrane-bound, NADPH-preferring flavoprotein oxidase, a conclusion supported by much evidence including the discovery of a new form of chronic granulomatous disease caused by a mutation affecting that oxidase directly. Also involved in the O-.2-forming reaction is a b-type cytochrome; the role of this cytochrome is as yet undefined, though it does not appear to be on the direct route of electron transfer between NADPH and oxygen. It has been postulated that quinones too participate in the O-.2-forming reaction, but further work is necessary to define their role more fully. PMID- 6295575 TI - Factors which affect DNA strand breakage in human leukocytes exposed to a tumor promoter, phorbol myristate acetate. AB - We have recently reported that phorbol myristate acetate (PMA) induces extensive DNA strand break damage in human peripheral blood leukocytes. The mechanism of action involves superoxide anion and hydrogen peroxide which are generated by phagocytes during the "respiratory burst." In this report, we describe the effect of various inhibitors and scavengers on PMA-induced DNA damage. Azide and cyanide greatly increased the level of damage; sulfhydryl compounds (glutathione, cysteine, and cysteamine) and ascorbate markedly decreased the level of damage. Hydroxyl radical scavengers such as dimethyl sulfoxide (DMSO) and glycerol also decreased the level of damage but apparently did so by inhibiting the respiratory burst. Diethyldithiocarbamate (DDC) increased the level of DNA damage at low concentrations (less than 1 mM), but decreased DNA damage at greater than or equal to 1 mM. The results are consistent with a mechanism involving superoxide anion and hydrogen peroxide, but the precise reaction (free radical or enzymatic) responsible for DNA strand breakage has not been determined. The PMA-stimulated phagocyte is an interesting model system for looking at "active oxygen" mediated DNA damage and factors which influence it. PMID- 6295576 TI - Role of superoxide anion radicals in microvascular permeability and leukocyte behaviour. AB - Inflammation is associated with the accumulation and activation of phagocytic cells, such as polymorphonuclear leukocytes, and with the subsequent release and generation of a group of activated oxygen species, some of which are free radicals. These studies were carried out to assess the influence of enzymatically generated free radicals on both microvascular permeability and leukocyte adhesion. The extravasation of fluorescein-labelled dextran mean molecular weight (MW) 150 000 was used to assess microvascular permeability and methodology was developed to measure in vivo leukocyte endothelial interactions. Enzymatically generated superoxide anion radical (O-.2) was associated with an increase in macromolecular extravasation (seen primarily from postcapillary venules) and an increase in leukocyte adhesion. Macromolecular extravasation was found to be dependent on the generation of a hydroxyl radical related interaction while leukocyte adhesion was dependent on the presence of O-.2. It is suggested that the permeability alterations and increased polymorphonuclear leukocyte adhesion seen during inflammation may be partially related to the release of free radicals from inflammatory cells. PMID- 6295577 TI - Active oxygen and medicine. Concluding remarks: a prospective view of active oxygen in medicine. AB - Factors are identified that influence the initiation, propagation, and efficiency of free-radical processes, or that ameliorate or mask them. Major biochemical disruptions of the ischemic myocardium or the central nervous system are cited for their underlying free-radical reactions. Encouraging attempts at rational therapy of disease having a free-radical component are referenced. PMID- 6295578 TI - The toxic effects of ouabain: a voltage-clamp study. AB - The electrophysiologic effects of a toxic concentration of ouabain (10(-5) M) were studied in frog atrial trabeculae. The toxic concentration was determined by the appearance of a negative inotropic effect and an increase in basal tension. Current- and voltage-clamp measurements were performed. Ouabain did not alter the passive electrical properties of the preparation. Under current-clamp conditions the membrane depolarized and the action potential amplitude as well as its maximum rate of rise decreased. The current--voltage curve for the fast inward current was shifted toward more positive potentials and the maximum sodium current decreased. The maximum sodium conductance was also reduced. The process of reactivation of the fast inward current was accelerated. The slow inward current and the maximum slow conductance also decreased under ouabain. These effects could explain the negative inotropic action of high concentrations of glycosides, as well as the action potential changes observed by several investigators. They also help to understand the arrhythmogenic effects of high concentrations of digitalis. PMID- 6295579 TI - Chronic alcohol feeding and its withdrawal on the structure and function of the rat liver plasma membrane: a study with 125I-labelled glucagon binding as a metabolic probe. AB - The effect of chronic alcohol administration on the structure and function of the rat liver plasma membranes has been investigated. Chronic alcohol administration did not affect the yield of these membranes using conventional isolation procedures. The extent of plasma membrane enrichment or contamination with other interior membranes was identical in the control and alcoholic preparations. The binding of 125I-labelled glucagon to these experimental liver plasma membranes was significantly decreased. Scatchard analysis of the high affinity sites showed a significant reduction (approximately equal to 35%) in receptor number rather than binding affinity, which was not altered. This anomaly persisted through 72-h withdrawal of alcohol. These data suggest that very stable changes were induced in these liver plasma membranes after prolonged alcohol ingestion. PMID- 6295580 TI - Slow inward Ca current in frog heart: theoretical evidence against a voltage dependent inactivation. AB - The validity of a Hodgkin-Huxley type voltage-dependent inactivation of slow inward Ca current (Isi) was tested in frog heart using a computer simulation. The time course of Isi was calculated during the development of a frog atrial action potential (AP). With a time constant of inactivation (tauf) of 55 ms at a membrane potential (Em) of -15 mV, the variation of Isi was biphasic: after a transient increase followed by a decrease to zero, Isi partially "reactivated" (at the beginning of the AP repolarization phase) and then fully deactivated. The "reactivation" phase of Isi developed whether tauf was an increasing, decreasing, U-shaped, or bell-shaped function of Em. The addition of an independent and slower process responsible for the recovery from inactivation only partly suppressed the "reactivation" phase. However, until now there was no experimental evidence supporting such a biphasic variation of Isi during AP repolarization. Thus our results indicate that the Hodgkin-Huxley type model of the voltage dependence of Isi-inactivation process may not correctly represent the actual behavior of frog cardiac muscle. PMID- 6295581 TI - The effects of indomethacin on uterine activity and prostaglandin (PG) concentrations during labor induced by administering ACTH to fetal sheep. PMID- 6295582 TI - Effect of the carbon source and cyclic AMP on isocitrate dehydrogenase, succinate dehydrogenase, and malate dehydrogenase in Klebsiella pneumoniae C3. AB - When strain C3 of Klebsiella pneumoniae is grown on a minimal medium with excess glucose, isocitrate dehydrogenase, malate dehydrogenase, and succinate dehydrogenase specific activities increase in the last period of the exponential growth phase and in the beginning of the stationary phase. Glucose exhaustion does not alter the development of malate dehydrogenase and succinate dehydrogenase, but specific activities are higher than those obtained with excess glucose. In contrast, glucose exhaustion can be correlated with a decrease of isocitrate dehydrogenase specific activity in the stationary phase. Induction of strain C3 isocitrate dehydrogenase by glucose in complex medium and repression by cAMP in mineral medium were observed. Glucose induction and the NADP/NADPH ratio are suggested as regulatory mechanisms controlling isocitrate dehydrogenase synthesis in the Enterobacteriaceae, but the former appears to be restricted to some Klebsiella strains. PMID- 6295583 TI - A comparison of the effects of several antifungal imidazole derivatives and polyenes on Candida albicans: an ultrastructural study by scanning electron microscopy. AB - The early events in the interaction of two polyene (amphotericin B and nystatin) and five imidazole (clotrimazole, ketoconazole, miconazole, isoconazole, and econazole) antimycotics used at fungicidal concentrations with the surface of Candida albicans were studied by scanning electron microscopic examination of treated intact young yeast cells, treated spheroplasts, and spheroplasts liberated from treated young yeast cells. In all cases, treatment lasted 2 h. The polyenes passed through the yeast cell wall and interacted with the cytoplasmic membrane causing the spheroplasts to lose their characteristic spheric form and to liberate their contents. Clotrimazole caused the formation of numerous circular openings in the cytoplasmic membrane, but only when the agent was used to treat spheroplasts directly. Ketoconazole, miconazole, isoconazole, and econazole interacted with the cell wall causing formation of convolutions and wrinkles. The three imidazole derivatives that are structurally closely related, miconazole, isoconazole, and econazole, inhibited the enzyme-catalyzed release of spheroplasts from young yeast cells. PMID- 6295584 TI - Presidential address: XVII Canadian Congress of Neurological Sciences. Cushing's disease: 50 years later. AB - The recent improvements in transsphenoidal microsurgery have once again placed the emphasis on the basophilic adenoma of the pituitary as the etiology of the disease originally described by Harvey Cushing fifty years ago. In 75 operated cases, tumors were found in 60 cases: 52 were microadenomas, 4 were larger adenomas and 4 were invasive. In 15 cases, no tumor was found at surgery; a selective partial central hypophysectomy was performed in 11 cases and a total hypophysectomy in 4. Among the latter, 3 additional microadenomas were found at pathology. The majority of the microadenomas were located in the center of the gland most often near or within the neural lobe. In 59 patients with a non invasive tumor, initial cure was obtained in 52 (88%); only 1 of the 4 patients with an invasive tumor was cured. In the 12 cases without surgical or pathological tumor, partial or total hypophysectomy resulted in 10 cures (83%). PMID- 6295585 TI - Myocardial contusion in chest trauma. AB - Because myocardial dysfunction may result from severe trauma, the author assessed, prospectively, left and right ventricular function in 25 patients who had sustained severe trauma, including blunt chest injury, by electrocardiographically gated blood pool radionuclide angiography. Focal abnormalities of ventricular wall motion were defined in 17 patients: right ventricular in 12, left ventricular in 2 and biventricular in 3. Traumatic tricuspid insufficiency demonstrated in two patients was subsequently verified by contrast angiography. Other means of detecting myocardial contusion (enzymatic, electrocardiographic and scintigraphic) proved to be insensitive when compared with radionuclide angiography. Two of the five deaths were attributed to refractory arrhythmias. Surgical or autopsy evidence of traumatic myocardial injury was obtained in five instances when radionuclide angiography indicated contusion. Of the 13 patients available for follow-up, 11 showed complete or partial resolution of the ventricular wall abnormality and in 2 there was no change. Comprehensive cardiopulmonary monitoring revealed an inverse relation between the right ventricular ejection fraction and pulmonary vascular resistance as well as between the pulmonary vascular resistance and left ventricular ejection fraction and left ventricular end-diastolic volume. Further, as the right ventricular end-diastolic volume was increased in trauma, left ventricular function and compliance were reduced. In blunt chest trauma, right ventricular contusion occurs more frequently than has been recognized previously and positive radionuclide angiography constitutes prima facie evidence of direct myocardial injury. Moreover, left ventricular function remains preload-dependent, but may be depressed by elevated pulmonary vascular resistance, impeding the blood flow from the right to left ventricle, and decreased left ventricular compliance, or both. PMID- 6295586 TI - Cricopharyngeal myotomy. AB - Cricopharyngeal myotomy has been used in the treatment of pharyngoesophageal diverticulum and various neurogenic, myogenic and myoneurogenic disorders. An appreciable number of patients with pseudobulbar palsy due to cerebrovascular accidents and patients with idiopathic hypertrophy of the cricopharyngeal muscle will greatly benefit from this procedure. This paper describes the indications for, and results of, 20 consecutive cricopharyngeal myotomies carried out with or without diverticulectomy. All 20 patients experienced cervical esophageal dysphagia and 55% had substantial weight loss. The most valuable investigation is roentgenography of the pharynx and esophagus, which will confirm megapharynx, hypopharyngeal stasis, weak or absent pharyngeal contractions and regurgitation. Hypertrophic cricopharyngeal muscle was demonstrated in 9 of the 20 patients. The diagnostic value of endoscopy and esophageal manometry is limited. The results were considered excellent in all patients with pharyngoesophageal diverticulum and idiopathic hypertrophy of the cricopharyngeal muscle. Marked symptomatic and objective improvement was achieved in patients with cerebrovascular accidents, vagal injuries and amyotrophic lateral sclerosis. However, the result was poor in patients with myoneurogenic disorders. PMID- 6295587 TI - The pathogenesis of neuropsychiatric lupus. AB - Neuropsychiatric lupus is a frequent and serious complication of systemic lupus erythematosus. Although its pathogenesis has not been fully elucidated several mechanisms based on findings in animals and humans have been proposed: cerebral vasculitis; the cross-reaction of lymphocytotoxic antibodies with brain tissue; choroid plexus dysfunction; and the blocking of neurotransmission by noncytotoxic antibodies. Any or all of these mechanisms may be active in a particular patient at a given time. PMID- 6295589 TI - Small cell undifferentiated carcinoma of the larynx. Report of two patients and review of 13 additional cases. AB - Two patients with small cell undifferentiated carcinoma of the larynx (SCUCL) are described and 13 additional well-documented cases are reviewed. SCUCL affects middle-aged and elderly smokers and has a 2:1 male predominance. Half the patients presented with cervical metastases, an additional 43% developed regional metastases, and 71% ultimately had distant metastases. Thyroid gland involvement occurred in 29% of cases. Ten of fourteen patients had died of their neoplasms (mean survival, 7.8 months). One was living with unresectable tumor. Three were clinically disease-free, 30, 15, and six months, respectively, after diagnosis. Following initial therapy, six patients had recurrent or persistent local tumor or regional metastases. None of them received both total laryngectomy and ipsilateral cervical lymphadenectomy. Four of the six patients had radiation, suggesting that radiotherapy was not always effective in the control of local or regional disease when used in lieu of more extensive surgery. Three patients received chemotherapy, in addition to other modes of treatment, and their mean survival compared favorably with the overall group. The tumor from one of the patients contained a squamous component, and, ultrastructurally, both neoplasms had squamous and neuroendocrine features. PMID- 6295588 TI - Multiple malignant neoplasms 40 years after angiography with Thorotrast. PMID- 6295590 TI - CEA-like material in cytosols from human breast carcinomas. Correlation with biochemical and pathologic parameters. AB - CEA-like material was found in 51 of 62 primary human breast carcinomas and in only 2 of 12 fibroadenomas. Levels of carcinoma CEA-like material correlated weakly with cytoplasmic estradiol receptor levels, total cytosol estrogens, and cytosol progesterone. Levels of CEA-like material showed no significant correlation with carcinoma stage, grade, cellularity, size or histologic type. Levels, however, correlated inversely with lymphocyte infiltration. PMID- 6295591 TI - The fine needle aspiration cytology of mediastinal lesions. AB - Fine needle aspiration cytology was performed in 19 mediastinal lesions. Of seven malignant neoplasms six were correctly diagnosed as malignant and in five of these accurate tumor classification was possible. Of the 12 benign lesions only four cases, all thymomas, could be diagnosed cytologically. No false-positive diagnoses of malignancy were made. The only complication of the procedure was minor pneumothorax in two patients. In five cases the use of the technique spared the patient from more invasive diagnostic procedures; in several others, valuable information was obtained prior to surgery. PMID- 6295593 TI - Adjuvant chemotherapy of malignant fibrous histiocytoma of bone. AB - Malignant fibrous histiocytoma (MFH) is a pleomorphic sarcoma that is uncommon in children. It most frequently arises from the soft tissues; however, it has been recently established that primary bone MFH also exists. Surgical resection or amputation is the cornerstone of treatment for MFH of bone. But, with this modality of therapy alone the majority of patients develop either distant metastases or local recurrence. This study reports on three adolescent girls with MFH of bone who were successfully treated with radical resection and 18 months of adjuvant chemotherapy with vincristine, high dose methotrexate, Citrovorum Factor rescue, and Adriamycin. All three patients remain disease-free for a follow-up period of 42-48 months. The current regimen was well tolerated. Morbidity was minimal, with no patient developing any significant drug-related complications. The adjuvant chemotherapy regimen described appears to be effective in prolonging survival in patients with MFH of bone and appears to warrant further study in additional patients. PMID- 6295592 TI - Management of the syndrome of inappropriate antidiuretic hormone secretion in small cell lung cancer. AB - From 1976 to 1980, 18 of the 250 patients (7%) seen with small cell carcinoma of the lung had clinically evident inappropriate secretion of antidiuretic hormone (ADH). Hyponatremia was usually severe (116 +/- 7 meq/l), and eight patients showed symptoms of water intoxication at the time of diagnosis. Of the eight patients who had plasma ADH measured at diagnosis, seven had elevated values (mean 52.0, range 16.1 - greater than 250 pg/ml). Intensive combination chemotherapy produced objective tumor responses in all patients, and syndrome of inappropriate ADH secretion (SIADH) resolved in 16 of 17 evaluable patients within three weeks of initiation of treatment. ADH values after therapy were normal, and all patients maintained a normal serum sodium during the period of tumor remission in spite of unrestricted fluid intake. All 17 evaluable patients have developed progressive cancer, but only 10 have manifested recurrent SIADH. Patient survival was similar to the overall population of small cell carcinoma patients without SIADH. The indirect methods of treatment for SIADH (fluid restriction, demeclocycline, lithium, urea) are frequently of transient value while awaiting a response to chemotherapy or in patients with resistant tumors. However, the initial treatment of choice for SIADH associated with small cell carcinoma of the lung is combination chemotherapy. PMID- 6295594 TI - Immunoglobulin class of antibody to herpes simplex virus in patients with oral cancer. AB - An enzyme-linked immunosorbent assay was used to measure antibody of the IgG, IgA, and IgM classes against Herpes simplex virus type 1 (HSV-1) in human sera. Patients with untreated oral cancer had higher levels of IgM antibody to HSV-1 than did either patients with acute or recurrent herpetic infections or age matched control subjects. Levels of IgM antibody to cytomegalovirus and total serum IgM concentrations were similar in all groups. Patients who had been treated successfully for oral cancer more than one year earlier did not have higher levels of IgM antibody to HSV-1. The results are consistent with the hypothesis that cancer of the mouth is associated with expression of HSV-1 antigens that stimulate IgM rather than IgG antibody responses. PMID- 6295595 TI - Complete recovery of histiocytic medullary reticulosis-like syndrome in a child with acute lymphoblastic leukemia. AB - A 6-year-old boy with acute lymphoblastic leukemia (ALL) developed a haemophagocytic syndrome resembling histiocytic medullary reticulosis (HMR) but made a complete recovery on supportive treatment. This was subsequently found to have been associated with a parainfluenzal infection. It is suggested that HMR in immunocompromised hosts may represent a reactive process to an opportunistic viral infection and that the use of chemotherapy in these patients may be deleterious. PMID- 6295596 TI - Induction of remission in hepatocellular carcinoma. A comparison of VP 16 with adriamycin. AB - Following a pilot study of VP-16.213 (180 mg/m2 on 3 consecutive days at 2 weekly intervals) in the treatment of patients with hepatocellular carcinoma, the efficacy of the drug was compared with that of adriamycin in another 35 patients in a randomized crossover trial. Each drug gave a similar response rate (18 and 28%, respectively) but the duration of response was significantly longer in those receiving Adriamycin. Some patients who had not responded to treatment with Adriamycin had worthwhile remission with VP 16. PMID- 6295597 TI - Application of Epstein-Barr virus (EBV) serology to the diagnosis of North American nasopharyngeal carcinoma. AB - A cooperative study was established among a number of institutions in the USA to determine the clinical value of Epstein-Barr virus (EBV) serology for the diagnosis of different histopathological types of North American nasopharyngeal carcinoma (NPC) including occult primary tumors. One hundred-twenty-four patients with confirmed NPC have now been entered into the study. For each patient, anti EBV antibody titers were determined at diagnosis and related to the histopathology as classified according to the World Health Organization (WHO). The results suggest that certain anti-EBV antibodies are of potential value for the diagnosis of undifferentiated types of NPC but not for the well differentiated cancer. The IgA anti-VCA antibody response is the most specific for this disease and of the greatest diagnostic value when used alone or in combination with the anti-EA test. These tests have also been used successfully for the detection of occult NPC. These results indicate that these tests can be useful aids to the clinician for the diagnosis of certain histopathologic types of this disease. PMID- 6295598 TI - Tissue binding of lectins in disorders of the breast. AB - Twenty breast lesions including seven scirrhous ductal carcinomas, one infiltrating lobular carcinoma, one colloid carcinoma, four fibroadenomas, and seven cases of fibrocystic disease were analyzed by fluorescence microscopy for the presence and distribution of lectin-binding carbohydrates. Paraffin-embedded tissue sections were tested with wheat germ agglutinin (WGA), Ricin communis agglutinin I (RCA I), peanut agglutinin (PNA), Soybean agglutinin (SBA), Dolichos biflorus agglutinin (DBA), Ulex europaeus agglutinin I (UEA I), and concanavalin A (Con A). Brightest and most consistent staining regardless of the nature of the breast lesion was obtained with WGA followed in approximate order of staining intensity by RCA, PNA, SBA/DBA and Con A. UEA I stained many of the benign breast lesions but no malignant lesions. Lectin binding carbohydrate in benign lesions was localized mainly along the apices of mammary epithelial cells but there was considerable variation in staining patterns among malignant tumors. The fluorescence microscopic arrangement of lectin binding carbohydrate appears distinct for each malignant neoplasm of breast but is more consistent in benign conditions. PMID- 6295599 TI - Growth patterns and prognosis in early gastric carcinoma. Superficially spreading and penetrating growth types. AB - One hundred and sixty-seven cases of early gastric cancer were analyzed in terms of growth patterns and were then classified into the Small mucosal type, the superficially spreading (Super) type and the penetrating growth (Pen) type, the latter further subdivided into the Pen A type growing expansively and the Pen B type deepening infiltratively. The Small mucosal, Super, and Pen B types are characterized by a tendency toward dominant depressed lesions, a low incidence of vessel invasion and lymph node metastasis, and a good prognosis after surgery (about 90% of the patients survived for ten years). In contrast, the Pen A type is characterized by dominant elevated lesions usually comprised of well differentiated carcinoma, a relatively high incidence of vessel invasion and lymph node metastasis and a poor prognosis after surgery (64.8% five-year survival), due to early recurrence in the form of liver metastasis. Some attention is given to the related surgical approaches. PMID- 6295600 TI - Metastatic carcinoma. The lung as the site for the clinically undiagnosed primary. AB - Of 387 patients who died with lung cancer, 28 cases were reviewed (7.2%) which were clinically undiagnosed. The male:female ratio was 3.6 and mean age was 64 years in the males, 47 years in the females. The most frequent presenting symptoms were neurologic. Prior to death, 21 patients had known or suspected metastatic disease (biopsy-proven in 12), while a malignant diagnosis was not considered in seven patients. Mean survival was 3.5 months. Despite a mean tumor size of 2.8 cm, most of the chest x-rays were not diagnostic even in retrospect. At autopsy, 65% of the tumors were adenocarcinomas (compared to 32% in the other 359 patients); 53% of these showed vascular and lymphatic invasion around the primary tumor, explaining their wide dissemination. In patients with small cell carcinomas (25% of the cases reviewed) or with solitary metastases (14% of the cases reviewed) therapeutic intervention could possibly have been beneficial. PMID- 6295601 TI - VP-16 plus cyclophosphamide in the treatment of advanced lung cancer. AB - Ninety previously untreated patients with histologically documented lung cancer were treated with VP-16 and cyclophosphamide either alone (protocol I) or with methotrexate (protocol II) or Adriamycin (protocol III), with 30 patients in each protocol. The rates of objective response were 57,37, and 27%, respectively, protocol I being significantly better than protocol III (P less than 0.05). Protocol I was significantly less toxic than protocols II(P less than 0.01) and III (P less than 0.001). The overall rate of objective responses was 66% in small cell (SCC) and 22% in non-small cell carcinoma (nSCC). Median survival was 37 weeks in SCC and 21 weeks in nSCC. Median survival of responders both in SCC and in nSCC was significantly longer than in nonresponders. We conclude that VP-16 plus cyclophosphamide is a well tolerated regimen with positive effect in advanced lung cancer. The association of methotrexate or Adriamycin didn't offer any improvement over the basic combination in this study. PMID- 6295602 TI - Intravascular, bronchiolar, and alveolar tumor of the lung (IVBAT). An analysis of twenty cases of a peculiar sclerosing endothelial tumor. AB - Twenty cases of an unusual tumor of the lung are described. This tumor usually presents with multiple small, slowly growing pulmonary nodules. Many cases are detected incidentally. Eighty percent are women, and 50% are less than 40 years of age. Survival with tumor can be quite long. However, one half the patients have died, usually of progressive pulmonary insufficiency. This is a peculiar sclerosing tumor of endothelial cell origin. PMID- 6295603 TI - Primary bronchial tumors in childhood. A clinicopathologic study of six cases. AB - A primary bronchial tumor was diagnosed in six children younger than age 12 years. These cases include four bronchial carcinoids, a low-grade mucoepidermoid carcinoma, and a granular cell tumor ("myoblastoma"). Lobar or segmental resection was performed in each case and remains the treatment of choice. The most common bronchial tumors in childhood are carcinoids with 17 other evaluable cases in the English Literature. This series includes the youngest child to be reported with bilateral choroidal metastases leading to blindness; in addition, there were clinical manifestations of gigantism and acromegaly possibly related to ectopic hormone production. Low-grade mucoepidermoid carcinomas rank second in frequency to carcinoids and have an excellent prognosis. Judging from the childhood bronchial tumors reported to date, proved examples of bronchogenic carcinoma are vanishingly rare. PMID- 6295604 TI - Epstein-Barr virus, fatal infectious mononucleosis, and Hodgkin's disease in siblings. AB - Epstein-Barr virus (EBV) infection in a family resulted in a fatal disseminated heterophil negative infectious mononucleosis syndrome in a nine-year-old girl. This was followed closely by a similar disease process in her six-year-old brother which evolved over a one-year period into Stage IIIB Hodgkin's disease. Finally, three years after the index EBV case in the daughter, the mother was diagnosed with a non-Burkitt's-type undifferentiated lymphoma that proved rapidly fatal. The EBV involvement in the sister and brother was well documented serologically and virologically. The pathologic diagnosis was established and confirmed by more than one pathologist. There was no obvious evidence for either a specific or general immune defect in any of the family members tested. The progression of the six-year-old boy's EBV infection from a benign, yet disseminated disease process into a histopathologically confirmed case of Hodgkin's disease offers a strong suggestion that this virus was not behaving solely as a passenger. Especially relevant is the fact that the boy never fully recovered from his EBV infection and essentially became persistently infected with the virus as evidenced by his EBV-EA serology and virology results. PMID- 6295605 TI - Breast carcinoma after cancer therapy in childhood. AB - Among 910 survivors of childhood cancer, four developed infiltrating carcinoma of the breast and another had noninfiltrating breast tumor. Expected frequency was 0.3 cases of breast cancer in the series. The affected women developed breast carcinoma at ages 20, 25 and 38 years, and the men at ages 38 and 39 years, respectively. Each patient had received orthovoltage chest irradiation for treatment of Wilms' tumor or bone sarcoma between seven and 34 years previously, and estimated radiation dose to the breast exceeded 300 rad in each instance. Four patients also received diverse forms of chemotherapy. Survivors of childhood cancer have increased risk of developing breast cancer and should undergo periodic screening, particularly after breast tissue had been irradiated. Individualized radiotherapy planning can help exclude the breasts from treatment fields for some thoracic neoplasms. PMID- 6295606 TI - Intracranial metastases in small cell carcinoma of the lung. Prognostic aspects. AB - Two hundred-twelve consecutive patients with small cell carcinoma of the lung were studied in order to correlate the risk of developing intracranial metastases to the initial stage of the disease (locoregional versus extensive) and to evaluate the prognostic significance of developing intracranial dissemination of the disease. Clinically detected intracranial metastases were observed in four percent at the time of primary diagnosis, and an additional 18 percent developed metastases during treatment. As regards clinically observed metastases during treatment, no difference was found between the two initial staging groups. Intracranial metastases without clinical evidence of progressive disease elsewhere were demonstrated in 10 out of 205 patients (5%). The median survival time after clinical presentation of intracranial metastases was 85 days for patients with locoregional disease versus 60 days for patients with extensive disease. A significantly shorter survival time was observed for patients with intracranial metastases at 0, 100, 200 and 300 days after start of treatment compared to patients still alive without metastases at those times. Brain autopsy was performed in 82 patients and was positive in 42 (51%). No statistical difference in the frequency of brain metastases was demonstrated when compared to the initial stage of the disease. No difference was observed between the two initial staging groups of patients with regard to risk of developing brain metastases. Autopsy substantiated that there was no difference between patients with and without brain metastases as regards survival. However, clinical intracranial metastases were followed by a short survival time, and only a small fraction of the patients developed clinically isolated intracranial relapse. PMID- 6295607 TI - Malignant fibrous histiocytoma in the lung masquerading as recurrent pulmonary thromboembolism. PMID- 6295608 TI - Hepatocellular carcinoma with situs inversus. PMID- 6295609 TI - An analysis of the role of radiotherapy alone and in combination with chemotherapy and surgery in the management of advanced breast carcinoma. AB - One hundred-eight consecutive patients with Stage III breast carcinoma received radiotherapy as the primary method of treatment. Of these, 53 had prior chemotherapy and 47 had a postirradiation mastectomy as planned adjunct modalities. The overall actuarial survival probability is 40 +/- 6% at five years and 14 +/- 6% at ten years. The corresponding probabilities of freedom from metastases are 27 +/- 5% and 14 +/- 5%, while those of local control are 58 +/- 8% and 52 +/- 8% respectively. The patients receiving chemotherapy show both increased five and ten year probabilities of metastatic control of 38 +/- 7% and 22 +/- 8% and also a five and ten year local control rate of 73 +/- 8%. The patients with additional mastectomy exhibit an improved five year local control rate of 80 +/- 8% but the probability of remaining free from metastatic disease is unaltered being 31 +/- 7% at five and 12 +/- 7% at ten years. The 23 patients receiving chemotherapy prior to radiotherapy, without a subsequent mastectomy have the highest metastatic control of 41 +/- 12% at five years. In this group, eight patients treated with time-dose factors (TDFs) of over 80 have had only one local failure with a preliminary five year local control probability of 80 +/- 18% and metastases-free rate of 58 +/- 19%. Based on these results recommendations are made for minimum tumor doses of 6000 rad and adjuvant chemotherapy. This should be delivered before radiotherapy, since with this sequence of treatment both local control and survival are improved. PMID- 6295610 TI - Adenoid cystic carcinoma of the salivary gland. Sustained complete response to chemotherapy. AB - A patient developed widely metastatic adenoid cystic carcinoma of the parotid gland after several local recurrences. A complete regression of pleural, pulmonary parenchymal, cutaneous, and bone metastases was seen following therapy with 5-fluorouracil, Adriamycin, and mitomycin C. This is the second complete response to chemotherapy reported for a patient with metastatic adenoid cystic carcinoma of salivary gland origin. Both complete responses and several partial responses have followed treatment with Adriamycin, suggesting that this drug is active in the disease. PMID- 6295611 TI - Murine mammary tumor virus related antigen in human male mammary carcinoma. AB - An antigen immunologically related to mouse mammary tumor virus (MuMTV) and the major envelope glycoprotein, gp52 of MuMTV, was identified in tissue sections of human male and female mammary carcinomas using the peroxidase-antiperoxidase technique. The specificity of the reaction was established by absorption studies. Positive reactions with the gp52 antiserum were seen in mouse and human mammary carcinomas, but not in normal mammary tissues, mammary tissues with benign diseases and in other primary malignant neoplasms. Almost all (32/36, 89%) male mammary carcinomas were positive for the gp52 related antigen. A lesser proportion of tumors among female patients (14/50, 28%) were positive. The gp52 positive tumors were significantly larger than the gp52 negative tumors in female patients (P less than 0.05). Gp52 positive tumors were also larger than gp52 negative tumors in male patients, but the difference was not statistically significant. Gp52 reactivity was also detected in metastatic mammary carcinoma in axillary lymph nodes of male and female patients. The presence of gp52 related antigen was not apparently related to tumor grade or lymphocytic infiltrate in the primary tumor. The data do not permit a firm conclusion regarding nodal status in men; no correlation of gp52 activity and nodal status in women was evident. These results indicate that mammary carcinomas in men as well as in women have an antigen related immunologically to MuMTV gp52. Other than tumor size, the antigen seems to be unrelated to major prognostic factors. The significance of the antigen with respect to etiologic features and prognosis in breast cancer remains to be determined. PMID- 6295612 TI - The natural history of lung cancer with pericardial metastases. AB - Sixty-five lung cancer patients with pericardial metastases were reviewed. The diagnosis was established antemortem in 30 patients. Median survival of the treated group for pericardial metastasis from the date of primary diagnosis of lung cancer was 22.5 weeks. Patients who did not receive treatment had a median survival of 1.75 weeks (P 0.01). Median survival, in the treated group, from the date of pericardial metastases was 21 weeks. Prolonged survival was seen in those who had involvement of pericardium only as the site of first metastases. Median survival of this group was 32.5 weeks as opposed to those who had pericardial as well as other metastases whose survival was 17.25 weeks (P 0.03). Radiation therapy appeared to be the best modality of treatment in this group of patients. PMID- 6295613 TI - Pedunculated hepatocellular carcinoma. Report of three cases and review of literature. AB - Pedunculated hepatocellular carcinoma (HC) is rarely found in the United States and Europe. There have been several cases in Japan. The diagnosis of pedunculated HC is said to be very difficult. This article describes three cases of pedunculated HC which were diagnosed preoperatively and were resected successfully. Celiac angiography and CT scan were useful diagnostic procedures. World literature is reviewed, and 15 documented cases of pedunculated HC have been found, including the three cases presented. It may be assumed that pedunculated HC may arise from the accessory lobe of the liver or ectopic liver tissue. PMID- 6295614 TI - Therapy of malignant APUD cell tumors. Effectiveness of DTIC. AB - Eleven patients with malignant APUD tumors, five islet cell carcinomas, five carcinoid tumors and one medullary carcinoma of the thyroid were treated with DTIC. Nine of 11 patients benefitted from treatment. A literature review revealed that other APUD tumors responded when treated with DTIC. DTIC is a useful agent for treatment of malignant APUDomas, and may be the drug of choice for islet cell carcinoma of the pancreas. PMID- 6295615 TI - Chemotherapy of malignant fibrous histiocytoma of bone. A report of five cases. AB - Five patients with evaluable malignant fibrous histiocytoma (MFH) of bone (three with primary tumor and two with primary tumor and metastatic disease) were treated with preoperative chemotherapy including high dose methotrexate (HDMTX) with citrovorum factor rescue (CFR) as is used for patients with osteogenic sarcoma. All five patients demonstrated a clinical response to chemotherapy. Three of four patients who underwent surgery had complete responses and one patient had greater than 90% tumor necrosis as documented by histologic examination of the resected primary tumor. All four patients who underwent surgery following preoperative chemotherapy are surviving free of disease from one to six years from the start of treatment; chemotherapy was discontinued after six to 11 months in these patients. The median disease-free survival time is 31.5 months. This study demonstrates the effectiveness of chemotherapy in MFH of bone, and in particular the effectiveness of HDMTX with CFR which caused measurable responses in all patients while receiving this therapy as a single agent. PMID- 6295616 TI - Hepatitis B surface antigen positive hepatocellular carcinoma in children. Report of a case and review of the literature. AB - This report describes a case of adult type hepatocellular carcinoma (HCC) of a ten-year-old boy. The liver showed no cirrhosis but hepatitis B surface antigen (HBs-antigen) was demonstrated in the non-neoplastic liver cells. Serological examination of his family revealed positive HBs-antigen of the patient's mother suggesting familial transmission to the patient in his early life. Histologic demonstration of HBs-antigen was performed on 36 adult HCC cases and 14 childhood cases including three HCC and eleven hepatoblastoma. HBs-antigen was demonstrated in 12 adult cases, one of which showed no cirrhosis. All childhood cases except for the one presented here showed no HBs-antigen. Review of the literature disclosed four other cases of HBs-antigen positive HCC in children. The present case is unique in that the non-neoplastic portion of the liver showed neither hepatitic nor cirrhotic manifestations. These HBs-antigen positive HCC in childhood may imply the significant relationship between HB virus and HCC not only in adult but also in childhood. PMID- 6295617 TI - Pathology of hepatocellular carcinoma in Japan. 232 Consecutive cases autopsied in ten years. AB - The pathologic findings of 232 consecutive cases of hepatocellular carcinoma (HCC) autopsied during the past ten years at Kurume, Japan, were analyzed from the point of view of global epidemiology, in relation to clinical feature, and in regard to incidence, age, sex, etiologic factors, size of liver, changes in noncancer parenchyma, gross type of tumor, extrahepatic metastases, intravascular and intraductal growths, cancer cell histology, hepatitis B surface antigen (HBsAg) in hepatocytes and cancer cells, liver cell dysplasia, and frequency and clinicopathologic characteristics of minute HCC. Furthermore, postmortem hepatic arteriography and portography were done in 152 livers for comparison with gross anatomy and celiac angiograms. It was found that: (1) epidemiologically, HCC in Japan is distinct from that in the West that it is frequently encapsulated, livers are generally small because of frequent and advanced cirrhosis and small cancer, minute HCC, is not uncommon at autopsy, cirrhosis most commonly associated is the one with thin stroma and medium size nodules, and micronodular cirrhosis is very rare despite frequent alcohol abuse; (2) HCC is increasing in incidence; (3) HBsAg is frequently found in parenchyma; (4) liver cell dysplasia is indirectly related to HBsAg with no evidence for premalignancy; (5) the lung is the most frequent site of metastasis but peritoneal dissemination is unusual; (6) intraportal tumor growth is very common and the hepatic vein is less frequently affected; (7) growth in the major bile duct is frequently associated with intraportal growth and clinically presents as obstructive jaundice; and (8) tumor is supplied solely by arteries and celiac arteriograms are closely correlated with gross pathologic findings. PMID- 6295618 TI - Renal sarcomas of childhood. A clinicopathologic and ultrastructural study. AB - Of 230 consecutive primary renal tumors of childhood collected over a 26-year period, 19 (8.2%) were sarcomas. The histologic criteria devised by the National Wilms' Tumor Study for diagnosis of "sarcomatous variants" of Wilms' tumor could be applied to all but three cases, namely, a rhabdomyosarcoma, a spindle-celled tumor, possibly leiomyosarcoma, and an unclassified primitive tumor. The most prevalent subtype was the bone-metastasizing, "clear cell" sarcoma. Correlative histologic and ultrastructural study supports the concept of subsets of tumors within the category now called "sarcomatous Wilms' tumor." Although this designation has the practical value of alerting the oncologist to the need for aggressive therapy, an origin in nephrogenic mesoderm cannot be cogently proposed for all of these tumors. This study also shows that the histologic features of the different subtypes of renal sarcomas may overlap. Hence, electron microscopy is indispensable for appropriate subclassification of these tumors. PMID- 6295619 TI - The effect of CNS metastases on the survival of patients with small cell cancer of the lung. AB - The records of 227 patients with small cell lung cancer (SMCLC) treated between January 1974 and July 1978 in a series of randomized trials were reviewed to determine the influence of central nervous system (CNS) metastases on survival. Sixteen patients were excluded because of single agent chemotherapy (11), lack of CNS irradiation despite proven metastases (2), prior chemotherapy (2), and concomitant metastatic second primary (1). Of 211 evaluable patients, 100 presented with limited disease and 111 with extensive disease, 25 of whom had CNS metastases at presentation, 21 ("CNS-limited") as the only site of metastases. Treatment of limited patients consisted of chemotherapy and thoracic radiation, while chemotherapy alone was used for extensive patients. No prophylactic brain irradiation was used, but CNS radiation was given to almost all patients when CNS metastases developed. Median survivals were: limited, 13.8 months; CNS-limited, 15.1 months; and extensive 8.6 months (P less than 0.0001). There was no significant difference in the survival experience of limited and CNS-limited patients, although none of the CNS-limited patients experienced long-term remissions. Thirty-five of the limited patients and 21 of the extensive patients subsequently developed CNS metastases. Their median survivals following CNS metastases were 3.7 months and 1.6 months, respectively. In conclusion, CNS metastases as the sole site of metastatic disease at diagnosis of SMCLC is not necessarily a bad prognostic sign, while the subsequent development of CNS metastases may be. PMID- 6295621 TI - Results of retroperitoneal lymph node dissection and postoperative adjuvant chemotherapy with dactinomycin in the treatment of retroperitoneal metastases of nonseminomatous testicular germ cell tumors. AB - This article discusses 103 patients with a nonseminomatous testicular germ cell tumor. Treatment of 87 patients in clinical Stages I, II-A, and II-B consisted of bilateral retroperitoneal lymph node dissection. In Stage I (54 patients), no further treatment was given. The three-year survival was 96%, while recurrence free survival was 98%. Of 33 patients in Stages II-A and II-B, 26 received adjuvant chemotherapy with dactinomycin, and ten of these were given radiotherapy (peritoneum/mediastinum) as well. The three-year survival was 88% and recurrence free survival was 85%. Seven patients in Stages II-A and II-B received no adjuvant chemotherapy; four of them died. Treatment of 14 patients in Stage II-C primarily consisted of dactinomycin therapy, subsequently combined with surgery and radiotherapy. The three-year survival was 28.5%. These findings raise the question whether adjuvant single-agent chemotherapy with dactinomycin for patients with a nonseminomatous testicular germ cell tumor should not be re evaluated. PMID- 6295620 TI - High-dose methotrexate in small cell lung cancer. Lack of efficacy in preventing CNS relapse. AB - Few studies have incorporated high-dose methotrexate (MTX) with leucovorin rescue in the treatment of small cell lung cancer (SCLC). Potentially therapeutic levels of MTX can be achieved in the central nervous system (CNS) by systemic administration of high doses of this drug. Utilizing a combination chemotherapy program of Adriamycin, vincristine, cyclophosphamide, and methotrexate, 31 patients were sequentially assigned to receive either low-dose MTX (40 mg/m2), or high-dose MTX (500 mg/m2) with leucovorin rescue. Radiation therapy to the primary site was also administered. At these dosage levels there were no statistically significant differences in response rate or survival between the two groups. High-dose MTX did not prevent the appearance of CNS disease; there being 2/15 and 3/15 CNS relapses in the HD MTX and LD MTX treated groups, respectively. The occurrence of CNS disease did not significantly affect overall survival as compared to patients not similarly affected. PMID- 6295622 TI - Glucagonomas. Ultrastructure and immunocytochemistry. AB - Pancreatic tumors harboring glucagon immunoreactive cells were found in four patients with diabetes mellitus. Alpha-cell (glucagon) granules were present in three tumors; pancreatic polypeptide (PP) immunoreactive cells were detected in two. In two patients the tumors were malignant and one of these had the glucagonoma syndrome; the other was a member of a family with MEN-type I syndrome. These cases illustrate three clinical subtypes of glucagonoma. PMID- 6295623 TI - Effect of tunicamycin on receptors for tumor promoters. AB - A progressive decline in the specific binding of [20-3H]phorbol 12,13-dibutyrate ([3H]PDBu) and glycoprotein synthesis was observed following treatment of primary mouse epidermal cells with tunicamycin, a specific inhibitor of dolichol-mediated glycosylation. Following 18 h of treatment, the specific binding of [3H]PDBu was reduced to 33-56% of the control value. The total protein synthesis determined by leucine incorporation into acid-insoluble material was not altered by this antibiotic drug. These results suggest that the receptor for phorbol diesters is, or is functionally linked to, a glycoprotein on the cell surface. PMID- 6295624 TI - Comparative tumor initiating activity on mouse skin of 6-nitrobenzo[a]pyrene, 6 nitrochrysene, 3-nitroperylene, 1-nitropyrene and their parent hydrocarbons. AB - 6-Nitrobenzo[a]pyrene, 6-nitrochrysene, 3-nitroperylene, 1-nitropyrene, and the corresponding parent hydrocarbons were tested for tumor initiating activity on mouse skin with promotion by tetradecanoylphorbol acetate. The initiating doses of 6-nitrobenzo[a]pyrene and benzo[a]pyrene were 0.05 mg each; for all other compounds the initiating doses were 1.0 mg. 6-Nitrochrysene induced tumors in 60% of the mice (2.1 tumors per mouse), but was significantly less tumorigenic than chrysene. 3-Nitroperylene induced tumors in 42% of the mice (0.5 tumors per mouse) and was significantly more active than perylene. Neither 1-nitropyrene nor 6-nitrobenzo[a]pyrene exhibited significant tumorigenic activity in the concentrations tested. PMID- 6295626 TI - Formation of a new DNA replication intermediate in novobiocin-treated human melanoma cells. AB - The effect of novobiocin on the formation of DNA replication intermediates in human melanoma cells was investigated. This drug reduces DNA synthesis in all probability by interfering with the DNA gyrases (enzymes that are involved in the supercoiling of the DNA). The single-stranded DNA replication intermediates were released from the parental DNA by lysing the cells in dilute alkali and were then fractionated by agarose gel electrophoresis. In the untreated cells one can detect replication intermediates ranging in size from Okazaki-fragments up to 10 kb. However, in novobiocin-treated cells, apart from the already known intermediates, formation of a new replication intermediate with the size of about 20 kb was also detected. The 20-kb DNA shows a similar discrete appearance in the agarose gel as the 10-kb DNA intermediate. PMID- 6295625 TI - Promotional effect of different phorbol esters on morphological transformation of hamster embryo cells. AB - Hamster embryo cells were sequentially exposed to benzo[a]pyrene (BaP) (3 days) and different phorbol esters (4 days). Compounds which have been shown to act as tumor promoters in mouse skin carcinogenesis, showed a promotion-like effect on the formation of morphologically transformed colonies, while non-promoting analogs did not enhance the transformation frequency. In experiments where the exposure sequence was reversed, i.e. 12-O-tetradecanoyl phorbol-13-acetate (TPA) in the first exposure period and BaP in the second period, no significant enhancement of the transformation frequency was observed. The exposure to a low concentration of BaP in the second exposure period following a higher concentration in the first period, resulted in a strong enhancement of the transformation frequency suggesting that BaP also acts as a promoter. PMID- 6295627 TI - Role of plasma fibronectin in the morphological transformation of hamster embryo cells. AB - Replacement of fetal bovine serum (FBS) by newborn serum in the hamster embryo cell transformation assay blocks the induction of morphologically transformed colonies by chemical carcinogens. Moreover, the addition of newborn serum strongly inhibits the formation of morphologically transformed colonies in the standard assay using 20% FBS. The present experiments show that the inhibitory factor is heat labile and not dialyzable, but is removed when the newborn serum is submitted to gelatin affinity chromatography. The inhibitory factor is recovered in the eluted gelatin bound material which gives essentially one band in SDS electrophoresis corresponding to that of plasma fibronectin. In separate experiments fibronectin from bovine plasma is shown to inhibit cell transformation in a similar way. The experiments suggest that fibronectin plays a central role in the mechanism of morphological cell transformation. PMID- 6295628 TI - An overview of slow channel blocking drugs: pharmacological basis for therapeutic applications. AB - The use of voltage clamps in cardiac muscle has permitted the separation of two inward depolarizing membrane currents. The first is the fast sodium current, with rapid kinetics of activation and inactivation, which is blocked by local anesthetics. The second is the slow current carried largely by calcium ions, having slow kinetics with a long time constant of inactivation. Agents which selectively block this current are called Ca antagonists. Such agents are heterogeneous structurally; they also block calcium currents in smooth muscle and therefore produce coronary as well as peripheral vasodilatation. The most significant agents are verapamil, gallopamil, nifedipine, diltiazem and tiapamil. They have a varying spectrum of pharmacological effects with respect to changes they produce on heart rate, afterload, preload, contractility, coronary flow and on the AV node. A similar spectrum of therapeutic effects is apparent in different clinical disorders, in particular vasospastic angina, stable and unstable angina, cardiac arrhythmias, hypertension, hypertrophic cardiomyopathy and possibly other disorders. Indeed, the full range of clinical disorders which may respond to calcium antagonists is still not fully delineated and may well be at least as extensive as that for beta-adrenoceptor blocking drugs. The advent of calcium antagonists in the last decade represents one of the most significant advances in cardiovascular therapeutics. PMID- 6295629 TI - Influence of HClO4 strength and etching time on rate of etching and surface roughness of human enamel. PMID- 6295630 TI - [The Epstein-Barr virus and tonsilar carcinoma in clinical practice]. PMID- 6295632 TI - An organ culture study on the site of determination of ACTH and LH cells in the rat adenohypophysis. PMID- 6295631 TI - Effects of brain and mesenchyme upon the cytogenesis of rat adenohypophysis in vitro. I. Differentiation of adrenocorticotropes. AB - This study investigates the role of the developing diencephalic floor or mesenchymal tissue in the differentiation of ACTH-producing cells. The adenohypophysial primordia of fetal rats on days 12.5 and 13.5 of gestation were treated with collagenase; some primordia were allowed to retain an association with the brain and mesenchyme, but in others the brain and/or mesenchyme were removed. These different combinations of tissues were cultured and examined by immunohistochemical techniques using antisera against pACTH and synthetic alpha MSH. Removal of mesenchyme alone had little effect on the development of ACTH cells as compared to primordia maintained with brain and mesenchyme. In contrast, removal of the brain with or without mesenchyme on day 12.5 resulted in a marked decrease of ACTH cells accompanied by a mal-growth of adenohypophysial tissue. Such changes were slight when the brain was separated from day 13.5 primordia. Immunoreactive alpha-MSH cells were sparse or absent in all cases. These results suggest that in fetal rats the developing diencephalic floor is essential for differentiation of ACTH cells before day 13.5 of gestation whereas mesenchyme has no apparent effect. PMID- 6295633 TI - Immunohistochemical demonstration of a CCK-like peptide in the nervous system of a marine annelid worm, Nereis diversicolor O.F. Muller. AB - Perikarya and nerve fibers containing a substance immunologically related to CCK 8 were detected in the nervous system of Nereis, a marine annelid worm. The most noteworthy immunostaining was seen in cell bodies, localized at the periphery of the brain, within nuclei 5, 6, 7, 9, 10, 13, 14, 15, 17, 20, 23-24. Immunoreactive fibers were also found in the neuropile without any particular grouping. Numerous other "positive" perikarya occur in the medioventral portion of the ventral nerve cord, and in the ventral and dorsal parts of the suboesophageal ganglion. In addition to the cell bodies in the cerebral external layer, immunoreactive axons were abundantly observed in the connectives between the ganglia. Moreover, our results demonstrate CCK-like staining in neurons showing variations in size and shape, and in affinity for paraldehyde fuchsin. The present results support the hypothesis that this peptide may exert a role as neurotransmitter or neuromodulator in annelids. PMID- 6295635 TI - Headache syndromes suggested by statistical analysis of headache symptoms. AB - Statistical study of the correlation between 49 variables concerning characteristics of headache and of headache patients was carried out on 1,198 sequential patients complaining of severe or disabling headaches and attending a headache clinic. Factor analysis was the statistical method employed, 17 factors accounting for 64% of total variation emerged. The most prominent factor contained descriptors of neurological dysfunction associated with headache. Another factor contained variables of nausea and vomiting, only associated with inability to work. Other factors, contained separately, the following variables: (a) Time of headache, (b) time of week of headache, (c) unilateral nature of headache, (d) visual phenomena associated with headache. Relation of these findings to headache taxonomy is discussed. PMID- 6295634 TI - Exocytosis in isolated gastric glands induced by secretagogues and hyperosmolarity. AB - Gastric glands were isolated from rabbit gastric mucosa and incubated in the presence of secretin, cholecystokinin octapeptide (CCK-8), and in hyperosmolar medium. These agents all elicited a compound exocytotic response from the chief cells observed by electron microscopy. Responses to secretagogues yielded significantly higher values than controls at 2.5 min after exposure, and were essentially complete within 30 min. The response to hyperosmolarity was more gradual and continued over a 60 min period. Glands incubated in hyperosmolar medium were still capable of responding to secretagogues, and the combination was additive. The results indicate that both secretagogues, which initiate specific receptor mediated events, and hyperosmolarity, which causes a more generalized change in the cells, result in pepsinogen-granule release by compound exocytosis. PMID- 6295636 TI - Electron microscopic evidence for an asialoglycoprotein receptor on Kupffer cells: localization of lectin-mediated endocytosis. AB - Direct evidence is given for the presence of an N-acetyl-D-galactosamine-specific lectin on the Kupffer cell surface by visualization of ligand binding in electron microscopy. When freshly isolated Kupffer cells are incubated with asialofetuin adsorbed onto colloidal gold particles (ASF-gold), binding and endocytosis of ligand are seen. Recognition of ASF-gold by Kupffer cells is completely abolished in the presence of N-acetyl-D-galactosamine (25 mM) or EGTA (3 mM), but is not significantly reduced by N-acetyl-D-glucosamine or D-mannose (25 mM). ASF particles are endocytosed via the coated pit/vesicle pathway and appear to be transported to the secondary lysosomes by coated vesicles, as shown by the occurrence of coated areas in the secondary lysosome membrane. These observations demonstrate the presence of an asialoglycoprotein receptor on Kupffer cells; therefore, the hepatocyte is not the only cell in the rat liver with D-galactose receptor activity. PMID- 6295637 TI - Recombination in RNA. AB - The aphthovirus genome consists of a single molecule of single-stranded RNA that encodes all the virus-induced proteins. We isolated recombinant aphthoviruses from cells simultaneously infected with temperature-sensitive mutants of two different subtype strains. Analysis of the proteins induced by 16 independently generated recombinants revealed two types of protein pattern, which were consistent with single genetic crossovers on the 5' side and 3' side, respectively, of the central P34-coding region. Recombinants invariably inherited all four coat proteins from the same parent, and novel recombinant proteins were not observed. RNAase T1 fingerprints of virus RNA, prepared from representatives of each recombinant type, confirmed the approximate crossover sites that had been deduced from the inheritance of proteins. These fingerprints provide molecular evidence of recombination at the level of RNA and demonstrate the potential of RNA recombination for producing genetic diversity among picornaviruses. PMID- 6295638 TI - Genetic structure and origin of t haplotypes of mice, analyzed with H-2 cDNA probes. AB - We investigated the genetic organization and evolutionary origin of t chromosomes of mice by examining the restriction fragment patterns of DNA from t haplotypes and normal chromosomes with cDNA probes to H-2 class I genes. On genomic DNA blots, the restriction fragments containing H-2-related sequences were highly variable among different inbred strains of mice, whereas they were very similar among different t haplotypes even when the t haplotypes carried serologically different H-2 haplotypes. These observations suggest that all t haplotypes have a common origin and are not products of independent mutational events. We also mapped the position of several restriction fragments characteristic of t DNA by using a battery of recombinant t haplotypes, defined with respect to their t lethal factors and H-2 haplotypes. We thus show that restriction fragments containing H-2-related sequences map to the left of the H-2 class I genes in t chromosomes, a region in which the tw32 b-lethal factor also maps. The cloning of these fragments can be expected to provide an entry for the structural analysis of t DNA. PMID- 6295639 TI - Ds controlling elements of maize at the shrunken locus are large and dissimilar insertions. AB - A recombinant clone for the wild-type gene of sucrose synthetase has been selected. Subclones of the genomic clone were made and used as probes to construct a restriction map of the wild-type locus. The transcribed region and the direction of transcription in the map were also determined. Four Ds-induced mutations of the locus that were isolated by B. McClintock were analyzed and partially mapped. Two mutations contain insertions of approximately 20 kb of foreign DNA into the transcribed region; two others are at the 5' end of the gene. There are profound differences in the restriction maps of the Ds elements in the mutants analyzed, even though these elements have a common genetic origin and are separated by only a few generations. Comparison of one pair of closely related Ds elements indicates that these differences may be the result of extensive internal rearrangements. Exchange with other elements in the genome remains an alternative possibility. A revertant of one of the Ds-induced mutations has maintained a 21-22 kb insert at the same location in the transcribed region. This insert differs from its predecessor by extensive rearrangement in two thirds of its length. Part, or all, of this rearrangement presumably permits functional expression of the gene. PMID- 6295640 TI - The molecular basis of P-M hybrid dysgenesis: the nature of induced mutations. AB - The molecular nature of mutations arising in dysgenic hybrids between P and M Drosophila melanogaster strains has been investigated. Seven independent mutations at the white locus were examined, and these fell into two classes on the basis of their genetic and structural properties. The five mutations comprising the first class were caused by DNA insertions of 0.5, 0.5, 0.6, 1.2 and 1.4 kb, respectively. The DNA insertions in four of these mutations were examined in detail. Although heterogeneous in size and pattern of restriction enzyme sites, they were homologous in sequence. We refer to members of this sequence family as P elements. Mutations caused by P elements appeared to be stable in the P cytotype, but had reversion rates greater than 10(-3) in the M cytotype. Phenotypic reversion to wild-type was accompanied by excision of the P element. The two mutations comprising the second class were caused by insertion of the 5.0 kb copia element and appeared to be stable in both P and M cytotypes. PMID- 6295641 TI - The molecular basis of P-M hybrid dysgenesis: the role of the P element, a P strain-specific transposon family. AB - We have shown previously that four of five white mutant alleles arising in P-M dysgenic hybrids result from the insertion of strongly homologous DNA sequence elements. We have named these P elements. We report that P elements are present in 30-50 copies per haploid genome in all P strains examined and apparently are missing entirely from all M strains examined, with one exception. Furthermore, members of the P family apparently transpose frequently in P-M dysgenic hybrids; chromosomes descendant from P-M dysgenic hybrids frequently show newly acquired P elements. Finally, the strain-specific breakpoint hotspots for the rearrangement of the pi 2 P X chromosome occurring in P-M dysgenic hybrids are apparently sites of residence of P elements. These observations strongly support the P factor hypothesis for the mechanistic basis of P-M hybrid dysgenesis. PMID- 6295642 TI - Role of transmethylation in the elicitation of an oxidative burst in macrophages. PMID- 6295644 TI - In situ measurement of cAMP related enzymes in the dimorphic fungus Mucor rouxii. PMID- 6295643 TI - Effects of cyclic AMP on in vivo cytotoxic T lymphocytes generation. PMID- 6295645 TI - On the mechanism of TSH-induced formation of follicle-like structures in primary cultures of thyroid cells. PMID- 6295646 TI - Polybrominated biphenyls as aryl hydrocarbon hydroxylase inducers: structure activity correlations. AB - The synthesis of all possible laterally-substituted polybrominated biphenyl (PBB) congeners containing two para bromines is described. Using enzymic, electrophoretic and ligand-binding assays that distinguish between phenobarbitone(PB)- and 3-methylcholanthrene(MC)-type inducers, the synthetic PBBs were evaluated as inducers of liver microsomal drug-metabolizing enzymes in the immature male Wistar rat. 4,4'-Dibromobiphenyl resembled PB in its mode of induction whereas all the meta-brominated derivatives of 4,4'-dibromobiphenyl, namely 3,4,4'-tri, 3,4,4',5-tetra-, 3,3', 4,4'-tetra-, 3,3',4,4',5-penta- and 3,3',4,4',5,5'-hexabromobiphenyl, resembled MC in their mode of induction. The results obtained with 3,4,4'-tribromobiphenyl demonstrate that, in contrast to the polychlorinated biphenyls (PCBs), a single meta halogen substituent is sufficient to abolish the PB-type characteristics of 4,4'-dibromobiphenyl and convert it to a strictly MC-type inducer. PBBs which induce AHH activity must be substituted at both para positions and at one, two, three or four meta positions. Ortho-substitution of PBBs which contain only lateral bromine groups may also give compounds which are aryl hydrocarbon hydroxylase (AHH) inducers. One of the MC-type PBBs, namely 3,3',4,4'-tetrabromobiphenyl, which has been tentatively identified in the commercial PBB mixture, fireMaster BP-6, was at least 50 times more potent as an inducer of AHH activity than the commercial PBB mixture. The induction of AHH by 3,3',4,4'-tetrabromobiphenyl was accompanied by a dose dependent decrease in both thymus and spleen weights. The thymus and/or spleen weights were decreased in rats treated with the other MC-type PBBs which further supports the correlation between the toxicity of the PBBs and their ability to induce AHH. PMID- 6295647 TI - Metabolism and activation of benzo[a]pyrene by mouse and rat skin in short-term organ culture and in vivo. AB - The metabolic activation of BP was examined in mouse and rat skin in vivo and in short-term organ culture. In mouse skin, larger quantities of ether- and water soluble metabolites were formed and more BP became bound covalently to DNA and protein than in rat skin. Qualitative differences in the formation of dihydrodiol metabolites and of BP-deoxyribonucleoside adducts between mouse and rat skin were also observed. Organ culture techniques may not provide a true model of metabolic activation in vivo because it was found that the covalent binding of BP to DNA and protein was reduced in skin maintained in culture despite an accumulation of dihydrodiol and other ether-soluble metabolites. In addition, the proportions of the syn- and anti-isomers of BP-7,8-diol 9,10-oxide involved in the formation of adducts with deoxyguanosine differed between skin treated in organ culture and in vivo. PMID- 6295648 TI - The influence of flow on the metabolism of perfused benzo[a]pyrene by isolated rat lung. AB - In order to investigate the influence of flow and, thus, substrate delivery, on the ability of lung to metabolize foreign compounds, the disappearance of circulating [3H]benzo[a]pyrene ([3H]B[a]P) and the appearance of B[a]P metabolites was monitored in isolated rat lungs from control and 3 methylcholanthrene (3-MC) pretreated rats perfused at low (10 ml/min) and high (45 ml/min) flows. Increasing the flow or 3-MC pretreatment hastened the disappearance of B[a]P from the perfusion medium reservoir and increased the rate of appearance of total metabolites. However, these manipulations affected the appearance of individual metabolites in the medium in different ways. For example, in lungs from control rats the rate of appearance of 7,8-dihydrodiol (7,8-dihydroxy-7,8-dihydro-B[a]P) (7,8-DHD) in the perfusion medium was markedly increased by increasing flow while that of B[a]P-1,6-quinone was minimally affected. In addition, increasing flow increased the concentration of some B[a]P metabolites, such as 4,5-dihydrodiol (4,5-dihydroxy-4,5-dihydro-B[a]P) (4,5-DHD) in the lung tissue of control rats at the end of the perfusion period, but did not effect much change in the concentration of these metabolites in lungs from 3 MC-pretreated rats. The results show that flow, as well as 3-MC pretreatment, may alter the rate at which metabolism of foreign compounds occurs and the temporal profile of metabolites produced by the intact lung. PMID- 6295649 TI - [In vitro effect of Astragalus mongholicus on activity of cyclic nucleotide phosphodiesterases (cAMP-PDE) in liver and spleen cells of mice]. PMID- 6295650 TI - [Inhibitory effect of retinoids on Epstein-Barr virus induction in Raji cells]. PMID- 6295651 TI - [Detection of early NPC from VCA/IgA antibody positive individuals with anticomplement immunoenzymatic method]. PMID- 6295652 TI - [Studies on morphogenesis of herpes simplex virus type II. Cytoplasmic morphogenesis of herpes simplex virus type I under immunoferritin electron microscopy]. PMID- 6295653 TI - Correlative histochemistry of some enzymes of carbohydrate metabolism in preneoplastic and neoplastic lesions in the rat liver. AB - The livers from a total of 51 Sprague-Dawley rats treated with different doses of N-nitrosomorpholine (80-120 mg/l in the drinking water) for up to 14 weeks together with the livers of 28 control animals were histochemically investigated at the cessation of carcinogenic insult and at varying periods thereafter for their glycogen content, basophilia and activities of various enzymes of carbohydrate metabolism: glycogen synthetase, glycogen phosphorylase, glucose-6 phosphatase, glyceraldehyde-3-phosphate dehydrogenase and glucose-6-phosphate dehydrogenase. The enzymatic patterns of normal tissue, preneoplastic and neoplastic lesions were characterized and compared with reference to the morphologically defined stages of tumor development in the liver. The early appearing glycogen storing areas, localized in the peripheral and intermediate lobular regions, did not show significant changes in the histochemically demonstrable activities of the enzymes tested. After cessation of the carcinogen treatment the more pronounced glycogen storage foci which developed within the aforementioned regions of the liver acinus usually showed a reduction in the activities of phosphorylase and glucose-6-phosphatase while the activity of glucose-6-phosphate dehydrogenase, a key enzyme for the pentose phosphate pathway, was increased. The mixed cell foci, neoplastic nodules and tumors which emerged at later stages were characterized by a progressive shift away from glycogen metabolism towards glycolysis and the pentose phosphate pathway, as indicated by an increase in glyceraldehyde-3-phosphate dehydrogenase and glucose 6-phosphate dehydrogenase activities. These changes in enzyme pattern are supportive of a developmental sequence leading from glycogen storage foci through mixed cell foci and neoplastic nodules to hepatocellular carcinomas. PMID- 6295654 TI - Hepatic cyst formation in male rats partially hepatectomized as weanlings during short-term feeding of 2-acetylaminofluorene. AB - Partial hepatectomy (p.h.) of weanling male Sprague-Dawley rats during short-term feeding of 0.03% 2-acetylaminofluorene (AAF) resulted, 12 to 15 months later, in a massive production of hepatic cysts. The weight of the right lateral and caudate lobes plus neoplasms (the left lateral and median lobes were excised at the time of p.h.) accounted for as much as 43% of total body weight. The incidence of hepatic cysts was negligible when animals were not subjected to p.h. during AAF feeding. The feeding of 0.05% phenobarbital (PB) subsequent to AAF + p.h. did not significantly increase the incidence of hepatic cysts. By comparison of the present data with that of other investigators, it can be suggested that susceptibility to cholangioma formation following AAF + p.h. may be determined in part by the developmental stage of the rat and/or its liver at the time of the AAF + p.h. regimen. PMID- 6295655 TI - In vitro formation of methyldiethyldithiocarbamate after the reaction of nitrosoacetoxymethylmethylamine or methylnitrosourea with disulfiram. AB - Analysis by scintillation measurement, mass spectrometry and h.p.l.c. showed that diethyldithiocarbamic acid (DDTC), the main metabolite of disulfiram (DSF), forms methyldithiocarbamate (MeDDTC) when incubated with [14C]nitrosoacetoxymethylmethylamine ([14C]NAMM) or [14C]methylnitrosourea ([14C]MNU) in different media (bacteria, esterases, rat liver 9000 x g supernatant fraction and microsomes). When DSF instead of DDTC was used, MeDDTC was formed only when soluble enzymes were present which are required to split DSF into two DDTC moieties. No physiological methylation of DDTC takes place as was shown by experiments with [3H]MNU. [14C]Methanol, formed by the decay of [14C]NAMM and [14C]MNU was shown to have no alkylating properties. PMID- 6295656 TI - Characterization of benzo[a]pyrene metabolites isolated from muscle, liver, and bile of a juvenile flatfish. AB - Juvenile English sole (Parophrys vetulus) were force fed [3H]benzo[a]pyrene (BaP) and, after 24 h, metabolites were isolated from bile, liver, and muscle. The metabolites were analyzed by t.l.c. and h.p.l.c., and further characterized by u.v., fluorescence, or m.s. analyses. The identities of BaP 7,8-dihydrodiol, BaP 9,10-dihydrodiol, 1-hydroxy BaP, 3-hydroxy BaP, and 9-hydroxy BaP were confirmed. The amounts of BaP 7,8-dihydrodiol plus its conjugates (sulfates and glucuronides) were greater than twice the amounts of BaP 9,10-dihydrodiol plus its conjugates in bile. Liver also contained twice as much BaP 7,8-dihydrodiol as BaP 9,10-dihydrodiol, demonstrating that English sole produced larger amounts of the 7,8-isomer. T.l.c. analysis showed the presence of an unknown metabolite X migrating between BaP 9,10- and 7,8-dihydrodiols. M.s. showed X to be a non vicinal dihydroxy derivative of BaP. Spectral properties of X were similar to those reported for a dihydroxy derivative, presumably the 3,9-dihydroxy BaP, formed from either 3- or 9-hydroxy BaP in the presence of rat liver microsomes. Compared to the dihydrodiols, X was resistant to both glucuronidation and sulfation. The resistance to form water-soluble metabolites may be responsible for the deposition of relatively large amounts (0.11% of the administered dose) of X in muscle; liver contained a much smaller amount (0.01%) of X. Thus, English sole converted BaP into metabolites known to be toxic to mammals, and these metabolites were found in the liver and edible tissue of the fish. PMID- 6295657 TI - A pilot project in molecular cancer epidemiology: determination of benzo[a]pyrene -DNA adducts in animal and human tissues by immunoassays. PMID- 6295659 TI - Effects of the modulation of epoxide hydrolase activity on the binding of benzo[a]pyrene metabolites to DNA in the intact nuclei. PMID- 6295658 TI - 3-Aminobenzamide, an inhibitor of poly ADP-ribose polymerase, decreases the frequency of alkaline labile lesions and increases growth in human fibroblasts exposed to 3-methyl 4-nitroquinoline 1-oxide. AB - Normal human fibroblasts exposed to the mutagen 3-methyl 4-nitroquinoline 1-oxide (3me4NQO) were additionally incubated with or without the inhibitor of poly ADP ribose polymerase, 3 aminobenzamide (3AB) either during or after mutagen treatment. The number of single strand DNA breaks detectable by alkaline sucrose sedimentation at any given time after exposure to this mutagen was reduced by the prior addition of 3AB, regardless of whether this drug was present during or after mutagen exposure. Furthermore, this effect is reversible upon 3AB removal. Finally, cell survival as analysed by cell growth was increased if cells were treated for one hour with 3AB directly following a 30 min exposure to 3-me4NQO. The data presented suggest an additional role for poly ADP ribose polymerase in DNA repair other than that of inhibiting the increase in ligase II, which occurs after exposure to monofunctional alkylating agents. PMID- 6295660 TI - The effects of dietary corn oil on the metabolism and activation of benzo[a]pyrene by the benzo[a]pyrene metabolizing enzymes of the mouse. AB - Male ICR Swiss mice, weighing 16-20 g, were fed ad libitum either a fat-free diet or a diet containing 10% corn oil. After three weeks on these diets, the rates of benzo[a]pyrene (B[a]P) metabolite formation and metabolism to products which covalently bind with macromolecules were compared using hepatic nuclei and microsomal preparations. The maximum activity of B[a]P hydroxylase in microsomes from untreated animals was increased 50% by feeding the corn oil diet, however, B[a]P hydroxylase in microsomes from 3-methylcholanthrene (3-MC)-treated mice was unaffected by diet. In animals treated with phenobarbital, B[a]P metabolism and B[a]P-DNA adduct formation were greater in microsomes from corn oil fed mice compared to those fed the fat-free diet. At a B[a]P concentration of 96 microM, microsomes from corn oil fed untreated mice produced 26% more extractable metabolites and covalent binding to exogenous DNA was increased 46%. At lower substrate concentrations (0.94-15.0 microM B[a]P), B[a]P-DNA and B[a]P-protein binding were 300-400% greater when incubated with microsomes from corn oil fed mice than when incubated with microsomes from mice fed fat-free diet. The apparent Vmax's determined for the formation of each extractable metabolite were increased 1.5-3.0 times by the corn oil diet. Hepatic nuclear B[a]P hydroxylase and nuclear activation of B[a]P to products which covalently bind to DNA in both non-induced and 3-MC-pretreated animals fed the corn oil diet were greater than that observed in animals fed the fat-free diet. B[a]P hydroxylase activities in the lungs of these animals were unaltered by diet. PMID- 6295661 TI - Specificity of rat liver cytochrome P-450 isozymes in the mutagenic activation of benzo[a]pyrene, aromatic amines and aflatoxin B1. AB - The ability of three purified forms of rat liver cytochrome P-450 to metabolically activate benzo[a]pyrene, trans-benzo-[a]pyrene-7,8-dihydrodiol, 2 aminofluorene, aflatoxin B1, dimethylnitrosamine, and a pyrolysis product of tryptophan(3-amino-1-methyl-5H-pyrido(4,3-b)indole) (Trp-P-2) to mutagenic products was examined using Salmonella typhimurium strains TA98 and G46 in a reconstituted monooxygenase system. The isozymes examined were cytochrome P-450 PB (the major phenobarbital inducible form), and the two major 3-MC inducible forms (cytochromes P-448(52) and P-448(55)). Cytochromes P-448(52) and P-448(55) preferentially metabolize 2-aminofluorene and Trp-P-2 to mutagenic products. However, only cytochrome P-448(55) metabolizes benzo[a]pyrene and its 7,8 dihydrodiol derivative to mutagenic products. Both cytochrome P-448(52) and P 448(55) metabolize aflatoxin B1 to mutagenic products at a much faster rate than cytochrome P-450-PB. Dimethylnitrosamine was not activated by any of the isozymes tested. PMID- 6295662 TI - Treatment of chronic orthostatic hypotension with ergotamine. AB - The acute and chronic effects of ergotamine were examined in four patients with chronic orthostatic hypotension. Chronic oral administration of ergotamine tartrate produced significant increases in standing blood pressure and marked clinical improvement, without appreciable recumbent hypertension. The blood pressure increases were not associated with significant changes in plasma norepinephrine or plasma renin activity. No major toxicity was observed at doses of 2-6 mg/day over treatment periods of 3-18 months. Hemodynamic studies on the effects of i.v. ergotamine tartrate (0.25-0.50 mg) revealed that the ergotamine induced increase in blood pressure in the supine position was associated with an increase in total peripheral resistance (from 1616 +/- 165 to 2574 +/- 583 U) without a change in cardiac output. During 45-60 degrees upright tilt, ergotamine increased both total peripheral resistance (1801 +/- 296 to 3262 +/- 1107 U) and cardiac output (2.42 +/- 0.46 to 3.34 +/- 0.54 l/min). Forearm plethysmographic studies revealed decreased forearm blood flow and venous volume and increased vascular resistance with ergotamine. The orthostatic hypotensives had more platelet alpha-receptors (390 +/- 31 receptors/cell) than the control subjects (234 +/- 17 receptors/cell). The increased receptor level was associated with abnormally low circulating levels of norepinephrine and increased pressor responsiveness to infused norepinephrine in three of the four patients. Chronic ergotamine therapy appeared to reduce platelet alpha-receptor number to normal. The results indicate that ergotamine is of value in certain patients with chronic orthostatic hypotension and that the blood pressure effects are related to vasoconstriction in both arterial and venous beds. PMID- 6295663 TI - Hypertension reduces the number of beta-adrenergic receptors in rat brain microvessels. AB - Beta-adrenergic receptor function was measured in cerebral microvessels of spontaneously and DOCA-salt hypertensive rats using 125I iodohydroxybenzylpindolol (IHYP). Both in genetic and in experimental hypertension, a significant decrease in the number of beta-receptor sites was observed, without receptor affinity changes. These results suggest that alterations of central adrenergic regulation of small vessels may participate in the pathogenetic mechanisms leading to the development of the central hypertensive disease. PMID- 6295664 TI - Phosphoribosyl pyrophosphate synthetase and glutathione reductase in erythrocytes from hyperuricaemic and gout patients. AB - The enzymic activities of erythrocyte phosphoribosyl pyrophosphate synthetase (EC 2.7.6.1) and glutathione reductase (EC 1.6.4.2) have been measured in 54 primary gout patients, 35 individuals having hyperuricaemia and 51 healthy controls. Statistical analyses have shown a significant increase (p less than 0.01) in the enzymic activity of erythrocyte PRPP synthetase in both the hyperuricaemic and gout groups compared with the controls. No correlation between activity and age was found in any of the three clinical groups. A significant decrease (p congruent to 0.01) was found in the enzymic activity of red cell glutathione reductase in the gout group compared with the other two groups. The biochemical significance of the changes in enzymic activities of the two enzymes in primary gout is discussed. PMID- 6295665 TI - Hepatic storage of bis(monoacylglycerol) phosphate without concomitant storage of sphingomyelin in a 72-year-old patient with a partial deficiency of sphingomyelinase. AB - A 72-year-old patient with marked splenomegaly and low sphingomyelinase (6% of lowest control value) in peripheral blood leukocytes is described. Much higher but variable residual sphingomyelinase activity was observed in cultured skin fibroblasts (40-67% of lowest control value); reduced activity was also found in a liver biopsy sample. Excess storage of sphingomyelin was not observed in a liver biopsy; instead, a lipid tentatively identified as bis(monoacylglycerol) phosphate was present in amounts at least 20 times greater than in age-matched control livers. The biochemical relationship of this patient to patients with sphingomyelin storage disease (Niemann-Pick disease) and phospholipidosis Type II is discussed. PMID- 6295666 TI - Transmission of incontinentia pigmenti from mother to son is consistent with a half chromatid back-mutation (reversion) model. PMID- 6295667 TI - Treatment of chronic dermatophyte infections. The use of ketoconazole in griseofulvin treatment failures. PMID- 6295668 TI - Ketoconazole therapy for pityriasis versicolor. PMID- 6295669 TI - Pox virus infection in Darier's disease. PMID- 6295670 TI - Antibodies to EB virus- and cytomegalovirus-induced antigens in early rheumatoid disease. PMID- 6295671 TI - Interaction of Epstein-Barr virus with leukaemic B cells in vitro. I. Abortive infection and rare cell line establishment from chronic lymphocytic leukaemic cells. AB - Leukaemia B cell populations, each with an individual pattern of monoclonal surface immunoglobulin expression, were obtained from 23 patients with chronic lymphocytic leukaemia (CLL) and, following exposure to a potent dose of Epstein Barr (EB) virus in vitro, were monitored for expression of the virus associated nuclear antigen EBNA, for activation of immunoglobulin synthesis and for virus induced transformation to an established cell line. Although possessing the EB virus receptor, CLL cells were generally refractory (vis-a-vis normal adult B cells) to the full effects of the viral infection. All the leukaemic populations tested developed a small proportion of EBNA positive cells within a few days post infection, but in most instances this disappeared with no subsequent evidence of viral activity. In certain cases, however, the EBNA staining became more intense, involving a larger fraction of the population and persisting for some weeks, but again this was not accompanied by virus-induced immunoglobulin synthesis or transformation. In contrast, the leukaemic cells from a single patient, tested on three separate occasions, regularly responded to EB virus infection with the rapid establishment of an EBNA positive B cell line in which the restricted pattern of surface and cytoplasmic immunoglobulin expression (gamma lambda) exactly matched that present on the original leukaemic cells. PMID- 6295672 TI - Further studies on an immunoenzymatic assay for herpes simplex virus tumor associated antigen. PMID- 6295673 TI - In vitro effects of antitumor antibody--chemotactic factor complexes. PMID- 6295674 TI - Lights and shadows on the pathogenesis of uremic polyneuropathy. PMID- 6295675 TI - Severe tubulo-interstitial disease in a renal allograft due to cytomegalovirus infection. AB - A 34 year old female developed impaired function of her renal allograft 21 months post-transplant. This was associated with lethargy, pyrexia, tenosynovitis, pancytopenia and a colonic ulcer. Severe tubulo-interstitial changes with intranuclear inclusion bodies and intracytoplasmic herpes type viral particles were seen on renal biopsy. There was no evidence of rejection. Cytomegalovirus was cultured from the urine and there was a rise in CMV antibody titer. These findings suggested the renal impairment was due to a direct cytopathic effect of the CMV. Despite treatment with transfer factor and adenine arabinoside, there was progressive loss of graft function. PMID- 6295676 TI - [Ethylene oxide polyneuropathy. Report of 2 cases with biopsy studies of nerve and muscle]. PMID- 6295677 TI - [Lipidosis with vertical gaze palsy, macular degeneration, and sphingomyelinase deficiency]. PMID- 6295678 TI - H-reflex studies in a family with possibly X-linked neuronal Charcot-Marie-Tooth disease. AB - The neuronal type of Charcot-Marie-Tooth disease (hereditary motor and sensory neuropathy, type II) was found to be present in ten of thirty-two members of one family. Motor nerve conduction velocities in this type of the disease may be normal in persons who are only mildly affected. Hoffmann-reflex investigation was effective in discriminating between affected and non-affected subjects. The mode of inheritance seemed to be X-linked, but this remained doubtful, because the outcome of the Hoffmann-reflex investigations was abnormal in one clinically non affected boy. Xg(a)-typing gave no information as to linkage with the gene-locus of CMT. Close linkage between the loci for deutan anomaly and for CMT appeared to be very unlikely. PMID- 6295679 TI - The head retraction reflex--its specificity in Parkinson's disease. AB - The head retraction reflex consists of a brief, involuntary extension of the neck on tapping the upper lip. This reflex was found to be present in 17.2% of patients with Parkinson's disease, but in only 4.9% of normal controls and 8% of cases of senile dementia. In addition there was a correlation between the severity of the Parkinson's disease and the presence of the reflex. This suggests that the reflex is specifically related to the Parkinson's disease process rather than to aging or the presence of dementia. PMID- 6295680 TI - Butterfly vertebra. AB - A patient with a so-called 'butterfly vertebra' is presented. Radiography, including computed tomography, is shown. The embryology and pathogenesis of the vertebral column with respect to vertebral abnormalities are discussed, with special reference to butterfly vertebra. Finally, attention is paid to the clinical manifestations. PMID- 6295682 TI - The Moya-Moya syndrome: a report of two children. AB - Two patients with Moya-Moya syndrome are reported. The first patient has a trisomy 21 and is to our knowledge the third patient reported with the association of Down's syndrome and Moya-Moya. The second patient also demonstrates the typical Moya-Moya picture in angiography. Angiographic follow-up is demonstrated. The presenting age and symptoms, the symptoms in the course of the disease and the prognosis of 120 non-mongoloid cases of Moya-Moya are reviewed. PMID- 6295681 TI - Ictal confusion--an electroclinical syndrome report of three adult cases with unusual features. AB - Ictal confusion--an electro-clinical syndrome consisting of varying degrees of confusion with an EEG correlate of seizure activity, abolished by intravenous diazepam, is reported in three adult patients. The unusual features were 'de novo' occurrence in adults, a chronic course, focal neurological deficits and a poor response to anticonvulsants. PMID- 6295683 TI - Central neurofibromatosis. AB - Two patients with central form of neurofibromatosis are reported. Bilateral acoustic neuromas were diagnosed by computerized tomography (CT) scan in one and at autopsy in the other patient. The clinical and radiologic features in the first patient indicated associated bilateral optic nerve gliomas. The autopsied case in addition had multiple meningiomas and numerous nontumorous glial hamartomas. The clinical, pathologic and biochemical basis for classification of neurofibromatosis into central and peripheral forms are discussed. PMID- 6295684 TI - The Hammersmith clinical experience with nuclear magnetic resonance. PMID- 6295685 TI - The role of pituitary gonadotrophin-releasing hormone receptors in the physiological regulation of gonadotrophin secretion. PMID- 6295686 TI - The influence of different types of omega 3 polyunsaturated fatty acids on blood lipids and platelet function in healthy volunteers. PMID- 6295687 TI - Hepatic clearance of serum glycoproteins. PMID- 6295688 TI - Effect of aldosterone on the human erythrocyte sodium--potassium pump in vitro. AB - 1. The effects of aldosterone in vitro on the Na+, K+-dependent ATPase activity of isolated human erythrocyte membranes and on rubidium (86Rb) uptake and [3H]ouabain binding of intact erythrocytes were studied. 2. ATPase activity was nearly doubled (0.061 +/- 0.006 to 0.110 +/- 0.01 mumol of Pi h-1 mg-1 of protein) by the addition of a physiological concentration of aldosterone (2.7 X 10(-10) mol/l). Higher concentrations had no greater effect. 3. Aldosterone had no significant effect on 86Rb uptake or [3H]ouabain binding. 4. Erythrocytes contain aldosterone at concentrations similar to that in plasma. The effect of aldosterone on ATPase is probably maximal. PMID- 6295689 TI - Recovery of infused [14C]bicarbonate as respiratory 14CO2 in man. PMID- 6295690 TI - Platelet alpha 2-adrenergic receptors in treated and untreated essential hypertension. AB - 1. One hypothesis to account for altered adrenergic response in hypertensive patients is alteration in adrenergic receptors on responsive cells. We therefore used radioligand binding methods to examine the alpha 2-adrenergic receptors on platelets isolated from 17 normal men and from 19 men with essential hypertension. In these studies we used the alpha 2-selective radioligand [3H]yohimbine to determine receptor number and affinity on intact platelets. 2. The median number of receptors per platelet was 265 for the hypertensive patients versus 246 for the platelets of controls. Likewise there was no difference between hypertensives and controls in the dissociation constant of the receptors for [3H]yohimbine or adrenaline. 3. Anti-hypertensive treatment with the alpha 2 agonist guanabenz or the beta-antagonist propranolol did not change the number or affinity of platelet alpha 2-receptors. 4. In untreated hypertensives the receptor number did not correlate with age, blood pressure, or 24 h urinary excretion of catecholamines or Na+. 5. We conclude that neither hypertension nor anti-hypertensive treatment alters the number of alpha 2-adrenergic receptors on human platelets. Furthermore, because therapy with an alpha 2-agonist does not alter the receptor number and because the receptor number did not correlate with urinary excretion of catecholamines (an index of sympathetic-nervous-system activity), we conclude that 'down-regulation' of human platelet alpha 2 adrenergic receptors may not readily occur in vivo. PMID- 6295691 TI - Effects of dietary sodium on circadian rhythm and physiological responses of 18 hydroxycorticosterone. AB - 1. The effects of dietary sodium intake on plasma 18-hydroxycorticosterone (18 OHB) responses to physiological stimuli and recumbent 24-h-plasma 18-OHB levels have been examined in nine normal male subjects. 2. Basal supine levels of 18-OHB during a 40 mmol of sodium intake period (62.5 +/- 6.0 ng/dl) were considerably greater (P less than 0.0001) than the levels during a 200 mmol of sodium intake period (9.8 +/- 1.2 ng/dl). Further incremental and percentage changes of 18-OHB in response to graded dose infusions of angiotensin II and adrenocorticotropic hormone (ACTH) were greater during the 40 mmol of sodium intake period. 3. Although the mean 24 h levels of plasma 18-OHB during the 40 mmol of sodium intake period (43.9 +/- 4.0 ng/dl) were greater (P less than 0.001) than those during the 200 mmol of sodium intake period (9.4 +/- 1.2 ng/dl), the circadian rhythm of 18-OHB secretion was similar under the two extremes of sodium intake. 4. Factors which increase angiotensin II levels, such as sodium restriction, isometric exercise and angiotensin infusion, selectively increase 18-OHB and aldosterone, suggesting that angiotensin II increases 18-OHB and aldosterone secretion, in part, by modulation of the 18-hydroxylation reaction involved in conversion of corticosterone into 18-OHB. PMID- 6295692 TI - Elevated arterial cyclic AMP levels during the onset of renal hypertension in rats. AB - 1. To determine the possible role of arterial cyclic AMP in the pathogenesis of hypertensive vascular hypertrophy and hyperplasia, the changes in the level of this nucleotide were studied during the development of renal hypertension in rats with aortic ligation between the renal arteries. 2. A twofold increase in the cyclic AMP level of the thoracic aorta was observed in 9-day hypertensive rats when compared with sham-operated controls. At this time the total amounts of DNA and collagen were unchanged, although a marked increase in arterial fibrous protein was already present. 3. Arterial cyclic AMP remained significantly elevated in the thoracic aorta of 30-day hypertensive animals. At this time the hypertensive vascular alterations had reached completion as shown by the abnormal accumulation of collagen, DNA and non-fibrous protein. 4. Contrary to the events taking place in the thoracic aorta, a marked decrease in cyclic AMP was present in the abdominal portion, which was protected from high blood pressure by the aortic ligature. In this segment decreased cyclic AMP coexisted with an unchanged collagen content and a diminution in the contents of DNA and non-fibrous protein. 5. Thus a marked increase in arterial cyclic AMP precedes the initiation of DNA replication and collagen accumulation in vascular territories subjected to high blood pressure. These studies suggest the participation of this nucleotide in the vascular growth induced by hypertension. PMID- 6295693 TI - [Effects of suloctidil therapy on platelet function in patients with on obliterating arteriopathy of the lower limbs]. PMID- 6295694 TI - Modulation of type I and type III collagen production in normal and mutant human skin fibroblasts by cell density, prostaglandin E2 and epidermal growth factor. AB - Previously, we found that, at high cell density, human skin fibroblasts produce a higher proportion of type III to type I collagen. Treatment of sparse cultures with prostaglandin E2 also increases this ratio, while treatment of dense cultures with epidermal growth factor reduces it. Similar changes were observed using some strains of fibroblasts from patients with the Ehlers-Danlos syndrome type IV and osteogenesis imperfecta. Studies on the extent of intracellular degradation suggest that the changes observed could be caused in part by changes in the breakdown of collagen inside the cell with preferential loss of type I collagen and/or changes in synthesis of the two proteins. PMID- 6295695 TI - Production and characterization of hybridoma antibody to native human type II collagen. AB - Using a sensitive RIA for collagenous proteins, we have applied a series of consecutive selection steps to construct a hybridoma that produces antibody against native human Type II collagen. The selection steps consisted of: 1) hybrid cell line selection; 2) co-selection of IgG class immunoglobulin reactive to the immunizing antigen; 3) selection for anticollagen antibody by complete inhibition of antibody binding to Type II collagen-coated plates by prior specific collagenase digestion; 4) selection for anti-helical antibody by complete inhibition of antibody binding to heat-denatured Type II collagen-coated plates; 5) clonal selection; 6) selection for absence of crossreactivity to 5-7 additional native collagen types; and finally 7) selection for antibody yielding indirect immunofluorescent staining of human cartilage matrix, but not adjacent bone or perichondrium. Cross-reactivity assays against other native human collagen types revealed little cross-reactivity. In addition, we have demonstrated that native Type II collagen inhibits antibody binding to Type II collagen-coated plates; that Type II collagen can be recovered from antigen antibody complexes; that a sensitive radioimmunoassay for Type II collagen can be developed; and that the antibody is useful for indirect immunofluorescent studies of human cartilage matrix. PMID- 6295696 TI - Development cues in limb bud chondrogenesis. PMID- 6295697 TI - Isolation and characterization of the mitochondrial DNA from the florida spiny lobster, Panulirus argus. AB - 1. Mitochondria and mitochondrial DNA were isolated from the digestive gland of Panulirus argus. 2. The mitochondrial DNA has an average contour length of 5.13 microns which corresponds to a molecular weight of 10.10 X 10(6) daltons. 3. The molecular weight based on agarose gel electrophoresis of restricted individual DNA samples ranges from 10.04 to 10.4 X 10(6) daltons. 4. Restriction endonuclease analysis with Bam H1 and Eco R1 demonstrate variation in nucleotide sequence between individual lobsters. PMID- 6295699 TI - Vitamin B12 binders (transcobalamins) in serum. AB - The measurement of plasma or serum vitamin B12 (B12) binders is important in the diagnosis of congenital megaloblastic anemias, the myeloproliferative disorders and perhaps in other malignancies. Two classes of binders circulate in plasma, Transcobalamin II (TC II) and R-binders. The latter have been divided by some authors into transcobalamin I (TC I) and transcobalamin III (TC III), and the validity of this division is discussed. R-binders and TC II differ in their apparent molecular weight on gel filtration chromatography, by which method they can be separated reliably. However, the technique is time-consuming and cumbersome and a variety of rapid separation methods have been described in the literature. These are discussed and compared to the standard gel filtration separation method. TC I and III are separable on the basis of their differing charges, and the methods which have been described for accomplishing this are compared and critically reviewed. There has been some controversy in the literature as to whether plasma or serum should be used to measure circulating B12 binders. Leukocytes, which contain R-binders, release these in vitro and this release is greatest when blood is allowed to clot and the serum separated. Consequently, the use of plasma, sometimes with agents added to prevent leukocytic release of binder, has been advocated. Yet, none of the agents used eliminates artifact totally, and this aspect, too, is reviewed. Lastly, several techniques for the purification of B12 binders have been described. Some techniques result in pure binder preparations, while others result in preparations which are free of other binders but contaminated with non-B12 binding proteins. Both approaches have advantages and disadvantages, and these are discussed. PMID- 6295698 TI - The conformation and molecular biology of pancreatic hormones and homologous growth factors. PMID- 6295700 TI - Immunologic aspects of melanoma. PMID- 6295702 TI - Intraocular pressure following systemic administration of cannabinoids. AB - Various cannabinoids have been tested for activity compared to delta 9-THC in reducing intraocular pressure after intravenous administration in rabbits at 0.1 mg or 1 mg/animal. Comparison of l-delta 9-, delta 8-, 11-OH-delta 9- and 11-OH- delta 8-THC indicates that minor configurational changes have only a small influence on activity with regard to induction of a fall in intraocular pressure, although 11-OH-delta 8-THC has increased activity. 8 alpha-OH-, 8 alpha-diOH- and 8 beta-diOH-delta 9-THC have little or no activity but 8 beta-OH-delta 9-THC is as active as delta 9-THC indicating that the hydroxyl group in the beta-position does not influence activity. Modification of the C5H11 alkyl side chain (3'-OH delta 9-THC) reduced activity to 20% relative to delta 9-THC. Cannabidiol (CBD), cannabichromene, cannabigerol and olivetol had no activity, but 10-OH-CBD had some activity at 2 mg/animal. Cannabinol (CBN) had about half the activity of delta 9-THC and activity was reduced further with 1'-OH-CBN, indicating that side chain modification reduced activity. Neither delta 9-THC, nor cannabigerol, had any effect on intraocular pressure or total outflow facility in the rhesus monkey, suggesting species differences in ocular responses to cannabinoids. Further studies on modification of these compounds is warranted in order to further delineate the structure-activity relationships. PMID- 6295701 TI - Cytotoxic effects of calcium on sodium-potassium transport in the mammalian lens. AB - Experiments were designed to investigate the effects of lens calcium on the Na/K pump since elevated calcium and sodium often accompanies lens opacification. An increase in lens calcium was accomplished by culturing lenses in calcium-rich media at 21 degrees C in the absence of poisons which might directly alter sodium transport. Calcium loaded lenses were subsequently incubated at 37 degrees C in a normal culture medium to assess membrane transport function. A five-fold increase in total lens calcium to 1.45mM, resulting in a 4% increase in membrane bound calcium, had little effect on the cation pump. However, an increase in lens calcium by a factor of 100 caused a 4-fold increase in membrane bound calcium, a 50% decrease in (Na/K)-ATPase activity, and a 40% decrease in 86Rb uptake after an incubation period of 20 hrs. Details of the mechanism of enzyme inhibition remain obscure, but the data suggest the inhibition is predominantly irreversible and does not involve an interaction between calcium and ATP. PMID- 6295704 TI - Bronchiolo-alveolar cell carcinoma diagnosed by bronchoalveolar lavage. PMID- 6295705 TI - Central alpha-adrenoceptors: clinical applications in cardiovascular disease. PMID- 6295703 TI - Role of glutathione in the prevention of cataractogenesis in rat lenses. AB - A new method has been developed for the conjugation of rat lens glutathione by 1 chloro-2,4-dinitrobenzene (CDNB). This reaction is catalysed by glutathione S transferase present in the lens. One milliliter of 1 mM CDNB per two rat lenses conjugates more than 95% of the lens GSH in 30 min at 37 degrees. Lenses incubated with 1 mM CDNB for 30 min, followed by incubation in the culture medium without CDNB remained apparently clear for up to 24 hrs. The CDNB treated lenses were more susceptible to protein precipitation (cataract formation) when challenged with oxidants such as hydrogen peroxide and superoxide anions. PMID- 6295706 TI - Alpha-adrenoceptors, adrenaline, and exaggerated vasoconstrictor response to stress in essential hypertension. AB - Stressful sympathetic stimulation by cold pressor test in patients with essential hypertension results in an exaggerated response of the already elevated plasma adrenaline, heart rate, blood pressure, and alpha-adrenoceptor-mediated vasoconstriction when compared with normotensive subjects. The stress-induced increase in adrenaline was correlated with the attendant increase in blood pressure. The stress-induced reduction in forearm flow was reversed during infusion of the postjunctional alpha 1-adrenoceptor blocker prazosin. Therefore, enhanced responses to sympathetic stress, as reflected and perhaps caused by an exaggerated rise in plasma adrenaline, may contribute to an increased alpha 1 adrenoceptor-mediated vasoconstriction in essential hypertension. PMID- 6295707 TI - Role of opioid peptides in brain mechanisms regulating blood pressure. AB - Beta-endorphin and related opioid peptides are neuropeptides which appear to play a role in cardiovascular regulation which is supported by altered nociceptive responsiveness in hypertensive animals. In spontaneously hypertensive rats the pain threshold for electric stimulation is elevated; these rats show increased response latency time in a hot plate test. The opiate antagonist naloxone reverses these values to that of the normotensive controls. In other forms of experimental hypertension, eg, renal hypertension (one-clip, two-kidney model), no change in pain sensitivity is apparent. Sinoaortic baroreceptor denervation causes a labile hypertension without changes in hot plate response. Administration of beta-endorphin into the nucleus of the solitary tract (NTS) gradually decreases blood pressure and heart rate without affecting respiratory frequency. These cardiovascular effects are blocked by naloxone as well as by an antibody to beta-endorphin. In contrast to the effects of beta-endorphin, microinjection of enkephalins into the NTS increases blood pressure and heart rate. The data suggest the existence of two separate endorphin systems at the level of the NTS, one a depressor and another a pressor system. The depressor influence of beta-endorphin may play a role in the mechanism of action of antihypertensive agents such as methyldopa and clonidine. Our data support a role of endorphins as neuropeptides involved in cardiovascular regulation, exerting a dual influence at the level of the NTS. PMID- 6295708 TI - Central alpha-adrenergic control of blood pressure. Effects of clonidine withdrawal. AB - Studies were undertaken to investigate the role of the CNS in withdrawal hypertension caused by abrupt cessation of long-term clonidine treatment. The studies were conducted on male wistar rats receiving clonidine, 100 micrograms/kg subcutaneously twice daily for seven days. Withdrawal hypertension occurred 16 to 18 hours following the last injection of clonidine. Pressor responses to posterior hypothalamic stimulation were significantly potentiated in the rats undergoing clonidine withdrawal, and the depressor response to a single injection of 10 micrograms of clonidine into the lateral cerebral ventricle was significantly attenuated in these animals. Studies on pithed rats demonstrated that destruction of the CNS eliminated the withdrawal hypertension. These data suggest that alpha-adrenoreceptors in the nucleus tractus solitarius play an important role in the normal maintenance of blood pressure, and a decrease in sensitivity of these receptors could result in the development of a hypertensive state. PMID- 6295709 TI - Pharmacology and characterization of central alpha-adrenoceptors involved in the effect of centrally acting antihypertensive drugs. PMID- 6295710 TI - Characterization of the clonidine receptor site. AB - At the cardiac sympathetic nerve terminal the alpha 2-adrenoceptor is presynaptic and appears to be located at an extrasynaptic site. This is suggested by (1) absence of evidence of autoinhibitory feedback at physiologic stimulus levels up to about 50 percent of the maximum chronotropic response in the isolated guinea pig right atrium, and (2) absence of significant competition between clonidine and synaptically released noradrenaline (NA) for the presynaptic site. In the central nervous system (CNS) cardiovascular alpha 2-receptors are probably located at a postsynaptic site in bulbospinal regions of the brain, since they produce effects identical to those of synaptic release of NA. Experiments with the clonidine analog alinidine (ST 567) suggest that there are differences in central receptor type subserving clonidine-mediated baroreflex heart rate and blood pressure changes. PMID- 6295711 TI - Relationship of immunoassayable clonidine plasma levels to its pharmacologic action in clinical and experimental hypertension. PMID- 6295712 TI - Comparative quantitative studies on central and peripheral alpha-adrenoceptors. PMID- 6295713 TI - Interactions of leucine-enkephalin with alpha-adrenoceptors in the conscious dog. AB - Leucine-enkephalin (Leu-ENK) administered systemically to the conscious dog increases heart rate and blood pressure, a response prevented by naloxone and hence mediated by opiate receptors. Clonidine attenuates the heart rate increase without affecting the pressor response. Propranolol also reduces the magnitude of the heart rate increase but to a lesser degree than does clonidine. Clonidine appears to inhibit the Leu-ENK-induced heart rate increase by both a centrally mediated increase in vagal tone and a peripheral presynaptic alpha 2-agonist effect. The presence of a pressor response to Leu-ENK after prazosin pretreatment suggests that Leu-ENK may also increase blood pressure independently of the alpha 1-adrenoceptor. Thus, Leu-ENK appears to act via a naloxone-sensitive receptor (perhaps a vagal afferent chemoreceptor), the response involving both sympathetic and parasympathetic mechanisms. The pressor response may also involve a direct effect on peripheral vessels or a non-alpha 1-adrenergic efferent pathway. PMID- 6295714 TI - Effects of long-term clonidine administration on the hemodynamic and neuroendocrine postural responses of patients with dysautonomia. AB - Patients with mitral valve prolapse syndrome (MVPS), vasoregulatory asthenia, and poor postural adjustment often have orthostatic intolerance characterized by tachycardia and a narrow pulse pressure on standing. Autonomic dysfunction is thought to play an important role. Increased alpha-adrenergic activity has been shown in MVPS patients with orthostatic intolerance. We measured hemodynamic and neuroendocrine responses to long-term oral clonidine therapy in eight women, aged 36 +/- 1.8 years (27 to 44 years). None had responded favorably to beta-blockers. Heart rate, blood pressure, oxygen consumption, cardiac output, and plasma norepinephrine levels were measured in both supine and standing positions, before and after one to four weeks of clonidine (0.3 to 0.4 mg daily). Clonidine reduced standing plasma norepinephrine levels, total peripheral resistance, and diastolic blood pressure; a smaller decrease in cardiac output on standing was noted. Plasma volumes increased 12 percent. Mild reductions in plasma catecholamines and total peripheral resistance are associated with fewer, not more, orthostatic symptoms in this group of patients. "Placebo" or mild sedative effects may explain part of the response to clonidine, but the hemodynamic and neuroendocrine data suggest that decreased alpha-adrenergic hyperactivity may also be important. PMID- 6295715 TI - Selective angiography in diagnosing primary hepatic carcinoma. PMID- 6295716 TI - [Malignant mixed mesodermal tumors of the uterus: a clinicopathologic report of 12 cases]. PMID- 6295717 TI - [Alteration of cAMP and cGMP levels in the peripheral blood leukocytes of acute leukemic patients]. PMID- 6295718 TI - The Golgi apparatus: roles for distinct 'cis' and 'trans' compartments. AB - The Golgi apparatus seems to consist of distinct cis and trans compartments that are proposed to act sequentially to refine the protein export of the endoplasmic reticulum by removing escaped endoplasmic reticulum proteins. Refinement may be a multi-stage process that employs a principle akin to fractional distillation; the stack of cisternae comprising the cis Golgi may be the plates in this distillation tower. The trans Golgi, consisting of the last one or two cisternae, may be the receiver that collects from the cis Golgi only its most refined fraction for later distribution to specific locations throughout the cell. PMID- 6295719 TI - Steps in the phosphorylation of the high mannose oligosaccharides of lysosomal enzymes. AB - The phosphomannosyl recognition marker of acid hydrolases, which mediates their translocation to lysosomes, has been shown to be synthesized in two steps. First, N-acetylglucosamine 1-phosphate is transferred to an acceptor mannose by UDP-N acetylglucosamine:lysosomal enzyme N-acetylglucosamine-1-phosphotransferase, resulting in a phosphate group in diester linkage between the outer N acetylglucosamine and the inner mannose. Next, an a-N-acetylglucosaminyl phosphodiesterase removes the N-acetylglucosamine, leaving the phosphate in monoester linkage with the underlying mannose residue. This exposed phosphomannosyl residue serves as the essential component of a recognition marker which leads to binding to high-affinity receptors and subsequent translocation to lysosomes. We propose that the first enzyme in this scheme, N acetylglucosaminylphosphotransferase, catalyses the initial, determining step by which newly synthesized acid hydrolases are distinguished from other newly synthesized glycoproteins and thus are eventually targeted to lysosomes. The absence of this enzyme activity, as in inclusion-cell (I-cell) disease and pseudo Hurler polydystrophy, precludes the receptor-mediated targeting of newly synthesized acid hydrolases to lysosomes. As a consequence, the enzymes are secreted into the extracellular milieu. PMID- 6295721 TI - Endocytosis, the sorting problem and cell locomotion in fibroblasts. AB - Fibroblasts endocytose lipid plus a subset of plasma membrane proteins over their entire surface and reinsert this into the plasma membrane at the cell's leading edge. This process is used to extend the fibroblast forwards. This circulation causes a flow of these endocytosed molecules over the cell's surface. Molecules, such as proteins, sitting in this flow can distribute themselves randomly by Brownian motion, but large objects (or small tethered ones) cannot. These large objects therefore cap. A mechanism is presented whereby this process could be used for locomotion using many weak interactions with the substrate. In addition it is suggested that the observed selectivity of coated pits may be sufficient to sort out proteins during transfer of membrane from one organelle to another so that the specific characters of the parent membranes are maintained. PMID- 6295720 TI - Receptor-mediated transport of IgG across the intestinal epithelium of the neonatal rat. AB - The absorptive epithelium of the neonatal rat is developmentally specialized to transfer maternal immunoglobulin G (IgG) intact to the circulation while other milk protein are digested. The epithelial cells of the duodenum and proximal jejunum which are responsible for IgG transfer represent a particular striking experimental model for study of receptor-mediated intracellular transport. Receptors located on the luminal plasma membrane selectively bind the Fc region of IgG. The IgG enters the cell by constitutive endocytosis within coated vesicles and is then released at the basolateral plasma membrane. Morphological evidence supports a model in which IgG crosses the cell as a ligand-receptor complex that dissociates only on exposure to a pH 7.4 environment found at the basolateral cell surface. Although uptake of IgG at the luminal plasma membrane is highly selective, small but significant amounts of other proteins enter the cell apparently non-selectively. Nevertheless, these latter proteins are not transferred across the cell. Double-tracer experiments indicate that IgG and these other proteins enter the cell simultaneously within the same endocytic vesicles, but that non-membrane-bound proteins are removed from the IgG transport pathway by an as yet poorly defined mechanism and sequestered within small apical vacuoles and lysosomes. PMID- 6295722 TI - Epidermal growth factor: uptake and fate. AB - Lateral diffusion of epidermal growth factor (EGF) receptors along the plane of the cell membrane can be measured using fluorescently labelled analogues of EGF and the fluorescence photobleaching recovery method in cultured cells. With the aid of high image-intensified fluorescent microscopy, the receptors, which are initially distributed diffusely, form patches and undergo endocytosis at 37 degrees C. These gross processes may not be critical in mediating the initial, rapid actions of the hormone. The processes of uptake and endocytosis correspond biochemically to the loss of surface receptors ('down-regulation') and degradation of the receptor and hormone via lysosomes. The EGF receptors are not apparently recycled or re-utilized, and they are continuously internalized, even in the absence of ligand. Since all manoeuvres that interfere with intracellular degradation or processing block mitogenesis, it is proposed that one or both of these may be essential processes, although in such a case they must be continuous and protracted functions. Slow nuclear accumulation of the complex of hormone and receptor may be an important process. In addition, evidence suggests that limited (submicroscopic) receptor aggregation (dimerization) at the cell surface may be necessary and sufficient to trigger the long-term effects (but not the immediate effects), and thus this aggregation may be required for endocytosis. The ligand itself may not be an essential structural component of the action of the receptor since anti-receptor antibodies can elicit mitogenic responses. Recent results suggest that EGF receptors normally exist in a low affinity state which is rapidly converted by EGF (at 37 degrees C but not at 4 degrees C) to a high affinity state by a process that requires prior intact protein synthesis. Furthermore, the accumulation of a special, stable intracellular pool of the complex may be related to the control of cellular growth (and tumour promotion). PMID- 6295723 TI - [Hepatoblastoma in children]. PMID- 6295724 TI - Profiles of intermediary metabolites in insulin-dependent pregnant diabetic women with or without endogenous insulin production. AB - The levels of glucose, free fatty acids (FFA), glycerol, 3-hydroxybutyrate (3 HB), cyclic AMP, and C-peptide immunoreactivity (CPR) were determined in venous plasma samples taken every half hour during an 8-h period under standardized conditions in 10 insulin-dependent pregnant women. The metabolic profiles were determined in each trimester of pregnancy. The women were divided into two groups, one with (group I: 3B, 1C, and 1D according to White's classification) and one without (group II: 1B, 3C, and 1D) measurable plasma CPR levels. Plasma glucose values were significantly higher (P less than 0.01) with greater variability in group II than in group I in the first and second trimesters, while no such difference was found in the third trimester of pregnancy. FFA, glycerol, 3-HB, and cyclic AMP levels were not different between the groups in any trimester of pregnancy. Amniotic fluid CPR was higher and skinfold thickness of the newborn greater in group II than in group I. Neonatal complications occurred only in infants of group II mothers. It is concluded that determination of plasma CPR in diabetic women in early pregnancy could give additional prognostic information to that obtained by the White classification. PMID- 6295725 TI - [Properties of monoclonal antibodies to the Venezuelan equine encephalomyelitis virus]. PMID- 6295726 TI - [Correlation of the changes in the levels of the low-spin oxidized form of cytochrome P-450 and in the intensity of protein biosynthesis]. PMID- 6295727 TI - [Secondary changes of recombinant DNA in the cloning of the early region of the phage T7 genome]. PMID- 6295728 TI - [Initiation of RNA synthesis by the encephalomyocarditis virus in a cell-free system and the possible participation of VPg protein in this process]. PMID- 6295729 TI - [Replication of hybrid plasmids containing polyoma virus genes in a yeast cell]. PMID- 6295730 TI - [ATP receptor in the membrane of sensory neurons]. PMID- 6295731 TI - [Determination of nucleosome configuration along the DNA by a molecular cloning method: mapping of the core-particle positions in the minichromosome of simian virus 40]. PMID- 6295732 TI - [Isolation of a plasmid clone containing the mRNA sequence for mouse nonviral T antigen]. PMID- 6295733 TI - [Transposon "memory" of a previous integration site]. PMID- 6295735 TI - [Benzodiazepine receptors of the mouse glioblastoma cell line: the anomalous effect of gamma-aminobutyric acid on diazepam binding]. PMID- 6295736 TI - Cyproheptadine-induced remission of Cushing's disease due to pituitary basophil adenoma. AB - Serotonin is involved in the control of ACTH secretion, possibly by stimulating corticotropin releasing factor secretion from the hypothalamus. Cushing's disease seems to be due to defective hypothalamic regulation of ACTH release from the pituitary gland. Cyproheptadine is a potent antagonist of serotonin and has been used successfully in some patients with Cushing's disease, although, generally, in women without radiological evidence of pituitary tumors. We report the successful use of cyproheptadine in a 54-year-old man with Cushing's disease due to pituitary basophil adenoma. Significant clinical and biochemical improvement was noted 45 days after treatment began. The results in this patient support our findings that cyproheptadine can be effective in patients with Cushing's disease due to pituitary tumors, as well as in preparing very ill patients for surgery or managing such patients until radiotherapy takes effect. PMID- 6295734 TI - [Transformation of Chinese hamster cells by plasmid pBR325::tk::HS3 and the effect on transformant stability of tumor promotor TPA]. PMID- 6295737 TI - Low density lipoprotein receptors in adrenal tissue of a human anencephalic fetus. AB - The binding of [125I]iodoLDL and [125I]iodoHDL to membrane fractions prepared from fresh adrenal tissue of an anencephalic newborn was examined and compared to the binding of these lipoproteins to membrane fractions prepared from normal human fetal adrenal (HFA) tissue. The apparent dissociation constant (KD) for the interaction of [125I]iodoLDL with its binding sites in membrane fractions prepared from HFA tissue of an anencephalic fetus (23.5 micrograms . ml-1) was similar to that in membranes from normal HFA tissue. However, the binding capacity for [125I]iodoLDL of membrane fractions prepared from adrenal tissue of an anencephalic fetus (600 ng . mg-1 protein) was only one-third of the binding capacity (1500 ng . mg-1 protein) of normal HFA membrane fractions. On the other hand, the binding capacity for [125I]iodoHDL of membrane fractions prepared from anencephalic and normal adrenal tissue was similar. PMID- 6295738 TI - [Differences in the damaging action of tetrachloromethane on the protein synthesizing capacity and the glucose-6-phosphatase activity of rat liver microsomes]. PMID- 6295739 TI - [Electrophysiologic changes in thalidomide neuropathy in the treatment of discoid lupus erythematosus]. AB - Twenty-six patients suffering from lupus erythematodes discoides were under clinical and electrophysiological examination. The results are in agreement with the concept of a mainly sensory, distal and axonal polyneuropathy. A correlation between total dosage and severeness of polyneuropathy could not be found. PMID- 6295740 TI - Electromyography, single fibre electromyography and necropsy findings in myasthenic syndrome associated with bronchogenic carcinoma. PMID- 6295741 TI - Influence of the interstimulus interval on the extraterritorial potentials of the motor units. PMID- 6295742 TI - The recovery of hormone responsiveness in desensitized neonatal rat calvariae. AB - Two issues were studied: the loss of hormone responsiveness during culture of neonatal rat calvariae under standard culture conditions and the recovery of hormone responsiveness in calvariae previously desensitized with PTH (2.5 U/ml), prostaglandin E2 (PGE2; 1 or 2.5 micrograms/ml), or salmon calcitonin (sCT; 100 mU/ml). Both of these issues were studied in the presence and absence of cortisol (10(-6) M). Culture of neonatal rat calvariae in medium without cortisol resulted in a significant decrease in cAMP responsiveness to PTH, PGE2, and sCT. The addition of cortisol to the culture medium: 1) maintained cAMP responsiveness to PTH at levels approximately 50% of the response of freshly dissected, uncultured calvariae, 2) increased cAMP responsiveness to PGE2 to 120% of the fresh response, and 3) enhanced the loss of cAMP responsiveness to sCT. With regard to the recovery of hormone responsiveness in calvariae previously desensitized with PTH, PGE2, or sCT and then cultured in medium with or without cortisol, the following observations were made. 1) PTH-desensitized calvariae required a 24-h culture period in cortisol to fully recover cAMP responsiveness to PTH. Without cortisol, the recovery was only partial. 2) PGE2-desensitized calvariae were fully recovered after a 24-h culture period in the presence or absence of cortisol. Further experiments indicated that recovery of PGE2-desensitized calvariae was complete after only 2 h of culture in cortisol. 3) sCT-desensitized calvariae did not regain responsiveness to sCT in the presence of cortisol, but partially recovered responsiveness in the absence of cortisol. These results indicate that the recovery of hormone responsiveness of bone in vitro after homologous desensitization with sCT, PTH, or PGE2 follows a distinct pattern for each hormone and suggest that different cellular mechanisms may be involved. PMID- 6295743 TI - Pituitary gonadotropin-releasing hormone receptors during gonadotropin surges in ovariectomized-estradiol-treated rats. AB - In intact cycling rats, the number of pituitary GnRH receptors varies markedly during the estrous cycle. Concentrations are maximal on diestrus and early proestrus, before falling rapidly for a brief period immediately before the preovulatory gonadotropin surge. In this study we investigated whether dynamic changes in ovarian steroids, pituitary hormones, and GnRH itself, all of which are changing at the time of the surge, play a role in the acute transient down regulation of the pituitary GnRH receptors. We used the ovariectomized-estradiol treated female rat as a model, as these animals exhibit daily gonadotropin surges at a predictable time of the day and also allow studies in a situation where concentrations of ovarian steroids are stable. The pituitary GnRH binding capacity (GnRH-BC) was measured using the analog D-Ala6des Gly10-GnRH ethylamide as ligand. GnRH-BC was stable between 0900-1530 h [range, 288 +/- 29 to 262 +/- 33 fmol protein (mean +/- SE)] and fell abruptly to 123 +/- 17 fmol/mg at 1630 h, before returning to the initial level by 1730 h. This abrupt fall in GnRH-BC preceded the afternoon gonadotropin surge and was similar in timing, magnitude, and duration to that observed in intact cycling rats. Serum PRL decreased from peak levels at 1630 h, coincident with the fall in GnRH-BC, before rebounding at 1730 h. Pentobarbital given at 1400 h abolished both the gonadotropin surge and the acute fall in GnRH-BC, but did not change serum PRL levels, suggesting that PRL is not causally related to the fall in GnRH-BC. The stable morning levels of GnRH-BC were not reduced after iv injections of LH, FSH, or both hormones despite elevations in serum gonadotropins to concentrations greater than those seen during the afternoon surge. Additionally, multiple iv injections of GnRH at 30- or 10-min intervals did not decrease the stable morning levels of GnRH-BC, although serum LH and FSH were markedly elevated. The data suggest that dynamic fluctuations in ovarian steroids, gonadotropins, PRL, and GnRH are not causally related to the acute transient reduction of pituitary GnRH receptors before the afternoon gonadotropin surge. These results also suggest that another hypothalamic or pituitary factor(s) is involved in the acute regulation of GnRH receptors, and the ovariectomized-estradiol-treated rat appears to be a good model for the elucidation of the factor(s) involved. PMID- 6295744 TI - Parathyroid hormone induces a calmodulin-dependent alteration in phosphodiesterase activity of rat kidney in vivo. AB - In the basal state cyclic nucleotide phosphodiesterase in cytosol of rat kidney cortex is characterized by at least two kinetically distinct activities, with Michaelis-Menten constant (Km) for cAMP of 3.3 X 10(-6) and 3.7 X 10(-5) M. Eight minutes after in vivo injection of parathyroid extract, and persisting over 4 h, phosphodiesterase showed only one apparent activity, Km = approximately 1 X 10( 5) M. This change in kinetic profile was mimicked by the ip injection of 10 mg/kg (Bu)2-cAMP, and was reversed by incubation of cytosol with trifluoperazine or EGTA. Incubation of control cytosol with purified calmodulin duplicated the effect of hormone injection. Chromatography of control cytosol over diethylaminoethyl-cellulose resolved two activity peaks. Peak I, corresponding to the high Km activity of whole cytosol, was stimulated by calmodulin, representing a change in maximum velocity but not in substrate affinity. In addition, calmodulin stimulated the hydrolysis by Peak I of 3',5'-cyclic guanosine monophosphate. Peak II activity was not altered by calmodulin. These studies demonstrate that PTH promotes a rapid and sustained alteration in phosphodiesterase kinetics which is mediated by calcium and calmodulin, and which reflects the activation of the high Km component of the enzyme. PMID- 6295745 TI - Specific binding of growth hormone to isolated chondrocytes from rabbit ear and epiphyseal plate. AB - Chondrocytes were isolated from rabbit ear, epiphyseal and rib cartilage, and their ability to bind 125I-labeled human growth hormone (hGH) was investigated. The total binding of hormone to ear chondrocytes increased with time until 4-6 h, whereas non-specific binding did not increase. Total binding was 1-3% of total counts added, and non-specific binding was 10-20% of total binding. Unlabeled hGH, bGH and oPRL competed with the binding of labeled hGH in a dose-dependent manner with half-maximal binding at concentrations of 10-20 ng/ml (hGH) and 100 ng/ml (bGH and oPRL). The two latter hormones displaced almost all specific binding of hGH at higher concentrations. Ear chondrocytes from newborn to 4 week old rabbits all showed specific binding of hGH with no clear age dependence. Rib chondrocytes, on the other hand, showed very little specific binding. Epiphyseal chondrocytes from newborn animals yielded intermediate binding values. The finding of specific hGH binding by cultured chondrocytes supports the concept of a direct effect of GH on chondrocytes. PMID- 6295746 TI - Partial isolation and characterization of testicular GnRH-like factors. AB - We report here partial isolation and characterization of at least two GnRH receptor binding factors from the ethanol: chloroform: acetic acid (ECA) extracts of rat testis. The displacement curve of defatted, steroid-free and desalted ECA extract was parallel to that of D-(leu)6-des (Gly)10-GnRH-EA in a GnRH radioreceptor assay. Immunoaffinity chromatography on cyanogen bromide-activated Sepharose 4B beads covalently bound to an antibody raised against d-(lys)6-GnRH resulted in more than a hundredfold increase in receptor binding specific activity. Equivalent amounts of kidney extract after affinity chromatography showed no significant activity. Coincubation of the material purified by affinity chromatography with the labeled ligand did not result in significant peptidase degradation of the label, indicating that apparent displacement of the label in the receptor assay was not the result of cleavage of the ligand. HPLC of the material partially purified by affinity chromatography on a reverse phase 5 micron ODS column revealed two peaks of receptor binding activity. Preliminary estimates of molecular weights of these factors based on SDS-PAGE and gel filtration are 68,000 and 6,000 respectively. We conclude that there are at least two factors in rat testis with GnRH-receptor-binding properties that are chemically distinct from the native decapeptide. PMID- 6295747 TI - Internalization and degradation of human chorionic gonadotropin in ovine luteal cells: effects of inhibition of protein synthesis. AB - Studies were undertaken to investigate the possibility that receptors for LH in monolayer cultures of enzymatically dissociated ovine luteal cells are recycled. Cultured cells (3-10 X 10(5) total steroidogenic cells/dish) were incubated with or without cycloheximide (CHX; 10(-4) M) and 2 X 10(6) cpm [125I]iodo-hCG in the presence or absence of 30 micrograms nonradioactively labeled hCG for 0, 12, 24, 36, or 48 h. At each time point, the amounts of radioactivity bound to the cells (bound [125I]iodo-hCG), located intracellularly (intracellular [125I]iodo-hCG), and degraded and returned to the medium as [125I]monoiodotyrosine (degraded [125I]iodo-hCG) were determined. The number of receptors for LH was determined by Scatchard analysis. Cell viability was also monitored by: 1) trypan blue dye exclusion, 2) the ability of the cells to synthesize protein, and 3) basal and hCG-stimulated secretion of progesterone. More than 90% of the cells remained viable after 48 h of culture, and CHX had no effect on cell viability. Protein synthesis in CHX-treated cells was inhibited by more than 90%. Basal and hCG stimulated secretion of progesterone were also inhibited by CHX. Treatment with CHX increased the amounts of membrane-bound and internalized [125I]iodo-hCG and decreased the amounts of [125I]iodo-hCG that were degraded. When the quantities of radioactivity in these three fractions (plasma membrane-bound, internalized, and degraded) were added together to obtain a value for the total amount of [125I]iodo-hCG that had been bound to receptor during the 48-h time course (total receptor-associated [125I]iodo-hCG), the value for control cells was not significantly different from the value for CHX-treated cells. Furthermore, the total receptor-associated [125I]iodo-hCG was approximately 2-fold greater than the amount of [125I]iodo-hCG required to saturate receptors at time zero. These data indicate that synthesis of new receptors is not required for the continued binding, internalization, and degradation of [125I]iodo-hCG. Further, the data are compatible with the hypothesis that receptors for LH are recycled or that a portion of the total receptor population is in an unavailable form when the cells are intact but are available for binding after homogenization of the cells. PMID- 6295748 TI - Inhibitory actions of a gonadotropin-releasing hormone agonist on ovarian follicle-stimulating hormone receptors and adenylate cyclase in vivo. AB - The regulation of ovarian gonadotropin-sensitive adenylate cyclase and FSH receptors was studied in hypophysectomized diethylstilbestrol-primed rats treated with FSH and/or the potent GnRH agonist [D-Ala6]des-Gly10-GnRH N-ethylamide (GnRHa). The animals were treated with 7.5 micrograms ovine FSH twice daily for 2 days, either alone or with 10 micrograms GnRHa. FSH-stimulated adenylate cyclase activity was augmented by 2.5- to 3.5-fold in the presence of 5'-guanyl imidodiphosphate. Adenylate cyclase responses to FSH were almost completely abolished by GnRHa treatment in ovarian homogenates from control animals and rats treated with FSH. This inhibition was receptor specific, since GnRHa did not block adenylate cyclase stimulation by prostaglandin E2 or isoproterenol. No inhibition of 5'-guanyl-imidodiphosphate- or sodium fluoride-stimulated adenylate cyclase activity was noted after any hormone treatment. When GnRHa treatment was initiated at 12, 24, or 36 h during the 2-day period of FSH treatment, inhibition of both FSH- and LH-stimulated adenylate cyclase was observed in ovaries collected at 48 h. Whereas FSH treatment increased the ovarian FSH receptor concentration by more than 100%, concomitant treatment with GnRHa prevented this increase and reduced FSH receptors to 60% of the control level. Treatment with GnRHa alone caused a 65% decrease in FSH receptor levels below the untreated control values. Histological analysis of hormone-treated ovaries indicated that FSH stimulated follicle growth and antrum formation, but caused little luteinization. GnRHa did not completely prevent the effects of FSH on follicle growth, but did induce degeneration and premature cleavage of ova. GnRHa alone suppressed the diethylstilbestrol-stimulated mitotic activity, slightly increased degenerative changes in granulosa cells, and caused oocyte cleavage and premature antrum formation. These findings demonstrate that GnRHa inhibits FSH-dependent adenylate cyclase by a mechanism involving the loss of binding sites for FSH. It is also evident that only short term exposure to GnRHa is necessary for expression of the inhibitory action of the peptide upon FSH- and LH-stimulated adenylate cyclase. PMID- 6295749 TI - Receptors for insulin-like growth factors and growth effects of multiplication stimulating activity (rat insulin-like growth factor II) in rat embryo fibroblasts. AB - Levels of multiplication-stimulating activity (MSA) in fetal rat serum are high (2-4 micrograms/ml), suggesting that MSA may have a role in fetal growth. We now demonstrate that fibroblasts derived from rat embryos (REFs) have specific MSA receptors and respond to MSA with increased DNA synthesis. Two types of insulin like growth factor (IGF) receptors were demonstrated by competitive binding of radioiodinated MSA, IGF-I, and IGF-II and by chemical cross-linking of [125I]iodo MSA and [125I]iodo-IGF-I to REFs. One type of receptor (mol wt, 260,000 under reducing conditions) did not interact with insulin, and another type of receptor (mol wt, 130,000, under reducing conditions) was recognized by insulin. Scatchard analysis of [125I]iodo-MSA binding data was consistent with one class of noninteracting binding sites. A biological response of MSA, increased DNA synthesis, was demonstrated with autoradiography in REFs. During a 16-hr incubation, DNA synthesis was stimulated by normal rat serum, and platelet-poor plasma plus platelet-derived growth factor (PDGF), but not by serum from hypophysectomized (hypox) rats or hypophysectomized (hypox) platelet-poor plasma plus PDGF. However, when MSA was added to hypox serum or to hypox platelet-poor plasma plus PDGF, DNA synthesis was stimulated to the level achieved by normal rat serum. By contrast, during a longer cell multiplication experiment, REFs grew equally well in normal or hypox rat serum, raising the possibility that REFs may produce a MSA-like factor. PMID- 6295750 TI - Insulin receptors in virus-induced diabetes mellitus in mice. AB - Quantitative estimations were made of insulin receptors on liver cell membrane of DBA/2 mice infected with M variant of encephalomyocarditis virus. The virus produced an impairment of glucose metabolism on day 3 of infection, which lasted for 5 months. The fasting plasma insulin concentration was markedly decreased on day 14. The specific binding of 125-I insulin to the membrane receptor was significantly decreased on day 3 of infection. The binding inhibitions were stronger in male mice than in females. The number of insulin receptors began to decrease on day 1, was decreased remarkably by day 3, and returned on day 7 to the level before infection. A decrease of receptor affinity was also observed in infected animals. These results seem to show that changes in insulin receptors are one cause of the impairment of glucose metabolism in the initial phase of virus-induced diabetes. PMID- 6295751 TI - Altered cyclic AMP responsiveness to vasopressin in rat renal medullary dispersed cells by acute elevation of endogenous vasopressin. AB - Medullary and cortical tubular cells were prepared from rat kidneys with collagenase treatment. Arginine vasopressin (AVP) stimulated cyclic AMP production both in medullary and cortical cells with a dose-response relationship at concentrations ranging from 10 microU/ml to 10 mU/ml, whereas parathyroid hormone (PTH) and calcitonin did only in the latter. Using this medullary cell system, effects of acute changes in endogenous plasma AVP levels in vivo on its cyclic AMP responsiveness to AVP (10 mU/ml) in vitro were examined. Acute elevation of plasma AVP levels induced by ip injection of 20% (w/v) polyethylene glycol-isotonic saline solution 3 h prior to sacrifice resulted in a 33% decrease in cyclic AMP responsiveness to AVP (desensitization). More prolonged elevation of plasma AVP levels by water restriction for 48 h, on the other hand, increased the responsiveness by 38 to 81%, which was restored to basal levels by ad libitum intake of water for another 48 h (positive feedback regulation). These maneuvers did not alter the cyclic AMP responsiveness to PTH (10 micrograms/ml) in cortical cells. The results suggest that AVP-stimulated adenyl cyclase in rat renal medulla may be regulated by changes in endogenous AVP levels even within the physiological range. PMID- 6295752 TI - The vitamin D endocrine system: steroid metabolism, hormone receptors, and biological response (calcium binding proteins). PMID- 6295753 TI - Regulation of the seminiferous epithelium. PMID- 6295754 TI - Nucleoside diphosphatase: membrane asymmetry, membrane binding and latency of a peripheral membrane protein. AB - Nucleoside diphosphatase is a peripheral protein of the endoplasmic reticulum in the liver. This review brings together the available information on the properties of the membrane-associated form of the enzyme found in microsomal vesicles. The data are consistent with the view that the enzyme is loosely bound to the inner surface of the vesicles and, therefore, to the luminal surface of the endoplasmic reticulum in vivo. PMID- 6295755 TI - alpha-Adrenergic stimulation of glycolysis and Na+, K+-transport in perfused rat liver. AB - Interactions between alpha-adrenergic-induced lactate production, respiration and K+ uptake of perfused rat livers were investigated. Inhibition of mitochondrial electron transport by cyanide had no effect on K+ uptake, decreased phenylephrine induced lactate production by 60% and suppressed extra O2 consumption by more than 90%. Ouabain did not affect alpha-adrenergic-induced respiration but inhibited K+ uptake and extra lactate production. Effects of ouabain and K+-free medium on lactate production were additive and independent of cyanide, indicating that they were mediated by extramitochondrial mechanisms. Extra ATP utilized for K+ uptake in response to phenylephrine was apparently of glycolytic rather than mitochondrial origin. We propose that alpha-adrenergic-induced lactate production is secondary to an enhanced operation of the plasma membrane (Na+,K+)-ATPase. PMID- 6295756 TI - Synthesis of the proteins of complex III of the mitochondrial respiratory chain in heme-deficient cells. AB - The presence of several proteins of complex III of the respiratory chain has been demonstrated in mitochondria from a mutant of Saccharomyces cerevisiae lacking 5 aminolevulinic acid synthase and, hence, devoid of heme. The two 'core' proteins, apocytochrome b and the iron-sulfur protein, were observed in equal amounts in the heme-deficient and heme-sufficient cells with antiserum against complex III and the sensitive immuno-transfer technique. In addition, three other bands were detected with the complex III antiserum in the mitochondria from the cells lacking heme. One of these has a molecular weight similar to that reported for a precursor form of cytochrome c1. By contrast, when mitochondria from the heme deficient cells were solubilized with mild detergents and treated with the complex III antiserum, almost no immunoprecipitation was obtained above that obtained with control serum. The presence of only one major labeled band with a molecular weight similar to subunit I was observed after gel electrophoresis. These results suggest that heme may be necessary for proper processing of the apoprotein of cytochrome c1 and for the assembly into the membrane of the subunits of complex III, rather than for the synthesis of the proteins. PMID- 6295757 TI - NMN adenylyltransferase: its association with chromatin and with poly(ADP-ribose) polymerase. AB - The nuclear location of NMN adenylytransferase, which catalyses the formation of NAD and pyrophosphate from ATP and NMN, has been examined to ascertain if the enzyme is bound to the domains of chromatin which undergo poly(ADP-ribos)ylation. This latter reaction utilizes much of the cellular NAD. A radioisotope assay using [alpha-32P]ATP was developed to enable precise measurement of picomole amounts of NAD. With this assay, it appeared that the reaction catalysed by NMN adenylyltransferase proceeded with a rapid, early 'burst' of NAD before steady state velocities were established. From this it was calculated that there could be 10- active sites of NMN adenylyltransferase per HeLa nucleus in asynchronously growing cells: that is, approximately one per 10-20 nucleosomes. Very little enzyme activity was liberated by digesting HeLa nuclei with micrococcal nuclease in 80 mM NaCl, and the enzyme which was solubilized was not bound to oligonucleosomes separated by electrophoresis on polyacrylamide gels. In contrast, poly(ADP-ribose) polymerase activity was clearly demonstrated on these particles. The enzyme was readily liberated by DNase I digestion, especially when the digestion was carried out in low-ionic-strength buffer. The results demonstrated that the enzyme was neither bound to oligonucleosomes nor part of the nuclear envelope or matrix. Preliminary results suggested that there could be some direct channelling of NAD between the two enzymes in intact nuclei. It appears that NMN adenylyltransferase is bound within rather than to chromatin. PMID- 6295758 TI - A genetic approach to reveal the action of the opiate receptor in selected neuroblastoma-glioma cells. Interaction with alpha-adrenoceptors, calmodulin and Ca2+-ATPase. AB - Three clones of neuroblastoma-glioma cells that contain low amounts of calmodulin were selected from the NG108-15 cells after several treatments with high concentrations of chlorpromazine. Purified membranes of the three clones had decreased numbers of both alpha-adrenergic and opiate receptors, monitored with [3H]yohimbine and [3H,D-Ala2]methionine encephalinamide, respectively. No changes were observed in the affinity of these radioactive ligands to the receptors of the selected cells as compared to the parent cells. Addition of bovine brain calmodulin did not affect the binding of [3H,D-Ala2]methionine encephalinamide to the membranes of the selected cells and they had the same number of acetylcholine receptors, determined with 1-quinuclidinyl-[phenyl-4-3H]-benzilate, as the parent NG108-15 cells. The basal ATPase activity in the membranes of the selected cells was 35-50% of the parent cells, with a decreased V value and no significant change in the affinity constant Ka to ATP. Addition of Ca2+ to the purified membranes increased the V of the ATPase in the selected as well as the parent cells but the V of the selected cells remained lower than that of the parent cells. Ca2+ had no effect on the Ka to ATP in either cell type. The Ca2+ dependent ATPase activity of both the parent and the selected cells was also calmodulin-dependent dependent since it was blocked in vitro by chlorpromazine. The co-regulation of opiate and adrenergic receptors and their interaction with calmodulin and Ca2+-ATPase is discussed in view of recent observations indicating biochemical and physiological association between opiates, Ca2+ and adrenergic compounds. PMID- 6295759 TI - Desensitisation of beta-adrenergic responsiveness in vivo. Decreased coupling between receptors and adenylate cyclase in isolated brown-fat cells. AB - Chronic catecholamine stimulation in vivo of brown adipose tissue during acclimation of hamsters to cold does not result in any alteration of beta adrenergic receptor number or affinity when determined in isolated adipocytes by (-)-[3H]dihydroalprenolol binding. Norepinephrine displacement of (-) [3H]dihydroalprenolol showed the same Ki for both groups. However, the slope of the displacement curve was shallower for cells from cold-acclimated animals than for controls. Cyclic AMP accumulation was stimulated by norepinephrine in cells from both groups of animals, although the dose-response curve for cells from cold acclimated animals was shifted to the right and the maximum value obtained was less than half that found in cells from control animals. The slope of the curve was again lower. Other catecholamines stimulated cAMP accumulation with an order of potency in agreement with a response mediated through beta 1-adrenergic receptors. The dose-response curve for norepinephrine-stimulated oxygen consumption was also shifted to the right for cells from cold-acclimated animals, although the maximal respiration was only slightly reduced. The slope factor was again decreased. The results are interpreted in terms of a reduced coupling between the beta-receptor and the metabolic response in isolated brown adipocytes from cold-acclimated animals as a result of chronic catecholamine stimulation in vivo. PMID- 6295760 TI - Transient kinetics of the one-electron transfer reaction between reduced flavocytochrome b2 and oxidized cytochrome c. Evidence for the existence of a protein complex in the reaction. AB - The one-electron transfer reaction from reduced flavocytochrome b2 (fully reduced by three electron equivalents) to ferricytochrome c, both purified from the yeast Hansenula anomala, has been studied using stopped-flow spectrophotometry in the course of a single turnover, for reactants initially mixed in a heme molar ratio equal to one. The cytochrome c reduction proceeded to completion through an apparently first-order process. Depending on the experimental conditions (concentrations and or ionic strength), the reduction is of second-order or first order character. To interpret these kinetic results computer simulation studies have been performed based on a kinetic scheme involving, besides the formation of a complex before the electron transfer step, intramolecular electron transfer steps within flavocytochrome b2 to maintain the concentration of the specific electron donor center, the reduced cytochrome b2. As far as the cytochrome c reduction rate constant, ka, and its variations were concerned the simulated data showed that this complicated scheme could approximate a mechanism which is by far the simplest, involving only the two former steps. Such a scheme accounts firstly for the hyperbolic dependence of the rate of reduction of cytochrome c, ka, upon reductant concentrations which had provided clear evidence for the kinetic existence of a complex in the reaction pathway. At 5 degrees C the rate constant for the electron transfer is 380 s-1 with an activation energy of 13.8kJ mol-1 (3.3 kcal mol-1). Secondly it predicts the observed variations of ka with ionic strength and provides estimates of the rate constants of the binding step. PMID- 6295761 TI - Properties and subcellular localization of acid phosphatase activity in cultured arterial smooth muscle cells. PMID- 6295762 TI - On the enigma of old yellow enzyme's spectral properties. AB - Old yellow enzyme (NADPH oxidoreductase) in the free and complexed state was thoroughly investigated by the following techniques: absorption, circular dichroism, fluorescence/phosphorescence and electron paramagnetic resonance spectroscopy and fluorescence/phosphorescence decay measurements, applied over a wide range of temperature (7-293K). The data obtained were interpreted by comparison with results from similar measurements on free FMN, existing spectral data on isoalloxazine model systems and theoretical data. The results clearly demonstrate the inadequacy of a simple phenolate-FMN donor-acceptor charge transfer complex to explain the phenomena occurring upon the addition of phenols to old yellow enzyme. Instead it was found that the phenolate anion interferes strongly with an existing tight complex between FMN and the apoprotein, probably an H-bonded structure in which FMN is tautomerized and interacts with an L-chiral center. This is concluded from a separate electronic transition with an origin at 496 nm, thus far not recognized as such, and the circular dichroism observed. The emission is dominated by that of free FMN, although protein-bound FMN seems also to become luminescent in glassy solution at 143 K. A second fluorescence/phosphorescence emission appears upon excitation of both native and complexed old yellow enzyme in the ultraviolet. This emission is quenched by the addition of phenol to the enzyme, shows a large (3000-cm-1) blue shift on going to a low-temperature glass and is tentatively assigned to excimers of nucleic acids. Long-wavelength excitation with a synchronously pumped, mode-locked Rhodamine 6-G dye laser revealed a third, extremely weak emission in both native old yellow enzyme and its complexes. It decays with a lifetime of about 3 ns at 143 K. Electron paramagnetic resonance spectra revealed the presence of a low amount of an unpaired spin in old yellow enzyme. Owing to an unusual relaxational behaviour it could only be observed below 15 K and, again, the signal was measured in both the native enzyme and its complexes. Possible assignment and consequences of this observation are discussed. In frozen aqueous solutions of the enzyme-phenolate complex, a phase transition was discovered at which the colour of the complex reverted to that of the native enzyme. Subsequent melting restored the original colour. The observed phenomena and existing literature data lead to the conclusion that the only model from which no apparent inconsistencies emerge is that of a very complicated network of hydrogen-bonded structures in the protein. These involve several, partly unknown, chromophores. Phenols interfere with this network, leading to the formation of the long-wavelength absorption band in old yellow enzyme. PMID- 6295763 TI - Thermodynamics of stacking and of self-association of the dinucleoside monophosphate m2(6)A-U from proton NMR chemical shifts: differential concentration temperature profile method. AB - Chemical shifts of base and sugar protons of the modified ribodinucleoside monophosphate N6-dimethyladenylyl(3'-5')uridine (m2(6)A-U) were measured at 100, 360 and 400 MHz in aqueous solution. Seven different samples were used with concentrations ranging from 0.28 mM to 32.7 mM. The temperature was varied from 5 degrees C to 105 degrees C. An internal temperature calibration was used. The effects of intermolecular self-association and of intramolecular stacking on the chemical shifts were quantitatively separated by means of a new approach: differential concentration/temperature profiles (DCTP). Several computational models were tested and the analysis allowed deeper insight into the behaviour of m2(6)A-U at the molecular level. The simple two-state approach for both self association and stacking already afforded a significant improvement over models in which the association is entirely neglected. A computer least-squares analysis of the chemical shift behaviour of each individual proton yielded thermodynamic parameters for self-association and stacking. However, the two-state model did not suffice to reproduce accurately all of the observations. A satisfactory fit required two additional assumptions: (a) the aromatic protons experience different association shifts in stacked and in unstacked molecules: (b) a temperature-dependent conformational equilibrium exists between sets of unstacked microstates. The stacked state is taken to represent a single conformational species. The implementation of this extended model in the least-squares optimization allowed the reproduction of over one thousand chemical shift observations within experimental error. Thermodynamic equilibrium parameters deduced for intramolecular stacking are: delta H degrees x = -28.8 kJ mol-1, delta S degrees x = -93 J mol-1 K-1. These numbers agree well with those obtained earlier by us from circular dichroism spectra. The equilibrium enthalpy and entropy values deduced for the association process are: delta H degrees A = -35 kJ mol-1 and delta S degrees A = -95 J mol-1 K-1. PMID- 6295764 TI - Partial amino-acid sequence of the epidermal growth-factor-binding protein. AB - The partial amino acid sequence of the epidermal growth-factor-binding protein was determined. Residues in 108 unique positions, corresponding to 45% of the molecule, were identified. The protein is a serine protease, closely related to the nerve growth factor gamma subunit. It is suggested that the epidermal growth factor-binding protein, like other serine protease, is synthesized as a single polypeptide chain which undergoes limited endoproteolysis. The isolated material also contained minor amounts of a second serine protease. This protease is closely related to the epidermal growth-factor-binding protein, differing from it in 7 out of the 45 amino acid positions available for comparison. The latter protease may be identical to the previously described protease A. PMID- 6295765 TI - Fatty acids as acute regulators of the proton conductance of hamster brown-fat mitochondria. AB - Possible mechanisms are evaluated for the acute regulation of the hamster brown fat mitochondrial proton-conductance pathway which is active during non-shivering thermogenesis. Isolated mitochondria are incubated under conditions designed to approximate to the non-thermogenic state, and the effect of the steady infusion of fatty acids or acyl derivatives upon respiration, membrane potential and membrane proton conductance is monitored continuously. Fatty acids increase the proton conductance with no detectable threshold concentration, allowing the generated acyl carnitine to be rapidly oxidized. The extent of depolarization and of respiratory increase is a function of the rate of infusion. Immediately infusion is terminated the conductance decreases, the mitochondria repolarize and respiration returns to the initial rate. Infusion of acyl-CoA and acylcarnitine cause only a slight depolarization or respiratory increase after high concentrations of these derivatives have accumulated. Any factor which decreases the rate of conversion of fatty acid to acyl-CoA potentiates the conductance increase. An effect of acyl-CoA upon chloride permeability is not specific to brown-fat mitochondria. Fatty acids infused into rat liver mitochondrial incubations produced a small conductance increase, comparable to that of acyl-CoA or acylcarnitine. It is concluded that fatty acids are the most plausible acute regulators of the proton conductance. The relation to the brown-fat-specific 32000-Mr protein is discussed. PMID- 6295766 TI - The acute regulation of mitochondrial proton conductance in cells and mitochondria from the brown fat of cold-adapted and warm-adapted guinea pigs. AB - Cells and mitochondria were prepared from the brown adipose tissue of adult guinea-pigs adapted to either 4-7 degrees C or 22-25 degrees C. The cold-adapted cells displayed noradrenaline-stimulated, propranolol-sensitive respiration, but noradrenaline failed to increase the respiration of the warm-adapted cells. Purine-nucleotide-sensitive proton conductance was greater in cold-adapted mitochondria than in warm-adapted controls. At the same time cold-adapted mitochondria were extremely sensitive to the uncoupling effect of endogenous and infused fatty acids, and resembled the mitochondria from the brown adipose tissue of cold-adapted hamsters. Warm-adapted mitochondria were ninefold less sensitive, and resembled liver mitochondria. With cold-adapted, but not warm-adapted mitochondria, respiration increased proportionately to the rate of fatty acid infusion. It is concluded that the presence of the 32000-Mr proton conductance pathway is necessary for the expression of a high sensitivity to fatty acid uncoupling, suggesting that the fatty acids interact directly with this protein to modulate the proton conductance during the acute regulation of thermogenesis. PMID- 6295767 TI - Optimizing renal DMSA-scintigraphy with 7-pinhole collimator. AB - Multiple pinhole emission tomographic imaging techniques are currently being applied to imaging of organs of a limited size. The purpose of this study was to evaluate the feasibility of this technique in kidney imaging with Tc99m-DMSA. A 7 pinhole collimator having 4.5 mm. pinhole apertures was used in conjunction with a widefield camera. Left and right kidney were imaged separately. Twelve small renal cysts with a diameter of 1 to 3.5 cm. verified by US or CAT were investigated by 7-pinhole and planar scintigraphy. Eight of 12 renal cysts were identified by 7-pinhole scintigraphy whereas only one cyst was seen by planar scintigraphy. The borderline of cysts detection was 1.5 cm. in 7-pinhole tomography. Basing on these comparative studies and experiences on many patient investigations it seems reasonable to perform renal scintigraphy with 7-pinhole tomography in a routine manner. PMID- 6295768 TI - Flow kinetics of a nucleoside phosphatase common to endoplasmic reticulum, Golgi apparatus, and plasma membrane of rat liver. AB - Nucleoside mono-, di- and triphosphatase activities of highly purified endoplasmic reticulum (ER), Golgi apparatus, and plasma membrane fractions of rat liver were compared. The highest rates of hydrolysis were always in ER or plasma membrane. Golgi apparatus activity was intermediate between those of ER and plasma membrane. This relationship was true for both freshly isolated fractions and salt-extracted membranes. Detergent solubilization of the membranes, polyacrylamide gel electrophoresis of the solubilized proteins, and localization of the enzyme activities on the gel revealed bands of enzyme activity which had identical mobilities in all three membrane fractions as well as other bands of activity that occurred only in ER and to a lesser degree in the Golgi apparatus. Antibodies raised against one of the phosphatase bands of plasma membrane which was common to all three membrane fractions cross-reacted with the corresponding phosphatase band in ER and Golgi apparatus. The anti-nucleoside phosphatase was utilized in combination with pulse-chase techniques to investigate the flow kinetics of transfer of newly synthesized enzyme among different cell compartments. Label first appeared in nucleoside phosphatase within the ER. Maximum specific activity was observed at about 5 min after injection of label and was followed by rapid loss of label. This was followed by appearance of label in Golgi apparatus 15 to 25 min after injection of label and by subsequent rapid loss of label. Plasma membranes were labeled last with no evidence of either rapid accumulation of label or of rapid turnover. Flow of nucleoside phosphatase from its site of synthesis and insertion into the membrane at the endoplasmic reticulum to the plasma membrane via the Golgi apparatus is indicated but in a manner whereby a significant fraction of the protein may be processed (removed?) from the membrane concomitant with the flow process. PMID- 6295769 TI - Free radicals and activated oxygen. AB - Superoxide anion (0(-2)), hydrogen peroxide (H2O2) and hydroxyl radical (OH.) are products of the biological reduction of 0(2). They are very reactive and poorly tolerated within living systems and enzymes that catalytically scavenge these products have been evolved as defense mechanisms. These include superoxide dismutases (SOD), catalase and peroxidases. Large amounts of O-2 are produced by different enzymatic and non enzymatic biological processes. Large amounts of activated oxygens are produced by phagocytosing cells such as macrophages and polymorphonuclear cells. This production is associated with the bactericidal actions of these cells but it also largely contributes to exacerbate and sustain the inflammation where these cells congregate. The arachidonic acid pathway triggered by the inflammatory stimuli is also a source for these oxidizing radicals. The production of activated oxygens has been associated with the normal aging process but also with various toxic reactions (e.g. the toxicity of the herbicide paraquat, of the ionizing radiations, of certain antibiotics such as streptonigrin, etc. . . .). O-2 induces the depolymerization of hyaluronic acid which lends viscosity and lubricating properties to synovial fluids. SOD possess antiinflammatory properties and a bovine SOD, orgotein, has now been largely investigated by intramuscular and intraarticular injections in the treatment of rheumatic diseases. Various antiinflammatory compounds (e.g. the salicylates) are able either to inhibit the production of these oxygen radicals or to scavenge them which seems of importance for their antiinflammatory properties. Singlet oxygen, another activated oxygen, might also play a role in the inflammatory process. PMID- 6295771 TI - Immunopathogenesis of rheumatoid arthritis. AB - Three distinct groups of phenomena occur in the rheumatoid joint. The synovial lining cells proliferate and hypertrophy; the sublining or deep layer becomes infiltrated by mononuclear cells; and fluid, which is rich in polymorphonuclear cells, collects in the synovial space. As a result, there is a polymorphonuclear inflammatory reaction in the rheumatoid effusion and a chronic inflammatory reaction in the sublining layer of synovial membrane. In spite of their divergent histologic character, these two inflammatory reactions appear to be interrelated immunologically. In this review, we describe these two types of inflammation, trace their interrelationship and consider possibly underlying mechanisms for the inflammation. PMID- 6295770 TI - Aspects of resorption and formation of connective tissue during chronic inflammation in rheumatoid arthritis. AB - Connective tissue destruction is a major characteristic of chronic rheumatoid arthritis (RA). Attempts at repair and fibrosis are also seen. This process is accompanied by local cellular and humoral inflammatory reactions. Production of large amount of collagenase and prostaglandin (PGE2) are demonstrated in vivo and can account for the pathogenesis. Long term cultures of adherent synovial cells from patient with RA produce also large amounts of collagenase and PGE2. Collagenase and PGE2 levels can be stimulated with a soluble factor (MCF), a monokine produced by monocyte-macrophages in culture. MCF production is modulated by cellular elements (T lymphocytes), by humoral elements (Fc fragments of immunoglobulin, immune complexes, antigens, lectins), by elements of the matrix (collagen). MCF appears to belong to the category of the interleukin 1. This factor also affects cell replication, collagen synthesis, hormonal response (to PGE2 and PTH). The monocyte-macrophages in this system appear to be the key between the immune and non-immune systems. Studies of MCF (one of the monocyte macrophage products) will help the understanding of the pathogenesis of chronic inflammation such as RA and the designing and screening of new drugs potentially useful in destructive diseases. PMID- 6295772 TI - Immune response to polyoma tumor cells in mice--III. Stimulation of tumor cell growth in vitro by spleen cells from immunized animals. AB - We have evaluated the stimulation of target cell growth in vitro by spleen cells from mice which were immunized with polyoma-transformed cells and other tumor and non-tumor antigens. Stimulation was particularly seen under conditions of immunization that were suboptimal for the production of specific cytotoxicity. Significant stimulation of polyoma target cell growth was observed by lymphocytes from mice immunized against 10(5) Py 4198 tumor cells. This stimulation of target cell growth was not confined to polyoma-transformed cells only. Cells transformed by SV40, H-MuSV and non-transformed cells like 3T3 and embryo fibroblasts were also stimulated. Immunization of mice with syngeneic embryo fibroblasts also resulted in stimulation of tumor cell growth by the spleen cells from the immunized mice. However, the growth stimulation was less consistent and did not occur in all target cells tested. The specificity of immunostimulation was further studied with the Moloney sarcoma virus (M-MuSV) system; an antigenically distinct tumor system. Spleen cells from M-MuSV tumor-bearing mice stimulated cell growth in vitro not only against MuSV-transformed cells but also with SV40 transformed and polyoma-transformed cells as targets. Significant stimulation of target cell growth was also observed by spleen cells from mice that were immunized against 'non-pertinent' antigens, e.g. sheep red blood cells and allogeneic (C57B 1/6) spleen cells. PMID- 6295773 TI - Effect of catecholamines on sterol synthesis in human mononuclear cells. AB - The effect of adrenaline, noradrenaline, isoproterenol and dibutyryl cyclic AMP on the rate of sterol synthesis was studied in freshly isolated human mononuclear cells. Incubation of cells in a medium containing lipid depleted serum led to a two-fold rise in the rate of sterol synthesis from [14C]acetate. Adrenaline, nor adrenaline and isoproterenol added in a concentration of 2 X 10(-5) mol/l at zero time resulted in an inhibition of 30% of this increase in sterol synthesis after 4 h, but had no effect when the hormones were given at 4 h. This indicates that the catecholamines require some time to exert their inhibitory action on sterol synthesis. The action of catecholamines on the incorporation of [14C]acetate into sterols could be mimicked by dibutyryl cyclic AMP. Propranolol added in increasing concentrations to the incubation medium diminished or totally prevented the inhibitory effect of adrenaline, noradrenaline and isoproterenol on sterol synthesis. The results indicate that cholesterol synthesis in extrahepatic human cells, regulated predominantly by low density lipoprotein derived from plasma, is under additional control of catecholamines which may act via cyclic AMP. PMID- 6295774 TI - In vitro and in vivo down-regulation of human platelet alpha 2-adrenoceptors by clonidine. PMID- 6295775 TI - An index for comparing the inhibitory action of vasodilators. AB - An index for comparing the inhibitory effects of vasodilators was developed to gain insight into their mechanism of action on vascular smooth muscle. Rat aortic strips were bathed in Krebs bicarbonate solution and were initially contracted to a stable tension by either phenylephrine or barium chloride. A vasodilator was then added and the remaining tension was noted; this was repeated for cumulative concentrations of vasodilator. At each concentration of vasodilator, the percent reduction in phenylephrine-induced tension (Phe) was compared to the percent reduction in barium-induced tension (Ba) and was expressed as a ratio (Phe/Ba). This ratio clearly separated verapamil and nifedipine (ratio less than 1), which block calcium influx, from papaverine (ratio = 1) which promotes calcium sequestration regardless of the source of calcium, and from dantrolene (ratio greater than 1) which interferes with intracellular calcium mobilization. This index provides a method for comparing the action of those agents presently classified as non-receptor specific vasodilators which act directly on vascular smooth muscle. PMID- 6295776 TI - Autoradiographic localization of opiate kappa-receptors in the guinea-pig brain. AB - Opiate binding sites of the kappa-subtype were visualized in guinea-pig brain sections using in vitro autoradiography. kappa-Binding sites, defined as [3H](-) bremazocine binding in the presence of high concentrations of [D-Ala2,MePhe4,Gly ol5]enkephalin and [D-Ala2,D-Leu5]enkephalin, were found in the cortical laminae V and VI, hippocampal dentate gyrus, and lateral habenulae. The distribution of kappa-sites in the guinea-pig differs considerably from the distribution of mu- and delta-sites which others have found in the rat. PMID- 6295777 TI - Prenatal exposure to electroconvulsive seizures and phenytoin: development of benzodiazepine binding sites, reflex behaviors and seizure thresholds in exposed offspring. AB - Electroconvulsive seizures administered during the period of fetal benzodiazepine (BZ) binding site proliferation (days 16, 18 and 20 in gestation) significantly decreased the density of cerebral cortical benzodiazepine binding sites in exposed pups up to 21 days postnatal age. Significant decreases in seizure threshold were also found in exposed offspring. However, decreases in BZ sites were not accompanied by significant alterations in RO5-4864-displaceable benzodiazepine binding as seen after in utero exposure to phenytoin (DPH). Neither prenatal treatment affected the ability of GABA to enhance BZ binding in vitro. Prenatal exposure to electroshock delayed the onset of acoustic startle and eye-opening during postnatal development; similar effects were not seen following in utero exposure to DPH. These data suggest that prenatal exposure to seizures and DPH decrease postnatal BZ site proliferation by different mechanisms. PMID- 6295778 TI - Improved assays for the assessment of kappa- and delta-properties of opioid ligands. AB - The highly selective non-equilibrium mu-antagonist beta-funaltrexamine (beta-FNA) produced a maximal 20-fold shift in the IC50 for the mu-agonist morphine on the guinea-pig ileum preparation, whilst producing no significant change in the IC50 for the kappa-agonist ethylketazocine. On preparations pretreated with beta-FNA, the pA2 values for the interaction of morphine and ethylketazocine with naloxone were similar. These values were similar to the pA2 value for the interaction of ethylketazocine and naloxone determined on control tissues, but significantly different from the pA2 value for morphine-naloxone on control tissues, indicating that the agonist actions of morphine on preparations pretreated with high concentrations of beta-FNA are mediated by kappa-, rather than mu-receptor interaction. On the mouse vas deferens preparation, co-incubation with the highly selective delta-agonist Tyr-D-Ser-Gly-Phe-Leu-Thr (DSLET) and the non-selective non-equilibrium opiate antagonist beta-chlornaltrexamine (beta-CNA) resulted in marked inhibition of the agonist actions of morphine but had no effect upon the agonist actions of the delta-agonist leucine-enkephalin. The pA2 values for the interactions of naloxone with leucine-enkephalin and etorphine were unaltered by pretreatment with beta-CNA and DSLET. In similarly pretreated tissues, the agonist actions of ethylketazocine were markedly inhibited. It is concluded that manipulation of the guinea-pig ileum and mouse vas deferens preparations in the described manner results in assay systems that possess a largely homogeneous receptor population, and as such are valuable tools with which to evaluate opioid activity. PMID- 6295779 TI - Functional identification of adenylate cyclase-coupled adenosine receptors in rat brain microvessels. AB - It was investigated whether adenosine affects the adenylate cyclase system of microvessels, isolated from the rat cerebral cortex, via an external receptor recently classified as R-site receptor. Several adenosine analogs caused GTP dependent stimulation of adenylate cyclase. The rank order of potency: NECA (5' (N-ethylcarboxamido)-adenosine, EC50 0.2 microM) greater than adenosine (0.71 microM) = 2-chloroadenosine (0.72 microM) greater than L-PIA (N6-(L phenylisopropyl)-adenosine, 1.03 microM) greater than D-PIA (N6-(D phenylisopropyl)-adenosine, 5.27 microM) is consistent with that for agonism at activatory (Ra-site) adenosine receptors in other tissues. Adenylate cyclase stimulation by PIA displayed stereoselectivity. The action of NECA was competitively antagonized by 8-phenyltheophylline. These findings provide functional evidence for Ra-site adenosine receptors in rat brain microvessels. However, direct identification of these receptors by binding studies was not possible. Binding of [3H]NECA to rat brain microvessels displayed rapid on-off kinetics and saturability, but equivocal specificity. PMID- 6295780 TI - Characterization of (-)-[3H]dihydroalprenolol binding to intact and broken cell preparations of human peripheral blood lymphocytes. AB - In this study we compared characteristics of (-)-[3H]dihydroalprenolol ([3H]DHA) binding sites in crude membrane preparations of human peripheral blood lymphocytes with those of intact, viable cells. A valid determination of specific beta-adrenergic receptor binding in both preparations was obtained by defining non-specific [3H]DHA binding with 10(-6) M l- or dl-propranolol or 10(-3) M l isoproterenol. Higher concentrations of propranolol were used in prior reports on lymphocyte membranes. We showed that these concentrations may inhibit non specific binding, causing non-saturability and inhomogeneity of the estimated 'specific' binding. In the intact cell preparations, inclusion of 10(-4) M phentolamine was necessary to reduce the high degree of non-specific binding. By contrast, phentolamine (10(-4) M) showed no effect on the [3H]DHA binding to membrane preparations. At 37 degrees C the [3H]DHA binding to beta-adrenergic receptor sites in both intact and broken cell preparations was rapid and reversible. The sites were stereoselective, as l-propranolol was about two orders of magnitude more potent to inhibit [3H]DHA binding than was the d-isomer. In both preparations, agonists competed for specific binding with a rank order of potency isoproterenol greater than epinephrine greater than norepinephrine, which indicated a beta 2-type of adrenergic receptor. The specific [3H]DHA binding was saturable and Scatchard analysis revealed comparable numbers of homogeneous, non cooperative binding sites (approximately 1250 receptors/cell in the membrane preparations and 1700 receptors/cell in the intact cells). In spite of these similarities the membrane sites showed a lower affinity for the antagonists [3H]DHA and propranolol than did the intact cell sites, whereas their affinity for the agonists was increased. These differences indicate that the membrane system might be less suited to provide physiologically significant information about the beta-adrenergic receptor system. PMID- 6295781 TI - Adrenal involvement in captopril-induced potentiation of morphine analgesia. AB - The involvement of the angiotensin-adrenal system as a possible mechanism in the potentiation of morphine analgesia by the angiotensin-converting enzyme inhibitor, captopril (SQ 14225), was studied in rats. Captopril pretreatment sensitized the animals to the analgesic effects of morphine while angiotensin II exerted an attenuating influence. These effects, however, were not demonstrable in adrenalectomized animals. Although captopril could inhibit the plasma angiotensin-converting enzyme activity, it appeared to have no significant effect on the brain enzyme. It has been suggested that the effects of captopril and angiotensin II on morphine analgesia are mediated indirectly through their effects on adrenal function. PMID- 6295782 TI - Bicuculline-sensitive GABA receptors in rat ovary. AB - A specific [3H]GABA binding site, characterized by a kinetic constant of 52 nM and by a maximal binding capacity of 17 fmol/mg protein was identified in a membrane preparation of rat ovary. [3H]GABA binding was displaced by muscimol, unlabelled GABA or bicuculline. The accumulation of cyclic AMP in the slices of rat ovary was strongly increased by GABA or muscimol and these effects were antagonized by picrotoxin or bicuculline. Our data suggest a possible role of GABA receptors in the regulation of ovarian function in the rat. PMID- 6295783 TI - Possible presynaptic inhibitory effect of etorphine on sympathetic nerve terminals of guinea-pig heart. AB - Etorphine (1-4 microM) dose dependently reduced the sympathetic response induced by trains of field pulses in guinea-pig isolated atria stimulated at 4 Hz; this effect was antagonized by 10 microM naloxone. Since etorphine did not modify the dose-inotropic effect curve of exogenous noradrenaline in the same preparation, it is suggested that the depressant effect of the opioid agonist was due to stimulation of presynaptic inhibitory opiate receptors on adrenergic nerve terminals of the heart. PMID- 6295784 TI - Comparison of the effects of arylazido aminopropionyl ATP (ANAPP3), an ATP antagonist, on responses of the smooth muscle of the guinea-pig vas deferens to ATP and related nucleotides. PMID- 6295785 TI - Kinetic and pharmacological characterization of vasopressin membrane receptors from human kidney medulla: relation to adenylate cyclase activation. PMID- 6295786 TI - Characterization of [3H]nifedipine binding sites in rabbit myocardium. AB - A specific, high affinity (KD 1.8 nM) binding site for the calcium entry blocking drug [3H]nifedipine was identified in homogenates of rabbit myocardium. [3H]Nifedipine binding was rapid (t1/2 3 min) and reversible (t1/2 11 min). Calcium entry blockers with different chemical structures competed with [3H]nifedipine binding in the potency order: nifedipine much greater than D600 = verapamil greater than tiapamil greater than cinnarizine = prenylamine. Diltiazem and perhexiline did not significantly inhibit [3H]nifedipine binding. The potencies of these drugs to inhibit binding were similar to their abilities to depress contractions of the isolated rabbit papillary muscle. The stereoselectivity of D600 and verapamil ((-)-much greater than (+)-isomers) as inhibitors of papillary muscle contractions was not apparent in [3H]nifedipine competition experiments. The slopes of the concentration-inhibition curves for D600 and verapamil were significantly less than for nifedipine. It is concluded that [3H]nifedipine may be labelling part of the myocardial Ca2+ channel, and that verapamil-like substances and nifedipine differ in their mode of interaction with this binding site. PMID- 6295787 TI - Potentiation by dazoxiben, a thromboxane synthetase inhibitor, of platelet aggregation inhibitory activity of a thromboxane receptor antagonist and of prostacyclin. AB - Dazoxiben, a thromboxane (Tx)-synthetase inhibitor, completely prevented platelet TxB2 synthesis, but not the platelet aggregation induced by arachidonic acid (AA) or ADP. It was also ineffective against platelet aggregation brought about by a prostaglandin (PG) endoperoxide analogue, which does not trigger platelet TxA2 synthesis. The association of dazoxiben with a Tx receptor antagonist or with PGI2 resulted in marked potentiation of the latter compounds as inhibitors of platelet aggregation induced by AA or ADP (second wave), but not by U-46619 (a stable analogue of prostaglandin endoperoxides). It therefore appears that inhibition of Tx synthesis does not modify the platelet response to aggregating stimuli but renders platelets more susceptible to inhibition induced by other compounds. PMID- 6295788 TI - Interaction of 2-phenylaminoadenosine (CV 1808) with adenosine systems in rat tissues. AB - 2-Chloroadenosine (2-CADO) and 2-phenylaminoadenosine (CV 1808) were compared in a CNS purinergic receptor binding assay and the inhibition of neurogenic contractions of the vas deferens. Both 2-CADO and CV 1808 are more potent than adenosine in both preparations. CV 1808 was 10 times more active than dipyridamole in enhancing the response of the vas deferens to exogenous adenosine. Thus, CV 1808 may owe its potent coronary vasodilator activity to both a direct action on adenosine receptors and the ability to augment adenosine responses. PMID- 6295789 TI - TRH receptor binding in retina and pituitary: major species variation. AB - Retinas and anterior pituitary glands of nine readily available mammalian and one avian species have been examined for their TRH-sensitive binding of [3H]-[3-Me His2]TRH. Among mammals, major species variations in TRH receptor binding have been detected in both tissues, amounting to about 100-fold in the retina and 20 fold in the pituitary. In the retina, TRH receptor binding was very high in the rat, quite high in sheep and guinea pig, intermediate in rabbit, pig, and mouse, and low but detectable in chicken, cat, calf and dog. In the pituitary gland, binding was very high in sheep, quite high in rabbit, rat, pig, calf and guinea pig, intermediate in chicken, and fairly low in mouse, cat, and dog. A number of possible interfering variables, including affinity differences, albino vs. non albino strains, barbiturate anesthesia, time after death, sex and estrous cycle, age, and history of light exposure, were considered and/or tested directly, with generally negative results. PMID- 6295790 TI - Retinal degeneration in the dog. III. Abnormal cyclic nucleotide metabolism in rod-cone dysplasia. PMID- 6295791 TI - Role of the oxidative metabolic burst in the antibody-dependent cellular cytotoxicity mediated by neutrophil polymorphonuclears. AB - The purpose of the present study was to investigate the mechanisms by which neutrophil polymorphonuclears (PMN) mediate antibody-dependent cellular cytotoxicity (ADCC). Under experimental conditions which allow target cell phagocytosis, PMN were found to efficiently kill IgG-sensitized ox erythrocytes, as determined by the 51Cr release assay. Inhibition of the target cell ingestion by colchicine did not affect the PMN cytotoxic activity, suggesting that target cell phagocytosis does not represent an essential step in the PMN-mediated ADCC against erythrocytes. PMN from patients with Chronic Granulomatous Disease, who have defective oxidative metabolic burst, displayed an impaired ADCC activity, which was unaffected by changes in the phagocytic capacities induced by colchicine. The results indicate that, under the experimental conditions employed, both the intracellular and the extracellular target cell destruction by PMN involve oxygen-dependent mechanisms. PMID- 6295792 TI - Motor command in the ventrolateral thalamic nucleus: neuronal variability can be overcome by ensemble average. AB - The variability and stationarity of single unit activity and reaction times in a simple reaction time task were investigated by means of unit recordings from the ventrolateral thalamic nucleus in the cat. The study was carried out on eight neurons recorded over many trials and displaying an increase in activity correlated with the performed reaction time. The stationarity of the correlation appeared to be better than that of the unit activity or reaction time taken alone. In this stationary process the neuronal variability is considerable, and the investigator has to use time averages over many trials to be able to extract information from the single unit data. Since the nervous system does not required repeated presentations of a stimulus to trigger a movement, it must be using parallel processing of information throught ensemble averages. A simulation of an ensemble average based upon the data from these eight neurons showed that such a process can be efficient. An evaluation of the number of neurons required to obtain a quasi deterministic neuronal command of the reaction time value, gave the speculative figure of 70. This figure could represent the dimension of the set of neurons with the same characteristics involved in the motor command. PMID- 6295793 TI - [Pontine deglutition neurons in sheep. I. Activity and localization]. PMID- 6295794 TI - Physiological characteristics of the solitario-parabrachial relay neurons with tongue afferent inputs in rats. AB - Among 180 units in the solitary tract nucleus (NTS) of rats, 34 solitario parabrachial relay neurons (SP neurons), were identified by antidromic activation from the parabrachial nucleus. The SP neurons were classified into two groups, fast and slow, according to their antidromic latencies. The responsiveness of the SP in comparison with non-SP neurons was studied by electrical stimulation of three tongue nerves: the lingual (L), chorda tympani (CT) and glossopharyngeal (G) nerves. About half the SP neurons produced a single spike with an orthodromic latency of 2-5 ms, while about one third of them discharged more than two spikes. A few neurons gave rise to a long-lasting discharge consisting of five or six spikes. Some SP neurons were excited by stimulation of the tongue afferents with a low stimulus intensity, but other SP neurons produced spikes at only very high voltages. Fast SP cells were not differentiated from slow SP cells, except that latency of orthodromic responses to CT stimulation was significantly shorter in the former than in the latter (P less than 0.05, Mann-Whitney U-test). Locations of the SP and non-SP neurons, reconstructed histologically, indicate that they do not distribute evenly throughout the mediolateral extent at the rostral pole of the NTS, but clustered in its medial half. PMID- 6295796 TI - Impulse coding of ramp currents intracellularly injected into pyramidal tract neurones. AB - Relationships between the repetitive discharge and dynamic aspects of the input were analyzed in pyramidal tract neurones of the cat. Inputs were intracellularly injected currents reaching a steady level after ramps of different slopes (from 0.02 to 1.5 nA ms-1). Output was the instantaneous frequency of the discharge. During the transient phase, instantaneous frequency appeared to be related both to the velocity of rise and to the intensity of the stimulating current. The dynamic component of the cell response was estimated by subtraction of the intensity-bound component (derived from the steady-state response to current steps). After subtraction, the instantaneous frequency of interspike intervals following the first one became proportional to the ramp slope. The instantaneous frequency of the first interval also increased with the current slope, but at a lower rate than the frequency of other intervals. Moreover its dynamic component virtually stopped growing when the ramp slope exceeded 0.3-0.5 nA ms-1. PMID- 6295795 TI - The dual effects of GABA and related amino acids on the electrical threshold of ventral horn group Ia afferent terminations in the cat. AB - Amino acids were administered microelectrophoretically near the unmyelinated terminations of extensor muscle Ia afferent terminations stimulated electrically in the vicinity of lumbar motoneurones in anaesthetized cats. The predominant effect of one group (structurally related to GABA, poor substrates for in vitro amino acid uptake systems) was a reduction in the threshold (depolarization). The second group (including GABA and structural analogues which are substrates for GABA transport systems in vitro) had biphasic effects, an initial reduction being followed by an increase in threshold. The third group (structurally unrelated to GABA, substrates for amino acid uptake systems) only increased Ia termination thresholds. Reductions in termination thresholds, but not increases, were associated with diminution of synaptically evoked primary afferent depolarization, and were decreased by bicuculline methochloride. Many amino acids increased the electrical resistance of the intraspinal medium near the orifices of the barrels of seven barrel micropipettes, and for L-histidine, one of the third group of amino acids, both this effect and the increased threshold of terminations were reversibly modified by microelectrophoretic ouabain. These observations suggest that GABA-mimetics depolarize Ia terminations by interacting with bicuculline-sensitive receptors similar to those at hyperpolarizing gabergic synapses upon spinal interneurones. In addition, under the experimental conditions used, these and other amino acids increase termination thresholds, probably in the absence of any change in membrane conductance, an effect resulting from alterations in the ionic constitution of the extracellular medium around the orifices of micropipettes ejecting amino acids consequent upon the ouabain-sensitive co-transport of amino acids and sodium ions into neurones and glia. PMID- 6295797 TI - Combined morphological and electrophysiological description of connections between single primary afferent fibres and individual motoneurons in the frog spinal cord. AB - In experiments on the isolated frog spinal cord the relationship between the statistical properties of the unitary EPSP and the number of synaptic contacts was determined when the primary afferent fibre used in evoking the EPSP and the motoneuron in which it was recorded were both stained with HRP. The size of the chemical component of the EPSP corresponds to the number of presynaptic boutons. Less obvious numerical correlation exists between the number of contact zones and the number of binomial units. PMID- 6295798 TI - Neuronal responses to 5-hydroxytryptamine and dorsal raphe stimulation within the globus pallidus of the rat. AB - The projection from the dorsal raphe nucleus (DRN) to the globus pallidus (GP) was investigated electrophysiologically, in the urethane-anesthetized rat together with the responsiveness of cells in the GP to 5-hydroxytryptamine (5-HT) and noradrenaline (NA). The majority of spontaneously active cells in the GP had high regular firing rates. They were unaffected by both DRN stimulation (69/83 cells) and iontophoretically applied 5-HT (38/63 units) or NA (30/42 units) but were inhibited by GABA. A few cells (N = 10) were recorded from, that were spontaneously active but with a much lower and less regular firing rate, which, however, seemed to be much more responsive to 5-HT. In addition, DL-homocysteic acid (DLH) was used to activate silent cells and all seven cells activated in this manner were inhibited by 5-HT. In addition 5/6 cells that had their firings maintained by DLH were inhibited by stimulation of the dorsal raphe. The results show a lack of responsiveness to both 5-HT and DRN stimulation of the typically regular spontaneously active pallidal neurons. There seems to be a small population of normally quiescent cells, however, that is sensitive to 5-HT and receives an input from the DRN. PMID- 6295799 TI - Studies on the cerebellocerebral and thalamocortical projections in squirrel monkeys (Saimiri sciureus). AB - Laminar field potential analysis of the cerebellocerebral and thalamocortical (T C) responses was carried out in the cerebral cortex of squirrel monkeys (Saimiri sciureus) under pentobarbital anesthesia. Anatomical studies using horseradish peroxidase (HRP) were combined with electrophysiological studies. Stimulation of the fastigial nucleus induced bilaterally surface positive-depth negative potentials, i.e., deep T-C responses, in the medial part of the motor cortex and in the parietal cortex (mainly area 5) with a latency of 4 to 5 ms. Stimulation of the interpositus and dentate nucleus evoked contralaterally surface negative depth positive potentials (superficial T-C responses) in the intermediate and lateral part of the motor cortex and in the premotor cortex (area 6) with a latency of 3 to 4 ms. Stimulation of the thalamic nuclei as well as the HRP study revealed that the medial part of V.o.p. (posterior basal part of VL) and Z.o (upper part of VL) and the intralaminar nucleus (CL) mediate the interpositus- and dentate-induced responses to the motor and premotor cortex, and that the ventrolateral part of V.o.p. and Z.im (upper part of VPLo) relays the fastigial induced responses to the motor cortex. PMID- 6295800 TI - Poor myelination in the central nervous system of "dilute-lethal mutant mice" (d1/d1). AB - The activity of 2',3'-cyclic nucleotide 3'-phosphohydrolase (CNPase), as a marker for myelinogenesis, was measured in different parts of the nervous system of dilute-lethal mutant mice (d1/d1). The activity in terms of micromoles of 2'-AMP formed per milligram of protein of homogenate per minute was found to be exceedingly reduced in the cerebrum, brain stem, and medulla oblongata, slightly reduced in the cerebellum and cervical spinal cord, but not reduced in the thoracic spinal cord and the optic and sciatic nerves. These results clearly indicate that dilute-lethal mutant mice show poor myelination in the cerebrum, brain stem, and medulla oblongata. PMID- 6295801 TI - Long-lasting facilitation of a monosynaptic pathway as a result of "partial" axotomy of its presynaptic elements. AB - We investigated the long-lasting effects of severing an axonal branch on the function of the synapses made by the remaining branches of the "partially" transected axon. The experimental model used was the monosynaptic reflex pathway of the cat lumbar cord. Preliminary surgical procedures consisted of a dorsal column (DC) hemisection at the L3--L4 level on the left side under aseptic conditions. During the final recording experiments, the monosynaptic reflex (MSR) of the operated and normal (control) side was studied in motorpools in the L7 and S1 segments. The results indicate that the DC operation produced on the operated side (a) an enhancement of the MSR and (b) a more powerful postetanic potentiation, both of the "early" and "late" varieties. These effects were seen as early as 34 days and as late as 121 days after the original DC hemisection. Control experiments were carried out to exclude denervation supersensitivity, supraspinal effects, and normal asymmetries between sides as the mechanism underlying these effects. A presynaptic change in the operated axons, resulting in an increased synaptic efficiency, is proposed as the possible mechanism. PMID- 6295802 TI - Entamoeba histolytica: correlation between virulence and content of proteolytic enzymes. AB - Intact trophozoites of the virulent Entamoeba histolytica strain HM-1:IMSS (HM-1) destroyed a monolayer of baby hamster kidney (BHK) cells at a higher rate and efficiency than trophozoites of the nonvirulent strain HK-9. The destructive effect could be partially attributed to the proteolytic activity of the amoeba, since quantitative differences were found in the enzymatic activity of the two strains tested. Crude extracts or secreted enzymes of HM-1 trophozoites digested Azocoll, as well as the bovine cold-insoluble globulin fraction, at a much higher rate than the corresponding preparations from HK-9. This proteolytic activity was found to be activated by free sulfhydryl groups. Sodium dodecyl sulfate polyacrylamide gel electrophoresis of the BHK cell proteins of pre- and postamoebic activities showed patterns similar to the trypsin effect on the same target cells. These enzymes were found to digest the proteins participating in the attachment of the target cells to the substrate and, consequently, cause detachment of these cells. PMID- 6295803 TI - Neutralization of the fibrogenic silica-released macrophage factor by antiserum. AB - Antiserum against the fibrogenic factor from the culture medium of silica-treated rat macrophages was produced in rabbits. In dilution 1: 10,000 the antiserum cancelled the stimulating effect of the culture medium of rat macrophages on the 3H-proline incorporation into granulation-tissue fibroblasts. The rabbit antiserum had variable effects on the stimulation by the media of silica-treated human monocytes, but no effect on the media of cultured human SU-DHL-2-cells. PMID- 6295804 TI - Vascular smooth muscle: intracellular aspects of adrenergic receptor contraction coupling. PMID- 6295805 TI - Effect of hyperphenylalaninemia on polyphosphoinositides content of rat brain. PMID- 6295806 TI - [Effect of corticotropin (porcine) on adrenal function in burn injury patients]. AB - In the blood of patients with burns comprising 20-49% of the body surface, the hydrocortisone level was 2-3 times as increased as compared with healthy subjects during 4 weeks after the injury. Administration of high-purified porcine corticotropin significantly raised the hydrocortisone level by 2 times. The maximal absolute increase in the hydrocortisone content in the blood was provoked by corticotropin in the patients within the first week, whereas the minimal increase within the 3rd week of the burn disease. Repeated administration of porcine corticotropin produced no allergic reactions, anaphylactic manifestations, local or general complications. PMID- 6295808 TI - A sharp structural transition in pA03 plasmid DNA caused by increased superhelix density. PMID- 6295807 TI - Specific inhibition of benzodiazepine receptor binding by some 6H-indolo[2,3-b] [1,8]naphthyridines and 11H-indolo [3,2-c] [1,8]naphthyridines. AB - The reactivity of some 6H-indolo[2,3-b] [1,8]naphthyridines (II) and 11H indolo[3,2-c] [1,8]naphthyridines (III) in displacing specific [3H] diazepam binding from bovine brain membranes was examined. All the indolonaphthyridines tested are active and show a higher activity than indole and tryptophan. The inhibition is due to direct interaction with the benzodiazepine binding sites. Some structure-activity relationships are discussed. PMID- 6295809 TI - Potentiating effect of calmodulin and catalytic subunit of cyclic AMP-dependent protein kinase on ATP-dependent Ca2+-transport by cardiac sarcolemma. AB - Highly purified sarcolemmal membranes were prepared from pig heart homogenates by differential and density gradient centrifugations. The membrane fragments exhibit ATP-dependent Ca2+-transport and Na+/Ca2+-exchange activities. ATP-dependent Ca2+ transport (K0.5Ca2+ = 0.3 microM; Vmax = 4.6 nmol Ca2+.mg protein-1.min-1) is not stimulated by oxalate. Ca2+-uptake is also not supported by p-nitro phenylphosphate. Preincubation of sarcolemma with MgATP, calmodulin and catalytic subunit of cyclic AMP-dependent protein kinase stimulates active Ca2+-transport 1.8-fold. The effects of calmodulin and catalytic subunit are potentiating rather than additive. A large portion of the Ca2+ additionally accumulated after prephosphorylation of membranes is exchangeable for Na+ via the Na+/Ca2+-exchange system. PMID- 6295810 TI - Identification of putative calcium channels in skeletal muscle microsomes. AB - Saturable binding sites for the labelled calcium antagonist (+/-)[3H]nimodipine were found in guinea-pig hind limb skeletal muscle homogenates. Binding sites were enriched in a microsomal pellet by differential centrifugation of the homogenate. [3H]Nimodipine binding (Kd = 1.5 +/- 0.03 nM, Bmax = 2.1 +/- 0.25 pmol/protein, at 37 degrees C) copurified (6-fold) in this fraction with [3H]ouabain binding (6.6-fold) and 125I-alpha-bungarotoxin binding (5-fold). d cis-Diltiazem (but not 1-cis-diltiazem) stimulated (+/-) [3H]nimodipine binding (ED50 1 microM) by increasing the Bmax. Binding sites discriminated between the optical enantiomers of 1.4-dihydropyridine calcium antagonists and the optically pure enantiomers of D-600. The data confirm, with biochemical techniques, the presence of 1,4-dihydropyridine and (+/-) D-600 inhibitable calcium channels in skeletal muscle, previously found with electrophysiological techniques. PMID- 6295811 TI - Activation of phosphodiesterase by chicken iodopsin. AB - Activation of guanosine 3',5'-cyclic monophosphate phosphodiesterase in outer segment membrane of chicken retina was investigated. Irradiation of dark-adapted chicken outer segment membrane for bleaching of iodopsin increased the enzyme activity twice as much as that in the dark in the presence of GTP. Further irradiation of the sample for bleaching of rhodopsin in the membrane induced some additional activation of the enzyme. However, chicken iodopsin activated the enzyme in frog rod outer segment membrane without irradiation, while chicken rhodopsin did not. Irradiation of chicken iodopsin increased the enzyme activity twice as much as that in the dark. PMID- 6295812 TI - A plasmid region encoding the active fragment and the inhibitor protein of colicin E3--CA38. PMID- 6295813 TI - Purification, Mr-value and subunit structure of a teichoic acid hydrolase from Bacillus subtilis. AB - A teichoic acid degrading enzyme (teichoicase) was purified to apparent homogeneity from a water-soluble cell extract of sporulating Bacillus subtilis cells. A rapid test for the detection of teichoicase activity was developed. The purified teichoicase has an app. Mr = 310 000. It consists of 4 identical subunits of Mr = 78 000 each. PMID- 6295814 TI - Histamine interaction on surface recognition sites of H2-type in parietal and non parietal cells isolated from the guinea pig stomach. AB - In gastric cells isolated by pronase digestion from the guinea pig, histamine stimulated cAMP production in 3 fundic cell fractions (EC50 = 1.6--2 x 10(-4) M) enriched in parietal (94%), peptic (63%) and mucous cells (87%) as well as in antral cells (EC50 = 4 x 10(-4) M) that are devoid of parietal cells. Histamine stimulations were completely inhibited by the H2 antagonist cimetidine (Ki = 0.27 -0.57 x 10(-6) M) or by the H1 antagonist diphenhydramine, but at 100-times lower potency (Ki = 22--45.7 x 10(-6) M), indicating the presence of histamine H2 receptors in parietal and nonparietal cells of the guinea pig gastric mucosa. PMID- 6295815 TI - An extended Ca2+-hypothesis of visual transduction with a role for cyclic GMP. AB - A model is described having the following features: Light induces Ca2+ release from vertebrate rod outer segments discs via pores composed of multimeric rhodopsin. Cytoplasmic Ca2+ reversibly blocks Na+ channels of the surface membrane, with the time course of development and amplitude of the response to light being influenced by restrictions on intradiscal Ca2+ diffusion. The falling phase of response reflects a decline in cytoplasmic [Ca2+] due to a Ca2+-binding protein controlled by cyclic GMP so that its binding capacity is increased by the reduction in cytoplasmic [cyclic GMP] which follows rhodopsin bleaching. PMID- 6295816 TI - Transmembrane potential variations accompanying the PMA-triggered O-2 and H2O2 release by mouse peritoneal macrophages. AB - Stimulation by PMA of Streptococci-elicited macrophages induced a transient membrane depolarization preceding the onset of detectable O-2 production. Mice resident peritoneal macrophages were unresponsive to PMA for both activities. The PMA-triggered membrane depolarization seemed to be independent from O-2 production because inhibition of membrane depolarization by EGTA had no effect on rates of O-2 or H2O2 release and rate of antimycin A insensitive O2 uptake by Streptococci-elicited macrophages. The portion of O2 uptake recovered as O-2 was found to be 1/3. The rate of O-2 release was twice the rate of H2O2 production (1.1 nmol H2O2.min-1 X 10(6) macrophages-1). PMID- 6295818 TI - Rapid, transient methylation of four proteins in aggregative amoebae of Dictyostelium discoideum as a response to stimulation with cyclic AMP. AB - In Dictyostelium discoideum, extracellular cAMP induces chemotaxis and cell aggregation. Suspensions of cAMP-sensitive cells are shown to respond to a 10(-6) M cAMP-pulse with increased methylation of 4 proteins with app. Mr 110000, 46000, 28000 and 16000. The Mr 110000 and 28000 proteins show a triphasic response with maxima 15, 60 and 150-180 s after stimulation. The responses of the Mr 46000 and 16000 proteins are monophasic, maxima being reached 3 and 15 s after stimulation, respectively. Optimal responses of methylation are observed over 10(-7) - 10(-6) M cAMP. The methylation reaction may be involved in the processing of the chemotactic signal. PMID- 6295817 TI - The binding of dicyclohexylcarbodiimide to cytochrome b of complex III isolated from yeast mitochondria. PMID- 6295819 TI - Pharmacological properties of the converting enzyme inhibitor, enalapril maleate (MK-421). AB - Enalapril maleate (MK-421), an ethyl ester, is an angiotensin-converting enzyme (ACE) inhibitor from a novel series of substituted N-carboxymethyldipeptides. The parent diacid (MK-422) N-[(S)-1-carboxy-3-phenylpropyl]-L-Ala-L-Pro of MK-421 inhibited hog plasma ACE with an I50 of 1.2 nM. Because deesterification occurs slowly or not at all in vitro, the in vitro I50 for enalapril was 1200 nM. However, both enalapril and MK-422 were potent inhibitors of ACE by the i.v. and oral routes in rats and dogs. In rats with experimental hypertension, enalapril was most potent in those models in which the renin-angiotensin system plays a dominant role (salt restriction, two-kidney Grollman) and in models rendered renin dependent by diuretics, although blood pressure reduction did occur in low or normal renin models such as spontaneously hypertensive rats, in which inhibition of ACE as measured by the blockade of angiotensin I pressor responses bore little temporal relationship to the later fall in blood pressure after enalapril. PMID- 6295820 TI - Captopril and MK-421: stability on storage, distribution to the central nervous system, and onset of activity. PMID- 6295821 TI - Antihypertensive properties of ketanserin (R 41 468). AB - The serotonergic receptor antagonist 3-(2-[4-(4-fluorobenzoyl)-1 piperidinyl]ethyl)-2,4-[1H, 3H] quinazolinedione (ketanserin) causes dose dependent inhibition of the effects of 5-hydroxytryptamine (5-HT) on 5-HT2 serotonergic receptors. These receptors mediate facilitation of platelet aggregation, direct vasoconstriction in several arteries and veins, and direct amplification of vasoconstrictor responses to other neurohumoral mediators. Ketanserin does not inhibit vasodilator effects of 5-HT. At higher concentrations, ketanserin has alpha 1-adrenergic blocking properties. The compound causes dose-related reductions in arterial blood pressure in hypertensive animals and humans that are larger and occur at lower doses than in normotensive controls. In humans, the antihypertensive properties of ketanserin do not appear to involve alpha 1-adrenergic inhibition, because the compounds given i.v. (10 mg) do not affect the pressor dose-response curve to phenylephrine. PMID- 6295822 TI - Hypertension and new antihypertensive drugs: clinical perspectives. AB - The nature of the clinical problem posed by hypertension has been changed fundamentally by 30 years of successful drug therapy. Cardiovascular pathology directly related to the high pressure is no longer the main problem. Complications of atheroma, particularly in the coronary arteries, are the main cause of hypertension-related deaths. Most patients now starting treatment have mild hypertension and the benefits, although significant, are small (two deaths prevented per 1000 patient-years of treatment). These two considerations dictate that new drugs must have a very low incidence of toxicity, a low incidence of symptomatic side effects, and preferably, a favorable effect on morbidity and mortality from ischemic heart disease. Several possible approaches have promise: 1) reduction of ischemic heart disease mortality by beta-adrenergic blockade; 2) use of calcium entry blocking drugs, which may combine vasodilation with antiarrhythmic effects; 3) non-thiol-converting enzyme inhibitors, which may have a particularly low incidence of side effects; 4) prostanoid derivatives that may combine vasodilatation with an antiplatelet action. PMID- 6295823 TI - 5-hydroxytryptamine and precapillary vessels. AB - The complexity of the vascular effects of 5-hydroxytryptamine (5-HT) is illustrated by differences in sensitivity to the amine among arterial tissues of different origin. The interaction of 5-HT with 5-HT2 receptors is inhibited by specific antagonists such as ketanserin and methysergide. Such compounds also inhibit the contractile responses to endogenous 5-HT released from aggregating platelets. The vasodilator component of the response to 5-HT can be unmasked in the presence of serotonergic blockade, provided the antagonist used has no partial agonistic properties. 5-HT augments (amplifies) the vasoconstrictor responses to adrenergic and nonadrenergic neurohumoral mediators. The amplifying effect of the monoamine is prevented by 5-HT2-serotonergic antagonists such as ketanserin. PMID- 6295824 TI - Potassium effects on contraction in arterial smooth muscle mediated by Na+, K+ ATPase. AB - A local increase in the extracellular potassium concentration [K+]o, up to about 8 meq/liter either by topical application or intra-arterial infusion of K+ salts, causes arteriolar dilation and decreased resistance to blood flow in systemic vascular beds. Isolated vascular smooth muscle responds to a similar increase in [K+] in the bathing fluid with relaxation if the preparation has some initial active tension. Reduction in [K+] over physiological ranges produces arteriolar constriction and increased resistance to blood flow. K+ vasodilation is accompanied by hyperpolarization of the smooth muscle cell whereas the vasoconstriction is accompanied by depolarization. All these responses can be blocked by ouabain, a potent Na+, K+-ATPase inhibitor. It is therefore thought that K+ vasodilation results from stimulation of the electrogenic Na+-K+ pump whereas the vasoconstriction results from inhibition of this pump. A number of conditions that alter resistance also alter interstitial fluid [K+]. These include exercise, myocardial ischemia, epileptic convulsions, and evoked electrical activity of the somatomotor cortex. Certain findings, including those during administration of ouabain, suggest that changes in [K+] contribute significantly to some of the changes in resistance. PMID- 6295825 TI - Sodium pump hyperpolarization-relaxation in rat caudal artery. AB - Electrogenic ion transport contributes vitally to the Em in vascular muscle and thus is an important influence on contraction and relaxation. Agents that act on membrane ion transport will cause depolarization or hyperpolarization of sufficient magnitude to cause contraction or relaxation, respectively. In the caudal artery of the rat, the principal ion involved appears to be Na+. The transport process appears to be the Na+, K+-ATPase, which is ouabain sensitive, rather than other possible candidates such as the Na+-Ca2+ countertransport mechanism. The hyperpolarization and parallel relaxation found in caudal artery on return to K+ provide unequivocal evidence for an electrogenic Na+ pump. In contrast, the lack of a contraction on transition to O Na+ suggests that the caudal artery does not show an Na+-K+ countertransport system. Although other ion transport systems might be established later for caudal artery and other kinds of vascular muscle, it now appears that the electrogenic Na+ pump is the main ion transport system controlling contraction through a continuous contribution to Em. PMID- 6295826 TI - beta-Adrenergic receptor isolation and characterization with immobilized drugs and monoclonal antibodies. AB - Immobilized catecholamines have played an important role in the localization of alpha- and beta-adrenergic receptors to the plasma membrane of effector cells, and in elucidating mechanisms of beta receptor activation of cardiac muscle. An extension of immobilized drug and affinity chromatography procedures has been developed by utilizing receptor-specific monoclonal antibodies. Structurally different beta 1- and beta 2-adrenergic receptors have been purified with a single monoclonal antibody affinity column, where the antibody is specific for an epitope in the ligand-binding site of both beta 1 and beta 2 receptors. Specificity was increased by elution of receptors from the monoclonal antibody affinity columns with low concentrations of beta-receptor antagonists. These studies indicate that the turkey erythrocyte beta 1-adrenergic receptor is most likely a monomer with a molecular weight of 65,000-70,000. beta 2-Adrenergic receptors have a primary subunit of 55,000-58,000 daltons, with the intact receptor in membranes having a molecular weight of 109,000, which suggests that the beta 2-adrenergic receptor is most likely a dimer of either two identical subunits or a binding subunit and an unidentified second subunit. PMID- 6295827 TI - The effects of prolonged retention of diaphragms on colonization by Staphylococcus aureus of the lower genital tract. AB - To determine whether prolonged retention of diaphragms alters Staphylococcus aureus colonization of the lower genital tract, 183 nonpregnant, asymptomatic women were screened for the presence of this pathogen in the cervix and upper vagina. Nineteen (10.4%) had positive cultures: 5 from the vagina, 8 from the cervix, and 6 from both sites. A second examination with quantitation revealed the presence of S. aureus in 7 of the 19 patients. In five of the six available patients, the number of staphylococci increased significantly after prolonged (24 hour) retention of diaphragms and decreased in one subject only. In two additional patients S. aureus was recovered after, but not before, the use of diaphragms. An increase in the number of other microorganisms also was noted after prolonged retention of diaphragms. In vitro study of the spermicide nonoxynol-9 revealed that it had no inhibitory activity on staphylococci or group B streptococci but was highly effective against Streptococcus pneumoniae. PMID- 6295828 TI - The effects of marijuana extract and delta 9-tetrahydrocannabinol on luteal function in the rhesus monkey. AB - The effects of marijuana extract (ME) and delta 9-tetrahydrocannabinol (THC) on corpus luteum function were studied in the rhesus monkey by the use of in vivo and in vitro techniques. THC (2.5 mg/kg) or vehicle (3% Tween 80 in saline) was administered by an intramuscular injection to rhesus monkeys on day 20, 21, or 22 of the menstrual cycle. Progesterone (P) levels were measured at 6-hour intervals for the first 24 hours after treatment. THC caused a significant decrease in P levels during this 24-hour period. This decrease was reversed by the administration of human chorionic gonadotropin (hCG) at 6 hours after THC administration. When THC was administered 2 hours after hCG, it failed to inhibit the expected rise in serum P levels caused by hCG. Direct effects of the drugs on P production were studied with the use of dispersed luteal cells obtained from monkeys on day 21 or 22 of the menstrual cycle. Neither ME nor THC had any effect on basal P production in these in vitro studies. These data suggest that the inhibitory effect of THC on P levels during the luteal phase are not mediated by a direct effect of the drug on ovarian steroid production. PMID- 6295830 TI - Na+/K+ ATPase and cell growth--III: Enzymatic activity in cultured and EGF stimulated HeLa cells. AB - 1. The behaviour of Na+/K+ ATPase during the growth of ectodermal tumoral cells (HeLa) has been investigated. 2. Besides spontaneously growing cells, also samples stimulated with EGF have been tested. 3. The content of--SH groups in the homogenates of both the untreated and the EGF stimulated samples has been tested. 4. Results show a decrease of enzymatic activity during the culture of neoplastic cells and an enhancement of this behaviour in the EGF stimulated cells due to the action of the hormone on--SH groups. PMID- 6295829 TI - [Hormonal dependence of the initial stages of heparin clearance during immobilization stress in the rat]. AB - The 30-min immobilization stress reduces intensity of initial stages of heparin clearance in rats due to increased secretion of ACTH: in blockade of ACTH release the intensity of heparin clearance is not reduced whereas in intensifying the immobilization stress with ACTH administration it is reduced more than in immobilization alone. The stress does not accelerate heparin clearance in adrenalectomized animals. Substitutional administration of hydrocortisone normalizes the clearance in these animals. This suggests a permissive role of glycocorticoids in regulation of heparin clearance intensity. Detainment of heparin in the blood stream in stress is causally related to activation of anticoagulating system and to increased non-enzymatic fibrinolytie activity of the blood due to intensified formation of complex heparin compounds. PMID- 6295831 TI - Partial purification of protease-solubilised low Km cyclic nucleotide phosphodiesterase from liver. PMID- 6295832 TI - Independent pathways for the anaerobic reduction of nitrite to ammonia by Escherichia coli. PMID- 6295833 TI - The organization of the quinone pool. PMID- 6295834 TI - The reactions of terminal oxidases in Escherichia coli K12 with oxygen and carbon monoxide at sub-zero temperatures. PMID- 6295835 TI - alpha-Adrenoceptors and hypertension. PMID- 6295836 TI - Restoration of beta-adrenergic responsiveness in desensitized cells. PMID- 6295837 TI - alpha and beta-adrenoceptor coupling to adenylate cyclase. PMID- 6295838 TI - Resolution of radioiodinated thyrotropin into receptor active and inactive components by column chromatography. AB - Following radioiodination by the lactoperoxidase method and subsequent purification on Sephadex G100, it was found that [125I]TSH exhibited varying degrees of binding activities to the thyrotropin receptor. In order to further purify the radiolabeled hormone, the [125I]TSH preparation was chromatographed on Sepharose 6B. Two peaks of radioactive material (Peaks I and II) were recovered, containing approx. 60% of the applied radioactivity. Upon elution with Mg2+, the remainder of the radiolabeled material was recovered as a single peak (Peak III). Characterization of these 3 peaks by radioimmunoassay demonstrated that all 3 were immunocompetent, although Peaks I and III were 3-4-fold more immunoreactive than Peak II. Analysis by radioreceptor assay indicated that Peak III showed an increase in receptor-binding capacity (in comparison with the [125I]TSH preparation purified by Sephadex G100 alone), while both Peaks I and II exhibited significantly reduced binding activity. In contrast, human TSH (NIH) chromatographed mainly as a receptor inactive peak, although it was fully immunocompetent. Scatchard analysis of receptor binding to bovine [125I]TSH from Peak III yielded a curvilinear plot with affinities similar to those we have previously reported for [125I]TSH purified by Sephadex G100 chromatography. The total number of binding sites, however, increased proportionally with the active fraction of the [125I]TSH preparation. Since the mass of bound hormone is calculated from the percent bound of total radioactivity and only a fraction of the measured total participates in the binding, it is therefore necessary to correct for the inactive fraction when calculating the total receptor number. PMID- 6295839 TI - Hypotonic activation of insulin-sensitive phosphodiesterase in rat fat cells. AB - The effect of hypotonic treatment on the low Km membrane-bound cyclic AMP phosphodiesterase was investigated. Isolated fat cells obtained from Sprague Dawley rats were incubated at 37 degrees C with an without 2 nM insulin. A crude microsomal fraction prepared by differential centrifugation was suspended in a hypotonic buffer at 4 degrees C, with and without protease inhibitors. Following solubilization from the particulate fraction, hypotonic treatment stimulated the phosphodiesterase in a time-dependent manner. Among the protease inhibitors, E 64, leupeptin and antipain were effective in preventing hypotonic activation of the enzyme. The release of the enzyme from the particulate fraction was partially inhibited by antipain. Kinetic analysis of the enzyme from hypotonic activation was much the same as that of the enzyme from the isotonic buffer. These results suggest that hypotonic activation of the phosphodiesterase may be the result of stimulation of an endogenous thiol protease of lysosomal origin. PMID- 6295840 TI - Changes in pituitary LHRH receptor levels in situations of increased or decreased gonadotrophin secretion in the male rat. PMID- 6295841 TI - Cyclic cytidine 3',5'-monophosphate (cCMP) in cell regulation. PMID- 6295842 TI - Characterization of specific low-density lipoprotein binding sites in human term placental microvillous membranes. AB - Purified microvillous membranes prepared from normal term human placenta were studied for their ability to bind specifically low-density lipoprotein (LDL). Electron microscopic examination of the membrane preparations revealed essentially microvilli-like structures, and the enzyme analyses a 14-17-fold enrichment in the membrane markers 5'-nucleotidase and alkaline phosphatase. The binding of [125I]LDL was dependent on time, temperature, pH and protein concentration; it was saturable with a low capacity (130.4 +/- 22.2 ng/mg of membrane protein) and presented a high affinity (apparent Ka 6.12 +/- 1.32 micrograms protein per ml). These high-affinity binding sites were specific for LDL (high-density lipoprotein induced less competition than unlabelled LDL) and were sensitive to pronase digestion. Unlike the binding of LDL to other tissues, the [125I]LDL binding to microvillous membranes did not require divalent cations. The presence of specific LDL receptors on the placental microvillous membranes, located at the effective site of exchange between the maternal blood and the placental tissue, supports the concept that human placenta utilizes LDL cholesterol for its progesterone synthesis. PMID- 6295843 TI - LH/hCG receptors and stimulation of testosterone biosynthesis in the rat testis: changes during foetal development in vivo and in vitro. AB - The development of gonadotropin receptors to LH/hCG in the foetal rat testis from 14 to 20 days of gestation was monitored by quantitative binding assays using [125I]hCG and compared to testosterone secretion under basal and stimulated conditions in vitro. Specific hCG binding was first detected on day 15. Thereafter the binding increased gradually with advancement in gestational age and correlated with LH-stimulated secretion of testosterone in vitro. On day 18 of gestation the KA was 0.82 x 10(10) M-1 and the binding capacity was 0.57 fmoles per testis. No binding was detectable in the female gonads at this age. The differentiation of hCG receptors obtained in vitro was very low, although it was sufficient to give a full response to LH with testosterone biosynthesis. The results of the present study suggest that functional receptors do not appear before the capacity to synthesize testosterone is expressed and that their appearance is not dependent on factors extrinsic to the testis. However, additional factors could be necessary for a full development of the receptors. PMID- 6295844 TI - Affinities of reactive vasopressin analogues for bovine antidiuretic receptor. PMID- 6295845 TI - Effects of serum and lipoproteins on steroidogenesis in cultured bovine luteal cells. AB - The effects of serum and lipoproteins on the function of bovine luteal cells in tissue culture were examined. Corpora lutea from regularly cycling dairy cows were dissociated with collagenase and cultured in Ham's F-12 medium with or without serum. The serum-free medium was supplemented with insulin, transferrin and hydrocortisone. Addition of LH to the serum-containing medium did not increase progesterone (P4) production. When the luteal cells were cultured in serum-free medium. LH produced a dose-dependent increase (P less than 0.001) in P4 production during the first 24 h of culture. The responsiveness of the cells to LH then gradually declined, and remained low until days 9-11 of culture, at which time the cells regained their ability to respond to LH. The luteal cells were responsive to dibutyryl cAMP in both serum-containing and serum-free medium. Serum lipoproteins (low and high density) were able to produce a 150-260% increase in progesterone production by the luteal cells cultured in serum-free medium. The results indicate that the presence of serum in the cell culture medium inhibits the responsiveness of luteal cells to LH at a step prior to the increase in cellular cAMP, and that serum lipoproteins can be used to increase progesterone production by cultured bovine luteal cells. PMID- 6295846 TI - Control of glycosaminoglycan metabolism by ACTH in bovine adrenocortical cells in primary culture. AB - Bovine adrenocortical cells in primary culture actively incorporated [35S]sodium sulphate and [3H]glucosamine into glycosaminoglycans (GAGs). Most of the synthesized GAGs were found associated with the pericellular material and secreted in the culture medium, while less than 15% remained associated with the cell pellet. In all these fractions the major product was identified by its molecular properties and selective degradation procedures as a heparan sulphate structure. Exposure of the cells to ACTH induces a shift of the synthesized GAGs from the cellular to the pericellular compartment, an increase in the heparan sulphate labelling and the occurrence of an additional product characterized as a hyaluronate. These data suggest that GAG metabolism of the adrenocortical cell may be under hormonal control and open the possibility of a modulation of cell activity through modifications of its GAG components, especially those involved in the cell matrix architecture. PMID- 6295847 TI - Growth hormone receptors in rat liver membranes: effects of fasting and refeeding, and correlation with plasma somatomedin activity. AB - The effects of fasting and refeeding on hepatic growth hormone receptors, on insulin receptors and on plasma somatomedin activity were studied. Female rats were either subjected to fasting for 4 days, refed for 3 days after a 4-day fasting, or allowed free access to food (controls). The specific binding of 125I labelled bovine growth hormone was low in liver microsomal membranes (45% that of controls) and in plasma membranes (52% that of controls) of fasted rats. The number of somatotropic sites rather than the affinity of the binding was decreased. Lactogenic sites as judged by the binding of 125I-labelled human growth hormone were not significantly reduced in liver membranes of fasted rats. 125I-labelled insulin specific binding was enhanced in microsomal (184% that of controls) and plasma membranes (136% that of controls) of fasted rats; these modifications were associated with a decreased insulinemia. But immunoreactive rat growth hormone levels were not different in plasma of fasted, refed and control animals. Decreased plasma bioassayable somatomedin was associated with the low number of somatotropic binding sites in liver membranes of fasted rats. Somatomedin activity of refed animals was comparable to controls. A significant correlation between the plasma bioassayable somatomedin and the hepatic level of somatotropic binding sites was found. It is proposed that, in fasting, the loss of somatotropic binding sites in the liver is one of the possible causes of the decreased plasma somatomedin bioactivity. PMID- 6295848 TI - Phosphodiesterase induction in Dictyostelium discoideum by inhibition of extracellular phosphodiesterase activity. PMID- 6295849 TI - Mitochondrial DNA copy number in bovine oocytes and somatic cells. PMID- 6295850 TI - Stimulation of protein phosphorylation during fertilization-induced maturation of Urechis caupo oocytes. PMID- 6295851 TI - Membrane potential changes associated with differentiation of enterocytes in the rat intestinal villi in culture. PMID- 6295852 TI - Localization of voltage-sensitive calcium channels along developing neurites: their possible role in regulating neurite elongation. PMID- 6295853 TI - The control of embryonic pigment cell proliferation in culture by cyclic AMP. PMID- 6295854 TI - Evidence against insulin synthesis BY IM-9 lymphocytes. AB - Insulin concentrated from acid/ethanol extracts of IMg lymphocytes by the use of octadecylsilyl (ODS) silica cartridges averaged 0.13 ng/ml cell volume compared with the 0.12 ng insulin/ml cell culture medium. The insulin content of these cells appears to be less than 1% of previously reported values and probably could be accounted for by concentration of insulin from the incubation medium on cell receptors. PMID- 6295855 TI - Restriction fragment length polymorphism of the insulin gene in diabetes mellitus. AB - Variant DNA sequences flanking the human insulin gene were found in the Danish population using restriction endonucleases (restriction fragment length polymorphism). The frequencies of these DNA sequences were determined in 47 non insulin-dependent diabetics and 93 control individuals. We report an association between a restriction fragment length polymorphism of the insulin gene and NIDDM. PMID- 6295856 TI - Impaired insulin secretion in human diabetes mellitus. The effect of naloxone induced opiate receptor blockade. AB - Human diabetes mellitus is characterized by impaired insulin response to intravenous glucose. In search of possible factors which impair insulin release, we have investigated the effect of naloxone, a specific opiate receptor blocker, on insulin responses to glucose in subjects with non-insulin-dependent diabetes, as well as in normal subjects. Naloxone was given as a priming dose of 0.4 mg followed by a constant infusion of either 0.4 mg (N = 7), 2 mg (N = 7), or 4 mg (N = 8) for 90 min. Acute insulin response to glucose (mean change 3-10 min insulin), second phase insulin secretion (change 10-60 min), as well as glucose disappearance rates (%/min) were significantly increased in the diabetics receiving the two higher doses of naloxone (2 and 4 mg, respectively). None of these effects were seen in diabetics receiving saline or in normal subjects receiving naloxone. These results seem to suggest that sensitivity to endogenous opiates may play some part in non-insulin-dependent diabetes. PMID- 6295857 TI - Insulin-induced loss of insulin-like growth factor-I receptors on IM-9 lymphocytes. AB - Preexposure of IM-9 lymphocytes to the somatomedin peptide insulin-like growth factor-I (IGF-I) results in a time- and concentration-dependent reduction in specific receptors for IGF-I. Since insulin and proinsulin are structurally homologous to IGF-I, we investigated the ability of insulin analogues to compete for occupancy and to directly modulate IGF-I receptor concentrations. IGF-I binds rapidly and reversibly to IM-9 cells at 15 degrees C, with half-maximal displacement of 125I-I-IGF-I at IGF-I concentrations of 3.6 X 10(-9) M and insulin concentrations of 5 x 10(-7) M. Preexposure of cells at 37 degrees C to either IGF-I or insulin produced a concentration-dependent reduction in binding of 125I-IGF-I. A 50% decrease in binding was observed following preincubation of cells with IGF-I at 2.5 x 10(-9) M and insulin at 2 x 10(-7) M. At higher insulin concentrations (10(-6)-10(-5) M), up to 70% reduction in 125I-IGF-I binding occurred. Bovine proinsulin and guinea pig insulin competed less potently than porcine insulin for the IGF-I receptor, and produced receptor loss in proportion to their ability to occupy the IGF-I receptor. Scatchard analysis indicated that at all insulin concentrations, the decrease in binding was secondary to loss of available IGF-I receptors, with no change in affinity. Receptor loss was evident following 1-2 h preexposure to insulin, with a t1/2 of 4 h and maximal receptor loss within 10 h. Similarly, IGF-I and IGF-II competed for occupancy of the IM-9 insulin receptor, with 50% displacement of 125I-insulin occurring at peptide concentrations of 3.5 x 10(-9) M (insulin), 3.5 x 10(-8) M (IGF-II), and 3 x 10( 7) M (IGF-I). Preexposure of cells to these peptides at 37 degrees C for 20 h resulted in a concentration-dependent reduction in binding of 125I-insulin, with the order of analogue effectiveness being insulin greater than IGF-II greater than IGF-I. These data emphasize the structural and functional homology of insulin and the somatomedin peptides, IGF-I and II, as well as their respective receptors. Additionally, the data support the conclusion that the insulin and somatomedin peptides not only bind to both receptors, but downregulate each receptor in proportion to their ability to occupy that receptor. PMID- 6295858 TI - Kinetics of somatostatin receptor migration in isolated pancreatic islets. AB - Eighty-seven percent of the total cellular pool of somatostatin (SRIF) receptors in pancreatic islets are located intracellularly. Upon glucose stimulation (300 mg/dl) of insulin release, 8-15% of intracellular SRIF receptors are translocated to the plasma membrane. Affinity of SRIF receptors does not change during their migration and the total cellular pool of receptors remains constant. With prolonged glucose stimulation, surface membrane somatostatin receptor concentration reaches a maximum level at 60 min. PMID- 6295859 TI - Insulin, glucagon, and somatostatin receptors on cultured cells and clones from rat islet cell tumor. AB - Cells grown in culture from rat islet cell tumor (parent cells) and clones obtained from them were used in this study. Parent cells secreted primarily insulin and somatostatin with very small quantities of glucagon. The clones, based on hormone content and secretion, were divided into three phenotypic groups: insulin secreting, somatostatin secreting, and nonsecreting clones. Specific receptors for insulin, glucagon, and somatostatin were demonstrated on parent cells and clones. Parent cells bound 4.12 +/- 0.46% insulin, 2.11 +/- 0.29% glucagon, and 2.49 +/- 1.24% somatostatin per 2 x 10(6) cells. Characteristic hormone binding patterns were observed in insulin secreting versus somatostatin secreting clones. Insulin secreting versus somatostatin secreting clones. Insulin secreting clones bound less insulin than somatostatin and other noninsulin-secreting clones (P less than 0.02). In contrast, somatostatin secreting clones bound more somatostatin than non-somatostatin-secreting clones (P less than 0.05). Somatostatin-secreting clones had a significantly greater number of receptors for all three hormones. The difficulties involved in the interpretation of the quantitative aspects of binding in the presence of continued hormone secretion are discussed. Nonetheless, the presence of receptors on the cells for hormones secreted by the same cells strongly suggests autoregulation. The apparent low affinity of some of these receptors and the presence of receptors for all three islet cell hormones on all islet cells supports the likelihood of paracrine controls. PMID- 6295860 TI - Calcium-calmodulin-dependent myosin phosphorylation by pancreatic islets. AB - Pancreatic islets contain enzyme activity which catalyzes the phosphorylation by MgATP of cardiac, skeletal, or smooth muscle myosin light chains. The enzyme is activated by calcium (Ka = 10 microM) and calmodulin (Ka = 2 nM) and inhibited by trifluoperazine (Ki = 10 microM), a known inhibitor of calmodulin and of insulin secretion. The enzyme binds to a calmodulin affinity column when Ca2+ is present and is eluted when Ca2+ is omitted. These are the properties of myosin light chain kinase. Since phosphorylation of smooth muscle myosin is necessary for its activation by actin, the kinase may have a key role in coupling stimuli that increase intracellular calcium to the contractile processes involved in insulin secretion. PMID- 6295861 TI - Ranitidine in long-term duodenal ulcer treatment: a multicentre trial. PMID- 6295864 TI - Tissue-specific distribution of DNA-binding nuclear proteins. AB - The DNA-binding capacity of nuclear proteins of mouse cells was examined by the protein-blotting method. Under conditions in which the lac repressor specifically binds to the lac operator, the DNA-binding nuclear proteins from different tissues showed a tissue-specific distribution, suggesting that the species and amounts of nuclear proteins with DNA binding activity differ in different tissues. When cloned eukaryotic genes were used for binding, eukaryotic DNA showed stronger binding than prokaryotic DNA. Competition experiments suggested that many nuclear proteins have different DNA binding properties from that of the prokaryotic repressor. PMID- 6295863 TI - Evaluation of steroid biosynthetic lesions in isolated Leydig cells from the testes of streptozotocin-diabetic rats. PMID- 6295865 TI - Reactivity of sera from patients with adult T-cell leukemia (ATL) with type C virus particles associated with ATL cell lines: immunoelectron microscopic study. PMID- 6295862 TI - The diabetic diet, dietary carbohydrate and differences in digestibility. PMID- 6295866 TI - Effects of the tumor promoter teleocidin on human hepatoma cells in tissue culture. AB - The tumor promoter teleocidin inhibited the proliferation of human HUH hepatoma cells in tissue culture. HUH cells cultured with teleocidin developed elongated cytoplasm. In addition, these biological effects of teleocidin were associated with a remarkable decrease in the amount of cellular fibronectin. The synthesis of structural proteins and the number of cell surface receptors for epidermal growth factor were not altered in cells treated with teleocidin, suggesting that teleocidin acted selectively on some of the cellular constituents such as fibronectin. The present data also suggest a possible relationship between the cellular fibronectin content and the biological characters of HUH cells. PMID- 6295867 TI - Receptors for peanut agglutinin isolated from cell lines of human Burkitt lymphoma, lung squamous cell carcinoma and gastric signet ring cell carcinoma. AB - By affinity chromatography on peanut agglutinin-agarose, receptors for peanut agglutinin were isolated from three lines of human malignant cells labeled with [3H]-galactose. The cell lines examined were a Burkitt lymphoma line Daudi, a lung squamous carcinoma QG-56 and a gastric signet ring carcinoma KATO-III. The isolated receptors from the three sources had similar properties and behaved as glycoproteins with apparent molecular weights of 160,000 to 280,000 upon sodium dodecyl sulfate gel electrophoresis. However, they showed molecular weights of only around 70,000 upon gel filtration, suggesting that they contained large amounts of carbohydrates. Pronase digestion depolymerized the glycoproteins, though significant amounts of the products were large enough to be excluded from a column of Sephadex G-50. The large molecular weight products were not detected when the receptors were treated with mild alkali and NaBH4 prior to pronase digestion. Major components of the radioactive oligosaccharides and glycopeptides released in the latter case had molecular weights in the range from 3,500 to less than 1,000. These results indicate that the glycoprotein receptors had clusters of oligosaccharide chains of small and medium sizes. PMID- 6295868 TI - [Male homosexuality and morbidity]. PMID- 6295869 TI - [Association of hepatocellular adenoma and focal nodular hyperplasia of the liver in a woman on oral contraceptives]. PMID- 6295870 TI - Effect of dopamine and bromocriptine on rat ileal and colonic transport. Stimulation of absorption and reversal of cholera toxin-induced secretion. AB - Water and electrolyte transport were determined in rat ileum and colon using the single-pass perfusion technique. Intraperitoneal dopamine caused prompt stimulation of both ileal and colonic water absorption. The dopamine effect was mediated by both specific dopamine and alpha 2-adrenergic receptors. Haloperidol, a specific dopamine antagonist, and yohimbine, an alpha 2-adrenergic antagonist, inhibited the effect of dopamine in ileal absorption; both antagonists alone had no effect on basal water transport. Bromocriptine (intravenous and intraluminal) stimulated ileal and colonic water absorption, which was inhibited by haloperidol and yohimbine, and reversed cholera toxin-induced ileal secretion. Magnitude and time-courses of the increased water absorption in ileal loops, inoculated with saline, were the same as in loops, inoculated with saline, suggesting that bromocriptine acted to reverse cholera toxin-induced secretion by stimulating absorption. Bromocriptine had no effect on the cyclic adenosine monophosphate increase caused by cholera toxin. We conclude (a) dopamine stimulates water absorption in vivo in rat ileum and colon; (b) this dopamine effect is via specific dopamine and alpha 2-receptors; (c) bromocriptine stimulates water absorption in ileum and colon and also acts by dopamine and alpha 2-receptors; and (d) bromocriptine reverses cholera toxin-induced secretion. PMID- 6295871 TI - Carcinoembryonic antigen radioimmunodetection in the evaluation of colorectal cancer and in the detection of occult neoplasms. AB - Radioimmunodetection of colorectal cancer was evaluated in 51 patients by injecting 131I-labeled goat antibody immunoglobulin G against carcinoembryonic antigen and performing total-body photoscans with a gamma scintillation camera 24 and 48 h later. The scintigrams were then processed by computer to subtract the images of the 99mTc-serum albumin and pertechnetate administered, which reflect background and nontarget radioactivity, from the 131I-antibody scans. The results indicate that radioimmunodetection is a safe and a potentially clinically useful cancer detection method, which in this study demonstrated primary colorectal carcinomas in 10 of 12 (83%) of the patients evaluated preoperatively and between 87% (46 of 53) and 92% (49 of 53) of known metastatic tumor sites. Thus, the method's overall sensitivity (true-positive rate) was 86%-91% on a tumor-site basis. A false-negative rate of between 9% and 14% and a false-positive rate of less than 4% were found. In 11 of the 51 patients evaluated, tumor sites were detected that were not found by other clinical methods of cancer detection. These sites of tumor were then confirmed later, as much as 40 wk after radioimmunodetection was performed. It is concluded that in colorectal cancer patients, the current method of carcinoembryonic antigen radioimmunodetection can (a) contribute to the preoperative clinical staging of the patients, (b) assist in the postoperative evaluation of tumor recurrence or spread, (c) complement other methods used to assess tumor response to therapy, (d) support the indication of a rising carcinoembryonic antigen titer (when other methods cannot detect tumor) for second-look surgery, and (e) confirm the findings of other detection measures that are less tumor-specific. PMID- 6295873 TI - Inhibitory characteristics of indomethacin on the activation of K+-dependent p nitrophenylphosphatase by both Na+ and ATP in vitro. AB - 1. The activation of K+-NPPase by both Na+ and ATP was inhibited by indomethacin in vitro. 2. Indomethacin concentration required for 50% inhibition was 6.25 X 10(-4) M. 3. Inhibition manners of indomethacin for Na+ and ATP were competitive type and noncompetitive type, respectively. 4. The Ki values for indomethacin for Na+ and ATP were 1.4 X 10(-4) M and 8.8 X 10(-4) M, respectively. 5. The inhibitory effect of indomethacin was reversible. PMID- 6295872 TI - Interaction of neurotensin, cholecystokinin, and secretin in the stimulation of the exocrine pancreas in the dog. AB - The effects of exogenous neurotensin, secretin, and cholecystokinin-33 alone and in combination on pancreatic secretion were investigated in dogs prepared with pancreatic fistulae. Neurotensin infused intravenously caused a dose-dependent stimulation of exocrine pancreatic secretion. Increasing doses of neurotensin combined with a constant small dose of secretin potentiated pancreatic output of protein and had a tendency to reduce secretion of the bicarbonate. Increasing doses of neurotensin combined with a constant small dose of cholecystokinin-33 potentiated pancreatic output of bicarbonate and lead to a reduction (insignificant) of pancreatic protein secretion. These observations suggest an interaction of neurotensin with pancreatic receptors for secretin and cholecystokinin. PMID- 6295874 TI - Ontogeny of endocrine control of osmoregulation in chick embryo. I. Role of pituitary gland in distribution of water and ions among embryonic and extraembryonic compartments. PMID- 6295875 TI - Ontogeny of endocrine control of osmoregulation in chick embryo. II. Actions of prolactin, arginine vasopressin, and aldosterone. PMID- 6295876 TI - [Cloning the DNA fragments of Streptomyces coelicolor A3(2) plasmid SCP2 in Escherichia coli cells]. PMID- 6295877 TI - [Isolation of mutants with an increased frequency of transposon Tn1 translocations in Escherichia coli K-12 cells]. AB - Two het mutations (high efficiency of transposition) were isolated and characterized preliminarily. The mutations lead to increasing the frequency of ampicillin transposon Tn1 translocation into different replicons in Escherichia coli cells. Both mutations increase the frequency of transposition by the same degree. One of het mutations has been localized within Tn1 transposition element. This mutation seems to be similar to mutations isolated earlier (Gill et al., 1978) in the transposon Tn3 repressor gene which occurred at a high frequency and led to an increase in transposition. The other het mutation is situated in the bacterial chromosome. PMID- 6295878 TI - Transposon Tn3-mediated replicon fusion. AB - To investigate inter-replicon transposition of Tn3, we used the cosmid-phage lambda packaging system coupled with density gradient fractionation and isolated recombinant molecules of different sizes. Cosmids derived from ampicillin resistance-transducing phage were classified into four groups: (1) cosmid-Tn3 donor cointegrates considered as Tn3 transposition intermediates, (2) similar cointegrates carrying deletions of one copy of Tn3 and of adjacent cosmid DNA sequences, (3) cosmids carrying a single Tn3 insertion, and (4) cosmids carrying two independent Tn3 insertions. Genetic and biochemical studies indicated that cosmids isolated from ampicillin-resistance transductants were derived from the authentic cosmid-Tn3 donor cointegrate intermediates. PMID- 6295879 TI - The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. AB - A series of plasmid vectors containing the multiple cloning site (MCS7) of M13mp7 has been constructed. In one of these vectors a kanamycin-resistance marker has been inserted into the center of the symmetrical MCS7 to yield a restriction-site mobilizing element (RSM). The drug-resistance marker can be cleaved out of this vector with any of the restriction enzymes that recognize a site of the flanking sequences of the RSM to generate an RSM with either various sticky ends or blunt ends. These fragments can be used for insertion mutagenesis of any target molecule with compatible restriction sites. Insertion mutants are selected by their resistance to kanamycin. When the drug-resistance marker is removed with PstI, a small in-frame insertion can be generated. In addition, two new MCSs having single restriction sites have been formed by altering the symmetrical structure of MCS7. The resulting plasmids pUC8 and pUC9 allow one to clone doubly digested restriction fragments separately with both orientations in respect to the lac promoter. The terminal sequences of any DNA cloned in these plasmids can be characterized using the universal M13 primers. PMID- 6295880 TI - A new pair of M13 vectors for selecting either DNA strand of double-digest restriction fragments. AB - The strategy of shotgun cloning with M13 is based on obtaining random fragments used for the rapid accumulation of sequence data. A strategy, however, is sometimes needed for obtaining subcloned sequences preferentially out of a mixture of fragments. Shotgun sequencing experiments have shown that not all DNA fragments are obtained with the same frequency and that the redundant information increases during the last third of a sequencing project. In addition, experiments have shown that particular fragments are obtained more frequently in one orientation, allowing the use of only one of the two DNA strands as a template for M13 shotgun sequencing. Two new M13 vectors, M13mp8 and M13mp9, have been constructed that permit the cloning of the same restriction fragment in both possible orientations. Consequently, each of the two strands becomes a (+) strand in a pair of vectors. The fragments to be cloned are cleaved with two restriction enzymes to produce a fragment with two different ends. The insertion of such a fragment into the vector can occur only in one orientation. Since M13mp8 and M13mp9 have their array of cloning sites in an antiparallel order, either orientation for inserting a double-digest fragment can be selected by the choice of the vector. PMID- 6295881 TI - Insertional mutagenesis in Bacillus subtilis: mechanism and use in gene cloning. AB - The plasmid pHV32, which replicates in Escherichia coli but not in Bacillus subtilis, transformed B. subtilis-competent cells efficiently when linked in vitro to EcoRI B. subtilis DNA segments. The transformed clones carried pHV32 inserted in their chromosomes, and often displayed a mutant phenotype. One of the transformed clones carried pHV32 inserted close to the thyB gene. We cleaved the DNA extracted from this clone with BglII restriction endonuclease, for which no sites exist on pHV32, ligated the released segments and used them to transform E. coli selecting for pHV32-carried genetic markers. The transformants harbored a hybrid plasmid which carried the B. subtilis thyB gene. Circular molecules composed of pHV32 joined to B. subtilis DNA inserted into the chromosome by a Campbell-like recombination event. Linear molecules, in which pHV32 was flanked by two non-adjacent DNA segments, underwent a double cross-over recombination with the chromosome. In this case the chromosomal sequences between the non adjacent segments were deleted, and replaced by pHV32 sequences. PMID- 6295882 TI - Probing cII and himA action at the integrase promoter pi of bacteriophage lambda. AB - Plasmids were constructed to supply cII-coded protein for activation of the phage promoter pI. Using a fusion which expresses lacZ from pI. We can accurately follow activation of pI without having to assay int activity in vivo. A large excess of cII protein compared to a normal lytic infection stimulates lacZ expression about 10-fold over the basal level. The int-c226 constitutive allele of pI is not further activated by cII even though its level of lacZ expression is less than the maximal cII-activated expression from wild type pI. A himA-deleted strain also shows activation, demonstrating that there is no absolute requirement for the himA gene product for cII-stimulated transcription at pI. PMID- 6295883 TI - Cloning of alkaline phosphatase isozyme gene (iap) of Escherichia coli. AB - In Escherichia coli three major alkaline phosphatase isozymes are formed by molecular conversions depending on physiological conditions. A chromosomal gene, iap, is responsible for alkaline phosphatase isozyme conversion and is assumed to code for a proteolytic enzyme removing the arginine residue(s) from the N terminal position of alkaline phosphatase subunits. A chromosomal fragment which complemented the Iap- phenotype was cloned into pBR322 by a shotgun method. Transducing phage lambda iap was constructed in vitro from the chromosomal fragment containing the iap gene and lambda tna DNA. The integration site of the phage on chromosome was identified as the iap locus by P1 transduction, which meant that the cloned chromosomal DNA contained authentic iap gene. The restriction map of the hybrid plasmid was constructed. Based upon this information, several iap deletion plasmids as well as smaller iap+ plasmids were constructed. Analysis of the phenotypes conferred by these plasmids enabled us to locate iap gene within a 2-kb segment of the cloned DNA. The cells carrying the iap+ plasmid showed very efficient isozyme conversion even in medium containing arginine, an inhibitor for the isozyme conversion. This indicates overproduction of the iap gene product. PMID- 6295884 TI - Nucleotide sequence and exact localization of the neomycin phosphotransferase gene from transposon Tn5. AB - The nucleotide sequence of 1200 bp from the unique region of transposon Tn5 containing the neomycin phosphotransferase gene (neo) was determined, and the location of the neo gene was identified by deletion mutants in a translational reading frame of 792 bp. The derived gene product, an aminoglycoside 3' phosphotransferase (APH) II, consists of 264 amino acid residues and has a calculated Mr of 29053. Its amino acid sequence shows sequence homologies to the APH type I enzyme coded for by transposon Tn903 (Oka et al., 1981). PMID- 6295885 TI - Expression of cloned calf prochymosin gene sequence in Escherichia coli. AB - An expression plasmid for calf prochymosin (prorennin) cDNA was constructed. The plasmid (pCR301) contains the lacUV5 promoter in front of the fused gene in which the codons for the N-terminal four amino acids of prochymosin cDNA were replaced with those for the N-terminal ten amino acids of beta-galactosidase. Synthesis of the fused protein with the expected Mr was detected immunologically in Escherichia coli harboring pCR301. The product seemed to be localized in the cell membrane of the bacterial host. PMID- 6295886 TI - A cloning vector able to replicate in Escherichia coli and Streptococcus sanguis. AB - A plasmid that is able to replicate in both Escherichia coli and Streptococcus sanguis has been constructed by the in vitro joining of the pACYC184 (Cmr Tcr) and pVA749 (Emr) replicons. This plasmid, designated pVA838, is 9.2 kb in size and expresses Emr in both E. coli and S. sanguis. Its Cmr marker is expressed only in E. coli and may be inactivated by addition of DNA inserts at its internal EcoRI or PvuII sites. The pVA838 molecule also contains unique SalI, SphI, BamHI, NruI and XbaI cleavage sites suitable for molecular cloning. pVA838 may be amplified in E. coli but not in S. sanguis. We have used the pVA838 plasmid as a shuttle vector to clone streptococcal plasmid fragments in E. coli. Such chimeras isolated from E. coli were readily introduced into S. sanguis by transformation. PMID- 6295887 TI - Cloning of Pseudomonas plasmid pMG7 and its restriction-modification system in Escherichia coli. AB - Plasmid pMG7 of Pseudomonas aeruginosa codes for a type II DNA restriction modification (r-m) system, PaeR7. This plasmid has not been observed to transfer to Escherichia coli either by conjugation or by transformation. We have cloned BglII linears (42 kb) and the BamHI large fragment (37 kb) of pMG7 into cosmid pHC79 (6.4 kb) and introduced the recombinant molecules into E. coli by in vitro packaging. Several clones were obtained which demonstrated in vivo restriction of phage phi 80. One of these clones, GT138, was further tested and showed in vivo modification of phi 80. Extracts from two clones, GT138 and GT125, yielded a restriction endonuclease activity which produced fragments of phi 80 DNA identical to those produced by PaeR7. Cosmid cloning should be useful for obtaining substantial yields of large fragments of plasmids that are difficult to purify in their native strains. PMID- 6295888 TI - Broad-host-range cloning vectors derived from the W-plasmid Sa. PMID- 6295889 TI - Selection and follow-up of patients for conservation surgery and irradiation. PMID- 6295890 TI - Immunoperoxidase estrogen receptor assay for human breast cancer. PMID- 6295891 TI - [Hygienic bases for using the wastes from hydroxyacetyl compound production as fertilizer]. PMID- 6295892 TI - [Morphofunctional changes in the rat peripheral nervous system during separate and combined exposure to carbon disulfide and noise]. PMID- 6295893 TI - [Blood picture in pneumoconiosis in relation to the silicon dioxide content of the dust]. PMID- 6295894 TI - Hormone regulation of the growth of endometrial hyperplasias and tumors from the aged fischer rat. PMID- 6295895 TI - The influence of topical vaginal estrogen application on the lipids metabolism. PMID- 6295896 TI - A pharmacological approach to the inhibition of platelet adhesion and platelet aggregation. Wright-Schulte Lecture. AB - A first level of pharmacological interference with platelet function is located at the site of the agonist-receptor interaction (receptors for collagen, adenosine diphosphate, serotonin, fibrinogen). A second level of interaction is to prevent the mobilization of intracellular calcium ions which can be obtained by inhibition of phosphodiesterase, activation of cAMP (e.g. with prostacyclin analogues), inhibition of thromboxane A2 formation or blocking of its receptors. Compounds preventing the platelet contractile process or platelet secretion could theoretically also prevent some platelet functions. PMID- 6295897 TI - [Effects of 2,3-dimethoxy-5-methyl-6-(10'-hydroxydecyl)-1,4-benzoquinone (CV 2619) on myocardial energy metabolism in the hypertrophied heart of spontaneously hypertensive rats]. AB - Effects of CV-2619 (10 and 30 mg/kg/day, p.o.) or ubiquinone-10 (Q-10, 10 mg/kg/day, p.o.) treatment for 5 weeks on systolic blood pressure (SBP) and myocardial energy metabolism were studied in spontaneously hypertensive rats of 20 weeks of age. The systolic blood pressure was about 205 mmHg at the start of the experiment, and a slight increase was noted thereafter in the control (vehicle) group. CV-2619, but not Q-10, inhibited the increase in the blood pressure. At 25 weeks of age, cardiac hypertrophy was noted to the same extent in either treated group. Myocardial contents of glycolytic intermediates (glycogen, glucose, pyruvate and lactate) and creatine phosphate (Cr-P), ATP, ADP, and AMP were not significantly influenced by CV-2619 or Q-10 treatment. CV-2619, however, significantly increased the energy charge, an index of myocardial energy state, with higher dose and lowered the lactate/pyruvate ratio with either dose. These results suggest that CV-2619 has a mild antihypertensive effect and improves the myocardial energy state in the hypertrophied heart during the sustained phase of hypertension in SHR rats. PMID- 6295898 TI - [133Xe-labeling of tissue adhesives for catheter embolization]. AB - A method for radio-nuclide labelling of histo-acryl/lipiodol with 133Xe gas is described. The method consists of dissolving the gas in lipiodol under sterile conditions. After embolization and angiography, the localisation of the embolizing material can be determined by means of a scintillation camera. The labelling method was used in 15 patients. In all patients the position of the embolizing material, as demonstrated by scintigraphy, corresponded with the radiological findings. It was not possible to demonstrate escape of the material by scintigraphy. PMID- 6295899 TI - [Computer tomographic densitometry of primary liver tumors]. AB - The dynamic computed tomography (CT) densitometry is defined as the combination of rapid bolus injection of urographic contrast medium and timed sequential computed tomography scan permitting recognition of different patterns of enhancement. The results of this method on fifty patients with histologically proofed hepatic tumors (focal nodular hyperplasia n = 12, cavernous hemangioma n = 13, liver cell carcinoma in cirrhotic liver n = 10, primary liver cell carcinoma n = 6, bile duct carcinoma n = 4, others n = 5) are demonstrated. CT densitometry offers valuable aid for the differential diagnosis of focal liver lesions, i.e. cavernous hemangioma, focal nodular hyperplasia of the liver. PMID- 6295900 TI - [Affection of the nervous system in Behcet's disease--the clinical and neuropathological picture]. PMID- 6295901 TI - Relation of serum alkaline phosphatase to liver scintigram in patients with hepatocellular carcinoma. AB - Serum Alkaline Phosphatase (ALP) was studied in relation to liver scintigrams of 54 patients with hepatocellular carcinoma. The ALP activity was higher with larger tumors and in multiple tumors. Within the single tumor group, the activity was higher when the tumor was located in the hilum than in the periphery. The incidence of ALP-1 isoenzyme (bile ALP) roughly paralleled the total ALP activity. These results suggest that the variation of serum ALP seen in each individual patients with hepatocellular carcinoma reflects the volume of cholestatic liver tissue, which is changed by the number, size and localization of the tumor nodules in the liver. PMID- 6295902 TI - Gastric cytoprotection by antacids and papaverine in rats. AB - Prostaglandin E2 (PGE2) prevented hemorrhagic ulceration of rat stomach mucosa induced by various procedures when given orally at a non-antisecretory dose. This effect of PGE2 is called gastric cytoprotection. We used absolute ethanol, 0.6 N hydrochloric acid and 0.2 N sodium hydroxide as damaging agents. Ranitidine at an antisecretory dose did not exhibit any cytoprotective effect. However, the poorly absorbable antacids, magnesium hydroxide plus aluminium hydroxide and aluminium phosphate inhibited the development of hemorrhagic lesions significantly. A similar protective effect was seen after intragastric administration of papaverine, which is known to stimulate endogenous prostaglandin synthesis. However, the question as to whether or not stimulation of endogenous prostaglandin synthesis is the mode of action of the cytoprotective effect of papaverine and poorly absorbable antacids, cannot yet be answered. PMID- 6295903 TI - Preincubation and purification abolish the hypophysectomy induced hypersensitivity of isolated rat adrenal cells. PMID- 6295904 TI - Some factors affecting glomerulopressin production by perfused rat liver. AB - Studies were carried out on the production of the glucuronide hormone glomerulopressin by the isolated rat liver in male and female rats, and the effect on this production of an inhibitor (Novobiocin) and a stimulator (benzo (a)pyrene) of glucuronyltransferase activity. It was observed that males produce more glomerulopressin than females, Novobiocin inhibits its production while benzo-(a)pyrene enhances it in females but not in males. The changes in glomerulopressin production are related to the changes in glucuronyltransferase activity observed by other authors, suggesting that this enzyme is involved in the production of glomerulopressin. PMID- 6295905 TI - Characterization of [125I]-iodo-ovine prolactin and evaluation of prolactin receptor assay methods. AB - In an attempt to establish more rigorous conditions for assay of prolactin receptor, a study of the preparation, recovery and specific binding of iodoprolactin was conducted. Averaging the results of 20 preparations starting with 5 micrograms prolactin preparation and iodinated by a lactoperoxidase method, an average of 0.62 +/- 0.12 micrograms of the iodinated prolactin was recovered in a form capable of binding to excess rat liver membrane preparations, corresponding to between 24 and 51% of the total recovered iodinated prolactin. Based on preliminary amino acid analysis and N-terminus determination, 53% of a weighed amount of ovine prolactin was determined to be protein with the N terminal amino acid being consistent with homogeneity. The percent specific binding assay (single point assay) and Scatchard analysis were tested for their ability to give assay results in a direct relationship to sample receptor. Scatchard analyses were determined to be a superior method to the percent specific binding assay. Using commonly employed conditions for the percent specific binding assay, the relative amounts of receptor found in increasing amounts of sample were significantly less than that found by Scatchard analysis in all likelihood due to the use of subsaturating concentrations of iodohormone. As more hormone was added per assay tube, the relationship of sample receptor to the results of the single point assay improved, but the results never equaled those of Scatchard analyses. Thus, many workers who have used the percent specific binding assay under nonsaturating conditions have underestimated the relative receptor contents of samples. PMID- 6295906 TI - Modulation by S-adenosyl-L-methionine of hepatic Na+,K+-ATPase, membrane fluidity, and bile flow in rats with ethinyl estradiol-induced cholestasis. AB - Structural and functional changes in the surface membranes of hepatocytes play a pivotal role in the induction and reversion of some forms of drug-induced cholestasis. To elucidate the mechanism by which S-adenosyl-L-methionine (SAMe) leads to a partial reversion of bile flow impairment caused by ethinyl estradiol (EE), female Sprague-Dawley rats were given oral doses of EE (5 mg per kg per day, for 3 days) with and without simultaneous administration of SAMe (25 mg per kg, 3 times per day, for 3 days). Na+,K+-ATPase activity and membrane microviscosity as measured by fluorescent polarization were assayed in isolated liver plasma membranes (LPMs). SAMe administration to normal and EE-treated rats resulted in a marked increase in Na+,K+-ATPase activity and LPM fluidity. EE alone did not cause any change in the physicochemical properties of the LPMs. Hepatic Mg2+-ATPase and gamma-glutamyl transpeptidase activities were not affected by SAMe alone but increased when SAMe was given together with EE. These data indicate that the interaction of in vivo administered SAMe with hepatocyte plasmalemma and its effect on lipid fluidity and enzymes of the LPMs showed a high specificity and an inverse relationship between Na+,K+-ATPase activity and fluorescence polarization values. Furthermore, modulation of hepatic Na+,K+ ATPase was associated with SAMe-induced protection against bile flow impairment due to EE; however, it was not the causative factor for EE-induced cholestasis under the experimental conditions. These findings suggest that changes in surface membrane structure and function might account in part for the reversal by SAMe of EE-induced impairment of bile secretory function. PMID- 6295907 TI - Effect of chronic ethanol feeding on hepatic collagen in the monkey. AB - The effect of chronic ethanol feeding was determined on parameters of hepatic collagen metabolism in the monkey. Four monkeys of the species Macaca radiata received a nutritionally adequate diet containing 50% of the calories as ethanol, while four others were pair-fed a diet in which ethanol was isocalorically substituted by carbohydrate. Liver biopsies were obtained at 3, 12, and 24 months, and the animals were killed between 40 and 48 months after initiation of the diets. The ethanol-fed animals developed various degrees of fatty infiltration, but no necrosis, inflammation, or fibrosis. The amount and distribution of collagen Types I, III, and IV demonstrated by immunohistochemical techniques was not altered after ethanol feeding. No changes were found in hepatic protein-bound hydroxyproline or in collagen prolyl hydroxylase activity at the various time intervals. Liver free proline and the incorporation of labeled proline into protein-bound hydroxyproline by liver slices were not altered after 40 to 48 months of ethanol feeding. This study shows that prolonged feeding of ethanol together with an adequate diet results in fatty infiltration but not deposition or alteration of hepatic collagen metabolism. PMID- 6295908 TI - Hepatitis B virus status of southern African Blacks with hepatocellular carcinoma: comparison between rural and urban patients. AB - Hepatocellular carcinoma (HCC) is less common and occurs at a much older age in urban than in rural southern African Blacks. These differences may reflect differences in the etiology of the tumor in the two populations. The purpose of this study was to compare the hepatitis B virus (HBV) status of 150 HCC patients who were born and had lived all their lives in a rural environment with 158 patients who were born and brought up in a rural setting but then became urbanized. HBsAg and all markers of present or past HBV infection [HBsAg(+) or anti-HBc(+) or anti-HBs] were significantly less common in the urban patients when the two groups were considered as a whole (p less than 0.001 and p less than 0.05, respectively). However, because the rural patients were considerably younger (mean age 34.7 years; 66% less than 40 years of age) than in urban patients [mean age 50.9 years (p less than 0.0005), 19.0% less than 40 years of age (p less than 0.001)], an age-related analysis was performed. No significant difference in any HBV marker was found between rural and urban patients. The association between active HBV infection and HCC was similar in young patients, both rural and urban, and the prevalence of HBs antigenemia decreased in both groups with increasing age. We conclude that the differences in incidence and age of onset of HCC in rural and urban southern African Blacks cannot be attributable to differences in HBV status. PMID- 6295909 TI - Relationship of pretransplant hepatobiliary disease to bile duct damage occurring in the liver allograft. AB - Histopathological features of nonsuppurative destructive cholangitis have been described in primary biliary cirrhosis, chronic graft-vs-host disease, and chronic rejection of human liver allografts. To determine whether or not susceptibility to injury of interlobular bile ducts was related to the original hepatobiliary disease requiring transplantation, we compared the histopathology of allografts transplanted into two groups of patients. The first group consisted of patients whose original hepatobiliary diseases primarily affect bile ducts: primary biliary cirrhosis, sclerosing cholangitis, and biliary atresia. The second group consisted of patients whose original hepatobiliary diseases do not result in injury to interlobular bile ducts. In 32 liver allografts studied histopathologically, the original disease did not recur. Interlobular bile duct damage occurred, however, in 17 of the 32 and resembled that seen in primary biliary cirrhosis. The spectrum of bile duct injury included mononuclear inflammation of the bile duct epithelium, destruction of the bile duct epithelium, and dissolution of the bile duct. There was no relationship between the frequency or severity of bile duct damage and original hepatobiliary disease, age, sex, duration of allograft survival, or cause of death. We conclude that nonsuppurative destructive cholangitis occurs frequently in liver allografts and represents one component of allograft rejection. This lesion resembles, in many respects, that seen in primary biliary cirrhosis so that histopathological differentiation may be difficult. PMID- 6295911 TI - A mutual-performance contract to close the gap between hospital and community. PMID- 6295910 TI - Dieldrin-induced mallory bodies in hepatic tumors of mice of different strains. AB - Mallory bodies (MBs) were induced in hepatic tumors by administration for up to 85 weeks of a diet containing 10 ppm dieldrin to 50 C3H/He and 62 C57BL/6J x C3H/He B6C3F1 male mice. MBs were seen in 15 of 28 (54%) mice which developed benign hepatic tumors and 33 of 45 (73%) mice with hepatocellular carcinoma, but in only 3 of 39 (8%) mice without hepatic tumors. In mice with tumors, the MBs were predominantly confined to tumor tissue and persisted in a carcinoma transplanted into a nude mouse. MBs were not observed, however, in hepatic tumors of 67 C57BL/6J, 49 C3H/He, or 81 B6C3F1 mice given 12 micrograms diethylnitrosamine i.p. on Days 0, 3, 9, and 15. Thirty-one of 195 control mice of all three strains had hepatic tumors. Only one of the controls had a tumor with an MB, and no MBs were seen in nontumor-bearing livers of controls animals. These observations, coupled with the results of a previous study in which MBs were observed in hepatocytes of dieldrin-treated C57BL/6J mice, indicate that mice treated with dieldrin are a reliable animal model for the study of MBs. PMID- 6295912 TI - Oregon's innovative system for supervising offenders found not guilty by reason of insanity. PMID- 6295913 TI - Enders, Weller, and Robbins: the trio that 'fished in troubled waters'. PMID- 6295914 TI - Sexually transmitted diseases. AB - The "traditional" venereal diseases account for a small fraction of the sexually transmitted diseases prevalent in industrialized societies. The clinical as well as the public health impact of the widening range of STDs is discussed. PMID- 6295915 TI - Xanthomatous neuropathy of liver. AB - A 36-year-old man with juvenile-onset diabetes mellitus developed cholestatic hepatitis of unknown cause, possibly drug-induced. He remained jaundiced for four months, until he died unexpectedly. At autopsy, severe and macroscopically identifiable xanthomatous neuropathy was found in the liver; it involved the unmyelinated nerves in the hilus and in all portal tracts examined. Microscopic evidence of minimal extrahepatic involvement of unmyelinated nerves was also found. Somatic nerves were affected by diabetic neuropathy but not by xanthomatous neuropathy. The condition seems to be a rare complication of diseases that are characterized by accumulations of lipids. PMID- 6295916 TI - Immunohistochemistry of soft tissue tumors: progress and prospects. AB - The immunohistochemical definition of soft tissue tumors has only begun (table 1). A number of antigens described here play a role in diagnostic pathology, and a few have been shown to be specific markers for certain types of cell differentiation. The detection of a wide variety of antigens in soft tissue sarcomas may initially prove confusing in some areas. For example, when specific markers are used, multidirectional differentiation in a tumor may be found more frequently than it is now, making more sarcomas actually "mesenchymomas." In such situations, certain types of differentiation may imply more or less aggressive behavior. It is also possible that panels of different immunohistochemical reagents may be required to determine a specific histogenesis. Furthermore, interpretive caution is clearly necessary, since many sarcomas contain normal tissues. Owing to the aggressive nature of sarcomas, more information on their biochemical composition will be required for proper diagnosis and clinical management. As pathologists, we are in a key position to contribute to the understanding of these tumors. In-situ biochemical analysis of sarcomas making use of immunohistochemical methodology is a powerful tool in the investigation of the difficult problems of histogenesis, tumor heterogeneity, and biologic potential. PMID- 6295917 TI - Intracellular distribution of DNA topoisomerase I in fibroblasts from patients with Fanconi's anaemia. AB - The activity of DNA topoisomerase I(DNA nicking-closing enzyme) was analysed in cytoplasmic and nuclear extracts of six independently derived Fanconi and four normal fibroblast cell lines. In all experiments the total cellular activity was predominantly found in the nuclear extracts (88-100%). In addition, a minor proportion of the enzyme (up to 12%) was randomly present in some of the cytoplasmic fractions of both Fanconi and normal fibroblasts. These results indicate that Fanconi's anaemia is probably not due to or accompanied by a maldistribution of topoisomerase I between nuclei and cytoplasm. PMID- 6295918 TI - Human red cell adenylate kinase polymorphism in Srbija, Yugoslavia. AB - Red cell adenylate kinase (AK) phenotypes were determined in 283 unrelated adults in Srbija (Yugoslavia). The gene frequencies observed were: AK1 0.961 and AK2 0.039. The adenylate kinase activity was estimated in all haemolysates; no significant differences were found between individuals of different phenotypes. PMID- 6295919 TI - The effects of 3',5' adenosine monophosphate on the proliferation of Reuber H35 rat hepatoma cells in vitro. AB - 3',5' Adenosine monophosphate (cAMP) inhibits the proliferation of Reuber H35 rat hepatoma cells at concentrations higher than 10(-5) M. This inhibitory effect can be demonstrated both in exponentially growing monolayer cultures and in single cell clonal growth. The inhibition of the cell proliferation is due to both a short term (division delay) and a long term effect (cytotoxicity). The short term effect seems to be due to cAMP itself as it is potentiated by the phosphodiesterase inhibitor 1-methyl,3-isobutyl xanthine (MIX). The long term effect probably is due to degradation products of the cyclic nucleotide. It is shown by a combination of time lapse cinematography, autoradiography, and flow cytofluorometry that the division delay is due to a prolongation of the S phase with no apparent changes in the duration of the G1 phase. The possible causes of this prolongation of the S phase by cAMP are discussed. PMID- 6295920 TI - Characterization of isolated Fe-loaded rat hepatocytes prepared by collagenase perfusion. AB - Hepatocytes from livers of rats loaded by Fe-dextran treatment were isolated by an in situ collagenase perfusion technique and evaluated for their biochemical, cytochemical, and morphological characteristics in cell culture. Iron loads 15 times higher than in normal rat liver cells isolated in the same way were retained in the preparations with 40% present as hemosiderin. A simple centrifugation-mathematical approach is described for the calculation of Fe content in the hepatocyte (95%) and reticuloendothelial (5%) fractions in the isolates. The cells were cultured for 22 h without loss of protein synthesis capability or significant changes in cell count, viability, endogenous glutamate oxaloacetate transaminase (GOT) or Fe and were morphologically similar in most respects to unloaded (normal) hepatocytes similarly cultured. Studies are in progress to assess the utility of these preparations as a model for Fe mobilization from Fe-loaded animals. PMID- 6295921 TI - Ascorbic acid accumulation by cultured rat adrenocortical cells. AB - The influence of adrenocorticotrophin (ACTH) on radiolabeled ascorbic acid (AA) accumulation by adrenocortical cells was examined in primary cultures of collagenase dissociated glands from adult male rats. The cells were ACTH responsive by morphological and steroidogenic criteria. After 5 d in AA-free medium, cells pretreated with 100 mU/ml ACTH for 3 d took up two to three times more AA over a 2 h period than did untreated controls (4.0 to 10.0 nmol versus 1.7 to 3.4 nmol AA/micrograms DNA). In contrast, ACTH administered on Day 6 concurrently with AA inhibited AA accumulation compared to cultures exposed to AA alone. This acute inhibitory effect of ACTH was in the order of 30% in cultures pretreated with ACTH for 3 d but was not significant (7%) without ACTH pretreatment. The results show that ACTH has distinct long term stimulatory and acute inhibitory effects on AA accumulation by adrenocortical cells and suggest that both maximal AA accumulation and the responsiveness to acute inhibition of AA accumulation by ACTH may depend on the maintenance of the differentiated state of the adrenal cortex. PMID- 6295923 TI - Differential ultrastructure and electron histochemistry of human leukaemic cells. I. Acute and chronic myeloid leukaemias. PMID- 6295922 TI - Chemical carcinogen-mouse mammary tumor virus interactions in cell transformation. AB - We have studied the process of mammary cell transformation in vitro using a single cell clone (Clone 18) from a presumptive epithelial cell line, C57MG, derived from a normal mammary gland; a mouse mammary tumor virus (MMTV) host range variant (RIII)vp4; and the potent initiating carcinogen 7,12 dimethylbenz(a)anthracene (DMBA). After several serial subcultures, cells treated with virus and then with carcinogen exhibited an altered (transformed) morphology, a dramatic increase in anchorage independence, an increase in multinucleation after exposure to cytochalasin B, an enhanced ability to proliferate in low Ca2+ (0.01 mM) medium, and tumorigenicity when inoculated subcutaneously into athymic (nude) mice. Although some of these phenotypic alterations were observed also in cultures treated singly with MMTV or DMBA and in cultures exposed to DMBA before infection with MMTV, enhanced cytochalasin B multinucleation and tumorigenicity were properties observed only in mass cultures of cloned cells first infected with MMTV and then exposed to DMBA. This demonstrates for the first time that exposure of presumptive mammary epithelial cells to MMTV followed by DMBA, but not to either agent alone or to DMBA followed by MMTV, results in malignant transformation of these cells. PMID- 6295924 TI - Retrospective analysis of 31 cases of Wilm's tumour. PMID- 6295925 TI - Melanotic neuroectodermal tumour of infancy. PMID- 6295926 TI - Mechanisms of tumour cell destruction by PMA-activated human neutrophils. AB - Phorbol myristate acetate (PMA)-activated neutrophils were found to destroy B lymphoblast tumour cells (Raji) as determined by the 51Cr release assay. The target cell lysis was prevented by azide, suggesting the involvement of the myeloperoxidase enzyme. Catalase and cytochrome c caused a marked impairment of the neutrophil-mediated cytolysis, whereas superoxide dismutase significantly enhanced the target cell destruction. These data indicate that hydrogen peroxide plays a key role in the target cell injury; superoxide anion appears to be devoid of direct cytotoxic activity, despite its requirement as a precursor of hydrogen peroxide. The target cell destruction required the presence of the iodide ion as oxidizable co-factor for the myeloperoxidase-hydrogen peroxide system. The chloride ion alone was uneffective. Inhibition of target cell metabolic pathways, involved in the cellular defences against oxidative injury, by the anti neoplastic agent 1,3-bis-(2-chloroethyl)-1-nitrosurea (BCNU) resulted in an increased neutrophil-mediated cytolysis. Under the experimental conditions employed, PMA-activated neutrophils incubated with BCNU-treated Raji cells became cytotoxic also in the presence of the chloride ion alone as myeloperoxidase co factor. Our results suggest that Raji target cell destruction by PMA-activated neutrophils depends on the myeloperoxidase-hydrogen peroxide-halide system. The cytolytic event is influenced by target cells themselves, which should be regarded as an active component of the cytotoxic system, capable of interfering with the lytic mediators of the effector cells. PMID- 6295927 TI - Spin labeling of immunoglobulin M and E carbohydrates. AB - The analysis of ESR spectra of spin-labeled human myeloma immunoglobulins M and E has shown the rotation of spin labels bound to carbohydrates to be restricted most probably due to close attachment of oligosaccharide chains to protein parts of molecules. The values of rotational correlation times for IgM, IgMs and Fc5, spin-labeled at carbohydrates, were found to be equal to 7,7 and 6 ns, respectively. These data pointed to the existence of internal lability of the Fc5 fragment. ESR spectra of IgE spin-labeled at carbohydrates also reflected restricted rotation of the spin label, but the presence of a more than one wide extremum spoke in favour of differences in the degree of immobilization between different oligosaccharide chains. The value of rotational correlation times calculated for IgE, spin-labeled at protein and carbohydrate moieties, were similar (60 and 66 ns). These data confirmed the previously found rigidity of IgE molecule as compared with IgG molecule. PMID- 6295928 TI - Activity of Japanese encephalitis virus among certain domestic animals in West Bengal. PMID- 6295929 TI - Viral antibody titres in non-systemic glomerular disease. PMID- 6295930 TI - Virulence, haemolysis pattern and haemophagocytosis index of Entamoeba histolytica. PMID- 6295931 TI - Clinical genetics of human cancer. PMID- 6295932 TI - Environmental modifiers in carcinogenesis. AB - The natural history of neoplastic development can be divided into at least three distinct stages - initiation, promotion and progression. These stages can be defined by biological experimentation and by histopathological characterization of the lesions involved in the various stages. It is now possible, both in human beings and in experimental animals, to distinguish by morphological techniques all stages from very early 'precursor' lesions to the final malignant state. Although the final stage - progression - of the neoplastic process can be altered by therapeutic intervention, the principal site at which the natural history of neoplastic development can be actually and potentially regulated is that of promotion. Promoting agents that are relatively selective for carcinogenesis in several different tissue types have now been identified both in experimental systems and in human beings. In at least two experimental systems - those of epidermal carcinogenesis and hepatocarcinogenesis - the general characteristics of both initiation and promotion are virtually identical. Such studies have demonstrated that the activity of promoting agents exhibits a threshold level as well as a maximal level and that the action of promoting agents is reversible; the identification of promoting agents in our environment is therefore of considerable importance in understanding the nature of and preventing the development of human cancer. In addition, the interpretation of whole-animal bioassays for carcinogenic agents in the environment must take into account the existence of promoting agents and recognize that their action is distinct from that of complete and incomplete carcinogens, in order rationally to extrapolate to situations of human risk. PMID- 6295933 TI - Hormones and the single cell: a relationship prerequisite for transformation. AB - The direct oncogenic potential of radiation and chemicals is demonstrable in cell cultures, where host-mediated influences do not prevail. These systems afford the opportunity of investigating factors that modulate neoplastic transformation. Agents such as retinoids modulate late events and inhibit the expression of transformation and the promotional effects of 12-O-tetradecanoylphorbol-13 acetate. Thyroid hormones play a role in early events, in initiation of transformation. Interaction of triiodothyronine (T3) in a culture medium with single cells is a prerequisite for the initiation of transformation following exposure to X-rays and to benzo[a]pyrene. When cloned hamster embryo cells and mouse 10T 1/2 cells are maintained in a medium containing serum without thyroid hormones (hypothyroid conditions) and exposed to 3 or 4 Gy X-rays, no transformation is observed, although cell survival and cell growth are unaffected by thyroid hormone level. Supplementation of the medium with 10-12M to 10-7M T3 12 h before exposure to X-rays results in a transformation frequency related to the dose of T3, with a peak at 10-10M. The curve is similar to that induced by Na/K ATPase. Addition of T3 at the time of radiation results in a lower transformation frequency; if it is added after radiation no transformation is observed. The effect of T3 in the process of initiation is not mimicked by reverse T3 and is abolished by addition of 100 ng cyclohexamide. The data suggest that the effect of T3 in rendering the cell competent to transform involves synthesis of a cellular 'transforming protein'. Ongoing experiments indicate that exposure to 1 microgram/ml benzo[a]-pyrene results in transformation only in the presence of T3. Under hypothyroid conditions, no transformation is observed. PMID- 6295934 TI - Does hepatitis B virus cause hepatocellular carcinoma? AB - Evaluation of the hypothesis of an association between hepatitis and hepatocellular carcinoma (HCC), first suspected on pathologic grounds, was made easier by the discovery of hepatitis B surface antigen. Population correlation and analytical epidemiologic studies established that there is a strong and specific association between hepatitis B virus (HBV) and HCC. The association is restricted to chronically active forms of HBV infection and is universally present, equally strong in the USA and Europe as in Africa and Asia. The causal nature of the association between HBV and HCC appears indisputable, even though the pathogenesis of HBV-related HCC has not been established. There is no evidence to support any of the non-causal interpretations, i.e., that a third factor causes both persistence of HBV and HCC, or that HCC increases susceptibility to the HBV carrier state. Since about 200 million people are hepatitis B surface antigen carriers, HBV would appear to be second only to cigarette smoking as the most important known human carcinogen. However, a substantial proportion of HCC cases throughout the world show no evidence of active HBV infection; for those cases other causal agents must be invoked. Aflatoxin may be an important etiologic factor in Africa and Asia, whereas in Europe and the USA cigarette smoking and alcohol drinking are serious and numerically important suspects for hepatocellular carcinogenicity. Several other factors, including natural and synthetic chemical carcinogens, infectious agents and steroid hormones are also suspected, but the supporting evidence is weak and the numerical importance of the factors probably limited. PMID- 6295935 TI - Cytogenetics of heritability in cancer. AB - A possible causal association between chromosome structural change and neoplastic transformation has long been mooted, particularly since chromosomal changes occur frequently in the cells of a variety of malignancies. Only in recent years, however, has the evidence in support of this contention begun to appear convincing, and this has followed from the application of developments in cytogenetic techniques. The advent of methods for revealing specific bands in the human metaphase complement has enabled all the chromosomes and many chromosomal regions to be unambiguously identified, and the recent application of prophase banding methods gives further improvements in resolution. With these techniques, specific constitutional chromosomal deletions or translocations have been discovered in inherited cases of retinoblastoma (del.13q14), Wilms' tumour with aniridia (del.11p13) and renal-cell carcinoma (t(3:8) (p21:q24)), in which each of the chromosomal changes appears to be a dominant factor in inheriting a predisposition to a tissue-specific tumour. A heritability for cancer predisposition is also associated with the inherited chromosomal instability syndromes of Bloom's, Fanconi's anaemia and ataxia telangiectasia, although specific chromosomal changes have not been reported to be associated with the neoplasms in such individuals, except in some cases of lymphoma and leukaemia in ataxia telangiectasia. Specific chromosomal translocations have, however, been recorded in a variety of malignancies, with a particular involvement of chromosomes 22, 14, 8, 15, 17 and 21. However, although many hundreds of patients with the specific 9/22 rearrangement seen in chronic myeloid leukaemia and also those with the 14/8 rearrangement in Burkitt's, and other, lymphomas have been described, no single case in which these rearrangements were present as constitutional changes has been reported. The possible nature of the changes seen at the cytogenetic level in terms of gene content of the chromosomes involved is discussed. PMID- 6295936 TI - Complex, dynamically interacting host factors that affect the disposition of drugs and carcinogens. PMID- 6295937 TI - Carcinogenesis as a multistage process--experimental evidence. PMID- 6295938 TI - Laboratory decontamination and destruction of carcinogens in laboratory wastes: some N-nitrosamines. PMID- 6295939 TI - Effect of various pharmacological agonists on the rat caecum-appendix preparation: a preliminary study. AB - A preliminary study with various pharmacological agents revealed the presence of contractile muscarinic and H1-histamine receptors as well as inhibitory alpha- and beta-adrenoceptors and purinoceptors in rat caecum-appendix. Histamine also produced indirect actions mediated through the release of catecholamines. 5 Hydroxytryptamine produced variable response, the contractile response was mediated through 5-HT receptors and the relaxant response through the release of catecholamines. PMID- 6295940 TI - Effects of immunosuppression with cyclophosphamide on acute murine cytomegalovirus infection and virus-augmented natural killer cell activity. AB - The effects of cyclophosphamide (CY) treatment on acute murine cytomegalovirus (MCMV) infection were studied to explore the potential usefulness of MCMV as a means of detecting immune dysfunction and to identify host defense mechanisms important for protection against MCMV. Conditions found optimal for enhancing MCMV infection with CY included infecting adult mice with 2 X 10(5) PFU or more of virus and administering 80 mg or more of CY per kg 1 to 3 days later. In addition to enhanced mortality, virus titers in lung, liver, and spleen were elevated in CY-treated mice, and wet weights of liver, spleen, and thymus were depressed when compared with those of infected but untreated mice. Treatment with CY before MCMV challenge was not as efficient a means of enhancing mortality as treatment after virus challenge. The effect that the time of CY administration relative to infection had on mortality correlated with the effect of such timing on natural killer cell activity. Animals treated before infection exhibited depressed natural killer cell activity initially. However, they rapidly recovered this response, and by 5 days postinfection they had the same level of virus augmented activity seen in untreated mice. In contrast, animals treated after infection did not recover natural killer cell activity and were more likely to die. A similar correlation was not obtained when the effects of CY on lymphocyte responses to B and T cell mitogens were examined, nor were there striking differences in pathology between the treatment groups. The data suggest an important role for natural killer cells in host defense against MCMV. Also, increased susceptibility to MCMV may provide a useful indicator of deficits in the natural killer cell response. PMID- 6295941 TI - Detection of antibodies to pre-early nuclear antigen and immediate-early antigens in patients immunized with cytomegalovirus vaccine. AB - To define the role of antibody to immediate early antigens of human cytomegalovirus (CMV) in diagnosing acute infection, we studied antibody responses of adult volunteers and renal transplant candidates receiving CMV vaccine. In addition, CMV antibodies in healthy adults were determined. Antibody to pre-early nuclear antigen, one of the immediate early antigens, did not develop in all of the volunteers or renal transplant candidates receiving the vaccine. In those vaccinees who developed an antibody response, the duration of the response was variable. Antibody to pre-early nuclear antigen and immediate early antigens was also detected in some healthy adults with serological evidence of past CMV infection, but without clinical evidence of recent infection. In view of these results, antibody to immediate early antigens and specifically to pre early nuclear antigen does not appear to be suitable for rapid diagnosis of acute CMV infection. PMID- 6295942 TI - Surface receptors for serum albumin in group C and G streptococci show three different types of albumin specificity. AB - A total of 100 bacterial strains were tested for binding uptake of radiolabeled albumin preparations from 15 mammalian species. Three types of surface structures with specific binding sites for albumin were defined. A previously described receptor for albumin was separated into type a in Streptococcus equisimilis strains and in human group G streptococcal strains and type b in bovine group C streptococci. A new type of albumin receptor, type c, was found in Streptococcus dysgalactiae strains, the only receptor type so far with high affinity for bovine serum albumin. Type of albumin receptor correlated with bacterial species. The three receptor types showed high binding capacities; 2 X 10(8) bacterial organisms bound from 5 to 16 micrograms of albumin. All types of albumin receptors were stable to heat treatment at 80 degrees C for 5 min, but susceptible to both pepsin and trypsin treatment. Bacteria-bound albumin preparations were eluted at various concentrations of KSCN, reflecting differences in affinity. Up to 500 micrograms of human fibrinogen or polyclonal human immunoglobulin G had no inhibitory effect on the uptake of albumin, indicating a separate molecular localization of receptors for these proteins. PMID- 6295943 TI - Murine cellular cytotoxicity to syngeneic and xenogeneic herpes simplex virus infected cells. AB - Cellular cytotoxicity of C57BL/6 adult mice peritoneal cells to xenogeneic (Chang liver) and syngeneic (BL/6-WT3) herpes simplex virus (HSV)-infected cells was analyzed in a 6-h 51Cr release assay. There was no difference in antibody dependent cellular cytotoxicity to either target. There was no natural killer cytotoxicity to targets with cells from uninfected mice except at very high effector cell ratios. HSV-infected (2 X 10(4) PFU intraperitoneally 1 day previously) mice mediated significantly higher antibody-dependent cellular cytotoxicity and required less antibody (10(-5) versus 10(-2) dilution), fewer cells, and less time to kill than cells from uninfected mice. HSV-infected mice mediated natural killer cytotoxicity but preferentially killed syngeneic HSV infected cells. Stimulation of cytotoxicity was not virus specific since influenza-infected mice mediated similar levels of cytotoxicity to HSV-infected targets. There was no difference in morphology (95% macrophage) or in the percentage of FcR-positive cells, but infected mice had more peritoneal cells and generated higher levels of superoxide in response to opsonized zymosan or phorbolmyristate acetate. These data demonstrate nonspecific virus-stimulated metabolic and effector cell function which may enhance clearance of virus in an infected host. PMID- 6295944 TI - Interferon production by leukocytes infiltrating the lungs of mice during primary influenza virus infection. AB - Lung fluids and leukocytes were obtained from unprimed C3H mice by transpleural lavage at intervals after infection with influenza A/Hong Kong/68 virus and were tested for interferon activity. Lavage fluid interferon titers correlated directly with lung virus titers and with initial increases in leukocyte yields from infected lungs. In contrast to cultured lymph node cells from infected animals or leukocytes from lungs of uninfected mice, washed leukocytes obtained from the lungs of mice infected 2 to 6 days earlier produced interferon spontaneously in culture. The physiochemical, biological, and antigenic properties of both the interferon in lavage fluids and that produced by lung lavage leukocytes were similar and characteristics of alpha interferon. Fractionation studies indicated that macrophages and T lymphocytes were primarily responsible for the interferon produced in culture. The early presence and significant numbers of interferon-producing leukocytes in infected lungs suggests that these cells have an early role in defense against influenza virus infection. PMID- 6295945 TI - Effect of alphavirus infection on mouse embryos. AB - Seven strains and mutants of the alphaviruses, Semliki Forest virus and Sindbis virus, differed in their lethality for mouse embryos and their mothers. The A7 strain and the neurovirulence mutant M103 of Semliki Forest virus were selected for detailed comparison. A7 produced 100% lethality of mouse embryos but was avirulent for their mothers. M103 did not kill embryos or their mothers but did induce postnatal immunity. This immunity could be induced in utero or by suckling to an immune mother. Infectious virus could be recovered from the brain, blood, and embryos of mice infected with A7. Mice infected with M103 developed a viremia of similar titer, but virus could not be recovered from brain or embryos. Electron microscopy showed multiplying virus in trophoblast and fetal tissue after infection with A7, but no virus could be detected in such tissue after infection with M103. Cultures of ectoplacental cone trophoblast cells incubated in the presence of A7 or M103 showed multiplying A7 in the giant cells, but no M103. It is concluded that A7 can traverse both the blood-brain barrier and the placenta, whereas M103 can traverse neither. PMID- 6295946 TI - In vitro effect of hydrocortisone on the attachment and ingestion phases of immunoglobulin G- and complement component 3b-mediated phagocytosis by human neutrophils. AB - The fluorescence quenching method (FQ method) was used to investigate the effect of hydrocortisone on the attachment and ingestion phases of immunoglobulin G (IgG)- and complement component 3b (C3b)-mediated phagocytosis by human neutrophils (PMNs). The results were compared with metabolic activity (O2- release) of the phagocytes. When the PMNs were treated with 5 X 10(-5) M hydrocortisone or more, both IgG-mediated and C3b-mediated interactions decreased. The number of intracellular particles decreased as the total number of PMN-associated particles decreased, indicating an effect mainly on particle attachment. This was substantiated by the fact that pretreatment of the PMN with cytochalasin B resulted in a hydrocortisone dose-related decrease of interacting particles. The FQ method made it possible to quantify the stimulus-phagocyte interaction in relation to the metabolic response. Superoxide anion release decreased at the highest concentration of hydrocortisone used (5 X 10(-3) M), which merely reflected the decreased number of interacting particles. No reduction in metabolic activation was obtained when the superoxide anion release was correlated with the number of interacting yeast-IgG particles. The results indicate that hydrocortisone primarily affects the binding capacity of Fc and C3b receptors, resulting in decreased metabolic activation. The effector functions, e.g., ingestion and metabolic activation, were not affected by hydrocortisone in this study. PMID- 6295947 TI - Effects of a phagocytosis-stimulating factor on the phagocytic process of polymorphonuclear neutrophils. AB - The effect of phagocytosis-stimulating factor (PSF) on phagocytosis was studied in detail. PSF did not affect the Fc receptor-mediated phagocytosis, whereas PSF enhanced the ingestion step, but not attachment step, of C3b receptor-mediated phagocytosis by polymorphonuclear neutrophils (PMNs), suggesting that PSF may specifically modulate the C3b receptor function of PMNs. PSF generated from guinea pig PMNs enhanced phagocytosis by rabbit PMNs, and rabbit PMN-produced PSF accelerated the phagocytosis by guinea pig PMNs, indicating that PSF is not specific for animal species. The effects of PSF on some PMN functions, such as O2 generation, chemotaxis, adherence, and enzyme release, were also studied. Only O2- generation from PMNs was significantly increased in the initial phase of phagocytosis, and this stimulation of O2- generation was completely parallel with the stimulation of phagocytosis by PSF. Resting PMNs hardly generated the superoxide anions by PSF treatment, suggesting that PSF does not affect the O2- forming system directly. PMID- 6295948 TI - Restriction endonuclease analysis of DNA from genital isolates of herpes simplex virus type 2. AB - High-resolution restriction endonuclease analysis of DNA from multiple herpes simplex virus type 2 isolates from 30 patients at a single clinical center has indicated that herpes simplex virus type 2 DNA sequences are highly conserved. However, the use of six endonucleases, EcoRI, BglII, HindIII, KpnI, BamHI, and SstI, established that each patient was infected by a unique virus. Comparison of virus isolates from the same patient show that the most sequence variation occurred in the terminal and subterminal BamHI fragments. The results suggest that each individual may induce specific variation in the herpes simplex virus type 2 genome and that the results of epidemiological studies based on restriction endonuclease analyses of herpes simplex virus type 2 DNA must be interpreted with care. PMID- 6295950 TI - Selective cytotoxicity of Clostridium perfringens delta toxin on rabbit leukocytes. AB - Clostridium perfringens delta toxin was selectively cytotoxic for various rabbit leukocyte populations. The sensitivity of these populations to the toxin varied, depending on the tissue from which they were derived, from 28% (appendix) to 41% (bone marrow) and 32% (spleen). Macrophages were uniformly killed by delta toxin, whereas thymocytes were essentially resistant. Selective cytotoxicity was correlated to the specific binding of the radiolabeled toxin by target cells. The relationship between sensitivity to the toxin and the presence of GM2 ganglioside in the cell membrane of rabbit leukocytes is discussed. Delta toxin might prove a useful new tool for separating leukocyte subpopulations based on their differential sensitivity to the cytolethal effect of this protein. PMID- 6295949 TI - Polymorphonuclear leukocyte activation by a synthetic muramyl dipeptide analog. AB - N alpha-(N-Acetylmuramyl-L-alanyl-D-isoglutaminyl)-N epsilon -stearoyl-L-lysine, a synthetic derivative of muramyl dipeptide, stimulated the chemotactic mobility, phagocytic activity, and superoxide anion (O2-) productivity of peritoneal polymorphonuclear cells in mice. The chemotactic mobility of both cells preincubated with the adjuvant in vitro and those derived from the mice previously treated with the adjuvant was significantly enhanced. The phagocytic activity of cells preincubated in vitro with the adjuvant was also enhanced transiently, and that of the cells derived from the mice treated subcutaneously with the adjuvant 24 h before intraperitoneal inoculation with Escherichia coli was significantly greater than that of cells from the mice given phosphate buffered saline instead of the adjuvant. The release of O2- from the cells derived from the adjuvant-treated mice was also greater than that from the cells of untreated control mice. However, the exposure of the cells derived from untreated mice to the adjuvant in vitro did not stimulate O2- generation by the cells. PMID- 6295951 TI - Comparison of the calcium ionophore and phorbol myristate acetate on the initiation of the respiratory burst in human neutrophils. AB - We investigated the ability of the calcium ionophore A23187 and phorbol myristate acetate to elicit a respiratory burst in human neutrophils. In general, the ionophore was considerably more potent than phorbol myristate acetate in generating a chemiluminescent response and slightly less active in generating superoxide anion. In contrast, the ionophore caused a much smaller stimulation of glucose oxidation via the hexose monophosphate shunt. This relative inability of the ionophore to stimulate the shunt could not be ascribed to an effect on glucose transport or to a direct inhibition of any of the enzymes involved in glucose metabolism. These data suggest that different stimuli may have markedly different effects on various activities associated with the respiratory burst and emphasize the necessity for measurement of more than one parameter to assess the oxidative metabolism of the neutrophil. PMID- 6295952 TI - Cytomegalovirus-infected cell polypeptides immune-precipitated by sera from children with congenital and perinatal infections. AB - Congenital or perinatally acquired human cytomegalovirus (CMV) infections in children may be symptomatic or asymptomatic. In this study, we characterized the electrophoretic properties of CMV-infected cell polypeptides immune-precipitated by sera from children with different types of CMV infections from birth to 4 years of age. Sodium dodecyl sulfate-polyacrylamide gel analysis of immune precipitates formed with radiolabeled extracts of cells infected with CMV strain AD169 showed the following. (i) Electrophoretic profiles of CMV polypeptides immune-precipitated by sera from children with perinatal and congenital infections were similar. At least 11 polypeptides with apparent molecular weights of 150,000, 140,000, 110,000, 100,000, 74,000, 66,000, 50,000, 49,000, 34,000, 25,000, and 20,000 were precipitated. Antibody titer in anticomplement immunofluorescence tests and virus titer in urine correlated with the intensity of polypeptide profiles in autoradiograms. (ii) The initial immune response of children with symptomatic congenital infections was delayed as compared to that of children with asymptomatic congenital and perinatal CMV infections. Sera obtained serially from symptomatic children for years after birth continued to precipitate CMV polypeptides, whereas sera from children with subclinical congenital infections precipitated lesser amounts over time. (iii) Immune precipitates obtained with sera from CMV-infected patients and with monoclonal antibodies to CMV contained polypeptides with comparable electrophoretic and immunological properties. PMID- 6295953 TI - Receptor-like glycocompounds in human milk that inhibit classical and El Tor Vibrio cholerae cell adherence (hemagglutination). AB - The two biotypes of Vibrio cholerae were found to have cell-associated hemagglutinins which differ with regard to binding to different species of erythrocytes and inhibition by monosaccharides. A total of 12 classical V. cholerae strains (Inaba or Ogawa) strongly agglutinated human erythrocytes in a reaction specifically inhibited by L-fucose, whereas 12 El Tor strains preferably agglutinated chicken erythrocytes, a reaction reversed by D-mannose or by higher concentrations of D-fructose, D-glucose, alpha-methyl-D-mannoside, or sucrose. Milk from Swedish women inhibited both of these adherence reactions, and the predominating inhibitory activity for each reaction resisted boiling, was destroyed by periodate treatment, and bound a concanavalin A-Sepharose column, suggesting a carbohydrate structure. Further characterization indicated that the inhibitory activity for classical V. cholerae hemagglutination was distributed about equally on glycoprotein and free oligosaccharide, but was not present on glycolipid. The El Tor inhibiting activity, on the other hand, was almost exclusively of a high-molecular-weight glycoprotein nature. These results support our previous suggestion (Holmgren et al., Infect. Immun. 33:136-141, 1981) that human milk may contain receptor-like glycocompounds which can prevent bacterial adherence by competition with receptors on target cells. PMID- 6295954 TI - Kinetics and genetics of herpes simplex virus-induced antibody formation in mice. AB - The kinetics of antibody synthesis was investigated after intraperitoneal, subcutaneous, and footpad infection of various strains of mice with herpes simplex virus. Immunoglobulin M antibodies appeared 5 days after and immunoglobulin G antibodies appeared 10 to 12 days after intraperitoneal infection with herpes simplex virus type 1. The major histocompatibility complex and the background genome of inbred mice were not found to have a systematical influence on antibody synthesis. Female mice, however, consistently produced more antibodies than did male if the infection was done intraperitoneally, but not if it was done subcutaneously or into footpads. Castration considerably increased the amount of antibodies produced by male mice. The difference in antibody formation between females and males could be abolished by injection of silica; moreover, antibody titers were enhanced by this treatment. This has also been found by immunization with a Formalin-inactivated herpes simplex virus vaccine. The effect of silica in enhancing antibody formation could be observed up to 12 days after infection. Infectious virus could be detected up to 2 days after infection, and herpes simplex virus type 1 antibody-stimulating antigens could be detected up to 4 days in ultrasonicates of macrophages. The assumption is made that androgen-sensitive cell populations, including macrophages and their soluble products, are involved in antibody-depressing mechanisms. PMID- 6295955 TI - Purification of an outer capsid glycoprotein of neonatal calf diarrhea virus and preparation of its antisera. AB - An outer capsid glycoprotein, VP 7, was purified from a bovine rotavirus, neonatal calf diarrhea virus, by isoelectric focusing in glycerol gradients after disruption of the outer capsid. Its isoelectric point was found to be about 4.5. Guinea pigs were immunized with this VP 7 preparation. The antisera possessed both neutralizing and hemagglutination-inhibiting activities as well as complement-fixing activity, suggesting that VP 7 is a protein involved in hemagglutination and initiation of infection. When these antisera were reacted with a simian rotavirus, their antibody titers were low by hemagglutination inhibiting and complement-fixing assays, but one serum neutralized the simian rotavirus as efficiently as it did homologous neonatal calf diarrhea virus. PMID- 6295956 TI - Passive protection of mice and sheep against bluetongue virus by a neutralizing monoclonal antibody. AB - A murine hybridoma antibody, 6C2A.4.2, previously characterized as an immunoglobulin G class 2a that binds in radioimmunoassay to bluetongue virus serotype 17 (BTV-17) but not the other 19 BTV serotypes, neutralizes BTV-17, inhibits hemagglutination with BTV-17, and precipitates viral polypeptides 2 and 3 from BTV-17-infected cells, was produced as an ascites in the peritoneal cavities of hybridoma-inoculated mice. This ascitic fluid, but not those containing other, non-neutralizing anti-BTV-17 antibodies of the same isotype, provided serotype-specific passive protection against BTV-17-induced death of neonatal mice. Antibody 6C2A.4.2-containing ascitic fluid was injected intravenously into sheep that were later inoculated with BTV-17. These sheep remained free of clinical signs, did not develop viremia or detectable levels of antibodies reactive in the immunodiffusion test used for routine BTV diagnosis in the United States, and developed only low levels of neutralizing antibodies. Control animals became viremic and developed immunodiffusion test reactions and high levels of neutralizing antibodies during recovery, and two of three had lesions and fevers. These results provide evidence that antibodies directed against a single epitope on BTV-17 can prevent bluetongue disease. PMID- 6295957 TI - Cell-mediated immunity against herpes simplex virus envelope, capsid, excreted, and crude antigens. AB - Cell-mediated immunity to herpes simplex virus envelope, capsid, excreted, and crude antigens was studied by in vitro lymphocyte stimulation tests during 198 recurrent attacks in 69 patients. Excreted antigen caused no blast transformation. Envelope and capsid antigen-induced lymphocyte stimulation was at the maximum 7 to 14 days postinfection, declining thereafter to a rather constant level in 1 to 2 months. The lowest levels were measured just a few days before a new attack. In persons with frequent relapses, the fluctuation was more rapid and stimulation index levels stayed higher, although no protective level seemed to exist. Cultures stimulated with the crude antigen in autologous serum showed rapid increases and declines in the stimulation index values, contrary to those grown in agamma serum, in which the stimulation level stayed rather constant up to 1 year postinfection. PMID- 6295958 TI - Shigella dysenteriae 1 cytotoxin: periplasmic protein releasable by polymyxin B and osmotic shock. AB - Treatment of Shigella dysenteriae 1 either with the antibiotic polymyxin B or by osmotic shock resulted in the release of 80 to 90% of the cytotoxin activity of the organism. Under the conditions employed, the release of toxin activity was accompanied by the appearance of a periplasmic enzyme, 5'-nucleotidase. There was no significant release of cytoplasmic contents, assessed by measurement of glucose-6-phosphate dehydrogenase activity. The release of cytotoxin and 5' nucleotidase by polymyxin B were both dependent on the duration of incubation with, and the concentration of, the antibiotic. In terms of specific activity (cytotoxin activity per milligram of protein), the polymyxin B and osmotic shock extracts were 20- to 30-fold more active than crude toxin preparation derived from a whole-cell lysate. The data strongly support a periplasmic location for Shiga cytotoxin and the utility of the polymyxin B extraction to obtain starting material for toxin purification. PMID- 6295959 TI - Rous-associated virus type 7 induces a syndrome in chickens characterized by stunting and obesity. AB - Infection of 10-day-old chicken embryos with an avian retrovirus. Rous-associated virus type 7, resulted in a disease characterized by stunting and hyperlipidemia. By 20 days after hatch, infected chickens were smaller than hatchmates and developed ataxia and obesity over the next 30 days. Histological examinations of livers from infected chickens revealed a diffuse panlobular fatty infiltrate involving an accumulation of fat in microdroplets. Electron microscopic examinations of livers from infected chickens revealed hepatocytes with swollen mitochondria that lacked cristae. The thyroid and pancreas were infiltrated with lymphoblastoid cells by 1 week after hatch. An examination of the blood revealed a mild anemia, a frank lipemia, and high levels of uric acid. This syndrome induced by Rous-associated virus type 7 in chickens may be useful for elucidating the nature of several diseases, including that found in the fatty liver and kidney syndrome of chickens and that observed in a strain of obese chickens. PMID- 6295960 TI - Effect of oxygen-dependent antimicrobial systems on Legionella pneumophila. AB - Legionella pneumophila was susceptible to the antimicrobial action of oxygen metabolites generated by both the myeloperoxidase-H(2)O(2)-halide and the xanthine oxidase systems. PMID- 6295962 TI - [Complications in blood transfusion. 2. Infections transmitted by blood and blood products]. PMID- 6295961 TI - p105, an Epstein-Barr virus-induced, phosphonoacetic acid-insensitive glycoprotein target of the anti-Epstein-Barr virus immune response. AB - To describe structures and biological functions of targets for antibody-mediated immune responses to Epstein-Barr virus (EBV)-infected lymphoid cells, we have characterized a membrane-associated protein of 105,000 daltons, p105, which was prominently recognized in immunoprecipitates with some EBV antigen-reactive patients' sera. A rabbit antiserum to immunopurified p105 was developed. [35S]methionine-labeled p105 was specific to EBV-superinfected Raji cells, and its synthesis was not blocked with phosphonoacetic acid, indicating that it is an "early" viral antigen. Phosphonoacetic acid treatment of EBV-superinfected Raji cells was associated with the accumulation, mainly in the cytosol, of an 88,000 dalton protein, p88, which was also recognized with anti-p105 serum, but was not detected in superinfected cells which had not been treated with phosphonoacetic acid. Although anti-p105 serum immunoprecipitated a membrane fraction protein, it did not neutralize P3HR-1 virus and was not considered to be an exposed virion component. We conclude that p105 is an early, EBV-induced, membrane fraction antigen to which EBV-infected patients generate a substantial antibody response. PMID- 6295963 TI - Pharmacologically induced changes in human beta-chorionic gonadotropin (beta-hCG) synthesis by human placenta in organ culture. AB - In organ cultures the regulation of beta-hCG formation in human placentas of early gestation (EG) and at term (T) was analyzed using the following test substances: dibutyryl cAMP (dbcAMP), theophylline, and by two drugs used to treat patients with premature labor, fenoterol hydrobromide and verapamil hydrochloride. The rapid increase in beta-hCG in culture medium beginning at 24 48 h corresponds to morphologic changes (proliferation of the cytotrophoblast). Beta-hCG synthesis of human placenta is related to an adenylate cyclase as could be shown by dose-dependent stimulation by dbcAMP (T) (after 72 h) (controls = 100%): 2 nM = 125%; 70 nM = 200%; 2 microM = 300%. Similar results were obtained using EG. First stimulation by dbcAMP was found after 2 h and may be caused by a release of intracellular beta-CG or by a direct stimulation of beta-hCG synthesis; late effects (after 48-72 h) are suspected to be due to changes in morphologic differentiation of the placenta in organ culture. Theophylline (0.1 and 1 mM) did not influence beta-hCG formation in EG, whereas in T a dose dependent stimulation could be achieved (controls = 100%): 0.1 mM = 110%; 1 mM = 300%; and 10 mM = 160%. Fenoterol hydrobromide, a tocolytic beta 2-receptor stimulating agent showed a slight (130-160%) beta-hCG stimulation (after 72 h) (EG and T). The stimulatory effect of a high dose (1 microgram/ml) of fenoterol hydrobromide could be due to a stimulation of beta 2 receptors related to an adenylate cyclase. Dose-related changes in beta-hCG in T were found for the calcium antagonist verapamil hydrochloride (controls = 100%): 0.01 microgram/ml = 125% stimulation; 0.33 microgram/ml = 200% stimulation; 10 micrograms/ml = 100%. As indicated by verapamil tests, calcium ions may play a role in the regulation of beta-hCG production. PMID- 6295964 TI - Antibody responses to Epstein-Barr virus-specific DNase in relation to the prognosis of juvenile patients with nasopharyngeal carcinoma. AB - We have examined serial sera from 17 juvenile patients with nasopharyngeal carcinoma (NPC) for their capacity to neutralize the activity of Epstein-Barr virus (EBV)-specific DNase. The results revealed that NPC patients who became long-term survivors (LTS) without evidence of the disease either never possessed significant levels of antibodies to the enzyme or showed a gradual decline in the number of EBV DNase units neutralized from an elevated level at diagnosis to an insignificant figure several years later. All the 10 LTS neutralized less than 4, and some neutralized less than 2 units of the enzyme 3 or more years after the initial diagnosis. In contrast, serial sera from juvenile patients who died of NPC neutralized over 10 and as many as 25 units of EBV DNase either persistently until death occurred or with transient declines during unmaintained remissions. Rises and declines in the neutralizing activity were, with few exceptions, accompanied by corresponding changes in the titers of IgA and IgG antibodies to EB viral capsid antigen and to the diffuse component of the early antigens. Although the number of juvenile NPC cases available for study was small, the observations suggest that the EBV DNase neutralization test may serve to provide information on the prognosis of the patients. PMID- 6295965 TI - Human embryonal carcinoma cells in culture do not synthesize fibronectin until they differentiate. AB - The expression of fibronectin by clones of 2102Ep, a human cell line derived from a testicular germ-cell tumor, was studied. In high-density cultures these cells retain an embryonal carcinoma phenotype and do not express the cell surface antigen SSEA-1. They do not synthesize fibronectin. However, when grown at low densities, many of the cells appear to differentiate, as indicated by a change in their morphology and their expression of SSEA-1. Such cultures also synthesize fibronectin. Further, when low-density cultures were fractionated into SSEA-1 positive and SSEA-1-negative subpopulations by fluorescence-activated cell sorting, fibronectin synthesis was confined to the SSEA-1-positive cells. The fibronectin produced by these cells exhibits an apparent molecular weight (261,000) slightly greater than that produced by diploid fibroblasts (250,000) and may represent an embryonic form. It is not laid down as an extracellular matrix and cannot be detected by immunofluorescence analysis of cell monolayers. PMID- 6295966 TI - Discordant Epstein-Barr virus nuclear antigen (EBNA) antibody patterns in nasopharyngeal carcinoma. AB - Epstein-Barr virus nuclear antigen (EBNA) preparations from three sources were tested with sera from normal individuals and patients with Hodgkin's disease, breast carcinoma, Burkitt's lymphoma, and American and Chinese nasopharyngeal carcinoma. Individual sera with discordant antibody patterns were noted in all groups. Sera from both NPC groups gave significantly higher anti-EBNA titers on cell lines converted with P3HR-1 or B95-8 virus compared with anti-EBNA titers on Raji cells. Anti-EBNA titers of Chinese NPC sera showed no correlation among the three EBNA sources, while all other groups had highly correlated titers. Cross absorption experiments present evidence for more than one antigenic determinant on EBNA. These results suggest an additional parameter for distinguishing Chinese NPC from other EBV-related disorders. PMID- 6295967 TI - Hybridization between a human epithelial line, infectable by Epstein-Barr virus, and Burkitt lymphoma lines: membrane properties, superinfectability, inducibility and tumorigenicity. AB - The human epithelial line U, which is partially infectable with EBV, was hybridized with the EBV-genome carrying Burkitt lymphoma lines P3HR-1 and Daudi. Authenticity of the hybrids U-Put and U-Dut was established by isoenzyme studies. Although the two hybrids carried the EBV genome derived from the lymphoma parent, being 100% positive for Epstein-Barr-virus-associated nuclear antigen (EBNA), they resembled the U parent in many respects: they were deficient for membrane immunoglobulins and Fc receptors, and had a lower concentration of EBV-C3 receptors than either parent. Unlike the P3HR-1 parent, U-Put hybrid was nonpermissive for both the EBV cycle antigens, early antigen (EA) and viral capsid antigen (VCA). The inducing agent 12-O-tetra-decanoyl-phorbol-13-acetate (TPA) caused distinct viral early antigen synthesis (EA) in U-Put, lower, however, than that of the parental P3HR-1. U-Dut was completely nonpermissive and noninducible for early and viral capsid antigens. Thus, even an epithelial parent infectable by EBV restricted, although not completely, expression of EBV antigens, with the exception of EBNA. It has been suggested that EBNA is an autonomous function of the viral genome, independent of host cell control; the latter regulates expression of antigens related to viral cycle. The hybrids U-Put and U-Dut resembled the U parent also in regard to growth in soft agar and tumorigenicity in nude mice, although in this respect the lymphoma parental properties were not completely eclipsed. PMID- 6295968 TI - Tumorigenicity of tsA and wild-type simian virus 40 transformed cells inoculated onto the chicken chorioallantoic membrane. AB - This study compares the tumorigenicity of SV40 primary tumor cell lines, tsA and wild-type SV40-transformed Chinese hamster cells, at two temperatures, 37 degrees C and 40.5 degrees C, inoculated onto the chorioallantoic membrane of the chicken egg. The SV40 primary tumor cell lines varied in their efficiency of takes at 37 degrees C from 78% for the H65-90B tumor line, 73% for the H80-7A and 25% for the H80-4 line. At 40.5 degrees C the H80-4 was unable to form tumors; however, the H80-7A and H80-4 produced 70% and 20% tumors respectively. Histologically, all CAM tumors were fibrosarcomas identical to the transplanted tumors, however, the tumor(s) at 40.5 degrees C were smaller. Chinese hamster wild-type SV40 transformed cells were equally efficient (32%) at tumor production at both temperatures. The tsA-SV40-transformed Chinese hamster cells (A58 and A58-2) induced 34% tumors at 37 degrees C and 9% tumors at 40.5 degrees C. At 37 degrees C these tumors were typical fibrosarcomas; however, the 40.5 degrees C tumors were smaller and less cellular, resembling a more differentiated fibrosarcoma. Therefore, the tsA Chinese hamster transformed cells were less efficient at tumor induction at the non-permissive temperature; however, the primary tumor lines also demonstrated a variability in tumorigenicity (H65-90B and H80-4). Possibly factors other than the temperature-sensitive viral protein (big "T" antigen) may be involved in establishing a tumor on the chicken CAM. PMID- 6295969 TI - Changes in antibody titers in cattle infected clinically and subclinically with bovine leukemia virus. AB - Changes in titers of antibodies to protein and glycoprotein antigens of bovine leukemia virus (BLV) were investigated by immunodiffusion in two groups of cattle. In one group of cattle (10 head) which developed enzootic bovine leukosis during the observation period of 3 years, seven showed a gradual increase in antibody titers following a period of level antibody titers that persisted for several months to several years. The titer was finally eight to 32 times higher than the initial titer. The remaining three showed little change in titer. Of the 10 animals with tumors, three showed a decrease in the titer when they were in extremis. In contrast, in the second group of cattle (110 head), which were infected with BLV subclinically, the change in the titers during 1 and 3 years ranged from no change to four times the lowest titer that was observed during the period. Of the cattle which had a low level of antibody to one of the BLV antigens, a few became negative reactors to the respective antigen. PMID- 6295970 TI - Secretion of basement membrane collagen degrading enzyme and plasminogen activator by transformed cells--role in metastasis. AB - The relationship of a basement membrane collagen degrading enzyme (BM collagenase) and plasminogen activator (PA) was studied in a number of non malignant and malignant human and murine cell lines. Several non-malignant cell lines secreted significant amounts of PA but not detectable BM collagenase activity whereas the malignant cell lines, with one exception, secreted both enzymes. Therefore, the secretion of BM collagenase appears to be a characteristic of many malignant cells whereas PA is synthesized also by normal cells. The BM collagenase needed proteolytic activation for maximal activity indicating that it is secreted in a latent form. The addition of plasminogen to the culture medium of human fibrosarcoma cells (HT-1080) resulted in maximal activation of the enzyme. Plasmin, but not plasminogen, increased the activity of partially purified enzyme protein. Accordingly, the activation of latent BM collagenase in vivo may be facilitated by PA through the conversion of plasminogen to plasmin. It is suggested that the secretion of BM collagenase concomitantly with PA is a prerequisite for metastasis. PMID- 6295971 TI - The renin-angiotensin system. A primary factor in the control of coronary flow? PMID- 6295972 TI - Effects of chronic administration of bronchodilators on microshock in sensitized guinea-pigs. AB - In conscious guinea-pigs isoprenaline, adrenaline and salbutamol protected the animals against egg albumin and histamine microshock in a dose-dependent manner. Chronic aerosol pretreatment three times daily with adrenaline or isoprenaline, but not with salbutamol, enhanced the acute dose protection against egg albumin microshock, but there was no enhancement when the bronchodilators were given six times daily. Aminophylline when administered three times daily enhanced the protective effect of an acute dose of adrenaline. With histamine microshock, desensitization occurred to adrenaline, when given three and six times daily, and to isoprenaline, when given six times daily, but not to salbutamol. Cross desensitization could be induced to adrenaline by isoprenaline but not by salbutamol. The data indicate that, depending on the experimental conditions, enhancement of or desensitization to the effects of bronchodilators could be shown in conscious guinea-pigs. PMID- 6295973 TI - Ketoconazole: a new imidazole antifungal agent has both prophylactic potential and therapeutic efficacy in keratomycosis of rabbits. AB - A highly reproducible quantitative model of fungal infection of the rabbit's corneal stromal was produced using multiple corneal microtrephination. Aspergillus flavus (K4/77), at a concentration of 10(6) spores per ml was systematically implanted into the trephine sites in the cornea, and the degree of corneal infection determined. By pre-inoculation and post-inoculation challenge of these cornea with 1% ketoconazole in arachis oil, the prophylactic potential and the therapeutic usefulness of ketoconazole was determined. Ketoconazole, acetyl-dichlorophenyl-imidazole, has a significant prophylactic potential in inhibiting the development of corneal stromal fungal lesions when it is administered to the cornea of New Zealand white male rabbits as a 1% solution in arachis oil for two consecutive times hourly for two hours before the inoculation of the rabbits cornea with an ocular pathogenic Aspergillus flavus. Ketoconazole also has a therapeutic effect in the reduction of well established A. flavus keratitis in rabbits. When administered as 1% solution in arachis oil for ten consecutive hours daily to well established A. flavus lesions of the cornea of New Zealand albino rabbits, ketoconazole took about sixteen days to cure all the corneal lesions. Finally, using a yeast nitrogen base liquid medium, the in vitro minimal inhibitory concentrations of ketoconazole to twenty-five various human ocular pathogenic fungal isolates were determined and used to recommend those fungi for which ketoconazole would be a good choice for therapy. PMID- 6295974 TI - Study of ketoconazole toxicity in rabbit cornea and conjuctiva. AB - Ketoconazole, acetyl-dichlorophenyl-imidazole, when administered topically to the eyes of normal New Zealand white rabbits in concentrations of 1%, 3% and 5% in arachis oil has been shown to have no toxic effect to the conjunctival and corneal epithelium of these animals. No toxic effects were also noted on the iris sphincter and the crystalline lens of these animals. Five per cent ketoconazole in arachis oil showed some minimal conjunctival hyperaemia in two of the test eyes (33.3%) after two weeks of continuous administration. Arachis oil, the carrier for the test drug showed no ocular toxicity in all the rabbits tested. PMID- 6295975 TI - Radiosensitization of single-stranded phi X 174 DNA in aqueous solution by misonidazole is induced by metalloporphyrins. PMID- 6295976 TI - The relation of programmed cell death to development and reproduction: comparative studies and an attempt at classification. PMID- 6295977 TI - Motility during fertilization. PMID- 6295978 TI - Interaction of viruses with cell surface receptors. PMID- 6295979 TI - Effect of particle size on the tissue distribution of iodized emulsified fat following intravenous administration. AB - Three iodinated fat emulsions were tested by intravenous injection to rats: coarse with a mean size of particles of 7 micrometers, fine with a mean size of 1.3 micrometers, and ultrafine with a mean size of 0.7 micrometers. Iodine content analysis and scintigraphy of radioactive iodinated emulsions show important differences. Coarse emulsion is almost entirely fixed in the liver and the lungs; fine emulsion has a higher relative liver fixation; and ultrafine emulsion has a hepatosplenic fixation of short duration. Degradation of contrast medium is slow with coarse emulsion and very fast with ultrafine emulsion which is responsible for the rising iodine concentration in blood. Fine emulsion is a good compromise between excessive pulmonary fixation of coarse emulsion and high iodine peak in the blood after injections of ultrafine emulsion. The dose of 0.2 ml/kg of fine emulsion gives persistent enhancement of hepatic parenchyma without significant toxicity. PMID- 6295980 TI - Malignant fibrous histiocytoma arising from an exostosis in a patient affected by multiple exostoses. PMID- 6295981 TI - Further studies on the absorption of actinide elements from the gastrointestinal tract of neonatal animals. AB - Plutonium retention was measured after intragastric administration to neonatal rats, dogs and swine. At 1 week after administration, substantially more of the actinide remained in swine and dogs than in rats. The quantity of 238Pu absorbed by piglets was markedly influenced by such factors as compound solubility, mass of plutonium administered, oxidation state of the actinide, and age of the animal at gavage. Cortisone treatment reduced absorption, but was less effective in piglets than in neonatal rats. Measurements of 238Pu transport from ligated segments of the neonatal swine intestine indicated highest absorption from the duodenum, where the actinide was shown, autoradiographically, to be deposited in the epithelial region; in the ileum, deposition was predominantly in the lacteal region. Absorption of actinides by neonatal swine decreased in the order of 233U greater than 238Pu greater than 237Np greater than 244Cm greater than 241Am. Measurements at 1 yr after gavage showed a much higher retention by swine than by rats. PMID- 6295982 TI - Radon and its daughters in European mines. PMID- 6295983 TI - A study on the radiological characteristics of Norwegian non-uranium mine atmospheres. PMID- 6295984 TI - [Diagnostic value of neuropsychiatric, audiological and electrophysiological examinations of alcohol-dependent patients]. AB - Neuropsychiatric, audiological and electrophysiological examinations performed on 52 patients under neuropsychiatric therapy after long-term alcohol dependence, gave evidence of polytopical impairment of the peripheral and central nervous systems as well as inner ear. The results show that in order to ascertain impairments, a comprehensive diagnostic program is to be applied with special focus on electroneurographic examinations of peripheral nerves. According to the present findings, the latter are also recommended as a screening method, in particular scrutinizing the N. fibularis while also determining the threshold audiograms. PMID- 6295985 TI - Multimodality treatment of patients with advanced ovarian carcinoma. AB - A multimodality treatment program has been applied to ovarian carcinoma at the Johns Hopkins Hospital since August 1975. Forty-nine patients were subdivided into 23 patients with maximally resected Stage III micrometastatic, and 26 patients with significant retained disease, 20 with Stage III macrometastatic and 6 with Stage IV. After initial pilot studies, those patients with minimally retained disease entered a randomized prospective study. Antiovarian antiserum was used in one arm of the study; in both study arms colloidal P-32, delayed split whole abdominal irradiation, and maintenance melphalan were used. For the 23 patients with micrometastatic disease the cumulative survival and survival without evidence of disease at four years is 78 and 34% respectively. Twenty-six patients with macrometastatic disease were treated with or without intraperitoneal antiserum and multiagent chemotherapy; their cumulative one year survival is 50%. The lack of significant toxicity of intraperitoneal antiovarian antiserum and the results of multimodality therapy indicate the feasibility of this therapeutic approach to further improve ovarian cancer therapy. PMID- 6295986 TI - Superfractionation in glioblastoma multiforme--results of a phase II study. AB - A sequential series of 30 patients who were referred to a cancer treatment hospital with glioblastoma multiforme were treated with superfractionated cobalt 60 gamma radiation, three treatments per day, 100 rad per fraction. Their survival was compared to that of a historical group of 90 patients who had been referred for the same disease. Survival of the study patients was significantly longer than the historical patients, both for those who underwent resection (48.6 weeks median survival vs. 35.1 weeks), and for those who did not (35.1 weeks vs. 11.7 weeks). A retrospective survey of the historical group led to the following conclusions about this group: 1. Survival for patients who were well enough to be referred was unchanged after steroids came into general use; 2. Younger patients (under 50 years) did not have a longer survival than older patients; 3. The size of the dose of irradiation did not affect survival over the range of doses employed; 4. The size of the treatment volume employed did not affect survival over the range of treatment volumes employed. PMID- 6295987 TI - Critique of "Primary malignant cerebellar astrocytomas in children: a signal for postoperative cranio-spinal irradiation". PMID- 6295988 TI - The in vivo fate of a 211At labelled monoclonal antibody with known specificity in a murine system. AB - A monoclonal antibody reactive against the human transferrin receptor has been labelled with the alpha and X ray emitting isotope Astatine 211. The labelling procedure does not affect the ability of the product to bind to the transferrin receptor on the human leukemic cell line HL60. Using a direct binding assay, 211At labelled antibody can be specifically inhibited from binding to its target cells by excess unlabelled antibody. Furthermore, the binding inhibition demonstrated in this system correlates to enhanced clonogenic survival of these cells, indicating that very few atoms of 211At/cell are required for cell death. Data obtained from labelled antibody injected into mice show that the labelled product in serum retains the ability to bind to HL60 cells in vitro, although tissue distributions of the injected activity implies that some of the radiolabel is lost from the protein. Despite this loss of label, preliminary experiments on the localization of labelled antibody to HL60 cells growing s/c in nude mice show that tumor tissue has a higher specific activity than all other tissues, other than blood, after 12 hours. This suggests that further work on the nature of label degradation in vivo is warranted in the context of potential therapeutic and diagnostic studies. PMID- 6295989 TI - Influence of exercise intensity and duration on biochemical adaptations in skeletal muscle. AB - The influence of intensity and daily duration of exercise on cytochrome c concentration in the three muscle fiber types was assessed in rats that were treadmill trained for 8 wk (5 days/wk) by 1 of 19 protocols. The importance of exercise duration was determined at six running intensities (10, 20, 30, 40, 50, and 60 m/min). Daily run times resulted in a maximal adaptive change for each running intensity. The general pattern of cytochrome c change, caused by increasing daily run times, followed a first-order manner in all fiber types. The final time-independent asymptotic values were dependent on the intensity of running. In addition, as running intensity was increased, the length of daily run time required to achieve the peak adaptive response was shortened. The intensity influence was very different between fiber types. In the fast-twitch red fiber, the maximal time-independent response at each work intensity was altered by exercise intensity at only submaximal work loads (i.e., the adaptation from 19 to 29 nmol/g was not further increased at 50 and 60 m/min). In contrast, the adaptive response in the fast-twitch white fiber began at 30 m/min (approx 80% VO2max) and increased exponentially as intensity was increased (200% increase at 60 m/min). Whether this different adaptive response between fiber types is due solely to the ordered recruitment of motor units remains to be determined. PMID- 6295990 TI - Homogeneous nucleation of gas bubbles in vivo. AB - Several current theories of decompression sickness (DCS) presume the preexistence of gas bubble nuclei in tissue, because the de novo nucleation of gas bubbles in the body is thought to be theoretically impossible. Reexamination of nucleation theory reveals the overwhelming importance of two parameters: gas supersaturation and tissue surface tension (gamma). For the high gamma of pure water nucleation theoretically requires more than 1,000 ATA supersaturation. Lower values of gamma allow nucleation to occur with vastly smaller supersaturations. Application of homogeneous nucleation theory can provide reasonable fits to both rat and human pressure-reduction data with values of gamma within the range reported for biological fluids (below 5 dyn/cm). The initial bubble sizes predicted are 0.1 micron or less. The presence of heterogeneous sites, for example crevices and lipid surfaces, makes nucleation even more likely. PMID- 6295991 TI - Comparative brain oxygenation and mitochondrial redox activity in turtles and rats. AB - Comparisons were made between brain oxygenation and cytochrome c oxidase (cytochrome aa3) redox status in turtles (Pseudymys scripta) and rats. Although average brain O2 tension (PtO2) during normoxia was similar in turtle telencephalon and rat parietal cortex, a greater frequency of high and low values of PtO2 occurred in rats, which may indicate a steeper O2 gradient between capillaries and cells. When the fractional inspired O2 concentration (FIO2) was increased, PtO2 was elevated and cytochrome aa3 became more oxidized, whereas decreased FIo2 produced opposite changes in both species. In turtle brain, however, the relationship between PtO2 and cytochrome aa3 redox state was nearly linear over a wide range of PtO2 values. In rats, this relationship was steeper at PtO2 below normoxia and approached a plateau at Pto2 values above normoxia. Turtle brain cytochrome aa3 appeared more reduced in normoxia, which may be due either to lower affinity for O2 or to enhanced substrate supply. These results indicate that differences in resistance to hypoxia/anoxia observed between rats and turtles are probably not due to differences in O2 availability. Rather, it is likely that differences in redox activities of cytochrome aa3 and/or in substrate use play a role in the relative insensitivity of turtle brain to O2 deprivation. PMID- 6295992 TI - Effect of converting enzyme inhibition on pulmonary edema after microembolization. AB - We examined the effect of converting enzyme inhibition with captopril (SQ 14,225) on pulmonary hemodynamics and on accumulation of extravascular lung water after microembolization in dogs. Pulmonary microembolization, induced with glass beads (200 microns diam), increased the mean pulmonary arterial pressure to approximately 40 Torr in both control and captopril-treated animals. The increase in pulmonary vascular resistance (PVR) in control animals was sustained during the 75 min of study and was associated with an increase in the extravascular lung water content-to-bloodless dry lung weight ratio (W/D) from a normal value of 2.84 +/- 0.22 to 4.53 +/- 0.24 ml/g (P less than 0.01) after embolization. In the captopril-treated group, the PVR increased gradually, such that the value at 75 min postembolization was greater than in controls (P less than 0.05). The W/D in the captopril-treated group of 4.62 +/- 0.19 ml/g was greater than the value of 2.83 +/- 0.10 ml/g in nonembolized captopril-treated animals, but the degrees of edema in the control and captopril-treated animals were not different. A similar degree of embolization induced during infusion of 5 micrograms X kg-1 X h-1 of bradykinin also did not enhance the pulmonary edema, and there was also a greater increase in PVR than in control animals after embolization. These findings suggest that bradykinin does not contribute to the degree of pulmonary edema after microembolization. PMID- 6295993 TI - Ceftizoxime and other third-generation cephalosporins: structure-activity relationships. PMID- 6295994 TI - Ceftizoxime, a broad-spectrum beta-lactamase stable cephalosporin. PMID- 6295995 TI - Comparative tissue and extravascular fluid concentrations of ceftizoxime. PMID- 6295996 TI - Tissue penetration characteristics of ceftizoxime and cefazolin in human bile and gallbladder wall. PMID- 6295997 TI - Human cerebrospinal fluid pharmacokinetics and treatment of bacterial meningitis with ceftizoxime. PMID- 6295998 TI - Lack of disulfiram--like reactions with ceftizoxime. PMID- 6295999 TI - The use of ceftizoxime in the treatment of critically ill patients infected with multiply antibiotic resistant bacteria. PMID- 6296000 TI - Ceftizoxime in moderate-to-severe infections. PMID- 6296001 TI - Ceftizoxime treatment of infection in neutropenic patients with malignancies. PMID- 6296002 TI - Ceftizoxime in the treatment of infections in patients with cancer. PMID- 6296003 TI - The efficacy of ceftizoxime in the therapy of bacteraemia. PMID- 6296004 TI - Evaluation of ceftizoxime in acute peritonitis. PMID- 6296005 TI - A preliminary report on the use of ceftizoxime vs. clindamycin/tobramycin for the therapy of intra-abdominal and pelvic infections. PMID- 6296006 TI - The efficacy of ceftizoxime in treating infections due to organisms resistant to other antibiotics. PMID- 6296007 TI - Ceftizoxime treatment of pneumonia, cellulitis and other infections in 120 hospitalized patients. PMID- 6296008 TI - A randomized clinical trial of ceftizoxime and cefamandole in the treatment of serious lower respiratory tract infections. PMID- 6296009 TI - Comparative study of ceftizoxime and cefamandole in the treatment of bronchopulmonary infections. PMID- 6296011 TI - Ceftizoxime and cefamandole in adult patients with acute respiratory failure. PMID- 6296010 TI - Single-blind controlled study of ceftizoxime and cefamandole in the treatment of community-acquired pneumonia. PMID- 6296012 TI - Comparison of ceftizoxime and penicillin for the treatment of uncomplicated gonorrhoea. PMID- 6296013 TI - Ceftizoxime for treatment of gonorrhoea. PMID- 6296014 TI - A double-blind evaluation of ceftizoxime versus cefamandole therapy for urinary tract infections. PMID- 6296016 TI - Ceftizoxime: collaborative multiphased in-vitro evaluation including tentative interpretive standards for disc susceptibility tests, beta-lactamase stability, and inhibition. PMID- 6296015 TI - Ceftizoxime in the treatment of urinary tract infection in spinal cord injury patients: comparison with tobramycin. PMID- 6296017 TI - Ceftizoxime therapy of infections in hospitalized patients and comparison with cefamandole for urinary tract infections. PMID- 6296018 TI - A comparative trial of ceftizoxime versus cefamandole in the treatment of acute urinary tract infections. PMID- 6296019 TI - Treatment of acute and chronic osteomyelitis with ceftizoxime. PMID- 6296020 TI - Ceftizoxime compared with cefamandole for treatment of soft tissue infections. PMID- 6296021 TI - Comparative study of cefazolin, cefoxitin, and ceftizoxime for surgical prophylaxis in colo-rectal surgery. PMID- 6296022 TI - A randomized controlled study comparing ceftizoxime, cefamandole, and cefoxitin in obstetric and gynaecological surgery: a preliminary report. PMID- 6296023 TI - Use of ceftizoxime in the treatment of paediatric infections. PMID- 6296024 TI - The use of ceftizoxime in neonates. PMID- 6296025 TI - Summary of comparative clinical studies of ceftizoxime and cefamandole, cefazolin and tobramycin. PMID- 6296027 TI - Ceftizoxime: clinical evaluation of efficacy and safety in the U.S.A. PMID- 6296026 TI - Experience with ceftizoxime: an overall summary of clinical trials in Japan. PMID- 6296028 TI - In-vitro activity of ceftizoxime against anaerobic bacteria and comparison with other cephalosporins. PMID- 6296029 TI - The activity of ceftizoxime in combination with gentamicin against hospital isolates of gentamicin-susceptible and resistant Gram-negative bacteria. PMID- 6296030 TI - Synergism of ceftizoxime and tobramycin for Pseudomonas aeruginosa. PMID- 6296031 TI - Combination of ceftizoxime with azlocillin, mezlocillin, piperacillin and ticarcillin. PMID- 6296032 TI - Synergism and antagonism of ceftizoxime and other new cephalosporins. PMID- 6296033 TI - Efficacy of ceftizoxime and related compounds in animals models of infection. PMID- 6296034 TI - Pharmacokinetics of ceftizoxime. PMID- 6296036 TI - The effect of dexamethasone on the replication of herpes simplex virus in human gingival fibroblast cultures. PMID- 6296035 TI - Comparative pharmacokinetics of ceftizoxime and other third-generation cephalosporins in humans. PMID- 6296038 TI - Evidence for a plasmid in a methanogenic bacterium. AB - Among 15 strains of methanogens, one plasmid, pMP1, was identified in the new coccoid isolate PL-12/M. It could not be detected in the cleared lysate, but it was detected in the viscous pellet. The plasmid had a molecular weight of ca. 4.6 x 10(6). A restriction enzyme cleavage map of the cloned plasmid was derived. PMID- 6296037 TI - Ordering of the flagellar genes in Pseudomonas aeruginosa by insertions of mercury transposon Tn501. AB - The flagellar genes of Pseudomonas aeruginosa PAO cluster on the chromosome at two distinct regions, region I and region II. The order of the flagellar cistrons in this organism was established by using transducing phage G101 and plasmids FP5 and R68.45. A method to insert transposon Tn501 near the fla genes was devised. We obtained two strains in which Tn501 was inserted at sites close to the flagellar cistrons in region II. We isolated Fla mutants in which the chromosomal segment between the two Tn501 insertion sites was deleted. Using Tn501-encoded mercury resistance as an outside marker, we determined the order of 9 of the 11 flagellar cistrons in region II as follows: puuF-region I-flaG-flaC-flaI-flaH flaD-flaB-flaA-flaF-flaE-pur-67. By using phage G101-mediated transduction, the mutation converting monoflagellated bacteria into the multiflagellated (mfl) form was closely linked to the five fla cistrons in region I. Using mfl as an outside marker, we determined the order of the five cistrons as follows: puuF-flaV-flaZ flaW-flaX-flaY-region II. The mfl mutation was shown to be either located within the flaV cistron or linked very closely to this cistron. No linkage was observed in transductions between any of the fla cistrons in region I and any of the fla cistrons in region II. PMID- 6296039 TI - Genetic mapping of the minB locus in Escherichia coli K-12. AB - The minB (minicell production) locus of Escherichia coli K-12 was mapped by transduction using bacteriophage P1. minB is located at min 25.6, between purB (min 25.2) and dadR (min 25.8). The mapping was facilitated by the use of insertion zcf-236::Tn10, which is inserted at min 25.4. PMID- 6296040 TI - Role of the sfiA-dependent cell division regulation system in Escherichia coli. AB - Several authors have suggested that the SOS-associated (sfiA-dependent) system of division inhibition, normally induced by perturbations of DNA replication, also regulates steady-state (unperturbed) cell division. The present work shows that mean cell mass is identical in sfiA+ and sfiA mutant cultures during steady-state growth, that mass adjustment is identical after shift up, that sfiA expression is not induced by shift up, and that a sfiA mutation does not cause aberrant chromosome segregation. PMID- 6296042 TI - Location on the Salmonella typhimurium chromosome of the gene encoding nucleoside diphosphokinase (ndk). AB - The gene encoding nucleosidediphosphate kinase (ndk) was located at 55 units on the Salmonella typhimurium chromosome. The ndk locus was 83% cotransducible with hisS and 2% cotransducible with glyA in phage P22-mediated crosses. A nucleosidediphosphate kinase mutant that produced only 10% of the wild-type enzyme activity (ndk-1) grew normally and produced a heat-labile enzyme. PMID- 6296041 TI - In vivo packaging of cosmids in transposon-mediated mutagenesis. AB - A technique was developed that permits the analysis of large regions of DNA by transposition mutagenesis. Large fragments of the pTiA6NC plasmid were cloned into the broad host range cosmid pHK17 and subjected to transposition mutagenesis by Tn3. Cosmids containing Tn3 insertions were selected by in vivo packaging by lambda cI857 and transduction to a new host. The insertions were localized by DNA restriction endonuclease analysis and transferred to the Ti-plasmid by marker exchange. PMID- 6296043 TI - Cloning of the aerobactin-mediated iron assimilation system of plasmid ColV. AB - The high-affinity iron assimilation system of plasmid ColV-K30 was cloned on the vector plasmid pPlac. Plasmid pABN1 was isolated by means of sensitivity to cloacin, a bacteriocin using the same outer membrane receptor as ferric aerobactin. Restriction maps were determined for this plasmid and for a subclone, pABN5. Plasmid pABN1 codes for the complete gene complex, whereas plasmid pABN5 encodes only the biosynthetic genes for aerobactin. Regulation of the uptake system by iron is retained in cloned sequences of pABN1. PMID- 6296044 TI - Internal promoters of the his operon in Salmonella typhimurium. AB - Two internal promoters in the his operon of Salmonella typhimurium have been precisely mapped genetically. The internal promoters are found in, or very close to, gene border regions in the his operon. The his operon was examined for the presence of additional internal promoters whose transcripts were sensitive to rho mediated transcription termination and therefore had escaped detection. No new internal promoters were found. It is argued that the internal promoters described here are not likely to be fortuitous message start sites, but may play a physiologically important role in operon expression. PMID- 6296045 TI - Cloning, mapping, and expression of the fumarase gene of Escherichia coli K-12. AB - Two classes of fumarase-transducing phages, lambda fumA and lambda fumB, were isolated from populations of recombinant phages containing HindIII fragments of Escherichia coli DNA; they were isolated by virtue of their ability to complement the metabolic lesion of a fumarase-negative mutant. The strongly complementing lambda fumA phages contained a 6.2-kilobase HindIII fragment encoding: the fumA gene, located at 35.5 min in the E. coli linkage map and expressing the major fumarase activity; the mannosephosphate isomerase gene, manA; and an unidentified gene, g48. The three genes were located relative to the restriction map of the cloned fragment and the genetic linkage map (terC-g48-fumA-manA-uidAoR), their transcription polarities were defined as anticlockwise in the chromosome, and the molecular weights of the corresponding gene products were established: fumA, 61,500; manA, 42,000; g48, 48,000. Organisms containing the fumA gene sub-cloned in multicopy plasmids overproduced fumarase up to 50-fold. The weakly complementing class of transducing phages, lambda fumB, contained several genes in an 8.2-kilobase HindIII fragment, including one (fumB) that determines a minor fumarase activity. Complementation by fumB was only observed in high-copy situations such as transduction plaques and in strains containing a multicopy plasmid in which 40% of normal fumarase activity was detected. The basis for the complementation by fumB was not defined. PMID- 6296046 TI - Molecular cloning of the region of the terminus of Escherichia coli K-12 DNA replication. AB - A series of plasmids have been isolated either by ligation of defined restriction fragments to plasmid pBR325 or by screening of a cosmid bank by in situ colony hybridization. Together with one previously isolated plasmid, they spanned 86% of the 30.5- to 34-min region of the genetic map of Escherichia coli K-12. Physical analysis of these plasmids and hybridizations to Southern blots confirmed the endonuclease map of this region, with the exception of a 9.3-kilobase pair inversion. PMID- 6296047 TI - Origin of Escherichia coli K-12 Hfr B7. AB - Several F' plasmids encoding resistance to tetracycline have been derived from a trg::Tn10 Hfr B7 strain of Escherichia coli K-12. One of these plasmids, JGF312, was analyzed by restriction endonuclease digestion and Southern blot hybridization to cloned chromosomal fragments. This analysis revealed that JGF312 was formed by Tn10-promoted deletion from the Tn10 insertion (31.4 min) to within the prophage rac at 30.1 min. Hfr B7 was shown to result from recombination between IS2 of F delta (33-43) and a chromosomal IS2 located within the rac-man region at 30.9 min on the genetic map. PMID- 6296048 TI - Localization of symbiotic mutations in Rhizobium meliloti. AB - A total of 5 Nod- and 57 Fix- symbiotic mutants of Rhizobium meliloti strain 41 have been isolated after either nitrosoguanidine or Tn5 transposition mutagenesis. Chromosomal locations of mutations in 1 Nod- and 11 Fix- derivatives were ascertained by transferring the chromosome (mobilized by plasmid R68.45), in eight fragments, into symbiotically effective recipients and testing the recombinants for symbiotic phenotype. Alternatively, the kanamycin resistance marker of Tn5 was mapped. In five mutants the fix alleles were localized on different chromosomal regions, but six other fix mutations and one nod mutation tested did not map onto the chromosome. It was shown that the chromosome mobilizing ability (Cma+) of R68.45 was not involved in the mobilization of genes located extrachromosomally. Moreover, Cma- derivatives of R68.45 could mobilize regions of the indigenous plasmid pRme41b but not chromosomal genes. Thus, mobilization of a marker by Cma- R68.45 indicates its extrachromosomal location. With a 32P-labeled DNA fragment carrying Tn5 as a hybridization probe, it was shown that in five extrachromosomally located Tn5-induced fix mutants and one nod mutant Tn5 was localized on plasmid pRme41b. This is in agreement with the genetic mapping data. PMID- 6296049 TI - Regulation of yeast trehalase by a monocyclic, cyclic AMP-dependent phosphorylation-dephosphorylation cascade system. AB - Mutation at the GLC1 locus in Saccharomyces cerevisiae resulted in simultaneous deficiencies in glycogen and trehalose accumulation. Extracts of yeast cells containing the glc1 mutation exhibited an abnormally high trehalase activity. This elevated activity was associated with a defective cyclic AMP (cAMP) dependent monocyclic cascade which, in normal cells, regulates trehalase activity by means of protein phosphorylation and dephosphorylation. Trehalase in extracts of normal cells was largely in a cryptic form which could be activated in vitro by ATP . Mg in the presence of cAMP. Normal extracts also exhibited a correlated cAMP-dependent protein kinase which catalyzed incorporation of label from [gamma 32P]ATP into protamine. In contrast, cAMP had little or no additional activating effect on trehalase or on protamine phosphorylation in extracts of glc1 cells. Similar, unregulated activation of cryptic trehalase was also found in glycogen deficient strains bearing a second, independently isolated mutant allele, glc1-2. Since trehalase activity was not directly affected by cAMP, the results indicate that the glc1 mutation results in an abnormally active protein kinase which has lost its normal dependence on cAMP. Trehalase in extracts of either normal or mutant cells underwent conversion to a cryptic form in an Mg2+-dependent, fluoride-sensitive reaction. Rates of this reversible reduction of activity were similar in extracts of mutant and normal cells. This same, unregulated protein kinase would act on glycogen synthase, maintaining it in the phosphorylated low activity D-form. The glc1 mutants provide a novel model system for investigating the in vivo metabolic functions of a specific, cAMP-dependent protein kinase. PMID- 6296050 TI - Multiple genes for membrane-bound phosphatases in Escherichia coli and their action on phospholipid precursors. AB - We have devised a coupled radiochemical assay for detecting phosphatidylglycerolphosphate (PGP) phosphatase activity in Escherichia coli colonies immobilized on filter paper. There appeared to be at least two enzymes capable of dephosphorylating PGP, as judged by the characterization of mutations in two genes designated pgpA and pgpB. The former is located near min 10 and is cotransducible with proC and dnaZ. The latter is situated near min 28 and is closely linked to cysB. The available mutant alleles of pgpA reduced the specific activity of PGP phosphatase in crude extracts by about 30%, but they had no effect on phosphatidic acid (or lysophosphatidic acid) phosphatase. Mutants altered in the pgpB locus inactivated most of the residual PGP phosphatase activity present in single-step pgpA mutants, and the level of phosphatidic acid phosphatase was also reduced 20-fold. The available mutations in pgpA and pgpB elevated the cellular PGP pool by 10- to 50-fold. The maximal PGP levels never exceeded 5%, and these strains were not conditionally lethal. The simplest interpretation of our findings is that there are at least two membrane-associated phosphatases in E. coli, both distinct from alkaline phosphatase. The pgpA gene product is specific for PGP, whereas the pgpB gene product also acts on phosphatidic acid and lysophosphatidic acid. PMID- 6296051 TI - pH-sensitive CDP-diglyceride synthetase mutants of Escherichia coli: phenotypic suppression by mutations at a second site. AB - In Escherichia coli, mutations which lower the level of CDP-diglyceride synthetase are designated cds and map at min 4. The cds-8 mutation resulted in strikingly defective enzyme activity and also rendered cells pH sensitive for growth. Both the inhibition of growth and the massive accumulation of phosphatidic acid which occur in a cds-8 mutant at pH 8 were suppressed by mutations at a second locus, designated cdsS, which mapped between argG and gltB near min 68. The cdsS3 mutation by itself did not affect CDP-diglyceride synthetase activity in wild-type cells, but it caused a twofold stimulation of the residual activity present in strains harboring cds-8. Both the insensitivity to pH and the twofold stimulation of residual activity were lost by introduction of an F' strain carrying cdsS+ into a recA1 cds-8 cdsS3 host. When a culture of a cds-8 cdsS+ strain was shifted to pH 8, the residual specific activity of synthetase dropped by 75% within 100 min. In a cds-8 cdsS3 double mutant under the same conditions, the activity declined appreciably less, about to the level found in the cds-8 cdsS+ strain under permissive conditions (pH 6). Thus, it appears that mutations in the cdsS gene suppress the pH sensitivity of cds mutants by inhibiting the decay of residual CDP-diglyceride synthetase activity at the nonpermissive pH. The cdsS locus appears to be distinct from any known nonsense or missense suppressor. PMID- 6296053 TI - Isolation and characterization of P1 minireplicons, lambda-P1:5R and lambda P1:5L. AB - We have isolated two new classes of P1 miniplasmids, called lambda-P1:5R and lambda-P1:5L, by the in vivo extension of a cloned P1 fragment, EcoRI-5, which by itself is not capable of plasmid replication. The lambda-P1:5R plasmids contain EcoRI-5 plus a variable portion of the adjacent P1 EcoRI fragment 8. They have a copy number like that of P1 (about 1 per host chromosome), are faithfully segregated at cell division, and are subject to incompatibility exerted by either a single copy of P1 or a single copy of EcoRI-5. In contrast, the lambda-P1:5L plasmids contain EcoRI-5 and a portion of adjacent P1 DNA that includes at least P1 EcoRI fragments 15, 18, and 23 and a part of fragment 17. These plasmids have a copy number of about 15 per cell chromosome. Despite this they are segregated to daughter cells somewhat less faithfully than are lambda-P1:5R plasmids. They are sensitive to incompatibility exerted by a single copy of P1, but not to incompatibility exerted by a single copy of EcoRI-5. lambda-P1:5L plasmids are, however, sensitive to incompatibility exerted by multiple copies of EcoRI-5. These results show that the relative copy numbers of exerting and responding elements are important for the incompatibility phenotype and strongly suggest that lambda-P1:5L plasmids lack a repressor of replication that can be supplied in trans from P1 but not from EcoRI fragment 5. We suggest that P1 normally uses the 5R replicon and that the 5L replicon may be a backup system that ensures plasmid maintenance should the primary replication event fail to initiate. PMID- 6296052 TI - Construction of hybrid plasmids containing the Escherichia coli uxaB gene: analysis of its regulation and direction of transcription. AB - The uxaB gene of Escherichia coli, encoding for altronate oxidoreductase involved in the hexuronate degradative pathway, was isolated on a ColE1-uxaB hybrid plasmid from the Clarke and Carbon bank. The restriction map of this plasmid was established. The uxaB gene was mapped on a 1.5-megadalton HindIII-KpnI DNA fragment. Use of an in vitro gene fusion between uxaB and lacZ genes led to the determination that uxaB is transcribed from the KpnI towards the HindIII restriction sites. Gene amplification in cells containing various uxaB hybrid plasmids allowed us to show a gradation in the level of repression of exu operator sites by the exuR regulatory gene product. PMID- 6296054 TI - Cloning and expression in Escherichia coli of the naphthalene degradation genes from plasmid NAH7. AB - The genes encoding the enzymes responsible for conversion of naphthalene to 2 hydroxymuconic acid (nahA through nahI) are contained on a 25-kilobase EcoRI fragment of an 85-kilobase NAH plasmid of Pseudomonas putida. These genes were cloned into the plasmid vectors pBR322 and RSF1010 to obtain the recombinant plasmids pKGX505 and pKGX511, respectively. To facilitate cloning and analysis, an NAH7 plasmid containing a Tn5 transposon in the salicylate hydroxylase gene (nahG) was used to derive the EcoRI fragment. The genes for naphthalene degradation were expressed at a low level in Escherichia coli strains containing the fragment on the recombinant plasmids pKGX505 or pKGX511. This was shown by the ability of whole cells to convert naphthalene to salicylic acid and by in vitro enzyme assays. The expression of at least two of these genes in E. coli appeared to be regulated by the presence of the inducer salicylic acid. In addition, high-level expression and induction appear to be mediated by an NAH plasmid promoter and a regulatory gene located on the fragment. A restriction endonuclease cleavage map of the cloned fragment was generated, and the map positions of several nah genes were determined by analysis of various subcloned DNA fragments. PMID- 6296055 TI - Genetic map of the opp (Oligopeptide permease) locus of Salmonella typhimurium. AB - The uptake of peptides by Salmonella typhimurium is mediated by three apparently independent transport systems. One of these systems, the oligopeptide permease, is encoded by a genetic locus (opp) which has been mapped at 34 min on the S. typhimurium chromosomal map. We accurately mapped the location of opp by cotransduction frequencies and by deletion analysis and show that the gene order for this region of the chromosome is cysB-trp-tonB-opp-galU-tdk. All opp mutants, independently isolated by a variety of means, mapped at this one locus, between tonB and galU. Spontaneous and transposon Tn10-generated deletions were used to construct a fine-structure genetic map of opp. Evidence is presented which indicates that opp covers a 5- to 6-kb segment of DNA and is therefore likely to consist of more than one gene. PMID- 6296056 TI - Relationship between plasmid and chromosomal hemolysin determinants of Escherichia coli. AB - Plasmid hemolysin (hly) determinants have been shown previously to comprise three cistrons (hlyA, hlyB, hlyC), coding for the synthesis and transport of hemolysin. Using recombinant plasmids as specific probes for these cistrons, we were able to analyze the chromosomal hly determinants of nine Escherichia coli strains which belonged to serotypes O4, O6, O18, and O75 and were isolated from urinary tract infections and fecal flora. The chromosomal hly genes shared extensive sequence homology with the cloned plasmid hly determinant. Nevertheless, small differences were observed, and these were found to lie mainly within cistron A (hlyA), which has been shown to determine the hemolysin protein itself. These fine variations were not specific for the O-serotype. PMID- 6296057 TI - Identification and control of synthesis of the dsdC activator protein. AB - Operon fusions between the D-serine deaminase regulatory and structural genes and lacZ were constructed and used to examine the control of expression of the positive regulatory gene, dsdC. Merodiploid strains containing both dsdCp::Mu d (lac Apr) and dsdC+A+ produced only one-fourth as much beta-galactosidase as did the haploid dsdCp::Mu d (lac Apr) strains, indicating that the dsdC+ product repressed its own synthesis. The repression was reversed by D-serine. dsdC expression was not depressed in a cya background. The basal level of D-serine deaminase was the same in wild-type and dsdCp fusion strains. The dsdC gene product was identified in maxicell strains harboring dsd plasmids as a 34,000 dalton protein. dsdC gene transcription proceeded clockwise; thus, its promoter is adjacent to that of dsdA. PMID- 6296058 TI - Mutational analysis of the virulence region of an Agrobacterium tumefaciens Ti plasmid. AB - Forty-nine Tn3 and Tn5 transposition insertion mutations were introduced into the virulence region of the pTiA6NC plasmid of Agrobacterium tumefaciens. Five Tn5 transposition mutations from an earlier study (D. Garfinkel and E. Nester, J. Bacteriol. 144:732-743, 1980) were also mapped more accurately. These mutations defined five separate loci within the virulence region. Two Tn3 insertions into one of these loci, virA, result in a strain which is only weakly virulent; however, a Tn5 insertion into this locus eliminates virulence. One Tn5 insertion into another locus, virC, results in a strain which is weakly virulent. Two additional Tn5 insertions into this locus eliminate virulence. Insertions into the remaining three loci eliminate virulence entirely. PMID- 6296059 TI - Physical and genetic characterization of the cloned sbcB (exonuclease I) region of the Escherichia coli genome. AB - A 17-kilobase (kb) HindIII fragment containing the structural gene for exonuclease I (sbcB) from Escherichia coli K-12 was physically and genetically characterized. The monomeric molecular weight of exonuclease I was 53,700, based on sodium dodecyl sulfate-polyacrylamide gel electrophoresis of 35S-labeled E. coli mini- and maxicells. The gene was in close proximity to two unidentified proteins with molecular weights of 15,200 and 13,100. No other polypeptides appeared to be constitutively synthesized from the 17-kb fragment. Genetically, no portion of the histidine operon or the shikimic acid transport gene (shiA) was detected on the fragment. Although the entire 17-kb fragment in the vector pMB9 was too unstable to be useful, a 7.6-kb BamHI-EcoRI fragment inserted into a variety of vectors was stable. A detailed restriction map of the fragment is presented. Several derivatives in the runaway-replication vectors pMB06 and pMOB45 yielded 20- to 52-fold increases in exonuclease I activity after a switch in growth temperature to 40 degrees C. Of six exonuclease I mutants examined by DNA-DNA hybridization, one (xonA6) appeared to have arisen from a 1.2-kb insertion into the structural gene for exonuclease I. PMID- 6296060 TI - Analysis of tetracycline resistance encoded by transposon Tn10: deletion mapping of tetracycline-sensitive point mutations and identification of two structural genes. AB - Deletions in the tet genes derived from Tn10 were formed from different tet::Tn5 insertion mutations by removing DNA sequences located between a HindIII site in Tn5 and a HindIII site adjacent to the tet genes. Tetracycline-sensitive point mutations were mapped in recombination tests with the deletions and were thus aligned with the genetic and physical map of the tet region. Plasmids carrying point mutations were tested for complementation with derivatives of pDU938, a plasmid carrying cloned tet genes derived from Tn10 which had been inactivated by Tn5 insertions. Complementation occurred between promoter-proximal tet point mutations and distal tet::Tn5 insertions, suggesting the existence of two structural genes, tetA and tetB. These results, together with the analysis of polypeptides in minicells harboring pDU938tet::Tn5 mutants, suggested that tetA and tetB are expressed coordinately in an operon. The tetB gene encodes the previously characterized 36,000-dalton cytoplasmic membrane TET protein, but the product of tetA was not identified. Point mutations in either tetA or tetB led to the defective expression of the resistance mechanism involving tetracycline efflux. It is suggested that the tetA and tetB products interact cooperatively in the membrane to express resistance. PMID- 6296062 TI - Properties of a DNA-binding protein in a culture medium of thymus cells. AB - A DNA-binding protein, which migrated as one major protein band, with a molecular weight of 14,000, on sodium dodecylsulfate polyacrylamide gel, was purified from a culture medium of mouse thymus cells. The interaction of the isolated protein with DNA in vitro was assayed by a nitrocellulose filter binding technique. Equilibrium competition experiments demonstrated that the DNA-binding protein had the ability to differentiate among sequences of polynucleotides, indicating that the DNA-binding protein-DNA interaction was at least partially specific. This protein increased the helix melting temperature of DNA and inhibited the incorporation of [3H]dTMP into DNA by the DNA polymerase of calf thymus in vitro. PMID- 6296061 TI - p lambda CM system: observations on the roles of transposable elements in formation and breakdown of plasmids derived from bacteriophage lambda replicons. AB - Transduction with phage derived from a 2-year-old lysate of lambda cam105 (lambda::Tn9) gave rise to chloramphenicol-resistant (Cm(r)) transductants harboring a plasmid (plambdaCM1) formed from lambda cam105 by a Tn9-mediated adjacent deletion to position 36.07 kilobases in the N cistron of lambda. The plambdaCM element can replicate as a plasmid, insert into the bacterial genome, or reproduce lytically as a phage on cells that provide N function. The feasibility of obtaining high titers in encapsidated form and the ease of synchronous introduction into and recovery from bacterial populations make plambdaCM very suitable for quantitative studies of recombination involving transposable elements. Replicon fusions between plambdaCM1 and RSF1596 (pMB8::Tn3Delta596) occur by duplication of either IS1 (at low rate in the absence of TnpA activity) or Tn3Delta596 (in the presence of TnpA activity). At 24 or 32 degrees C, the rate of increase of TnpA-mediated fusions per plambdaCM is about 2% per cell doubling. RSF103 contains the deleted Tn1DeltaAp (which lacks intact beta-lactamase and TnpR resolvase coding sequences) adjacent to a streptomycin resistance (Sm(r)) determinant. We observed that Tn1DeltaAp mediates insertions of external RSF103 sequences into the R388 plasmid. R388::Tn1DeltaAp plasmids show transposition immunity in cells lacking TnpR activity. Using the plambdaCM system, we isolated adjacent transpositions of the RSF103 Sm(r) determinant. The resulting plambdaCM-Sm cosmids contain Sm(r) genetic material flanked by direct repeats of Tn1DeltaAp, and all are deleted for some RSF103 or plambdaCM sequences. The plambdaCM-Sm constructs will fuse into R388 by duplication of a single Tn1DeltaAp element. In the presence of tnpR(+) (but not tnpR) Tn1 or Tn3 elements, all Tn1DeltaAp-mediated complex replicons break down completely and rapidly to simple Tn1DeltaAp inserts. The equilibrium for resolution is at least 10(5):1, and resolution is more than 90% complete after 40 min of exposure to a tnpR(+) cytoplasm. In the absence of TnpR, Rec, and Red activities, Tn1DeltaAp-mediated complex replicons yield simple Tn1DeltaAp inserts at a lower rate. The presence of intact RSF103 replication determinants between direct Tn1DeltaAp repeats appears to accelerate this precise TnpR- and Rec independent breakdown. PMID- 6296064 TI - Orientation of platelets on the surfaces of glass and Teflon plates as studied by a spin label technique. AB - Platelets were found by a spin label technique to orient on flat surfaces of glass as well as of Teflon. This kind of platelet orientation was not caused by centrifugation or partial dehydration of the membrane preparation as employed usually to make oriented planar multilayers of biological membranes on the surface of a supporting plate (1-8), but was considered to be closely related to the adhesion or the aggregation properties of platelets. The amount of oriented platelets varied depending on the platelet treatment and was estimated from a computer simulation of the observed ESR spectra to be about one-half of that of the non-oriented ones in the case of thrombin-treated platelets. This technique may be useful as a new tool to explore the adhesion or aggregation properties of platelets. PMID- 6296063 TI - Membrane-bound cytochromes c of Pseudomonas aeruginosa grown aerobically. Purification and characterization of cytochromes c-551 and c-555. AB - Membrane-bound cytochromes c of Pseudomonas aeruginosa grown aerobically were investigated. By detecting polypeptides with heme-catalyzing peroxidase activity on a sodium dodecyl sulfate polyacrylamide gel, four major (Band I, 33,000 daltons; II, 25,000; III, 20,000; IV, 16,000) and one minor (V, 11,500) hemoproteins were found in the membrane fraction, while one hemoprotein (VI, 8,200) was detected in a small amount in the cytosol fraction. All these hemoproteins (bands I to VI) appeared to be cytochromes c, because all bands were detected even after being treated with HCl-acetone. Of the membrane-bound cytochromes c, cytochromes c-551 (band IV) and c-555 (band V) were solubilized with Triton X-100 and purified by repeated DEAE-cellulose column chromatography. Both purified cytochromes c-551 and c-555 were monomeric and their molecular weights were estimated to be 16,400 and 11,500, respectively. Their respective midpoint potentials were 0.31 and 0.34 V, and their respective isoelectric points in the reduced form were 3.8 and 5.2. The purified cytochromes c-551 and c-555 were found to be clearly different from "soluble" cytochrome c-551, and might function in the membrane-bound aerobic respiratory chain of P. aeruginosa. PMID- 6296065 TI - Substrate specificity and mode of action of the zinc-metallo nuclease from Physarum polycephalum. AB - The alkaline zinc-metallo nuclease of Physarum polycephalum is an endonuclease with a high specificity for single-stranded nucleic acids. Single-stranded DNA was cleaved at least 6,000 times faster than double-stranded DNA under identical conditions. In the supercoil-induced single-stranded region of Form I PM2 DNA only a single nick was made. The nuclease showed nucleotide specificity. Poly(A), poly(I), and poly(dT) were preferentially hydrolyzed. Product analysis showed that it acted by an endonucleolytic mechanism: long polynucleotides were fragmented via intermediate length products to oligo- and mono-nucleotides with the phosphate group at the 5'-terminal position. Extensive similarities exist with the single-strand-specific nuclease S1 from Aspergillus. The zinc-metallo endonuclease from Physarum could be used as a similar probe for single-stranded nucleic acids at neutral or alkaline pH conditions. PMID- 6296066 TI - Probable involvement of both intrachain cAMP binding sites in activation of protein kinase. PMID- 6296067 TI - Evidence that cyclic nucleotides activating rabbit muscle protein kinase I interact with both types of cAMP binding sites associated with the enzyme. AB - Eighty different adenine-modified cAMP analogs were tested as activators of rabbit muscle protein kinase I (cAKI) in an in vitro phosphotransferase assay. All the analogs tested were able to activate completely the kinase. The affinities of the cAMP derivatives for the two types (A and B) of binding sites associated with the regulatory moiety of cAKI were determined under conditions similar to those used in the phosphotransferase assay. The potency of the cAMP analogs as cAKI activators was found to correlate with the mean affinity for sites A and B, rather than to the affinity for only one of the sites. This was true whether cAKI was assayed at low or near physiological ionic strength, whether the concentration of cAKI binding sites was 0.2 or 400 nM, and whether the kinase substrate was mixed histones or homogeneous phenylalanine-4 monooxygenase. Furthermore, site A-selective and site B-selective cAMP analogs activated cAKI synergistically. Finally, it was shown that the degree of synergism between cAMP analogs in activating cAKI correlated with their degree of site selectivity. It is concluded that cyclic nucleotides interact with both types of binding sites in the process of cAKI activation. PMID- 6296068 TI - Temperature and pH dependence of the proton magnetic hyperfine resonances of cytochrome c peroxidase-cyanide. Evidence for hindered vinyl group rotation as a mediator of the enzymes activity. AB - Proton nmr studies of the hyperfine resonances of cytochrome c peroxidase reveal that two pH-dependent processes can be monitored. One of these is the simple pH titration of a resonance which has been previously assigned to the alpha-vinyl proton at heme position 4. Combined with this proton's temperature dependence, the pH data indicate that the rotational position of vinyl 4 is changing with a pK which is similar to that which regulates the enzyme's activity. The second process, slow on the nmr time scale, occurs above pH 8. This is beyond what is normally considered to be the optimum pH range for cytochrome c peroxidase's activity and we interpret this to indicate a protein conformational change. PMID- 6296069 TI - The regulation of ubiquinone-6 biosynthesis by Saccharomyces cerevisiae. AB - Increasing concentrations of glucose (1-5%) in the growth medium depressed ubiquinone-6 biosynthesis in continuously cultured wild type Saccharomyces cerevisiae. In addition, an early intermediate in the pathway of ubiquinone-6 biosynthesis, i.e. 3,4-dihydroxy-5-hexaprenylbenzoate (3,4-DHHB), was found to accumulate. The increase in 3,4-DHHB levels varied inversely with the diminished levels of ubiquinone-6, suggesting that O-methylation of 3,4-DHHB is a regulated step in catabolite repression. Experiments using protoplasts demonstrated that the effect of catabolite repression on this pathway was reversible by 1.2 mM cAMP but not by other nucleotides and cyclic nucleotides. This response to cAMP was unaltered by the protein synthesis inhibitor cycloheximide, indicating that the regulatory control for this reaction must occur at the enzymatic level. Additional experiments demonstrated the presence of a heat-labile component of the cytoplasm, which was essential for this effect of cAMP. This observation suggests that this cytosolic effector may be translocated to the inner membrane of the mitochondria, the intracellular site for ubiquinone-6 biosynthesis. PMID- 6296070 TI - Characterization of the association of tritiated enkephalin with neuroblastoma cells under conditions optimal for receptor down regulation. AB - The uptake of tritium-labeled [D-Ala2,D-Leu5]enkephalin ([3H]DADLE) by mouse neuroblastoma cells (N4TG1) was investigated under conditions which are optimal for ligand-induced receptor loss (down regulation). Uptake of [3H]DADLE was a receptor-mediated process, since it was inhibited by opiate receptor ligands and the (i) time course, (ii) dose-response curve, and (iii) temperature dependence of uptake were similar to those for enkephalin-receptor down regulation. Cells in suspension showed less uptake than those in monolayer culture and both uptake and down regulation were decreased by the inhibitors of metabolic energy production, sodium azide, and 2,4-dinitrophenol. Comparison of the effects of these metabolic inhibitors on the processes of receptor loss and ligand uptake showed that these cells accumulate [3H]DADLE in excess of their surface receptor number, suggesting that receptor recycling normally occurs under the conditions studied. The lysosomotrophic amines, chloroquine and methylamine, inhibited dissociation of cell-associated [3H]DADLE but did not affect down regulation. The data are consistent with the idea that enkephalin is internalized via receptor-mediated endocytosis. The possible fate of the "down-regulated" receptors is considered. PMID- 6296071 TI - Proteolytic activation of calcium-activated, phospholipid-dependent protein kinase by calcium-dependent neutral protease. AB - A Ca2+-dependent protease I), which hydrolyzes casein at Ca2+ concentrations lower than the 10(-5) M range, is purified roughly 4000-fold from the soluble fraction of rat brain. This protease is able to activate Ca2+-activated, phospholipid-dependent protein kinase (protein kinase C) by limited proteolysis analogously to the previously known Ca2+-dependent analogously to the previously known Ca2+-dependent protease (Ca2+ protease II) which is active at the millimolar range of Ca2+ (Inoue, M., Kishimoto, A., Takai, Y., and Nishizuka, Y. (1977) J. Biol. Chem. 252, 7610-7616). The protein kinase fragment thus produced shows a molecular weight of about 5.1 X 10(4), and is significantly smaller than native protein kinase C (Mr = 7.7 X 10(4). Although protein kinase C may be normally activated in a reversible manner by the simultaneous presence of phospholipid and diacylglycerol at Ca2+ concentrations less than 10(-6) M, this enzyme fragment is fully active without any lipid fractions and independent of Ca2+. The limited proteolysis of protein kinase C is markedly enhanced in the velocity by the addition of phospholipid and diacylglycerol, which are both required for the reversible activation of the enzyme. However, casein hydrolysis by this protease is not affected by phospholipid and diacylglycerol. Available evidence suggests that, at lower concentrations of this divalent cation, Ca2+ protease I reacts preferentially with the active form of protein kinase C which is associated with membrane, and converts it to the permanently active form. In contrast, the inactive form of protein kinase C, which is free of membrane phospholipid, does not appear to be very susceptible to the proteolytic attack. It remains unknown, however, whether this mechanism of irreversible activation of protein kinase C does operate in physiological processes. It is noted that Ca2+ protease II, which is active at higher concentrations of Ca2+, proteolytically activates protein kinase C irrespective of the presence and absence of phospholipid and diacylglycerol. PMID- 6296072 TI - Mechanism of ATP quench of phosphodiesterase activation in rod disc membranes. AB - GTP-dependent light activation of cyclic GMP phosphodiesterase in bovine rod disc membranes was quenched by ATP. ATP reduced both initial velocity (V0) and turn off time (toff) of phosphodiesterase activated by a flash that bleached 1.5 X 10( 5) of the rhodopsin present. In the absence of rhodopsin kinase, ATP had no effect on either V0 or toff of reconstituted preparations containing phosphodiesterase and GTP*-binding protein. Addition of partially purified rhodopsin kinase to such reconstitutions again permitted ATP to quench both initial velocity and turn off time. It is thus likely that kinase-mediated phosphorylation of bleached rhodopsin reduces and arrests light-induced phosphodiesterase activation. Thermolysin cleavage of rhodopsin's COOH-terminal dodecapeptide eliminated ATP's effect on toff, but did not diminish its effect on V0. Thus, the effects of ATP and kinase on V0 may be mediated by sites proximal to and effects on toff by sites distal to the thermolysin cleavage point at rhodopsin's COOH-terminal end. PMID- 6296073 TI - Identification of bacteriophage T4 gene 60 product and a role for this protein in DNA topoisomerase. AB - Bacteriophage T4 DNA topoisomerase has been isolated and shown to contain the proteins coded by the DNA-delay genes 39 and 52 (Liu, L. F., Liu, C.-C., and Alberts, B. M. (1979) Nature (Lond.) 281, 456-461 and Stetler, G. L., King, G. J., and Huang, W. M. (1979) Proc. Natl. Acad. Sci. U. S. A. 76, 3737-3741). From complementation measurements in vitro and from earlier genetic evidence, these workers suggested that the product of gene 60 (p60) was also a component of the DNA topoisomerase complex. This paper now establishes the identity of p60 and unequivocally shows that this protein is a component of the enzyme complex. T4 DNA topoisomerase was purified by a simplified two-column procedure and found to be a stable complex of p39, p52, and a protein with a relative molecular weight of 18,000. The 18,000-dalton chain has been unambiguously shown to be the product of gene 60 through the use of an amber mutant of gene 60 with Sup+ and Sup- hosts and analyses by two-dimensional gel electrophoresis. While p39 and p52 were tightly associated in the wild type enzyme complex, they were readily separated on a hydroxylapatite column from extracts of cells infected by an amber mutant of gene 60. These findings suggest that p60 plays a structural/functional role in the enzyme complex by holding the larger p39 and p52 in juxtaposition. PMID- 6296074 TI - DNA sequence of the gene coding for Escherichia coli ribonuclease H. AB - The gene for Escherichia coli ribonuclease H has been studied by use of a plasmid which contains a segment of the E. coli chromosome. The genomic DNA was subcloned from pLC28-22 to pBR322 by use of various restriction enzymes. Such subcloning limited the RNase H gene to a piece of DNA no longer than 760 base pairs. Cells bearing plasmids containing the RNase H gene produce as much as 10-15 times the normal amount of RNase H without any drastic effect on maintenance of the plasmid or cell growth. DNA sequence analysis has permitted the prediction of a protein whose molecular weight is 17,559 (155 amino acid residues). The predicted sequence was confirmed by amino acid analysis, NH2-terminal amino acid sequence, and size determination of highly purified RNase H. PMID- 6296075 TI - Regulation of cytochrome P-450. Immunochemical quantitation of eight isozymes in liver microsomes of rats treated with polybrominated biphenyl congeners. AB - Specific immunochemical techniques were used to quantitate the levels of eight isozymes of cytochrome P-450 (P-450) and epoxide hydrolase in liver microsomes of untreated rats and rats treated with phenobarbital, 3-methylcholanthrene, a mixture of these two compounds, nine individual polybrominated biphenyl (PBB) congeners, and a commercial mixture of PBBs. Levels of two 3-methylcholanthrene inducible P-450s (designated P-450 beta NF-B and P-450 beta NF/ISF-G) varied over two orders of magnitude and were highly correlated. The levels of four phenobarbital-inducible P-450s (designated P-450PB-B, P-450PB-C, P-450PB-D, and P 450PB/PCN-E) were all correlated to each other. The level of one form, P-450UT-A, which was present at substantial levels in untreated rats, was inversely correlated to the levels of P-450 beta NF-B and P-450 beta NF/ISF-G. Among the PBB congeners which were examined, the presence of bromine at carbons o to the biphenyl bridge favored the induction of P-450PB-B, P-450PB-C, P-450PG-D, and P 450PB/PCN-E but did not necessarily eliminate the ability to induce P-450 beta NF B and P-450 beta NF-ISF-G. PBB congeners with 2,2'-dibromo substitution induced P 450 beta NF-B and P-450 beta NF/ISF-G if one of the biphenyl rings contained bromines at positions 2,3, and 4. The induction of P-450UT-F was found to occur to a small extent with three of the compounds and is not readily explained in terms of structure-activity relationships. Although correlations were found among levels of some of the forms of P-450, several important exceptions were noted in relative levels of the individual enzymes. While the correlative data are useful in predicting induction patterns, all eight forms of P-450 appear to be independently regulated to some extent. PMID- 6296076 TI - Serum and monoclonal antibodies against the chick intestinal receptor for 1,25 dihydroxyvitamin D3. Generation by a preparation enriched in a 64,000-dalton protein. PMID- 6296077 TI - Cloning and characterization of the rat NADPH-cytochrome P-450 oxidoreductase gene. AB - A rat liver DNA genomic library was prepared using the lambda phage cloning vector Charon 28. Recombinant phage were screened with a cDNA clone (pOR-7) containing sequences complementary to mRNA coding for NADPH-cytochrome P-450 oxidoreductase. This cDNA clone contains the poly(A) addition site and 60% of the mRNA sequence (Gonzalez, F. J., and Kasper, C. B. (1982) J. Biol. Chem. 257, 5962 5968). Four positive phage were identifed and plaque-purified, and their DNA was isolated and subjected to restriction endonuclease mapping. All phage DNA inserts, which ranged from 11 to 16 kilobases, contained several overlapping restriction fragments. The clone with the largest insert (lambda OR-2) was found to contain restriction fragments identical with those of rat DNA when both were subjected to Southern blotting with nick-translated pOR-7 DNA; this finding established the presence in lambda OR-2 of the 3' end (poly(A) addition site) of the oxidoreductase gene. When [32P]cDNA synthesized from enriched oxidoreductase poly(A) RNA was utilized as a probe, additional fragments were identified. The fragment most distal to the 3'-specific fragment was assumed to contain the 5' cap site and was subcloned into pBR322 for further analysis. Restriction mapping and Southern blot analysis further localized the 5' end of the gene to an AvaII fragment of 540 base pairs (bp). Hybridization of this fragment with oxidoreductase mRNA-enriched poly(A) RNA resulted in the arrest of translation of oxidoreductase; this confirmed that it contained an exon region of the oxidoreductase gene. S1 nuclease mapping and DNA sequencing identified to within +/- 1 bp the 5' cap site of the gene which corresponds to an A start. DNA sequencing of the 5'p flanking region revealed no "TATA box" in the vicinity of 25 to -30 bp of the cap site. R-loop analysis of lambda OR-2 revealed the presence of a minimum of seven introns in the 6000-bp oxidoreductase gene and eight exons with a total length of approximately 2600 bp. PMID- 6296078 TI - Location of the single gene for elongation factor Tu on the Euglena gracilis chloroplast chromosome. AB - The structural gene for elongation factor Tu (EF-Tu) has been mapped by heterologous hybridization to a 2900 base pair sequence of Euglena gracilis Klebs Strain Z Pringsheim chloroplast DNA within the EcoRI fragment, Eco N. The hybridization probes were obtained from a HhaI restriction fragment containing internal sequences to Escherichia coli EF-Tu, located in the tufA gene locus, and from an EcoRI restriction fragment of chloroplast DNA from the eukaryotic algae Chlamydomonas reinhardii, containing the 3' end of the chloroplast EF-Tu gene. This is the second identified protein gene locus to be mapped on the E. gracilis chloroplast genome, and the first using prokaryotic DNA as a probe. PMID- 6296079 TI - Molecular size of different neurotoxin receptors on the voltage-sensitive Na+ channel. AB - Measurements were made of the molecular sizes of two distinct receptors on the Na+ channel in rat brain synaptosomes that are specific for different neurotoxins. Radiation inactivation of the binding of radiolabeled derivatives of the toxins was consistent with Mr = 260,000 for the tetrodotoxin receptor and Mr = 266,000 for the receptor specific for two scorpion toxins, toxin II from Centruroides suffusus suffusus and toxin gamma from Tityus serrulatus serrulatus. Covalent cross-linking of the latter to its receptor similarly indicated Mr = 270,000. It seems most likely that these two distinct receptors reside on the same molecule. PMID- 6296080 TI - The relationship between incorporation of E-5-(2-Bromovinyl)-2'-deoxyuridine into herpes simplex virus type 1 DNA with virus infectivity and DNA integrity. AB - E-5-(2-Bromovinyl)-2'-deoxyuridine (BrvdUrd) produced a dose-dependent shift in the density of herpes simplex virus type 1 (HSV-1) DNA at concentrations which yielded potent inhibition of virus replication in cultured Vero cells. Although the density of cellular DNA was not altered by these concentrations of BrvdUrd, incorporation of this analogue into cellular DNA of HSV-1-infected cells has been previously observed in this laboratory. The degree of inhibition correlated with the amount of BrvdUrd substituted for thymidine in HSV-1 DNA. BrvdUrd-substituted DNA was more labile as determined by a dose-dependent increase in single strand breaks when examined by centrifugation in alkaline sucrose gradients. Thus, the potent antiviral action of BrvdUrd observed in cell culture correlates not only with its incorporation into HSV-1 DNA but also with an altered stability of this DNA. PMID- 6296081 TI - Enzymatic activation of hydrazine derivatives. A spin-trapping study. AB - The oxidative metabolism of hydralazine, isoniazid, iproniazid, and phenylhydrazine has been studied using spin-trapping techniques. The oxidation of these hydrazine derivatives, catalyzed by horseradish peroxidase and prostaglandin synthetase, produces reactive free radical intermediates. Enzymatic activation of hydralazine produce the nitrogen-centered hydralazyl radical (RNHNH); phenylhydrazine formed only the phenyl radical. Iproniazid, on the other hand, formed both the isopropyl radical and a hydroperoxy radical. The formation of the hydroperoxy radical was not inhibited by superoxide dismutase. The horseradish peroxidase-catalyzed oxidation of isoniazid produced two different carbon-centered radicals. The identity of these radicals is not clear; however, they may arise from an acyl (RCO) radical and an alkyl (R) radical. PMID- 6296082 TI - Expression and partial DNA sequence of the chicken beta H-globin gene. AB - We have determined the DNA sequence of the 3' end of the putative beta H-globin gene of chickens and used an S1 nuclease protection assay to analyze transcription of this gene. Appearance of mRNA corresponding to the putative beta H-globin gene in the erythroid cells of developing chicken embryos coincides with the reported expression of the beta H-globin protein, a constitutent of hemoglobin H which is a minor component in the definitive cells of late embryos and newly hatched chickens, thereby confirming the tentative identification of this gene as beta H. Analysis of the DNA sequence of the 3'-flanking region of the beta H gene revealed the presence of a complex array of direct repeats. A part of this "repeat cluster" is homologous to an inserted element in the large intron of the goat beta A- and beta C-globin genes. PMID- 6296083 TI - Transformation by Rous sarcoma virus induces similar patterns of glycosaminoglycan synthesis in chick embryo skin fibroblasts and vertebral chondroblasts. AB - Chick embryo skin fibroblasts and vertebral chondroblasts were infected with a temperature-sensitive mutant of Rous sarcoma virus, LA24A, and were grown at permissive (36 degrees C) and nonpermissive (41 degrees C) temperatures. During exponential growth, infected and parallel uninfected cultures were labeled with D [3H]glucosamine, and newly synthesized glycosaminoglycans were identified by anion exchange chromatography and by selective enzymatic and chemical degradations. Control fibroblasts synthesized low levels of hyaluronic acid (HA), and dermatan sulfate (DS), moderate levels of heparan sulfate (HS), and high levels of chondroitin sulfate (CS). In contrast, control chondroblasts synthesized very low levels of HA and DS, no HS, and very high levels of CS. Following transformation and growth at 36 degrees C, both cell types showed a dramatic increase in HA synthesis and a significant decrease in CS synthesis. In addition, transformed chondroblasts initiated the synthesis of HS and increased their synthesis of DS to levels that matched those of transformed fibroblasts. The CS chains synthesized by control chondroblasts were partially undersulfated, while those synthesized by both normal and transformed fibroblasts were fully sulfated. Upon transformation, chondroblasts grown at 36 degrees C initiated the synthesis of fully sulfated CS chains. Most of the above biosynthetic alterations were completely reversed when infected cells were grown at 41 degrees C, indicating that they were dependent on the transforming gene product of LA24A. Clearly, the profound differences that distinguish normal fibroblasts from normal chondroblasts are lost upon transformation, and these two types of terminally differentiated cells converge toward a common, though not identical, biosynthetic program for glycosaminoglycans. PMID- 6296084 TI - Regulation of phosphatidylcholine biosynthesis in mammalian cells. I. Effects of phospholipase C treatment on phosphatidylcholine metabolism in Chinese hamster ovary cells and LM mouse fibroblasts. AB - Addition of phospholipase C from Clostridium perfringens to cultures of Chinese hamster ovary (CHO) cells resulted in rapid degradation of cellular phosphatidylcholine with concomitant release of phosphocholine. The rate of incorporation of radiolabeled choline into lipids was increased 2-fold in phospholipase C-treated CHO cells as compared to untreated controls. The only enzyme in the pathway of phosphatidylcholine biosynthesis with increased activity in phospholipase C-treated cells was CTP:phosphocholine cytidylyltransferase, indicating that the cytidylyltransferase plays an important role in the stimulation of phosphatidylcholine biosynthesis. The phospholipase treatment was toxic to a CHO mutant cell line with abnormally low cytidylyltransferase activity. Mouse LM fibroblasts were resistant to enzymatic attack by phospholipase C, and cytidylyltransferase activity in LM cells did not change upon phospholipase C treatment. PMID- 6296085 TI - Regulation of phosphatidylcholine biosynthesis in mammalian cells. II. Effects of phospholipase C treatment on the activity and subcellular distribution of CTP:phosphocholine cytidylyltransferase in Chinese hamster ovary and LM cell lines. AB - CTP:phosphocholine cytidylyltransferase was located in both the cytosolic and particulate fractions from Chinese hamster ovary cells. The activity of the cytosolic form of the enzyme was greatly enhanced by incubation with sonicated preparations of several different lipids, although incubations with either phosphatidylcholine or 1,2-sn-diolein did not increase activity. The activation of the cytidylyltransferase in Chinese hamster ovary cells treated with phospholipase C from Clostridium perfringens occurred with a concomitant shift in the subcellular distribution of the enzyme from cytosolic to particulate fractions. This shift was rapid and did not require protein synthesis. Removal of phospholipase C from the cell cultures resulted in a return to basal levels of incorporation of [3H]choline into phosphatidylcholine, a decrease in the activity of cytidylyltransferase, and a loss of the membrane-bound form of the enzyme. Similar experiments with LM cells, which are resistant to exogenous phospholipase C, showed no change in subcellular distribution of cytidylyltransferase, suggesting that the activation of CTP:phosphocholine cytidylyltransferase required a change in membrane phospholipid composition. The results presented are discussed in terms of a mechanism of regulation of phosphatidylcholine production involving monitoring of membrane phospholipid composition. PMID- 6296086 TI - Regulation of phosphatidylcholine biosynthesis in mammalian cells. III. Effects of alterations in the phospholipid compositions of Chinese hamster ovary and LM cells on the activity and distribution of CTP:phosphocholine cytidylyltransferase. AB - The activity and subcellular distribution of CTP:phosphocholine cytidylyltransferase in LM and Chinese hamster ovary cells in which the phospholipid composition had been altered by supplementary feeding with choline analogues were examined. Decreased levels of cellular phosphatidylcholine with corresponding increased levels of either phosphatidylethanolamine, phosphatidylmonomethylethanolamine, or phosphatidyldimethylethanolamine resulted in increased CTP:phosphocholine cytidylyltransferase activity in cell homogenates. In addition, a significantly larger fraction of the total cytidylyltransferase activity was membrane-bound in these cells. The activity of the cytidylyltransferase from cytosolic extracts of both cell types was found to be greatly increased when assayed in the presence of either phosphatidylmonomethyl ethanolamine or phosphatidyldimethylethanolamine. The lysophosphatide forms of these lipids were found to be poor activators of the cytidylyltransferase. These findings suggest that the regulation of phosphatidylcholine biosynthesis is at least partially dependent on information transfer from membranes to CTP: phosphocholine cytidylyltransferase. That is, in the presence of phosphatidylcholine-deficient membranes of cytidylyltransferase becomes activated and associated with the membranes. PMID- 6296087 TI - Biological role of epidermal growth factor-receptor clustering. Investigation with monoclonal anti-receptor antibodies. AB - Monoclonal anti-epidermal growth factor (EGF) receptor antibodies were used as a diagnostic tool for the investigation of the role of the receptor molecule in the transduction of the biological effects mediated by EGF. The specificity of the antibodies was established by immunoprecipitation of the receptor from biosynthetically labeled cells. The previously described (Schreiber, A. B., Lax, I., Yarden, Y., Eshhar, Z., and Schlessinger, J. (1981) Proc. Natl. Acad. Sci U. S. A. 78, 7535-7539) 2F2-IgM antibody binds to or close to the binding site of the growth factor on the receptor molecule; it induces receptor clustering and internalization and triggers early and delayed effects of EGF. A monovalent Fab fragment of the 2G2-IgM antibody stimulates the EGF-receptor sensitive protein kinase, but does not induce receptor clustering and DNA synthesis. When cell bound 2G2-Fab fragments are cross-linked with anti-mouse Ig antibodies, both receptor clustering and mitogenic activity are restored. A second monoclonal antibody against EGF-receptor denoted TL5-IgG does not interfere with the binding of EGF to the receptor, fails to induce receptor clustering, and does not possess any intrinsic bioactivity. The cross-linking of cell-bound TL5-IgG with anti mouse Ig antibodies leads to the stimulation of DNA synthesis. It is concluded that the EGF-receptor, when properly triggered, contains all the biochemical attributes necessary for the initiation of biological effects. Receptor clustering, even when mediated via domains distinct from the hormone binding site on the receptor molecule, appears as a necessary and sufficient signal for the induction of DNA synthesis. PMID- 6296088 TI - Specific receptor sites for pyridoxal 5'-phosphate and pyridoxal 5' deoxymethylenephosphonate at the alpha and beta NH2-terminal regions of hemoglobin. AB - hemoglobins have been prepared which are substituted with pyridoxal 5'-phosphate or pyridoxal 5'-deoxymethylenephosphonate at both NH2-terminal amino groups of either the alpha or the beta chains. 31P NMR titration curves of the 5' side chain show a substantial decrease in acid strength in hemoglobins pyridoxylated at the alpha chains and the reverse effect in the hemoglobins labeled in the beta chains. These changes can readily be explained by the interaction of the 5' side chains with surrounding residues which provide specific binding sites for the monoanion at the alpha chain termini and for the dianion in the case of the beta chains. Since the proportion of monoanion is at lest ten times greater for pyridoxal 5'-deoxymethylenephosphonate than for pyridoxal 5'-phosphate at neutral pH, the phosphonate reacts preferentially with the alpha chain NH2-terminal amino groups and the phosphate with the beta chain ones. PMID- 6296089 TI - Characterization of a glycoprotein alpha-galactosidase from lentil seeds (Lens culinaris). AB - alpha-Galactosidase (alpha-D-galactoside galactohydrolase, EC 3.2.1.22), an enzyme responsible for mobilizing the raffinose family of oligosaccharides in legume seeds, has been isolated from lentils (Lens culinaris) and purified about 4,000-fold. The Sephadex gel filtration profile showed the presence of two forms of the enzyme, alpha-galactosidase I with an apparent Mr = 160,000 and alpha galactosidase II of Mr = 40,000. Enzyme II readily aggregates to form I when any attempt is made to concentrate the solution. Thus, only enzyme I was purified and its properties studied. The multistep purification procedure included affinity binding of the enzyme to concanavalin A-Sepharose, indicating its glycoprotein nature with glucose/mannose termini of the carbohydrate moieties. The amino acid and carbohydrate compositions of the native enzyme and that of the glycopeptide obtained from pronase-digested, denatured enzyme have been determined. Asparagine seems to be involved in forming the linkage with the carbohydrate moiety. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of enzyme I shows a single protein band with Mr = 40,000 which also stains with periodic acid-Schiff reagent. Thus, the enzyme consists of four identical glycoprotein subunits. The isoelectric point of the enzyme is 8.0. The pH optima of enzyme I and II are 6.1 and 4.7, respectively. The substrate specificity and the mode of substrate inhibition of enzyme I is discussed. The effect of temperature on Vmax and Km of the enzyme is presented; at pH 6.1 the energy of activation is 62.1 kJ/mol and the delta H value is -34.3 kJ/mol in the temperature range 20-50 degrees C. Preliminary studies show that enzyme I possesses hemagglutinating properties with glucose/mannose specificity. PMID- 6296090 TI - Pure yeast RNA polymerase B (II) initiates transcription at specific points on supercoiled yeast DNA. AB - Pure yeast RNA polmymerase B (II) can selectively initiate abortive transcription on a supercoiled recombinant plasmid DNA carrying yeast DNA in the presence of low concentrations of ribonucleoside triphosphates and Mn2+. Five major products ranging between 60 and 150 nucleotides were characterized by hybridization. Three of them originate from the vector pBR322 and two from the yeast DNA insert. Based on a RNA primer extension reaction with recombinant M13 DNAs as template, a method allowing the mapping of the short abortive RNA products has been developed. An initiation site within the yeast DNA insert has thus precisely been mapped. The DNA sequence in this region was determined and showed several relevant features. The in vitro initiation site is preceded by a potential TATATATA box at -40 base pairs and at -105 by the sequence GTTAATCT similar to the consensus sequence GCTCAATCT usually found around 80 base pairs upstream from the cap site. Large blocks of alternated purine pyrimidine residues are found in this region as for several known yeast promotors. The 5' end of the RNA initiated from this site contains several potential signals for the initiation of translation. The possibility that a B to Z transition of DNA could be important for the interaction of the RNA polymerase with its template is discussed. PMID- 6296091 TI - Studies on the hysteretic properties of chloroplast fructose-1,6-bisphosphatase. AB - Chloroplast fructose-1,6-bisphosphatase hysteresis in response to modifiers was uncovered by carrying out the enzyme assays in two consecutive steps. The activity of chloroplast fructose-1,6-bisphosphatase, assayed at low concentrations of both fructose-1,6-bisphosphatase and Mg2+, was enhanced by preincubating the enzyme with dithiothreitol, thioredoxin f, fructose 1,6 bisphosphate, and Ca2+. In the time-dependent activation process, fructose 1,6 bisphosphate and Ca2+ could be replaced by other sugar biphosphates and Mn2+, respectively. Once activated, chloroplast fructose-1,6-bisphosphatase hydrolyzed fructose 1,6-bisphosphate and sedoheptulose 1,7-bisphosphate in the presence of Mg2+, Mn2+, or Fe2+. The A0.5 for fructose 1,6-bisphosphate (activator) was lowered by reduced thioredoxin f and remained unchanged when Mg2+ was varied during the assay of activity. On the contrary, the S0.5 for fructose 1,6 bisphosphate (substrate) was unaffected by reduced thioredoxin f and depended on the concentration of Mg2+. Ca2+ played a dual role on the activity of chloroplast fructose-1,6-bisphosphatase; it was a component of the concerted activation and an inhibitor in the catalytic step. Provided dithiothreitol was present, the activating effectors were not required to maintain the enzyme in the active form. Considered together these results strongly suggest that the regulation of fructose-1,6-bisphosphatase in chloroplast occurs at two different levels, the activation of the enzyme and the catalysis. PMID- 6296092 TI - Localization of membrane proteins by the use of a photoreactive fatty acid incorporated in vivo into vesicular stomatitis virus. AB - Vesicular stomatitis virus grown in the presence of omega-[9-3H]diazirinophenoxy nonanoate resulted in the biosynthetic incorporation of this photoreactive fatty acid into viral phospholipids as well as into the membrane anchoring domain of the viral G glycoprotein. Photolysis of the isolated virus at 360 nm resulted in extensive labeling of the G protein but none of the other proteins by the viral phospholipids. In addition, a new product was obtained and was identified as a G G dimer by its molecular weight and its reaction with anti-G antibody. These results demonstrate the use of photoreactive fatty acid to identify integral membrane proteins and to make photoaffinity probes of fatty acylated membrane proteins. PMID- 6296093 TI - Nucleotide sequences of two pea cDNA clones encoding the small subunit of ribulose 1,5-bisphosphate carboxylase and the major chlorophyll a/b-binding thylakoid polypeptide. AB - Two major chloroplast proteins are encoded by nuclear genes and synthesized on free cytoplasmic ribosomes: the small subunit of ribulose 1,5-bisphosphate carboxylase and the apoprotein components of the chlorophyll a/b light harvesting complex. We have recently reported the isolation of two cDNA clones from pea which encode both the small subunit of ribulose 1,5-bisphosphate carboxylase (pSS15) and the polypeptide 15 (pAB96), the major chlorophyll a/b binding protein (Broglie, R., Bellemare, G., Bartlett, S., Chua, N.-H., and Cashmore, A. R. (1981) Proc. Natl. Acad. Sci. U.S.A. 78, 7304-7308). To further characterize these clones, we determined their nucleotide sequence. Clone pSS15 contains a 691 base pair cDNA insert which encodes the entire 123 amino acids of the mature small subunit protein. In addition, this clone also encodes 33 amino acids of the NH2-terminal transit peptide extension and 148 nucleotides of the 3' noncoding region preceding the poly(A)tail. A second cDNA clone (pAB96) contains an 833 nucleotide insert which encodes most of polypeptide 15. The DNA sequence of this cloned cDNA was used to deduce the previously undetermined amino acid sequence of this integral thylakoid membrane protein. The nucleotide sequence of the cDNA clone, pSS15, should provide information concerning the role of the transit sequence in the transport of cytoplasmically synthesized chloroplast proteins. Similarly, the deduced amino acid sequence of polypeptide 15 will provide information for predicting its orientation in thylakoid membranes as well as its role in binding chlorophyll. PMID- 6296094 TI - Opiate receptor binding affected by guanine nucleotide. Partial loss of sensitivity to guanosine-5'-(beta,gamma-imido)triphosphate evident in vitro. AB - The sensitivity of the receptor dissociation rate of the opiate agonist [3H]etorphine to the effects of guanosine-5'-(beta, gamma-imido)triphosphate and Na+ was measured in washed rat brain membrane homogenates after in vivo labeling. Comparison to the previously measured rapid in vivo dissociation curve (t1/2 approximately 50 s) (Perry, D. C., Rosenbaum, J. S., Kurowski, M., and Sadee, W. (1982) Mol. Pharmacol. 21, 272-279) revealed that brain homogenization and membrane washing procedures significantly prolonged the dissociation rate, even when measured in the presence of Na+ and guanyl nucleotide. The in vivo dissociation rate could only be reproduced in vitro when labeling occurred in vivo and brain homogenization occurred in the presence of these regulatory factors. The prolonged in vitro [3H]etorphine dissociation curve was predominately a result of a decreased sensitivity to guanine nucleotide rather than to Na+. These data suggest that partial functional uncoupling of the opiate receptor-effector system may occur in vitro. PMID- 6296095 TI - Stereospecific elimination of hydrogen at C-10 in eicosapentaenoic acid during the conversion to leukotriene C5. AB - (10L)- and (10D)-[1-14C, 10-3H]5,8,11,14,17-eicosapentaenoic acids were synthesized to investigate mechanistic and stereochemical aspects of leukotriene biosynthesis. Experiments with mastocytoma cells showed that a hydrogen is stereospecifically eliminated from C-10 during the conversion of eicosapentaenoic acid to leukotriene C5. The hydrogen lost has the pro-S (D) configuration. 5 Hydroxy-6,8,11,14,17-eicosapentaenoic acid, formed in the same experiments, was enriched in tritium when the (10D), but not when the (10L), isomer of labeled eicosapentaenoic acid was used. This indicates that oxygenation of the acid at C 5 occurred before the elimination of hydrogen and suggests that removal of the pro-S hydrogen at C-10 in 5-hydroperoxy-6,8,11,14,17-eicosapentaenoic acid initiates its transformation to trans-5(S),6(S)-oxido-7,9-trans-11,14,17-cis eicosapentaenoic acid (leukotriene A5). PMID- 6296096 TI - Separation and identification of type 1 and type 2 protein phosphatases from rabbit reticulocyte lysates. AB - Protein phosphatase type 1 and type 2 activities (designated PP-1 and PP-2, respectively) from rabbit reticulocyte lysates have been identified and characterized based on criteria previously established for similar activities in rabbit skeletal muscle and rabbit liver. These include (a) chromatographic separation on DEAE-cellulose, (b) substrate specificity toward glycogen phosphorylase a and the alpha- and beta-subunits of phosphorylase kinase, (c) differential sensitivity to the heat-stable protein phosphatase inhibitors-1 and 2, and (d) sensitivity to MgATP. When total lysate phosphatases are assayed in the presence of 1 mM MnCl2, protein phosphatase type 2 represents 84% of lysate phosphorylase phosphatase activity. However, when phosphatase assays are carried out with MgATP concentrations similar to those in the lysate, type 2 activity is diminished, and the levels of type 1 (41%) and type 2 (59%) phosphatase activities are comparable. A small proportion (6%) of total lysate phosphatase is tightly bound to the ribosomes, where type 1 phosphatase predominates. At least five species of protein phosphatases can be identified in lysates. These constitute two forms of protein phosphatase type 1, one of which (designated FC) is dependent on MgATP and a lysate activator protein FA; both FC and FA have been identified previously in skeletal muscle. Three species of protein phosphatase type 2 have been identified and designated PP-2B, PP-2A1, and PP-2A2 based on criteria recently established for rabbit skeletal muscle and rabbit liver phosphatases, which display similar phosphatase profiles. Lysate protein phosphatases types 1, FC, 2A1, and 2A2 can all act on phosphorylase a and the alpha- (type 2) or beta-(type 1) subunit of phosphorylase kinase. PP-2B, a Ca2+/calmodulin-dependent phosphatase, specifically dephosphorylates the alpha subunit of phosphorylase kinase, but does not act on phosphorylase alpha. The heat-stable protein phosphatase inhibitor-2 from skeletal muscle completely blocks the activity of the two type 1 phosphatases (PP-1, FC), but has no effect on the three species of type 2 protein phosphatase. A preliminary assay of the two heat-stable phosphatase inhibitors in lysates indicates significant levels of inhibitor-2, but little or no detectable inhibitor-1. PMID- 6296097 TI - GTP-dependent anion-sensitive adenylate cyclase in snail ganglia potentiation of neurotransmitter effects. AB - Snail ganglia possess an anion-sensitive adenylate cyclase. This enzyme was stimulated 100% by chloride in a strictly GTP-dependent manner. The apparent affinity of chloride for adenylate cyclase was 2 X 10(-4) M. Halogens were found to be the most active anions. Some inorganic anions such as SO4(2-) and H2PO4- were inactive, as were all the organic anions tested. Stimulation was not cumulative for any maximal concentration of the active anions except fluoride. Chloride potentiated the effect of fluoride, indicating that the anion effect is not fluoride-like. Another striking result is that chloride enhanced adenylate cyclase sensitivity to the neurotransmitters serotonin and dopamine. The absence of chloride stimulation when Mg2+ was replaced by Mn2+ further indicates a role of the GTP-binding protein (the G/F unit). Chloride could reversibly stimulate the adenylate cyclase activity already maximally stimulated by guanyl 5' imidodiphosphate. We therefore suggest that, in snail ganglia, chloride raises the activity of the G/F unit-catalytic unit complex at some stage after its formation. The same specific anion-sensitive adenylate cyclase was also found in some of the rat tissues tested. PMID- 6296098 TI - Measurement of the electrochemical proton gradient in submitochondrial particles. AB - The pH gradient and membrane potential of submitochondrial particles from bovine heart were estimated by the uptake of [14C]ethylamine and [36Cl]perchlorate, using filtration through a glass fiber prefilter and Millipore filter without washing to separate the vesicles from the medium. An external volume probe of [3H] sucrose was also used. Internal volume of the vesicles was measured by the extent of uptake of glucose, which equilibrates slowly across the membrane. The electrochemical potential gradient of H+ (delta micro H+) calculated from uptake of ethylamine and perchlorate, assuming the ions taken up were free in solution inside the vesicles, was 23 to 24 kJ/mol of H+ (240-250 mV) during respiration in the absence of ATP. The ratio of the free energy of ATP synthesis (delta GATP) to delta micro H+ was 2.2 to 2.3 during oxidative phosphorylation and only slightly higher during ATP hydrolysis indicating that the H+-translocating ATPase is close to equilibrium under both conditions. The nonintegral ratio suggests there is a systematic error in the measurement of delta micro H+. The value of delta micro H+ calculated from ion uptake could be too high if some of the ions taken up are bound to the membrane or concentrated into the electric double layer at the inner membrane-water interface. The effects of vesicle volume (varied osmotically) and permeant ions (which affect internal ionic strength and pH) on the ratio of delta GATP to delta micro H+ suggested that ion association with the membrane in fact caused significant overestimation of delta micro H+. Association of ethylammonium and perchlorate ions with unenergized submitochondrial particles was measured by centrifugation, in the presence of a high concentration of impermeant salt to minimize association with the external surface. The results were used to estimate the extent of binding during the ion uptake assays, and delta micro H+ was recalculated taking this binding into account. The resulting values were between 19 and 20 kJ/mol of H+ (197-207 mV) during respiration in the absence of ADP, and the ratio of delta GATP to delta micro H+ was about 3 during oxidative phosphorylation. PMID- 6296099 TI - The action of insulin on hepatic fructose 2,6-bisphosphate metabolism. AB - The inhibition of hepatocyte 6-phosphofructo-1-kinase by glucagon was suppressed by insulin when the enzyme was measured in crude extracts. However, no effect of either hormone was observed after the removal of allosteric effectors from the enzyme, suggesting that the alterations in activity may be due to changes in the level of fructose 2,6-bisphosphate, a potent allosteric activator of the enzyme. Insulin opposed the action of both glucagon and exogenous cyclic AMP to lower fructose 2,6-bisphosphate levels. The concentration of glucagon and of cyclic AMP that gave a half-maximal decrease in fructose 2,6-bisphosphate levels was increased in the presence of 10 nM insulin from 0.03 to 0.09 nM and from 12 to 36 microM, respectively. Insulin also counteracted the effect of maximal concentrations of epinephrine on fructose 2,6-bisphosphate levels. In the presence of 0.02 nM glucagon or 10 microM epinephrine, 10 nM insulin enhanced 6 phosphofructo-2-kinase and decreased fructose 2,6-bisphosphatase activity in (NH4)2SO4-treated hepatocyte extracts. The bifunctional enzyme 6-phosphofructo-2 kinase/fructose 2,6-bisphosphatase was shown to be a substrate for the cAMP dependent protein kinase but not for phosphorylase kinase. It was concluded that insulin opposed the action of glucagon and epinephrine by affecting the phosphorylation state of 6-phosphofructo-2-kinase/fructose 2,6-bisphosphatase. Fructose 2,6-bisphosphate levels were decreased in liver cells from diabetic rats. Addition of 30 mM glucose elevated fructose 2,6-bisphosphate levels in cells from fed and 24-h-starved rats but not in cells from diabetic rats. This was probably due to decreases in both 6-phosphofructo-2-kinase and glucokinase activity in the diabetic state. These results show that insulin has both short and long term effects on fructose 2,6-bisphosphate metabolism in liver. PMID- 6296100 TI - Turnover of cytochrome c oxidase from Paracoccus denitrificans. AB - The heme aa3 type cytochrome oxidase from Paracoccus denitrificans incorporated into vesicles with phospholipid reacts during turnover much as the oxidase from mitochondria does. The spectrophotometric changes observed at various wavelengths are closely similar, and the rate is about one-half of that for beef heart oxidase under the same conditions. The rate of appearance of oxidized cytochrome c on initiation of the reaction is also similar and depends on the previous treatment of the oxidase as described by Antonini, E., Brunori, M., Colosimo, A., Greenwood, C. and Wilson, M. T. (1977) Proc. Natl. Acad. Sci. U.S.A. 74, 3128 3132. In terms of their model the resting Paracoccus enzyme is converted to the pulsed form during turnover. The effect is observed with both cytochrome c and hexamine ruthenium as reductants. With the latter a 60-fold increase in rate is observed. PMID- 6296101 TI - NADPH-dependent production of oxy radicals by purified components of the rat liver mixed function oxidase system. I. Oxidation of hydroxyl radical scavenging agents. AB - Isolated microsomes catalyze an NADPH-dependent oxidation of typical hydroxyl radical scavenging agents. To determine which microsomal components participate in the oxidation of the scavengers, experiments were carried out with purified NADPH-cytochrome P-450 reductase and cytochrome P-450 isolated from phenobarbital treated rats. The production of ethylene from 4-ketothiomethylbutyrate or of formaldehyde from either dimethyl sulfoxide or tertiary butyl alcohol was measured in the presence of NADPH plus reductase or NADPH plus reductase plus cytochrome P-450. The reductase-dependent system itself catalyzed the oxidation of the three scavengers. Addition of cytochrome P-450 had no effect on the rates of oxidation of the scavengers. Relative rates of oxidation of the scavengers reflected the amount of the reductase in the assay system. Varying the amount of cytochrome P-450 at a fixed concentration of the reductase did not result in rates different from that observed in the absence of cytochrome P-450. Dilauroyl phosphatidylcholine appeared to have a stimulatory role in these oxidations, whereas cytochrome P-450 reductase and NADPH were obligatory components. The reductase-dependent oxidation of the scavengers was inhibited by superoxide dismutase, catalase, and competing hydroxyl radical scavenging agents suggesting that superoxide, hydrogen peroxide, and an oxygen radical with the oxidizing power of the hydroxyl radical played a role in these oxidations. Further, these oxidations were inhibited by the iron-chelator desferrioxamine but stimulated by either EDTA or by iron. These results suggest that an iron-catalyzed Haber-Weiss reaction might be involved in the mechanism by which purified cytochrome P-450 reductase mediates the oxidation of typical hydroxyl radical scavengers. The results demonstrated that NADPH-cytochrome P-450 reductase may represent an important locus of oxygen activation leading to the production of a highly oxidizing species characteristic of the hydroxyl radical. PMID- 6296102 TI - NADPH-dependent production of oxy radicals by purified components of the rat liver mixed function oxidase system. II. Role in microsomal oxidation of ethanol. PMID- 6296103 TI - Resolution and reassociation of three distinct components from pig heart phosphoprotein phosphatase. AB - A partially purified pig heart phosphoprotein phosphatase was dissociated into three distinct components, namely alpha, beta, and gamma, by gel filtration on Sephacryl S-200 followed by chromatography on DEAE-Sephadex in the presence of 6 M urea. Although alpha itself had phosphatase activities toward P-H2B histone, P H1 histone, phosphorylase a, and glycogen synthase b, beta and gamma had no activity toward these substrates even in the presence of 1 mM Mn2+. The beta component (Mr = 80,000) combined with alpha (Mr = 31,000) in the absence of urea to produce Form 2 (Mr = 123,000) with concomitant increase in P-H1 histone phosphatase activity and Mg2+ requirement for P-H2B histone phosphatase activity (Imazu, M., Imaoka, T., Usui, H., Kinohara, N., and Takeda, M. (1981) J. Biochem. 90, 851-862). The gamma component (Mr = 62,000) reassociated with Form 2 to produce Form 1 (Mr = 199,000) which was similar to the original phosphoprotein phosphatase in substrate specificity and Mg2+ requirement. Binding of gamma to Form 2 strongly suppressed the phosphatase activities toward phosphorylase a and glycogen synthase b with marginal effects on the other phosphatase activities and Mg2+ requirement. However, gamma alone could not associate with alpha. The gamma component was sensitive to treatment with heat (60 degrees C for 2 min) or trypsin and was resistant to treatment with DNase or RNase. The pig heart phosphoprotein phosphatase was further purified to near homogeneity, as judged by polyacrylamide gel electrophoresis. Sodium dodecyl sulfate gel electrophoresis revealed that the purified enzyme (Mr = 171,000) was composed of three polypeptide components, namely alpha', beta', and gamma' with molecular weights of 34,000, 69,000, and 56,000, respectively. The component stoichiometry was determined to be alpha' 1 beta' 1 gamma' 1 by densitometric tracing of the Coomassie blue-stained bands on the acrylamide gel. After dissociation of alpha ' and other components by gel filtration of the purified enzyme on Sephacryl S-200 in the presence of 6 M urea, one alpha ' combined with one beta' to produce Form 2' of Mr = 106,000. Since Form 1 and the purified enzyme as well as Form 2 and Form 2' had similar catalytic properties and s20,w values, respectively, component compositions are suggested to be alpha 1 beta 1 gamma 1 for Form 1 and alpha 1 beta 1 for Form Form 2. PMID- 6296104 TI - 18O labeling of adenine nucleotide alpha-phosphoryls in platelets. Contribution by phosphodiesterase-catalyzed hydrolysis of cAMP. PMID- 6296105 TI - Thyrotropin-releasing hormone-diphtheria toxin-related polypeptide conjugates. Potential role of the hydrophobic domain in toxin entry. AB - We have separately coupled a modified thyrotropin-releasing hormone (TRH) molecule to two diphtheria toxin-related polypeptides, CRM26 and CRM45. Both polypeptides are enzymatically active but only CRM45 contains the hydrophobic domain of the toxin molecule. The TRH-CRM45 conjugate caused a 50% inhibition of protein synthesis in GH3 rat pituitary cells at 3 X 10(-9) M. CRM45 alone was 200 to 500 times less toxic than the conjugate. Intoxication by TRH-CRM45 was prevented by excess TRH, preincubation with diphtheria antitoxin, or reduction of the disulfide cross-link. In addition, TRH-CRM45 was no more toxic than CRM45 itself to 3T3 cells which lack TRH receptors. In contrast, TRH-CRM26, although it retained enzymatic activity, was nontoxic for GH3 cells even at 10(-7) M. Binding experiments showed that both conjugates compete for TRH receptors with comparable affinities. The potential role of the hydrophobic domain in toxin entry is discussed. PMID- 6296106 TI - Inhibition of electron transfer by 3-alkyl-2-hydroxy-1,4-naphthoquinones in the ubiquinol-cytochrome c oxidoreductases of Rhodopseudomonas sphaeroides and mammalian mitochondria. Interaction with a ubiquinone-binding site and the Rieske iron-sulfur cluster. AB - 3-Alkyl-2-hydroxy-1,4-naphthoquinones (alkyl-HNQ) inhibit Rieske iron-sulfur cluster (Rieske FeS) oxidation and cytochrome b reduction in ubiquinol-cytochrome c oxidoreductase. The effects are the same as those of 5-undecyl-6-hydroxy-4,7 dioxobenzothiazole. Concentrations for 50% inhibition in chromatophores of Rhodopseudomonas sphaeroides (at 0.4 microM reaction center) are 2 microM for undecyl-, 3 microM for octyl-, and 40 microM for pentyl-substituted hydroxynaphthoquinones. The ethyl-substituted and unsubstituted derivatives do not inhibit electron transfer below 2 mM. In chromatophores in which the ubiquinone is partially extracted by isooctane (leaving 4 ubiquinones/reaction center), undecyl-HNQ is effective at 2.5 times lower concentration than in normal chromatophores (30 ubiquinones/reaction center). This observation suggests that the binding of the inhibitor is competitive with ubiquinone. Undecyl-HNQ eliminates the effect that the ubiquinone redox state has on the line shape of the EPR signal of Rieske FeS. This supports the idea that alkyl-HNQ shares a common binding site with ubiquinone which is closely associated with Rieske FeS. The ubiquinone in question has a midpoint oxidation-reduction potential at pH 7 of 90 mV with a -60 mV/pH unit dependency. This value matches that of the ubiquinone pool rather than that of ubiquinone Z, which is functionally recognized as a component "between" cytochrome b and Rieske FeS. When Rieske FeS is oxidized, a 20 times higher concentration of undecyl-HNQ is required for the electron transfer inhibition. This is consistent with the observation that the binding of the inhibitor shifts the midpoint oxidation-reduction potential of Rieske FeS about 60 mV higher, which in turn means that the inhibitor binds about 10 times stronger to the site when Rieske FeS is reduced than when it is oxidized. The observations suggest that 3-alkyl-2-hydroxy-1,4-naphthoquinones inhibit electron transfer by acting as ubiquinone antagonists at a site closely associated with Rieske FeS. PMID- 6296107 TI - Interaction of ligands with the distal glutamine in elephant myoglobin. AB - The effects of distal glutamine (E7) replacement in elephant myoglobin were studied by comparing the temperature-dependent nitrosyl electron spin resonance spectra, redox potentials, and the acid-alkaline equilibria of elephant and human myoglobins. For myoglobins containing a distal histidine, the nitrosyl ESR spectra do not exhibit superhyperfine splitting until near liquid helium temperatures (Yoshimura, T., Ozaki, T., Shintani, Y., and Watanabe, H. (1979) Arch. Biochem. Biophys. 193, 301-313). Studies presented here show that the ESR spectra of nitrosyl elephant myoglobin exhibit 9-line superhyperfine splitting well above liquid nitrogen temperatures, similar to the temperature profiles of isolated heme complexes (Morse, R.H. (1980) Fed. Proc. 39, 2006). It is concluded that the shift in the spectral equilibrium to higher temperature indicates a diminished interaction between NO and the distal position in elephant myoglobin. In addition, the redox potential of elephant myoglobin was found to be nearly 100 mV greater than that of human myoglobin, and the pKa of the acid-alkaline equilibrium (oxidized myoglobin) was 8.5, being 0.4 unit less than that of other vertebrate myoglobins. These different reactivities between elephant and human myoglobins are discussed based on the nature of charge interactions between polar ligands and distal glutamine and histidine. PMID- 6296108 TI - Charge determination of proteins with polyelectrolyte titration. AB - A recently developed photometric version of polyelectrolyte titration was applied for the determination of the number of charged residues on globular proteins. Based on the observation that oppositely charged polyelectrolytes form, in general, stoichiometric polyelectrolyte complexes, the protein solutions were incubated in excess with an oppositely charged polyelectrolyte, and the residual amount was back-titrated using o-toluidine blue for end point detection. It was found that within the range pH 2 to pH 9 the interaction of the polyelectrolytes, potassium polyvinylsulfate, polydiallylammonium chloride, and N methylglycolchitosan iodide, with various proteins of known amino acid composition (ribonuclease A, trypsin, chymotrypsin A, pepsin, cytochrome c) occurs stoichiometrically through 1:1 ion pair interaction, irrespective of the spatial distribution of the interacting ionic sites. The close correspondence between the experimental data for the net charge and the calculated balance of ionized residues for the proteins at a given pH indicates that in the native structure of these proteins oppositely charged ionic functions are largely neutralized by the formation of intramolecular salt linkages. It is concluded that polyelectrolyte titration offers an easy access to the determination of the surface charge of proteins and other biopolymers. The data further support the notion of the importance of electrostatic cooperative interactions in biological systems. PMID- 6296109 TI - Cordycepin analogues of 2-5A and its derivatives. Chemical synthesis and biological activity. AB - Cordycepin (3'-deoxyadenosine) analogues of 2-5A were prepared by dicyclohexylcarbodiimide-induced polymerization of cordycepin 5'-monophosphate. A series of oligomers of the general formula p5'(3'dA)-2'[p5'(3'dA)]n (n = 1-5) was obtained. Cordycepin trimer 5'-monophosphate (n = 2) and cordycepin tetramer 5' monophosphate (n = 3) were converted to the corresponding 5'-di- and triphosphates via the phosphorimidazolide method. Confirmation of assigned structures was provided through enzyme digestions, 1H and 31P NMR, and comparison with material which was synthesized by a completely independent route. Neither the cordycepin trimer triphosphate, ppp5'(3'dA)-2'p5'(3'dA)2'p5'(3'dA), nor the cordycepin tetramer triphosphate, ppp5'(3'dA)2'p5'(3'dA)2'p5'(3'dA)2'p5'-(3'dA), were inhibitors of translation in cell-free extracts of mouse L-cells programmed with encephalomyocarditis virus. In rabbit reticulocyte lysates, the cordycepin trimer triphosphate was devoid of activity, whereas the cordycepin tetramer triphosphate had approximately 1/100th the activity of 2-5A tetramer triphosphate, ppp5'A2'p5'A2'p5'A2'p5'A. Even though the cordycepin analogues were unable to activate the 2-5A-dependent endoribonuclease, they were able to bind to the endonuclease, thereby antagonizing the translational inhibitory effects of 2 5A. Thus, replacement of the 3'-hydroxyl groups of all the ribose moieties of 2 5A results in an analogue which can bind to the 2-5A-dependent endoribonuclease but is incapable of activating the enzyme. The 3'-hydroxyl groups of 2-5A, therefore, are critical for activation of the 2-5A-dependent endonuclease. PMID- 6296110 TI - Stereochemical course of DNA hydrolysis by nuclease S1. AB - Nuclease S1 hydrolyzes the Sp-diastereomer of 5'-O-(2'-deoxyadenosyl)-3'-O thymidyl phosphorothioate in H2(18)O to [18O]deoxyadenosine 5'-O-phosphorothioate which can be phosphorylated enzymatically to the Sp-diastereomer of [alpha 18O]deoxyadenosine 5'-O-(1-thiotriphosphate). 31P nmr spectroscopy shows the oxygen-18 in this compound to be in a nonbridging position at the alpha phosphorus, indicating that the hydrolysis reaction catalyzed by nuclease S1 proceeds with inversion of configuration at phosphorus. This result is compatible with a direct nucleophilic attack of H2O at phosphorus without the involvement of a covalent enzyme intermediate. PMID- 6296111 TI - Diacylglycerol kinase from pig brain. Purification and phospholipid dependencies. AB - Diacylglycerol kinase (EC 2.7.1.-) was purified 1,650-fold from pig brain cytosol. The purified enzyme showed a single protein band on polyacrylamide gel electrophoresis in the presence and absence of sodium dodecyl sulfate. The molecular weight of the kinase was estimated to be 78,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. A similar value (76,000) was obtained by Sephadex G-150 gel filtration. The activity of the purified enzyme was markedly enhanced by either deoxycholate or phospholipids. The extent of activation by phospholipids was in the order of phosphatidylcholine greater than lysophosphatidylcholine greater than phosphatidylethanolamine approximately equal to phosphatidylserine greater than sphingomyelin. Other phospholipids and unsaturated fatty acids were ineffective. Phosphatidylcholines from egg yolk and pig brain, and dioleoyl phosphatidylcholine were similarly effective. Saturated phosphatidylcholines with acyl chain lengths shorter than palmitate also gave a considerable activation. The activity with phosphatidylcholine was from 1.5- to 2.5-fold higher than that measured with deoxycholate. A very small amount of phosphatidylinositol or phosphatidylglycerol potently inhibited the phosphatidylcholine-dependent (but not deoxycholate-dependent) kinase activity. The inhibition by phosphatidylinositol was varied according to its molar ratio to phosphatidylcholine. As little as about 2.5 mol per cent of phosphatidylinositol resulted in 50% inhibition of the phosphatidylcholine-dependent kinase activity. The deoxycholate- and phosphatidylcholine-dependent kinase activities showed almost the same Km values for the substrates. In both cases, the apparent Km values for ATP and diacylglycerol were 300 microM and about 60 microM, respectively. The kinase required Mg2+ for its activity. When compared to deoxycholate, phosphatidylcholine was more effective at higher Mg2+ concentrations. The deoxycholate-dependent activity showed a broad pH optimum at around 8.0, whereas the phosphatidylcholine-dependent activity formed a clear peak at pH 7.4. PMID- 6296112 TI - Proton nuclear magnetic resonance studies of the ligation states of the monomeric ferricytochrome c' from Rhodopseudomonas palustris. Modulation of axial histidine bonding via variable proton donation. AB - The monomeric ferricytochrome c' from Rhodopseudomonas palustris strain 37 has been examined by 1H NMR spectroscopy at 45 degrees C and 360 MHz and at 55 degrees C and 200 MHz in 2H2O. The pH-dependent characteristics of the spectra have been analyzed in terms of two pK values at approximately 6 and approximately 8, the latter of which appears to correspond to the previously observed transition between an acidic species and a neutral species for this class of proteins. Previously determined rate constants (J. T. Jackson, G. N. La Mar, R. G. Bartsch, and M. A. Cusanovich, manuscript in preparation) permit, for the first time, connection between the downfield hyperfine shifted resonances of the two forms by spectral simulation. Contrary to previous suggestions (Emptage, M. H., Xavier, A. V., Wood, J.M., Alsaadi, B. M., Moore, G. R., Pitt, R. C., Williams, R. J. P., Ambler, R. P., and Bartsch, R. G. (1981) Biochemistry 20, 58 64; La Mar, G. N., Jackson, J. T., and Bartsch, R., (1981) J. Am. Chem. Soc. 103, 4405-4410), the data indicate that no additional ligand is involved in the transition but that the bonding between the iron and the axial histidyl ligand is severely altered. It is shown that this is consistent with extensive imidazolate character for the axial ligand in the neutral form. PMID- 6296113 TI - De novo purine synthesis in human lymphocytes. Partial co-purification of the enzymes and some properties of the pathway. AB - A partially purified enzyme extract from lectin-transformed human peripheral blood lymphocytes synthesized purine nucleotides de novo. Although the relatively lower specific activity of the pathway compared with that in the avian liver preparation previously described (Rowe, P. B., McCairns, E., Madsen, G., Sauer, D., and Elliott, H. (1978) J. Biol. Chem. 253, 7711-7721) limited the extent of purification, a number of properties were established: (i) Ammonia could be utilized as readily as glutamine for the synthesis of phosphoribosylamine but only glutamine provided N-3 of the purine ring; (ii) in the presence of either GTP or NAD, AMP or GMP were synthesized; (iii) purine synthesis was inhibited at the level of phosphoribosylamine synthesis by both AMP and GMP, irrespective of whether ammonia or glutamine was the N donor; (iv) while the synthesis of AMP and GMP from IMP was self-regulated, GTP also appeared to be an inhibitor of the synthesis of GMP from IMP; (v) amidophosphoribosyltransferase was isolated from both transformed and nontransformed cells in a low molecular weight form which was converted to a high molecular weight form in the presence of GMP; and (vi) no evidence was obtained for the existence of a classical multienzyme complex for purine synthesis. PMID- 6296114 TI - Studies on adenosine triphosphate transphosphorylases. Human isoenzymes of adenylate kinase: isolation and physicochemical comparison of the crystalline human ATP-AMP transphosphorylases from muscle and liver. AB - Procedures are described for the isolation, in crystalline form, of the adenylate kinases from autopsy samples of human muscle and from human liver. Weight average molecular weights were determined by sedimentation equilibrium to be 22,000 (+/- 700) and 25,450 (+/- 160) for the human muscle and liver isoenzymes, respectively. By sodium dodecyl sulfate-polyacrylamide gel electrophoresis, their molecular weights were estimated to be 21,700 and 26,500 for the muscle and liver enzymes, respectively. Both isoenzymes are accordingly monomeric proteins in their native state. Amino acid analyses are reported here for the normal human liver, calf liver, and rabbit liver adenylate kinases and compared with the normal human muscle, calf muscle, and rabbit muscle myokinases. The liver types as a group and the muscle types as a group show a great deal of homology, but some distinct differences are evident between the liver and muscle enzyme groups, especially in the number of residues of His, Pro, half-cystine, and the presence of tryptophan in the liver enzymes. The normal human liver adenylate kinase, as isolated in this report, has proved to be similar in its properties, if not identical, to the adenylate kinase isolated directly from human liver mitochondria (Hamada, M., Sumida, M., Okuda, H., Watanabe, T., Nojima, M., and Kuby, S. A. (1982) J. Biol. Chem. 257, 13120-13128). Therefore, the liver-type adenylate kinase may be considered a mitochondrial type. PMID- 6296115 TI - Identification and expression of a cloned yeast heat shock gene. AB - We have isolated the yeast HSP90 gene which encodes the Mr = 90,000 heat shock inducible protein of this organism. When this gene is introduced into yeast on a multicopy plasmid vector, a dramatic increase is observed in the level of synthesis of the Mr = 90,000 heat shock-inducible protein. This protein overproduction is due to expression of the plasmid-borne HSP90 gene, which is under the same heat shock regulation as its chromosomal counterpart. The presence of an increased dosage of the HSP90 gene has no effect on the synthesis of the other major heat shock-inducible proteins and does not alter the heat shock associated phenotype of thermal tolerance. PMID- 6296116 TI - Transcriptional and translational start sites for the Bacillus thuringiensis crystal protein gene. AB - The nucleotide sequence of the promoter region and part of the coding region of the crystal protein gene from Bacillus thuringiensis var. kurstaki HD-1-Dipel has been determined by analysis of a recombinant plasmid from Escherichia coli. The start points for transcription of the gene in B. thuringiensis and in the E. coli strain carrying the recombinant plasmid were located by S1 nuclease mapping. Two adjacent start sites were identified using RNAs synthesized during sporulation of B. thuringiensis: transcription was initiated from one site early in sporulation and from the other site in the middle of sporulation. A good correlation was found between the appearance of the crystal protein gene-specific RNA and the production of the protein, indicating that the gene is primarily under transcriptional control during sporulation. Parallel studies with the recombinant strain of E. coli revealed the presence of only a single species of gene-specific RNA, regardless of the growth phase of the cells; the crystal protein was produced at all stages of growth. The sequence for eight amino acids at the NH2 terminus of the crystal protein was determined and the corresponding coding sequence was located in the DNA sequence. A potential ribosome binding site of 11 nucleotides was found, located three nucleotides upstream from the initiator ATG codon. The deduced sequence for the first 333 amino acids of the crystal protein is presented. PMID- 6296118 TI - Purification and characterization of yeast topoisomerase I. AB - Yeast topoisomerase I (Mr = 76,000) has been purified to 80% homogeneity using a combination of ion exchange, gel filtration, and DNA-cellulose chromatography. The enzyme was characterized with respect to its ability to relax supercoiled DNA and to catenate nicked circular DNA. Yeast topoisomerase I will remove both positive and negative turns in DNA supercoils in the absence of ATP and magnesium ion. The products of the catenating activity of the enzyme were examined on agarose gels and in the electron microscope. These analyses indicate that yeast topoisomerase I will generate large catenated DNA networks which appear to rearrange to multimeric linear structures upon long incubation time. PMID- 6296117 TI - Functional domains in introns. RNA processing intermediates in cis- and trans acting mutants in the penultimate intron of the mitochondrial gene for cytochrome b. AB - The penultimate intron of the split mitochondrial gene (cob) for apocytochrome b of Saccharomyces cerevisiae is of particular interest; it contains a long unassigned reading frame, is present in both long form (six exons) and short form (three exons) of the gene, and a product expressed from it is required for the removal of its transcript and that of an intron in the transcript of the oxi3 gene. Complementation analysis shows mutants in this intron to be either cis dominant or transrecessive. Cis-dominant mutants are located in the first third (approximately 350 base pairs) of the open and near the 3'-end of the closed reading frame, while trans-recessive mutants are scattered throughout the remaining two-thirds (approximately 750 base pairs) of the open frame. Mutants in both classes exhibit the same pattern of splicing defects in their transcripts, but for different reasons. Those in the trans-recessive class lack a functional maturase (probably a protein of Mr = 27,000) encoded wholly within the 3' terminal segment of the intron, and for this reason also fail to express oxi3. In contrast, cis-dominant mutants are incapable of providing the splicing complex with a substrate of appropriate 2 degrees structure. They also accumulate a novel transcript, 1900 nucleotides long, which contains the intron fused to the downstream (3') exons. This may reflect an inability of the splicing complex to complete the normal sequence of cleavage of the intron at its downstream junction and the ligation of the two exonic moieties. PMID- 6296119 TI - Properties of purified enzymes induced by pathogenic drug-resistant mutants of herpes simplex virus. Evidence for virus variants expressing normal DNA polymerase and altered thymidine kinase. AB - The DNA polymerases and thymidine kinases induced by three drug-resistant mutants of herpes simplex virus type 1 (S1, Tr7, and B3) and their common parent strain, SC16, have been purified and their properties compared. No significant differences were seen in the affinities of the polymerases for TTP and dGTP, or for the triphosphates of 9-(2-hydroxyethyloxymethyl)guanine (acyclovir) or (E)-5 (2-bromovinyl)-2'-deoxyuridine (BVdU) (drugs used in their isolation). In contrast all three mutants induced abnormal thymidine kinases. Those induced by the acyclovir-resistant mutants, S1 and Tr7, showed reduced affinities for thymidine, acyclovir, and also BVdU. Thymidine kinase induced by the BVdU resistant mutant B3 showed reduced affinity for BVdU, but its affinities for thymidine and acyclovir were similar to those of the wild type enzyme. Thus, it appears that these variants of herpes simplex virus express altered thymidine kinases with impaired ability to phosphorylate particular nucleoside analogue drugs and these characteristics probably account for the drug resistance of the viruses. This strategy for resistance is important as it may result in variants with undiminished pathogenicity. PMID- 6296120 TI - Epidermal growth factor and epidermal growth factor receptor-dependent phosphorylation of a Mr = 34,000 protein substrate for pp60src. AB - A Mr = 34,000 protein present in the 100,000 X g supernatant fraction from A431 human epidermoid carcinoma cells is the major radiolabeled phosphate acceptor from [gamma-32P]ATP in a cell-free system requiring epidermal growth factor (EGF) and EGF receptor kinase. This protein is immunoprecipitated by IgG directed against avian Mr = 34,000 cellular substrate for pp60src. Phosphoamino acid analysis of the Mr = 34,000 protein labeled with 32Pi from [gamma-32P]ATP in a cell-free system requiring EGF and EGF receptor kinase yielded radiolabeled phosphotyrosine with no detectable radioactivity in phosphoserine or phosphothreonine. PMID- 6296121 TI - Purification and properties of T4 phage thymidylate synthetase produced by the cloned gene in an amplification vector. AB - We have introduced the T4 thymidylate synthetase gene, resident in a 2.7-kilobase EcoRI restriction fragment, into an amplification plasmid, pKC30. By regulating expression of this gene from the phage lambda pL promoter within pKC30 in a thyA host containing a temperature-sensitive lambda repressor, the T4 synthetase could be amplified about 200-fold over that after T4 infection. At this stage, a 20 fold purification was required to obtain homogeneous enzyme, mainly by an affinity column procedure. The purified plasmid-amplified T4 synthetase appeared to be identical with the T2 phage synthetase purified from phage-infected Escherichia coli in molecular weight, amino end group analysis, and immunochemical reactivity. The individual nature of the phage and host proteins was revealed by the fact that neither the T2 nor the T4 enzyme reacted with antibody to the E. coli synthetase, nor did antibody to the phage enzymes react with the E. coli synthetase. These differences were corroborated by DNA hybridization experiments, which revealed the absence of apparent homology between the T4 and E. coli synthetase genes. The techniques and genetic constructions described support the feasibility of employing similar amplification methods to prepare highly purified thymidylate synthetases from other sources. PMID- 6296122 TI - Identification of the predominant substrate for ADP-ribosylation by islet activating protein. AB - Islet activating protein (IAP), a toxin isolated from Bordetella pertussis, blocks the ability of inhibitory hormones to attenuate adenylate cyclase activity and enhances the ability of stimulatory hormones to activate the enzyme. The toxin appears to act by catalyzing the transfer of ADP ribose from NAD to a 41,000-dalton protein in target cell membranes. A protein purified from rabbit liver membranes, apparently composed of 41,000- and 35,000-dalton subunits, is shown to be a specific substrate for IAP. Cholera toxin does not ADP-ribosylate this protein. In contrast, the purified guanine nucleotide-binding regulatory component of adenylate cyclase (G/F), which is ADP-ribosylated by cholera toxin, is not covalently modified by IAP. Equilibrium binding studies and photoaffinity labeling experiments demonstrate that the 41,000-dalton subunit of the IAP substrate has a specific binding site for guanine nucleotides. PMID- 6296123 TI - Thrombin-induced phosphodiesteratic cleavage of phosphatidylinositol bisphosphate in human platelets. AB - The addition of thrombin to human platelets prelabeled with 32Pi led to significant loss of radioactivity in phosphatidylinositol 4,5-bisphosphate within 5 s, followed by recovery or even increase by 2 min. Loss of label from phosphatidylinositol phosphate was much less marked. Stimulated loss of label from phosphatidylinositol was not seen, while labeled phosphatidate increased severalfold. The principal labeled water-soluble phosphates observed, in addition to 32Pi and [32P] ATP, co-migrated with inositol diphosphate and inositol triphosphate. This suggests that a pool of polyphosphoinositides is constantly undergoing phosphodiesteratic cleavage and resynthesis. Thrombin addition led to rapid increase in radioactivity in inositol triphosphate, but not in inositol diphosphate. We conclude that this early consequence of the thrombin-platelet interaction is the result of an increase in the phosphodiesteratic cleavage of phosphatidylinositol bisphosphate. PMID- 6296124 TI - Blockade of heterologous desensitization of prostate adenylate cyclase without blockade of homologous down regulation of receptors or loss of GTP regulation of agonist binding. AB - Exposure of rat prostatic tissues to isoproterenol resulted in rapid desensitization of their catecholamine-sensitive adenylate cyclase which was associated with reduction of available beta-adrenoceptors and a loss of guanine nucleotide-mediated regulation of agonist binding to these receptors. The effect of isoproterenol treatment on responsiveness of the adenylate cyclase was prevented by acetylcholine, high potassium ion, or the calcium ionophore A23187. Preservation of responsiveness was not accompanied by maintenance of available beta-adrenoceptors or maintenance of guanine nucleotide regulation of agonist bindings. These results suggest that the lesion of the guanine nucleotide regulating components coupled to the catalytic moiety of the enzyme complex is a crucial factor in the desensitization of catecholamine-sensitive adenylate cyclase and the preservation of enzyme reaction could be accomplished by agents increasing intracellular calcium, which in turn, maintain the nucleotide regulatory components coupling to the cyclase in a protective environment from desensitization. PMID- 6296125 TI - Norepinephrine causes alpha 1-adrenergic receptor-mediated decrease of phosphatidylinositol in isolated rat liver plasma membranes supplemented with cytosol. AB - When purified rat liver plasma membranes were incubated with norepinephrine, rat liver cytosol, and Ca2+, the amount of membrane-bound phosphatidylinositol was reduced by up to 50%. The levels of other major membrane phospholipids underwent negligible change. The decrease in phosphatidylinositol levels was not observed if norepinephrine was omitted or cytosol was absent. The disappearance of phospholipid persisted when soluble Ca2+ was depleted by ethylene glycol bis(beta aminoethyl ether)-N,N,N',N'-tetraacetic acid. The decrease was prevented by phentolamine, benextramine, and prazosin, but not by sotalol. The results show that norepinephrine elicits a specific alpha 1-adrenergic receptor-mediated breakdown of phosphatidylinositol in isolated plasma membranes which is dependent on the presence of cytosol. PMID- 6296126 TI - Octahedral metal coordination in the active site of glyoxalase I as evidenced by the properties of Co(II)-glyoxalase I. AB - Co(II)-glyoxalase I has been prepared by reactivation of apoenzyme from human erythrocytes with Co2+. The visible absorption spectrum showed maxima at 493 and 515 nm and shoulders at 465 and 615 nm. The absorption coefficients at 493 and 515 nm were 35 and 33 M-1 cm-1/cobalt ion, respectively; i.e. 70 and 66 M-1 cm-1 for the dimeric metalloprotein. The product of the enzymatic reaction, S-D lactoylglutathione, although binding to Co(II)-glyoxalase I, had no demonstrable effect on the visible absorption spectrum, indicating binding outside the first coordination sphere of the metal. The EPR spectrum at 3.9 K was characterized by g1 approximately 6.6, g2 approximately 3.0, and g3 approximately 2.5, and eight hyperfine lines with A1 = 0.025 cm-1. Binding of the strong competitive inhibitor S-p-bromobenzylglutathione to Co(II)-glyoxalase I gave three g values: 6.3, 3.4, and 2.5, indicating a conformational change affecting the environment of the metal ion. Both optical and EPR spectra strongly suggest a high spin Co2+ with octahedral coordination in the active site of the enzyme. The similarities in kinetic properties between native Zn(II)-glyoxalase I and enzyme substituted with Mg2+, Mn2+, or Co2+ is consistent with the view that these enzyme forms have the same metal coordination in the protein. PMID- 6296127 TI - Identification and significance of glucokinase in transplantable insulinomas. AB - Glucose 6-phosphotransferases were investigated in two transplantable rat insulinoma tumor lines. Homogenates of tumors contained glucose phosphorylating activities of both high (e.g. supernatant Km = 0.060 mM and pellet Km = 0.077 mM) and low (Km = 7.6 mM) affinities for glucose. Chromatography of supernatants (105,000 X g) of insulinomas on DEAE-Cibacron Blue F3GA agarose evidenced glucose 6-phosphotransferase activity which eluted similarly to glucokinase from rat liver. Kinetic studies of insulinoma glucokinase also indicated similarity with liver glucokinase, i.e. cooperative rate dependence on glucose concentration and comparable Km values for glucose, and it did not phosphorylate N acetylglucosamine. These characteristics of glucose 6-phosphotransferase in insulinomas are similar to those of the enzyme found in islets of Langerhans. Since glucokinase is thought to serve as glucose sensor of insulin secretory pancreatic beta-cells, these transplantable insulinomas offer great promise for biochemical and biophysical studies of the nature of glucose-induced insulin release. PMID- 6296128 TI - Molecular size of opiate (enkephalin) receptors in neuroblastoma-glioma hybrid cells as determined by radiation inactivation analysis. AB - Opiate receptor binding decayed exponentially in mouse neuroblastoma-rat glioma (NG108-15) hybrid cell preparations following exposure to increasing doses of ionizing radiation (0.2 to 7.0 Mrads; 2.0 Mrads/min). Target size analysis revealed that [3H][D-Ala2, D-Leu5]enkephalin (agonist) and [3H]naloxone (antagonist) bound specifically to a component with an apparent molecular size of 200,000 +/- 20,000. Lyophilization of cells for the irradiation procedure did not significantly alter receptor affinity or binding capacity for these ligands. Furthermore, the loss of opiate receptor binding in irradiated cell samples could not be attributed to reduced receptor affinity since increasing concentrations of radiolabeled ligand failed to reverse the inhibition; nonspecific binding decreased only slightly under identical experimental conditions. The value of determining molecular size by radiation inactivation analysis was confirmed by showing that apparent target sizes for two representative lysosomal enzymes (beta galactosidase and alpha-mannosidase) were consistent with results obtained previously using conventional methods. Thus, the data suggest that the ligand binding component of delta-opiate (enkephalin) receptors in NG108-15 cells has a minimum functional size of approximately 200,000. PMID- 6296129 TI - Respiration-dependent proton translocation in alkalophilic Bacillus firmus RAB and its non-alkalophilic mutant derivative. AB - Obligately alkalophilic Bacillus firmus RAB had a higher molar growth yield on L malate (Ymal = 38 mg, dry weight/mmol of L-malate) than its non-alkalophilic mutant derivative, strain RABN (Ymal = 12 mg, dry weight/mmol of L-malate). Measurements of respiration dependent proton translocation by the two strains in the presence of K+ and valinomycin showed that the alkalophile also has much higher H+/O stoichiometries (at pH 9.0) than does the mutant (at pH 7.0). H+/O ratios for B. firmus RAB at pH 9.0 were as high as 13, with a frequently observed value of 9. These high values were observed in the first phase of a set of biphasic curves for both oxygen consumption and proton ejection. At pH 7.0, both the wild type and the mutant exhibited H+/O ratios near 4 in a single phase of oxygen consumption and proton ejection. The results are consistent with suggestions that the alkalophilic respiratory chain is especially well adapted for effective energy transduction at alkaline but not neutral pH. PMID- 6296130 TI - Semi-met oxidation level of chalcogenide derivatives of methemerythrin. Mossbauer and EPR studies. AB - Conclusive evidence is presented for an S = 1/2 spincoupled pair of high spin ferric and ferrous ions in the major reaction product of sulfide with the met form of the non-heme iron oxygen-carrying protein hemerythrin. Evidence for an analogous selenide derivative is also reported. Mossbauer and EPR spectroscopy establish (a) the charge and spin states of the individual iron atoms in sulfidehemerythrin as Fe(III), S = 5/2, and Fe(II), S = 2, and (b) the existence of an antiferromagnetic exchange interaction that couples the two spins to a resultant spin S = 1/2. The combined Mossbauer and EPR data confirm the correctness of the formulation first proposed for semi-methemerythrin by Harrington, P.C., de Waal, D.J.A., and Wilkins, R.G. ((1978) Arch. Biochem. Biophys. 191, 444-451) and furthermore show that a majority of the iron centers in the protein can be stabilized at this oxidation level. The results also demonstrate a new route to semi-methemerythrin. A titration of methemerythrin with selenide indicates that this derivative forms by a two step process consisting of first, reduction to the semi-met oxidation level by selenide and second, binding of selenide to either one or both irons. PMID- 6296131 TI - The oxidation of yeast Complex III. Evidence for a very rapid electron equilibration between cytochrome c1 and the iron-sulfur center. AB - The reoxidation of reduced cytochrome c1 by potassium ferricyanide follows pseudo first order kinetics with k = 4 x 10(4) M-1 s-1. However, the reoxidation of this cytochrome in two-electron reduced Complex III does not follow any simple rate law although the overall rate of reaction is essentially unchanged. The observed kinetics can be well fitted with a model in which ferricyanide reacts exclusively with cytochrome c1 together with very rapid electron transfer from the reduced iron-sulfur center to cytochrome c1. Neither removal of coenzyme Q from the complex nor prior incubation with antimycin A had any effect on the observed kinetics of reoxidation. PMID- 6296132 TI - Evidence that the Rous sarcoma virus transforming gene product is associated with glycerol kinase activity. AB - This communication provides biochemical, immunological, and genetic evidence that pp60src, the Rous sarcoma virus transforming gene product, is associated with glycerol kinase activity. Our investigations demonstrated that the compound phosphorylated by pp60src or by glycerol kinase (EC 2.7.1.30) from Candida mycoderma share the same electrophoretic and chromatographic mobilities. The glycerol kinase and protein kinase activities of pp60src were inhibited similarly by preincubation with immune IgG. Both activities were reduced 6-9-fold in pp60src preparations derived by immunoaffinity chromatography from cells which were infected with NY68, a temperature-sensitive transformation mutant of Rous sarcoma virus. The thermolability at 41 degrees C of the glycerol kinase activity of pp60src from the mutant virus-infected cells was greater (t/2 = 1.3 min) than the same activity in pp60src preparations from wild type virus-infected cells (t/2 = 4.8 min). PMID- 6296133 TI - Identification of phosphatidylinositol kinase in rat liver lysosomal membranes. AB - Liver lysosomes from Triton-injected or normal rats were found to rapidly incorporate 32P from [gamma-32P]ATP into a lipid component of the membrane, in vitro. The lipid was identified as phosphatidylinositol 4-phosphate based on its chromatographic behavior on Silica Gel H thin layer plates as compared with standard phosphoinositides. The deacylation product, glyceryl-phosphorylinositol phosphate, was compared with standards in chromatographic and electrophoretic systems to further substantiate the identification of the radioactive material. A trace of phosphatidylinositol 4,5-bisphosphate was also found. The properties of the lysosomal membrane phosphatidylinositol kinase were examined using both endogenous lipid and exogenous phosphatidylinositol as substrate. The enzyme was active at neutral pH in the presence of 20 mM MgCl2. The addition of 0.4% Triton X-100 stimulated the enzyme activity toward endogenous substrate, and the highest activity was observed in the presence of detergent and 1 mM phosphatidylinositol. Degradation of the product was seen only in the presence of Triton X-100. The specific activity of the lysosomal phosphatidylinositol kinase is comparable to the detergent-stimulated activity of liver microsomes and plasma membrane, the previously recognized sources of this enzyme in the liver cell. PMID- 6296134 TI - Identification of endorphin acetyltransferase in rat brain and pituitary gland. AB - An enzyme which N-acetylates beta-endorphin has been identified and characterized in the rat brain and pituitary gland. The beta-endorphin acetyltransferase activity was localized almost exclusively in the intermediate lobe of the pituitary gland. beta-endorphin acetyltransferase activity was also present in the hypothalamus, a brain region which synthesizes beta-endorphin, but not the cerebellum, a region which does not synthesize beta-endorphin. The substrate specificity of beta-endorphin acetyltransferase appeared to be located in the NH2 terminus and midportions of the beta-endorphin sequence. The regional distribution and pharmacological specificity of beta-endorphin acetyltransferase indicate that it is a highly specialized regulatory enzyme. Because N-acetyl-beta endorphin is nonanalgesic and does not bind to the opiate receptor (Smyth, D.G., Massey, D.E., Zakarian, S., and Finnie, M.D. (1979) Nature (Lond.) 279, 252-254), it appears that the physiological significance of beta-endorphin acetyltransferase is that it can modulate the activity of endorphin secreted from opiomelanotropinergic cells and neurons. PMID- 6296135 TI - Raman difference spectroscopy of heme-linked ionizations in cytochrome c peroxidase. AB - The pH dependence of the oxidation-state marker line of hemoproteins is investigated in cytochrome c peroxidase with Raman difference spectroscopy. The frequency is sensitive to ionization of a group on the protein that regulates catalytic activity of the resting ferriheme enzyme. The oxidation-state marker line shows a transition with pK of 5.5 in good agreement with other spectroscopic measurements and kinetic measurements of binding of peroxide, and other ligands to the native enzyme. The shift of 0.8 cm-1 to higher frequency at pH 4.5 relative to the pH 6.4 value is interpreted in terms of a substantial decrease in pi-electron density in the porphyrin ring. Charge density in the pi-system is highest at maximal activity, as would be expected if donor-acceptor interactions with residues of the protein stabilize the oxidized Fe(IV) reaction intermediate. Evidence of additional heme-linked ionizations with pK values near 7.5 is found; this alkaline transition involves deprotonation of several groups of the protein, conversion of iron from high to low spin, and, possibly, denaturation of the protein. PMID- 6296136 TI - A spin label substrate analogue as active site-directed modifying agent. Tryptophan 140 of aspartate aminotransferase. PMID- 6296137 TI - Disruption of erythrocyte membranal organization by superoxide. AB - Human erythrocyte ghosts were covalently labeled with 4-maleimide-2,2,6,6 tetramethylpiperidinooxyl. Electron paramagnetic resonance (EPR) spectrometry revealed two major binding environments representing strongly (S) and weakly (W) immobilized species. The disorder parameter, W/S, determined from the respective peak amplitudes, was shown to be irreversibly elevated following treatment of the labeled ghosts with superoxide, indicating an increase in membrane fluidity. Labeled ghosts reduced with ascorbate showed no nitroxide EPR signals. However, following exposure of these membranes to superoxide, the nitroxide spectrum returned with a W/S ratio of 25. In contrast, the disorder parameter for spin labeled ghosts decreased following exposure to hydroxyl radicals suggesting decreased fluidity, as a result of lipid peroxidation. This effect could be prevented by the inclusion of mannitol. These changes in membrane fluidity and/or protein mobility observed by EPR are compared with previous results obtained by other methods and provide additional evidence for physiologic alterations initiated by superoxide. PMID- 6296138 TI - GDP does not mediate but rather inhibits hormonal signal to adenylate cyclase. AB - This study was aimed to elucidate whether GDP can mediate hormonal signal to adenylate cyclase in hepatic glucagon sensitive adenylate cyclase with ATP as substrate. Conversion of added GDP to GTP catalyzed by nucleoside diphosphate kinase was suppressed to less than 0.3% of added GDP by including UDP. Inhibition of this enzyme activity by UDP was accompanied by a preferential loss of the stimulatory effect of glucagon plus GDP on cyclase activity without changes in effects of glucagon plus GTP, glucagon plus guanosine 5'-(beta, gamma imino)triphosphate, and NaF. Under this condition, i.e. in the presence of UDP, GDP competitively inhibited the actions of GTP (Ki for GDP, 1 microM) and guanosine 5'-(beta, gamma-imino)triphosphate in the presence of glucagon, the inhibition being complete at high GDP concentrations. GDP also inhibited cyclase activity stimulated by NaF with UDP but did only slightly without UDP. It was demonstrated that nucleoside diphosphate kinase is located in membranes in addition to cytosol fraction. However, the activity of membrane-associated enzyme was not affected by the addition of glucagon. Based on these observations, it is concluded that GDP is unable to mediate hormonal signal to adenylate cyclase and that it acts as an inhibitor of cyclase activity stimulated by GTP or its analog along with hormone. The results suggest a possible role of membrane-associated nucleoside diphosphate kinase in determining GTP and GDP levels at or near their binding site so as to replenish GTP and, thereby, decrease the inhibitory action of GDP when hormone is present. PMID- 6296139 TI - Purification and properties of Pseudomonas aeruginosa porin. PMID- 6296140 TI - Specific mixed disulfide formation with purified bovine cardiac glycogen synthase I and glutathione. AB - Bovine cardiac glycogen-free glycogen synthase I reacts with oxidized glutathione at low temperature to partially inactivate the enzyme. Evidence is presented that a mixed disulfide between glutathione and the enzyme is formed in this reaction. A short incubation of the GSSG-treated enzyme with dithiothreitol restores full enzyme activity. The reaction with GSSG is pH dependent and the product is quite stable at neutral pH. Oxidation of one sulfhydryl group in glycogen synthase is associated with a loss of 60-70% of the enzyme activity. Further modification of protein sulfhydryls has less effect on the enzyme activity. Other low molecular weight disulfides also inactivate glycogen synthase and treatment with [35S]cystine to produce a 40% loss of enzyme activity gave rise to a single major radioactive peptide after cyanogen bromide digestion. Thus the GSSG-mediated inactivation of glycogen synthase apparently occurs through a single reactive sulfhydryl group that forms a mixed disulfide with low molecular weight disulfide molecules. Uridine 5'-diphosphate glucose and glycogen prevent the inactivation of glycogen-free glycogen synthase with GSSG, and glucose 6-phosphate retards the rate of inactivation. Reduction and reactivation of the GSSG-oxidized glycogen synthase is not affected by glycogen and it occurs readily at neutral pH with dithiothreitol, mercaptoethanol, or cysteamine. Oxidation of the reactive sulfhydryl group with GSSG has no effect on the rate of glycogen synthase phosphorylation by the catalytic subunit of cAMP-dependent protein kinase. PMID- 6296141 TI - Agonist and antagonist affinities for inhibitory adenosine receptors are reciprocally affected by 5'-guanylylimidodiphosphate or N-ethylmaleimide. PMID- 6296142 TI - Cloning and nucleotide sequence analysis of the dog insulin gene. Coded amino acid sequence of canine preproinsulin predicts an additional C-peptide fragment. AB - A 4.0-kilobase HindIII/EcoRI-cleaved dog genomic DNA fragment was shown to contain the dog insulin gene by restriction mapping using a human insulin cDNA probe. This fragment was subsequently cloned in a lambda vector, and the nucleotide sequence of the dog insulin gene was determined. As in several other species, the insulin gene of the dog is interrupted by two intervening sequences, one of 151 base pairs located in the 5' untranslated region and the other of 264 base pairs occurring within the codon of the 7th amino acid of the C-peptide. Translation of the nucleotide sequence in one frame revealed the primary structure of canine preproinsulin. An interesting feature of the coded amino acid sequence is that it predicts a C-peptide of 31 amino acids, 8 residues longer than that reported by Peterson et al. (Peterson, J. D., Nehrlich, S., Oyer, P. E., and Steiner, D. F. (1973) J. Biol. Chem. 247, 4866-4871). The additional octapeptide sequence, Glu-Val-Glu-Asp-Leu-Gln-Val-Arg, is located NH2-terminal to the 23-residue C-peptide sequence described in the earlier report. Its coding sequence is interrupted by the second intervening sequence. The arginine at position 8 suggests that a trypsin-like cleavage may separate the NH2-terminal octapeptide from the remainder of the C-peptide during the post-translational processing of dog proinsulin in the pancreas. The revised C-peptide sequence suggests that the proinsulin C-peptide is more highly conserved in length and overall sequence than was previously supposed. PMID- 6296143 TI - Purification and characterization of rabbit liver calmodulin-dependent glycogen synthase kinase. AB - A rabbit liver cAMP-independent glycogen synthase kinase has been purified 4500 fold to a specific activity of 2.23 mumol of 32P incorporated per min per mg of protein using ion exchange chromatography on DEAE-Sephacel and phosphocellulose, gel filtration chromatography on Sepharose 6B, and affinity chromatography on calmodulin-Sepharose. This synthase kinase, which was completely dependent on the presence of calmodulin (apparent K0.5 = 0.1 microM) and calcium for activity, also catalyzed the phosphorylation of purified smooth muscle myosin light chain but not of smooth muscle myosin. Using 0.5 mM ATP, a maximal rate of phosphorylation of glycogen synthase was achieved in the presence of 10 mM magnesium acetate with a pH optimum of 7.8. Gel filtration experiments indicated a Stokes radius of about 70 A and sucrose density gradient centrifugation data gave a sedimentation coefficient of 10.6 S. A molecular weight of approximately 300,000 was calculated. A definitive subunit structure was not determined, but major bands observed after polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate corresponded to a doublet at 50,000 to 53,000. The calmodulin-dependent glycogen synthase kinase incorporated about 1 mol of 32P per mol of synthase subunit into sites 2 and 1b associated with a decrease in the synthase activity ratio from 0.8 to about 0.4. The calmodulin-dependent glycogen synthase kinase may mediate the effects of alpha-adrenergic agonists, vasopressin, and/or angiotensin II on glycogen synthase in liver. PMID- 6296144 TI - The biosynthesis of membrane-derived oligosaccharides. A membrane-bound phosphoglycerol transferase. AB - Membrane-derived oligosaccharides, found in the Escherichia coli periplasmic space (Schulman, H., and Kennedy, E. P. (1979) J. Bacteriol. 137, 686-688), are composed of 8-10 units of glucose, the sole sugar, in beta 1 leads to 2 and beta 1 leads to 6 linkages (Schneider, J. E., Reinhold, V., Rumley, M. K., and Kennedy, E. P. (1979) J. Biol. Chem. 254, 10135-10138). Oligosaccharides in this family are variously substituted with succinyl ester residues, as well as with sn 1-phosphoglycerol and phosphoethanolamine, both derived from membrane phospholipids. These negatively charged oligosaccharides may function in cellular osmoregulation since their synthesis is under osmotic control (Kennedy, E. P. (1982) Proc. Natl. Acad. Sci. U.S.A. 79, 1092-1095). We now report initial characterization of an enzyme catalyzing transfer of phosphoglycerol residues from phosphatidylglycerol to membrane-derived oligosaccharides or to synthetic beta-glucoside acceptors. The products are sn-1,2-diglyceride and beta-glucoside 6-phosphoglycerol. Localized in the inner membrane, the transferase has a requirement for divalent cations, of which manganese is most effective, and a pH optimum of 8.9 in vitro. PMID- 6296145 TI - Ion-gated channel induced in planar bilayers by incorporation of (Na+,K+)-ATPase. AB - An ion-gated channel was conferred on a planar lipid bilayer membrane upon incorporation of (Na+,K+)-ATPase. The channel exhibited two conductance states. The high conductance state was only observed when an ion gradient was present across the planar membrane. This state corresponded to an enzyme conformation which was ouabain and vanadate sensitive (i.e. conductance was inhibited by these compounds), while the low conductance state showed no sensitivity to either inhibitor. Single channel conductance behavior was observed when minimal amounts of enzyme were incorporated into the planar bilayer. The observed single channel conductance was 270 +/- 14 picosiemens. Similar transport behavior was observed for enzyme purified from ovine kidney using sodium dodecyl sulfate (anionic), eel electroplax using Lubrol-WX (nonionic), and kidney microsomes. In addition, the data strongly suggest that enzyme from the kidney microsomes was asymmetrically incorporated into the planar bilayer. PMID- 6296146 TI - Calmodulin interacts with cyclic nucleotide phosphodiesterase and calcineurin by binding to a metal ion-independent hydrophobic region on these proteins. AB - Hydrophobic interaction chromatography is employed to determine if calmodulin might associate with its target enzymes such as cyclic nucleotide phosphodiesterase and calcineurin through its Ca2+-induced hydrophobic binding region. The majority of protein in a bovine brain extract that binds to a calmodulin-Sepharose affinity column also is observed to bind in a metal ion independent manner to phenyl-Sepharose through hydrophobic interactions. Cyclic nucleotide phosphodiesterase activity that is bound to phenyl-Sepharose can be resolved into two activity peaks; one peak of activity is eluted with low ionic strength buffer, while the second peak eluted with an ethylene glycol gradient. Calcineurin bound tightly to the phenyl-Sepharose column and could only be eluted with 8 M urea. Increasing ethylene glycol concentrations in the reaction mixture selectively inhibited the ability of calmodulin to stimulate phosphodiesterase activity, suggesting that hydrophobic interaction is required for activation. Comparison of the proteins which are bound to and eluted from phenyl- and calmodulin-Sepharose affinity columns indicates that chromatography involving calmodulin-Sepharose resembles hydrophobic interaction chromatography with charged ligands. In this type of interaction, hydrophobic binding either is reinforced by electrostatic attractions or opposed by electrostatic repulsions to create a degree of specificity in the binding of calmodulin to certain proteins with accessible hydrophobic regions. PMID- 6296147 TI - Properties of the adenylate cyclase catalytic unit from caudate nucleus. PMID- 6296148 TI - Tetrodotoxin-sensitive sodium channels in rat muscle cells developing in vitro. PMID- 6296149 TI - Phosphorylation of renal brush-border membrane vesicles. Effect on calcium uptake and membrane content of polyphosphoinositides. PMID- 6296150 TI - Two-dimensional arrays of proteins in sarcoplasmic reticulum and purified Ca2+ ATPase vesicles treated with vanadate. PMID- 6296151 TI - Peptide mapping of the human transferrin receptor in normal and transformed cells. AB - Human transferrin receptor protein from various human cell lines in tissue culture (phytohemagglutinin-activated lymphocytes, normal embryonic fibroblasts (WI-38), a choriocarcinoma (Be Wo), an epidermoid carcinoma (KB), and a metastatic breast carcinoma (MCF-7)) was metabolically labeled with [35S]methionine and purified by two methods. The first method involves affinity chromatography of the transferrin receptor with a transferrin-linked Sepharose 4B resin. The second method involves direct immunoprecipitation of the receptor protein with a goat polyclonal antibody raised against placental transferrin receptor. Additionally, transferrin receptor from normal gestational placenta was similarly radiolabeled with [35S]methionine and subsequently purified with transferrin-bound resin. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the reduced receptor isolated by both purification techniques revealed an Mr = 94,000 protein to be present in all cells assayed for this study. Partial proteolytic digestion with Staphylococcus aureus protease followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed identical mapping patterns of the Mr = 94,000 protein isolated from each cell line, irrespective of the isolation technique employed to purify the receptor. These findings indicate an identical peptide structure of the human transferrin receptor in cells with a differentiated function for iron metabolism, in normal cells undergoing mitosis, and in neoplastic cells in long term culture, within the limits of detectability of the proteolytic digestion maps. PMID- 6296153 TI - Covalent coupling of human growth hormone to its receptor on rat hepatocytes. PMID- 6296152 TI - Mechanism of the rifampicin induction of RNA polymerase beta and beta' subunit synthesis in Escherichia coli. AB - The mechanism of the rifampicin induction of RNA polymerase beta and beta' subunit synthesis was investigated, employing an in vitro coupled system of transcription and translation as well as an in vitro transcription system with purified RNA polymerase holoenzyme. Two independent effects leading to the induction of beta and beta' polypeptide synthesis have been identified for the drug: 1) rifampicin binds to RNA polymerase holoenzyme and inhibits its autorepressor activity, thus relieving the autorepression exerted possibly on the translation of rpoBC mRNA; and 2) rifampicin-RNA polymerase complex functions as a positive effector stimulating the transcription of rpoBC genes, but not of rplL gene. Using a terminally labeled DNA fragment covering the intercistronic region between rplL and rpoB as a probe, the structure of both in vivo and in vitro RNA were analyzed by S1 nuclease-mapping assays. The experimental results indicated that the observed stimulation of the transcription by rifampicin was attributed, at least in part, to relaxation of the transcription attenuation immediately preceding the rpoB gene, resulting in the increase of the read-through transcript to rpoBC genes. Besides the read-through transcript and the processed mature rpoBC mRNA, another RNA species was found in in vivo RNA, which might be transcribed from a weak promoter located in the intercistronic region. However, no clear indication has been obtained of possible enhancement of the transcription initiation by rifampicin from the putative promoter. PMID- 6296154 TI - Estimation of the bacterial content of pertussis vaccines by dry weight and nitrogen determinations of bacterial mass. PMID- 6296155 TI - 125I-thrombin binds to clustered receptors on noncoated regions of mouse embryo cell surfaces. AB - We used electron microscope autoradiography (EMAR) to visualize the interaction of 125I-thrombin with its surface receptors on mouse embryo (ME) cells. Autoradiographic grains were spaced over the surface of cells in a periodic nonrandom pattern, indicating 125I-thrombin association with clusters of thrombin receptors. The grain spacing varied slightly from cell to cell, indicating subpopulations of cells with different numbers of thrombin receptors. The average distance between grains on ME cells after binding 125I-thrombin (125 ng/ml) at 37 degrees C was 1.65 +/- 0.49 microns. The average distance between grains on prefixed cells and cells incubated with 125I-thrombin at 4 degrees C was not significantly different from that observed at 37 degrees C. This indicates that thrombin receptors are clustered before thrombin binding and that the thrombin receptor aggregates do not redistribute into large aggregates on the surface of cells subsequent to thrombin binding. The number of grains per cluster also does not change under these three binding conditions. Thus, the number of occupied receptors in each cluster appears to be constant. On the basis of the average grain number and spacing, we estimate that each cluster is approximately 400 nm in diameter containing approximately 550 thrombin-binding sites. These receptor clusters are not associated with specialized structures or coated regions of the membrane. Additionally, grains observed within cells were not found associated with coated vesicles. Therefore, neither the clustering patterns nor internalization of 125I-thrombin are characteristic of molecules which bind to receptors and are internalized by receptor-mediated endocytosis. PMID- 6296156 TI - Development of cytochrome b and an active oxidase system in association with maturation of a human promyelocytic (HL-60) cell line. AB - The human HL-60 myeloid leukaemia cell line developed, during maturational changes induced by dimethyl sulphoxide, an enhanced capacity for phorbol myristate acetate- stimulated oxidative activity and acquired a cytochrome b. Titration of the absorbance at 559 nm at potentials of-190 to -370 mV indicated that this cytochrome had a very low potential, differentiating it from mitochondrial and endoplasmic reticulum cytochromes and identifying it as the cytochrome b(-245) that has been recently found in other phagocytic cells. Subcellular fractionation studies of mature HL-60 cells showed that cytochrome b had a dual distribution within the cell. The lighter peak of activity was associated with the plasma membrane markers, adenylate cyclase and receptors for the N- formal-L-methionyl-L-leucyl-L-phenylalanine (f-Met-Leu-Phe) peptide. The denser components localized with the mitochondria but were distinct from mitochondrial cytochromes because whereas the activity of cytochrome c oxidase fell during HL-60 cell maturation, that of this cytochrome b was markedly increased. Concentrations of myeloperoxidase were unrelated to activity of the oxidase system and decreased as the cell matured. The increase in the concentrations of cytochrome b with cellular maturation parallelled the increase in the stimulated nonmitochondrial respiratory activity of these cells. The turnover of the hexose monophosphate shunt of immature cells was increased by the oxidising agents, methylene blue and tert-butylhydroperoxide, indicating that these immature cells have stimulated nonmitochondrial respiratory activity by maturing HL-60 cells is associated with, and is probably dependent upon, the acquisition by these cells of the cytochrome b(-245) oxidase system. PMID- 6296157 TI - Diffusion of low density lipoprotein-receptor complex on human fibroblasts. AB - Diffusion of the complex consisting of low density lipoprotein (LDL) bound to its receptor on the surface of human fibroblasts has been measured with the help of an intensely fluorescent, biologically active LDL derivative, dioctadecylindocarbocyanine LDL (dil(3)-LDL). Fluorescence photobleaching recovering and direct video observations of the Brownian motion of individual LDL receptor complexes yielded diffusion coefficients for the slow diffusion on cell surfaces and fast diffusion on membrane blebs, respectively. At 10 degrees C, less that 20 percent of the LDL-receptor complex was measurably diffusible either on normal human fibroblasts GM-3348 or on LDL-receptor- internalization-defective J.D. cells GM-2408A. At 21 degrees and 28 degrees C, the diffusion fractions of approximately 75 and 60 percent, respectively, on both cell lines. The lipid analog nitrobenzoxadiazolephosphatidylcholine (NBD-PC) diffused in the GM-2408A cell membrane at 1.5x10(-8) cm(2)/sec at 22 degrees C. On blebs induced in GM 2408A cell membranes, the dil(3)-LDL receptor complex diffusion coefficient increased to approximately 10(-9) cm(2)/s, thus approaching the maximum theoretical predictions for a large protein in the viscous lipid bilayer. Cytoskeletal staining of blebs with NBD- phallacidin, a fluorescent probe specific for F-actin, indicated that loss of the bulk of the F-actin cytoskeleton accompanied the release of the natural constraints on later diffusion observed on blebs. This work shows that the internalization defect of J.D. is not due to immobilization of the LDL-receptor complex since its diffusibility is sufficient to sustain even the internalization rates observed in the native fibroblasts. Nevertheless, as with many other cell membrane receptors, the diffusion coefficient of the LDL-receptor complex is at least two orders of magnitude slower on native membrane than the viscous limit approached on cell membrane blebs where it is released from lateral constraints. However, LDL-receptor diffusion may not limit LDL internalization in normal human fibroblasts. PMID- 6296158 TI - Distribution of an asialoglycoprotein receptor on rat hepatocyte cell surface. AB - Direct ferritin immunoelectron microscopy was applied to visualize the distribution of the hepatocyte cell surface of the asialoglycoprotein receptor which is responsible for the rapid clearance of serum glycoproteins and lysosomal catabolism. For this purpose, rabbit antibody against the purified hepatic binding protein specific for asialoglycoproteins was prepared and coupled to ferritin by glutaraldehyde. The specific antibody conjugates were incubated with the hepatocytes, which were isolated from rat liver homogenate after fixation by glutaraldehyde perfusion. These cells preserved well the original polygonal shape and polarity, and it was easy to identify the sinusoidal, lateral, and bile canalicular faces. The surface density of the ferritin particles bound to the sinusoidal face was about four times higher than that of particles bound to the lateral face, while the bile canalicular face was hardly labeled and almost at the control level. Using the surface area of hepatocyte measured by morphometrical analyses, it was estimated that approximately 90% of bound ferritin particles were at the sinusoidal face, approximately 10% at the lateral face, and approximately 1% at the bile canalicular face. Nonhepatic cells such as endothelial and Kupffer cells had no receptor specific for asialoglycoproteins. PMID- 6296159 TI - Inhibition of fibronectin receptor function by antibodies against baby hamster kidney cell wheat germ agglutinin receptors. AB - Previous studies suggest that the baby hamster kidney (BHK) cell fibronectin receptor is also a wheat germ agglutinin receptor (WGA-R). To analyze this possibility further, IgG and Fab fragments of antibodies produced against a BHK cell WGA-R preparation were tested to determine their effects on cell adhesion mediated by fibronectin, wheat germ agglutinin, concanavalin A, and polycationic ferritin. The WGA-R preparation was isolated by octylglucoside extraction of BHK cells followed by chromatography of the extract on WGA-agarose. The antibodies against the WGA-R preparation reacted primarily with polypeptides of molecular weights 48, 61, 83, 105, 120, 165, 210, and 230 kilodaltons (kdaltons). It was concluded that the antibodies interfered with BHK cell fibronectin receptors on the basis of the ability of anti-WGA-R IgG or Fab fragments to (a) inhibit cell spreading on fibronectin-coated substrata; (b) cause rounding and detachment of cells previously spread on fibronectin-coated substrata; and (c) inhibit binding of fibronectin-coated latex beads to the cells. Antibody activity was blocked by treatment of anti-WGA-R with the WGA-R preparation or by absorption of anti-WGA-R with intact BHK cells. The antibodies also appeared to prevent coupling of ligand receptor complexes (involving concanavalin A or polycationic ferritin) with the cytoskeleton. Finally, cell rounding and detachment caused by the antibodies were found to require metabolic energy since it did not occur in the presence of azide or at 4 degrees C. PMID- 6296160 TI - Hormone-induced protein phosphorylation. I. Relationship between secretagogue action and endogenous protein phosphorylation in intact cells from the exocrine pancreas and parotid. AB - We undertook studies to determine whether secretagogue action on the exocrine pancreas and parotid is accompanied by phosphorylation of proteins in intact cells. For this purpose, rat pancreatic, and parotid lobules were preincubated with 32Pi for 45 min at 37 degrees C, washed, and then incubated at 37 degrees C in the presence or absence of secretagogues that effect discharge through different second messengers. Among a variety of polypeptides exhibiting enhanced phosphorylation in pancreatic lobules upon a 30-s incubation in the presence of the secretagogues carbamylcholine, cholecystokinin octapeptide, or secretin, one species with an Mr of 29,000 was especially notable for three reasons: (a) its enhanced level of phosphorylation was dependent on the dose of secretagogue used and was still apparent after incubation for 30 min at 37 degrees C; (b) an analogous phosphorylated polypeptide was observed in isoproterenol-stimulated parotid lobules; and (c) in both tissues its selective dephosphorylation was observed upon termination of stimulation by administration of atropine to carbamylcholine-stimulated pancreatic lobules and propranolol to isoproterenol stimulated parotid lobules. These results suggest that the phosphorylation of one protein with an Mr of 29,000 is closely correlated both temporally and in a dose dependent fashion with secretagogue action in both the exocrine pancreas and parotid. PMID- 6296161 TI - Hormone-induced protein phosphorylation. II. Localization to the ribosomal fraction from rat exocrine pancreas and parotid of a 29,000-dalton protein phosphorylated in situ in response to secretagogues. AB - In the preceding paper, we demonstrated that the endogenous phosphorylation of a protein with a molecular weight of 29,000 was enhanced by various secretagogues in rat pancreatic and parotid lobules, the phosphorylation of this protein correlating both temporally and in a dose-dependent fashion with secretory protein discharge. In the present study, we established a specific methodology to characterize this phosphoprotein. Once established, this 29,000-dalton phosphoprotein was then followed selectively and quantitatively throughout subcellular fractionation procedures. Analysis of two-dimensional polyacrylamide gels demonstrated that proteins with similar mobilities (Mr 29,000; pl greater than 8.4) were affected by cholecystokinin octapeptide and isoproterenol in rat pancreatic and parotid lobules, respectively, suggesting that the same 29,000 dalton phosphoprotein was covalently modified in both tissues. Cellular fractionation studies using differential velocity and sucrose density gradient centrifugation revealed that the 29,000-dalton phosphoprotein copurified with the rough microsomal fraction of pancreas and was highly enriched in ribosomal fractions of both pancreas and parotid. Electrophoresis in two dimensions confirmed that the 29,000-dalton polypeptide that was resolved directly from stimulated cells and from ribosomal fractions exhibited a common mobility, and apparent identity of the species was strongly suggested when the 29,000-dalton polypeptides from both sources were compared by peptide mapping following limited digestion with Staphylococcus aureus V8 protease. This phosphoprotein was tentatively identified as ribosomal protein S6 after analysis by pH 8.6/4.2 two dimensional PAGE. PMID- 6296162 TI - Hormone-induced protein phosphorylation. III. regulation of the phosphorylation of the secretagogue-responsive 29,000-dalton protein by both Ca2+ and cAMP in vitro. AB - In the preceding papers, we demonstrated that the endogenous phosphorylation of a 29,000-dalton protein is stimulated in response to secretagogue application to intact cells from the rat exocrine pancreas and parotid and dephosphorylated upon termination of secretagogue action. One- and two-dimensional gel analysis of 32Pi labeled pancreatic and parotid lobules as well as their respective subcellular fractions revealed that the same protein was covalently modified in both tissues and was localized to the ribosomal fraction. To identify the intracellular second messengers which may mediate or modulate the phosphorylation of the 29,000-dalton protein in intact cells, the effects of Ca2+, cAMP, and cGMP on the endogenous phosphorylation of this protein were assessed in subcellular fractions from the rat pancreas and parotid. Our results demonstrate that the phosphorylation of the 29,000-dalton polypeptide may be regulated by both Ca2+ and cAMP in the pancreas and in the parotid. No cGMP-dependent protein phosphorylation was found in either tissue. As in the in situ phosphorylation studies, the Ca2+- and cAMP-dependent phosphorylation of this same protein was localized to the ribosomal fraction. The cAMP-dependent protein kinase activity was found primarily in the postmicrosomal supernatant in contrast to the Ca2+-dependent protein kinase that appeared to be tightly associated with the substrate in addition to being present in the postmicrosomal supernatant. The data suggest that, in cells from the exocrine pancreas and parotid, secretagogues may regulate the phosphorylation of the 29,000-dalton protein through Ca2+ and/or cAMP. PMID- 6296163 TI - Glycerol-3-phosphate dehydrogenase is induced by glucocorticoids in hepatocytes and hepatoma cells in vitro. AB - Previous studies have shown that cytosolic glycerol-3-phosphate dehydrogenase (GPDH; EC 1.1.1.8) can be induced by glucocorticoids in mammalian brain, mammary gland, and thymus, but it was thought that no induction occurred in liver. We report here that GPDH is induced by glucocorticoids in several lines of hepatoma cells and in rat hepatocytes cultured in vitro. When rat hepatoma cells of clone FU5AH were exposed to 3 microM hydrocortisone (HC) for 3 days, GPDH specific activity increased greater than sixfold over control. The rate and extent of induction were similar in exponentially growing and stationary-phase cultures of cells. Four other hepatoma cell lines were inducible to a lesser extent, and three lines were not inducible. GPDH was also induced by glucocorticoids in cultures of hepatocytes isolated from livers of 6-day-old rats. The enzyme was induced three-to fourfold by the synthetic glucocorticoid, dexamethasone, in the presence of 1 nM insulin, but the induction was not observed in the absence of insulin. PMID- 6296164 TI - Receptor-mediated endocytosis and exocytosis of transferrin in Concanavalin A stimulated rat lymphoblasts. AB - Iron-loaded transferrin has been shown to be necessary for the support of cell proliferation in culture. This function depends upon interaction of transferrin with a specific high-affinity cell surface receptor. The present report is directed toward determining the consequences of the interaction of transferrin with this receptor on Concanavalin A-stimulated rat lymphocytes. Three specific questions have been posed: a) Is transferrin endocytosed following binding to its specific receptor in a temperature-dependent fashion? b) Following endocytosis, is the carrier protein released from the cell in a structurally and functionally intact form? and c) Is the cell surface transferrin receptor also endocytosed following ligand binding? The results provide affirmative answers to all questions. Using two independent probes of the cell surface versus intracellular location of transferrin we observed that cell-bound transferrin moved from the cell surface to the inside of the cell and subsequently back to the medium. This process occurred in a temperature-dependent fashion. When cells containing only intracellular transferrin were further incubated at 37 degrees C approximately 80% of cell-bound transferrin was released to the medium. Nearly all of this material retained reactivity with antibody to transferrin. In addition, endocytosed transferrin exhibited qualitatively and quantitatively equivalent binding reactivity with the transferrin receptor and showed identical electrophoretic mobility on SDS gel electrophoresis. Finally, using similar methodology to that employed with transferrin itself, we provide evidence that the specific receptor is also endocytosed. PMID- 6296165 TI - Ionic behaviors and neuronal survival in developing ganglia. III. Studies with embryonic chick sympathetic neurons. AB - We have shown in the past that (1) Nerve Growth Factor (NGF) controls the Na+, K+ pump in its ganglionic neuronal targets and (2) the NGF requirement for pump control is developmentally regulated in the chick embryo dorsal root ganglion. We report here that NGF is fully competent to insure the control of intracellular Na+ concentrations (as expression of pump control) in intact chick sympathetic ganglia and enriched suspensions of sympathetic neurons from embryonic day 8 (E8) through 13. At later stages (E13-E18), NGF becomes less and less required for that control as the neurons gain a self-sustained ionic pump competence. In monolayer cultures of enriched sympathetic neurons, an increasing neuronal survival in the absence of NGF occurs. These data demonstrate that the ability of developing sympathetic neurons to survive without NGF increases with the same temporal pattern as does their independence from NGF for ionic pump control, stressing the importance of ionic events for neuronal survival. PMID- 6296166 TI - Modulation of epidermal growth factor-dependent protein phosphorylation in cell membrane preparations by receptor down regulation. PMID- 6296167 TI - Modulation of the neuronal binding of the beta subunit of nerve growth factor (NGF) by the alpha-NGF subunit. AB - The effect of the alpha subunit of the 7S-NGF on the binding of beta-NGF to its two classes of sites on target cells has been studied. The presence of microM concentrations of alpha-NGF causes the displacement of 125I-beta-NGF from one class of sites on dissociated dorsal root ganglia neurons from stage E9 chicken embryos. At 0.1 nM 125I-beta-NGF, increasing alpha-NGF concentrations produce a monotonic displacement curve with half-maximal displacement occurring at 10 microM alpha-NGF. The affinity and number of sites of the 125I-beta-NGF displaced by alpha-NGF are similar to those of beta-NGF that binds to the higher affinity (site I) receptors. The binding to the lower affinity class of sites (site II) is not affected by concentrations of alpha-NGF up to 30 microM. This modulation of 125I-beta-NGF binding does not occur with equivalent concentrations of serum albumin. No detectable neuronal binding of 125I-alpha-NGF was found, suggesting that the mechanism does not involve direct competition for receptor sites. The dissociation constant for the alpha-beta complex is in the microM range, and formation of this complex in solution can thus compete with the process of 125I beta-NGF binding to neurons. A model accounting for these observations includes binding of the alpha-beta complex to the lower affinity but not to the higher affinity sites. We conclude that there are differences in the specificity of the two classes of receptors. PMID- 6296168 TI - Multiple specific binding sites for purified glucocorticoid receptors on mammary tumor virus DNA. AB - Glucocorticoid hormones selectively stimulate the rate of transcription of integrated mammary tumor virus (MTV) sequences in infected rat hepatoma cells. Using two independent assays, we find that purified rat liver glucocorticoid receptor protein binds specifically to at least four widely separated regions on pure MTV proviral DNA. One of these specific binding domains, which itself contains at least two distinct receptor binding sites, resides within a fragment of viral DNA that maps 110-449 bp upstream of the promoter for MTV RNA synthesis. Three other binding domains lie downstream of the promoter and within the MTV primary transcription unit. Restriction fragments bearing separate binding domains have been introduced into cultured cells; transformants have been recovered in which the introduced fragments are expressed under glucocorticoid control. Thus, it appears that this assay will be useful for assessing the biological significance of the receptor binding sites detected in vitro. PMID- 6296169 TI - Role of epidermal growth factor-stimulated protein kinase in control of proliferation of A431 cells. AB - Epidermal growth factor (EGF), which stimulates tyrosine-specific protein kinase activity both in vivo and in vitro, inhibits proliferation of A431 human epidermoid carcinoma cells. After mutagenesis clonal cell lines that were resistant to the growth inhibitory effects of EGF were selected. All six variants examined contained decreased EGF-stimulated protein kinase. The number of EGF receptors in variant cells decreased in parallel with EGF-stimulated protein kinase activity so that the specific activity of EGF-stimulated protein kinase per EGF receptor remained constant in variant cell lines with up to tenfold reductions in both activities. This result suggests that both EGF binding and kinase activities reside in the same or closely coupled molecules. The effect of EGF on growth of two resistant variants was examined in detail. Clone 29 contains approximately 50% and clone 4 contains approximately 20% of the EGF-stimulated protein kinase activity of the parental A431 cell line. In serum-supplemented medium, EGF stimulated proliferation of clone 29 but did not affect growth of clone 4. In a 1:1 mixture of DME and F-12 medium without serum, EGF caused both clone 29 and clone 4 to grow as well as in 10% serum. These variants, which were selected for resistance to the growth inhibitory effects of EGF, thus exhibit a strong mitogenic response to EGF. This result suggests that resistance to the growth inhibitory effect of EGF may involve both a decrease in EGF-stimulated protein kinase and an alteration in the response pathway. PMID- 6296170 TI - Sequence analysis and in vitro transcription of portions of the Epstein-Barr virus genome. AB - The 17,180 base-pair Eco-RI-C fragment of Epstein-Barr virus has been sequenced in its entirety. This same fragment has also been analyzed for RNA polymerase II promoters, which are active in a soluble in vitro assay. These data are compared to the availability of predicted open reading frames and potential nucleotide signals associated with transcription. In addition, the DNA sequence of a number of previously undetected repeated DNA sequences from this and several nearby regions of the viral genome are reported. PMID- 6296171 TI - Intermediate (10 nm) filament proteins and the Ca2+-activated proteinase specific for vimentin and desmin in the cells from fish to man: an example of evolutionary conservation. PMID- 6296172 TI - Antibodies to poliovirus detected by immunoradiometric assay with a monoclonal antibody. AB - An immunoradiometric assay (IRMA) for the assay of antibodies to poliovirus antigens is described. Dilutions of the test sera or whole (finger prick) blood samples were incubated with the Poliovirus antigen bound to a solid phase and the specific antibody was detected by the addition of a mouse anti-human IgG monoclonal antibody (McAb), which was itself revealed by iodinated sheep IgG anti mouse F(ab). We have shown that this technique is suitable for the estimation of IgG anti-poliovirus antibodies induced in children following polio vaccine. The present study shows that SPRIA provides a simple and inexpensive method for serological studies with poliovirus particularly for use in large-scale surveys. PMID- 6296173 TI - Ultrastructural localization of viral antigens using the protein A-gold technique. AB - HSV and DNV viral antigens have been localized by electron microscopy using the protein A-gold technique. The labelling of HSV antigens was detected over the (naked and enveloped) viral particles as well as on the cytoplasm and the nucleoplasm. In contrast, DNV antigens were revealed only over clusters of viral particles in the nucleus. The high sensitivity of the technique and the good ultrastructural preservation allowed a very fine identification of the labelled structures. Thus, the protein A-gold technique can be applied generally for the ultrastructural detection and identification of viral antigens and might be useful for diagnostic purposes. PMID- 6296174 TI - Sensitive interferon assay based on immunoenzymatic quantification of viral antigen synthesis. AB - A sensitive enzyme immunoassay (EIA) for determining the biological activity of human interferon was developed. Green monkey kidney (Vero) cells and human embryonic lung (HEL) cells were grown in microtitre plates, treated with leukocyte interferon (IFN alpha) and infected with vesicular stomatitis virus (VSV). Cells were fixed with paraformaldehyde and permeabilized with Triton X 100. Viral antigen synthesis was measured by labelling the cells with VSV antiserum followed sequentially by protein A horseradish peroxidase conjugate and o-phenylenediamine. Interferon activity was detected as a lowering of the absorbance value from that of the virus control wells, reflecting the inhibition of virus protein synthesis by interferon. The minimum amount of interferon producing statistically significant (P less than 0.01) decrease of absorbance in Vero cells was 1-5 international units (I.U.)/ml as in the standard plaque reduction test the detection limit was 7.5 I.U./ml or more. In HEL cells the detection limit was 1 I.U./ml measured by EIA. The EIA for interferon activity is at least as sensitive as the traditional plaque reduction test. It is reproducible, easy to automatise and requires 7-10 times less cell culture materials than the plaque reduction test. We find it preferential especially when large numbers of specimens with limited volumes are to be analysed for interferon activity. PMID- 6296175 TI - An interferon assay based on yield reduction of vesicular stomatitis virus antigen measured by enzyme immunoassay. AB - A modification of yield reduction assay for interferon was developed. The standard micromethod of inhibition of cytopathogenic effect (CPE) of vesicular stomatitis virus (VSV) was first applied and the amount of virus antigen released to the supernatant was then measured by a competitive enzyme immunoassay technique. This assay was four times more sensitive than the CPE inhibition method and was able to detect 0.50-0.25 international units/ml of human interferon-alpha and was also applicable to interferon-gamma determinations. It is a suitable method when small volumes of samples containing low levels of interferon are tested. PMID- 6296176 TI - Purification of bovine papilloma virus by gel filtration on Sephacryl S-1000 superfine. AB - Bovine papilloma virus was purified from crude extracts of bovine warts by gel filtration in Sephacryl S-1000 Superfine. Analysis by sodium dodecyl sulphate polyacrylamide gel electrophoresis and electron microscopy shows that the virus preparation is as pure as that obtained by alternative techniques. The yield of virus particles is in the range 55-80%. The main advantage of using the gel filtration technique is that it is much less time-consuming than currently used centrifugation procedures. PMID- 6296177 TI - Separation of viral DNA molecules in horizontal agarose slab gels using a new type of wick. AB - A new type of rayon fiber wick was used to establish uniform electrical paths between agarose slab gels and the electrodes of a horizontal electrophoresis apparatus. Separation of viral DNA molecules, either digested with restriction enzymes or relaxed with eukaryotic topoisomerases, was achieved easily at both low and relatively high voltage. PMID- 6296178 TI - Freeze-drying is an effective method for preserving infectious type C retroviruses. AB - Conventional freeze-drying techniques using a sucrose stabilizer and gelatin can be employed to preserve the infectivity of retroviruses. Lyophilized virus retains its infectivity even at room temperature for more than one year. A lyophilized virus preparation of Friend leukemia virus kept at 4 degrees C for more than 20 years was found to contain high titers of infectious pathogenic virus. This technique offers an easy economic means for shipping type C viruses and for preserving them for long periods of time in the laboratory without the need for cold storage. PMID- 6296180 TI - Frequency and specificity of varicella zoster virus IgM response. AB - A direct ELISA was developed for determination of IgM antibody to varicella zoster virus (VZV). With this sensitive method VZV IgM antibodies were detected in all patients with a varicella and in 84% with a herpes zoster infection. All but one of 28 renal allograft recipients had previously had varicella. A primary infection was seen in the last patient, and reactivated infections in 11 of the others. The VZV IgM response seems to be specific since patients with a herpes simplex virus (HSV) infection and a heterotypic VZV IgG titer rise did not have detectable VZV IgM. An indirect enzyme-linked immunosorbent assay (ELISA) for detection of IgG antibodies has been used for serodiagnosis of VZV infections and to determine the immune status. After injection of zoster immune globulin, it was possible to measure passively transferred antibodies. PMID- 6296179 TI - Virus-induced thymidine kinases as markers for typing herpes simplex viruses and for drug sensitivity assays. AB - A rapid, reproducible and objective new method for typing herpes simplex viruses type 1 (HSV-1) and type 2 (HSV-2) based on the effects of virus-induced thymidine kinases on various antiviral drugs has been developed. When several laboratory strains and clinical isolates were typed by this method and compared to the results obtained by the immunofluorescence antibody typing method, agreement was found for all the viruses. The new technique has the added advantage of determining the sensitivity of HSV strains to antiviral drugs. PMID- 6296181 TI - Improved yields and assay of simian varicella virus, and a comparison of certain biological properties of simian and human varicella viruses. AB - Studies were performed to define conditions under which propagation, assay and stabilization of the Delta herpesvirus (DHV) strain of simian varicella virus might be improved, and to compare biological properties of DHV with those of human varicella zoster virus (VZV). A mycoplasma contaminant was successfully eliminated from the DHV seed virus by treatment with a specific anti-serum. DHV was found to replicate more efficiently in the BS-C-1 line of African green monkey kidney cells than in Vero cells, and seed virus preparations in the form of virus-infected cells were produced which had infectivity titers greater than or equal to 1 X 10(6) p.f.u./ml. Greater yields of virus were produced in cultures infected as dispersed cells than as preformed monolayers. Infectious DHV could be released from host cells by sonic treatment of heavily infected cultures at 48 h post infection. Certain agents reported to enhance replication of herpes viruses (caffeine, carbaryl, the tumor promoter 12-0-tetra-decanoyl-phorbol-13 acetate, and DEAE-dextran) had no enhancing effect on replication of DHV. However, DEAE-dextran in the maintenance medium enhanced spontaneous release of DHV into culture fluids. Plaquing efficiency and plaque size of DHV were greater in BS-C-1 than in Vero cells, and plaque assays and plaque reduction neutralization tests were developed in this cell system using a solid overlay medium with neutral red vital stain. Neutralization of DHV was markedly enhanced by fresh guinea pig complement. The newly developed neutralization test demonstrated more vigorous antibody responses to DHV in active and latent VZV infections than were demonstrated with previous procedures. In addition to their preferential growth in monkey and human cells respectively, DHV and VZV were found to differ markedly in their rates of attachment to host cells, with DHV requiring over 6 h of adsorption, while VZV adsorption was essentially complete at 1 h. Also, cell-free DHV was much more resistant than cell-free VZV to repeated cycles of freezing and thawing. PMID- 6296182 TI - Clinical and biochemical variability of congenital adrenal hyperplasia due to 11 beta-hydroxylase deficiency. A study of 25 patients. AB - Twenty five patients (10 males and 15 females) aged 0-23 yr with congenital adrenal hyperplasis due to 11 beta-hydroxylase deficiency were studied. They were divided into 13 classic (group A), and 12 mild (group B) patients. The patients of group A were diagnosed at a younger age and had more severe clinical symptoms (ambiguous genitalia in girls, pseudoprecocious puberty in boys). Two had neonatal salt wasting before treatment, and one gynecomastia. Seven had moderate to severe hypertension. Their mean 3 alpha,17,21-trihydroxy-5 beta-pregnan-20-one (THS) and 3 alpha, 21-dihydroxy-5 beta-pregnane-11,20-dione (THDOC) excretion was 14.2 +/- 4.1 and 7.2 +/- 4.2 mg/m2 . day, respectively. The patients of group B had mostly late onset of symptoms (hirsutism, amenorrhea in girls, pseudoprecocious puberty in boys, tall stature, and advanced bone age in both sexes). One boy had bilateral cryptorchidism. Four had moderate hypertension. In seven patients, THS (5.3 +/- 2.3 mg/m2 . day) and THDOC (3.9 +/- 0.5 mg/m2 . day) responded to ACTH. In five, only THS (4.3 +/- 1.1 mg/m2 . day) responded, but THDOC remained undetectable. It is concluded that the clinical and biochemical expression of 11 beta-hydroxylase deficiency is variable, that hypertension in not directly related to deoxycorticosterone, and that, regardless of the intensity of the defect, there are patients in whom the 11 beta-hydroxylation of 17 alpha-hydroxylated steroids only is impaired, and others in whom both the conversion of 17,20-dihydroxy-4-pregnene-3,20-dione and deoxycorticosterone are reduced. PMID- 6296183 TI - Reciprocal changes in active and inactive renin after converting enzyme inhibition in normal man. AB - Since one mechanism by which converting enzyme inhibition (CEI) increases renin is removal of angiotensin II negative feedback on the juxtaglomerular cell, we studied the time course of changes in active and inactive renin after CEI. After equilibration on a 25 meq/day sodium diet, captopril was given as a single 50-mg oral dose (acute phase), and then was administered as 50 mg every 6 h for 3 days to seven normal volunteers (chronic phase). In the acute phase, supine blood pressure fell 12 +/- 2 mm Hg (P less than 0.02). Active renin acutely increased 12.5 +/- 0.9 times the baseline value, peaking at 3-4 h. Inactive renin, measured by acid activation of trypsin activation, decreased in all subjects to 10% or less of control from 2 to as long as 6 h post-CEI and then returned to baseline levels by 8 h (P less than 0.01). With chronic CEI, active renin was elevated to 10.8 +/- 2.4 times the baseline level, and after 48 h inactive renin levels rose to 4.0 +/- 0.6 times the baseline (P less than 0.02). To determine whether the acute changes in inactive and active renin occurred because of captopril's effect on renin in the circulation or kidney, a single dose of captopril was administered to three subjects with mild to moderate renal insufficiency and hyporeninemic hypoaldosteronism. In contrast to normal subjects, these patients had no change in active and inactive renin levels when given captopril, suggesting that changes observed in the normals were renal mediated rather than a plasma phenomenon. We conclude that CEI 1) acutely increases active renin while reciprocally reducing the inactive form, and 2) chronically increases both active and inactive renin. These studies support the hypothesis that inactive renin may be a precursor of circulating active renin. PMID- 6296184 TI - Adenosine amplifies follicle-stimulating hormone action in granulosa cells and luteinizing hormone action in luteal cells of rat and human ovaries. AB - It is known that adenosine amplifies LH-stimulated cAMP accumulation and progesterone production in rat luteal cells. The aim of this study was to investigate the effect of adenosine on cAMP and steroid production in short term cultures of rat and human luteal cells in the presence of LH and in rat and human granulosa cells in the presence of FSH. In rat luteal cells, the maximal cAMP response to LH and adenosine amplification of this response occurred in the midluteal phase and was decreased in both early and late luteal cells. In humans, adenosine (50 microM) increased cAMP and and progesterone accumulation by 100% and 40%, respectively, in periovulatory granulosa cells. Adenosine also amplified cAMP accumulation in response to increasing hCG concentrations by 2- to 3-fold in human luteal cells. The ability of the human luteal cell to respond to hCG with cAMP accumulation and the ability of adenosine to amplify this cAMP response appeared to be inversely related to human luteal cell age. In isolated preparations of rat granulosa cells, adenosine amplified cAMP accumulation in response to FSH, and cAMP accumulation was inversely proportional to the duration of follicular development. In human periovulatory granulosa cells, adenosine (100 microM0 increased cAMP accumulation by 2-fold and amplified FSH-stimulated cAMP accumulation by 25%. These studies suggest that in both the rat and human, adenosine may physiologically affect gonadotropin function in both follicular and luteal ovarian tissue. PMID- 6296185 TI - Metabolic and blood pressure responses to hydrocortisone in the syndrome of apparent mineralocorticoid excess. AB - A syndrome of low renin hypertension in childhood with apparent mineralocorticoid excess associated with a defect in the peripheral metabolism of cortisol has been described previously in 2 patients. In these patients, decreased secretion rates of glucocorticoids, mineralocorticoids, and sex steroids have been demonstrated. In a 10(10/12)-yr-old girl with this disorder, continuous iv administration of hydrocortisone in doses of 5, 10, 15, and 20 mg/day resulted in an increase in blood pressure and a decrease in serum potassium concentration. The addition of spironolactone during the continued administration of 20 mg/day hydrocortisone did not result in a decrease in blood pressure. Withdrawal of hydrocortisone and continued administration of spironolactone alone resulted in a decrease in blood pressure, a rise in serum potassium concentration, and a fall in serum sodium concentrations. These studies suggest that an abnormality in cortisol action or metabolism causing cortisol to behave as a potent mineralocorticoid may account for this syndrome of apparent mineralocorticoid excess. PMID- 6296186 TI - Cyproheptadine and mineralocorticoid effector mechanisms. AB - Cyproheptadine has recently been reported to blunt the furosemide-induced rise in PRA in normal subjects and to acutely lower plasma aldosterone levels in patients with hyperaldosteronism due to bilateral adrenal hyperplasia; both actions have tentatively been ascribed to the antiserotoninergic action of the drug. We here describe receptor studies showing cyproheptadine to occupy mineralocorticoid, but not glucocorticoid, receptors in rat and mouse kidney. On bioassay, cyproheptadine is a partial mineralocorticoid agonist/predominant antagonist. PMID- 6296187 TI - Studies on the regulation of phosphodiesterase in human adipose tissue in vitro. AB - The influence of several agents known to affect the rate of lipolysis or the phosphodiesterase (PDE) activity in rat adipose tissue was studied in human adipose tissue in vitro. Only insulin, catecholamines, prostaglandin E1, and cAMP increased PDE activity in man (by 20-40% over the control values). In dose response studies, it was found that the ED50 values for insulin and isopropyl noradrenaline were of the same magnitude as the previously observed ED50 values for lipolysis in human adipose tissue. The time course for insulin and isopropyl noradrenaline showed maximum stimulation after 10 min, followed by a decline, with the curve approaching zero at 60 min. In subcellular fractions of adipose tissue, the highest specific PDE activity was found in the particulate fraction, in which the effects of both insulin and isopropyl noradrenaline were most marked. It is concluded that the stimulatory effect of catecholamines on PDE may be one reason for the failure of these agents to produce a sustained increase in the cAMP level and that the effect of insulin on PDE may be one mechanism by which insulin reduces the rate of lipolysis. PMID- 6296188 TI - Chronic idiopathic polyhydramnios: evidence for a defect in the chorion laeve receptor for lactogenic hormones. AB - Binding of human GH (hGH) to the lactogenic receptor of human chorion laeve has been compared in membrane preparations from normal pregnancies and those complicated by chronic idiopathic polyhydramnios. Specific binding of [125I]hGH was significantly lower (mean +/- SE, 1.64 +/- 0.28%; n = 5) in chronic idiopathic polyhydramnios than in normal pregnancies (2.93 +/- 0.4%; n = 16). Scatchard analysis data were consistent with a reduced lactogenic hormone receptor concentration being the explanation for this reduced hGH binding. By contrast, the specific binding of [125I]insulin to its receptor was unchanged when chorion laeve from hydramniotic pregnancies (7.29 +/- 1.98%; n = 4) was compared with that from normal pregnancies (8.63 +/- 1.35%; n = 7). We conclude that a lactogenic hormone receptor defect exists in the chorion laeve of pregnancies complicated by chronic idiopathic polyhydramnios. Such impaired binding for PRL may explain the development of excessive amniotic fluid volumes, which is characteristic of this complication of pregnancy. PMID- 6296189 TI - Normalization of androgen and sex hormone-binding globulin levels after treatment of hyperprolactinemia. AB - Twenty-eight women with amenorrhea, galactorrhea and hyperprolactinemia without hirsutism were studied before and after bromocriptine therapy for 2 months. Compared to 15 euprolactinemic controls, hyperprolactinemic women had elevated levels of dehydroepiandrosterone sulfate and androstenedione and lower levels of total testosterone (T), and androst-5-ene-3 beta, 17 beta-diol (Adiol), and 17 beta-estradiol (P less than 0.05). Unbound T and unbound Adiol were significantly elevated, while sex hormone-binding globulin binding capacity was decreased (P less than 0.05) and corticosteroid-binding globulin binding capacity was normal. After treatment with bromocriptine, dehydroepiandrosterone sulfate and androstenedione decreased to control levels, as did unbound Adiol, while 17 beta estradiol and sex hormone-binding globulin binding capacity levels increased significantly (P less than 0.05). Five hyperprolactinemic women underwent ACTH stimulation tests before and after treatment, and the results were compared to those of seven controls. Steroid ratios in response to ACTH suggested normal 3 beta ol-dehydrogenase-isomerase, 17-20-desmolase and 17 beta-hydroxysteroid dehydrogenase enzymatic activities in hyperprolactinemia. Basal steroid ratios of T to 5 alpha-androstane-17 beta-01-3-one) (DHT) and of unbound T to unbound dihydrotestosterone were elevated (P less than 0.05), suggesting reduced 5 alpha reductase activity in hyperprolactinemia which is normalized after treatment. Hirsutism was not present in these patients with hyperprolactinemia despite elevated levels of unbound T and Adiol, and may be explained by reduced 5 alpha reductase activity. Our data suggest that the increased levels of androgens in these patients result from the hyperprolactinemia. PMID- 6296190 TI - The effect of chronic metabolic acidosis on the end organ responsiveness to parathyroid hormone in man. AB - To examine the effect of prolonged metabolic acidosis on the responsiveness of end organs to PTH, we measured the changes in serum ionized calcium and the urinary excretion of calcium and phosphorus in response to a 10-h infusion of parathyroid extract (PTE) before and after 9 days of metabolic acidosis. During the first hours of PTE infusion, the increases in ionized calcium were slower during acidosis, but the maximal increments in ionized calcium (0.5 +/- 0.1 and 0.4 +/- 0.1 mg/dl, respectively) were not different. Moreover, the PTE-induced increments in the fractional excretion of phosphate during acidosis and the control study were not different. Fractional excretion of calcium before the infusion of PTE was higher in acidosis than in control (3.7 +/- 0.3% compared to 0.8 +/- 0.3%; P less than 0.01). However, at 10 h of the infusion, when the maximal effect of PTE was observed, the fractional excretions of calcium in control and acidosis were not significantly different. These data suggest that prolonged metabolic acidosis has little or no effect on bone or renal responsiveness to PTH. PMID- 6296191 TI - Failure of angiotensin II to stimulate increases in concentrations of adrenal androgens, 17-hydroxyprogesterone, or adrenocorticotropin in congenital 21 hydroxylase deficiency. AB - To determine if angiotensin II stimulates an increase in the plasma concentration of androstenedione, dehydroepiandrosterone, 17-hydroxyprogesterone, or ACTH in a patient with congenital 21-hydroxylase deficiency, we measured these plasma concentrations before and after the plasma angiotensin II concentration was increased by upright posture and angiotensin II infusion in a surgically castrate XX adult patient with this disorder. The patient was studied before treatment, after treatment with 1 mg dexamethasone daily for 3 weeks, and after treatment with both dexamethasone and 0.2 mg fludrocortisone daily for 3 weeks. The plasma concentrations of androstenedione, dehydroepiandrosterone, and 17 hydroxyprogesterone did not change consistently during increases in the angiotensin II concentration. The ACTH concentration did not increase in response to raised angiotensin II concentrations before or after steroid treatment. During the infusion of angiotensin II, blood pressure increased and renin activity decreased appropriate in degree to the preinfusion concentration of angiotensin II. The results from the study of this patient do not support the hypotheses that in congenital 21-hydroxylase deficiency, angiotensin II directly stimulates adrenal androgen secretion or that angiotensin II stimulates ACTH secretion. PMID- 6296192 TI - Enzyme immunofiltration technique for rapid diagnosis of herpes simplex virus eye infections in a rabbit model. AB - A rapid enzyme immunofiltration assay for herpes simplex virus (HSV) has been developed which is sensitive enough to detect viral antigens in eye swabs from rabbits with primary herpes keratitis. This assay employs a specially designed filter manifold to immobilize whole cells and cell debris dissociated from the swabs. Viral antigens trapped on the filters are then detected in an indirect immunoassay utilizing staphylococcal protein A conjugated with horseradish peroxidase. The assay required only 2.5 h to perform and could be read visually. Reconstruction experiments indicated that antigen from as few as 49 HSV-infected cells could be detected. Calcium alginate swabs were shown to recover more viral antigen than dacron swabs. The enzyme immunofiltration assay detected HSV antigens on 95% of the eye swabs from which infectious virus was recovered. In addition, HSV antigen was also detected in several swabs from infected eyes which did not yield infectious virus, presumably because the virus was neutralized by native antibody present in the lacrimal fluid. This enzyme immunofiltration assay technique lends itself to the elution of native antibody bound to the viral antigens, and this may be especially applicable in the diagnosis of recurrent HSV keratitis, where antiviral antibody in the lacrimal fluid may interfere with virus isolation and fluorescent-antibody or other virus detection assays. PMID- 6296193 TI - Rotavirus and hemolytic enteropathogenic Escherichia coli in weanling diarrhea of pigs. AB - Since the turn of the century, Escherichia coli has been implicated in the etiology of weanling diarrhea (colibacillosis). However, rotavirus--a virus that destroys enterocytes--has been shown recently to be causally associated with weanling diarrhea of pigs. The role of both rotavirus and hemolytic enteropathogenic E. coli in weanling diarrhea was assessed in this study. Pigs from a closed herd were farrowed and weaned by two markedly different systems: an "intensive care sanitary" system and a "conventional unsanitary" system. Pigs weaned at 3 weeks of age in the sanitary system usually experienced a rotaviral diarrhea about 16 days postweaning. No hemolytic E. coli were detected in feces from these pigs. Peers weaned at the same time by the unsanitary system commenced diarrhea 3 days postweaning. Rotavirus and nonhemolytic E. coli were detected in the feces at the onset of diarrhea and for a few days thereafter. Then, the aerobic fecal flora shifted to nearly pure hemolytic enteropathogenic E. coli. About 10 days later, the diarrhea waned, and the fecal flora shifted back to nonhemolytic E. coli. This hemolytic E. coli shedding pattern could not be duplicated in artificially inoculated sanitary pigs unless they were inoculated with the hemolytic E. coli during a rotaviral-associated diarrhea. Otherwise, the shedding of hemolytic E. coli was fleeting, and the diarrhea, if present, was mild. Pigs developed humoral antibodies to the rotavirus but not to the hemolytic E. coli. We conclude that rotavirus damages the epithelium of the small intestines, which changes the luminal environment to one that favors colonization by enteropathogenic E. coli. PMID- 6296194 TI - Detection of a rotavirus-like agent associated with diarrhea in an infant. AB - A rotavirus-like agent was detected in the feces of a child with diarrhea. Although morphologically indistinguishable from rotavirus, the agent was serologically distinct, and electrophoresis of its nucleic acid also showed that it was dissimilar. PMID- 6296195 TI - Isolation of human rotavirus subgroups 1 and 2 in cell culture. AB - One strain of human rotavirus subgroup 1 (KUN) and one strain of subgroup 2 (MO) were isolated with the MA104 cell line, a fetal rhesus monkey kidney cell line. Their subgroup specificities and RNA patterns were identical to those of rotaviruses present in stools before cultivation. Distinct cytopathic effects consisting of obscure cell boundaries, cell fusion, cell rounding, cell detachment, and lytic foci were recognized at passage 3 of MO and passage 6 of KUN. No differences in cytopathic changes were found between the two isolates. PMID- 6296196 TI - Clinical laboratory evaluation of a reverse CAMP test for presumptive identification of Clostridium perfringens. AB - Ninety-six percent of Clostridium perfringens isolates from clinical specimens were reverse CAMP test positive, whereas several other Clostridium species tested were reverse CAMP test negative. C. perfringens was detected by direct inoculation of clinical specimens to reverse CAMP plates, and the reverse CAMP procedure provided reliable presumptive identification of this organism. PMID- 6296198 TI - Survival of hepatitis A virus in feces after drying and storage for 1 month. AB - Hepatitis A virus in feces remained viable after being dried and then stored at 25 degrees C and 42% relative humidity for 30 days, as evidenced by infection of two marmosets inoculated with 1 ml of the treated fecal material. The virus was excreted in stool specimens from these animals, and seroconversion to antibody to hepatitis A virus occurred 28 and 35 days post-inoculation. PMID- 6296197 TI - Immunity to rotavirus in conventional neonatal calves. AB - The local and systemic humoral immune responses to rotavirus were studied in six conventional neonatal calves. Attenuated bovine rotavirus was administered either orally or directly into an isolated intestinal loop. The parameters monitored were neutralizing rotavirus antibody in serum, immunofluorescent and neutralizing rotavirus antibody in intestinal loop washings, and rotavirus antibody-producing cells in intestinal mucosa. An antibody response was observed in the serum and intestinal secretions from one calf only. Viral replication was not detected in the isolated intestinal loop. Rotavirus antibody-producing cells were found in the intestinal mucosa of five calves. Double staining revealed that most of these cells produced antibody of the immunoglobulin A class. The conclusions were: (i) a previously described system to detect rotavirus antibody-producing cells can be used to study immune responses in neonatal calves, (ii) the class or subclass of antibody in rotavirus antibody-producing cells can be determined by double immunofluorescent staining, (iii) neonatal calves respond to rotavirus inoculation with a local immunoglobulin A response, and (iv) most of the rotavirus antibody-producing cells are located in the mucosa of the proximal small intestine. PMID- 6296199 TI - Different virus antibodies in serum and cerebrospinal fluid of patients suffering from subacute sclerosing panencephalitis. AB - Complement fixing (CF) antibody titers to measles, parainfluenza (PI) types 1 and 3, mumps, herpes type 1, and cytomegalovirus (CMV) in serum and cerebrospinal fluid (CSF) of 33 patients with subacute sclerosing panencephalitis (SSPE) were evaluated. Results were analyzed in comparison to 11 patients with neurological diseases other than SSPE and 7 normal subjects. All SSPE patients had elevated serum and CSF measles antibody titers. The number of SSPE patients manifesting elevated titers other than measles did not reach statistical significance when compared to controls, except for PI type 1. This suggests a possible dual infection with measles and PI in SSPE. The anticomplementary effect detected in the serum and CSF of some patients indirectly suggests the presence of immune complexes. PMID- 6296200 TI - Resistance to 1,25-dihydroxyvitamin D. Association with heterogeneous defects in cultured skin fibroblasts. AB - We evaluated the interaction of [3H]1,25(OH)2D3 with skin fibroblasts cultured from normal subjects or from affected members of six kindreds with rickets and resistance to 1-alpha, 25(OH)2D [1,25(OH)2D]. We analyzed two aspects of the radioligand interaction; nuclear uptake with dispersed, intact cells at 37 degrees C and binding at 0 degrees C with soluble extract ("cytosol") prepared from cells disrupted in buffer containing 300 mM KCl and 10 mM sodium molybdate. With normal fibroblasts the affinity and capacity of nuclear uptake of [3H]1,25(OH)2D3 were 0.5 nM and 10,300 sites per cell, respectively; for binding with cytosol these were 0.13 nM and 8,900 sites per cell, respectively. The following four patterns of interaction with [3H]1,25(OH)2D3 were observed with cells cultured from affected patients: (a) two kindreds; cytosol binding and whole-cell nuclear uptake both unmeasurable; (b) one kindred, decreased capacity and normal affinity both for binding in cytosol and for nuclear uptake in whole cells; (c) two kindreds, normal or nearly normal capacity and affinity of binding in cytosol but unmeasurable whole-cell nuclear uptake; and (d) one kindred, normal capacity and affinity of both cytosol binding and whole-cell nuclear uptake. In all cases where the radioligand bound with high affinity in nucleus or cytosol, the nucleus- or cytosol-associated radioligand exhibited normal sedimentation velocity on sucrose density gradients. When two kindreds exhibited similar patterns (i.e. pattern a or c) with the analyses of cultured fibroblasts, clinical features in affected members suggested that the underlying genetic defects were not identical. IN CONCLUSION: (a) Fibroblasts cultured from human skin manifest nuclear uptake and cytosol binding of [3H]1,25(OH)2D3 that is an expression of the genes determining these processes in target tissues. (b) Based upon data from clinical evaluations and from analyses of cultured fibroblasts, severe resistance to 1,25(OH)2D resulted from five or six distinct genetic mutations in six kindreds. PMID- 6296201 TI - Leupeptin inhibits adrenocorticotropic hormone-induced protein breakdown in the conscious dog. AB - To elucidate the role of proteinase inhibitors in the regulation of protein breakdown in vivo, we measured the effect of leupeptin on the rate of appearance of leucine in the plasma compartment in overnight-fasted conscious dogs. Two groups of dogs were studied. The control group (I) received saline infusion, and the experimental group (II) was rendered hypercatabolic with daily administration of adrenocorticotropic hormone (ACTH) (500 U/d) for 4 d.ACTH treatment increased plasma cortisol from 2+/-0.4 to 17+/-2 mug/dl (P < 0.005). It raised plasma leucine levels (mumol/liter) from 123+/-6 in I to 206+/-5 in II (P < 0.01) and its rate of appearance into the plasma compartment (micromoles per kilogram per minute) from 3.1+/-0.1 in I to 4.6+/-0.3 in II (P < 0.01). Whole blood alanine concentration (micromoles per liter) increased by 50% (from 387+/-31 to 577+/-53, P < 0.01) and whole blood glutamine concentration (micromoles per liter) increased from 653+/-51 to 917+/-93 (P < 0.01). Leupeptin infusion in the ACTH treated group significantly decreased both the concentration of plasma leucine and its rate of appearance. Blood glutamine declined by 30% (P < 0.05) after leupeptin, but no effect on blood alanine was observed. Leupeptin had no effect on the saline control group. These data indicate that leupeptin decreases the accelerated rate of protein breakdown induced by cortisol excess. The fact that it did not affect protein degradation in controls may indicate that control of protein breakdown in the postabsorptive state may differ from that during accelerated turnover. Thus, the antibiotic proteinase enzyme inhibitors may be potentially useful in treating conditions of inappropriate protein breakdown. PMID- 6296202 TI - Circulating bovine lymphocytes contain receptors for parathyroid hormone. AB - No cell type practicably obtainable in vivo, such as blood cells, is known to contain parathyroid hormone (PTH) receptors; this deficiency has hampered investigation of receptor regulation. Second, PTH in vivo is among the potent stimulators of osteoclastic activity, although no direct hormonal effects on these cells have been identified. Several lines of evidence suggest that cells of the immune system may mediate PTH effects on osteoclasts. We, therefore, studied bovine blood cells for the presence of PTH receptors and PTH-stimulated adenylate cyclase. Using an analogue of bovine PTH, (125)I-labeled [Nle(8),Nle(18),Tyr(34)]bPTH-(1--34)amide, we found PTH-specific binding to intact, nonadherent mononuclear cells (lymphocytes) and PTH-stimulated adenylate cyclase in plasma membranes prepared from these cells, and not with cells or membranes from other blood cells. Lymphocytes may serve to study the effects of physiologic and pathologic perturbations on PTH-receptor function in vivo. Exploration of PTH-related lymphocyte responses may help define the relation between cells of the immune system and osteoclastic bone resorption. PMID- 6296203 TI - Urine cyclic nucleotides in cancer and other conditions. PMID- 6296205 TI - Variation in the thermal stability of isolates of foot-and-mouth disease type SAT 2 and its significance in the selection of vaccine strains. PMID- 6296207 TI - Electron microscopy of the intestine of gnotobiotic piglets infected with porcine rotavirus. PMID- 6296204 TI - Vaccination trials against pigeon herpesvirus infection (Pigeon herpesvirus 1). PMID- 6296206 TI - Establishment of a chicken lymphoblastoid cell line infected with reticuloendotheliosis virus. PMID- 6296208 TI - Presumptive adenoviral bronchiolitis in a red deer (Cervus elaphus). PMID- 6296209 TI - Bovine leukosis--its importance to the dairy industry in the United States. AB - Bovine leukosis describes lymphatic cancers of cattle. The most common form of this disease occurs in adult animals and is caused by bovine leukemia virus. Infection is widespread in the United States, especially in dairy cattle, but the virus produces tumors in only a small percentage of infected animals. Nevertheless, bovine leukemia virus has been receiving attention from the dairy industry because of its importance in health certification of cattle or semen intended for export. Another source of concern is whether bovine leukemia virus poses any risk to human health. These problems are discussed in the light of recent technological advances in tumor virus research and specifically regarding our current understanding of the biology of bovine leukemia virus. PMID- 6296210 TI - Experimental ridge augmentation with porous hydroxyapatite implants. AB - A new hydroxyapatite ceramic implant material for restoration of atrophic edentulous ridges was tested in a canine animal model. The structure of this material is unique, in that it is produced from the skeleton of a coral, and in that it is penetrated by a uniform network of interconnected pores. Results indicate that bone penetrates its structure to a greater degree than had been observed with similar implants constructed of different materials. Implant insertion through a simple surgical technique involving minimal morbidity is a viable method of placement. PMID- 6296211 TI - Microbeam x-ray diffraction analysis of dental calculus. AB - The crystalline components of human dental calculus were investigated using microbeam x-ray diffraction analysis. Hydroxyapatite and octacalcium phosphate were most frequently found in that portion having porous and zonal structure. In the portion of the homogeneous illustration showing high calcification, whitlockite is a main component. Brushite was unexpectedly rare, and no calcite was detected in any portion of human dental calculus. The mechanism of the formation of dental calculus being considered in this paper is that octacalcium phosphate or brushite is formed during the initial stage of calcification of dental plaque, and is gradually hydrolyzed and transformed into hydroxyapatite and/or whitlockite. PMID- 6296212 TI - Correction of alveolar ridge deficiencies with nonresorbable hydroxylapatite. PMID- 6296213 TI - Intervention with epinephrine in hypotension associated with mastocytosis. AB - The occurrence of the episodes of vasodilatory hypotension can be a life threatening manifestation of systemic mastocytosis. This article describes the reversal by epinephrine of episodes of severe hypotension in two hospitalized patients with mastocytosis. Recognition of the efficacy of epinephrine in hypotension associated with mastocytosis can be important when other methods fail to restore hemodynamic stability. PMID- 6296214 TI - Immunologic cross-reactivity between different albumin-bound isocyanates. AB - Sera of six workers with conclusive evidence for IgE-mediated sensitization to isocyanates were used for evaluation of immunologic cross-reactivities among eight different isocyanate-protein conjugates. In all cases RAST and/or skin-test investigations revealed the presence of IgE antibodies reacting specifically with HSA conjugated with those isocyanates to which workers were exposed as well as with other isocyanates with which they had not been in contact. By the RAST inhibition technique, moderate to strong mutual cross-reactivities between all tested isocyanate-HSA conjugates--even between aromatic and aliphatic ones--could be demonstrated in tests with five sera. The magnitudes of cross-reactivities differed, however, from one patient to another. One serum contained IgE antibodies that were almost completely specific to TDI-HSA; with this serum only weak cross-reactivities with other isocyanate conjugates could be demonstrated. These results indicate the predominance of closely related antigenic determinants in HSA conjugated with different isocyanates. The common antibody-binding regions are obviously recognized to different extents by antibodies of clinically sensitized workers, indicating individual differences in specificities and avidities of antibody populations. Nearly complete lack of IgE binding of ovalbumin-bound TDI in RAST and RAST inhibition indicates carrier-specific antigenicity of isocyanate-protein conjugates. In addition, since unmodified HSA did not bind IgE, antigenic determinants of the conjugates studied should be predominantly formed by the isocyanate-protein bond regions and concurrently by neighboring amino acid residues of the HSA molecule. PMID- 6296215 TI - Difference between lung and spleen susceptibility of beta-adrenergic receptors to desensitization by terbutaline. AB - There are many reports about a marked down-regulation of beta-adrenergic receptors by beta agonists in human leukocytes. To determine whether beta receptors in lung tissue are down regulated by the long-term administration of beta agonists, as are those in the spleen (which consists largely of lymphocytes), we injected terbutaline (0.1 mg/kg, t.i.d.) intramuscularly into rats for either 3 or 6 days. We observed a significant decrease in beta-receptor density in spleen tissue but not in lung parenchyma, associated with terbutaline injection. The affinity for an antagonist did not change significantly in any group in either tissue. We did not observe any change in alpha 1-adrenergic receptors in the lung after this treatment. In in vitro studies, we also observed reduced beta-receptor density in spleen cells but not in lung parenchyma after incubation of these tissues with terbutaline. However, there was agonist-specific alteration in lung beta receptors. It was found that the isoproterenol competition curve for 3H-dihydroalprenolol binding shifted to the right and steepened, suggesting reduced affinity of the receptors for isoproterenol. We used whole lung and did not examine bronchial smooth muscle per se, nor were functional studies performed. Our results show a difference between lung parenchyma and spleen tissue in the susceptibility of beta receptors to desensitization by an adrenergic agonist and suggest that there may be such a difference in sensitivity between beta receptors in human lung and leukocytes. PMID- 6296216 TI - [Lafora bodies in the retina of various animals]. AB - Lafora's bodies, formerly described in the retina in humans affected with familial myoclonic epilepsy--or Lafora's disease--and in dogs, were found not only in retinas of clinically healthy monkeys (macacus rhesus), but also in those of the healthy cat and dog. Lafora's bodies are not there fore characteristic of the specific disease. PMID- 6296217 TI - [Value of the immunoenzyme method (ELISA) in determining serum and aqueous humor antiherpes immunoglobulin levels (apropos of 155 cases)]. AB - Immuno-enzymatic assay of aqueous humor and serum antiherpetic antibodies was performed in 155 patients with all types of uveitis and with keratitis. Globulin levels were also determined in the two fluids, employing an original nephelometry laser technique. This immuno-enzymatic technique, which possesses greater reliability, reproducibility, and sensitivity than passive hemagglutination, appears suitable for microassay in aqueous humor, and demonstrates the production of anti-herpes antibodies in this fluid to the exclusion of other specificity. Applying limits of at least 1/40e for H.A. antibody levels, and at least 10 for the immunity load coefficient, these antibodies could be demonstrated in half of the cases of clinically confirmed herpes, in one-third of clinically suspected cases, and in two intermediary cases of uveitis where no predictive signs of herpes were present. In contrast, specific antibodies were never detected in 70 cases of anterior or total uveitis. Measuring anti-herpes antibodies in aqueous humor and serum by the ELISA method, in association with immunity load coefficients determination in the two fluides, appears to be a very useful method for the future etiological diagnosis of anterior and intermediary uveitis when the etiology is uncertain but clinical signs suggest a possible herpetic origin. PMID- 6296218 TI - In vitro steroidogenic properties of FK 33 824, a stable analog of methionine enkephalin. Opiate-dopamine interaction in the control of aldosterone production. AB - The steroidogenic properties of a stable analog of the endogenous opioid methionine-enkephalin, FK 33 824, were studied with calf adrenal glomerulosa cells and its effects were compared to those of angiotensin II (A II) and metoclopramide. Metoclopramide, A II, and FK 33 824 induced dose-related increases in aldosterone production. The order of potency in stimulating aldosterone was A II, FK 33 824, metoclopramide. Metoclopramide and FK 33 824 did not increase cortisol production. The response to A II but not to FK 33 824 was inhibited by equimolar concentrations of (Sar1 Ala8) antagonist analog of AII (saralasin acetate). By contrast in the presence of equimolar concentrations of naloxone, an opioid receptor antagonist, FK 33 824-induced aldosterone production was markedly inhibited while the response to A II was unchanged. Increases in cAMP accompanied the steroidogenic response to ACTH but not to A II or FK 33 824. Dopamine at physiological concentrations (10(-10) M) inhibited FK 33 824-induced aldosterone production. These results suggest that FK 33 824 is an aldosterone secretagogue and that it initiates steroidogenesis by mechanisms similar to those of A II. However the inability to block its effect with a specific antagonist of A II provides evidence for its action on a separate site. PMID- 6296219 TI - Monoclonal antibodies to the thyrotropin receptor: the identification of blocking and stimulating antibodies. AB - Monoclonal antibodies to the thyrotropin (TSH) receptor have been obtained from fusions of mouse myeloma cells with spleen cells immunized with solubilized thyroid membrane preparations. Two monoclonal antibodies which inhibit 125I-TSH binding and are reactive with the glycoprotein component of the bovine TSH receptor (11E8 and 13D11), are shown to inhibit basal and TSH stimulated adenylate cyclase activity in bovine thyroid membranes and human thyroid cells. Both antibodies also inhibit 125I-TSH binding in vitro, whether binding is measured at pH 6.0 in low salts and at 0-4 C or at pH 7.4 in 50 mM NaCl and at 37 C. The glycoprotein component is thus a portion of the physiologic TSH receptor in vivo and 125I-TSH binding studies apparently measure the high affinity glycoprotein component under nonphysiologic conditions and conditions more representative of the physiologic milieu. A third monoclonal antibody whose interaction with thyroid membranes is prevented by TSH is shown to stimulate adenylate cyclase activity in bovine thyroid membranes and human thyroid cells. This stimulating antibody only weakly inhibits 125I-TSH binding to thyroid membranes or to the glycoprotein component of the TSH receptor. The 22A6 antibody does, however, immunoprecipitate mixed brain gangliosides, in distinct contrast to the monoclonal antibodies to the glycoprotein receptor component, i.e., 11E8 and 13D11. The results support the speculation that autoimmune antibodies which inhibit TSH binding to thyroid membranes are not necessarily identical to antibodies which stimulate function; that antibodies directed at the high affinity initial site of TSH interaction with a cell can behave as blocking rather than stimulating antibodies and that a possible relationship exists between stimulating antibodies and the low affinity TSH binding sites (gangliosides) on thyroid membranes. PMID- 6296220 TI - Portal, hepatic and peripheral insulin immunoreactive substances before and after removal of an insulinoma. AB - Plasma insulin immunoreactivity (IRI) results from high molecular weight substances with insulin immunoreactivity (HWIRI), proinsulin (PI) and insulin (I). Their respective concentrations and percentages of IRI were determined preoperatively in the hepatic and peripheral circulations and after surgery in the latter two and in the portal circulation in a patient with a pancreatic adenoma. Removal of the insulinoma resulted in a highly significant reduction of PI concentration in the hepatic (from 16.2 +/- 1.9 microU/ml to 7.3 +/- 0.8 microU/ml) and in the peripheral (from 24.0 +/- 2.0 microU/ml to 9.2 +/- 1.2 micro U/ml) venous systems. Similarly, I concentration decreased in the hepatic (from 36.9 +/- 3.4 microU/ml to 28.0 +/- 1.0 microU/ml) and peripheral (from 33.8 +/- 3.4 microU/ml to 19.8 +/- 2.0 microU/ml) venous systems. HWIRI concentration decreased in the hepatic venous system (from 6.9 +/- 0.8 microU/ml, to 4.8 +/- 0.6 microU/ml), but the decrease in the peripheral venous system (from 9.7 +/- 1.4 microU/ml to 8.2 +/- 1.2 microU/ml) was not significant. It was found that the concentration of I virtually was the same in hepatic and peripheral blood prior to operation in contrast to a decrease seen after the operation. This indicates that the hyperinsulinemia caused primarily by the autonomous hypersecretion of the insulinoma was amplified by a decreased degradation of insulin by the liver and kidney. Our results also indicate that HWIRI was found with the highest concentration in the peripheral blood independently of the adenoma. PMID- 6296221 TI - Unconventional application of standard light and electron immunocytochemical analysis to aldehyde-fixed, araldite-embedded tissues. AB - A simple procedure for the immunocytochemical analysis of glutaraldehyde/formaldehyde-fixed, Araldite- or Epon-embedded tissues by either light or electron microscopy is presented. Retention of immunoreactive antigen in deplasticized sections was achieved by use of a low concentration of glutaraldehyde in the fixative in combination with a seldom-used plastic solvent. This protocol produced good ultrastructural preservation in tissues and large, high-quality, 2-micrometers thick, plastic-free sections. These semithin sections provided a level of structural and antigenic preservation, image resolution, and labeling intensity that surpassed all other conventional sectioning methods used for immunocytochemistry. The capacity to use a single tissue sample in studies designed for light and electron immunocytochemistry, in conjunction with existing autoradiographic and cytochemical techniques, makes this a very desirable method for routine tissue preparation in research and clinical applications. PMID- 6296222 TI - Postembedding immunocytochemical localization of paramyxovirus antigens by light and electron microscopy. AB - A postembedding method is described to localize antigens specific for various paramyxoviruses in sections of cells and tissues that have been fixed and embedded in epoxy resins for conventional electron microscopy. Viral antigens were localized in CV-1 cell cultures infected with simian virus 5 (SV5), brains of suckling hamsters inoculated with either neuroadapted mumps virus or hamster adapted measles virus, and brains of adult mice infected with Sendai (parainfluenza I) virus. Both 1-micrometer-thick and thin (gold) tissue sections were etched with alcoholic sodium hydroxide-solution and then treated following either the unlabeled antibody peroxidase-antiperoxidase or the biotinylated protein A:avidin peroxidase procedure. Primary reagents included immunoglobulin isolated from hyperimmune rabbit sera with specificity to the major viral components of SV5 or SV5 hemagglutinin-neuraminidase, to whole mumps virus or mumps virus nucleocapsids, and to whole Sendai virus. Crude rabbit anti-Sendai virus antiserum and whole human subacute sclerosing panencephalitis (SSPE) sera were used in parallel. The results indicate that tissues processed for conventional evaluation by electron microscopy may be suitable, within limits, for postembedding immunocytochemical staining of paramyxovirus antigens. PMID- 6296223 TI - Application of a rapid avidin--biotin--peroxidase complex (ABC) technique to the localization of pituitary hormones at the electron microscopic level. AB - The new avidin--biotin--peroxidase complex (ABC) technique was applied to ultrathin sections of rat pituitary that were fixed with glutaraldehyde and embedded in Araldite 6005. The primary antisera dilutions that are normally applied for 24-48 hr with the peroxidase-antiperoxidase (PAP) complex technique were used. High background was observed with the ABC method when incubation times were 12-48 hr. Tests were then conducted with shorter incubation times. The staining intensity was measured with a densitometer. Detectable stain was seen after only 15 min in dilutions of 1:10,000 anti-bovine luteinizing hormone (bLH beta), 1:8000 anti-rat thyroid-stimulating hormone (rTSH beta), and 1:20,000 anti 25-39-adrenocorticotropic hormone (25-39ACTH). Optimal LH staining was seen after 30 min, whereas optimal staining for TSH or ACTH required 1 hr. Stain was detectable with a dilution of 1:4000 anti-human follicle-stimulating hormone (hFSH beta) after 30 min and was optimal after 4 hr. Prolonged incubation times with these dilutions decreased the staining intensity because a deposit of high background was produced that appeared as a filigreed network over the cells. When higher dilutions were tested with 2-hr incubation times, optimal staining was seen with 1:30,000 anti-bLH beta, 1:24,000 anti-rTSH beta, 1:30,000 anti-25 39ACTH, and 1:8000 anti-hFSH beta. These tests demonstrate the potential of the ABC method for the rapid detection of small amounts of specific and nonspecific antibodies that are bound to pituitary cells. PMID- 6296224 TI - Arginine deprivation and the generation of white variants in cowpox virus infected cell cultures. AB - The white pock variant of cowpox virus shows limited growth in chick embryo fibroblasts maintained in arginine-deprived culture medium. Since these conditions inhibit the growth of parental virus, there is a marked increase in the frequency of the white variant in the virus population recovered after passage in the absence of arginine. The variants generated in this system have been characterized by restriction endonuclease analysis of virus DNA in the total DNA recovered from infected cell cultures. Such analysis shows that the white variants arise as deletion mutants of the parental virus, but there was considerable heterogeneity in the restriction patterns of different isolates examined shortly after their generation. Further passage selected white cowpox virus populations with a stable genome configuration comparable with the DNA of pock-purified white variants. PMID- 6296225 TI - A poxvirus antigen associated with pathogenicity for rabbits. AB - White pock variants of cowpox virus give papular lesions on intradermal inoculation of rabbits, without the necrosis and haemorrhage that are produced by wild type cowpox viruses. Rondle & Dumbell (1962) have shown that white pock variants of cowpox virus fail to produce a specific, precipitating antigen which they called 'd' substance. In this paper it is shown that 'd' is demonstrable in soluble antigen preparations of rabbitpox virus and of neurovirulent strains of vaccinia virus but not in soluble antigens of variola viruses. Two series of recombinant viruses prepared by Dumbell & Bedson (1964) from variola and cowpox and from variola and rabbitpox viruses were tested for the production of 'd' substance. These results were compared with the previously recorded effects of these recombinants when inoculated intradermally in rabbits. It is concluded that functional genes determining the production of 'd' and of rabbit skin pathogenicity are closely linked on the pox virus genome, but that there is insufficient evidence to say that the two functions are interdependent. PMID- 6296226 TI - The pathogenicity of variola virus. A comparison of the growth of standard strains of variola major and variola minor viruses in cell cultures from human embryos. AB - The international reference strains of variola major (Harvey) and of variola minor (Butler) were grown in cultures of skin and muscle cells from human embryos. The development of infective virus, complement-fixing antigen, haemagglutinin and cytological changes were followed at four temperatures between 35 and 40 degrees C. No significant difference was found in the amount of virus produced by Harvey or Butler viruses at any of the experimental temperatures, but Harvey attained the plateau titre at 16 h, some 4 h ahead of Butler in the cultures incubated at 38 degrees C. Harvey also produced a higher and more prolonged yield of virus in the extracellular medium of cultures, inoculated at low multiplicity and incubated at 37 degrees C. At 38 degrees C small inocula of Harvey produced foci which developed and spread till the whole culture was necrotic; Butler foci did not spread and remained relatively undeveloped at this temperature.Staining with acridine orange showed the development of cytoplasmic inclusions at all temperatures up to 39.5 degrees C, at which temperature most inclusions remained round and discrete and susceptible to digestion with DNase. The yield of virus at all temperatures correlated well with the number of cells in which the DNA cytoplasmic inclusions became irregular in outline, diffuse and insusceptible to digestion with DNase. It was concluded that elevated temperatures, up to 39.5 degrees C affected principally a maturation phase in the development of the virus.Equal amounts of complement-fixing antigen were produced at all temperatures and by either virus, but the late product, haemagglutinin was depressed at elevated temperatures much more in cultures infected with Butler than in cultures infected with Harvey. This was clearly shown by haemadsorption; in cultures, infected at high multiplicity and incubated at 39.5 degrees C Harvey gave semi-confluent haemadsorption, while only an occasional haemadsorbing cell could be found in the cultures infected with Butler virus.It is concluded that there were two ways in which temperature affected the growth of variola viruses in cultures of human embryo skin and muscle cells. The total yield of virus was reduced by inhibition of a stage in the maturation of the virus with a cut off point between 39.5 and 40 degrees C at which no cells produced infective virus, and with little observable differences between Harvey and Butler viruses. Changes at the surface of virus-infected cell, involving virus release and haemagglutinin, were affected independently of virus maturation; in these changes Butler was much more sensitive than Harvey to elevated temperatures. The relevance of these observations to the development of variola major and variola minor in man is discussed. PMID- 6296227 TI - Laboratory diagnosis of smallpox: role of the Virus Reference Laboratory, Colindale, 1947-70. AB - The Virus Reference Laboratory, Colindale, first embarked on laboratory investigations for smallpox early in 1947. From then, in conjunction with the Department of Bacteriology, University of Liverpool, it provided a complete diagnostic service throughout England and Wales until 1962, after which the service became available regionally until eradication was effected. Up to 1970 it had investigated 2696 specimens from suspected cases of smallpox and had recovered 108 strains of variola and 248 of vaccinia virus. These last were from persons suffering the complications of vaccination. Some outbreaks following smallpox importation are discussed but infection among laboratory staff during this period was not demonstrated. PMID- 6296228 TI - Electrophoretic study of the genome of human rotaviruses from Rio de Janeiro, Sao Paulo and Para, Brazil. AB - Human rotaviruses from the states of Rio de Janeiro, Sao Paulo and Para of Brazil were analysed by RNA electrophoresis. At least some bands characteristic of rotavirus double-stranded RNA were detected in 138 (86.8%) of 159 faecal samples in which the presence of rotavirus had been demonstrated by enzyme immunoassay. Of the RNA-positive samples, 18 (13.0%) were classified as subgroup 1, 94 (68.1%) as subgroup 2, and 26 (18.8%) could not be classified due to absence of visible bands 10 and 11. Subgroup 2 was more frequent in the three states. All strains of subgroup 1 detected in Rio de Janeiro were associated with a single short-lived school outbreak. All strains of subgroup 1 resembled each other in electrophoretic pattern, irrespective of geographical origin, although minor differences could be detected by co-electrophoresis. Subgroup 2, on the other hand, showed a great degree of electrophoretic heterogeneity and could be divided into several sub-categories. PMID- 6296229 TI - Hotting-up the complement-fixation test. AB - A detailed investigation into the effect of modifying the incubation temperature of the complement-fixation (CF) test is described. For varicella-zoster virus cytomegalovirus and rubella virus, increasing the incubation temperature progressively increased the sensitivity of the CF test to reach a maximum at 15 degrees C, at which temperature the geometric mean titre of seropositive samples was significantly greater than that found at 4 degrees C. For these three viruses, each serum shown to contain IgG antibodies by ultrasensitive radioimmunoassay procedures was detected by CF following incubation at 15 degrees C. No false-positive reactions occurred at 15 degrees C, but it was our impression that anticomplementary activity was enhanced at this temperature. Significant increases in antibody titre at 15 degrees C were also seen when measles virus, respiratory syncytial virus, adenovirus and Mycoplasma pneumoniae were employed as CF antigens. The results demonstrate that the CF test should be performed at 15 degrees C if optimum sensitivity is to be achieved. The ability of the test to detect significant rises in antibody titre was not impaired at the higher incubation temperature. PMID- 6296230 TI - Pteridines as a new marker to detect human T cells activated by allogeneic or modified self major histocompatibility complex (MHC) determinants. AB - Evidence is presented that activation of T cells by allogeneic or modified autologous cells leads to the specific production of two distinct molecular compounds of the PT class. One of these two PT has previously been identified as neopterin. The structure of the other is still under investigation. This conclusion was based on the following findings: a) In vitro both PT were produced by T cells proliferating in response to HLA incompatible allogeneic or to autologous EB-virus-transformed or TNP-haptenized cells. b) In vitro stimulation of T cells with mitogenic lectins or protein antigens, of macrophages with zymosan-complement, of B cells with PWM or EB-virus, and of NK effector cells with tumor targets failed to induce comparable release of these PT. c) In vivo increased amounts of both PT were excreted via the urine during allograft rejection and viral infections. It thus appears that the production of these PT is primarily a feature of activated T cells involved in the control of self integrity. PMID- 6296231 TI - Immunoglobulin secretion during the autologous mixed lymphocyte reaction in man. PMID- 6296232 TI - alpha-Interferon increases immunoglobulin production in cultured human mononuclear leukocytes. AB - Interferons (IFN) are known to modulate immune responses in either an inhibitory or a stimulating manner. The present study was initiated to investigate the mechanisms by which alpha-IFN modulates Ig production of human peripheral blood mononuclear cells (PBMC). IgG and IgM production was measured in pokeweed mitogen (PWM) stimulated 7-day cultures of PBMC. Significant enhancement of IgM and IgG production was observed when alpha-IFN was added. Overnight preincubation followed by washing also produced significant enhancement. The effect of alpha IFN was not obtained in the absence of PWM or T cells. The effect of alpha-IFN on cultures of B and T cells was not altered by irradiation of T cells (2000 rad). alpha-IFN was not shown to enhance the production of helper factor but did increase the responsiveness of B cells to helper factor if the B cells were preincubated with alpha-IFN. Finally, alpha-IFN did not increase the Ig production of PBMC induced by Epstein Barr virus (EBV), and the outgrowth of EBV infected PBMC was not affected. Overall, these results show for the first time that the effect of alpha-IFN on PBMC is due to an enhanced responsiveness of B cells to helper factors produced by radioresistant T cells. PMID- 6296233 TI - Identification of normal and transformed lymphocyte subsets of nonhuman primates with monoclonal antibodies to human lymphocytes. AB - Monoclonal antibodies (MAb) that detect specific lymphocyte subsets of man were tested for their reactivity with peripheral blood lymphocytes (PBL) and established cell lines of owl monkeys (Aotus trivirgatus) and marmosets (Saguinus spp). Results of these studies showed that certain determinants were conserved and that the pattern of reactivity observed in one genus (or even species) could not be used to predict the reactivity in another. On owl monkey PBL, reactivity equivalent to that on human PBL was observed with OKT11a, anti-Leu 3a, B1, and anti-HLA-DR MAb thus detecting T cell, helper/inducer T cell, B cell, and HLA-DR determinants. After neuraminidase treatment of owl monkey PBL, reactivity with anti-Leu 5 and OKT8 (T cell and suppressor/cytotoxic T cell determinants) was observed. Cell separation experiments indicated these determinants were found on lymphocytes with the same general properties as those of human origin. The use of these MAb to examine owl monkeys infected with Herpesvirus saimiri and tumor cell lines established from H. saimiri-inoculated monkeys revealed that the virus was found in OKT11a-positive, B1-negative cells. In chronically infected nondiseased animals, H. saimiri was found in anti-HLA-DR-negative cells and was restricted to anti-Leu 3a-positive cells (two animals) or was nearly equally distributed in anti-Leu 3a-positive and negative cells (one animal). Established H. saimiri tumor cell lines had the phenotype OKT11 a-positive, anti-HLA-DR-positive, B1 negative and were either anti-Leu 3a-positive or negative. Studies of interleukin 2-responsive tumor cells in short-term culture suggested the tumor was composed of virus-positive anti-Leu 3a-positive and negative populations. PMID- 6296234 TI - Inhibition of lymphocyte-mediated cytolysis and cyclic AMP phosphodiesterase by erythro-9-(2-hydroxy-3-nonyl)adenine. AB - The adenosine deaminase (ADA) inhibitor erythro-9-(2-hydroxy-3-nonyl)adenine (EHNA), at low concentrations (less than 10 microM), enhances the inhibitory activity of adenosine against lymphocyte-mediated cytolysis (LMC) without itself being inhibitory. At higher concentrations, EHNA alone is inhibitory to LMC with an IC50 of 160 microM. This inhibition is reversible upon washout, appears to affect an early stage of the lytic process, and does not appear to involve changes in basal levels of cyclic AMP (cAMP), ribonucleoside 5'-triphosphate pool sizes, S-adenosylhomocysteine levels, or protein carboxymethylation. EHNA does enhance the cAMP response of cytolytic lymphocytes (CL) to activators of adenylate cyclase such as prostaglandin E1. EHNA inhibits lymphocyte high affinity cAMP phosphodiesterase at immunosuppressive levels, exhibiting hyperbolic mixed-type inhibition (Ki = 83 microM, alpha = 0.47, beta = 0.18). Whereas inhibition of intralymphocytic ADA is complete at low concentrations (less than 25 microM) of EHNA, inhibition of LMC and intralymphocytic cAMP phosphodiesterase increases linearly with EHNA concentration to at least 200 microM. The presence of 200 microM EHNA during the centrifugation of mixtures of CL and EL4 leukemia target cells leads to increased CL cAMP levels. 2' Deoxycoformycin, a more potent ADA inhibitor than EHNA, is not inhibitory to LMC and shows none of these cAMP-related effects. These results suggest that CL target cell contact stimulates adenylate cyclase in the CL and that EHNA inhibits LMC due to its enhancement of this target cell-stimulated elevation of cAMP. PMID- 6296235 TI - Phosphorylation of pig epidermal soluble protein by endogenous cAMP-dependent protein kinase. AB - The distribution of adenosine 3',5'-monophosphate (cAMP)-dependent protein kinase and its substrate proteins was analyzed using soluble and particulate fractions of pig epidermal homogenates. When histone was used as a substrate for this enzyme reaction, protein kinase activity was distributed almost equally between the soluble and particulate fractions. However, the effect of exogenously added cAMP was confined almost exclusively to the soluble enzyme. Endogenous protein phosphorylation in the absence of exogenous histone was higher in the particulate fraction than in the soluble fraction, but the stimulating effect of cAMP was observed only in the soluble fraction. These results indicate that cAMP-dependent protein kinase is predominantly localized in the soluble fraction and phosphorylates soluble epidermal proteins. The particulate fraction contains protein kinase which is cAMP-independent and phosphorylates particulate-bound proteins as well as histone. Based on these observations, the soluble fraction was incubated with [gamma-32P]-ATP in the presence or absence of cAMP, and phosphorylated protein was analyzed by SDS disc- or slab-gel electrophoresis followed by autoradiography. Among many proteins whose phosphorylation was slightly increased by cAMP, a protein with Mr approximately 45,000 was found which was markedly phosphorylated in the presence of cAMP. Although this protein corresponds to one of the richest proteins in the epidermal soluble fraction, an important physiologic role for this phosphorylation has not been clarified. PMID- 6296236 TI - Cyclic AMP-dependent protein kinase isozymes of pig skin and human skin from normal and psoriatic subjects. AB - Cyclic AMP-dependent protein kinase isozymes of pig and human skin (epidermis) were separated by DEAE-cellulose column chromatography after micromodification for small biopsy samples. Clear-cut separations of type I and type II isozymes, which were of about equal amounts, could be obtained only when the ischemia effect was avoided by in vivo freezing of skin and homogenization for less than 10 s. Intradermal injections of epinephrine caused dose-dependent activation of type I isozyme, but not of type II. Injections of other skin adenylate cyclase stimulators such as histamine, adenosine, and prostaglandin E2 elevated the local cyclic AMP levels to not more than 5 pmol/mg protein and also stimulated only the type I isozyme. Incubation of keratome-sliced pig skin under various conditions caused both activation by dissociation and inactivation by reassociation of the subunits, which appeared to be dependent on the cyclic AMP content. Epinephrine added to the incubation medium led to complete activation of both type I and type II isozymes (the intraepidermal cyclic AMP contents ranged from 20-50 pmol/mg protein). The isozymes of normal skin and involved skin of psoriatics showed identical peaks of type I and type II isozymes of equal amounts. The data indicate that protein kinase in the involved skin is not in an activated (by cyclic AMP) state. PMID- 6296237 TI - Local effects of synthetic leukotrienes (LTC4, LTD4, LTE4, and LTB4) in human skin. AB - The local effects of intracutaneous injections into humans of 1-3 nmol of five products of arachidonic acid metabolism, leukotrienes (LT) C4, D4, E4, and B4 from the 5-lipoxygenase pathways and prostaglandin (PG) D2 from the cyclooxygenase pathway, were assessed clinically and histologically. In equimolar concentrations, LTC4, LTD4, and LTE4, elicited erythema and wheal formation, in which a wheal with central pallor was present up to 2 hr, and the erythema persisted as long as 6 hr. PGD2 elicited a wheal that lasted up to 1 hr and erythema that lasted up to 2 hr. The dermal vascular sites affected by LTD4 and PGD2 included capillaries, superficial and deep venules, and arterioles. LTB4 elicited a transient wheal and flare, followed in 3-4 hr by induration that was characterized by a dermal infiltrate comprised predominantly of neutrophils. The combination of LTB4 and PGD2 elicited tenderness and increased induration associated with a more intense neutrophil infiltration. Thus, the products of the 5-lipoxygenase pathway of arachidonic acid metabolism in nanomole amounts can induce cutaneous vasodilation with edema formation and a neutrophil infiltrate, and these responses are enhanced by a cyclooxygenase pathway product, PGD2. PMID- 6296238 TI - Interferon prophylaxis against simian varicella in Erythrocebus patas monkeys. AB - Erythrocebus patas monkeys were given placebo or human leukocyte interferon (5 x 10(5) units/kg of body weight per day im) for five days during an epizootic of simian varicella. During the 14 days beginning with the first day of treatment, the attack rate for simian varicella was 14.3% (two of 14) among interferon recipients compared to 70% (nine of 13) among placebo recipients (P less than 0.025). Excluding animals with antibody to simian varicella when the study began, 18% (two) of 11 interferon recipients had symptoms of infection compared to 80% (nine) of 11 placebo recipients (P less than 0.025). The epizootic began in a room housing male animals. The incidence of infection in male placebo recipients was 100% (seven of seven) compared to 14% (one of seven) in male interferon recipients (P less than 0.01). The efficacy of interferon prophylaxis in the simian varicella model supports its continued evaluation for the management of human varicella in high-risk patients. PMID- 6296239 TI - Primary and recurrent concomitant genital infection with herpes simplex virus types 1 and 2. PMID- 6296240 TI - Food poisoning due to Clostridium perfringens in the United States. PMID- 6296241 TI - Antibody response to cytomegalovirus after renal transplantation: comparison of patients with primary and recurrent infections. AB - The specific antibody response of 22 renal transplant patients with primary cytomegalo-virus (CMV) infection was compared to that of 21 patients with recurrent infection using seven different techniques to measure antibody. With primary infection seroconversion occurred between two and 12 weeks postoperatively, and geometric mean titers increased rapidly during this interval with each test except virus neutralization. In the group with recurrent CMV infection, geometric mean titers declined slightly initially and then increased rapidly to reach peak levels by 10 weeks. In both primary and recurrent infections, IgM antibody was detectable by radioimmunoassay and indirect immunofluorescence; the former procedure was clearly the more sensitive. After absorption to remove rheumatoid factor, 20 of the 22 patients with primary infection and eight of the 21 with recurrent infection had IgM antibody to CMV by radioimmunoassay which often persisted for over six months. The former group had significantly more viremia and symptomatic infections than the latter. Two critically ill patients failed to develop IgM or neutralizing antibody. PMID- 6296242 TI - Zoster in children with cancer: radioimmune precipitation profiles of sera before and after illness. AB - Sera collected from children with cancer before and for extended periods after the onset of zoster were analyzed by radioimmune precipitation techniques. The percent recovery of both [3H]fucose- and [35S]methionine-labeled varicella-zoster virus (VZV)-specific antigens increased severalfold immediately after zoster and declined slowly during convalescence; however, within two years serum panels from two patients exhibited serologic evidence of subclinical reactivation of VZV. After electrophoretic fractionation of the immunoprecipitates, the polypeptide profile after zoster closely resembled that described for high titer xenoantisera to VZV and contained at least 16 constituents ranging in molecular weight from 32 to 174,000. In contrast, sera obtained before zoster were easily distinguished because they precipitated poorly, if at all, two major VZV glycoproteins (gp62 and gp98) and several nonglycosylated polypeptides. The emergence of zoster, therefore, was associated with the appearance of previously undetectable antibodies to VZV-specific proteins. PMID- 6296244 TI - [Electron microscopic studies of human cytomegalovirus.--characterization of intracellular forms of parental viral DNA molecules before and after onset of the viral DNA synthesis]. PMID- 6296243 TI - Oral administration of human rotavirus to volunteers: induction of illness and correlates of resistance. AB - Four of 18 volunteers challenged orally with human rotavirus strain D (subgroup 2, serotype Wa) developed a diarrheal illness two to four days after inoculation. Viral shedding was detected in five of the 18 volunteers, whereas 12 (67%) developed serologic evidence of infection. Two volunteers who developed diarrheal illness after the initial inoculation were given the same inoculum 19 months later; neither developed diarrhea, although one developed constitutional and gastrointestinal symptoms. The presence of preinoculation serum immunofluorescent antibody to rotavirus strain D or high levels of neutralizing antibody to Wa or reassortant DS-1 human rotavirus correlated with resistance to diarrheal illness. Although prechallenge serum antibody correlated with resistance to diarrhea and/or shedding of rotavirus, the relationship of preexisting local neutralizing activity in intestinal fluid was less clear-cut. PMID- 6296245 TI - [Normal inhibitor(s) to the hemagglutinin of human polyomaviruses, BK virus and JC virus]. PMID- 6296246 TI - Cyclic AMP dependent protein kinase in human placenta. AB - There have been few reports on cyclic AMP dependent protein kinase in the human placenta. As our first study on this enzyme we here report the confirmation of its existence in human placental chorion and its chemical characteristics and the measured activities of the enzyme in each gestational trimester. 1. Chemical characteristics: (1) pH Profile: Maximum specific activity was found to be between pH7.5 and 8.5. (2) Effects of Mg2+: Magnesium ions activated protein kinase activity. Maximum specific activity was found to be 5mM. (3) Km for ATP: Km was found to be 4.94 microM. 2. Activity of cyclic AMP dependent protein kinase of human placental chorion in each gestational trimester: The specific activities (p moles ATP/mg protein/minute) of each gestational trimester were 44.67 +/- 5.29 for the first, 45.24 +/- 9.13 for the second and 34.19 +/- 4.95 for the third trimester. There was a significantly lower value in the third trimester (t less than 0.001), compared to the first and second trimester. PMID- 6296247 TI - [Therapeutic effects of dimethyl sulfoxide on the urinary proteins of patients with familial amyloidotic polyneuropathy]. PMID- 6296248 TI - Bilateral Wilms' tumor. A case report and review of the literature. PMID- 6296249 TI - Expression and excretion of human fibroblast beta 1 interferon in monkey cells after transfection with a recombinant SV40 plasmid vector. AB - We have constructed a eukaryotic expression vector designed to express a gene under late SV40 transcriptional control. From this chimeric plasmic-SV40 vector, virtually all the sequences which code for the major capsid protein VP1 have been deleted and instead, the human fibroblast interferon beta 1 cDNA gene has been inserted. After transfection of monkey cells with this recombinant, substantial quantities of human beta 1 interferon (up to 2 x 10(-4) IU/ml) were excreted in the culture medium. Transfection of nonpermissive mouse L cells or rat cells yielded virtually undetectable quantities of human beta 1 interferon (5 x 10(3) to 10(4) times less than that in monkey cells). The recombinant SV40 vector may serve as a model vehicle for the efficient expression of other eukaryotic genes and might also be used as a direct screening vector for cloning of eukaryotic or prokaryotic cDNA genes. PMID- 6296250 TI - Viable polyoma virus with four BamHI decanucleotide linkers inserted at the 26 minute HincII site. AB - BamHI decanucleotide linkers (5' CCGGATCCGG 3') were ligated to full-length linear polyoma DNA partially cleaved with HincII, and the recombinant DNA was transfected into mouse 3T6 cells. A viable virus (PYNB5) was isolated which contains four linkers at the 26-min HincII site. PYNB5 is encapsidated with a larger major viral capsid protein (VP1) as predicted from the DNA sequence. PYNB5 appears to have the same growth and physical properties as the wild-type virus with the exception of greater inactivation upon dilution of virus stocks with water. When PYNB5 DNA is cleaved with BamHI, the larger of the two resulting fragments (PY66) contains an intact early region of the virus, including the origin of DNA replication. PY66 complements a polyoma early region mutant for growth and may be useful as a cloning vector for foreign DNA. PMID- 6296251 TI - ISRm1: A Rhizobium meliloti insertion sequence that transposes preferentially into nitrogen fixation genes. AB - After transposon Tn5 mutagenesis, a high proportion of Rhizobium meliloti symbiotic mutants do not contain Tn5 insertions in symbiotic genes. Instead, the mutations in these strains are correlated with the presence of an endogenous insertion sequence (ISRm1) in nitrogen fixation (nif) or symbiotic genes which are adjacent to the nif genes. ISRm1 is 1.4 kb and transposes to at least three restriction fragments in the nif region at a frequency between 10(-2) and 10(-3). A nif region restriction fragment containing ISRm1 was cloned from one of the mutant strains unable to fix nitrogen symbiotically (Fix-) and the resulting plasmid was used as a hybridization probe. ISRm1 is present at least ten times in the R. meliloti genome but is not present in any other R. meliloti strains, E. coli strains, or Rhizobium species tested. We demonstrated that the Fix- phenotype correlated with ISRm1 transposition is indeed caused by ISRm1 insertion by conjugating a cloned fragment containing ISRm1 into a wild type Fix+ R. meliloti host and replacing the normal genomic nif fragment with the nif::ISRm1 fragment. The resulting strain was Fix-. PMID- 6296252 TI - Construction of adenovirus expression vectors by site-directed in vivo recombination. AB - We developed a method for conveniently positioning foreign DNA at many preselected sites in the adenoviral genome by a combination of in vitro and in vivo recombination. Using this technique, we constructed a set of recombinant viruses that contain the SV40 A gene downstream from the adenovirus tripartite leader. One of these hybrid viruses, Ad-SVR26, contains the A gene close to and downstream from both the major late promoter and the first segment of the tripartite leader. The transcripts encoded by the inserted SV40 DNA are highly overproduced in infected cells; they initiate at the adenoviral late promoter and terminate at the SV40 polyadenylation site. Several novel splice acceptor sites in the SV40 sequences are used in the processing of the primary transcript to produce six different species of spliced RNA. The synthesis of T antigen in Ad SVR26-infected cells requires the use of novel AUG initiation codons present within the SV40 coding region or adenoviral sequences that normally form part of the intron between the first and second segments of the tripartite leader. The level of T antigen expression is not as high as the level of mRNA production. The usage of these new AUG triplets or the absence of the complete adenovirus tripartite leader sequence may account for the low efficiency of translation. PMID- 6296253 TI - Deletion mapping of DNA regions required for SV40 early region promoter function in vivo. AB - The SV40 early region promoter, previously localized to the DNA segment bounded by the HpaII and HindIII restriction sites (nucleotides 346 and 5171), was further defined by construction of an extensive set of deletions within this region and measurement of their effects on (a) viral DNA replication, (b) virus multiplication and the ability to complement early and late mutations, (c) transformation of rat cells, (d) large T antigen formation, and (e) the location of the 5' ends of early mRNAs. One set of mutations is represented by deletions that begin at the HpaII site and extend unidirectionally for varying lengths toward the BglI site at ori. A second set of mutants contains deletions that start at ori and extend unidirectionally for varying lengths towards the HpaII site. A third set of mutants, with deletions or duplications of various lengths and boundaries, lie between the HpaII and BglI sites. Our studies indicate the following. (a) Ori, the sequence needed for initiating SV40 DNA replication, extends from the sequences needed for initiating SV40 DNA replication, extends from the sequences needed to bind T antigen to the palindrome in site II to nucleotide 34, the late region edge of the AT block. Flanking sequences adjacent to the AT block facilitate DNA replication. (b) The SV40 early region promoter comprises two functionally distinct nucleotide sequence elements. One is flanked by nucleotides 5231 and 107, and contains the RNA initiation sites at nucleotides 5231-5237, a positioning element resembling the TATAAATA consensus sequence about 20-25 nucleotides upstream, and an RNA polymerase II recognition sequence contributed by short GC-rich sequences clustered between nucleotides 35 and 107; we refer to this as the RNA polymerase II interaction site. The second distinct sequence element is contained within each of two 72-bp segments located between nucleotides 107 and 250; the behavior of this element suggests that it may influence the accessibility of RNA polymerase II for the interaction site or the efficiency of RNA chain initiation. Large T antigen binding sites I, II, and III overlap with the putative RNA polymerase II interaction site; since large T antigen does not prevent elongation of RNA transcripts initiated upstream, T antigen probably represses early region expression by preventing RNA polymerase II binding to the promoter. PMID- 6296254 TI - A fragment of Dictyostelium discoideum genomic DNA that complements the URA1 mutation of Saccharomyces cerivisiae. AB - A strain of Saccharomyces cerivisiae carrying a mutation in the URA1 gene was transformed with fragments of Dictyostelium DNA inserted into a plasmid capable of replication in E. coli or yeast. Rare prototrophic colonies were recovered that all contained the parent plasmid and an insert of Dictyostelium DNA. Resistance to the antibiotic G418, a function coded by the same plasmid, was also expressed in the prototrophs. Plasmids recovered from the prototrophic yeast could be used to transform E. coli. The E. coli transformants harbored plasmids capable of transforming yeast URA1 mutants to prototrophy. No complementation of the E. coli pyrD mutation, which corresponds to URA1, occurred. Southern blot analysis revealed that the insert of Dictyostelium DNA contained a unique sequence of 1700 base pairs and a repetitive one of 1000 base pairs. Subcloning experiments showed that only the unique sequence was required for complementation, which is independent of the orientation of the Dictyostelium sequence in the plasmid. The repetitive fragment was not linked to the unique sequence in the genome and was probably an artifact of the ligation procedure. The unique sequence hybridized to a Dictyostelium polyA+ RNA species of 1200-1300 bases. PMID- 6296255 TI - A plasmid that replicates in both mouse and E. coli cells. AB - Cloned bovine papilloma virus (BPV) DNA induces cellular transformation when introduced into mouse cells growing in culture; the transferred viral DNA replicates as an extrachromosomal, closed circular element. BPV DNA is therefore an inviting replicon to construct a "shuttle" vector that can replicate in both mammalian cells and E. coli. Although BPV DNA devoid of bacterial plasmid sequences has been successfully employed to reintroduce cloned genes into rodent cells, construction of a true shuttle plasmid has been hampered by a disruptive influence of bacterial plasmid sequences that appear to block cellular transformation when included on the transferred molecule. We constructed a molecule, pGP, containing the transforming region of the BPV genome, the rat growth hormone gene, and bacterial plasmid pBR 327, and have found unexpectedly that the intact molecule can induce cellular transformation of mouse cells at high efficiency despite the presence of bacterial sequences in the transferred plasmid. A similar plasmid without the growth hormone segment does not transform mouse cells. The pGP molecule replicates as a stable plasmid in both mouse cells, where there are 30 to 80 monomer episomes per cell, and E. coli, and may be shuttled back and forth unaltered between the two kinds of cells. The growth hormone gene is transcribed in the mouse cells and gives rise to a transcript that is longer than authentic rat growth hormone mRNA and does not appear to be regulated by glucocorticoids. When pGP is cotransferred into mouse L-cells with herpes simplex virus tk gene, it appears to integrate, and free monomer episomes are not observed. PMID- 6296256 TI - Splicing of intervening sequences introduced into an infectious retroviral vector. AB - The v-src gene was removed from Rous sarcoma virus DNA and replaced with either a cDNA or genomic clone of human alpha-chorionic gonadotropin. Transfection of the recombinant retrovirus genomes into normal chicken fibroblasts produced nontransforming recombinant virus in high titer. Neither helper virus nor selective conditions were needed. Cells infected with the recombinant viruses expressed RNA containing the gonadotropin sequences in levels equivalent to those of term human placenta (approximately 0.5% of poly A+). Essentially every fibroblast in the culture was infected; the infected cells contained approximately one recombinant provirus each. The gonadotropin intervening sequences were removed precisely from the recombinant genomes that contained them, creating recombinants carrying a perfect cDNA copy of the original genomic insert. However, the intervening sequences were removed inefficiently such that several viral replicative cycles were necessary before all genomes had been processed completely. The implications of these observations to the transduction of viral oncogenes and the creation of processed pseudogenes are discussed. PMID- 6296257 TI - Symbiotic nitrogen fixation: molecular cloning of Rhizobium genes involved in exopolysaccharide synthesis and effective nodulation. AB - A transposon (Tn5)-induced mutant (strain ANU437) of Rhizobium trifolii was isolated in which no water-soluble exopolysaccharide (EPS) could be detected. This mutant was also incapable of forming nitrogen-fixing root nodules on clover plants. Molecular cloning has demonstrated that the Tn5 transposon was responsible for both of these mutant phenotypes and that there is a direct correlation between EPS synthesis in this bacterial strain and its ability to carry out symbiotic nitrogen fixation. In the mutant ANU437, Tn5 was located in a 9.4-kb EcoRI fragment that was cloned into the amplifiable plasmid pBR322. The recombinant plasmid was used as a hybridization probe to isolate the corresponding wild-type DNA sequence of R. trifolii from a lambda Charon 28 genomic clone bank. This DNA sequence was subcloned into the broad host range conjugative plasmid RP4 and introduced into the Escherichia coli strain RR1. It was then transferred to the mutant ANU437 by conjugation. The acquisition of the wild-type DNA sequence by the mutant ANU437 resulted in the restoration of its ability to synthesize normal levels of EPS and to form nitrogen-fixing nodules on white and subterranean clovers. PMID- 6296258 TI - Transport of free and conjugated steroids from the boar testis in lymph, venous blood and rete testis fluid. AB - In 12 anaesthetized boars the concentrations of oestrone sulphate and dehydroepiandrosterone sulphate (DHAS) were 15- to 35-fold higher in lymph collected from a vessel in the spermatic cord than in testicular venous blood plasma from a vein in the spermatic cord. The concentrations of testosterone, total unconjugated oestrogens and dehydroepiandrosterone (DHA) were about twofold higher in lymph. The concentrations of all steroids studied were higher in testicular venous blood plasma than in arterial blood plasma (testosterone about sixfold; total unconjugated oestrogens about fourfold; oestrone sulphate about threefold; DHA and DHAS about twofold), but the concentrations of testosterone, total unconjugated oestrogens and oestrone sulphate in rete testis fluid were comparable to those in arterial blood plasma. Lymph flow from the pig testis was about 7% of plasma flow so that about 80% of the oestrone sulphate and DHAS produced by the testis leaves the organ in the lymph; the comparable values for testosterone, total unconjugated oestrogen and DHA were about 20%. In the 90-min period following an injection of human chorionic gonadotrophin there were substantial increases in the concentration of testosterone and smaller increases in the other steroids in arterial and spermatic venous blood plasma and in testicular lymph, but not in rete testis fluid; there were also small increases in lymph flow, but no change in blood flow. PMID- 6296259 TI - Accumulation of thiourylene antithyroid drugs in mouse salivary gland. AB - [35S]Methimazole and [35S]propylthiouracil were shown to accumulate in mouse submandibular gland in vivo, with maximal tissue: plasma ratios being achieved at the lowest dose of drug studied (0.1 microgram/animal). Autoradiography of submandibular glands showed that the drugs were localized to the intralobular ductal epithelium and within the lumen of the convoluted granular tubule, which was identical to the localization of radiolabelled iodide. Histochemical studies indicated that this was the site of peroxidase activity within the gland. Drug accumulation persisted when iodide trapping was competitively inhibited using perchlorate. These data suggest that antithyroid drug accumulation by this tissue is not dependent on the anion trap; the localization of drug and iodide at the site of peroxidase activity suggest that this may be an important factor in the mechanism of drug accumulation, possibly related to subsequent drug metabolism. PMID- 6296260 TI - A method for obtaining chimaeric mouse blastocysts with two separate inner cell masses: a preliminary report. AB - Pairs of zona-free mouse blastocysts aggregated in the presence of inactivated Sendai virus and subsequently cultured in vitro will fuse to form a chimaeric blastocyst with one common blastocoelic cavity. Depending on the relative position of the inner cell masses in the apposed 'parental' blastocysts, the resulting chimaeric blastocyst contains either a single inner cell mass (ICM) of dual origin or two discrete ICMs each originating from one embryo. In the present experiments, fusion between the two aggregated blastocysts occurred in 23% of the pairs and 64% of these chimaeric blastocysts contained two ICMs. Blastocysts of the latter type could potentially give rise to pairs of embryos which as regards the topography of the foetal membrane would resemble spontaneous identical twins, although they would be genetically dissimilar. Possible applications of the described method are discussed. PMID- 6296261 TI - Correlation between blastocyst oxygen consumption and trophoblast cytochrome oxidase reaction at initiation of implantation of delayed mouse blastocysts. PMID- 6296262 TI - The iron-H2O2-iodide cytotoxic system. AB - A potent antimicrobial system is described which consists of ferrous sulfate (Fe2+), hydrogen peroxide (H2O2), and iodide in 0.02 M sodium acetate buffer pH 5.5. H2O2 could be replaced by the H2O2-generating system glucose + glucose oxidase. This system, unlike the myeloperoxidase-H2O2-halide system, was ineffective when iodide was replaced by bromide, chloride, or thyroxine, and was inhibited by EDTA, the hydroxyl radical scavengers mannitol and ethanol, and phosphate and lactate buffers at the same concentration and pH as the acetate buffer used. The acetate buffer, however, could be replaced by water. It is proposed that Fe2+ and H2O2 (Fenton's reagent) generate OH X (or a closely related substance), which interacts with iodide to form one or more toxic species. PMID- 6296263 TI - Simultaneous flow cytometric analysis of human T cell activation antigen expression and DNA content. AB - Cell-surface antigens that are induced to appear on T cells activated by the lectin phytohemagglutinin-P (PHA) can be classified both on the basis of the kinetics of their appearance and on their growth-association properties. Seven distinct T cell activation antigens, defined by monoclonal antibodies, were classified as early, intermediate, or late antigens based on their temporal appearance relative to DNA synthesis. Four antigens, the transferrin receptor, the T cell activation antigen Tac, the 4F2 antigen, and the 49.9 antigen were early antigens, whereas the OKT10 antigen appeared at intermediate times and both HLA-DR and antigen 19.2 appeared late. The use of a dye, Hoechst 33342, which stains DNA stoichiometrically, allowed the simultaneous analysis of immunofluorescence and cell cycle position of individual cells. This analysis unexpectedly revealed that essentially all cells in the proliferative phase of the cell cycle expressed each of the four early-activation antigens. The correlation between expression of the four early-activation antigens and T cell proliferation suggests that these molecules are important for the growth of all T cells. The relationship of two of these activation antigens, known to be the receptors for transferrin and interleukin 2, a T cell growth factor, is discussed with special reference to the roles of their ligands in supporting the growth of T cells. PMID- 6296264 TI - Experimental autoimmune thyroiditis. In vitro cytotoxic effects of T lymphocytes on thyroid monolayers. AB - Effector mechanisms in experimental autoimmune thyroiditis (EAT) were studied in vitro by establishing a cytotoxicity system with thyroid target cells. Lymph node cells (LNC) from popliteal and inguinal lymph nodes were obtained from CBA/J mice (8-10 wk old) 12-18 d after immunization with 120 micrograms mouse thyroglobulin (MTg) in complete Freund's adjuvant (0.2 ml to both hind footpads and thighs) and were cultured with MTg (10-50 micrograms/ml). On day 5 of culture, viable LNC were added to labeled thyroid monolayers and their cytoxicity was assayed after 16 h. Functional thyroid target cells, as reflected by MTg production for up to 9 d, were prepared by adding 1 mM dibutyryl adenosine 3',5'-cyclic monophosphate and 60 microU thyroid-stimulating hormone/ml to the culture medium. On days 5-7, confluent monolayers were labeled with 111In and used as targets. Specific 111In release ranged from 56 to 85%. The cytotoxic response is MTg specific and H-2 restricted. Pretreatment of thyroid target cells with rabbit antiserum to MTg completely inhibited cytotoxicity. Pretreatment with mouse antiserum to either Kk or Dk products resulted in approximately 50% inhibition, whereas the combined use of both antisera led to total inhibition. No cytotoxicity was observed when control BALB/c thyroid cultures were the target cells. The kinetics of the expansion of Thy-1+ cytotoxic cells by in vitro exposure to MTg were then studied. The cytotoxic response required 5 d to develop and was abolished by treating LNC on day 4 with monoclonal antibody to Lyt-1.1, but not to Lyt-2.1, plus complement. In contrast, by day 5, cytotoxicity was abrogated by similar treatment with antiserum to Lyt-2.1, but not to Lyt-1.1. We conclude that cytotoxic cells derived from MTg-immunized mice are Lyt-2-bearing cells but require the presence of Lyt-1-bearing cells for their generation and/or differentiation. PMID- 6296265 TI - Specific binding of leukotriene B4 to a receptor on human polymorphonuclear leukocytes. AB - In this paper we have described the binding of nanomoler concentrations of [3H]leukotriene B4 (LTB4) to human polymorphonuclear leukocytes. Because up to 80% of the total [3H]LTB4 binding was blocked by excess (greater than 100 times) [14C]LTB4, the majority of binding is specific. Stereospecificity of the LTB4 binding is demonstrated by the diminished relative abilities of the 6-trans-and 12-epi-6-trans- isomers of LTB4 to block [3H]LTB4 binding. With these two isomers 3-10-fold higher than [14C]LTB4 concentrations were needed for equivalent inhibition of [3H]LTB4 binding. This difference is quantitatively less dramatic than the differences between these isomers in many in vitro functional assays such as chemokinesis, chemotaxis, and degranulation. Binding of [3H]FMLP is not blocked at greater than 100-fold excess of LTB4. The binding of [3H]LTB4 to cells appears to be essentially irreversible at 4 degrees C, but not at 37 degrees C where initially bound LTB4 is rapidly converted to metabolites which then enter the medium. These results suggest the presence of a saturable, stereospecific site for LTB4 on PMN. The association of LTB4 binding and the initiation of pharmacological responses to LTB4 will require further studies. PMID- 6296266 TI - Interleukin 1 stimulation of collagenase production by cultured fibroblasts. AB - Interleukin 1 is a monokine that exerts biological effects on a variety of target cells in vitro. In this report, interleukin 1 has been found to be capable of stimulating collagenase production by cultured dermal fibroblasts. The concentrations of interleukin 1 that stimulate fibroblast collagenase production are similar to those that stimulate mouse thymocyte proliferation. Analyses by high performance liquid chromatography indicate that interleukin 1, rather than a contaminating monokine, is responsible for this effect on fibroblasts. Interleukin 1, released in vivo by macrophages infiltrating sites of tissue damage or inflammation, may function to stimulate the release of collagenase by connective tissue fibroblasts. PMID- 6296267 TI - Binding of carcinoembryonic antigen (CEA) to an anti-CEA immunosorbent in the presence of detergents. AB - The binding of [125I]CEA to an anti-CEA Sepharose-4B immunosorbent has been investigated in the presence of detergents. Non-ionic detergents (Triton-X-100, Lubrol PX and Torgitol NPX) at concentrations of 5-40 milligrams did not inhibit the binding of [125I]CEA to the immunosorbent. Sodium deoxycholate decreased the binding from 98% (concentration of detergent 0.6 milligram) to 41% (concentration of detergent 20 milligrams). Sodium dodecyl sulfate inhibited the binding of [125I]CEA to the immunosorbent considerably, even at low concentrations. PMID- 6296268 TI - Developmental aspects of hexose metabolism in Bufo bufo. AB - Due to the close correlation between glucose mobilization and utilization within animal tissues, in this paper, the stages of appearance of phosphorylase, glucose 6-phosphatase and hexokinase as well as the levels of some intermediates of glucose metabolism have been investigated during Bufo bufo development. Phosphorylase first appears at stage 13 and is dominant in the neural part of the embryo, but, after this stage, increases relatively more in the nonneural one. Hexokinase appears at stage 17 and glucose-6-phosphatase soon after. Phosphorylase appearance at stage 13 is correlated with an increase of lactate content in the embryo; this may indicate a metabolization of hexoses. On this basis, the subsequent appearance of hexokinase and glucose-6-phosphatase activities also seems coherent with hexose mobilization and utilization within embryo. No direct causative factor for the changes observed was evident. PMID- 6296269 TI - Cyclic nucleotides during chondrogenesis: concentration and distribution in vivo and in vitro. AB - This study correlates endogenous levels of cAMP and cGMP with their immunohistochemical localization during chondrogenic differentiation of C57B1/6J mouse limb mesenchyme in vivo and in vitro. A transient decrease in cGMP but not cAMP was found from days 12 to 13 in vivo correlating with early stages of chondrogenesis in the developing limb. Intracellular levels of both cAMP and cGMP in high density limb mesenchyme cultures increased 25% after 24 hr in culture when aggregate and nodule formation was detectable. When cells were seeded at different initial plating densities to delay the onset of aggregate and nodule formation, increased levels of intracellular cAMP correlated temporally with the appearance of nodules. Both cyclic AMP and cGMP were immunohistochemically localized in perichondrial cells and chondrocytes in vivo and in vitro. Therefore, (1) cAMP levels correlated temporally with the appearance of chondrogenic cells and (2) cAMP and cGMP were immunohistochemically localized to chondrogenic cells. These data indicate that fluctuations of both cAMP and cGMP levels may be involved in limb cartilage differentiation. Although increases in both nucleotides were found to correlate with the onset of chondrogenesis in vitro, in vivo data suggest that the amount of cAMP relative to cGMP rather than the absolute amount of an individual cyclic nucleotide may be more significant in modulating differentiation. PMID- 6296270 TI - Primary tumors of the liver in infancy and childhood. PMID- 6296271 TI - Distribution of the Escherichia coli K12 insertion sequences IS1, IS2 and IS3 among other bacterial species. PMID- 6296272 TI - Enzymes of N-acetylglucosamine metabolism during germ-tube formation in Candida albicans. AB - The enzymes of N-acetyl-D-glucosamine (GlcNAc) metabolism, GlcNAc-6-phosphate deacetylase and GlcN-6-phosphate deaminase were found to be inducible in Candida albicans. The pattern of induction for these enzymes was the same under conditions of germ-tube formation (37 degrees C) and where yeast cells metabolized GlcNAc with no change in morphology (28 degrees C); this indicates that these enzymes are not control points in the dimorphic development of C. albicans. During induction there was a 40-and 25-fold increase in specific activity for the deacetylase and the deaminase, respectively, and the maximum specific activity correspond to the time when all the GlcNAc had been metabolized. The presence of lomofungin (an inhibitor of transcription) or trichodermin (an inhibitor of translation) in cell suspensions of C. albicans containing GlcNAc prevented the increase in specific activity of these enzymes. 2 Deoxyglucose inhibited germ-tube formation, partially inhibited the induction of the deacetylase (43%) and the deaminase (60%), but did not affect the growth of C. albicans on either Glc or GlcNAc. GlcN-6-phosphate was a competitive inhibitor of the deacetylase with a Ki of 1.4 mM while the other product of the reaction, acetate, did not inhibit the enzyme. The Km value for GlcN-6-phosphate on GlcN-6 phosphate deaminase was 0.24 mM. Incubation of starved yeast cells with GlcNAc produced a four-fold increase in the specific activity of UDP-GlcNAc pyrophosphorylase at either 28 degrees C or 37 degrees C. PMID- 6296273 TI - Distribution of cytochromes in selected species of corynebacteria pathogenic to animals. PMID- 6296274 TI - Inhibition of nicotinic acid and nicotinamide uptake into Bordetella pertussis by structural analogues. AB - 3-Pyridine-carboxaldehyde and 3-pyridine-aldoxime were effective and specific inhibitors of the uptake of both nicotinic acid (NA) and nicotinamide (ND) by Bordetella pertussis, although neither compound inhibited the growth of the bacteria in liquid medium or the oxidation of glutamate by washed suspensions. In contrast, the following pyridine derivatives did not inhibit uptake of NA or ND: iso-NA, iso-ND, isoniazid, 6-amino-NA and 6-amino-ND, 3-acetyl-pyridine, 3 pyridyl-acetic acid, N,N-diethyl-ND and 3-pyridine-sulphonic acid. 3- Pyridyl carbinol was inhibitory, but less so than the first listed compounds. PMID- 6296275 TI - Isolation and characterization of a recombinant plasmid carrying a functional part of the Bacillus subtilis spoIIA locus. AB - Plasmid pHM2 consists of a 3.3 kb insert of Bacillus subtilis DNA in the chimeric plasmid pHV33, and can replicate in Escherichia coli and B. subtilis. In B. subtilis, pHM2 complements the defects resulting from mutations spo-42, spo-50, spo-69 and sas-1 in the spoIIA locus. This complementation can occur in recE4 strains where recombination of the plasmid with the chromosome is prevented, and the chromosome retains the mutant allele. Thus the plasmid carries a functional part of the spoIIA locus; it does not contain the complete locus as it cannot complement several other spoIIA mutations. It is likely that the locus is complex, containing at least two genes. PMID- 6296276 TI - Molecular cloning of bovine papillomavirus genomes and comparison of their sequence homologies by heteroduplex mapping. AB - The genomic DNAs of bovine papillomavirus (BPV) type 1, type 2 and type 4 were cloned in pAT153. BPV1 and BPV2 genomes were cloned using the single HindIII sites of the vector and virus DNAs, and BPV4 was cloned using the single BamHI sites. The orientation of the recombinant DNAs was established by restriction enzyme digestion, hybridization and heteroduplex analysis. The results showed that: (i) BPV1 and BPV2 DNAs are in register and are broadly homologous throughout most of their length when aligned at their single HindIII sites; (ii) depending on the degree of hybridization stringency used, the two DNAs show one major region and several minor regions of partial homology, mainly residing in the segment of the genomes believed to contain the structural genes; (iii) BPV4 DNA shares no homology with either BPV1 or BPV2 DNA. PMID- 6296277 TI - Murine coronaviruses: isolation and characterization of two plaque morphology variants of the JHM neurotropic strain. AB - Two plaque-size variants of the neurotropic JHM strain of mouse hepatitis virus have been isolated from the virus stock after eight serial passages in suckling mouse brain. One variant, JHM-DL, produces large plaques, while the other, JHM DS, produces small plaques in tissue culture. DS replicates more slowly, has a lower virus yield in vitro, and is less virulent for mice than DL. They also differ in their pathogenicity for mice: JHM-DL infection results in acute encephalomyelitis while JHM-DS infection results in demyelination. Oligonucleotide fingerprint analysis of the RNA genomes of these two variants revealed that they had almost identical genetic sequences. Each variant, however, had a unique oligonucleotide spot not found in the other. The unique spot of the large plaque variant, JHM-DL, was localized at approximately 3 to 5 kb from the 3' end, while the JHM-DS unique spot was mapped at 14 to 15 kb from the 3' end of the genome. We have further shown that these oligonucleotide changes are not correlated with the plaque morphology. These two viruses may be useful for studying the molecular basis of virus-induced demyelination. PMID- 6296278 TI - Thymidine kinase deletion mutants of herpes simplex virus type 1. AB - Deletions in the cloned thymidine kinase (TK) gene of herpes simplex virus type 1 (HSV-1), strain 17 syn+, were produced by two methods. Removal of a 506 base pair fragment from between the unique SstI and Bg/II restriction endonuclease sites of pTK1 (HSV-1 BamHI p cloned in pAT153) and subsequent transformation of Escherichia coli resulted in the isolation of 50 deleted plasmids. Sequential digestion of pTK1 with Bg/II and nuclease BAL 31 followed by ligation and recleavage with Bg/II resulted in the isolation of 31 deleted plasmids. Three clones, pTK2, pTK3 and pTK4, obtained following Bg/II and SstI treatment of pTK1 were recombined with wild-type (wt) HSV-1 (17) syn+ DNA in baby hamster kidney (BHK) cells to produce TK- deletion mutants HSV-1 (17) TK 1301, HSV-1 (17) TK 1302 and HSV-1 (17) TK 1303 respectively. 5-Bromo-2'-deoxyuridine, 5-bromo-2' deoxycytidine and 9-(2-hydroxyethoxymethyl)guanine were used to reduce the background of TK+ virus in heterogeneous recombinant stocks analysed for the presence of TK- recombinants. All recombinant clones isolated produced a small syncytial plaque morphology in BHK cells. The mutants HSV-1 (17) TK 1301 and HSV 1 (17) TK 1302 were TK-, failed to produce polypeptides of molecular weights 43000 and 19000 found in wt-infected cells and demonstrated one-step growth curves different from wt virus and the TK- mutant HSV-1 (17) dPyk-7. Superinfection studies with HSV-1 (17) TK 1301, HSV-1 (17) TK 1302, HSV-1 (MDK) and HSV-1 (17) dPyk-7 indicated that all TK- mutants except dPyK-7 produce a trans-acting gene product which can switch on the transforming HSV-1 TK gene. PMID- 6296279 TI - Monoclonal antibodies to three non-glycosylated antigens of herpes simplex virus type 2. PMID- 6296280 TI - Low infectivity of HSV-1 DNA caused by defective-interfering genomes. AB - The infectivity of herpes simplex virus, type 1, strain ANG progeny DNA from standard virus infections and of progeny DNA from infections involving defective interfering virus particles (DI DNA) was compared in transfection assays. No difference in infectivity of virus DNA isolated either from infected cells or from progeny virus was found for a given type of infection. However, the values for two types of infection differed markedly, with DI progeny DNA being less infectious by more than 2 log10. The low infectivity was mainly due to the presence of interfering DNA molecules in DI progeny DNA, regardless of whether intracellular DNA or DNA extracted from mature virions was analysed. The interfering capacity of DI progeny DNA did not depend on the integrity of the genomes. The physical proximity provided by simultaneous precipitation of infectious and of interfering DNA is an important factor influencing the degree to which DI DNA interferes. Interference by DI DNA in the transfection assay can be partly reversed by the addition of XbaI fragments of standard DNA; in control experiments this fragmented DNA was shown to lead to a reduction rather than to an enhancement of the infectivity of standard virus DNA. PMID- 6296281 TI - Characterization of non-oncogenic Marek's disease virus-infected and turkey herpesvirus-infected lymphocytes. PMID- 6296282 TI - Adsorption and penetration of enveloped herpes simplex virus particles modified by tunicamycin or 2-deoxy-D-glucose. PMID- 6296283 TI - Enhancement of plasminogen activator activity by the culture medium of Rous sarcoma virus-transformed cells. AB - We have tested the effect of the culture medium of chicken embryo fibroblasts (CEF) transformed by Rous sarcoma virus (RSV) on plasminogen activator (PA) activity of normal and RSV-infected cells. The results obtained showed that fibrin digestion by cultures of normal uninfected and RSV-infected cells directly attached to a fibrin substrate was increased when the cells were exposed to the supernatant of an 18 h-culture of CEF transformed by RSV. Addition of cycloheximide to the supernatant completely abolished this effect. Similarly, exposure of both normal and RSV-infected cells to the culture medium of RSV transformed CEF resulted in a marked increase of the soluble PA activity present in the supernatant. This effect, however, was not abolished by inhibition of protein synthesis. For both the cell-bound and soluble PA activities of normal and RSV-infected cells the stimulatory effect was transient, being maximal between 8 and 12 h of exposure to the transformed cell-conditioned medium and disappearing after 24 h. A possible correlation with transformation-enhancing factor(s), previously reported to be present in the culture medium of transformed cells, is discussed. PMID- 6296284 TI - Nature of the antibody response to the foot-and-mouth disease virus particle, its 12S protein subunit and the isolated immunizing polypeptide VP1. AB - Inoculation of inactivated 146S foot-and-mouth disease virus particles into guinea-pigs elicited the formation of neutralizing antibody and the serum had a 10-fold higher titre in radioimmunoassay (RIA) with 146S particles than with the 12S virus subunit. In contrast, a single inoculation of the 12S subunit or the isolated polypeptide VP1 elicited the formation of antibody having a much lower titre in RIA with the 146S particle than with the 12S subunit and low or undetectable neutralizing activity. However, sera from guinea-pigs given two or more inoculations of the 12S subunit or VP1 had neutralizing activity. The level in the anti-VP1 serum was lower than that in the anti-12S serum and both were much lower than that in animals receiving two inoculations of the 146S particle. The neutralizing activity elicited by the three antigens was absorbed by the homologous antigen. In contrast, neither the 12S subunit nor VP1 absorbed the anti-146S neutralizing antibody and VP1 did not absorb the anti-12S subunit neutralizing antibody. However, the 12S subunit partly absorbed the neutralizing activity elicited by VP1. The results are compatible with a model in which the 146S particle elicits a spectrum of neutralizing antibodies which are completely absorbed by the homologous particle but only partially by the 12S subunit or VP1. The results are discussed in relation to the structural features required for the production of neutralizing antibody. PMID- 6296285 TI - African swine fever virus DNA: restriction endonuclease cleavage patterns of wild type, Vero cell-adapted and plaque-purified virus. AB - DNA from African swine fever (ASF) virus was isolated and was characterized by two restriction enzymes, SmaI and EcoRI. Although both enzymes can distinguish Vero cell-adapted ASF isolates by characteristic restriction endonuclease cleavage patterns, all ASF isolates examined exhibited a high degree of similarity, as measured by co-migration of most of the DNA fragments. The molecular weight of ASF DNA, based on size estimates of DNA fragments from cleavage patterns, ranged from 93 x 10(6) to 100 x 10(6). Virus genome heterogeneity was observed in uncloned, cell culture-adapted ASF isolates as well as in a plaque-purified virus after serial passage in Vero cells. In contrast to the rather minor differences in restriction pattern among the Vero cell-adapted isolates, a major alteration in restriction endonuclease cleavage sites was observed during adaptation of the wild-type virus to cell culture. PMID- 6296286 TI - Transformation of human embryonic fibroblasts by BK virus, BK virus DNA and a subgenomic BK virus DNA fragment. AB - Human embryonic fibroblasts (HEF) have been transformed by BK virus (BKV) DNA and by u.v.-inactivated or live BKV alone or in association with methyl-cholanthrene (MTC). The transformed cells produced BKV large T and small t antigens as well as the cellular 53 kdal protein, detected by immunofluorescence and immunoprecipitation. After an initial phase of lysis and virus shedding, virus or its coat protein antigen could not be detected in transformed cells. All human transformed cell lines could be superinfected by BKV or BKV DNA, but their susceptibility to superinfection was 20- to 500-fold lower than normal HEF. BKV could be rescued by fusion of transformed cells with normal HEF or Vero cells and by transfection of normal HEF with total DNA and DNA extracted from the Hirt supernatant of transformed cells. Blot hybridization analysis of DNA from transformed cells showed a considerable amount of free BKV DNA in monomeric and polymeric forms. Integrated BKV DNA was absent in most cell lines but present in only small amounts in BKV-transformed cells treated with MTC. Analysis of free BKV DNA with various restriction endonucleases and by blot hybridization showed that monomeric forms were complete BKV genomes, whereas polymers contained both complete and defective or rearranged BKV DNA. Transformation of HEF was also obtained with a 3.7 kilobase (kb) fragment of the BKV genome, produced by sequential digestion of BKV with the restriction endonucleases HhaI and EcoRI. This fragment extends clockwise on the virus genome from 0 to 72.2 map units and contains the entire early region. Blot hybridization analysis of cells transformed by the HhaI/EcoRI 3.7 kb fragment showed two separate integrations of BKV sequences without free virus DNA. PMID- 6296287 TI - Organization of polyoma virus DNA in mammary tumours of athymic mice. AB - Restriction mapping of polyoma virus DNA in mammary tumours of athymic mice gave patterns that varied with the tumour examined. These reflected differences in both the organization and the state of integration of virus genomes in the host chromosomes. All tumours contained tandemly integrated full-length and defective virus genomes. Some tumours also contained unintegrated virus DNA molecules, some full-length and others defective. The deletions were localized in the virus genomic sequences coding for the distal part of the large T antigen. After the first transplantation, the organization of polyoma virus genomes in tumours remained essentially unchanged through four successive transplantations. The tumour cells that initially contained free virus DNA molecules continued to possess such molecules during serial transplantations. The virus DNA molecules in transplanted tumours lacking unintegrated virus genomes were more methylated than in tumours containing unintegrated virus genomes. PMID- 6296288 TI - Localization of rotavirus antigens in infected cells by ultrastructural immunocytochemistry. AB - Virus structural antigens were localized within a line of monkey kidney (MA104) cells infected with the simian rotavirus SA11 using electron microscopic immunoperoxidase techniques. When hyperimmune guinea-pig anti-SA11 serum was used, virus particles, membranes of virus-associated endoplasmic reticulum, and viroplasmic inclusions were most heavily labelled. A general cytoplasmic reaction (ribosomes, intracytoplasmic membranes, etc.) with anti-SA11 serum was also observed, but nuclei were unstained. In addition, several other virus-induced structures were found to contain rotavirus proteins, including convoluted smooth membrane within the endoplasmic reticulum, aberrant virus-like particles, and 15 to 20 nm diam. cytoplasmic tubules. Monospecific antiserum to VP7 (outer capsid glycoprotein, mol. wt. 38000) reacted strongly with virus particles and the virus associated endoplasmic reticulum, but reacted poorly with viroplasmic inclusions. The nucleus and general cytoplasm were unstained with anti-VP7. In contrast, monospecific antisera to VP2 and VP6 (inner capsid proteins, mol. wt. 94000 and 41000 respectively) reacted very strongly with viroplasmic inclusions. Virus particles, endoplasmic reticulum and cytoplasmic ribosomes were also labelled with these sera. These results indicate that rotavirus inner capsid proteins are synthesized throughout the cytoplasm and become concentrated in viroplasmic inclusions, while the outer capsid glycoprotein is synthesized primarily on ribosomes of the rough endoplasmic reticulum. Thus, the outer capsid layer appears to be acquired during virus budding into cisternae of the endoplasmic reticulum. PMID- 6296289 TI - Dissemination of herpes simplex virus in nude mice after intracutaneous inoculation and effect of antibody on the course of infection. AB - Dissemination of herpes simplex virus (HSV) in nude mice after intracutaneous inoculation in the midflank, and the effect of passively administered antibody on the course of infection were investigated. In untreated infected mice the skin lesions developed rapidly and HSV could first be recovered from the homogenate of the dorsal root ganglia on day 3 after infection, from the spinal cord on day 7 and from the brain on day 11. HSV could not be recovered from the blood, spleen or liver. In mice passively immunized with human gamma globulin, development of the skin lesions was rather slow and HSV could not be recovered from the homogenate of the dorsal root ganglia until day 16. From the results of explant culture of the ganglia, HSV was found to have reached the ganglia as early as 48 h after infection, even in mice administered human gamma globulin. The protective action of antibody seems to originate from the inhibition of virus growth not only at the inoculation site but also in the dorsal root ganglia. PMID- 6296290 TI - Identification of the envelope surface glycoproteins of equine herpesvirus type 1. AB - The structural polypeptides of purified enveloped virions of the Army 183 strain of equine herpesvirus type 1 (EHV-1) were examined by different analytical techniques to identify the envelope glycoproteins. Glycoproteins were identified by electrophoretic analysis in polyacrylamide slab gels of virus labelled in vivo with [3H]glucosamine or labelled enzymically in vitro with either UDP [14C]galactose or sodium [3H]borohydride. Fluorograms revealed eleven glycoproteins (mol. wt. 260000, 150000, 138000, 90000, 87000, 65000, 62000, 60000, 50000, 46000, and 24000). These glycoproteins probably correspond to virion protein (VP) 1-2, 9b, 10, 13, 14, 16, 17, 18, 21, 22a and 25 respectively, as designated in two other EHV-1 strains. In addition, a poorly resolved glucosamine-rich region (mol. wt. 250000 to 200000) corresponded to VP 3 to 8. The two isotopic surface labelling methods revealed that all the virus glycoproteins were exposed on the envelope surface. PMID- 6296291 TI - Vesicular stomatitis virus can establish persistent infections in Syrian hamsters. AB - Persistent infections by vesicular stomatitis virus (VSV) of the Indiana serotype were readily established in adult Syrian hamsters following intraperitoneal injection of the virus. Plaque-forming virus, identified as VSV by serological and physical criteria, was isolated from brain homogenates of five hamsters that were tested 3 to 8 months after infection. Four of these animals had exhibited either transient or permanent paralysis, whereas the fifth appeared healthy, during the period of observation. At the time of sacrifice all hamsters had high titres of anti-VSV-neutralizing antibodies in their sera. PMID- 6296292 TI - A viable simian virus 40 variant with a deletion in the overlapping genes for virion proteins VP1, VP2 and VP3. AB - Nucleotide sequence analysis was used to determine the exact location of a deletion in the late region of the SP2 mutant of simian virus 40 (SV40), a viable small-plaque variant isolated from a persistent infection of rhesus monkey kidney cells. The results indicate that six base pairs are deleted from that part of the SV40 genome in which the coding regions for the three virion proteins, VP1, VP2 and VP3, overlap. This implies that all three virion proteins are affected by the deletion. This finding is discussed with respect to the viability of SP2. PMID- 6296293 TI - Recombinational joints in a simian virus 40 variant generated in a persistent infection. AB - SP1, a viable simian virus 40 (SV40) variant isolated from a persistent infection of rhesus monkey kidney cells, contains sequence rearrangements in the untranslated region of the SV40 genome which are transcribed into late mRNA leader sequences and in the region which encodes the large T antigen. Nucleotide sequences about the recombinational junctions in SP1 were determined. The sequence data show that in most instances there was not extensive homology between recombining sequences. The recombinant sequences are discussed with respect to the mechanisms by which they might have been generated. PMID- 6296294 TI - Inversion of the two segments of the herpes simplex virus genome in intertypic recombinants. AB - We have analysed by restriction site mapping the structures of the termini and L S joint in several HSV-1/HSV-2 intertypic recombinants, including Bx1(28-1), the virion DNA of which has a marked overabundance of one orientation of the L segment, and subclones of Bx1(28-1). All recombinants with both orientations of L present in equal amounts contain TRL and IRL regions derived at least in part from the same parent (HSV-1 or HSV-2) as a result of previously undetected crossovers in these regions. Recombinants with a predominance of one orientation of L have TRL and IRL regions derived from different parents. Homology between a sequences alone at the L terminus and L-S joint is sufficient for normal inversion of L. Analysis of another recombinant, RE4, which fails to invert normally in both L and S, suggests that normal inversion of S is dependent upon the presence of TRS and IRS regions derived at least in part from the same parent. We conclude that segment inversion specifically depends upon the a sequence, that the process of DNA replication and maturation does not necessarily produce molecules with identical a sequences, and that direct ligation of termini may occur during DNA replication. PMID- 6296295 TI - RNA-dependent RNA polymerase activity in murine coronavirus-infected cells. AB - The multiplication of murine coronavirus strains A59 or JHM in Sac(-) cells was unaffected by the presence of alpha-amanitin at concentrations which inhibited the host cell DNA-dependent RNA polymerase activity. In cells infected with the A59 virus strain, actinomycin D-resistant RNA synthesis could readily be detected by pulse-labelling with [3H]uridine; this virus-specific RNA synthesis was not induced in the presence of the protein synthesis inhibitor anisomycin. A new RNA dependent RNA polymerase activity was detected in the large particle fraction of A59 virus-infected cells. Optimal conditions for enzyme activity in vitro were established. Maximum activity occurred 5 h after infection, coincident with the peak of virus-specific RNA synthesis detected by pulse-labelling in vivo. PMID- 6296296 TI - Isolation of influenza C virus from pigs and experimental infection of pigs with influenza C virus. PMID- 6296297 TI - Proteins specified by herpesvirus saimiri: identification and properties of virus specific polypeptides in productively infected cells. AB - The synthesis and accumulation of more than 30 virus-induced polypeptides was detected after infection of permissive primate cells with high multiplicities of herpesvirus saimiri. These virus-induced polypeptides had apparent mol. wt. of from 12,000 (12K) to 250K and differed in molar abundance by up to two orders of magnitude. The majority of virus-induced polypeptides satisfied multiple criteria for their virus specificity. Polypeptides of 110K, 76K, 51K and 31K to 29K were synthesized at maximum rates early in infection, but the majority of proteins were made at high rates late in infection. Virus-specific polypeptides were substrates for a number of post-synthetic modifications. The 117K, 85K, 76K, 31K and 30K polypeptides were each processed to forms with altered electrophoretic mobility. The 117K and 85K polypeptides were among the virus-specified substrates for glycosylation and virus-induced polypeptides of 59K, 51K, 30K and 26K were phosphorylated. The early 51K phosphoprotein and the 30K to 31K polypeptides were rapidly translocated to the nucleus of infected cells. The 31K polypeptide was processed to a 29K product which remained stably associated with the nuclear fraction. The 30K nuclear protein was shown to be the precursor of a 28K polypeptide which was released in a soluble form into the cytoplasmic fraction and the culture medium of cells at late times in the virus growth cycle. Many other polypeptides accumulated slowly in nuclear (e.g. 250K, 150K, 130K, 110K and 38K) or in cytoplasmic (e.g. 117K, 85K and 28K) fractions of infected cells in forms which could be differentiated by the use of detergents or differences in stability to salt extraction. The mol. wt., relative molarities and some features of the post-synthetic processing of herpesvirus saimiri polypeptides more closely resembled the properties of gene products of Epstein-Barr virus than those of herpes simplex virus. PMID- 6296298 TI - Involvement of glycolipids in myxovirus-induced membrane fusion (haemolysis). AB - Myxoviruses (influenza viruses and paramyxoviruses) penetrate their host cells by fusion of viral and cellular membranes. During this process, virus envelopes react with galactose-terminating glycolipids of cellular membranes. This was suggested by experiments showing the ability of these lipids to inhibit fusion (as measured by haemolysis) when reacted with the viruses, and to enhance it when added as enrichment to erythrocytes. PMID- 6296299 TI - Differential expression of endogenous virus glycoprotein in fibroblasts and sera of some adult chickens. AB - A rapid and convenient assay for the expression of endogenous retrovirus glycoprotein in adult chickens has been developed based on the enzyme-linked immunosorbent assay (ELISA) principle. This method has been standardized using the conventional chick helper factor test. In the course of establishing this method with a large number of specific pathogen-free (VALO) chickens, an interesting diversity became apparent; about 20% of the birds which, according to chick helper factor tests performed with feather follicle fibroblast cultures were negative for endogenous virus glycoprotein expression, exhibited relatively high titres of reactive glycoprotein in serum. However, in no case was a chick helper factor-positive animal negative in serological tests. The possibility of endogenous virus antigen expression which either cannot be detected in fibroblasts, or is incapable of functioning in the chick helper factor complementation, is discussed. PMID- 6296300 TI - Alterations in coxsackievirus B4 heart muscle disease in ICR Swiss mice by anti thymocyte serum. AB - Coxsackievirus B4 (CB4) infection in infant ICR Swiss mice induces synchronous peaks in both virus titres and pathologic changes in the heart. Among surviving mice, transmural necrosis is followed by fibrosis and ventricular aneurysm. Rabbit antimouse thymocyte serum (ATS) was given before CB4 infection to determine the effects of thymus-dependent functions upon the course of disease. The mortality in ATS-treated mice was 75.9% (65 of 83 mice died) compared to 21.3% (16 of 75 mice died) in normal rabbit serum-treated controls. Pathologic studies showed that ATS-treated mice had more extensive myofibre necrosis and subsequent mineralization, but during the first 10 days of infection, leukocytic infiltration was decreased. Splenic follicles were not present until the 17th day after infection in this treated group. Serum CB4-neutralizing antibodies were similar in mice from the group treated with ATS and normal rabbit serum. These findings indicate that ATS-suppressible functions contribute importantly to virus elimination, perhaps by an increase in macrophage phagocytosis of CB4. PMID- 6296301 TI - Isolation and characterization of anti-rotavirus immunoglobulins secreted by cloned hybridoma cell lines. AB - Five monoclonal hybridoma cell lines secreting antibodies against bovine rotavirus have been produced and four of them characterized by immunostaining of structural polypeptides electrophoretically transferred on to nitrocellulose sheets. Three hybridomas appeared to be directed against the major structural polypeptide (VP39) of the virion. These three monoclonals cross-reacted with the major polypeptide of simian rotavirus and human rotavirus. A fourth hybridoma appeared to react specifically with the high-molecular weight external polypeptide (VP89) and its cleavage products. A cross-reaction was observed with human Wa strain but not with SA11. The fifth hybridoma, even though reacting in an immunofluorescent test, did not show any reactivity by immunostaining. None of the monoclonals neutralized the infectivity of bovine rotavirus. PMID- 6296302 TI - Disaggregation and reconstitution of oligomeric complexes of simian virus 40 large T-antigen. AB - Biochemical properties of multiple species of simian virus 40 (SV40) large T antigen produced in SV40-infected monkey cells were investigated by zonal sedimentation centrifugation of radiolabelled cell extracts on sucrose gradients. Two major subpopulations of T-antigen detected by immunoprecipitation and gel electrophoresis could be distinctly separated: low molecular weight forms ranging approximately between 5S and 10S and higher oligomeric forms at about 16S to 23S. Removal of divalent cations by chelating agents such as EDTA disassembled higher oligomers into low molecular weight forms (5S to 10S). Adding divalent cations in excess of the EDTA concentration reassembled the higher oligomeric forms, showing a sedimentation behaviour like T-antigen in untreated cell extracts. Our results are compatible with the hypothesis that divalent cation binding properties of T antigen participate in the natural pathway of assembling multiple oligomeric species. PMID- 6296303 TI - Either orientation of the L segment of the herpes simplex virus genome may participate in the production of viable intertypic recombinants. AB - We have analysed the genome structures of 90 recombinant viruses produced by co infection of baby hamster kidney cells with the DNA of a herpes simplex virus type 1 temperature-sensitive mutant (tsD) and specific DNA fragments of wild-type herpes simplex virus type 2 at non-permissive temperature. Crossovers were located predominantly in regions of greatest intertypic homology, and we conclude that primary recombination occurred with the L segment of the genome in either orientation. PMID- 6296304 TI - Herpesvirus saimiri-induced proteins in lytically infected cells. I. Time-ordered synthesis. AB - The addition of TPA (phorbol-12-myristate-13-acetate) to cultures during the lytic infection with herpesvirus saimiri led to an enhanced and accelerated production of polypeptides induced by H. saimiri and to a rapid shut-down of host cell protein synthesis and allowed a detailed analysis of the protein patterns. Analysis of sequential protein synthesis in owl monkey kidney cells lytically infected with H. saimiri 11 permitted the identification of 31 virus-induced polypeptides. The use of the amino acid analogues canavanine (for arginine) and azetidine (for proline) in parallel allowed experiments on the identification of proteins synthesized early and late during lytic infection. PMID- 6296305 TI - Comparison of the physical maps of the DNAs of two cytomegalovirus strains. AB - The physical maps of the DNAs of two cytomegalovirus isolates, AD169 and SG, were compared by cross-blot hybridization and by hybridization of nitrocellulose-bound SG fragments with cloned 32P-labelled AD 169 fragments. From this comparison it can be concluded that both physical maps are co-linear to a large extent. Most variability existed at the termini of the long and short component (at the repeats). Other differences were the presence or absence of restriction endonuclease cleavage sites. PMID- 6296306 TI - A paralytic disease in nude mice associated with polyoma virus infection. AB - Nude mice (nu/nu), heterotransplanted with human tumours and kept in isolators, were found to suffer from wasting and posterior paralysis. Electron microscopy of spinal cord tissue revealed virus particles in the oligodendrocytes consistent in size (35 to 40 nm), morphology and distribution with those of the polyoma--SV40 sub-group of papovaviruses. Serology and restriction enzyme analysis of the virus genome showed that the virus was the murine polyoma A2 strain. Inoculation of uninfected nude mice with 10(7) TCID50 of polyoma A2 strain virus produced a similar disease in these mice with wasting and, after 10 to 23 weeks, paralysis of the hind legs of all surviving mice. Extensive myelin disruption was seen throughout the brain stem and sacral region of the spinal cord and high titres of polyoma virus were found in the whole brain (10(8.8) TCID50/brain) and in the spinal cord (10(6.8) TCID50/spinal cord). PMID- 6296307 TI - Integration of polyoma virus DNA into chromosomal DNA in transformed rat cells causes deletion of flanking cell sequences. AB - In order to find out whether polyoma virus (Py) integration into chromosomes causes rearrangements in the cell DNA flanking the integration site, we have mapped the flanking sequences in the inducible LPT line of Py-transformed rat cells and the corresponding sequences in normal rat fibroblasts, and then compared the two maps. To carry out this study we have cloned a segment including Py DNA and flanking sequences in the bacteriophage vector lambda gt WES and subcloned the flanking cell DNA in a bacterial plasmid. We performed a Southern blot analysis of LPT and rat fibroblast DNA digested with various restriction enzymes and used the cloned flanking cell DNA and Py DNA as hybridization probes. Autoradiography of the LPT DNA blots revealed two sets of fragments. One set includes fragments containing both Py and cell DNA sequences; the second set consists of fragments which contain no virus DNA sequences, and are identical to the fragments observed in the corresponding normal rat DNA digests. These data indicate that LPT cells are heterozygous with respect to the Py inserts. The same data were used to map the flanking sequences in the two types of cells. A comparison between the two maps revealed that a 3.0 kb cell DNA segment, which is located next to the unoccupied integration site in the normal rat chromosomes, has been deleted from the LPT chromosome which carries Py DNA, but not from the LPT chromosome which does not carry the virus DNA. The implications for papovavirus integration are discussed. PMID- 6296308 TI - Virus gene expression in rat cells transformed by avian myelocytomatosis virus strain MC29 and avian erythroblastosis virus. AB - Virus gene expression in rat cells transformed by either avian myelocytomatosis virus strain MC29 or avian erythroblastosis virus has been studied by biological and biochemical methods. In the clones examined, virus-specific sequences were found to be transcribed into RNA and, in most clones, the characteristic gag related proteins could be identified. The transformed rat cells were fused to permissive chick cells and the rescued virus was shown to transform both chick embryo fibroblasts and the appropriate haemopoietic cell type in chick bone marrow cultures. These results clearly demonstrate that, as with the non defective avian sarcoma viruses, the genetic information responsible for transformation by the defective avian leukaemia viruses can be expressed in non permissive mammalian host cells as well as in permissive avian cells. PMID- 6296309 TI - Separation of a murine leukaemia virus protein kinase activity from its Pr65gag polyprotein substrate after DNA--cellulose chromatography. AB - We have recently found that, in vitro, the murine leukaemia virus (MuLV) associated protein kinase activity predominantly phosphorylates Pr65gag, a virus protein present in relatively small amounts in partially purified virus preparations. Other virus proteins, such as p10, Pr27gag and Pr40gag, are also phosphorylated in vitro, but to a lesser degree. Furthermore, when immature core subparticles which are enriched in Pr65gag are prepared from virions by Sepharose 6B exclusion column chromatography, about 50% of the kinase activity (as assayed with the exogenous substrate phosvitin) remains associated with the cores. We report here that this core-associated activity is distinct from Pr65gag since it can be separated from Pr65gag by chromatography on denatured DNA--cellulose columns followed by centrifugation of the 0.2 M-NaCl-eluted fraction. Under these conditions, Pr65gag is pelleted while the kinase activity, which can phosphorylate both endogenous (MuLV Pr65gag and p10) as well as exogenous (phosvitin) substrates, remains in the supernatant. Interestingly, when the amount of Pr65gag is reduced, as in such preparations, p10 then becomes more heavily phosphorylated. PMID- 6296310 TI - False RIA IgM titres to herpes simplex virus and cytomegalovirus: factors causing them, and their absorption by protein A-Sepharose/IgG-protein A-Sepharose. AB - A method for the absorption of false radioimmunoassay (RIA) IgM titres against herpes simplex virus (HSV) and cytomegalovirus (CMV) is presented. The serum specimens were absorbed by a mixture of protein A-Sepharose and protein A Sepharose saturated with normal human gamma globulin (PAS/IgG). The detection of rheumatoid factor of IgM class (IgM-RF) as well as antinuclear antibodies (ANA) of both IgM and IgG class by solid-phase RIA is also described, and their role in the false IgM results was studied. It was found that the PAS/IgG absorption removed 50-90% of both IgM-RF and total IgG. The reduction of IgM-ANA clustered at 50-90% or nothing, whereas the reduction of IgG-ANA was approximately 50%. The studies with HSV and CMV antigens indicated that the removal of false IgM titres was more effective than the removal of each of these four factors. It was concluded that the IgM-RF titres alone were not sufficiently high to explain the false IgM results, but the ANA activity probably contributed. PMID- 6296311 TI - Variance in rotavirus infection rates in different urban population groups in South Africa. AB - Rotavirus infection in black infants contrasts markedly with that of white infants in being much less common and showing no seasonal variation. In this multicentre study in Johannesburg, the aetiology of winter infantile gastroenteritis in black, coloured, and white infants was investigated. Stools were examined by electron microscopy and also by enzyme-immunoassay to detect subparticular antigen which may be missed by electron microscopy in patients presenting late in the course of the illness. Stools were also examined bacteriologically by conventional techniques. Rotavirus was the most common pathogen in all three population groups with bacteria playing a relatively minor role. Striking differences were observed in the rotavirus rates between the three groups. Infection in the whites was five times more common than in the blacks (60% versus 12%) with the coloureds intermediate at 40%. The hypothesis was put forward that the relative protection of the black population may be due to a greater degree of colonization of neonates, thus inducing protection against symptomatic infection at the target age of 6 to 24 months. This may well have important implications in immunoprophylaxis. The reason for the lack of seasonal variation in the black population is still unclear. PMID- 6296312 TI - Prevalence of antibodies to human cytomegalovirus in urban, kibbutz, and Bedouin children in southern Israel. AB - Prevalence of antibody to human cytomegalovirus (CMV) was determined in a sample of 860 healthy children aged 1-13 years in Beer Sheva, in the Negev region of Israel. Three groups of children were tested: (1) urban Jewish children of low, middle, and high socioeconomic levels; (2) 8 rural communes (kibbutz) in which children live in close contact with each other from the 6th week of life under good hygienic conditions and a high standard of living; (3) Bedouin, seminomadic Arabs living in relatively poor hygienic conditions in the desert. Kibbutz children showed a significantly higher rate of CMV seropositivity by the second year of life than urban Jewish and Bedouin children (76% versus 44% and 54% respectively) with a gradual increase to 94% in the 10-13-year age group. Among children living in urban populations a significantly higher prevalence of CMV seropositivity was associated with crowding, but not with other socioeconomic indicators (place of residence, country of origin, or education level of parents). A marked rise of CMV seropositivity with age was found in urban Jewish children in the 2-5-year age group during which time they attend nurseries (44% to 67%) and in the Bedouin children in the 6-9-year age group (59% to 86%) when they first attend school. The data suggest that close contact is of major importance in CMV infection in childhood. The clinical implications of early acquisition and high prevalence of CMV antibodies in the kibbutz setting are discussed. PMID- 6296313 TI - Specific IgM class antibody production following infection with cytomegalovirus. AB - Specific IgM class antibody production was studied in different groups of patients with characterized cytomegalovirus (CMV) infections using a radioimmunoassay (RIA). In pregnant women, IgM antibodies were detected only following primary infection and generally persisted less than 4 months. The demonstration of CMV-specific IgM during pregnancy is therefore diagnostic of recent primary CMV infection. In patients with symptomatic CMV infections, the appearance of IgM antibody was shown to be closely related to the onset of symptoms and coincided with production of complement fixing (CF) antibody. IgM antibodies were at maximum levels 3-4 weeks after presentation but generally declined to low or undetectable levels by 3-4 months. The significance of the results of testing for CMV-specific IgM in relation to clinical and other serological findings in these patients is discussed. IgM antibody production was also demonstrated in renal transplant patients with primary infections and in 6 of 21 recipients with secondary infections. In both groups the antibodies became detectable 3-6 weeks after transplantation but the titres were much higher following primary infection. IgM antibodies persisted throughout follow-up periods of up to 2 years after transplantation in some cases. PMID- 6296314 TI - Detection of parainfluenza IgM antibody by hemadsorption immunosorbent technique. AB - A hemadsorption immunosorbent technique (HIT) was developed for the detection of immunoglobulin M (IgM) to parainfluenza virus types 1, 2, and 3. Twenty-six (90%) of twenty-nine patients under 6 yr of age from whom parainfluenza virus was isolated showed parainfluenza IgM antibody in one or both of their paired sera, with titres ranging from 320 to 81,920. In about one third of the cases IgM antibody was demonstrated in the initial sera taken 1 to 3 days after the onset of illness. Heterotypic IgM antibody responses were observed in about 20% of the patients. The HIT test was more sensitive than the hemagglutination inhibition (HI) and complement fixation tests in detecting a seroresponse in the 29 virus positive children. The results of studies in older patients with HI titre rises to parainfluenza virus suggested that reinfection probably induced IgM antibody in a proportion of cases. The HIT test proved to be specific for the IgM class of antibody and avoided false-positive results due to rheumatoid factor. It permits an early presumptive diagnosis in a proportion of patients with parainfluenza infection. PMID- 6296315 TI - A case report of post-transfusion hepatitis A. PMID- 6296316 TI - Regulation of the muscarinic acetylcholine receptor: effects of phosphorylating conditions on agonist and antagonist binding. AB - Incubation of rat brain synaptic membranes under phosphorylating conditions (i.e., in the presence of Mg2+, ATP, and cyclic AMP) leads to a loss in muscarinic acetylcholine receptors, detectable as specific binding of the muscarinic antagonist L-[3H]quinuclidinyl benzilate. A role for protein phosphorylation in this receptor loss is indicated by the finding that 5' adenylyl imidodiphosphate, a nonhydrolysable analogue of ATP, does not support receptor loss. Furthermore, receptor loss is inhibited by adenosine and 2 deoxyadenosine, both of which inhibit protein kinase activity. The loss of muscarinic receptors is calmodulin dependent, and it has been demonstrated here that this requirement is probably at the level of calmodulin-dependent phosphorylation. An investigation of the effects of phosphorylation on the binding of the agonist carbachol to synaptic membranes from the cortex and cerebellum demonstrated that phosphorylation altered the relative proportions of the super-high-, high-, and low-affinity binding sites. The results were consistent with an apparent conversion of high- into super-high-affinity sites. In the presence of 5'-guanylyl imidodiphosphate, agonist binding demonstrated the properties expected of a population of largely low-affinity sites. This conversion of super-high- and high-affinity sites into low-affinity sites by 5' guanylyl imidodiphosphate was partially inhibited by phosphorylation. PMID- 6296317 TI - High- and low-affinity binding of [3H]imipramine in mouse cerebral cortex. AB - Binding of [3H]imipramine in mouse cerebral cortex was found to be nonhomogeneous. Competition experiments, Scatchard analysis, and Hill plots are compatible with the existence of binding with high (nanomolar) and low (micromolar) affinity. Low-affinity binding could be eliminated by the use of low concentrations of imipramine as the competing ligand. In contrast to the high affinity binding, the low-affinity binding was found to be unrelated to the neuronal uptake system for serotonin. PMID- 6296318 TI - Corticotropin peptides and melanotropins elevate the level of adenosine 3':5' cyclic monophosphate in cultured murine brain cells. AB - Cell cultures derived from mouse and rat brain and consisting mainly of astroblasts are known to respond to several hormones by increasing or decreasing their intracellular concentration of cyclic AMP. In the present study these cultures were analyzed for their susceptibility to various additional hormonal and other neuroactive compounds. Only the peptides of the corticotropin (ACTH)/melanotropin (MSH) family were found active. Their potency for elevating the intracellular level of cyclic AMP decreases in the sequence (values for the half-maximally stimulating concentrations, EC50, in parentheses) ACTH-(1-24) (10 m) greater than alpha-,beta-MSH (30 nm) greater than ACTH (greater than or equal to 100 nm) gamma-MSH, ACTH-(1-10), -(4-10), -(4-11) (greater than or equal to 0.5 microM). The lack of additivity of the maximal effects of the peptides suggests that they all act at the same receptor. The stimulation exerted by these peptides is partially suppressed by hormones known to inhibit cyclic AMP formation in that culture, i.e., noradrenaline (acting via an alpha-adrenergic receptor), adenosine (acting via an A1 receptor), and somatostatin. It is concluded that the receptors for the ACTH/MSH peptides and the inhibitory hormones are located on the same cells, presumably the astroblasts. The maximal response to ACTH and alpha- and beta-MSH depends strongly on the age of culture. The results are discussed in view of the facts that (1) peptides of the ACTH/MSH family affect behavior and learning in animals, and (2) ACTH and alpha-MSH occur in brain. PMID- 6296319 TI - Evidence for the presence of GTP-dependent regulatory component of adenylate cyclase in myelin from rat brain. AB - A GTP-dependent regulatory component of adenylate cyclase was found in myelin from rat brain. The fraction solubilized from myelin contained a component that reconstituted guanine nucleotide-responsive adenylate cyclase activity when combined with the catalytic unit of adenylate cyclase prepared from rat brain. Purified myelin demonstrated little adenylate cyclase activity, even in the presence of F- or Mn2+. The reconstituted activity was dependent on the amount of the solubilized myelin fraction and required the presence of 5' guanylylimidodiphosphate, a hydrolysis-resistant analog of GTP. The elution pattern of the component solubilized from myelin in gel filtration was very similar to that of a GTP-dependent regulatory component from synaptic plasma membranes. The content of the regulatory component-like activity in myelin was estimated to be 50-60% of that in synaptic plasma membranes. Cholera toxin ADP ribosylated proteins having molecular weights of 48,000, 38,000, 23,000, 20,000, and 15,000 and other minor peptides in myelin, some of which were also present in synaptic plasma membranes. We conclude that myelin contained a GTP-dependent regulatory component of adenylate cyclase despite the apparent lack of adenylate cyclase activity in myelin. PMID- 6296320 TI - Induction of cytosolic progestin binding sites by catecholestrogens in rat pituitary gland and uterus: different potencies of 2- and 4-hydroxyestradiol. AB - The ability of catecholestrogens to induce cytosolic progestin binding sites in the hypothalamus, pituitary gland, and uterus of ovariectomised-adrenalectomised rats was demonstrated by the increase in high-affinity [3H]promegestone binding sites (KD 1.39, 0.50, and 0.54 nM, respectively) following a single subcutaneous injection (26.4 micrograms/animal) of the 3.4-dibenzoate ester of 4 hydroxyestradiol. The affinity and the time course of induction of these binding sites were very similar to those after a single injection of an equivalent dose (20 micrograms/animal) of estradiol 3-benzoate, exhibiting maximal receptor levels after 44 h. Widely differing efficacies in the induction of progestin binding sites were observed between the dibenzoate esters of 2- and 4 hydroxyestradiol. 2-Hydroxyestradiol 2,3-dibenzoate was ineffective in the pituitary gland up to a dose of 132 micrograms/animal, whereas 4-hydroxyestradiol dibenzoate was equipotent to estradiol benzoate, showing a maximal induction of progestin binding sites at single doses in the range of 13.2-26.4 micrograms/animal (equivalent to 10-20 micrograms of estradiol benzoate). As compared to the pituitary gland, the uterus was much more sensitive to the systemic administration of estrogen benzoates. At single doses in the range of 1.32-6.6 micrograms/animal (equivalent to 1-5 micrograms of estradiol benzoate), 4-hydroxyestradiol dibenzoate induced maximal levels of progestin receptors, and even 2-hydroxyestradiol dibenzoate, when given at a high dose (132.4 micrograms/animal, equivalent to 100 micrograms of estradiol benzoate), produced a slight increase in progestin binding sites. PMID- 6296321 TI - Cyclic nucleotide-dependent protein kinases and some major substrates in the rat cerebellum after neonatal X-irradiation. AB - The levels of cAMP-dependent protein kinase (type I), or cGMP-dependent protein kinase, or protein I, and of a 23,000 MW substrate for the cGMP-dependent protein kinase were measured in cerebella from normal rats and in the cerebella from rats in which a selective loss of interneurons in the cerebellar cortex had been produced by X-irradiation. A decrease was observed in the concentrations of cAMP dependent protein kinase and of protein I, whereas an increase was observed in the concentrations of cGMP-dependent protein kinase and of the 23,000 MW substrate. The data, taken together with the results of other studies, support the interpretation that cAMP-dependent protein kinase and protein I are distributed throughout the cerebellum, but that cGMP-dependent protein kinase and the 23,000 MW substrate are highly concentrated in Purkinje cells. PMID- 6296322 TI - Noradrenalin-inducible cyclic-AMP accumulation in rat cerebral cortex: changes during complete global ischemia. AB - Neurologic dysfunction after cerebral ischemic insults may be due not only to neuronal death, but also to a possibly reversible failure in synaptic transmission. Because noradrenaline (NA)-inducible cyclic-AMP (cAMP) accumulation in brain may reflect the integrity of synaptic transmission mechanisms and brain viability, we studied its changes in cerebral cortex after various durations of decapitation ischemia. Unanesthetized rats were decapitated and the brains were kept at 37 degrees C for times ranging from 0 to 60 min. Cerebral cortical slices were incubated in vitro and NA (11.2 microM)-induced cAMP accumulation was evaluated over 10 min. At 0 min of ischemia, NA-induced cAMP accumulation was 56 pmol/mg protein/10 min. Between 0 and 20 min of ischemia, a linear eightfold increase, to 435 +/- 49 pmol/mg protein/10 min, occurred in NA-induced cAMP accumulation, with no further increase after longer durations of ischemia. The mechanisms modulating the increase in cortical NA-inducible cAMP accumulation with a maximum response after 20 min of ischemia remain to be defined. PMID- 6296323 TI - Intrathecal synthesis of virus-specific oligoclonal IgG, IgA and IgM antibodies in a case of varicella-zoster meningoencephalitis. AB - Varicella-zoster (VZ) virus meningoencephalitis was diagnosed in a 72-year-old man without other clinical signs of VZ infection, on the basis of intrathecal virus-specific IgG, IgA and IgM antibody responses demonstrated by imprint immunofixation (IIF) and by serological analyses of serum and CSF. The intrathecally produced antibodies displayed oligoclonal characteristics. The intrathecal production of VZ-IgG and -IgA antibodies persisted throughout the observation period of 20 months, while that of VZ-IgM antibodies was not detectable later than 3 months after onset. Part of the intrathecally produced VZ IgG and -IgA antibody populations, but no IgM antibodies, were shown to cross react with herpes simplex virus. Oligoclonal IgG bands were demonstrated in the CSF throughout the observation period. The bulk of the IgG bands was shown to represent VZ-specific antibodies. PMID- 6296325 TI - The cerebellar medulloblastoma and its relationship to primitive neuroectodermal tumors. AB - A simple classification system for central nervous system neoplasms occurring primarily in infancy and childhood and largely composed of undifferentiated neuroepithelial cells is proposed. Classification is based upon appearance of the tumor as determined by light microscopy, immunocytochemical techniques, and ultrastructural features without consideration for site of origin. This classification is based on the concept that neoplastic transformation of primitive neuroepithelial cells in subependymal zones at all levels of the central nervous system or pineal body may develop into tumors largely composed of similar cells. It therefore seems appropriate to call these neoplasms primitive neuroectodermal tumors and to use descriptive terms to indicate the direction of cellular differentiation, when it has occurred. Proposed terminology for the five subtypes of undifferentiated neuroepithelial round cell tumors is as follows: 1) Primitive neuroectodermal tumor, not otherwise specified (PNET, NOS), 2) PNET with glial differentiation, 3) PNET with ependymal differentiation, 4) PNET with neuronal differentiation, and 5) PNET with multi- or bipotential differentiation. If the tumor is located in the cerebellum, medulloblastoma may be added in parentheses; if in the pineal body, pineal parenchymal neoplasm may be added. PMID- 6296324 TI - Time-dependence of, and effects of inhibition and cellular aging on, chloride efflux across erythrocyte membranes in Huntington's disease. AB - Previous studies have demonstrated an increased rate of chloride transport across erythrocyte membranes in Huntington's disease, a process regulated at the external side of the major transmembrane protein Band 3. A marked effect of time was noted in 3 Huntington's disease samples that were studied more than 4 h after obtaining the blood. In order to study anion transport more closely in Huntington's disease and the apparent time-dependence of chloride efflux in this disorder, we have performed several sets of experiments. Chloride transport in Huntington's disease erythrocytes was found to be extremely sensitive to time with the efflux rate constant decreasing by approximately 30% over a 24 h period. Chloride transport in control cells was unaffected by time. Inhibition studies with the specific anion transport blocker 4,4'-diisothiocyanto-2,2'-disulfonic acid stilbene (DIDS) demonstrated that the same degree of inhibition of chloride transport could be achieved at a much lower concentration of DIDS in Huntington's disease than in controls. Comparison of chloride efflux in fractions enriched in young and old erythrocytes, respectively, obtained by density centrifugation of fresh blood, demonstrated that only in the young fraction of cells was chloride efflux diminished with time in Huntington's disease. Chloride transport in in vivo aged Huntington's cells or both young and old control cells was essentially not dependent on time. These results are discussed in terms of proposed molecular mechanisms for neuronal loss in this disorder. The alterations in chloride efflux in extraneural erythrocytes are consistent with a proposed cell-surface membrane defect involving a protein in Huntington's disease. PMID- 6296326 TI - Responses of cortical neurons to stimulation of corpus callosum in vitro. AB - 1. An in vitro slice preparation of rat cingulate cortex was used to analyze the responses of layer V neurons to electrical stimulation of the corpus callosum (CC). In addition, synaptic termination of callosal afferents with layer V neurons was evaluated electron microscopically to provide a structural basis for interpreting some of the observed response sequences. 2. Layer V neurons had a resting membrane potential (RMP) of 60 +/- 0.68 (SE) mV, an input resistance of 47 +/- 4.74 M omega, a membrane time constant of 4.37 +/- 0.51 ms, an electrotonic length constant of 1.38 +/- 0.25, and produced spontaneous action potentials that were 50 +/- 0.3 mV in amplitude. Intracellular depolarizing current pulses evoked spikes that were sometimes associated with low-amplitude (2 5 mV) depolarizing (5-10 ms in duration) and hyperpolarizing (10-20 ms in duration) afterpotentials. 3. A single stimulus of increasing intensities to the CC produced one of the following response sequences: a) antidromic spike and an excitatory postsynaptic potential (EPSP), which initiated one or more spikes; b) antidromic spike, EPSP-evoked action potentials, and a hyperpolarization, which may have represented an intrinsic cell property or inhibitory synaptic activity; c) EPSP and evoked spikes only; d) high-amplitude EPSP with an all-or-none burst of action potentials. 4. Antidromically activated (AA) neurons always produced EPSPs in response to CC stimulation. When compared with nonantidromically activated neurons, AA cells had a more negative RMP, greater electrotonic length constant (LN), higher ratio of dendritic to somatic conductance (rho), and formed shorter duration, callosal-evoked EPSPs. 5. Neurons in anterior cingulate cortex produced EPSPs of longer duration than did those in posterior cortex (50 +/- 3.57 versus 26 +/- 1.56 ms, respectively). EPSPs in anterior neurons also had a higher maximum amplitude (20.5 +/- 1.0 versus 11.5 +/- 0.79 mV) and longer time to peak (11.6 +/- 2.2 versus 8.2 +/- 0.8 ms). 6. Electron microscopy of Golgi-impregnated neurons following contralateral lesions demonstrated that both pyramidal and nonpyramidal neurons received direct callosal afferents. Synaptic termination of callosal axons with the apical dendritic trees of anterior pyramidal cells was 6 times greater than it was with posterior pyramidal neurons. 7. EPSP shape differences in anterior and posterior neurons may be partially accounted for by the density and distribution of callosal afferents to these two cortices. PMID- 6296327 TI - Tonic and phasic synaptic input to spinal cord motoneurons during fictive locomotion in frog embryos. AB - 1. In curarized, late developmental stage Xenopus embryos, episodes of rhythmic motor root discharge, termed fictive swimming (17), may be evoked by touch or by dimming the lights, as in unparalyzed animals. Motoneurons are tonically depolarized throughout each episode, are phasically excited to fire 1 spike per cycle, and receive a midcycle inhibitory postsynaptic potential (IPSP) in phase with motor root activity on the opposite side. 2. Rostral hemisection of the spinal cord abolishes motor root discharge on the operated side caudal to the cut but leaves activity on the intact side unaffected. In motoneurons, the tonic depolarization is abolished on the hemisected side but is still present on the intact side. This is evidence that the tonic depolarization is a descending drive. 3. Midcycle IPSPs normally seen in motoneurons during fictive swimming are abolished by rostral hemisection of the opposite side of the cord but are still recorded on the cut side. The simplest conclusion is that the inhibitory interneurons responsible lie on the opposite side of the spinal cord to the motoneurons they inhibit, and so represent a reciprocal inhibitory pathway. 4. The phasic excitatory postsynaptic potentials (EPSPs), which drive motoneuron spikes during swimming, are still present on the intact side of a rostrally hemisected cord but are abolished on the operated side. We conclude that the excitatory interneurons responsible lie on the same side of the cord as the motoneurons they excite. PMID- 6296328 TI - Electrophysiological properties of neocortical neurons in vitro. AB - 1. Intracellular recordings were obtained from neurons of the guinea pig sensorimotor cortical slice maintained in vitro. Under control recording conditions input resistances, time constants, and spiking characteristics of slice neurons were well within the ranges reported by other investigators for neocortical neurons in situ. However, resting potentials (mean of -75 mV) and spike amplitudes (mean of 93.5 mV) were 10-25 mV greater than has been observed in intact preparations. 2. Current-voltage relationships obtained under current clamp revealed a spectrum of membrane-rectifying properties at potentials that were subthreshold for spike generation. Ionic and pharmacologic analyses suggest that subthreshold membrane behavior is dominated by voltage-sensitive, very slowly inactivating conductances to K+ and Na+. 3. Action potentials were predominantly Na+ dependent under normal conditions but when outward K+ currents were reduced pharmacologically, it was possible, in most cells, to evoke a non Na+-dependent, tetrodotoxin-(TTX) insensitive spike, which was followed by a prominent depolarizing after-potential. Both of these events were blocked by the Ca2+ current antagonists, Co2+ and Mn2+. 4. A small population of neurons generated intrinsic, all-or-none burst potentials when depolarized with current pulses or by synaptic activation. These cells were located at a narrow range of depths comprising layer IV and the more superficial parts of layer V. 5. Spontaneous excitatory synaptic potentials appeared in all neurons. Spontaneous inhibitory events were visible in only about 10% of the cells, and in those cases apparently reversed polarity at a level slightly positive to resting potential. Stimulation of the surface of the slice at low intensities evoked robust and usually concurrent excitatory and inhibitory synaptic potentials. Unitary inhibitory postsynaptic potentials (IPSPs) reversed at levels positive to rest. Stronger stimulation produced a labile, long-duration, hyperpolarizing IPSP with a reversal potential 15-20 mV negative to the resting level. 6. Neocortical neurons in vitro retain the basic membrane and synaptic properties ascribed to them in situ. However, the array of passive and active membrane behavior observed in the slice suggests that cortical neurons may be differentiated by specific functional properties as well as by their extensive morphological diversity. PMID- 6296329 TI - Mechanisms underlying pattern generation in lobster stomatogastric ganglion as determined by selective inactivation of identified neurons. III. Synaptic connections of electrically coupled pyloric neurons. AB - 1. The pyloric dilator (PD) and anterior burster (AB) neurons in the pyloric system of the lobster stomatogastric ganglion are electrically coupled and synchronously active. We have used the lucifer yellow photoinactivation technique to separate the connections made by the PD motor neurons from those made by the AB interneuron. 2. Photoinactivation of either the two PD neurons or the single AB neuron allowed us to separate the compound inhibitory postsynaptic potentials (IPSPs) in the lateral pyloric (LP) and pyloric (PY) motor neurons resulting from synchronous PD and AB activity into AB-evoked and PD-evoked components. These IPSPs have different time courses, reversal potentials, ion selectivities, and pharmacological properties. 3. Photoinactivation and membrane-potential manipulations indicated that a readily observable IPSP recorded in the AB neuron and correlated with action potentials in the LP neuron is actually an electrotonic potential due to an LP-evoked IPSP in the PD neurons. 4. Selective inactivation of either the two PD neurons or the AB neuron revealed that the IPSP recorded in the ventricular dilator (VD) motor neuron is due solely to AB released transmitter. 5. The electrical coupling potentials measurable between the AB, PD, and VD neuron somata are due to direct electrical coupling between all of these neurons. 6. Circuit analysis and transmitter identification may be complicated by electrical coupling. We suggest that the presence of electrical coupling between nonidentical neurons may provide a new mechanism that allows changes in synaptic characteristics among neurons within a "hard-wired" circuit. PMID- 6296330 TI - Protection of opiate receptors in NG108-15 against modification by N ethylmaleimide. AB - Two different -SH groups associated with the opiate receptors of the mouse neuroblastoma X rat glioma hybrid NG108-15 have been identified. Modification of these by N-ethylmaleimide (NEM) (presumed to be via alkylation) or by para chloromercuribenzoic acid (presumed to be via formation of mercury adducts) decreases the binding of both opiate agonists and antagonists to these receptors. Agonist binding is more sensitive than antagonist binding to modification by NEM. Losses in antagonist binding are accounted for totally by decreases in the number of binding sites; there are no corresponding losses in antagonist affinity. Losses of antagonist binding exhibit a pseudo-first order rate constant; the modification of only one such group completely destroys the binding site. Both agonists and antagonists protect against modification of this group by NEM. Sodium and lithium, but not GTP, also protect this group, indicating that the action of these monovalent cations is directly on the receptor moiety. Losses in agonist binding stem not only from decreases in receptor number but also from selective losses in affinity. This -SH group appears to be different from the one at the binding site as sodium, GTP, and antagonist ligands do not protect against losses in agonist affinity. Agonist high affinity also is lost in a pseudo-first order fashion indicating that an alteration of only one -SH group per receptor complex is sufficient to produce this effect. The possible roles of two sulfhydryls in opiate receptor function are discussed. PMID- 6296331 TI - Regional distribution of calcium- and cyclic adenosine 3':5'-monophosphate regulated protein phosphorylation systems in mammalian brain. I. Particulate systems. AB - The regional distribution of phosphoproteins whose phosphorylation is regulated either by cyclic AMP or by calcium in combination with calmodulin or phospholipid has been investigated in particulate preparations from rat CNS. About 30 distinct phosphoproteins were observed. These phosphoproteins exhibited widely different patterns of regional distribution. Based upon distribution patterns, we have divided these phosphoproteins into three categories: category A, phosphoproteins found in all parts of the CNS in approximately equal amounts; category B, phosphoproteins which are widely distributed within the CNS but show large regional variations; and category C, phosphoproteins which show a highly restricted regional distribution. We have tentatively interpreted the results on particulate phosphoproteins in the following way: some are present in all or nearly all brain cells, others are present only in certain classes of brain cells, and still others have an even more limited distribution, being present in only a single type of brain cell. The regional distribution of particulate protein kinase activity was also examined. Calcium/calmodulin-dependent protein kinase activity had a marked regional distribution, whereas cyclic AMP-dependent protein kinase activity was more evenly distributed. Calcium/phospholipid dependent protein kinase activity was barely detectable under the experimental conditions used. This investigation thus demonstrates striking differences in the regional distribution of particulate protein phosphorylation systems in mammalian brain. These regional differences may reflect highly specific functional roles for certain of these protein phosphorylation systems. Similar conclusions concerning cytosolic protein phosphorylation systems are described in the accompanying paper. PMID- 6296332 TI - Regional distribution of calcium- and cyclic adenosine 3':5'-monophosphate regulated protein phosphorylation systems in mammalian brain. II. Soluble systems. AB - The regional distribution of phosphoproteins whose phosphorylation is regulated either by cyclic AMP or by calcium in combination with calmodulin or phospholipid has been investigated in soluble preparations from rat CNS. About 40 distinct phosphoproteins were observed. These cytosolic phosphoproteins exhibited widely different patterns of regional distribution. Based upon distribution patterns, we have divided these phosphoproteins into three categories: category A, phosphoproteins found in all parts of the CNS in approximately equal amounts; category B, phosphoproteins which are widely distributed within the CNS, but which show large regional variations; and category C, phosphoproteins which show a highly restricted regional distribution. We have tentatively interpreted the results on cytosolic phosphoproteins in the following way: some are present in all or nearly all brain cells, others are present only in certain classes of brain cells, and still others have an even more limited distribution, being present in only a single type of brain cell. The regional distribution of soluble protein kinase activity was also studied. Calcium/phospholipid-dependent protein kinase and calcium/calmodulin-dependent protein kinase had marked regional distributions. Cyclic AMP-dependent protein kinase was more evenly distributed throughout the CNS. This investigation thus demonstrates striking differences in the regional distribution of cytosolic protein phosphorylation systems in mammalian brain. These regional differences may reflect highly specific functional roles for certain of these protein phosphorylation systems. Similar conclusions concerning particulate protein phosphorylation systems are described in the preceding paper. PMID- 6296333 TI - Background and stimulus-induced patterns of high metabolic activity in the visual cortex (area 17) of the squirrel and macaque monkey. AB - We have used 2-deoxy-D-[14C]glucose (2-DG) autoradiography and cytochrome oxidase histochemistry to examine background and stimulus-induced patterns of metabolic activity in monkey striate cortex. In squirrel monkeys (Saimiri sciureus) that binocularly or monocularly viewed diffuse white light or binocularly viewed bars of many orientations and spatial frequencies, 2-DG consumption was not uniform across the cortex but consisted of regularly spaced radial zones of high uptake. The zones extended through all laminae except IVc beta and, when viewed tangentially, formed separate patches 500 microns apart. The cytochrome oxidase stain in these animals also revealed patches of high metabolism which coincided with the 2-DG patches. Squirrel monkeys binocularly viewing vertical stripes showed parallel bands of increased 2-DG uptake in the cortex, while the cytochrome label in these animals remained patchy. When monkeys were kept in the dark during 2-DG exposure, 2-DG-labeled patches were not seen but cytochrome oxidase-positive patches remained. In macaque (Macaca nemestrina) monkeys, binocular stimulation with many orientations and spatial frequencies produced radial zones of high 2-DG uptake in layers I to IVa and VI. When viewed tangentially, these zones formed a dots-in-rows pattern with a spacing of 350 X 500 microns; cytochrome oxidase staining produced an identical pattern. Macaca differed from Saimiri in that monocular stimulation labeled alternate rows. These results indicate that there are radial zones of high background metabolism across squirrel and macaque monkey striate cortex. In Saimiri these zones do not appear to be related to an eye dominance system, while in Macaca they do. The presence of these zones of high metabolism may complicate the interpretation of 2-DG autoradiographs that result from specific visual stimuli. PMID- 6296334 TI - Photoaffinity labeling of benzodiazepine receptors causes altered agonist antagonist interactions. AB - Previous studies have shown that [3H]flunitrazepam forms irreversible cross-links with brain tissue when exposed to ultraviolet irradiation. Comparison of the amount of [3H]flunitrazepam irreversibly incorporated and the number of benzodiazepine binding sites blocked after photolabeling has indicated that several binding sites are inactivated for each molecule of [3H]flunitrazepam incorporated. To learn the cause of this discrepancy, binding to the benzodiazepine binding sites has been examined using several radiolabeled benzodiazepine antagonists. Binding of a beta-carboline ester, CGS-8216, and Ro 15-1788 was not altered by photolabeling; however, displacement studies revealed that photolabeling converted a homogeneous set of benzodiazepine binding sites into two subsets: one of high affinity (unaltered sites) and one of low affinity. The low affinity sites could be detected by displacement studies of antagonist binding by benzodiazepines, and conversion to a low affinity form accounts for the discrepancy observed after photolabeling using [3H]flunitrazepam as ligand. PMID- 6296335 TI - Faster assessment of patients receiving unnecessary thyroid treatment: concise communication. AB - Forty-five consecutive patients on thyroid hormone treatment without obvious indication were evaluated. Twenty-five of these cases were found to have no evidence of thyroid disease. Biochemical testing was not helpful in making the diagnosis of hypothyroidism in the majority of thyroid-treated hypothyroid patients. Normal technetium images were obtained in 25 patients, 22 of which had no thyroid disease. In contrast, abnormal technetium images were obtained in 20 patients, 16 of whom were thought to be hypothyroid, and one of whom developed a goiter within 2 mo after discontinuing levothyroxine. The use of technetium imaging seems useful for the rapid (20 min) evaluation of those patients likely to benefit from discontinuing thyroid medication. PMID- 6296336 TI - Gold-195m, an ultra-short-lived generator-produced radionuclide: clinical application in sequential first pass ventriculography. AB - Gold 195m (Au-195m) has a half-life of 30.5 sec and can be produced at the bedside from the parent mercury-195m (T 1/2 = 41.6 hr). The generator produced sterile pyrogen-free Au-195m with mercury breakthrough of 0.75 +/- 0.09 (s.e.m.) muCi per mCi of Au-195m. Approximately 20 to 25 mCi of Au-195m was produced per elution from a generator containing 155 mCi of Hg-195m. We compared first-pass resting Tc-99m angiograms with Au-195m angiograms in 28 patients. The correlation coefficient between the two studies was 0.92 over an ejection-fraction range from 0.22 to 0.83. In addition, we tested the reproducibility of Au-195m first-pass angiograms by performing two studies 3 min apart. In 25 patients with ejection fractions ranging from 0.20 to 0.78, the correlation coefficient between such pairs was 0.93. The nuclide is reliably and reproducibly produced, and its short half-life allows the performance of background-free sequential first-transit studies with unusually low radiation exposure to the patient. PMID- 6296337 TI - Determination of left-ventricular volume from first-pass kinetics of labeled red cells. AB - A mathematical model is presented for the dynamics of a bolus of technetium-99m labeled red blood cells through the left ventricle. It is used to correct for attenuation the count rate observed over the left ventricle during a conventional gated blood-pool study. The left-ventricular volumes are calculated from the corrected count rates and expressed as a percentage of total blood volume, or in absolute terms if a blood sample is obtained. The procedure is applied to a number of patients with nonvalvular cardiac disease. Cardiac volumes determined by the method are found to correlate well (r = 0.98) with those determined by contrast left ventriculography. The method is simple, requires no special equipment, and can be applied with existing computer software. PMID- 6296339 TI - Effects of dietary fiber on fecal mucinase and beta-glucuronidase activity in rats. AB - Mucinase and beta-glucuronidase enable colon bacteria to degrade protective mucins and recycle glucuronide conjugates of toxins and carcinogens. The response of these bacterial enzymes to dietary fiber was studied in the laboratory rat. Fiber-free basal diet was mixed with guar gum, pectin, carrageenan, or cellulose at levels of 5 and 15%. These diets were fed for 21 days to groups of six male Fischer-344 rats having an average weight of 150 g. Mucinase and beta glucuronidase activities were assayed in fresh rat feces. Rats fed 15% guar gum or pectin gained significantly (P less than 0.05) less weight than the other rats. Mucinase specific activity was highest in the fiber-free diet group and lowest in the 15% guar gum group. Total daily output of mucinase was highest in rats fed fiber-free diet or cellulose and lower in rats fed more readily fermentable fiber. Specific activity and total output of beta-glucuronidase were highest in rats fed fiber-free diet and significantly lower in those fed 15% fiber diets. These data are consistent with the hypothesis that some kinds of dietary fiber may play a role in the etiology of intestinal disease. PMID- 6296340 TI - Bioavailability to rats of selenium in various tuna and wheat products. AB - Bioavailability of selenium (Se) in tuna and wheat at various stages of processing was studied in rats. The protein source of the rat diets was torula yeast with Se supplied by either raw, precooked or canned tuna, or whole wheat flour, whole wheat bread or bran. Sodium selenite was used as the standard. Each Se source was fed at three levels: 0.05, 0.10 and 0.15 ppm. By using increase in glutathione peroxidase (GSH-Px) activity in liver, kidney and whole blood as an indicator of bioavailability, no differences were found among the three tuna products or among the three wheat products tested. However, significantly lower GSH-Px activity was found in the combined tuna groups as compared to the combined wheat groups, suggesting that selenium in wheat was more available than that in tuna. There was a significant increase in the liver Se content of rats fed all levels of Se in canned tuna and in kidney, blood and muscle Se of rats fed 0.10 and 0.15 ppm Se in canned tuna in comparison to the tissue Se content in rats fed these same levels of Se in raw or precooked tuna. Since this did not correspond with an increase in GSH-Px activity it was concluded that it did not represent increased bioavailability of canned tuna. Thus, food processing does not appear to affect Se availability, but Se appears to be more available in wheat than tuna. PMID- 6296338 TI - Arginine utilization of young rats fed diets with simple versus complex carbohydrates. AB - We proposed that increasing dietary complex carbohydrates would, by increasing fecal nitrogen loss and thus decreasing urinary nitrogen and the need for urea synthesis, ameliorate the effects of arginine deficiency. Two experiments using male, weanling, Sprague-Dawley rats with ad libitum access to water and isocaloric, isonitrogenous, 19% L-amino acid diets were carried out to test this hypothesis. The first was performed in two identical blocks of a 4 X 4 factorial experiment each with six rats per group. Four levels of arginine were fed: 0.0, 0.4, 0.8 and 1.2% of diet with four variations in dietary carbohydrates: sucrose alone, replacement of 2/3 of sucrose with dextrin, and 5 g of wheat bran or guar gum added to 100 g of the sucrose-based diets. Varying dietary carbohydrate quality did not influence growth of rats fed the 1.2% arginine diets, however, when added to the diet with 0.0 arginine, guar gum but not dextrin or wheat bran increased growth. This guar gum-induced weight increase was associated with partial reversal of arginine-deficiency-induced liver lipidosis and orotic aciduria and with higher fecal nitrogen and lower urinary nitrogen. The specificity of this guar gum effect was tested in a second experiment in which guar gum was added to diets containing 33% of the control level of arginine, histidine, leucine, methionine or phenylalanine. Only rats fed the diet deficient in arginine showed significant improvement in growth with added guar gum. We conclude that dietary guar gum but not dextrin or wheat bran, by increasing fecal nitrogen loss and decreasing the need for urea synthesis, partially ameliorates the effects of a low arginine intake. The practical application of feeding guar gum to alleviate certain ammonia-toxicity-related medical conditions is discussed. PMID- 6296341 TI - Effect of feeding alfalfa and refined plant fibers on the toxicity and metabolism of T-2 toxin in rats. AB - Experiments were conducted to determine the effect of dietary fibers on T-2 toxicosis in rats. Weanling rats were fed varying levels of cellulose, hemicellulose, lignin and pectin with and without T-2 toxin (3 micrograms/g feed) for 2 weeks. Only lignin showed promise of overcoming feed refusal and growth depression in animals fed T-2 toxin. Further experiments feeding alfalfa meal (0, 5, 10, 15, 20 or 25%) with and without T-2 toxin indicated that this lignin-rich feedstuff could largely overcome feed refusal and growth depression caused by the toxin. There was no effect of diet, however, on the activity of hepatic esterase, the enzyme believed to catabolize T-2 toxin. Rats were fed diets containing 0, 5, 12.5 or 20% alfalfa for 2 weeks and then dosed orally with [3H]T-2 toxin. Dietary alfalfa increased fecal excretion of 3H, whereas urinary excretion was unaffected. Residual 3H in kidney and muscle was reduced with alfalfa feeding when [3H]T-2 toxin was administered orally. Residual 3H in the digesta in the intestinal lumen increased. Alfalfa feeding was found to reduce intestinal transit time. It was concluded that the feeding of alfalfa reduced T-2 toxicosis in rats by binding the toxin in the intestinal lumen thereby promoting fecal excretion. PMID- 6296342 TI - Folate deficiency in rats fed amino acid diets. AB - Growth rate, hematological changes, serum, erythrocyte and liver folate levels, and urinary excretion of formiminoglutamic acid (FIGLU) were measured in rats fed p-aminobenzoic acid (PABA)-free, sulfonamide-supplemented purified amino acid diets with and without folic acid and fiber, individually and in combination. Fiber had no effect on growth but was necessary to prevent mortality in the absence of folic acid. Folic acid did not affect growth during the first 40 days, but after this period rats failed to gain weight in the absence of folic acid. Although blood hematocrit was lower when the diet was devoid of fiber and folic acid and leucocyte counts were lower when the diets were devoid of folic acid, the hematological values remained within normal physiological limits for healthy rats of comparable age. FIGLU excretion was increased and serum, erythrocyte and liver folate levels were depressed when rats were fed folic acid-free diets for 28 days. Neutrophil hypersegmentation was clearly evident after 28 days of feeding the folic acid-free diets. The use of an amino acid diet without PABA but containing 5% fiber and 1% succinylsulfathiazole is a useful system to study chronic folic acid deficiency in the rat. PMID- 6296343 TI - Respiratory morbidity among workers in an amosite asbestos insulation plant. PMID- 6296344 TI - Functional differences between ethacrynic acid and ouabain on glucose secretion in rat submandibular gland. PMID- 6296345 TI - X-ray microanalysis of the Michaelis-Gutmann bodies of malakoplakia. AB - The Michaelis-Gutmann (MG) inclusion bodies of three cases of malakoplakia (prostate, testis, colon) were studied by X-ray microanalysis to determine their elemental composition. Calcium and phosphorus were consistently found. Iron was detected in a few bodies. No other elements were detected. The electrondense laminations were of similar composition to the core material. Small aggregates of electron-dense material containing calcium and phosphorus were also occasionally seen in phagolysosomes. These observations are consistent with view that MG bodies arise by a process of phagolysosomal coalescence and mineralisation. PMID- 6296346 TI - Biochemical and immunohistochemical studies on collagenase in resorbing bone in tissue culture. A novel hypothesis for the mechanism of bone resorption. PMID- 6296347 TI - Phase contrast microscopic evaluation of subgingival plaque in combination with either conventional or antimicrobial home treatment of patients with periodontal inflammation. PMID- 6296348 TI - Topical chemical antimicrobial therapy in the management of the subgingival microflora and periodontal disease. PMID- 6296349 TI - Human clinical and histologic responses to Durapatite implants in intraosseous lesions. Case reports. AB - The healing response following implantation of a nonresorbable ceramic (durapatite) into human periodontal osseous defects was evaluated clinically and histologically. Four tooth-containing blocks were obtained from four patients who had received durapatite implants in osseous defects, each exceeding 4 mm in depth. Each patient was seen for 5 to 13 postsurgical maintenance visits. Teeth in block section were removed between 8 weeks and 8 months postgraft surgery. Clinical evaluation of the repair process demonstrated that pocket depth decreased in all four cases. Histological evaluation of the repair process showed no indication of new periodontal attachment, osteogenesis or cementogenesis, in the host tissues adjacent to the graft particles. Pocket closure appeared to occur by means of a long junctional epithelium and connective tissue adhesions. There was minimal or no evidence of inflammation in all sections associated with the implant. The graft material therefore acted as a biocompatible foreign body within the gingival tissue. PMID- 6296350 TI - The ratio of cardiac to vascular beta-receptor blockade of atenolol and propranolol in spontaneously hypertensive rats in vivo. AB - Ratios of cardiac to vascular beta-receptor blockade (C:V ratios) of atenolol and propranolol were determined in pentobarbital anesthetized spontaneously hypertensive (SHR) rats. These values and the cardiovascular responses to isoproterenol were compared with those of age-matched normotensive control (Donryu, DON) rats reported previously. Mean blood pressure (BP) and heart rate (HR) were recorded. Full dose-response curves for isoproterenol were constructed in BP and HR. In vivo pD2 values of isoproterenol (potencies), were not different between DON and SHR rats, although the maximal responses to isoproterenol (efficacy) were significantly suppressed in SHR rats. C:V ratios of atenolol and propranolol in SHR rats were 7.95 and 0.44, respectively. As the values in DON rats were 9.77 and 1.20, respectively, C:V ratios of atenolol and propranolol were decreased in SHR rats. Markedly decreased C:V ratios of propranolol in SHR rats may explain its relatively weak antihypertensive action. PMID- 6296351 TI - [Monoaminergic synapses in the brain--regulation and ontogenetic development]. PMID- 6296352 TI - [A simple and rapid method of radioimmunoassay of cyclic AMP with the cell harvester]. PMID- 6296353 TI - Lysine and polylysine: correlation of their effects on polyphosphoinositides in vitro with ototoxic action in vivo. AB - Low concentrations of poly-L-lysine, a polycationic hydrophilic molecule, caused a large expansion of polyphosphoinositide monolayers and produced a significant loss of cochlear microphonic potentials in perilymphatic perfusions in the guinea pig. In contrast, the monomeric L-lysine had only slight effects on polyphosphoinositide monolayers and did not affect cochlear microphonic potentials even at concentrations as high as 10 mM. These data substantiate the hypothesis that the expansion of polyphosphoinositide monolayers by a drug is an indicator of its ototoxicity. PMID- 6296354 TI - Alcohol modulation of drug binding to the channel sites of the nicotinic acetylcholine receptor. PMID- 6296355 TI - Characterization of the effect of quinidine on Na transport by the toad and turtle bladders. AB - Quinidine inhibits Na transport by the toad and turtle bladder. This effect of quinidine is thought to be mediated by an increase in cytosolic calcium. In the present study, we characterized the effect of quinidine on Na transport by the toad and turtle bladders. Quinidine induced a release of calcium by turtle liver mitochondria. Quinidine inhibited Na transport by increasing the resistance of the active pathway to Na transport without affecting the electromotive force. Amphotericin B addition to the mucosal solution partially reversed the inhibitory effect of quinidine on Na transport, thus suggesting that quinidine decreases Na transport by decreasing the permeability of luminal membrane to Na. The effect of amiloride was unaltered in the presence of quinidine. Vasopressin failed to stimulate Na transport in the presence of quinidine, suggesting that the drug interferes with the natriferetic effect in addition to interfering with the hydrosmotic effect. The effect of quinidine was not prevented by inhibition of cyclooxygenase system or mitochondrial inhibition, thus suggesting that alterations in prostaglandin release or mitochondrial function are not involved in the inhibition of Na transport by quinidine. PMID- 6296356 TI - I. Pharmacological studies with derivatives of 2-aminotetralin, benzhydro[f]quinoline and clonidine suggest a pharmacological identity between peripheral and central alpha-2 adrenoceptors. AB - A series of hydroxy 2-aminotetralins, benzhydro[f]quinolines and clonidine were used to determine whether a pharmacological similarity could be demonstrated between presynaptic alpha adrenoceptors which modulate autonomic transmission in the guinea pig. Compounds were assayed on isolated field stimulated guinea-pig ilea (GPI) to determine their inhibitory activities on cholinergic transmission. Inhibition of noradrenergic transmission was determined by assaying compounds on isolated field stimulated guinea-pig atria. 2-Aminotetralins, benzhydro[f]quinolines and clonidine impaired cholinergic and noradrenergic transmission by interacting with presynaptic alpha-2 adrenoceptors. A correlation (r = 0.95; P less than .05) was demonstrated between the activity of a compound on the GPI and guinea-pig atria. IC50 values obtained on GPI were correlated with previously reported IC50 values obtained for binding [3H]clonidine sites in homogenates of calf frontal cortex. A significant correlation was demonstrated (r = 0.99; P less than .05). These data suggest that alpha-2 adrenoceptors localized on guinea-pig atria and GPI are pharmacologically similar. In addition, a similar structure activity relationship was demonstrated for presynaptic alpha-2 adrenoceptors on GPI and binding sites labeled by [3H]clonidine in the calf frontal cortex. PMID- 6296357 TI - II. Pharmacological studies with derivatives of 2-aminotetralin, benzhydro[f]quinoline, benzhydro[g]quinoline, apomorphine and clonidine suggest a pharmacological dissimilarity between peripheral presynaptic dopamine receptors and alpha-2 adrenoceptors. AB - This study demonstrates that presynaptic dopamine receptors and alpha-2 adrenoceptors are pharmacologically different. A series of 2-aminotetralins, benzhydro[f]quinolines, benzhydro[g]quinolines, apomorphine and clonidine were studied to determine if they could stimulate presynaptic alpha-2 adrenoceptor and dopamine receptors. Presynaptic dopamine receptor activity was observed in di- and monohydroxy derivatives of 2-aminotetralins, dihydroxy derivatives of benzohydro[f]quinolines and benzohydro[g]quinolines and apomorphine. The greatest presynaptic dopamine receptor activity was observed with agents which maintained the dopamine moiety in the trans coplanar conformation. In contrast to these observations 1) monohydroxy derivatives of 2-aminotetralines were devoid of presynaptic alpha-2 adrenoceptor activity and 2) both cis and trans isomers of dihydroxy derivatives of benzohydro[f]quinolines and benzohydro[g]quinolines exhibited significant presynaptic alpha-2 adrenoceptors activity. These data suggest that presynaptic alpha-2 adrenoceptors and dopamine receptors represent separate functional entities. A discussion on the structure activity relationship associated with presynaptic alpha-2 adrenoceptor and dopamine receptor is provided. PMID- 6296358 TI - Relative involvement of receptor subtypes in opioid-induced inhibition of gastrointestinal transit in mice. PMID- 6296359 TI - Identification and localization of opioid receptors in the opossum lower esophageal sphincter. PMID- 6296360 TI - Marijuana: interaction with the estrogen receptor. AB - Crude marijuana extract competed with estradiol for binding to the estrogen receptor of rat uterine cytosol. Condensed marijuana smoke also competed with estradiol for its receptor. Pure delta 9-tetrahydrocannabinol, however, did not interact with the estrogen receptor. Ten delta 9-tetrahydrocannabinol metabolites also failed to compete with estradiol for its receptor. Of several other common cannabinoids tested, only cannabidiol showed any estrogen receptor binding. This was evident only at very high concentrations of cannabidiol. Apigenin, the aglycone of a flavinoid phytoestrogen found in cannabis, displayed high affinity for the estrogen receptor. To assess the biological significance of these receptor data, estrogen activity was measured in vivo with the uterine growth bioassay, using immature rats. Cannabis extract in large doses exhibited neither estrogenic nor antiestrogenic effects. Thus, although estrogen receptor binding activity was observed in crude marijuana extract, marijuana smoke condensate and several known components of cannabis, direct estrogenic activity of cannabis extract could not be demonstrated in vivo. PMID- 6296361 TI - Activation of central nervous system cholecystokinin receptors stimulates respiration in the cat. AB - Cholecystokinin-octapeptide (CCK-OP) was injected into the lateral brain ventricle of chloralose-anesthetized cats while monitoring pulmonary ventilation, mean blood pressure (BP) and heart rate (HR). Initial values for respiratory minute volume, BP and HR were 436 +/- 32 ml/min, 126 +/- 8 mm Hg and 204 +/- 9 beats/min, respectively. CCK-OP in doses of 8.8 x 10(-13) to 8.8 x 10(-10) M (i.e., 1 to 1000 ng) increased respiratory minute volume by +56 +/- 18 ml/min to +370 +/- 73 ml/min, respectively. This response was due to an increase in tidal volume (P less than .05) with no effect on respiratory rate. Most of these doses increased BP and HR (P less than .05). These responses were mimicked when CCK-OP was administered into and restricted to the lateral and 3rd ventricles, whereas no significant responses occurred with administration into the 4th ventricle. The highest dose of CCK-OP, when given i.v., produced no significant effects. Central nervous system injections (lateral ventricle) of CCK-OP in deafferented animals also produced respiratory and cardiovascular stimulation, but higher doses were required. Central nervous system injections of comparable doses of desulfated CCK OP or pentagastrin had no effect on respiratory minute volume, BP or HR. Proglumide (0.8 and 8.0 mg), a specific CCK receptor antagonist, prevented both respiratory and cardiovascular effects induced by CCK-OP. These doses of proglumide exerted no antagonism of CO2-induced respiratory stimulation. These data suggest that a specific receptor reacting with CCK-OP may be important for the control of respiration. PMID- 6296362 TI - Alterations of the [59Fe]ferric citrate biodistribution in hyperferremic mice after the administration of pyrophosphate and desferrioxamine. AB - One of the most efficient anions in enhancing the ability of desferrioxamine (DFO) to remove iron from transferrin in vitro has been shown to be pyrophosphate (PYP). To evaluate the in vivo effect of PYP in hyperferremic mice, the biodistribution of [59Fe]ferric citrate was studied after the i.p. administration of: 1) only saline in the control animals; 2) an aqueous solution of tetrasodium diphosphate (PYP; 40 gm/2 g of b.wt.); 3) desferral (DFO; 12 mg/20 g of b.wt.); and 4) PYP + DFO at the respective dosages shown above. The radioactivity in each organ, blood, urine and feces was measured and referred to as percentage of the injected dose. PYP administered alone acted as a weaker chelator of iron than DFO. The combined administration of DFO and PYP contributed more than DFO or PYP separately, to the increase of urinary excretion of 59Fe and to the significant decrease of the radioiron concentration in liver (.01 less than P less than .05). The above induced changes are not, however, the additive result of the separate effect of DFO and PYP. That observation would suggest that DFO + PYP combined in a unique treatment, interact with iron through a common reaction pathway and that PYP plays in vivo a synergistic role in that interaction. The kind of iron with which DFO + PYP interacts is then suggested to be the transferrin-bound iron located in extracellular spaces of tissues. PMID- 6296363 TI - Mechanism of morphine in increasing plasma cyclic GMP level in male mice. AB - Morphine given systemically or centrally increases the plasma concentration of cyclic GMP in male, ddY strain mice. Normorphine also increased the plasma cyclic GMP level, whereas the same dose of ketocyclazocine and SKF 10,047 had no effect. The effect of morphine on plasma cyclic GMP was mimicked by opioid peptides such as (D-Ala2, Met)-enkephalinamide, FK 33,824 or beta-endorphin. The effect of morphine and opioid peptides on plasma cyclic GMP was antagonized by naloxone, indicating the involvement of the opiate receptor. The increase in plasma cyclic GMP elicited by morphine was abolished by vagotomy and pretreatment with hexamethonium and atropine and was partly inhibited by pretreatment with phentolamine. Adrenalectomy and pretreatment with propranolol, which inhibited the increase in plasma cyclic AMP level elicited by morphine (Muraki et al., 1979), did not alter the cyclic GMP response to morphine. The development of tolerance to the cyclic GMP increase was observed in morphine-tolerant/dependent mice. These results suggest that morphine increases the plasma cyclic GMP level by activating the preganglionic parasympathetic tone via the stimulation of the opiate receptors, thereby increasing the generation of cyclic GMP through the muscarinic receptors on the effector cells. PMID- 6296364 TI - Effects of the sympathoadrenal system on vagally induced gastric acid secretion and mucosal blood flow in rats. AB - Effects of the sympathoadrenal system on vagally induced increases in gastric acid secretion and mucosal blood flow (MBF) were studied in anesthetized, gastric fistula rats. Greater splanchnic nerve stimulation reduced significantly both the gastric acid secretion and MBF. Stimulation of the splanchnic postganglionic nerve reduced to a greater extent the acid secretion than the MBF, while stimulation of the adrenal branch reduced both the acid secretion and MBF. Infusion of epinephrine also reduced both the acid secretion and MBF, but the reduction of MBF was the more prominent. Phentolamine, but not propranolol and alprenolol, reduced the inhibitory effects on both gastric functions of the splanchnic postganglionic nerve stimulation and adrenal branch stimulation or epinephrine infusion. These results suggest that catecholamines released from the splanchnic postganglionic nerve terminals inhibit gastric acid secretion through alpha adrenergic receptors in the stomach, independent of MBF, and that circulating catecholamines released from the adrenal medulla inhibit both the acid secretion and MBF through alpha adrenergic receptors in the stomach. The inhibition by greater splanchnic nerve stimulation is probably the sum of the effects induced by catecholamines released from the splanchnic postganglionic nerve terminals and from the adrenal medulla. PMID- 6296365 TI - Calcium antagonism of an opiate drug effect on an excitable cell membrane. AB - When frog sartorius muscles are exposed to methadone (2-4 X 10(-4) M) the action potential recorded intracellularly is depressed and eliminated in 3 to 4 hr. This is due to a decrease in the sodium conductance as measured by the maximum rate of rise of the action potential. When the calcium concentration in frog Ringer's solution (normally 1.08 mM) was either lowered to 0.54 mM or raised to 2 mM, the effect of methadone on excitability was unchanged. However, increasing the extracellular calcium to 4 mM decreased the action of methadone. As this would also increase the intracellular calcium concentration, in other experiments the free intracellular Ca+ concentration was raised by adding 0.4 mM caffeine to the Ringer's solution. This also antagonized the depressant action of methadone. The antagonistic action of caffeine was not due to a direct effect of caffeine on sodium conductance because the administration of caffeine by itself caused a 10% depression of the action potential maximum rate of rise and caffeine did not antagonize the action of tetrodotoxin, which is a specific sodium channel blocking agent. Finally, it was shown that the calcium ionophore, A23187, in low concentrations antagonized the depressant actions of methadone and meperidine on action potential production. It was concluded that increasing the intracellular free calcium concentration antagonized the depressant effect on excitability produced by opiate drugs acting on an intracellularly oriented opiate drug receptor. PMID- 6296366 TI - Properties of single calcium-activated potassium channels in cultured rat muscle. AB - 1. Properties of the Ca-activated K channel were studied in excised patches of surface membrane from cultured rat muscle cells using single channel recording techniques.2. Increasing the concentration of calcium at the intracellular membrane surface [Ca](i), increased both the frequency and effective duration of channel openings. Ca at the extracellular membrane surface was not sufficient to activate the channels.3. An approximate third power relationship (slope = 2.7) was observed between [Ca](i) and the percentage of time the channels spent in the open state.4. Both the frequency and effective duration of channel openings increased as the intracellular membrane surface was made more positive; the percentage of time spent in the open state increased e-fold for a 15 mV depolarization for low levels of activity.5. The percentage of time spent with 1, 2,...n channels open in membrane patches with n channels was described by the binomial distribution, suggesting that the channels opened and shut independently of one another.6. Single channel conductance (144 mM-K on both sides of the membrane) was essentially independent of membrane potential (-50 to +50 mV) and [Ca](i) (0.1 muM -1 mM), but did increase with temperature, from 100 pS at 1 degrees C to 300 pS at 37 degrees C.7. Channel activity occurred in apparent bursts, with the duration of the apparent bursts increasing with increasing [Ca](i).8. Two exponentials were required to describe the distribution of observed channel open times, suggesting two different open channel states of apparently normal conductance. The observed mean channel open time of these states at +30 mV was 0.34 and 2.2 msec with 0.1 muM-Ca(i) and was 0.47 and 6.9 msec with 0.5 muM-Ca(i).9. The channel occasionally entered an apparent third open channel state with a single channel current amplitude about 40% the amplitude of the normally observed single channel currents. The reduced conductance state was immediately preceded and followed by a normal conducting state.10. While the kinetics of the Ca-activated K channel appear complex, its large conductance and high Ca and voltage sensitivity suggest that it is uniquely suited to resist depolarizations of the cell membrane potential that are accompanied by increases in intracellular Ca. PMID- 6296367 TI - Studies of calcium channels in rat clonal pituitary cells with patch electrode voltage clamp. AB - 1. The properties of the Ca channel in tissue cultured clonal cells (GH(3)) isolated from a rat anterior pituitary tumour were studied with the patch electrode voltage-clamp technique.2. To isolate the current through the Ca channel, the currents through the Na channel, the delayed K channel and the Ca(2+) induced K channel were minimized by replacing the external Na(+) with TEA(+) and adding EGTA to the K-free solution inside the patch electrode.3. The selectivity ratios through the Ca channel with different cations were 2.7 (Ba(2+)):1.6 (Sr(2+)):1.0 (Ca(2+)) and the m(2) form of the activation kinetics and the relationships between the time constant and the membrane potential were common to the three divalent cations.4. The amplitude of the Ba(2+) current increased linearly with [Ba(2+)](o) up to 25 mM and thereafter tended to show saturation.5. The current-voltage relation showed a positive shift along the voltage axis as [Ba(2+)](o) increased, probably due to the screening effect of Ba(2+) on the negative surface charges.6. The time constant of activation as a function of the membrane potential showed a parallel shift as [Ba(2+)](o) was increased, suggesting that the activation kinetics were independent of the permeant ion concentration.7. The time constant of the tail current was consistent with m(2) kinetics for opening and closing of the Ca channel.8. The extrapolated ;instantaneous' tail current rapidly increased as the activating membrane potential became more positive and reached an apparent saturation at membrane potentials substantially more positive than the potential that gave the maximum peak inward current, and suggested that the single channel has a sigmoidal current-voltage relationship.9. The power density spectrum obtained during the steady-state inward Ba(2+) current had a cut-off frequency which was nearly voltage independent; this is expected if the fluctuation of the current originates from m(2) activation kinetics.10. The results of noise analysis suggest that the amplitude of the single Ca channel current was about 0.2 pA at 25 mM-Ba(2+) and 0.7 pA at 100 mM-Ba(2+) for membrane potentials in the vicinity of the maximum inward current. PMID- 6296369 TI - Effects of beta-adrenergic stimulation on calcium movements in rabbit aortic smooth muscle: relationship with cyclic AMP. AB - 1. The effects of isoprenaline (10(-6) M) on relaxation, unidirectional as well as net Ca(2+) fluxes, and cyclic AMP levels were investigated in rabbit aorta under the condition of high-K(+) depolarization in the presence of phentolamine (10(-5) M).2. Isoprenaline (10(-6) M) caused significant inhibition of Ca(2+) influx stimulated by 145 mM-K(+) (0 Na(+)) solution. The time courses of Ca(2+) influx inhibition and relaxation by isoprenaline were parallel. Isoprenaline also caused a significant inhibition of high-K(+)-induced gain in net Ca(2+) content.3. Ro 20-1724 (1 mM), a phosphodiesterase inhibitor, also caused relaxation and Ca(2+) influx inhibition in high-K(+)-depolarized rabbit aorta. Pre-treatment with Ro 20-1724 potentiated isoprenaline-induced Ca(2+) influx inhibition and relaxation.4. Isoprenaline and Ro 20-1724 each alone increased cyclic AMP levels. Furthermore pre-treatment with Ro 20-1724 caused potentiation of isoprenaline-induced increases in cyclic AMP levels.5. At submaximal concentration, D600 (10(-7) M) caused partial inhibition of high-K(+)-stimulated Ca(2+) influx and produced relaxation. However, unlike Ro 20-1724, it did not potentiate isoprenaline-induced Ca(2+) influx inhibition and relaxation. D600 does not increase cyclic AMP levels in smooth muscle.6. Dibutyryl cyclic AMP (1 mM), a lipid-soluble analogue of cyclic AMP, caused relaxation and inhibited high K(+)-stimulated Ca(2+) influx.7. Isoprenaline failed to cause stimulation of Ca(2+) efflux in high-K(+)-depolarized rabbit aorta.8. It is concluded that the inhibition of Ca(2+) influx may be one of the mechanisms by which beta-receptor stimulation can reduce intracellular free Ca(2+) to promote relaxation of smooth muscle. The data support the involvement of cyclic AMP in this action of the beta agonist.9. Since the experiments were conducted in 145 mM-K(+) (0 Na(+)) depolarizing conditions, the role of hyperpolarization or of a Na(+)-Ca(2+) exchange mechanism in isoprenaline-induced Ca(2+) influx inhibition and/or relaxation can be excluded. PMID- 6296368 TI - Blocking kinetics of the anomalous potassium rectifier of tunicate egg studied by single channel recording. AB - 1. The kinetics of the anomalous K current produced by blocking cations Na, Cs and Sr were analysed by single channel recording in the tunicate egg cell.2. The open-close kinetics in a single channel with the presence of blocking cation were consistent with the blocking kinetics of the total anomalous K current. The open close kinetics in a single anomalous K rectifier channel could be regarded as the first order transition between the open and closed states.3. The closing rate constants of the single channel for the first order transition increased almost linearly on a semilogarithmic scale as the membrane potential became more negative, while the opening rate constants decreased and then increased, showing the minimum at a certain potential level. The latter indicated that the channel was released from the blocking by excessive hyperpolarization.4. The opening rate constant increased in spite of the fixed concentration of the blocking cation (10 muM-Cs), when the external K concentration was raised from 200 mM to 400 mM. This result suggested that there are interactions within the channel between the blocking cation and the K ion.5. When there were two kinds of blocking cations such as Na and Cs, the kinetic properties in a single channel indicated that the two kinds of blocking cations blocked the single channel independently.6. The single channel conductance of the anomalous K rectifier in 200 mM-Na-containing 200 mM-K solutions was about 1.5 times larger than that in Na-free 200 mM-K solutions.7. The single channel conductances in 100, 200 and 400 mM-K solution with 10 muM-Cs were 5.0, 7.1 and 12.3 pS respectively, being roughly proportional to the square root of the K concentration.8. The density of the anomalous K rectifier channel in the tunicate egg cell was 0.039/mum(2).9. The amplitude of the single channel current increased with the rise of temperature, the Q(10) being 1.5. PMID- 6296370 TI - Comparative effects of external monovalent cations on sodium pump activity and ouabain inhibition rates in squid giant axon. AB - 1. A number of external monovalent cations were compared with regard to their effects on Na pump rate and the rate of ouabain inhibition of the pump in squid giant axon. 2. External ions which stimulate active Na efflux (K, Rb, and Cs) were found to decrease the rate at which low concentrations of ouabain inhibit the pump, and those ions which inhibit the pump externally (Na and Li) to increase the rate of inhibition. 3. In Na- and Li-containing solutions, pump rate appeared to be the major factor in determining the rate of ouabain inhibition regardless of whether K, Cs, or Rb was used to stimulate active Na efflux. 4. When choline was substituted for external Na, ouabain inhibition rates were more than twice as rapid when Cs was used as the pump-stimulating cation than when K was activating the pump to a similar level. 5. These results suggest that external monovalent cations modulate ouabain inhibition in squid axon at two classes of sites: pump activation sites, and also separate regulatory sites, whose occupation can significantly increase the rate of ouabain inhibition independent of pump turnover rate. PMID- 6296371 TI - A patch-clamp study of bovine chromaffin cells and of their sensitivity to acetylcholine. AB - 1. Bovine chromaffin cells were enzymatically isolated and kept in short term tissue culture. Their electrical properties were studied using recent advances of the patch-clamp technique (Hamill, Marty, Neher, Sakmann & Sigworth, 1981). 2. When a patch pipette was sealed tightly to a chromaffin cell ('cell-attached configuration') current wave forms due to intracellular action potentials could be observed. The frequency of the wave forms was altered by changing the pipette potential. When acetylcholine was present in the pipette solution, acetylcholine induced single channel currents were evident in the patch recording. Action potential wave forms were then often seen to follow acetycholine-induced single channel currents. 3. In the cell-attached configuration, large single channel current events did not resemble square pulses but showed exponential relaxations with time constants of the order of 50 ms. 4. After rupture of the patch of membrane, the pipette--cell seal remained stable ('whole-cell recording', Hamill et al. 1981). Chromaffin cells were found to have a resting potential of -50 to 80 mV, and an input resistance around 5 G omega. The high cell resistance accounts for the relaxing currents evident in the cell-attached configuration. 5. In the best cases, the effective time constant of the voltage clamp in the whole cell recording mode was 15 microseconds. Exchange of small ions such as Na+ ions between pipette and cell interior solutions was then complete within 15 s. 6. Acetylcholine-induced currents were obtained at various acetylcholine concentrations. Single acetylcholine-induced channels had a slope conductance of 44 pS between -100 and -55 mV, and a mean duration of 27 ms at -80 mV (at room temperature). PMID- 6296372 TI - Sodium and calcium channels in bovine chromaffin cells. AB - 1. Inward currents in chromaffin cells were studied with the patch-clamp technique (Hamill, Marty, Neher, Sakmann & Sigworth, 1981). The intracellular solution contained 120 mM-Cs(+) and 20 mM-tetraethylammonium (TEA(+)). Na(+) currents were studied after blockade of Ca(2+) channels with 1 mM-Co(2+) applied externally. Ca(2+) currents were recorded after eliminating Na(+) currents with tetrodotoxin (TTX). The current recordings were obtained in cell-attached, outside-out and whole-cell recording configurations (Hamill et al. 1981).2. Single channel measurements gave an elementary current amplitude of 1 pA at -10 mV for Na(+) channels. This amplitude increased with hyperpolarization between 10 and -40 mV, but did not vary significantly between -40 and -70 mV.3. The mean Na(+) channel open time was 1 ms at -30 mV. This open time decreased both with depolarization and hyperpolarization. Its value was close to the time constant of inactivation, tau(h), above -20 mV.4. Ensemble fluctuation analysis of Na(+) currents gave results consistent with those of single channel measurements. Noise power spectra obtained between -35 mV and 0 mV could be fitted with a single Lorentzian. A range of Na(+) channel densities of 1.5-10 channels per mum(2) was calculated.5. Cell-attached single Ca(2+) channel recordings were obtained in isotonic BaCl(2) solution. The single channel amplitude was 0.9 pA at -5 mV, and it became smaller for positive potential values.6. At -5 mV, single Ba(2+) currents appeared as bursts of 1.9 ms mean duration containing on the average 0.6 short gaps. The burst duration was larger at positive potentials.7. Ensemble fluctuation analysis of Ca(2+) channels was performed on whole-cell recordings in external solutions containing isotonic BaCl(2) or external Ca(2+) (Ca(o)) concentrations of 1 and 5 mM. The unit amplitude calculated in the former case was similar to that obtained in single channel measurements.8. Noise power spectra of Ca(2+) or Ba(2+) currents could be fitted by the sum of two, but not one, Lorentzian components.9. Tail currents could be fitted by the sum of two exponential components. The corresponding time constants had values close to those obtained with noise analysis.10. The rising phase of Ca(2+) and Ba(2+) currents was sigmoid. It could be fitted by the sum of three exponentials. The time constant of the largest amplitude component, tau(1), was similar to the time constants of the slow component observed in noise and tail experiments. This time constant also corresponded to the burst duration obtained in single channel measurements.11. The value of tau(1) was larger in 5 mM-Ca(o) and in isotonic Ba(2+) than in 5 mM-Ba(o). Thus, the kinetic properties of Ca(2+) channels depend on the nature and concentration of the permeating ion.12. A simple kinetic scheme is proposed to model the activation pathway of Ca(2+) channels.13. Currents in 1 mM-Ca(o) and 5 mM-Ca(o) showed clear reversals around +53 mV and +64 mV respectively. The outward currents observed above these potentials are most probably due to Cs(+) ions flowing through Ca(2+) channels.14. The instantaneous current-voltage relation was obtained from tail current data in the range -70 to +100 mV in 5 mM-Ca(o). The resulting curve displayed an inflexion point around the reversal potential.15. Very little inactivation of Ca(2+) currents was observed. However, a slow current decline was observed in some cells above +10 mV.16. Conditioning prepulses to positive potentials had potentiating or depressing effects on Ca(2+) currents depending on whether the test pulse lay below or above the maximal current potential. The potentiating effect may be linked to the slowest component of the current rise observed below +10 mV. The depressing effect may be related to the slow decline obtained above +10 mV.17. Analysis of ensemble variance and of tail current amplitudes suggested that the opening probability of Ca(2+) channels was at least 0.9 above +40 mV.18. A slow rundown of Ca(2+) currents was observed in whole-cell recordings. The speed of the rundown was dependent on intracellular Ca(2+) concentration. The rundown was apparently due to a progressive elimination of the channels available for activation.19. The density of Ca(2+) channels (before rundown) was estimated at 5 15/mum(2).20. In cell-attached experiments, inward current channels were often seen to follow action potentials. These events did not appear to be the usual Na(+) and Ca(2+) currents. They were probably due to cation influx of either Na(+) or Ba(2+), depending on the pipette solution, through Ca(2+)-dependent channels. Voltage-independent single channel activity observed in whole-cell and outside-out recordings may be due to the same channels. PMID- 6296373 TI - Enhanced chemosensitivity of chick parasympathetic neurones in co-culture with myotubes. AB - 1. The influence of target interaction upon the electrophysiological properties of dissociated ciliary ganglion cells was investigated by testing the sensitivity of the neuronal somal membrane to ionophoretically applied acetylcholine (ACh). Variations in the percentage of cells responsive to the transmitter were measured with time in culture. 2. Twenty-four hours after plating, all cells respond to an ionophoretic pulse of ACh with a depolarization. However, 1 week after plating (between 7 and 14 days) most of the neurones are unresponsive, and highly responsive cells (greater than 100 mV peak depolarization/nC) are extremely rare. At even later times in culture, neurones sensitive to the transmitter are again more frequent. 3. When neurones are plated onto pre-formed pectoral myotubes, however, ACh sensitivity is maintained throughout a 3 week culture period. Neuromuscular junctions are formed by the neurones, and when sufficient neurones are present, all the muscle fibres tested show evidence of functional synaptic transmission. Chemosensitivity to ACh is not maintained by neurones in muscle free microcultures are present on the same cover-slip. 4. Interneuronal synaptic contacts, defined by ultrastructural criteria, are formed in cultures of neurones alone, but evidence of widespread functional synaptic interaction between cells was not found at 7-14 days in culture. 5. It is concluded that the maintenance of ACh sensitivity of cultured ciliary ganglion cells is enhanced by the presence of muscle in co-culture. The interneuronal synaptic contacts observed are apparently not as potent a stimulus as co-culture with muscle for the full expression of the cholinergic phenotype under these culture conditions. PMID- 6296374 TI - Effect of barbiturates on the GABA receptor of cat primary afferent neurones. AB - 1. The effects of the barbiturate anaesthetics, pentobarbitone and thiopentone, on the membrane properties and the gamma-aminobutyric acid (GABA)-induced responses of cat primary afferent neurones were studied with intracellular recording and voltageclamp techniques.2. At low concentrations (10(-7)-10(-5) M) both barbiturates slightly enhanced and prolonged GABA-induced depolarizations or currents without affecting the membrane properties. At these concentrations, barbiturates have no effect on the apparent dissociation constant of the GABA GABA receptor interaction or the reversal potential for GABA-induced depolarizations or currents.3. At high concentrations (10(-4)-10(-3) M) barbiturates produced a few millivolts reduction in the resting membrane potential. Voltage-clamp analysis revealed that the depolarization was associated with one of the three types of conductance change, i.e., an initial increase followed by a decrease (40% of neurones examined), only an increase (40%) and only a decrease (20%).4. Analysis in different ionic media indicated that the depolarization with a reduced membrane resistance is associated with an increased chloride conductance and that the one with an increased membrane resistance is accompanied by a reduction in potassium conductance. Bath-application of GABA (10(-3) M) or picrotoxin (10(-5) M) inhibited the increase in chloride conductance but not the reduction in potassium conductance.5. Barbiturates at these high concentrations initially caused a marked augmentation and prolongation of GABA responses; this was followed by a depression. The depressant action did not appear to be voltage-dependent. These actions of barbiturates were not accompanied by changes in the apparent dissociation constant of the GABA-current dose-response curve or the reversal potential for GABA currents. In addition, the single exponential decay of GABA current was not changed despite a marked prolongation of its decay time.6. Picrotoxin (10(-5) M) antagonized the depressant effect of barbiturates at high concentrations on GABA currents, and barbiturates (5 x 10(-6) M) reduced the inhibitory action of picrotoxin (5 x 10( 6) M) on the GABA-currents.7. From all these results, it is suggested that the site of barbiturate actions on GABA-responses is mainly the allosteric site (the ionic conductance regulatory subunit) but not the agonist recognition site or the chloride channels linked with GABA receptors. PMID- 6296375 TI - The nature of non-cholinergic, non-adrenergic transmission in longitudinal and circular muscles of the guinea-pig ileum. AB - 1. The nature of the non-cholinergic, non-adrenergic (non-ch., non-adr.) excitatory and inhibitory transmission in the longitudinal and circular muscle layers of the guinea-pig ileum was investigated, and the effects of various agents on the junction potentials were observed using the micro-electrode method.2. In longitudinal muscle cells, ATP (3 x 10(-5)-10(-4) M) and adenosine (10(-5)-10(-4) M) depolarized the membrane, decreased the input resistance, increased the spike activity and abolished the generation of cholinergic excitatory junction potentials (e.j.p.s).3. In the presence of atropine (10(-6) M) with guanethidine (10(-5) M), field stimulation evoked three different types of the response (non-ch., non-adr. e.j.p.s, i.j.p.s (inhibitory junction potentials) or both) from cells of the longitudinal muscle layers, and only one type of the response (non-ch., non-adr. i.j.p.s) from cells of the circular muscle layer. In the following experiments atropine and guanethidine were present in the bathing fluid for at least 20 min.4. In some longitudinal muscle cells (non-ch., non-adr. i.j.p. type), ATP (5 x 10(-6)-10(-3) M) and adenosine (10(-5) 3 x 10(-5) M) depolarized the membrane, while in other cells (non-ch., non-adr. e.j.p. type), ATP (10(-5)-10(-4) M) and adenosine (10(-5)-3 x 10(-5) M) hyperpolarized the membrane and further increases in the concentration of ATP (10(-3) M) resulted in a depolarization of the membrane.5. Apamin (10(-7)-3 x 10( 6) M) inhibited the generation of non-ch., non-adr. i.j.p.s in both longitudinal and circular muscle cells, while this agent had no effect on the non-ch., non adr. e.j.p.s. As a consequence, in some cells of the longitudinal muscle layer (non-ch., non-adr. e.j.p. and i.j.p. type) the amplitude of e.j.p.s was enhanced in the presence of apamin. TEA (5 x 10(-3)-1.5 x 10(-2) M) suppressed the after hyperpolarization of the spike and i.j.p.s recorded from both muscle layers, whereas the duration and amplitudes of cholinergic and non-ch., non-adr. e.j.p.s were enhanced.6. Vasoactive intestinal polypeptide (VIP; 10(-8)-10(-7) M) had no effect on the membrane potential and junction potentials of longitudinal and circular muscle layers.7. Substance P (SP; 10(-8)-10(-7) M) depolarized the membrane of cells of the longitudinal layer (non-ch., non-adr. e.j.p. type), while this agent had no effect on cells of longitudinal (non-ch., non-adr. i.j.p. type) and circular muscle layers. SP suppressed the generation of non-ch., non adr. e.j.p.s but had no effect on i.j.p.s. Generation of non-ch., non-adr. e.j.p.s was not restored under conditions of repolarization of the membrane to the resting level by application of inward current.8. Bradykinin (BK; 10(-8)-10( 5) M) hyperpolarized the membrane and suppressed the generation of i.j.p.s in the cells of longitudinal (non-ch., non-adr. i.j.p. type) and circular muscle layers. However, when the membrane potential was displaced to the control level by outward current in the presence of BK, field stimulation evoked the i.j.p. In cells of non-ch., non-adr. e.j.p. type of the longitudinal muscle layer, BK depolarized the membrane, increased the spike activity, generated slow waves and blocked the generation of non-ch., non-adr. e.j.p.s. Displacement of the membrane potential to the control level by inward current did not restore the non-ch., non adr. e.j.p.s.9. These results suggest that in the guinea-pig ileum ATP and adenosine probably do not contribute to the generation of non-ch., non-adr. e.j.p.s and i.j.p.s, as transmitter substances. The actions of other possible candidates such as SP and BK, are discussed. PMID- 6296376 TI - Mechanisms involved in irreversible anoxic damage to the in vitro rat hippocampal slice. AB - 1. We have studied the effects of anoxia on the recovery of neural transmission between the perforant path and the dentate granule cells in the in vitro rat hippocampal slice. There is almost no recovery of the evoked population spike following 10 min of anoxia in slices from adult rats.2. A 2 h exposure of slices to creatine markedly improves the recovery of the population spike (80% vs. 5%). The creatine pre-incubation builds up phosphocreatine levels in the slice and prevents the large fall in ATP during anoxia; ATP falls to 7.9 rather than 3.6 nM/mg protein. The intracellular pH of both groups falls to the same level during anoxia.3. If calcium concentration in the medium is reduced to 0 while magnesium concentration is raised to 10 mM during anoxia the evoked response recovers to about 65%.4. The data suggest that an attenuation of the fall in ATP or entry of calcium during anoxia protects the tissue against irreversible transmission damage. Thus both of these factors participate in the generation of this damage. It is not yet clear if they act independently or if one acts by altering the other.5. In the post-anoxic recovery period the intracellular concentration of potassium is reduced by about 25%. However, it is still much higher than in slices that show only partial block of the evoked response when treated with ouabain. Therefore a fall in intracellular potassium 1 h after anoxia cannot explain the lack of recovery of the evoked response in adult tissue.6. ATP levels in the post-anoxic recovery period are reduced from their pre-anoxic levels (9.7 vs. 13.9 nM/mg protein). However when azide or antimycin A are used to directly reduce ATP to the level found 1 h after anoxia the evoked response is reduced by only about 45%. Thus the reduced post-anoxic ATP levels are not sufficient to explain the loss of the evoked response in adult tissue.7. The data show that the irreversible loss of transmission is not due to decreased cell ATP or to decreased cell K/Na levels 1 h after the anoxic period.8. Since creatine pre incubation protects against irreversible transmission loss this compound or one closely related to it may prove useful in attenuating irreversible brain damage in situ. PMID- 6296377 TI - Cholinergic transmission in cat parasympathetic ganglia. AB - 1. Intracellular electrical recording techniques were used to study the ionic mechanisms of cholinergic synaptic transmission in cat vesical pelvic ganglia (v.p.g.). 2. Orthodromic nerve stimulation as well as ionophoretic application of acetylcholine (ACh) resulted in, first, a fast excitatory post-synaptic potential (f.e.p.s.p.) and secondly, a slow inhibitory post-synaptic potential (s.i.p.s.p). These distinct post-synaptic responses were direct actions of ACh and not mediated through an interneurone. In addition, a slow excitatory post-synaptic potential (s.e.p.s.p.) was observed in 44% of the cells. 3. The f.e.p.s.p., mediated via nicotinic receptors, had a reversal potential of -10 mV and resembled the conventional rapid depolarization in other ganglia. The s.i.p.s.p., mediated by muscarinic receptors, had a reversal potential of about -100 mV and resulted from an increase in potassium conductance. 4. The slow muscarinic hyperpolarization could be observed in the absence of antagonists and it was elicited at stimulus frequencies in the physiological range (2-10 Hz). the s.i.p.s.p. induced orthodromically or ionophoretically inhibited firing in spontaneously active neurones. These observations suggest that the muscarinic hyperpolarization may occur under physiological conditions and has sufficient magnitude to be inhibitory to neuronal activity. PMID- 6296378 TI - Transmucosal implants of dense hydroxylapatite. AB - Transmucosal application of implants of dense hydroxylapatite has been studied in a long-term animal experiment. Implants were loaded with crowns or removable restorations. It appeared that transmucosal implants adhered to bone similarly as in the submucosal application. The clinical aspect of gingival tissues around the implants was the same as that around the adjacent natural teeth. Histologically, there were indications for a good attachment of gingival tissues to the implants. Implants fractured due to fatigue failure of the ceramic material. Prestressing of the implants may be the solution to this problem. Failing implants can clinically be treated by submerging, removal, or substitution. PMID- 6296379 TI - [Radiological aspects of malignant histiocytic pathology]. AB - The histiocyte, macrophage of tissue monocyte is a cell of the mononuclear system. At the moment several different types of classification of the malignant histiocytic pathology are use. Seven cases of this pathology are documented: two cases of malignant histiocytosis, two cases of malignant histiocytic lymphoma, two cases of malignant histiocytoma of soft tissue and one malignant histiocytic tumor of bone. The purpose of this study is to analyse the usefulness the roentgenologic methods: conventional radiography, radionuclide scanning, computed tomography, sonography, xeroradiography, angiography. The evaluation of malignant histiocytic pathology has become more precise through the introduction of ultrasonography and computed tomography. This report adds to the understanding of the entities of this pathology in that the diagnosis of malignance was made using roentgenologic criteria and confirmed by histology. PMID- 6296380 TI - Isolation and partial characterization of an acetylcholine receptor-enriched membrane fraction from skeletal muscle. AB - A procedure for purification of the bungarotoxin-binding fraction of sarcolemma from rabbit skeletal muscle is described. Muscle is homogenized in 0.25M sucrose without high salt extraction and membrane fractions separated initially by differential centrifugation procedures. An ultracentrifugation pellet enriched in cell surface and sarcoplasmic reticulum markers is further fractionated on a dextran gradient (density = 1.0 to 1.09). Two fractions are identified as sarcolemma according to high specific activities for lactoperoxidase-iodination, Na+, K+-ATPase and alpha-bungarotoxin-binding. No Ca++, Mg++-ATPase activity is found in these fractions. A third fraction, the dextran gradient pellet, is enriched in Ca++, Mg++-ATPase activity and lactoperoxidase iodinatable material and characterized by low bungarotoxin binding. This fraction represents a mixture of sarcoplasmic reticulum and transverse tubules with some sarcolemma contamination. PMID- 6296381 TI - Effects of temperature, nucleotides and sodium molybdate on activation and DNA binding of estrogen, glucocorticoid, progesterone and androgen receptors in MCF-7 human cancer cells. AB - We studied the effects of temperature, ribonucleotides and sodium molybdate on the activation and DNA cellulose binding of estrogen, glucocorticoid, progesterone and androgen receptor complexes in MCF-7 cells. Using DNA cellulose binding as a measure of receptor activation, we found that ribonucleotides activated all four of these receptor complexes. Temperature also activated glucocorticoid receptor complexes efficiently but activated progesterone and androgen receptor complexes less well. Temperature did not activate estrogen receptor complexes. Sodium molybdate blocked either ATP or temperature induced activation of glucocorticoid, progesterone and androgen receptor complexes but only partially blocked estrogen activation. Sodium molybdate also prevented the formation of multiple forms of estrogen and glucocorticoid receptor complexes seen on DEAE cellulose and hydroxylapatite chromatography of crude cytosol. The mechanism by which ribonucleotide enhances and molybdate inhibits activation are discussed. PMID- 6296382 TI - Characterization of sialoglycoproteins of rat epididymal fluid and spermatozoa by periodate-tritiated borohydride. AB - Sialoglycoproteins of rat epididymal fluid and spermatozoa were radiolabelled by the NaIO4/KB3H4 method. At least 10 sialoglycoproteins of the epididymal fluids could be consistently demonstrated by polyacrylamide gel electrophoresis in sodium dodecyl sulphate. Two major ones (Mr 21000 and 66000) were present in the fluids of the caput and cauda epididymidis. Two (Mr 28000 and 40000) were found only in the former and two (Mr 32000 and 42000) only in the latter. There were at least 11 sialoglycoproteins bound to the epididymal spermatozoa. During epididymal transport, 8 sialoglycoproteins on the spermatozoa decreased, one (Mr 48000) remained constant and one (Mr 31000) increased in amount. During sperm maturation, some sperm-bound sialoglycoproteins, especially 3 of small molecular weights, became more resistant to the treatments with neuraminidase, trypsin and Triton X-100. PMID- 6296383 TI - Antiserum to LH reverses the abortifacient effect of Bromergocryptine treatment in early rat pregnancy. AB - In vivo experiments showed that anti-bovine luteinizing hormone rabbit serum (a LH) reversed the abortifacient effect of Bromergocryptine (BEC) on day 6 of pregnancy in 13 of 19 rats when injected 5-15 h after the initial BEC administration. Radioreceptor assay studies revealed that BEC treatment entailed an 85% drop of ovarian LH-receptor (LH-rec) concentration and a drastic decrease in progesterone serum levels. a-LH restored both the ovarian LH-rec content and progesterone serum levels in those rats that were found to have macroscopically intact implantations. A time course study of LH serum levels after BEC treatment showed a trend for BEC to stimulate LH secretion with a maximum 12-18 h after the initial BEC injection. These experiments indicate that (i) BEC terminates early rat pregnancy primarily by inducing a process of desensitization of ovarian LH rec by blocking prolactin (PRL) secretion and possibly also by additionally stimulating LH secretion, which in the absence of endogenous PRL causes HL-rec and serum progesterone levels to drop; (ii) a-LH administration without simultaneous PRL substitution may rescue LH-rec of BEC treated rats from being desensitized as well as prevent fetuses from being resorbed. It is suggested that the contribution of PRL in the luteotropic complex is permissive for the luteotropic action of LH and/or for keeping the corpus luteum in a functional state rather than being luteotropic by itself. PMID- 6296384 TI - Breast screening in the primary care office. A plea for early detection. PMID- 6296385 TI - Characterization of nonlymphoid cells in rat spleen, with special reference to strongly Ia-positive branched cells in T-cell areas. AB - By use of a monoclonal antibody against Ia antigen in an immunoperoxidase method, strongly Ia-positive branched cells are found in the T-cell areas of the splenic white pulp of the rat. In order to further characterize these cells, enzyme histochemical characteristics, phagocytic capacity, and irradiation sensitivity have been studied. Evidence is presented that these strongly Ia-positive branched cells represent interdigitating cells. The influence of whole-body irradiation on interdigitating cells is discussed. Comparison with data from the literature on the in vitro dendritic cell isolated from spleen cell suspensions reveals many similarities between the described interdigitating cell in vivo and the dendritic cell in vitro. PMID- 6296386 TI - Changes in cAMP metabolism during phagocytosis of S. aureus by human monocytes. AB - Involvement of cyclic adenosine 3',5'-mono-phosphate (cAMP) in the phagocytosis of staphylococci by human monocytes was demonstrated by assay of adenylate cyclase and cAMP phosphodiesterase (PDE). Monocyte adenylate cyclase and PDE activities were assayed on cell homogenates prepared from monocytes isolated by plate adherence. Phagocytosis of staphylococci was associated with diminished adenylate cyclase activity, which reached a minimum after 10 min of incubation (p less than 0.05), and an increase in PDE activity (p less than 0.005). Transmission electron microscopy provided evidence that these changes coincide with peak phagocytic activity occurring between 10 and 20 min after the start of phagocytosis but could not be assigned to a particular stage in the phagocytic process. PMID- 6296387 TI - alpha 2 adrenoceptors: classification, localization, mechanisms, and targets for drugs. PMID- 6296388 TI - Synthesis and pharmacological characterization in vitro of cyclic enkephalin analogues: effect of conformational constraints on opiate receptor selectivity. AB - Using a combination of solid-phase and solution methods, we synthesized a series of cyclic [Leu5]enkephalin analogues by substitution of D-alpha, omega-diamino acids in position 2 of the enkephalin sequence and cyclization of the omega-amino group to the C-terminal carboxy group of leucine. Cyclic analogues containing D alpha, beta-diaminopropionic acid (1), D-alpha, gamma-diaminobutyric acid (2), D ornithine (3), or D-lysine (4) in position 2 and the [D-Leu5] and [des-Leu5] analogues of 4 (5 and 6) showed, in general, high potency in the guinea pig ileum (GPI) assay and low potency in the mouse vas deferens (MVD) assay. IC50 (MVD)/IC50 (GPI) ratios ranging from 3.1 to 29.4 were obtained, indicating the preference of the cyclic analogues for mu receptors over delta receptors. With two exceptions, preferential affinity for mu receptors is reflected in the Ki ratios determined in parallel binding assays using [3H]naloxone and [3H] [D-Ala2, D-Leu5]enkephalin as mu and delta receptor selective radioligands, respectively. Comparison of the pharmacological profiles of the cyclic analogues 1-4 with those of their corresponding open-chain analogues, [D-Ala2, Leu5]enkephalinamide (1a), [D-Abu2, Leu5]enkephalinamide (2a), [D-Nva2, Leu5]enkephalinamide (3a), and [D Nle2, Leu5]enkephalinamide (4a), revealed that the pronounced mu character of compounds 1-4 is a direct consequence of the conformational constraints introduced by cyclization. This finding is in agreement with the concept of different conformational requirements of mu- and delta-opiate receptors and raises the possibility of manipulating opiate receptor selectivity by varying the type and degree of conformational restriction. PMID- 6296390 TI - Crystal structure and anti herpes simplex virus activity of 2,2'-anhydro-1-beta-D arabinofuranosylthymine. AB - 1-beta-D-Arabinofuranosylthymine (aThy; ara-T) is a potent selective anti herpes simplex virus drug. Its anhydro analogue, 2,2'-anhydro-aThy, was shown to be 9 fold less active and at least 3-fold less toxic than aThy. This compound was relatively stable at physiological pH and in strong acid but was rapidly hydrolyzed in base with a half-life of 18.3 min. The three-dimensional crystal structure of 2,2'-anhydro-aThy revealed a rigid structure with the arabinose ring in the unusual O1' endo, pucker, conformation. The trans-gauche conformation along the C4'-C5' bond permits only intermolecular hydrogen bonding of the 5' hydroxy and O3'. PMID- 6296389 TI - 3'-Hydroxy- and (+/-)-3',11-dihydroxy-delta 9-tetrahydrocannabinol: biologically active metabolites of delta 9-tetrahydrocannabinol. AB - Synthesis of 3'-hydroxy- (7) and (+/-)-3',11-dihydroxy-delta 9 tetrahydrocannabinol (11), metabolites of delta 9-THC, is described. Condensation of the monoterpene (+/-)-cis-p-menth-2-ene-1,8-diol (1) and (+/-)-3' acetoxyolivetol (2) in the presence of fused ZnCl2 in CH2Cl2 gave a mixture from which the delta 9-THC derivative 3, containing small amounts of the delta 8 isomer 4, was isolated after column chromatography. This mixture was separated as their diacetates 5 and 6 by high-pressure liquid chromatography. Alkaline hydrolysis (5% KOH in MeOH) of 5 furnished the metabolite 7. Condensation of (+/ )-8 with 2 in the presence of p-toluenesulfonic acid gave 9a, which was acetylated to 9b. Treatment with HgO/BF3 X Et2O in wet THF gave the aldehyde 10. Reduction with LiAlH4 furnished the metabolite 11. These metabolites were compared with delta 9-THC for their ability to depress spontaneous activity and rectal temperature in mice and for their effects on overt behavior in dogs. 3' Hydroxy-delta 9-THC was also compared to delta 9-THC in the mouse sympatomatology test and cardiovascular system in dogs. The metabolites produced pharmacological effects similar to those of delta 9-THC in all tests. 3'-Hydroxy-delta 9-THC was 2-3 times more effective than delta 9-THC in the behavioral tests, whereas (+/-) 3',11-dihydroxy-delta 9-THC was approximately 3 times less active than delta 9 THC. PMID- 6296392 TI - How macrophages kill tubercle bacilli. PMID- 6296391 TI - Genetics of plasma paroxonase activity. PMID- 6296393 TI - A sequence of the yeast 2 micron DNA plasmid chromosome near the origin of replication is exposed to restriction endonuclease digestion. PMID- 6296394 TI - Plasmid R6K DNA replication. II. Direct nucleotide sequence repeats are required for an active gamma-origin. PMID- 6296395 TI - Electron microscopic analysis of ts1 10 Moloney mouse sarcoma virus, a variant of wild-type virus with two RNAs containing large deletions. PMID- 6296396 TI - Structural organization of alpha-satellite DNA in a single monkey chromosome. PMID- 6296397 TI - Isolation and structural analysis of ribosomal protein genes in Xenopus laevis. Homology between sequences present in the gene and in several different messenger RNAs. PMID- 6296398 TI - Formation of composite iso-cytochromes c by recombination between non-allelic genes of yeast. PMID- 6296400 TI - Precise nucleotide location of the 5' ends of RNA-primed nascent light strands of mouse mitochondrial DNA. PMID- 6296399 TI - Non-contiguous segments of the polyoma genome required in cis for DNA replication. AB - The boundaries of the origin of polyoma DNA replication have been analyzed using a set of deletion mutants. The majority of these had small deletions, 5 to 30 basepairs in size, which together removed most of the non-translated sequences of the genome. The phenotype of the mutants was characterized by analysis of infectivity, transforming ability and DNA synthesis. All mutants with reduced or abolished infectivity had corresponding defects of viral DNA synthesis. The effect of the deletion was cis-acting, since the replication of the mutants was not stimulated by the presence of wild-type DNA. Deletions causing a reduction of DNA synthesis were found at two sites. The first at the 32 base-pair inverted repeat sequence and the neighbouring A . T tract previously implicated in the initiation of DNA synthesis, and the second close to the late genes. The two sites were separated by at least 60 base-pairs of non-essential DNA. Only one mutant with a deletion at the second site was unable to express early gene functions. The mutants were constructed by linearization, shortening and recircularization of polyoma DNA inserted into the plasmid pBR322. The mutagenesis was directed at restriction endonuclease BglI or PvuII cleavage sites. The BglI-directed mutagenesis was focussed to polyoma DNA by using as a vector a derivative of pBR322 resistant to cleavage by BglI. PMID- 6296401 TI - Structure of the cytochrome c oxidase dimer. Electron microscopy of two dimensional crystals. PMID- 6296402 TI - Membranous cytochrome c oxidase. A freeze-fracture electron microscopic analysis. PMID- 6296403 TI - R-prime site-directed transposon Tn7 mutagenesis of the photosynthetic apparatus in Rhodopseudomonas capsulata. PMID- 6296404 TI - A competing salt-bridge suppresses helix formation by the isolated C-peptide carboxylate of ribonuclease A. PMID- 6296405 TI - Oligomerization of (guanosine 5'-phosphor)-2-methylimidazolide on poly(C). An RNA polymerase model. PMID- 6296406 TI - Infrared study of G . C complex formation in template-dependent oligo(G) synthesis. PMID- 6296408 TI - Perfluorochemical perfusion of the rat heart: in vivo and in vitro. PMID- 6296407 TI - Expression of the hepatitis B virus surface, core and E antigen genes by stable rat and mouse cell lines. PMID- 6296410 TI - Slow inactivation of calcium channels in the cardiac Purkinje fiber. PMID- 6296409 TI - Loss of the cyclic AMP accumulation induced by isoproterenol during acute ischaemia in the isolated rat heart. PMID- 6296411 TI - Ouabain-induced tachyarrhythmias and cell damage in isolated perfused guinea-pig hearts: I. Protection by propranolol. PMID- 6296412 TI - Propranolol in ischaemic reperfused hearts. PMID- 6296413 TI - Increase of a nerve growth factor-like protein in the cerebellum of PCD mutant mice. PMID- 6296414 TI - Nerve growth factor-induced stimulation of alpha-aminoisobutyric acid uptake in PC12 cells: relationship to plasma membrane receptor occupancy. AB - Nerve growth factor (NGF) stimulates the uptake rate of the nonmetabolized amino acid alpha-aminoisobutyric acid (AIB) in the clonal PC12 pheochromocytoma cell line by 40-70%. This effect reaches a maximum after a 1-hour incubation with the hormone and then drops over 50%, reaching a minimum after 4 hours of NGF administration. Longer exposure to the hormone leads to a gradual rise in stimulation, and by 24 hours, the cells regain about 80% of the original 1-hour rate. Results of NGF-binding studies indicate that stimulation of AIB uptake follows closely behind the amount of NGF bound to the low-affinity NGF receptors. Dose-response experiments indicate that full stimulation occurs biphasically. Within the NGF concentration range of 0.1 ng/ml to 1 ng/ml, the AIB uptake rate is 30% of the maximum 1-hour response. At 2.5 ng/ml NGF, the stimulation jumps to about 75% maximal response while 100% response is reached between 5 ng/ml and 50 ng/ml NGF. We suggest that the two states of the NGF plasma membrane receptor on PC12 cells may both be involved in mediating NGF stimulation of AIB uptake. PMID- 6296415 TI - Nerve growth factor effects and receptors in cultured human neuroblastoma cell lines. AB - We studied the effects of nerve growth factor (NGF) to determine whether neuroblastoma (NB) cells share the pattern of altered response to growth regulatory factors shown by various malignant transformed cells. NGF induces neurite outgrowth, arrests growth, and enhances survival in normal neurons and in the rat pheochromocytoma, a tumor cell closely related to NB. With respect to neurite outgrowth, lines SK-N-SH, SH-SY5Y, LA-N-5, and CHP-126 were sensitive, IMR-32 was resistant, and SH-EP1, SK-N-MC, MC-IXC, CHP-100, and CHP-134 were unresponsive. Conditioned media from unresponsive cells did not inhibit response in sensitive cells. Unexpectedly, NGF neither reduced the growth rate nor enhanced survival in any NB cell line. Conditioned medium from all NB cell lines enhanced 125I-NGF binding in embryonic sensory cells. Regulation of growth rate and neurite outgrowth, then, are separable. A fundamental defect in NB may be the acquisition of a capacity for growth and survival independent of NGF. 125I-NGF was bound to both Fast and Slow receptors in MC-IXC cells, but only to Slow receptors in NGF-responsive SH-SY5Y and LA-N-5 cells, showing Fast receptors are not required for neurite outgrowth. Independence from NGF-regulated growth and survival is unexplainable by an absence of NGF receptors. PMID- 6296416 TI - Nerve growth factor and cyclic AMP: opposite effects on neuroblastoma-substrate adhesion. AB - Increased cellular adhesion has been postulated to be an early event required of neuroblasts undergoing neurite extension during differentiation. Nerve growth factor (NGF) induces neurite extension in a variety of cell types of neural crest origin. In the PC12 rat pheochromocytoma cell line, which has been proposed as a model for precursor cells to sympathetic neurons, NGF and dibutyryl cyclic AMP (dBcAMP) promote both neurite extension and an increased rate of cell-substrate adhesion. Since dBcAMP can substitute for NGF in this enhancement of adhesion rate in the PC12 cell line, cAMP has been suggested as a second messenger for NGF. We have shown that in two nearly diploid adrenergic like human neuroblastoma clones, the KA and SY5Y cell lines, which also extend neurites in response to both NGF and dBcAMP, only NGF enhances cellular adhesion, as defined by an increase in the number of cells cells prelabeled with 35S-methionine which attach to culture dishes at a given time. Incubation with monospecific antibodies directed against murine beta-NGF abolishes the NGF effect on adhesion. The NGF effect on human neuroblastoma is specific insofar as NGF does not facilitate adhesion of two glioma lines. Unlike the results obtained for PC12, in both SY5Y and KA lines, 1 mM dBcAMP decreases the rate of adhesion to levels significantly below those of controls. Adhesiveness of neuroblastoma cells treated with both NGF and dBcAMP is intermediate between that of cells treated with either agent alone. While theophylline mimics the dBcAMP effect, sodium butyrate has no such effect. At 22 degrees C, the effect of NGF on neuroblastoma substrate adhesion is observable within 5 minutes and persists for 2 hours; treatment of the KA line with NGF at 37 degrees C for 24 hours results in a more persistent enhancement of cell adhesion. Furthermore, both SY5Y and KA exhibit different morphologies when challenged with NGF, dBcAMP, or sodium butyrate. This study suggests that NGF and cyclic AMP do not share a common mechanism of action, but can in fact interact antagonistically in an adrenergic like neuroblastoma model system. Furthermore, the results suggest that increased cellular adhesiveness may not be an obligatory prerequisite for neurite extension by neuroblasts in development. PMID- 6296417 TI - Epidermal growth factor binding and mitogenic activity on purified populations of cells from the central nervous system. AB - Binding of 125I-epidermal growth factor (EGF) to purified populations of rat astrocytes, oligodendrocytes, and neurons was measured. Astrocytes bound 40,000 100,000 EGF molecules per cell, while oligodendrocytes bound only 6,000-10,000 EGF molecules per cell. In contrast, neurons had little or no capacity to bind 125I-EGF. EGF alone was able to stimulate incorporation of tritiated thymidine fivefold in purified astrocyte cultures incubated in serum-free medium. When EGF was added to the previously described chemically defined medium for astrocytes, incorporation of tritiated thymidine in purified astrocytes was equivalent to that observed in medium containing 10% fetal calf serum. Addition of EGF to this chemically defined medium also doubled the proliferative capability of the medium for cultured astrocytes. EGF was maximally effective in stimulating 3H-thymidine incorporation at concentrations between 1-10 ng/ml. PMID- 6296418 TI - Effect of epidermal growth factor on membrane motility and cell locomotion in cultures of human clonal glioma cells. AB - Two clones, designated Cl 2 and Cl 3, were established from the human malignant glioma line U-343 MGa. The astrocytic origin of the cells was proven by the presence in virtually 100% of the cells of the astrocyte marker glial fibrillary acidic protein. The addition of 10 ng epidermal growth factor (EGF) per milliliter to Cl 2 and Cl 3 cells resulted in the rapid appearance of large cell surface ruffles, visualized by scanning electron microscopy. A time course study by phase contrast microscopy showed that the maximal ruffling activity occurred 5 minutes after addition of EGF. Under basic culture conditions (Eagle's MEM, 10% fetal calf serum), Cl 2 and Cl 3 cells were essentially immobile and formed tightly packed, well demarcated colonies. In the presence of 10 ng EGF per milliliter, no defined colonies were formed and the cells seemed to move around freely. The stimulatory effect of EGF on cell migration was confirmed by growing the cells on a deposit of colloidal gold; in the absence of EGF, the cells remained immobile whereas cells grown at 10 ng EGF per ml formed long phagokinetic tracks. The effect of EGF on membrane motility and cell locomotion occurred in the absence of any effect of EGF on growth rate; both clones multiplied at the same rate in the absence as in the presence of EGF. Binding experiments using 125I-labeled EGF demonstrated a single class of high affinity receptors. The number of 180,000 receptors per cell was estimated in both clones. The finding that human glioma cells in culture require EGF for their migration raises the interesting possibility that tumor cells in vivo may respond in a similar fashion, and in that case require a growth factor for migration and for the expression of their infiltrative growth potential. Furthermore, the present findings strengthen the notion that glial cells should be recognized as targets for EGF. PMID- 6296419 TI - Biochemical aspects of pentylenetetrazole induced seizure. AB - At the onset of pentylenetetrazole induced convulsions, the adenylate cyclase activity and phosphodiesterase activity were increased. The former was markedly stimulated in the brain stem of rats. In the cerebral cortex and brain stem, the glucose level was significantly decreased, and the concentration of glucose-6 phosphate was increased. However, the definite changes in energy reserve system of the brain could not be observed at the onset of penetylentetrazole induced seizures. The present study revealed some correlation between pentylenetetrazole convulsions and the adenylate cyclase activity and glycometabolism. PMID- 6296420 TI - Computed tomography and angiofibroma of the nasopharynx. PMID- 6296421 TI - Effects of sodium acetate, bicarbonate and lactate on acid-base status in anaesthetized dogs. AB - Equal doses of sodium acetate, bicarbonate and lactate (6.6 mEq/L) infused intravenously over 30 min into three groups of halothane-anaesthetized dogs caused changes in acid-base status. Arterial carbon dioxide tension (PaCO2), pHa, base excess (BE) and standard bicarbonate (SB) increased. Sodium bicarbonate caused the most rapid and greatest changes. The bicarbonate group was significantly different (P less than 0.05) from the other groups at 15 and 30 min after the start of infusion for pHa, BE and SB. The greater effects of bicarbonate are due to its production of alkalinization without a requirement for metabolism; acetate and lactate require oxidation to be effective. The acetate and bicarbonate groups were not statistically different at 45 min after the onset of drug infusion, but both had significantly higher SB and BE mean values than the lactate group. All measurements made after 45 min revealed no significant differences among groups. Thus, after the earlier differences noted above, the three alkalinizers caused similar effects on acid-base status. PaCO2 was elevated in all groups, but there were no differences among groups. Cardiovascular effects caused by infusion of the three drugs were minimal. PMID- 6296422 TI - Trimetaquinol relaxes bovine pulmonary arteries in vitro through a non-beta adrenergic mechanism. PMID- 6296423 TI - Nucleotide sequence of the envelope gene of Friend murine leukemia virus. AB - The envelope gene of the helper-independent, highly leukemogenic virus Friend murine leukemia virus was sequenced by using a molecular clone of a Friend murine leukemia provirus. The deduced amino acid sequences of the envelope proteins gp70 and p15env were homologous to the sequences of Moloney murine leukemia virus (86%) and Akv (76%). However, a stretch of about 40 amino acid residues near the middle of gp70 was dissimilar in Friend and Moloney murine leukemia viruses and Akv. In this type-specific region the gp70s of all three viruses contained more than 30% proline residues, giving this sequence a very rigid conformation. We suggest that this rigid and highly variable region of gp70 participates in infection by recognition of cell surface receptors and, in addition, might contribute to the different oncogenic spectra of murine leukemia viruses. PMID- 6296424 TI - Transcription of herpes simplex virus genes in vivo: overlap of a late promoter with the 3' end of the early thymidine kinase gene. AB - We identified in herpes simplex virus type 1-infected cells six cytoplasmic transcripts which were complementary to BamHI restriction endonuclease fragment Q. Two transcripts appeared in major amounts compared with the other four. One major transcript of about 1.4 kilobases was the mRNA for the viral thymidine kinase, was synthesized at intermediate times, and was classified as a beta transcript. The other major transcript was synthesized at late times and was classified as a gamma transcript. This late transcript was about 3 kilonucleotides long and was transcribed in the same direction as the gene for thymidine kinase. The 5' end of this late RNA was located by RNA sequence analysis and was 23 nucleotides downstream from the polyadenylation site for the thymidine kinase mRNA. This finding led to the conclusion that the control region for the 3-kilobase gamma transcript is contained within the 3' untranslated region of the thymidine kinase transcript. PMID- 6296425 TI - Structure and expression of c-rel, the cellular homolog to the oncogene of reticuloendotheliosis virus strain T. AB - The cellular homolog of the onc sequences in the avian retrovirus reticuloendotheliosis virus strain T (v-rel) was studied by molecular cloning and nucleic acid hybridization. In contrast to v-rel sequences, which are 1.4 kilobase pairs long, the cellular homolog, c-rel, from line 15B chickens is at least 25 kilobase pairs long, with multiple apparent introns. A 4.0-kilobase polyadenylic acid-containing RNA transcript is the primary species of c-rel RNA present in uninfected chicken cells. Sequences at the 3' end of this c-rel RNA are not present in v-rel. PMID- 6296426 TI - Integration of new endogenous mouse mammary tumor virus proviral DNA at common sites in the DNA of mammary tumors of C3Hf mice and hypomethylation of the endogenous mouse mammary tumor virus proviral DNA in C3Hf mammary tumors and spleens. AB - To understand the molecular mechanisms by which the endogenous murine mammary tumor virus (MuMTV) proviruses are expressed and produce late-occurring mammary tumors in C3Hf mice, we analyzed, by the use of restriction enzymes and the Southern transfer procedure, genomic DNA from normal organs of mammary tumor bearing and tumor-free mice and from 12 late-occurring C3Hf mammary tumors. We found, by using the restriction enzymes EcoRI and HindIII, that in addition to the preexisting endogenous MuMTV proviruses, new MuMTV-specific proviral DNA was integrated into new sites in the host genome in all 12 of the tumors that we examined. PstI digests of C3Hf tumor DNA revealed that the new proviral DNA found in C3Hf tumors was of endogenous origin. Moreover, the respective sizes of at least one of the new DNA fragments generated by EcoRI or HindIII digestion were the same in at least 50% of the C3Hf tumors analyzed, suggesting that the integration site of this new proviral DNA could be at the same location in the host genome of these tumors. Our results may imply that mammary tumorigenesis in C3Hf mice results from activation of cellular oncogenes by an MuMTV proviral DNA promoter. Specific hypomethylation of MuMTV proviral DNA was detected in the mammary tumors and spleens of C3Hf tumor-bearing mice. Our results indicated that most, if not all, of the hypomethylated MuMTV proviral DNA sequences were derived from the endogenous MuMTV provirus located at the MTV-1 locus, a locus responsible for the production of MuMTV antigens and increased incidence of mammary carcinoma in C3Hf mice. In spleens of non-tumor-bearing mice of ages 3, 6, 9, and 12 months, there was progressive hypomethylation of proviral DNA with increasing age, suggesting a possible correlation between demethylation of MuMTV proviral DNA in the spleens of C3Hf mice and the expression of endogenous MuMTV. PMID- 6296427 TI - RNA and protein encoded by MH2 virus: evidence for subgenomic expression of v myc. AB - MH2 and MC29 are highly related myc-containing avian retroviruses. We found that MH2, unlike MC29, synthesizes a 2.6-kilobase subgenomic mRNA containing myc sequences as well as sequences from the 5' end of the genome. A 57-kilodalton protein containing myc, but not gag, sequences (p57myc) was detected by hybrid selection and in vitro translation of RNA from MH2-transformed cells. Gradient separation of MH2 intracellular RNAs indicated that p57myc is encoded by the subgenomic RNA. A highly oncogenic MH2 virus variant (MH2YS3) (M. Linial, Virology 119:382-391, 1982) was shown to encode only p57myc and not P100, the previously described MH2-encoded polyprotein (Hu et al., Virology, 89:162-178, 1978). Cells transformed by subclones of this virus synthesized predominantly the 2.6-kilobase RNA rather than genomic 5.4-kilobase RNA. These results suggest that only p57myc is required for maintenance of the transformed state after MH2 infection. PMID- 6296428 TI - Identification of a cellular receptor for mouse mammary tumor virus and mapping of its gene to chromosome 16. AB - Pseudotypes of vesicular stomatitis virus (VSV) containing envelope glycoproteins provided by C3H mammary tumor virus (MTV) instead of the normal VSV G-proteins were prepared and used to assay the presence of an MTV receptor on cells. The assay was specific as demonstrated by competition studies with excess MTV particles and neutralization of the pseudotypes with anti-MTV serum or monoclonal antibodies directed against MTV gp52. The MTV receptor was abundantly present on mouse cells but hardly detectable on nonmurine cells, including the Chinese hamster cell line E36. Somatic cell hybrids between E36 cells and GRS/A spontaneous leukemia cells (GRSL cells) and between E36 and GRS/A primary mammary tumor cells were made. The hybrids retained all Chinese hamster chromosomes but segregated mouse chromosomes. From the analysis of the isoenzymes and chromosomes of the hybrid cell lines we conclude that the gene for the receptor (MTVR-1) is located on mouse chromosome 16. PMID- 6296429 TI - Cloned mouse mammary tumor virus DNA exhibits glucocorticoid-dependent expression in simian virus 40-transformed mink cells. AB - Mink lung epithelial cells were transfected with two cloned mouse mammary tumor virus (MMTV) DNAs, a 9-kilobase clone derived from an unintegrated exogenous viral genome and a 14-kilobase clone containing an integrated endogenous provirus along with cellular flanking sequences. Mink lung cells were chosen because they do not contain endogenous MMTV sequences. On the basis of our observation that simian virus 40 DNA efficiently transforms these cells, we isolated cell clones containing MMTV DNA by using transformation with simian virus 40 DNA as a selective marker in cotransfection experiments. Levels of the 9-kilobase MMTV mRNA representing the entire viral genome and of the spliced 4.4-kilobase mRNA which codes for the viral envelope proteins were glucocorticoid dependent in transformed cells. Expression of low levels of Pr77gag, the precursor of the group-specific viral core proteins, and of gPr73env, the precursor of the viral envelope proteins, was also hormone dependent. We conclude that these cloned MMTV DNAs contain all the information necessary for the synthesis of normal viral RNAs and proteins. These findings also provide further evidence that the DNA sequences involved in the hormone responsiveness of MMTV expression are contained within the viral genome. PMID- 6296430 TI - Cell-free synthesis and assembly of vesicular stomatitis virus nucleocapsids. AB - The association of newly synthesized vesicular stomatitis virus proteins into nucleocapsid structures was examined in a cell-free system that supports concurrent viral protein synthesis, transcription, and RNA replication. The vesicular stomatitis virus proteins synthesized by this system associated with the newly replicated RNA to form structures that banded in CsCl gradients with marker nucleocapsids. In reactions lacking nucleocapsid templates to program RNA synthesis, the newly synthesized proteins did not associate into nucleocapsid structures. The newly synthesized proteins associated with nucleocapsids were analyzed by electrophoresis on polyacrylamide gels containing sodium dodecyl sulfate after separation from non-associated proteins by chromatography on Bio Gel A15M agarose columns. The results of this analysis showed that newly synthesized L, NS, and N proteins associated into nucleocapsids in the in vitro system. In addition, a small amount of newly synthesized M protein was stably bound to the nucleocapsids. The molar ratio of the associated, newly synthesized proteins was 2:350:1,000:10 (L:NS:N:M). More than 90% of the newly synthesized NS protein that associated with nucleocapsids in vitro was of the NS2 subspecies, as assayed by DEAE-cellulose column chromatography. The stability of the association of the newly synthesized proteins with nucleocapsids in the system mimicked that of the association of viral proteins with nucleocapsids from infected cells as measured by salt sensitivity. These data indicate that nucleocapsids were assembled from newly synthesized proteins within our in vitro system and that the molar ratio of assembled proteins was similar to that observed for virion nucleocapsids. PMID- 6296431 TI - Effect of intracellular vesicular stomatitis virus mRNA concentration on the inhibition of host cell protein synthesis. AB - Inhibition of host cellular protein synthesis by vesicular stomatitis virus (VSV) has been suggested to be primarily the result of competition for ribosomes between cellular and viral mRNAs (H. F. Lodish and M. Porter, J. Virol., 36:719 733, 1980; Lodish and Porter, J. Virol. 38:504-517, 1981). This hypothesis was investigated by regulating the extent of VSV mRNA synthesis through the use of defective interfering particles. Although intracellular VSV mRNA concentrations decreased by as much as a factor of 14 at high multiplicities of infection of defective interfering particles, the inhibition of host cell protein synthesis by VSV decreased by a maximum of only 10%. The data also indicated that under these conditions the protein-synthesizing capacity of the cells was not exhausted. We concluded that competition for cellular ribosomes could not have been the major factor in the inhibition of host cell protein synthesis by VSV. This conclusion was further supported by inhibition data obtained with VSV mutants. The ts G22 mutant, defective in replication but not in primary transcription, inhibited host protein synthesis at the nonpermissive temperature (39 degrees C) to the same extent as did wild-type virus, even though it generated only 30 to 50% of the amount of viral mRNA as did wild-type virus. Conversely, in infections with the R1 mutant, which did not inhibit host cell protein synthesis, the amount of total and polysome-bound viral mRNA was indistinguishable from that obtained in infections by wild-type virus. PMID- 6296432 TI - Comparison of the oligosaccharide moieties of the major envelope glycoproteins of the subgroup A and subgroup B avian myeloblastosis-associated viruses. AB - The nature of the oligosaccharide chains of the major envelope glycoprotein, gp85, from avian myeloblastosis-associated viruses has been examined for the subgroup A and subgroup B viruses replicated in fibroblasts from the same chicken embryos. Pronase-digested glycopeptides from [3H]mannose- or [3H]glucosamine labeled viruses were analyzed by the combined techniques of gel filtration, endo beta-N-acetylglucosaminidase digestion, and concanavalin A affinity chromatography. The gp85 protein from these two viruses, and also from another subgroup A avian leukosis virus replicated in the same cells, contained a diverse array of asparagine-linked oligosaccharides of the acidic type [(sialic acid +/- galactose-N-acetylglucosamine)2-4-(mannose)3-N-acetylglucosamine2(+/- fucose) asparagine], hybrid type (sialic acid +/- galactose-N-acetylglucosamine (mannose)5,4-N-acetylglucosamine2-asparagine), and neutral type [(mannose)5-9-N acetylglucosamine2-asparagine], with the more highly branched (tri or tetraantennary or both) acidic-type structures representing the predominant class of oligosaccharide. Minor differences were observed between the gp85 of the subgroup B versus subgroup A viruses. PMID- 6296433 TI - Herpes simplex virus types 1 and 2 induce shutoff of host protein synthesis by different mechanisms in Friend erythroleukemia cells. AB - Herpes simplex virus type 1 (HSV-1) and HSV-2 disrupt host protein synthesis after viral infection. We have treated both viral types with agents which prevent transcription of the viral genome and used these treated viruses to infect induced Friend erythroleukemia cells. By measuring the changes in globin synthesis after infection, we have determined whether expression of the viral genome precedes the shutoff of host protein synthesis or whether the inhibitor molecule enters the cells as part of the virion. HSV-2-induced shutoff of host protein synthesis was insensitive to the effects of shortwave (254-nm) UV light and actinomycin D. Both of the treatments inhibited HSV-1-induced host protein shutoff. Likewise, treatment of HSV-1 with the cross-linking agent 4,5',8 trimethylpsoralen and longwave (360-nm) UV light prevented HSV-1 from inhibiting cellular protein synthesis. Treatment of HSV-2 with 4,5',8-trimethylpsoralen did not affect the ability of the virus to interfere with host protein synthesis, except at the highest doses of longwave UV light. It was determined that the highest longwave UV dosage damaged the HSV-2 virion as well as cross-linking the viral DNA. The results suggest that HSV-2 uses a virion-associated component to inhibit host protein synthesis and that HSV-1 requires the expression of the viral genome to cause cellular protein synthesis shutoff. PMID- 6296434 TI - Polypeptide structure and encoding location of the adenovirus serotype 2 late, nonstructural 33K protein. AB - Radiochemical microsequence analysis of selected tryptic peptides of the adenovirus type 2 33K nonstructural protein has revealed the precise region of the genomic nucleotide sequence that encodes this protein. The initiation codon for the 33K protein lies 606 nucleotides to the right of the EcoRI restriction site at 70.7 map units and 281 nucleotides to the left of the postulated carboxyterminal codon of the adenovirus 100K protein. The coding regions for these two proteins thus overlap; however, the 33K protein is derived from the +1 frame with respect to the postulated 100K reading frame. Our results contradict an earlier published report suggesting that these two proteins share extensive amino acid sequence homology (N. Axelrod, Virology 87:366-383, 1978). The published nucleotide sequence of the Ad2 EcoRI-F fragment (70.7 to 75.9 map units) cannot accommodate in a single reading frame the peptide sequences of the 33K protein that we have determined. Sequence analysis of DNA fragments derived from virus has confirmed the published nucleotide sequence in all critical regions with respect to the coding region for the 33K protein. Consequently, our data are only consistent with the existence of an mRNA splice within the coding region for 33K. Consensus donor and acceptor splice sequences have been located that would predict the removal of 202 nucleotides from the transcripts for the 33K protein. Removal of these nucleotides would explain the structure of a peptide that cannot otherwise be directly encoded by the EcoRI-F fragment. Identification of the precise splice points by peptide sequencing has permitted a prediction of the complete amino acid sequence for the 33K protein. PMID- 6296435 TI - Characterization of the 5' termini of purified nascent simian virus 40 late transcripts. AB - The primary transcripts of simian virus 40 are extensively processed in the nuclei of infected monkey cells before they are transported to the cytoplasm as mature mRNAs. To investigate the early steps in this process, in particular, to determine which events occur on nascent chains before the termination of transcription, we have developed a procedure for the purification of nascent viral transcripts. This technique involves the in vitro incorporation of mercurated residues into the growing 3' ends of pre-initiated nascent chains, allowing their specific purification by sulfhydrylcellulose affinity chromatography. We further selected viral specific transcripts by hybridization to simian virus 40 DNA-cellulose. We describe here our analysis of the 5' termini of purified nascent simian virus 40 transcripts. This analysis revealed various cap structures, providing direct evidence that primary viral transcripts are capped before chain completion. The various cap structures exhibited a full range of methylation states. Completely unmethylated GpppA cap cores were identified, as well as caps methylated at the penultimate position only. The presence of GpppAm and GpppmAm caps indicates that, in BSC-1 cells, the penultimate nucleotide can be methylated before 7-methyl-G formation. Furthermore, the proportions of the various intermediates suggest that, in contrast to the viral capping enzymes of vaccinia virus and reovirus, the cellular enzymes methylate in the following order: GpppA leads to GpppAm leads to GpppmAm leads to 7mGpppmAm. In addition to capped ends, we also detected some unprocessed pppA ends. To our knowledge, this is the first time uncapped termini have been identified on RNAs known to be polymerase II products. PMID- 6296436 TI - Large RNase T1-resistant oligonucleotides encoding p15E and the U3 region of the long terminal repeat distinguish two biological classes of mink cell focus forming type C viruses of inbred mice. AB - We used T1 oligonucleotide maps, in conjunction with available nucleotide sequences of appropriate C-type viruses, to identify regions of the viral genome that distinguish two biological classes of mink cell focus-forming (MCF) viruses described previously by Cloyd et al. (J. Exp. Med. 151:542-522, 1980). We found that leukemogenic MCF viruses from thymus differed from non-leukemogenic MCFs isolated from nonthymic neoplasms in nucleotide sequences encoding Prp15E and the U3 portion of the long terminal repeat (LTR). The thymic isolates possessed recombinant Prp15E genes, with the 5' to mid portion derived from their ecotropic parents and the extreme 3' portion invariably derived from their nonecotropic parents. These viruses probably derived the entire U3 portion of their LTRs from their nonecotropic parents. The nonthymic MCFs appeared to inherit their entire Prp15E coding region from their nonecotropic parents. We failed to detect consistent differences in gp70-coding sequences between the two groups of MCFs, but this may simply reflect limitations of the data. The studies presented here, in conjunction with studies from a number of labs indicating a role for MCF gp70 in leukemogenesis, indicate that three genetic elements, gp70, p15E, and the U3 portion of the LTR, may all play a role in determining the leukemogenic phenotype of type C viruses of high-leukemic inbred mice. PMID- 6296437 TI - Nucleotide sequence of the 3' end of MCF 247 murine leukemia virus. AB - We isolated DNA clones of MCF 247, a leukemogenic, recombinant type C virus obtained from the thymus of an AKR mouse. We determined the nucleotide sequence of the viral long terminal repeat (LTR) and the 3' end of env, and we compared the sequences to corresponding sequences of the genome of Akv virus, the putative ecotropic parent of MCF 247. By analogy with Moloney leukemia virus, we identified the amino terminus of Prp15E, the C-terminal proteolytic cleavage product of env and precursor to mature virion p15E. In MCF 247 the presumptive Prp15E is encoded by a 603-nucleotide open reading frame. The majority of this sequence is identical to that of Akv. However, a recombination event near the 3' end of the Prp15E-coding region introduces nonecotropic sequences into MCF 247, and these extend to the 3' end through the U3 portion of the LTR. The U3 regions of Akv and MCF 247 are about 83% homologous. The R and U5 regions of the LTR of MCF 247 and Akv are identical. Large RNase T1-resistant oligonucleotides analyzed previously in numerous ecotropic and MCF viral genomes were located within the Akv and MCF 247 DNA sequences. The resulting precise T1 oligonucleotide maps of the 3' ends of MCF viral genomes reveal that the biologically defined, leukemogenic class I MCFs isolated from thymic neoplasms of inbred mice all share the sequence pattern seen in MCF 247, a representative of this group; they possess recombinant Prp15E genes and derive U3 from their nonecotropic parents. PMID- 6296439 TI - Improved localization of phosphorylation sites in simian virus 40 large T antigen. AB - The location of phosphorylation sites in the large T antigen of simian virus 40 has been studied both by partial chemical cleavage and by partial proteolysis of various forms of large T. These included the full-size wild-type molecule with an apparent molecular weight of 88,000, deleted molecules coded for by the mutants dl1265 and dl1263, and several shortened derivatives generated by the action of a cellular protease. These molecules differed from each other by variations in the carboxy-terminal end. In contrast, a ubiquitous but minor large T form with a molecular weight of 91,000 was found to be modified in the amino-terminal half of the molecule. In addition to the phosphorylation of threonine at position 701 (K. H. Scheidtmann et al., J. Virol. 38:59-69, 1981), two other discrete domains of phosphorylation were recognized, one at either side of the molecule. The amino terminal region was located between positions 81 and 124 and contained both phosphothreonine and phosphoserine residues. The carboxy-terminal region was located between approximate positions 500 and 640 and contained at least one phosphoserine residue but no phosphothreonine. The presence in the phosphorylated domains of large T of known recognition sequences for different types of protein kinases is discussed, together with possible functions of large T associated with these domains. PMID- 6296438 TI - Comparison of infectious JC virus DNAs cloned from human brain. AB - We cloned JC virus DNA obtained directly from brain tissue of 10 cases of progressive multifocal leukoencephalopathy and compared DNAs by restriction endonuclease mapping. Before cloning, each DNA preparation was homogeneous with respect to restriction patterns, but with the cloned DNAs we found variability in three regions of the genome among DNAs from different cases. There was a region of hypervariability between 0.67 and 0.725 map units; no two DNAs were exactly alike in this region. We determined that the origin of DNA replication also was in this region at 0.69 +/- 0.02 map units. In 4 of the 10 DNAs examined there was a deletion of approximately 75 base pairs between 0.14 and 0.235 map units, the region presumed to contain the codons for the C-terminal ends of the structural protein Vpl and for T antigen. JC virus DNA from these same four cases had an additional HincII-HpaI site at 0.895 map units in the presumptive Vp3 and Vp2 coding regions. Overall, no two JC virus genomes were identical although all were from fatal central nervous system infections and were infectious in vitro. Our restriction patterns suggest that there are two subtypes of JC virus circulating in the population. PMID- 6296440 TI - Characterization of herpes simplex virus 2 temperature-sensitive mutants whose lesions map in or near the coding sequences for the major DNA-binding protein. AB - By marker rescue with cloned herpes simplex virus 2 DNA fragments, we have mapped the temperature-sensitive mutations of a series of herpes simplex virus 2 mutants to a region of the herpes simplex virus 2 genome that lies within or near the coding sequences for the major DNA-binding protein, ICP8. In cells infected with certain of these mutants at the nonpermissive temperature, the association of the major DNA-binding protein with the cell nucleus was defective. In these cells, the DNA-binding protein accumulated in the cytoplasmic and the crude nuclear detergent wash fractions. At the permissive temperature, the maturation of the mutant ICP8 was similar to that of the wild-type viral protein. With the remainder of the mutants, the nuclear maturation of ICP8 was similar to that encoded by the wild-type virus at the nonpermissive and permissive temperatures as assayed by cell fractionation. PMID- 6296441 TI - Genetic analysis of temperature-sensitive mutants which define the genes for the major herpes simplex virus type 2 DNA-binding protein and a new late function. AB - Eleven temperature-sensitive mutants of herpes simplex virus type 2 (HSV-2) exhibit overlapping patterns of complementation that define four functional groups. Recombination tests confirmed the assignment of mutants to complementation groups 1 through 4 and permitted the four groups to be ordered in an unambiguous linear array. Combined recombination and marker rescue tests (A. E. Spang, P. J. Godowski, and D. M. Knipe, J. Virol. 45:332-342, 1983) indicate that the mutations lie in a tight cluster near the center of UL to the left of the gene for DNA polymerase in the order 4-3-2-1-polymerase. The seven mutants that make up groups 1 and 2 fail to complement each other and mutants in HSV-1 complementation group 1-1, the group thought to define the structural gene for the major HSV-1 DNA-binding protein with a molecular weight of 130,000. At 38 degrees C, mutants in groups 1 and 2 synthesize little or no viral DNA, and unlike cells infected with the wild-type virus, mutant-infected cells exhibit no detectable nuclear antigen reactive with monoclonal or polypeptide-specific antibody to the major HSV-2 DNA-binding protein. The four mutants that make up groups 3 and 4 do not complement each other, nor do they complement mutants in group 2. They do, however, complement mutants in group 1 as well as representative mutants of HSV-1 complementation group 1-1. At 38 degrees C, mutants in groups 3 and 4 are phenotypically DNA+, and nuclei of mutant-infected cells contain the HSV-2 DNA-binding protein. Thus, the four functional groups appear to define two closely linked genes, one encoding an early viral function affecting both viral DNA synthesis and expression of the DNA-binding protein with a molecular weight of 130,000 (groups 1 and 2), and the other encoding a previously unidentified late viral function (groups 3 and 4). The former gene presumably represents the structural gene for the major HSV-2 DNA-binding protein. PMID- 6296442 TI - Genetic analysis of temperature-sensitive mutants which define the gene for the major herpes simplex virus type 1 DNA-binding protein. AB - We have assigned eight temperature-sensitive mutants of herpes simplex virus type 1 to complementation group 1-1. Members of this group fail to complement mutants in herpes simplex virus type 2 complementation group 2-2. The mutation of one member of group 1-1, tsHA1 of strain mP, has been shown to map in or near the sequence which encodes the major herpes simplex virus type 1 DNA-binding protein (Conley et al., J. Virol. 37:191-206, 1981). The mutations of five other members of group 1-1 map in or near the sequence in which the tsHA1 mutation maps, a sequence which lies near the center of UL between the genes for the viral DNA polymerase and viral glycoprotein gAgB. These mutants can be divided into two groups; the mutations of one group map between coordinates 0.385 and 0.398, and the mutations of the other group map between coordinates 0.398 and 0.413. At the nonpermissive temperature mutants in group 1-1 are viral DNA negative, and mutant infected cells fail to react with monoclonal antibody to the 130,000-dalton DNA binding protein. Taken together, these data indicate that mutants in complementation groups 1-1 and 2-2 define the gene for the major herpes simplex virus DNA-binding protein, an early gene product required for viral DNA synthesis. PMID- 6296443 TI - Mutants deleted in the agnogene of simian virus 40 define a new complementation group. AB - Analysis of the DNA sequence of the late leader region of simian virus 40 indicates that it might encode a 61-amino acid, highly basic protein, LP-1. Mutants deleted in this region are viable, but they produce infectious progeny more slowly than wild-type virus in established monkey cells. On the basis of the rates of appearance and the sizes of mixed plaques formed after cotransfections with pairs of mutants, we found that mutants defective in the synthesis of LP-1 complementation was also observed in infections with virions and was bidirectional. Therefore, these mutants define a new complementation group, group G. In addition, a protein of the appropriate molecular weight for LP-1 (approximately 8 X 10(3) ) was synthesized by wild-type virus-infected cells but not by mock-infected or group G gene mutant-infected cells. This protein, whose identity has been established definitively by Jay et al. (Nature (London) 291:346 349, 1981), was synthesized at a high rate at late times after infection, was present predominantly in the cytoplasmic fraction of cells, possessed a fairly short half-life, and was absent from mature virions. Once formed, virions of group G gene mutants behaved biologically and physically like virions of wild type virus. On the basis of these findings and other known properties of LP-1 and mutants defective in LP-1 synthesis, we hypothesize that LP-1 functions to facilitate virion assembly, possibly by serving as a nonreusable scaffolding protein. PMID- 6296444 TI - DNA-mediated gene transfer in Friend leukemia cells by cotransfection of simian virus 40 DNA with herpes simplex virus thymidine kinase DNA. AB - Thymidine kinase-negative Friend leukemia cells were cotransfected with simian virus 40 (SV40) DNA and thymidine kinase gene DNA of herpes simplex virus type 1. The transfected thymidine kinase-positive cells were selected in HAT medium, and SV40 T-antigen expression was observed over many months in cells cultured under selective conditions, and after adaptation to normal growth medium under nonselective conditions. It was shown by Southern blot hybridization that SV40 DNA was integrated in multiple copies in the chromosomal DNA of several clones. All SV40 DNA-containing Friend leukemia cell clones analyzed were able to undergo induced erythroid differentiation. Induced cultures still expressed SV40 T antigen to the same extent that untreated control cultures did. PMID- 6296445 TI - Molecular genetics of herpes simplex virus. VIII. further characterization of a temperature-sensitive mutant defective in release of viral DNA and in other stages of the viral reproductive cycle. AB - Previous studies have shown that cells infected with the herpes simplex virus 1(HFEM) mutant tsB7 and maintained at the nonpermissive temperature fail to accumulate viral polypeptides. Analyses of intertypic recombinants generated by marker rescue of tsB7 with herpes simplex virus 2 DNA fragments localized the mutation between 0.46 and 0.52 map units on the viral genome (Knipe et al., J. Virol. 38:539-547, 1981). In this paper we report that the mutation in tsB7 affects several aspects of the reproductive cycle of the virus at the nonpermissive temperature. Thus, (i) viral capsids accumulate at the nuclear pores and do not release viral DNA for at least 6 h postinfection at 39 degrees C. The DNA was released within 30 min after a shift to the permissive temperature. (ii) Experiments involving shifts from the permissive to the nonpermissive temperature indicated that viral protein synthesis was not sustained in cells maintained at the permissive temperature for less than 4 h. (iii) Viral DNA synthesis was delayed at the permissive temperature for as long as 8 h. Once initiated, it continued at 39 degrees C. (iv) Marker rescue of tsB7 by transfection with herpes simplex virus 1(F) DNA fragments localized the mutation to between 0.501 and 0.503 map units on the viral genome. These results are consistent with the tsB7 lesion being in a gene coding for a virion component which affects release of viral DNA from capsids and onset of viral DNA synthesis. PMID- 6296446 TI - Expression of adenovirus type 12 E1A gene in monkey cells, using a simian virus 40 vector. AB - Simian virus 40 (SV40) recombinants carrying the adenovirus type 12 E1A gene were constructed. The SV40 expression vector was constructed by removing most of the VP1 gene and an internal part of the intervening sequence for late 16S RNA and by joining the 5' and 3' splice sites into a small segment. The adenovirus type 12 E1A gene with or without its own promoter was inserted downstream from the SV40 late promoter and the splicing junctions. The recombinant DNA was propagated and packaged in monkey cells by cotransfection with an early temperature-sensitive mutant (tsA58) DNA as helper. Immunofluorescent staining of the monkey cells infected with the resulting virus stocks showed that up to 20% of the cells overproduced the E1A gene products in the nuclei. Two-dimensional gel electrophoresis of the products indicated that the products were very similar or identical to the authentic polypeptides synthesized in adenovirus type 12 infected human embryo kidney cells. The E1A mRNA was initiated at the SV40 late promoter irrespective of the presence of the E1A promoter and terminated at either the E1A or the SV40 polyadenylation signal. These hybrid mRNAs were correctly spliced in the E1A coding region. PMID- 6296447 TI - Hyperproduction of adenovirus type 12 E1B gene product in monkey cells, using a simian virus 40 vector. AB - Simian virus 40 recombinant DNAs carrying the adenovirus type 12 E1B gene were constructed, propagated, and packaged in monkey cells. Monkey cells infected with the resulting virus stocks hyperproduced the E1B gene products in more than 80% of the cells as revealed by immunofluorescence. The products were distributed in both the nuclei and the cytoplasm, and a condensed form of fleck structure was observed in the cytoplasm. Polyacrylamide gel electrophoresis of the cell extracts and their immunoprecipitates detected the E1B-coded 19,000-molecular weight protein but not the 50,000-molecular-weight protein. The 19,000-molecular weight protein and the simian virus 40 VP1 protein were synthesized in nearly equal amounts. PMID- 6296448 TI - Subcellular localization of the simian virus 40 agnoprotein. AB - The intracellular distribution of the simian virus 40 agnoprotein in infected cells was determined by indirect immunofluorescence and biochemical fractionation followed by indirect immunoprecipitation. The specific antibodies used in these studies were directed against either purified agnoprotein or a synthetic oligopeptide homologous to the N-terminus of the processed protein. Both procedures showed predominant localization of the agnoprotein to the cytosol and to the perinuclear region in association with the outer nuclear membrane. A minor but detectable fraction of the protein was also found in the nucleus. The definition of its subcellular distribution, as well as its high lability in vivo and affinity for nucleic acid, provide a basis for speculation on the function of this gene product. PMID- 6296449 TI - The influenza virus nucleoprotein synthesized from cloned DNA in a simian virus 40 vector is detected in the nucleus. AB - We obtained DNA sequences coding for the nucleoprotein (NP) of an influenza A virus by reverse transcription of virion RNA with synthetic oligonucleotide primers. Terminal sequence analysis showed that the cloned gene contained a full length copy of the virion RNA segment. The NP-specific DNA was inserted into the late region of a simian virus 40 vector, and the DNA recombinant was propagated in the presence of an early simian virus 40 temperature-sensitive mutant helper. Infection of African green monkey kidney cells with the recombinant produced a polypeptide immunoprecipitable with NP-specific antisera. The polypeptide product had a molecular weight of 56,000, identical to that of the nucleoprotein of influenza virus as estimated on polyacrylamide gels. The putative NP was detected in the nucleus of infected primate cells by an immunofluorescence assay. This nuclear localization of NP from recombinant DNA was similar to that seen during influenza virus infection. PMID- 6296450 TI - Processing of the encephalomyocarditis virus capsid precursor protein studied in rabbit reticulocyte lysates incubated with N-formyl-[35S]methionine-tRNAfMet. AB - Translation of encephalomyocarditis virus RNA in rabbit reticulocyte lysates in the presence of N-formyl-[(35)S]methionine-tRNA(f) (Met) revealed that a small polypeptide is cleaved from the N-terminus of the capsid protein precursor, preA, by virus-coded protease activity. Therefore, this N-terminal segment comprising the translation initiation site is not conserved in any of the mature capsid proteins. PMID- 6296451 TI - Metabolic turnover of phosphorylation sites in simian virus 40 large T antigen. AB - Four (groups of) phosphorylation sites exist in the large T antigen of simian virus 40, and they involve at least two serine and two threonine residues (Van Roy et al. J. Virol. 45:315-331, 1983). All the phosphorylation sites were found to be modified and again dephosphorylated at discrete rates, with phosphoserine residues having the highest turnover rate. The measured half-lives ranged between 3 h (for the carboxy-terminal phosphoserine site) and 5.5 h (for the amino terminal phosphothreonine site). The influence of four temperature-sensitive A mutations on phosphorylation of large T antigen was also examined. At restrictive temperature, phosphorylation of the carboxy-terminal phosphoserine in mutated large T antigen was found to be particularly impaired. These data emphasize the physiological importance of the latter phosphorylation site. PMID- 6296452 TI - Isolation and characterization of mink cell focus-inducing murine leukemia viruses with xenotropic host range from mouse strain SL. AB - A new type of mink cell focus-inducing virus was persistently isolated from the leukemic tissues of SL mice. In contrast to the dual tropic mink cell focus inducing viruses reported to date, the new virus has the host range of the xenotropic murine leukemia virus. Analysis of RNase T(1) fingerprints of genomic RNAs suggested that the mink cell focus-inducing virus with the xenotropic host range isolated from SL mice is a recombinant virus deriving from xenotropic murine leukemia virus. PMID- 6296453 TI - Nucleotide sequences of feline sarcoma virus long terminal repeats and 5' leaders show extensive homology to those of other mammalian retroviruses. AB - The nucleotide sequences of the Gardner-Arnstein feline sarcoma virus (FeSV) long terminal repeat and the adjacent leader sequences 5' to the viral gag gene were determined. These were compared with homologous portions of Synder-Theilen FeSV and with previously published sequences for Moloney murine sarcoma virus and simian sarcoma virus proviral DNA. More than 75% of the residues in the FeSV R and U5 regions were homologous to sequences within the same regions of the other viral long terminal repeats. Unexpectedly, alignment of the FeSV sequences with those of the Moloney murine sarcoma and simian sarcoma viruses showed similar extents of homology within U3. The homologous U3 regions included the inverted repeats, a single set of putative enhancer sequences, corresponding to a "72-base pair" repeat, and sequences, including the CAT and TATA boxes, characteristic of eucaryotic promotors. The 5' leader sequences of both FeSV strains included a binding site for prolyl tRNA and a putative splice donor sequence. In addition, the FeSV leader contained a long open reading frame which was adjacent to and in phase with the ATG codon at the 5' end of the FeSV gag gene. The open reading frame could code for a signal peptide of about 7.4 kilodaltons. Our results support the concept that the virogenic portions of both FeSV and simian sarcoma virus were ancestrally derived from viruses of rodent origin, with conservation of regulatory sequences as well as the viral structural genes. PMID- 6296455 TI - Organization and stability of endogenous xenotropic murine leukemia virus proviral DNA in mouse genomes. AB - In a series of blot hybridization experiments, using a xenotropic envelope probe and restriction enzymes known to cut xenotropic proviral DNA a single time (EcoRI) or not at all (HindIII), we have studied the organization and relationship of endogenous xenotropic env-related sequences in various mouse strains. Multiple copies (18 to 28) of xenotropic env-reactive fragments were found in all mouse DNAs after digestion with either HindIII or EcoRI, and the majority of fragments were of sizes compatible with their origin from full-length proviral DNA. Five HindIII and five EcoRI restriction fragments were common to all inbred mouse DNAs tested. In addition, each strain exhibited unique characteristic xenotropic env-reactive bands; these bands were remarkably stable during many years of inbreeding. The cleavage patterns characteristic of each strain were also useful for showing genealogical relatedness among the various inbred mice. PMID- 6296454 TI - Control of RFM strain endogenous retrovirus in RFM mouse cells. AB - RFM/Un mice express an endogenous type C retrovirus throughout their life span in many tissues; primary or established embryo fibroblast cell cultures do not express a virus but can be induced by exposure to 5-iodo-2'-deoxyuridine. All of our sources yielded a single ecotropic virus (RFV) which appeared to be related more closely to the endogenous N-tropic virus (WN1802N) of BALB/c mice than to Gross leukemia virus on the basis of two-dimensional gel electropherograms of virion proteins. No xenotropic or recombinant viruses were isolated by cocultivation techniques. RFV is N-tropic, and RFM/Un cells possess the Fv-1n allele, as indicated by restriction of B-tropic virus and susceptibility to Gross strain N-tropic virus. However, RFM cells are highly resistant to RFV and other endogenous N-tropic viruses. This resistance is expressed by two-hit titration kinetics and by inhibition of viral linear duplex DNA formation. This is similar to the effects of the Fv-1 locus, but preliminary work has shown no apparent genetic linkage between the two restrictions. The relative strength of the restriction, the presence of a single class of ecotropic virus, and the absence of recombinant viruses suggest that in RFM mice virus is expressed only in cells in which it is induced and not by cell-to-cell transmission. PMID- 6296456 TI - Simian virus 40 promoters direct expression of the tetracycline gene in plasmid pACYC184. AB - Insertion of HindIII DNA fragments into the HindIII site of plasmid pACYC184 destroys the promoter of the plasmid tetracycline resistance gene and causes Escherichia coli cells harboring recombinant plasmids to be tetracycline sensitive and chloramphenicol resistant. The HindIII-C DNA fragment of simian virus 40 contains the two virus promoters and the virus origin of replication. We report the isolation of recombinant plasmids that contained the simian virus 40 HindIII-C DNA fragment at the HindIII site but were capable of conferring tetracycline resistance to E. coli cells. The viral promoter sequences contained in the HindIII-C fragment presumably replaced the inactivated tetracycline resistance gene promoter sequences and enabled transcription of the tetracycline resistance gene. PMID- 6296457 TI - Epstein-Barr HR-1 virion DNA is very highly methylated. AB - The virion DNA of Epstein-Barr virus strain HR-1 was found to be methylated to a very large extent, approaching 15%. This extent of methylation is in great contrast to other classes of tumor viruses, and it is the highest that we know of in any mammalian system. PMID- 6296458 TI - Analysis of recombinant DNA clones of the endogenous BALB/c murine leukemia virus WN1802N: variation in long terminal repeat length. AB - We analyzed 15 recombinant DNA clones of the unintegrated closed circular DNA intermediate of the BALB/c endogenous ecotropic murine leukemia virus WN1802N. Thirteen of these clones had an insert which corresponded to the complete murine leukemia virus genome. Of these, six contained a single long terminal repeat (LTR) and seven contained two LTRs. The viral genomes in nine clones had an LTR of 520 base pairs (bp), one had an LTR of 570 bp, three had an LTR of 600 bp, and one had an LTR of 670 bp. Restriction endonuclease analysis demonstrated that the size variability resides in the U3 region. Seven of eight clones which yielded infectious virus by DNA transfection had the 520-bp LTR, and the other had a 600 bp LTR. More detailed examination of plasmid subclones of three isolates with different-sized LTRs revealed that the approximate position which varies in the U3 region corresponds to the 72-bp repeat region of Moloney sarcoma virus. Possible consequences of these variations are discussed. PMID- 6296459 TI - Vaccinia virus thymidine kinase and neighboring genes: mRNAs and polypeptides of wild-type virus and putative nonsense mutants. AB - Enzymatically active thymidine kinase (TK) was made in reticulocyte lysates programmed with early vaccinia mRNA that hybridized to plasmid recombinants containing either of two adjacent small DNA subsegments of the viral HindIII-J fragment. The map position of an early polypeptide, with a molecular weight of 19,000 (19K), coincided precisely with that of the TK. The absence of the 19K polypeptide in cell-free translation products of hybridization-selected mRNAs from several TK-negative mutants provided an independent identification of the TK polypeptide. The small size of the TK polypeptide of vaccinia virus distinguishes it from that of procaryotes, eucaryotes, and herpesvirus. Five early mRNAs of 3,840, 2,390, 1,790, 1,070, and 590 nucleotides were mapped within the HindIII-J fragment by RNA blotting and nuclease S1 digestion of RNA-DNA hybrids. The RNAs of 590 and 2,380 nucleotides were found to have 5' coterminal ends and represent major and minor forms, respectively, of the TK message. The 3' end of the minor TK mRNA appeared to be coterminal with the 3' end of the 1,790-nucleotide transcript which encodes a 41K polypeptide. The 1,070-nucleotide RNA was identified as the message for a 21K polypeptide. All of these RNAs, including the two forms of the TK message, were made by the putative TK-negative nonsense mutants. PMID- 6296460 TI - Genomic structure of human polyoma virus JC: nucleotide sequence of the region containing replication origin and small-T-antigen gene. AB - The nucleotide sequence of the region of human polyoma virus JC DNA between 0.5 and 0.7 map units from a unique EcoRI cleavage site was determined and compared with those of the corresponding regions of another human polyoma virus, BK, and simian virus 40 DNAs. Within this region consisting of 945 base pairs, we located the origin of DNA replication near 0.7 map units, the entire coding region for small T antigen, and the splice junctions for large-T-antigen mRNA. The deduced amino acid sequences for small T antigen and the part of large T antigen markedly resembled those of polyoma virus BK and simian virus 40. The results strongly suggest that polyoma virus JC has the same organization of early genome as polyoma virus BK and simian virus 40 on the physical map, with the EcoRI site as a reference point. PMID- 6296461 TI - Characterization of the soluble glycoprotein released from vesicular stomatitis virus-infected cells. AB - Vesicular stomatitis virus-infected Chinese hamster ovary cells release into the extracellular medium a soluble form of the vesicular stomatitis virus glycoprotein (G protein) termed Gs (Kang and Prevec, Virology 46:678-680, 1971). The properties of this molecule and the cellular site at which it is generated were characterized. By comparing the sizes and the peptide maps of the unglycosylated forms of G and Gs, we found that between 5,000 and 6,000 daltons of the carboxy-terminal end of the G protein is cleaved to generate the Gs molecule. This truncated molecule contains no fatty acid. Gs released from cells grown at 39 degrees C migrated on polyacrylamide gels slightly slower than Gs released at 30 degrees C. The unglycosylated form of Gs also showed this size difference. Furthermore, unglycosylated Gs was resolved into two species upon isoelectric focusing: the relative amounts of the two species depended upon the temperature at which infected cells were incubated. Full-sized unglycosylated virus-associated G also was resolved into two species, but the more basic form predominated at both 30 and 39 degrees C. The appearance of Gs in the extracellular medium depended upon the presence of stable, full-sized G at the cell surface. The amount of Gs released was quantitated in seven different situations in which the migration of G to the cell surface was inhibited. In all cases, the amount of Gs released was also decreased. In addition, incubation of cells surface labeled with 125I resulted in the release of 125I-labeled Gs protein, as well as full-sized G protein. These results suggest that Gs is generated primarily by proteolytic cleavage of plasma membrane-associated G at a site in the molecule just amino terminal to the membrane-spanning region of the molecule. PMID- 6296462 TI - Palindromic adenovirus type 5-simian virus 40 hybrid. AB - A family of novel adenovirus type 5-simian virus 40 (Ad5-SV40) recombinants (Ad5++D1) whose genomes consist of symmetrically inverted structures was isolated. Particles of Ad5++D1 could contain one of several recombinant genomes that differed incrementally from one another by a full-length copy of linear SV40 DNA. The members of the Ad5++D1 family appeared to be in genetic equilibrium with one another. In all probability this equilibrium was maintained by homologous recombination, resulting in the loss or gain of one or two unit length copies of the SV40 genome. The genome of the most abundant recombinant from consisted of a giant inverted repeat which was some 35,000 nucleotide pairs in length. Beginning from one end, the recombinant genome consisted of 3,534 nucleotides derived from the left end of the adenovirus type 5 genome; these nucleotides were joined to 2.7 copies of SV40 DNA arranged as head-to-tail tandems. This entire structure was then repeated in the opposite orientation, thereby forming a large inverted repeat whose structure was Ad5-SV40-SV40-04VS-04VS-5dA. The population of hybrid genomes was stable and was maintained through serial rounds of infection. PMID- 6296463 TI - The treatment of stages I and II nonseminomatous testicular tumors: the Buffalo experience from 1970 to 1979. AB - We reviewed our experience in the treatment of stages I and II nonseminomatous testicular tumors to evaluate new trends and improvements in this disease. Of the patients with stage I disease there was a significantly higher percentage of survivors among those who received adjunctive chemotherapy with actinomycin D. Since these patients were not randomized prospectively this observation should be taken only as a suggestion of the possible effectiveness of adjunctive chemotherapy in this group. However, the greatest advances have been observed among patients with stage IIB disease. Since the establishment of modern accepted combination chemotherapy survival free of disease (minimal followup 2 years) in this group has tripled. PMID- 6296464 TI - Control of recurrent lower urinary tract infection in the postmenopausal woman. AB - There were 5 postmenopausal women with severe recurrent cystitis who were placed on a protocol of intravaginal estrogen in an effort to control infection. Before therapy all patients had colonization of the vagina by 1 or more species of Enterobacteriaceae and an associated elevation of vaginal pH greater than 5.2 (normal range 4.0 to 4.5). After acute antibiotic treatment and continuous low dose intravaginal estrogen all 5 patients showed a return of vaginal pH to the normal range and disappearance of vaginal bacterial pathogens. Of the 5 women 4 have had no further urinary tract infection off antibiotics and on low dose intravaginal estrogen. One patient had a urinary tract infection in association with a flare in colitis, which was controlled with reinstitution of antibiotics and high dose intravaginal estrogen therapy. PMID- 6296465 TI - Ceftizoxime in the treatment of urinary tract infections. AB - Ceftizoxime, a new beta-lactamase-resistant, semisynthetic antibiotic, was compared to cefamandole in a prospective randomized trial to determine its efficacy and safety in 21 patients with acute, complicated urinary tract infections. Four patients randomized initially to receive cefamandole were found to have resistant organisms and were treated with ceftizoxime. Dosage for ceftizoxime was 1 gm. administered parenterally every 12 hours, while 1 gm. cefamandole was given every 6 hours. Urine cultures were obtained before the initiation of therapy, on day 4, after completion of therapy and 4 to 6 weeks after therapy. Specified laboratory tests were obtained. Of 14 patients receiving ceftizoxime 11 (79 per cent) and of 7 patients receiving cefamandole 7 (100 per cent) had negative cultures at the completion of therapy and 4 to 6 weeks later. No patient had any adverse reaction to ceftizoxime. Ceftizoxime is a safe and effective antibiotic agent when used as a single agent for complicated urinary tract infections. However, ceftizoxime is much more expensive than cefamandole therapy. Therefore, it is recommended that ceftizoxime be reserved for treatment of urinary tract infections stemming from pathogenic species resistant to the less expensive antimicrobials. PMID- 6296466 TI - Retroperitoneal hemorrhage, gross hematuria and painful right flank mass in a 55 year-old man with cirrhosis. PMID- 6296467 TI - Adult Wilms tumor and bilateral germ cell tumors of testes: a case report. PMID- 6296468 TI - Atropine resistance of transmurally stimulated isolated human bladder muscle. AB - Human detrusor strips were obtained from patients undergoing reimplantation of ureters because of reflux, transvesical prostatectomy, or cysto-urethrectomy en bloc because of bladder malignancy. The strips were electrically stimulated. A frequency-dependent contractant response was obtained that was potentiated by physostigmine and abolished by tetrodotoxin. The maximum response approximately equaled that of acetylcholine in a maximum concentration. In most bladder preparations from patients without known functional bladder disturbances, atropine (0.01 to 0.1 microM) had a marked inhibitory effect, and at concentrations exceeding 1 microM the blockade was complete. In strips obtained from patients undergoing transvesical prostatectomy, and who also had a cystometrically verified unstable bladder, there was a varying degree of atropine resistance, with some preparations showing a 50 per cent resistance to atropine. Prazosin, phentolamine, yohimbine, guanethidine, clonidine, and noradrenaline had no consistent effects on the electrically induced bladder contraction. Nifedipine and nimodipine caused a maximum of 65 per cent inhibition of the response. Addition of nimodipine to atropine-resistant strips when maximum atropine inhibition had been reached abolished the contractions. Omitting calcium from the bath solution rapidly abolished the electrically induced contraction. It is suggested that in the normal human bladder the contraction induced by electrical stimulation is mainly atropine sensitive. However, in the functionally disturbed bladder, part of the bladder contraction is atropine resistant, a finding that may have clinical implications. PMID- 6296469 TI - Serologic survey for canine distemper and infectious canine hepatitis in wolves in Alaska. AB - Sera from 57 wolves (Canis lupus) in three areas of Alaska were evaluated for evidence of previous exposure to infectious canine hepatitis virus (ICHV) and canine distemper virus (CDV). Fifty-four sera (94.7%) were positive for ICHV exposure and four (7%) were positive for CDV exposure. All four CDV-reacting wolves also had titres to ICHV. The relatively common occurrence of ICHV exposure may be due to the greater resistance of ICHV to chemical and physical agents and its transmissibility via the urine of infected animals. The ICHV titres observed could indicate enzootic pathogenic ICHV, or exposure to the mildly pathogenic vaccine strain of CAV-1 through contact with the urine of domestic dogs. If CAV-1 is the original source of exposure, the titres could represent an ICHV-protected wolf population. PMID- 6296470 TI - Experimental infection of European wild swine (Sus scrofa L.) with pseudorabies virus. AB - Experimental infection with pseudorabies virus was carried out by oral exposure of four young wild swine held in contact with two unexposed controls. No disease was observed but virological procedures indicated that the virus was shed in saliva and, in one case, in the nasal discharge, with subsequent infection of the control animals. After slaughter the virus was reisolated from the tonsils but not from lungs and brain. Virus reisolation from the tonsils was obtained in two animals after the throat swabs became negative. Virus neutralizing antibodies were detected. PMID- 6296471 TI - Prevalence of neutralizing antibodies to bovid herpesvirus 2 in African wildlife. AB - A total of 3,470 sera, collected between 1963 and 1980 from 45 different species of wildlife in nine African countries, was examined for virus neutralizing (VN) antibodies to bovid herpesvirus 2. Antibodies were demonstrated in 20 species including 15 Bovidae, two Suidae, hippopotamus (Hippopotamus amphibius), giraffe (Giraffa camelopardalis) and a green monkey (Cercopithecus aethiops); 11 of these species had not been previously recorded as sero-positive. Although the significance of neutralizing antibodies in the absence of virus isolation remains in doubt, results suggest that infection is widespread in wildlife. The highest VN titres were recorded in waterbuck (Kobus ellipsiprymnus and K. defassa), reedbuck (Redunca arundinum) and buffalo (Syncerus caffer). Infection appears to be continuous in free-living populations of buffalo and antibodies are present in the majority of animals by the age of 2 yr. PMID- 6296472 TI - Survival of duck plague virus in water from Lake Andes National Wildlife Refuge, South Dakota. AB - An isolant of duck plague herpesvirus from the Lake Andes Refuge outbreak was seeded in raw and filter-decontaminated water from two locations on the refuge, held at 4 C, and assayed for infectivity intermittently over a period of 2 mo. From an initial level of about 10(5) PFU per ml, infectivity in the filtered samples uniformly dropped to about 10(4) PFU per ml. Infectivity in the raw samples declined much more rapidly; infectious virus remaining at the end of 2 mo (ca. 10(1) PFU per ml) was only about 0.01% of that originally seeded. PMID- 6296474 TI - Spontaneous bacterial peritonitis and pneumoperitoneum. A false surgical emergency. PMID- 6296473 TI - Primary hepatocellular carcinoma in a blacktailed prairie dog (Cynomys ludovicianus). PMID- 6296475 TI - [Antibiotics induced neuromuscular block by chelation of ionized calcium]. PMID- 6296476 TI - [Mechanisms of pain relief following sympathetic ganglion block]. PMID- 6296477 TI - [Clinical use of pivmecillinam in urinary tract infections. 3]. PMID- 6296478 TI - [Clinical studies on cefmenoxime in the intensive care unit]. AB - Two to 6 g of CMX was administered daily to 9 patients who were admitted to ICU, i.e. 5 cases with pneumonia and 4 with sepsis. In all cases, CMX was administered concomitantly with aminoglycoside which had been administered, and additional administration of other antibiotics was avoided. Bacteriologically, P. aeruginosa was isolated from 4 cases, K. pneumoniae from 4 cases, S. marcescens, P. mirabilis and P. cepacia respectively from 1 case. The CMX treatment was considered effective in 4 of 5 pneumonia cases and in 3 of 4 sepsis cases. In total, 7 of 9 cases responded effectively. The clinical effective rate was 77.8%. Elevation of GOT and GPT values was noticed in 1 case, however, the causality with CMX administration was unclear. PMID- 6296479 TI - [Molecular genetics of R plasmid--the role of the host organism in transposition of a transposon, Tn5, and the mechanism of replication control by Inc FII R plasmid]. PMID- 6296480 TI - [Location of exchangeable cations within the spore of Clostridium perfringens]. PMID- 6296481 TI - [Plasma exchange for malignant neoplasms--immunological consideration]. PMID- 6296482 TI - [Cyclic nucleotides and the cardiac function]. PMID- 6296483 TI - [Na+, K+-ATPase and digitalis actions]. PMID- 6296484 TI - [Ontogeny of the renin-angiotensin-aldosterone system]. PMID- 6296485 TI - [Regulatory mechanism of renin secretion]. PMID- 6296486 TI - [Angiotensin converting enzyme]. PMID- 6296487 TI - [Isorenin--renin-angiotensin system of the brain]. PMID- 6296488 TI - [Renin-angiotensin-aldosterone function tests]. PMID- 6296490 TI - [Surgical treatment of primary and metastatic cancer of the liver]. PMID- 6296489 TI - [Major endocrine diseases]. PMID- 6296491 TI - [Angiographic demonstration of the collateral circulation to the liver after arterial embolization of hepatocellular carcinoma]. PMID- 6296492 TI - [Hepatocellular carcinoma with cavernous transformation of the portal vein]. PMID- 6296493 TI - [Radiotherapy of malignant renal tumors]. PMID- 6296494 TI - [Adrenal metastasis of hepatocellular carcinoma]. PMID- 6296495 TI - [2 cases of ectopic gastric mucosa in the upper esophagus with accumulation of 99m Tc04]. PMID- 6296496 TI - [CT and clinical study of intratumoral gas formation following transarterial embolization of hepatoma and renal cell carcinoma]. PMID- 6296497 TI - [Diagnostic pitfalls in angiography of hepatocellular carcinoma]. PMID- 6296498 TI - [Heterotransplantation of human gastric carcinomas into nude mice. III. Histological and histochemical studies on transplantable human signet-ring cell carcinoma in nude mice]. PMID- 6296499 TI - [An autopsy case of WDHA syndrome]. PMID- 6296500 TI - [Isolation of parietal cells using percoll as density gradient medium]. PMID- 6296501 TI - [Electron probe x-ray microanalysis of various effector cells in three types of the human lymphocytic cytotoxicities against Chang liver cells and the human hepatoma cell line--the significance of calcium and magnesium in various effector cells]. PMID- 6296502 TI - [Immunohistochemical study of alpha 1-antitrypsin on malignant hepatoma]. PMID- 6296503 TI - [Intraarterial infusion of cisplatin in liver tumor]. PMID- 6296504 TI - [An experimental study on the neurotoxicity of 2-octanone and 2-hexanol, a metabolite of n-hexane]. AB - An electrophysiological study of the neurotoxicity of 2-octanone (an analogue of methyl n-butyl ketone) and 2-hexanol (a metabolite of n-hexane) was conducted on rats as a part of the study to determine the specific molecular arrangement required for the development of peripheral neuropathy. The compound 2-octanone or 2-hexanol was administered subcutaneously in the daily dose of 400 mg/kg of each compound into the back of seven rats, weighing 290 g, 5 days per week for a period of 21 weeks. Animals treated with 2-octanone for 21 weeks failed to exhibit apparent clinical and neurophysiological evidence except a slight inhibition of weight gain and narcotic effects after treatment with the compound. The same doses of 2-hexanol for 21 weeks caused hypersalivation, gait disturbances, crossing phenomena of hind limbs and a failure of normal growth. Retardation of the conduction velocity in the motor and sensory nerve fibers and the prolonged motor latencies of the tail nerves (distal part) began to appear at the 14th week of the experiment when 9.6 g in the total dose had been given to each animal. These changes were intensified in the subsequent course of the experiment. Our previous experiments and the present results showed that n-hexane barely produced peripheral neuropathy in doses over 10.5 g, and that 2-hexanone (MBK), 2,5-hexanediol or 2,5-hexanedione never failed to produce a neuropathy even in doses less than 9.6 g of each compound. The above results suggest that the neurotoxic potency of 2-hexanol is greater than that of n-hexane but less than that of MBK, 2,5-hexanediol or 2,5-hexanedione. PMID- 6296505 TI - [Pharmacological notes on midwifery. Pregnancy and medication. 5. Organic sulfite compounds used as coronary vasodilators]. PMID- 6296506 TI - [Clinical reliability and limitation of 99m Tc-pyrophosphate myocardial scintigraphy for the assessment of acute myocardial infarction--with special reference to evaluation of the area affected by infarction]. PMID- 6296507 TI - [Estimation of effective liver volume ratios by processing liver scintigraphic data with an on-line computer]. PMID- 6296508 TI - [Test of attenuation correction in the imaging determination of left ventricular ejection fractions by the 1st-pass method]. PMID- 6296509 TI - [Application of a rotating gamma camera to myocardial imaging. 5. Topographic assessment of acute myocardial infarction by 99mTC pyrophosphate myocardial ECT]. PMID- 6296510 TI - [Quantitative evaluation of relative separate renal perfusion and function with 123I-ortho-iodohippurate: A comparison with 99mTc-DMSA and 131I-Hippuran]. PMID- 6296511 TI - [Extra-osseous tumor uptake of bone seeking agents]. PMID- 6296512 TI - Kinetic analysis of the effect of tetraethylammonium on neuromuscular transmission in the frog. PMID- 6296513 TI - Chemical modification of several amino acid residues in the glycine receptor of Onchidium neurons. PMID- 6296514 TI - N-nitroso-N-methylurea-induced mammary tumors in the rat: role of prolactin and a prolactin-lowering drug. AB - Female outbred Sprague-Dawley rats bearing N-nitroso-N-methylurea (NMU)-induced mammary tumors received various endocrine therapies 3 months after the first NMU injection. Rats were divided into 5 groups (15-20 rats/group) and received a 4 week treatment as follows: group 1, controls; group 2, ovariectomized; group 3, 0.5 mg 2-bromoergocryptine (CB-154) injected so twice daily; group 4a, pituitary implant under the kidney capsule; and group 4b, CB-154 injected during the last 2 weeks of the experiment in rats bearing a pituitary implant. Castration of rats with established NMU-induced tumors resulted in a decrease in both tumor number and size, but these parameters again started to increase 3 weeks post castration. CB-154 failed to reduce the tumor number but did arrest the increase in tumor size. In the animals with a pituitary implant, both tumor number and tumor size increased progressively at a greater rate than in control animals, whereas the addition of CB-154 (group 4b) stabilized the tumor growth. Ovariectomy (OVX) resulted in a significant decline of steroid receptor levels. Prolactin (PRL) receptor levels were significantly stimulated by the pituitary implant, and CB 154 prevented this increase. The present studies confirmed that NMU-induced mammary tumors are less hormone-dependent (response to OVX) than 7,12 dimethylbenz[a]anthracene (DMBA)-induced tumors. The role of PRL also appears to be less important, at least for established tumors, for NMU-induced mammary tumors than for DMBA-induced mammary tumors. PMID- 6296515 TI - Differential tumor immunogenicity of DBA/2 mouse lymphoma L1210 and its sublines. III. Control of host resistance to drug-resistant L1210 sublines by H-2-linked and non-H-2-linked genes. AB - The effects of the host's genetic composition on the host's resistance to the DBA/2 L1210 lymphoma and three drug-resistant L1210 sublines (which show an increased expression of tumor-associated antigens) were investigated. Histocompatible F1 mice between DBA/2 and other mouse strains were given injections ip of graded numbers of these tumor cells, and a marked difference in the resistance of the host to tumor growth was demonstrated. The resistant hybrid mice showed markedly higher resistance to the L1210 sublines than to the L1210 parent lymphoma cells. Experiments with F1 mice between DBA/2 (H-2d) and certain congeneic intra-H-2 recombinant strains showed that the H-2b/d heterozygosity at the H-2K+l-A regions and additional non-H-2 genetic factor(s) confer the resistance to the F1 mice. The resistance was abolished by treating host animals with injections of silica, an antimacrophage agent, or rabbit anti-mouse thymocyte serum. These results suggest that the resistance is immunologic and that the genetically controlled host resistance may be directed to surface changes related to the increased expression of tumor-associated antigens. PMID- 6296516 TI - Natural killer cells in relation to Herpesvirus saimiri-induced disease course and manifestations in owl monkeys. AB - Infection of owl monkeys with Herpesvirus saimiri (HVS) results in disease courses ranging from chronic infections to fatal cancers. Malignant disease manifestations include the appearance of transformed T-cells, suppressor T-cells, and virus-carrying cells in the peripheral circulation. The reasons for the variability in disease course remain unknown. This study examined natural killer (NK) functions in relation to disease manifestations following HVS infection. The results from lymphocyte fractionation studies indicated that the owl monkey NK cell was of T-cell lineage and had receptors for the Fc fragment of IgG. Following virus infection, NK activity was enhanced in parallel with the appearance of transformed and suppressor cells in monkeys that developed malignant disease. Further studies on the relationship of these three disease characteristics indicated that they involved at least partially different T-cell subpopulations. The results further indicated that the different disease manifestations induced by HVS were polyclonal and at least partially independent events. PMID- 6296517 TI - Separation on Percoll density gradients of cells derived from malignant ascites of mice. AB - In an attempt to separate malignant from normal and reactive stromal cells, we fractionated ascites cells from BALB/c mice bearing a transplantable myeloma (MPC 11) by isopyknic centrifugation in continuous density gradients of povidone coated silica gels (Percoll). Cells from different fractions were then analyzed by morphologic and immunologic criteria. The ability of cells from the different fractions to form colonies in soft agar and to produce tumors in BALB/c mice was also examined. Although most fractions contained morphologically identifiable plasma cells, colony-forming cells (CFC), derived from multiply passaged tumors, separated in a sharp peak at 1.072 g/ml. CFC peaked at 1.078-1.082 g/ml for tumors passed less than three times and were invariably markedly depleted from the low-density portions of the gradients. Cells recovered from different fractions of the gradients were cultured in soft agar and inoculated sc into syngeneic mice. In these experiments, a highly significant correlation was observed between the ability of cells to form colonies in soft agar and to form tumors in vivo. This correlation suggests that CFC and tumorigenic cells have similar distributions. PMID- 6296518 TI - Survey of thorotrast-associated liver cancers in Japan. AB - Data on 93 autopsy cases (group A) of thorotrast-associated liver cancers were obtained from the "Annual of Pathological Autopsy Cases in Japan" from 1958 to 1979, and data on 78 autopsy cases (group B) of thorotrast-associated liver cancers were obtained from the Japanese literature from 1953 to 1980. Cholangiocarcinoma (CLC) constituted 58% of group A and 55% of group B. The curve of the cumulative numbers of patients with CLC versus year in group A was almost linear, showing an increasing risk per surviving patients with advancing time. Angiosarcoma (AGS) occurred in 25% of group A and 24% of group B. The number of patients with AGS increased significantly after 1969 in both groups (P less than 0.05). In group B, age and years after thorotrast injection of patients with AGS were statistically higher than those of patients with CLC (age: P less than 0.05; years after thorotrast injection: P less than 0.0001). Hepatocellular carcinoma (HPC) accounted for 17 and 21% of groups A and B, respectively. When yearly distribution, age, and time after thorotrast injection of patients with HPC were correlated with those of patients with other liver cancers, the only statistically significant difference between patients with HPC and patients with CLC (P less than 0.02) was in the years after thorotrast administration. Since 1977 multiple primary liver cancers including AGS developed in thorotrast administered patients in both groups. PMID- 6296519 TI - Enhancement of murine mammary tumorigenesis by feeding high levels of dietary fat: a hormonal mechanism? PMID- 6296520 TI - Naturally occurring cell-mediated cytotoxicity against simian virus 40 transformed 3T3 murine tumor cells. AB - The tumorigenicity and host protective mechanisms induced by simian virus 40 (SV40)-transformed 3T3 cells (SV403T3) were evaluated in syngeneic BALB/c mice. Tumors were regularly produced by sc inoculation of SV403T3 cells; the incidence, latent period, and survival were proportional to the size of the initial inoculum. With the use of an in vitro 18-hour 51Cr cytotoxicity assay, spleen cells from normal mice showed a dose-related killing activity against the SV403T3 cells. At an effector cell-to-target cell ratio of 200:1, the average lysis was 56 +/- 6%. This reaction appeared specific for the virally transformed targets; the mean lysis of parent 3T3 cells was 23 +/- 5%. Effectors were resistant to anti-theta serum and not removed by adherence to plastic or nylon wool. Tissue distribution studies indicated that these effectors were present in high concentrations in spleen, bone marrow, lymph nodes, and peritoneal cavity. Low levels of activity were associated with cells from the thymus. In the present studies specific T-cell cytotoxicity against the SV403T3 cells could not be demonstrated. Animals challenged with nonviable SV403T3 cells prior to tumor cell inoculation did not show increased in vivo resistance. In parallel, the in vitro cytotoxicity of animals inoculated with SV403T3 tumor cells showed no heightened cell killing compared to the cytotoxicity of normal controls. PMID- 6296521 TI - Characterization of tumor-specific transplantation antigen induced in cells infected or transformed by polyomavirus. AB - Tumor-specific transplantation antigen (TSTA) activity was investigated (with the use of inbred BALB/c and C3H mice and inbred Fisher and Wistar rats) in various early polyomavirus (Py)-coded proteins by three methodologies: 1) immunoprecipitation by anti-T-antigen sera, gel slicing, and immunization; 2) affinity chromatography with the use of anti-tumor-associated antigen serum; and 3) use of a Py mutant, NG18. The results obtained by all three techniques allowed refutation of the hypothesis that the three major species of T-antigen possess the main TSTA activity, particularly middle-sized T-antigen which is a plasma membrane protein. TSTA activity was found to be associated with proteins having a molecular weight of approximately 37,000. The exact origin of this protein is discussed. PMID- 6296522 TI - Thick ascending limb of Henle's loop. AB - Thick ascending limbs of Henle's loop have at least three major roles: (1) They reabsorb sodium chloride which dilutes the urine. (2) The reabsorption of sodium chloride also produces concentration gradients that drive the countercurrent multiplier system in the medulla and medullary rays and thus concentrates the urine. (3) They reabsorb large amounts of potassium, calcium, and magnesium in an energy-efficient manner. The mechanisms involved in these functions are reviewed in this paper, emphasizing the results of direct studies on isolated tubule segments. PMID- 6296523 TI - Acidification in the in vitro perfused tubule. PMID- 6296524 TI - Expression of ontogeny in individual nephron segments. PMID- 6296525 TI - Antiviral activity of IgG subclasses in guinea pig antiserum. PMID- 6296526 TI - [Polyglycolic acid sutures in cataract extraction]. PMID- 6296527 TI - [Connatal endocrine disorders caused by receptor defects]. AB - Since the description in 1942 of pseudohypoparathyroidism by Albright et al. hormone resistances have been documented for several peptide and steroid hormones as well as the thyroid hormones. Some of these diseases appear to be acquired while others are genetic. The hormone unresponsiveness may be complete or partial and may involve multiple hormone systems, as demonstrated in pseudohypoparathyroidism type I. Hormone resistances are characterized by hypofunction of hormone systems combined with a normal or even high secretion of the particular hormones. The connatal endocrinopathies where receptor defects have been implicated are reviewed. PMID- 6296528 TI - [Irreversible generalized pulmonary emphysema resulting from destructive bronchitis and bronchiolitis following adenovirus infection]. AB - Report of an 1 1/2 year old boy with the typical features of Williams-Campbell syndrome: chest deformity (thorax piriformis), pulmonary hyperinflation, ballooning of the segmental and subsegmental bronchi during inspiration and collapse with expiration. Atelectasis of the left upper lobe after bronchiolitis obliterans. The literature reports 23 patients with Williams-Campbell syndrome. There is every reason to believe that the extensive bronchiectasis are not due primarily to developmental deficiency of the bronchial cartilage, as the most authors assume, but result from destructive changes after inflammation. In our case very likely an adenovirus infection was the cause. The clinical features of the syndrome look like a severe therapy resistant asthma bronchiale. PMID- 6296529 TI - [Antibodies to hepatitis A and B in healthy children, mentally retarded children in institutions, and in employees of a children's hospital]. PMID- 6296531 TI - [Don't try to stop death. Care of tumor patients from the viewpoint of the clergy]. PMID- 6296530 TI - [Functional state of the pituitary-adrenal system in malignant lung neoplasms]. PMID- 6296532 TI - [The "uninteresting case" in the hospital]. PMID- 6296533 TI - [Efficient decubitus therapy by standardization using 5 principles of therapy]. PMID- 6296534 TI - [When students turn to the administrative adviser: St. Galler now also gets inconvenience allowance. Report of a political action]. PMID- 6296535 TI - [Readers' letters. Once more part-time work]. PMID- 6296536 TI - [Uppsala 1982 or research in nursing care is alive and well]. PMID- 6296537 TI - [Our experiences concerning Op-site dressings]. PMID- 6296538 TI - [Role of the public health nurse in the CLSAD (Lausanne Center of Home Care Services)]. PMID- 6296539 TI - Experimental influenza B virus toxicity in mice. A possible model for Reye's syndrome. AB - Mice intravenously injected with concentrated infectious influenza B/Lee/40 virus (LD50 = 6400 hemagglutinin units) developed lethargy, seizures, coma, and death 1 to 3 days later. The cerebrospinal fluid cell count was normal. Serum glutamic oxaloacetic transaminase levels increased 19-fold and plasma ammonia levels elevated 2.6-fold over control values. Serum bilirubin levels remained normal. Microvesicular fatty metamorphosis developed in the liver, whereas the brain showed mild cerebral edema without inflammatory changes. Viral propagation did not occur in liver or brain, but viral hemagglutinin, neuraminidase, and probably nucleoprotein antigens were produced in hepatocytes. Many of the clinical, biochemical, and pathologic features of the mouse illness are similar to those seen in Reye's syndrome. PMID- 6296540 TI - Decreased myocardial calcium uptake after isoproterenol in streptozotocin-induced diabetic rats. Studies in the in vitro perfused heart. AB - Myocardial calcium uptake in response to isoproterenol (ISO, 10(-4) M) was investigated in control and streptozotocin-induced diabetic rats using an in vitro heart perfusion model. An initial labeling with 45Ca added to the perfusate (with or without ISO) was followed by a cold calcium-free washout, thus clearing the myocardial interstitium of 45Ca. In this way the remaining 45Ca was an estimate of the myocardial calcium uptake during the labeling period. In control rats ISO induced a statistically significant increase in myocardial calcium uptake within the first 5 minutes, as well as from the 5th to the 15th minutes after exposure to this strong beta-receptor agonist. In contrast to this, diabetic hearts showed no increase in calcium uptake during any of these periods. The toxic effect of ISO was expressed by a leak of creatinine phosphokinase to the perfusate. In control rats the concentration of creatinine phosphokinase increased after ISO with a statistically significant correlation to the calcium uptake, whereas no enzymatic leak was seen after perfusion of the diabetic hearts. This abnormal response to strong beta-receptor stimulation in experimental diabetes is in accordance with our earlier in vivo finding of a myocardial protection against toxic doses of ISO. These results indicate a decreased catacholamine-induced calcium transport through the myocardial sarcolemma in streptozotocin-induced diabetic rats. This might hypothetically have relevance for diabetic heart disease as well as diabetic neuropathy. PMID- 6296541 TI - Experimental studies on malignant fibrous histiocytomas. II. Ultrastructure of malignant fibrous histiocytomas induced by 4-(hydroxyamino)-quinoline 1-oxide in rats. AB - The ultrastructures of six subcutaneous and six bone malignant fibrous histiocytomas (MFH) induced in rats by local application of the carcinogen, 4 (hydroxyamino)-quinoline 1-oxide (4-HAQO) were studied. The MFHs could be classified histologically into three subtypes: of the six subcutaneous MFHs, four were fibrous, one was giant cell, and one was myxoid; of the osseous MFHs, three were fibrous, one was giant cell, and two were myxoid. Five different types of cells were found in the MFHs: fibroblast-like cells, histiocyte-like cells, undifferentiated cells, xanthomatous cells, and multinucleated giant cells; the xanthomatous cells and multinucleated giant cells, however, were probably derived from histiocyte-like cells. Fibroblast-like cells predominated in storiform areas of the fibrous subtype; histiocyte-like cells and undifferentiated cells predominated in the giant cell subtype; intermediate cells predominated in the myxoid subtype. Acid phosphatase activity was found in lysosomes and myelin figures of the histiocyte-like cells in fibrous type MFH. The giant cell subtype of bone MFH has been transplanted serially into syngeneic rats and is now at the 17th generation. Transplantability exceeded 80%; doubling time was 3.8 to 6.1 days. Until the 3rd generation, the histology of the original tumor was retained; from the 4th generation, however, giant cells and xanthoma cells were no longer observed, and the tumor was composed mainly of undifferentiated cells. These results indicate that (a) MFH induced in the rat by 4-HAQO have an ultrastructure similar to human MFH and (b) the giant cell subtype transplanted serially is gradually transformed with a probable selection of stem cells and undifferentiated cells. PMID- 6296542 TI - Hepatomas in young women on oral contraceptives: report of two cases and review of the literature. AB - Hepatocellular carcinoma (HCC) developed in two women who had ingested oral contraceptives for prolonged duration. One of the patients had hepatoma restricted to the liver, and it was successfully resected. The other patient presented with metastatic HCC to the sacrum and lung, and she died after an initial response to chemotherapy in 12 months. Review of literature showed 23 previously reported cases of HCC associated with "pill" use since 1973. Although the reported instances of HCC in pill users are few in number, the majority of affected females were in their 20s and early 30s, without any preexisting liver disease. Even though the risk of developing HCC on oral contraceptives is low, this association adds to the list of complications induced by oral contraceptives in premenopausal women. Two cases are reported and literature reviewed. PMID- 6296543 TI - Extrarenal Wilms tumor. PMID- 6296544 TI - Synchronous bilateral primary germ cell tumors of the testis: a case report and review of the literature. AB - Synchronous bilateral primary germ cell tumors of the testis are exceedingly rare. The most common synchronous testicular tumors are seminomas, followed by embryonal carcinomas, teratocarcinomas, and choriocarcinomas. In a series of 385 patients we found nine with bilateral primary germ cell tumors of the testis (2.3%), including one with synchronous involvement of both testes. The treatment of synchronous bilateral primary germ cell tumors of the testis is in principle the same as that of solitary testicular primary germ cell tumors, and is based on tumor histology and tumor metastasis. PMID- 6296545 TI - Primary adenocarcinoma of the appendix with bilateral Krukenberg ovarian tumors. AB - An unusual case of a 22-year-old white female with known chronic ulcerative colitis presented with a several-day history of lower abdominal pain and a pelvic mass. Laparotomy revealed a primary carcinoma of the appendix with Krukenberg metastasis to both ovaries. Pathologically this tumor appeared to arise from an appendix which showed no evidence of chronic ulcerative colitis and therefore could not be associated with the above-mentioned entity. PMID- 6296546 TI - Adriamycin, CCNU, and 5-fluorouracil in patients with advanced gastrointestinal cancer. AB - Forty-one patients with measurable advanced gastrointestinal malignancy were treated with a combination of Adriamycin (doxorubicin), 5-FU (5-fluorouracil), and CCNU (1-(2-chloroethyl)-3-cyclohexyl-1-nitrosourea). The response to therapy was evaluated every eight weeks. In addition to standard clinical examinations and laboratory tests to evaluate toxicity, 31 patients were asked to rank both the expected and experienced negative effects of the treatment. Although both patients with hepatocellular cancer and the patient with a soft tissue sarcoma responded to the regimen, only 1 of 38 patients with adenocarcinoma had a favourable response. The clinically measurable toxicity, principally to the gastrointestinal tract and bone marrow, was relatively mild. However, the patients' subjective evaluation of their treatment made the side effects appear more significant. The results suggest that this regimen deserves further evaluation in patients with hepatocellular cancer and soft tissue sarcomas. PMID- 6296547 TI - Alternating multiagent chemotherapy for advanced ovarian cancer. AB - A chemotherapy regimen consisting of hexamethylmelamine (H) 150 mg/m2 orally days 1-14, cyclophosphamide (C) 500 mg/m2 IV day 1 of a 28-day cycle with Adriamycin (A) 40 mg/m2 IV day 1 alternating with cis-diamminechloroplatinum (C-P) 50 mg/m2 IV day 1 every other cycle was administered to 29 patients with advanced epithelial ovarian cancer. Toxicity to this regimen included alopecia, nausea, and vomiting in all patients. Mild paresthesias occurred in four patients. Hematologic toxicity required only minimal dose modification. There was no cardiac, renal, or auditory toxicity. The clinical response rate of 55% and median survival of 14 months compare favorably with that of other reported series. This chemotherapy regimen seems to be well tolerated without jeopardizing the patients' response. PMID- 6296548 TI - Development and use of a nonrestraining waveguide chamber for rapid microwave radiation killing of the mouse and neonate rat. AB - The use of microwave energy for rapid killing of small rodents such as the mouse or rat has become a standard pharmacologic technique since approximately 1975. This method allows investigation of rapidly modulated neurochemical indices, neuromodulatory substances, and some neurotransmitters to be determined at basal concentrations in brain regions and microregions. Previously described devices for use with microwave generators have relied on total body restraining holders in order to properly position rodents and neonates within a closed waveguide during microwave energy exposures. The present information describes two alternate chamber designs which do not require restraint of the rodent. A positioning device is described which must be used with the waveguide chambers. The animal chambers are designed to be used with 2450 MHz energy. PMID- 6296549 TI - An electrostatic model for mechano-electrical transduction, especially in the ear. PMID- 6296550 TI - Establishment and characterization of a human EBV-negative B cell line (MN 60). AB - A permanent cell line, MN 60, was established from the peripheral blood of a patient with an acute lymphoblastic leukemia (ALL) classified morphologically as being of the L3 type. Cell growth started rapidly in vitro and no feeder cells were needed. Cells of the MN-60 line were identical to the original leukemic cells with respect to surface immunoglobulin (Ig) expression and karyotype, including the presence of four marker chromosomes [1q+, 6q-, t(8;14)]. Continuous proliferation was maintained in stationary suspension culture with a doubling time of 25 h. The cells were tumorigenic in athymic nude mice and had the capacity to form colonies in semi-solid medium in vitro. Monoclonal surface Ig (mu lambda) was demonstrated whereas no cytoplasmic immunoglobulin could be demonstrated. The MN-60 cells were Epstein-Barr virus (EBV) negative as evidenced by EBNA tests and by nucleic acid hybridization studies. The cells expressed HLA A-C, HLA-DR. beta 2-Microglobulin and cALL, but not Fc gamma. C3, sheep and mouse red blood cell receptors. No reactivity was found with anti-glycophorin A or the anti-BL 38.13 monoclonal antibody. Cell growth was retarded in the G0/G1 phase of the cell cycle after incubation with leukocyte interferon, hydrocortisone, phorbol myristate acetate and dimethyl sulphoxide. PMID- 6296551 TI - Immunodeficiency and reticulum cell sarcoma in mice segregating for HRS/J and SJL/J genes. AB - Hairless (hr/hr) mice segregating for SJL/J and HRS/J genes (SJL-HRS) were compared to their haired counterparts with respect to immune responsiveness, tumour development and ecotropic murine leukemia virus (MuLV) expression. Homozygosity at the hairless locus did not affect expression of MuLV. There was however, a significant depression of the cellular immune response of these mice as characterized by depressed reactions in phytohemagglutinin, concanavalin A and mixed leukocyte assays. Haired and hairless mice did not differ significantly in response to B-cell mitogens or in production of cytotoxic antibody. The depressed cellular immune response in hr/hr mice is associated with a distinctive histologic type of spontaneous reticulum cell sarcomas. The importance of these results in relation to previous studies of HRS/J hairless mice is discussed. PMID- 6296552 TI - Establishment and characterization of a new permanent cell line (GDM-1) from a patient with myelomonoblastic leukemia. AB - The GDM-1 permanent cell line was established from the peripheral blood of a patient with a Philadelphia chromosome negative myeloproliferative disorder, after transformation to acute myelomonoblastic leukemia. The GDM-1 cells exhibited the same characteristics as those isolated from the peripheral blood of the patient prior to death: cells contained non-specific esterase sensitive to fluoride, myeloperoxidase, lysozyme (muramidase), and exhibited both Fc and complement (C3) receptors but lacked B- and T-cell surface markers including T associated antigens. E-rosetting capacity, surface and intracytoplasmic immunoglobulins and EBV determined nuclear antigen (EBNA). The GDM-1 cells bore the 1a receptor and the myeloid leukemia antigen (M-1). The karyotype of the cultured leukemic cells showed the same specific chromosomal abnormalities present in the monoblasts obtained from the peripheral blood prior to death, indicating that the cell line was derived from the original leukemic cells. PMID- 6296553 TI - Terminal deoxynucleotidyl transferase (TdT) in RadLV-induced C57BL thymomas. AB - Newborn C57BL mice exposed to RadLV at birth were followed and periodically sampled for thymus and bone marrow TdT activity. Tumors detected at or before 100 days had mostly low TdT levels. Large thymic lymphocytic tumors detected after 100 days had elevated total TdT content per gland. Serial samples showed a changing TdT pattern with time. The mid period shifted from low levels to normal to-high levels for both TdT content and total TdT per gland. No unusual bone marrow TdT activity change preceded the appearance of thymomas. TdT marks a unique precursor cell highly sensitive to RadLV associated with lymphomagenesis. RadLV induced both TdT+ and TdT- thymic lymphomas and both types were found to occur after lymphoma initiation by injection of RadLV into newborns. PMID- 6296554 TI - Neodymium-YAG laser therapy of malignant airway obstruction. A preliminary report. AB - Neodymium-YAG laser therapy of malignant obstructing airway lesions via a flexible endoscope offers a promising new means of treatment. Twenty-two patients with obstructing airway lesions were treated. Two fatal complications occurred. The patients experienced increased comfort in breathing, increased exercise tolerance, and more effective cough with clearance of secretions. Treatment of larger numbers of patients with careful monitoring of the results should be performed to define further the usefulness, complications, and limitations of this new therapeutic tool. PMID- 6296555 TI - Angiotensin-converting enzyme in sarcoidosis. PMID- 6296556 TI - Specificity of monoclonal antibodies to an EBV transformed B-cell line. AB - Monoclonal antibodies were produced against an Epstein Barr virus (EBV) transformed human B-cell line with the following HLA specificities: HLA A2, B27, Cw2, Dr3,2. Antibodies from three clones, Mab B1, Mab B2 and B3 reacted with human Ia-like molecules on peripheral blood B cells, some monocytes, CLL cells, lymphoblastoid B-cell lines and some mixed leukocyte culture (MLC)-activated T cells, but were unreactive with leukoagglutinin (LA) and Concanavalin A (Con A) activated T blasts and T-cell lines. Antibodies obtained from three other clones (Mab 4, 5 and 6) reacted with a mw 80,000 protein present on peripheral lymphocytes and on most of the lymphoblastoid T-and B-cell lines tested. PMID- 6296557 TI - Calcium-dependent increase in the potassium permeability of human red blood cells by pentachlorophenol and 2,4,6-trinitrophenol. AB - The effects of four phenol derivatives on the Ca2+ and K+ permeability of human red cells were studied at 0 degree C using 45Ca2+ and 86Rb+. Pentachlorophenol (PCP) and 2,4,6-trinitrophenol (TNP), which are mainly ionized at physiological pH, produced a marked influx of Ca2+ that increased with increasing pH. The elevated intracellular Ca2+ concentration was completely (TNP 5-10 mM, PCP 0.5 mM) or partly (PCP 1.0 mM) responsible for a concomitant increase in potassium permeability. No effects were found in connection with the weakly ionized 4-CI and 4-CI-3,5-di-CH3 derivatives at 1.0-5.0 mM and 0.5-2.0 mM, respectively. The fact that ionized phenols are able to transfer Ca2+ from water to a hydrophobic phase suggests that they may act as weak Ca2+ ionophores in the red cell plasma membrane. PMID- 6296558 TI - [Dehydroepiandrosterone sulphate (DHAS) levels during childhood and adolescence]. PMID- 6296559 TI - [An evaluation of sputum cytology in the diagnosis of bronchogenic carcinoma]. PMID- 6296561 TI - [Anaplastic small cell carcinoma of the lung as a cause of acquired stenosis of the pulmonary artery. Pulmonary emboli as initial manifestations]. PMID- 6296562 TI - [Focal dermal hypoplasia. Goltz syndrome]. PMID- 6296560 TI - [Cis-dichlorodiammineplatinum, adriamycin and cyclophosphamide versus cis dichlorodiammineplatinum, adriamycin, cyclophosphamide and CCNU in non-small cell carcinoma of the lung]. PMID- 6296563 TI - [Glomoblastoma. Bibliographic review and presentation of a clinical case]. PMID- 6296564 TI - Tracheal involvement in laryngeal papillomatosis. AB - A review of 39 laryngeal papilloma patients at The Johns Hopkins Hospital revealed an incidence of subglottic and tracheal extension much higher than previously reported. Subglottic disease was present in 69% of patients and tracheal involvement in 26%. Factors associated with distal spread were the presence of subglottic papillomas, history of tracheostomy, total number of endoscopic resections, and the duration of disease. The relationship to the presence of viral antigen was cited. Detection and elimination of all papillomas is felt to be the key for the prevention of distal spread. The strategy of management is described. PMID- 6296565 TI - Malignant fibrous histiocytoma in the maxillary sinus. Xanthoma-like change of the tumor after radiotherapy. AB - A case of malignant fibrous histiocytoma (MFH) in the right maxillary sinus is reported. Patient was a 30-year-old male, and was diagnosed as pleomorphic type of MFH by biopsy before therapy. The tumor shrunk and changed into xanthoma-like tumor after radiotherapy. A case like the present case has not been reported previously in the literature. PMID- 6296566 TI - Proceedings of the 1982 International Narcotic Research Conference. Part II. PMID- 6296567 TI - Receptor-related interactions of opiates with PGE-induced adenylate cyclase in brain. AB - The inhibition by opiates of the PGE2-induced formation of cAMP in slices from rat brain striatum was investigated. A maximal, 3.5-fold increase over the basal level of cAMP was obtained with an EC50 for PGE2 of 3 microM. Opiate agonists of both mu and kappa type were inhibitory. The IC50 values for morphine, levorphanol and ethylketocyclazocine (EKC) were 110 nM, 80 nM and 25 nM, respectively. These values were similar to the potencies of the compounds in displacing stereospecifically bound 3H-etorphine in rat brain membranes. As evidenced by the inactivity of dextrorphan, the inhibition of PGE2-dependent cAMP formation was stereospecific. Also ineffective were the opiate antagonists naloxone, naltrexone and MR 2266. These compounds did, however, reverse the inhibition by agonists, displaying thereby selectivity toward the putative mu and kappa opiates. Thus, the inhibition by morphine was antagonized to a greater degree by naloxone than by MR 2266, and the action of EKC was blocked more effectively by MR 2266 relative to naloxone. PMID- 6296568 TI - Partial loss of opiate receptor regulation by GPP(NH)P evident in vitro. PMID- 6296569 TI - Photolabile ligands for opiate receptors. AB - The 2-nitro-4-azidophenyl(NAP)-D-Ala2-Leu5-Enkephalin derivatives: Try-D-Ala-Gly Phe-Leu CONCH2CH2NH-NAP (E-NAP-EDA) and Try-D-Ala-Gly-Phe-Leu CONCH2CH2NH COCH2CH2NHNAP(E-NAP- -Ala-EDA) were synthesized by conventional peptide methods. Their structure was determined by amino acid analysis, ultra violet, visible and infra red spectroscopy. Both peptides were shown a) to bind with high affinity to the opiate receptors of rat brain membranes and b) to inhibit strongly the contractions of electrically stimulated vas deferens and the adenyl cyclase of the NG 108-15 cell membranes. These effects were reversed by the antagonist naloxone. Photoloysis of the rat brain membranes-(E-NAP- -Ala-EDA) complex caused a 20-30% inactivation of the opiate receptors. Inactivation was prevented when the complex was irradiated in the presence of naloxone. The radio-labeled derivatives of these enkephalin analogs may prove useful photochemical labels of the opiate receptor. PMID- 6296570 TI - Characterization of type 2 opiate receptors. AB - Recent evidence suggests that the Type 1 opiate receptor (in rat striatal patches) is a mobile receptor which is able to adopt a mu, delta, or kappa opiate receptor ligand selectivity pattern under appropriate conditions. In this paper, we have investigated such a possibility for Type 2 opiate receptors which are visualized diffusely over rat striatum. Ligand selectivity analysis suggested that the Type 2 opiate binding site is equivalent to a delta opiate receptor. The auto-radiographic distribution of Type 2 opiate binding sites is diffuse over most areas of rat brain. Thus, Type 2 opiate binding sites are different from Type 1 opiate receptors which are very discretely distributed in rat brain. Our results suggest that Type 2 opiate receptors, unlike Type 1 opiate receptors, are receptors locked in a delta-like ligand selectivity conformation. PMID- 6296571 TI - Nigral 6-hydroxydopamine lesions equally decrease mu and delta opiate binding to striatal patches: further evidence for a conformationally malleable type 1 opiate receptor. AB - We have investigated the effect of nigral 6-hydroxydopamine (6-OHDA) lesions on binding of the mu receptor ligand dihydromorphine (DHM) and the delta receptor ligand [D-Ala2, D-Leu5]-enkephalin (DADLE) to sections of rat striatum under conditions which yield mu-like and delta-like ligand selectivities at discrete receptor patches (Type 1 receptor). 3H-DHM binding was decreased 43% while 3H DADLE was decreased 22%. However, when the contribution of diffuse binding (Type 2) which is not affected by 6-OHDA is subtracted from the patch, the decrease is approximately 49% for both ligands. These data support the hypothesis that the Type 1 receptor of striatal patches is a conformationally malleable receptor entity which can exist in states having high affinities for various classes of opiate ligands. PMID- 6296572 TI - Reduction in opiate receptor reserve in morphine tolerant guinea pig ilea. AB - Spare opiate receptors in the guinea pig ileum have been detected by the use of the opiate receptor alkylating agent beta-chlornaltrexamine (CNA). Treatment of the guinea pig ileum longitudinal muscle in vitro with low concentrations (less than 10nM) of CNA resulted in an irreversible parallel shift to the right of the normorphine log concentration response curve. With increasing concentration of the reagent, the agonist EC50 becomes progressively greater. Finally a point is reached at which the maximal agonist effect decreases, so that parallelism is no longer seen. The maximal parallel shift provides a measure from which one can estimate the spare receptor fraction that is present in untreated tissue. In ilea from normal guinea pigs, roughly 80-90% of the opiate receptors for normorphine were found to be spare. Even after the largest parallel shifts that could be achieved, the naloxone Ke value for antagonism was unchanged, indicating that normorphine acts through spare mu receptors. Ilea from guinea pigs made tolerant by chronic morphine pellet implantation were found to be more sensitive to the effects of CNA treatment; there was a reduction in the number of spare receptors for normorphine. It is suggested that the opiate spare receptor fraction is physiologically modulated to control neuronal sensitivity to opioid effect. PMID- 6296573 TI - Prototype cannabinoid analgetics, prostaglandins and opiates--a search for points of mechanistic interaction. PMID- 6296574 TI - Characteristics of a monoclonal beta-endorphin antibody recognizing the N terminus of opioid peptides. AB - The properties of a mouse monoclonal antibody to beta-endorphin secreted by a clone of hybrid myelomas (3-E7) are described. The antibody displays virtually complete cross-reactivity to met-enkephalin and leu-enkephalin, but no cross reactivity to beta-lipotropin, alpha-N-acetyl-beta-endorphin and des-Tyr1-beta endorphin. Substantial cross-reactivity is seen with some other naturally occurring opioid peptides bearing the enkephalin sequence, such as dynorphin, alpha-neo-endorphin and BAM 22, but cross-reactivity is lacking in the case of certain synthetic enkephalin derivatives possessing a D-amino acid in position 2. The data indicate that for the binding of an antigen to the antibody the N terminal tyrosine moiety is essential. The antibody recognizes, thus, a site which is of functional significance for the interaction of many naturally occurring opioid peptides with the opiate receptor. PMID- 6296575 TI - New enkephalinase inhibitors as probes to differentiate "enkephalinase" and angiotensin-converting-enzyme active sites. PMID- 6296576 TI - Enkephalin-hydrolyzing peptidases of rat brain membranes: are they topographically/functionally coupled to opiate receptors? PMID- 6296577 TI - Plasma measures of B-endorphin-like immunoreactivity in depressives and other psychiatric subjects. PMID- 6296578 TI - Plasma beta-endorphin-like immunoreactivity, self reported pain perception and anxiety levels in women during pregnancy and labor. AB - Plasma Beta-endorphin (B-END) immunoreactivity was measured in 19 men, 16 women at midcycle, and ten pregnant women at various points in their pregnancies. There is a significant difference between the levels measured in males and females (t = 3.74, df = 31, p = .0007). Pregnant women demonstrated a steady increase in plasma Beta-End-like immunoreactivity between second and third trimester and through labor. The levels dropped rapidly postpartum. The material being measured is predominantly B-end-sized. Psychological studies indicate that these changes are not strongly correlated with pain perception or self-reported anxiety levels. PMID- 6296579 TI - alpha, N-acetyl derivatives of beta-endorphin in rat pituitary: chromatographic evidence for processed forms of beta-endorphin in pancreas and brain. PMID- 6296580 TI - Changes in high affinity sodium independent gamma-aminobutyric acid binding in cerebral cortex and hippocampus of the rat following electroshock. AB - Electroshock induced seizures in the rat enhanced high affinity specific Na+ independent binding of 3H-gamma aminobutyric acid (GABA) to frozen, Triton X-100 treated cerebral synaptic membranes 30 minutes after exposures to electroshock, although no change in 3H GABA binding was observed in similarly treated preparations from hippocampus. Scatchard analysis of the binding isotherms from cortical membranes indicated that the increase in 3H GABA binding at 30 minutes was due to a rapid increase (39%) in the number of available GABA receptor binding sites (Bmax) rather than an alteration in receptor affinity (KD). The number of binding sites returned to control values within 1 hour and remained so throughout the duration of this study. PMID- 6296581 TI - Loss of rat ventricular alpha 1-adrenergic receptors during aging. AB - Ventricular alpha 1-adrenergic receptor concentration, measured by specific binding of [3H]-prazosin, decreases by 33% as rats age from 3 to 24 months. No age changes occur in binding affinity for [3H]-prazosin or potency of various alpha-adrenergic agonists and antagonists to displace [3H]-prazosin. The ratio of membrane protein to ventricular wet weight also does not change significantly with age. These results suggest a possible mechanism for loss of cardiovascular alpha-adrenergic responsiveness during aging. PMID- 6296582 TI - Effects of insulin-glucagon interactions on glycogenolysis and protein kinase activity in rat hepatocytes. AB - The effects of insulin and glucagon on cAMP accumulation, protein kinase activation, and glycogenolysis were investigated in isolated rat hepatocytes. Glucagon (0.01 nM to 10 micro M) increased the activation state of protein kinase and the rate of glucose accumulation. Addition of 1.0 nM insulin to cells preincubated with 0.1 nM glucagon attenuated the rate of glucose accumulation, but did not alter the protein kinase activity ratio. Addition of 0.1 nM glucagon to cells preincubated with 1.0 nM insulin caused a rapid activation of protein kinase; however, glycogenolysis was not immediately affected. These effects were enhanced with pharmacological concentrations of glucagon and insulin. These data indicate that the degree of protein kinase activation does not always correlate temporally or quantitatively with rates of glycogenolysis in liver cells exposed to insulin and glucagon. PMID- 6296583 TI - The roles of opioid peptides in controlling thyroid stimulating hormone release. AB - We have studied the role of the opioid peptides in controlling TSH secretion. Morphine sulfate significantly decreased, while naloxone had no effect on, basal plasma TSH levels of female rats. In contrast, naloxone blocked the stress induced fall in plasma TSH. Microinjection of beta-endorphin into the third ventricle resulted in a fall in TSH while such injection of naloxone into the posterior hypothalamus increased TSH. Microinjection of beta-endorphin directly into the pituitary caused a rise in plasma TSH. It is concluded that opioid peptides probably play no role in basal TSH secretion, but are involved in the stress-induced fall in TSH. Furthermore, it appears that opioid peptides have a site of action in the hypothalamus to decrease TSH and a direct pituitary action to increase TSH. PMID- 6296584 TI - Tricyclic antidepressant desipramine induces stereospecific opiate binding and lipid modifications in rat glioma C6 cells. AB - Incubation for 48 hours of C6 glioma cell cultures with 10(-4)M tricyclic antidepressant desipramine gave rise to a quantitative increase of total lipids and to qualitative modifications of glycosphinegolipids involving detection by thin-layer chromatography of spots migrating according to cerebroside and sulfatide and presence of an abnormal ganglioside pattern. These lipid modifications were associated with the appearance of stereospecific binding of opiates (dihydromorphine) with a dissociation constant of 30-60 nM. These results favor an important role of lipids in opioid receptor function. PMID- 6296585 TI - Effects of thyrotrophin stimulation for two hours on mouse thyroid cyclic AMP levels in vivo and in vitro. AB - A thyrotrophin (TSH) stimulation in vivo in mice for 2 h, reflected by continuously increasing plasma triiodothyronine (T3) levels, was associated with an increase in the thyroid content of cyclic AMP (cAMP) during the first 25 min of stimulation; thereafter the level rapidly declined. A similar pattern of the cAMP response was found when mouse thyroid tissue was stimulated by TSH in vitro for 2 h. This is an in vivo demonstration of a type of cAMP response to prolonged hormonal stimulation that has been observed in several in vitro systems including thyroid tissue, generally referred to as hormone induced desensitization of adenyl cyclase. The present results indicate that the phenomenon is not confined to in vitro conditions but can be demonstrated also in vivo, and support the representativeness of in vitro experiments in this respect. PMID- 6296586 TI - An investigation of the role of kappa opiate receptor agonists in the initiation of feeding. PMID- 6296587 TI - Localization of human placental opiate binding sites on the syncytial brush border membrane. AB - Opiate binding sites were measured in different placental membrane fractions which were characterized by marker enzyme analysis and electron microscopic examination. The distribution pattern of opiate binding sites in the different fractions closely parallels that of placental alkaline phosphatase. These results clearly show that opiate binding sites are mainly located on the syncytial brush border membrane. The opiate binding sites found on microvillus membrane fraction have the same pharmacological characteristics as the Kappa opiate binding site previously characterized on placental crude membrane fraction. PMID- 6296588 TI - Alcohol interactions with brain opiate receptors. AB - Ethanol, added in vitro to mouse caudate membranes, inhibited high-affinity binding of 0.2 nM 3H-dihydromorphine (3H-DHM) over an ethanol concentration range of 250-1,000 mM. At lower, physiologically-attainable ethanol concentrations (e.g.: 50 mM), 3H-DHM binding was significantly increased. Over the concentration range of 50-1,000 mM, ethanol inhibited 0.5 nM 3H-[D-Ala2,D-Leu5]enkephalin (3H ENK) binding to mouse caudate tissue, and no stimulation of 3H-ENK binding was noted at any of these concentrations of ethanol. Ethanol inhibits opiate binding in a pseudo-competitive manner and, therefore, the concentration of ligand used to assess the effects of ethanol is of major importance. Results obtained with other alcohols which differ in their membrane:water partition coefficients suggest that alcohol effects on opiate binding are not solely dependent on the membrane-disordering properties of the alcohols. PMID- 6296589 TI - Benzodiazepine--specific and nonspecific tolerance following chronic flurazepam treatment. AB - Rats were given a flurazepam solution as their only water source for 4 weeks. The drug concentration was adjusted so the rats would consume 100-150 mg/kg daily. This treatment is known to cause a reduction in the number of specific benzodiazepine binding sites (receptor down-regulation) and tolerance to the locomotor impairment caused by the injection of a large test dose of flurazepam. Both the tolerance and the receptor down-regulation disappear within 24 hours after the end of chronic treatment. After 4 weeks of flurazepam treatment, rats were tested for locomotor impairment and loss of the righting response caused by pentobarbital, ethanol or diazepam. There was a small tolerance to pentobarbital. This lasted at least 4 days, but had disappeared by 7 days. Rats also had a small tolerance to ethanol, which disappeared between 24 and 48 hours after the end of chronic flurazepam treatment. In contrast, there was a large tolerance to diazepam, but this was gone by 24 hours after the end of chronic treatment. It appears that two types of tolerance develop during benzodiazepine treatment: (1) tolerance specific for benzodiazepines possibly mediated by receptor down regulation, and (2) nonspecific tolerance, possibly analogous to that which develops during chronic barbiturate treatment. PMID- 6296590 TI - Cross tolerance between morphine and beta-endorphin in the in vivo vasodilating effect in the hamster cheek pouch. AB - The relative potencies of the in vivo vasodilating effect on the arterioles of the hamster cheek pouch of morphine and beta-endorphin decreased significantly after chronic morphine injection, indicating the existence of tolerance to morphine and cross tolerance between morphine and beta-endorphin. The apparent pA2 values of both morphine-naloxone and beta-endorphin-naloxone in morphine treated hamsters also decreased significantly. The results indicate that in this preparation there is a simultaneous reduction in the affinity of the opiate receptors to both agonists as well as antagonist associated with the development of tolerance or cross tolerance. PMID- 6296591 TI - Bombesin-like peptides in small cell lung cancer: biochemical characterization and secretion from a cell line. AB - High intracellular levels of BN-like peptides are present in tumors and cell lines of small cell carcinoma of the lung (SCCL) as well as the putative precursor cells of this tumor, the pulmonary endocrine cell. In cell line NCI H209 the density of bombesin-like peptides was 8.9 +/- 1.1 pmol/mg total protein. Gel filtration chromatography of an extract of these cells revealed one major peak of immunoreactivity which coeluted with synthetic bombesin (1620 daltons). Also, high pressure liquid chromatography revealed one major peak of immunoreactivity was present which eluted before synthetic peptide. Therefore, SCCL bombesin-like peptides may be of similar size but are more hydrophilic than synthetic peptide. Cells maintained in culture continuously release bombesin-like peptides into the growth medium. Also, high concentrations of K+ stimulated the secretion of immunoreactive bombesin from cell lines in a Ca++-dependent manner. These SCCL bombesin-like peptides may function as important regulatory agents in the malignant lung. PMID- 6296592 TI - Enkephalin dimers: regulation of cyclic AMP levels in NG108-15 cells. PMID- 6296593 TI - Reversible decrease in dopaminergic 3H-agonist binding after 6-hydroxydopamine and irreversible decrease after kainic acid. AB - Six days after the unilateral intrastriatal injection of 30 ug 6-hydroxydopamine (6-OHDA) the number of stereospecific 3H-dopamine and 3H-apomorphine binding sites (Bmax) was reduced by 50-60% in the caudate nucleus ipsilateral to the lesion. The dopamine content of the lesioned caudate nucleus was also reduced to 2% of the contralateral side or of sham-operated controls. The preincubation of depleted homogenates with added dopamine reversed the effects of 6-OHDA on the Bmax of 3H-agonists. A similar pattern of depletion, decrease in binding and in vitro reversal by dopamine was observed after a single injection of reserpine (4.0 mg/kg, im.). The intrastriatal injection of kainic acid also lowered the Bmax of 3H-agonists by 65% without altering dopamine content. Preincubation of homogenates of kainic acid-lesioned caudate nuclei with 355 nM (endogenous) dopamine did not reverse the decrease in binding. We conclude that treatments which deplete endogenous dopamine, including the lesion of nigrostriatal terminals, induce a reversible change in the parameters of 3H-agonist binding whereas the destruction of intrinsic caudate neurons with kainic acid results in an irreversible loss of receptors. PMID- 6296594 TI - Synthetic corticotropin-releasing factor stimulates secretion of immunoreactive beta-endorphin/beta-lipotropin and ACTH by human fetal pituitaries in vitro. AB - Synthetic ovine CRF, in an amount approximating that found in pituitary portal plasma of the rat, induced a significant increase in the secretion of both ACTH and immunoreactive beta-endorphin/beta-LPH (i beta-END/LPH) by human fetal hemipituitaries in an in vitro superfusion system. This finding suggests that a molecule similar to synthetic ovine CRF may be a physiologic hypothalamic releasing factor in man. PMID- 6296595 TI - [Pentastomosis. A serological study of 193 Ivorians]. AB - Pentastomosis is the infection caused by the larvae of a worm resorting to Armillifer genus, the adults living in the lungs of big african snakes. Eggs are released with snakes slime and dejecta and ingested by man and various animals. Larvae are disseminated through lymphatic vessels in peritoneum, liver, spleen, lungs, pleurae, and rarely in kidneys, brain or even eye. Most often the infection is fortuitously detected during a radiologic control or a surgical inspection. The serological prevalence was of 4.2 p. 100 as a whole, somewhat lower than that given by post mortem examinations but higher than the prevalence resulting of radiologic controls. Children under 5 are concerned with a prevalence of 7 p. 100. PMID- 6296596 TI - [Clinical and epidemiologic aspects of cryptococcosis in Kinshasa. Apropos of 15 personal cases]. AB - The authors have recorded 15 cases in their eighteen months of hospital practice in Kinshasa. Fever and meningo-encephalitis are not specific symptoms and the detection of yeasts in the spinal fluid or sometimes in a lymph node is necessary to support the diagnosis. Direct examinations of the cerebro-spinal fluid is most often sufficient and easy for patients who have already had, in most cases, a long evolution giving way to an important proliferation of yeasts. Only one patient recovered by a treatment associating amphotericin B and ketoconazole. Another one, after an apparent recovery with the same treatment, died rapidly of acute tuberculosis. PMID- 6296597 TI - [Disseminated form of histoplasmosis caused by Histoplasma capsulatum in a Zairian child]. AB - American histoplasmosis, characterized by its small parasitological forms, is a visceral mycosis which though rare, has already been reported in Zaire. A new case, in a girl of 4 years is reported with bad general condition, fever and general reaction of the reticulo-endothelial system. The microscopic aspects of the yeast are given. A treatment with ketoconazole proved rapidly effective and the patient had apparently recovered when she died of measles which have in Africa and specially in fragile patients a particular severity. PMID- 6296598 TI - [Various biological parameters in sub-clinical protein-calorie malnutrition in Ivory Coast children]. AB - Protein malnutrition is the main aspect of infant pathology in the Third World. This review has been undertaken to find out some early biological indicators of nutritional deficiencies. The review, based on a sample of 1 317 children between 1 and 3 years old, has shown: --in asymptomatic protein malnutrition, hemoglobin, mean corpuscular hemoglobin, prealbumin and albumin all significantly decreased. They ease an earlier appreciation than body weight. In protein-calorie malnutrition, body weight remains the major indicator; --children weighing, associated with these parameters determinations, might improve the diagnosis of asymptomatic malnutrition. Indeed, nutrition education of mothers could be envisaged at an early stage. But quick method of determination of these parameters has still to be made practicable. PMID- 6296599 TI - [Unusual forms of X-ray pneumonitis resulting from treatment of patients with small cell bronchial carcinomas. Radiologic appearance and significance of cortisone withdrawal]. PMID- 6296600 TI - [Cefotiam in infections of granulocytopenic tumor patients]. PMID- 6296601 TI - Carbohydrate metabolism. Part E. PMID- 6296602 TI - Acetate kinase from Bacillus stearothermophilus. PMID- 6296603 TI - cAMP-dependent protein kinase, soluble and particulate, from swine kidney. PMID- 6296604 TI - Fructose-bisphosphatase, zinc-free, from rabbit liver. PMID- 6296605 TI - Fructose-bisphosphatase from ox liver. PMID- 6296606 TI - Fructose-1,6-bisphosphatase from turkey liver. PMID- 6296607 TI - Fructose-1,6-bisphosphatase from chicken and rabbit muscle. PMID- 6296608 TI - Fructose-1,6-bisphosphatase from rabbit liver (neutral form). PMID- 6296609 TI - Fructose-1,6-bisphosphatase from snake muscle (Zaocys dhumnades). PMID- 6296610 TI - Fructose-1,6-bisphosphatase from rat liver. PMID- 6296612 TI - Fructose-1,6-bisphosphatase from bumblebee flight muscle. PMID- 6296611 TI - Fructose-1,6-bisphosphatase from mouse and rabbit intestinal mucosa. PMID- 6296613 TI - Fructose-bisphosphatase from spinach leaf chloroplast and cytoplasm. PMID- 6296614 TI - Fructose-bisphosphatase from Rhodopseudomonas palustris. PMID- 6296615 TI - Fructose-1,6-bisphosphatase from Bacillus licheniformis. PMID- 6296616 TI - Sedoheptulose-1,7-bisphosphatase from wheat chloroplasts. PMID- 6296617 TI - Glucose-6-phosphatase from cerebrum. PMID- 6296618 TI - Pyruvate dehydrogenase phosphatase from bovine heart. PMID- 6296619 TI - Phosphoprotein phosphatase from swine kidney. PMID- 6296620 TI - Isolation of several abundant muscle enzymes. PMID- 6296621 TI - Simultaneous purification of glyceraldehyde-3-phosphate dehydrogenase, 3 phosphoglycerate kinase, and phosphoglycerate mutase from pig liver and muscle. PMID- 6296622 TI - Galactose-1-phosphate uridylyltransferase from Entamoeba histolytica. PMID- 6296623 TI - D-Tagatose-6-phosphate kinase from Staphylococcus aureus. PMID- 6296624 TI - Inorganic pyrophosphate: D-fructose-6-phosphate 1-phosphotransferase from mung bean. PMID- 6296625 TI - 6-Phosphofructokinase (pyrophosphate) from Entamoeba histolytica. PMID- 6296626 TI - Cloning of the regulatory locus ompB of Salmonella typhimurium LT-2. I. Isolation of the ompR gene and identification of its gene product. AB - We have cloned the complete functional ompB locus of Salmonella typhimurium LT-2 into Escherichia coli K-12 using a cosmid vector and in vitro packaging into lambda. The ompB locus of Salmonella was found to complement both envZ and ompR mutations in E. coli as well as an ompR mutation of Salmonella. The ompR part of the ompB locus was further subcloned into the multicopy plasmid pKN410 as a 1.3 kb fragment. This fragment coded for a single 28.5 kd protein corresponding to about 820 bp in length. Furthermore, the OmpR proteins of S. typhimurium and E. coli were shown to be structurally and functionally highly similar. PMID- 6296627 TI - Cloning of the regulatory locus ompB of Salmonella typhimurium LT-2. II. Identification of the envZ gene product, a protein involved in the expression of the porin proteins. AB - We have isolated the envZ gene from a recombinant plasmid carrying the complete ompB region of Salmonella typhimurium (Liljestrom et al. 1982). The gene was found to be situated very near the ompR gene in the ompB locus. Using minicells, the envZ gene product was identified as a 46 kd protein. Strains harboring either an ompR+ plasmid or an envZ+ ompR+ plasmid were found to differ in their response to environmental changes with respect to porin protein expression. The results obtained suggest that the EnvZ protein, together with ompR, ultimately determines the level of expression of the porin proteins. PMID- 6296628 TI - Interspecific plasmid transfer between Streptococcus pneumoniae and Bacillus subtilis. PMID- 6296629 TI - Cloning and expression of Bacillus subtilis spore genes. PMID- 6296630 TI - Genetic rearrangements in plasmids specifying total degradation of chlorinated benzoic acids. PMID- 6296631 TI - The linkage mapping of cloned restriction fragment length differences in Caenorabditis elegans. PMID- 6296632 TI - Interactions of Tn7 and temperate phage F116L of Pseudomonas aeruginosa. AB - Tn7 insertions into the genome of F116L, a Pseudomonas aeruginosa generalized transducing phage, were isolated by repeated cycles of transduction and induction isolated by repeated cycles of transduction and induction of strains lysogenic for F116cts mutants with selection for trimethoprim resistance (Tpr). Two non defective F116Lcts:: Tn7 phage were characterized. They have reduced plaquing ability, produced non-lysogenic Tpr transductants, and have yielded a deletion mutant of the phage genome upon selection for plaque formation in single infection. F116L DNA is circularly permuted and terminally redundant. A circular restriction map of 61.7 kb has been defined, and a cleavage site common to many enzymes has been identified at coordinate 23.3 kb on the map. It is presumed that this site represents the sequence for the initiation of DNA encapsidation by a headful packaging mode. The Tn7 insertion targets and a 13.4 kb deletion define regions of the F116L genome non-essential for either vegetative growth or lysogenization. The restriction map of Tn7 has been determined for five enzymes. Non-lysogenic Tpr transductants reveal a Tn7 insertion hot-spot in the P. aeruginosa genome. PMID- 6296633 TI - Inverted duplication in the genome of the temperate Streptomyces phage SH3. AB - The DNA of the temperate Streptomyces phage SH3 contains 100 base-pair long inverted repeats separated by a 940 base-pair long segment of DNA as revealed by electronmicroscopic analysis of snapback structures formed after rapid intrastrand reannealing of denatured DNA. The inverted repeat structure was found preferentially at map unit 22 of the circular physical map, in rare cases also in other positions, suggesting a movable character of this genetic element. PMID- 6296634 TI - Maintenance of the bacteriocinogenic plasmid Clo DF13 in Escherichia coli cells. II. Specific recombination functions involved in plasmid maintenance. PMID- 6296635 TI - Effects of Tn5 insertion in the incD region on mini-F maintenance and polypeptide synthesis. AB - The gene coding for the 33,000 dalton protein (B protein) of the mini-F plasmid has been mapped by minicell polypeptide analysis to 47.3-48.2 Fkb. Transcription of the gene is initiated near 47.3 Fkb. Gene B mutants overproduce the 42,000 dalton protein (A protein) of mini-F. Insertions of Tn5 in the B gene and in the incD region cause instability of plasmid inheritance. Mini-F plasmids deleted for part of gene B are not maintained in the absence of the incD region. B protein and the incD region appear to interact in promoting mini-F maintenance. PMID- 6296636 TI - A follow-up study on bovine rotavirus dissemination among calves of a large dairy herd. AB - Rotaviruses had previously been involved in lethal diarrhoea of newborn calves as well as non-lethal diarrhoea in calves aged 6 to 7 weeks in a closed dairy herd comprising 220 heads of cattle. Fifty calves born consecutively were thereafter tested at weekly intervals till 8 weeks old for diarrhoea by clinical judgement of their feces and for rotavirus excretion by ELISA tests. Serum antibodies against rotaviruses were sporadically determined by CF tests. Rotavirus was shown to persist in this herd in widely disseminated form, in seasonal waves, peaking during the second week of age. The first week of life and the third, in this order, ranked next in frequency of dissemination. Silent, non-clinical rotavirus excretion also occurred quite frequently in calves 1 to 3 weeks old. All calves examined possessed maternally-derived antibodies in their blood serum, which according to literature merely exert protective activity as long as present in the unabsorbed state in the gut lumen. Careful colostrum feeding during the first week postponed rotavirus excretion as compared to the earlier onset described in the United States, and milk replacer (devoid of antibody) fed from day 8 onward accounted for the peak of rotavirus infection in the second week of life. It proved impossible to assess the degree of rotavirus coetiology for diarrhoea observed at high frequency. PMID- 6296637 TI - Ultrastructural observations of polyoma infected Friend erythroleukemic cells. AB - Friend erythroleukemic cells (FLC) infected by Polyoma (Py) virus were studied at the electron microscope both during their lytic cycle, which occurs within the first 96 hr after infection, and in the surviving stabilized cells 30 days after infection. The results showed that the Polyoma virus and Friend leukemic virus (FLV) coexist and mature together in the same cell. During the lytic cycle, the Py particles were scattered in the nucleoplasm, and sometimes they were also found aggregated into crystals. Instead crystalline aggregates of Py particles were always present in the few surviving virus-carrying cells 30 days after infection. A possible interpretation could be that the crystal aggregation serves to maintain the viruses in a latency that allows the survival of the host cells. PMID- 6296638 TI - Role of cytoskeleton in natural cell-mediated cytotoxicity against Trichomonas vaginalis. PMID- 6296639 TI - Effect of lead acetate on superoxide anion generation and its scavengers in mice given endotoxin. AB - The administration of endotoxin to mice rendered hypersensitive by lead acetate resulted in profound lipid peroxide formation in the liver 6 hr postintoxication. Endotoxin plus lead acetate administration depressed glutathione peroxidase and superoxide dismutase activities in mouse liver, whereas superoxide anion generation significantly increased in the livers of endotoxin plus lead acetate treated mice compared with that in mice treated with endotoxin alone. Serum acid phosphatase and lactate dehydrogenase isozyme exhibited much more leakage in endotoxin plus lead acetate-injected mice than in sera of mice given endotoxin alone. Nonprotein SH level in the liver was reduced markedly in endotoxin-lead treated mice compared with those receiving endotoxin alone. The plasma vitamin E level was found to decline by 6 hr postintoxication in both endotoxin-lead and endotoxin alone-treated mice, and the transient elevation of the plasma level at 18 hr may be considered to indicate mobilization from other tissues into the blood. PMID- 6296640 TI - Integration of bovine leukemia virus DNA in the genomes of bovine lymphosarcoma cells. AB - Integration of bovine leukemia virus (BLV) in the genomes of infected cells was investigated in cattle with enzootic bovine leukosis (EBL) and sporadic bovine leukosis (SBL). Southern blot hybridization of BLV cDNA to Eco RI and Xba I restriction fragments of EBL tumor DNAs revealed that: 1) one to four or more copies of proviral DNA were integrated per genome; 2) the restriction pattern of the integrated proviral DNA was the same in two or three different tumors from the same animals; and 3) different patterns were observed among tumors from four different animals. These findings suggest the monoclonal origin of different tumors in an individual animal and the existence of multiple chromosomal integration sites of BLV provirus. DNAs from several SBL tumors were also analyzed with the same restriction enzymes, but with both representative and cDNA3'-enriched's of BLV RNA. No hybridization bands reactive with representative BLV cDNA could be detected, while several bands appeared to hybridize with cDNA3' enriched. PMID- 6296641 TI - Neutralization of sendai virus by the IgG subclass antibodies of the guinea pig. AB - A comparative study was made of the neutralizing activities of IgG subclasses IgG1 and IgG2, fractionated from guinea pig antisera against Sendai virus. The yields of IgG2 from the antisera were about 16 times as much as those of IgG1. The neutralizing activity of IgG2 per unit weight was four times as high as that of IgG1. This neutralizing activity of both IgG subclasses was enhanced about 10 times by addition of antibodies to the L-chain of guinea pig immunoglobulin. It is suggested that, in the complement-dependent neutralization of the virus, IgG1 and IgG2 activate the complement through the alternative and the classical pathway, respectively. PMID- 6296642 TI - [Determination of the carbamate kinase activity of bacteria]. AB - The activity of carbamate kinase (EC 2.5.2.2) was determined in bacteria using a simple modified procedure. Carbamoyl phosphate produced under the action of carbamate kinase carbamoylated ammonia in a reaction which was not enzyme catalyzed yielding urea that was assayed by the colorimetric technique. The activity of carbamate kinase was found by this method in a number of microorganisms. The method can be used to study other enzymes synthesizing carbamoyl phosphate. The advantages of the method over other techniques are discussed. PMID- 6296643 TI - The pathogenesis of idiopathic hypercalciuria: an alternative hypothesis. AB - Currently, four hypotheses attempt to explain the pathogenesis of idiopathic hypercalciuria. Proposed theories include hypersecretion of parathyroid hormone, hyperabsorption of calcium by the gastrointestinal tract, hyperreabsorption of calcium by the kidney, and hyperexcretion of phosphate by the kidney. These hypotheses are more descriptive than explanatory. A new hypothesis is suggested which encompasses the divergent findings. Idiopathic hypercalciuria may result from hypersecretion of parathyroid hormone which is active at the gastrointestinal and bone target site but is partially defective at its renal site of action. PMID- 6296644 TI - Neocortex: a high speed analogic conditionable complex-pattern recognizer. PMID- 6296645 TI - Biochemical and biophysical approaches to improving the anticancer effectiveness of Ara-adenine. AB - Ara-C at very low dosage has been reported to decrease the host toxicity of ara AMP or ara-A in combination with 2'-deoxycoformycin, a potent adenosine deaminase inhibitor, while increasing the toxicity to intracerebral L1210 leukemia. The possibility of increasing the selectivity of ara-A by prior administration of ara C is explored. The importance of deoxynucleoside kinases, some of which may be cancer-induced, in obtaining selective anticancer effects is discussed. The possibility of a conformational basis for the differing degrees of selectivity and activity of various novel arabinosyl nucleosides is evaluated. The levels of cyclic nucleotides, which have opposing effects on leukemia, may possibly be manipulated to interfere with the growth of cancer cells. Approaches to minimizing major metabolic distortions, such as the progressive accumulation of dATP associated with the use of potent adenosine deaminase inhibitors and which limit the therapeutic effects of ara-A, are proposed. PMID- 6296646 TI - Coxsackie B virus infection in acute myocardial infarction and adult heart disease. AB - Over a 12-month period, 329 patients admitted to the Coronary Care Unit, Wellington Hospital, New Zealand, were studied for evidence of Coxsackie B virus infection. Fifteen patients (9.8%) with acute myocardial infarction (AMI), three (6.5%) with arrhythmia, and three (25%) with heart failure had serological evidence of Coxsackie B virus infection. During the same period, two control groups were also studied, and the rates of Coxsackie B infection in these groups were compared with that of patients with AMI. The standardised morbidity ratio (SMR) of Coxsackie B infection was 96 for patients with AMI compared with 104.5 for the first control group--that of patients with miscellaneous diseases other than cardiac. The difference between these groups was not significant (P greater than 0.1). However, the SMR of 91.3 for patients with AMI was very high compared with that of 0.0 for the second control group, which consisted of healthy blood donors. PMID- 6296647 TI - [Opiates: effects in animal experiments and in man]. PMID- 6296648 TI - Automated range compensation for proton therapy. AB - A system has been developed to produce complex three-dimensional Lucite compensators to adjust proton depth of penetration within treatment fields. Range adjustments are based on depth prescription contours laid on serial CT scans which contain detailed information on patient contours and inhomogeneities traversed by the beam. The automated system for range compensator production is described, with the constraints arising from proton scattering, fabrication time, and required resolution. Agreement between desired and obtained dose distributions is shown for a typical treatment site. The system's use for automated production of contoured patient apertures is also discussed. PMID- 6296649 TI - [Clinical use of bumetanide in infants with decompensated heart disease]. PMID- 6296650 TI - Initiation of in vitro lipid peroxidation by N-hydroxynorcocaine and norcocaine nitroxide. AB - Norcocaine nitroxide and N-hydroxynorcocaine were found to stimulate hepatic microsomal lipid peroxidation in vitro, as measured by spin-trapping techniques using the spin trap alpha-[4-pyridyl-1-oxide]-N-tert-butylnitrone. It was determined that either norcocaine nitroxide or N-hydroxynorcocaine markedly enhanced the rate of spin trapping of lipid peroxyl radicals when added to hepatic microsomal preparations. Glutathione, in the presence of dialyzed cytosol, inhibited the formation of lipid peroxyl spin-trapped adducts. This finding suggests that cytosolic glutathione-dependent enzymes perhaps including glutathione peroxidase play an important role in the prevention of norcocaine nitroxide-or N-hydroxynorcocaine-mediated lipid peroxidation. PMID- 6296651 TI - Characterization of alpha-adrenergic receptor subtypes in the rat renal cortex. Differential regulation of alpha 1- and alpha 2-adrenergic receptors by guanyl nucleotides and Na. AB - We have used the subtype-selective radioligands [3H]prazosin (an alpha 1 adrenergic antagonist) and [3H]yohimbine (an alpha 2-adrenergic antagonist) to examine alpha-adrenergic receptors in rat renal cortical membranes. Under the conditions used in this study, [3H]prazosin bound only to alpha 1-adrenergic receptors, whereas [3H]yohimbine bound only to alpha 2-adrenergic receptors; the two radioligands were completely selective and did not bind to a common site. The ratio of alpha 2- to alpha 1-adrenergic receptors was about 3:1. Guanyl nucleotides decreased the affinity of epinephrine at both receptor subtypes, but this effect was greater at the alpha 2-receptor and, according to computer analysis, occurred through different mechanisms at the two receptor subtypes. NaCl decreased the affinity of epinephrine at both alpha-receptor subtypes; this effect was more Na+-selective at alpha 2- than at alpha 1-receptors. Guanyl nucleotides and NaCl were additive in decreasing the affinity of epinephrine at the alpha 1-receptor but were synergistic at the alpha 2-receptor. In addition, NaCl increased specific binding of [3H] yohimbine but had no effect on the binding of [3H]prazosin. This enhancement of [3H] yohimbine binding was Na+ specific and fully reversible, and represented an increase in maximal binding capacity. Although binding of epinephrine to both alpha 1- and alpha 2-receptors could be modulated by guanyl nucleotides, we were unable to detect inhibition by epinephrine of basal or hormone-stimulated adenylate cyclase activity. Thus, separate alpha 1- and alpha 2-adrenergic receptors can be detected in the rat renal cortex and binding to both receptor subtypes can be regulated by guanyl nucleotides and Na+ X Na+ may directly interact with alpha 2- but not alpha 1 adrenergic receptors in the renal cortex. Our findings with renal cortical membranes indicate that regulation of agonist binding at alpha-adrenergic receptor subtypes by guanyl nucleotides and Na+ is not limited to alpha 2 adrenergic receptors, as previously reported with other tissues. PMID- 6296652 TI - Parallel observation of the occupancy of the alpha 2-adrenergic receptor in intact platelets and its ability to inhibit the adenylate cyclase. AB - The binding of the selective alpha2-adrenergic receptor antagonist [methyl 3H]yohimbine to intact human blood platelets was studied in parallel with an assessment of the ability of epinephrine to inhibit their accumulation of cyclic AMP in response to prostaglandin E1 and a phosphodiesterase inhibitor. Specifically bound [3H]yohimbine dissociated in a monoexponential fashion, and equilibrium binding showed 200-400 sites/platelet with a dissociation constant of 2-7 nM. The concentration of epinephrine which inhibited cyclic AMP accumulation was one-quarter of that required for an equal degree of inhibition of [3H]yohimbine binding. Conversely, the concentration of yohimbine required to inhibit the action of epinephrine on the adenylate cyclase was 10 times higher than that required for an equal degree of saturation of the alpha2 adrenoreceptor. The ability of epinephrine and the partial agonist p aminoclonidine to compete with [3H]yohimbine for binding was not influenced by agonist-occupation of the ADP receptor, which also inhibits the adenylate cyclase. These results are not compatible with the concept that each adenylate cyclase unit is coupled to an alpha 2-receptor, nor do they indicate that the agonist-occupied alpha 2-receptor forms a complex with the enzyme which persists for the duration of the inhibitory effect. However, it is suggested that these results are compatible with a persistent inhibition of the cyclase being induced by a brief interaction between the agonist-occupied receptor and the adenylate cyclase. PMID- 6296653 TI - Characterization of adenylate cyclase-coupled alpha 2-adrenergic receptors in rat renal cortex using [3H]yohimbine. AB - Alpha 2 adrenergic receptors in rat renal cortex were measured with the antagonist ligand [3H]yohimbine. Renal cortical membranes contained 120 +/- 11.7 (SE) fmoles of binding sites per milligram of protein (n = 9). Their affinity for [3H]yohimbine was 10.4 +/- 0.5 (SE) nM (n = 9) from equilibrium studies and 7.2 +/- 3.3 (n = 4) from kinetic measurements. Alpha-adrenergic agonists and antagonists bound [3H]yohimbine sites with affinities consistent with alpha 2 receptor binding. The slope factors for the binding of all antagonists tested were close to 1, whereas agonists bound with slope factors of less than 1, consistent with the presence of receptors of more than one affinity. The alpha 2 receptors measured with [3H]yohimbine appeared to be coupled to the inhibition of adenylate cyclase. The affinities of alpha-adrenergic catecholamines measured in cyclase inhibition studies were similar to their affinities in binding studies when these were performed under conditions optimal for adenylate cyclase inhibition. Both sodium ion and GTP are required for maximal inhibition of renal cortical adenylate cyclase [Woodcock, E.A., C. I. Johnston, and C.A. Olsson. J. Cyclic Nucleotide Res. 6:261-271, 1980)]. Each of these factors produced a 10- to 20-fold decrease in the binding affinity of epinephrine. Together, a 100- to 200 fold decrease was produced. The concentrations of both sodium ion and GTP required to decrease the affinity of epinephrine were similar to the concentrations required for adenylate cyclase inhibition. This suggested that similar mechanisms were involved in decreasing the affinity of agonists for the alpha 2-receptor and promoting agonist-induced inhibition of adenylate cyclase. PMID- 6296654 TI - Differences in the forskolin activation of adenylate cyclases in wild-type and variant lymphoma cells. AB - The ability of the diterpene forskolin to stimulate cyclic AMP accumulation in intact cell and membrane preparations of wild-type S49 lymphoma cells (WT) and a number of variants has been confirmed. Additionally, a number of salient new findings have emerged: (a) A time delay in forskolin stimulation of cyclic AMP accumulation and adenylate cyclase (t 1/2 approximately equal to 1.5 min) occurred in all hormone-sensitive WT and variant cell and membrane preparations tested. (b) The time delay was missing in the adenylate cyclase-deficient variant (cyc-) of the S49 lymphoma cell, which also lacks functional adenylate cyclase coupling proteins. (c) The simultaneous addition of epinephrine and forskolin to WT cells or to membrane preparations eliminated the time delay. (d) Forskolin stimulation of intact WT cells did not appear to desensitize adenylate cyclase. (e) The activation of WT adenylate cyclase by forskolin was biphasic with respect to concentration, with both high- and low-affinity components being apparent. In cyc-, only the low-affinity component was detected. PMID- 6296656 TI - Meproadifen reaction with the ionic channel of the acetylcholine receptor: potentiation of agonist-induced desensitization at the frog neuromuscular junction. AB - The actions of the nicotinic noncompetitive antagonist meproadifen on both the acetylcholine (ACh) receptor-ion channel complex and electrically excitable membrane were examined in frog sciatic-nerve sartorius muscle preparations. Meproadifen (10-25 microM) blocked the nerve-evoked twitch without affecting the directly evoked twitch, the threshold, overshoot, amplitude, rate of rise, or falling phase of the directly elicited action potential in muscle. This suggests that this agent, at the concentrations that affect the nicotinic receptor, had negligible effect on the excitable membrane. In addition, the drug did not affect either the quantal content or quantal size of the end-plate potential. Meproadifen caused a voltage- and time-dependent decrease in the peak amplitude of the end-plate current (EPC) without significantly shortening the time constant of EPC decay. The voltage- and time-dependent effects of meproadifen were more pronounced at more negative potentials, as evidenced by hysteresis loops and nonlinearity in the current-voltage relationship of the EPC. Both hysteresis and nonlinearity in the current-voltage relationship of the EPC were eliminated when brief conditioning pulses were used for stepwise changes of membrane potentials. The decay time constant of the EPC in the presence of meproadifen remained an exponential function of time. Meproadifen blocked iontophoretically elicited EPCs but did not affect single-channel lifetime, conductance, or the decay time constant of the miniature EPC. Thus, the blockade was more marked on iontophoretically elicited EPCs than on miniature EPCs. Meproadifen also caused desensitization of both the junctional and extrajunctional ACh receptors, but, more important, meproadifen accelerated steady-state desensitization by several fold (compared with the agonist). The marked depression of peak EPC amplitude and miniature EPC, its high affinity for the binding sites in the presence of the agonist, and acceleration of agonist-induced desensitization suggest that meproadifen interacts with the ACh-bound but nonconducting state of the ACh receptor-ion channel complex. Therefore, it appears that meproadifen interacts with the closed ionic channel of the ACh receptor in its resting and activated but nonconducting states, and only slightly affects the open conformation of the ionic channel. PMID- 6296655 TI - Inhibition of binding of [3H]batrachotoxinin A 20-alpha-benzoate to sodium channels by the anticonvulsant drugs diphenylhydantoin and carbamazepine. PMID- 6296657 TI - Etorphine binds to multiple opiate receptors of the caudate nucleus with equal affinity but with different kinetics. AB - The binding of [3H]etorphine, a potent opiate agonist of the oripavine series, to membranes derived from sheep brain caudate nucleus is analyzed. Although the receptors are saturated by [3H]etorphine in a homogeneous fashion with an apparent dissociation constant of 1.16 +/- 0.3 nM, kinetic displacement studies reveal that at least two classes of sites are involved. All of the specific sites for [3H]etorphine are blocked by morphine or naloxone if these ligands are added prior to, or simultaneously with [3H]etorphine. Otherwise, when [3H]etorphine is added prior to morphine or naloxone, only one-third of the specific binding can be effectively displaced. The difference in the displacement patterns between the two classes of sites can be accounted for by the kinetics of the interaction between [3H]etorphine and the receptors. At 37 degrees [3H]etorphine dissociates from one-third of the sites with a half-life of 2.3 min and from the remaining sites with a half-life of 70 min. The sites which release [3H]etorphine slowly have a 10-fold higher apparent affinity for morphine and for naloxone as compared with the more rapidly reversible sites. The binding data are only partially compatible with a model which involves two independent classes of sites. The possibility of identifying the site from which [3H]etorphine dissociates slowly with millimicron, delta, or kappa opiate receptors is explored in light of the fact that [3H]etorphine is a mixture of D- and L-stereoisomers. PMID- 6296658 TI - Structural characterization of beta-endorphin through the design, synthesis, and study of model peptides. PMID- 6296659 TI - Light-enhanced inhibition of ouabain binding to digitalis receptor in rat brain and guinea pig heart by the food dye erythrosine. AB - Erythrosine (ERY) (FD & C red no. 3) inhibited specific binding of [3H]ouabain to rat brain homogenates with an IC50 of 23 microM in the dark and 1 microM in ordinary fluorescent light. Competition studies demonstrated the presence of two components, only one of which was affected by light. Lineweaver-Burk analysis indicated that ERY preferentially antagonizes [3H]ouabain binding at a high affinity site in the light, whereas in the dark the dye inhibits binding in a manner qualitatively similar to inhibition by ouabain. Light enhancement of ERY potency occurred only when dye and tissue were present together in the incubation medium, pointing to participation of transient molecular species. However, neither superoxide dismutase nor catalase altered the effects of ERY in the light or dark, suggesting the absence of oxygen free radicals. When oxygen levels were raised, there was enhancement of inhibition by ERY at a high-affinity receptor accompanied by disappearance of [3H]ouabain binding at one of lower affinity. In contrast to brain, membranes from guinea pig heart showed only one binding site for [3H]ouabain, and antagonism by ERY at this site was markedly enhanced by light. Structural differences between classes of ouabain binding regions probably accounts for the discrimination exhibited by ERY in the presence of light and oxygen. Our findings also caution that metabolic transformation of this common food dye, light decomposition, or photoreaction with foodstuff may yield more toxic derivatives. PMID- 6296660 TI - Formation of ADP-sensitive phosphorylated intermediate in the electric eel Na, K ATPase preparation. AB - The ADP-sensitive and K+ -sensitive phosphorylated forms of Na,K-ATPase (E1P and E2P, respectively) are believed to be the main phosphorylated intermediates of Na,K-ATPase. In the presence of 100 mM Na+, E2P is the major component of the phosphorylated form in all native Na,K-ATPase preparations known, including the microsomes of shark rectal glands. However, the Na,K-ATPase-rich membrane fragments of the electric eel formed a different type of phosphoenzyme, of which 80% was dephosphorylated by both 2 mM K+ and 1 mM ADP within 1 sec at 4 degrees under the same conditions. In the presence of oligomycin (50 micrograms/ml), only the percentage of K+ -sensitive phosphoenzyme was reduced in the eel enzyme, but no such effects were observed in the shark enzyme. However, the eel enzyme produced E2P-rich phosphoenzyme in the presence of 10 mM Na+, as did the shark enzyme in the presence of 100 mM Na+, and the shark enzyme formed the E1P-rich phosphoenzyme in the presence of 500-700 mM Na+, as did the eel enzyme in the presence of 100 mM Na+. These results suggest that the eel enzyme has a much higher affinity for Na+ on the Na+ site controlling the E1P-E2P equilibrium than does the shark enzyme, but we have not been able to explain this difference. Since the phosphorylated forms of both enzymes became more sensitive to both ADP and K+ at 25 degrees, both the interconversion of E1P to E2P and the reverse reaction seem to be accelerated by an increase in temperature. Oligomycin inhibited this conversion of E1P to E2P at 25 degrees as at 4 degrees, but did not inhibit the reverse reaction. Therefore, we conclude that E1P was the predominant phosphorylated form of the eel enzyme under the above conditions, and that E1P was converted to E2P and then dephosphorylated by K+. PMID- 6296661 TI - Interaction between ouabain and the phosphorylated intermediate of Na,K-ATPase. AB - After phosphorylation of electric eel Na,K-ATPase by Na+, Mg2+, and ATP was terminated by removing the unbound Mg2+, the phosphoenzyme was able to bind ouabain upon the addition of 2 mM ouabain under certain conditions. This binding was demonstrated by a 50% inhibition of ATPase after the removal of unbound ouabain by a Sephadex G-50 column (ouabain trapping method). At 4 degrees, this ouabain binding was observed on the K+- sensitive phosphoprotein (E2P) formed in the presence of 10 mM Na+ but was not observed on the ADP-sensitive phosphoprotein (E1P) formed in the presence of 1000 mM Na+. The increase in the dephosphorylation rate of E2P with various concentrations of K+ paralleled the decrease in inhibition by the addition of 2 mM ouabain after the termination of phosphorylation. In 50-200 mM Na+, the eel enzyme used here formed the E1P-rich phosphoprotein, but this phosphoprotein could bind with ouabain, even though the presence of ADP or oligomycin, which prevents the conversion of E1P to E2P, partially interfered with this ouabain binding. At 25 degrees, ouabain binding with E1P-rich phosphoprotein was observed in higher yield (up to 71%), but in each of these cases ADP or oligomycin strongly inhibited ouabain binding. Moreover, ouabain binding with E2P-rich phosphoprotein did not significantly change with temperature, but ouabain binding with E1P-rich phosphoprotein increased more than 6 times at temperatures from 4 degrees to 25 degrees. From these results, it can be concluded that E2P can bind with ouabain in the absence of free Mg2+ whereas E1P cannot, and that the interconversion between E1P and E2P can be stimulated with ouabain binding and accelerated with elevation of temperature. ADP- and K+ -insensitive phosphoprotein probably is only a minor intermediate for ouabain binding. PMID- 6296662 TI - Direct detection of the sulfur trioxide radical anion during the horseradish peroxidase-hydrogen peroxide oxidation of sulfite (aqueous sulfur dioxide). AB - The ESR spectrum of SO3- is observed directly during the oxidation of (bi)sulfite to sulfate by horseradish peroxidase. This radical exhibits a single line at g = 2.0031. The SO3-radical can be trapped with nitrosobenzene, yielding an ESR spectrum with coupling constants AN = 12.3 G,AHp = AHo = 2.4 G, and AHm = 0.9 G, and a g-value of 2.0053. SO3- is an intermediate in the two-step reduction of peroxidase Compound I by (bi)sulfite at physiological pH. At low pH, no SO3- is observed, which indicates a direct, one-step, two-electron reduction of Compound I. The pH at which the mechanism changes depends on the isoenzymes present. The radical reacts rapidly with oxygen as evidenced by the absence of an ESR spectrum when oxygen is present and by oxygen uptake measurements. PMID- 6296663 TI - [Molecular cloning of DNA fragments from Raus sarcoma virus]. PMID- 6296664 TI - [Cloning of bacteriophage T5 DNA fragments in the plasmid pBR322. Analysis of recombinant plasmids by the method of bonding with RNA-polymerase from Escherichia coli on nitrocellulose filters]. AB - DNA of bacteriophage T5 was hydrolyzed with restriction endonucleases HindIII and BamHI, and subjected to the combined hydrolysis with BamHI+EcoRI and BamHI+ +HindIII. Fragments obtained were cloned in the plasmid pBR322. About 17% of T5 genome were recovered in recombinant plasmids. Cloned fragments were localized on the physical map of the phage by restriction analysis and Southern hybridization. With the aim of direct cloning of T5 promoters, PstI/HindIII fragments were inserted into pBR322 followed by selection of recombinants on ApsTCr phenotype. Binding of BsuRI and AluI fragments of hybrid plasmids with E. coli RNA polymerase was studied by nitrocellulose filter assay. The fragments, which were capable to form heparin resistant complexes were identified. PMID- 6296665 TI - [Study by the spin-label method of relaxation properties of protein kinase, its subunits and the catalytic subunit--histone H1 complex]. AB - Using the spin label method, the rotational relaxation in solution of adenosine 3',5'-monophosphate-dependent protein kinase and its subunits as well as the complexes of the enzyme with the substrate, histone H1, was studied. The rotational correlation time of the spin labeled macromolecules was measured on the basis of the quantitative estimation of the label mobility in relation to the protein globule. The holoenzyme molecule was found to be a rigid sphere. Whereas the complex of the globular catalytic subunit of the enzyme with a specific protein substrate, the spin labeled histone H1, appeared a flexible formation. The relaxation properties of the histone H1 molecule selectively labeled by the spin label in its globular part were investigated. PMID- 6296666 TI - [The comparative mapping of virion and cloned DNA of the hepatitis B virus]. AB - The entire hepatitis B virus (HBV) genome and its fragments have been cloned into the BamHI site of the plasmid pBR322 vector. The identity of physical maps of cloned and authentic virion DNAs was demonstrated by restriction enzyme analysis. The location of restriction sites is suggestive of a certain similarity between the studied HBV DNA and HBV DNA, subtype ayw (Galibert et al., 1979). From the restriction enzyme analysis of virion DNA repaired and 32P-labeled by the endogenous DNA-polymerase reaction, the new information concerning the location and maximal length (approximately 1500 nucleotides) of the single-stranded region of HBV DNA has been established. PMID- 6296667 TI - [Fatal granulocyte function defect in a male infant. Results of in vitro and in vivo stimulation with ascorbic acid as well as in vitro stimulation with levamisol and lithium chloride]. AB - We report about a boy who suffered from repeated bacterial infections starting at the age of 4 weeks. A severe defect of chemotactic activity of the neutrophils, and an additional deficient phagocytosis were discovered. The child died at the age of 3 months from septicemia. Chemotactic activity could be stimulated in vitro by ascorbic acid and levamisole but not by lithium chloride. In vivo, however, the effect of ascorbic acid was minimal and treatment with this vitamin could not prevent the lethal end. PMID- 6296669 TI - [Fiber-rich and fiber-poor diets. Use in gastrointestinal diseases]. PMID- 6296668 TI - [Long-term therapy of peptic ulcer disease]. PMID- 6296670 TI - [Dietetics in type II diabetes]. PMID- 6296671 TI - Repair of single-strand breaks in normal and trisomic lymphocytes. PMID- 6296672 TI - Sodium currents in human skeletal muscle fibers. AB - Sodium currents from human external intercostal muscle fibers were recorded using the Hille-Campbell voltage clamp method. Sodium currents are analyzed in terms of the m and h parameters of the Hodgkin-Huxley model. The results indicate that the kinetics and voltage dependence of sodium currents in human skeletal muscle fibers are very similar to those reported in rat fibers. PMID- 6296673 TI - Association of herpesvirus infections with T-lymphocyte-subset alterations, glomerulopathy, and opportunistic infections after renal transplantation. AB - We studied the interrelation among herpes-virus infections, T-lymphocyte subsets, opportunistic infections, and renal histopathology in 28 recipients of renal allografts. All primary or reactivated herpesvirus infections occurring in the first three months after transplantation in recipients of cadaveric grafts accompanied persistent inversions in the ratio of OKT4 (helper/inducer) to OKT8 (cytotoxic/suppressor) lymphocytes. In the less heavily immunosuppressed recipients of organs of living related donors, these inversions were seen only in association with clinically apparent cytomegalovirus infections. Five of seven opportunistic infections occurred in patients with OKT4/OKT8 ratios of less than 1.0. Biopsy specimens from patients with renal dysfunction occurring in association with a low OKT4/OKT8 ratio frequently revealed glomerular damage rather than acute cellular rejection. Monitoring of T-lymphocyte subsets provides early evidence of herpesvirus infections and identifies patients at increased risk for opportunistic infection after renal transplantation. PMID- 6296674 TI - Beta-endorphin and the endocrine pancreas. Studies in healthy and diabetic human beings. AB - Beta-endorphin is present in the endocrine pancreas, suggesting that endorphins may have a role in islet-cell function. To evaluate this possibility, we infused synthetic human beta-endorphin intravenously in healthy volunteers and in insulin dependent diabetic patients. In both groups, beta-endorphin increased plasma glucagon concentrations, and this rise was accompanied by a significant increase in plasma glucose concentrations. In nondiabetic subjects, beta-endorphin also increased plasma insulin concentrations. The threshold dose of beta-endorphin for producing increased plasma concentrations of glucose and glucagon was 0.005 mg--a dose that acutely increased plasma concentrations of beta-endorphin by approximately 40-fold. Glucose, glucagon, and insulin responses to beta-endorphin could not be blocked by intravenous naloxone. These studies suggest that endorphins may be involved in gluco-regulation, that their hyperglycemic action is mediated at least in part by glucagon, and that the effect of beta-endorphin on islet-cell function is relatively resistant to naloxone. PMID- 6296675 TI - Effects of digoxin in infants with congested circulatory state due to a ventricular septal defect. AB - Digoxin alone was used to treat a congested circulatory state in 21 infants (mean age, 2.7 months; mean weight, 3.8 kg) with a ventricular septal defect. The dose was adjusted on the basis of pharmacokinetics to achieve a mean steady-state concentration of 1.6 +/- 0.3 ng of digoxin per milliliter of serum. The mean red cell level of sodium-potassium ATPase fell from 23.1 +/- 7.0 to 12.6 +/- 5.2 nmol per milligram per minute with treatment. Only 6 of the 21 patients had an inotropic response, as reflected by echocardiographic measurements, but the drug was of clinical benefit to 12 infants (including these 6). These results show that not all infants with a congested circulatory state due to a ventricular septal defect benefit from digoxin therapy. Furthermore, in some subjects clinical improvement occurs in the absence of a measurable inotropic response. PMID- 6296676 TI - Peripartum cardiomyopathy. PMID- 6296677 TI - Cytomegalovirus infection complicated by neuralgic amyotrophy. PMID- 6296678 TI - Cushing's syndrome due to ectopic ACTH production by a carcinoid tumor not producing serotonin. PMID- 6296679 TI - No increase in second tumors after cytotoxic chemotherapy for gestational trophoblastic tumors. AB - We investigated the incidence of second tumors after cytotoxic chemotherapy in 457 long-term survivors treated for choriocarcinoma or an invasive mole between 1958 and 1978. Treatment was given according to regular intermittent schedules and over a mean period of four months, with no maintenance. Methotrexate was given to all but two patients, and 261 (57 per cent) also received other cytotoxic drugs, most commonly dactinomycin, vincristine, cyclophosphamide, mercaptopurine, and 6-azauridine. After a mean period of 7.8 years since the beginning of treatment and a total of 3522 patient-years of risk, second neoplasms had developed in only two women (acute leukemia in one and carcinoma of the breast in the other). This figure is less than the number of cases of cancer that would have been expected (3.5) in this group and suggests that the use of methotrexate as chemotherapy for choriocarcinoma is not carcinogenic in the medium term. PMID- 6296680 TI - Renin-secreting pancreatic carcinoma. PMID- 6296681 TI - Delayed de novo methylation in teratocarcinoma suggests additional tissue specific mechanisms for controlling gene expression. AB - Retrovirus infection of embryonal carcinoma cells is blocked at the level of provirus transcription. De novo methylation of the input provirus occurs in embryonal carcinoma cells but not in permissive, differentiated teratocarcinoma. The kinetics of proviral methylation in embryonal carcinoma cells, however, suggest that while methylation may have an important maintenance role in controlling gene expression, additional mechanisms are used by the early embryo to initiate negative gene expression. PMID- 6296682 TI - Unusual splice sites revealed by mutagenic inactivation of an authentic splice site of the rabbit beta-globin gene. AB - Only one of six point mutations of the sequence around one end of the larger of the introns of the rabbit beta-globin gene seriously affects the normal removal of the intron and splicing of the gene. That mutation converts a GT sequence, invariably found at the 5' end of introns, into an AT, which is no longer recognized as a signal for intron removal. Instead, three normally unused (cryptic) sites are used, leading to aberrant gene transcripts. One of the cryptic sites is an exception to the invariable GT sequence. PMID- 6296683 TI - beta-Endorphin regulates lordosis in female rats by modulating LH-RH release. AB - Several lines of evidence have implicated the endogenous opioid peptides in the regulation of masculine sexual behaviour. However, although the opioid related peptides alpha-melanocyte stimulating hormone (alpha-MSH) and adrenocorticotropin (ACTH) have been shown to affect lordosis behaviour in the female rat, there is as yet no evidence for a role of the endogenous opiates in the regulation of female sexual behaviour. We present here evidence that the endogenous opiates in the mesencephalic central grey (MCG) are involved in the control of lordosis behaviour in the female rat. PMID- 6296684 TI - Anxiogenic and non-anxiogenic benzodiazepine antagonists. AB - Benzodiazepines are widely used anxiolytic and anticonvulsant drugs, and brain receptors for these drugs have been characterized by Mohler and Okada and Squires and Braestrup. Recently, substances that antagonize benzodiazepine binding to brain receptors have been discovered. These benzodiazepine antagonists were shown to block the central effects of benzodiazepines and particularly their anticonvulsive properties. Two such antagonists, Ro 15-1788 (an imidazodiazepine) and methyl beta-carboline-3-carboxylate (beta-CCM), have recently been shown to have different intrinsic pharmacological properties. beta-CCM, injected into baboons, cats, mice and rats, is a convulsant, whereas Ro 15-1788 lacks such an activity. Thus, the separation of convulsant and non-convulsant antagonists has been proposed. We suggest here that a subclassification of antagonists is also valid at the behavioural level, based on a conflict model in mice. We show that Ro 15-1788 and beta-CCM antagonize the anxiolytic effect of benzodiazepines. In addition, we find that, when injected alone, Ro 15-1788 has no anxiogenic effects while beta-CCM has anxiogenic properties. We therefore propose that beta-CCM is an anxiogenic convulsant benzodiazepine antagonist and that Ro 15-1788 is a non anxiogenic non-convulsant benzodiazepine antagonist. PMID- 6296685 TI - A dopamine- and cyclic AMP-regulated phosphoprotein enriched in dopamine innervated brain regions. AB - Several mammalian neurotransmitter candidates, for example, serotonin, dopamine and noradrenaline, may exert some of their synaptic effects by regulating protein phosphorylation systems. Comparison of the regional distribution of brain phosphoproteins with neurotransmitter systems may help to identify the specific phosphoproteins involved in the functions of particular neurotransmitters. Here we report the association of one such phosphoprotein with the dopamine pathways in brain. This protein, of apparent molecular weight (MW) 32,000 (32K), seems to be present only in nervous tissue. Its regional distribution within the brain is very similar to the pattern of dopamine-containing nerve terminals; more specifically, the protein appears to be enriched in those dopaminoceptive neurones which possess D-1 receptors (dopamine receptors coupled to adenylate cyclase). The state of phosphorylation of the protein in these dopaminoceptive neurones can be regulated by both dopamine and cyclic AMP. These results suggest that the phosphoprotein may mediate certain of the trans-synaptic effects of dopamine acting on dopaminoceptive neurones. PMID- 6296686 TI - Human class II major histocompatibility antigen beta-chains are derived from at least three loci. AB - Class II antigens of the major histocompatibility complex (MHC) consist of two glycosylated, membrane-integrated polypeptide chains. These cell surface expressed molecules are involved in several immunobiological events involving cell-cell interactions, most of which seem to require that genetically identical class II antigens, or other molecules controlled by the same region of the MHC, are expressed on the interacting cells. The extensive genetic polymorphism of the class II antigens has rendered analyses in the human system of the number of non allelic species of class II antigens difficult, although several laboratories have reported the existence of at least two types of human class II antigens. Here we present the results of experiments using restriction enzyme digestions and separation of DNA from individuals homozygous for the MHC followed by hybridization to human class II antigen alpha- and beta-chain cDNA probes. While the alpha-chain probe gave only a single hybridization band, the various beta chain probes revealed a more complex pattern that is consistent with the existence of at least three separate beta-chain genes or pseudogenes in the human MHC. PMID- 6296687 TI - A mouse type 2 Alu sequence (M2) is mobile in the genome. AB - The Alu and its equivalent families of interspersed repetitive DNA sequences have been found in various mammalian genomes. It has been proposed that some of them might move around the genome like known prokaryotic and eukaryotic transposable elements, as most of these sequences are flanked by short direct repeats at both ends. To prove that this is the case, however, one must demonstrate the existence of homologous sequences of DNA with and without Alu insertion among the genomes of different strains or individuals of a species. While studying a polymorphic repetitive sequence (PR1) originally found in the spacer region of mouse ribosomal RNA genes, we have now found that a sequence similar to the CHO type 2 Alu-equivalent element, designated M2, is inserted within a PR1 sequence which is located outside the ribosomal RNA gene and that this M2 segment is flanked by a short direct repeat at both ends. Furthermore, this PR1 segment containing M2 is detected only in the BALB/c strain among the laboratory mice and wild mouse subspecies examined. These facts suggest that the M2 sequence has been inserted into PR1 sequence relatively recently during evolution of mouse strains and support the idea that at least some of the Alu-equivalent families are mobile in the genome. Recently, Grimaldi and Singer reported an African green monkey alpha satellite sequence that was interrupted by an Alu element. PMID- 6296688 TI - Distinct organizations and patterns of expression of early and late histone gene sets in the sea urchin. AB - The set of histone genes that are active late in embryogenesis of the sea urchin (Strongylocentrotus purpuratus) are present in 5-12 copies per genome and, unlike the clustered, tandemly arrayed early histone genes, are dispersed and irregularly arranged. Late H2B gene expression is activated by events accompanying fertilization and its mRNAs a, first detectable by as early as 6 h of development (16 cells) and increase only slightly in amount during the period of rapid cleavage between 6 and 14 h. However, during the short interval between 14 and 16 h, while the amount of early histone mRNA is declining, there is a greater than 15-fold burst in the rate of late H2B mRNA accumulation. PMID- 6296690 TI - Evolutionary divergence of the mRNA transcription initiation mechanism in yeast. AB - The promoters of eukaryotic genes are being increasingly defined through the identification of consensus DNA sequences, by mutational analysis, and by in vitro and in vivo studies of transcription. Whereas the TATA sequence (Goldberg Hogness box) has been largely conserved among protein encoding genes (transcribed by RNA polymerase II) of eukaryotes, there is some evidence that other structural and functional determinants of mRNA transcription are not conserved between species. I report there an in vivo comparative analysis of the transcription initiation systems of the budding yeast Saccharomyces cerevisiae and the fission yeast Schizosaccharmyces pombe (which can both be transformed by identical plasmids). I have found no instance in which a gene is transcribed in the same fashion in both yeasts. Instead, I have found that the in vivo transcription starting points for many different yeast genes are determined by the cell in which it is transcribed rather than its gene structure alone. The evidence also suggests that the divergence of the transcription initiation system may partly involve the mechanism or structure which determines the distance from the TATA consensus sequence to the site of transcription initiation. PMID- 6296689 TI - T-system optical signals associated with inward rectification in skeletal muscle. AB - The resting potential of many excitable cells, including skeletal muscle, cardiac muscle, nerve cell bodies and egg cells, is determined by a resting potassium conductance which shows inward rectification, allowing potassium ions to move more readily inward across the cell membrane than outward. In skeletal muscle, where inward rectification has been extensively studied, a large part of this conductance is located in the T-system membranes. However, to date, only the kinetic and voltage-dependent properties of this conductance have been studied from analyses of the membrane potential or current recorded at the fibre surface. We report here measurements, obtained using a voltage-sensing dye, of potential changes in the T-system membranes associated with the inwardly rectifying K+ current. Our results show that this conductance alters the time course and significantly attenuates the amplitude of the potential change across the tubular membranes. These optical data provide new evidence for the presence of this conductance in the T-system and, when analysed using a radial cable model for the T-system, provide an estimate of the distribution of the inward rectifier conductance over the surface and T-system which is in agreement with estimates obtained by other techniques. PMID- 6296691 TI - Electrostatic orientation during electron transfer between flavodoxin and cytochrome c. AB - Various studies have shown that reaction rates between reversibly binding electron transfer proteins depend strongly on solution ionic strength. These observations suggest that intermolecular electrostatic interactions are important in facilitating the formation of a productive reaction complex. A recently examined system involves the reduction of vertebrate cytochrome c by bacterial flavodoxin. Although this is a nonphysiological reaction, it proceeds with rates typical for natural partners and is similarly inhibited at high ionic strengths. Here we describe computational studies which examine the role of electrostatics in the formation of a putative reaction complex between flavodoxin and cytochrome c. The results suggest that electrostatic interactions preorient the molecules before they make physical contact, facilitating the formation of an optimal reaction complex. PMID- 6296692 TI - Coffee and opiate receptors. Another cup of coffee? PMID- 6296693 TI - Coffee contains potent opiate receptor binding activity. AB - Opiate receptor-active peptide fragments (exorphins) have been identified recently in casein and gluten hydrolysates, and morphine has been found in bovine and human milk. To determine whether similar peptides or alkaloids occur in other foodstuffs, we have screened potential sources using a rat brain homogenate assay to detect opiate receptor activity. We report here that instant coffee powders from a variety of manufacturers compete with tritiated naloxone for binding to opiate receptors in the rat brain membrane preparations, with no significant difference between normal and decaffeinated coffee. The receptor binding activity resembles that seen with opiate antagonists, in that there was no change in the half-maximal effective dose (ED50) in the presence of 100 mM Na+; on bioassay, the activity was similarly shown to be antagonistic and specific for opiate induced inhibition of twitch. Preliminary characterization of the activity reveals that it has a molecular weight (MW) in the range 1,000-3,500, is heat stable, ether-extractable, not modified by enzymatic digestion with papain, and clearly separable from caffeine and morphine on TLC. As its concentration in an average cup of coffee is five times the ED50, these data suggest that drinking coffee may be followed by effects mediated via opiate receptors, as well as effects of caffeine. PMID- 6296694 TI - Caffeine induces a transient inward current in cultured cardiac cells. AB - Electrical excitation of cardiac muscle may sometimes be due to initiation of inward current by the presence of Ca2+ ions at the inner surface of the cell membrane. During digitalis toxicity and other conditions that abnormally augment cellular Ca2+ stores, premature release of Ca2+ from the sarcoplasmic reticulum leads to a transient inward current, which is large enough to initiate premature beats and is accompanied by a transient contractile response. This inward current may be mediated either by electrogenic sodium-calcium exchange or by specific Ca2+-activated cation channels that have recently been characterized in tissue cultures of cardiac myocytes. An obvious question raised by these observations is whether release of the sequestered Ca2+ stores during each normal beat exerts a similar influence on membrane potential. To explore this, chick embryonic myocardial cell aggregates were voltage-clamped during abrupt exposure to caffeine, which is known to release Ca2+ from the sarcoplasmic reticulum. The speed of the perfusion system and the relative absence of diffusion barriers in the tissue-cultured cells allowed the effects of caffeine-induced Ca2+ release to be studied on a time scale comparable to that of a single normal beat. We report here that abrupt exposure of the cells to caffeine produced a transient inward current having similar features to that of digitalis toxicity, and which was both large enough and rapid enough to potentially contribute to the action potential. PMID- 6296695 TI - Evidence that the insulin secretagogue, beta-cell-tropin, is ACTH22-39. AB - The pituitary neurointermediate lobe of genetically obese (ob/ob) mice contains a hormone which stimulates insulin release and which cross-reacts with a -COOH terminal ACTH antiserum, suggesting that it is related to corticotropin-like intermediate lobe peptide (CLIP), the 18-39 fragment of ACTH. The hormone, which we have called beta-cell-tropin, has been shown to be present in the plasma of the ob/ob mouse and to potentiate glucose induced insulin secretion. We have now shown that ACTH22-39 prepared by tryptic digestion of human synthetic CLIP behaves similarly on Biogel chromatography and on reverse-phase HPLC to the naturally occurring beta-cell-tropin. Furthermore, beta-cell-tropin and ACTH22-39 have indistinguishable antigenic and insulin releasing properties. PMID- 6296696 TI - Homology between human bladder carcinoma oncogene product and mitochondrial ATP synthase. AB - More than 10 different dominant transforming genes (oncogenes) have been identified in human tumours. A human bladder carcinoma oncogene, closely related in sequence to retroviral transforming genes, is split into four exons; the first encodes the N-terminal 37 residues of p21, a protein of unknown function. The oncogene is activated by a single point mutation (guanine to thymine) resulting in the change glycine to valine at position 12 of p21 (refs 3, 4). We report here that the amino acid sequence surrounding this residue is highly homologous to the beta-subunit of mitochondrial and bacterial ATP-synthase in the region of the polypeptide that is believed to contribute to nucleotide binding. Thus, p21 may form part of an enzyme that uses purine nucleotides in catalysis. This is consistent with the finding that an equivalent murine oncogene product binds GTP. PMID- 6296697 TI - Uncoupling of initiation site cleavage from subsequent headful cleavages in bacteriophage T1 DNA packaging. AB - The packaging of intracellular DNA into heads is a key feature in the morphogenesis of bacteriophage particles. In many phages a performed empty head precursor, the prohead, is filled with DNA from a concatemeric substrate consisting of tandemly repeated genome lengths. The addition of outer shell proteins completes head formation. The DNA molecules released from particles of the coliphage T1 exist as three major permutations of nucleotide sequence. Such limited permutation can be explained by the modification of Streisinger's 'headful' mechanism proposed for phage P22. DNA packaging is initiated at a specific site (the pac site) on the concatemeric precursor. While this site is cleaved, subsequent cleavages (headful cleavages) are dependent only on head filling and are not defined in terms of nucleotide sequence. Headfuls of DNA, consisting of slightly more than a genome length, are packaged in three successive cycles of head-filling to produce the permuted and terminally redundant molecules characteristic of T1 DNA. To elucidate the regulation of this process, we have studied the DNA metabolism of T1 head mutants. We describe here the properties of a mutant in gene 13.3 which is defective for headful cleavage but remains proficient in pac site cleavage. The observation in this mutant that concatemers are degraded to unit-length molecules by repeated pac site cleavage suggests a model of headful packaging in which pac site initiation and processive head-filling compete for the DNA substrate. PMID- 6296698 TI - Rearranged mitochondrial genes in the yeast nuclear genome. AB - We have found a contiguous DNA sequence in the yeast nuclear genome with extensive homology to non-contiguous yeast mitochondrial DNA sequences. Closely linked to this nuclear sequence in some, but not all, yeast strains is a tandem pair of transposable (Ty) elements. Certain features of the content and organization of this nuclear DNA sequence suggest that it may have originated from petite mitochondrial DNA which integrated into the nuclear genome. PMID- 6296699 TI - Selective stimulation of dopamine and noradrenaline autoreceptors by B-HT 920 and B-HT 933, respectively. PMID- 6296700 TI - Brain levels of tofizopam in the rat and relationship with benzodiazepine receptors. AB - The effect of tofizopam on 3H-flunitrazepam binding was studied in rat hippocampus and cerebellum. Tofizopam (at a concentration of 10(-7) M) increased 3H-Flu binding through a 30% rise in the Bmax with no modification of Kd in either brain area. Similar results were obtained when the binding was measured in tofizopam (50 mg/kg p.o.) pretreated rats. Even though tofizopam has no anticonvulsive action against pentetrazol-induced convulsions, it significantly potentiated the action of diazepam but with no modification of brain diazepam levels and metabolism. The brain levels of tofizopam are reported and compared to plasma levels after oral administration of 5 and 50 mg/kg to rats. PMID- 6296702 TI - Influence of 16 beta formylation on Na, K-ATPase inhibition by cardiac glycosides. AB - The inhibitory effect of formylated cardiac steroids (gitaloxin and its derivatives) on guinea-pig heart Na, K-ATPase was compared to that of other cardiac steroids with various hydroxy substituents. The decreasing order of potency of aglycones at equilibrium was as follows: gitaloxigenin greater than digitoxigenin greater than ouabagenin greater than digoxigenin greater than gitoxigenin greater than diginatigenin. This sequence was different to the sequence of drugs hydrophobic character. The compounds with hydroxy groups in the vicinity of the lactone ring (gitoxigenin, diginatigenin) were less potent than the hydrophilic compound ouabagenin. We propose that intramolecular bounding between 16 beta-OH and the lactone ring contributes to the relatively low potency of gitoxigenin and diginatigenin. The formylation of 16 beta-OH increased the potency of gitoxigenin by a factor of 41. The formylated compound (gitaloxigenin) was 5-fold more potent than digitoxigenin. The 3 beta-glycosylation of digoxigenin lead to pseudo-irreversible inhibitors of Na, K-ATPase. The half-time to achieve the equilibrium (for 5 mumol/l) was equal to 54 s, 90 s and 108 s respectively for digoxigenin monodigitoxoside, digoxin and desacetyllanatoside C. However, at equilibrium the three glycosides were equipotent, suggesting the existence of steric effects at the sugar site of the receptor. The sequence of potency observed for monodigitoxosides, monodigitalosides and tridigitoxosides after 60 min incubation was similar to that observed for the corresponding aglycones. These results suggest that the strongly negative inductive group 16 beta-OCHO is tightly bound to Na, K-ATPase, possibly to the same receptor site than that which is thought forming hydrogen and ionic bonds with the lactone ring. They show that the high toxicity of gitaloxin in guinea-pig heart is likely due to its high potency as Na, K-ATPase inhibitor. PMID- 6296701 TI - Actions, interactions, and apparent affinities of various ceveratrum alkaloids at sodium channels of cultured neuroblastoma and cardiac cells. PMID- 6296704 TI - Evidence supporting the existence of presynaptic alpha-adrenoceptors in the regulation of endogenous noradrenaline release upon hepatic sympathetic nerve stimulation in the dog liver in vivo. AB - The existence and functional significance of presynaptic alpha-adrenoceptors within the liver was investigated in anesthetized dogs. The stimulation-evoked endogenous catecholamine overflow was determined in hepatic venous blood upon hepatic nerve stimulation (12 V, 1-8 Hz, 1 min). Under resting conditions, average plasma catecholamine levels in hepatic venous and aortic blood were 0.064 ng/ml and 0.334 ng/ml, respectively. A frequency-dependent increase (P less than 0.001) was found in the hepatic venous catecholamine overflow, while aortic catecholamine levels did not change significantly at any stimulation frequency tested. The increases in catecholamine overflow were associated with decreases in hepatic arterial vascular conductance (35-66%, P less than 0.001) at all frequencies tested. Yohimbine (0.3 mg/kg, i.v.) potentiated the catecholamine overflow by 110-140% (P less than 0.05) upon stimulation at low frequencies (1-4 Hz). Clonidine (20 micrograms/kg, i.v.) inhibited the catecholamine overflow by 55-76% (P less than 0.05) at frequencies of 1 and 2 Hz, and reduced the hepatic arterial response by 46% (P less than 0.01) at 1 Hz. The pretreatment with yohimbine (0.1 mg/kg, i.v.) abolished the inhibitory effects of clonidine both on the catecholamine overflow and the hepatic arterial responses. The results support the existence of a negative feedback mechanism mediated by presynaptic alpha-adrenoceptors in the local regulation of noradrenaline release from hepatic sympathetic fibers. A functional significance of this process was suggested by an improved correlation found in the presence of clonidine between the catecholamine overflow and the hepatic arterial vascular conductance. PMID- 6296705 TI - [Medicine with a handicap]. PMID- 6296703 TI - Competitive and non-competitive interactions between specific ligands and beta adrenoceptors in living cardiac cells. AB - We have used primary cultures of hearts from newborn rats to study beta adrenoceptor properties in living myocardial cells. Receptors were labelled with the lipophilic antagonists 3H-(+/-)-carazolol and 125I-(+/-)-cyanopindolol (CYP) or with the hydrophilic antagonist 3H-(+/-)-CGP 12177. Under equilibrium conditions all ligands bound to a saturable homogeneous class of specific sites with a maximal binding capacity of approximately 100 fmol/mg protein (corresponding to approximately 5000 sites/cell). After 90-180 min preincubation of intact cells with 3H-carazolol or 3H-CGP 12177 only 80% of these antagonists could be displaced from specific binding sites by competing ligands. In the simultaneous presence of the antagonist (-) timolol 100% of specifically bound radiolabelled ligand remained displaceable. In competitive displacement experiments the radioligands did not affect the apparent affinity of the displacing nonlabelled antagonists timolol and CGP 12177, but agonist affinity was markedly changed. The apparent KD values for (-)-isoprenaline were 1560 and 2720 nmol/l in the presence of carazolol and CYP, but only 32 nmol/l in the presence of CGP 12177. This antagonist-dependent difference in agonist KD values was observed only in intact cells but not in membrane particles prepared from heart homogenates of newborn rats, where high agonist affinity was seen during displacement of all radioligands. The KA value for isoprenaline-stimulated cAMP accumulation in living cells was 30 nmol/l in 5-day cultures. A direct proportionality existed between agonist receptor occupation and cAMP accumulation in the presence of CGP 12177 as estimated by the KA/KD ratio. In the presence of carazolol the KA/KD ratio decreased from 1 to 0.02 suggesting that low affinity receptors were not coupled functionally to adenylate cyclase. These results indicate that some lipophilic antagonists which appear to be inert competitive ligands in fragmented membranes, alter receptor binding properties in intact cells. These antagonists seem to promote the transformation of receptor sites into a new "inactivated" state where competitive interactions between different ligands are inhibited. PMID- 6296706 TI - [Distribution of the terminals of primary afferent fibers polysynaptically connected to motoneurons in the spinal cord in the frog]. AB - In experiments carried out on an isolated perfused frog spinal cord connected with the hindlimb nerves primary afferent fibres and motoneurons were studied by intracellular electrical stimulation and recording with subsequent injection of HRP. It is found that collaterals of fast-conducting fibres belonging to the cutaneous or muscle nerves and establishing polysynaptic connections with motoneurons terminate within the upper and middle portions of the dorsal horn. Terminal branching of these fibres are characterized by numerous collaterals originating at short distances from each other. In some cases HRP-positive interneurons with axons terminating in immediate proximity to injected fibres were found. It is suggested that HRP-positive interneurons make axo-axonal contacts with terminals of primary afferent fibres. PMID- 6296707 TI - [Differences in the blocking action of benzocaine and amino compounds on batrachotoxin-modified node of Ranvier sodium channels]. AB - Voltage-clamp experiments on the frog Ranvier node have shown that batrachotoxin (BTX) decreases drastically the sensitivity of sodium channels to the blocking action of various tertiary (procaine, trimecaine, ajmalin, strychnine) and quaternary (QX-572, N-propylajmaline) amine drugs but does not affect noticeably sodium channels blockade produced by neutral benzocaine. The inhibition of BTX modified sodium current by large concentrations of trimecaine (3.5 mM) is time- and voltage-dependent. Neither of the amines used proved to be able to cause frequency-dependent (cummulative) block of modified channels. Unblocking of these channels during washing of the node with Ringer solution proceeds much faster than that of normal channels. On the contrary, restoration of normal and modified currents blocked previously by benzocaine has the same time course. The relationships between "receptors" for BTX and various sodium channel blockers are discussed. PMID- 6296708 TI - Carcinoma at the hilus of the liver: clinical, radiological, histological and therapeutic aspects. PMID- 6296709 TI - Neurologic disorders in patients with renal failure. PMID- 6296710 TI - A simple disc-washing apparatus for nucleoside kinase assays. PMID- 6296711 TI - [Sibling case of osteosclerosis with cranial nerve symptoms]. AB - We experienced two cases of "Osteosclerosis" who were 12 and 15 year old sisters. Previous reported cases of this disease are 50 cases and among them only one patient was reported in Japan. Osteosclerosis seems to be inherited as an autosomal recessive trait. Parental consanguinity is also observed. The peculiar facies are evident in infancy, characterized by broad, flat nasal bridge, ocular hypertelorism and prognathic, broadened mandible. Commonly, they have cutaneous syndactylies in bilateral hands and feet, especially between the second and third finger and toe. Roentgenographically, hyperostosis with osteosclerosis can be observed in systemic bones, particularly the calvarium is greatly thickened. Since such a bony change occurs most severely at the base of the skull, important clinical symptoms of this disease are cranial nerve palsies resulting from obliterations of unilateral or bilateral several cranial nerve foramina. In many cases deafness due to progressive encroachment upon the middle ear cavities and auditory nerve canals appears early in infancy. Transient palsy of the facial nerve occurs somewhat later, and bilateral facial paralyses are usually permanent in adulthood. In some cases optic atrophy and visual field defect due to compression of the optic nerves are late complications. Other ocular symptoms are strabismus, nystagmus and exophthalmos. Anosmia and trigeminal nerve palsy are less common. Lower cranial nerve symptoms can not be noted but the reason is unclear. Chronic headache, convulsion and mental retardation are occasionally present. They are considered as a result from increased intracranial pressure due to progressive diminution of the cranial capacity. By same mechanism, several patients have died suddenly from impaction of the medulla oblongata in the foramen magnum in early adulthood. Then, some reporter puts emphasis on prophylactic opening of the foramen magnum in all adult cases. PMID- 6296712 TI - Lesions of the ventral noradrenergic (norepinephrinergic) bundle affect brain and pituitary pools of endorphins and the response of these to stress in rats. AB - The present study examines the interrelationship of the ventral noradrenergic (norepinephrinergic) bundle (VB) with brain and hypophyseal pools of endorphins. As compared to sham-operated rats, selective destruction of the VB depressed hypothalamic levels of norepinephrine and abolished the fall in these levels evoked by acute foot shock stress. Stress elevated plasma levels of beta endorphin immunoreactivity (beta-EI) and both basal and poststress circulating levels of beta-EI were significantly greater in lesioned as compared to sham animals on days 4 and 10, but not 28, postsurgery. At each time, stress depleted the anterior and neurointermediate lobe content of beta-EI in both sham-operated and lesioned rats. These data indicate that stress mobilizes beta-EI from the pituitary into plasma and that the VB may inhibit the tonic and stress-elicited secretion of beta-EI into the circulation. In lesioned rats, a transient depression in basal levels of beta-EI in the septum and in those of met enkephalin immunoreactivity (M-EI) in periventricular midbrain tissue was seen. Stress selectively depleted the beta-EI as compared to the M-EI content of discrete brain regions, including the septum and this periventricular tissue, in sham-operated animals. Lesioned rats also responded to stress with a fall in beta EI levels and, in contrast to sham rats, with a decrease in the M-EI levels of the hypothalamus and periventricular tissue. These data demonstrate that the VB does not mediate the activation of brain beta-EI elicited by stress and suggest the existence of an interaction of the VB with certain brain pools of beta-EI and M-EI both tonically and under stress. PMID- 6296713 TI - Familial nemaline myopathy. AB - Two sisters with congenital nemaline myopathy are described. In both cases almost 70% of muscle fibers contained rods which were selectively localized in the larger ones. The variability coefficient was abnormally increased. Histochemical reactions showed that almost all the muscle fibers were type 1. In one case many fibers contained one or more core-like lesions. The parents and two siblings of the patients were clinically normal; EMG examination also showed normal motor unit potentials. Muscle biopsy was normal in the father; in the mother a slight type 1 predominance was detected without rods or other signs of myopathy. The disease seems to be transmitted by an autosomal recessive trait, although incomplete penetrance of a dominant trait cannot be excluded. PMID- 6296714 TI - Lack of structural abnormalities in lymphocytes from heterozygotes of juvenile type of generalized ceroid-lipofuscinosis. A light and electron microscopic study. PMID- 6296715 TI - Enkephalin increases the efficacy of dopaminergic transmission in Helix pomatia. AB - A dopaminergic synaptic connection in Helix pomatia was studied. Transmitter mobilization was measured during incubation with met-enkephalin. Enkephalin decreased the time constant for transmitter mobilization. The effect was blocked by naloxone. It is postulated that the presynaptic cell possesses opiate receptors and that these receptors mediate the enkephalin-induced increase in efficacy of dopaminergic transmission. PMID- 6296716 TI - Morphine and nigrostriatal function in the rat and mouse: the role of nigral and striatal opiate receptors. AB - Using local injections of drugs and hemisection experiments, a comparison was made of the actions of morphine on the dopaminergic nigrostriatal system of the rat and mouse. In the rat, morphine appeared to act exclusively at presynaptic opiate receptors on dopaminergic nerve endings in the striatum. Activation of these receptors resulted in enhanced dopamine synthesis but with no associated increase in dopamine release. In the mouse, morphine acted at the level of the substantia nigra to enhance both striatal dopamine synthesis and release. The exact localization of these receptors in the substantia nigra remains to be determined. PMID- 6296717 TI - Characterization of solubilized benzodiazepine and muscimol binding sites from rat brain. AB - Binding sites for muscimol and flunitrazepam were solubilized and their properties studied by ultrafiltration, isoelectro-focusing and cation-exchange chromatography methods. The heterogenity of binding sites for flunitrazepam was demonstrated in the ultrafiltration experiments: the binding sites with a molecular weight of more than 300,000 dalton were sensitive to stimulation by gamma-aminobutyric acid (GABA) and those with a molecular weight of less than 300,000 dalton were not. The binding sites for flunitrazepam appeared to be low acidic molecules with pI 5.6-6.0 and binding sites for muscimol an apparently more heterogenous distribution in the range of pH 4.6 to 5.9 was detected. Cation exchange chromatography revealed two subpopulations of binding sites for muscimol one of which copurified together with binding sites for flunitrazepam. Each subpopulation exhibited only one class of binding site with Kd = 20 nM for a subpopulation copurifying together with binding sites for flunitrazepam and Kd = 9 nM for the other. PMID- 6296718 TI - Effects of dithiothreitol, 5,5'-dithiobis(2-nitrobenzoic acid) and N ethylmaleimide on synaptic transmission at sympathetic ganglion cells of frog. AB - The effects of disulfide bond and sulfhydryl group reagents on synaptic transmission were studied in the bullfrog sympathetic ganglion. Ten millimoles of dithiothreitol (DTT), a disulfide bond reducing agent, decreased the amplitude of the transmitted postganglionic compound action potential (CAP) to 41% of control. More than one hour wash-out of DTT with normal Ringer's solution did not reverse the block, but 1 mM 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB), an oxidizing agent, rapidly restored the compound action potential amplitude. N-Ethylmalemide (NEM), an alkylating agent, added before DTNB, prevented the restoration of transmission by DTNB. Block of transmission induced by dithiothreitol was also reversed rapidly by 50 microns 3,4-diaminopyridine (3,4-DAP), to 87% of control compound action potential amplitude, and subsequent addition of DTNB restored transmission completely and this was accompanied by the stimulus-bound repetitive firing (SBR) which diaminopyridine produced in otherwise untreated ganglia. One to 3 mM dithiothreitol decreased the amplitude of excitatory post-synaptic potentials (EPSP), and greater concentrations slightly depolarized the cell membrane and decreased membrane resistance. Washout with Ringer's solution reversed the depolarization, but not the block of excitatory postsynaptic potentials. The amplitude of excitatory post-synaptic potentials was restored to threshold by 1 mM DTNB, which also slightly increased membrane resistance and the resting potential. It is concluded that the actions of dithiothreitol and DTNB in the bullfrog sympathetic ganglion are generally similar to those previously observed in other junctional tissues. Interactions between these drugs and 3,4 diaminopyridine in the present study also raise the question whether dithiothreitol and DTNB have any presynaptic effects. PMID- 6296719 TI - Evidence for multiple "Kappa" binding sites by use of opioid peptides in the guinea-pig lumbo-sacral spinal cord. AB - Binding properties of [3H]-etorphine and [3H]-ethylketocyclazocine have been studied in the lumbo-sacral spinal cord of guinea-pig which does not contain mu or delta binding sites. [3H]-etorphine binds to a single class of high affinity sites, whereas [3H]-ethylketocyclazocine interacts with a high and a low affinity component. Using a discriminative procedure, 5 microM (D-Ala2, D-Leu5) enkephalin (DAL), the high affinity component of [3H]-ethylketocyclazocine can be resolved in two classes of sites, (D-Ala2, D-Leu5) enkephalin sensitive sites (DALS sites) and (D-Ala2, D-Leu5) enkephalin insensitive sites (DALI sites). In these conditions, there is a total loss of [3H]-etorphine sites, whose binding capacity and properties strictly correspond to the DALS sites labelled by [3H] ethylketocyclazocine. Pharmacological investigations indicate that DALI sites for which dynorphin (1 leads to 17) is the best ligand, can be related to kappa sites previously described in guinea-pig brain, whereas DALS sites for which (Arg6, Phe7) Met-enkephalin possesses a good affinity, closely correspond to benzomorphan sites recently characterized in rat brain and spinal cord. [3H] ethylketocyclazocine interacts additionally with "non opiate" low affinity sites, for which only benzomorphan drugs exhibit a good affinity, whereas morphine, naloxone, phencyclidine or endogenous opioid peptides do not present any affinity for them. On the basis of these data, a new subdivision of "kappa" sites is discussed. PMID- 6296720 TI - A serine-proteinase inhibitor (Boc-D-Phe-Pro-Arg-H) inhibits the secretion of adrenocorticotropin- and beta-endorphin-immunoreactive peptides in vitro. AB - The effects of a potent serine protease inhibitor, Boc-D-Phe-Pro-Arg-H, on the secretion and content of ACTH and beta-endorphin-like immunoreactivities of cultured rat anterior pituitary cells were studied. Basal release of the hormones was not affected by the drug but secretion in response to stalk-median eminence extracts was inhibited. The inhibitor did not effect the amount of ACTH or beta endorphin in the medium plus the cells, nor did it change the ratio of the lipotropin and beta-endorphin-like immunoreactivities. Thus the compound seems to affect the release of ACTH and beta-endorphin, but not the processing of lipotropin to beta-endorphin. PMID- 6296721 TI - The development of beta-adrenergic receptors in the visual cortex of the rat. AB - The development of beta-adrenergic receptors in the rat visual cortex was examined and the density of beta-receptors and associated subtypes (beta 1 and beta 2) was compared between visual and non-visual or whole cortical tissues using radioreceptor assays employing [125I]iodohydroxybenzylpindolol and [125I]iodocyanopindolol as ligands. Saturation assays revealed not only similar affinities of beta-receptors for [125I]iodohydroxybenzylpindolol in visual cortical samples at 10, 24 and 160 days after birth but also practically identical saturation curves for visual and non-visual cortical samples at 160 days of age. Displacement of [125I]iodohydroxybenzylpindolol with propranolol in visual cortical membranes at various postnatal ages showed a gradual increase in receptor density from day 4 to day 24 with no change thereafter. No significant differences were observed in the overall density of beta-receptors or in the distribution and density of beta 1 and beta 2-receptors between visual and non visual or whole cortical samples; however, there was a definite decline in the density of beta-receptors in these samples between 40 and 160 days of age. The results indicate that the developmental pattern of beta-receptor density and the distribution of beta 1 and beta 2-receptors are similar between visual and whole cortical tissues. In addition, the results emphasize the importance of maintaining the dissociation constant at a fixed value when comparing receptor densities between experiments, and also show the utility of employing the high affinity ligand, [125I]iodocyanopindolol, with a combination of serotoninergic, dopaminergic and alpha-adrenergic antagonists to examine beta-adrenergic receptors in a specific region of the brain. Study of beta-receptors in the visual cortex may be beneficial in elucidating the role of norepinephrine in this region. PMID- 6296722 TI - Lithium entry into neural cells via sodium channels: a morphometric approach. AB - Rat cerebral cortical explants prepared from 18-day-old embryos were grown for 18 days in vitro. Cultures were exposed to Na+, Li+ and choline+ media, respectively, in the presence or absence of tetrodotoxin and/or veratridine, and processed for electron-microscopy. Veratridine (50 microM) induced an increase in summated and mean areas of neuronal profiles in Na+- and Li+-media but not in the choline+-medium: the summated perimeters did not change. The mean value of the neuronal form factor was significantly elevated following exposure to veratridine in a Na+- or Li+-dependent manner, indicating that the shape of the sectioned neuronal elements shifted towards an (ideal) circle. Qualitative assessment revealed an increased electron-lucency of the cytoplasm of neuronal profiles in Na+- and Li+-media containing veratridine. The veratridine-induced neuronal changes were inhibited by simultaneous addition of tetrodotoxin (1 microM) to the media. In the case of the glial cells, the values of the summated area and form factor did not change in the various experimental groups. The area of the extracellular space per unit area of sections significantly decreased in the Na+- and Li+-media following veratridine exposure; this did not occur in the choline+ medium. The results indicate a considerable swelling of the neuronal elements, reflecting cation, Cl- and water uptake following prolonged sodium channel activation in the presence of Na+ or Li+ ions. The quantitative ultrastructural data strongly suggests an entry of Li+ ions into cultured rat cerebral cells via sodium channels. PMID- 6296723 TI - Evidence for the existence of alpha- and beta-adrenoceptors on cultured glial cells--an electrophysiological study. AB - The action of adrenergic alpha- and beta-agonists and antagonists has been studied on the membrane potential and resistance of glial cells of cultured rat central nervous system. Noradrenaline and the alpha-adrenoceptor stimulating agents phenylephrine and clonidine (10(-7) to 10(-4)M) depolarized the glial membrane, whereas the beta-agonist isoprenaline caused a hyperpolarization at low concentrations (10(-7) and 10(-6)M). The effects of noradrenaline and phenylephrine were reversibly blocked by the alpha-antagonist phentolamine, whereas those of isoprenaline were antagonized by the beta-blocker atenolol. Atenolol did not affect the depolarization by noradrenaline. The glial depolarization induced by the alpha-agonists was not the consequence of a change in the extracellular K+-concentration unlike that produced by amino acid transmitters. The present results, together with those of biochemical and autoradiographic binding studies, suggest that alpha- and beta-adrenoceptors occur on glial cells and that the glial depolarization is mediated by alpha receptors, whereas the hyperpolarization is due to activation of beta-receptors. PMID- 6296724 TI - Evidence for the existence of alpha- and beta-adrenoceptors on neurones and glial cells of cultured rat central nervous system--an autoradiographic study. AB - The cellular localization of the binding of radioactive noradrenaline and alpha- and beta-adrenoceptor antagonists was studied in organotypic cultures of rat cerebellum, brain stem and spinal cord using autoradiography. In cerebellar cultures, many neurones, which appeared to be Purkinje cells, were labelled by [3H]noradrenaline and by the beta-antagonists [3H]dihydroalprenolol and [3H]carazolol, whereas no binding of the alpha-antagonists [3H]prazosin and [3H]rauwolscine was detected. In cultures of spinal cord and brain stem, [3H]noradrenaline and the beta-antagonists were bound to many large neurones. Binding of [3H] alpha-antagonists was observed to a small number of brain stem and spinal neurones, the labelling being much weaker than that produced by the [3H] beta-antagonists. The antidepressant [3H]desmethylimipramine was bound to many neurones and glial cells in cerebellar, brain stem and spinal cord cultures. Glial cells also possessed binding sites for [3H]noradrenaline and alpha- and beta-adrenoceptor antagonists, findings that are consistent with recent electrophysiological observations which indicate the existence of alpha- and beta adrenoceptors on cultured astrocytes. PMID- 6296725 TI - Peripheral neuropathy associated with polycythemia vera. AB - Nerve conduction studies were performed on 26 patients with polycythemia vera, 11 of whom had sensory symptoms in the extremities and 3 of whom had clinical signs of peripheral neuropathy. There was significant impairment of sensory conduction in the ulnar and sural nerves and mild slowing of motor conduction in the lateral popliteal nerve. Sural nerve biopsies were performed on three patients. The pathological findings, including teased fiber studies, were consistent with mild chronic axonal degeneration. There is an association between polycythemia vera and peripheral neuropathy. PMID- 6296726 TI - Monoclonal IgM kappa antibody precipitating with chondroitin sulfate C from patients with axonal polyneuropathy and epidermolysis. AB - We studied two patients with an axonal type of polyneuropathy, epidermolysis, and IgM kappa plasma cell dyscrasia. The IgM kappa was deposited in the dermis, was absorbed from the serum by axonal micelle preparations, and was precipitated with chondroitin sulfate in highly purified agarose in 0.15 M NaCl with 0.01 M phosphate buffer, pH 7.8. In contrast, we found none of these abnormalities in three patients with IgM plasma cell dyscrasia and demyelinating neuropathy. Of 78 other macroglobulinemic serum samples from patients without neuropathy, 7 precipitated with a sulfated polysaccharide. This reaction occurred at low ionic strength, 0.05 M barbital buffer, pH 8.1, but did not occur in the higher ionic strength of 0.01 M phosphate with 0.15 M NaCl (PBS). The interaction of the IgM with chondroitin sulfate at relatively high ionic strength could cause both the axonal polyneuropathy and the epidermolysis. PMID- 6296727 TI - Osteosclerotic myeloma and peripheral neuropathy. AB - Sixteen cases of osteosclerotic myeloma and peripheral neuropathy (SM-PN) were reviewed. The neuropathy resembled chronic inflammatory-demyelinating polyneuropathy with predominantly motor disability, high CSF protein levels, and low motor nerve conduction velocities. Twelve of the 16 patients had detectable levels of monoclonal serum proteins, all with lambda light chains, but results of other laboratory studies were usually normal. Most of the patients also had organomegaly, endocrine abnormalities, or both. Treatment of solitary lesions with tumoricidal irradiation usually improved the neuropathy and reversed the nonneurologic abnormalities; chemotherapy for multiple osteosclerotic lesions was less helpful. PMID- 6296728 TI - Fatal peripheral neurolymphomatosis after remission of histiocytic lymphoma. AB - A 32-year-old woman with histiocytic lymphoma was in complete clinical remission after two courses of chemotherapy, when peripheral neuropathy developed fulminantly. Abnormalities included facial nerve paralysis, dysphagia, quadriparesis, myalgia, and incontinence. She died 10 days after onset of these symptoms. Postmortem examination revealed infiltration of peripheral nerves by lymphomatous cells with no involvement of meninges, brain, lymph nodes, or other organs. Differences in the blood-brain barrier of peripheral and central nervous system are suggested: The peripheral barrier may be more penetrable by malignant histiocytes or less permeable to cytotoxic drugs. Intrathecal chemotherapeutic drug instillation and irradiation may be beneficial. PMID- 6296729 TI - [Screening programs for breast carcinoma]. PMID- 6296730 TI - [Anatomoclinical aspects of ductal carcinoma of the breast]. PMID- 6296731 TI - [Hormonal receptors in malignant breast pathology]. PMID- 6296732 TI - [Neurological pathology in dialysis]. PMID- 6296733 TI - Double fluorescence labelling supports electrophysiological evidence for dichotomizing peripheral sensory nerve fibres in rats. AB - The technique of double labelling with fluorescent tracers has been used to demonstrate that some sensory neurones in the sixth lumbar dorsal root ganglion (L6) of the rat possess peripheral processes which dichotomize into both the pudendal and sciatic nerves. Bisbenzimide (Bb) was injected into the pudendal nerve and propidium iodide (PI) into the sciatic. Double labelled neurones were observed which fluoresced with a blue nucleus and orange cytoplasm (PI) at 405 nm excitation. The double fluorescence was confirmed by microspectrophotometric measurements. Injection of Nuclear Yellow into the sciatic nerve and Fast Blue into the pudendal nerve also produced double labelled neurones in L6. PMID- 6296734 TI - Nutrition classics. European Journal of Biochemistry, Volume 80, 1977: Mechanism of the prolyl hydroxylase reaction. 1. Role of co-substrates. PMID- 6296735 TI - Studies into postural hypotension in elderly patients. AB - Postural hypotension is common in elderly people. A study of 247 patients in Wellington showed an incidence of 33.1 percent. The causative factors included drugs such as diuretics and antihypertensive agents; the consequences of cerebrovascular accidents and peripheral neuropathies; and in a small number, the dysfunction of the autonomic nervous system. The test for autonomic function were difficult to apply to patients and any results likely to be unsatisfactory. PMID- 6296736 TI - Management of gestational trophoblastic disease: results of a cooperative study. AB - Three hundred fifty-four evaluable cases of hydatidiform mole diagnosed from 1970 to 1979 in 10 regional hospitals in Lombardy are analyzed in the present report. Twenty-six (7.3%) of the patients developed persistent trophoblastic disease. Younger (less than 20 years) and older (40 years or more) age at diagnosis, a large-for-dates uterus, and ovarian enlargement were associated with an increased risk of developing persistent trophoblastic disease. Twenty-three (9%) cases of persistent trophoblastic disease were observed among 250 women not prophylactically treated, but only in 3 (3%) among 104 who received prophylactic chemotherapy. High-risk groups are defined and the role of prophylactic chemotherapy is discussed. PMID- 6296737 TI - Glassy cell carcinoma of the cervix. AB - Of the 389 cases of primary adenocarcinoma of the cervix in the Armed Forces Institute of Pathology files, only 9 (2%) demonstrated features of glassy cell carcinoma in one third or more of the tumor. Histologically, glassy cell carcinomas were associated with admixed areas of mucinous adenocarcinoma in 5 instances, and 4 of the 9 neoplasms contained glandular differentiation by glassy cells. Squamous differentiation occurred in 4 of the 9 cases, and intracellular mucin was present in 7 of the 8 glassy cell carcinomas stained for it. Even in the single example without demonstrable mucin production gland formation by the tumor cells was evident. Although the cells have a characteristic appearance, it is unclear if glassy cell carcinoma is a meaningful entity. The need for its separate designation in the classification of cervical carcinoma is questionable. If glassy cell carcinoma exists, examples probably will be found within categories of nonkeratinizing large cell carcinoma and undifferentiated carcinoma. PMID- 6296738 TI - Gestational trophoblastic neoplasms in homozygous twins. PMID- 6296739 TI - Mullerian agenesis, hypoplasia of distal extremities, unusual dermatoglyphics, and pigmentation. AB - The case of a 17-year-old girl with mullerian agenesis, hypoplasia of the distal extremities, unusual dermatoglyphic findings, and mottled skin pigmentation is reported. The family history indicates that the maternal grandmother's sisters were similarly affected with mullerian agenesis, suggesting a dominant gene with variable penetrance in this family. PMID- 6296740 TI - Enzyme assays to detect preovulatory human luteinizing hormone surge. AB - Nonradioisotopic enzyme assays have been developed to detect the preovulatory luteinizing hormone (LH) surge in the unextracted urine of normally menstruating women. The amino groups on 6-mm glass beads were activated and coupled to the anti-LH-gamma-globulin for enzyme immunoassay or to the LH receptor prepared from bovine corpora lutea for enzyme receptorassay. The LH was conjugated with alkaline phosphatase. The enzyme assays detected preovulatory LH surge in the morning urine samples of 12 normally menstruating women. The LH surge was coincidental with preovulatory serum estradiol rise, shift in basal body temperature, and 4+ spinnbarkeit of the cervical mucus. The serum estradiol and progesterone levels in the luteal phase were indicative of corpus luteum formation. PMID- 6296741 TI - Cervical papillomavirus infection in diethylstilbestrol-exposed progeny. AB - Histologic sections from cervical and vaginal biopsies showing dysplasia in 37 women exposed to diethylstilbestrol (DES) in utero were stained for human papillomavirus structural antigens using an immunoperoxidase technique. Forty three percent of the lesions had detectable papillomavirus antigens. These findings indicate that a significant proportion of cervical dysplasia observed in DES-exposed progeny is associated with papillomavirus infection. PMID- 6296742 TI - Mechanisms of uterine bleeding in postmenopausal patients receiving estrogen alone or with a progestin. AB - The response of postmenopausal endometrium to cyclic estrogen and progestin and cyclic estrogen alone was studied in 75 biopsies and over 2000 preparations using standard histologic, histochemical, and scanning and transmission electron microscopic techniques. Estrogen and a progestin caused the atrophic endometrium to assume normal proliferative and secretory phases and to develop nucleolar channel systems. Cyclic unopposed estrogen produced unphysiologic responses in the glands, stromal cells, and vessels. The concept of a progestin or progesterone producing a "medical curettage" should be reappraised. Cyclic estrogen and progestin therapy do not cause all the endometrium to desquamate to the basalis layer. The combination therapy is associated with increased glycoprotein production in the gland and stromal cells, and an orderly regression and remodeling of the endometrium upon hormonal withdrawal. Cyclic estrogen alone causes irregular and unpredictable breakdown, which may or may not extend to the basalis. The stimulation of the endometrium by estrogen alone may allow the endometrium to use the majority of its energy for growth, which may lead to hyperplasia and neoplasia. PMID- 6296743 TI - Testosterone/androstenedione ratio in the evaluation of women with ovarian androgen excess. AB - A plasma testosterone/androstenedione (T/A) ratio was calculated in 15 women with possible ovarian neoplasms and androgen excess and whose mean plasma concentration of testosterone approached or was greater than a twofold elevation. The T/A ratio approached unity in eight of nine patients with polycystic ovarian disease, whereas all patients with hyperthecosis and two of three patients with androgen-secreting tumors had a testosterone/androstenedione ratio above 1.5. In 13 of 16 patients with androgen-secreting tumors of the ovary reported in the literature, the testosterone/androstenedione ratio was above 1.5. An elevated testosterone/androstenedione ratio should raise the suspicion of either an ovarian androgen-secreting tumor or hyperthecosis. PMID- 6296745 TI - Keyes technique questioned. PMID- 6296744 TI - Male pseudohermaphroditism due to 17 alpha-hydroxylase deficiency: diagnosis by gas chromatography--mass spectrometry. AB - A case of male pseudohermaphroditism associated with 17 alpha-hydroxylase deficiency is reported in which the diagnosis was firmly established by gas chromatography-mass spectrometry. The patient was a 20-year-old genotypic male, phenotypic female who presented with primary amenorrhea, absence of body hair, and no breast development. She was hypertensive. Corticosterone and progesterone levels were very high and sex steroids were virtually absent. Gonadotropins were in the menopausal range. The results of radioimmunoassay were not diagnostic. Analysis of urine using gas chromatography-mass spectrometry revealed a striking absence of steroids with 17-oxygen function-nor were there any 18 or 19 carbon steroids. These results clearly establish a total deficiency of 17 alpha hydroxylase activity. PMID- 6296746 TI - Enhancement of Lewis lung carcinoma by the concomitant infection of the host with herpes simplex virus type 1 and type 2. AB - Infection with herpes simplex virus (HSV) enhances the growth of Lewis lung tumor (LLT). Infection with HSV-1 enhances as efficiently as HSV-2. A significant acceleration of primary tumor formation was obtained. In mice, lung metastases were increased in number and size above the controls. Enhancement occurred only with high doses of infectious virus and was also obtained only when the virus infection was at a site close to that of the tumor implantation. This finding, in addition to the lack of antigenicity of LLT, is interpreted as arguing against an immunologic mechanism of the phenomenon. PMID- 6296747 TI - [Occurrence of photosensitivity in Wilms' tumor (malignant nephroblastoma)]. PMID- 6296749 TI - Eccrine porocarcinoma: a case report with light microscopy and ultrastructure. AB - A case of eccrine porocarcinoma in a 63-yr-old man is reported. There was a 10 yr history of an abdominal skin lesion which recurred after cautery and metastasized to skin and abdominal lymph nodes. Histologically, the primary tumour was composed of small squamoid cells with ductal lumina. There was intraepidermal nesting spread and an invasive dermal component. Formation of intracytoplasmic lumina in cells with squamoid features, best seen on electron microscopy, pointed towards poral differentiation and was a useful diagnostic feature which persisted in the less well differentiated metastases. The role of hidroacanthoma simplex as a possible preinvasive precursor lesion is discussed. The differential diagnosis from other skin tumours is described. The literature is reviewed. Despite their sometimes innocuous histological appearance, eccrine porocarcinomas are aggressive lesions which require wide local excision of the primary site and consideration of regional lymph node dissection. PMID- 6296748 TI - Necrotizing myopathy associated with steroid therapy. Report of two cases. AB - A range of histological lesions was found in necropsy samples of skeletal muscle from 2 patients on steroid therapy. There were perifascicular necroses, regeneration, atrophy, and sclerosis. These findings disagree with published data from human material; however, they are similar to observations made in experimentally induced steroid myopathy in rabbits. PMID- 6296750 TI - [Reduced glutathione and cytochrome C in the experimental therapy of shock]. PMID- 6296751 TI - Properties of vitamin E-deficient erythrocytes following peroxidant injury. AB - Several membrane properties of vitamin E-deficient and normal erythrocytes were studied after incubation of these cells with hydrogen peroxide. Measurements of mean corpuscular volume, cation permeability, membrane Na+, K+ ATPase activity, red cell filterability through 5 mu millipore filters, and membrane protein pattern on sodium dodecyle sulfate-gel electrophoresis revealed marked alterations before lysis. Vitamin E sufficient cells were unaffected by a similar incubation with hydrogen peroxide. We speculate that the changes in membrane function, which follow peroxidant injury, contribute to the shortened red cell survival in the vitamin E deficient state. PMID- 6296752 TI - Superoxide-dependent chemotactic activity for PMNs derived from opsonized zymosan stimulated human platelets. AB - Previous studies have shown that platelets exhibit a H2O2 producing, NADH dependent system that is activated by interaction with particulate material. Current evidence suggests that this system could be critically involved in th generation of chemotactic factor(s). In the present studies, chemotactic activity for polymorphonuclear leukocytes of supernatants derived from zymosan-stimulated human platelets has been evaluated using an agarose gel technique. Supernatants of opsonized zymosan-stimulated platelets showed significant chemotactic activity (migration index = 300 +/- 50), in comparison with supernatants prepared from platelet suspensions stimulated with nonopsonized-zymosan (migration index = 10 +/- 15) or resting platelet supernatants (migration index = 15 +/- 15). Furthermore, a marked increase in chemotactic activity of the opsonized zymosan treated platelet supernatants was demonstrated after the addition of NADH (migration index = 525 +/- 100). The inclusion of specific inhibitors of the cycloxygenase and lipoxygenase pathways resulted in a marked reduction of chemotactic activity, which was restored in the presence of NADH. Further, the addition of superoxide dismutase completely abolished the chemotactic response induced by NADH. These data suggest that platelets are the source of chemotactic factor(s) derived from the activation of a superoxide generating system. PMID- 6296753 TI - Effect of amiloride on potential difference across rectal mucosa in cystic fibrosis patients. AB - This study investigated whether the altered epithelial cell function in cystic fibrosis (CF) concerns channels for passive diffusion of sodium across cell membranes. For this purpose, the potential difference (PD), which arises across the colonic mucosa from electrogenic sodium transport, was compared in CF and non CF subjects, together with the effect on this PD from bathing the mucosa with the specific Na+ channel blocker, amiloride. The resting rectal mucosal PD did not differ significantly in the two groups. Amiloride caused a dose-dependent decrease in the PD (lumen negative to skin) in all subjects. The mean amiloride effect was less in the CF population at 10(-8), 10(-7), 10(-6), 10(-5), and 10(4) M amiloride. The decrease of transmucosal PD in CF patients was significantly less when compared with controls at 10(6) M and 10(-5) M and suggestive of a similar trend at 10(-4) M. PMID- 6296754 TI - Recurrent polyneuropathy with multiple herpesvirus infections. AB - A 4-year-old boy developed typical acute polyneuritis at 2 years of age and returned to fully normal status in 2 months. When he was 4 years old, an identical neurologic syndrome recurred at the same time that his sister was determined to have classic infectious mononucleosis. Both siblings then had positive Monospot tests. The boy with recurrent symmetrical neuropathy had an initial Epstein-Barr virus-viral capsid antibody (EBV-VCA) serum titer of 1:160 and this dropped to 1:40 after 1 month. A persistent complement fixation titer to cytomegalovirus (CMV) of 1:64 was measured in his serum and his concentrated CSF contained a CMV complement fixation antibody titer of 1:32, a rare finding in an age-matched control group. Cultures of his urine, buffy coat, and saliva for CMV were negative. Electrophoresis of his CSF revealed oligoclonal bands, several of which were also present in his serum. It seems most likely, but not proven, that the clinical paralysis was caused initially by a CMV infection and that the Epstein-Barr virus infection reactivated the latent CMV to produce the recurrent weakness. PMID- 6296755 TI - Evaluation of the pertussis components of diphtheria-tetanus-pertussis vaccine. AB - The adjuvant and antigen components of the pertussis fraction of diphtheria tetanus-pertussis (DTP) vaccine were evaluated. Four preparations of DTP vaccine composed of either whole cell (Wc) or extracted (E) pertussis antigen combined with either an aluminum phosphate (Ph) or alum (Al) adjuvant were compared. Local reactions were similar in all four vaccines after the first two immunizations but were significantly increased in incidence and severity following the third immunization with vaccine WcPh. This appeared to be due to the Ph adjuvant rather than the antigen component. Febrile reactions were experienced more often (P = .0009) and with higher temperatures (P = .0001) with the WcPh vaccine following the first immunization. This appeared to be due to the Wc component. Comparing the pooled Wc groups with the pooled E groups revealed a greater febrile response in the Wc group after both the first (P = .0008) and the second (P = .03) immunization. Local reactions appear temporally and etiologically to be distinct from febrile reactions. The pooled Wc antigen group produced a higher geometric mean titer than the pooled E antigen group (P = .05). Serologic responses, with respect to geometric mean titer, were not significantly different among the individual vaccines. This study suggests that the combination of whole cell and aluminum phosphate, which is currently in use in the United States, is probably not the optimal formulation for pertussis vaccine. PMID- 6296756 TI - [Acropapulovesicular syndrome in children and Gianotti-Crosti disease. Discussion apropos of 4 cases of probable viral etiology]. PMID- 6296757 TI - Leukotrienes, allergy and inflammation. AB - Immunological and non-immunological injury induce as a result of the action of the enzyme lipoxygenase the release of a series of arachidonic acid metabolites known as leukotrienes. The leukotrienes play an important role in allergic and inflammatory disease. Leukotrienes C4, D4 and E4 which recently have been recognized as constituents of the allergic mediator slow reacting substance of anaphylaxis (SRS-A) induce powerful bronchoconstriction, plasma exudation and weal and flare responses. Leukotriene B4 is involved in the regulation of chemotaxis, chemokinesis and other aspects of both cellular and vascular inflammation. The development of specific lipoxygenase inhibitors may lead to a new class of drugs for the treatment of bronchial asthma and chronic inflammatory diseases. PMID- 6296758 TI - Sodium action potentials induced by calcium chelation in rat uterine smooth muscle. PMID- 6296759 TI - The location and properties of preganglionic vagal cardiomotor neurones in the rabbit. AB - The origins of preganglionic vagal neurons which slow the heart in the rabbit have been examined with standard neurophysiological stimulation and recording techniques. The activity of 216 neurones projecting to the right cervical vagus nerve have been recorded in localized areas of the brain stem. Thirty-six of these neurones were classified as cardiomotor neurones since they had properties similar to those described for such neurones in the cat. All had efferent axons in the range of B fibres. They could be synaptically activated by electrical stimulation of the ipsilateral aortic nerve which in the rabbit is solely barosensory. The majority of these neurones (70%) were spontaneously active and those which were normally silent could be made to fire by iontophoretic application of DL-homocysteic acid (an excitant amino acid). This spontaneous, or evoked, activity showed evidence of a pulse rhythm (of baroreceptor origin) and respiratory modulation (firing predominantly during expiration). In response to application of DL-homocysteic acid, the neuronal excitation was usually accompanied by a small but significant bradycardia. Histological examination showed that these neurones were located in both the dorsal vagal nucleus and the nucleus ambiguus. PMID- 6296760 TI - The automated analysis of data from single ionic channels. AB - The development of single channel recording has brought with it the need to analyse enormous amounts of data. The data analysis is time consuming and subject to observer biases since the events are random in time and are contaminated with uncorrelated noise. We have developed a heuristic pattern recognition program which identifies with high precision single channel currents and rejects contaminating noise. The program interactively provides for a variety of amplitude and duration measures. Analysis is flexible and rapid: a file containing over 10,000 events can be analysed in under 2 h. Specific detection features include variable lowpass filtering, automatic baseline restoration, and adaptive amplitude thresholds. A record is analysed through duration histograms, binomial estimates of the number of active channels present, cross-correlation estimates between parameters, spectral analysis of events and background noise, and stationarity of mean channel current. The graphic output facilities can plot raw data (after filtering and baseline restoration) with the idealized signal superimposed or with detected events underlined. A batch processing facility has been included to allow processing of data during periods of low computer demand. PMID- 6296761 TI - [Kaposi's disease. 2 recent cases]. AB - Known since 1872, the multifocal angiosarcoma called Kaposi's sarcoma takes different forms in different communities. It develops slowly among the mediterranean populations and Jewish Central Europe and most rapidly among Bantus, in Africa, and white homosexuals. Another aspect of the disease has been described in patients with transplanted kidney, where it seems to be associated with immunosuppressive treatments. The recent outbreak of Kaposi's sarcoma among homosexuals in New York and San Francisco raises new and difficult problems concerning its aetiology, as the outbreak cannot be entirely explained by pressure from sexually transmissible diseases. Having observed two new cases in France, the authors have attempted to re-evaluate the pathophysiological factors of the disease, the most constant of which seems to be a considerable degree of immunodepression, perhaps partly due to chronic infection. PMID- 6296762 TI - [Assay of cefotaxime in the middle ear secretions of serous otitis in children]. PMID- 6296763 TI - [Peroperative rupture of the lumbar dura mater with nerve root hernia. Management]. AB - Rupture of the dura mater during lumbar laminectomy may result in destruction of the herniated sacral and lumbar nerve roots. This can be avoided by a simple but immediate surgical procedure, details of which are presented here. PMID- 6296764 TI - A general strategy for cloning double-stranded RNA: nucleotide sequence of the Simian-11 rotavirus gene 8. AB - Using Simian-11 rotavirus RNA, a strategy has been developed for the production of full length cloned copies of the genes of double-stranded (dsRNA) viruses. Genomic RNA segments were polyadenylated and reverse transcribed to yield a mixture of full length cDNA copies of both possible polarities. The cDNAs were annealed, filled in to complete any partial copies, tailed and inserted into the PstI site of pBR322 using dG/dC tailing. Cloned rotavirus cDNA gene copies were assigned to genomic RNA segments by Northern hybridization. The complete sequence of gene 8 which codes for NCVP3, a non-structural protein of SA11 rotavirus, was determined from a cloned gene copy. It is 1059 bases in length and has an open reading frame which could code for a protein containing 317 amino acids. The apparent 5' and 3' terminal non coding regions are 46 and 59 bases in length, respectively. The sequence ATGTGACCOH at the 3' end of the plus strand is conserved in four of the eleven genes examined. The cloning procedures used should be generally applicable to viruses with segmented dsRNA genomes. PMID- 6296765 TI - In vitro accurate initiation of transcription on the adenovirus type 2 IVa2 gene which does not contain a TATA box. AB - In vitro initiation of transcription has been studied on the IVa2 gene of the adenovirus type 2 ( Ad2 ) which is expressed at intermediate time after infection. This gene does not contain a TATA box around 30 bp upstream from the cap site. By analyses of the in vitro runoff products by size and by single strand nuclease protection mapping, it was concluded that the in vitro initiation occurred accurately at two in vivo cap sites with low but detectable efficiency. By using cloned IVa2 DNA, which is free from the major late promoter, it was suggested that the low efficiency of the in vitro initiation on the IVa2 gene may not be due to close location of the two genes but may rather reflect the efficiency of the promoter itself. The cell-free extracts from both uninfected and infected HeLa cells supported the accurate initiation of transcription in vitro with almost the same efficiency. In addition, the cell free extract directed an artificial initiation of transcription in vitro by recognizing a TATA box located 140 bp upstream from the cap sites. PMID- 6296766 TI - Sequence dependent interaction of hnRNP proteins with late adenoviral transcripts. AB - Irradiation with ultraviolet light was used to induce covalent linkage between hnRNA and its associated proteins in intact HeLa cells, late after infection with adenovirus type 2. Covalently linked hnRNA-protein complexes, containing polyadenylated adenoviral RNA, were isolated and their protein moiety characterized. Host 42,000 Mr hnRNP proteins proved to be the major proteins crosslinked to viral hnRNA. To investigate their possible involvement in RNA processing, the localization of these cross-linked polypeptides on adenoviral late transcripts was determined. Sequences of RNA around the attachment sites of the protein were isolated. After in vitro labeling they were hybridized to Southern blots of adeno DNA fragments. The hybridization patterns revealed that the 42,000 Mr polypeptides can be linked to adenoviral transcripts over the entire length of the RNA, corresponding to 16.2-91.5 m.u. of the viral genome. Fine mapping within the Hind III B region (16.8-31.5 m.u.) established, however, that the localization of the cross-linked polypeptides was not random in all parts of the transcript. Sequences around the third leader and the 3' part of the i-leader were overrepresented, whereas the regions encoding VA I and VA II RNA and the late region 1 mRNA bodies were underrepresented in the cross-linked RNA. Using genomic DNA fragments and a cDNA clone containing the tripartite leader it appeared that leader and intervening sequences were represented about equally in cross-linked RNA fragments. Although these results do not support the notion that introns or exons are specifically interacting with one RNP protein, they demonstrate that the 42,000 hnRNP proteins are non randomly positioned on the RNA sequence. PMID- 6296767 TI - Cloning and nucleotide sequencing of the bovine growth hormone gene. AB - A gene coding for bovine growth hormone was isolated from a bovine genomic library. The nucleotide sequence of the coding regions of the gene was found to be identical with that of a nearly full-length growth hormone cDNA clone. The gene sequence is approximately 1800 bp in length and contains four intervening sequences. The second intervening sequence of 227 nucleotides does not contain a repetitive element similar to that observed in the rat growth hormone gene. A comparison of the 5' and 3' flanking and untranslated regions of the bovine, human and rat growth hormone genes revealed many areas of highly conserved sequence. Especially noteworthy was the observation that all three genes had a 38 nucleotide homologous sequence within their 5' flanking regions located about 100 bp upstream from their transcription initiation sites. PMID- 6296768 TI - Studies on the biological role of DNA methylation: V. The pattern of E.coli DNA methylation. AB - The distribution of the methylatable sites GATC and CCATGG was studied by analyzing the molecular average size of restriction fragments of E. coli DNA. Both sites were found to be randomly distributed, reflecting a random pattern of methylation. The methylation pattern of specific sequences such as the origin of replication and rRNA genes has been studied in wild type E. coli and a methylation deficient (dam- dcm-) mutant. These sequences were found to be methylated in wild type cells and unmethylated in the mutant indicating that there is no effect of the state of methylation of these sequences on their expression. Analysis of the state of methylation of GATC sites in newly replicating DNA using the restriction enzyme Dpn I (cleaves only when both strands are methylated) revealed no detectable hemimethylated DNA suggesting that methylation occurs at the replication fork. Taking together the results presented here and previously published data (5), we arrive at the conclusion that the most likely function of E. coli DNA methylations is probably in preventing nuclease activity. PMID- 6296769 TI - Deletions at intervening sequence splice sites in the alcohol dehydrogenase gene of Drosophila. AB - Two formaldehyde-induced, homozygous viable ADH-negative mutants, Adhfn4 and Adhfn6, possess no material that cross-reacts with antibody directed against ADH, no mature mRNA of wild-type size, and greatly reduced amounts of RNA that hybridizes with an Adh probe. We have cloned the genomic DNA sequences from these mutants in bacteriophage lambda Charon 4 and subcloned the Adh region into plasmid vector pBR327. Restriction analyses revealed one small deletion in each of these mutants and DNA sequencing showed that the splice junctions of the 65 base pair (bp) intervening sequence (IVS) were altered. Both cloned mutant Adh genes, as well as the wild-type gene, are capable of promoting correct specific transcription initiation in HeLa cell nuclear extracts in vitro. We conclude that Adhfn4 and Adhfn6 are defective in RNA processing. Our results provide evidence for the importance of the splice junction sequences in normal ADH RNA processing and stabilization in Drosophila. We also speculate that splicing of ADH RNA proceeds in a nonrandom manner: mutations in one of the intervening sequences appear to cause accumulation of a large ADH RNA containing at least one other IVS. PMID- 6296770 TI - Selective binding of actinomycin D and distamycin A to DNA. AB - The exact sites at which a number of drugs inhibit the nick translation of DNA by E.coli DNA polymerase-I have been pinpointed. In order to do this, a method has been developed for sequencing double-stranded plasmid DNA from the site of a specifically induced nick. The initial experiments have concentrated on analysis of drug inhibition of nick translation in a 200 nucleotide region near the Eco Rl origin of pBR313. Many drugs were found to inhibit nick translation in a highly sequence specific manner. For actinomycin D, significant inhibition occurred at just four sites in the nucleotide sequence under test and only one sequence (pGpCpGpCpGpGp) gave really strong inhibition. Distamycin A gave a different pattern of inhibition with particularly strong stops in just two of the many A-T rich regions in the DNA. Experiments with caffeine suggest that factors in addition to primary sequence are important in determining where major inhibition occurs. PMID- 6296771 TI - Mapping of RNA polymerase binding sites in R12 derived plasmids carrying the replication-incompatibility region and the insertion element IS1. AB - Interactions between Escherichia coli RNA polymerase holoenzyme and three small plasmid DNAs (pSM1, pSM2, and pSM15) derived from the drug resistant factor R12 have been studied. These plasmids carry the copy number and incompatibility determinants, the origin of DNA replication and the rep gene(s) necessary for plasmid replication. They also contain the insertion element IS1 and the putative finO cistron. Thirteen DNA segments within the largest of the three plasmids (pSM2) were able to form either a binary and/or ternary complex with RNA polymerase. A unique strong binding site was mapped within the left end of IS1. Five binding sites were found within the rep-cop-inc region. Four of these are weak binding sites whereas the fifth does not form a stable binary complex and was detected by ternary complex formation. A strong binding site was located in the putative finO region whereas the remaining six binding sites are located in regions with unidentified genetic functions. PMID- 6296772 TI - Nucleotide sequence of a segment of human DNA containing the three tRNA genes. AB - A 1.65 kb segment of DNA from a human-lambda recombinant, selected for human tRNA genes, was subcloned in the plasmid pAT 153 for sequence determination. Three human tRNA genes were found; one, a tRNAlys gene which appears to specify a tRNA identical in sequence to the previously described tRNA3lys from rat liver (AAA,G); another a tRNAgln gene, the product of which would be expected to recognize only the codon CAG; and a third which would direct synthesis of a tRNAleu specific for CUA and G. Intervening sequences are not found in any of these genes. The three tRNA genes are separated by about 0.5 kb segments of DNA containing no apparent informational sequences. Examination of flanking sequences shows some similarities at the 5'-ends of the three genes, but less similarity to 5' flanking sequences of tRNA genes in other eucaryotes. All three tRNA genes direct synthesis of appropriate sized products in a HeLa cell lysate in vitro transcription system. PMID- 6296773 TI - Transcription initiation site of rat ribosomal DNA. AB - The sequence of 1,100 nucleotides surrounding the transcription initiation site of a cloned rat ribosomal RNA gene (rDNA) has been determined. The location of the 5' terminus of 45S pre-rRNA was determined by S1 nuclease mapping, reverse transcriptase elongation and confirmed by in vitro capping of 45S rRNA and in vitro transcription. Two different plasmid subclones, from two separate genomic clones of rat rDNA, contained the identical sequence surrounding the transcription initiation site: -10GGAGATATAT 1GCTGACACGC TGTCCTTTTG+20. Relatively long, greater than 15 base pairs, regions of sequence homology were found when the sequences of the initiation regions of rat and mouse rDNA (Urano, Y., Kominami, R., Mishima, Y., and Muramatsu, M., Nucleic Acids Res. 8, 6043 6058, 1980) were compared. When both the rat and mouse sequences were compared to that of human rDNA (G. Wilson, personal communication) a sequence of 15 nucleotides immediately following the initiation sites were found to be preserved. PMID- 6296774 TI - The nucleotide sequence of the dnaA gene and the first part of the dnaN gene of Escherichia coli K-12. AB - The nucleotide sequence of the dnaA gene and the first 10% of the dnaN gene was determined. From the nucleotide sequence the amino acid sequence of the dnaA gene product was derived. It is a basic protein of 467 amino acid residues with a molecular weight of 52.5 kD. The expression of the dnaA gene is in the counterclockwise direction like the one of the dnaN gene, for which potential startsites were found. PMID- 6296775 TI - Evidence that rifampicin can stimulate readthrough of transcriptional terminators in Escherichia coli, including the attenuator of the rpoBC operon. AB - The genes encoding the beta and beta' subunits of RNA polymerase in E.coli, rpoB and rpoC, lie downstream of at least two ribosomal protein genes, rplJ (encoding L10) and rplL (L7/12), in a common operon. All four genes are served by promoter PL10, and an attenuator (partial terminator) of transcription, t1, lies between rplJL and rpoBC. Treatment of E.coli with rifampicin, under conditions producing partial inhibition of general RNA synthesis, can stimulate transcription of rpoBC. We have investigated the locus of this effect by fusing PL10 and t1 separately to galK, in suitable plasmids. Our studies of these fusions, and similar fusions involving transcriptional terminators derived from coliphage T7, indicate that low concentrations of rifampicin cause increased readthrough of several different transcriptional terminators in E.coli in vivo, including rpo t1. We discuss whether or not this unspecific mechanism is solely responsible for the observed stimulatory effects of the drug on rpoBC transcription. PMID- 6296776 TI - Effect of rifampicin on expression of lacZ fused to promoters or terminators of the E.coli rpoBC operon. AB - The genes encoding the beta and beta' subunits of RNA polymerase in E.coli lie downstream of at least two ribosomal protein genes in a single unit of transcription. Treatment of E.coli with rifampicin, under conditions producing partial inhibition of general RNA synthesis, can strongly stimulate transcription of the polymerase genes, while activating the neighbouring ribosomal genes only slightly. We have investigated the mechanism of this effect by fusing strong promoters, a weak internal promoter, and an attenuator of the polymerase operon to the lacZ gene, in derivatives of plasmid pMC81. Studies of these fusions confirm our conclusion, based on similar fusions to galK, that rifampicin can foster readthrough of transcriptional terminators. They also suggest the existence of extra terminators and anti-termination elements in the above transcription unit. PMID- 6296777 TI - Silica gel: an improved support for the solid-phase phosphotriester synthesis of oligonucleotides. AB - The phosphotriester method for the stepwise synthesis of deoxyoligonucleotides has been employed using HPLC-grade silica gel (Porasil B) as the solid support. The procedure results in a convenient flow-through system for the synthesis of oligomers where all the reaction steps including the zinc bromide method of detritylation are compatible with the selected support. Deoxyoligonucleotides of 25-30 nucleotides in length can be synthesized in high yields utilising stable phosphotriester intermediates. Ease of handling of the solid support allows convenient synthesis of mixed oligonucleotide sequences. PMID- 6296778 TI - Sequence of the malK gene in E.coli K12. AB - We present the sequence of gene malK which encodes a component of the system for maltose transport in E.coli K12. We also determined the position of deletion (S50) which fuses malK to the following gene lamB; the malK-lamB protein hybrid contains all of the malK protein. The mRNA corresponding to the last two thirds of gene malK could form stable stem and loop structures. The malK protein, as deduced from the gene sequence, would include 370 residues and correspond to a molecular weight of 40700. The sequence as well as sequence comparisons with the ndh protein of E.coli are discussed in terms of the location and function of the malK protein. PMID- 6296779 TI - Ubiquitous transposon-like repeats B1 and B2 of the mouse genome: B2 sequencing. AB - Mouse genome contains two major families of short interspersed repeats in more than 10(5) copies scattered throughout the whole genome. They are referred to as B1 and B2 sequences since they were first isolated from the genome library by means of a dsRNA-B probe /1/. In this work, two copies of the B2 family were sequenced and compared with the previously sequenced B1 repeat /2/. A B2 ubiquitous repeat is ca. 190 bp long. The members of the family deviate in 3-5% of nucleotides from the consensus sequence. B2 contains regions of homology to the RNA polymerase III split promoter and to 4.5S snRNA I. Both B1 and B2 contain regions which resemble junctions between exons and introns. In contrast to B1, B2 does not contain apparent homologies to papova viral replication origins and a human Alu sequence. One side of the B2 repeat is represented by a very AT-rich sequence (ca. 30 bp long) followed with an oligo (dA) stretch 10-15 nucleotides long. This region of the repeat is the most variable one. The whole unit is flanked with 15-16 bp direct repeats different in sequenced copies of B2. The same is true of some copies of the B1 family. The properties of B1 and B2 repeats suggest that they may represent a novel class of transposon-like elements in eukaryotic genome. A possible role of B-type repeats in genome reorganization, DNA replication and pre-mRNA processing is discussed. PMID- 6296780 TI - Nucleotide sequence of satellite DNA contained in the eliminated genome of Ascaris lumbricoides. AB - Several restriction endonuclease fragments isolated from highly repetitive satellite DNA of the chromatin eliminating nematode Ascaris lumbricoides var. suum have been cloned. Each type of restriction fragment corresponds to a different variant of the same related ancestral sequence. These variants differ by small deletions, insertions and single base substitutions. Restriction and DBM blot analyses show that members of the same variant class are tandemly linked and therefore are physically separated from other variant classes. A comparison of all the determined sequences establishes a 121 bp long and AT rich consensus sequence. There is evidence for an internal short range periodicity of 11 bp length, indicating that the Ascaris satellite initially may have evolved from an ancestral undecamer sequence. The satellite DNA sequences are mostly but not entirely eliminated from the presumptive somatic cells during chromatin diminution. We have no evidence for transcriptional activity of satellite DNA at any stage or tissue analyzed. PMID- 6296781 TI - Nucleotide sequences of tobacco chloroplast genes for elongator tRNAMet and tRNAVal (UAC): the tRNAVal (UAC) gene contains a long intron. AB - The nucleotide sequences of tobacco chloroplast genes for elongator tRNAMet and tRNAVal (UAC) have been determined. The tRNAVal gene contains a 571 base pairs intron located in the anticodon loop. The tRNAVal gene is transcribed as a 750 bases precursor RNA molecule. Both tRNAs deduced from the DNA sequences show 97% sequence homologies with those of spinach chloroplasts. PMID- 6296782 TI - Characterization of histone genes isolated from Xenopus laevis and Xenopus tropicalis genomic libraries. AB - Using a cDNA clone for the histone H3 we have isolated, from two genomic libraries of Xenopus laevis and Xenopus tropicalis, clones containing four different histone gene clusters. The structural organization of X. laevis histone genes has been determined by restriction mapping, Southern blot hybridization and translation of the mRNAs which hybridize to the various restriction fragments. The arrangement of the histone genes in X. tropicalis has been determined by Southern analysis using X. laevis genomic fragments, containing individual genes, as probes. Histone genes are clustered in the genome of X. laevis and X. tropicalis and, compared to invertebrates, show a higher organization heterogeneity as demonstrated by structural analysis of the four genomic clones. In fact, the order of the genes within individual clusters is not conserved. PMID- 6296784 TI - Sequence of the chloroplast 16S rRNA gene and its surrounding regions of Chlamydomonas reinhardii. AB - The sequence of a 2 kb DNA fragment containing the chloroplast 16S ribosomal RNA gene from Chlamydomonas reinhardii and its flanking regions has been determined. The algal 16S rRNA sequence (1475 nucleotides) and secondary structure are highly related to those found in bacteria and in the chloroplasts of higher plants. In contrast, the flanking regions are very different. In C. reinhardii the 16S rRNA gene is surrounded by AT rich segments of about 180 bases, which are followed by a long stretch of complementary bases separated from each other by 1833 nucleotides. It is likely that these structures play an important role in the folding and processing of the precursor of 16S rRNA. The primary and secondary structures of the binding sites of two ribosomal proteins in the 16SrRNAs of E. coli and C. reinhardii are considerably related. PMID- 6296783 TI - Organization and expression of cloned histone gene clusters from Xenopus laevis and X. borealis. AB - We have isolated several clones containing Xenopus histone genes from genomic libraries of X. laevis and X. borealis DNA. Each genomic clone has been mapped and the positions of 26 histone genes in seven laevis clones and 5 histone genes in one borealis clone have been determined. In laevis, the histone gene clusters show considerable variation in gene order within a single individual. When the cloned DNAs were microinjected into the nucleus of Xenopus oocytes, expression of cloned genes at the transcriptional and translational level was readily detectable. Previously unknown histone variants were revealed by the microinjection experiments. PMID- 6296785 TI - The relation between polyoma T-antigen and increased 5S RNA synthesis in cell free extracts from polyoma-infected mouse kidney cell cultures. AB - In polyoma-infected mouse kidney cell cultures 5S RNA synthesis began to increase around 16 h, i.e. 7-9 h after the onset of polyoma T-antigen synthesis. The rate of polyoma-induced 5S RNA synthesis reached a maximum plateau around 25 h when it was 1.8-2.0 times higher than in mock-infected parallel cultures. Stimulation of 5S RNA synthesis in vivo thus coincided in time with the increase in total cellular RNA and protein. Cell-free extracts (S100) prepared at 15 h from mock (S100-M) or polyoma-infected (S100-Py) mouse kidney cell cultures were indistinguishable with respect to protein concentration and 5S RNA synthesis, using a cloned somatic Xenopus borealis 5S gene as template. S100-Py extracted 25 h after infection contained 30% more protein and synthesized 1.5-2.0 times more 5S RNA than S100-M. Complete removal of the polyoma T-antigens from S100-Py by 3 cycles of immunoprecipitation with hamster anti-T serum remained without effect on stimulated 5S RNA synthesis. However, a linear relationship between 5S RNA synthesis and protein concentration of S100-M and S100-Py was observed. PMID- 6296786 TI - Spacer size heterogeneity in ribosomal DNA of Chironomus thummi is due to a 120 bp repeat homologous to a predominantly centromeric repeated sequence. AB - The rDNA of Ch. tepperi is homogeneous in structure with a repeat size of 8.4 kb. This size seems to be typical for the basic repeat unit in Chironomus species. Ch. th. piger rDNA cistrons are slightly increased in length (9.0 kb). In the non transcribed spacer (NTS) an appr. 0.18 kb segment is additionally present in about 50% of the repeats. Ch. th. thumni DNA contains largely heterogeneous rDNA repeats, mainly between 10 and 16 kb. The heterogeneity is due to varying numbers of 120 bp elements present in the NTS. The different spacer size classes are not randomly distributed. The short repetitive 120 bp elements (Cla I elements) hybridize in situ with the nucleolus and with centromere regions. The Cla I elements are regularly present in the thummi NTS, but are absent in the piger NTS. Only very few piger rDNA cistrons may contain Cla I elements. PMID- 6296788 TI - A short interspersed repetitive element found near some mouse structural genes. AB - We have isolated and characterized a family of interspersed repetitive elements which make up about 1% of the mouse genome. The elements represent a group of homologous but non-identical units about 400 bp in length. Individual members of the family show considerable divergence from one another. The spacial relationships between members of the family and a number of other identified mouse sequences including structural genes have been determined; these elements are found on the 5' as well as 3' sides of various genes at distances ranging from less than 1 to 7.5 kilobases (Kb). The sequences are present in the DNA of all species of Mus. Related sequences are present in the rat genome at a repetition frequency similar to that in the mouse genome. A partial sequence of one member of the family is presented. PMID- 6296787 TI - Escherichia coli delta 1-pyrroline-5-carboxylate reductase: gene sequence, protein overproduction and purification. AB - The sequence of the Escherichia coli proC gene which encodes for delta 1 pyrroline-5-carboxylate (PCA) reductase was determined. Overproduction of the proC gene product via an expression plasmid carrying the bacteriophage lambda PL promoter allowed the purification to homogeneity of PCA reductase by affinity adsorption chromatography. NH2 and COOH-terminal analysis and amino acid composition of the purified proC protein is consistent with the gene sequence reported. The molecular weight of the proC monomer is 28,112. PMID- 6296789 TI - An immunoglobulin promoter region is unaltered by DNA rearrangement and somatic mutation during B-cell development. AB - The V1 gene encodes the heavy chain variable region of antibodies that bind to phosphorylcholine in the Balb/c mouse. V1 genes have been cloned from mouse sperm DNA, an IgM-producing tumor HPCM2 and an IgA-producing tumor M167. The transcription start site of the V1 gene has been mapped 63 +/- 1 base pairs from the coding sequence for both alpha and mu transcripts. Comparison of flanking DNA sequence 574 base pairs 5' to the V1 transcription start site in sperm, HPCM2 and M167 DNA reveals that sperm and HPCM2 sequences are completely identical in this region and the M167 sequence differs from them by a single base change. Although the coding region of the V1 gene has undergone a high (4%) rate of somatic mutation in M167 we demonstrate that the somatic mutation mechanism stops near the transcription start site. These results demonstrate that initiation of V1 gene transcription remains unchanged with respect to location and 5' sequences throughout B-cell development. PMID- 6296790 TI - A switch region inversion contributes to the aberrant rearrangement of a mu immunoglobulin heavy chain gene in MPC-11 cells. AB - We describe the unique features of an aberrantly rearranged mu immunoglobulin heavy chain gene isolated from MPC-11 cells (a gamma 2b producing Balb/c plasmacytoma). A novel rearrangement has occurred 1.5 Kb 5' of the MPC-11 mu gene (denoted 18b mu) resulting in the deletion of the majority of the repetitive switch region (S mu) and 5' flanking DNA including the Joining (JH) sequences. The remainder (275 bp) of the S mu repeat has undergone a complete sequence inversion. DNA sequences 5' of the inverted S mu sequence do not resemble Variable (VH), Diversity (D), JH or their conserved flanking sequences. A DNA sequence localized 5' of the inverted S mu sequence, (p18b mu-1.4) detects a small family of homologous sequences in Balb/c DNA. The 18b mu-1.4 like sequences lack homology to S mu, exhibit flanking sequence polymorphisms in 5 out of 6 inbred mouse strains and undergo partial or complete deletion in 5 out of 10 plasmacytomas tested. Two 18b mu-1.4 homologous sequences display a higher copy number in C57Bl/6, AL/N and CAL9 mouse strains. PMID- 6296791 TI - The primary structure of the Saccharomyces cerevisiae gene for 3-phosphoglycerate kinase. AB - The DNA sequence of the gene for the yeast glycolytic enzyme, 3-phosphoglycerate kinase (PGK), has been obtained by sequencing part of a 3.1 kbp HindIII fragment obtained from the yeast genome. The structural gene sequence corresponds to a reading frame of 1251 bp coding for 416 amino acids with no intervening DNA sequences. The amino acid sequence is approximately 65 percent homologous with human and horse PGK protein sequences and is in general agreement with the published protein sequence for yeast PGK. As for other highly expressed structural genes in yeast, the coding sequence is highly codon biased with 95 percent of the amino acids coded for by a select 25 codons (out of 61 possible). Besides structural DNA sequence, 291 bp of 5'-flanking sequence and 286 bp of 3' flanking sequence were determined. Transcription starts 36 nucleotides upstream from the translational start and stops 86-93 nucleotides downstream from the translational stop. These results suggest a non-polyadenylated mRNA length of 1373 to 1380 nucleotides, which is consistent with the observed length of 1500 nucleotides for polyadenylated PGK mRNA. A sequence TATATATAAA is found at 145 nucleotides upstream from the translational start. This sequence resembles the TATAAA box that is possibly associated with RNA polymerase II binding. PMID- 6296792 TI - The 3' untranslated regions of two related mRNAs contain an element highly repeated in the sea urchin genome. AB - Two closely related cDNA clones, pSpec1 and pSpec2, specifying two developmentally regulated tissue specific mRNAs from sea urchin embryos were used to probe a sea urchin genomic lambda library. Screening 10,000 phage by plaque hybridization yielded several hundred positive signals. With more stringent wash procedures, only two to three phage were positive. Three of these phage, one isolated by stringent wash procedures and two isolated by standard wash procedures were further investigated by restriction analysis, RNA gel blots, and DNA sequencing. The phage isolated by the stringent wash procedure appears to be a gene coding for the Specl mRNA. The other phage contain only partial homology to pSpec1 and pSpec2, 150 to 200 base pairs of the 3' untranslated region of the Spec1 and Spec2 mRNAs. It is concluded that the Spec1 and Spec2 mRNAs contain a highly repetitive element near their 3' end. The element is present at 2000 to 3000 copies per genome and may be transcribed at some sites other than those coding for the Spec1 and Spec2 genes. The possible function and evolutionary origin of the repetitive element is discussed. PMID- 6296793 TI - Structure of a human beta-actin-related pseudogene which lacks intervening sequences. AB - From a human genomic library we have isolated and sequenced a beta-actin-related pseudogene (Hbeta Ac-psi l) which is free of intervening sequences. Several nucleotide insertions and deletions and translational stop codons generated within the protein-coding region indicate that this gene is functionless. PMID- 6296794 TI - Independently evolving chicken histone H2B genes: identification of a ubiquitous H2B-specific 5' element. AB - The DNA sequence of two chicken histone H2B genes has been determined. Both genes code for the same H2B subtype. Except for conserved "promoter" elements, the sequences 5' to the protein coding regions are completely divergent, indicating that the genes are distantly related and are not evolving in concert. This presents an ideal situation for sequence comparisons. We have discovered a 13 bp, H2B specific homology block, 5' CTCATTTGCATAC 3' located close to the "TATA box". This motif is conserved in all H2B gene leader regions so far sequenced. One of the H2B genes is closely linked, in a divergent arrangement, to an H2A gene, and sequence data suggests that the linked genes share promoter elements. PMID- 6296795 TI - Methods and validity of dietary assessments in four Scandinavian populations. AB - Average intakes of nonstarch polysaccharides (dietary fiber), foods, and nutrients were measured in representative samples of 30 men aged 50-59 in 4 Scandinavian populations with a 3-4 fold difference in risk for large bowel cancer. The assessment technique, a 4-day weighed record of food consumed and duplicate collections of all food eaten, was validated by chemical analysis of the duplicates, by measuring 24-hour urine and fecal nitrogen excretion, and by comparing the constituents of the urine samples collected during the survey with similar collections 1-2 weeks later. There were good agreements between estimates of fat and protein intake obtained by food-table calculations of the 4-day weighed record and the chemically analyzed duplicates. Urinary plus fecal nitrogen excretion was equal to estimated nitrogen intake during the survey, and no discernable changes in urinary output occurred after the survey, thereby implying that dietary habits had not changed as a result of the investigative technique. It is concluded that the dietary data are indicative of current patterns of food consumption and are sufficiently valid for comparison with data on cancer risk in the 4 areas. PMID- 6296796 TI - Dietary patterns in Them and Copenhagen, Denmark. AB - Four-day weighing and 24-hour recall were used to record food consumption in groups of 30 men, aged 50-59 years, in 2 areas of Denmark: Them, a rural Danish community, and Copenhagen. Fat consumption was found to be higher in Them, whereas alcohol consumption was higher in Copenhagen. The absolute daily intake of dietary fiber was higher in Them than in Copenhagen. These observations document changes due to industrialization of food production, modern distribution, and marketing methods. PMID- 6296797 TI - Nonstarch polysaccharide consumption in four Scandinavian populations. AB - Nonstarch polysaccharide (NSP) intake was measured in representative samples of 30 men aged 50-59 in 2 urban and 2 rural Scandinavian populations that exhibited a 3-4 fold difference in incidence of large bowel cancer. Intake was measured by chemical analysis of complete duplicate portions of all food eaten over one day by each individual. NSP intakes showed a rural-urban gradient, with 18.4 +/- 7.8 g/day in rural Finland and 18.0 +/- 6.4 g/day in rural Denmark versus 14.5 +/- 5.4 g/day in urban Finland and 13.2 +/- 4.8 g/day in urban Denmark. NSP intakes were also calculated (using food tables) from weighed food records kept over 4 days, one of which was the day on which the duplicate collection was made. Intakes were 2-2.5 g/day higher with this method than with direct chemical analysis, mainly because published tables of values have become outdated and inaccurate as a result of improved methods for measuring NSP in food. Individual variation from day to day in NSP intake was considerable. Average NSP intake and intake of some of its component sugars were inversely related to colon cancer incidence in this geographical comparison. To show a relationship at the individual level between diet and cancer risk in a prospective study would require detailed and accurate methods for the assessment of NSP consumption. PMID- 6296799 TI - Clinical forum. 12. Whooping cough. PMID- 6296798 TI - Colon cancer and large bowel function in Denmark and Finland. AB - Stool weight and transit time through the gut were measured in 4 groups of 30 men, aged 50-59 years, randomly selected from populations in urban (Copenhagen) and rural (Them) Denmark and urban (Helsinki) and rural (Parikkala) Finland. These populations exhibited a 3-4 fold difference in risk for large bowel cancer. Mean transit time (37 +/- 1 hours, Copenhagen; 43 +/- 1 hours, Helsinki; 40 +/- 1 hours, Them; 37 +/- 1 hours, Parikkala) was not significantly different among populations, but average 24-hour stool weights (136 +/- 13 g, Copenhagen; 176 +/- 17 g, Helsinki; 169 +/- 16 g, Them; 196 +/- 15 g, Parikkala) were different and had a significant inverse relationship to total large bowel cancer incidence, with larger stool weights being found in the low-risk population. A high proportion of study subjects, especially in Finland, were found to be taking medication or to have a history of gastrointestinal illness, but neither of these variables related to bowel habit. PMID- 6296800 TI - Anytown anecdotes: a case of crossed lines. PMID- 6296801 TI - Interview with Philip A. Brunell: contagion and varicella-zoster virus. PMID- 6296802 TI - Benign fibrous histiocytoma of the skin. A morphologic study of 290 cases. AB - Two hundred and ninety cases of benign fibrous histiocytoma of the skin are reviewed. Their frequency, distribution according to age, sex and sites, and the architectural and cytological features of the tumors are analyzed. Ninety two cases (31,7%) were male and 198 cases (68,3%) were female. The tumors were most frequently found between second and fourth decades and located mainly in the extremities. Twenty three cases were clinically diagnosed as malignant tumors. Five cytological patterns of benign fibrous histiocytoma of the skin are described: fibrous, vascular, xanthomatous, pleomorphic and giant cell histiocytoma. Their probable histogenesis and the spectrum of morphological variants are discussed. PMID- 6296803 TI - Combined long-term treatment with an LHRH agonist and a pure antiandrogen blocks androgenic influence in the rat. AB - Daily administration for 5 months of the potent LHRH agonist (D-Ser(TBU)6, des Gly-NH2(10)) LHRH ethylamide (250 ng) in combination with the pure antiandrogen RU23908 (5 mg) to adult male rats causes a marked inhibition of ventral prostate and seminal vesicle weight to 9% and 15% of control, respectively. At the doses used, owing to readjustments of the pituitary-testicular axis, neither treatment alone has an effect on prostate weight and exerts only minimal inhibitory effects on seminal vesicle weight. Whereas treatment with the LHRH agonist alone markedly inhibits testicular LH and PRL receptor levels, the antiandrogen alone stimulates the concentration of the two receptors and reverses the inhibitory effect of the LHRH agonist treatment on LH receptors. Treatment with the LHRH agonist decreases plasma PRL levels, whereas the antiandrogen increases the concentration of circulating LH and FSH by 250%. Treatment with the LHRH agonist decreases the concentration of testosterone and its precursors of the delta 4-pathway while stimulating 5 alpha-reductase activity in both the absence and presence of simultaneous treatment with the antiandrogen. The present data show that blockage of the delta 4-steroidogenic pathway induced by treatment with an LHRH agonist prevents the escape phenomenon observed during long-term treatment with a pure antiandrogen, and permits maximal inhibitory effects of the two treatments on secondary sex organ weight. Such combined treatment with an LHRH agonist (to block androgen formation) and an antiandrogen (to neutralize remaining androgens of testicular and adrenal origin) should be the hormonal therapy of choice in prostatic carcinoma. PMID- 6296804 TI - Vulvar disease. A spectrum of clinical pictures. PMID- 6296805 TI - Dietary fiber. Can it prevent certain colonic diseases? AB - Most of the serious organic diseases of the colon are etiologically linked to the high-saturated-fat and low-fiber Western diet. Benign but common conditions, such as appendicitis and diverticular disease of the colon, appear to be due to deficiency of fiber and attendant low-bulk stools. Colon cancer appears to be due to carcinogens created in the colon itself. Contributing to carcinogen production are cocarcinogens in bile and an increase in anaerobic bacteria, both directly related to high levels of saturated fat in the diet. If these common disorders of the colon are to be controlled, our diet will require major modification. Changes will have to include reduction of saturated fats of animal origin and increase in cereal grains. PMID- 6296806 TI - [Clinical significance of the angiotensin-converting enzyme in the serum in sarcoidosis]. PMID- 6296807 TI - [Prophylactic antibiotherapy in heart surgery with cefotiam, clinical evaluation. Non comparative study]. PMID- 6296808 TI - [Immunohistochemical presentation of hormone producing cells in the pancreatic tissue of patients with hyperinsulinism]. PMID- 6296809 TI - [Pathogenesis of the arterial hypertensive syndrome in Itsenko-Cushing disease]. AB - The indices of central hemodynamics and renin-angiotensine-aldosterone system, as well as their relationship, were studied in patients with aggravated Icenko Cushing's disease. A stable high arterial pressure, a raised total peripheral resistance, a decreased cardiac index, an increased blood corticotrophin, cortisol and aldosterone levels and an enhanced plasmatic renin activity were revealed. It is suggested that activation of the renin-angiotensin-aldosterone system stimulates the development of arterial hypertension in patients with Icenko-Cushing's disease, taking into account a significant role of this system in alteration of the central hemodynamics indices. PMID- 6296810 TI - [Preoperative localization of insulinomas]. PMID- 6296812 TI - [Intracellular metabolic characteristics of the megakaryocyte cells of healthy persons]. PMID- 6296811 TI - [Role of cAMP-dependent phosphorylation of rat uterus nuclear proteins in the mechanism of estradiol action]. AB - In experiments on ovariectomized rats cAMP-dependent phosphorylation of the uterine nuclear tissue proteins under the action of estradiol, histamine and cAMP was studied. It was shown, that phosphorylation of the uterine isolated nuclear proteins under the influence of endogenous proteinkinases increases 6 hours after estradiol, histamine and/or cAMP injection to the animals. Proteinkinase activity of the uterine tissue nuclei are higher, when exogenous histone H1 is used as a substrate. The ability of histone H1, isolated from the uterine tissue 6 hours after estradiol, histamine or cAMP injection, to be phosphorylated in vitro with exogenous cAMP-dependent proteinkinase is lowered, that is likely to be resultant of preceding phosphorylation of these proteins in vivo. PMID- 6296813 TI - [Combined cancer and tuberculosis lesions of the lungs]. PMID- 6296814 TI - Effects of skeletal muscle protein phosphatase inhibitor-2 on protein synthesis and protein phosphorylation in rabbit reticulocyte lysates. AB - Reticulocyte lysates contain two major classes of protein phosphatase activities, designated type 1 and type 2. These designations are based on criteria derived from the analyses of protein phosphatase species in other tissues. The criteria include (i) chromatographic elution profiles on DEAE-cellulose; (ii) specificity of lysate phosphatases toward [(32)P]phosphorylase a and [(32)P]phosphorylase kinase; (iii) sensitivity of lysate phosphatases to Mg(2+) ATP; and (iv) sensitivity to the heat-stable protein phosphatase inhibitor-2. The lysate phosphatase species are similar to those described in rabbit skeletal muscle and rabbit liver. Reticulocyte protein phosphatase type 1, but not type 2, is inhibited by heat-stable protein phosphatase inhibitor-1 and -2 which have been characterized from rabbit skeletal muscle. We have initiated a study on the function and specificity of lysate protein phosphatase activities involved in the regulation of protein synthesis by examining the effects of protein phosphatase inhibitor-2 on reticulocyte protein synthesis and protein phosphorylation. Our findings are as follows. (a) Protein phosphatase inhibitor-2 inhibits protein chain initiation in hemin-supplemented lysates. (b) Inhibition is characterized by biphasic kinetics and is reversed by the delayed addition of purified reticulocyte eukaryotic initiation factor 2 (eIF-2). (c) Inhibition of protein synthesis by inhibitor-2 is accompanied by the phosphorylation of the alpha subunit (38,000 daltons) of eIF-2 (eIF-2alpha) and of two heat-stable polypeptides of 29,000 and 44,000 daltons. (d) The 29,000-dalton component is phosphorylated in lysates under conditions of protein synthesis and appears to be inhibitor-2, but the physiological significance of this modification of inhibitor 2 is not clear. (e) Inhibitor-2 has no effect on the activation in vitro of isolated heme-regulated or double-stranded RNA-dependent eIF-2alpha kinases. We propose that the inhibition of protein synthesis in hemin-supplemented lysates by added inhibitor-2 is due at least in part to the inhibition of a type 1 eIF 2alpha phosphatase activity, which permits a basal eIF-2alpha kinase activity to be expressed leading to the accumulation of phosphorylated eIF-2alpha and an inhibition of protein synthesis. PMID- 6296815 TI - Bacteriophage T4-induced anticodon-loop nuclease detected in a host strain restrictive to RNA ligase mutants. AB - The fate of host tRNAs during T4 bacteriophage infection was investigated with Escherichia coli CTr5x, the only known host strain that is restrictive to RNA ligase and polynucleotide kinase mutants. Three CTr5x tRNA species were cleaved during infection. One was leucine tRNA1, which was cleaved in the extra arm, as reported elsewhere for E. coli B infected with bacteriophage T2 or T4. The other two were specific to E. coli CTr5x and were not cleaved in various other hosts. One of the cleaved CTr5x-specific tRNAs had an anticodon sequence of the E. coli B "major" isoleucine tRNA but otherwise little sequence homology. Both CTr5x specific tRNAs were cleaved by a distinct T4-induced endonuclease, other than that of leucine tRNA1, because the CTr5x-specific cleavages (i) were induced later in infection, (ii) persisted with a T4 mutant deficient in leucine tRNA1 endonuclease, and (iii) occurred in the anticodon loop. The specific manifestation of the anticodon-directed endonuclease activity in T4-infected E. coli CTr5x suggests roles for RNA ligase and polynucleotide kinase in processing of host tRNA species. PMID- 6296816 TI - In vitro synthesis and integration into mitochondria of porin, a major protein of the outer mitochondrial membrane of Saccharomyces cerevisiae. AB - We have isolated an outer mitochondrial membrane (OMM) fraction from baker's yeast. Saccharomyces cerevisiae, that possesses porin activity and contains a major polypeptide of 29,000 daltons. By analogy to similar data for an OMM fraction from rat liver and mung bean [Zalman, L. S., Nikaido, N. & Kagawa, Y. (1980) J. Biol. Chem. 255, 1771-1774], the 29,000-dalton polypeptide of the isolated yeast OMM fraction has been tentatively identified as porin. Evidence to substantiate this identification was provided by the finding that both the porin activity and the 29,000-dalton polypeptide were entirely resistant when the OMM fraction was exposed to trypsin digestion, with the 29,000-dalton polypeptide being virtually the only polypeptide in the OMM fraction to be unaffected by trypsin digestion. There was no protection when trypsin digestion was carried out in the presence of detergent. Using monospecific antibodies, we have shown that yeast porin is apparently not synthesized as a larger precursor in a cell-free translation system. In vitro-synthesized porin could not be integrated into dog pancreas microsomal vesicles or into an isolated OMM fraction from yeast, either co- or posttranslationally. In vitro-synthesized porin, however, could be integrated posttranslationally into whole isolated mitochondria. This membrane specificity suggests that integration does not proceed by unassisted partitioning. The integration of porin into whole mitochondria occurred with fidelity by the criterion of its resistance to trypsin. Moreover, integration was not inhibited in the presence of the protonophore carbonyl cyanide m-chlorophenyl hydrazone whereas translocation into the mitochondrial matrix of the in vitro synthesized gamma subunit of F1-ATPase was inhibited. PMID- 6296817 TI - Transit of pp60v-src to the plasma membrane. AB - The protein kinase (pp60v-src) encoded by the transforming gene (v-src) of Rous sarcoma virus is synthesized on free polyribosomes and then translocated to the plasma membrane of infected cells. Neither the mechanism of the translocation nor the physiological significance of the membrane localization has been elucidated. We have explored these problems by pursuing previous observations of a complex between pp60v-src and two cellular proteins with molecular weights of 50,000 and 89,000. We found the complex located entirely in the cytoplasm, where it appears to form immediately after the synthesis of pp60v-src. While in the complex, pp60v src has little detectable kinase activity and is phosphorylated predominantly on serine. After transfer from the complex to the plasma membrane, pp60v-src becomes phosphorylated on tyrosine as well as serine and acquires kinase activity. Under restrictive conditions, temperature-sensitive pp60v-src is produced in normal quantities, but translocation to the plasma membrane is diminished. As an apparent consequence, the cytoplasmic complex accumulates to abnormal abundance. Alternatively, temperature-sensitive pp60v-src that has been synthesized and translocated to the plasma membrane under permissive conditions appears to be released from the membrane and returns to the cytoplasmic complex when the infected cells are shifted to the restrictive temperature. We conclude that the cytoplasmic complex may be the vehicle by which pp60v-src reaches the plasma membrane. It is possible that other proteins may follow a similar route to the membrane. Binding to plasma membrane appears to be a discrete step in the biogenesis of pp60v-src and may be essential to the function of the protein. PMID- 6296818 TI - Purified matrix protein of vesicular stomatitis virus blocks viral transcription in vitro. AB - One of the major structural proteins of vesicular stomatitis virus is a small, nonglycosylated, matrix protein which associates with the nucleocapsid core during final stages of morphogenesis and budding. Biochemical and genetic studies suggested that the matrix protein regulates RNA synthesis both in vitro and in vivo. We have purified biologically active matrix protein from the virus and have directly shown that it significantly inhibits RNA synthesis in vitro mediated by the virion-associated RNA polymerase at low ionic strength (0.02 M). The inhibition was greater than 80% when the ratio of matrix protein to the major nucleocapsid protein in the transcribing complex was 2:1 (wt/wt). The inhibition was found to be at the level of RNA chain elongation and not at the initiation step. Electron microscopic studies revealed that inhibition of transcription by matrix protein was accompanied by a profound structural change of the transcribing nucleocapsid from an extended structure to a highly compact form. At higher ionic strength (0.12 M), the matrix protein failed to interact with the nucleocapsid. The matrix protein appears to be involved in condensing the nucleocapsid and blocking transcription during maturation of the virus particle. PMID- 6296819 TI - Changes in the topography of early region transcription during polyoma virus lytic infection. AB - We have studied the pattern of transcription of the early region of polyoma virus DNA after the onset of the late phase of lytic infection of mouse cells. Following initiation of viral DNA synthesis, the early/late switch is accompanied not only by efficient production of late mRNAs but also by the appearance of previously unidentified early-strand RNAs which have certain structural features in common with the classical early mRNAs. Stable poly(A)+RNAs have been identified by blot analysis and S1 nuclease mapping that are not detected early during infection or in polyoma virus-transformed cells. One group consists of transcripts whose 5' ends map 150-200 nucleotides upstream from the major early 5' ends (at positions 148 and 153 on the polyoma virus genome) but whose splicing pattern and poly(A) addition sites are indistinguishable from those of mRNAs produced early in infection. The 5' exons of these early region transcripts contain an open translational reading frame that extends from nucleotide positions 5,255 to 124 and is capable of encoding a basic protein of 53 amino acids. Transcription of these RNAs does not appear to be negatively regulated by large tumor antigen. A transcript of 1,800 nucleotides appears to map predominantly between 93 and 26 map units and does not contain sequences present in the early mRNA 5' exons. These data suggest that, after the onset of polyoma DNA replication, the activation of new early-strand promoters leads to the expression of previously untranscribed viral DNA sequences. PMID- 6296820 TI - Transformation and replication in mouse cells of a bovine papillomavirus--pML2 plasmid vector that can be rescued in bacteria. AB - The unique ability of bovine papillomavirus (BPV-1) DNA to replicate as a stable, multicopy plasmid in transformed mouse cells has led to its utilization as a eukaryotic cloning vector. One limitation of the system has been the marked reduction in transformation efficiency when BPV-1 DNA or the subgenomic transforming segment of BPV-1 DNA (BPV69T) is covalently linked to pBR322 sequences. A dual host replicon consisting of BPV-1 DNA and pML2d, a deletion variant of pBR322, was constructed and shown to be highly efficient for transformation of mouse cells in vitro. The hybrid molecule replicates as a stable, unintegrated, multicopy plasmid in transformed mouse cells. The resident BPV-1-pML2d plasmid DNA was rescued in bacteria and the recovered plasmids were shown to be identical in structure and to have the same transformation efficiency as the original transforming DNA. In contrast, the transforming efficiency of BPV69T DNA is less than 1/100th that of BPV-1 DNA when the DNA is left covalently linked to pML2d. These observations indicate that, although the nontransforming region of BPV-1 (BPV31NT) DNA is not essential for transformation, it has a facilitative role in the transformation process. PMID- 6296821 TI - Complete amino acid sequence of pig kidney fructose-1,6-bisphosphatase. AB - The covalent structure of the pig kidney fructose-1,6-bisphosphatase (D-fructose 1,6-bisphosphate 1-phosphohydrolase, EC 3.1.3.11) subunit has been determined. Placement of the 335 amino acid residues in the polypeptide chain was based largely on automated Edman degradation of eight purified cyanogen bromide fragments generated from the S-carboxymethylated protein. The determination of the amino acid sequence of the largest cyanogen bromide fragment (154 residues) required additional analysis of subfragments obtained by tryptic cleavage at arginyl residues and by mild acid cleavage of an Asp-Pro peptide bond. Alignment of the cyanogen bromide fragments was accomplished by analysis of a product of limited proteolysis by an endogenous protease and by characterization of the tryptic peptides isolated from S-[14C]carboxymethylated fructose-1,6 bisphosphatase. This sequence information has permitted the identification of several reactive sites of functional and structural significance in pig kidney fructose-1,6-bisphosphatase. PMID- 6296822 TI - Crystallization of mitochondrial cytochrome oxidase. AB - Cytochrome oxidase (ferrocytochrome c:oxygen oxidoreductase, EC 1.9.3.1) was purified from beef heart mitochondria. By washing the oxidase with detergent on a hydrophobic interaction column, phospholipids were depleted to the level of 1 mol of cardiolipin per mol of heme a. Hydrophobic impurities and partially denatured oxidase were separated from the intact oxidase on an affinity column with cytochrome c as the specific ligand. The final preparation of the oxidase contained seven distinct polypeptides. The molecular weight of the oxidase was estimated to be 130,000 from its specific heme a and copper content and from the subunit composition. Crystals of the oxidase were obtained by slow removal of the detergent from the buffer in which the oxidase was dissolved. The needle-shaped crystals were 100 microns in average length and 5 microns in width, and they strongly polarized visible light. Electron diffraction patterns were obtained with an unstained glutaraldehyde-fixed single crystal by electron microscopy using 1,000-kV electrons. From electron micrographs and the diffraction patterns of the crystal, it was concluded that the crystal is monoclinic in the space group P21, with unit cell dimensions a = 92 A, b = 84 A, and c = 103 A, and alpha = beta = 90 degrees, gamma = 126 degrees. PMID- 6296823 TI - Conformational implications of enzymatic proline hydroxylation in collagen. AB - In 1979 it was proposed that prolyl hydroxylase (prolyl-glycyl-peptide,2 oxoglutarate:oxygen oxidoreductase, EC 1.14.11.2) recognizes the beta-turn conformation in nascent procollagen chains and that the hydroxylation process involves a conformational change resulting in "straightening" of the beta-turn segments into the linear triple-helical conformation of native collagen. We present experimental data that verify both these postulates. The following peptides were synthesized and studied for their conformation and interaction with prolyl hydroxylase: tBoc-Pro-Gly-Ala-OH, tBoc-Pro-Gly-Val-OH, tBoc-Gly-Val-Pro Gly-Val-OH, and tBoc-Pro-DAla-Ala-OH. Spectral data showed that these peptides preferred a beta-turn conformation. All of them acted as inhibitors of the enzyme; the pentapeptide also acted as a substrate. To mimic the biosynthetic event, a collagen model polypeptide, (Pro-Pro-Gly)10, was incubated at 37 degrees C with purified prolyl hydroxylase and the necessary cosubstrates and cofactors at pH 7.8. A progressive change from the initially nonhelical to the triple helical conformation, as monitored by CD spectra and gel filtration, occurred during the course of proline hydroxylation. In addition to leading to increased thermal stability of the triple-helical conformation in (Pro-Pro-Gly)10 and (Pro Pro-Gly)5, the enzymatic incorporation of the hydroxyproline residues was found to enable these polypeptides to fold into this conformation faster than the unhydroxylated counterparts. These conformational implications of proline hydroxylation in collagen may also be of use in the study of the complement subcomponent Clq and of acetylcholine esterase which contain collagen-like regions in them. PMID- 6296824 TI - Proton translocation stoichiometry of cytochrome oxidase: use of a fast responding oxygen electrode. AB - The mechanistic stoichiometry of vectorial H+ ejection coupled to electron transport from added ferrocytochrome c to oxygen by the cytochrome oxidase (EC 1.9.3.1) of rat liver mitoplasts was determined from measurements of the initial rates of electron flow and H+ ejection in the presence of K+ (with valinomycin). Three different methods of measuring electron flow were used: (a) dual-wavelength spectrophotometry of ferrocytochrome c oxidation, (b) uptake of scalar H+ for the reduction of O2 in the presence of a protonophore, and (c) a fast-responding membraneless oxygen electrode. The reliability of the rate measurements was first established against the known stoichiometry of the scalar reaction of cytochrome oxidase (2ferrocytochrome c + 2H+ + 1/2O2 leads to 2ferricytochrome c + H2O) in the presence of excess protonophore. With all three methods the directly observed vectorial H+/O ejection ratios in the presence of K+ + valinomycin significantly exceeded 3.0. However, because the rate of backflow of the ejected H+ into the mitoplasts is very high and increases with the increasing delta pH generated across the membrane, there is a very rapid decline in the observed H+/O ratio from the beginning of the reaction. Kinetic analysis of ferrocytochrome c oxidation by the mitoplasts, carried out with a fast-responding membraneless oxygen electrode, showed the reaction to be first order in O2 and allowed accurate extrapolation of the rates of O2 uptake and H+ ejection to zero time. At this point, at which there is zero delta pH across the membrane, the H+/O ejection ratio of the cytochrome oxidase reaction, obtained from the rates at zero time, is close to 4.0. PMID- 6296825 TI - Functional reconstitution of beta-adrenergic receptors and the stimulatory GTP binding protein of adenylate cyclase. AB - A procedure for the functional reconstitution of beta-adrenergic receptors and the stimulatory guanine nucleotide-binding protein (G/F) of adenylate cyclase in phospholipid vesicles is described. beta-Adrenergic receptors were solubilized from turkey erythrocyte plasma membranes and reconstituted into phospholipid vesicles by the addition of dimyristoyl phosphatidylcholine and removal of detergent by gel filtration. This procedure restored the ability to bind [125I]iodohydroxybenzylpindolol and [3H]dihydroalprenolol. Purified rabbit hepatic G/F that was added to the receptor vesicles could be stably activated by guanosine 5'-[3-thio]triphosphate at a low rate, and this activation was increased up to 4-fold in the presence of beta-adrenergic agonists. This stimulation of the activation of G/F was specific for beta-adrenergic agonists and could be specifically blocked by beta-adrenergic antagonists. Stimulation was proportional to the concentration of vesicles containing active beta-adrenergic receptor. Under optimal conditions, 5 to 6 molecules of G/F were activated per receptor, indicating that catalytic activation of G/F by receptor was reconstituted. PMID- 6296826 TI - A transposable element from Halobacterium halobium which inactivates the bacteriorhodopsin gene. AB - We describe the characterization of a transposable element from an archaebacterium. The bacteriorhodopsin genes from the wild-type and two mutant Halobacterium halobium strains have been cloned as BamHI fragments in pBR322. The cloned DNA fragments from the two mutants both contain a 1.1-kilobase-pair insertion sequence (ISH1) near the NH2 terminus of the bacteriorhodopsin coding sequence. ISH1 is present in the two mutants in an identical palindromic site but in opposite orientations. The complete sequence of ISH1 has been determined; it is 1,118 nucleotides long, it has 8-base-pair interrupted inverted repeats at the ends, and it duplicates an 8-base-pair (A-G-T-T-A-T-T-G) target sequence upon insertion. As for most eukaryotic and some prokaryotic transposable elements, the sequence of the ISH1 begins with T-G and ends in C-A. ISH1 contains an open reading frame 810 nucleotides long and codes for an RNA approximately 900 nucleotides long. The copy number of ISH1 ranges from one to five or more in different H. halobium strains. In at least one of the strains, one copy of ISH1 is present also on a plasmid DNA. PMID- 6296827 TI - Electron paramagnetic resonance crystallography of bacterial catalase: g-Contour mapping method of analysis. AB - Single crystals of bacterial catalase from Micrococcus luteus have been examined by EPR at 77 K. X-ray perfect crystals gave a set of four prominent resonances in all three orthogonal planes which yielded eight heme direction cosine matrices to an accuracy of +/- 2 degrees as expected for the P4(2)2(1)2 space group and unit cell parameters previously determined. These matrices are related by D4 symmetry operation of the space group. There were additional weaker resonances only resolved in two or even one plane. A method of g-contour mapping was devised to solve for the orientations of hemes that give rise to these weaker resonances. Three additional sets of heme orientations, also following D4 symmetry, were determined. All of the above sites have the same principal g values, 2.0, 5.4, and 6.6. The EPR crystallographic results imply that several conformational substates may be trapped at 77 K. PMID- 6296828 TI - Gonadotropin-releasing hormone stimulation of luteinizing hormone release: A ligand-receptor-effector model. AB - A divalent antibody conjugate of a pure antagonist of gonadotropin-releasing hormone (GnRH) behaved as an agonist--i.e., released luteinizing hormone (LH) from pituitary cultures. Release was measured over a wide range of conjugate concentrations; it rose to a maximum of 66% of the LH released by the optimal concentration of GnRH and declined to basal levels at very high concentrations. This behavior was modeled on the assumption that the antibody conjugate, A, can react with a receptor, R, to form a complex, A . R2. This dimer then can react with a quiescent effector, E (e.g., a closed Ca2+ ion channel), to form A . R2 . E, which contains activated effector and leads to cellular responses. The equilibrium equations governing the behavior of this model were derived, solved, and found to yield a good fit to the experimental data. Consideration of our data in this model system, and of other available data describing the behavior of ligands in other cells, suggests that the present model may be of wide applicability. PMID- 6296829 TI - Polyadenylylated RNA complementary to a mouse retrovirus-like multigene family is rapidly and specifically induced by epidermal growth factor stimulation of quiescent cells. AB - Complementary DNA probes prepared from total polysomal poly(A)+RNA populations were used to identify clones of mouse DNA containing sequences whose expression is specifically enhanced after epidermal growth factor (EGF) stimulation of quiescent mouse embryo cells in culture. Three such clones were isolated and used to study changes in the levels of clone-specific poly(A)+RNA in the polysomes of cells after mitogenic stimulation by EGF. RNA complementary to sequences present in these clones increased approximately equal to 10-fold as a fraction of the total poly(A)+RNA by 6 hr after stimulation. All three clones were found by hybridization criteria to contain sequences related to the class of mouse retrovirus or transposon-like elements termed VL30. These VL30-related sequences were further found to be complementary to EGF-inducible poly(A)+RNAs and enhanced expression was detectable as early as 1 hr after EGF stimulation. In contrast, nine additional clones, including an AKR-type murine leukemia provirus DNA clone, contained no detectable VL30 sequence elements and were complementary to poly(A)+RNA species whose relative concentration was essentially constant in quiescent and EGF-stimulated cells. Therefore, VL30 sequence elements appear distinct in that they encompass members whose expression is specifically regulated in response to a defined peptide growth factor. PMID- 6296830 TI - A DNA-mediated transformation system for Dictyostelium discoideum. AB - We have established a transformation system for Dictyostelium discoideum. The transformation vector contains the protein coding region of the Tn5 neomycin resistance gene fused to the proposed promoter of the Dictyostelium actin 8 gene; the vector also contains a sequence that acts as an autonomously replicating sequence (ars) in yeast. Using this vector, we can transform Dictyostelium vegetative amoebae to be resistant to aminoglycoside G418 at a frequency of between 10(-6) and 10(-4) of the input cells. The transformed cell lines are stable and contain vector sequences integrated within chromosomal DNA. PMID- 6296831 TI - Vaccinia virus: a selectable eukaryotic cloning and expression vector. AB - Foreign DNA was inserted into two nonessential regions of the vaccinia virus genome by homologous recombination in cells infected with virus and transfected with plasmids containing the foreign DNA elements flanked by vaccinia virus DNA. Thymidine kinase-negative (TK-) recombinants were selected after inserting foreign DNA into the coding region of the TK gene of wild-type vaccinia virus; TK+ recombinants were selected after inserting the herpesvirus TK gene into TK- mutants of vaccinia virus. For TK+ expression, it was necessary to insert a 275 base-pair DNA fragment containing the initiation site and sequences upstream of an early vaccinia virus transcript next to the coding sequences of the herpesvirus gene. The unique ability of the herpesvirus TK to phosphorylate 125I labeled deoxycytidine provided independent confirmation of gene expression. These studies demonstrate the use of vaccinia virus as a selectable cloning and expression vector, confirm the map location of the vaccinia virus TK gene, and provide initial information regarding the location of vaccinia virus transcriptional regulatory sequences. PMID- 6296832 TI - Mutant immunoglobulin genes have repetitive DNA elements inserted into their intervening sequences. AB - The kappa light chain genes from two mutant hybridoma cell lines defective in kappa light chain synthesis were isolated and compared to the wild-type kappa light chain gene. In each case, the mutant kappa light chain genes were found to contain repetitive DNA elements in their intervening sequences that were not present in the intervening sequences of the wild-type kappa light chain gene. These elements were found to be related to the genes of intracisternal A particles. These results suggest that the decreased production of kappa light chain in the mutant cell lines is due to the presence of the intracisternal A particle-related genes. PMID- 6296833 TI - A transposable element that splits the promoter region inactivates a Drosophila cuticle protein gene. AB - Two mutations that affect larval cuticle protein gene expression in the 2/3 variant Drosophila melanogaster strain were investigated. We demonstrate that this strain synthesizes an electrophoretic variant, fast 2 (CPf2), of wild-type cuticle protein 2(CP2). It also lacks detectable amounts of cuticle protein 3 (CP3). The other major cuticle proteins are still present. Protein and DNA sequence analyses indicate that point mutations cause two amino acid substitutions that change the electrophoretic mobility of CPf2 relative to that of CP2. The mutation abolishing the expression of CP3 was found to be a 7.3 kilobase DNA insertion located within the T-A-T-A box region of this gene, at -31 base pairs from the mRNA start site. This DNA insertion, called H.M.S. Beagle, belongs to a conserved family of repeated DNA elements that have characteristics similar to those of previously characterized Drosophila transposable elements. H.M.S. Beagle elements are repeated approximately 50 times in the haploid genome and exhibit restriction fragment-length polymorphisms around points of insertion between Canton S, Oregon R, and 2/3 Drosophila strains. Sequence analysis indicates that H.M.S. Beagle contains 266-base-pair direct repeats at its termini and is flanked by a duplication of 4 base pairs of target DNA sequence, T-A-T-A, in the CP3 gene insertion. Thus, insertion of a transposable element into the putative promoter region of the CP3 gene is evidently responsible for inactivating CP3 gene expression. PMID- 6296834 TI - Control of transposon Tn5 transposition in Escherichia coli. AB - Tn5 is a composite transposable element in which the insertion sequences IS50R and IS50L bracket a central region encoding kanamycin resistance (kanr). IS50R encodes a functional transposase, whereas IS50L contains the promoter of the kanr gene. To determine the relative activities of IS50R and IS50L in transposition we examined the structures of chimeric DNA molecules generated by insertion of segments of pBR322::Tn5 dimeric plasmids into red- lambda phage in recA- Escherichia coli. Restriction endonuclease analyses showed that the inserted sequences contained direct terminal repeats of pairs of IS50R or of IS50L elements and that the frequencies of usage of IS50R vs. IS50L depended on the position and orientation of Tn5 in the plasmid vector: IS50R was used preferentially when Tn5 was in transcriptionally quiescent regions of the vector (in either orientation) or when IS50L was immediately downstream from a strong promoter in the vector. In contrast, IS50L was used preferentially when IS50R was downstream from a strong promoter. We conclude IS50R tends to be used preferentially but that when transcription impinges on the end of an IS50 element the participation of that element in transposition is inhibited. PMID- 6296835 TI - Spasmogenic activity of chemotactic N-formylated oligopeptides: identity of structure--function relationships for chemotactic and spasmogenic activities. AB - The chemotactic N-formylated oligopeptides are potent spasmogenic agents for guinea pig ileum. Structure-activity studies with various N-formylated peptides suggest the presence of a specific receptor that resembles in specificity the formyl peptide receptor on leukocytes. A competitive antagonist of the formyl peptide receptor on leukocytes also inhibits formyl peptide-induced ileum contraction, whereas the antihistamine diphenhydramine is without effect. The contractile response caused by the synthetic N-formylated peptides differs from those induced by acetylcholine, histamine, and substance P. In particular, a latent period after treatment with the N-formyl peptides is seen before the onset of the response, and a sustained contractile response is not maintained. In addition, tachyphylaxis does occur, but complete recovery of activity is seen after a 20- to 30-min rest period. These observations suggest broad biological roles of prokaryotic signal peptides from bacteria as acute inflammatory mediators. PMID- 6296836 TI - Purified Epstein-Barr virus Mr 340,000 glycoprotein induces potent virus neutralizing antibodies when incorporated in liposomes. AB - The purified Mr 340,000 glycoprotein component of Epstein-Barr (EB) virus-induced membrane antigen complex incorporated into liposomes was shown to be a potent immunogen in mice. High-titer antisera were induced that (i) are specific for membrane antigen components without absorption, (ii) bind the antigens induced by three different EB virus isolates, and (iii) neutralize the ability of the virus to transform fetal cord blood lymphocytes in vitro. The development of this immunogenic form of purified antigen provides an important step towards a potential subunit vaccine against Epstein-Barr virus infection. PMID- 6296837 TI - Isolated poliovirus capsid protein VP1 induces a neutralizing response in rats. AB - Antibodies were raised in rats against the poliovirus type 1 capsid proteins, VP1, VP2, and VP3. Antibodies directed against VP1 from type 1 poliovirus (Mahoney) neutralized type 1 but not type 2 poliovirus. Antibodies raised against VP2 and VP3 failed to neutralize type 1 virus. Thus, VP1 appears to be a neutralizing antigen for poliovirus and in its denatured form presents to the immune system its neutralizing determinants. PMID- 6296838 TI - Abnormal adenosine-induced immunosuppression and cAMP metabolism in T lymphocytes of patients with systemic lupus erythematosus. AB - Normal human T lymphocytes incubated with adenosine (10 muM) for 30 min at 37 degrees C show an increase in the percentage of cells expressing receptors for the Fc portion of IgG (RFc(gamma)) and the OKT8 antigen, while the proportion of OKT4(+) cells decreases. These effects occur exclusively in a subset of T cells with theophylline-resistant sheep erythrocyte receptors (T(R) cells) that is enriched for OKT4(+) cells. Untreated normal T(R) cells express helper/inducer cell activity for T-cell-dependent B-cell differentiation, while adenosine treated T(R) cells suppress B-cell differentiation. In contrast, in T(R) cells isolated from patients with systemic lupus erythematosus (SLE), adenosine fails to induce immunosuppressor activity or to increase the percentage of OKT8(+) and RFc(gamma) (+) cells. In addition, although incubation of normal T(R) cells with adenosine causes a transient increase in cAMP levels (up to 160% of control within 5 min), in SLE T(R) cells, cAMP levels fall by 50% within 10 min. The photoaffinity label 8-azidoadenosine cyclic [(32)P]monophosphate has been used to show that human T lymphocytes have a single cAMP receptor site that appears to be the regulatory subunit of type I protein kinase. In normal T(R) cells, this receptor becomes occupied in response to adenosine. In contrast, in SLE T(R) cells, no change in cAMP receptor occupancy is detected. Although adenosine has a differential effect on normal and SLE T(R) cells, cAMP derivatives that can traverse the cell membrane (8-bromo- and 8-azidoadenosine cyclic monophosphates) induce an increase in the RFc(gamma) (+) cell subset in both normal and SLE T(R) cells. These results suggest that cAMP mediates the effects of adenosine on cell surface markers of T lymphocytes. The lack of an adenosine receptor-coupled adenylate cyclase activity in SLE T(R) cells may account, in part, for their lack of immunosuppressive activity. PMID- 6296839 TI - Prostaglandin biosynthesis in the human fetal adrenal gland: regulation by glucocorticosteroids. AB - Human fetal adrenal (HFA) tissue was maintained in organ culture to evaluate the biosynthesis of prostaglandins and hormonal regulation of prostaglandin formation by this tissue. The HFA tissue secreted substantial amounts of prostaglandin E(2), prostaglandin F(2alpha), 13,14-dihydro-15-ketoprostaglandin F(2alpha), 6 ketoprostaglandin F(1alpha), and thromboxane B(2); secretion of prostaglandin D(2) could not be demonstrated. Prostaglandin biosynthesis in HFA tissue was inhibited in a time-dependent manner by corticotropin (ACTH; 0.4 muM); by the fourth day of culture, the extent of inhibition of biosynthesis of each prostaglandin was 60-90%. Progesterone (1 muM), cortisol (1 muM), and dexamethasone (1 muM) inhibited prostaglandin biosynthesis whereas estradiol (1 muM) did not. Of the compounds tested for inhibitory activity, dexamethasone was the most potent. An inhibitor of 11beta-hydroxylase activity (metyrapone; 0.1 mM) effectively eliminated the inhibition of prostaglandin biosynthesis caused by corticotropin and progesterone. Metyrapone treatment alone caused a 3-fold increase in prostaglandin biosynthesis by fetal adrenal tissues. Similar stimulatory effects resulted from treatment with inhibitors of (i) 3beta hydroxysteroid dehydrogenase (cyanoketone; 15 muM), (ii) steroid 17alpha hydroxylase (SU 10603; 19 muM), and (iii) cholesterol side-chain cleavage (aminoglutethimide; 1 mM). Inhibition of prostaglandin biosynthesis by dexamethasone in the presence or absence of metyrapone was concentration dependent and 50% inhibition could be demonstrated at 1 nM. A competitive inhibitor of the binding of glucocorticosteroids to cytoplasmic receptors (cortisol 21-mesylate; 1 muM) significantly reduced the inhibition of prostaglandin biosynthesis effected by dexamethasone (10 nM). These findings suggest that prostaglandin biosynthesis in the HFA gland is regulated by endogenously synthesized glucocorticosteroids, the actions of which are mediated by a glucocorticosteroid receptor. Such glucocorticosteroids induce the synthesis of a substance that inhibits prostaglandin biosynthesis. PMID- 6296840 TI - Use of a monoclonal antibody to purify the tetrodotoxin binding component from the electroplax of Electrophorus electricus. AB - The tetrodotoxin binding component of the voltage-sensitive sodium channel from Electrophorus electricus electroplax was purified by using a monoclonal antibody. An impure preparation of tetrodotoxin binding component was mixed with the pure monoclonal antibody, and the immune complex so formed was isolated by affinity chromatography on a protein A-Sepharose column. Excess antibody was removed by ion-exchange chromatography. The purified material has a specific activity of over 1,800 pmol of [3H]tetrodotoxin bound per mg of protein. By assuming that the immune complex has a stoichiometry of 1:1, this specific activity then represents an actual specific activity of 3,000 pmol of [3H]tetrodotoxin bound per mg of eel electroplax protein, or 75% of the theoretical specific activity expected for a pure toxin binding component of Mr 250,000. The peptide composition of the purified material was simple with the predominant species present being of Mr approximately equal to 250,000. Minor components were also present with Mrs of approximately equal to 95,000, approximately equal to 44,000, and approximately equal to 23,000. PMID- 6296841 TI - Opiates and classical conditioning: selective abolition of conditioned responses by activation of opiate receptors within the central nervous system. AB - It has previously been shown that opiates produce selective abolition of aversively motivated classically conditioned responses in the rabbit. The experiments reported here show that these effects are mediated by specific activation of opiate receptors within the central nervous system in that this central activation is both necessary and sufficient to produce opiate-induced abolition of conditioned responding. Further characterization suggests that selective activation through opiate-mu-receptor interactions within the periaqueductal gray/periventricular region of the fourth ventricle may be critical in mediating this abolition. PMID- 6296842 TI - Tetanus toxin fragment forms channels in lipid vesicles at low pH. AB - Single-walled asolectin vesicles loaded with K+ at pH 7.00 released their K+ content upon incubation with tetanus toxin fragment B but only when the incubation was at pH below 5.00. Whole tetanus toxin exhibited only a weak releasing activity. Toxin light alpha chain and the carboxyl-terminal 48,000 dalton moiety of the heavy chain (fragment IIC) were unable to provoke K+ release from vesicles at any pH. K+ release from lipid vesicles could also be detected with tetanus toxin heavy beta chain at low pH. Furthermore, using a detergent binding assay ([3H]Triton X-100), we have also shown that an hydrophobic domain, localized in the 50,000-dalton terminal polypeptide of tetanus toxin heavy chain, is detectable at pH 3.60 but not at pH 5.00. These results lead us to conclude that the ability of tetanus toxin fragment B to release K+ from asolectin vesicles at low pH is due to the 50,000-dalton amino-terminal polypeptide of the heavy chain present in toxin fragment B. We propose that this phenomenon is caused by channel formation across the vesicle membrane as has been observed for the 23,000-dalton amino-terminal moiety of diphtheria toxin fragment B. PMID- 6296843 TI - Identification of a human neuroectodermal tumor antigen (OFA-I-2) as ganglioside GD2. AB - Two monospecific human antibodies (anti-OFA-I-1 and anti-OFA-I-2) produced in vitro by lymphoblast cell lines originating from melanoma patients have been shown previously to recognize cell surface antigens (OFA-I-1 and OFA-I-2) on human tumors and fetal brain: OFA-I-1 is expressed on a variety of human tumors, while OFA-I-2 has been detected only on tumors of neuroectodermal origin. Evidence presented in this report suggests that the two antigens expressed by a cultured human melanoma cell line (M14) are chemically distinct and that OFA-I-2 is a cell surface glycolipid, ganglioside GD2: GalNAc beta 1 leads to 4 NeuAc alpha 2 leads to 8NeuAc alpha 2 leads to 3 Gal beta 1 leads to 4Glc-ceramide. PMID- 6296844 TI - Glucocorticoid and thyroid hormones transcriptionally regulate growth hormone gene expression. AB - In order to define the molecular mechanisms by which glucocorticoids and thyroid hormone act to regulate growth hormone gene expression, the sites at which they exert their effects on growth hormone biosynthesis were examined in vivo and in a pituitary cell line. Glucocorticoids were shown to rapidly increase accumulation of growth hormone mRNA and nuclear RNA precursors. Glucocorticoids and thyroid hormone were shown to rapidly and independently increase growth hormone gene transcription. These events are shown to occur physiologically in animals and further establish the importance of growth hormone gene expression as a model for steroid regulation. PMID- 6296845 TI - The cAMP-binding domains of the regulatory subunit of cAMP-dependent protein kinase and the catabolite gene activator protein are homologous. AB - Comparison of the recently determined amino acid sequences of the regulatory subunit of cAMP-dependent protein kinase (RII) from bovine cardiac muscle and the Escherichia coli catabolite gene activator protein (CAP) shows significant homology. This homology extends over most of the amino-terminal domain in CAP and is particularly good for the region of the beta-roll structure. The RII sequence contains two adjacent and internally homologous regions, both of which have high resemblance to the cAMP-binding domain in CAP. This suggests that the protein kinase regulatory subunit contains two cAMP-binding domains in the carboxyl terminal region, each having a beta-roll structure similar to that in CAP. The cAMP molecule is expected to bind to the RII within a pocket formed by residues from the beta-roll, as is the case with CAP. One cAMP molecule would interact with residues from about 163 to 220, and the other cAMP would interact with amino acids in the stretch 285-350 of the RII protein kinase sequence. As the carboxyl terminal domain of CAP shows homologies to the DNA-binding domains of other regulatory proteins, the protein appears to be of modular construction: a DNA binding domain joined to a cAMP-binding domain. PMID- 6296846 TI - Host range mutants of vesicular stomatitis virus defective in in vitro RNA methylation. AB - The viral RNA polymerase of detergent-treated vesicular stomatitis virus normally synthesizes viral mRNAs in vitro that are both guanylylated and methylated to give 5'-terminal 7mGpppAm caps. We have characterized a virus host range mutant, hr 1, that is totally defective in vitro in the methylation of mRNA, although full-length polyadenylylated mRNAs with 5' termini of the form GpppA are synthesized in normal yields. A second mutant, hr 8, is partially defective in methylation and synthesizes mRNAs in vitro with primarily GpppA and some GpppAm 5' termini. When used for in vitro translation, the unmethylated hr 1 mutant mRNA shows, as expected, reduced synthesis of viral proteins. These data provide direct evidence that the vesicular stomatitis virus-associated methyltransferase activities are virus encoded. PMID- 6296847 TI - Development of hybridomas secreting monoclonal antibodies to the chicken intestinal 1 alpha,25-dihydroxyvitamin D3 receptor. AB - Four hybridomas that secrete monoclonal antibodies to the chicken intestinal cytoplasmic 1 alpha,25-dihydroxyvitamin D3 [1,25-(OH)2D3] receptor have been obtained. Splenic lymphocytes, derived from two male Lewis rats expressing serum antireceptor activity after repeated immunization with a partially purified preparation of this protein, were fused with two nonsecreting murine myeloma cell lines, SP2/0-Ag14 and P3-X63-Ag8.653. Viable hybrids were screened for anti chicken intestinal 1,25-(OH)2D3 receptor activity by incubation of hybrid media with receptor-hormone complex; this was followed by immunoprecipitation with rabbit anti-rat IgG. Of 1,724 hybridomas assayed by this technique, 4 were positive (2 derived from each animal) for the secretion of an antireceptor immunoglobulin molecule. After cloning by limiting dilution, the cell lines (designated SP2/0-4A5, P3-8C8, SP2/0-8D3, and SP2/0-9A7) were expanded into suspension culture. Antibody-induced alterations in the sedimentation pattern of the native 1,25-(OH)2D3 receptor, coupled with Ouchterlony double-diffusion techniques, indicate that SP2/0-4A5 secretes an IgG2a, SP2/0-9A7 produces an IgG2b, and P3-8C8 secretes an IgG. In contrast, SP2/0-8D3 was found to synthesize an IgM. The monoclonal antibodies react with both occupied and unoccupied chicken intestinal receptor and nuclear receptors, and they crossreact with 1,25-(OH)2D3 receptors from a wide variety of tissue and cultured cell types, including human 1,25-(OH)2D3 receptors. These immunological reagents should prove valuable in the elucidation of the molecular action of 1,25-(OH)2D3. PMID- 6296848 TI - Isolation of Drosophila proteins that bind selectively to left-handed Z-DNA. AB - An affinity column for isolating Z-DNA binding proteins was made by attaching brominated poly(dG-dC) to Sephadex. Proteins from Drosophila nuclei were prepared and those that could bind to Escherichia coli B-DNA were removed from the solution. The remaining proteins were passed over the Z-DNA affinity column and then eluted with NaCl. Using both direct and competitive filter binding assays, we found that the eluted proteins bind to brominated poly(dG-dC) (Z-DNA) and poly(dG-m5dC) but not to poly(dG-dC) (B-DNA), native or denatured E. coli or calf thymus DNA, or brominated oligonucleotides. The proteins also bind to negatively supercoiled plasmids carrying Z-DNA sequences but not to relaxed or linearized plasmids in which the Z-DNA conformation is no longer present. Gel analysis reveals a mixture of several large proteins up to approximately 150,000 daltons. PMID- 6296849 TI - Isolated-patch recording from liposomes containing functionally reconstituted chloride channels from Torpedo electroplax. AB - Small unilamellar vesicles formed from purified phospholids by detergent/dialysis methods may be enlarged to 30-microns diameter by freezing and thawing. Very-high resistance seals were formed by applying a glass micropipette to the surface of these large liposomes, and single bilayer "patches" of membrane were isolated from the liposome surface while remaining sealed to the micropipette. The exogenous channel-forming peptides gramicidin and alamethicin induced characteristic single-channel fluctuation behavior in these excised patches held under voltage-clamp conditions. Large liposomes were formed from the small unilamellar vesicles made from cholate extracts of Torpedo electroplax plasma membrane vesicles. Isolated patches formed from these reconstituted membranes displayed current fluctuations due to single voltage-gated Cl- channels from non innervated-face membranes; the properties of these Cl- channels are identical to those observed in planar bilayer membranes after direct insertion from native membranes. This liposome-patch method combines the advantages of membrane protein incorporation into liposomes with high-resolution electrical recording methods and may provide a generally applicable approach to the study of integral membrane channel proteins after solubilization and reconstitution. PMID- 6296850 TI - Low-calorie diet prevents the development of mammary tumors in C3H mice and reduces circulating prolactin level, murine mammary tumor virus expression, and proliferation of mammary alveolar cells. AB - The effect of carlorie intake on the development of spontaneous mammary tumors in virgin C3H mice was studied. Only about 10% of the mice fed a low-calorie diet [10 kcal/day (1 kcal = 4.184 kJ)] since weaning developed mammary tumors, compared to about 60% of those mice that were reared on high-calorie diets (16 kcal/day or lab chow ad lib). In order to understand the mechanism by which a low calorie diet decreases the occurrence of mammary tumors in mice, we compared the sex cycle, the amounts of circulating thyroid-stimulating hormone (thyrotropin), growth hormone, and prolactin, the production of type A and B virus particles in the mammary glands, and the morphology of the mammary glands of mice fed low- and high-calorie diets. The amount of serum prolactin and the synthesis of type A and B particles in mammary tissues of mice fed a low-calorie diet was markedly decreased compared to those of age-matched mice fed high-calorie diets. In addition, in young mice fed a low-calorie diet, there were fewer mammary alveolar lesions than in mice fed a high-calorie diet, although the size of the lesions was similar. However, in older mice fed the high-calorie diet, the number and size of these lesions were greater than in the mice raised on the low-calorie diet. The other factors that we studied were not affected by calorie restriction. Our findings suggest that the reduction in serum prolactin level, mammary tumor virus production, and proliferation of mammary alveolar lesions associated with dietary calorie restriction is responsible for lowering the incidence of mammary tumors in mice. PMID- 6296851 TI - Normal serum and lipoprotein-deficient serum give different expressions of excitability, corresponding to different stages of differentiation, in chicken cardiac cells in culture. AB - Monolayers of cardiac cells from 11-day-old chicken hearts have different properties when maintained in fetal calf serum or in a lipoprotein-deficient serum (LPDS). Cells in fetal calf serum have a resting potential near -60 mV; the rate of rise of the action potential is low (less than 10 V/sec); the action potential and the contraction are essentially unaffected by tetrodotoxin (TTX); and the beating properties are unaffected by muscarinic agents. Cells in LPDS have a resting potential near -75 mV, and a fast rise of the action potential (approximately equal to 100 V/sec) that is drastically decreased by TTX with a parallel abolition of contraction, and the beat is blocked by very low concentrations of muscarinic agonists. Cells that are physiologically fully responsive to TTX and to muscarinic agents have receptors that remain stable 24 hr after protein synthesis is blocked, whereas cells that are physiologically unresponsive to TTX and muscarinic agents have receptors that are rapidly degraded with half-lives between 9 hr (TTX receptor) and 14 hr (muscarinic receptor). Differences in the physiological and biochemical properties are accompanied by changes in the cholesterol contents of the cell membranes. The properties of cardiac cells cultured in normal serum are similar to those found for cells of chicken hearts in the very early embryonic stage, whereas those of cardiac cells cultured in LPDS correspond to the late embryonic stage. PMID- 6296852 TI - Desensitization of steroidogenesis in cultured Leydig tumor cells: role of cholesterol. AB - We have reported previously that the human choriogonadotropin (hCG) receptors of cultured Leydig tumor cells can be down-regulated with hCG or mouse epidermal growth factor (mEGF) and that such down-regulation results in a loss of the steroidogenic response of the cells to hCG. Exposure of the cells to hCG, but not to mEGF, also resulted in a decrease in steroidogenic responses to cholera toxin and cAMP. The results presented herein show that the hCG-induced loss of steroidogenic response to cAMP is due to the depletion of intracellular cholesterol and that this depletion can be prevented by the addition of low density lipoprotein. Our results also show that after exposure of the cells to mEGF or hCG their steroidogenic response to hCG is limited by the number of hCG receptors, regardless of the presence of low density lipoprotein. PMID- 6296853 TI - Human alveolar macrophages produce leukotriene B4. AB - Human alveolar macrophages obtained by bronchoalveolar lavage were labeled overnight with [3H]arachidonic acid. The cells were stimulated with calcium ionophore A23187, and the 20:4 oxygenated metabolites released into the culture medium were identified by reverse-phase HPLC. Leukotriene B4 was the major 20:4 metabolite produced by these cultures. Leukotriene B4 was identified by its reverse-phase HPLC elution time, its UV spectrum, and its chemotactic and chemokinetic activities for neutrophils. In addition, the macrophage- and neutrophil-derived leukotriene B4 free acids and methyl esters were found to have identical HPLC retention times. PMID- 6296854 TI - Monoclonal antibody against angiotensin-converting enzyme: its use as a marker for murine, bovine, and human endothelial cells. AB - A monoclonal antibody has been prepared against rat angiotensin-converting enzyme (ACE). By selection for antibody binding to endothelial cells of bovine rather than rat origin we have obtained a reagent that has broad cross-species binding properties and that can at the same time serve as a useful marker for the surface of endothelial cells. The IgM-producing clone that we have established, alpha-ACE 3.1.1, has been grown in ascites form to yield ascites fluid that binds selectively to immobilized ACE at a greater than 1:10,000 dilution. By use of enzyme-linked immunosorbent assays, immunofluorescence histology, and flow cytometry, we have demonstrated the presence of ACE on endothelial cells of murine, bovine, and human origin. By means of a fluorescence-activated cell sorter (FACS-IV) we have been able to selectively isolate viable endothelial cells from a mixture of endothelial cells and fibroblasts. We believe the antibody will be useful not only for the selection and in vitro cultivation of endothelial cells but also as a tool for the identification and pharmacological study of ACE. PMID- 6296856 TI - cAMP-mediated decrease in K+ conductance evoked by serotonin and dopamine in the same neuron: a biochemical and physiological single-cell study. AB - The extracellular application of either serotonin or dopamine and the intracellular injection of cAMP all evoke in the same identified neurons of the snail Helix aspersa inward currents associated with a decrease in K+ conductance. The serotonin-, dopamine-, and cAMP-induced inward currents all show the same maximal amplitude. When the response to one transmitter is maximal, the response to the other is blocked. Using a single-cell microassay, we found that both serotonin and dopamine stimulate the adenylate cyclase [adenosine triphosphate pyrophosphate-lyase (cyclizing), EC 4.6.6.1] activity of the neurons giving the inward-current responses; on the other hand, the adenylate cyclase activity of a neuron that does not show the serotonin- and dopamine-induced currents was not stimulated by the transmitters. In contrast with the nonsummation of the maximal inward-current responses, the maximal stimulating effects of the transmitters on the enzyme activity are additive. The diterpene forskolin, which stimulates the adenylate cyclase activity of the single cells 9-fold, also evokes an inward current. We conclude that single snail neurons are endowed with independent serotonin and dopamine receptors linked to the adenylate cyclase. Activation of each of these receptors evokes a cAMP-mediated decrease in K+ conductance. The physiological interaction between the transmitters probably takes place at a late step in the chain of events leading from the increase in cAMP to the closing of the K+ channels. PMID- 6296855 TI - Radioimmunoassay for leukotriene B4. AB - A rabbit immunized with leukotriene B(4) [LTB(4); (5S,12R)-6, 14-cis-8, 10-trans icosatetraenoic acid] coupled to bovine serum albumin via the 12-oxy function of the lipid produced antibodies having an average association constant (K(a)) for [14,15-(3)H]LTB(4) of 3.2 x 10(9) M(-1) at 37 degrees C and in a concentration of 0.37 mug/ml of the immune plasma. When 10 mul of anti-LTB(4) and 3.9 nCi of [14,15-(3)H]LTB(4) (28 Ci/mmol; 1 Ci = 3.7 x 10(10) becquerels) were incubated in a volume of 250 mul, 50% inhibition of radioligand binding was achieved with 0.31 ng of LTB(4) and with 1.95 ng of (5S,12S)-6-trans-8-cis-LTB(4). The sulfidopeptide leukotrienes, LTC(4) and LTD(4), displaced the radioligand from this antibody with less than 1/100th the activity of LTB(4), and the diastereoisomers of 6-trans-LTB(4), 5-L-hydroxy-6-trans-8,11,14-cis icosatetraenoic acid (5-HETE), and three prostaglandins were minimally effective. The specificity of this radioimmunoassay was further shown by assessment of the immunoreactive products generated from calcium ionophore (A23187)-activated rat serosal mast cells and human neutrophils after reversed-phase HPLC. Resolution of the supernatants from each cell type yielded a single immunoreactive peak that coeluted with synthetic LTB(4) and quantitatively correlated with the physical measurement by integrated A(269) in that peak; UV-absorbing peaks eluting at other retention times were not immunoreactive. The immunoreactive LTB(4) generated averaged 4.6 ng per 10(6) rat mast cells and resolution of the supernatants by reversed-phase HPLC without a prior extraction step gave a recovery of 54%, validating the direct applicability of this sensitive and specific assay for LTB(4), a highly potent chemotactic factor, to unfractionated biologic fluids. PMID- 6296857 TI - Nucleotide sequence to the v-myc oncogene of avian retrovirus MC29. AB - Avian myelocytomatosis viruses are retroviruses whose oncogene (v-myc) induces an unusually wide variety of tumors, including carcinomas, endotheliomas, sarcomas, and myelocytomatoses. The viral gene v-myc arose by transduction of an undetermined portion of a cellular gene known as c-myc. In order to facilitate further studies of the functions of v-myc and c-myc and to permit detailed comparisons between the two genes, we have determined the nucleotide sequence of v-myc in the genome of the MC29 strain of myelocytomatosis virus. The v-myc domain in MC29 virus encodes a hydrophilic polypeptide with a molecular weight of 47,000, fused to a portion of the polyprotein encoded by the viral structural gene gag. The carboxyl-terminal half of the v-myc polypeptide is rich in basic amino acid residues. This feature may account for the DNA-binding properties of the hybrid gag-myc-encoded protein which would have a molecular weight of approximately 100,000, in accord with results from previous studies of the protein encoded by v-myc. The junctions between v-myc and the genome of the transducing virus are apparent but reveal no clues to the mechanism by which transduction might occur. PMID- 6296858 TI - "Onion skin" replication of integrated polyoma virus DNA and flanking sequences in polyoma-transformed rat cells: termination within a specific cellular DNA segment. AB - Replication of integrated polyoma virus DNA and flanking cellular sequences was studied in an inducible line of polyoma-transformed rat cells, designated the LPT line, that contains a single viral integration site. Chromosomal DNAs were purified from LPT cells treated with the virus-inducing agent mitomycin C and from untreated cells and were digested with restriction enzymes. The digests were analyzed by the Southern blotting technique. The virus DNA and a recombinant plasmid containing flanking cell DNA were used as hybridization probes. The analysis showed that mitomycin C treatment caused a more than 10-fold amplification of restriction fragments extending up to about 2.0 kilobase pairs into the cellular DNA flanking one end of the viral insertions, defined as the left joint. Fragments extending beyond this region were not amplified. These results showed that (i) integrated polyoma virus DNA undergoes multiple rounds of replication in mitomycin C-treated LPT cells and (ii) the replication extends into the flanking sequences and is arrested within a 0.40-kilobase-pair cellular DNA segment located about 2.0 kilobase pairs beyond the left joint. This segment may include a terminator of a normal cellular replicon. PMID- 6296859 TI - Abundant transcription of a cellular gene in T cells infected with human T-cell leukemia-lymphoma virus. AB - Human T-cell leukemia-lymphoma virus (HTLV) is a type C retrovirus associated with a subtype of mature T-cell malignancy in humans. HTLV also infects normal human cord blood mature T lymphocytes in vitro and induces a number of phenotypic changes in these cells, including their continuous growth and partial or complete independence of T-cell growth factor (TCGF). As part of our initial study designed to analyze gene(s) specifically activated by HTLV infection, we have isolated a recombinant DNA clone by differential screening of a cDNA library made from mRNA of a human T-cell lymphoma cell line producing HTLV. This cDNA identifies a single-copy gene in all human DNAs and a single mRNA species of 2.3 kilobases expressed at several hundred copies per cell in five HTLV-positive neoplastic T-cell lines. In addition, cord blood T lymphocytes infected with HTLV, but not the uninfected counterparts, express high levels of mRNA from this gene. A survey of different human hematopoietic cell types showed that this gene is expressed at low or undetectable levels (less than 10 copies) in human T, B, myeloid, or erythroid cell lines; in moderate amounts in lymphoid precursor (immature) cell lines; and in high amounts in lectin-activated mature T-cells, comparable to those of HTLV-infected T-cell lines. The precise function of this gene has not yet been determined. PMID- 6296860 TI - Demethylation and expression of murine mammary tumor proviruses in mouse thymoma cell lines. AB - Murine mammary tumor virus (MMTV) expression is analyzed in a T-lymphoid cell line (T1M1) sensitive to the killing effect of glucocorticoids and in two of its variants, one resistant (T1M1r) and one supersensitive (T1M1ss) to glucocorticoid induced lymphocytolysis. In the T1M1 line, MMTV is expressed and induced approximately 10-fold by short treatment with dexamethasone. Southern blot analyses of restriction enzyme digests of DNA from T1M1 cells reveal three proviruses similar to those of normal C57BL mouse tissue. In the T1M1ss line, which has retained functional glucocorticoid receptors, MMTV mRNA is inducible by glucocorticoids, while induction is reduced in the T1M1r line defective in glucocorticoid receptors. Moreover, the T1M1r line expresses a strikingly elevated basal level of MMTV mRNA in the absence of hormone. No rearrangements or superinfection have occurred in the variants, but all the regions containing 5' long terminal repeats are demethylated in the T1M1r variant although other sites of the provirus remain methylated. Because this variant was selected by prolonged treatment with dexamethasone, these observations raise the possibility that the continuous transcription of MMTV that occurred during this selection can result in glucocorticoid-induced demethylation of long-terminal-repeat sequences. PMID- 6296861 TI - Ascorbate-quinone interactions: electrochemical, free radical, and cytotoxic properties. AB - Standard midpoint potentials have been determined for p-benzoquinone, methoxy-p benzoquinone and 2,3-, 2,5-, and 2,6-dimethoxy-p-benzoquinones in aqueous solution. ESR studies have been made of the ascorbate and semiquinone radicals produced when these quinones interact with sodium ascorbate. Direct correlations are found between the electrochemical potentials, generated semiquinone lifetimes, and cytotoxic action in Ehrlich ascites-bearing mice. PMID- 6296862 TI - Modulator sequences mediate oxygen regulation of CYC1 and a neighboring gene in yeast. AB - Three transcripts from Saccharomyces cerevisiae--CYC1 mRNA (transcribed from the iso-I cytochrome c gene) and two RNAs of unknown function, designated tr-1 and tr 2-were identified by reverse Southern blot analysis and found to be regulated in response to oxygen. CYC1 mRNA and tr-1 accumulation occurred only in the presence of oxygen while tr-2 appeared only under anaerobic conditions. tr-2 was transcribed from a region approximately 1 kilobase 5' from the CYC1 coding sequence and in the opposite direction. tr-1 showed homology to the same region as tr-2 but was transcribed from elsewhere in the genome. Expression of tr-2 and CYC1 was observed to be normal in cells transformed with centromeric plasmids carrying the two genes. Mutant transforming plasmids were constructed in which a 400-base-pair region between tr-2 and CYC1 was either deleted or inverted. The deletion led to low-level nearly unregulated expression of both the CYC1 and tr-2 genes, suggesting that sequences upstream from both genes are important for their expression and regulation. The inversion mutation produced a reversed pattern of CYC1 regulation in which the mRNA was present in anaerobically grown cells but absent in the presence of oxygen, mimicking wild-type tr-2 regulation and suggesting that the CYC1 transcription unit is under the control of the translocated tr-2 modulator sequences. Models for the function of these modulators are discussed. PMID- 6296864 TI - Proton-hydroxide permeability of liposomes. AB - Liposome permeability to net proton-hydroxide flux was compared under two conditions that have produced extreme values in previous studies. In the first [Nichols, J.W. & Deamer, D.W. (1980) Proc. Natl. Acad. Sci. USA 77, 2038-2042] small pH gradients were permitted to decay near pH 7 and permeability coefficients in the range of 10(-4) cm/sec were obtained. In the second [Nozaki, Y. & Tanford, C. (1981) Proc. Natl. Acad. Sci. USA 78, 4324-4328] the decay rate of large pH gradients (approximately equal to 3 pH units) was measured and apparent permeability coefficients in the range of 10(-9) cm/sec were reported. We have repeated the conditions of the latter study and have found that the decay of large pH gradients produces a diffusion potential that limits net proton hydroxide flux. The permeability coefficients calculated from the flux are therefore underestimated and do not reflect intrinsic proton permeability. With the same liposome preparations under conditions in which small gradients decay so that diffusion potentials are negligible, the measured permeability coefficient is near 10(-4) cm/sec, as reported earlier. PMID- 6296863 TI - Anionic lipid headgroups as a proton-conducting pathway along the surface of membranes: a hypothesis. AB - Evidence has been gathering from several laboratories that protons in proton pumping membranes move along or within the bilayer rather than exchange with the bulk phase. These experiments are typically conducted on the natural membrane in vivo or in vitro or on fragments of natural membrane. Anionic lipids are present in all proton-pumping membranes. Model studies on the protonation state of the fatty acids of liposomes containing entrapped water show that the bilayers always contain mixtures of protonated and deprotonated carboxylates. Protonated fatty acids form stable acid-anion pairs with deprotonated fatty acids through unusually strong hydrogen bonds. Such acid-anion dimers have a single negative charge, which is shared by the four negative oxygens of both headgroups. The two pK values of the resulting dimer will be significantly different from the pK of the monomeric species, so that the dimer will be stable over a wide pH range. It is proposed that anionic lipid headgroups in biological membranes share protons as acid-anion dimers and that anionic lipids thus trap and conduct protons along the headgroup domain of bilayers that contain such anionic lipids. Protons pumped from the other side of the membrane may enter and move within the headgroup sheet because the protonation rate of negatively charged proton acceptors is 5 orders of magnitude faster than that of water. Protons trapped in the acidic headgroup sheet need not leave this region in order to be utilized by a responsive proton translocating pore (a transport protein using the proton gradient). Experiments suggest the proton concentration in the headgroup domain may vary widely and the anionic lipid headgroup sheet may therefore function as a proton buffer. Due to the Gouy-Chapman-Stern layer at polyanionic surfaces, anionic lipids will also sequester protons from the bulk solution at low and moderate ionic strengths. At high ionic strength metal cations may replace protons sequestered near the headgroups, but these cations cannot substitute for protons in the "proton conducting pathway," which is based on hydrogen bonding. PMID- 6296866 TI - Absolute quantum yields and proof of proton and nonproton transient release and uptake in photoexcited bacteriorhodopsin. AB - Using a sensitive differential ac conductance apparatus, we have measured transient ion movements in and the heating of bacteriorhodopsin suspensions after a light flash. The signal from the heating serves as an internal calibration of the absorbed photons and therefore the method gives the absolute quantum yield (phi) from a single measurement. At pH 4, H+ uptake precedes release, with phi = 0.4. By varying the buffer composition, we can prove that this signal is due to protons. At pH 8, however, the transient conductance increase is virtually independent of the buffer composition, showing that ions other than H+ are first released and then taken up by the purple membrane. If these ions are typical monovalent cations such as Na+ (lambda = 50 ohm-1 X cm2 X equiv-1), this process has a quantum yield of 2 or more at high salt concentrations. PMID- 6296865 TI - 31P NMR study of erythrocytes from a patient with hereditary pyrimidine-5' nucleotidase deficiency. AB - The composition of phosphate metabolites and the intracellular pH in erythrocytes from a patient with hereditary pyrimidine-5'-nucleotidase deficiency were examined using 31P NMR spectroscopy. Several resonances were identified in spectra from intact cells and from extracts. The 2,3-bisphosphoglycerate line intensities were normal but the NTP resonances were about twice normal due to the presence of millimolar quantities of pyrimidine phosphates. Several intense resonances were also observed in the diphosphodiester region of the spectrum. One compound contributing to these lines has been identified as cytidine diphosphocholine. The resonances of NTPs were in a position indicating that the additional triphosphates were also bound by Mg2+. Direct measurement shows that there is a nearly proportional increase in total cell Mg2+ in the patient's cells, in agreement with the interpretation of the spectra. The intracellular pH was about 0.2 unit lower in the patient's erythrocytes. This lower pH is due to the elevation in intracellular fixed negative charges and the shift in permeable anions consequent to the Donnan equilibrium. We suggest that the lower intracellular pH may explain the lower oxygen affinity of these cells in the presence of otherwise normal 2,3-bisphosphoglycerate levels and the increased Mg2+ triphosphates level, because the Mg2+ form of NTPs is known not to alter the oxygen affinity of hemoglobin under physiologic conditions. Furthermore, the lower intracellular pH can also explain the abnormalities in glycolytic intermediates observed for these cells. PMID- 6296867 TI - Modulation of adenovirus transformation by thyroid hormone. AB - We have examined the effect of triiodothyronine (T3) on de novo transformation of a cloned population of Fischer rat embryo fibroblasts (CREF) by a temperature sensitive mutant (H5ts125) of type 5 adenovirus and on the expression of the transformed phenotype in these cells. When CREF cells were grown in medium lacking T3 before, during, and after infection with H5ts125, the yield of transformed foci was half that in the cultures supplemented with 1 nM T3. Selective addition or removal of T3 during various phases of the transformation process indicated that the hormone exerted its maximal effect within 72 hr after viral infection. T3 was also required for optimal growth in agar of two clones of CREF cells previously transformed by type 5 adenovirus, wt-3A and ts-7E. The tumor promoter 12-O-tetradecanoylphorbol 13-acetate could substitute for T3 in enhancing growth in agar of wt-3A but not of ts-7E, suggesting that the promoter and T3 modify anchorage-independent growth by different mechanisms. Normal CREF cells and both of the transformed CREF clones grew equally well in monolayer culture in medium containing or lacking T3. Both of the transformed CREF clones contained a lower number of nuclear T3 receptors than did CREF cells and they bound somewhat lower levels of phorbol dibutyrate. These results indicate that thyroid hormone modulates an early stage involved in adenovirus transformation and that it also enhances the expression of the transformed state in previously transformed cells. PMID- 6296868 TI - 1 alpha,25-Dihydroxyvitamin D3 and 1 alpha-hydroxyvitamin D3 prolong survival time of mice inoculated with myeloid leukemia cells. AB - Syngeneic SL mice inoculated with murine myeloid leukemia cells (M1) all died of leukemia within 30 days. Treatment three times a week with 12.5-50 pmol per mouse of either 1 alpha,25-dihydroxyvitamin D3 [1 alpha,25(OH)2D3], the active form of vitamin D3, or its synthetic analog, 1 alpha-hydroxyvitamin D3 [1 alpha(OH)D3], considerably prolonged the survival time of mice inoculated with M1 cells. 1 alpha(OH)D3 was more effective than 1 alpha,25(OH)2D3 in increasing the survival time of the mice. 1 alpha(OH)D3 also increased the survival time of nude mice inoculated with M1 cells. The 1 alpha(OH)[3H]D3 administered intraperitoneally to tumor-bearing mice was converted very rapidly to 1 alpha,25(OH)2-[3H]D3. The chronic administration of 25 pmol of 1 alpha(OH)D3 to tumor-bearing mice for 30 days caused no appreciable hypercalcemia. These results indicate clearly that 1 alpha,25(OH)2D3 is effective not only in inducing differentiation of M1 cells in vitro, as previously reported [Abe, E., Miyaura, C., Sakagami, H., Takeda, M., Konno, K., Yamazaki, T., Yoshiki, S. & Suda, T. (1981) Proc. Natl. Acad. Sci. USA 78, 4990-4994], but also in prolonging the survival time of mice inoculated with M1 cells. PMID- 6296869 TI - Chromosomal assignment of the gene for the human beta 2-adrenergic receptor. AB - Chinese hamster fibroblasts (CHW) fail to physiologically respond to the beta adrenergic agonist isoproterenol with an increase in cellular cAMP. This unresponsiveness is due to a lack of beta-adrenergic receptors as indicated by an absence of specific binding of 125I-labeled hydroxybenzylpindolol (125I-HYP) to CHW plasma membranes. Preparation of somatic cell hybrids between CHW and human peripheral blood leukocytes led to the selection of a panel of 15 human-Chinese hamster cell hybrid clones, some of which had beta 2-adrenergic receptors (specifically bound 125I-HYP) and responded to isoproterenol (accumulated cAMP). Biochemical analysis of independent cloned cell hybrids indicated that beta 2 adrenergic receptor density and the intensity of the associated physiological response were closely correlated (r = 0.98). Both of these parameters were concordant in all cell hybrids with the presence of human chromosome 5. All other human chromosomes could be ruled out. These results suggest that the structural gene for the beta 2-adrenergic receptor is found on human chromosome 5. PMID- 6296870 TI - Association of the lethal yellow (Ay) coat color mutation with an ecotropic murine leukemia virus genome. AB - The dilute (d) coat color mutation on chromosome 9 is closely associated with an ecotropic murine leukemia virus (MuLV) genome [Jenkins, N.A., Copeland, N.G., Taylor, B.A. & Lee, B.K. (1981) Nature 293, 370-374]. DBA/2J mice homozygous for the reverse mutation to wild type at the dilute locus (d+2J) lack ecotropic virus specific sequences, suggesting that the dilute mutation was caused by virus integration. In the experiments described here, we analyzed the ecotropic MuLV DNA content of mice that collectively carry 10 different alleles at the agouti coat color locus (of chromosome 2) to determine whether any of these alleles also are associated with ecotropic virus sequences. Of the 10 alleles analyzed, one allele, lethal yellow (Ay), which is carried congeneically and heterozygously on C57BL/6J, 129/Sv, and LT/Sv mice, was closely associated with an ecotropic MuLV provirus. The close association of this provirus with the Ay allele suggests that this mutation also may be caused by virus integration. Furthermore, this association may be useful for molecular cloning and characterizing the many alleles at this locus. PMID- 6296871 TI - Several mechanisms can account for defective E alpha gene expression in different mouse haplotypes. AB - The murine Ia antigens, encoded by the I region of the major histocompatibility complex, are cell-surface glyco-proteins (consisting of alpha and beta polypeptides) thought to be involved in the control of immune responsiveness. Mice of haplotypes b, s, q, and f fail to express one of the Ia antigen complexes, the E complex, on the cell surface. We have attempted to determine at the molecular level how such a defect (or defects) might be generated. By using I region E alpha and A alpha gene probes for analyses of RNA and DNA structure, it was possible to conclude that at least three mechanisms can operate. Mice of haplotypes b and s bear a deletion in the E alpha gene, f haplotype mice synthesize predominantly an E alpha mRNA of aberrant size, and mice of the q haplotype seem to have a defect in RNA processing or a problem with mRNA stability, or both. PMID- 6296872 TI - Post-tetanic potentiation of acetylcholine release at the frog neuromuscular junction develops after stimulation in Ca2+-free solutions. AB - At many synapses, previous activity increases the amount of transmitter released by a single action potential. This potentiation of transmitter release is usually attributed to the local accumulation of the calcium ions that cross the axolemma during an action potential. We found that potentiated transmitter release can be observed at frog neuromuscular junctions after periods of repetitive stimulation in Ca2+-free solutions, if Ca2+ is restored after the tetanus. Potentiation is greater and more prolonged, the lower the level of extracellular K+. This component of potentiation may be due to Ca2+ that accumulates within the terminal in exchange for intracellular Na+. PMID- 6296873 TI - Phorbol diester receptor copurifies with protein kinase C. AB - The phorbol diester receptor present in the particulate fraction of rat brain was solubilized by divalent ion chelation in the absence of detergents. The soluble receptor was partially purified by (NH4)2SO4 precipitation, DEAE-cellulose, and gel filtration chromatography. The purified receptor required exogenous phospholipid for activity and displayed a Kd of 7 nM for [3H]phorbol 12, 13 dibutyrate. Biologically active phorbol analogs inhibited binding, whereas inactive analogs did not. The Ca2+-dependent, phospholipid-sensitive protein kinase C copurified with the phorbol receptor. Purified protein kinase C was activated directly by phorbol 12-myristate 13-acetate in the presence of phospholipid. PMID- 6296874 TI - Molecular cloning of a cDNA specific for the murine p53 cellular tumor antigen. AB - A cDNA library was constructed from sucrose-gradient-fractionated mRNA from SVT2, a mouse cell line transformed by simian virus 40. Polysomes containing the p53 messenger were specifically immunoprecipitated with monoclonal antibodies against the protein and used to prepare mRNA. This immunoselected mRNA, enriched 1,000-to 2,000-fold for p53-specific sequences, was used to make a cDNA probe and to screen the cDNA library. When approximately 10,000 colonies were screened by differential hybridization with probes made from immunoselected vs. nonenriched mRNA, a single clone was found that contained p53-specific sequences. The identity of this clone, termed pp53-208, was confirmed by the ability of its DNA to hybridize the mRNA coding for p53 (hybrid selection assay). When hybridized to a blot of EcoRI digested mouse DNA, the pp53-208 insert reacted with a single 3.3 kilobase band, suggesting that it is complementary to a single gene. PMID- 6296877 TI - Studies of inherited bleeding disorders to identify platelet membrane glycoproteins involved in adhesion and aggregation. PMID- 6296875 TI - Discriminatory inhibition of protein synthesis in cell-free systems by vaccinia virus transcripts. AB - The effect of vaccinia virus early transcripts on cellular (globin, HeLa, Chinese hamster ovary) and viral (vaccinia, encephalomyocarditis) mRNA function was studied in reticulocyte and wheat germ cell-free protein-synthesizing systems. Vaccinia virus transcripts of two size classes (8-10 S and 4-7 S), generated in vitro by viral cores, inhibited function of cellular and encephalomyocarditis virus mRNA but not that of vaccinia virus in reticulocyte lysate systems. Mild alkaline hydrolysis or micrococcal nuclease treatment of vaccinia virus in vitro transcripts resulted in a loss of their ability to inhibit protein synthesis directed by HeLa cell RNA. Vaccinia virus in vitro transcripts also selectively inhibited HeLa cell protein synthesis in wheat germ systems, suggesting that double-stranded RNA is not involved in this inhibition of protein synthesis. The addition, to the reticulocyte translating system, of cytoplasmic RNA obtained from infected cells in conjunction with cellular mRNA (globin, HeLa) resulted in the inhibition of synthesis of the globin or HeLa polypeptides with little or no effect on the translation of the vaccinia virus proteins. RNA extracted from vaccinia virions inhibited cellular but not vaccinia virus mRNA function when added to the reticulocyte lysate systems with uninfected or infected HeLa cell cytoplasmic RNA. PMID- 6296876 TI - Opioid peptide receptors and membrane biology. AB - The opioid peptides, beta-endorphin and the enkephalins, exhibit binding to brain tissue that is stereospecific, of high affinity, and saturable; from comparisons of the pharmacological potencies of a number of alkaloids with their abilities to displace peptide binding, it is concluded that peptide binding is probably relevant to pharmacological response for beta-endorphin, but not necessarily for enkephalins. The results of structure-activity studies indicate that both the (1 5) and a C-terminal sequence of the beta-endorphin molecule are necessary for both binding and pharmacological response, while pentapeptide conformation is related to enkephalin binding and pharmacological potency. The binding of opioid peptides possesses a brain regional distribution similar to that of alkaloids, with greatest enrichment in the striate, somewhat lesser amounts in the hypothalamus, thalamus, and amygdala, and least in the cortex and cerebellum; physiochemical properties, including inhibition by Na+, GTP, and by pretreatment of brain tissue with sulfhydryl reagents and proteolytic and lipolytic enzymes, are also similar to those of alkaloids. Other evidence, however, indicates that enkephalins and alkaloids bind to different sites: (1) differences in the abilities of enkephalins and alkaloids to displace labeled enkephalin and labeled alkaloid binding; (2) differences in relative pharmacological potencies of alkaloids and enkephalins in different systems; (3) small but significant differences in brain regional distribution and in certain physicochemical properties; and (4) selective protection of inhibition by irreversible reagents. This evidence, together with other data implicating both proteins and lipids in binding, has led us to propose a model of the beta-endorphin receptor in which the peptide binds to both an enkephalin site, located on a protein, and an alkaloid site, located on a lipid. Binding of enkephalins and beta-endorphin is related to a number of membrane-associated processes that are similarly affected by alkaloid binding, including changes in activity of adenylate cyclase and protein kinase, and in lipid metabolism and calcium ion disposition; some or all of these factors are presumably involved in the mediation of acute and chronic pharmacological effects. All of this work should be greatly aided by isolation of functionally active binding material, and recent work suggests that this breakthrough is finally possible. PMID- 6296878 TI - Somatic cell genetic analysis of human cell surface antigens. PMID- 6296879 TI - The adrenergic responsiveness of Down syndrome cells. AB - Platelets and cultured fibroblasts were used to examine the hormonal response of normal diploid and Down syndrome (trisomy 21) cells. An enhanced physiological response to catecholamine hormones was found in the trisomy 21 cells. This effect was specific for the adrenergic agonists since other hormones showed equivalent responses in the normal diploid and trisomy 21 cell types. Radioligand analysis of the alpha-adrenergic receptors in trisomy 21 platelets indicated no significant difference in receptor density or ligand affinity. Enzymatic analysis of the platelet adenylate cyclase activity in vitro showed no major changes in catecholamine sensitivity between the normal and Down syndrome preparations. Somatic cell genetic studies using human-hamster hybrids indicated that a gene coding for the beta-adrenergic receptor was not located on chromosome 21 but on human chromosome 5. These receptor, enzymic, and somatic cell genetic studies argue against a gene-dosage effect involving either the adrenergic hormone receptors or a component of the adenylate cyclase enzyme in the trisomy 21 cells. Identifying the gene(s) on chromosome 21 and the molecular mechanism(s) responsible for the exaggerated response of Down syndrome cells to catecholamines are the current objectives of this research program. PMID- 6296880 TI - Pseudohypoparathyroidism: deficiency of hormone receptor-adenylate cyclase coupling protein as a cause of hereditary hormone resistance. PMID- 6296881 TI - Actions of avermectin B1a on GABA nerves. AB - Avermectin B1a (AVM) is a macrolide derived from Streptomyces avermitilis. It possesses potent anthelmintic and insecticidal activities. It immobilizes nematodes by blocking the signal transmission from the central command interneurons to the peripheral motoneurons; this block can be reversed by the chloride ion channel blocker picrotoxin. The arthropods are paralyzed by AVM through drug inhibition of both inhibitory and excitatory postsynaptic potentials at the neuromuscular junctions. The mechanism of action is reduction of the muscle membrane resistance by opening the gamma-aminobutyric acid (GABA) controlled chloride ion channels in the membrane--an action also inhibitable by picrotoxin. AVM has specific and high-affinity binding sites in the mammalian brain synaptic membrane. The sites are concentrated in the cerebellum and closely associated with the GABA-benzodiazepine receptor-chloride ion channel complex. GABA release from brain synaptosomes is specifically stimulated by AVM. The density of available postsynaptic GABA receptors is increased by AVM--a process which requires chloride ion and is inhibited by picrotoxin. AVM also stimulates benzodiazepine binding and potentiates the in vivo diazepam activity of muscle relaxation. These findings add up to a conclusive demonstration of the mechanism of AVM action as a potentiation of the GABA action. They also point out the research on GABA nerve in invertebrates as an attractive approach to chemotherapy against nematodes and insects and suggest AVM as a very useful tool in isolating GABA receptors and in-depth understanding of GABA function. PMID- 6296883 TI - Assimilation and intracellular transport of lysosomal enzymes. PMID- 6296882 TI - Molecular interactions at the cell surface: role of glycoconjugates and membrane lipids in receptor recognition processes. AB - The present report has described studies using a particular receptor--the TSH receptor--to raise questions concerned with the role of glycolipids and glycoproteins in receptor recognition events and the relevance of lipid modulation of these components in regard to their functional expression. The importance of carbohydrates in other recognition systems will be even more eloquently detailed in subsequent presentations. It is clear, however, that we have gone beyond their recognition as important and are entered into a research stage where questions of how and why are center stage. It is equally clear, as evidenced by the monoclonal antibody data presented herein, that studies of these questions will provide new insight into the mechanism of disease states and will offer new and better diagnostic and therapeutic approaches. PMID- 6296884 TI - Prostaglandin regulation of murine mammary tumor virus production: a basis for some of the glucocorticoid and prolactin actions on mammary tumor cell cultures. AB - Hormonal regulation of mouse mammary tumor virus (MuMTV) production has been studied in cell cultures derived from mammary carcinomas of GR mice. The purpose of this study was to define the role of prolactin in dexamethasone-induced MuMTV production and to evaluate the mechanism(s) of action of these hormones. Results of our investigations in vitro establish a central role for prostaglandins, in particular PGF2 alpha. Prolactin treatment up to 1000 ng/ml increased the MuMTV production only slightly when added alone but a maximal stimulatory response was expressed when prolactin was added simultaneously with dexamethasone 2 micrograms/ml. This prolactin-related enhancement of MuMTV production was probably mediated by PGs since it was inhibited by indomethacin, an inhibitor of prostaglandin synthesis. When higher concentrations of indomethacin were used in dexamethasone-treated cultures, MuMTV production, although inhibited, was restored in part by addition of exogeneous PGF2 alpha. PGF2 alpha added alone stimulated the MuMTV production and, in combination with dexamethasone, showed a synergistic effect on stimulation of MuMTV production. Two compounds, PGF2 alpha and TXA2 (measured as TXB2), were produced in significant concentrations by these cultures. These findings suggest that PGF2 alpha exerts a regulating role on production of MuMTV in GR cells and indicate that the actions of both dexamethasone and prolactin are mediated at least in part by prostaglandins. PMID- 6296885 TI - Antithrombotic activity and the mechanism of action of trapidil (Rocornal). AB - Antithrombotic activity and the mechanism of action of trapidil were investigated, as compared with those of aspirin and dipyridamole. Trapidil at oral doses of 30 and 100 mg/kg inhibited arterial thrombosis in rats, while aspirin and dipyridamole at doses up to 300 mg/kg showed only a mild activity. This action may be explained by the fact that trapidil at concentrations ranging from 139 microM to 251 microM exerted 50% inhibition of platelet aggregation induced by ADP, arachidonic acid, thrombin or thromboxane A2 mixture, inhibition of platelet release reaction induced by ADP, arachidonic acid or thrombin, disaggregatory effect on aggregated rabbit platelets by arachidonic acid and potentiating action on antiaggregatory action of prostacyclin in vitro. In vitro actions of trapidil were apparently different from those of aspirin and dipyridamole. Trapidil also showed inhibition of platelet phosphodiesterase activity and thromboxane synthetase activity. Trapidil was expected to be an effective antithrombotic agent. The antithrombotic action of trapidil may be mediated by the inhibition of platelet function which is characterized by the inhibition of both thromboxane synthetase and phosphodiesterase activities, and by the potentiation of the antiaggregatory action of prostacyclin. PMID- 6296886 TI - Chronic administration of ethanol on pituitary and hypothalamic beta-endorphin in rats and golden hamsters. PMID- 6296887 TI - The need for a rapid and accurate viral diagnosis. PMID- 6296888 TI - Calcium ions, drug action and platelet function. PMID- 6296890 TI - Coronary physiology. AB - The major areas of normal coronary physiological research since Berne's 1964 review have been the measurement of ventricular transmural blood flow distribution with microspheres, the adenosine hypothesis of local metabolic control of coronary blood flow, and the autonomic control of coronary blood flow. There is an improved understanding of intramyocardial tissue pressure and extravascular compressive forces on coronary vessels. However, the unexpected finding of zero flow during a prolonged diastole with a coronary artery pressure of 40 mmHg (PZF) is a reminder that the physical forces, including vascular smooth muscle contraction, that determine coronary vascular resistance are incompletely understood. During normal circumstances, the left ventricular subendocardium probably receives more blood flow than the subepicardium does, but the difference is small. When the coronary circulation is compromised by stenosis or aortic valve lesions, the subendocardium is much more vulnerable to underperfusion than is the subepicardium. The coronary vasodilating effect of arterial hypoxia has been confirmed in many studies, but the role of tissue oxygen tension in local metabolic control of coronary blood flow during normoxia is unknown. The coronary vasodilating action of carbon dioxide has received renewed attention, but its role in local control is also unknown. The adenosine hypothesis has passed several critical tests, but despite much research the importance of adenosine in normal coronary regulation is not established. Local metabolic control of coronary blood flow probably involves more than just one factor, but a unified hypothesis has not been put forward. Sympathetic alpha receptor-mediated coronary vasoconstriction has been demonstrated by nerve stimulation and during a carotid sinus baroreceptor reflex. Sympathetic coronary vasoconstriction is capable of competing with local metabolic control to lower coronary venous oxygen tension under experimental circumstances, but its importance during normal resting conditions is not established. Parasympathetic muscarinic coronary vasodilation has been shown by vagal nerve stimulation, but a role for it during normal blood flow regulation has yet to be demonstrated. There have been elegant descriptive studies of the coronary blood flow response during excitement and exercise, where coronary blood flow increases pari passu with myocardial metabolism; however, there are also data that indicate a concomitant sympathetic vasoconstrictor effect during strenuous exercise. Overall there has been encouraging progress in coronary physiology. Inevitably new knowledge has focused old questions and presented new ones. PMID- 6296889 TI - Calcium ions, drug action and the red cell membrane. AB - Intracellular calcium regulates a number of membrane functions in the erythrocyte, including control of shape, membrane lipid composition and cation permeability. Measurement of total red cell calcium has yielded values between 5 and 15 nmol/ml cells, and these low values in part reflect the absence of Ca2+ containing organelles. Most intracellular Ca2+ is bound and the low cell ionized Ca2+ concentration (approximately 0.2 microM) is maintained by a combination of low membrane permeability and a powerful Ca2+ -pump. This pump has been identified with a (Ca2+ + Mg2+)-stimulated ATPase, and both Ca2+ transport and ATP splitting are stimulated by calmodulin, a low molecular weight protein which binds Ca2+ avidly and activates many Ca2+ -dependent enzymes. Both high and low affinity kinetics for Ca2+ pumping have been demonstrated, depending on the extent of binding of calmodulin to the pump. A stoichiometry of either 1 or 2 Ca2+ ions pumped per ATP molecule split has been shown, and the value may vary with the level of intracellular Ca2+. Phenothiazines, such as chlorpromazine inhibit the Ca2+ -pump by antagonizing the increment in activity produced by calmodulin. The passive inward leak of Ca2+ into erythrocytes can be quantitated by 45Ca2+ uptake into red cells whose Ca2+ -pump has been inhibited. Estimates of the Ca2+ permeability, based on unidirectional influx, yield values many orders of magnitude lower than for nucleated cells. Influx of Ca2+ into human erythrocytes occurs by a facilitated diffusion process, which can be inhibited by phenothiazines and the cinchona alkaloids. Calcium affects many membrane functions including cation permeability, lipid composition and some cytoskeletal interactions which may determine cell shape. Any rise in intracellular Ca2+ activates a specific K+ channel which normally makes little contribution to K+ fluxes. Kinetic studies of this process demonstrate either high or low affinity Ca2+ -activation of K+ efflux, with low affinity of the channel to Ca2+ being the probable state in vivo. Propranolol is the best known activator of Ca2+ stimulated K+ efflux, although the mechanism of stimulation is unclear. Like other tissues, red cells possess a Ca2+ -activated phosphoinositol phosphodiesterase. Although it has been suggested that the echinocytic shape change induced by Ca2+ is due to the hydrolysis of polyphosphoinositides, it seems more likely that this shape change results from an effect of Ca2+ on the macromolecular interactions of the cytoskeleton. Abnormal Ca2+ permeability may contribute to red cell destruction in a variety of diseases. For example, in sickle cell anemia a large Ca2+ influx occurs when cells are sickled under deoxy conditions, and moreover, the ability of the Ca2+ -pump to extrude the increment of cell Ca2+ is impaired. Thus, red cell Ca2+ is increased 3-7-fold above normal and this may contribute to the short survival of sickle red cells... PMID- 6296891 TI - Effects of some m-cholinomimetics and alpha- and beta-adrenomimetics on cyclic AMP content of the rat submaxillary gland. AB - The experiments were performed on four groups of rats of both sexes, anesthetized with pentobarbital: one control and three experimental groups. The experimental group was treated with carbachol 50 micrograms/100 g, isoprenaline 10 micrograms/100 g and phenylephrine 10 micrograms/100 g, respectively. The cyclic AMP was assayed by a R.I.A. method, at 4-6 min after administration of the drugs. The salivary secretion was estimated semiquantitatively by qualifying adjectives: very copious, copious and absent. In the control group the concentration of cAMP was 699 +/- 212 p.mol/g of wet tissue; carbachol administration resulted in an important and statistically significant increase (1030 +/- 205 p.mol/g; P less than 0.001) and isoprenaline increased the cAMP concentration more than twice (1414 +/- 368 p.mol/g, P less than 0.001). Phenylephrine decreased the level of cAMP to 481 +/- 79 p.mol/g, the decrease being statistically significant (0.02 less than p less than 0.05). PMID- 6296892 TI - Odor aversion learning by the rat fetus. AB - Rat fetuses were exposed to an odor stimulus on day 20 of gestation via amniotic injection and then injected with LiCl. In a CER paradigm, 10 day old pups were trained to approach an anesthetized dam in a runway for suckling reinforcement. When running speeds had stabilized the odor stimulus experienced in-utero was introduced into the test chamber. This odor took on aversive properties as a function of its pairing with LiCl, as evidenced by a decrease in running speed on CER trials and increases in the number of trials that were terminated because pups failed to traverse the runway. These data indicate that the fetal rat is capable of odor aversion learning. PMID- 6296893 TI - Further characterization of the anorexic action of orally-administered THIP, a GABA-analogue, in the rat. AB - Sprague-Dawley rats were used to further characterize the anorexic action of orally-administered THIP, a GABA-analogue. The anorexic action of THIP (5 or 10 mg/kg) was antagonized by prior subcutaneous injection of bicuculline (1 mg/kg), but not by prior subcutaneous injection of bicuculline-methobromide (1.5 mg/kg), strychnine-SO4 (0.75 mg/kg), pentylenetetrazol (25 mg/kg), or picrotoxin (1 mg/kg). Orally-administered GABA (50-300 mg/kg), bicuculline (1-10 mg/kg) or picrotoxin (1-10 mg/kg) generally did not inhibit food intake. These results indicate that the anorexic action of THIP is mediated by central GABA-receptors. PMID- 6296894 TI - Failure of ACTH to prolong extinction of a conditioned taste aversion in the absence of the testes. AB - Both corticotropin (ACTH) and testosterone prolong the extinction of a conditioned taste aversion in water-deprived intact male rats. An investigation was made to determine whether ACTH affects extinction in the absence of the testes and also to determine the effect of ACTH on serum testosterone levels. Water-deprived intact males showed prolonged extinction after ACTH injections; water-deprived gonadectomized males and intact females did not. All three of these groups showed elevated testosterone levels after ACTH administration, but testosterone levels were higher in the intact males than in the gonadectomized males or intact females. These results clearly show that in the absence of the testes ACTH is unable to prolong extinction. It is proposed that the increased level of testosterone following ACTH injection in water-deprived intact males is responsible for the prolonged extinction of a conditioned taste aversion. Although testosterone levels may increase in females and castrated males following ACTH injection, the increase is not sufficient to prolong extinction in these water-deprived animals. PMID- 6296895 TI - Syndactyly of the toes. AB - The experience gained through the management of 43 patients with syndactyly of the toes is presented. The incidence appears to be similar to that of syndactyly of the fingers. Type 1 syndactyly, or zygodactyly, always presented itself as a cosmetic problem; its correction is occasionally indicated and the procedure used is discussed. Type 2 syndactyly, or polysyndactyly, represents a functional problem and deserves surgical correction. My negative experience with the more complex procedures described for the correction of polysyndactyly is presented as well as my satisfaction with the simpler procedures. Suggestions for management are offered. PMID- 6296896 TI - The irritable bowel syndrome: soma and psyche. PMID- 6296897 TI - Who receives the aftercare? Utilization of services by discharged in-patients. AB - A prospective one-year follow-up of 216 in-patients showed that utilization of aftercare was significantly associated with certain clinical and social factors in the year after discharge from hospital. Rural domicile, a diagnosis of schizophrenia, long dependence on services, and ease of access to care comprised an aftercare dependency syndrome with substantial predictive power. Alcoholic and personality disordered patients were light users of services. Staff-patient relationships and attitudes of staff and patients to the need for aftercare were important. Depot neuroleptics were associated with high levels of aftercare for patients with chronic schizophrenia. The findings reveal the steady silting up of a new area service for the adult mentally ill by patients with a combination of social disadvantage and severe chronic psychosis. Family doctors played very little part in the delivery of aftercare. Some implications of the findings for the future development of community-based psychiatric services are discussed. PMID- 6296898 TI - Heroin and cocaine intravenous self-administration in rats: mediation by separate neural systems. AB - The hypothesis that separate neural systems mediate the reinforcing properties of opiate and psychomotor stimulant drugs was tested in rats trained to lever-press of IV injections of either cocaine or heroin during daily 3-h sessions. Pretreatment with the opiate receptor antagonist drug naltrexone produced dose dependent increases in heroin self-administration, but had no effect on the rate or pattern of cocaine self-administration. Similarly, pretreatment with low doses of the dopamine antagonist drug alpha-flupenthixol produced dose-dependent increases in cocaine but not heroin self-administration. High doses of alpha flupenthixol eliminated all responding for cocaine and slightly reduced heroin self-administration. The specificity with which the two antagonist drugs exerted their behavioral effects strongly suggests that independent neural substrates are responsible for the reinforcing actions of heroin and cocaine. PMID- 6296899 TI - Delayed effects of naloxone on responsiveness to environmental novelty in rats. AB - Two experiments were conducted to examine further the hypothesized involvement of endorphins in responsiveness to environmental novelty. In Experiment 1, rats were treated with naloxone hydrochloride (0.5--5.0 mg/kg, SC) before initial exposure to a novel arena (Day 1) and then retested in the area 24 h later (Day 2). Only naloxone (5 mg/kg) significantly affected Day 1 performance, producing a selective reduction in locomotor activity. However, compared to saline controls, all groups that had previously received naloxone showed marked reductions in both locomotor activity and rearing upon Day 2 retest. In Experiment 2, naloxone (0.5- 5.0 mg/kg) was without significant effect on performance in naive animals which had been injected on Day 1 but not exposed to the arena until Day 2. These data suggest that the delayed effects of naloxone relate specifically to the initial experience of environmental novelty rather than receptor changes or metabolite influences resulting from acute antagonist treatment. Results are discussed in relation to a possible action of naloxone upon mechanisms of attention and/or memory. PMID- 6296900 TI - Locomotor activity of rats after stimulation of the nucleus locus coeruleus region or after lesion of the dorsal noradrenergic bundle: effects of clonidine, prazosin and yohimbine. PMID- 6296901 TI - Antagonism of the anticonflict effects of chlordiazepoxide by beta-carboline carboxylic acid ethyl ester, Ro 15-1788 and ACTH(4--10). AB - The antagonism of the anticonflict effect of chlordiazepoxide (CDP) by beta carboline carboxylic acid ethyl ester (BCCE), Ro 15-1788 and ACTH(4--10) has been evaluated in the Geller-Seifter rat conflict test in which CDP increases punished (conflict), but not unpunished responding. BCCE (0.5--10 micrograms ICV) produced a dose-dependent reduction in the anticonflict activity of CDP. This was also significantly reduced by Ro 15-1788 (25 mg/kg IP) and a high dose of ACTH(4--10) (5 micrograms ICV). None of these test compounds had a marked direct effect on punished or unpunished responding in the dose used. These experiments provide further physiological support for the suggestion from binding studies that BCCE and Ro 15-1788 act on benzodiazepine receptors. However, the ability of ACTH(4- 10) to reduce the anticonflict effect of CDP may be by some other, possibly opioid, mechanism. PMID- 6296903 TI - Does conditioned nausea mediate drug-induced conditioned taste aversion? AB - Two antiemetic drugs were tested on the expression of taste aversions previously conditioned in rats with lithium, amphetamine or morphine. Neither prochlorperazine nor scopolamine administered prior to testing attenuated established aversions, although both drugs are known to have antiemetic effects in other species. Negative findings were obtained with a range of dose of prochlorperazine and scopolamine, with strong and weak aversions, with one- and two-stimulus tests, in a repeated one-stimulus extinction procedure, with between and within-group designs and with hooded, albino, male and female rats. The results do not support the widely accepted hypothesis that conditioned nausea mediates conditioned taste aversion. PMID- 6296902 TI - Role of dopaminergic neurotransmission in locomotor stimulation by dexamphetamine and ethanol. PMID- 6296904 TI - Lack of evidence for a role of endorphinergic mechanisms in mediating a discriminative stimulus produced by diazepam in rats. AB - Male Sprague-Dawley rats were trained in a two-lever food-reinforced procedure to discriminate between the effect of saline and diazepam (2.5 mg/kg). After acquisition of this discrimination, the ability of morphine to generalize, and naloxone to antagonize the diazepam discriminative stimulus was tested. The rats did not generalize the effect of morphine, and naloxone did not antagonize the diazepam discriminative stimulus whether it was given prior or subsequent to diazepam. These data suggest a lack of involvement of endorphins in mediating the discriminative stimulus property of diazepam. PMID- 6296905 TI - Using Ro 15-1788 to investigate the benzodiazepine receptor in vivo: studies on the anticonvulsant and sedative effect of melatonin and the convulsant effect of the benzodiazepine Ro 05-3663. AB - Both the anticonvulsant and sedative effects of diazepam (5 mg/kg) were reversed by subsequent administration of the suggested specific benzodiazepine antagonist Ro 15-1788. In contrast neither the seizure threshold raising or sedative effect of melatonin (200 mg/kg) was reversed by Ro 15-1788. Ro 15-1788 had no effect on the convulsant action of the benzodiazepine Ro 05-3663. These data therefore argue against the suggestion that melatonin produces its sedative and anticonvulsant effects in vivo by interacting with the benzodiazepine receptor, and also strengthens the suggestion that Ro 05-3663 does not act at this site. The use of Ro 15-1788 in demonstrating whether a drug acts in vivo at the benzodiazepine site to produce a pharmacological response is discussed. PMID- 6296906 TI - The effects of morphine and delta-9-tetrahydrocannabinol on motor activity in rats. AB - Acute treatment of rats either by high doses of morphine or delta 9 tetrahydrocannabinol (THC) decreased locomotor activity. Naloxone reversed morphine-induced depression completely and reversed THC-induced depression only partially. On day 3 of treatment, tolerance developed to the locomotor inhibitory action of THC or morphine and partial cross-tolerance was observed to the depressant action of THC. Naloxone slightly depressed locomotor activity in THC tolerant rats, but increased motor activity in morphine-tolerant rats. PMID- 6296907 TI - Evaluation of alpha-adrenergic responsiveness to clonidine challenge and noradrenergic metabolism in the affective disorders and their treatment. PMID- 6296908 TI - Cholinomimetic-induced co-release of prolactin and beta-endorphin in man. PMID- 6296909 TI - An electrophysiological analysis of the effects of cooling on autonomic neuromuscular transmission in the guinea-pig vas deferens. AB - The effects of cooling upon the excitatory junction potential (e.j.p.) and spontaneous e.j.p.s. in the guinea-pig vas deferens have been studied. A series of strict criteria have been used to select 'successful' penetration of smooth muscle cells by the micro-electrodes. Cooling the vas from 35 to 22 degrees C has little effect on the membrane potential. The e.j.p. is increased in size by cooling from 5 . 1 +/- 0.2 mV, n = 120 at 35 degrees C to 15 . 9 +/- 0 . 5 mV, n = 29 at 22 degrees C. Facilitation is less marked at lower temperatures. The effects of cooling are not due to any effects under the electrodes used to stimulate the innervation of the vas. The frequency and size of spontaneous e.j.p.s. is reduced by cooling. The significance of these observations is discussed. PMID- 6296910 TI - Determination of the radioprotective effects of topical applications of MEA, WR 2721, and N-acetylcysteine on murine skin. AB - Topical applications of MEA (beta-mercaptoethylamine or cysteamine), WR-2721 [S-2 (3-aminopropylamino)-ethylphosphorothioic acid], and N-acetylcysteine (NAC) were tested for their ability to protect the normal skin of the hind legs of mice against acute and late damage from single doses of 137Cs radiation. No significant protection was observed with either WR-2721 or NAC. MEA was shown to offer significant protection against acute skin damage in both buffered and unbuffered forms, but no significant protection against late contraction. The use of topical MEA on unanesthetized animals breathing carbogen (95% O2, 5% CO2) appears to give an enhanced level of radioprotection over that shown for anesthetized, air-breathing animals. PMID- 6296911 TI - Enhancement of heat sensitivity and modification of repair of potentially lethal heat damage in plateau-phase cultures of mammalian cells by diethyldithiocarbamate. AB - Diethyldithiocarbamate (DDC) is active both in vivo and in vitro in reducing the levels of enzymes such as superoxide dismutase (SOD) and glutathione peroxidase whose role in respiring cells is to remove toxic superoxide radicals and organic hydroperoxides. Although DDC, a copper-chelating agent, has been used to treat benign diseases, its potential as a heat sensitizer has not been fully explored. We have recently shown that the presence of 10(-3) M DDC for 2 hr causes a threefold reduction in the level of SOD in plateau-phase cultures of mammalian cells. At this concentration, the drug causes minimal toxicity but markedly affects both the shoulder and the slope of the heat survival curves. To explore another pathway of DDC sensitization, other than through reduced levels of SOD, we examined the repair of potentially lethal damage with and without DDC following exposure for 1 hr and 40 min at 43 degrees C. The repair, which progressed with a T 1/2 of about 10 hr, in either full medium or Hank's balanced salt solution (HBSS), in the absence of DDC, was completely blocked when DDC was added to the monolayers on completion of the heat exposure. DDC, in view of its ability to potentiate the effects of heat, is a potentially useful drug that could be used in an adjunctive setting with clinical hyperthermia. PMID- 6296913 TI - Attenuation of radiation- and drug-induced conditioned taste aversions following area postrema lesions in the rat. AB - The effects of lesions of the area postrema on the acquisition of radiation- and drug-induced (histamine and lithium chloride) conditioned taste aversions were investigated. The results indicated that area postrema lesions caused a significant attenuation of the aversion produced by pairing a novel sucrose solution with radiation (100 rad) or drug injection. Further, the area postrema lesions produced a similar level of attenuation of the taste aversion in all three treatment conditions. The results are discussed in terms of the implications of this finding for defining the mechanisms by which exposure to ionizing radiation can lead to the acquisition of a conditioned taste aversion. PMID- 6296912 TI - Skin radioprotection by 5-thio-D-glucose. AB - The radioprotective effect of 5-thio-D-glucose on mouse skin was studied. Intraperitoneal injection of A/J mice with 1.5 g/kg of 5-thio-D-glucose 2 hr prior to X irradiation of the foot reduced early foot skin damage through Day 40 postirradiation by a dose modification factor of 1.3 +/- 0.1. Similarly, late foot deformity during Days 60-90 postirradiation was reduced by a factor of 1.2 +/- 0.1. The radioprotective effect of 5-thio-D-glucose was also compared with that of WR-2721, an aminothiol radioprotector, in CDF1 mice. An intraperitoneal injection of 1.5 g/kg of 5-thio-D-glucose reduced early radiation-induced skin damage by a dose modification factor of 1.2 +/- 0.1 as compared to that of 1.5 +/ 0.2 by 0.65 g/kg of WR-2721 in this strain of mice. 5-Thio-D-glucose is also known to specifically kill and radiosensitize hypoxic tumor cells. Consequently, this drug may be a useful radiotherapy adjuvant, reducing normal tissue damage and enhancing tumor control by minimizing hypoxic protection. PMID- 6296914 TI - [Activity of Ca++-ATPase and enzymes of cycl AMP metabolism in the nervous tissue of rats at early stages of acute radiation sickness]. PMID- 6296915 TI - Reticular and raphe projections to the spinal cord of the North American opossum. Evidence for connectional heterogeneity. PMID- 6296916 TI - Brain stem projections to spinal motoneuronal cell groups in rat studied by means of electron microscopy autoradiography. PMID- 6296917 TI - Descending noradrenergic projections and their spinal terminations. PMID- 6296918 TI - Organization of descending serotonergic projections to the spinal cord. PMID- 6296919 TI - Descending pathways to the sympathetic preganglionic neurons. PMID- 6296920 TI - The organization of the direct cerebellospinal projections. PMID- 6296921 TI - Tectal control of spinal cord activity: neuroanatomical demonstration of pathways connecting the superior colliculus with the cervical spinal cord grey. PMID- 6296922 TI - Avian bulbospinal pathways: anterograde and retrograde studies of cells of origin, funicular trajectories and laminar terminations. PMID- 6296923 TI - Radioimmunoassay and tumor diagnosis. PMID- 6296924 TI - Humoral syndromes associated with cancer: ectopic hormone production. PMID- 6296925 TI - Comparative activity of natural mono-, di- and tri-hydroxy derivatives of arachidonic acid on guinea-pig lungs: myotropic effect and stimulation of cyclooxygenase activity. AB - The activity of natural 5,6-Dihydroxy-eicosatetraenoic acid (5,6-DiHETE; 2 isomers), 5S,15S-DiHETE, 8S,15S-DiHETE, 5S,12S-DiHETE, delta 6-trans-leukotriene B4, 12-epi-delta 6-trans-leukotriene B4, omega-hydroxy-leukotriene B4, omega carboxy-leukotriene B4, 15S-hydroxyeicosatetraenoic acid (15S-HETE), 12S-HETE, 5S HETE and 12S-hydroxy-heptadecatrienoic acid was compared to LTB4 on the guinea pig lung parenchymal strip and on the release of prostaglandins and thromboxanes by the perfused guinea-pig lungs. The omega-hydroxy-LTB4 appeared more potent than LTB4 both for inducing a contraction and for releasing prostanoids whereas the omega-carboxy-LTB4 was much less active on the parenchyma and did not release prostanoids at the dose used. All other hydroxy acids tested were either very weakly active or inactive in the two systems used with the exception of the 5,6 DiHETEs which showed significant activity. These di-hydroxy acids induced contractions of the lung parenchymal strip which could be blocked by FPL-55712 but were inactive on the guinea-pig ileum. The 5S-HETE, 12S-HETE and 15S-HETE were also tested for possible myotropic activity on selected smooth muscle preparations. Our results provide further informations on the structural requirements for LTB4 (and other hydroxy acids) actions on the guinea-pig lungs. PMID- 6296926 TI - [Effect of mouthwash of the "antidote" type on the condition of the oral cavity in workers of the aluminum plant]. PMID- 6296927 TI - [Ultrasound in the diagnosis of Wilms' tumor]. AB - Ultrasound (US) play a very important role in the diagnosis of Wilms' tumour. The authors analyze the findings in 12 children affected by this neoplasia. The resolution power of 4 different diagnostic levels is also analyzed. They are: a) mass identification; b) diagnosis of the tumour nature; c) diffusion balance; d) recurrence control. The most relevant resolution power lies in the diagnosis of nature and in the recurrence control. PMID- 6296928 TI - [Diagnostic reliability of scintigraphy in hepatic metastases of breast cancer]. AB - The authors have reviewed their experiences in determining the presence of liver metastases in 103 patients by radiocolloid scanning. The sensitivity of liver scanning proved to be quite low if the presence of focal defects of tracer's distribution was chosen as the diagnostic criterion. The inclusion of less restrictive criteria, such as liver enlargement or irregular distribution of the tracer, resulted in an higher sensitivity without lowering the predictive value of a negative scan. Using the more extensive diagnostic criterion, sensitivity, specificity and accuracy were in the range of 90%. Abnormal liver scans are common in patients classified as T3-T4 or N+ and their chances of to be "true positives" are high. Conversely, abnormal scans are seldom found in patients classified as T1-T2 or N0 and chances of "false positives" are high. PMID- 6296929 TI - [Suggestion for a methodology for studying uremic neuropathy]. PMID- 6296930 TI - Partial purification and properties of mevalonate kinase from neonatal chick liver. AB - Mevalonate kinase from neonatal chick liver has been partially purified by ammonium sulphate precipitation and Sephadex G100 and DEAE-cellulose fractionation. The kinetic characteristics agreed with the sequential mechanism suggested for the enzyme and provided apparent Km values of 0.01 mM for mevalonic acid and 0.25 mM for ATP. Partially purified mevalonate kinase from neonatal chick liver showed an absolute specificity for ATP. Mn2+ was a better activator than Mg2+ at low concentrations (0.1-1.0 mM). Higher Mn2+ concentrations produced a clear inhibition of mevalonate kinase. Likewise, addition of EDTA, with or without metal ions, clearly inhibited the enzymatic reaction. PMID- 6296931 TI - Ultracytochemical localization of Na+, K+-activated ATPase in the oocytes of Heterandria formosa Agassiz, 1853 (Pisces, Poeciliidae). AB - Using ultracytochemical methods, this paper studies the presence of Na+, K+ activated ATPase during oocyte development in Heterandria formosa. ATPase was discovered in all four oocyte stages, but its level was highest in stages II and III in which yolk formation took place. Na+, K+-activated ATPase was found on the membranes of both the yolk vesicles and the cortical granules and on the oocyte and follicular microvilli and the primary oocyte membrane, indicating that the microvilli were the sites of substance exchange between the follicle and the oocyte. PMID- 6296932 TI - Development of growth hormone receptors in rabbit and lamb liver after hypophysectomy. AB - The prolactin receptors of rat liver are pituitary-dependent, and previous studies have shown that prolactin itself plays a role in inducing and maintaining their presence. This study tried to determine if hepatic growth hormone (GH) receptors are comparably dependent on the pituitary. Young 47-day and older 116 day old rabbits were either hypophysectomized (H) or sham-operated (S). Hypophysectomy completely arrested the growth of the older rabbits but only reduced it by 50 p. 100 in the young ones. After 21 days, the specific binding (sb) of 125I-labelled human GH (hGH) to liver membranes was measured in H and S animals. The sb of hGH in H rabbits compared to S animals (older, young) was 14.9 p. 100 in older H rabbits and 45.5 p. 100 in young H animals. Similar studies in lambs showed that the sb of hGH in H lambs (compared to S animals) was 23.9 p. 100. When some H lambs were treated with 1 mg/kg of oGH or bGH 3 times per week, the sb of hGH was significantly increased to 56.1 p. 100 of the S levels. All changes in sb reflected changes in receptor number, as shown by the dose response binding curves. This study demonstrates that hepatic GH receptors in both the rabbit and the lamb are pituitary-dependent. The level in rabbits is correlated with the growth rate. Since GH receptor levels were partially restored in GH treated lambs, it is possible that GH plays a role in inducing its own receptors. PMID- 6296933 TI - [Growth hormone receptors in liver membranes of rats with renal insufficiency]. AB - Binding sites for human growth hormone (hGH) were studied in liver membranes of rats with chronic renal insufficiency (CRI) associated with a marked growth defect. A subtotal nephrectomy was performed in young female rats. One month after surgery, the rats with a plasma creatinine greater than or equal to 3 times that of controls were killed. The specific binding of 125I-hGH to microsomal membranes of rats with CRI was low. The number of binding sites rather than the binding affinity was affected; the number of both lactogenic and somatotropic sites was decreased. The sites of the plasma membranes as well as intracellular sites of Golgi fractions were reduced. No modification of the plasma GH and PRL was found. The hormone level did not appear to be the main regulating factor of the GH binding sites in this system. PMID- 6296935 TI - Adrenal maturation of the sheep fetus during late pregnancy. AB - The mechanism by which adrenal growth is achieved and steroidogenesis is stimulated in the fetal lamb during late pregnancy has been studied. Fetal plasma ACTH levels remained roughly constant between 14 and 3 days pre partum (298 +/- 38 pg/ml), then increased to a mean of 634 +/- 89 pg/ml. The rise in ACTH did not occur before the pre partum rise in cortisol. The increase in adrenal weight was achieved in three periods, one of DNA duplication between two phases of cellular hypertrophy. Cellular hypertrophy was accompanied by an important biosynthesis of membrane proteins. This development might partly explain the increased production rate of cortisol by the fetal adrenals during late pregnancy. At the same time, the number of ACTH receptors per 2 adrenals increased 5-fold. There was a close correlation between the levels of fetal plasma corticosteroids and the number of ACTH receptors during late pregnancy. It has been suggested that receptor modulation is one of the factors responsible for the enhanced sensitivity of the fetal adrenals to ACTH just before parturition. Preliminary results on the endocrine regulation of that development are also presented. PMID- 6296934 TI - [Short-term effect of prolactin on its receptor]. AB - It is known that prolactin (PRL) when injected chronically is able to increase the level of its own receptors. We have shown here that, after a single IV injection of PRL to lactating rabbits, total prolactin-receptor levels (measured after in vitro desaturation of injected prolactin by a MgCl2 treatment of the membranes) decreased from 41.6 to 18.4 p. 100 (specific binding) in 6 hrs. The same results were obtained in organ-culture of the rabbit mammary gland in the presence of prolactin; in addition, the down-regulation of prolactin receptors could be counteracted by lysosomotropic agents (chloroquine NH4Cl) in vitro. These results showed that prolactin could, at short term, induce a down regulation of its receptors, and that this effect might be related to an endocytosis of hormone-receptor complex and lysosome degradation. PMID- 6296936 TI - [Interaction of steroid hormones with their target cells]. PMID- 6296937 TI - [Murine teratocarcinoma: origin and relation to early embryonic cells]. PMID- 6296938 TI - Relations between the fertilizing ability, motility and metabolism of epididymal spermatozoa. AB - This paper compares the principal modifications taking place in spermatozoa during epididymal transit in several species. The relations between changes in the epididymal medium and modifications in the metabolism, motility and/or fertility of spermatozoa are reviewed with particular reference to spermatozoon forward motility. The fertilizing ability of motile testicular and epididymal spermatozoa has been described. It is suggested that the maturation process may induce a series of spermatozoal changes in cyclic AMP content, external membrane composition and nuclear structure throughout the genital tract; these changes would be under the control of the epididymal epithelium. The origins of these phenomena are practically unknown. PMID- 6296939 TI - [Role of vagal intestinal glucoreceptors in insulin regulation]. AB - This paper summarizes recent data on the existence of insulin nervous regulation by duodenal glucoreceptors which are connected to non-medullated fibres included in the vagal nerves. These glucoreceptors respond to intestinal perfusion of glucose or other carbohydrates, but other conventional mechanical stimuli are unable to activate them; therefore, they must be considered as true specific glucoreceptors. Their activation produces an immediate increase of insulin which precedes changes in glycemia. From bivagotomy experiments and from experiments which pharmacologically exclude the efferent parasympathetic or sympathetic fibres, it has been shown that this regulation is chiefly mediated by the vagal nerves. On the other hand, it was found that glucoreceptor afferents project on the hypothalamic nuclei (VMH and LHA) like the whole splanchnic and vagal afferents. The physiological importance and the role of insulin nervous regulation are discussed. PMID- 6296940 TI - [Interest in gnotoxenic systems for the study of host-microbial flora of the digestive tract]. AB - It is necessary to use experimental models in all studies of interactions between a host, its digestive tract microbial flora and the environment because these interactions are complex. The gnotoxenic animal, reared in an isolator as the axenic animal and harbouring a known microbial flora, constitutes either an analytic or mimetic experimental model. The gnotoxenic animal may be considered as an analytic model when used to determine which biotic or abiotic environmental factor of the host-animal plays a role in the intestinal ecosystem. The potential metabolic or immunologic role of a given bacterial strain in the intestinal ecosystem may be determined when the axenic animal is associated with this strain. The variation of the expression of the potential role of this strain in relation to the environment can be ascertained by diversifying the diet of the host or by introducing other bacterial strains into the intestinal ecosystem. The role of an association of strains in host physiology and host protection against potentially pathogenic target strains can also be studied using this analytic model. An analytic model is created by associating either isolated strains of the holoxenic or heated or diluted suspensions of holoxenic digestive tract flora. Axenic mice associated with these simplified flora are called meroxenic. The gnotoxenic animal is considered to be a mimetic model when it harbours a microbial flora isolated from an animal of a different species. The advantage of the mimetic model is that it provides an easy-to-use biological tool, i.e. gnotoxenic mice, to first determine the potential role of these microbial strains. The validity of the mimetic model is then tested by comparing gnotoxenic mice and gnotoxenic piglets or chickens. When all the gnotoxenic animals are given the same diet, this comparison permits an estimation of the animal-host role in the expression of the potential activities of microbial strains. The mimetic model, composed of gnotoxenic animals harbouring microbial strains of human origin, is the only experimental model which can be used to study the role of microorganisms in the intestinal ecosystem of man. PMID- 6296941 TI - [Comparison of goats and sheep with respect to the ingestion, digestibility and nutritive value of various diets]. AB - The voluntary food intake, organic matter, crude protein and crude fiber digestibility and nitrogen balance of 23 diets have been measured simultaneously using 5 to 7 wethers and 5 to 7 castrated billy-goats in digestibility trials. The goats ate more than the wethers (58 vs 46 g DM/kg W0.75). The digestibilities obtained for goats were highly correlated with those obtained for wethers, and the mean values were similar in both species: Organic matter: digestion coefficient (CUD)goats = 1.22 CUDwethers -14.5 r = 0.93 Crude protein: digestion coefficient (CUD)goats = 1.07 CUDwethers -4.8 r = 0.94 Crude fiber: digestion coefficient (CUD)goats = 0.56 CUDwethers +29.6 r = 0.78 The mean nitrogen balance for goats (226 mg/kg W0.75) was significantly higher than that of wethers (152 mg/kg W0.75). Based on the regression equations for each species, the calculated efficiency of proteins digestible in the intestine (PDI) was 55 p. 100, the endogenous urinary nitrogen 160 mg/kg W0.75 and the nitrogen maintenance requirement 1.8-1.9 g of PDI/kg W0.75. PMID- 6296942 TI - [Estimation of milk production in the nursing mare by labeling the body water of the foal]. AB - Female milk production was related to offspring water turnover, estimated by a tracer method (fig. 1). After deuterium oxide was injected into the blood of the offspring, we measured the decrease of its concentration in time. Milk intake was calculated by equations taking into account the weight gain of the offspring. The reliability of this method was checked in 28 bottle-fed lambs with known milk intake; the correlation between the actual intakes and our estimates was 0.98 with an error means of 5.6 p. 100. The error on the mean of 28 lambs was 0.5 p. 100 (table 1). The milk production of 6 heavy brood mares, estimated during the first week of lactation, varied from 12 to 20 kg per day between animals (table 2). This method did not interfere with the mother-offspring relationship nor with the usual suckling conditions; it can be used advantageously during the onset of lactation in species whose offspring suck frequently. PMID- 6296943 TI - Characterization and purification of a Ca2+ ion-activated neutral proteinase inhibitor in rabbit skeletal muscle. AB - This paper describes the isolation, purification and properties of a specific inhibitor of calcium-activated neutral proteinase (CaANP) in rabbit skeletal muscle. The inhibitor was a thermo-acid-stable protein degraded by trypsin and chymotrypsin and seemed to contain two polypeptide chains with molecular weights of 70 000 and 13 000 daltons. Maximal inhibitory activity was obtained at neutral pH. High salt concentrations were needed to suppressinhibition. Inhibitor concentration had no effect on the optimal Ca++ ion levels for CaANP. These experiments also show that enzyme inhibitor association was instantaneous and did not need any incubation. PMID- 6296944 TI - [Dietary components and digestive motility]. PMID- 6296945 TI - Participation of glucocorticoids in the regulation of plasma LH levels in the male domestic duck (Anas platyrhynchos L.). AB - Adult male domestic ducks were treated in autumn for 14 consecutive days with either (i) long-acting ACTH (20 IU/day), (ii) corticosterone (25 mg/day), (iii) dexamethasone (0.25 mg/day) or (iv) a control solution. Blood samples were obtained from all birds after 1, 3, 7 and 14 injections of hormones; on each of these occasions, a sample was obtained 3 times during the day. The 3 hormonal treatments induced a marked and transient (observed on the 3rd and 7th, but not on the 1st and 14th days of injection) elevation of the LH plasma levels. This increase was not associated with any alteration of the testosterone plasma levels. The results are discussed in view of our knowledge of the influence of adrenal hormones on the pituitary-gonadal axis. PMID- 6296946 TI - Influence of dietary essential fatty acid level on fatty acid composition in peripheral nerve and muscle. AB - Two groups of rats of different lines were divided into two lots. The four lots, whose dams received a fat-controlled diet, were fed with diets containing different levels of essential unsaturated fatty acids, namely linoleic (18:2 (n 6)) and linolenic (18:2 (n-3)) acid, in the oil. Nerve and muscle fatty acids were analyzed at adulthood. When the linolenic acid level was less than 0.6 p. 100 of the total fatty acids in the diets (peanut oil and sunflower oil), the level of long-chain polyunsaturated fatty acids (PUFA) of the (n-3) series decreased. This decrease was associated with an increase in the PUFA of the (n-6) series. Total PUFA ((n-3) + (n-6)) remained similar in the same group of rats, whatever the diet. On the contrary, when dietary linolenic acid reached 7.6 p. 100 (soya oil) or 8.5 p. 100 (rapeseed oil) of the total fatty acids, in the presence of a very high linoleic acid level, the PUFA of the (n-6) series decreased and that of the (n-3) series increased in nerve and especially in muscle. It appears that preferential PUFA biosynthesis from linolenic acid occurred when the ratio (n-6)/(n-3) was low enough in the dietary oil. PMID- 6296947 TI - Dietary fiber and cholesterol and bile acid metabolisms in axenic (germfree) and holoxenic (conventional) rats. I. -- Effect of wheat bran. AB - Axenic (GF) and holoxenic (CV) rats were given a semi-synthetic diet containing no fiber (0) or 10 p. 100 bran (B). The GFO, GFB, CVO and CVB groups were compared to assess the action of the flora and the bran. The bran diet did not change body growth, food intake or cecal water content. It caused only a slight decrease in the digestive utilization of the diet in the holoxenic rats and a decline in cecal weight in the axenic rats. The bran diet modified unabsorbed cholesterol transit in both types of rats, and slightly altered dietary cholesterol absorption which was a little lower in axenics than in holoxenics. In the former (GF), the bran-containing diet dit not change either the plasma or the hepatic cholesterol concentration. In the latter (CV), it increased plasma cholesterol which was lower in CVO rats than in the other three groups and decreased hepatic cholesterol which was thus lower in the CVB lot than in the others. The bran diet reduced fecal cholesterol elimination in axenic and holoxenic rats. This decrease was a little higher in the latter. The digestive tract microbial flora was thus implicated in the effect of bran on those characteristics of cholesterol metabolism. The bran did not change fecal bile acid composition in the axenics. In the holoxenics, it contributed to make this composition uniform because without bran, individual compositions varied. The microbial flora thus changed many aspects of bran action on cholesterol and bile acid metabolisms. But total cholesterol and bile acid elimination was only slightly affected by those two factors. PMID- 6296948 TI - Distribution of oxidases in the testis of buffalo, goat and ram: an histochemical study. AB - Peroxidase, monoamine oxidase (MAO) and cytochrome oxidase (CCO) have been histochemically localized in the testis of buffalo, goat and ram. The results in these three species were more or less similar. Peroxidase was localized only in the interstitial tissue and could be used as a marker enzyme for this testicular compartment. MAO and CCO were present in both the interstitial tissue and the seminiferous tubules. The detailed patterns of MAO and CCO distribution were also similar and showed cyclic changes in the spermatids which have not been reported before. The results have been compared and contrasted with those of previous workers and their possible physiological significance discussed. PMID- 6296949 TI - Dietary fiber and cholesterol and bile acid metabolisms in axenic (germfree) and holoxenic (conventional) rats. II. Effect of pectin. AB - Axenic and holoxenic rats were fed with a semi-synthetic diet containing 5 p. 100 pectin as the only fiber. Although the microbial flora of the digestive tract converted most of the bile acids in holoxenic rats, it hardly changed the other characteristics of cholesterol and bile acid metabolisms, except intestinal bile salt pools which were much larger in the axenic rats. When the pectin-containing and pectin-free diets were compared, it appeared that the former diet increased cecal weight, especially in axenic rats, but slightly modified cholesterol and bile acid metabolisms. However, the values of some characteristics of those metabolisms varied individually in the holoxenic rat fed with the pectin containing diet, while they were very homogeneous in the other groups. This heterogeneity might result from pectin-digestive tract microbial flora interactions which would vary among individuals. PMID- 6296950 TI - [Effect of diets low in phosphorus and high in dietary calcium on phosphatemia and phosphorus and calcium levels in rumen contents of sheep]. AB - Four semi-purified diets with different calcium (Ca) and phosphorus (P) contents were given to adult sheep at a level of 1 kg daily plus 100 g of straw. The phosphorus was supplied as dicalcium phosphate, almost insoluble in rumen fluid. In the preliminary period, all the animals (16) were fed the test diet (NCa-NP) containing 8.4 g of Ca and 3.3. g of P. In the experimental period (table 3) two trials were carried out. In trial 1, two groups of 5 sheep each were given for 5 weeks a diet low in P (1 g/kg) and either low (2 g/kg) (diet BCa-BP) or high (10 g/kg) (diet HtCa-BP) in calcium. In trial 2, which lasted for 19 weeks, the HtCa BP diet was compared to a high Ca (10 g/kg) normal P (3.3 g/kg) diet (diet HtCa NP); two groups of 2 sheep each were fed both these diets every 3 hours, and one group (HtCa-BP) was fed twice daily. The concentration of ultrafilterable inorganic Ca (CaiU) in the rumen contents was related to the Ca level of the diet (tables 4, 5). However, individual variations were high and CaiU concentrations were negatively correlated to rumen pH (fig. 7). When diets adequate in P were fed, ruminal concentrations of ultrafilterable P (PiU) were rather high (400-490 mg/l), showing the importance of endogenous P supply in the rumen. With an adequate P supply, a high Ca intake had no effect on either plasma or ruminal PiU. When low P diets were fed, there was a steady decrease in plasma Pi from 6 to 3.5 mg P/100 ml and in ruminal PiU from 500 mg/l to values lower than 300 mg/l up to 5-6 experimental weeks. These decreases were greatest with high Ca intake (HtCa-BP diet) (figs. 2, 3), some animals being markedly affected (ruminal PiU values lower than 20 mg/l). However, the decreases in ruminal PiU were less pronounced with frequent feeding than with feeding twice a day. Considering all the results obtained, ruminal PiU concentrations were positively correlated with plasma Pi concentrations (r = + 0.77; n = 75) (fig. 8). Therefore, high Ca intake can enhance the response to P deficiency and reduce further plasma Pi concentrations and levels of available P in the rumen. Microbial P requirements may not be satisfied in such conditions, which would reduce ruminal microbial digestion in some animals. PMID- 6296951 TI - Application of recombinant DNA techniques to neurogenetic disorders. PMID- 6296952 TI - Feline viral rhinotracheitis virus: explant and cocultivation studies on tissues collected from persistently infected cats. AB - Tissues from 16 feline viral rhinotracheitis (FVR) carrier cats and 15 controls were examined by the techniques of homogenisation, cocultivation and explant cultivation. The tissues examined included trigeminal ganglion and maxillary nerve, olfactory lobe and nerve endings, tonsils, submandibular lymph node, spleen and parotid salivary gland. Most of the cats were shedding FVR virus in the oropharynx at the time the tissues were collected. No evidence of a persistent FVR virus infection was found in any of the tissues. Trigeminal ganglion explant cultures from six FVR virus carrier cats were shown to be latently infected with feline syncytia forming virus by treatment with 5 iododeoxyuridine. Five control cats had trigeminal ganglion explant cultures similarly treated but produced no evidence of infection. The possible significance of this is discussed. PMID- 6296953 TI - Importance of competitive binding in the detection of antigen specific bovine isotypes and subisotypes by the micro ELISA. AB - An enzyme-linked immunosorbent assay was developed to detect and quantify the specific bovine immunoglobulin class response to Trypanosoma theileri, Dictyocaulus viviparus and infectious bovine rhinotracheitis virus. Comparative measurement of the specific immunoglobulin classes in whole serum was achieved using monospecific rabbit antibovine IgG1, IgG2 and IgM, followed by a goat anti rabbit Ig-enzyme conjugate (antiglobulin ELISA). The results obtained in the antiglobulin ELISA compared favourably with the standard (or indirect) ELISA using the purified immunoglobulins. Competitive inhibition between specific immunoglobulins of different isotypes and subisotypes was the major disadvantage of the antiglobulin ELISA. This latter assay failed to detect specific IgG2 against T theileri antigen in both calf and adult whole serum. The inability to detect the specific IgG2 was a result of competitive inhibition by specific IgG1. However, competitive inhibition between specific immunoglobulins was not observed in either of the other test systems using D viviparus and infectious bovine rhinotracheitis virus antigens. PMID- 6296954 TI - Equine infectious anaemia: detection of antibodies using an immunofluorescence test. AB - An indirect immunofluorescence test for detecting antibodies to equine infectious anaemia virus is presented. Using monolayers of equine dermal cells within a defined period after infection, discrete fluorescent spots were observed in the cytoplasm of as many as 95 per cent of the cells. These inclusions appeared as ring-like structures when high titred sera were employed but became spots when the sera were diluted. Cells showing optimal antigen fluorescence were used immediately or after storage at -70 degrees C. The fluorescence test detected lower levels of antibody than the immunodiffusion test, and results were available in less than two hours. It should be useful as a confirmatory assay with the immunodiffusion test. PMID- 6296955 TI - Air sampling of pigs infected with foot-and-mouth disease virus: comparison of Litton and cyclone samplers. AB - The air in looseboxes containing groups of pigs in the acute stage of foot-and mouth disease was sampled simultaneously with two air-sampling devices: a large volume sampler (Litton) and a cyclone sampler. Although the cyclone sampler was slightly less efficient at trapping airborne virus it was easier to operate. When pigs were sampled individually within a 610 litre cabinet using the cyclone sampler, the mean recovery of virus over a 24 hour period was log10 8 X 6 TCID50 per animal. This figure is four times higher than the maximum amount estimated in previous studies in which groups of pigs held in looseboxes were sampled with a Litton apparatus. PMID- 6296956 TI - [Total ethmoidectomy by a mixed approach: extradural frontal and paralateral nasal]. PMID- 6296957 TI - [Growth hormone. II. Clinical application]. PMID- 6296958 TI - [Early, intermediate and advanced stomach cancer: morphological features]. PMID- 6296959 TI - [Preleukemic states]. PMID- 6296960 TI - [Subacute polioencephalomyelitis with cancer. 6 anatomo-clinical cases and review of the literature]. PMID- 6296961 TI - Nonbacterial pneumonia after allogeneic marrow transplantation: a review of ten years' experience. AB - Pneumonia due to causes other than bacterial or fungal infection has been a frequent complication of allogeneic marrow transplantation. Data on 525 patients who received allogeneic marrow transplants during a 10-year period were reviewed. Of these patients, 41% developed pneumonia; this incidence was significantly higher than that among recipients of syngeneic (twin) transplants. Cytomegaloviral pneumonia (85 cases) and idiopathic pneumonia (63 cases) occurred most commonly. The incidence of pneumonia was higher among older patients, among patients who received transplants because of hematologic malignancy, and among patients with aplastic anemia who received total-body irradiation or procarbazine plus antithymocyte globulin for conditioning before transplantation. The development of cytomegaloviral pneumonia was unrelated to the serologic characteristics of either the patient or the donor before transplantation, and an increase in the titer of antibody to cytomegalovirus did not significantly improve the chances for survival. Mortality from all forms of pneumonia was high. Until effective means for prevention or treatment of cytomegaloviral and idiopathic pneumonia become available, the occurrence of these infections will continue to limit the success of allogeneic marrow transplantation. PMID- 6296962 TI - Immunology of Entamoeba histolytica in human and animal hosts. AB - Although Entamoeba histolytica induces humoral and cellular immune responses in both human and animal hosts, there is no indication of postinfection immunity in humans; in contrast, several other mammals are protected by prior infection or immunization. The exacerbation of the disease by immunosuppression suggests a protective function of still-unknown defense mechanisms. Specific local and circulating antibodies are produced regularly during invasive amebiasis. Although serum antibodies, together with complement, are lytic to the trophozoites in vitro, the poor correlation of these antibodies with resistance contradicts a protective capacity in vivo. The parasite may evade harm by shedding antigen antibody complexes from its surface. Demonstration of immediate-type skin reactions, elevated IgE titers, and specific antiamebic IgE suggests that anaphylaxis occurs. The function of the anaphylactic reaction in pathology and resistance remains to be studied. Delayed hypersensitivity parallels healing or resistance and is retarded in human hepatic amebiasis. This observation is consistent with a protective role of cell-mediated immunity. PMID- 6296963 TI - A review of the parasite cellular mechanisms involved in the pathogenesis of amebiasis. AB - Amebiasis, a significant cause of morbidity and mortality on a global scale, is caused by invasion of the colonic mucosa by the cytolytic protozoan parasite Entamoeba histolytica. A review of the investigations into the mechanisms by which the parasite causes invasive disease is presented, including the relevant history, epidemiology, and pathology of amebiasis, the biology of the parasite, in vivo and in vitro studies of the pathogenesis of disease, the host immune responses, and possibilities for prevention and biologic control. New insights into the cell biology of E. histolytica, especially its adherence and cytolytic properties, offer several new approaches to biologic control of this invasive, destructive parasitic infection. PMID- 6296964 TI - Amebiasis: host-pathogen biology. AB - Invasive amebiasis caused by Entamoeba histolytica, and particularly amebic liver abscess, is a major public health problem in Mexico and some other countries because of the high incidence and mortality of this disease. This paper first discusses the pathogenic effect of E. histolytica and the defensive response of the host and then reports studies concerning the experimental induction of protective immunity to amebic infection. The pathogenic effect of E. histolytica is probably initiated by a lectin-mediated adhesion of trophozoites to target cells; the adhesion is followed by cytopathic activity and phagocytosis by the ameba. The defensive response is characterized by humoral and cellular immune reactions. Humoral immunity manifests itself by specific circulating antibodies useful in the diagnosis and seroepidemiology of amebiasis. Cellular immunity is shown by several characteristic reactions. Experimental induction of immunoprophylaxis with E. histolytica antigens represents the first stage in the development of a vaccine against E. histolytica for use in humans. PMID- 6296965 TI - Differences among moxalactam, cefoperazone, and cefotaxime in activity against multiple-antibiotic-resistant gram-negative bacteria. AB - In vitro evaluations have shown that moxalactam, cefoperazone, and cefotaxime, three new beta-lactam antibiotics, have certain potency and antibacterial spectral characteristics in common. Results of parallel tests of these antibiotics against routine clinical isolates have suggested that these compounds could be tested with one disk that is representative of this spectral class. The key requirement for this class concept in disk testing is that there be no bacterial resistance mechanism that can operate against one, but not all, members of the proposed class. The results of an 11-center study of approximately 500 isolates of multiple-antibiotic-resistant Enterobacteriaceae responsible for nosocomial infections illustrated that cross-resistance to moxalactam, cefoperazone, and cefotaxime is highly variable from center to center. Thus, the spectral class concept of disk susceptibility testing would be inappropriate for this group of drugs. PMID- 6296966 TI - Comparative multiple-dose pharmacokinetics of cefotaxime, moxalactam, and ceftazidime. PMID- 6296967 TI - Comparative pharmacokinetics of moxalactam, cefoperazone, and cefotaxime in normal volunteers. AB - The pharmacokinetic parameters of moxalactam were compared with those of cefoperazone and cefotaxime in normal volunteers in a crossover manner. Following 30-min intravenous infusions of 2 g of each of the three antibiotics, serum levels at 1 hr were slightly lower for moxalactam (88 micrograms/ml) than for cefoperazone (112 micrograms/ml) but more than three times those obtained for cefotaxime (29 micrograms/ml). By 8 hr, levels of moxalactam (9.2 micrograms/ml) were slightly higher than those of cefoperazone (6.5 micrograms/ml), and levels of cefotaxime in serum were unmeasurable (less than 1 micrograms/ml). These values reflect differences in half lives of the three agents. Peak serum levels following intramuscular injection and serum levels during constant intravenous infusion were similar for moxalactam and cefoperazone because of counterbalancing differences in the apparent volume of distribution and rates of elimination of the two antibiotics. Serum levels of cefotaxime were much lower than those of the other two antibiotics primarily because of the rapid elimination of cefotaxime from the body. The kidney was the major route of excretion of moxalactam, whereas extrarenal mechanisms were more important for elimination of cefoperazone. These differences in pharmacokinetics may have significant implications for the clinical use of these new antibiotics. PMID- 6296968 TI - [Orientation in the treatment of paracoccidioidomycosis (South American blastomycosis)]. PMID- 6296969 TI - [Structure and functions of pulmonary endothelia]. PMID- 6296970 TI - [Current polychemotherapy of bronchopulmonary cancers]. PMID- 6296971 TI - [Evolution of chronic pulmonary tuberculosis and the possibility of preventing chronicization (in the Iasi judicial district)]. AB - The instantaneous prevalence of patients with pulmonary tuberculosis has been evaluated; the patients were included in the dispensarization group I between 1974 and 1980. Initially there was a decrease of the prevalence of the patients in Group I B but over the last years this was stable at 11.7-13.5 0/0000 inhabitants. The decrease which had been obtained at first was due to the immediate effect of repeated treatments with new tuberculostatic drugs resulting in sterilization and inclusion in the Group II of dispensarization of many of the preexisting patients while the ceiling which occurred later was determined by the quantitative balance of therapeutic failures and relapses entering the Group I B and cases which left the Group as a result of medical solutions, death or change of address. It was also noted that most of the patients in whom presence of the bacilli had been determined by microscopic techniques and by cultures constituted a relatively stable group over the entire period of the study, and oscillated between 11 and 17.5%, and 56.5 and 65.9% respectively. The quantitative balance established over the last years between the "entrances" and the "exits" from Group I B may be altered firstly by lowering the number of failures and relapses after the first treatment. The nature of unfavourable factors which influence the prognosis may be classified as follows: bio-psycho-social implications (males- 81.5%, age--between 30 and 50 years, 51.4%, lack of occupation, 31.5%, low educational level, 80.1% chronic ethyl intoxication 51.4%, psychoses, 12.7%, other associated diseases, 34.3%, and primary drug resistance, 7.1%); consequences of a late detection; deficiencies in the rhythm and the duration of the first treatment with tuberculostatic drugs (non-cooperation--64.2%, irregularities--38.5%, interruptions--42.8%, secondary drug resistance--14.2%). PMID- 6296972 TI - [Effectiveness of the examination of the different categories of contacts of pulmonary tuberculosis patients as a part of epidemiological investigations]. AB - In view of evaluating the global efficiency of the epidemiologic investigation in relation to its area an analysis was carried out of 127 investigations involving 1 252 tb contacts. The contacts were classified in three groups: intrafamilial and intradomiciliary, intrafamilial but outside the home, and extrafamilial and outside the home; the results of the examinations were compared between them on the one hand and with the proportion of allergic children (51.6%) and the instantaneous prevalence of bacilli eliminating cases (0.31%) on the other hand. These were obtained in an area with a high endemic ratio of tuberculosis, following integral biological and radiophotographic case-finding. Children who were tb contacts, belonging to the first group also included the highest proportion of allergies (48.3%) while in the other two groups the number of allergic children was similar to that of allergic children in the locality with a high endemic ratio of tuberculosis. The radiophotographic examination of adults who were contacts of tuberculous patients showed that the lowest proportion of tuberculous patients with active tuberculosis (0.4%), which was found in the third group, was still above the instantaneous prevalence of bacilliferic patients from the area with high ratio of endemic tuberculosis. In conclusion, the present technical structure of epidemiological investigations corresponds to the actual level of the endemy and no group of contacts can be excluded from the investigation. PMID- 6296973 TI - [Tuberculosis in children as an important indicator in the fight against tuberculosis]. AB - In spite of the fact that tuberculosis in children is continuously declining (in the Brasov District it fell from 197 cases during 1960-1970 to 67 between 1977 and 1979), a large number of cases are still recorded in children under the age of 3 years (from a percentage of 23 of the total number of sick children in 1969 to 44.8 in the last period). Epidemiologic studies discovered infection sources in 91% of the vaccinated children (parents represented 68% of all sources). Thus the source in the proximity, and detectable through the epidemiologic investigation exceeds the protection provided by the BCG vaccination, which results in a better protection against extra-familial infections, more rare and sporadic. It appears that in tuberculosis sources BCG vaccination alone is less efficient and a more prompt application of other preventive measures should be considered, such as early detection, rapid removal of the source, removing the children from the infective environment. Considerations should also be given to a control of the parents, especially of the father, which appears to be the source of infection twice as frequently as the mother. PMID- 6296974 TI - [Comparative evolution of the incidence of bronchopulmonary tuberculosis and bronchopulmonary cancer (data of the TB dispensary No. 1 in Galati)]. AB - Over a period of 10 years a decrease was noted in the area of the Tuberculosis Dispensary of the Galatzi District, of the incidence through tuberculosis of the adult by approximately 50%, and an increase of the pulmonary cancer incidence three fold. The incidence curves of these two diseases, which were far apart 10 years ago, are much nearer now, and it may be presumed that in a not too remote future they will cross each other. The pneumophysiology network should be prepared further to face the increasing flow of patients with non-tuberculous diseases. With this view physicians should be re-cycled, and various types of specialists should be trained for cytologic diagnosis, and the technical equipment of antituberculous hospitals should be updated. PMID- 6296975 TI - [Incidence and prevalence of chronic pulmonary tuberculosis in the Cimpulung Arges district 1972-1980]. AB - The study concerns 53 cases of chronic and hyperchronic pulmonary tuberculosis followed over a period of 9 years (1972-1980). These cases are analysed from the viewpoint of the frequency of the disease in relation to age, sex and environment, as well as from the viewpoint of the localization, morphology, and bacteriology of the lesions. References are further made to causes which lead to chronicization of pulmonary tuberculosis starting from the cases in this group. The efficiency of the strictly supervised treatment is evaluated by the dynamics of chronic pulmonary tuberculosis over the 9 year period, as well as problems related to socio-professional rehabilitation of the patients. PMID- 6296976 TI - [Risk of bronchopulmonary cancer in patients with chronic obstructive bronchopneumopathy]. AB - A total of 477 cases have been followed, presenting with chronic obstructive bronchopneumopathy, and another 151 cases with chronic bronchitis without obstructive syndrome, over a median duration of 5 years. In the first lot of patients 37 cases of bronchopulmonary cancer have been recorded over this period of time, of which 35 in 333 male patients (10.5%), and 2 in 144 women (1.4%). The annual rate of bronchopulmonary cancer was of 1.9% in males, and of 0.2% in females. In the group of patients with simple bronchitis only 3 cases of bronchopulmonary cancer have been recorded, with an annual rate of 0.4%. In male patients with chronic obstructive bronchopneumopathy the annual risk of bronchopulmonary cancer is of 1900 0/0000, 60 times greater than in the general population, where it is of 30 0/0000. A comparative analysis of the three groups: chronic simple bronchitis, chronic obstructive bronchopneumopathy and chronic obstructive bronchopneumopathy complicated with bronchopulmonary cancer shows that in the last group there are several outstanding features: a more advanced median age, intensive (heavy) smoking, a more advanced degree of chronic alcohol intoxication, frequent professional exposure, genetic deficiencies of I.A.T., prolonged corticoid therapy in the antecedents, and a higher proportion of associated diseases. In general this group is more exposed to risk factors, and has a higher risk of alteration of the defense mechanisms of the organism. However, a more detailed analysis is necessary for detecting the factors involved in the genesis of bronchopulmonary cancer in patients with chronic obstructive bronchopneumopathies. PMID- 6296977 TI - [Results of continuous treatment (3 years) with disodium cromoglycate of patients with bronchial asthma and spastic bronchitis]. AB - A group including 36 patients with bronchial asthma and 8 with spastic bronchitis have been treated with disodium cromoglycate for a period of 3 years starting with July 1-st, 1978. Eight of these patients also had allergic rhinitis, and in these ones administration of the treatment was also made by nasal application. Partial results have been published after 3 and 18 months of treatment. Of the original 44 patients 23 are still followed and the other 21 had been followed for at least 2 years, an interval which was considered as sufficient for eliminating seasonal variations of the asthmatic manifestations, independent from the treatment applied. The results at 3 years are different from those at 3 months and are relatively similar to those at 18 months. In 41.2% of the cases necessities for steroid therapy had been reduced, and in 30.3% of the cases a durable suppression of corticoid therapy was possible. With regard to bronchial dilators the doses have been diminished in 32.1% of the cases, and completely abandoned in 20.3% of the patients. A synthetic evaluation of clinical and laboratory results showed that these were positive in 68.3% of all cases. The treatment was well tolerated and no undesirable side effects were noted over the period of the treatment. PMID- 6296978 TI - [Some aspects of current medical treatment of bronchopulmonary suppurations]. AB - Due to their frequency, which is still significant in the frame of respiratory pathology, bronchopulmonary suppurations especially those of the secondary type, developing on previously existing lesions such as chronic bronchitis, bronchiectasis, bronchial asthma, bronchial carcinoma, aerial cysts, etc. rise bacteriological, therapeutical and epidemiologic difficulties for the general practitioner. An analysis is presented, of a group of 619 patients with suppurated bronchopneumopathies, hospitalized in 1980 in a department for lung diseases, of whom 116 had primary, and 503 secondary suppurations. In 177 cases (28.5%) monomorphous flora was identified, from the Ps. aeruginosa and Esch. coli strains, as well as Staphylococcus aureus. In 395 patients (64%) cultures remained sterile, and in 171 cases (27.6%) frequent fungal colonies of the Candida genus were evidenced. The medical treatment, which is sometimes difficult due to the severity and the variety of anatomo-clinical forms of the disease, includes, on an individualized basis, symptomatic therapy, anti-infectious and adjuvant therapy dominated by fluidifying drugs, by antibiotics adapted to the various microbial strains, as well as to particular conditions such as acute episodes or superinfections with anaerobic germs. There is an increased interest for re-establishing immunological balance by substitution medication or bio stimulating preparations such as mono- or polymicrobial vaccines. PMID- 6296979 TI - [Experimental studies concerning the exact determination of the etiopathogenesis of pneumoconiosis of coal miners]. AB - The effects have been studied of some pure charcoal powder, and of mixed "coal pit" powder which are representative for the basin in the Jiu Valley. A total of 48 samples have been studied as such and after addition of quartz graduated to contain 5, 10, 15, 20 and 25% quartz. The aim of the study was to determine the type of pneumoconioses induced by the various powders, to establish the role of quartz particles and the threshold at which quartz addition will impart silicogenic properties to the dust particles. A total of 300 Wistar rats have received intra-tracheal doses of an unique amount of 50 mg suspension as such and with added quartz. The rats were distributed in 12 groups. Three animals in each group were sacrificed at 6 and 12 months after instillation and histopathologic studies of the lungs were performed. Addition of 10% quartz dust to the charcoal powder, and of 15% to the "coal pit" powder imparted silicogenic properties to the samples. Both powders have masked to a certain extent the effects of the added quartz, the protective effects being exerted by minerals associated to charcoal (silicates, metallic oxides, etc.). In the mining area investigated pneumoconioses and silicosis represent two distinct pathological entities, and the role of the quartz particles in the genesis of pneumoconioses in this particular area appears to be limited. PMID- 6296980 TI - [Acute fungal pneumopathy caused by Candida and Aspergillus (case report)]. AB - A case is presented, of a patient with acute fungal pneumopathy. The evolution of the disease in this 63 years male was short and rapid, characterized by fever, dyspnea, thoracic pain, muco-purulent expectoration and intense coughing. In spite of the complex treatment applied the patient died with acute cardio respiratory failure. The radiological examination revealed micro-nodular and nodular opacities extending predominantly in the left lung. Laboratory investigations included a microbiologic study for the Koch bacillus, which was negative. The post-mortem examination revealed multiple focuses with suppurative aspects on a diffuse background of hepatization. The microscopic examination evidenced the presence in the respective focuses of abundant colonies of Candida and Aspergillus, and a hematogenic invasion with Candida and allowed to determine a diagnosis of acute pneumopathy of fungal origin as a result of a double infection with Candida and Aspergillus strains. PMID- 6296981 TI - [Eosinophilic pulmonary infiltrate (considerations of a case of Loeffler syndrome)]. PMID- 6296982 TI - [A rare case of tuberculosis caused by musculo-cutaneous inoculation]. PMID- 6296984 TI - PCI in small cell carcinoma of the lung. PMID- 6296983 TI - [Efficacy of ketoconazole in patients with recurrent paracoccidioidomycosis]. PMID- 6296985 TI - [Reconstruction of tracheobronchi in malignant disease]. PMID- 6296987 TI - [Tracheoplasty and its problems]. PMID- 6296986 TI - [Surgery of the tracheal tumor]. PMID- 6296988 TI - [Anticonvulsant action of pargyropyranone in various models of experimental epilepsy in the cat]. AB - The new anticonvulsant drug pargypiranone was studied in different models of experimental epilepsy in cat (penicillin and Premarin cortical focus, cobalt cortical focus, penicillin generalized epilepsy). The drug proved to possess a strong anticonvulsant effect upon focal electroclinical phenomena and a poor effect on penicillin generalized epilepsy. It seems probably that pargypyranone has a valid effect on partial epileptic seizures. PMID- 6296989 TI - [Primary malignant tumors of bone. Rare tumors]. PMID- 6296990 TI - [Histochemical identification of lymphocytic infiltration in the rheumatoid nodule]. AB - The authors have used histochemical techniques to identify lymphocytic infiltrations in the rheumatoid nodule. They used the technique of alpha-naphtyl acetate esterase (ANAE) for macrophages, ANAE plus Sodium Fluoride (NaF) for T lymphocytes and 5-nucleotidase for B-lymphocytes. The results of this study reveal that the lymphocytic component of the rheumatoid nodule is mostly B lymphocytes. PMID- 6296992 TI - [Giant mixed tumor in a unusual location. Apropos of a case]. PMID- 6296991 TI - [Serological studies for research on arbovirus antibodies in a human population of the Vale do Ribeira region. III. Survey in inhabitants with cases of Flavivirus Rocio encephalitis]. PMID- 6296993 TI - Bile acid metabolism in non-jaundiced patients with malignant liver tumours. AB - Bile acids were determined in the serum and urine of four non-jaundiced patients with secondary liver tumours by means of gas-liquid chromatography. In serum only the concentration of cholic and chenodeoxycholic acids were increased in two of the patients. The renal excretion of cholic and/or chenodeoxycholic acids was increased in all patients, whereas the excretion of hyocholic acid was increased in two. The renal excretion of 3 beta-hydroxy-5-cholenic acid was not increased in any of of the patients. One patient with multiple liver metastases was studied during the development of jaundice. During this period there was an increase of serum concentrations and renal excretion both of unsulphated cholic, hyocholic, and chenodeoxycholic acids and of sulphated chenodeoxycholic and 3 beta-hydroxy-5 cholenic acids. One non-jaundiced patient with a primary liver tumour, a hepatocarcinoma, had almost normal serum concentrations and renal excretion of bile acids. PMID- 6296995 TI - Hereditary myeloperoxidase deficiency: study of 12 cases. AB - 12 cases of hereditary myeloperoxidase (MPO) deficiency are reported. Histochemical stainings, lysosomal enzyme determinations, electron microscopic study of MPO and granulocytic function were performed. Family studies on 2 generations were carried out in 5 patients and histochemical stainings and biochemical lysosomal enzyme determinations were done. MPO deficiency was found to follow autosomal recessive inheritance and only rarely to have clinical effects. PMID- 6296994 TI - Immunohistochemical characterization of hepatic tissue lymphocyte subpopulations in liver disease. AB - Liver biopsies from 27 patients were studied by the indirect immunofluorescence and immunoperoxidase technique for relative distribution of B cells and T-cell subpopulations. T-cell subsets were defined for OKT4(+) (helper/inducer) and OKT8(+) (suppressor/cytotoxic) cells by using mouse hybridoma monoclonal antibodies. Patients with chronic active hepatitis and those with primary biliary cirrhosis showed marked relative increments in suppressor/cytotoxic OKT8(+) lymphocytes within hepatic lymphocytic infiltrates. Other patients with acute hepatitis or those with only reactive changes also showed a relative increment in OKT8(+) T lymphocytes within hepatic biopsy material. These findings suggest that tissue lymphocytic subpopulation increments in suppressor/cytotoxic T-cell subpopulations are commonly associated with hepatic lymphocytic infiltrates and may be related to disorders of immune regulation or expression in some of these disorders. PMID- 6296996 TI - Multiple glomus tumours. Special reference to radiological findings. AB - Glomus tumours may be either solitary or multiple. The solitary glomus is a quite common, characteristically painful lesion, frequently located under the nails, whereas multiple glomus tumours constitute a rare clinical and genetic entity which is defined by the presence of small bluish intradermal tumours, and autosomal dominant mode of inheritance. The lesions resemble clinically and histologically cavernous hemangioma, but can be differentiated by intraluminar presence of the characteristic glomus cells. Four cases of multiple glomus tumours, one sporadic and three hereditary familial, are reported, and the pedigree of two families with five generations affected by the disease are presented. Radiological investigations of multiple glomus tumours are very sparse, and their results inconsistent. Angiography, venography and thermography did not contribute to diagnosis in the present cases. Skeletal changes associated with multiple glomus tumours are discussed. PMID- 6296997 TI - Results of surgical treatment of lung metastases in children. AB - Between 1970 and 1979, 14 patients were operated on for pulmonary metastases. In 9 of these the primary tumour was a Wilms' nephroblastoma. Five out of the 14 patients are presently alive and free from disease (3 to 10 years after pulmonary resection). Primary tumours of the survivors were Wilms' tumour in 3 cases, an ovarian teratoma in one patient and a Ewing's sarcoma in one patient. Two of survivors had bilateral involvement and one had multiple metastases of one lung with pleural exudation. The disease-free interval was the same among the survivors and the non-survivors. It is apparent from the present material that some patients with a very poor prognosis as predicted from different prognostic factors can be cured after surgical excision of the metastases. The surgical procedure in children involves a very small risk and only moderate discomfort. An active surgical approach is therefore recommended. PMID- 6296998 TI - Homologous regulation of gonadotropin-releasing hormone receptors in cultured pituitary cells. AB - Specific receptors for gonadotropin-releasing hormone (GnRH) in cultured rat pituitary cells were increased by subnanomolar concentrations of GnRH agonists and decreased by high concentrations of these peptides. The antagonist [D-Phe2, Pro3, D-Phe6]GnRH did not alter GnRH binding capacity and blocked the increase in sites induced by GnRH. These findings provide direct evidence for the homologous regulation of GnRH receptors by physiological concentrations of the hypothalamic peptide, an action that could mediate the cyclical and postcastration increases in GnRH receptors and responsiveness of the pituitary gonadotrophs. PMID- 6296999 TI - Anti-inflammatory steroids without pituitary-adrenal suppression. AB - When two new steroids, methyl prednisolonate and methyl 20-dihydroprednisolonate, were applied locally their anti-inflammatory activities were nearly equivalent to those of the parent compound prednisolone in the cotton pellet granuloma bioassay. However, when these two derivatives were administered systemically, their anti-inflammatory activities were weaker than those of the parent compound. Furthermore, unlike the parent compound, these new anti-inflammatory steroids did not suppress pituitary-adrenal function or cause liver glycogen depletion in rats. PMID- 6297000 TI - Monoclonal antibody to acetylcholine receptor: cell line established from thymus of patient with Myasthenia gravis. AB - A human B cell line producing a monoclonal antibody to an antigenic determinant of acetylcholine receptors was established by cloning B cells that had been transformed in vitro by Epstein-Barr virus. The B cells were obtained from the thymus of a patient with myasthenia gravis. The antibody produced by the cell line precipitated acetylcholine receptors from denervated and innervated rat muscle and from human muscle, but did not show detectable response to the acetylcholine receptors from the electric organs of Narke japonica. The monoclonal antibody showed identical binding patterns in innervated and denervated rat muscles. Passive transfer of the monoclonal antibody into rats induced moderate muscle weakness and electromyographic changes characteristic of myasthenia gravis. PMID- 6297001 TI - Oncogene from human EJ bladder carcinoma is located on the short arm of chromosome 11. AB - The human cellular homolog of the transforming DNA sequence isolated from the bladder carcinoma cell line EJ was localized on the short arm of human chromosome 11 by Southern blot analysis of human-rodent hybrid cell DNA. This locus contains human sequences homologous to the Harvey murine sarcoma virus v-Ha-ras oncogene. PMID- 6297002 TI - 5' viral and human cellular sequences corresponding to the transforming gene of simian sarcoma virus. AB - The 5' nucleotide sequences of the transforming gene of simian sarcoma virus (v sis) and its human cellular homolog (c-sis) were compared. A short homology was found between helper virus and cellular DNA sequences at the junction of v-sis and c-sis, which may have had a role in the original recombination event leading to the generation of simian sarcoma virus. PMID- 6297003 TI - Expression of a cellular oncogene during liver regeneration. AB - The number of transcripts of the cellular oncogene ras, which is homologous to the transforming gene of Harvey sarcoma virus, increases during liver regeneration in rats. The increase in these transcripts in liver polysomal polyadenylated RNA occurs at the time of activation of DNA synthesis during the regenerative process induced by partial hepatectomy or carbon tetrachloride injury. The number of ras transcripts returns to basal levels within 72 hours. These observations show that transcription of a cellular oncogene increases in a regulated way in a nonneoplastic growth process. PMID- 6297004 TI - Rabies virus glycoprotein analogs: biosynthesis in Escherichia coli. AB - The surface of rabies virus is composed of an approximately 60,000 dalton glycoprotein, in which most of the antigenic and immunogenic determinants of the virus reside. We have constructed plasmids for the direct expression in Escherichia coli of the mature full length rabies glycoprotein gene and also for the expression of a glycoprotein gene which has been truncated to exclude the coding region for a hydrophobic, possibly transmembrane, domain of the protein. Escherichia coli harboring the plasmids synthesize analog proteins which conform by several biochemical and antigenic criteria to rabies glycoprotein. PMID- 6297005 TI - Multiple point mutations affecting the simian virus 40 enhancer. AB - Enhancers, or activators, dramatically increase the transcriptional activity of certain eukaryotic genes. A series of multiple point mutations affecting the simian virus 40 (SV40) enhancer-activator region were generated in order to define the nucleotide sequence required for this function. Three independent assays provided information leading to the identification of nucleotides essential for enhancer function. One class leads to a decrease in gene expression, while the second completely abolishes functional activity. One critical replacement appears to be the first G (guanine) in a sequence TGGAAAG (T, thymine, A, adenine) located in the 5' region of the 72 base-pair repeat of SV40. Comparison of this sequence with nucleotide sequences in other known enhancers leads to the identification of potential related core elements. PMID- 6297006 TI - Monoclonal antibodies for diagnosis of infectious diseases in humans. PMID- 6297007 TI - Prospects in plant genetic engineering. AB - The functional expression of a novel gene in a genetically engineered plant has not yet been reported. One major barrier in movement toward this goal is our limited understanding of the molecular bases of gene expression. Attempts to establish genetic engineering as a practical facet of plant breeding are also complicated by the fact that genes for most important plant characteristics have not yet been identified. However, the benefits to be gained from all aspects of plant improvement are stimulating research into both the development of plant transformation technology and the isolation and characterization of genes responsible for valuable traits. As scientists develop greater knowledge of plant molecular genetics, we can expect to see practical applications in such diverse areas as improvement of plant nutritional quality, decreases in fertilization requirements, and increases in resistance to environmental stresses and pathogens. PMID- 6297008 TI - Actions of estrogens and progestins on nerve cells. AB - Estrogens and progestins alter electrical and chemical features of nerve cells, particularly in hypothalamus. Temporally, these events follow nuclear receptor occupation by these steroids, although not all effects have been proved to depend on translocation of receptors to the nucleus. Narrowing studies to focus on particular medial hypothalamic cells has been useful for understanding some of the actions of these steroids in brain. The variety of morphological, chemical, and electrical effects allow for a multiplicity in the cellular functions controlled by these hormones. PMID- 6297009 TI - Intracellular calcium measurements with arsenazo III during cyclic AMP injections into molluscan neurons. AB - Injections of cyclic adenosine monophosphate into molluscan neurons often produce a transient membrane depolarization. By using the calcium indicator dye arsenazo III, it was found that cyclic nucleotide injections into neurons of Archidoris montereyensis resulted in elevation of internal calcium concentrations. However, this was demonstrated to be a secondary consequence of an induced increase in membrane sodium permeability, and not due to any direct effect of cyclic adenosine monophosphate on cellular calcium influx or internal calcium regulating processes. PMID- 6297010 TI - Ammonium chloride prevents lytic growth of reovirus and helps to establish persistent infection in mouse L cells. AB - Ammonium chloride, a lysosomotropic agent that raises intralysosomal pH, reduces the yield of reovirus during infection of mouse L cells. Subsequent removal of ammonium chloride results in the rapid establishment of a persistent infection. PMID- 6297011 TI - [Complications of antitumor and antileukemia chemotherapy. 3 (conclusion)]. AB - Among the neurological side-effects, peripheral neuropathy is a result of therapy with vindesine and above all vincristine. Although in most cases it is responsible only for paresthesias, it may cause extensive paralysis and requires that the drug be discontinued. These drugs may also affect the neurovegetative system. Ototoxicity may be seen with cis-platinum and vigilance disturbances with L-asparaginase. Genetic consequences are mainly due to alkylating agents. These agents almost constantly impair male and female fertility but recovery is possible. Libido is also affected with the attendant psychological consequences. The offspring of patients previously treated by chemotherapeutic agents are normal. Development of secondary carcinoma or leukemia is currently a major concern. Secondary malignant disease may develop after the treatment of any cancer, especially if radiotherapy was associated with alkylating agents. Leukemias are of the acute myeloid type and usually follow a preleukemic phase. A table summarizes the main toxicities of the most usual drugs. PMID- 6297012 TI - [Singh femoral index in vertebral osteoporosis]. AB - In 48 patients with osteoporosis and at least one vertebral fracture, the Singh vertebral index, which assesses the femoral trabeculation, was compared to the Meunier vertebral radiologic index, which is determined according to the number and severity of vertebral deformations. The femoral index was abnormal in all patients. However, the threshold level, set by Singh at Stage IV, which supposedly discriminates between subjects with and without osteoporosis could not be confirmed as one-third of the patients with at least one vertebral fracture were Stage V. A statistically significant correlation was found between the femoral index and the vertebral radiological index. PMID- 6297013 TI - [Cutaneous lesions in human African trypanosomiasis]. AB - We report the case histories of 12 patients infected by Trypanosoma gambiense seen between 1967 and 1979. The course of the disease follows two stages, with the lymphatic and hematologic stage preceding invasion of the central nervous system. The following cutaneous lesions were recorded: 5 patients had trypanomas or chancres which are inflammatory nodules on uncovered areas; 5 patients showed trypanosomal skin rashes with erythematous, infiltrated, transient and recurring trypanids; 5 patients had soft transient edema, affecting mainly the face; 3 patients experienced pruritus, which was usually mild and limited to small areas; 3 patients had cutaneous lesions induced by melarsoprol. Thus dermatologic features are common in human African trypanosomiasis; they were found in 75% of the patients in our department. PMID- 6297014 TI - [Value of cineradiography in the study of Budd-Chiari syndrome. Value of various angiographic technics. Apropos of 8 cases]. AB - Eight cases of surgically proven Budd Chiari disease, have been explored pre- operatively by cine-angiography. This procedure is very safe and simple, and allows a dynamic study of the intra- and extra-hepatic pathways. The authors are reviewing the different modalities of radiologic evaluation of Budd-Chiari disease, including inferior vena cava phlebography, opacification of supra hepatics veins, and coeliac arteriography. They propose a flow sheet in the radiological work-up of these patients. PMID- 6297015 TI - [Efficacy and tolerability of single-dose rosoxacin in the treatment of acute uncomplicated gonorrhea]. AB - 61 patients, male and female, with uncomplicated acute gonorrhea were given a single dose of 300 mg rosoxacin (2 capsules). 52 patients only completed clinical and laboratory controls. The drop-out rate for follow-up examinations was 14.8%. 98% of patients were cured. One patient failed to respond to treatment, and six had reinfection seven days later at follow-up control. Three of these were again treated with rosoxacin and cured. Among the side-effects, which were recorded in 24.4% of patients, dizziness was most prevalent (18.1%). Rosoxacin is particularly valuable in gonorrhea, given the single oral dose, the absence of resistance and the high percentage of efficacy. PMID- 6297016 TI - [Cytogenetic aspects of diffuse scleroderma. Structural anomalies and sister chromatid exchanges]. AB - Structural chromosome anomalies (1 477 cells examined) and sister chromatid exchanges after two replication cycles with BrdU (771 cells studied) were evaluated in 12 patients with diffuse scleroderma and having received no recent or important irradiation. The increase of structural anomalies, chromatidic as well as chromosomal, is always low, inconstant and cannot be considered as having a diagnostic value. Increase of sister chromatid exchanges could be a more sensitive method of investigation. In particular, it is not influenced by low doses of diagnostic X-rays. PMID- 6297017 TI - [Tiapride in disorders of behaviour in the elderly]. AB - Our experience with tiapride in 30 elderly inpatients with disorders of behaviour is reported. In a daily dosage of 50 to 600 mg, the drug was effective on sleep disorders, agitation, irritability and aggressiveness. Tolerance was excellent. PMID- 6297018 TI - [Chronic renal failure and lithiasis]. AB - After analyzing 39 cases, the authors go on to describe the virtues of surgical treatment of non-uric lithiasis after correct evaluation (lateral urography, CT scanning, retrograde ureteropyelography). The operative techniques draw considerable benefits from intraoperative radiography and, when necessary, the clamping of the renal pedicle under pharmacological protection. Surgical management offers the only possible means of preventing aggravation of the renal failure. PMID- 6297019 TI - [Cerebral tomodensitometry in neuropediatrics]. AB - CAT scan is now the primary diagnostic procedure for intracranial lesions in children. It is particularly well-suited for investigating intracranial hypertension, tumors and other space-occupying lesions, hydrocephalus, and malformations. Because it is non-invasive, this technique tends to be unduly performed as a screening procedure, with a resulting oversaturation of neuroradiological facilities. CAT scan should not be used as a routine examination, but only in well-selected cases. Careful clinical assessment and a good knowledge of pediatric neurology are still essential. New techniques are now becoming available and neuroradiologists will be able to select the best investigation for each particular case. PMID- 6297020 TI - [Spontaneous hemarthrosis in adolescents and adults, excluding hemophilia]. AB - The various etiologies of spontaneous hemarthrosis in adolescents and adults are reviewed: they include systemic diseases and local or regional disorders of the bones or joints. Among systemic diseases, the two main causes are coagulation disorders and hemoglobinopathies. Coagulation disorders may be either acquired (leukemia, thrombopenia, and hypoprothrombinemia induced by anticoagulant drugs with hemarthrosis being one of the major complications) or inherited (hemophilia which is not considered here, von Willebrand disease, and congenital thrombopathies). Hemoglobinopathies, particularly sickle-cell disease, are responsible for hemarthrosis in a few patients. Among local or regional disorders of the bones or joints, tumors such as hemangioma or synovial sarcoma are uncommon causes. Hemarthrosis is the main feature of pigmented villonodular synovitis. Hemarthrosis may occur in degenerative and metabolic diseases: while it is extremely rare in arthritis, it is frequently encountered in articular chondrocalcinosis which is the first diagnosis to consider when hemarthrosis occurs in an elderly patient. The search for an etiology, which is often difficult, should include a review of prior illnesses, a study of coagulation, and local clinical, radiological and biological investigations, with a study of the synovial fluid; in some instances, arthroscopy, synovial biopsy and even surgical exploration are required. Management includes rest, analgesics, antiinflammatory drugs and, above all, arthrocentesis which is essential for the prevention of articular damage and functional sequellae. Specific therapy is dependent on the etiology. In recurrent hemarthrosis, isotopic synoviorthesis may ensure lasting resolution of the effusion. PMID- 6297021 TI - [4 cases of Mycoplasma pneumoniae pneumonia]. AB - The authors report four cases of Mycoplasma pneumoniae pneumonia which occurred towards the end of 1981, concurrently with an intensification of this infection. Three patients had severe respiratory distress and one required ventilatory assistance. The features shown on chest roentgenograms are discussed. All four patients had been treated initially with penicillin or cephalosporin. This report is evidence that patients with pneumonia should be given a macrolid as the first treatment. PMID- 6297022 TI - [Fortuitously discovered gallbladder lithiasis and surgery]. AB - When confronted with the fortuitous discovery of cholelithiasis, a simultaneous operation is advisible in view of the drawbacks of performing a second operation and the risks of postoperative cholecystitis. Analysis of the author's results and those reported in the literature shows only a slightly increased morbidity rate and a similar mortality rate. In high-risk patients, and if the cholecystectomy appears difficult, the removal of the stones followed by a cholecystostomy is less aggressive and faster. PMID- 6297023 TI - [Treatment of menopausal hot flushes with a nonhormonal medication, veralipride]. AB - The value of veralipride in menopausal disorders is no longer questionable and has been documented by a number of studies. The purpose of this paper is to study the clinical effectiveness and to look for the biological consequences of three months therapy with veralipride. To achieve this, a thorough biological study (hematology, glucids, lipids, hepatic and renal function) was done in 21 patients with a menopausal syndrome before and after taking veralipride for three months. Clinical effectiveness was excellent with 80% satisfactory results. No signs of intolerance or adverse side-effects were seen. We recorded no significant changes in biological parameters. No disturbances of renal or hepatic function were detected, either in individual assessments or in the overall statistical analysis of the study group. Because it is well tolerated, effective, and devoid of adverse metabolic side-effects, we conclude that veralipride is the best non hormonal treatment for menopausal disorders presently available. PMID- 6297025 TI - [Aphthous hepatic lesions and Behcet syndrome]. AB - The case of a twenty-one-year-old man who was hospitalized for unexpected bilateral phlebitis of the lower limbs is reported. Clinical features suggested a systemic disease, especially Behcet disease although the classical triad was lacking. Laparoscopy showed two lesions identical to canker sores on the hepatic capsule. There were no biological hepatic disorders and histological examination of a needle biopsy specimen was normal. This clinical report is additional evidence in support of the idea that Behcet is syndrome is a systemic disease. PMID- 6297024 TI - [Adenoma and focal nodular hyperplasia of the liver. Value of radiological tests. Apropos of 15 cases in adults]. AB - The authors present the results of the radiological explorations in 15 cases of benign liver tumors. Ultrasound and/or computed tomography can usually confirm the presence of a hepatic tumor. Arteriography combined with the clinical information suggested the diagnosis. The authors stress the role of the radiological investigations for the follow-up of these tumors. PMID- 6297026 TI - [Gastrostomy lasting 59 years]. AB - We report on an unusual observation of a woman who lived with a gastrostomy for fifty-nine years. Gastrostomy was performed at age twenty-six after esophageal burn. The patient died at age eighty-five. Digestive functions were normal. Calorie intake was below normal requirements. Diet was monotonous and unbalanced with protein deficiency. However, body weight and plasma protein levels were normal. Psychological and gustative behaviours are discussed. PMID- 6297027 TI - [A case of pseudotumorous pulmonary cryptococcosis]. AB - A case of pseudotumorous pulmonary torulosis (or toruloma) clinically suggestive of protracted pneumopathy is reported. The main pathological, clinical and histological forms of this mycotic infection are recalled. The pathologist plays an essential part in the diagnosis of toruloma since the fungus can be demonstrated in bronchial biopsy specimens, bronchial aspirates and sputum. PMID- 6297028 TI - [Craniofacial dysmorphism with flexion of the fingers. Marden-Walker syndrome?]. AB - The Marden-Walker syndrome was first described in 1966. The main features are microcephaly, peculiar facies due to blepharophimosis, micrognathia, low-set ears, joint contractures, muscular hypotonia, growth failure, and developmental delay. We report the case of a child presenting with almost all of these features, but with muscular hypertonia. Differential diagnosis includes Schwartz Jampel syndrome. Pathogenesis is unknown. PMID- 6297030 TI - [Functional interpretation of villous atrophy of the small intestine]. AB - The villosities of the small intestine form a special structure (absorbing enterocytes, functional vascularisation of mucosa), manifestly adapted to the working of the absorption function. As a reciprocal, we propose the hypothesis that the maintenance of villous structures depends on the use of the absorption function and we have classified villous atrophies in three groups, according to their mechanism: by insufficiency of the absorbing enterocytary coating, alteration of the connective tissue and especially of functional vascularisation of mucosa, insufficient absorption (functional regression). Though based on a largely conjectural theoretical conception, this classification corresponds, in fact, to particular aspects of the mucosa as to organization of the connective tissue and especially to maturation of the enterocytary series: it should therefore help pathologists to add to the static appreciation of the hight of villosities and the abondancy of plasmocytes, an appreciation of anatomoclinical syndromes or, at least, of unquestionable lesional mechanism. PMID- 6297029 TI - [Muscular involvement in adult Still's disease. Apropos of 5 new cases]. AB - Still disease was first described in adults ten years ago. Muscular involvement was already mentioned in the first reports. However, it's incidence is underestimated as it is often mistaken for arthralgia. Diagnosis of muscular involvement is suggested by clinical features and established by biological, electromyographical, and histological investigations. We describe five new cases of adult-onset Still disease with muscular involvement. All five cases followed the usual favorable course. PMID- 6297031 TI - [Advances in the physiopathology of the platelet membrane]. PMID- 6297032 TI - [Insomnia therapy and withdrawal of hypnotics]. AB - The aim of this study is to evaluate the efficiency of a treatment prescribed, in the course of an hospital consultation for sleep pathology, to patients suffering from chronic insomnia not improved by longstanding and sustained medication with hypnotic drugs. The basis of the treatment is a progressive but total withdrawal of hypnotics in so far taken regularly. The withdrawal of hypnotics was prescribed to 79 patients: 33 aged 17 to 39 years (group 1, mean age 30) and 46 aged 40 to 70 years (group 2, mean age 51). 41 showed primary psychophysiological insomnia and 28 showed insomnia associated with psychiatric disorders. In patients of group 1, the average durations were 8 years for insomnia and 3 years for sustained hypnotic use; these durations were 15 and 5 years respectively in patients of group 2. Hypnotic drug withdrawal was achieved without placebos in 3 months in group 1 patients and 5 months in group 2 patients. 65 patients completely stopped the continual use of hypnotics. Subjective improvement of insomnia was reported by 51 of these patients (as well as by 6 patients who were given simultaneous antidepressant therapy). 16 of the 51 improved patients have resorted to hypnotics occasionally (at intervals of 10 days or more). After complete withdrawal, patients went on consulting for various lengths of time: 5 months average for group 1, 14 months average for group 2. This study of a fairly large group of insomniacs shows the frequent ineffectiveness of a sustained use of currently available hypnotics. It also shows that two times out of three the complete stop of sustained hypnotic medication proved beneficial to the patient. PMID- 6297033 TI - [The child as a symptom of his parents]. AB - The child psychiatrist is often consulted for symptoms which apparently belong to the child but, in fact, betray a much broader problem. This is substantiated by three clinical observations. The child's symptoms may be the only manifestation of a pathological family organization. The problem often originates in the parents' relationship: the parents can stay together only if the unconscious conflict between them remains hidden; however, the repressed pulsions remain active and provide the child with a basis for building a symptomatic complex which, as for neurotic arrangements, is a compromise between expression and repression of unconscious pulsions. In other cases, the child's symptoms represent an actualization or a revival of the parents' repressed phantasms. Thus, the child becomes a symptom of a pathological situation which affects the whole family. The child's experiences during the first years of life are often found to have been deficient. Inferiority of one of the parents, usually the father, compared to the other, is often noted. In all cases, the therapeutic approach requires special caution. PMID- 6297034 TI - [Difficulties and pitfalls in the approach to psychosomatic diseases]. AB - The specific particularities, difficulties and, in some instances, pitfalls which are attendant to the approach of "psychosomatic" patients may be a source of therapeutic satisfactions for the practitioner but also of feelings of uneasiness or helplessness. The existence of a psychosomatic relationship, i.e. of evidence which connects, at least in part, a somatic disorder to the psychological context in which it occurs, and the characteristics of the psychosomatic mechanism, i.e. of the various modalities through which psychological distress is converted into physical suffering (physical behaviour and attitudes, conversion hysteria, functional disorders, so-called psychosomatic diseases) are the two questions to be addressed as soon as diagnosis is considered. The difficulties and pitfalls are mostly met during management; the role of the physical practitioner is underscored; some utilizations of psychotropic agents as well as some of the physicians' attitudes and phrases are discussed; the value of methods involving the body (relaxation, etc.) is recalled. PMID- 6297035 TI - [Current concepts of hospitalization in pediatric psychiatry]. AB - Although it remains valuable for evaluating the patient and planning pharmacotherapy, hospitalization in a psychiatric department is no longer restricted to these purposes. Hospitalization enables separation of the child or adolescent from his usual environment during periods of acute conflict. It is aimed at modifying and overcoming the conflicts between family members which are underlying in the child's or parent's request for support. Hospitalization can become an essential part of the child's treatment if it is carefully planned and harmonized with the other curative and preventive methods. PMID- 6297036 TI - [Psychiatric emergencies and family crisis. A possible systemic approach at a general hospital]. AB - Psychiatric emergencies concern not only sectorized psychiatric organizations but general hospitals as well, especially in large cities. No set-ups for long-term management are available in general hospitals and means for short-term intermittent action should be evolved. Family crisis may be responsible for psychiatric emergencies as they may lead to decompensation due to the breaking-up of familial relationships. When confronted with such situations, practitioners must of course face the emergency by using various specific chemotherapeutic agents or by hospitalizing the patient. However, practitioners should also concern themselves with acting upon the whole family in order to help them to develop solutions to the crisis. The modalities of this global approach of psychiatric disorders are delineated in the family crisis therapeutic technique. PMID- 6297037 TI - [Role of specialized care units in the psychiatric approach of very young children]. AB - Among the various means of answering psychiatric problems in young children, specialized care units for the very young, such as the one set up at psychiatric department for children and adolescents at the Salpetriere Hospital, appear to be helpful in some situations which are psychologically highly dangerous as well as life-threatening. The modalities of the reception and therapy of the child and its family are described and discussed. It seems that this type of hospitalization may contribute to the resuming of dynamic developing processes in the child and of positive relationships between child and family, particularly the mother. PMID- 6297038 TI - [Do psychiatric emergencies exist in children and adolescents?]. AB - From our experience in the pedopsychiatric emergency unit at the Salpetriere Hospital, we discuss the concept of psychiatric emergency in children and adolescents. In 90% of cases, patients are adolescents aged 14 to 18 years. The acting out and the ensuing emergency consultation seem to be a propitious time for establishing a relationship with the adolescent, meeting his difficulties from a psychological rather than affective standpoint and instituting an exchange with the family. PMID- 6297039 TI - [Organization of the department of psychotherapy for children at the Salpetriere Hospital in Paris (department of Professor D. J. (Duche)]. AB - The staff of the department of psychotherapy at the Salpetriere Hospital (Department of Prof. Duche) includes five psychoanalysts who are in charge of the consultations and treatment of hospitalized children, out-patients and parents. Individual psychoanalytical psychotherapy and psychodrama are the methods used. For adolescents and psychotic children, psychotherapy is carried out in a more flexible way. Special attention is given to the outset of therapy and to the support provided to parents. Two groups for the training of child therapists are ongoing. PMID- 6297040 TI - [Tiapride in acute stages of drug addiction]. AB - We studied the effect of tiapride on alcoholic and drug-dependent inpatients. In both the treatment was initiated at a time of acute intoxication. Tiapride was perfectly tolerated and we recorded none of the side effects seen with neuroleptics. The method of administration (continuous intravenous infusion) together with the mild sedative effect prompted the regressive response which is a necessary step before the subsequent reconstructive phase. The unquestionable though not overpowering sedative effect gives the patient a feeling of comfort and allows him to participate actively in the treatment. The second original characteristic of tiapride is that it's anxiolytic effect is mild, with the drawback of leaving anxiety and insomnia unalleviated. But this treatment has a novel advantage in that it does not induce any drug dependency, even secondarily. PMID- 6297041 TI - [Current aspects of acute renal failure]. AB - The retrospective review of 115 case-histories of patients with acute renal failure (ARF) seen over the last two years showed that etiologies were distributed as follows: acute tubular necrosis in 65% of cases, urinary tract obstruction in 16%, acute glomerulonephritis in 3,5%, acute interstitial nephritis (AIN) in 8% and acute microvascular nephropathy in 3,5%. The diagnostic value of renal biopsy in ARF is discussed. In spite of recent advances in the treatment of ARF, the mortality rate remains as high as 48%. This is mainly due to current etiologic circumstances, to the age of the patients and to the complications of ARF, with infectious complications being the most serious. Urea nitrogen accumulation is not a poor prognosis factor. Furosemide in high doses does not alter the prognosis but reduces the total number of dialysis indications (81% in 1970, 60% in 1980), the number of dialysis sessions per patient (1 only in 62% of patients), and the duration of the ARF episode (mean duration: 10,7 days). PMID- 6297042 TI - [Acute perihepatitis of genital origin (Fitz Hugh-Curtis syndrome): 4 case reports]. PMID- 6297043 TI - [Leukemic priapism. Physiopathology and therapeutic approach. Review of the literature apropos of 2 cases]. PMID- 6297044 TI - [Antihypertensive therapy with a single daily dose of sotalol]. PMID- 6297045 TI - [Herpes gestationis: immunologic findings]. AB - The treatment of herpes gestationis has been considerably changed in recent years by progress in electron microscopy and immunology. Herpes gestationis, a rare polymorphic vesiculo-bullous pruriginous dermatosis, occurs during pregnancy or the post-partum period. It evolves by acute periods of several weeks, and relapses during later pregnancies or on taking oestro-progestagens. Histologically, the bulla is situated at the dermo-epidermic junction. The epidermal oedema is both intra and extra-cellular. The ultrastructural studies show the importance of the oedema and the changes in the plasma membrane of the basal cells in the onset of the vesicles and the bullae. The immunological inquiry gives diagnostic certainly when one discovers under indirect immuno fluorescence, a specific serum factor, the herpes gestationis factor. These ultrastructural and immunological data suggest a relationship between herpes gestationis and bullous pemphigus (Lever's disease). In treatment, the possible pathogenic role of prolactin suggests the use of bromocriptin during the postpartum period. The efficacy and innocuity of vitamin B6 make corticosteroids rarely necessary. PMID- 6297046 TI - [Value of an novel molecule, veralipride, in the treatment of menopausal disorders]. AB - Veralipride in a daily dosage of 100 mg given twenty days each month was tested in 17 patients with hot flushes. Hot flushes and attendant psycho-functional symptoms were significantly improved in ten cases. With the exception of drowsiness in a few patients (improved by evening dosing) clinical tolerance was good. No modifications in biologic parameters were recorded. We believe that veralipride should be tried in menopausal psycho-functional disorders whenever hormonal replacement therapy is contraindicated. PMID- 6297047 TI - [Kawasaki syndrome in a young adult. Dermatologic and immunologic study]. PMID- 6297048 TI - [Bladder sphincter disorders in multiple sclerosis: symptomatology and evolution. 100 cases]. AB - The semiology and evolution of vesicosphincteral disorders have been studied in a series of 100 cases of multiple sclerosis. All the patients underwent a complete checkup and were followed over a period of 6 years. Mictional problems appear within the first five years in 60 p. cent of cases. When they occur later on, they tend to follow the evolution of the disease on their own. There exist regular correlations between the gravity of the motor and sphincteral handicap, and between the size of the postmictional residue (over 20 p. cent of the miction), the time elapsing since the onset of the mictional disorders, and the appearance of uronephrological complications. PMID- 6297049 TI - [Iron-deficiency anemia. Hematologist's viewpoint]. AB - Enlarged spleen, fever, increased susceptibility to infections, and thrombocytosis, are manifestations of iron deficiency which are relatively specific of pediatric patients. Iron deficiency anemia is part of everyday pediatrics. Patients are referred to the hematologist in the following situations: 1) Therapy is ineffective for one of the following reasons: the hypochromic anemia is not caused by iron deficiency (hemoglobinopathies); iron is less efficiently used because of transferrin deficiency or infectious, inflammatory or cancerous disease; iron therapy is inadequate either because of insufficient dosage or of suboptimal duration. 2) A relapse occurs in spite of adequate therapy. Before investigating the digestive tract, abnormal hemostasis. Osler-Weber-Rendu syndrome and pulmonary hemosiderosis should be considered. 3) Iron deficiency anemia is less common in adolescents. This condition, known as chlorosis, results mainly from increased needs, unbalanced diet, and onset of menses. In some cases no explanation is found but iron therapy leads to recovery. 4) Difficult problems arise in patients with complex anemias: iron deficiency with folic acid or vitamin B12 deficiency; hyposideremia complicating one of the hemoglobinopathies. PMID- 6297050 TI - [Folic acid deficiency in pediatrics]. PMID- 6297051 TI - [Endorphins in psychiatry]. PMID- 6297052 TI - [Neuroendocrinology of depressive disorders]. PMID- 6297053 TI - [Pharmacokinetics of psychotropic drugs: importance in psychiatric practice]. AB - The clinical importance of pharmacokinetic studies of psychotropic drugs is described. First, definitions of the main pharmacokinetic parameters which allow accurate evaluation of the fate of a drug in the organism are given. Secondly, two important points are considered: the variability of drug plasma concentrations from one patient to another with identical doses makes "standard dosages" worthless; current data evidences correlations between the therapeutic effect of a drug and it's plasma concentrations. Clinical implications of the above points are as follows: adjustment and prediction of dosages; definition of rational precepts of prescription; definition of principles for monitoring treatment. These implications are exemplified by clinical cases reported in the medical literature or seen in the department of psychiatry at the university hospital in Besancon. PMID- 6297054 TI - [Value of quantitative electroencephalography in psychiatry]. AB - Computerized quantitative EEG processes the signal in four sequential steps: 1) EEG sampling with rejection of artefacts; 2) Fourier analysis (power spectrum according to frequency); 3) data reduction with computation of spectral parameters in five-minute recordings on at least four EEG channels (mean frequencies, mean amplitudes in microvolts and %, with standard deviations for delta, theta, alpha and beta frequency bands); 4) data base and multivariate statistical analyses. Five main results are obtained by computerized quantitative EEG analysis in psychiatry and clinical pharmacology: 1) longitudinal studies of patients before and after treatment; 2) transversal studies of homogeneous EEG groups (discrimination between tracings of residual-type or paranoid-type schizophrenic patients) comparatively with control groups (high and low alpha subjects); 3) study of psychotropic drugs: profiles, prediction of action, dose effect curves, pharmacokinetic studies, biodisponibility, correlations with plasmatic levels and behavioural or clinical psychiatric rating scales; 4) typologic classification and discrimination by multivariate analysis, studies of functional activities: functional interhemispheric lateralisation of amplitudes and its variations during the course of psychotic processes; 5) new techniques: computerized topo-electroencephalography (CT-EEG, BEAM technique) based on 16 EEG recordings, comparison with other imaging techniques. PMID- 6297055 TI - [Sultopride and agitation]. AB - Sultopride, presented in this paper, was tried in five patients with psychomotor agitation and aggressiveness but with a variable underlying structure, from severe neurosis (borderline?) to chronic decompensated psychosis. These manifestations had failed to respond to the other neuroleptics used. Results with sultopride were excellent. This drug deserves a good position among sedative agents with no significant effect on vigilance used in aggressiveness. PMID- 6297056 TI - [Sultopride treatment of a chronic psychotic child with predominantly maniacal manifestations]. PMID- 6297058 TI - [Hepatic amebiasis. A review of 86 cases seen in Paris]. PMID- 6297057 TI - [Results of surgical treatment of Basedow's disease]. AB - Diffuse hyperthyroidism occurred in 88 patients (82 women and 6 men), aged 14 to 74 years, and was due to Graves disease in 65 cases and heteromultinodular goiter in 23 cases. After medical preparation, the surgical procedure was subtotal thyroidectomy for Graves disease and total unilateral with partial controlateral lobectomy for heteromultinodular goiter. The immediate postoperative course was uneventful except for two transient acute episodes of thyrotoxicosis. One to eight years after surgery, 53 patients are euthyroid, 34 are hypothyroid and one has recurrent hyperthyroidism. A TRH test was done in each of the 53 euthyroid patients. Three kinds of responses were demonstrated: a normal response in 22 cases, evidencing true euthyroidism; an excessive response in 24 cases, reflecting partial compensated hypothyroidism; an insufficient response in 7 cases. PMID- 6297059 TI - [Estrogenic paradox: treatment of ulcerated or atrophic scirrhous carcinoma of the breast in elderly women. A method not to be forgotten]. AB - The authors report their experience in the treatment of advanced scirrhous carcinoma of the breast with estrogens only at the Curie Institute. Patients were elderly women in whom other forms of treatment were contraindicated because of local tumor extension or a poor general condition related to age. 32.5% of the patients responded favorably (out of a total of 40 patients). Mean duration of treatment was 21.5 months. Dosage varied according to efficacy and side-effects. Both dienestrol and diethylstilbestrol were used with no preference for one or the other. Serious side-effects were relatively infrequent and minor side-effects either resolved or improved significantly when dosage was reduced or treatment discontinued. Considering the present widespread use of antiestrogens and the systematic detection and dosage of intratumoral hormone receptors, the authors stress the advantages of this seemingly paradoxical effect of estrogens and underline the need for comparative studies. PMID- 6297060 TI - [Increased activity of adenine phosphoribosyl transferase in a child trisomic for 16q22.2 to 16qter due to malsegregation of a t(16;21) (q22.2;q22;2)pat]. AB - Two cousins with trisomy for the distal third of 16q due to a familial translocation, t(16;21)(q22.2;q22.2), are reported. The APRT gene locus could be assigned to 16q22.2 to 16qter. The phenotypic similarities of the first patient, who had trisomy 16q22.3 to 16qter and monosomy 21q22.3, to six other patients with partial trisomy 16q reported in literature allows the delineation of a syndrome due to trisomy 16qter. In the second patient, with trisomy 16q22.3 to 16qter and trisomy 21pter to 21q22.2, similar features are present, but in association with the classical symptoms of trisomy 21. PMID- 6297061 TI - [Clinical experience with tiapride in the treatment of extrapyramidal syndromes]. AB - Tiapride was given at an average dosage of 300-900 mg per day, intramuscularly for the first three days, then by oral route, to 62 patients with abnormal movements of various types. The best responses were recorded in chorea and bucco lingua-facial dyskinesia as well as in dyskinetic movements induced by levo-dopa in Parkinson patients. Clinical and biological tolerance of tiapride was excellent, with only one case of drowsiness and two cases of galactorrhea. PMID- 6297062 TI - [Gastric pseudolymphomas. Apropos of 3 cases. Review of the literature]. AB - The authors report three cases of gastric pseudolymphoma, before going on to review the literature on the subject. The pseudolymphoma has only recently been identified in relation to other primitive gastric lymphoid growths, on the basis of histological and evolutional criteria, which show it to be benign. It raises two problems. Firstly, a practical problem of diagnosis, as it often visualizes radiologically and endoscopically as a malignant tumor, and this diagnostic uncertainty persists, even after endoscopic biopsy. This necessarily implies surgical management of the case, normally leading to a clear diagnosis based on study of the gastrectomy specimen. Histologically, a growth of this nature consists of a very large lymphoid hyperplasia, often of an organoid type, strictly confined to the mucosa, which, incidentally, shows no real recent or ancient ulceration. Cytologically, this lymphoid infiltrate is benign and the neighbouring nodes are always normal. Point by point, therefore, these characteristics distinguish the pseudolymphoma from the real lymphoma and the lymphoid hyperplasia associated with ulcers. Fluorescence microscopic study of the intracytoplasmic immunoglobulin, which was carried out in the cases treated by the authors, and also in another recent case reported in the literature, shows a polyclonal type immunofluorescence. The pseudolymphoma is therefore, a benign form of lymphomatosis without ulceration. The second problem concerns the real nature of the lesion: a local immunosuppressive pathology? An atypical lymphoid presarcomatous hyperplasia? In order to support these hypotheses, fresh tissue lymphocyte typing tests covering the entire field of digestive lymph pathology should be carried out to complete the intracytoplasmic immunofluorescent study of the immunologically competent cells. PMID- 6297063 TI - [A clinical trial of the effectiveness and tolerance of bepridil in unstable angina pectoris]. AB - Bepridil, a new anti-anginal drug, was given in a daily dosage of 400 to 600 mg to twenty patients with unstable angina pectoris. The trial was designed to evaluate the effectiveness of bepridil in this indication and to determine the optimal dosage. A positive response evidenced by the arrest of progression towards infarction was recorded in 17 patients. Tolerance was satisfactory in 19 cases: in one diabetic patient under insulin symptoms of hypoglycemia resolved once insulin was discontinued. With the dosages used the effectiveness of bepridil in unstable angina pectoris is similar to that recorded in effort angina with 300 mg per day. PMID- 6297064 TI - [Colonic localization of pseudopolypoid type disclosing Recklinghausen's neurofibromatosis]. AB - A case of colonic Recklinghausen neurofibromatosis found in a forty-two-year-old woman is presented. Only the colon was affected. The first symptom was rectal bleeding. Widespread polypoid lesions involving the rectum, sigmoid, left colon and part of the transverse colon were demonstrated upon barium enema and rectosigmoidoscopic examinations. As demonstrated by a review of the medical literature, colonic involvement in Recklinghausen disease is very infrequent and forms restricted to the colon are exceptional. The pathologic findings and the clinical, radiological and endoscopic features in colonic neurofibromatosis are described. It seems that total colectomy should be advocated. Our patient underwent total colectomy ten years ago and has experienced neither recurrence nor other manifestations of Recklinghausen disease. PMID- 6297066 TI - [Immunology of multiple sclerosis]. AB - Multiple sclerosis is an inflammatory disease which involves only the central nervous system. The intrathecal immunologic response consists mainly of local antibody production by immunocytes located in the central nervous system. The role of such immunocytes is obscure. Investigations of the immunological status of multiple sclerosis patients have shown increases in some antiviral antibodies, a cytotoxic cellular response against central nervous system cells, and abnormal proportions of lymphocyte subpopulations in the peripheral blood. Impairment of immunological control mechanisms seems highly probable. PMID- 6297065 TI - [Free radicals in medicine and biology]. AB - Free radicals have long been well known by physicists but have only interested biologists since 1969 when Fridovich showed that O2 was produced during an enzymatic oxidation. O2 and related radicals are highly toxic. This implies that, in all aerobic cells, mechanisms exist which inactivate free radicals as soon as they are produced by oxidative metabolism. O2 radicals are eliminated by a family of enzymes called superoxide dismutases (SOD). These SOD are present in the cytosol (CuSOD) and in the mitochondria (MnSOD). Overproduction of free radicals, originating in molecular oxygen, may explain the lesions which result from inflammation, ischemia, and radiation exposure. Free radicals can cause damage to membranes, macromolecules, and DNA. Whether free radical production is mainly intracellular or extracellular may determine to a degree which kind of damage will occur. PMID- 6297067 TI - [Severe anaphylactoid complications in anesthesia. Apropos of 35 cases]. AB - 35 cases of severe anaphylactoid accidents during anesthesia are reported. Clinical symptomatology is dominated by cardiovascular collapse and/or bronchospasm and/or cutaneous and mucous manifestations. The course is always favourable. The usual risk factors (drug allergy, atopic background, spasmophilia, multiple anesthesia) are found. Pentothal alone or added to succinylcholine is most incriminated, followed by C-T 1341, curare-like agents, propanidid and macromolecular solutions. The immunoallergic investigation is incomplete, founded on the inhibition of leukocyte migration test TIML. The principal mechanisms responsible for these accidents and the most frequent causal products are recalled. Laboratory tests are reviewed. In conclusion, the authors propose a curative and preventive action to be taken when faced with this severe and non-exceptional accident of anesthesia. PMID- 6297068 TI - [Chemotherapy in advanced malignant melanoma. Results of a controlled trial comparing a combination of dacarbazine (DTIC) and detorubicin with dacarbazine alone]. AB - 51 patients with malignant melanomas who had not previously received chemotherapy were studied in a phase III trial. They were all advanced cases, either in relapse or showing metastatic spread and with one or several measurable tumor sites. They were judged to be too advanced for surgery or radiotherapy. They were split at random into two groups. Both groups received DTIC (250 mg/m2, IV, over 4 days every three weeks) and one group received detorubicin (120 mg/m2, IV every three weeks) as well. Detorubicin is a new anthracyclin drug which has shown promise in the treatment of these tumors in a previous trial. The combination of the two drugs produced better results than dacarbazine alone. Eight 50% regressions were obtained out of 22 cases studied (36%) with the combination, as opposed to 4 out of 26 (15%) with the single drug. The average length of response was also longer for the combination, i.e. 6 months opposed to 5 for the single drug. The differences were not, however, statistically significant (X2 = 2.09). Toxic reactions were also more frequent with the combination therapy (65%) than with DTIC alone (40%) (X2 = 0.42), as well as more serious because of the myocardial toxicity of the anthracyclin drug. This trial showed that a combination of dacarbazine with detorubicin improves the therapeutic outcome. Whether this represents a real benefit will need statistical confirmation from trials of combinations using other anthracyclins. PMID- 6297069 TI - [Significance of the presence of Australia antigen in liver diseases seen in a department of internal medicine in Abidjan]. AB - The exact responsibility of the hepatitis B virus in hepatic diseases seen in Ivory Coast is difficult to specify for several reasons, particularly technical ones. Nevertheless, we have tried to determine the prevalence of the HB virus in hospital practice and in patients hospitalized for liver diseases (viral hepatitis, cirrhosis, primary liver cancer). Comparison of the results to the prevalence of the virus in the ivorian population and statistical processing has led us to moderate the influence of HB virus in the different hepatic disorders. In primary liver cancer, the role of aflatoxine whose carcinogenic potential is now well established has not been disproved. PMID- 6297070 TI - [Hydatid cyst of the kidney. Apropos of 25 cases]. PMID- 6297071 TI - [Role of surgery in the treatment of thromboembolic disease and deep phlebitis of the legs. Apropos of 12 surgical cases]. AB - In reviewing a series of twelve cases treated surgically for deep venous thrombosis, with an admittedly short follow up (2 months to 2 years), the authors stress the beneficial effects of venous surgery. The postoperative mortality rate was zero and the morbidity rate marginal. They underline the importance of phlebographic exploration leading to earlier diagnosis, and list the therapeutic indications which vary according to the case and may include a supporting heparinotherapy and fibrinolytic treatment in a specialized center. They advocate venous surgery which has the twofold merit of preventing embolic complications and attenuating postphlebitic sequelae. PMID- 6297072 TI - [Blood biorheology and ischemic cerebrovascular accidents]. AB - Blood hyperviscosity is usually observed in ischemic stroke. Our purpose was to study relations between viscosity and the different parameters of blood that influence viscosity, and to search abnormalities of circadian rhythms. 18 stroke patients (8 females and 10 males mean 65.4 years) and 12 normal subjects (4 females and 8 males, mean 68 years) were studied. The measures were realised within the 48 hours after stroke at 6 p.m. and 12 p.m. and the fourth day at 6 p.m. and 12 p.m., by means of a Taylor-Couette type rheometer. Stroke patients have a significant increase of their blood viscosity, but without elevation of blood hematocrit or fibrinogen within 48 hours. Stroke patients have no nocturnal decrease of viscosity, and they don't present the normal circadian periodicity. Perhaps would this phenomenon explain the diurnal variations in occurrence of strokes? PMID- 6297073 TI - [Treatment of alcoholic patients with tiapride]. AB - The neuropsychiatric manifestations of alcoholism can be amended by neutralizing the somatic effects of alcohol on nervous centers. Tiapride acts electively on the mesolimbic area. Promising results have been obtained with tiapride in the various clinical forms of alcohol intoxication. Sixty patients (40 men and 20 women) were given tiapride for abnormal symptoms due to alcohol. Tolerance was good: no side-effects were recorded, with the exception of extrapyramidal manifestations in one patient. Symptoms due to alcohol abuse were alleviated. Relief of tremor was significant. Tiapride proved helpful in anxiety and depression and caused hallucinations to disappear in 35 cases. PMID- 6297074 TI - [Involvement of oculomotor nerves. A little-known complication of Horton's disease]. AB - Third, fourth and sixth cranial nerve palsy is a less common complication of giant cell arteritis than ischemic optic neuritis. Nevertheless, it occurs in at least 10% of cases. A case of giant cell arteritis with third nerve palsy which resolved under corticosteroid therapy is reported. The incidence of ocular complications is assessed from a review of the cases previously seen in the department and of data from the medical literature. The authors believe that the index patient has ischemic neuritis rather than myositis. It is concluded that giant cell arteritis should be considered at an early reversible stage. PMID- 6297075 TI - [Myoclonal encephalopathy due to dopamine]. AB - Onset of confusion with myoclonus occurred in a 55-year-old patient hospitalized for infectious endocarditis and under intravenous dopamine. The speculative responsibility of dopamine in the occurrence of this myoclonic encephalopathy is discussed. Indeed, although dopamine does not cross the blood-brain barrier, inflammation of the wall of the cerebral vessels (immunologic vasculitis) may have enabled dopamine to reach the central nervous system. PMID- 6297076 TI - [Immune deficiencies in nutritional anemias]. AB - A transient cellular immunologic defect caused by folic acid deficiency was seen in a goat-milk-fed infant with severe enterocolitis. Data on the immunologic consequences of folic acid, protein and iron deficiencies were reviewed in the medical literature. Investigations are difficult because of the patients' poor general condition. Results are difficult to interpret as many etiologic factors are often combined and mechanisms of immunologic responses are complex. Attention is drawn to the danger of iron therapy in patients with transferrin deficiency. PMID- 6297077 TI - [Distal bone metastases of the upper limb]. AB - This work is based on a review of 120 peripheral bone metastases of the upper limbs found in the literature and 10 others selected from a personal series of 469 cases of bone metastases. The authors stress the scarcity of these lesions (2%), metastases arising in bones of the forearm are usually part of a widespread osseous involvement, generally of mammary or prostatic origin. Metastases to the bones of the hand are more peculiar: 1) Those from bronchogenic carcinoma account for 50%. 2) Resultant soft tissue necrosis creates swelling, redness, heat and pain, which can mimic acute osteomyelitis. 3) X-rays reveal isolated osteolysis affecting metacarpals, phalanges, or more rarely carpal bones. Distal location of those metastases does not modify therapeutic management amputation is the best effective palliative treatment and must be discussed every time there is a concomitant skin ulceration, or if bone metastasis is unique or if the primary tumor is cured or accessible to treatment. PMID- 6297078 TI - [Value of electron microscopy in the diagnosis of viral infections]. PMID- 6297079 TI - [The H-Y antigen]. PMID- 6297080 TI - [Osteoporosis in pregnancy]. AB - The authors report two cases of osteoporosis, complicated by collapse of corpus vertebrae, which occurred during the last three months of pregnancy. Results of the study of phosphorus and calcium balance are not different from those found in other types of osteoporosis occurring in young adults. There is no antecedent osteopathy. Calciuria is normal. From analysis of previously published studies addressing the metabolism of phosphorus and calcium during pregnancy, a few pathogenic hypotheses for this rare aspect of osteoporosis are discussed. Special attention is given to calcitonin activity during late pregnancy. PMID- 6297081 TI - [Transhepatic fine needle cholangiography. Retrospective study of 60 patients]. AB - The purpose of this study was to present the retrospective results of 60 percutaneous transhepatic cholangiograms performed with a Chiba needle between 1978 and 1980 in a general radiology department. The puncture technique is now well established and the use of the Chiba needle considerably diminished the risk of local complications. The incidence of local complications is 5%. The complications are mainly due to the high pressure in the dilated bile ducts which can favorize infection and intraperitoneal bile effusion. When the right technique is used excellent imaging of the biliary system can be obtained. A correct diagnosis can be made in 95% of cases. The main indication of percutaneous transhepatic cholangiography is the obstruction of the main bile duct. The usual cases are: carcinoma of the head of the pancreas, carcinoma of the ampulla and other tumors of the main bile duct, neoplastic or inflamatory stenosis of the main bile duct, common duct stones and postoperatory complications of the biliary surgery. The errors of interpretation can be due to the difficulty to analyze the terminal portion of the bile duct or to false defects due to inhomogeneity of the contrast medium or a thick bile. The low complication rate and ist easy performance make the percutaneous transhepatic cholangiography competitive with the retrograde endoscopic opacification of the biliary system. PMID- 6297082 TI - [Parathyroid autotransplantation in hemodialysed chronic renal failure. Attempts to modulate secretion. First results apropos of 45 cases]. AB - The authors report their experience of parathyroid autotransplantation in 45 cases of renal insufficiency undergoing hemodialysis. The operation, carried out by two teams, reduces to the minimum the time during which the grafts are exposed to environmental factors likely to impair their function. An immediate anatomopathological examination is necessary to determine the degree of hyperplasia in each gland and the corresponding choice of grafts. The tissue to be implanted must be raised in the least hyperplastic gland and the size of the graft, always difficult to specify, is dictated by the intensity of the hyperplasia. This technique ultimately permits and almost permanent modulation of the parathyroid secretion by action on the secreting tissue. In the event of post operative persistent hypoparathyroidism, a second operation is desirable, using cryopreserved fragments for the implant. Conversely, in the event of clinical and biological recurrence, the ablation of a precise volume of grafted tissue can be performed without difficulty, under local anaesthetic. PMID- 6297083 TI - [Contribution of ultrasonics in the evaluation of abdominal tumors in children]. AB - Twenty-eight children with abdominal tumors were examined ultrasonically. Real time ultrasonography proved particularly useful in infants. In most cases ultrasonography shortened the diagnostic procedure by giving accurate images of the tumor's limits, size, contents, and origin. The method yielded an overall diagnostic accuracy rate of 89%. Moreover, ultrasonography is a reliable and safe means of monitoring abdominal tumors during and after therapy. In 1981, plain roentgenograms, intravenous urography with cavography and ultrasonography should be sufficient in the standard radiologic approach of most children with abdominal tumors. PMID- 6297084 TI - [The syndrome of compression of the left iliac vein at the promontory level; a new, serious anatomo-clinical entity?]. AB - A new case of pseudo-Cockett syndrome is reported. Both the clinical picture and the anatomic characteristics were unusual. Apparently, no similar cases have been reported in the medical literature. The relation between the cause of venous compression and it's results (thrombosis upstream and embolism downstream) could not be stated from data reviewed in the literature. No well-defined plan of management has been described. However, in the case reported here, therapeutic decisions gave satisfactory results. PMID- 6297085 TI - [Meningoencephalic and chorioretinal manifestations of toxoplasmosis in an immunodeficient patient]. AB - With reference to a case of malignant toxoplasmosis in a patient with angioimmunoblastic lymphadenopathy, the following features are recalled: in compromised hosts, clinical manifestations of malignant toxoplasmosis are highly variable, with neurologic signs and fever being predominant; the occurrence of lesions of the fundus, such as chorioretinitis, is an essential clue to the diagnosis; serologic tests for toxoplasmosis should be performed routinely at each stage of the disease in order to ascertain seroconversion through comparison with baseline results. PMID- 6297086 TI - [Systemic lupus erythematosus, polymyositis and toxoplasma antibodies]. AB - High toxoplasma antibody titers have been reported in polymyositis and other connectivitis with myositis. The authors describe a case of systemic lupus erythematosus with polymyositis and high titers of IgG toxoplasma antibodies. Studies of muscle biopsy specimens by immunofluorescence and inoculation of mice with muscle extracts failed to demonstrate the presence of Toxoplasma gondii. The nature of the toxoplasmic agression is discussed. It may be that reactivation of a previous infection due to the rupture of cysts was promoted by the immunologic disorders and complement deficiency related to systemic lupus erythematosus. PMID- 6297087 TI - [Treatment of early menopause. A report of 2 cases]. AB - Veralipride was given to two patients with early menopause ascertained by hormonal investigations. In both women (aged 33 and 30 years), an estradiol progestogen combination given for hot flushes, disorders of character, and depression, had been unsuccessful. Symptoms resolved under therapy with veralipride alone or associated with the previous treatment. In one patient, symptoms recurred after veralipride was discontinued and resolved again once the drug was resumed. PMID- 6297088 TI - [Clinical value of nephelometric assay of C-reactive protein]. AB - Laser immunonephelometry is a sensitive, specific and rapid method for assaying plasma C-reactive protein (CRP). This "acute phase" protein has recently been shown to be of significance in the regulation of immunological responses. It is released from hepatocytes early during infection, inflammation or necrosis. Nephelometry provides a good means for detection, monitoring and evaluation of therapeutic efficiency in these conditions. The increase in CRP may precede the positivity of the usual biological tests and is of value in situations where these tests cannot be interpreted. PMID- 6297089 TI - [Spinal meningeal hemorrhage of tumor origin]. AB - Intraspinal tumors with subarachnoid hemorrhage are exceptional. They are most often localized at the cauda equina. Ependymoma is the most common tumor. Low back pain followed by headache (Fincher syndrome) should suggest hemorrhage originating from a low lesion. PMID- 6297090 TI - [Study of erythrocyte bilirubin. Evaluation of the risk of kernicterus]. AB - 351 analysis of bilirubin binding by erythrocytes (BE) and of non conjugated bilirubin (NCB) are realised. 15 analysis have an increasing of the value: 6 with NCB and BE high with EST; 3 with NCB high and BE normal with EST; 6 with NCB high and BE normal without EST and any complication. PMID- 6297091 TI - [Study of thesaurismosis induced by perhexiline maleate. Confirmation of experimental data]. AB - Perhexiline maleate is an amphiphilic molecule. Along with many other drugs it is responsible for experimental and, in some instances, clinical lipidoses. Sphingomyelinase deficiency has been evidenced in cell cultures incubated with perhexiline maleate. We describe the occurrence of a similar defect in a patient. The disturbances in the phospholipid turnover which are responsible for the thesaurismosis may originate in the sphingomyelinase deficiency. PMID- 6297092 TI - [A new therapeutic approach to urogenital tuberculosis]. AB - The authors comment on the new treatment methods used in cases of urogenital tuberculosis. The progress made has led to a reduction in the duration of medical treatment. The introduction of rifampicine, and the combination of medical treatment with transurethral techniques, have to a great extent reduced surgery and consequently morbidity from urogenital tuberculosis. PMID- 6297093 TI - Quantitation of renal function with static imaging agents. PMID- 6297094 TI - The relationship between Epstein-Barr virus and lymphoma. PMID- 6297095 TI - Attitudes towards the rights of mental patients. A national survey in the United States. AB - Recent changes in mental health policy and treatment have largely been attributed to a professional concern for patients' rights. This view is challenged by patients' rights organizations and their allies, such as legal advocacy groups. These parties argue that psychiatric planners and providers are trying to coopt a popular movement and to use patients' rights as a convenient explanation for the mental health system's limited self-reform and fiscal limitations. A key issue here is the gap between planned policy and implemented practice. Patients' rights activists maintain that rights are only very sparsely implemented, while the mental health system believes that it has progressed quite far. A study of patients' rights attitudes held by the various involved forces can illuminate the conflicts in patients' rights policy and provide understanding of the potential for resolving these conflicts. Such an analysis also touches on some general themes concerning the relationships between health providers and social movements in the health field. This paper examines attitudes towards mental patients' rights on the part of mental patients rights groups, state departments of mental health, state hospitals and statewide mental health associations. One hypothesis was borne out: that patients' rights groups do not believe as much as does the mental health establishment that concern for patients' rights has played a large role in mental health policy. Further, the activist groups are more favorable than are the other respondents to abolition or sharp curtailment of intrusive psychiatric treatments. And, mental health officials and their lay supporters are less prone to accept patients' rights groups' initiative in reform efforts. The second main hypothesis, that the mental health establishment would be more antagonistic to patients' rights in states where patients rights groups existed, was not supported. One possible explanation for this is that patients' rights groups have had a nationwide impact, and that mental health officials face common problems throughout the country. Alternatively, or in conjunction, an antecedent variable of political liberalism may operate in some states, creating both activist groups and more liberal psychiatric professionals. The significance of these findings is discussed and suggestions are offered for future research in this area. PMID- 6297096 TI - [Adenovirus and para-influenza virus disorders]. PMID- 6297097 TI - Fatal systemic herpes simplex virus type 2 infection in a healthy young woman. AB - A previously healthy 28-year-old woman without historic or anatomic evidence of immunologic defect had a genital herpes simplex virus infection that rapidly progressed to systemic disease, with death 12 days after the onset of illness. Herpes simplex virus type 2 was isolated from the liver and spleen and was also shown by direct immunofluorescence in the liver and spleen, but not in the brain. We have found no previously reported instance of death from this virus in a young adult uncompromised immunologically. PMID- 6297098 TI - Synovial sarcoma of tonsil and tongue base. AB - We report a case of synovial sarcoma of the tonsil and tongue base. Radical surgery is the treatment of choice. Whether planned combination therapy may offer a better prognosis remains to be demonstrated. PMID- 6297099 TI - Influence of silica and carrageenan on spleen colonies and colonies in murine peritoneal cell-coated cellulose acetate membranes. AB - Cellulose acetate (CA) discs placed in the peritoneum of mice become coated by a layer of peritoneal cells consisting primarily of macrophages (M). These CAM support the growth of hematopoietic colonies when syngeneic bone marrow cells are injected intraperitoneally. Most of these colonies are granulocytic and are recognizable by their peroxidase reaction. Since silica and carrageenan are known to reduce macrophage function, their effect on CAM granulocyte colony formation was tested. Carrageenan injected intraperitoneally before, concurrently, or after injection of marrow cells markedly reduced colony formation. Silica injected intraperitoneally concurrently or after injection of marrow cells reduced colony formation. Silica injected before marrow cells did not reduce colony formation. CAM produced in one mouse and exposed to carrageenan or silica in situ for 24 h before being transferred to sublethally irradiated recipients and seeded by injection of marrow cells had control levels of granulocytic colonies. Likewise, CAM produced in one mouse, removed, incubated in vitro with carrageenan or silica, carefully rinsed and transferred to sublethally irradiated recipients and seeded with marrow cells were able to support control levels of colony formation. Intravenous injection of silica or carrageenan had no consistent effect on colony formation. Spleen colonies (CFU-S) from marrow cells incubated in vitro with the agents, and given intravenously to lethally irradiated mice, were inhibited by silica, but not by carrageenan. Silica or carrageenan given intravenously to irradiated mice 3 h before or 3 h after intravenous marrow cell injection enhanced subsequent CFU-S formation. PMID- 6297101 TI - [The significance of acral rewarming for the diagnosis of psychiatric diseases]. PMID- 6297100 TI - [Factors in the serum from patients with systemic lupus erythematosus enhancing superoxide generation by neutrophils]. PMID- 6297102 TI - [Changes in the hypothalamo-hypophyseal-adrenal (HHA) system in chronic alcohol abuse]. PMID- 6297104 TI - [Metabolism of purines and immune function]. PMID- 6297103 TI - [Therapeutic effects of glycine in tetanic syndrome (electromyographic and psychodiagnostic studies)]. PMID- 6297105 TI - Localization of insulinomas of the pancreas at operation by real-time ultrasound scanning. PMID- 6297106 TI - The significance of thrombocytosis in patients with carcinoma of the lung. PMID- 6297107 TI - Transplantation of rat insulinoma allografts with cyclosporin A. AB - Cyclosporin A (CyA), an investigational immunosuppressive drug, was tested for its ability as a single agent to prevent rejection and maintain function of islet cell tumors transplanted across a major histocompatibility barrier. Lewis rats have previously been shown by our group to reject the KX insulinoma in 7 to 10 days. CyA was administered to Lewis rats intraperitoneally for 21 days beginning 1 day before subcutaneous KX insulinoma engraftment. Dosage of 8 and 12 mg/kg/day failed to suppress rejection, as no palpable tumor or blood glucose level changes were observed. A dosage of 17 mg/kg/day allowed full allograft function without serious drug-related side effects in this short-term study. blood glucose levels in the successfully engrafted recipients fell to an average of 43 +/- 13 mg/dl. When CyA treatment was stopped, the insulinoma allografts were rejected within 14 days, suggesting that continued presence of the drug is necessary to maintain immunosuppression. These experimental results suggest that CyA can be administered as a single immunosuppressive agent to prevent early rejection of insulinoma transplanted across a major histocompatibility barrier in the experimental animal. PMID- 6297108 TI - Methods of repair of noncircumferential bile duct defects. AB - Although noncircumferential bile duct defects are uncommon, they are important because they require careful repair to avoid subsequent biliary stricture. I have encountered three of these defects in more than 1000 biliary operations. The method of repair chosen for a particular case depends on the pathologic defect, the potency of the ampulla, and the tissues available for use. If the ampulla must be bypassed, a Roux-en-Y jejunal reconstruction is applicable for most biliary defects. An anastomosis between the bile duct and duodenum may be suitable to repair low biliary defects. Occasionally, the gallbladder can be used as a conduit between the bile duct defect and the duodenum or jejunum. If the ampulla need not be bypassed, a Heineke-Mikulicz repair is suitable only for very short defects. A patch technique is a better choice for larger defects. Patches can be made of autogenous vein, gallbladder, knitted Teflon, or a serosal onlay patch of duodenum or jejunum. A vein patch is especially appealing because it will easily cover any extrahepatic defect. A different method was chosen in each of three cases. Side-to-side Roux-en-Y hepaticojejunostomy was used to repair a large cholecystocholedochal fistula with associated pancreatitis. Heineke Mikulicz repair was employed for a short hepatic duct stricture. Saphenous vein patch was used to repair a long bile duct defect during a left hepatectomy for hepatocellular carcinoma. This patient presumably represents the second successful reported vein patch repair and the only one with subsequent studies showing the fate of the vein patch. The vein patch apparently acts as a temporary scaffold allowing the outgrowth of biliary epithelium from the remaining bile duct wall. Careful initial repair of these noncircumferential bile duct defects is essential to avoid a subsequent biliary stricture with its disastrous consequences. PMID- 6297109 TI - Resolution of recurrent Verner-Morrison syndrome by resection of metastatic vipoma. AB - The case of a woman with recurrent Verner-Morrison syndrome associated with a multicentric and metachronously metastatic pancreatic islet cell tumor is described. Her original symptoms resolved after distal pancreatectomy and splenectomy for an islet cell carcinoma of the tail of the pancreas. Recurrent symptoms were associated with a raised plasma vasoactive intestinal polypeptide (VIP) level, an islet cell tumor in the neck of the pancreas, and liver metastases. After partial pancreatectomy and resection of the liver metastases, her symptoms resolved and plasma VIP level normalized. This case emphasized the potential role of resection in the management of the Verner-Morrison syndrome associated with a recurrent, metastatic vipoma. PMID- 6297110 TI - [Main achievements in the study of the pathogenesis and treatment of cholelithiasis]. PMID- 6297111 TI - [Small-cell cancer of the lung (clinical characteristics and the principles approaches to the choice of the therapeutic procedure)]. PMID- 6297112 TI - Perinatal death and respiratory apparatus dysgenesis due to a bis (dichloroacetyl) diamine. AB - N, N1-bis (dichloroacetyl) diamine 1, 8-octomethylenediamine (WIN 18,446) is an experimental drug which was first investigated as a male contraceptive. It is soluble in lipid solvents but not in water. The administration of 1,200 to 1,600 mg/kg to pregnant rats on the tenth day of gestation produced multiorgan fetal malformations. Smaller doses, 400 to 800 mg/kg, especially if divided over 2 or 3 days, caused perinatal death. Thus, 60 to 100% of offspring of rats given WIN 18,446 on the tenth and 11th days of gestation died at birth or within 4 days (Taleporos et al., 78). The present study investigated such deaths. At doses of 200 mg/kg on day 10 or 50 mg/kg on days 10 and 11, 67% of offspring had defective or absent diaphragms, 48% had tracheobronchiomegaly with cystic lungs, and 67% had pleural hemorrhage. At doses of 100 mg/kg given on 1 day or 25 mg/kg each day for 2 days, 50% had tracheobronchiomegaly with cystic lungs and rudimentary acini. At lower doses (18.8 mg/kg X 2 or 12.4 mg/kg X 3), a majority of fetal lungs had rudimentary acini, thick septa, few capillaries, and wide cuffs of perivascular connective tissue. Thus, a chemical given during organogenesis produced dysgenesis of the respiratory apparatus. Varying the dose produced malformed lungs with persistently deficient acini which model such human lung faults as tracheobronchiomegaly (Mournier-Kuhn Syndrome; Mounier-Kuhn, '32), bronchiolar dysplasia (Wilson-Mikity Syndrome), and perinatal death with acinar failure resembling neonatal hyaline membrane disease. PMID- 6297113 TI - [Surgery of the endocrine pancreas and endocrine gastrointestinal tumors]. PMID- 6297114 TI - [Diagnosis and therapy of endocrinically active ovarian tumors]. PMID- 6297115 TI - Small-cell carcinocythaemia. PMID- 6297116 TI - Fibreoptic bronchoscopy: effect of multiple bronchial biopsies on diagnostic yield in bronchial carcinoma. AB - The findings in bronchial biopsy specimens obtained at fibreoptic bronchoscopy in 271 patients with bronchial carcinoma were reviewed. Fifty-nine per cent of 703 specimens taken from the site of bronchoscopically visible tumours in 215 patients provided evidence of carcinoma. Unequivocal histological evidence of carcinoma was obtained in 78.6% of the 215 patients with visible tumours. When only one biopsy specimen was taken evidence of carcinoma was obtained in 65.2% of cases. At least five biopsy specimens were required to provide a greater than 90% probability of obtaining at least one positive sample. The anatomical site of the tumour had no significant effect on the proportion of biopsy specimens that were positive or the frequency of obtaining at least one positive sample. When extrinsic bronchial compression was the only visible abnormality evidence of carcinoma was obtained by bronchial biopsy in 26.8% of 56 cases. PMID- 6297117 TI - [Cytomegalus infections after heart surgery]. PMID- 6297118 TI - [Suspected non-palpable changes in the breast diagnosed by mammography. Radiological biopsy criteria and technic in marking the biopsy area]. PMID- 6297119 TI - [Suspected non-palpable changes in the breast diagnosed by mammography. Treatment planning and therapeutical problems]. PMID- 6297120 TI - [GABA and drug effects on the central nervous system]. PMID- 6297121 TI - [Direct and indirect lymphography in the dog]. PMID- 6297122 TI - Antiarrhythmic effects of disulfiram on epinephrine-induced cardiac arrhythmias in rabbits exposed to trichloroethylene. PMID- 6297123 TI - Basic phospholipase A2 from Naja nigricollis snake venom: phospholipid hydrolysis and effects on electrical and contractile activity of the rat heart. PMID- 6297124 TI - [Clinico-virological data in the diagnosis of recurrent herpetic stomatitis in children]. PMID- 6297125 TI - [Follow-up of patients with ovarial carcinomas from the point of view of the radiologist]. AB - During a radiotherapy of ovarial carcinomas and other gynecologic tumors, special attention must be given to the reaction of the organs of urinary excretion as well as of the small and large intestine, especially if the irradiation is applied by the moving strip method or as an abdominal bain with an irradiation field comprising the whole abdominal and diaphragmatic region. The early and late radiogenic reactions of our great number of patients were routinely examined. A continuous control of the urinary excretion conditions before, during and after radiotherapy is absolutely necessary. These examinations are also of high relevance with regard to the early recognition of recurrences. An additional control of the rectum and sigmoid region is necessary with respect to late lesions. Regular ultrasonic or CT examinations are important in order to early recognize metastases in the preferred regions of formation of metastases which are hardly accessible by other methods (intraperitoneal dissemination with ascites, surface of the diaphragm, local extension, retroperitoneal lymph nodes, and transdiaphragmatic extension to the interpleural lymph nodes). Finally the authors present their first experiences with the NMR technique in radio-oncology and discuss the value of tumor markers in case of ovarial carcinomas. PMID- 6297126 TI - Fast atom bombardment mass spectrometry of estrogen glucuronides and sulfates. AB - Fast atom bombardment (FAB) mass spectra of 13 intact, underivatized glucuronides and/or sulfate salts are reported. Spectra are characterized by abundant ions formed by attachment of a proton, [M+H]+, or of an alkali ion, [M+alkali]+, to the glucuronide or sulfate salt. Fragment ions were of low intensity. FAB spectra can be used to obtain the molecular weight of a sample, to assess its purity and to identify the nature of the alkali of the glucuronide or sulfate salt. PMID- 6297127 TI - Antagonism of the actions of estrogens, androgens and progesterone by anordrin (2 alpha, 17 alpha-diethynyl-A-nor-5 alpha-androstane-2 beta, 17 beta-diol dipropionate). AB - Anordrin, an antifertility agent that is an antiestrogen with weak estrogenic activity, has been studied to further characterize its hormonal activities. A dose of 2.0 micrograms/mouse X day for 7 days did not increase the uterine content of protein, but it did inhibit to a small extent the effect of administered estradiol-17 beta on uterine protein content and more significantly the effect of estradiol-17 beta on the uterine content of progesterone receptors. Anordrin also decreased serum corticosteroid-binding globulin levels. Administration of an average daily dose of 160 micrograms/day of anordrin to intact male mice had no effect on weights of kidney, testis, or seminal vesicle after 10 days, but seminal vesicle weight was significantly decreased after 30 days at a slightly lower dose. Similarly, anordrin inhibited the increase in seminal vesicle weight induced by testosterone propionate treatment of castrated mice. In female mice anordrin failed to maintain deciduomata and blocked the ability of progesterone (2.0 mg/mouse X day) to do so. However, anordrin did not compete with the androgen [3H]R1881 for binding in kidney cytosol or with the progestin [3H]R5020 for uterine receptor sites. Anordrin also did not compete with [3H]corticosterone for binding to serum proteins. PMID- 6297128 TI - The role of prostaglandins, cyclic nucleotides and tricarboxylic acids in the regulation of the human placental 20 alpha-hydroxysteroid dehydrogenase in vitro. AB - The in vitro effect of non-steroidal regulators (prostaglandins, cyclic nucleotides and tricarboxylic acids) on the cytoplasmic 20 alpha-hydroxysteroid dehydrogenase (20 alpha-HSDH, EC 1.1.1.149) isolated from human early gestational and term placentas was investigated. When prostaglandins (PG) were tested at 100 microM concentrations, an inhibition of the human placental 20 alpha-HSDH by PGE1 (80% inhibition), PGE2 (70%), PGF1 alpha (40%) PGF2 alpha (30%) and 13,14-dihydro 15-keto-PGF2 (PGFM) (20%) was observed. This effect was shown to be dose dependent. The I50 (concentration at 50% inhibition) was determined for PGE1 and PGE2 to be 11 microM and 38 microM, respectively. No effect on the activity of the 20 alpha-HSDH could be demonstrated for PGI2 and its stable metabolite 6-keto PGF1 alpha, for the cyclic nucleotides (dbcAMP, dbcGMP) and for the tricarboxylic acids (citrate, ketoglutarate, lactate, malonate, pyruvate and succinate) when added to the incubation at 100 microM concentration. The 20 alpha-HSDH isolated from early gestational and term placentas did not respond differently to the substances tested. These results suggest that prostaglandins can have a direct, dose-dependent effect on the isolated human placental 20 alpha-HSDH without cyclic nucleotides as intermediates and thereby play a role in the regulation of human progesterone synthesis and metabolism during pregnancy and near term. PMID- 6297129 TI - Identification of 5 alpha-stanols in patients with sitosterolemia and xanthomatosis: stereochemistry of the protonolysis of steroidal organoboranes. AB - Plasma and fecal sterols of patients who exhibited tendon xanthomas with normal plasma cholesterol levels were studied. Plasma levels were (mg/dl mean +/- SD): cholesterol 232 +/- 36; cholestanol 5.9 +/- 2.1 (normal less than 0.6); sitosterol 18 +/- 5.6 (normal less than 1.0); campesterol 11 +/- 3.3 (normal less than 1.0). Other sterols such as sitostanol and campestanol (the 5 alpha-dihydro derivatives of sitosterol and campesterol) were also present in large amounts. Examination of the feces from these subjects showed cholesterol, sitosterol, campesterol and their 5 beta-saturated derivatives. Presence of 5 alpha-stanols in the plasma, and their absence in the feces indicates that the 5 alpha-stanols are synthesized endogenously within the body rather than in the intestine by colonic bacteria. The absolute identification of the structures of these 5 alpha stanols was elucidated via hydroboration of their corresponding unsaturated sterols coupled with protonolysis of the resultant organoboranes. PMID- 6297130 TI - Mechanisms of immunological eradication of a syngeneic guinea pig tumor. II. Effect of methotrexate treatment and T cell depletion of the recipient on adoptive immunity. AB - The influence of methotrexate on the development of immunity to the line 10 hepatoma was studied in guinea pigs. Chronic methotrexate treatment had no apparent effect on the ability of immune guinea pigs to suppress the growth of inoculated tumor cells. In contrast, the same methotrexate regimen inhibited the development of tumor immunity if started before the 8th day after immunization with a vaccine containing viable line 10 cells admixed with Bacillus Calmette Guerin (BCG) cell walls. Thus, methotrexate selectively inhibited the afferent limb of the immune response. In adoptive transfer experiments, methotrexate treated recipient guinea pigs were capable of being passively sensitized with immune spleen cells, indicating that the primary cell-mediated immune response of the recipient was not required for adoptive immunity. The contribution of recipient T cells in adoptive immunity was further investigated in guinea pigs deleted of T cells by thymectomy, irradiation, and bone marrow reconstitution. Despite demonstrable deficiency in T lymphocyte reactions, "B" animals were fully capable of rejecting tumors after transfer of immune cells. These results suggest that the expression of adoptive immunity was independent of recipient T cell participation. In addition, sublethal irradiation of immune spleen cells prior to adoptive transfer abolished their efficacy. Proliferation of transferred immune cells in the recipient may be essential for expression of adoptive immunity. PMID- 6297131 TI - [Cyclic adenosine monophosphate and the enzyme activity of its metabolism in L cells sensitive and resistant to the cytotoxic action of ethidium bromide]. AB - Using L-cells both sensitive and resistant to cytotoxic action of ethidium bromide (EB), a study was made of the intracellular level of cAMP, activities of adenylcyclase, phosphodiesterase and cAMP, liberated from cells into the surrounding medium. In EB resistant L-cells compared to EB sensitive ones, the higher level of cAMP with a decreased activity of adenylcyclase and an increased activity of the phosphodiesterase was shown to be associated with an impeded exit of cAMP from cells. It is suggested that the differences in cAMP levels in the EB sensitive and resistant cells are associated with the properties of cAMP dependent protein kinases of these cells. PMID- 6297132 TI - [Localization of Na+,K+-ATPase activity in the cardiac myocytes of white rats at the ultrastructural level]. AB - The localization of Na+,K+-ATPase in cardiomyocytes of white rats at the ultrastructural level and the dependence of enzyme activeness on electronic density of structures, were studied. Na+,K+-ATPase activity was revealed in plasmalemma, Z-membranes and in nuclei. The enzyme activity is distributed discretely both within the tissue and within separate cells. It is, probably, due to the heterogeneity of structural-functional condition of cells and their ultrastructures as revealed by means of electronic microscopy in terms of their different electronic density. The maximal Na+,K+-ATPase activeness was revealed at the average electronic density of structures. PMID- 6297133 TI - Epidemiology of sheep pox. AB - An outbreak of sheep pox in a flock of 2,191 purebred indigenous sheep and their crosses with exotic European sheep had a course of 18 weeks. The peak incidence and mortality occurred 5 weeks after the onset of the epidemic. The morbidity was highest in the purebred and crossbred lambs and crossbred hoggets. The mortality rate and case fatality rate were highest in the crossbred hoggets. PMID- 6297134 TI - [Neurological complications in infectious mononucleosis]. PMID- 6297135 TI - [Paget's disease of the nipple. Histological variations]. PMID- 6297136 TI - Evaluation of parathyroid function in patients with hypergastrinaemia and pernicious anaemia. AB - In order to evaluate the possible causal relationship between raised serum gastrin levels and the development of primary hyperparathyroidism (HPT) which is suggested from experimental studies we evaluated parathyroid function in a group of 32 patients with hypergastrinaemia and pernicious anaemia. The values for serum calcium and parathyroid hormone were determined as well as the fasting urinary excretions of cyclic AMP and calcium. There was no relationship between the serum gastrin levels and any of the other studied parameters and there was no consistent pattern suggesting parathyroid hyperfunction. A retrospective analysis of hospital records from 441 patients operated for primary HPT showed a prevalence of pernicious anaemia of 1.8%. This figure is higher than that found in the unselected age-matched population (0.31%). However, taken together this study does not support the hypothesis that hypergastrinaemia is of particular importance for the pathogenesis of primary HPT. PMID- 6297137 TI - Effect of chemical modification of a histidine and a lysine residue of pea seed nucleoside diphosphate kinase. AB - Chemical modification of a histidine and lysine residue inactivates pea seed nucleoside diphosphate kinase (NDP kinase). Thus there seems to be a reactive lysine residue, at the active site of pea seed NDP kinase, in addition to the histidine residue phosphorylated by the substrate ATP as a consequence of the enzyme reaction. The presence of a reactive lysine at the active site of the enzyme could explain why a small amount of N-epsilon-phospholysine, as well as 1 phosphohistidine and 3-phosphohistidine, is formed on alkaline hydrolysis of the enzyme. PMID- 6297138 TI - Effects of chemical modification of lysine, tyrosine and tryptophan residues in pea seed nucleoside diphosphate kinase and inhibition of the enzyme with antibodies. AB - Pea seed nucleoside diphosphate kinase (NDP kinase) is an oligomeric, tetrameric enzyme which has been shown to be phosphorylated by its substrate ATP, presumably forming an 1-phosphohistidine at its active site. Recently has also a reactive lysine residue been demonstrated at the active site. In the present investigation nitration of a tyrosine residue is shown to inactivate the enzyme. There seems only to be a slight structural change in the enzyme on modification of these essential lysine and tyrosine residues, since double diffusion experiments with an inhibitory antiserum shows no difference in reactivity between the native enzyme and the modified enzyme. It is also found that modification of all tryptophan residues in the enzyme reduces the enzyme activity only to a small degree, indication that these hydrophobic amino acid residues are not directly involved in the catalytic process. PMID- 6297139 TI - CT scan in the diagnosis of urachal carcinoma. PMID- 6297140 TI - New treatment for urethral strictures. AB - A new operative technique using synthetic, absorbable mesh for grafting a urethral defect was applied in 7 mongrel male dogs. The ventral half of the urethral circumference with its surrounding corpus spongiosum was excised for a length of 3 to 4 cm. A Dexon mesh of the same dimensions, woven in our laboratory from polyglycolic acid fibers, was sutured to the defective area. A perineal urethrotomy was established, and no splints were left behind. Dogs were studied between two and six months. Retrograde urethrography showed that the operative area healed without strictures or irregularities. Intravenous urography showed no back pressure effects, and cultured urine was always sterile. Histologic examination two months after surgery showed that the urothelium was completely healed, without inflammatory changes or disruption. Suburothelial tissues were replaced by dense collagenous connective tissue. The excised corpus spongiosum did not regenerate. After six months, the area of dense collagen described was diminished in size so that the operative area could be hardly identified except by the absence of corpus spongiosum. PMID- 6297141 TI - Mucin-producing adenocarcinoma of bladder. Clinicopathologic report. PMID- 6297142 TI - Cultured rabbit vesical smooth muscle cells for lining of dissolvable synthetic prosthesis. AB - For prosthetic use in genitourinary operations we investigated the feasibility of producing grafts from cultured smooth muscle cells on absorbable synthetic biomaterial. Biopsies of bladder wall muscle without mucosa were obtained from 20 rabbits for culture of smooth muscle cells; cultured cells were examined by light and electron microscopy. Those cells were grown in subculture on a microfiber mesh carrier woven from 35-denier polyglycolic acid fibers. As shown by scanning electron microscopy (SEM), complete coverage of the synthetic fabric was achieved after ten to fourteen days of subculture. PMID- 6297143 TI - Malakoplakia of testis. AB - Two cases of malakoplakia of the testis are reported. Ultrastructural studies were performed in both cases and confirmed the histiocytic nature of the large cells seen in this granulomatous process. Characteristic Michaelis-Gutmann bodies are identified in those macrophages and seem to originate from altered lysosomes. From these observations and a review of the literature, the possible physiopathogenesis of the disease is summarized, and comments are made on the treatment of malakoplakia. PMID- 6297145 TI - [Cytological and cytochemical studies of cell cultures infected with the avirulent viral mutant MK 35 of the pseudorabies virus]. AB - Cytological and cytochemical studies were carried out of cell cultures of chick embryo fibroblasts infected with the vaccinal mutant strain MK 35 (4.8 X 10(7) PFU) of the Pseudorabies virus. It was found that the first cytologic changes in the infected cultures presented definite focal character. In the first hours of infection several rounded cells were included only, while later on an increasing number of cells were involved, and the foci grew in size. The cytoplasma of the infected cells contained inclusions which were small, spherical, basophilic, and Foelgen positive; six hours later the cytoplasmatic inclusions became larger, oval, acidophilic, surrounded by a brighter area, and were Folgen-negative. Parallel to the cytologic changes in the infected cell cultures there set in metabolic disturbances and changes in the enzymatic activity. The infected cells showed enhanced lactate dehydrogenase, diphosphopyridin-nucleotide diaphorase, thiamine pyrophosphatase, and alkaline phosphatase activity and suppressed succinate dehydrogenase and acid phosphatase activity. These studies were said to reveal new biologic properties of the vaccinal mutant virus. PMID- 6297144 TI - Detection of bovine leukosis virus in bronchoalveolar lung washings and nasal secretions. AB - Cattle and sheep persistently infected with bovine leukosis virus (BLV) were studied for the presence of the virus in bronchoalveolar lung washings and nasal secretions. The virus was demonstrated in the cellular fraction of the lung washings in six out of nine cattle and in one out of six sheep. In no instance was bovine leukosis isolated from the cell-free bronchoalveolar lung washings. The virus was isolated from the nasal secretion of only one of six naturally infected milking cows despite frequent sampling; the virus-infected nasal secretion was from a sick 10-year-old cow. Bovine leukosis virus was not isolated from cellular fractions of nasal secretions. PMID- 6297146 TI - [Immunoprophylaxis of parvovirosis in dogs (a review)]. PMID- 6297147 TI - Malignant fibrous histiocytoma arising in a recurrent chordoma. Case report and electron microscopic findings. AB - We present the case of a sacrococcygeal chordoma which recurred 15 years after the radical removal as a soft tissue tumor in the gluteal musculature. This tumor consisted of two parts: a chordoma without symptoms of aggressive cellular proliferation and a malignant fibrous histiocytoma. During the following 4 years several local recurrences of the malignant fibrous histiocytoma occurred in the gluteal musculature. The patient finally died of lung metastases. No chordoma tumor tissue was found in the lungs, in the gluteal musculature or in the sacrococcygeal bone area. Histology including electron microscopy revealed no proof of a transition of chordoma into malignant fibrous histiocytoma. It must be assumed that the secondary soft tissue tumor originated from residual chordoma cells which were implanted during the operation of the primary tumor. It remains unclear whether the malignant fibrous histiocytoma arose from mesenchymal stromal cells within the chordoma or directly from primitive neuroectodermal chorda cells which possess the ability to differentiate into a variety of cell types including mesenchymal cells. PMID- 6297148 TI - Retroregulation: control of integrase expression by the b2 region of bacteriophages lambda and 434. AB - Genetic fusions constructed by a combination of in vivo and in vitro techniques have been used to investigate the cis-regulatory role of a 250-base-pair segment of b2 DNA in the expression of int transcripts originating from three different promoters PI, PL, and PTRP. It has been established that (a) the PI and PTRP transcripts, which produce functional int protein, are efficiently terminated by the b2 segment, (b) in the same test system, the PL transcript, which does not result in functional integrase, is not terminated by the b2 segment, (c) this b2 (sib) effect does not depend on genes lying between int and N on the phage genome, (d) the sib effect is also observed with the b2 DNA of phage 434, which resembles that of lambda between -197 and att but diverges radically from it to the left of base -197. PMID- 6297149 TI - Intertypic recombination in poliovirus: genetic and biochemical studies. AB - Poliovirus strains of type 1 and type 3 carrying genetically mapped ts mutations and differring in growth response to guanidine have been used to infect HeLa cells. With four heterotypic pairs of the mutants, recombinants with the crossover points between the loci coding for the antigenic properties, on the one hand, and for the sensitivity to guanidine, on the other, have been obtained. The recombinants have been identified on the basis of their phenotypic properties and, in particular, of the pattern of inheritance of unselected markers. One recombinant has been characterized by fingerprinting virus-specific polypeptides. It has been found that the capsid proteins (VP2, VP3, and VP1) of this recombinant originate from the type 3 parent, whereas the nonstructural polypeptides (X, 2, and 4) are inherited from the type 1 parent. Implications of the poliovirus intertypic recombination are discussed. PMID- 6297150 TI - Altered pathogenicity of avian myelocytomatosis (MC29) viruses with mutations in the v-myc gene. AB - Avian myelocytomatosis virus MC29 inoculated into the wing web of 1-day-old Brown Leghorn chickens causes a high incidence of tumors, predominantly endotheliomas that are apparently induced directly by the action of the viral myc gene. Mutants of MC29, in which portions of the v-myc gene have been deleted and which have reduced ability to transform macrophages in vitro, induce few tumors, among which lymphomas and osteopetrosis predominate. Analysis of lymphomas from birds infected with mutant MC29 suggested that they were a result of helper virus action rather than the mutant MC29. PMID- 6297151 TI - The effect of dibutyryl cyclic AMP on restricted replication of rubella virus in rat glial cells in culture. AB - Normal adult rat glial (RG) cells in culture have been shown to restrict rubella virus (RV) replication, permitting the synthesis of only five of the seven intracellular viral polypeptides associated with a normal, productive infection. Recently uninfected RG cells have been reported to differentiate into mature glia in the presence of dibutyryl cyclic AMP (dB-cAMP), suggesting that the RG cells may be a continuous cell line of undifferentiated glioblasts. This communication describes the effect of dB-cAMP on rubella-infected RG cells. Following exposure to dB-cAMP, RV antigens could be demonstrated in the cytoplasm of the differentiating glioblasts, all seven intracellular RV polypeptides could be detected following immune precipitation, slab gel electrophoresis, and autoradiography, and infectious progeny virions could be titered from the tissue culture medium. These data indicate that dB-cAMP promotes glioblast maturation and a concomitant activation of RV replication. PMID- 6297152 TI - Isolation of a human pararotavirus. AB - During a survey of 350 different isolates of human rotaviruses, a single isolate with unique properties was obtained. Unlike all rotaviruses tested this unique isolate failed to react in an ELISA test with antibody directed against the group specific rotavirus antigen. The morphology of this unusual virus isolate was identical in electron micrographs with other rotaviruses and the virus also contained 11 double-stranded RNA segments characteristic of this virus group. The electrophoretic migration of these RNAs (electrophoretype) in polyacrylamide gels did not coincide with the typical pattern of distinct size classes observed with all human rotaviruses. The unique isolate thus had two distinct properties that distinguish it from human rotaviruses, lack of a group-specific antigen and an atypical electrophoretype of the RNA segments. Based upon this, the new human isolate is given the name pararotavirus to distinguish this new type of virus group. Porcine and chicken pararotaviruses have recently been described with similar properties. PMID- 6297153 TI - Transforming ability of avian defective leukemia viruses in early embryogenesis. AB - The response to infection of chicken hemopoietic cells derived from the early stages of embryogenesis by avian myeloblastosis virus (AMV) and avian erythroblastosis virus (AEV) was investigated. It was found that erythroid progenitor cells were present in the blastoderm at a higher frequency than that of myeloid progenitor cells. These results correlate with the observation that target cells for AEV were found to be more numerous than those for AMV. Therefore, blastoderm cells are of potential value in understanding the mechanisms of oncogenesis at the level of the target cells. PMID- 6297154 TI - The caprine arthritis-encephalitis virus is a distinct virus within the Lentivirus group. AB - The genetic relatedness among viral genomes of caprine arthritis-encephalitis virus, visna virus, and progressive pneumonia virus, was determined. Whereas the genomic RNAs of two strains of visna virus are indistinguishable as reflected both by their annealing kinetics as well as by the thermal stability of the hybrids, the caprine arthritis-encephalitis virus and visna virus have only 30% of their nucleic acid sequences in common. Furthermore, within the homologous regions of the two viral genomes, there is a significant level (approximately 10%) of mismatched base pairs. This limited homology that exists between caprine arthritis-encephalitis virus and visna virus was lower than the sequence homology observed between the genomes of visna virus and progressive pneumonia virus, or between the genomes of caprine arthritis-encephalitis virus and progressive pneumonia virus. All this indicates that caprine arthritis-encephalitis virus is an additional distinct member of the Lentivirus group of the Retroviridae family. PMID- 6297156 TI - Cloning of the terminal loop of vaccinia virus DNA. PMID- 6297155 TI - Similar transforming growth factors (TGFs) produced by cells transformed by different isolates of feline sarcoma virus. AB - Fisher rat embryo cells transformed by each of three independent isolates of feline sarcoma virus (FeSV) are shown to release transforming growth factors (TGFs) into cell culture medium. These acid- and heat-stable peptides compete for binding to, and stimulate phosphorylation of, EGF membrane receptors and promote anchorage-independent cell growth. Cells transformed by the Gardner and Synder Theilen strains of FeSV produce high titers of TGF (60-200 ng eq EGF/liter) while cells transformed by McDonough FeSV produce TGF at only low levels (less than 10 ng eq EGF/liter). Growth factors produced by cells transformed by each of the three FeSV isolates functionally and biochemically resemble each other, mouse sarcoma growth factor (SGF), and TGFs produced by human tumor cells. PMID- 6297157 TI - Heteroduplex analysis of cloned fragments of herpes simplex virus DNAs. PMID- 6297158 TI - Inhibition by vesicular stomatitis virus of herpes simplex virus-directed protein synthesis. AB - Infection of mammalian cells with either herpes simplex virus (HSV) or vesicular stomatitis virus (VSV) results in a marked inhibition of host protein synthesis. These viruses employ different mechanisms to turn off the host. In previous studies we showed that following infection with HSV, cellular mRNA was degraded and host polyribosomes were dissociated (Nishioka and Silverstein, Proc. Nat. Acad. Sci. USA 74, 2370-2374, 1977; Nishioka and Silverstein, J. Virol. 25, 422 426, 1978a). Degradation required synthesis of an HSV-specified polypeptide whereas dissociation appeared to be mediated by a heat-labile virion associated function (Nishioka and Silverstein, J. Virol. 27, 619-627, 1978b). In contrast, when cells are infected with VSV, host mRNAs are not degraded and polyribosome profiles are not drastically altered (Nishioka and Silverstein, 1978a). We have exploited the properties of these two viruses by infecting cells either simultaneously or sequentially in an effort to test our previous hypotheses. Analyses of the distribution of polyribosomes, stability of mRNA, synthesis of mRNA, and patterns of protein synthesis in coinfected cells permit us to conclude that dissociation of polyribosomes in cells infected with HSV results from expression of a virion associated function, degradation of cellular mRNA requires expression of the HSV genome, and VSV is dominant in doubly infected cells because it inhibits de novo transcription of the HSV genome. PMID- 6297159 TI - On the terminally redundant sequences of bacteriophage T3 DNA. AB - The nucleotide sequences of the termini of mature DNA from T3 phage were determined. A directly repeated sequence of 230 base pairs, corresponding to the terminal redundancy of T3 DNA, was found in the left and right end of the genome. The sequence of the terminal regions of T3 DNA was compared with that of T7 DNA. There are discrete sequences with significant homology, specifically in the regions around the two ends of the terminally redundant sequences. 5'-CCTAAAG and its variants appear frequently in both ends of the genome. T3 DNA fragments carrying the terminally redundant sequences were isolated from concatemeric DNA and analyzed by restriction enzymes. PMID- 6297160 TI - Differential reactivity of serum antibody from tumor-bearing 15I5 X 7(2) chickens for cross-reactive species of endogenous retroviral envelope glycoprotein. AB - Experiments were undertaken to determine if the sera of avian sarcoma virus infected 15I5 X 7(2) chickens exhibit antibody reactivity for species of endogenous retroviral envelope glycoprotein expressed in 15I5 X 7(2) fibroblasts. Two viruses were used for infection of the 15I5 X 7(2) chickens, Pr-A and cl. 85; the envelope glycoprotein of Pr-A, but not of cl. 85, is antigenically cross reactive with 15I5 X 7(2) endogenous retroviral envelope glycoprotein. Both the Pr-A and cl. 85-infected 15I5 X 7(2) chickens exhibited serum antibody reactivity for the envelope glycoprotein of the endogenous retrovirus RAV-O. In contrast, neither group of infected chickens exhibited detectable serum antibody reactivity for a distinct species of endogenous retroviral envelope glycoprotein, one which though antigenically cross-reactive with the envelope glycoprotein of RAV-O is expressed to much higher levels in 15I5 X 7(2) fibroblasts. Possible mechanisms to account for the observed pattern of antibody reactivity are discussed. PMID- 6297161 TI - Nongenetic complementation in VSV: asymmetric contribution of the L proteins of each parent in the rescue of group I ts mutants. AB - Complementation group I temperature-sensitive mutants of vesicular stomatitis virus (VSV) are rescued at nonpermissive temperature by UV-irradiated virus. Rescue is a nongenetic process mediated by the structural L protein molecules of the parental irradiated virus. This is shown by action spectra analysis (V. Deutsch, B. Muel, and G. Brun (1977), Virology 77, 294-305), efficiency of rescue at doses high enough to inactivate every gene I, and rescue by irradiated defective-interfering short particles. Viral molecular synthesis at 39.6 degrees using the ts genomes as templates and stimulated by UV-irradiated virus is shown by gel electrophoresis of rescued virion proteins. Parental strain ts 053(I) is thermolabile in vitro while the thermostability of the rescued virions, genetically characterized as ts group I virus, is identical to that of helper wt or ts+ revertant. This suggests that some parental Lwt molecules are reincorporated in the rescued virions, together with newly synthesized Lts molecules. Efficiency variation of the rescue as a function of the multiplicity of infection of the UV-irradiated virions depends on the m.o.i. of the unirradiated ts I mutants. This result suggests that rescue depends on the concentration of the helper L molecules and also of that of the L initially bound to the ts template. That L of both parents contribute to rescue is supported by the observations that (1) rescue by UV-wt is strongly diminished after in vitro heating of thermolabile ts O53(I). (2) Intragenic rescue can be demonstrated: The helper activity of a given UV-ts I mutant is different according to the unirradiated ts I mutant used; the activity of helper L associated with different templates in intragenic heterologous combinations is higher or lower than its activity in the homologous combination (self-rescue control), instead of being equal as expected. (3) Efficiency of rescue by UV-wt also varies according to the ts I mutant. The initially bound Lts seems therefore to play a role important and different from that of the helper L. Contribution of both parent L to rescue seems thus to be qualitatively different. PMID- 6297163 TI - Post-transcriptional control of human cytomegalovirus gene expression. AB - During the immediate-early, early, and late phases of human cytomegalovirus infection in human fibroblasts, transcripts accumulated, respectively, from approximately 20, 75, and 90% of the sequences on the genome. Not all of the sequences which accumulated during the immediate-early and early phases were represented on polysomes, however. Four transcripts synthesized in cycloheximide treated cells were studied in detail. A 2.2-kb transcript (0.713-0.733 map units) represented 95% of the polysome-associated RNA in cycloheximide-treated cells and was the first to be detected on polysomes at 2 hr postinfection in untreated cells. A second, less abundant, transcript of 5.2 kb (0.670-0.733 map units) was also found on polysomes in cycloheximide-treated cells, and preliminary evidence suggested that this transcript may be spliced during processing. A 3.25-kb transcript (0.190-0.217 map units) was identified also as a minor polysome associated species of RNA. One transcript of 4.8 kb (0.630-0.670) remained associated with the nucleus and was not processed into mRNA in cycloheximide treated cells. Differential stability between the various transcripts was observed, the 2.2-kb transcript being the most stable. The results showed that in human cytomegalovirus-infected cells controls exist at the level of transcript accumulation, transport into the cytoplasm, preferential association with polysomes, and relative stability of RNAs. PMID- 6297162 TI - Effects of the adenovirus H5ts125 and H5ts107 DNA binding proteins on DNA replication in vitro. AB - Genetic and biochemical studies of adenovirus (Ad) DNA synthesis in vitro demonstrate that the Ad DNA binding protein (Ad DBP) is not necessary for the initiation of Ad DNA synthesis but is required for chain elongation. The DBP, which enhances early elongation to the 26th deoxynucleotide by approximately two- to fourfold, is absolutely required as chain elongation proceeds further. Ad DNA synthesis was assayed in a system requiring Ad DNA covalently linked at each 5' terminus to a protein (Ad DNA-pro), various fractions of Ad-infected cytoplasm, and an extract of uninfected Hela nuclei. Initiation of Ad DNA replication was measured by the formation of a covalent complex between the 80,000 dalton preterminal protein (pTP) and 5' dCMP. DNA binding proteins from two ts mutants, H5ts125 and H5ts107, have been purified and shown to be functional at 30 degrees but inactive at 38 degrees in an in vitro elongation system dependent on purified proteins. Chymotryptic cleavage of the 72K wild-type Ad2 DBP produces a 34K carboxyl terminal fragment which retains full activity in the in vitro elongation of Ad DNA. PMID- 6297164 TI - Functions of the major nonstructural DNA binding protein of a herpesvirus (pseudorabies). AB - Eight mutants of pseudorabies virus belonging to complementation group 3 and situated between 0.14 and 0.18 units on the physical map of the genome were analyzed. All the mutants tested in this respect (seven) recombined with one another, indicating that the mutations were located in different regions of the gene. All mutants were DNA-; the first round, as well as subsequent rounds, of DNA replication was completely blocked at the nonpermissive temperature in the mutant-infected cells. After shift-up from the permissive to the nonpermissive temperature, viral DNA synthesis continued for a short period of time only and viral DNA which had accumulated at the permissive temperature became degraded. Parental viral DNA, however, retained its integrity at the nonpermissive temperature and viral DNA synthesis started immediately after shift-down of the mutant-infected cells from the nonpermissive to the permissive temperature (even in the absence of protein synthesis). All mutants belonging to complementation group 3 tested (5 out of 8) produced a thermolabile nonstructural DNA binding protein (136K). In some of the mutant virus-infected cells this protein failed to migrate to the nucleus. We conclude that the pseudorabies virus mutants in complementation group 3 code for a defective 136K protein and that this protein is not only essential to the process of viral DNA synthesis but also plays a role in the stabilization of progeny DNA (but not of nonreplicating parental DNA) within the infected cells. PMID- 6297165 TI - Structure and function of DNA binding proteins from revertants of adenovirus type 5 mutants with a temperature-sensitive DNA replication. AB - H5ts107 and H5ts125 are two adenoviruses type 5 (Ad5) mutants with a temperature sensitive DNA replication. Both mutants contain an altered gene encoding the DNA binding protein (DBP). We have established by nucleotide sequence analysis that both mutants carry exactly the same mutation in the DBP gene resulting in the substitution of a proline residue at position 413 in the wild-type DBP amino acid sequence (529 amino acid residues long) by a serine residue. Revertants of H5ts107 and H5ts125, which are temperature independent in plaque efficiency and growth in HeLa cells at 32 degrees and 39 degrees, were characterized by nucleotide sequence analysis of their DBP genes. Four types of revertants could be distinguished: revertants with the wild-type DBP amino acid sequence (type I) and, revertants carrying, in addition to the original H5ts107/H5ts125 mutation at position 413, intragenic second site mutations at position 508 histidine leads to tyrosine (type II), at position 352 glycine leads to aspartic acid (type III), and at position 347 alanine leads to proline (type IV), respectively. All intragenic second site mutations are located, together with the H5ts107/H5ts125 mutation, in the C-terminal 45-kD fragment of the adenovirus DBP molecule. This provides further evidence that this part of the DBP molecule plays an important role in viral DNA replication. Phenotypic characterization of the revertants (J.C. Nicolas, F. Suarez, A.J. Levine, and M. Girard (1981), Virology 108, 521 524; (J.C. Nicolas, D. Ingrand, P. Sarnow, and A.J. Levine (1982), Virology 122, 481-485) has shown that the second site mutations reveal additional functional domains in the DBP molecule. PMID- 6297166 TI - [Estimation of the prognosis of patients with ischemic heart disease with the aid of pyrophosphate scanning of the heart muscle]. PMID- 6297168 TI - [67Gallium citrate in the diagnosis of lung tumors]. PMID- 6297167 TI - [Effect of vitamin D3 and its metabolites 1,25-dihydroxycholecalciferol and 24,25 dihydroxycholecalciferol on callus mineralization in rats with femoral fracture]. AB - Additional administration of vitamin D3 at a physiological dose of 0.25 microgram daily into rats with femur fracture within 4 weeks did not affect the specific weight and chemical composition (content of Ca2+, P) in diaphyses of intact and impaired femurs as well as the content of Ca2+. Pi and activity of alkaline phosphatase in blood serum of the animals. At a higher dose 2.5 microgram daily vitamin D3 increased concentration of Pi in blood serum but did not alter the other parameters studied. Physiological doses of 1,25-dihydroxycholecalciferol (1.25 (OH)2D3) and 24,25-dihydroxycholecalciferol (24,25 (OH)2D3) (00.3 microgram and 0.25 microgram daily, respectively) did not affect the specific weight and composition of the impaired diaphyses, content of Ca2+ and activity of alkaline phosphatase in blood but increased slightly the Pi concentration. After a 5-fold increase in the dose of 1,25(OH)2D3 (0.15 microgram daily) specific weight and content of Ca2+ were decreased in the impaired bones with simultaneous increase in concentration of Ca2+, Pi and activity of alkaline phosphatase in blood serum. These data suggest that reparation was impaired under the conditions of acceleration of the bone tissue resorption. Increased doses of 24, 25(OH)2D3 (1.25 micrograms daily) stimulated the increase in specific weight and mineralization of the impaired bones and normalized the increased alkaline phosphatase activity in blood serum. Clinical examination of 24,25(OH)2D3 could be recommended as a drug stimulating the reparation under conditions of bone fracture. PMID- 6297169 TI - [Radioisotope study of liver function in diabetes mellitus]. PMID- 6297170 TI - [Cellular immunity study of multiple sclerosis in the lymphocyte blast transformation reaction to the viruses of acute human encephalomyelitis, herpes and measles]. AB - The blast transformation test revealed a high level of lymphocyte sensitization to human acute encephalomyelitis virus in 12 out of 29 (41.9%) patients with multiple sclerosis in the early stages of the disease and in the period of exacerbation in patients with long-term disease. The pattern of the blast transformation test in response to herpes simplex and measles viruses did not depend on the duration of the disease. High, moderate, and low levels of lymphocyte blast transformation reaction to herpes and measles viruses were observed in patients with multiple sclerosis with similar average durations of the disease. PMID- 6297171 TI - [Untypeable herpes simplex virus strains. DNA analysis]. PMID- 6297172 TI - [Sensitivity and specificity of fluorescent immunoglobulins for the herpes simplex virus]. PMID- 6297173 TI - [Hormones and tumor therapy: the rational basis of endocrine therapy in breast cancer]. AB - Since endocrine therapy has gained great importance in curative and palliative radiotherapy, the hormone-therapy of breast cancer owing to the relevance of the receptor contents in the tumour, has had to be reconsidered over the past decade. Accordingly, this paper deals with endocrine pathophysiology both in relation to the central and cellular mechanisms of hormone action; in addition it presents recent findings on biosynthesis of the different hormones. Furthermore, important treatment criteria with regard to patient selection will be listed. The discussion of these criteria is based both on clinical findings ("empirical criteria") and on the receptor status ("objective criteria") with a view to a better assessment of risk and prognosis at the time of surgery. PMID- 6297174 TI - [A 51-year-old asymptomatic female patient with a palpable tumor in the left upper abdomen]. PMID- 6297175 TI - Identification and determination of 8 alpha, 9 alpha-epoxyhexahydrocannabinol as a metabolite of delta 8-tetrahydrocannabinol of mice in vitro. AB - 1. 8 alpha, 9 alpha-Epoxyhexahydrocannabinol (8 alpha, 9 alpha-EHHC) was formed from delta 8-tetrahydrocannabinol (delta 8-THC) by mouse liver microsomal preparation in the presence of an NADPH-generating system. 2. The epoxide was identified by t.l.c., g.l.c. and g.l.c.-mass spectrometry and, together with 11 hydroxy-delta 8-THC (11-OH-delta 8-THC), was determined by g.l.c. 3. When delta 8 THC was incubated with mouse liver microsomal preparation, 8 alpha, 9 alpha-EHHC and 11-OH-delta 8-THC was formed to the extents of 14% and 23%, respectively, of the added delta 8-THC. PMID- 6297176 TI - [Morphologic reactions and long-term vasal tonus behavior of the coronary microvascular system in different states of spontaneous hypertension in the rat]. AB - The morphological reactions and the long-term vasotonus behaviour of the coronary microvasculature including its smallest ramifications as well as the relationships existing between these reactions and the myocardial activity in the different stages of spontaneous hypertension of rats (SHR) are unknown. Therefore 80 1-12 month-old male SH- and 60 normotensive Wistar control rats were histomorphometrically investigated. Already in the prestage of hypertension (1 month) an increase of wall thickness and phosphatase activity in blood vessels alpha less than 10,5 microns occurred. These appearances of vasotonus increase were regressed in the acute phase (2nd-4th month). When the rise of myocardial activity is lowered in the stabile stage the mentioned signs of vasotonus increase returned (6th month) and showed a strong spreading in the late stage (12th month), which is characterized by enzyme histochemically detected disturbances of heart muscle metabolism. The behaviour of long-term vasotonus of the microvasculature and myocardial metabolism in 6 month-old and older rats provides a hypothesis concerning the cause of the unknown increase of coronary contraction in the stabile phase. It points to the existence of a vicious circle which perhaps plays a role in the genesis of heart insufficiency in hypertension. PMID- 6297177 TI - [Metastasizing malignant histiocytoma]. AB - Report on a case of a 68-year-old woman suffering from a fast growing tumor, initially manifesting in the dermis. At first, the histological findings were judged as a spindle-cell sarcoma. Further histological investigations revealed that the tumor was a malignant histiocytoma. Tumors of this kind are rarely found in the scarce group of the sarcomas of the soft tissues. Since its identification as an entity, the interest in this malignant tumor has been increased. Other clinical, histological and electronmicroscopic findings in this patient are compared with those in the literature; the actual state of therapy is presented. PMID- 6297178 TI - [Polyfunctionality of the visual rhodopsin]. PMID- 6297179 TI - [Viremia in genital herpes]. PMID- 6297180 TI - [Warts of the feet and their treatment]. PMID- 6297181 TI - [Clinical observation of late syphilis with involvement of the heart and liver]. PMID- 6297182 TI - [Role of enkephalin-reactive system of the caudate nucleus in acquisition of alimentary conditioned reflex in dogs]. AB - Bilateral injections of 15 and 45 mcg of synthetic analogue of TYR-D-ALA-LEY Enkefaline-ARG into the body of the caudate nuclei in dogs produced a manifested increase in parameters of Pavlovian alimentary conditioned reflexes. Moreover, the dose of 15 mcg of LEY-enkefaline enhanced the background salivation and prolonged food consumption. Excited dogs became calm. During analogous injections into the right lateral hypothalamus the effect diminished while injections of 15 mcg of morphine hydrochloride were ineffective. The data point to role of the enkefaline-containing system in alimentary behaviour. PMID- 6297183 TI - Infectious complications of blood transfusion. PMID- 6297184 TI - Influence of fibre and Nebacitin on microbial activity and amino acid digestibility in the pig and rat. PMID- 6297185 TI - [Gel model for measuring crystallization inhibitor activity]. AB - In an attempt to develop a method with the most favourable possible relationship of information gained to expenditure of material and time a simple way of measuring crystallisation inhibitor activity was developed. Calcium and oxalate ions are diffused against each other out of two opposite start holes in a layer of gel on a microscopic slide and form a strip of calcium oxalate crystals where they meet in the middle. The substances influencing crystallisation are allowed to diffuse into this central area. The density and extent of the calcium oxalate strips vary according to the inhibitory effect of the substances used. These differences were registered by means of photometric measurement for comparison against a control without any inhibitor added and an inhibition index was computed. Some of the best-known inhibitors were tested and also show a defined effect in this model. In physiological concentrations magnesium and citrate can be said to have particular importance. The range of inhibitory activities to be expected of human urine is covered by the present model. PMID- 6297186 TI - [Hyperplasias and adenomas of the pituitary gland in an unselected autopsy material]. AB - 5,100 formalin-fixed pituitaries of an unselected autopsy material of the Institute of Pathology of the Ernst-Moritz-Arndt-University of Greifswald were analysed with regard to the occurrence of adenomas and adenomatous hyperplasias. Beside the histological classification the size was determined. For some adenomas of the pituitary gland (ACTH-cell-adenomas, TSH-cell-adenomas and evidently also prolactinomas) the development from a hyperplasia which is a consequence of a central regulation disorder is supposed. The pathogenesis of the other types of adenomas remains still unclear. Difficulties in differential diagnosis between adenomatous hyperplasias and small adenomas are discussed. PMID- 6297187 TI - Viral pollution of seawater at Barcelona. AB - A virological study of sea water was carried out at Barcelona (Spain) in June and September 1979. Three beaches were sampled in June; the September samples were taken in front of an outlet at 2, 5, 20 and 1,000 m offshore. All of the samples from the 3 beaches contained viruses, at rates varying from 0.12 to 1.72 MPNCU/l. Vaccinal poliovirus type 3 and coxsackie B5 predominated. For the offshore samples, 55% were positive with rates ranging from 0.15 to 0.36 MPNCU/l. Vaccinal poliovirus type 3 was clearly predominant. There was no significant difference in viral pollution rates between 2m and 1,000 m offshore. PMID- 6297188 TI - [Interaction of a cAMP-deficient Yersinia pseudotuberculosis mutant with a macrophage culture in vitro]. AB - The behavior of a virulent Y. pseudotuberculosis strain and its c-AMP-deficient mutant possessing lower virulence in their in vitro interaction with the culture of peritoneal macrophages of white mice, as well as a change in the level of c AMP in macrophages in the process of the phagocytosis of the initial and mutant strains were studied. The c-AMP-deficient mutant was shown to retain its ability to infect macrophages and to multiply in them, while loosing its cytotoxic effect. Macrophages obtained from mice previously immunized with the mutant acquired the capacity of resisting the cytotoxic action of the initial virulent strain. The mutant proved to be unable to elevate the level of c-AMP in macrophages in the process of phagocytosis, while the initial strain caused a considerable increase in the level of c-AMP in macrophages. PMID- 6297189 TI - [Method of obtaining an erythrocyte diagnosticum for detecting Cl. perfringens type A enterotoxin]. AB - Erythrocytic immunoglobulin diagnostic reagent for detecting Cl. perfringens enterotoxin in the indirect hemagglutination test has been obtained. The diagnostic reagent has been prepared on the basis of antibodies of Cl. perfringens enterotoxin with the use of acroleinized tanned sheep red blood cells. The indirect hemagglutination test with this reagent is characterized by high specificity and sensitivity (the method permits to detect enterotoxin protein at a concentration of up to 0.005 microgram/ml). The preparation thus obtained is stable and ensures highly reproducible results. PMID- 6297190 TI - [Patterns in the spread and serovirological diagnosis of serous meningitis]. AB - The epidemiological and clinico-etiological study of cases of acute serous meningitis with unknown etiology in children was carried out in a large industrial city at the period of a considerable morbidity rise caused by this infection. The maximum morbidity was registered among younger children under school age attending children's institutions. In 26 closed groups of children group morbidity was revealed (4 cases and more), in 5 such groups small local outbreaks were registered. The clinico-instrumental methods of study permitted one to differentiate the groups of children having serous meningitis of supposedly enteroviral etiology, and sero virological studies carried out with the use of a wide range of diagnostic reagents revealed the etiological role of group B Coxsackie virus, mainly type 4, in 20.6% of cases, ECHO virus, serotypes 3 and 11, in 20.7% of cases, and parotitis virus in 5.3% of cases. PMID- 6297191 TI - [Protective properties of theta-hemolysin obtained by affinity chromatography]. AB - The data on the study of the protective activity of theta hemolysin and Cl. perfringens lecithinase preparations and the corresponding antitoxic sera obtained by indirect immune affinity chromatography are presented. Experiments in mice and guinea pigs indicate that the injection of antihemolytic serum and immunization with anatheta hemolysin ensures the protection of the animals from theta toxin. The enrichment of analecithinase preparation with anatheta hemolysin has been found to increase its protective properties against Cl. perfringens culture and toxin. PMID- 6297192 TI - Endogenous inhibitor of a cyclic AMP-independent (A type) protein kinase. AB - The cytosol fraction of the cells of Saccharomyces cerevisiae contains a low molecular-mass, heat-stable inhibitor for endogenous cAMP-independent protein kinase. The inhibitor (Mr 15 000) is specific toward the protein kinase of A type, while the protein kinase of G type and the catalytic subunit of cAMP dependent protein kinase are not affected. The following results suggest a protein structure of the inhibitor: 1. preincubation of the inhibitor with trypsin totally abolished its activity; 2. the inhibitor can be labelled by reductive alkylation of the amino acids with [14C]formaldehyde and sodium cyanoborohydride. The kinetic experiments have shown that the inhibitor is a competitive effector of the protein kinase of A type with respect to the protein substrate. PMID- 6297193 TI - Effects of phospholipase C on gastric vesicle membranes containing H+, K+-ATPase. PMID- 6297194 TI - Study of Epstein Barr virus (EBV) antibodies: IgG and IgM anti-VCA, IgG anti-EA and Ig anti-EBNA obtained with an original microtiter technique: --serological criterions of primary and recurrent EBV infections and follow-up of infectious mononucleosis--seroepidemiology of EBV in Belgium based on 5178 sera from patients. PMID- 6297195 TI - [Hepatoma with ectopic ACTH secretion]. PMID- 6297196 TI - Fine needle aspiration biopsy of deep cerebrum. A comparison of normal and neoplastic morphology. AB - Fine needle aspirates of deep white matter, basal ganglia and thalamus from necropsies of three neurologically normal men were compared with two aspirates of cerebral infarcts and with eight aspirates from neoplasms that occurred in the deep cerebrum. Normal white matter contained oligodendroglia with small, dark, round nuclei and astrocytes with larger, bland, oval nuclei and occasional cytoplasmic extensions. Basal ganglia and thalamus contained, in addition, numerous neurons with large nuclei and prominent nucleoli. Recognition of these normal components of the deep cerebral structures is necessary to avoid erroneous diagnoses of neoplasia. PMID- 6297197 TI - Metastatic breast carcinoma in cerebrospinal fluid. A cytomorphometric study. AB - A study was undertaken to quantitate the cellular characteristics of metastatic breast carcinoma in cerebrospinal fluid (CSF). Millipore filters of CSF from 15 patients with metastatic breast carcinoma were reviewed; 50 cells per case were evaluated when available. All cells in all cases shed singly or in loose clusters; tight balls or morulae were absent. All cells had regular, round-to oval nuclei with finely granular chromatin. The majority of cells in all cases had single or multiple round nucleoli, granular cytoplasm with distinct borders and a mean nuclear-cytoplasmic ratio of close to 0.70. Cellular background, number of tumor cells per case, number and placement of nuclei and nuclear and cytoplasmic diameter varied both within and among the cases. There was significant variation in nuclear and cytoplasmic diameters both within and among the cases of infiltrating ductal carcinoma. Thus, the uniform appearance of the cells was due to consistent cytologic features, not to similarity in cell size. The cytologic profile of metastatic breast carcinoma is sufficiently characteristic to distinguish this tumor from other benign and malignant lesions that shed in the CSF. PMID- 6297198 TI - Cytomorphology of the variants of small-cell carcinoma of the lung. AB - Cytologic classification of subtypes of small-cell carcinoma of the lung (SCCL) was retrospectively performed on respiratory material of 90 cases of lung carcinoma cytologically diagnosed as SCCL. Additionally, the cytologic material was reexamined for the presence of variants of SCCL in 68 cases of lung carcinoma whose types had not been conclusively defined by cytology. The type of lung cytology specimens reviewed were sputa, bronchial washings and brushings and fine needle aspirations. The study included review of pretreatment tumor histology, when present, and examination of the ultrastructure of the tumor in selected cases whose type had not been well defined by histology. Subtype recognition depended on the adequacy of the specimens rather than on the type of cytologic material examined. The cytologic subtyping was generally in agreement with the histologic subtyping of the tumor, except for cases in which SCCL was combined with other types of lung carcinoma, where certain discrepancies were noted. PMID- 6297199 TI - Diagnostic cytology seminar. PMID- 6297200 TI - Ultrastructural characterization of human corticotrophs (ACTH cells). AB - Ultrastructural characterization of human corticotrophs (ACTH cells) was performed by 'superimposition technique', which enabled detailed ultrastructural observation of immunoreactive ACTH cells in adjacent semi-thin light microscopic immunoperoxidase and routine electron microscopic sections. The human corticotrophs were large and round or polygonal and were not stellate. They had scanty rough endoplasmic membranes and were packed with numerous large secretory granules measuring from 250 to 500 nm in diameter. The sizes of secretory granules in 6 human pituitaries were 448 +/- 128, 344 +/- 86, 448 +/- 117, 244 +/ 65, 316 +/- 76, and 340 +/- 93 nm, respectively. The granules were not seen in a single row along the plasma membrane as is the case in the rat. They possessed somewhat irregular outlines with a rarely discernible halo. Different densities of granule matrices were occasionally found. The cells often contained a few large heterogeneous vacuoles. From these findings, the human ACTH cells were recognized to be remarkably different in cell shape and size, properties of secretory granules and cytoplamic inclusions from those of the rat pituitary gland. In respect to secretory granule properties, the human ACTH cells are similar to those of some other mammals (fox, young pig, and lerot). More data is required to elucidate the relationship between human ACTH cell morphology and functional state. PMID- 6297201 TI - Synthesis and storage of hormonal granules in cultured cells of the rat pituitary pars intermedia. AB - Tissue from the pars intermedia of the pituitary from new-born and adult rats was maintained in tissue culture for periods up to 14 days. Both new-born and adult rat pars intermedia cells showed active production of secretory granules during tissue culture, and also showed formaldehyde-chloral induced fluorescence. Immunohistochemically, beta-endorphin could be demonstrated in cultured pars intermedia cells. In contrast to the appearance of the pars intermedia cells in vivo, the granules in the cultured cells were most often seen along the cell membrane, and the cytoplasm was often dominated by clear vacuoles slightly larger than the granules. No exocytotic figures were observed in the cultured cells. The mechanism of hormonal release from the organ, if release does occur, thus remains obscure. In conclusion, cultured cells from the rat pituitary pars intermedia continue their production of hormonal granules in tissue culture, and thus form a valuable tool for further investigations on the complex regulation of hormonal secretion form the organ. PMID- 6297202 TI - Gonadotrophin-secretion in adrenocortical insufficiency: impact of glucocorticoid substitution. AB - In patients with deficient endogenous glucocorticoid production due to primary adrenal insufficiency (n = 4) or bilateral adrenalectomy (n = 2) a rise in LRH stimulated concentrations of LH was seen following withdrawal of substitution therapy for 84 h. Consecutive re-administration of glucocorticoids (dexamethasone 2.0 mg/day for 3 days) resulted in increased basal concentrations of LH and FSH and a diminished secretory response of LH upon iv LRH (100 micrograms). Five patients substituted with glucocorticoids because of adrenocortical insufficiency presented upon the administration of exogenous ACTH1-24 with unchanged basal and LRH-stimulated concentrations of LH and FSH as compared to a placebo experiment. These data suggest that the withdrawal and subsequent re-administration of gluco corticoid substitution alters basal and stimulated concentrations of gonadotrophins in patients with adrenocortical insufficiency, thus providing evidence for the importance of adequate glucocorticoid supply in the regulation of gonadotrophin secretion. PMID- 6297203 TI - Modification by Su-10'603 of the response to histamine on the production of cortisol and corticosterone in isolated canine adrenocortical cells. AB - The effect of Su-10'603 on the cortisol and corticosterone response to histamine in dispersed canine adrenocortical cells was examined. The production of cortisol and corticosterone in isolated dog adrenocortical cell incubations increased in a dose-dependent manner at concentrations of histamine between 0.01 and 10.0 micrograms/ml medium, with the predominant production of cortisol. When Su-10'603 (0.005 to 5.0 micrograms/ml medium), an inhibitor of 17 alpha-hydroxylation on corticosteroidogenesis, was added in cell incubations with histamine (1.0 micrograms/ml medium), the production of cortisol decreased in a reciprocal fashion, with a concomitant increase of corticosterone production. At the concentration between 0.05 and 0.1 microgram Su-10'603/ml medium, there was a shift from cortisol to corticosterone as the predominant product. These results indicate that in the dog, histamine at pathophysiological concentration stimulates steroidogenesis and acts at a relatively early stage in the biosynthetic pathway. PMID- 6297204 TI - Effect of gonadotrophin-releasing hormone (GnRH) and GnRH agonists upon accumulation of progesterone, cAMP and prostaglandin in isolated preovulatory rat follicles. AB - To study the acute and direct effects of GnRH agonists preovulatory follicles were isolated from PMSG-treated immature rats and incubated for 15-360 min in modified Kreb's bicarbonate buffer. The levels of cAMP, prostaglandin E, and progesterone were analysed in the tissue and/or incubation media. GnRH and two GnRH agonists produced a dose-dependent stimulation of progesterone production with maximal levels 5-6-fold higher than the control group. As compared to LH the magnitude of this effect was small and was detected only after 240-360 min of incubation. GnRH also stimulated prostaglandin E accumulation and this effect was as pronounced as for LH. There were no detectable changes in cAMP levels for any concentration of GnRH when the incubation time varied between 15 and 120 min whether or not a phosphodiesterase inhibitor was present, but after 240 min of incubation a 2-fold increase in cAMP was found. Consistent with previous results, LH caused a pronounced (40-50-fold) increase in follicular cAMP which was already detectable after 15 min of incubation. Indomethacin abolished the rise in prostaglandin E induced either by GnRH or LH but did not affect the response in terms of cAMP or progesterone, and did not affect the stimulation of meiotic maturation of the follicle-enclosed oocytes caused by the hormones. It is concluded that GnRH can exert acute and LH-like stimulatory effects on the preovulatory rat follicle but that the mechanism of GnRH action is different from that of LH. PMID- 6297205 TI - Comparison between the progestin secretion responsiveness to gonadotrophins of rat cumulus and mural granulosa cells in vitro. AB - Several studies have shown differences in gonadotrophin receptor content between different granulosa cell (gc) populations within the ovarian follicle, but little is known about the gonadotrophin sensitivity in gc from different parts of the follicle. This study is an investigation of progestin synthesis and responsiveness to highly purified human and partly purified rat gonadotrophins. It involves short-term culture of rat cumulus and mural gc obtained from preovulatory follicles of PMSG-treated immature rats. The responsiveness to FSH in terms of progestin accumulation was similar in the two gc populations, whereas the responsiveness to LH was greater in the mural gc than in the cumulus. The morphological response in the cumulus cells (mucification) correlated with the steroidogenic response. The pattern of progestin synthesis differed between the two gc populations. In the cumulus gc progesterone was dominant, whereas 20 alpha dihydroprogesterone was the main progestin secreted in the mural gc. The difference in responsiveness to gonadotrophins correlates well with the reported differences in receptor content in the two gc populations. PMID- 6297206 TI - Relationship of follicle stimulating hormone (FSH)-sensitive adenylate cyclase activity to FSH binding in immature rat ovaries following administration of pregnant mare serum gonadotrophin. AB - The time and dose-response relationships of human follicle stimulating hormone (hFSH)-sensitive adenylate cyclase activity to hFSH binding was studied in the immature rat ovary following an sc injection of pregnant mare serum gonadotrophin (PMSG). When an optimal dose of PMSG (10 IU/rat) was administered, a marked increase in hFSH-sensitive activity was observed at day 2, followed by a sharp decline at day 3. This was accompanied by a parallel rise and fall in ovarian hFSH binding activity. When immature rats were given various doses (5-100 IU/rat) of PMSG for 2 days, hFSH-sensitive adenylate cyclase activity increased sharply and maximal stimulation was obtained at 10 IU/rat. A close correlation was also observed with respect to dose-response for hCG-sensitive adenylate cyclase and hCG binding activities. It is concluded that: 1) PMSG administration with an optimal dose to the immature rat induced ovarian FSH and LH-hCG receptors, and an adenylate cyclase system highly sensitive to hFSH and hCG, and 2) the acquisition and responsiveness of adenylate cyclase to gonadotrophins are closely related to the appearance and the numbers of gonadotrophin receptors. PMID- 6297207 TI - Effects of various replacement oestrogens on hepatic transcortin synthesis in climacteric women. AB - The influence of peri-menopausal oestrogen replacement therapy upon protein synthesis in the liver was investigated. Changes in the cortisol-binding capacity of transcortin (TC-BC) were utilized as an indicator of hepatic oestrogen sensitivity. Forty-two climacteric women were studied before and after treatment with various drugs at different doses (patients receiving identical compounds served as their own controls for the evaluation of dosage effects; duration of medication: 14 days at each dose). Oral administration of oestriol (1 and 2 mg daily; n = 6), oestriol (1 and 2 mg daily) + oestradiol-17 beta (2 and 4 mg daily; n = 8), oestradiol valerate (1, 2, 4 and 6 mg daily; n = 6), conjugated equine oestrogens (0.3 and 0.6 mg daily; n = 6), sodium oestrone sulphate (0.8 and 1.6 mg daily) + sodium equilin sulphate (0.2 and 0.4 mg daily; n = 6) and quinoestrol (25 micrograms daily; n = 6) failed to alter TC-BC. However, ingestion of higher doses of conjugated equine oestrogens (0.9 and 1.25 mg daily; n = 5) and quinoestrol (50 micrograms daily; n = 6) caused a significant rise (P less than 0.01). Ethinyloestradiol was given orally as a reference substance and elicited an elevation of expected magnitude. Vaginal administration of dienoestrol cream (1 mg daily; n = 4) did not change TC-BC. These data indicate that the hepatotrophic effects of replacement oestrogens vary depending on the preparation and dosage selected for treatment. PMID- 6297208 TI - Trilostane in the management of Cushing's syndrome. AB - Trilostane has been used to treat 6 patients with Cushing's syndrome, 4 with an adrenal adenoma and 2 with pituitary driven disease. Three patients responded clinically and biochemically to trilostane. Although effective in some cases of Cushing's syndrome, the variability of the effect of trilostane may limit its usefulness as a therapeutic agent. PMID- 6297209 TI - Inhibitory effect of cyproheptadine on ACTH secretion in patients with Addison's disease. AB - In 14 patients with Addison's disease plasma levels of ACTH were studied after administration of a single oral dose (16 mg) of cyproheptadine. The drug administration was followed by an inhibition of ACTH release. These results support the view that cyproheptadine may exert an inhibitory effect on ACTH secretion in subjects whose corticosteroid feedback mechanism is normal. We hypothesize that the effect of cyproheptadine might be related to its anti serotonin or anti-histaminergic action. PMID- 6297210 TI - Is the adrenal angiotensin receptor angiotensin II--or angiotensin III like? AB - In order to determine whether the adrenal receptor is primarily directed at angiotensin II (AII) or angiotensin III (AIII) the following in vitro experiments were performed examining aldosterone responsiveness in isolated glomerulosa cells. 1) Cells exposed to increasing doses (2.4 X 10(-10)M - 2.4 X 10(-6)M) of AII or AIII were found to be significantly more responsive to AII (AII's ED50 was 6.3 X 10(-10) M vs AIII's 4.6 X 10(-9)M P less than 0.001). 2) Octapeptide analogues (Sar1 Ala8 and Asn1 Ala8), while demonstrating different inhibitory potencies relative to each other, were equally effective in blocking AII vs AIII stimulation. 3) The heptapeptide analogues (des1 Ala8 and des1 Ile8) however, inhibited AIII stimulation preferentially (P less than 0.01). 4) The 8 alanine octapeptide analogues were better inhibitors of both AII and AIII stimulation than the 8 alanine heptapeptide analogue. 5) HPLC analysis indicated that AII and AIII were being degraded at the same rate during the incubation procedure. These results, taken together, strongly suggest that the angiotensin adrenal receptor is an AII receptor. PMID- 6297211 TI - Changes in the responsiveness of adrenal cells from foetal goats at different times during pregnancy. AB - To understand the factors responsible for the increased pre-partum output of cortisol from the adrenal glands of the goat foetus, we measured changes in responsiveness to ACTH in vitro of dispersed adrenal cells from foetal goats of various gestational ages and we determined the output of cortisol (F) and corticosterone (B) in the presence of exogenous progesterone (P4) and 17 alpha hydroxyprogesterone (17 alpha-OHP4). The increment in F output after ACTH increased 5-fold between day 100 and days 147 or 154 (term). This increase was not associated with changes in the ED50 for ACTH stimulation of F output. The basal and ACTH-stimulated output of F was similar to that of B at day 100 but 5-7 times greater than that of B at day 154. There was a significant effect of ACTH on P4 output on days 77, 100 and 147 but net utilization of P4 by the cells at term. The ratio of exogenous P4: 17 alpha-OHP4 converted to F varied with gestation and increased from 0.11 at day 100 to 0.99 at day 154. Exogenous P4 was converted to B at all stages of pregnancy. We conclude that in the goat there is an increase in the responsiveness of foetal adrenal cells to ACTH between day 100 of gestation and term. One factor associated with this change in adrenal responsiveness may be an increase in the activity of the enzyme 17 alpha hydroxylase. PMID- 6297212 TI - Effects of angiotensin II on the zona glomerulosa of sodium-loaded dexamethasone treated rats administered or not with maintenance doses of ACTH: stereology and plasma hormone concentrations. AB - The trophic effects of angiotensin II on the rat zona glomerulosa were investigated by stereological and radioimmunological methods. Angiotensin II was not able to reverse the sodium-loading and dexamethasone-induced inhibition of the growth and steroidogenic capacity of the zona glomerulosa. However, if the sodium-loaded dexamethasone-treated rats were given maintenance doses of ACTH, angiotensin II exerted a strong adrenoglomerulotrophic effect. The hypothesis is advanced that the co-operation of ACTH and angiotensin II is required in the maintenance of the morphologic and enzymatic integrity of the rat zona glomerulosa. PMID- 6297213 TI - Pharmacological effects of 2- and 4-methyloestradiol as a probe to test the biological importance of 2- and 4-hydroxylation of oestrogens (catecholoestrogen formation). AB - 2-Methyloestradiol, 4-hydroxy-2-methyloestradiol and 4-methyloestradiol were synthesized and infused sc into long-term ovariectomized rats by means of osmotic minipumps. The effects on uterine growth and gonadotrophin release were tested and compared to oestradiol or vehicle alone. 2-Methyloestradiol and 4-hydroxy-2 methyloestradiol, steroids in which the 2-hydroxylation is selectively blocked, definitely increased uterine weight (dry and wet), decreased morning LH and increased evening LH serum levels. 4-Methyloestradiol, a compound which cannot be 4-hydroxylated in vivo, was inactive in the model used. It is assumed that 4 hydroxylation constitutes an essential step in the expression of oestrogenicity of primary oestrogens, whereas 2-hydroxylation does not. PMID- 6297214 TI - A quantitative assay for the cytoplasmic androgen receptor using [3H]dihydrotestosterone in the presence of NAD+-nucleosidase. AB - In prostatic cytosol DHT1 is metabolized to 5 alpha-androstane-3 alpha (or beta), 17 alpha-diols with a half life of 2 h even at 4 degrees C. Thus, [3H]DHT appears to be a poor marker for a quantitative assessment of androgen receptors (AR). Methyltrienolone (R1881) seems to be advantageous as it is not metabolized. However, because of considerable binding to progestin receptors, assays using [3H]R1881 are not specific for AR in tissues containing progestin receptors. We, therefore, developed a specific assay for AR using [3H]DHT (14 nM) as marker, where metabolism of DHT is prevented by pre-incubation with NAD+-nucleosidase. The [3H]DHT-receptor complex is separated from free, SHBG-bound and unspecifically bound [3H]DHT by agar gel electrophoresis. The binding sites of high affinity and low capacity are characterized by suppression with unlabelled R1881 (2 microM) in a parallel assay. Under these conditions DHT and R1881 appear to have the same kinetics of association and dissociation. Weighted non-linear regression analysis of specific binding capacity at various ligand concentrations reveals that in rat prostatic cytosol the affinity of DHT (Kd = 0.405 +/- 0.0839 nM) is significantly higher (P less than 0.01) than that of R1881 (Kd = 1.25 +/- 0.271 nM). PMID- 6297215 TI - beta-Endorphin-like peptides in autopsy pituitaries from adults, neonates and foetuses. PMID- 6297216 TI - Anti-thyrotrophin receptor antibodies in Graves' disease as demonstrated directly by immunoprecipitation assay. AB - An immunoprecipitation assay was developed to determine the presence of antibodies against human TSH receptors. With this assay we were able to demonstrate that in comparison with sera from normal controls. 24 out of 30 (80%) sera from patients with untreated Graves' disease could immunoprecipitate more [125I]TSH-TSH receptor complexes. In 9 assays, an average of 14.1 +/- 3.7% (SD) of the [125I]TSH-TSH receptor complexes was immunoprecipitated by the 30 Graves' sera vs 9.8 +/- 3.0% by the normal pool serum (n = 23) (P less than 0.001) and 7.7 +/- 2.8% by the 22 normal sera (P less than 0.001). One serum of the 24 positive Graves' sera was studied in detail. The results suggest that this serum contained an anti-TSH receptor auto-antibody directed towards a different determinant on the TSH receptor than the TSH binding site. PMID- 6297217 TI - Somatostatinoma syndrome: does a clinical entity exist? AB - We report here 2 patients with somatostatin-secreting tumours and hypersomatostatinaemia. One subject, a 36 year old woman with diabetes, flushing, labile blood pressure and diarrhea, had elevated basal plasma levels of somatostatin-like immunoreactivity (SLIR) and calcitonin. Plasma SLIR increased further following tolbutamide administration. Plasma levels of prostaglandin E2 (PGE2) and pancreatic polypeptide (PP), normal in the basal state, showed exaggerated responses to pentagastrin and secretin, respectively. Immunocytochemistry of the tumour tissue revealed cells containing somatostatin-, calcitonin-, PGE2- and PP-like immunoreactivity. The other patient, a 52 year old male, had an SLIR-secreting tumour of the proximal duodenum and elevated basal and post-tolbutamide SLIR levels but no signs or symptoms suggestive of increased SLIR production. Tumour tissue revealed cells containing somatostatin- and calcitonin-like immunoreactivity. We conclude that patients with somatostatinomas do not always exhibit a predictable syndrome. Patients with these tumours may exhibit a range of clinical, biochemical and immunocytochemical features typical of endocrine tumours of mixed-cell origin, such that the dominant signs and symptoms associated with these neoplasms cannot readily be ascribed to overproduction of any single hormone. PMID- 6297218 TI - Studies on the release mechanism for hypothalamic hormones. PMID- 6297219 TI - [Acute infectious diarrhea]. PMID- 6297220 TI - Coexistence of alpha-thalassemia and a new pyruvate kinase variant: PK Fukien. AB - A 12-year-old male of Chinese ancestry had life-long hemolytic anemia attributed to alpha-thalassemia. Restriction endonuclease mapping of his DNA revealed that in reality, he had three alpha-globin loci, but he was homozygous for pyruvate kinase deficiency. The new pyruvate kinase variant carried by this patient was characterized and designated PK Fukien. PMID- 6297221 TI - Retinal projection to the inferior colliculus in the rat. PMID- 6297222 TI - Meningiomas with a non-meningotheliomatous component. A new type of tumour? AB - Seven cases of meningiomas with "pseudopsammoma bodies" have previously been described in the literature. Two additional cases are presented. Electron microscopy of the cells surrounding the "pseudopsammoma bodies" reveals an ultrastructure different from that of the meningotheliomatous cells. It is concluded that meningotheliomatous meningiomas with "pseudopsammoma bodies" are mixed tumours, including a non-meningotheliomatous component, the origin and significance of which is uncertain. PMID- 6297223 TI - Ependymitis, leukoencephalitis, hydrocephalus, and thrombotic vasculitis following chronic infection by mouse hepatitis virus 3 (MHV 3). AB - Mouse hepatitis virus 3 (MHV 3) is either avirulent (resistant mice), hepatotropic (susceptible mice), or neurotropic (semisusceptible mice), depending on the strain of mice infected. In semisusceptible mice, infection led first to a transient meningitis, ependymitis, and leukoencephalitis, followed by a permanent communicating hydrocephalus and, later on, to a chronic thrombotic vasculitis affecting meningeal and parenchymal vessels at the brain stem level. Small foci of ischemic necrosis related to vascular occlusions were seen in the dorsal brain stem. Cyclophosphamide treatment of semisusceptible mice significantly reduced the meningeal infiltrates but did not prevent the development of hydrocephalus and other neuropathologic changes. Identical lesions occurred in fully susceptible mice infected with a low dose of virus, but no neurologic disorder could be induced in genetically resistant mice even following immunosuppression or intracranial inoculation. The leukoencephalitis differed from the demyelinating lesions observed with MHV 4. Vascular lesions were of particular interest. More attention should be given to the possibility of virus induced chronic cerebral vasculitis in man. PMID- 6297224 TI - Globoid cell leukodystrophy: specialized contact of globoid cell with astrocyte in the brain of twitcher mouse. AB - Subplasmalemmal linear densities (Yajima et al. 1977 a) were the membrane specializations observed in globoid cells in globoid cell leukodystrophy (GLD) and in the cells of the mononuclear phagocytic system (Kawanami et al. 1980). In the spinal cord of the twitcher mouse, an authentic murine model of GLD, somewhat similar membrane specializations were noted in astrocytes, and on some occasions, a spot desmosome-like cellular contact was observed between globoid cells, which were likely to be mesodermal in origin, and astrocytes, which are of ectodermal origin. Possible significance of such apparent cellular contact is discussed briefly. PMID- 6297225 TI - Morphological features of Pick's and atypical Alzheimer's disease in Down's syndrome. AB - This is a pathological analysis of a 50-year-old severely mentally retarded female with trisomy 21 who five years prior to her demise developed progressive dementia, epileptic seizures and choreiform movements. The necropsy revealed gross and microscopic features of Alzheimer's and Pick's disease. Ultrastructurally the majority of neurofibrillary changes and all studied Pick bodies consisted of 15 nm straight tubules. A few neurofibrillary changes were composed of 22 nm paired helical filaments, which were also found in the enlarged neurites of neuritic plaques. A few paired helical filaments were interspersed between straight tubules of Pick bodies. These findings in a patient with Down's syndrome and previous reports of atypical features of Alzheimer's disease indicate that very probably our case is a variant of Alzheimer's disease, thus broadening the spectrum of pathological changes observed in Alzheimer's disease. PMID- 6297226 TI - Computerized classification of gliomas by automated microscope picture analysis (AMPA). PMID- 6297227 TI - Histological observations on the regrowth of malignant gliomas after radiotherapy and chemotherapy. AB - The reproliferation of glioblastomas after radiation and chemotherapy has been studied in 25 tumors by means of whole mount histological preparations. The tumors have been subdivided into four groups according to the radiation dose and the distance from the end of radiation. After 4,000 rad vessel proliferations and mitoses stop, while vessel degenerations and astrocytes with deformed nuclei appear. Six months after 6,000 rad have been delivered, repopulation phenomena are clearly evident and are mainly represented by mitoses both in parenchyma and in the vessel walls, circumscribed necroses with pseudopalisadings, proliferations of formerly degenerated vessels, etc. The brain adjacent to tumor (BAT) has a great importance since it is one of the reproliferating sites, even though it may be unrecognizable for the occurrence of radiation damages. PMID- 6297228 TI - An immunocytochemical study of intraneuronal inclusions of the caudate and substantia nigra. Reaction with an anti-actin antiserum. AB - The composition of the eosinophilic inclusions of the large neurons in the human caudate nucleus and of neurons in the substantia nigra was investigated by immunocytochemical methods. Sections of caudate nucleus and substantia nigra were stained using a peroxidase-anti-peroxidase method with antisera to actin, to neurofilament proteins, and to a crude CNS microtubule fraction, the last of which reacts with neurofibrillary tangles. Of the several antisera, only the anti actin antiserum gave positive results, indicating that these inclusions, composed of regularly arranged filaments, are highly ordered aggregates of actin polymers. PMID- 6297229 TI - Hirano bodies in myelinated fibers of hepatic encephalopathy. AB - Inclusions, essentially identical to Hirano bodies, were observed in the substantia nigra, dentate nuclei, and frontal lobes in three autopsy cases of hepatic encephalopathy. Similar inclusions were not observed in these areas in nine controls. The inclusions were mainly seen in the inner loops of the myelinated fibers, between the myelin lamellae, and among degenerated myelin sheaths. The same inclusions were rarely observed in postsynaptic terminals. PMID- 6297230 TI - Hilus cells from human postmenopausal ovaries: gonadotrophin sensitivity, steroid and cyclic AMP production. AB - Strips of ovarian hilar tissue taken from 11 post-menopausal women were examined histologically and found to contain clusters of hilus cells in contiguity with non-myelinated nerve fibers. No other steroidogenically active structures were found. Specimens from these strips were incubated for 30 and 240 minutes in Krebs bicarbonate buffer containing 5.5 mM glucose and 1% bovine serum albumin. The specimens produced measureable amounts of androstenedione, estradiol-17 beta and progesterone in vitro. The major steroid formed was androstenedione, and this pattern of steroidogenesis appeared similar to that of postmenopausal ovarian stroma. However, the amounts of steroids formed were considerably higher than in stromal tissue. This implies that the hilus cells possibly may be of greater importance, qualitatively, than the stroma cells for steroidogenesis in the postmenopausal ovary. Addition of hCG to the incubated specimens elicited increase in both cyclic AMP formation and steroid synthesis, especially of estradiol-17 beta, indicating a preserved responsiveness to gonadotrophin in hilus cells from postmenopasual ovaries. PMID- 6297231 TI - Spectral sensitivity measurements performed with the Goldmann perimeter. PMID- 6297232 TI - Ultrastructure of intracytoplasmic inclusion bodies in hepatocytes of griseofulvin-treated mice. Fibrils with myelin figure and rod-like fibrils. AB - Intracytoplasmic eosinophilic inclusion bodies light-microscopically identical to human alcoholic hyalin were produced in hepatocytes of griseofulvin-treated mice. Electron-microscopic investigation on these inclusion bodies revealed two morphologically different fibrils. The most frequently observed fibrils in liver tissues appeared straight or slightly curved in longitudinal sections and granular in cross sections. High-power electron micrographs on these fibrils revealed that each fibril consisted of parallel-running fine filaments of about 1.25 nm thick which were separated by spaces of about 1.25 nm. The ultrastructural appearance of these fibrils resembled that of myelin sheath in longitudinal sections. They usually measured from 13 to 32 nm wide and up to 0.9 micron long. Fibrils with larger diameter of up to 62 nm were infrequently encountered. Rod-like fibrils ultrastructurally similar to those seen in human alcoholic hyalin were also found in isolated materials. All these fibrils were isolated in the same fraction by a slight modification of a discontinuous sucrose gradient ultracentrifugation and a two phase polymer centrifugation. The present observation suggests that intracytoplasmic inclusion bodies produced in this animal model are composed of morphologically different fibrils. These different fibrils could share the same origin. PMID- 6297233 TI - Isolated adrenocorticotropic hormone (ACTH) deficiency. AB - The case to be reported is that of a 72-year-old woman with isolated adrenocorticotropic hormone (ACTH) deficiency, who complained of anorexia and generalized malaise. The secretions of human growth hormone (HGH), prolactin (PRL), luteinizing hormone (LH), follicle stimulating hormone (FSH), and thyroid stimulating hormone (TSH) were all within normal limit. In spite of the extremely low level of cortisol, the plasma level of ACTH would not rise sufficiently, but a marked response of cortisol to ACTH stimulation was recognized. The postmortem examination revealed a decrease in basophilic or PAS-positive cells of the anterior pituitary gland which also showed a selective loss of ACTH-secreting cells over immunohistochemical study. Electron microscope could easily visualize somatotroph, mammotroph, thyrotroph, FSH- and LH-gonadtroph, but corticotroph was difficult to be discerned. Adrenocortical cells demonstrated atrophy and degeneration, for which the zona fasciculata and zona reticularis were narrowed. The zona glomerulosa was slightly enlarged in width. PMID- 6297234 TI - Hepatocellular carcinoma combined with hepatic mesenchymal cell tumor and sarcoma. AB - An autopsy case of hepatocellular carcinoma combined with sarcoma and mesenchymal cell tumor or cholangiole cell carcinoma is presented. The main tumor nodes in the liver consisted of hepatocellular carcinoma and sarcoma. Extrahepatic metastases showed characteristics of monocytic or reticulum cells and hemangioendothelial sarcomas. In this report, some problems concerning hepatocellular carcinoma and sarcomatous representation of the liver tumor are proposed. Differentiation to the monocyte-macrophage-like cells and a reticulum cell pattern were present in the metastatic locations. Thus strongly suggesting that the sarcoma may have arisen from mesenchymal cells, particularly from the Kupffer cells or endothelia consisted of the sinusoid. However, mesenchymal tumor from cholangiole cells might be associated in a part of the sarcomatous cells. PMID- 6297235 TI - Liposarcoma. A clinicopathologic subtyping of 52 cases. AB - Fifty-two cases of liposarcoma were analyzed clinicopathologically, and were grouped into the following five types: myxoid (28 cases), well-differentiated (14 cases), pleomorphic (4 cases), round-cell (3 cases), and mixed (3 cases). Three of the 28 myxoid tumors showed a pleomorphic pattern resembling that of the myxoid variant of malignant fibrous histiocytoma, and one of the 3 round-cell tumors was considered to be a malignant counterpart of hibernoma. Ages ranged from 22 to 86 years with the average age of 49 years, the average patients with myxoid or round-cell liposarcoma being over 10 years younger than those with well differentiated or pleomorphic liposarcoma. There was also a variation in the site of predilection of the cases: the great majority of myxoid type tumors occurred in the thigh, popliteal fossa and buttock (23 cases), while the majority of well differentiated type tumors were seen in the retroperitoneum (10 cases). The rate of local recurrence of the tumor was 46%. Four of the 5 well-differentiated liposarcomas which recurred exhibited dedifferentiated pleomorphic components resembling those in malignant fibrous histiocytoma. The prognosis was more favorable in patients with ordinary myxoid tumors than in those with well differentiated tumors. The overall relative five-year survival rate was 63%. PMID- 6297236 TI - Immunohistochemical observation of intracytoplasmic lysozyme in proliferative and neoplastic fibrohistiocytic lesions. AB - Distribution of intracytoplasmic lysozyme in proliferative and neoplastic fibrohistiocytic lesions and non fibrohistiocytic tumors was studied by immunoperoxidase technique on formalin fixed, paraffin-embedded sections. The cases examined were 161 fibrohistiocytic lesions and 86 non-fibrohistiocytic tumors. Contrary to our expectation, the lysozyme positive cells were found only in the minority of cases with fibrohistiocytic lesions. Cells positive for lysozyme were found only in 13 out of 100 cases of dermatofibroma, one out of 4 cases of xanthogranuloma and 8 out of 33 cases of malignant fibrous histiocytoma. Dermatofibrosarcoma protuberans and non-fibrohistiocytic tumors were negative for lysozyme. It is suggested that in proliferative fibrohistiocytic lesions, induction of lysozyme synthesis is weak or absent. Some malignant fibrous histiocytomas showed scattered lysozyme positive neoplastic cells, indicating their probable histiocytic origin or differentiation. On the other hand, evidence of histiocytic differentiation of dermatofibrosarcoma protuberans was not obtained using lysozyme immunohistiochemistry. PMID- 6297237 TI - [Preparation of [3H]-labelled N-[1-(beta-hydroxy beta-phenylethyl)-3-methyl-4 piperidyl]-N-phenylpropionamide and studies of its binding with opiate receptors of mouse brain]. PMID- 6297238 TI - [Progress in receptor pharmacology in China]. PMID- 6297239 TI - Hormones, receptors, cyclic nucleotides and calcium. Presentations at the Seventh Scandinavian Meeting on Cyclic Nucleotide Research. Sundvollen, September 15.-17. 1982. PMID- 6297240 TI - The role of adrenergic receptors in Ni2+-induced coronary vasoconstriction. AB - The possible involvement of adrenergic receptors in nickel ion (Ni2+)-induced coronary vasoconstriction was studied on isolated perfused rat hearts and on isolated canine coronary artery strips. The experiments on both models showed that (i) alfa-adrenergic blockade by phenoxybenzamine or phentholamine caused only partial depression of Ni2+-induced coronary vasoconstriction: (ii) beta adrenergic receptor blockade by propranolol totally prevented Ni2+-action, and (iii) Ni2+ (1 microM) caused significant inhibition of coronary vasodilatation induced by isoproterenol. The experimental results indicate that alfa adrenoceptors play minor role (if any) in the coronary action mechanism of Ni2+ but it may be mediated by beta-adrenergic mechanisms. Nickel was found to alter the reactivity of coronary beta-adrenoceptors suggesting a possible modulatory role of this trace metal in coronary adrenergic mechanisms. PMID- 6297241 TI - Dependence of nickel-induced coronary vasoconstriction on the activity of the electrogenic Na+, K+-pump. AB - The possible interactions between the vasoactive trace metal nickel ion (Ni2+) and membrane Na-K-ATPase in the isolated perfused rat heart and in the isolated canine coronary artery have been studied. The characteristic features of 1 microM Ni2+-induced contractile response in the canine coronary artery strip were similar to those evoked by the inhibition of Na-K-ATPase. Inhibition of the pump activity by ouabain (10(-4)M) or by K+-deficient Krebs solution prevented Ni2+ action both in the canine coronary artery strip and in the perfused rat heart, indicating that when Ni2+ causes coronary vasoconstriction the Na, K-exchange is influenced. Further studies are needed to clarify whether Ni2+ acts directly on the enzyme, or the vascular action of this trace metal depends on the ionic gradients maintained by the electrogenic Na-K-pump. PMID- 6297242 TI - Effect of induced phase transitions on the glucose-6-phosphatase activity and electron transport of rat liver microsomes. PMID- 6297243 TI - Leukotriene B4 induces extravasation and migration of polymorphonuclear leukocytes in vivo. PMID- 6297244 TI - Leukotriene C4-induced cyclic nucleotide changes and contractile response in guinea pig trachea. PMID- 6297245 TI - Myocardial accumulation of 99Tcm-pyrophosphate and 99Tcm-gluconate compared with morphologic findings in daunorubicin treated rabbits. AB - In long-term daunorubicin treated rabbits increased myocardial accumulation of 99Tcm-pyrophosphate and 99Tcm-gluconate of a varying degree were recorded, visible at gamma camera examination in more than half of the animals. Chronic cardiomyopathy morphologically and topographically different from the ischemic myocardial injury was demonstrated in most animals at light microscopic examination. The myocardial abnormalities were classified in qualitative and quantitative scores and compared with the degree of isotope accumulation. The rabbits receiving a large single dose of daunorubicin had slightly increased isotope accumulation in the myocardium but no histopathologic changes. PMID- 6297246 TI - Herniography. The diagnosis of hernia in the groin and incompetence of the pouch of Douglas and pelvic floor. PMID- 6297247 TI - Morphologic classification of non-Hodgkin's lymphoma. I. Retrospective analysis using the Kiel classification. AB - In a retrospective analysis of 334 cases of non-Hodgkin's lymphoma observed between 1969 and 1978, 250 cases could be classified according to the Kiel classification. Clinicopathologic correlation was analysed for these latter cases. Irrespective of the morphologic appearance, all cases initially in stage I showed an excellent prognosis after radiation therapy alone, whereas the prognosis for stage II was similar to stages III and IV. For stages II-IV, 3 major prognostic groups with significantly differing survival curves were identified. The median survival times were one, 3 and more than 7 years, respectively. The pathologic and clinical significance of the Kiel classification is discussed. PMID- 6297248 TI - Primary hypothyroidism as a complication after treatment of tumours of the head and neck. AB - In 100 successive patients treated for tumour in the head and neck region, the function of the thyroid gland was evaluated during follow-up by determination of the serum levels of thyroid stimulating hormone and thyroxine. Radiation therapy alone did not lead to hypothyroidism, 4 patients had no dysfunction after laryngectomy for recurrent tumour after irradiation. Ten of 17 (59%) patients treated with irradiation and surgery including hemithyroidectomy and with a follow-up period of more than one year developed functional disturbance; in the absence of hemithyroidectomy one of 10. In patients who underwent hemithyroidectomy and developed hypothyroidism, the interval between surgery and postoperative irradiation was shorter than in those who did not develop functional disturbance (31 versus 49 days). PMID- 6297250 TI - Carcinoma of the uterine cervix. Incidence and mortality in the Stockholm-Gotland region. AB - During the years 1958 to 1978, 3 607 women with invasive cervical carcinoma were reported to the Stockholm-Gotland Tumour Registry. A decreased incidence rate was found among women born 1910 to 1930 when comparing the 5-year cumulative incidence rate for the different birth cohorts at the same age. A reduced mortality was demonstrated in the same way. The impact of large-scale mass screening on the incidence and mortality of cervical carcinoma is discussed. PMID- 6297249 TI - Combined effects of mining and smoking in the causation of lung carcinoma. A case control study in northern Sweden. AB - Within a case-control study of male lung carcinoma in northern Sweden combined effects of underground mining (iron ore mines) and smoking were analysed. A synergistic effect was found approximately of multiplicative type. Cases with lung carcinoma exposed to underground mining had a considerably lower average cumulative tobacco consumption than other lung carcinoma cases as an expression of the fact that smoking is particularly dangerous in underground miners. Small cell undifferentiated carcinoma was overrepresented among the cases exposed to underground mining and were especially often low tobacco consumers. In the 2 municipalities where the iron mines were located 74 per cent of the male lung carcinoma incidence could be explained by smoking and 55 per cent by underground mining (etiologic fractions). PMID- 6297251 TI - Toxicity and radiation protective effect of WR-77913 in BALB/c mice. AB - The toxicity and radiation protective effect obtained by intraperitoneal injection of WR-77913 have been investigated in BALB/c mice. The toxic LD50/30 was 3574 mg/kg. When WR-77913 was given 30 min before whole body irradiation adequate protection was observed against bone marrow and gut death. Dose modifying factors of 1.91 and 1.76 for bone marrow and 1.95 and 1.80 for gut death were obtained with drug doses roughly equivalent to one half and one quarter of the maximum tolerable doses (MTD). No protective effect was observed against central nervous system injury. Preliminary experiments with the EMT6 tumor show limited tumor protection with a dose of WR-77913 equivalent to 70 per cent of the MTD. Because of its low toxicity, adequate bone marrow and gut protection at doses equivalent to one quarter MTD and limited protection of the EMT6 tumor, WR-77913 deserves further investigation to determine its value in multiple drug dose regimens and its capability to protect other normal tissues. PMID- 6297252 TI - Radiation resistance in mice increased following chronic application of Li2CO3. AB - In experiments on strain H mice the increased radiation resistance of mice was analysed after three weeks' feeding with a diet including Li given as lithium carbonicum. The concentration of Li in the serum during the first three days of feeding was increased to 0.5 mmol/l and remained at that level to the end of feeding. The application of Li increased the overall number of stem cells in the spleen by 80 per cent compared with the control group. D0 of the line of dependence of the number of endogenous colonies on radiation dose increased following Li application by 1.2 Gy compared with controls. The proliferation activity of haemopoietic stem cells observed 90 min after injection of hydroxyurea was, after 21 days feeding with a mixture containing Li, increased by 200 per cent. The results support the idea that the increased radiation resistance of mice following feeding with Li salts before irradiation may be due to the increased content and resistance of the haemopoietic stem cells, as well as activation of granulopoiesis. PMID- 6297253 TI - Growth kinetics of Bp8 mouse ascites sarcoma after single doses of whole body irradiation. II. Analysis of the progression of cells through the cell cycle. AB - Bp8 ascites sarcoma cells growing in vivo were whole body irradiated with doses of 1.75 to 8 Gy. The inflow and outflow rates of cells in the various compartments of the cell cycle were estimated on the basis of sequential analysis of the total number of cells and from the proportion of cells in G1, S-phase, G2 and M. The flow through the compartments was calculated from these data and from the sizes of the cell pools. The durations of the mitotic delay, the G1-depletion and the early G2 blockage were linearly related to the dose. After release of the mitotic division delay the accumulation rate of cells in M and G1 decreased linearly with the dose; for G2, plateau values for the accumulation rate were found after 2.5 Gy. The outflow rates from G2 and M after release of the G2 blockage showed a shoulder type of dose-response with a D0 of 2.5 Gy and a n value of 1.5. In addition to an increase in the duration of G2, generally responsible for the increase in the total cell cycle time at about 24 hours after irradiation and an increase in the duration of the S-phase up to 12 hours after irradiation, an early increase in the duration of G1 and M was observed. PMID- 6297254 TI - Enhancement of concomitant immunity after radiation therapy and immunotherapy in a syngeneic murine tumour system. AB - Concomitant immunity was evaluated in vivo towards a chemically induced epidermoid carcinoma transplanted in a syngeneic situation. Radiation therapy reversed the declining phase of concomitant immunity associated with tumour progression. The rejection rate of a challenge graft amounted to 82 and 95 per cent, respectively, 2 and 8 days after 25 Gy as compared with 55 and 43 per cent in unirradiated controls. Radiation induced immune recovery was dose related and proved to be different from restoration of immunity following surgical removal of the tumour. Immunotherapy with intraperitoneal injection of Corynebacterium parvum significantly improved concomitant immunity. Immunostimulation and irradiation at low dosage act synergistically on host anti-tumour resistance. PMID- 6297255 TI - Mammalian spermatogenesis as a new system for biologic dosimetry of ionizing irradiation. PMID- 6297256 TI - Starch microsphere induced small intestinal ischaemia. Blood flow and morphologic investigations of late effects. AB - Severe temporary small intestinal ischaemia was induced in 15 cats by mesenteric embolization of degradable starch microspheres. After an observation period of 14 days, small intestinal histopathology and tissue blood flow were investigated. Intracellular starch fragments were found in tissue macrophages in ileal Peyer's patches in all animals, but no late adverse effects of starch microsphere embolization were observed. PMID- 6297257 TI - Liquid ionization chamber for absorbed dose determinations in photon and electron beams. AB - The use of a liquid ionization chamber for measurements in electron and photon beams with energies above 1 MeV has been investigated. The liquid in the chamber is 2,2,4-trimethylpentane. Fundamental properties such as the reproducibility of the measured charge per absorbed dose, the temperature dependence, and the angular dependence have been experimentally analysed. The general recombination losses in pulsed beams are shown to be dependent on the transport time of the ions in the liquid. For a longer transport time than the time between two consecutive pulses the losses depend on the mean dose rate and for a shorter transport time the losses depend on the absorbed dose per pulse. The Jaffe method is used for determination of the initial recombination losses, and compared with results from other authors. The chamber is particularly useful for absorbed dose determination in electron beams since the relation between measured charge and absorbed dose to water is almost energy independent. In photon beams the relation is somewhat quality dependent particularly at qualities below 10 MV. The change of the free ion yield caused by the small change of mean LET with the quality of the photon beam is probably the reason. Procedures for the calibration of the liquid chamber and the absorbed dose determination in electron and photon beams are described. PMID- 6297258 TI - Enzyme histochemical study on bone tumors. AB - A total of 19 cases with bone tumors, including six osteosarcomas. three giant cell tumors of bone, one malignant fibrous histiocytoma, four nonossifying fibromas, four chondromas and one chondrosarcoma, were examined as to enzyme histochemistry; the enzymes consisted of alkaline phosphatase (ALPase), acid phosphatase (ACPase), nonspecific esterase (NSE), adenosine triphosphatase (ATPase), 5'-nucleotidase (5'-Nucl) and beta-glucuronidase (beta-Gl). Osteosarcoma was strongly positive for ALPase followed by 5'-Nucl. Giant cell tumor, malignant fibrous histiocytoma and nonossifying fibroma showed enzyme histochemistry similar to each other: multinucleated giant cells and round cells in these tumors were strongly positive for ACPase, NSE, ATPase and 5'-Nucl simulating osteoclasts and histiocytes, whereas spindle cells were positive for ATPase and 5'-Nucl in their cytoplasm and weakly positive for ACPase. Chondroma and chondrosarcoma were focally positive for ACPase and NSE; the ACPase was sensitive to tartaric acid treatment. These observations showed that ALPase activity is very characteristic to osteosarcoma, and is useful for its diagnosis. From enzyme histochemistry, giant cell tumor, malignant fibrous histiocytoma and nonossifying fibroma can be regarded as a histiocyte-derived tumor of bone in contrast to osteosarcoma and cartilaginous tumors. PMID- 6297259 TI - Pretreatment serum albumin concentration and lactic dehydrogenase activity as prognostic factors in patients with small cell lung cancer. AB - Pretreatment laboratory parameters were analyzed as prognostic factors in patients with small cell lung cancer. Serum lactic dehydrogenase activity, serum albumin concentration, PPD skin reaction, and peripheral lymphocyte count were of prognostic importance. When these factors were evaluated by multivariate analysis together with performance status and disease extent, lactic dehydrogenase and albumin were the most influential factors related to survival. PMID- 6297260 TI - Methemoglobin formation by paraquat. AB - Paraquat is a broad spectrum herbicide known to be highly lethal to man and animals. Its toxicity is characterized by acute lung injury. Paraquat produces such toxic effects through the generation of the superoxide anion according to one proposed mechanism. The present experiment, methemoglobin formation was demonstrated after incubation of oxyhemoglobin with paraquat. The generation of the superoxide anion through the interaction of oxyhemoglobin with paraquat was suggested by chemiluminescence of luminol. Superoxide dismutase (SOD) and catalare inhibited methemoglobin formation. The generation of the superoxide anion is discussed in regard to methemoglobin formation by paraquat. PMID- 6297261 TI - Herpesvirus antibodies and vascular complications in essential hypertension. AB - Antibodies against herpes simplex virus (HSV) and cytomegalovirus (CMV) were examined in sera from 132 patients with essential hypertension and 54 normotensive healthy subjects of the same age and sex. Prevalences of HSV and CMV antibodies (titre greater than or equal to 4) were equal in patients and controls. A HSV antibody tire greater than or equal to 64 was found in 39.5% (17/53) of patients with WHO stage III hypertension, in 26.2% (22/85) of patients with stage I-II hypertension, and in only 9.4% (5/54) of normotensive subjects (p less than 0.0005). The HSV antibodies were mainly of type 1. No association between CMV antibodies and vascular complications could be demonstrated. PMID- 6297262 TI - Clinical studies on phosphate handling in hypercalcaemia. AB - Phosphate indices (serum phosphate, tubular reabsorption of phosphate, renal threshold phosphate concentration (TmP/GFR) and index of phosphate excretion) were studied in 88 hypercalcaemic subjects: 64 with primary hyperparathyroidism (HPT) and 24 with hypercalcaemia from other causes, predominantly malignant disease. HPT patients as a group could easily be separated from normal subjects (n = 16) and patients with functional hypoparathyroidism (n = 7) by use of the phosphate variables but these indices were of little discriminating value for the differential diagnosis between HPT and hypercalcaemia from other causes. There was no difference in the urinary cyclic adenosine monophosphate (cAMP) excretion between the two hypercalcaemic patient groups, but HPT patients had clearly elevated serum parathyroid hormone (PTH) levels compared with normal PTH concentrations in patients with other causes of hypercalcaemia. A positive correlation between cAMP and serum calcium and an inverse relationship between cAMP and TmP/GFR were found in patients with hypercalcaemic malignant disease. These findings suggest the existence of a humoral factor with PTH-like effects in malignant disease. Since PTH levels were low, the physiological actions were apparently not mediated by circulating PTH. No difference in the values for phosphate variables, PTH, cAMP or serum calcium was found between renal stone forming and stone-free patients with HPT. PMID- 6297263 TI - Remission by chemotherapy of the Eaton-Lambert myasthenic syndrome in a patient with small cell bronchogenic carcinoma. AB - A 70-year-old man developed symptoms compatible with the Eaton-Lambert syndrome, and a small cell bronchogenic carcinoma was diagnosed. During treatment with cytostatics his muscular strength was restored and the lung tumour as well as the electrophysiologic findings characteristic of the Eaton-Lambert syndrome disappeared. To our knowledge, it has not been reported before that chemotherapy alone of a small cell bronchogenic carcinoma has resulted in clinical, roentgenologic, and electrophysiologic remission of the Eaton-Lambert syndrome. PMID- 6297264 TI - Adrenergic receptors: classification, ligand binding and molecular properties. AB - The interaction of catecholamines and drugs with adrenergic receptors leads to a set of biochemical reactions which ultimately results in a physiological response. A brief review is given of the classification of adrenergic receptors into subtypes and the use of ligand binding techniques for the identification and characterization of these receptors. Recent advances in the biochemistry of adrenergic receptors are reviewed with special reference to the interaction of the beta and alpha 2-receptors with guanine nucleotide regulatory proteins and adenylate cyclase. The role of calcium and phosphoinositides in the function of the alpha 1-receptor is also discussed. PMID- 6297265 TI - Functionally distinct subpopulations of human monocytes: receptors for F-Met-Leu Phe are expressed only on the chemotactically responsive cells. PMID- 6297266 TI - Modulation of granulocyte response to the chemoattractant F-Met-Leu-Phe. PMID- 6297267 TI - Pulmonary alveolar macrophage chemotaxis in malignant tumors of the lung. PMID- 6297268 TI - Surface properties of activated macrophages: sensitized lymphocytes, specific antigen and lymphokines reduce expression of antigen F4/80 and FC and mannose/fucosyl receptors, but induce Ia. PMID- 6297269 TI - Regulation of macrophage functions by interferon. PMID- 6297270 TI - Macrophage activation and effector mechanisms against microbes. AB - The term activation is used to designate biochemical and functional changes that are induced in macrophages by a variety of stimuli, including interaction with microbial products, synthetic substances, immunoglobulins of different classes, and factors released by lymphocytes. The changes observed comprise an increased capacity to destroy intracellular microorganisms and non-microbial target cells as well as the stimulation of biochemical pathways leading to the release of enzymes and the generation of various toxic compounds. Activation may thus be viewed as a process aimed at recalling those metabolic functions that are necessary for killing, when phagocytosis has failed to evoke them. The increased microbicidal capacity of activated macrophages is linked to the production of oxygen intermediates, as illustrated by the study of macrophage toxicity for certain intracellular protozoan parasites. Scavengers of oxygen metabolites inhibit parasite killing in macrophages; on the contrary, agents that stimulate the production or utilization of such intermediates enhance the microbicidal effect of phagocytes. Several mechanisms enable microorganisms to survive within macrophages. In some instances, intracellular survival appears to depend on the capacity of microorganisms to be endocytized without awakening the host cell oxidative machinery. In addition, the endowment of microorganisms in endogenous enzymatic scavengers of oxygen metabolites may play a role in promoting intracellular survival. These and other mechanisms, such as the property to avoid the harmful effects of lysosomal constituents by inhibiting phagosome-lysosome fusion, or by releasing agents that block the lysosomal enzymatic machinery, may explain why certain microbes are able to survive within activated macrophages. PMID- 6297271 TI - Release of superoxide and hydrogen peroxide from guinea-pig alveolar macrophages during phagocytosis of Mycobacterium bovis BCG. PMID- 6297272 TI - Sequential regional hyperthermia: a possible answer for the treatment of cancer. AB - A therapeutic rationale for the treatment of metastatic cancer is presented which incorporates the use of deep regional hyperthermia of major body areas such as the thorax and abdomen. These body areas would be treated sequentially with the goal of eliminating the problems inherent to whole-body hyperthermia. To achieve this goal, a tri-modality approach is considered which combines regional deep heating with regional x-ray therapy and with drug therapy using agents which are heat as well as radiation hypoxic cell sensitizers (Nitroimidazoles or glucose analogues). The rationale for such an approach is well founded at the basic research level. The benefit of the tri-modality approach is that therapeutic levels of drug, X-rays and heat can be reduced significantly and hopefully be non toxic. PMID- 6297274 TI - Strategies of regulation: signals, receptors and effectors. PMID- 6297273 TI - Effects of Rous sarcoma virus on the differentiation of chick and quail neuroretina cells in culture. PMID- 6297275 TI - Regulability of gene expression and differentiation during myogenesis. PMID- 6297276 TI - Genes coding for vimentin and actin in mammals and birds. PMID- 6297277 TI - The unusually long rat crystallin alpha A2 mRNA is monocistronic. PMID- 6297278 TI - Cell deletion in differentiation. PMID- 6297279 TI - The structure and expression of the haemoglobin genes. PMID- 6297280 TI - The treatment of early cancer of the breast. PMID- 6297281 TI - Soft tissue sarcomas: behavior and management. PMID- 6297282 TI - Advances in tracheal surgery. PMID- 6297283 TI - [Metabolic abnormality of the senile cataract report II, the Na+-K+-ATPase in relation to the ATP level in the senile cataract]. PMID- 6297284 TI - Effects of vitamins B1, B2, B6 and C on erythrocyte enzymes in South African Bantu. AB - Erythrocyte transketolase (ETK), glutathione reductase (EGR) and glutamate pyruvate transaminase (EGPT) enzyme activities and coenzyme effects (in vitro coenzyme stimulation) were studied in 30 random, 10 primary hepatoma, and 3 pellagrins natives from Mozambique. Twenty-nine subjects of the random group exhibited ETK coenzyme effects below 20%. Urinary thiamine levels in this group were in normal or high ranges. The primary hepatoma group had 4 with ETK coenzyme effects above 30%, and 3 of the 4 had low level urinary thiamine excretions. Of the three pellagra patients in the study, none showed biochemical vitamin B1 deficiency. All primary hepatoma and 23 random natives had EGR coenzyme effects above 30%, but the daily urinary riboflavin excretions correlated with the coenzyme effect in only the random group. EGPT activities were spread over a wide range. The random group with high EGPT activity showed a correlation with low coenzyme effect, while the primary hepatoma group did not exhibit this correlation. After 4-day single-vitamin treatment, vitamin B1 increased total ETK and vitamin B2 increased total EGR (after in vitro coenzyme saturation). Vitamin B6 did not increase total EGPT. Vitamin B2 was less effective on total EGR in primary hepatoma than in random subjects. PMID- 6297285 TI - Congenital cytomegalovirus infection: a significant cause of deafness and mental deficiency. PMID- 6297286 TI - Constipation in the elderly. AB - Constipation is a frequent complaint among the elderly. Misconceptions about normal function and insistence on a daily bowel movement may be factors which have led to the overuse of laxatives. Evidence of gastrointestinal blood loss, a recent change in bowel elimination pattern or complications of constipation are indications for evaluation by complete laboratory, endoscopic and radiographic procedures. Natural fiber, careful attention to diet and fluid intake, regard for toilet needs and encouragement of activity are recommended as effective means for correcting constipation. PMID- 6297287 TI - Antiviral agents. PMID- 6297288 TI - Chemical interference effects in the measurement of atmospheric toluene diisocyanate concentrations when sampling with an impregnated paper tape. PMID- 6297289 TI - NIOSH lifting guidelines. PMID- 6297290 TI - Quantitation of aortic and mitral regurgitation in the pediatric population: evaluation by radionuclide angiocardiography. AB - The ability to quantitate aortic (AR) or mitral regurgitation (MR), or both, by radionuclide angiocardiography was evaluated in children and young adults at rest and during isometric exercise. Regurgitation was estimated by determining the ratio of left ventricular stroke volume to right ventricular stroke volume obtained during equilibrium ventriculography. The radionuclide measurement was compared with results of cineangiography, with good correlation between both studies in 47 of 48 patients. Radionuclide stroke volume ratio was used to classify severity: the group with equivocal regurgitation differed from the group with mild regurgitation (p less than 0.02); patients with mild regurgitation differed from those with moderate regurgitation (p less than 0.001); and those with moderate regurgitation differed from those with severe regurgitation (p less than 0.01). The stroke volume ratio was responsive to isometric exercise, remaining constant or increasing in 16 of 18 patients. After surgery to correct regurgitation, the stroke volume ratio significantly decreased from preoperative measurements in all 7 patients evaluated. Results from the present study demonstrate that a stroke volume ratio greater than 2.0 is compatible with moderately severe regurgitation and that a ratio greater than 3.0 suggests the presence of severe regurgitation. Thus, radionuclide angiocardiography should be useful for noninvasive quantitation of AR or MR, or both, helping define the course of young patients with left-side valvular regurgitation. PMID- 6297291 TI - Pectin digestion in humans. AB - The digestibility of pectin, a component of dietary fiber was investigated in humans. The groups studied comprised healthy people with intact gastrointestinal tracts, and patients who had undergone total colectomy followed by ileostomy in the management of ulcerative colitis. The pectin content of the individual plant foods in the diet and the pectin content of the excreta were determined. Some loss of pectin occurred in the small intestine but most of the pectin was degraded in the large intestine. PMID- 6297292 TI - Isoantigen status in condyloma acuminata of the uterine cervix: an immunoperoxidase study. AB - The immunoperoxidase technic was used to investigate the blood isoantigen status in condyloma acuminata, which is regarded as being caused by the human papillomavirus (HPV). Since HPV is associated with epithelial atypias and intra epithelial neoplasia, and since epithelial malignant transformation is associated with isoantigen loss, the purpose of the study was to determine if koilocytotic atypias are associated with isoantigen loss. Complete isoantigen loss was seen in 33% of cases, partial loss in 47%, and retention in 20%. The significance of this study lies in being able to recognize those lesions that may be associated with malignant transformation (80%) as indicated by isoantigen loss. Isoantigen retention may identify those epithelial atypias that undergo spontaneous regression. Long range follow-up of such patients will help further elucidate the role of HPV in neoplastic transformation of condylomatous lesions. The immunoperoxidase technic can be used in retrospective studies of condylomata of the cervix. PMID- 6297294 TI - Russell bodies--immunoglobulins? PMID- 6297293 TI - Disseminated Mycobacterium avium-intracellulare infection in homosexual men with acquired cell-mediated immunodeficiency: a histologic and immunologic study of two cases. PMID- 6297295 TI - Granular cell tumor of the stomach: a case report and review of the literature. AB - A case of granular cell tumor of the stomach in a 39-yr-old black female with peptic ulcer symptoms is presented. This lesion, a soft tissue tumor of disputed origin, has been reported in the stomach only 17 times before. In each case, the tumor was benign. Most of the patients were black, middle aged, and manifested symptoms of peptic ulcer disease. For this reason, we suggest that granular cell tumor be given serious consideration in the differential diagnosis should an upper gastrointestinal contrast study reveal a submucosal, benign mass lesion in a patient with these characteristics. PMID- 6297296 TI - Prevalence of antibody to hepatitis A virus in Okinawa and Kyushu, Japan. AB - Between 1968 and 1981, a total of 1955 serum samples from healthy subjects chosen at random in seven districts of Okinawa and two districts of Kyushu were surveyed for antibody to hepatitis A virus (anti-HAV) by radioimmunoassay. Overall prevalence of anti-HAV was 55.1% in Okinawa and 35.9% in Kyushu. Prevalence of less than 10% was observed in subjects less than or equal to 14 years of age in Okinawa and less than or equal to 24 years of age in Kyushu. In three of the districts of Okinawa, second serum samples were collected after intervals of eight, 10, and 12 years, respectively. Overall prevalence of anti-HAV decreased significantly over these time periods. When the age-specific prevalence of anti HAV on the first occasion is compared with that on the second occasion, it can be seen that there have been few new cases of hepatitis A infection. These data suggest that hepatitis A infection among children has declined dramatically in recent years, and that young people may be highly susceptible to hepatitis A virus. PMID- 6297297 TI - Kinetic alterations of the red cell membrane phosphatase in alpha- and beta thalassemia. AB - We studied the red cell membrane neutral phosphatase, which is part of the Na+K+ ATPase, in several types of oxidative hemolytic anemias. We used an artificial substrate, the p-nitrophenylphosphate. In controls and in patients heterozygous for various unstable hemoglobins (Hb Hope, Hb Koln, or Hb Hammersmith), the kinetics were of the Michaelis-Menten type. On the contrary, in nearly all patients with alpha- or beta-thalassemia, the kinetics displayed an abnormally biphasic character. The apparent Michaelis constant (KMapp) was significantly decreased. The biphasic character correlated with the imbalance of globin chain synthesis. The beta-mercaptoethanol markedly increased Vmax in controls, but had little effect on the biphasic kinetics. Omission of K+ abolished the biphasic kinetics. The abnormal kinetics failed to appear with another artificial substrate, the 4-methylumbelliferylphosphate, nor did it appear with ATP, the natural substrate. In vitro, H2O2 treatment of normal and thalassemic red cells was unable to induce or exaggerate, respectively, the biphasic kinetics, but generated alterations of a different nature. We suggest that the various kinetic alterations of the phosphatase in thalassemic syndromes originate from the imbalance of globin chain synthesis. However, the involvement of an oxidative process remains to be demonstrated. PMID- 6297298 TI - Enzyme alterations in leukemic cells. AB - Recently, much attention has been focused on various enzyme alterations found in leukemic cells. Most of the data generated thus far has involved the study of terminal deoxynucleotidyl transferase, the purine pathway enzymes, and hexosaminidase and other lysosomal enzymes. Differences in both total enzyme activities and isoenzyme patterns have been found to occur among the various leukemia types and subtypes. These changes may prove to be useful aids in diagnosing, classifying, detecting subclinical recurrent or residual disease, and as therapeutic determinants in hematopoietic neoplasia, especially in the lymphoid malignancies. PMID- 6297299 TI - A sib-pair strategy for the use of restriction fragment length polymorphisms to study the mode of transmission of type II diabetes. AB - Three models for explaining the joint distribution of DNA insertional elements and type II diabetes in sibships are given as potential candidates for resolving the mode of transmission for this common non-Mendelian disorder. While the distributions predicted from all models are subject to the same rigid marginal constraints, only two models can be considered genetic (a combined major locus multifactorial background model and a two-locus model). A sequential probability ratio test is proposed to distinguish between these two models. Once the probands have been ascertained and using realistic parameter estimates, it is shown that, on the average, fewer than 100 affected sib-pairs are required to reach a decision at the 1% significance level with 99% power. PMID- 6297300 TI - Mediators of postreceptor action of insulin. AB - Intracellular insulin action has been investigated in terms of the control of glycogen synthesis. The action was systematically traced back to the initial action of insulin at the cell membrane. Insulin activates the rate-controlling enzyme, glycogen synthase. The mechanism of this activation was found to be the control of glycogen synthase by covalent phosphorylation and dephosphorylation. Insulin brought about dephosphorylation and enzyme activation, which led to increased glycogen synthesis. A study of the interconversion reactions led to the discovery of the cyclic adenosine monophosphate dependent (cAMP-dependent) protein kinase which phosphorylates glycogen synthase and inactivates the enzyme, and a phosphoprotein phosphatase which dephosphorylates glycogen synthase and activates it. Studies revealed that insulin does not act on glycogen synthase by decreasing basal tissue concentrations of cAMP (or by altering cyclic guanosine monophosphate (cGMP) tissue concentrations); rather, insulin acts more directly on the cAMP-dependent protein kinase to inactivate the kinase and to desensitize it to the activating action of cAMP. An intracellular mediator of insulin action was hypothesized to carry out this effect on the kinase; therefore, the kinase was used as an assay to search for the putative mediator. Such an insulin generated mediator was found to be present initially in skeletal muscle and more recently in several insulin-sensitive tissues. Present evidence, although indirect, strongly suggests (1) that the mediator is a small peptide or peptide like molecule, (2) that probably several mediators (or a family of mediators) are formed rapidly with insulin action; and (3) that they are formed from cell membrane proteins by a process of limited proteolysis, which is initiated by the binding of insulin to its receptor. The present status of the rapidly developing area of hormone-induced transmembrane signaling via mediators is reviewed. PMID- 6297301 TI - Prenatal screening of pregnant women for infections caused by cytomegalovirus, Epstein-Barr virus, herpesvirus, rubella, and Toxoplasma gondii. AB - A service-oriented screening program for serologic detection of viral and parasitic infections during gestation is described for three different population groups. Significance of a determination of primary cytomegalovirus infection is documented. A screening program for detection of immunity to rubella is recommended, but the feasibility of similar programs for herpes simplex virus and Epstein-Barr virus warrants further study. Because of the low frequency of toxoplasmosis, education of patients instead of screening may be appropriate. PMID- 6297302 TI - Qualitative and quantitative assessment of the circadian rhythm of cortisol in pregnancy. AB - The effect of pregnancy on the circadian rhythm and diurnal excursion of plasma cortisol and urinary free corticoids was examined in a sequential study during the second and third trimester and 6 to 12 weeks post partum. Hourly blood samples from six subjects and 8-hour urine collections from eight subjects were obtained around the clock. While the circadian rhythm was maintained during gestation, plasma cortisol levels (24-hour mean, nadir, peak, and nadir-peak excursion) increased. The relative excursion of plasma cortisol (expressed as the percentage of deviation from the 24-hour mean) exhibited remarkable blunting compared with postpartum values. This pregnancy-associated blunting of plasma cortisol excursion was indicated by a significant reduction in the: (1) mean peak and nadir excursion, (2) integrated area between the percent deviation curve and the 24-hour mean, and (3) mean slope of the major incremental and decremental segments of the percent deviation curve. The circadian rhythm and diurnal excursion of plasma cortisol were reflected in urinary free corticoid values. Mean 24-hour urinary free corticoid concentrations increased 180% during gestation over nonpregnant levels. Nadir concentrations of urinary free corticoids in pregnancy exceeded peak nonpregnant levels. The gestational rise of metabolically active free cortisol and adrenocorticotropin (ACTH), and the pregnancy-associated blunting of the excursion of plasma cortisol may be explained by an autonomous source of ACTH during gestation. PMID- 6297303 TI - Modulation of angiotensin II pressor responsiveness by circulating levels of angiotensin II in pregnant sheep. AB - During human and ovine pregnancy, systemic pressor responses to infused angiotensin II are decreased when compared to the nonpregnant state. An increased circulating level of angiotensin II has been proposed as one of the possible mechanisms responsible for this refractoriness. The present study was designed to determine if increased pressor responsiveness to exogenous angiotensin II occurs after circulating levels of angiotensin II in pregnant ewes are reduced. Pregnant (112 to 118 days' gestation) and nonpregnant sheep were instrumented with catheters in the femoral artery and vein. Dose-response curves to angiotensin I, angiotensin II, and norepinephrine were established prior to and during continuous short-term (2 to 3 hours' duration) and long-term (24 hours' duration) inhibition of angiotensin converting enzyme activity by either captopril or enalapril. Short-term infusions of converting enzyme inhibitors blocked the formation of angiotensin II from exogenously administered angiotensin I but did not alter pressor responsiveness to angiotensin II or norepinephrine in pregnant and nonpregnant sheep. Long-term infusions of the angiotensin converting enzyme inhibitor enalapril blocked responses to angiotensin I but did not alter pressor responses to norepinephrine in pregnant and nonpregnant sheep. In contrast, responses to angiotensin II were significantly potentiated in pregnant sheep but were not altered in nonpregnant sheep. These results suggest that increased circulating levels of angiotensin II in pregnant sheep are responsible for pregnancy-induced decreases in angiotensin II responsiveness. PMID- 6297304 TI - Two cases of uveal metastasis from breast carcinoma in men. AB - We studied two cases of uveal metastasis from carcinoma of the breast in men. One patient was a 51-year-old man who developed ocular symptoms nine months after excision of a breast mass histologically diagnosed as an infiltrating ductal carcinoma. After the patient committed suicide, an autopsy disclosed metastatic lesions to the lungs, mediastinum, and chest wall. The second patient was a 49 year-old man who complained of ocular symptoms three years after undergoing mastectomy for breast carcinoma. One month after the onset of the ocular symptoms, his left eye was enucleated. Metastatic lesions were detected in the lungs and brain. More than one year after undergoing bilateral orchiectomy, and after whole-brain radiation therapy (4,500 rads) and chemotherapy, the patient has no neurologic symptoms. It appears that mammary carcinoma in men has a poorer prognosis than it does in women, probably because of two main factors: the advanced clinical stage of the disease when it is first detected and the more diffuse infiltrating character of the mass because of the scarcity of mammary tissue in men. PMID- 6297305 TI - Expression of a neural type of intermediate filament as a distinguishing feature between oat cell carcinoma and other lung cancers. AB - Expression of intermediate filaments was studied immunohistologically in oat-cell (6 cases), epidermoid (9 cases), adeno- (7 cases), large-cell anaplastic (3 cases), and bronchioalveolar carcinoma (3 cases), of lung. Affinity-purified antibodies against epithelial (anti-keratin), neural (anti-neurofilament), muscle (anti-desmin) and mesenchymal (anti-vimentin) intermediate filaments were used. In indirect immunofluorescence microscopy of oat-cell carcinomas a positive cytoplasmic fluorescence was seen only with antibodies against neural intermediate filaments, neurofilaments, while no staining of the tumor cells was observed with antibodies against other types of intermediate filaments. On the other hand, all the epidermoid, adeno, anaplastic, and bronchioalveolar carcinomas showed constantly a strong reaction with anti-keratin antibodies in a varying number of cells but no decoration with anti-neurofilament antibodies. The results show that expression of neural intermediate filaments is a major distinguishing feature between oat-cell carcinomas and other lung cancers and suggest that anti-neurofilament antibodies can be used as a diagnostic aid in the surgical pathologic study of pulmonary neoplasms. PMID- 6297307 TI - Brown adipose tissue in genetically obese (fa/fa) rats: response to cold and diet. AB - Young genetically obese (fatty, fa/fa) rats (7-8 wk old) maintained on a chow diet at 28 degrees C have a relatively normal amount of brown adipose tissue (BAT) (normal protein content, normal noradrenaline content, normal or slightly reduced cytochrome oxidase content, 30% reduction in DNA content) with cells grossly hypertrophied by accumulation of lipid. The binding of purine nucleotides by BAT mitochondria is lower in fa/fa rats than in lean rats, suggesting a lesser thermogenic activation of this tissue. Acute exposure to cold (24 h at 4 degrees C) activates BAT thermogenesis (visible hyperemia, marked increase in mitochondrial binding of purine nucleotides, depletion of noradrenaline content) in fa/fa rats as in lean rats. In contrast, feeding a cafeteria diet to young fa/fa rats fails to activate BAT (no increase in mitochondrial binding of purine nucleotides) as it does in lean rats, and these rats accumulate more extra fat (increase in weight of gonadal white adipose tissue) than do cafeteria diet-fed lean rats. It is concluded that the young fa/fa rat has normal cold-induced nonshivering thermogenesis in BAT but defective diet-induced thermogenesis in BAT and that the consequent reduction in energy expenditure, coupled with hyperphagia, contributes to the development of its obesity. The most probable location for the defect is suggested to be associated with the hypothalamus. PMID- 6297306 TI - Macrophage polykaryon formation in vitro by peritoneal cells from mice given injections of sodium periodate. AB - Peritoneal macrophages from mice that have received two separate intraperitoneal injections of the sterile, soluble oxidant NaIO4 form macrophage polykaryons (MPs) in vitro, but peritoneal macrophage from untreated, peptone-treated, or mice infected with bacille Calmette-Guerin (BCG) do not. The polykaryons are noted after 18-24 hours of culture and continue to form over a 60-72-hour period. The MPs do not form if the macrophage density is less than 4 x 10(3)/sq mm. The polykaryons appear in vitro only in cultures with less than or equal to 1-5 ng/ml lipopolysaccharide (LPS) (amounts of LPS that commonly contaminate culture medium and serum). Hydrocortisone hemisuccinate (2.6 x 10(-9) M) inhibits MP formation in vitro. Lymphocytes do not influence the polykaryon formation, and supernatants from MP cultures do not cause fusion of other macrophages. Microcinephotography demonstrates fusion of the macrophages to form the large polykaryons, which are less motile than uninuclear macrophages. The polykaryons assume different forms; generally, the nuclei (mean, 16.8 nuclei/MP; range, 2-137 nuclei/MP) are centrally located, and the nuclear chromatin of all nuclei appears similar. The MPs phagocytize polystyrene spheres and glutaraldehyde-treated erythrocytes to the same degree as do uninuclear macrophages when determined as particles per nucleus (phagocytic index), but their phagocytic index of IgG-coated erythrocytes is decreased. Peritoneal macrophages from mice given double injections of NaIO4 are nontumoricidal in the absence of LPS, but LPS, in amounts sufficient to inhibit polykaryon formation, renders the macrophages tumoricidal. Populations of macrophages containing MPs formed over 3 days of cultures also respond to LPS or macrophage activating factor (MAF) to demonstrate enhanced tumoricidal activity. PMID- 6297308 TI - Direct antigonadal activity of cannabinoids: suppression of rat granulosa cell functions. AB - The direct effects of delta 9-tetrahydrocannabinol (THC) and related cannabinoids on ovarian granulosa cells were studied in vitro. Granulosa cells from immature, hypophysectomized, estrogen-treated rats were cultured for 2 days in an androstenedione-supplemented medium in the presence or absence of follicle stimulating hormone (FSH) (10 ng/ml) with or without cannabinoids. FSH treatment increased progesterone and estrogen biosynthesis, whereas concomitant treatment with THC led to a dose-dependent inhibition of the FSH-stimulated accumulation of progesterone and estrogen with ED50 values of 3.5 +/- 0.3 X 10(-7) and 1.8 +/- 0.2 X 10(-6) M, respectively. Treatment with related but nonpsychoactive cannabinoids (cannabidiol, cannabinol, cannabigerol, or cannabichromene) was equally effective. The THC-induced inhibition of progesterone production was reversible and was associated with an inhibition of pregnenolone biosynthesis and a decrease of 3 beta-hydroxysteroid dehydrogenase activity. In addition, treatment with THC brought about a dose-dependent inhibition of the FSH-induced increase in luteinizing hormone (LH) receptors. The inhibitory effects of THC were not associated with changes in cell number, protein content, or cell viability. Thus, THC exerts direct inhibitory effects on FSH-dependent functions related to steroidogenesis and the acquisition of LH receptors, all of which are essential to follicular maturation. Because plasma concentrations of THC similar to those used in this study have been reported in human beings, repeated exposure of female users to THC may lead to ovarian dysfunction, due in part, to the direct antigonadal activity to THC. PMID- 6297309 TI - Nycterohemeral difference in inhibition of stress-induced ACTH in adrenalectomized rats. AB - To determine the interactions among the determinants of ACTH secretion, we examined the influence of circadian rhythmicity on glucocorticoid suppression of ACTH. Adrenalectomized rats were injected with the same amount of corticosterone at 0900 and 1800 h, and plasma ACTH concentrations were determined under basal conditions and after a standard ether stress. At 0900 h, corticosterone suppressed both basal and stress-induced plasma ACTH concentrations. At 1800 h, the same treatment suppressed basal ACTH secretion but not the stress-induced rise. Although the same amount of corticosterone was injected at both times of day, the plasma corticosterone concentration 5 min after injection was higher at 1800 h than at 0900 h. This study indicates that there is a nycterohemeral difference in feedback suppression of stress-induced ACTH secretion by a given dose of corticosterone. The daily variation in feedback inhibition may be due to the additive effect of the evening surge stimulus and the stress stimulus that together override the feedback signal. PMID- 6297310 TI - Vanadium, Na-K-ATPase, and potassium adaptation in the rat. AB - Vanadate is a potent inhibitor of Na-K-ATPase in vitro. It has been suggested that vanadium may function as a cellular regulator of Na-K-ATPase in vivo. To examine this speculation, we studied in rats the effect of high vanadate intake on 1) the tissue levels and distribution of vanadium, 2) basal activity of Na-K ATPase in various tissues, and 3) the activity of Na-K-ATPase in various organs under conditions of massive chronic potassium loading known to stimulate Na-K ATPase in the kidney and colon. Despite extremely high tissue levels of vanadium there was no demonstrable effect of the element on the basal activity of Na-K ATPase. When subjected to chronic potassium loading, rats with high tissue vanadium concentrations underwent potassium adaptation that was associated with a rise in Na-K-ATPase activity in the renal cortex, renal medulla, and colonic mucosa. Further studies are needed to support or refute the thesis that vanadium might be an intracellular regulator of Na-K-ATPase in vivo. PMID- 6297311 TI - Effect of potassium concentration on bicarbonate reabsorption in the rabbit proximal convoluted tubule. AB - The direct effects of acute changes in K+ concentration on HCO-3 (JnettCO2) and volume reabsorption (Jv) were examined in isolated perfused rabbit proximal convoluted tubules (PCT). Increasing ambient K+ concentration from 5 to 8 mM did not change JnettCO2 (94.5 +/- 16.1 vs. 98.8 +/- 17.7 pmol X mm-1 X min-1) or Jv (1.27 +/- 0.15 vs. 1.24 +/- 0.16 nl X mm-1 X min-1). In contrast, reducing ambient K+ concentration from 5 to 2 mM inhibited JnettCO2 by 22% and Jv by 29%. Reducing luminal K+ concentration from 5 to 0 mM with constant bath K+ concentration at 5 mM did not affect JnettCO2 or Jv. Further reductions in bath K+ concentration to 0.5 and 0 mM showed a similar dependence of both fluxes on K+ concentration. Half-maximum inhibition of JnettCO2 was obtained at 1.1 mM ambient K+ concentration and of Jv at 0.85 mM. At zero bath K+ concentration JnettCO2 was 6.6 +/- 2.5 pmol X mm-1 X min-1 and Jv was 0.03 +/- 0.04 nl X mm-1 X min-1. To determine whether this rate of acidification was significantly different from zero, we examined the ability of the PCT to generate tCO2 concentration gradients with zero bath K+ concentration at slow perfusion rates. The tCO2 concentration gradient generated (0.94 mM) was not different from that found when the perfusate was inserted directly into the collection pipette in the absence of a tubule (0.71 mM). These data are consistent with the view that HCO-3 reabsorption is totally dependent on the Na+-K+-ATPase pump system. PMID- 6297312 TI - Basolateral membrane properties in proximal convoluted tubules of the newborn rabbit. AB - We examined the properties of basolateral cell membranes in "immature" and "mature" proximal tubules and evaluated the possibility that changes in those membranes might contribute to the reported maturation of tubule function. Proximal convoluted tubules were dissected in connection with superficial glomeruli, crimped at both ends, and induced to swell by use of ouabain, low protein bathing medium, and/or collagenase. Tubules from adult animals swelled faster than tubules from 2- to 5- or 14- to 17-day-old animals. However, the basolateral cell membranes of tubules from adult animals were three- to fourfold larger in surface area than those from 2- to 17-day animals, so that the ratio of the swelling rates to the measured basolateral membrane areas remained constant during the period from newborn to adult. We conclude that the basolateral membranes "mature" only in their surface areas and that little change occurs in their intrinsic properties such as hydraulic conductivity or ion permeability. PMID- 6297313 TI - Hormonal modulation of glomerular function. AB - Glomeruli contain receptors for many hormones. Binding of angiotensin II (ANG II) or antidiuretic hormone (ADH) to glomerular mesangial cells elicits a contractile response. Other hormones induce synthesis of cyclic nucleotides (cAMP, cGMP). Glomeruli also synthesize several prostaglandins, renin, and ANG II. Micropuncture studies in Munich-Wistar rats have examined the effects of vasoactive drugs and hormones on the filtration process. Several vasodilators increase renal plasma flow in the dog and rat, but GFR remains relatively unchanged due to an offsetting fall in the ultrafiltration coefficient (Kf). Vasoconstrictor substances such as ANG II and norepinephrine cause declines in renal plasma flow and Kf, but GFR remains constant due to an increase in the transcapillary hydraulic pressure gradient. Antidiuretic peptides and parathyroid hormone also reduce Kf. Glomerular mesangial cells may regulate Kf by contracting and reducing glomerular capillary surface area. ANG II and ADH directly stimulate mesangial cell contraction in vitro. Other hormones appear to cause contraction by inducing local ANG II synthesis. These hormonal pathways are implicated in the pathogenesis of altered glomerular function in diverse forms of renal injury. PMID- 6297314 TI - Stimulation of pepsinogen release from isolated gastric glands by cholecystokininlike peptides. AB - Gastric glands isolated from rabbit stomach were employed to study the regulation of pepsinogen secretion by peptide hormones. Cholecystokinin octapeptide (CCK-OP) stimulated pepsinogen secretion with an ED50 of about 1 nM. Caerulein was as effective as CCK-OP but less potent (ED50, 10 nM). Gastrin (HG-17) was found to be a weak stimulus, being only about 20% as effective as CCK-OP or caerulein. Sulfation of CCK-OP and caerulein was found to be important for potency but did not alter efficacy. Peptide stimulation of pepsinogen secretion was unaffected by cimetidine, atropine, or propranolol. Combinations of peptides resulted in less than-additive responses, as did the combination of peptides with carbachol. In contrast, combination of peptides with isoproterenol resulted in additive responses. Accumulation of the weak base aminopyrine was used to measure acid formation by the gastric glands. The peptides gastrin, CCK-OP, and caerulein were found to be equally effective in stimulating acid formation. The peptide stimulation of pepsinogen secretion was inhibited by dibutyryl cGMP, whereas stimulation of acid formation was not inhibited by the cyclic nucleotide. The results indicate that gastric glands contain at least two peptide receptors distinguishable by sensitivity to dibutyryl cGMP. The peptide receptor associated with pepsinogen secretion appears to be selective for CCK relative to gastrin. PMID- 6297315 TI - Analysis of cardiac adrenergic mechanisms in hypoxic lambs. AB - Adrenergically mediated inotropic and chronotropic responses to hypoxemia were studied in 12 neonatal lambs ranging from 1 to 10 days of age. Six lambs were adrenalectomized (Adnx) and six were sham operated. Inotropic changes were assessed from measurements of left ventricular dP/dtmax under controlled hemodynamic conditions. Atropine (1 mg) was given and hypoxia produced by adding N2 to the respirator. In the sham-operated lambs, dP/dtmax increased as a function of the level of hypoxemia. With a mean arterial O2 partial pressure (PaO2) of 21.7 +/- 2.0 Torr, dP/dtmax averaged 17 X 10(2) mmHg/s above control values. Adnx did not reduce resting dP/dtmax, but comparable levels of hypoxemia elicited a much smaller increase (5 X 10(2) mmHg/s) in this group (P less than 0.02). In the sham-operated lambs, ganglionic blockade with tetraethylammonium chloride (TEAC, 100 mg) reduced the hypoxic response to similar values (6.5 X 10(2) mmHg/s). These residual inotropic responses were completely abolished by beta-adrenergic blockade with practolol (4 mg/kg). Chronotropic changes were identical in both groups (18.7 beats/min) and abolished by TEAC alone. It is concluded that the major fraction of adrenergic inotropic stimulation during hypoxemia is derived from the adrenal glands and that autonomic neural function is essential to the release process. Heart rate responses are independent of adrenal integrity. A residual source of inotropic stimulation, blocked by practolol, is derived from unidentified sources. PMID- 6297316 TI - Chemiosmotic potassium ion pump of insect epithelia. AB - Transporting epithelia of insects are unlike most vertebrate epithelia in that they lack a serosa and possess septate junctions. The Na+/K+ pump is absent and an electrogenic K+ uniport pump is present in such insect epithelia as lepidopteran midgut, dipteran salivary glands, and many Malpighian tubules. The K+ pump is located in the apical plasma membrane and pumps K+ out of the cells. In midgut the transepithelial K+ transport is against a potential difference (PD) in excess of 120 mV and against a 10-fold K+ concentration difference in vivo. The pump uses a K+-modulated ATPase thought to be located in particles called K+ portasomes, which resemble the F1-F0 ATPase of phosphorylating membranes. Like F1 F0 particles the K+ portasomes are located on the cation input, electronegative, ATP-binding side of the membrane and appear to pump two cations for each MgATP2- hydrolyzed. We propose that in K+-transporting epithelia and in phosphorylating membranes running backwards the portasomes orient the binding of ATP with respect to a cation-gated channel in such a way that when MgATP2- is hydrolyzed P-i is separated from MgADP-; the 2 K+ or 2 H+ ions are no longer neutralized and are repelled from the channel to the opposite side of the membrane. We have isolated the K+ portasome-containing goblet cell apical membrane from larval Manduca sexta midgut and are attempting to isolate the K+ portasomes and K+-ATPase. PMID- 6297317 TI - Release of tonin and of kallikrein by perfused rat submaxillary gland. AB - The release of tonin and kallikrein was investigated in the venous effluent of the perfused rat submaxillary gland (SMG). The SMG was perfused through the common carotid artery after all branches except the SMG artery had been ligated. The venous outflow was collected from the SMG vein. The contralateral nonperfused gland was used as control. The infusion of isoproterenol increased tonin release without affecting its concentration in the SMG. Theophylline increases both tonin secretion and tonin concentration in the SMG. These results suggest that beta adrenergic stimulation enhances tonin release by increasing synthesis in the SMG, and that these effects might be mediated by cAMP. The infusion of norepinephrine did not affect either tonin release or tonin concentration in the gland. Kallikrein release was significantly increased by norepinephrine, isoproterenol, and theophylline. However, whereas alpha-stimulation produced a significant decrease in kallikrein concentration in the stimulated SMG, beta-adrenergic and theophylline stimulation produced no significant changes in tissue kallikrein. These results suggest that alpha-adrenergic stimulation increases kallikrein release by depleting the enzyme from the gland, whereas beta-adrenergic stimulation might increase kallikrein release by stimulating its synthesis in the gland. PMID- 6297318 TI - Renal and adrenal responses to converting-enzyme inhibition in fetal and newborn life. AB - The renal and adrenal responses to a continuous infusion of the angiotensin converting enzyme (ACE) inhibitor captopril were studied in 27 chronically catheterized sheep fetuses (less than 120 days gestation, n = 15, and greater than 130 days gestation, n = 12; term being 145 days) and in 12 newborn lambs between 8 and 21 days of age. Total renal blood flow did not change during ACE inhibition. However, the renal vascular resistance decreased significantly in newborn lambs (-21.8 +/- 5.7%) and in fetuses greater than 130 days (-21.7 +/- 4.7%) but not in fetuses less than 120 days. A significant decrease in filtration fraction (-19.2 +/- 6.5%) was observed in newborn lambs. No changes in urinary kallikrein and prostaglandin excretion rate were observed during ACE inhibition in any group of animals. ACE inhibition produced similar declines in blood pressure in both groups of fetuses (-10.2 +/- 3% in fetuses less than 120 days and -9.5 +/- 4.6% in fetuses greater than 130 days) and in newborn lambs (-13.4 +/- 2.1%). The percent changes in plasma renin activity were similar in all groups of animals. However, a significant decline in plasma aldosterone concentration was observed only in newborn lambs (from 130 +/- 31 to 64 +/- 9 pg/ml). These results suggest that the renin-angiotensin system might have physiological significance during maturation, but that this role seems to be more important in near-term fetuses (greater than 130 days) and postnatally than early in gestation. PMID- 6297319 TI - Effects of dietary sodium restriction on heart rate control in rats. AB - The effects of dietary sodium restriction on heart rate control were evaluated in male Wistar rats maintained for 4 wk on a low- (22 meq/kg diet) or standard (141 meq/kg diet) sodium diet. In conscious low-sodium rats, atropine produced heart rate increases greater than in standard sodium animals, indicating that basal parasympathetic tone was elevated. Sodium restriction did not affect basal sympathetic tone, since similar reductions in heart rate were produced by propranolol in both dietary groups. However, sympathetically mediated tachycardic responses to sodium nitroprusside were augmented in low-sodium rats. This enhancement of sympathetic function was also observed in low-sodium animals after pithing. The increases in heart rate produced by spinal sympathetic stimulation were significantly accentuated; whereas, responses to norepinephrine were not affected by the diets. This finding is in agreement with the observation that the chronotropic responses elicited by isoproterenol in isolated atria were not altered. When neuronal influences on heart rate were blocked by combined administration of atropine and propranolol or chlorisondamine in conscious rats, the underlying intrinsic cardiac rate was found to be significantly higher in low sodium rats. In summary, the present study identifies three effects of sodium restriction on heart rate control in the rat: 1) basal parasympathetic tone is increased, 2) heart rate responses to sympathetic activation are enhanced, and 3) the intrinsic cardiac rate is elevated. PMID- 6297320 TI - Effect of polybrominated biphenyls (PBB) on developmental abilities in young children. AB - Eighteen children, ages 4 to 6 years, with known exposure to polybrominated biphenyls (PBB) in utero and/or through breast milk were administered developmental tests. These same children had exhibited low scores on a partial developmental assessment two years earlier. Current results were compared to normative test data. Findings showed: 1) PBB cohort children are within the normal range in all areas assessed; 2) An inverse relationship is noted between PBB fat level and scores on some developmental tasks. The importance of this finding for later development is unclear and, thus will bear future monitoring. PMID- 6297321 TI - Developmental abilities of children exposed to polybrominated biphenyls (PBB). AB - To investigate whether ingestion of polybrominated biphenyls has an adverse effect on the neuropsychological development of young children exposed in utero and in infancy, five tests of the McCarthy Scales of Children's Abilities were administered to a group of 19 PBB-exposed Michigan children. When the data for the exposed group were analyzed according to body burden of PBB as determined by fat biopsy, correlations ranging from -.5228 to -.3004 were found between the natural logarithms of the children's fat PBB values and their standardized scores on the developmental scales. Four of the five correlations were significant at p less than .05. Multivariate analysis of covariance confirmed the existence of a significant main effect for fat PBB level, with parental education held constant. Children with higher body burdens of PBB (greater than .100 ppm) scored significantly lower than exposed children with lower body burdens on the same four tests, and on a composite score representing overall performance. These results suggest the existence of an inverse relationship between body levels of PBB and some developmental abilities in young children. PMID- 6297322 TI - Possible effect of neonatal polybrominated biphenyl exposure on the developmental abilities of children. PMID- 6297323 TI - Environmental politics and science: the case of PBB contamination in Michigan. AB - This article examines how politics and science interacted against a background of uncertainty to shape policy in the case of environmental contamination by polybrominated biphenyls (PBB) in Michigan. In 1973, between 500 and 1,000 pounds of the flame retardant PBB were accidentally shipped and used instead of the dairy feed additive magnesium oxide, resulting in the widespread contamination of animal feeds, animals, and human food products. The contamination was initially perceived as the private trouble of a single farmer. The problem next became a public issue as public and private institutions grappled with questions of illness, safety, and disposal. To gain influence over those institutions, dissatisfied individuals and groups then turned the PBB contamination into a political controversy. The final section of the present article analyzes how science and politics interacted in: the ways bureaucratic organizations defined the three problems of contamination; the role political controversy played in redefining problems and influencing policy; and the political roles of scientists in controversies over environmental contamination. PMID- 6297324 TI - Relationship between number of human Japanese encephalitis cases and summer meteorological conditions in Nagasaki, Japan. AB - Analysis of epidemiological and meteorological data for Nagasaki Prefecture, southwest Japan, for the period 1950-1979, showed a correlation between size of epidemics of human Japanese encephalitis (JE) and weather factors. It was demonstrated that during the period 1950-1969 the epidemic size (ES) had two types of correlation: ES correlated inversely with total summer precipitation, and ES correlated directly with mean summer temperature. The fact that in the 1970s there were fewer cases of JE cannot be attributed to changes in the weather factors. Large epidemics occurred in summers with low precipitation and high temperatures, while the epidemic size was small when there was heavy precipitation and temperatures were low. The two weather factors jointly explain 0.53 of the observed variance in annual JE ES. Furthermore, they strongly correlate with each other, and total summer precipitation alone is sufficient to explain about one-half of the total variance. How these factors influence the number of JE cases is discussed in relation to the population dynamics of the vector mosquito, Culex tritaeniorhynchus. PMID- 6297325 TI - Effectiveness of allopurinol against Leishmania braziliensis panamensis in Aotus trivirgatus. AB - Orally administered allopurinol at 50 mg/kg for 21 days showed pronounced antileishmanial activity against experimentally induced lesions of Leishmania braziliensis panamensis on the nose of Panamanian Aotus trivirgatus monkeys, with complete healing in 4 of 5, although parasitologic cure was achieved in only 2 of 5. The same total daily dose of drug, given in a divided dose twice daily, resulted in complete healing in all 5, and parasitologic cure in 4 of 5 animals. Standard treatment controls receiving 40 mg/kg of antimony stibogluconate intramuscularly for 15 days showed healing in 4 of 5 monkeys. It is not known if a similar level of effectiveness would result with this dose of allopurinol in humans, since Aotus may have a different pattern of metabolic conversion of the drug to the more active riboside. PMID- 6297326 TI - Femoral neuropathy due to retroperitoneal bleeding. A red herring in medicine complicates anticoagulant therapy and influences the Russian Communist Revolution (Crown Prince Alexis, Rasputin). AB - Femoral neuropathy occurs when occult retroperitoneal bleeding impinges on the appropriate nerve roots. The syndrome involves the acute onset of groin and thigh pain with characteristic flexion and external rotation of the hip. It may mimic other conditions such as acute arterial occlusion. Thorough knowledge of the anatomy of the femoral nerve explains the clinical features and leads the clinician to suspect the occurrence of this syndrome. Three cases have been reviewed that exhibited this condition as a result of retroperitoneal bleeding, a complication of systemic heparin therapy. The hemophilia that afflicted Alexis, the Crown Prince of Russia and son of Tsar Nicholas and Tsarina Alexandra, resulted in this clinical syndrome. The consequences enabled the sinister starets, Gregory Rasputin, to become intimately involved with the royal family, influencing the response of the Tsar to the political events in Russia, thereby playing an important role in setting the stage for the 1917 Russian communist revolution. PMID- 6297327 TI - Variants of hemipelvectomy. AB - Anterior flap hemipelvectomy is used for large, fixed tumors in the area of the buttock. The anterior flap consists of skin, subcutaneous fat, fascia lata, and the mobilized femoral vessels attached to the flap. It is accomplished by dividing the profunda femoris vessels. In routine posterior flap hemipelvectomy, a long flap can be constructed, if necessary, reaching to the level of the umbilicus. The viability of the posterior flap is assured by leaving the gluteus maximus muscle attached to it. PMID- 6297328 TI - Transport characteristics of the blood--perilymph barrier. AB - The inner ear must maintain a delicate homeostasis to preserve high sensitivity to acoustic and vestibular inputs. Various experimental conditions were imposed upon chinchillas to help identify these homeostatic mechanisms for the inner ear fluids. Radioactive ions (sodium, chloride, and calcium) pass into perilymph more slowly than they pass into cerebrospinal fluid or aqueous humor. The concentration of glucose in perilymph relative to that in serum was found to be quite constant (about 45 per cent) over a wide range of serum values (130-943 mg/dl) generated by continuous intravenous infusion. The transport of albumin into perilymph was very slight after injection of the radioactive protein intravenously. Osmotic agents cause an efflux of water from perilymph into a relatively hypertonic circulating blood, resulting in a secondary elevation of perilymph osmolarity. Recent findings relating to cyclic AMP metabolism and the possible role of this second messenger in the regulation of inner ear fluid metabolism are discussed. A new hypothesis that excess cyclic AMP in the inner ear may be related to endolymphatic hydrops is presented. PMID- 6297329 TI - [Endocrine reaction pattern in the course of a one-phase tramadol-N2O combination anesthesia]. AB - The effects of Tramadol-N2O-anaesthesia on the per- and postoperative change in blood concentrations of cortisol, prolactin, thyroxine, triiodothyronine, cyclic AMP, glucagon, antidiuretic hormone, PTH-peptide (44-68), glucose, lactate, pyruvate and free fatty acids (FFA) were investigated in connection with elective orthopaedic surgery. Anaesthesia in man with Tramadol and nitrous oxide were found to be associated with a significant elevation of plasma cortisol and plasma prolactin in man. However, cortisol secretion during anaesthesia is associated with an inhibition in T4-T3 conversion. No significant alterations in plasma glucagon concentrations were observed. Generally, surgical trauma induced a significant increase in plasma cyclic AMP with intraoperative levels between 26.4 and 34.3 pmol/ml. At the end of surgery a significant fall in plasma PTH-peptide (44-68) occurred. There was also a significant change in plasma ADH levels following induction of anaesthesia. During surgery we found plasma ADH levels up to 56 pg/ml. In addition stress and surgical trauma increased blood glucose and FFA while plasma pyruvate and plasma lactate nearly remained unchanged. The data would suggest that the non-specific stress response attributed to anaesthesia may in fact be reflecting a response to relatively light anaesthesia. PMID- 6297330 TI - Antacid aspiration. PMID- 6297331 TI - Problems encountered in measuring erythrocyte pyrimidine 5'-nucleotidase activity. PMID- 6297332 TI - Fluorometric determination of selenium in nanogram amounts in biological materials using 2,3-diaminonaphthalene. PMID- 6297333 TI - Identification of cyclic guanosine monophosphate-binding proteins in Drosophila by direct photoaffinity labeling. PMID- 6297334 TI - Immobilized enzyme system for the conversion of benzo[a]pyrene to fluorescent metabolites. PMID- 6297335 TI - Continuous-flow determination of reducing sugars and sucrose in natural and industrial products with periodate oxidation and a periodate-sensitive flow through electrode. PMID- 6297336 TI - Relevance of changes in adrenoceptor characteristics after terbutaline treatment. AB - Rats were treated with terbutaline in order to specify the in vivo effects of beta 2-sympathicomimetic treatment in relation with desensitization phenomena on the level of pulmonary adrenoceptors. Maximal binding of 3H-dihydroalprenolol (3H DHA) to lung membrane preparations, binding affinity and the relative amount of beta 2-adrenoceptors were used as parameters. These were established 1, 3, and 6 days after a 10 days terbutaline treatment and compared with control values. Ten days treatment did not cause significant changes in the parameters measured. However, maximal binding was enhanced 3 and 6 days after the last treatment, binding affinity was enhanced 3 days after the last treatment, while the relative amount of beta 2-adrenoceptors was diminished 1 and 6 days after termination of the treatment. When the amount of beta 2-adrenoceptors was calculated in terms of 3H-DHA binding, an increase could be shown 3 days after the last terbutaline treatment. Therefore the direct influence of the treatment on the beta adrenoceptor population appeared to be small, dependent on the parameter used and probably not eliciting a functional desensitization. PMID- 6297337 TI - Evaluation in swine of a subunit vaccine against pseudorabies. AB - A subunit vaccine against pseudorabies virus (PRV) was prepared by treating a mixture of pelleted virions and infected cells with the nonionic detergent Nonidet P-40 and emulsifying the extracted proteins incomplete Freund's adjuvant. Three 7-week-old pigs without antibodies against PRV were given 2 IM doses of this vaccine 3 weeks apart. Thirty days after the 2nd vaccination, 10(6) median tissue culture infective doses (TCID50) of a virulent strain of PRV were administered intranasally. Tonsillar and nasal swabs were collected daily between 2 and 10 days after challenge exposure. The pigs vaccinated with the subunit vaccine were not found to shed virulent PRV. Two groups of five 7-week-old pigs vaccinated with commercially available vaccines, either live-modified or inactivated virus, and subsequently exposed to 10(6) TCID50 of virulent PRV, shed virulent virus for up to 8 days. The subunit vaccine induced significantly higher virus-neutralizing antibody titers than either the live-modified or inactivated virus vaccine. PMID- 6297338 TI - Experimental bluetongue infections in gnotobiotic lambs and kids. AB - Ten gnotobiotic lambs and 4 gnotobiotic kids inoculated with virulent bluetongue virus (BTV) including BTV international types 10, 11, 13, and 17 did not show clinical signs of bluetongue. Seroconversion was observed in all gnotobiotes and conventional control sheep inoculated with the appropriate challenge BTV. Viremia was detected in 1 BTV-infected gnotobiote. Conventional control sheep inoculated with a given BTV international serotype showed a marked febrile response with hyperemia of the nasal and buccal mucosa. PMID- 6297339 TI - Mechanical transmission of equine infectious anemia virus by deer flies (Chrysops flavidus) and stable flies (Stomoxys calcitrans). PMID- 6297340 TI - Evaluation of the microimmunodiffusion test for the detection of antibody to pseudorabies virus. PMID- 6297341 TI - Vaccination of cattle with binary ethylenimine-treated bovine leukemia virus. PMID- 6297342 TI - Isolation of virus and antibody containing immune complexes from mink with Aleutian disease by affinity chromatography of equine complement clq. AB - Affinity chromatography on immobilized equine complement Clq was used for the isolation of complement-binding immune complexes in sera of mink infected with Aleutian disease virus. Immune complexes were isolated and quantitated from 4 of 5 infected mink, as early as 2 weeks after infection and before hypergammaglobulinemia had appeared. The quantity of immunoglobulin G in these immune complexes ranged from 180 to 370 micrograms/ml serum. There were no Clq binding immune complexes found in mink which were negative for Aleutian disease antibody. Using 125I-labeled BSA-anti-BSA complexes, we demonstrated that the affinity columns bound selectively immune complexes which had formed in antibody excess, whereas immune complexes in antigen excess were not bound. By neutralization of sensitized virus with anti-mink IgG serum, non Clq-binding immune complexes were also detected, which indicates that circulating immune complexes in persistently infected mink are heterogeneous as far as their reactivity with equine Clq is concerned. PMID- 6297343 TI - Primary malignant tumors of salivary gland origin. A 52-year review. AB - Primary carcinoma of salivary glands is uncommon, comprising approximately 30 per cent of salivary gland neoplasms. The natural history of these neoplasms varies greatly, depending largely upon the cell type. Only prolonged follow-up can furnish worthwhile data, and five or even 10 years is inadequate in several cell types. Case records of 202 patients who had major or minor salivary gland carcinomas during the years 1928 through 1979 were reviewed from the files of Vanderbilt University Hospital. We studied these records with regard to site of origin, cell type, lymph node involvement, signs and symptoms, routes of metastatic spread, and survival. Microscopic sections were available for review in 188 patients (93%). Major and minor salivary glands were involved as the primary site in 74 per cent and 26 per cent, respectively. Prognostic factors are discussed, including histologic type and grade, size, lymph node involvement, vascular and perineural invasion, growth pattern (i.e., infiltrating versus pushing margins), and presence or absence of invasion outside the gland. Our 99 per cent follow-up of these patients revealed that the natural course of many of these neoplasms was characterized by long duration, repeated local recurrences, occasional metastases to regional lymph nodes, and frequent metastases to the lungs. PMID- 6297344 TI - Intestinal pseudo-obstruction as the presenting manifestation of small-cell carcinoma of the lung. A paraneoplastic neuropathy of the gastrointestinal tract. AB - A 58-year-old woman who had presented with intestinal pseudo-obstruction died 9 months later from rapidly progressive neurologic symptoms and autonomic insufficiency. Her gastric emptying had been markedly delayed and transit of markers had been slowed throughout the small bowel. A 5-hour manometric recording of the antrum and duodenum had shown absence of the normal interdigestive motor complex, which was replaced by irregular contractile activity of reduced amplitude. A small-cell carcinoma of the lung was found at autopsy. Pathologic study of the gut showed widespread degeneration of the myenteric plexus, which was infiltrated by plasma cells and lymphocytes and contained significantly reduced numbers of neurons. The extra-intestinal nervous system had neuronal loss and lymphocytic infiltrates in dorsal root ganglia. Thus, a gastrointestinal neuropathy causing intestinal pseudo-obstruction may be the presenting manifestation of a paraneoplastic syndrome associated with small-cell carcinoma. PMID- 6297345 TI - High-dose ketoconazole for treatment of fungal infections of the central nervous system. AB - Mortality and complication rates remain unacceptably high with conventional intravenous and intrathecal therapy for patients with coccidioidal meningitis and intracerebral fungal lesions. We studied the ventricular and lumbar cerebrospinal fluid penetration of ketoconazole and the responses to therapy in two patients receiving ketoconazole orally, 800 mg daily, and amphotericin B intraventricularly for meningeal and extrameningeal coccidioidomycosis. Five patients received only 1200 mg of ketoconazole: one had uncomplicated coccidioidal meningitis, three had obstructive hydrocephalus due to coccidioidal meningitis, and one had a histoplasmal brain abscess. Ketoconazole concentrations in ventricular and lumbar fluid ranged from 0.05 to 1.65 micrograms/mL 4 and 8 hours after the dose. The mean penetration of ketoconazole (+/- SD) was 1.9% +/- 0.8% for ventricular fluid and 5.4% +/- 2.6% for lumbar fluid. Ketoconazole concentrations in cerebrospinal fluid varied directly with those in serum and with cerebrospinal fluid protein content. The encouraging clinical responses, convenience, safety, and the consistent penetration of ketoconazole into obstructed and nonobstructed cerebrospinal fluid support the use of these regimens as alternatives to conventional therapy. PMID- 6297346 TI - Ketoconazole and blastomycotic osteomyelitis. PMID- 6297347 TI - [Manual digestive sutures in gastric resection and total gastrectomy]. PMID- 6297348 TI - Epidermal growth factor. PMID- 6297350 TI - Regulation of the levels of cellular transcripts by SV40 large T antigen. PMID- 6297349 TI - Molecular biology of cell division. AB - Different domains of the SV40 A gene have different functions, such as viral DNA replication, cell DNA replication, and stimulation of cellular RNA synthesis. The sequences in the SV40 A gene that are critical for the induction of cell DNA synthesis lie on the map between nucleotide 4360 and nucleotide 4001, a stretch of 360 nucleotides coding for 120 of the 708 amino acids of the large T antigen. The sequences critical for stimulation of rRNA synthesis lie on the map further downstream, between nucleotides 3827 and 3506, thus indicating that the signals for growth in size and for cell DNA replication can be dissociated. Methylation of the SV40 A gene at multiple ECoRI* sites has no effect on its expression. However, methylation of the HSV-TK gene at one single ECoRI site 70 base pairs upstream from the cap site inhibits its expression. The results indicate that methylation of genes affects their expression, but only when methylation occurs at specific sites. PMID- 6297351 TI - Molecular dissection of mhc complex and of sv40-induced surface antigen. PMID- 6297352 TI - Microinjection of cloned SV40 DNA fragments in the study of cell proliferation. AB - DNA recombinant technology and the manual microinjection technique were used to study the base sequences in the SV40 early gene coding for cell DNA replication and nucleolar activation. Sequences critical for rRNA gene activation are located between nucleotides 3826 to 3506 (0.39-0.33 m.u.). Base sequences from nucleotide 4350 to 4190 (0.49-0.46 m.u.) are required for cellular DNA replication. Major T antigen determinant is coded by a sequence extending from nucleotide 4190 to 3880 (0.46-0.42 m.u.). Considering that an increase in cell size is regularly accompanied by an increase in the cellular amount of rRNA, nucleolar genes should be a target for growth-in-size signals. Therefore, the SV40 early gene presents a domain responsible for cell growth in size, and another separate but proximate region coding for cellular DNA replication. PMID- 6297353 TI - Insertion of new genetic information into bone marrow cells of mice: comparison of two selectable genes. AB - A system for insertion of new genetic information into mouse hematopoietic cells is described. Two selectable genes were examined: herpesvirus thymidine kinase and a mutant mouse dihydrofolate reductase. The DHFR system appears to be superior in terms of the frequency and stability of gene insertion and expression in hematopoietic tissues. About 70% of mice had indirect (karyotypic) evidence of gene insertion; of these, about 60% (three of five) had stable expression of the inserted mutant DHFR. In contrast, only 13% of mice demonstrated stable karyotypic transformation by HSVtk, and of those with stable transformation five of seven showed persistent viral gene sequences in hematopoietic tissues. PMID- 6297354 TI - A possible opioid receptor dysfunction in some depressive disorders. PMID- 6297355 TI - Dual actions of morphine on the central nervous system: parallel actions of beta endorphin and ACTH. AB - In the studies described above, the intracerebral microinjection technique was used to study the actions of morphine at morphine-sensitive sites, the periaqueductal gray (PAG) and the midbrain reticular formation (MRF). In the PAG, morphine exerted dual actions: inhibitory and excitatory. In the MRF, morphine exerted an excitatory action only, indicating that the dual actions of morphine are dissociable and site specific. Following microinjection into the PAG, beta endorphin exerted an inhibitory, and ACTH an excitatory, action, i.e., each duplicated one of morphine's dual actions. These results indicated that receptors for the endogenously occurring peptides, beta-endorphin and ACTH, may play a role in morphine's potent pharmacological actions. Although these studies do not shed direct light on the physiological role of these neuropeptides and their receptors, nor on their potential roles in the functional regulation of brain (especially in diseased mental states), it may be permissible to offer some speculations. We previously proposed that beta-endorphin may be an endogenous antipsychotogen, and that a deficiency in brain beta-endorphin may underlie some forms of psychopathology. In view of beta-endorphin's biosynthetic link to ACTH, and the behavioral effects of beta-endorphin (sedated immobility) that was found to be opposite in kind to those of ACTH (fearful hyperreactivity) following administration into brain, it is possible that these two neuropeptides may have regulatory roles in maintaining a functional balance in brain. (It may be speculated that these biosynthetically linked neuropeptides served survival functions of "flight" and "freeze" in our evolutionary ancestors.) An imbalance in the bioavailability of either of the two neuropeptides, e.g., a deficiency of beta-endorphin or an excess of ACTH (perhaps due to the lack of the specific enzymes that cleave these peptides from their parent prohormone) may be an etiological factor in some forms of chronic functional disorders of the brain. PMID- 6297357 TI - Opiate receptors and opioid peptides: an overview. PMID- 6297356 TI - Metabolism of enkephalin analogues and surrogates having enhanced pharmacologic activities. PMID- 6297358 TI - Multiple opiate receptors and their different ligand profiles. PMID- 6297359 TI - Role of opioid peptides in disorders of attention in psychopathology. PMID- 6297360 TI - Endorphins in the cerebrospinal fluid of psychiatric patients. AB - In this paper we have reported the results of studies in psychiatric patient groups using the strategy of measuring opioid activity and beta-endorphin (ir) in CSF. Our findings do not lend support to the notion of excess endorphin activity in schizophrenia, but rather suggest the possibility of a decrease in endogenous opioid activity in some schizophrenic patients. In affectively ill patients our data suggest that there may be a relative change in endogenous opioid system activity across state change in manic-depressive illness. Who also found a relationship between nurses' ratings of anxiety and CSF opioid activity in depressed patients, although it is unknown whether this directly relates to the pathophysiology of this symptom, or is related to stress response. The relationship between CSF opioid activity and HPA axis activity, as reflected by urinary free cortisol excretion, supports the notion of important physiologic relationships between these systems and raises the issue of a role for the endogenous opioid system in the abnormal activation of this system in depression. Finally, the finding of increased CSF opioid activity in anorexia nervosa patients when a minimum weight coupled with data relating endogenous opioids to eating behavior raises interesting questions regarding a possible involvement of the endogenous opioid system involvement in this illness. PMID- 6297361 TI - beta-Endorphin immunoreactivity in the plasma of psychiatric patients receiving electroconvulsive treatment. PMID- 6297362 TI - Clinical and experimental studies of stress and the endogenous opioid system. PMID- 6297363 TI - Clinical pharmacology of beta-endorphin in depression and schizophrenia. PMID- 6297364 TI - Studies of beta-endorphin in psychiatric patients. PMID- 6297365 TI - beta-Endorphin administration to acute schizophrenic patients: a double blind study. PMID- 6297366 TI - [New case of rhinoentomophtoromycosis. Cure by ketoconazole]. AB - A new case of rhino-entomophtoromycosis due to Conidiobolus coronatus is added to the ten others observed in Cameroon, among the 62 african cases described. The patient, a man 27 years old, has an elephantiasis form with nasal obstruction, hypertrophy of lips, globulous forms of cheeks, giving a monstrous facies. After failure of KI and intravenous miconazole, therapeutic success was obtained with oral ketoconazole. After improvement with 400 mg daily, the doses were increased to 600 mg to obtain mycological and clinical cure with good clinical and biological tolerance; important eosinophilia related to the destruction of the fungus was observed. Restorative surgery was necessary to render a more human aspect of the monstrous lesions of face. PMID- 6297367 TI - [Vulvar bowenoid states in the young woman]. PMID- 6297368 TI - Carcinoma of the ovary, stages I and II. A prospective randomized study of the effects of postoperative chemotherapy and radiotherapy. AB - A prospective randomized trial for comparing the effects of chemotherapy and radiotherapy on survival in malignant epithelial ovarian tumours was carried out during the years 1975-1978 in 142 (90%) of a total material of 157 patients in Stage I and Stage II of this disease. Stratification was done according to the state of the tumour capsule and the type of histology found in Stages Ia and Ib. Two types of randomized treatment were given: A. Patients with no extracystic excrescences and intact tumour capsule with mucinous tumours in Stages Ia and Ib were given Melphalan or were not treated, while those with non mucinous tumours were given Melphalan or radiotherapy; B. All of the other patients with tumours in Stage I and Stage II were treated with radiotherapy alone, or were irradiated in combination with Melphalan treatment. Both of these treatment groups had about the same rates of 2- and 5-year survival. It was seen that the histological grade and the amount of residual tumour left at surgery are important prognostic factors. Postoperative treatment does not seem to improve cases of well differentiated, early mucinous tumours in Stage Ia. Acceptable results were obtained, however, irrespective of histological type, in early Stage I, moderately differentiated tumours treated with Melphalan or radiation therapy, and in well- or moderately differentiated tumours in Stage I or Stage IIa using irradiation alone or in combination with chemotherapy. Stages IIb and IIc, and all poorly differentiated tumours in Stages I and II, require more aggressive treatment. PMID- 6297370 TI - Delayed increase of 17 alpha hydroxylase activity in adrenocortical cells exposed to corticotropin. PMID- 6297369 TI - [A new aspect of the regulation of reproduction: intragonadal cybernins]. AB - The testes and ovaries secrete peptide compounds which act on the gonads themselves and can thus be defined as cybernines. Several factors have been identified. The granulosa cells secrete an ovocyte maturation inhibitor (OMI) into the follicular fluid which opposes ovocyte miosis activity before the pre ovulatory LH peak. The follicular fluid also contains the inhibitor of FSH uptake by the receptors (FSH-RBI), which appears to be implicated in the maintenance of the follicle in the immature state or in its progression towards atresia. This FSH-RBI is also found in testicular extracts. The luteinization inhibitor (LI) prevents morphological transformation of the granulosa cells into luteal cells and the secretion of progesterone. Sertoli and granulosa cells secrete inhibin, which acts, by a feed-back mechanism, on FSH secretion and to a lesser degree on that of LH by the pituitary. Furthermore, it directly inhibits multiplication of spermatogonias and diminishes progesterone secretion by the granulosa cells, as shown by in vitro studies. The LH uptake inhibitor (LH-RBI) is found in the corpora lutea in increasing quantities with age of this ovarian structure, and prevents luteal and granulosa cell progesterone secretion. This cybernine is probably one of the luteolytic factors. Gonadocrinins secreted by Sertoli and granulosa cells stimulate gonadotrophins liberation from pituitary cells such as hypothalamic LH-RH. It could be the same or slight by different factors. They act on specific receptors in Leydig's cells, the internal theca and the corpora lutea, leading to a reduction in LH receptors and thus the formation of sex steroids. The chemical structure of the cybernines is unknown, as well as their physiological and biochemical interrelationships. PMID- 6297371 TI - Regulation of expression of liver-specific enzymes. III. Further analysis of a series of rat hepatoma X human somatic cell hybrids. AB - 1. The expression of twelve liver-specific enzymes was analysed in twenty-one independent rat hepatoma X human somatic cell hybrids, and in some cases up to forty-one subclones were also tested. 2. Seventeen hybrids continued to express most of the rat liver-specific enzymes and in some cases human isozymes of glutamate-pyruvate transaminase, alpha-glycerophosphate dehydrogenase, guanine deaminase, alcohol dehydrogenase and pyruvate kinase were clearly identified. 3. Analysis of the segregation of the human liver-specific enzymes in these hybrids led to the assignment of human GPT to chromosome 8 (previously reported, Kielty, Povey & Hopkinson, 1982) and suggests the assignment of human GPD1 to chromosome 12. 4. The expression of the various liver-specific enzymes in these hybrids appeared to be controlled by independent regulatory mechanisms. 5. Four unusual reverse segregant hybrids were also analysed, and in these no liver-specific enzyme activity was demonstrable. PMID- 6297372 TI - Childhood brain tumors presenting as chronic uncontrolled focal seizure disorders. AB - Sixteen of 35 patients (46%) 21 years old or less who underwent surgical resection of a longstanding epileptic focus harbored tumors. The median duration of the seizure disorder prior to operation was 6.0 years for patients with and 7.6 years for those without tumor. Among the 35 patients, tumor was more common when intelligence and results of neurological examination were each normal, a plausible cause for uncontrolled seizures was lacking, and persistent focal delta activity occurred in a majority of electroencephalograms (EEGs). The type of seizures and the distribution of EEG spikes failed to distinguish patients with tumor from those without. Multifocal EEG spikes appeared in a majority of each group. PMID- 6297373 TI - DNA uptake in Haemophilus transformation. PMID- 6297374 TI - Interaction between host and viral genomes in mouse mammary tumors. PMID- 6297375 TI - Genetic defects in human purine and pyrimidine metabolism. PMID- 6297376 TI - T-DNA of the Agrobacterium Ti and Ri plasmids. AB - The study of T-DNA transmission from Agrobacterium Ti or Ri plasmid into the genomes of higher plant cells has revealed much about the consequences of transformation. It is now clear that the transformed phenotype is caused by hormonal changes produced directly or indirectly by T-DNA genes. The opine synthases are enzymes encoded in T-DNA that function in the plant cell. Our level of understanding of T-DNA-encoded functions is already sufficient to reveal clear and feasible ways to exploit T-DNA as a gene vector. What remains to challenge the crown gall investigator are many questions of fundamental importance: What is the mechanism of the seemingly illegitimate recombination between T-DNA and plant DNA, and is this process catalyzed by bacterial or host plant enzymes, or both? Do T-DNA genes encode enzymes that catalyze biosynthesis of auxin- and cytokinin active substances? What gene in T-DNA confers immunity to A. tumefaciens, and what is its mode of action? Does T-DNA insert into random or specific sites in the host plant genome? Did T-DNA derive from plant genetic information or has prokaryotic DNA arrived at functional eukaryotic gene structure by convergent evolution? Although there is keen interest in T-DNA as a vector for genetic engineering, it holds equal interest as a unique interface between the biology of prokaryotes and eukaryotes. PMID- 6297377 TI - Homologous pairing and strand exchange in genetic recombination. PMID- 6297378 TI - Genetic approaches to the analysis of microbial development. PMID- 6297379 TI - Developmental genetics of maize. PMID- 6297380 TI - Topical enviroxime against rhinovirus infection. AB - Enviroxime, an inhibitor of rhinovirus replication, was studied in a double blind, placebo-controlled trial with 99 volunteers. The efficacy of a nasal-spray formulation of enviroxime was tested as pretreatment or as postchallenge treatment for rhinovirus type 4 infection. In the regimens used, drug administration neither prevented infection nor reduced the frequency of specific colds. The mean concentration of enviroxime in nasal washes (12 h after a dose) differentiated two groups of responders. Those in whom the drug concentration exceeded 100 ng/ml had some benefits, although these were statistically insignificant. PMID- 6297381 TI - Chemical alterations in cell envelopes of polymyxin-resistant mutants of Pseudomonas aeruginosa grown in the absence or presence of polymyxin. AB - The polymyxin-resistant mutant strains H181 and H185 of Pseudomonas aeruginosa, after growth in the absence or presence of polymyxin, were compared with the polymyxin-sensitive H103 strain as to their cell envelope protein composition (determined by slab polyacrylamide gel electrophoresis) and their lipid composition. When grown in the absence of polymyxin, the H181 and H185 strains had an increased content of the outer membrane protein H1 with a decreased content of the outer membrane proteins D2 and F. After growth in the presence of polymyxin, the content of H1, D2, and F were all decreased. Significant alterations in the lipid composition of the H181 and H185 strains were found after growth in the absence of polymyxin. These lipid alterations were enhanced upon growth in the presence of polymyxin, with both strains having a reduced content of phosphatidylethanolamine and phosphatidylglycerol and an increased content of diphosphatidylglycerol and an unidentified lipid thought to be a neutral lipid. Other workers have proposed that an increased content of H1 protein is the molecular basis for polymyxin resistance in the H181 and H185 strains. Our observations make this hypothesis appear unlikely. PMID- 6297382 TI - Synergism among BIOLF-62, phosphonoformate, and other antiherpetic compounds. AB - 9-[[2-Hydroxy-1-(hydroxymethyl)ethoxy]methyl]guanine (BIOLF-62) is highly synergistic with either phosphonoformate or phosphonoacetate when used in combination against herpes simplex virus types 1 and 2 in vitro. Acycloguanosine did not show significant synergism with these two compounds. Bromovinyldeoxyuridine and phosphonoformate were highly synergistic against herpes simplex virus type 2, but not against type 1. PMID- 6297383 TI - Tromantadine: inhibitor of early and late events in herpes simplex virus replication. AB - Unlike amantadine (1-adamantanamine), tromantadine (N-1-adamantyl-N-[2-(dimethyl amino)ethoxy]acetamide hydrochloride) inhibits herpes simplex virus type 1 (KOS strain)-induced cytopathic effect and virus replication with limited toxicity to the cells. Vero and HEp-2 cells tolerated up to 2 mg of tromantadine per 2 X 10(6) cells for 24-, 48-, or 96-h incubation periods with little change in cell morphology. Treatment of the cells with 10 to 50 micrograms of tromantadine reduced herpes simplex virus-induced cytopathic effect. Treatment with 100 to 500 micrograms of tromantadine inhibited herpes simplex virus-induced cytopathic effect and reduced virus production. Complete inhibition of virus production was observed with treatments of 500 micrograms to 1 mg. The antiherpetic activity of tromantadine was dependent upon the viral inoculum size and the time of addition of the compound with respect to infection. Virion synthesis and viral polypeptide synthesis were inhibited by addition of tromantadine at the time of infection or 4 h postinfection. The results are consistent with tromantadine inhibition of an early event in herpes simplex virus infection, before macromolecular synthesis, and a late event, such as assembly or release of virus. PMID- 6297384 TI - Therapeutic activities of cefazolin, cefotaxime, and ceftazidime against experimentally induced Klebsiella pneumoniae pneumonia in rats. AB - The efficacies of several dosage schedules of cefazolin, cefotaxime, and ceftazidime, started 12 or 36 h after infection, were examined in experimental pneumonia caused by Klebsiella pneumoniae in rats. The therapeutic activities of the cephalosporins were compared with the antibacterial activities in vitro and the serum concentration curves. The course of experimental pneumonia was rapid and characterized by tissue necrosis. Response to antimicrobial treatment was evaluated with respect to mortality and numbers of bacteria in lung (left lobe), blood, and pleural fluid. When antibiotic treatment was started early, i.e., 12 h after bacterial inoculation, cefotaxime and ceftazidime were equally effective and superior to cefazolin. Eleven doses of 10 mg of cefotaxime or ceftazidime per kg or 11 doses of 60 mg of cefazolin per kg were required to improve the survival rate. With a delay in administration to 36 h after inoculation, the efficacy of the cephalosporins decreased markedly. In the three dosages tested, cefazolin was ineffective. Survival improved with the administration of nine doses of 60 mg of cefotaxime per kg or nine doses of 10 mg of ceftazidime per kg. These results are not in accordance with the ratio of in vitro activities of cefotaxime and ceftazidime or the serum concentration curves. PMID- 6297385 TI - Structure-activity relationships of new antiviral compounds. AB - In preliminary experiments, the compound 2-amino-5-(2-sulfamoylphenyl)-1,3,4 thiadiazole (G413) was shown to possess high activity against DNA viruses (herpes simplex viruses 1 and 2 and adenovirus 17) and RNA viruses (poliovirus 1, echovirus 2, and coxsackievirus B4). Experiments on the replicative cycle of poliovirus 1 and production of infectious RNA viruses demonstrate that this compound probably prevents assembly of virus particles by acting on structural proteins. In the present experiments, results concerning the activity of derivatives of G413 after side-chain modification are reported. Modification of the primary amine H to CH3 or CH2-CH = CH2 produced a loss of activity against DNA viruses, but inhibitory action on RNA viruses was preserved. Modification to CH2CH3 resulted in the loss of antiviral activity. PMID- 6297386 TI - Effect of protein synthesis inhibition on the induction of cell lysis in Escherichia coli by mecillinam plus nocardicin A. AB - Cell lysis can be induced in Escherichia coli by the combined use of the beta lactam antibiotics nocardicin A and mecillinam. Apparently, the induction of cell lysis occurs by a sequential mechanism in which mecillinam acts first, and nocardicin A has a triggering action insensitive to the inhibition of protein synthesis. PMID- 6297387 TI - Pharmacokinetics of cefotiam in normal humans. AB - Doses of 0.5, 1.0, and 2.0 g of cefotiam were infused intravenously over a 15-min period in a crossover fashion to eight volunteers. Doses of 1.0 and 2.0 g were infused intravenously over periods of 30 and 60 min in a double crossover fashion to another eight volunteers. Serum concentrations fell rapidly from peak levels between 30 and 170 micrograms/ml at the end of the infusion to less than 1.0 micrograms/ml within 6 h after all regimens. The terminal half-life in plasma varied between 0.6 and 1.1 h. The slopes of the time-concentration curves with the different regimens showed different half-life data. The urinary excretion of cefotiam was mostly completed within 4 h of the beginning of drug administration. The pharmacokinetics of cefotiam were dose dependent. With the 2-g dose, the peak plasma concentrations and the values for the area under the curve were more than twice the values observed with 1 g; decreasing values of plasma clearance were observed with higher doses. Injection of cefotiam caused no immediate discomfort or reaction at the infusion site. PMID- 6297388 TI - Inefficient accumulation of low levels of monodispersed and feces-associated poliovirus in oysters. AB - The accumulation of low levels (0.002 to 0.18 PFU/ml) of both feces-associated and monodispersed poliovirus by oysters (Crassostrea virginica or C. gigas) and clams (Mercenaria mercenaria) was investigated. These levels were chosen to duplicate the conditions present in light to moderately polluted waters. Experiments were performed in both small- and large-scale flowing seawater systems, developed to mimic the natural marine habitats of shellfish. Under these experimental conditions, viral accumulation by physiologically active shellfish was only noted when water column concentrations exceeded approximately 0.01 PFU/ml. Bioaccumulation increased with increasing concentrations of both monodispersed and feces-associated viruses. At virus concentrations below this level, viruses were seldom detected in either clams or oysters. Evidence indicated that the lack of accumulation was not the result of inefficient extraction or detection methods. The modified Cat-Floc-beef extract procedure used in the experiment was found to be capable of detecting as few as 1.5 to 2.0 PFU per shellfish. Evidence is presented to indicate that an uptake-depuration equilibrium was present at virus exposure levels of 0.10 PFU/ml, but not at 0.01 PFU/ml. The results suggested that viral accumulation by shellfish may not be efficient at water column concentrations below congruent to 0.01 PFU/ml. PMID- 6297389 TI - Viricidal capability of resin-triiodide demand-type disinfectant. AB - Polyoma, Newcastle disease virus, and adenovirus, as well as two coliphages, lambda and T4, were inactivated by strong base quaternary ammonium anion-exchange resin-triiodide. Organic matter interfered with viral inactivation capability of the resin-triiodide. The viruses, as they were being inactivated by the resin disinfectant beads, were not retained or filtered by the beads. PMID- 6297390 TI - Recovery of viruses from vegetable surfaces. AB - The efficiency of a system developed for the recovery of viruses contaminating large quantities of vegetables was investigated in the laboratory and tested in the field. Viruses seeded onto a number of leafy vegetables in the laboratory were eluted with a phosphate-buffered saline solution (pH 9.0). The eluate was clarified by glass wool filtration, and any viruses present were concentrated by adsorption to a Filterite pleated cartridge filter, eluted with 3% beef extract (pH 9.0), and further concentrated by organic flocculation. At least 24 liters of vegetable eluate could be concentrated to 70 to 80 ml, equivalent to a greater than 99.5% reduction in volume. With this system, poliovirus was recovered with a mean efficiency of 58% for all vegetables tested. Adenovirus was recovered from lettuce with a slightly lower mean efficiency (55%). Poliovirus was recovered from large quantities of cabbage for up to 5 days in the field after spray irrigation of relatively low levels of virus, even when heavy rain fell before sampling. PMID- 6297391 TI - Effect of chlorine treatment on infectivity of hepatitis A virus. AB - This study examined the effect of chlorine treatment on the infectivity of hepatitis A virus (HAV). Prodromal chimpanzee feces, shown to induce hepatitis in marmosets (Saguinus sp.), was clarified, and the virus was precipitated with 7% polyethylene glycol 6000, harvested, and resuspended. The suspension was layered onto 5 to 30% linear sucrose gradients and centrifuged; the fractions containing HAV were dialyzed, and a 1:500,000 dilution of this preparation induced hepatitis and seroconversion in 2 of 4 marmosets. A 1:50 dilution of this preparation served as inoculum. Untreated inoculum induced overt hepatitis and seroconversion in 100% (5 of 5) of marmosets inoculated intramuscularly. Inoculum treated for various periods (15, 30, or 60 min) with 0.5, 1.0, or 1.5 mg of free residual chlorine per liter induced hepatitis in 14% (2 of 14), 8% (1 of 12), and 10% (1 of 10) of marmosets, respectively, and induced seroconversion in 29, 33, and 10% of the animals. Inoculum treated with 2.0 or 2.5 mg of free residual chlorine per liter was not infectious in marmosets as determined by absence of hepatitis and seroconversion in the 13 animals tested. Thus, treatment levels of 0.5 to 1.5 mg of free residual chlorine per liter inactivated most but not all HAV in the preparation, whereas concentrations of 2.0 and 2.5 mg of free residual chlorine per liter destroyed the infectivity completely. These results suggest that HAV is somewhat more resistant to chlorine than are other enteroviruses. PMID- 6297392 TI - Accumulation of sediment-associated viruses in shellfish. AB - The present study focused on the importance of contaminated sediments in shellfish accumulation of human viruses. Epifaunal (Crassostrea virginica) and infaunal (Mercenaria mercenaria) shellfish, placed on or in cores, were exposed to either resuspended or undisturbed sediments containing bound poliovirus type 1 (LSc 2ab). Consistent bioaccumulation by oysters (four of five trials) was only noted when sediment-bound viruses occurred in the water column. Virus accumulation was observed in a single instance where sediments remained in an undisturbed state. While the incidence of bioaccumulation was higher with resuspended rather than undisturbed contaminated sediment, the actual concentration of accumulated viruses was not significantly different. The accumulation of viruses from oysters residing on uninoculated sediments. When clams were exposed to undisturbed, virus-contaminated sediments, two of five shellfish pools yielded viral isolates. Bioaccumulation of undisturbed sediments by these bivalves was considered marginal when related to the concentration of virus in contaminated sediments; they would only represent a significant threat when suspended in the water column. Arguments were advanced for water-column sampling in the region of the water-sediment interface to provide an accurate determination of the virological quality of shellfish harvesting waters. PMID- 6297393 TI - Effect of effluent from a nitrogen fertilizer factory and a pulp mill on the distribution and abundance of Aeromonas hydrophila in Albemarle Sound, North Carolina. AB - The density of Aeromonas hydrophila, standard count bacteria, fecal coliform bacteria, and 18 physical and chemical parameters were measured simultaneously at six sites for 12 months in Albemarle Sound, N.C. One site was above and two sites were below the discharge plume of a Kraft pulping process paper mill. The fourth site was above and the remaining two sites were below the discharge point of a nitrogen fertilizer factory. The impact of the pulp mill on water quality was acute, whereas that of the nitrogen fertilizer factory was chronic and much more subtle. Diffusion chamber studies indicated that A. hydrophila survival is increased by pulp mill effluent and decreased by nitrogen fertilizer factory effluent. From correlation and regression analysis, A. hydrophila was found to be directly affected by phytoplankton density and, thus, indirectly by concentrations of phosphate, nitrate, and total organic carbon. These two point sources are suspect as indirect causes of red-sore disease epizootics, a disease of fish caused by A. hydrophila. PMID- 6297394 TI - Enzymatic inactivation of residual gentamicin after membrane filtration. AB - RESIDUAL GENTAMICIN ON A MILLIPORE MEMBRANE WAS INACTIVATED BY MODIFICATION WITH TWO DIFFERENT ENZYMES: gentamicin adenylyltransferase and 3-N-acetyltransferase. Both modifications allowed the growth of susceptible strains of Escherichia coli and Bacillus subtilis. PMID- 6297395 TI - Simple apparatus for collecting estuarine sediments and suspended solids to detect solids-associated virus. AB - Laboratory trials of a new sampler for collection of estuarine sediment associated virus resulted in a recovery effectiveness averaging 30% for two enteroviruses and rotavirus SA11. A minimal recovery potential of 54% was calculated when losses caused by virus concentration procedure inadequacies were excluded. Both sediment-associated and suspended solids-associated viruses were collected with the sampler. Recoveries of 61 and 60% poliovirus and rotavirus, respectively, were obtained from salt water-suspended, solids-associated virus. The unique advantage of the sampler for selective collection of virus-associated top layers of sediment, plus collection over extensive areas, resulted in recovery of more virus than was obtained with a commonly used dredge-type sampler. PMID- 6297396 TI - Anesthetic-ion channel interactions: the effect of lidocaine on the stability and transport properties of the membrane-spanning domain of band 3. PMID- 6297397 TI - Comparison of cytochromes P-450 with high activity toward benzo[a]pyrene purified from liver microsomes of beta-naphthoflavone and 3-methylcholanthrene-pretreated rats. PMID- 6297398 TI - Kinetic differences between two interconvertible forms of fructose-1,6 bisphosphatase from Saccharomyces cerevisiae. PMID- 6297399 TI - Different behaviors of benznidazole as free radical generator with mammalian and Trypanosoma cruzi microsomal preparations. PMID- 6297400 TI - Studies on the UDP-sugar hydrolases from Escherichia coli and Salmonella typhimurium. PMID- 6297401 TI - On the mechanism of inhibition of fructose 1,6-bisphosphatase by fructose 2,6 bisphosphate. PMID- 6297402 TI - Induction of fructose 1,6-bisphosphatase and glucose 6-phosphatase in fetal mouse liver. AB - Fructose 1,6-bisphosphatase and glucose 6-phosphatase were induced in organ cultures of liver tissues from 15- and 19-day-old fetal mice, using a culture method that allowed the tissues to be maintained for 7 days in the absence of serum. In cultures from 15-day-old fetal liver, both enzyme activities increased significantly per milligram of DNA after a lag period of 1 to 3 days. In cultures from 19-day-old fetal liver only glucose 6-phosphatase increased in the absence of inducer. N6,O2'-Dibutyryladenosine 3',5'-monophosphate enhanced the rate of increase in fructose 1,6-bisphosphatase and glucose 6-phosphatase activities. The minimum effective concentration of the cyclic nucleotide was approximately 10(-6) M. Dexamethazone inhibited the increase in fructose 1,6-bisphosphatase during culture for 7 days. Glucose 6-phosphatase activity was enhanced by dexamethazone in cultures from 19-day-old fetal liver, but was without effect on glucose 6 phosphatase in cultures from 15-day-old fetal liver. The minimum inhibitory concentration of dexamethazone was less than 10(-8) M. The results suggest a complicated effect of the cyclic nucleotide on the two enzymes in fetal mouse liver as well as different mechanisms of action of dexamethazone on the induction of two enzymes. PMID- 6297403 TI - Comparison of the liver glycogen synthase from normal and streptozotocin-induced diabetic rats. AB - Glycogen synthase in the liver extracts of short-term (3 days) streptozotocin induced diabetic rats is poorly activated by the endogenous synthase phosphatase as well as phosphatase(s) from the liver extracts of normal animals. However, synthase in the liver extracts of diabetic rats is readily activated by the 35,000 Mr rabbit liver protein phosphatase (H. Brandt, F.L. Capulong, and E. Y. C. Lee (J. Biol. Chem. 250, 8038-8044 (1975)). The purified synthases from normal and diabetic animals respond differently after incubations with three different phosphatases. Both normal and diabetic synthase are activated by the 35,000 Mr protein phosphatase; however, the total activity of diabetic, but not the normal, synthase is significantly increased. Normal, but not the diabetic, synthase is activated by a synthase phosphatase from normal rats; this activation is accompanied by an increase in total synthase activity. Incubation of the diabetic synthase with calf intestinal alkaline phosphatase results in a reduction of the total synthase activity, whereas synthase activity of the normal is not significantly affected. The reduction in total activity of the diabetic synthase by treatment with alkaline phosphatase was prevented by prior dephosphorylation with 35,000 Mr rabbit liver protein phosphatase. In addition to their differences in responses to different phosphatases, the normal and diabetic synthases are also different in their molecular weights as determined by sucrose density gradient centrifugation (154,000 +/- 17,000 (n = 6) for the normal and 185,000 +/ 15,000 (n = 8) for the diabetic synthase) and their kinetic properties. PMID- 6297404 TI - cAMP-dependent protein kinases from the insect Ceratitis capitata. AB - Among the components of the two cyclic nucleotide system of Ceratitis capitata pharate adults, two cAMP-dependent protein kinase activities have been identified and purified through a sequence of chromatographic procedures. The properties of both protein kinases, A-1 and A-2, were studied and characterized in comparison with those of other sources. Protein kinase A-2 from Ceratitis capitata corresponds to type I from mammals mainly concerning about the dissociating effect of histones. Protein kinase A-2 exhibited a molecular weight of 39,000 in the presence of cAMP, whereas in the absence of the cyclic nucleotide two components of 80,000 and 159,000 were present and attributed to the forms RC and R2C2, respectively. Protein kinase activities A-1 and A-2 were markedly inhibited by increasing ionic strength whereas the activity (-cAMP/+cAMP) ratio for protein kinase A-2 increased versus NaCl concentration. Histones H1 and H2B were the best substrates for both A-1 and A-2 activities; the high mobility group of insect proteins (HMG) were also notably phosphorylated by A-2 preparation. Among the cyclic nucleotides assayed for the protein kinase activity A-2, cAMP induced a high activation at the lowest concentrations although high cAMP concentrations decreased the protein kinase activity, possibly through binding to the catalytic site. The protein kinase A-2 preparations exhibited a complex kinetics due to the presence of two forms with different affinity for ATP; these forms may be related to the aggregation properties of the enzyme. PMID- 6297405 TI - Biosynthesis of bacterial glycogen: activator specificity of the ADPglucose pyrophosphorylase of Rhodopseudomonads. AB - The adenosine diphosphate glucose pyrophosphorylases from Rhodopseudomonas acidophila, Rhodopseudomonas blastica, Rhodopseudomonas globiformis, and Rhodopseudomonas viridis were purified to the extent that their regulatory properties could be studied. With the exception of the R. viridis enzyme, all the enzymes could be activated by pyruvate or its analog, oxamate. The most effective activator for all the enzymes was fructose 6-P. However, the R. globiformis and R. viridis ADP glucose pyrophosphorylases can also be activated by fructose 1,6 P2. Thus a new activator specificity class was observed for the R. viridis enzyme while the R. acidophila and R. blastica enzymes exhibited the same activator specificity previously observed for Rhodopseudomonas capsulata ADPglucose pyrophosphorylase. The R. globiformis enzyme, activated by fructose 6-P, fructose 1,6-P2, and by pyruvate had a similar activator specificity previously seen for the Rhodopseudomonas sphaeroides and Rhodopseudomonas gelatinosa enzymes. For some enzymes, the presence of activator increased the apparent affinity for the substrates and MgCl2. The activator also modulated the sensitivity of the R. viridis and R. acidophila enzymes to Pi inhibition and the R. blastica enzyme to AMP inhibition. ADPglucose is the glucosyl donor for glycogen synthesis in these bacteria. Thus, regulation of glycogen synthesis in these microorganisms is probably regulated by the ratio of the activator concentration to inhibitor concentration. PMID- 6297406 TI - Phosphorylation of troponin and myosin light chain by cAMP-independent casein kinase-2 from rabbit skeletal muscle. AB - Casein kinase-2 from rabbit skeletal muscle was found to phosphorylate, in addition to glycogen synthase, troponin from skeletal muscle, and myosin light chain from smooth muscle. Troponin T and the 20,000 Mr myosin light chain are phosphorylated by casein kinase-2 at much greater rates than glycogen synthase. The V values for the phosphorylation of troponin and myosin light chain are nearly an order of magnitude greater than that of glycogen synthase; however, the Km values for these two substrates are greater than that for glycogen synthase. The kinase activities with the various protein substrates are stimulated approximately three- and fivefold by 5 mM spermidine and 3 mM spermine, respectively. Heparin is a potent inhibitor of the kinase when casein, glycogen synthase, or myosin light chain is the substrate. However, with troponin as substrate the kinase is relatively insensitive to inhibition by heparin. The amount of heparin required for 50% inhibition with troponin as substrate is at least 10 times greater than with casein as substrate. The phosphorylation of troponin by casein kinase-2 results in the incorporation of phosphate into two major tryptic peptides, which are different from those phosphorylated by casein kinase-1. The site in myosin light chain phosphorylated by casein kinase-2 is different from that phosphorylated by myosin light chain kinase. PMID- 6297407 TI - Evidence that forskolin activates turkey erythrocyte adenylate cyclase through a noncatalytic site. AB - The diterpene forskolin has been reported to activate adenylate cyclase in a manner consistent with an interaction at the catalytic unit. However, some of its actions are more consistent with an interaction at the coupling unit that links the hormone receptor to the adenylate cyclase activity. This report adds support to the latter possibility. Under conditions that lead to stimulation of adenylate cyclase in turkey erythrocyte membranes by GTP, forskolin also becomes more active. Additional evidence to support an influence of forskolin upon adenylate cyclase via the GTP-coupling protein N includes the following: (i) forskolin, at submaximal concentrations, leads to enhanced sensitivity and responsiveness of isoproterenol-dependent adenylate cyclase activity in turkey erythrocyte membranes; (ii) under specified conditions, the nucleotide GDP, an inhibitor of the stimulating nucleotide GTP and its analog, guanyl imidodiphosphate (Gpp(NH)p), also markedly inhibits the action of forskolin; (iii) both Gpp(NH)p and forskolin are associated with a decrease in agonist affinity for the beta adrenergic receptor. However, actions of forskolin in the turkey erythrocyte are not identical to those of GTP: (i) forskolin is never as potent as Gpp(NH)p in activating adenylate cyclase; (ii) the magnitude of synergism between isoproterenol and forskolin is not equal to that observed with isoproterenol and Gpp(NH)p; (iii) at high concentrations, forskolin inhibits antagonist binding to the beta-receptor. Forskolin appears to have several sites of action in the turkey erythrocyte membrane, including an influence upon the adenylate cyclase regulatory protein N. PMID- 6297408 TI - Eccrine adenocarcinoma. A clinicopathologic study of 35 cases. AB - A clinicopathologic study was made of 35 patients with primary eccrine adenocarcinoma diagnosed in the past 20 years from among 450,000 consecutive skin biopsy specimens. Histologically the following four distinct variants were identified: eccrine porocarcinoma (18 cases), syringoid eccrine carcinoma (12 cases), mucinous eccrine carcinoma (three cases), and clear cell eccrine carcinoma (two cases). Overall, eccrine adenocarcinomas are destructive lesions with a tendency to local recurrence. The syringoid histologic variant appears to be well differentiated and may have a benign clinical course; the lesion remained localized to the skin in our 12 cases. Regional lymphatic and distant metastasis, however, occurred in two patients with eccrine porocarcinoma. PMID- 6297409 TI - Crystalloid inclusions in endothelial cells of cellular and capillary hemangiomas. A possible sign of cellular immaturity. AB - The fine structure of crystalloid inclusions were found in the endothelial cells of capillary (strawberry) hemangiomas and cellular hemangiomas in nine infants. The inclusions measured from 0.5 to 2.0 microns in diameter and showed parallel lamellar bands with a regular periodicity of about 180 to 300 A. The presence of crystalloid inclusions and the absence of typical rod-shaped tubulated Weibel Palade bodies suggest the immaturity and embryonic character of the endothelium present in the blood vessels of these kinds of hemangiomas. PMID- 6297410 TI - Multiple sclerosis. PMID- 6297411 TI - Ketoconazole therapy and exfoliative erythroderma. PMID- 6297412 TI - Herpes simplex infection. PMID- 6297413 TI - Widespread foreign-body granulomas and elevated serum angiotensin-converting enzyme. AB - A patient had extensive foreign-body granulomatous inflammation of multiple skin sites and of the inguinal lymph nodes with splenomegaly, cutaneous anergy to common skin antigens, and peripheral blood eosinophilia. The patient had an elevated serum angiotensin-converting enzyme level. Histologically, the granulomas were of the foreign-body type with lymphocytes, histiocytes, eosinophils, and giant cells, some that contained doubly refractile crystalline material. Electron-probe x-ray microanalysis identified silicon, magnesium, iron, calcium, phosphorus, zinc, titanium, and chromium in the crystalline material. These findings suggest talc, cement, and inorganic pigment as possible sources of the crystals. This case is reported for its unusual clinical, laboratory, and morphologic features. PMID- 6297414 TI - Centennial paper. May 1912 (J Cutan Dis Syph 1912;30:241-255). Precancerous dermatoses: a study of two cases of chronic atypical epithelial proliferation. By John T. Bowen, M.D., Boston. PMID- 6297415 TI - Carcinoma of the proximal extrahepatic biliary tree radiologic assessment and therapeutic alternatives. AB - Preoperative cholangiography and angiography have been used in a series of 37 patients with hilar cholangiocarcinoma investigated over a 17-month period in a single specialist unit. Twelve lesions were judged to be irresectable on the basis of the cholangiographic findings: this was confirmed at laparotomy in nine patients and at autopsy in one. Angiography was performed in 21 patients, and suggested irresectability in eight: this was confirmed by laparotomy in seven. The eighth patient had compression of the left portal vein which had been interpreted as tumor invasion on angiography, and it was possible to perform a curative extended right hepatic lobectomy. Of the 13 potentially resectable patients, three were unfit for major resectional surgery. Five were found to be irresectable at laparotomy because of vena cava involvement in two and distant metastases in three. Five patients underwent resection with histologically clear margins. The combined use of cholangiography and angiography is recommended as a means of selecting appropriate therapy for patients with hilar cholangiocarcinoma. PMID- 6297416 TI - The value of needle renal allograft biopsy. I. A retrospective study of biopsies performed during putative rejection episodes. AB - Following renal transplantation, immunosuppression is usually increased to treat presumed rejection episodes. However, a) many conditions mimic rejection in the post-transplant period, and b) many rejection episodes are irreversible. As increased immunosuppressive therapy is associated with an increased risk of infection, it would be ideal to limit antirejection therapy to only the rejection episodes that are reversible. The role of percutaneous allograft biopsy was studied as an aid to decide which patients to treat for rejection, to limit unnecessary immunosuppression and to predict allograft survival. One hundred thirty-five patients with suspected rejection underwent 206 allograft biopsies without complication. Two hundred four biopsies were available for study. Biopsies were coded on a 1-4 scale (minimal, mild, moderate, severe) for acute and chronic tubulointerstitial infiltrate and vascular rejection, as well as no rejection (e.g., recurrence of original disease). Treatment decisions were made on the basis of the biopsy combined with clinical data. All patients have been followed two years and outcome correlated with biopsy findings (death, nephrectomy, and return to dialysis defined as kidney loss). The results were the following: 1) biopsies represented changes within the kidney. Of 16 kidneys removed within one month of biopsy, no nephrectomy specimen showed less rejection than that seen on biopsy. 2) Eighty-one biopsies (39.7%) led to tapering or not increasing immunosuppression (either no rejection, minimal rejection, or irreversible changes). 3) Kidneys having either severe acute or chronic vascular rejection (less than 30% function at three months) had significantly (p less than 0.05) decreased survival three to 24 months postbiopsy than those with minimal or mild vascular rejection or tubulointerstitial infiltrate (83% function at three months). 4) Kidneys with moderate chronic vascular rejection and those with severe acute tubulointerstitial infiltrate had significantly (p less than 0.05) decreased survival at six to 24 months. 5) Kidneys with moderate chronic vascular rejection (MCV) without an acute infiltrate (ATI) had significantly better survival than those having both MCV and ATI. 6) Similarly, kidneys having severe ATI alone had better survival than those with ATI plus vascular rejection. It was concluded that a) percutaneous allograft biopsy can be done without graft loss or infection; b) biopsy represents changes throughout the kidney; c) biopsy aids in deciding when to treat for rejection and in deciding when to withhold increased immunosuppression, and d) allograft biopsy predicts the outcome of treatment of a rejection episode. PMID- 6297417 TI - Abdominal wound closure. A randomized prospective study of 571 patients comparing continuous vs. interrupted suture techniques. AB - A randomized, prospective study was designed to compare a continuous with an interrupted technique for closing an abdominal incision. Five hundred seventy-one patients were randomized between the closure methods and stratified as to type of wound: clean, clean-contaminated, or contaminated. In mid-line incisions, the dehiscence rate was 2.0% (5/244) for the continuous group versus 0.9% (2/229) for the interrupted group. The difference was not statistically significant. Ventral hernias formed in 2.0% (4/201) of the continuous group vs. 0.5% (1/184) of the interrupted group. The type of wound had no influence on the results. In oblique incisions, 0% (0/39) of wounds closed continuously dehised while 2% (1/50) of incisions closed interruptedly dehised. No ventral hernias formed. Further analysis of the data indicated that dehiscence was more likely related to improper surgical technique than to the method of closure. An abdominal incision could be closed with a continuous suture in approximately half the time required for placing interrupted sutures (20 vs. 40 minutes). A continuous closure is preferred because it is more expedient and because it has the same incidence of wound disruption compared with an interrupted closure. PMID- 6297418 TI - Left ventricular function in patients with ventricular arrhythmias and aortic valve disease. AB - Forty patients having aortic valve replacement were evaluated preoperatively for ventricular arrhythmia and left ventricular ejection fraction. Arrhythmias were classified as complex or simple using the Lown criteria on the 24-hour ambulatory electrocardiogram; ejection fractions were determined by radionuclide gated blood pool analysis and contrast angiography. The ejection fractions determined by radionuclide angiography were 59.1 +/- 13.1% for 26 patients with simple or no ventricular arrhythmias, and 43.9 +/- 20.3% for 14 patients with complex ventricular arrhythmias (p less than 0.01). Ejection fractions determined by angiography, available for 31 patients, were also lower in patients with complex ventricular arrhythmias (61.1 +/- 16.3% versus 51.4 +/- 13.4%; p less than 0.05). Seven of 9 patients showing conduction abnormalities on the electrocardiogram had complex ventricular arrhythmias. Eight of 20 patients with aortic stenosis had complex ventricular arrhythmias, while 2 of 13 patients with aortic insufficiency had such arrhythmias. It is concluded that decreased left ventricular ejection fraction, intraventricular conduction abnormalities, and aortic stenosis are associated with an increased frequency of complex ventricular arrhythmias in patients with aortic valve disease. PMID- 6297419 TI - Perioperative myocardial infarction and changes in left ventricular performance related to coronary artery bypass graft surgery. AB - Recent advances in perioperative management and surgical technique have been associated with low operative mortality and a high incidence of symptomatic relief in patients undergoing coronary artery bypass graft (CABG) operations. The frequency and importance of perioperative myocardial infarction (MI) and immediate as well as long-term changes in left ventricular (LV) performance are factors of considerable current interest in any critical analysis of the effectiveness of CABG surgery. The present review describes the effects of patient selection, anesthetic techniques, and newer methods of myocardial protection as they relate to perioperative MI and LV performance. In addition, newer tests useful in the diagnosis of perioperative MI are discussed. The application of noninvasive techniques for the serial determination of LV performance and myocardial perfusion in CABG operations is also described. PMID- 6297420 TI - Powassan virus encephalitis resembling herpes simplex encephalitis. AB - A boy from New York traveling in Nova Scotia had olfactory hallucinations and other signs of temporal lobe involvement, leading to a diagnosis of herpes simplex encephalitis. The patient was treated with vidarabine and made a complete recovery. However, hemagglutination inhibition, complement fixation, and neutralization tests identified Powassan virus (POW) as the pathogen. Shortly before his trip to Nova Scotia, the patient had traveled in an area where POW encephalitis had occurred in humans (the eastern part of the state of New York), and he also came in contact with a known reservoir of POW infection (a groundhog) at home. PMID- 6297421 TI - Fatal disseminated cytomegalovirus infection in an apparently normal adult. AB - An ostensibly normal 43-year-old man had intermittent fever, erythema multiforme, and leukopenia that evolved into fulminant hepatic and bone marrow necrosis. The cause of his illness was compatible with cytomegalovirus (CMV) infection since there was a fourfold increase in immunofluorescent antibody titer. This report suggests that CMV can produce a rapidly progressive and overwhelming infection in an apparently healthy adult. PMID- 6297422 TI - [Mediators of inflammation. Their nature and their role]. PMID- 6297423 TI - Plasma immunoreactive beta-endorphin levels in depression. Effect of electroconvulsive therapy. AB - Immunoreactive (ir) plasma beta-endorphin level was assayed in ten symptomatic patients with a unipolar major depressive disorder and in 16 psychiatrically normal controls matched for age and sex. Plasma ir-beta-endorphin level in depressed patients was similar to that in controls. All depressed patients was similar to that in controls. All depressed patients had a transient, approximately threefold increase in ir-beta-endorphin after each use of electroconvulsive therapy (ECT). The increase of plasma ir-beta-endorphin level after ECT parallels the transient elevation of adrenocorticotropic hormone level reported by others and probably reflects a hypothalamic response to ECT. PMID- 6297424 TI - [Intensive combined chemo- and radiotherapy in small cell bronchial carcinoma]. AB - 117 patients with small cell bronchogenic carcinoma stage III-IV were treated, 82 patients with a combined chemotherapy and radiotherapy and 35 patients with combined chemotherapy only. All schedules investigated were successful. Long term remissions were obtained only in patients with a relatively located tumor, who achieved complete remission in consequence of treatment. Therapy results will be improved in future by perfecting the existing methods of treatment. PMID- 6297425 TI - Studies on potential antiviral compounds, XXII. Synthesis and in vitro antiviral activity of 1-(hydroxyalkyl)-1H-benzimidazoles. PMID- 6297426 TI - [Results of a clinico-experimental evaluation of suicide prevention programs]. AB - With regard to methodological shortcomings and heterogeneity of the outcome of experimental studies in the field of suicide prevention only tentative conclusions can be drawn from the results. Suicide prevention programs where contact to patients is actively established and maintained seem to be more effective in reducing the rate of further suicide attempts than passive strategies which leave it to the patients to initiate and continue a therapeutic relationship. This advantage emerges more clearly when patients are repeatedly or even continually motivated to utilize advice and treatment facilities. The intensity of outpatient aftercare services may also have a considerable influence on the effectiveness of suicide prevention. PMID- 6297427 TI - [Spinal and subcortical somatosensory evoked potentials after stimulation of the tibial nerve]. AB - Evoked potentials in response to unilateral stimulation of the posterior tibial nerve at the ankle were recorded above the spinous processes L5, L1, C2, and at Cz' in 30 normal subjects. The "cauda-potential" recorded above L5 consists of two small negative peaks with a mean latency of 18 and 22.5 ms respectively, whereas the "cord-potential" recorded above L1 exhibited a peak latency of 21.2 ms and on average a three-times larger amplitude than the first of the two "cauda potentials" (Fig. 1). Leads from the spinous process C2 revealed a sharp negative peak with a mean peak latency of 28.8 ms (N 30). Scalp recordings with a midfrontal (Fz-) reference inconsistently showed 1-2 small waves (P31, N33) prior to the primary cortical response (P40). Recordings with an ear- or non-cephalic reference consistently showed a large positive deflection (P30) which corresponded in latency with the simultaneously recorded cervical response (N30) and was followed by a distinct negative potential (N33) (Fig. 2a and b). Average latencies and amplitudes of the different spinal and subcortical evoked potentials (Tables 1 and 2), as well as the diagnostically more important interpeak-intervals, amplitude relations, and side-differences of latencies and amplitudes (Tables 3 and 4) were calculated. The diagnostic significance of these parameters will be shown in selected cases with spinal cord pathology. PMID- 6297428 TI - The proteins expressed by different isolates of Drosophila C virus. AB - Isolates of Drosophila C virus (DCV) from Drosophila flies obtained in geographically different regions were adapted to growth in Drosophila tissue culture cells. The viruses, purified from tissue culture cells, were shown to be serologically related to one of the isolates ("O" from Ouarzazate, Morocco). Analysis of the structural proteins by polyacrylamide gel electrophoresis demonstrated differences between the isolates. Labelling intracellular proteins of infected Drosophila melanogaster cells with 35S-methionine at 28 degrees C demonstrated the presence of the virus structural proteins and their immediate precursors. Raising the temperature to 37 degrees C both before and during the pulse period inhibited the processing of the high molecular weight proteins and resulted in a greater "shut-off" of host cell proteins than viral induced proteins. This allowed the precursor proteins to be compared as well as the structural proteins of the different strains. It was possible to clearly distinguish differences between the isolates on the basis of the induced proteins, although limited proteolysis of corresponding proteins showed marked similarities. Hence it is possible to distinguish between different isolates of the "same" small RNA-virus of insects from geographically different regions. PMID- 6297429 TI - Variation in cellular tropism between isolates of equine herpesvirus-1 in foals. AB - Subtype-1 isolates of Equine herpesvirus-1 (EHV-1) from a quadriplegic horse and from an aborted foetus were compared with each other and with a subtype-2 respiratory isolate. All 3 isolates were detected in the epithelium and macrophages of the respiratory tract. Both the paresis and foetal subtype-1 isolates replicated in the epithelium of the ileum and this correlated with the recovery of virus from faeces in vivo. The paresis subtype-1 isolate also had a predelection for vascular endothelial cells, particularly in the nasal mucosa, but also in the lungs, central nervous system, adrenal and thyroid. In the 9 foals inoculated with the paresis isolate two developed hind limb dysfunction, four developed diarrhoea, and one of these 4 died with an intussusception. The differences between these isolates are discussed in relation to other herpesviruses. PMID- 6297430 TI - Competition for cellular receptor sites among selected aphthoviruses. AB - The competition between different types of aphthoviruses (foot-and-mouth disease virus [FMDV]) for receptor site utilization was determined. The Southern African Territories (SAT) types of FMDV absorbed poorly to BHK-21 cells as measured by a radioactivity binding assay but grew to relatively high titers on these cells. On BK cells, however, all three SAT types bound well and competed with each other for receptor sites. In addition, unlabeled FMDV types A12 and O1B were able to completely inhibit the binding of 3H-uridine labeled SAT types. Unlabeled SAT, however, was only slightly able to inhibit the adsorption of labeled A12 and moderately inhibit the binding of labeled O1B. Saturation binding studies with homologous virus showed that BK cells contain at least 100-fold more receptor sites for types A12 and O1B than for the SAT types. Competitive binding analysis between type A12 FMDV and poliovirus and encephalomyocarditis virus revealed that these three viruses all used different receptor sites. Thus, different FMDV serotypes appear to utilize both common and unique receptor sites which are different from those of at least two other picornaviruses. PMID- 6297431 TI - Polymorphism of genomic RNAs within rotavirus serotypes and subgroups. AB - The genome of human rotaviruses is composed of 11 segments of double stranded RNA. Analysis by polyacrylamide gel electrophoresis of the genomic RNA from a number of strains of the same serotype has demonstrated a significant polymorphism in the migration pattern. However, two strains differing from each other in serotype were found to have apparently identical RNA migration patterns. It is concluded that serotype cannot be deduced from RNA migration patterns. Degrees of polymorphism have been observed within each of three known human rotavirus serotypes. PMID- 6297432 TI - Expression of virus-like particles in normal and transformed gerbil cells. AB - Three types of virus-like particles (VLP) were observed in three of seven gerbil cell lines. One of these cultures was in a non-transformed state and contained R and C-type VLP, while the two others began to express R-type or intracisternal A type VLP only after spontaneous transformation. These particles seem to be endogenous to the gerbil but do not appear to be directly involved in cell transformation. PMID- 6297433 TI - [Cancer of the mammary gland in a tigress]. AB - Mammary gland carcinoma in a 14-year-old female tiger kept in the Moscow Zoo for a long time is described. At autopsy, the outgrowth of the tumor into the surrounding tissue and metastases into the internal organs were found. The tumor was defined as a ductal papillary solid carcinoma with intraductal central necrosis that resembled the comedo type of human mammary gland carcinoma. Rapid growth of the tumor, the prevalence of necrosis, and numerous metastases to the lungs, liver, and lymph nodes were noted. PMID- 6297434 TI - [Method of preparation of the specimens for combined immunocytological examinations by light and electron microscopy]. AB - A method for making preparations for combined immunocytological examinations in light and electron microscopes is described. An infected cell culture grown on slides (a piglet kidney cell culture infected with Teschen virus) was fixed with 2.5% glutaraldehyde in phosphate buffer, pH 7.2-7.4 for 10-15 min and incubated with a conjugate (peroxidase-labeled immune serum to the causative agent of the infection under study). After postfixation, cytochemical reaction for peroxidase was performed, then the material was fixed with 2% OsO4 solution in phosphate buffer, pH 7,2-7,4, dehydrated in an ascending series of alcohols and gradually impregnated with polymerising resins (epon, araldit, etc.). The cells were embedded in foil baths, and after polymerization and separation of the slide and foil preparations with good light microscopy properties. In the light microscope, the desired parts of cells or a cell were selected, removed, cut in an ultratome and examined in electron microscope. PMID- 6297435 TI - [Neurologic manifestations of hyperparathyroidism. Study of 120 cases]. AB - The neurologic findings in 120 patients with possible diagnosis of hyperparathyroidism are reported and the world literature on this matter is reviewed. The neurologic signs and symptoms were divided in 3 categories: central, peripheral and miscellaneous. Headache, cramps weakness and depression where the most frequent complaints but with a so wide list of symptoms we expect mainly to salient the importance of calcium and phosphorus blood levels on neurologic patients as well as the value of neurologic evaluation on hyperparathyroid patients. PMID- 6297436 TI - Effects of synthetic muramyl dipeptide on bone metabolism in the fetal rat. AB - Synthetic N-acetyl-muramyl-L-alanyl-D-isoglutamine (MDP) stimulated the release of 45Ca from fetal rat long bones in culture for 48 h and also increased the number of osteoclasts per bone and the number of nuclei per osteoclast. It also increased calcium uptake in enriched populations of osteoclasts obtained by sequential collagenase digestions of newborn rat calvaria. This effect was observed after a 120 min incubation with 10(-6) M MDP. Effects of MDP on calcium uptake in osteoblasts were not consistently observed. No effects on cyclic AMP were noted in either osteoclast or osteoblast populations treated with MDP for periods from 1-120 min at 37 degrees C. PMID- 6297437 TI - Autonomic receptors in the developing submandibular glands of neonatal rats. PMID- 6297438 TI - Enzyme-assisted vitrectomy with bacterial collagenase. Time course and toxicity studies. AB - Electron microscopic analysis of intravitreal strands produced by the injection of autogenous fibroblasts showed thin, immature collagen after two weeks and the mature banded variety after four weeks. With the use of this intravitreal strand model, it was found that highly purified bacterial collagenase caused extensive digestion of scar tissue after incubation periods of 10, 15, and 30 minutes. There was no morphologic damage to cicatricial cellular elements or to the inner limiting membrane of the retina. A 45-minute exposure of retinas previously injured by photocoagulation to collagenase also did not result in morphologic evidence of damage. The use of collagenase as an adjunct to vitrectomy in cases of extreme vitreal scarring or retinal traction may decrease the complication rate of a procedure that is still extremely hazardous. PMID- 6297440 TI - Dual simultaneous systems for facial reanimation. AB - Successful reanimation of the paralyzed face requires a specific yet adaptable procedural armamentarium. Usually, in the treatment of regional paralysis, one distinct technique is deemed most appropriate and dependable. In cases of total hemiparesis secondary to surgical ablation or trauma, however, the simultaneous use of two separate but complementary rehabilitative systems has proved valuable in 15 patients. The reconstructive concept described divides the face into two functional spheres, an upper periorbital area and a lower perioral region. The integral system includes a direct facial nerve-to-cable graft reanastomosis for the upper division combined with a masseter muscle transfer for the lower facial region. The immediate supportive effects of the masseter transposition integrated with its long-term ability to rehabilitate via myoneurotization complement the more physiologically exacting effects of the nerve anastomosis. PMID- 6297439 TI - Fluorescein angiography of the heavily pigmented iris and new dyes for iris angiography. AB - An inexpensive adapter mounted in front of the objective lens of a fundus camera permits routine fluorescein angiography of the heavily pigmented iris. Angiograms of these patients whose iris pigment was dense enough that standard methods of iris angiography would disclose little or no information were performed with the use of the adapter. For use in the study of the iris vasculature in the research laboratory, three dyes with spectral characteristics different from those of fluorescein allow angiography of the iris at different wavelengths. Angiograms using these dyes were performed on the eye of a cynomolgus monkey. The angiograms were each taken as close as possible to the same time sequence and show dramatically different filling and staining patterns. PMID- 6297441 TI - Salivary gland imaging and radionuclide dacryocystography in agenesis of salivary glands. PMID- 6297442 TI - Foam cells in explants of atherosclerotic rabbit aortas have receptors for beta very low density lipoproteins and modified low density lipoproteins. AB - Beta-migrating very low density lipoproteins (beta--VLDL) and chemically modified low density lipoproteins (LDL), labeled with the fluorescent probe 3,3' dioctadecylindocarbocyanine (Dil), were used to determine whether foam cells from atherosclerotic lesions have lipoprotein receptors similar to those of macrophages. The advantage of the Dil probe is that it produces a brilliant fluorescence due to its retention by the cells. When cells grown from intimal medial explants of atherosclerotic rabbit aortas were incubated with Dil-labeled beta-VLDL, both smooth muscle cells and foam cells became fluorescent. However, when large amounts of unlabeled LDL were added to compete with the beta-VLDL for binding to the LDL receptors, Dil-labeled beta-VLDL binding to smooth muscle cells was blocked, and only the foam cells became highly fluorescent, due to the presence of specific receptors for beta-VLDL on those cells. The foam cells in the explants also became fluorescent when incubated with Dil-labeled acetoacetylated (AcAc) LDL. The same fluorescently labeled foam cells avidly bound IgG-coated erythrocytes. It was possible to inhibit the binding of the Dil labeled AcAc LDL to aortic foam cells by the addition of fucoidin, a known inhibitor of modified LDL binding to macrophages. Foam cells that became fluorescent were similar to cells grown from the explants that contained an abundance of Oil red O positive lipid droplets and demonstrated high acid lipase activity. These results suggest that macrophages were the precursors of the foam cells derived from explants of atherosclerotic rabbit aortas. One difference we noted between these foam cells and macrophages, however, was that foam cells from the explants retained their cholesteryl esters under the same culture conditions that resulted in the loss of lipid from macrophages loaded with lipoproteins in vitro. The existence of receptors for beta-VLDL on aortic foam cells supports the contention that dietary cholesterol-induced beta-VLDL may be atherogenic lipoproteins in vivo. PMID- 6297443 TI - Purification, morphology, and progesterone production and content of three cell types isolated from the corpus luteum of the sheep. AB - A method is presented for the isolation and purification of three cell types, endothelial cells, small luteal cells and large luteal cells, from the ovine corpus luteum. The method involves enzymatic dispersion of luteal tissue followed by centrifugation of separated cells on a Ficoll gradient. The three purified cell types and others, particularly fibrocytes and smooth muscle cells, that were removed during purification, were identified by their morphology. The cell yield, the cellular composition and cellular progesterone content of each fraction from the Ficoll gradient were measured. The endothelial cell fractions were relatively free of contamination by other cell types and had negligible progesterone. Fractions of small luteal cells and those of large luteal cells contained endothelial cells but were relatively free of other cell types. Large luteal cells contained significantly more progesterone, produced more progesterone when incubated in culture, but were less responsive to luteinizing hormone than small luteal cells. PMID- 6297444 TI - Differentiation of infectious laryngotracheitis virus strains using restriction endonucleases. AB - Strains of infectious laryngotracheitis virus (ILTV) were examined using an indirect immunofluorescent test (IIF) and with restriction endonucleases for detecting intratypic differences. Electrophoretic analysis of ILTV DNA fragments cleaved with restriction endonuclease Hind 111 clearly distinguished between strains. The IIF test did not discriminate between strains. A molecular weight estimate of ILTV DNA was made by summation of restriction endonuclease fragments cleaved with BamH1 (102.1 X 10(6)) and Hind111 (97.35 X 10(6)). Differences between the estimates may indicate the presence of submolar fragments. PMID- 6297445 TI - Influence of infectious bronchitis strains and vaccines on the incidence of Mycoplasma synoviae airsacculitis. AB - A model system was used to study infectious bronchitis virus (IBV) and Mycoplasma synoviae (MS) interaction. The system involved exposure of chickens to IBV, followed by exposure to MS 2-5 days later. The chickens were subjected to a cold environment (10 +/- 2 C) for 3 weeks starting one day before MS exposure. Under these conditions, differences in the capacity of various strains of IBV to exacerbate MS airsacculitis was demonstrated. Exposure to IBV field isolates generally resulted in more air-sac lesions than did higher-egg-passaged laboratory strains and vaccine strains. Use of lower-egg-passaged vaccines resulted in a higher incidence of airsacculitis than did higher-egg-passaged vaccines. When chickens were IBV-vaccinated before being used in the model system, the incidence of airsacculitis was lowered, even though the chickens became infected by the challenge virus. Vaccination of MS-free chickens with IBV had no effect on airsacculitis incidence when MS exposure occurred after the vaccine reaction was past. PMID- 6297447 TI - Propagation of virulent and avirulent turkey hemorrhagic enteritis virus in cell culture. AB - Virulent and apathogenic isolates of turkey hemorrhagic enteritis virus (HEV) were successfully propagated in lymphoblastoid cell lines of turkey origin, whereas spleen and kidney cell cultures from HEV-infected turkeys failed to replicate the virus. The lymphoblastoid cell lines used were MDTC-RP16 and MDTC RP19, which were previously established from tumors induced by Marek's disease virus in turkeys. Virus replication followed co-cultivation of lymphoblastoid cells with spleen cells from HEV-infected turkeys. Virus replication was demonstrated by immunofluorescence, by agar-gel-precipitin tests, and by electron microscopy. Supernatant fluid of cultures infected with virulent HEV caused death and specific lesions in turkey poults. Poults vaccinated with apathogenic HEV were protected against death and lesions after challenge with pathogenic HEV, which was recovered from infected cultures. The MDTC-RP19 cell line appeared far more susceptible than the MDTC-RP16 cell line to infection with HEV. PMID- 6297446 TI - Evaluation of ciliary movement in tracheal rings to assess immunity against infectious bronchitis virus. AB - Chickens vaccinated at one and 14 days of age with a live vaccine containing the Massachusetts and Connecticut strains of infectious bronchitis virus were protected against challenge with a Massachusetts strain administered at 5 weeks of age. The virus recovery method in which tracheal swabs taken 4 days post challenge were used to inoculate chicken embryos was compared with the method based on ciliary activity in tracheal explants. The results showed that evaluation of ciliary activity effectively measured immunity to infectious bronchitis virus. Serological results obtained in a microneutralization test using chicken embryo kidney cells correlated with the immune status of the birds as well as with the alpha neutralization results obtained using chicken embryos. PMID- 6297448 TI - Comparison of ciliary activity and virus recovery from tracheas of chickens and humoral immunity after inoculation with serotypes of avian infectious bronchitis virus. AB - Cross-protection tests with homologous and heterologous serotypes of infectious bronchitis virus (IBV) were used to compare ciliary activity and virus recovery from tracheas of chickens. Validation of this technique included correlating the neutralization indices of antiserum obtained from some infected birds. Chickens were inoculated intratracheally with either the JMK or Connecticut (Conn) serotype of IBV. Three weeks later, infected and uninfected groups were challenged by the same route with homologous and heterologous virus. The JMK strain provided immunity against homologous challenge and the Conn strain, as indicated by good ciliary activity and lack of challenge virus recovery. The Conn strain provided only homologous protection, as ciliostasis occurred and virus was recovered after challenge with the JMK strain. In each case, antiserum to immunizing virus neutralized only the homologous virus. Controls were uniformly susceptible and lacked neutralizing antibody. A similar experiment with the Ark 99 serotype and a recent isolate (397) of IBV revealed complete cross-protection of the tracheas. Antiserum to each virus neutralized the homologous and heterologous virus in each case in reciprocal tests. The results indicate that these two viruses are closely related. The complete agreement between ciliary activity and virus isolation indicates that ciliary activity is a reliable, objective criterion upon which tracheal immunity can be judged in cross protection tests. PMID- 6297449 TI - Field test studies of the 2512 strain of infectious bursal disease. AB - Responses to an infectious bursal disease vaccine made from the 2512 strain were studied in broiler flocks under field conditions and management. Pre- and post vaccination sera were analyzed by agar-gel-precipitin tests and beta virus neutralization. Specific measurements of broiler performance (livability, weight, feed conversion, and condemnations) were also analyzed. When possible, contemporary licensed products were compared at the same time in adjacent houses. Results of these trials indicate that the vaccine made from the 2512 strain of infectious bursal disease was highly immunogenic and safe under field conditions. PMID- 6297450 TI - Influence of maternal antibody on efficacy of embryo vaccination with cell associated and cell-free Marek's disease vaccine. AB - Vaccination with turkey herpesvirus (HVT) of 18-day-old chicken embryos from a commercial source or from a cross (15 X 7) of two inbred lines induced better protection against early post-hatch challenge with virulent Marek's disease virus (MDV) than vaccination at hatch, despite the presence in embryos of maternally derived antibodies to HVT or to HVT and MDV. However, 50%-protective-dose (PD50) assays revealed that maternal antibodies in embryos reduced vaccine efficacy. The PD50 assays were conducted by vaccinating 15 X 7 embryos with serial dilutions of HVT at the 18th day of incubation. Embryonally vaccinated and unvaccinated chicks were challenged with MDV on the day of hatch. In the absence of maternal antibodies, the PD50 values in plaque-forming units for cell-associated and cell free HVT were 57 and 328, respectively. In the presence of maternal antibodies, PD50 values for cell-associated and cell-free HVT were 105 and greater than 4,000, respectively. PMID- 6297451 TI - Characteristics and serologic studies of two serotypes of infectious bursal disease virus in turkeys. AB - Two serotypes of infectious bursal disease virus (IBDV) were distinguished using the virus-neutralization test. An IBDV isolate from chickens, two commercial IBDV vaccine viruses, and an IBDV isolate from turkeys were designated serotype I. Two IBDV isolates from turkeys were designated serotype II. A common antigen shared by serotype I and serotype II viruses was detected using an indirect immunofluorescent assay. The virions of both IBDV serotypes were determined to be nonenveloped icosahedrons with diameters ranging from 58 to 60 nm. The virion densities of one serotype I isolate and two serotype II isolates were similar (1.32 g/ml). Twenty-three turkey breeder flocks and 22 commercial turkey flocks from Ohio, North Carolina, and Indiana were tested for antibodies to the two IBDV serotypes. Seventy percent of the 23 breeder flocks had antibody titers to serotype I, and 75% had antibody titers to serotype II. None of the 22 commercial flocks had antibodies to serotype I, and 77% had antibody titers to serotype II. Antibodies to serotype I were detected only in breeder flocks that were vaccinated using a commercial serotype I IBDV vaccine. Only antibodies to serotype II viruses were naturally widespread in the flocks tested. PMID- 6297452 TI - Serological cross-reactivity of avian adenovirus serotypes in an enzyme-linked immunosorbent assay. AB - Fowl adenoviruses free of avian adenovirus-associated virus, representing 10 serotypes, were tested for cross-reactivity in an enzyme-linked immunosorbent assay (ELISA). All antigens and antisera prepared in chickens, along with uninfected control antigen and normal chicken serum, were reacted in a checkerboard pattern with ELISA. There was considerable cross-reactivity among all serotypes tested. Homologous reactions were generally, but not always, stronger than heterologous reactions. ELISA was about as sensitive as virus neutralization in detecting antibodies. The high sensitivity plus broad-spectrum reactivity should make ELISA a preferred test for the detection of adenovirus antibodies in poultry flocks. PMID- 6297453 TI - Marek's disease virus (MDV) antigens in the feather follicle epithelium: difference between oncogenic and nononcogenic MDV. AB - Feather tips from chickens infected with oncogenic or nononcogenic Marek's disease virus (MDV) were individually tested for MDV antigens by the agar-gel precipitin test to compare antigen contents of feather follicles infected with oncogenic or nononcogenic MDV. There was a significant difference between oncogenic and nononcogenic MDV in the development of MDV antigens in the feather follicle epithelium of infected chickens. Significantly more feather tips from chickens infected with oncogenic MDV were positive for MDV antigens, particularly at 3 and 6 weeks postinoculation, whereas the feather tips from those infected with nononcogenic MDV were rarely positive, whether tested with the homologous or heterologous anti-MDV serum. Such a difference was consistent in separate trials employing three different strains of chickens. Infectious cell-free MDV was extracted more frequently from the feather tips of chickens infected with oncogenic MDV than from those of birds infected with nononcogenic MDV, and the virus titers of extracted MDV were always numerically or statistically higher in birds infected with oncogenic MDV. PMID- 6297454 TI - Cardiac alpha- and beta-adrenergic receptor alterations in diabetic cardiomyopathy. AB - The effect of chronic experimental diabetes on the adrenergic receptors in the rat heart was investigated. Diabetes was induced by streptozotocin (65 mg/kg; i.v.) administration, animals were sacrificed 8 weeks later, and positive as well as negative dF/dt values were determined in isolated papillary muscle preparations. Stimulation of the contractile force generation by isoproterenol and methoxamine was attenuated in diabetic preparations. Beta- and alpha adrenergic receptor bindings were determined in cardiac membranes by employing 3H dihydroalprenolol and 3H-dihydroergocryptine respectively. Reduced number of beta and alpha-receptor binding sites without changes in the affinity constants were observed in diabetic myocardium. Such a decrease in alpha- and beta-receptor density in the heart may account for the depressed contractile responsiveness to adrenergic stimuli in diabetic cardiomyopathy. PMID- 6297455 TI - Mechanistic studies on the dehydrogenases of methylotrophic bacteria. 1. The influence of substrate binding to reduced trimethylamine dehydrogenase on the intramolecular electron transfer between its prosthetic groups. AB - The trimethylamine dehydrogenase of bacterium W3A1 is reduced with the formation of a triplet state in which two electrons, derived from the substrate, are distributed between the [4Fe-4S] cluster and 6-S-cysteinyl-FMN semiquinone. In titration experiments at pH 8.5 about 1.0 mol of dimethylamine or 0.5 mol of trimethylamine per mol of the enzyme is required to titrate the enzyme to an endpoint. At pH values less than 8.0, however, an excess of trimethylamine is required to obtain maximal yield of the g = 4 e.p.r. signal, characteristic of the triplet state, or maximal absorbance at 365 nm which indicates formation of the flavin semiquinone. The binding of 0.86 mol of trimethylamine per mol of the enzyme could be detected by a gel chromatographic method. When the enzyme is titrated with dithionite in the presence of tetramethylammonium chloride, an endpoint is reached after the uptake of two electrons which give rise to the triplet state, whereas three electrons are consumed in the absence of tetramethylammonium chloride to reduce the enzyme completely. The enzyme is inhibited noncompetitively by tetramethylammonium chloride and the slopes of double reciprocal plots are a concave upwards function of inhibitor concentration. The data indicate the presence of a binding site for the substrate and other amines on the reduced enzyme which enhances the proportion of enzyme in the triplet state. PMID- 6297456 TI - Mechanistic studies on the dehydrogenases of methylotrophic bacteria. 2. Kinetic studies on the intramolecular electron transfer in trimethylamine and dimethylamine dehydrogenase. AB - E.p.r. spectroscopy of the trimethylamine and dimethylamine dehydrogenases of Hyphomicrobium X indicates that the substrate-reduced forms of these enzymes exist in the triplet state, which arise through interaction of a reduced [4Fe-4S] cluster and flavosemiquinone, with e.p.r. signals which differ in detail from those of the trimethylamine dehydrogenase of bacterium W3A1. Under certain conditions the intramolecular electron transfer between the flavoquinol form of 6 S-cysteinyl-FMN and the [4Fe-4S] cluster in all three dehydrogenases was much slower than the preceding reduction of the flavin to the flavoquinol form. Trimethylamine dehydrogenases from both organisms show a time-dependent broadening of the e.p.r. signals centred around g = 2 after mixing with trimethylamine. The broadening of the e.p.r. signals could be correlated with an unexpected dependence of the rate of formation of the triplet state on substrate concentration. A model which accounts in a qualitative manner for the substrate dependence of the formation of the triplet state in the trimethylamine dehydrogenase of Hyphomicrobium X is proposed. The binding of the substrate to the reduced form of the enzyme seems to result in a conformational change of the enzyme to a form in which the rate of intramolecular electron transfer is decreased. This finding may be correlated with the observation of hyperbolic substrate inhibition for both trimethylamine dehydrogenases. The results indicate the transfer of an electron to the [4Fe-4S] cluster to be an obligatory step in catalysis and suggest that the transfer of electrons from these enzymes to electron acceptors is mediated solely through the [4Fe-4S] cluster. PMID- 6297457 TI - The inhibition of bovine and rat parotid deoxyribonuclease I by skeletal muscle actin. A biochemical and immunocytochemical study. AB - Rat and bovine parotid gland and pancreas contain deoxyribonuclease I (DNAase I) activities in different amounts. The DNAase I activity in tissue homogenates of bovine and rat parotid gland can be inhibited by addition of monomeric actin, as with the enzyme of bovine pancreas. The isolated DNAase I species from bovine and rat parotid gland differ in their molecular weights and also in their affinities for monomeric actin, being lowest for rat parotid DNAase I (5 X 10(6)M(-1). Antibodies raised against rat and bovine parotid and bovine pancreatic DNAase I can be used to study the subcellular localization of DNAase I in these tissues by indirect immunofluorescence. DNAase I was found to be confined solely to the secretory granules of the tissue from which it was isolated. PMID- 6297458 TI - Electron-spin-resonance studies of the NADH-dependent nitrite reductase from Escherichia coli K12. AB - The NADH-dependent nitrite reductase of Escherichia coli, which contains sirohaem, flavin, non-haem iron and labile sulphide, was examined by low temperature e.s.r. spectroscopy. The enzyme, stored in the presence of nitrite and ascorbate, gave the spectrum of a nitrosyl derivative, with hyperfine splitting due to the nitrosyl nitrogen. On removal of these reagents, a series of signals centred around g = 6 was observed, typical of high-spin ferric haem. Cyanide converted this into a low-spin form. On reduction of the enzyme with NADH, an axial spectrum at g = 1.92, 2.01 was observed. The temperature dependence of this signal is indicative of a [2Fe-2S] iron-sulphur cluster. The midpoint potential of this cluster was estimated to be -230 +/- 15 mV by two independent methods. Reduction of the enzyme with dithionite yielded further signals, which are at present unidentified, at g = 2.1-2.28. No signals were observed that could be assigned to a [4Fe-4S] cluster, such as is found in other sulphite reductases and nitrite reductases that contain sirohaem. PMID- 6297459 TI - Effects of colchicine and vinblastine on output of proteins into bile. AB - The microtubule poisons colchicine and vinblastine caused a reduction in the biliary output of total protein, compared with controls, in bile-fistula rats. The various protein components of bile showed patterns of change in output differing from one another. Alkaline phosphodiesterase I, a 'plasma-membrane' enzyme, showed a decline in output during the first hour after drug administration. Immunoglobulin A output did not decline until after the first hour. In contrast with these reductions, the biliary output of (rat) plasma albumin into bile was increased. At no time was there any evidence (as measured by lactate dehydrogenase release into bile) for any cytolytic damage. These results are discussed in relation to current theories on the output of proteins into bile and the repair of the plasma membrane in the bile canaliculus. PMID- 6297460 TI - Transferrin receptors of human fibroblasts. Analysis of receptor properties and regulation. AB - Normal human skin fibroblasts cultured in vitro exhibit specific binding sites for 125I-labelled transferrin. Kinetic studies revealed a rate constant for association (Kon) at 37 degrees C of 1.03 X 10(7) M-1 X min-1. The rate constant for dissociation (Koff) at 37 degrees C was 7.9 X 10(-2) X min-1. The dissociation constant (KD) was 5.1 X 10(-9) M as determined by Scatchard analysis of binding and analysis of rate constants. Fibroblasts were capable of binding 3.9 X 10(5) molecules of transferrin per cell. Binding of 125I-labelled diferric transferrin to cells was inhibited equally by either apo-transferrin or diferric transferrin, but no inhibition was evident with apo-lactoferrin, iron-saturated lactoferrin, or albumin. Preincubation of cells with saturating levels of diferric transferrin or apo-transferrin produced no significant change in receptor number or affinity. Preincubation of cells with ferric ammonium citrate caused a time- and dose-dependent decrease in transferrin binding. After preincubation with ferric ammonium citrate for 72 h, diferric transferrin binding was 37.7% of control, but no change in receptor affinity was apparent by Scatchard analysis. These results suggest that fibroblast transferrin receptor number is modulated by intracellular iron content and not by ligand-receptor binding. PMID- 6297461 TI - Underestimation of metabolic rates owing to reincorporation of 14CO2 in the perfused rat liver. AB - (14)CO(2) production by perfused rat livers was simulated by infusing NaH(14)CO(3) into the perfusate. Recovery of label as (14)CO(2) gas + perfusate bicarbonate was 45-85%. Rates of (14)CO(2) exchange in the liver are 3-70 times greater than net rates of CO(2) production. Therefore (14)CO(2) reincorporation can lead to significant underestimations of rates of oxidation of (14)C-labelled substrates in liver. PMID- 6297462 TI - Modulation of dexamethasone receptor expression in embryonal carcinoma cells and their differentiated derivatives. AB - The dexamethasone binding capacity of embryonal carcinoma cells and their differentiated derivatives was investigated. Manipulation of the embryonal carcinoma cell-culture conditions resulted in an unstable reversible expression of the glucocorticoid receptors. Stable expression of the receptors is observed when these cells are induced to differentiate. Cells grown under identical conditions were assayed for their ability to bind epidermal growth factor. PMID- 6297463 TI - Ascorbate suppresses the opiate-induced compensatory increase in cyclic AMP in neuroblastoma X glioma hybrid cells. AB - In NG108-15 hybrid cells ascorbate suppresses the delayed etorphine-induced compensatory increase in the levels of cyclic AMP. It has, however, no effect on the early response of the cells to etorphine, as manifested in a transient decrease in the levels of cyclic AMP. PMID- 6297464 TI - Purification of a protein having pore forming activity from the rat liver mitochondrial outer membrane. AB - A protein with pore-forming activity has been isolated from the outer membrane of rat liver mitochondria. The purification involves sucrose gradient centrifugation, differential centrifugation in the presence of Triton X-100, and DEAE-Sepharose and CM-Sepharose chromatography. The yield of the purified protein was approx. 2% of the total outer membrane proteins. The protein, when inserted into soya bean phospholipid vesicles, increases the [3H]sucrose permeability of the vesicles but had no effect on the permeability of high-molecular-weight [14C]dextran (Mr 70 000). The protein is very active, since as little as 3-4 micrograms of protein per mg of phospholipid is required for the complete release of [3H]sucrose from the vesicles. Sucrose diffusion channels could not be reconstituted with other membrane proteins such as rat liver cytochrome oxidase or cytochrome b5. Purified pore protein revealed a single band of apparent Mr 30000 when resolved by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis. This polypeptide could be further resolved by isoelectric focusing into a major (pI7.9) and two relatively minor (pI7.6 and 7.2) components. Proteolytic mapping with V8 proteinase from Staphylococcus aureus suggests that these probably represent a single component showing charge heterogeneity. The reason for the charge heterogeneity is not known. The amino acid composition of the protein revealed 47.8% polar amino acids with a relatively high lysine content. PMID- 6297465 TI - Influence of the state of ribosome association on the phosphorylation of ribosomal proteins in isolated ribosome--protein kinase systems from rat cerebral cortex. AB - Ribosomal protein phosphorylation was investigated in isolated ribosomal subunits and polyribosomes from rat cerebral cortex in the presence of [gamma-32P]ATP and purified catalytic subunit of cyclic AMP-dependent protein kinase from the same tissue. Ribosomal proteins that were most readily phosphorylated in isolated cerebral ribosomal subunits included proteins S2, S3a, S6 and S10 of the 40 S subunit and proteins L6, L13, L14, L19 and L29 of the 60 S subunit. These proteins were also phosphorylated in cellular preparations of rat cerebral cortex in situ or in vitro [Roberts & Ashby (1978) J. Biol. Chem. 253, 288-296; Roberts & Morelos (1979) Biochem. J. 184, 233-244]. However, several additional ribosomal proteins were phosphorylated when isolated 40 S or 60 S subunits were separately incubated in the reconstituted system. Analogous results were obtained with an equimolar mixture of cerebral 40 S and 60 S subunits under comparable conditions. In contrast, extensive exposure of purified cerebral polyribosomes to the catalytic subunit resulted in phosphorylation of only those ribosomal proteins of the 40 S subunit that were most highly labelled after the administration of [32P]Pi in vivo: proteins S2, S6 and S10. Ribosomal proteins of 60 S subunits that were readily phosphorylated in isolated cerebral polyribosomes included proteins L6, L13 and L29. These results indicate that polyribosome formation markedly decreases the number of ribosomal protein sites available for phosphorylation by the catalytic subunit of cyclic AMP-dependent protein kinase. Moreover, the findings suggest that, of the ribosomal protein phosphorylations observed in rat cerebral cortex in vivo, proteins S2, S6, S10, L6, L13 and L29 can be phosphorylated in polyribosomes, whereas proteins S3a, S5, L14 and L19 may become phosphorylated only in free ribosomal subunits. PMID- 6297466 TI - The hormone-binding unit of luteinizing hormone receptor. AB - Rat ovarian luteinizing hormone/human choriogonadotropin binding sites were labelled with 125I-choriogonadotropin in vivo, and the resulting 125I choriogonadotropin-receptor complexes were solubilized by Triton X-100 and purified by use of antibodies to choriogonadotropin immobilized to agarose. The purified 125I-choriogonadotropin-receptor complex was treated with glutaraldehyde to crosslink radiolabelled hormone to the receptor. Sodium dodecyl sulphate/polyacrylamide-gel electrophoresis of the crosslinked product revealed a labelled Mr 130 000 major band in addition to the hormone and its alpha-subunit, indicating that a single receptor component was linked to the hormone. Unoccupied binding sites for luteinizing hormone were also solubilized by Triton X-100 from pseudopregnant rat ovaries, and attached to choriogonadotropin-agarose. The agarose gel was washed, and eluted with 0.1 M-sodium acetate, pH 4. Sodium dodecyl sulphate/polyacrylamide-gel electrophoresis of the pH 4 eluate revealed an Mr 90 000 major band which was abolished when ovaries presaturated with choriogonadotropin were used as starting material. These observations suggest that the hormone-binding component of the luteinizing hormone receptor is a polypeptide of Mr 90 000. This polypeptide was isolated and labelled with Na 125I. The labelled polypeptide showed a single band on sucrose density gradient centrifugation and on gel filtration on agarose. PMID- 6297467 TI - Characteristics of the desensitization and resensitization of the cyclic AMP independent glycogenolytic response in rat liver cells. AB - Vasopressin and alpha-adrenergic agonists were shown previously [Breant, Keppens & De Wulf (1981) Biochem. J. 200, 509-514] to induce a heterologous, dose dependent and receptor-mediated desensitization of the cyclic AMP-independent glycogenolytic response in isolated hepatocytes. The desensitized state of the hepatocytes can be preserved as long as the agonist is bound to its receptor. Conversely, washing the cells with a hormone-free buffer or displacement of the agonist from its receptor by a specific antagonist restores the responsiveness. The desensitization and its reversibility (i.e. resensitization) are obtained within minutes. The desensitization can be clearly elicited at temperatures as low as 5 degrees C, whereas the glycogenolytic response and the enhancement of the 45Ca flux are only obtained above 15 degrees C; the resensitization requires even higher temperatures. A tentative model is proposed to account for the observed effects. PMID- 6297468 TI - Variable H+/substrate stoicheiometries in Rhodotorula gracilis are caused by a pH dependent protonation of the carrier(s). AB - Two carrier-mediated systems transport sugars in the yeast Rhodotorula gracilis depending on the pH. One system, with higher affinity for sugars, catalyses a symport of protons with sugar, whereas the other system, having lower affinity, is independent of protons. This was shown in three different ways. (1) At low pH, where only the high-affinity system works, a H+/sugar stoicheiometry of 1 was found. An increase of the pH and of the sugar concentration, which allowed the low-affinity system to operate, brought about a drop of the stoicheiometry to values below 1. (2) During H+ symport the influx of positive charge was electrically compensated by an equivalent efflux of K+ from the cells. At high pH and high sugar concentrations this stoicheiometry of K+ and sugar decreased concomitant with the H+/sugar stoicheiometry. (3) At pH 7.5 both transport systems were operating, as shown by biphasic saturation kinetics. Under these conditions only the high-affinity transport was found to be electrogenic. These results agree with the theory of an electrogenic H+/sugar symport where changes in the affinity for substrate are brought about by reversible protonation and deprotonation of the carrier. PMID- 6297469 TI - Cytochrome c oxidase is preferentially synthesized in the rough endoplasmic reticulum--mitochondrion complex in rat liver. AB - The specific activity and content of cytochrome oxidase in the rough endoplasmic reticulum--mitochondrion complex are higher than in the mitochondrial fraction. Radiolabelling studies with the use of hepatocytes and isolated microsomal and rough endoplasmic reticulum--mitochondrion fractions, followed by immunoprecipitation with anti-(cytochrome oxidase) antibody, reveal that the nuclear-coded cytoplasmic subunits of cytochrome oxidase are preferentially synthesized in the latter fraction. The results have a bearing on the mechanism of transport of these subunits into mitochondria. PMID- 6297470 TI - The control of glucose 1,6-bisphosphatase by Ca2+ and calmodulin. AB - 1. The control of glucose 1,6-bisphosphatase activity, the enzyme that degrades glucose 1,6-bisphosphate, a metabolite that regulates hexose phosphate metabolism, has been examined in a rat muscle extract. 2. The enzyme has been found to be activated by physiological Ca2+ concentrations. Ca2+ can be replaced by Sr2+ and Mn2+, but the effects are not so pronounced. 3. The Ca2+ effect is inhibited by EGTA. Trifluoperazine also inhibits enzyme activity. This effect can be reversed by adding exogenous calmodulin. PMID- 6297471 TI - Furosemide-sensitive K+ channel in glioma cells but not neuroblastoma cells in culture. PMID- 6297472 TI - Formation of leukotrienes and other hydroxy acids during platelet-neutrophil interactions in vitro. PMID- 6297473 TI - Sialic acid residues of the alpha-subunit are required for the thyrotropic activity of HCG. PMID- 6297474 TI - Phytohaemagglutin-induced changes in spin label reduction in lymphocytes from tumor-bearing rats. PMID- 6297475 TI - Hydroxyurea inhibits degradation of internalized epidermal growth factor in HeLa cells. PMID- 6297476 TI - The size of the mammalian lung beta 2-adrenergic receptor as determined by target size analysis and immunoaffinity chromatography. PMID- 6297477 TI - Degradation of phosphatidylinositol-4,5-bisphosphate is insensitive to CA2+ mobilization in stimulated platelets. PMID- 6297478 TI - A fluorometric-HPLC assay for quantitating the binding of benzo[a]pyrene metabolites to DNA. PMID- 6297479 TI - Recombinant JC viral DNA: verification and physical map of prototype. PMID- 6297480 TI - Phosphorylation of calf uterus 17 beta-estradiol receptor by endogenous Ca2+ stimulated kinase activating the hormone binding of the receptor. PMID- 6297481 TI - Tumor cell stimulation of collagenase production by fibroblasts. PMID- 6297482 TI - Rat hepatocyte proliferation is stimulated by insulin-like peptides in defined medium. PMID- 6297483 TI - Variation in the inhibition of restriction enzyme cleavage of lambda phage DNA produced by two covalent binding antitumor agents: anthramycin and mitomycin C. PMID- 6297484 TI - Induction by retinoic acid of NAD+-glycohydrolase activity of myelomonocytic cell lines HL-60, THP-1 and U-937, and fresh human acute promyelocytic leukemia cells in primary culture. PMID- 6297485 TI - A mecillinam-sensitive peptidoglycan crosslinking reaction in Escherichia coli. PMID- 6297486 TI - Elevated thymocyte norepinephrine and cyclic guanosine 3',5' monophosphate in T lymphocytes from exercised mice with coxsackievirus B3 myocarditis. PMID- 6297487 TI - Effects of amino acids and derivatives of cyclic adenosine 3', 5'-monophosphate on the production of three exoenzymes by Staphylococcus simulans biovar staphylolyticus. PMID- 6297488 TI - Expression and regulation of two steps of steroidogenesis in human Leydig cells x murine cell line clone 1D hybrids. PMID- 6297489 TI - beta-Adrenergic receptor desensitization stimulates glucose uptake in C6 rat glioma cells. PMID- 6297490 TI - Evaluation of factors responsible for the inability of insulin to antagonize lipolysis due to high concentrations of catecholamines. PMID- 6297491 TI - Complete stoichiometry of free NADPH oxidation in liver microsomes. PMID- 6297492 TI - Observation of carbon labelling in cell metabolites using proton spin echo NMR. PMID- 6297493 TI - High performance liquid chromatography of the 1,25-dihydroxyvitamin D3 receptor: its application toward resolution of receptor-monoclonal antibody complexes. PMID- 6297494 TI - 8 alpha, 9 alpha-Epoxyhexahydrocannabinol formation from delta 8 tetrahydrocannabinol by mouse liver microsomes. PMID- 6297495 TI - Effect of 6,9-deepoxy-6,9-(phenylimino)-delta 6,8-prostaglandin 1(1) (U-60,257), an inhibitor of leukotriene synthesis, on human neutrophil function. PMID- 6297496 TI - Mechanisms of oxygen activation by nitrofurantoin and relevance to its toxicity. AB - Purified ferredoxin-(cytochrome c)-NADP+ oxidoreductase and xanthine oxidase were found to catalyse the reduction of nitrofurantoin to the free radical. Under aerobic conditions, the nitrofurantoin radical underwent autoxidation to regenerate the parent compound with the concomitant production of superoxide and eventually hydrogen peroxide. The nitrofurantoin radical was also shown to react with hydrogen peroxide to generate a highly reactive species which was capable of oxidising methionine to ethylene. This active oxygen radical appeared to be identical with the crypto-OH . radical, previously proposed as being formed from the analogous reaction of the methyl viologen radical with hydrogen peroxide [R.J. Youngman and E.F. Elstner, FEBS Lett. 129, 265 (1981)]. Catalase inhibited nitrofurantoin-dependent ethylene formation in both enzyme systems, whereas superoxide dismutase was only inhibitory in the xanthine oxidase mediated reaction. Although the primary function of the respective enzyme systems is to generate the nitrofurantoin radical, the xanthine oxidase reaction is markedly more complex than that of ferredoxin-(cytochrome c)-NADP+ oxidoreductase. The differences between the two enzyme reactions appear to be due to the endogenous autoxidation of xanthine oxidase. The aerobic activation of nitrofurantoin by xanthine oxidase involved the superoxide anion as an intermediate, whereas the nitrofuran was directly reduced by ferredoxin-(cytochrome c)-NADP+ oxidoreductase without a requirement for active oxygen species. PMID- 6297497 TI - Correlation of hydrophobic character of opioid peptides with their biological activity measured in various bioassay systems. AB - Relative hydrophobicity of a series of enkephalin-like peptides has been studied by partitioning in the aqueous polymeric Ficoll-dextran biphasic system. The activities of the peptides in the guinea pig ileum, mouse vas deferens and rat brain receptor binding assay systems have been examined. The biological activities of the peptides in the two latter assays were correlated with their hydrophobic character. No correlation between the relative hydrophobicity of the compounds and their activity in the guinea pig ileum could be established. The correlations obtained appear to indicate that the drug-receptor interactions in the two bioassays occur under various conditions in regard to the local ionic composition at the membrane and/or the ionizable state of the receptor site. The activities of morphine and a number of similar drugs are described by a relationship different from that found for the peptides, which seems to support the multiple opiate receptor hypothesis. PMID- 6297498 TI - Inhibition of the cyclic AMP-adenylate cyclase system and of secretion by high concentrations of adenosine in the dog thyroid. AB - Adenosine inhibits cyclic AMP accumulation in stimulated slices and adenylate cyclase in acellular preparations of dog thyroid. The onset of this inhibition is rapid, requires relatively high adenosine concentrations (greater than or equal to 10 microM) and occurs with all activators tested (TSH, PGE1, forskolin and cholera toxin). The manganous ion, which uncouples receptor and cyclase, enhances the inhibition by adenosine. The effect of 2',5'-dideoxyadenosine, the high concentration of adenosine needed, the Mn2- effect and the lack of reversal by methylxanthines all suggest that this effect bears on the "P"-site, i.e. on the cyclase itself. Adenosine also inhibits thyroid secretion, which shows that its effect bears on the follicular cells. However the fact that cyclic AMP and DB cyclic AMP induced secretion are also reduced by adenosine suggests that adenosine also inhibits cyclic AMP action. PMID- 6297499 TI - Inhibition by nitroso-chloramphenicol of the proton translocation in mitochondria. PMID- 6297500 TI - Phosphorylation of selected substrates by semipurified cytoplasmic kinases of three P815 murine leukemias exhibiting collateral sensitivity (P815/TG), sensitivity (P815) or resistance (P815/ARA-C) to 1-beta-D arabinofuranosylcytosine. PMID- 6297501 TI - Sulfhydryl group involvement in the modulation of guanosine 3',5'-monophosphate metabolism by nitric oxide, norepinephrine, pyruvate and t-butyl hydroperoxide in minced rat lung. AB - The cyclic GMP content of rat lung mice was increased nearly 50-fold within 4 sec following exposure to nitric oxide. This rapid increase in cyclic GMP accumulation was prevented by 10 mM, but not 1 mM, dithiothreitol which itself caused a slower yet massive (100-fold) increase in the cyclic GMP content of lung mince. Tissue cyclic GMP following nitric oxide exposure declined rapidly even in the presence of the phosphodiesterase inhibitor 1-methyl-3-isobutylxanthine. The decline in cyclic GMP was accelerated by the thiol oxidant diamide (1 mM). The cyclic GMP content of lung mince was also increased by norepinephrine, pyruvate and t-butyl hydroperoxide. Diamide blocked cyclic GMP accumulation in response to these other agents as well as that caused by nitric oxide or dithiothreitol. The results suggest that sulfhydryl group modification may be a common pathway for the enhancement of cyclic GMP synthesis in tissues by a variety of stimuli. PMID- 6297502 TI - Effect of vanadium in the +5, +4 and +3 oxidation states on cardiac force of contraction, adenylate cyclase and (Na+ + K+)-ATPase activity. AB - The influence of vanadium in the nominally +5 (NH4VO3; referred to as V5+), +4 (C10H14O5V and VOSO4; V4+) and +3 oxidation states (VCl3; V3+) on cardiac force of contraction, adenylate cyclase and (Na+ + K+)-ATPase activity was investigated in order to determine which form of vanadium mediates the cardiac effects. V5+, V4+ and V3+ (300 microM each) increased the force of contraction of isolated electrically driven cat papillary muscles by about 100%. In the presence of the reducing agent ascorbic acid (5 mM) none of the three compounds led to any distinct increase in force of contraction. On the particulate adenylate cyclase preparation from feline right ventricles only V5+ stimulated the enzyme activity by about 100%, whereas V4+ and V3+ were ineffective. In the presence of 5 mM ascorbic acid all three compounds were ineffective. In contrast, in the presence of the oxidizing agent diamide (azodicarboxylic acid-bis-dimethylamide; 1 mM) all three compounds became stimulatory. On the isolated (Na+ + K+)-ATPase V5+ (500 microM) alone reduced the basal activity by about 95%. In the presence of ascorbic acid the inhibitory effect of V5+ was greatly diminished. Similar results were obtained with V4+, V3+ (100 microM) alone inhibited (Na+ + K+) ATPase activity only by about 40%. In the presence of ascorbic acid V3+ was ineffective. From the results it is concluded that positive inotropism, stimulation of adenylate cyclase and inhibition of (Na+ + K+)-ATPase by vanadium compounds likewise result from an action of vanadium in the +5 oxidation state. PMID- 6297503 TI - Multihormonal regulation of renal kallikrein. The involvement of the renin angiotensin-aldosterone system, the corticotropin-glucocorticoid system, antidiuretic hormone, catecholamines and prostaglandins. PMID- 6297504 TI - Interaction of ticlopidine with the erythrocyte membrane. AB - The membrane effects of ticlopidine on the erythrocyte membrane were explored by the spin label method at the proteic and phospholipidic levels. This spectroscopic study was completed by polyacrylamide gel electrophoresis of proteins, measurement of the protection against haemolysis and observation of the erythrocyte shape changes induced by the drug. Two types of effects have been observed. At concentrations higher than 5 x 10(-4) M, ticlopidine is a weak denaturating agent of the membrane proteins. At concentrations of pharmacological interest, the main effect of the drug is a protection against hypotonic haemolysis, and an increase in the fluidity of the membrane phospholipidic core. This last result could explain in part the interesting pharmacological effect of ticlopidine on various circulatory troubles. PMID- 6297506 TI - Selectivity of antitemplates as inhibitors of deoxyribonucleic acid polymerases. AB - DNA polymerase alpha from calf thymus was relatively insensitive to the action of partially thiolated polycytidylic acid (MPC) which had been shown previously to be a potent inhibitor of the corresponding enzyme from regenerating rat liver, competitive with the activated DNA template. In contrast, partially thiolated polyuridylic acid (MPU) strongly inhibited the calf thymus enzyme as well, but showed non-competitive kinetics with respect to the activated DNA template. The much more potent inhibitory activity of MPU compared to MPC was attributed to the less rigid conformation of the former. Methyl substitution on the 5-mercapto groups of MPU substantially decreased but did not abolish its inhibitory activity. MPU was also a potent inhibitor of the herpes virus (HSV-1) induced DNA polymerase which, too, showed little sensitivity toward MPC; in this case, the inhibition by MPU was uncompetitive with respect to the DNA template. In preliminary experiments, MPU showed significant (61%) inhibition of the replication of HSV-1, while MPC was inactive. The results demonstrate that the inhibitory activity of partially thiolated synthetic polynucleotides toward certain DNA polymerases is dependent on the base composition. PMID- 6297505 TI - Effect of N6-methyladenosine on fat-cell glucose metabolism. Evidence for two modes of action. AB - N6-Methyladenosine strongly stimulates [1-14C]glucose oxidation in rat adipocytes [J.E. Souness and V. Chagoya de Sanchez, Fedn Eur. Biochem. Soc. Lett. 125, 249 (1981)]. The effect of the adenosine analogue is largely independent of its action as an R-site agonist. Removal of endogenous adenosine was a prerequisite for the manifestation of the effect of N6-methyladenosine. Nucleoside uptake inhibitors, dipyridamole and nitrobenzylthioinosine, did not block the action of N6-methyladenosine on [1-14C]glucose oxidation. The effect of the adenosine analogue was not greatly influenced by N6-phenylisopropyladenosine, nicotinic acid or theophylline. Although N6-methyladenosine stimulated 3-O-methylglucose uptake into fat cells, it is uncertain whether this was its only effect on glucose metabolism, in view of the comparable enhancement of hexose transport elicited by N6-phenylisopropyladenosine, a much weaker stimulator of glucose oxidation. That hexose transport is not the sole site of action of N6 methyladenosine was supported by the finding that, under conditions which have been proposed to make glucose transport rate limiting, the adenosine analogue only weakly enhanced [1-14C]glucose oxidation. The conversion of glucose carbon 1 to 14CO2 was enhanced by N6-methyladenosine to a greater degree than that of carbon 6, suggesting an increase in pentose phosphate shunt activity. Mechanisms by which this could arise are discussed. Although similarities exist between the effects of insulin and N6-methyladenosine on adipocyte glucose metabolism, the mechanisms by which they stimulate glucose oxidation appear to be distinct, in view of the additivity of their actions on [1-14C]glucose conversion to 14CO2. The results indicate that N6-methyladenosine affects fat-cell glucose metabolism via two different mechanisms: (1) a weak R-site-dependent mode of action related to stimulation of glucose transport and inhibition of lipolysis, and (2) a strong R-site-independent effect of unknown mechanism. PMID- 6297507 TI - Differential release of plasminogen activator and latent collagenase from mononuclear cell-stimulated synovial cells. AB - Independent studies have previously shown that mononuclear cell supernatants stimulate the release of plasminogen activator and latent collagenase from synovial cell monolayer cultures. Simultaneous secretion of these enzymes could be an important pathway for tissue destruction under inflammatory conditions, since plasminogen activator can cause activation of latent collagenase in the presence of plasminogen. We investigated the kinetics of release of the two enzymes from synovial cells in response to the addition of mononuclear cell supernatants and retinoic acid. Synovial cells derived from osteoarthritic and rheumatoid arthritic patients responded similarly. Plasminogen activator is released within a few hours of stimulation, and secretion usually stops when the stimulus is removed. In contrast, significant amounts of collagenase are secreted only after an initial lag period of 1--2 days, and secretion is sustained long after removal of mononuclear cell supernatant. Another difference in regulation of the secretion of these two neutral proteinases is that the addition of all trans retinoic acid to the same synovial cell culture elevates plasminogen activator secretion while suppressing that of latent collagenase. Differential regulation of these enzymes under conditions of chronic inflammation may allow for continual accumulation of latent enzyme(s) which are activated during short periods of plasminogen activator release. PMID- 6297508 TI - Collagenase and collagenolytic activity in human osteoarthritic cartilage. AB - Forty-nine specimens of human cartilage were taken from 3 sites on the tibial plateau (center of osteoarthritic lesion, edge of lesion, and remote site) and graded histologically by the scale of Mankin. The tissue was homogenized and centrifuged to obtain an insoluble pellet. This was resuspended in buffer and incubated at 37 degrees C, pH 7.5. Collagen digestion was quantitated by the release of hydroxyproline-containing peptides. The highest collagenolytic activity (4.6%) was found in the center of lesions, declining in remote sites to 2.4% and in controls to 1.1%. Moderately severe disease of grade 6--9 had the highest collagenolytic activity. Approximately 55% of the metal-dependent collagenolytic activity was in a latent form, activatable by amino-phenylmercuric acetate; the remainder was self-active. A method was developed for the extraction of collagenase from cartilage; the extracted enzyme produced the typical 75:25 cleavage products of type I collagen. PMID- 6297509 TI - Rheumatoid synovial cell morphologic changes induced by a mononuclear cell factor in culture. AB - Adherent rheumatoid synovial cells in culture produce large amounts of prostaglandin E2 (PGE2) and collagenase. When exposed to a monocyte-derived factor, such cells exhibit marked increases in PGE2 and collagenase production. In addition, cellular morphology becomes more stellate. In the presence of this factor, indomethacin inhibits both PGE2 production and the stellate changes, whereas collagenase production usually continues at a high rate. Addition of PGE2 to cultures reproduces the stellate change as does the cyclic adenosine monophosphate (cAMP) analog 8-bromo-cAMP. Colchicine inhibits morphologic transformation induced by the monocyte-derived factor, whereas cytochalasin B has no effect. It appears that the stellate morphology is dependent upon PGE2-induced cAMP stimulation and is not related to collagenase production per se. PMID- 6297510 TI - Effects of gold compounds on the function of phagocytic cells. II. Inhibition of superoxide radical generation by tripeptide-activated polymorphonuclear leukocytes. AB - The effect of sodium aurothiomalate and triethylphosphine gold on the generation of superoxide radicals by chemotactic tripeptide-activated polymorphonuclear leukocytes has been investigated using a cytochrome C reduction technique. Neither gold compound inhibited the binding of the tripeptide to cells. Sodium aurothiomalate in concentrations ranging from 1 to 100 micrograms/ml produced mild, nonsignificant inhibition of the generation of superoxide radicals. In contrast, triethylphosphine gold at a concentration of 0.25 microgram/ml was associated with a mild enhancement of superoxide radical production, but the enhancement was not statistically significant. At concentrations of 1.25 micrograms/ml and above, there was a profound inhibition of superoxide radical production by these activated cells. Since the products of the respiratory burst, including superoxide, are thought to be inflammatory mediators, it is postulated that the inhibition of superoxide radical generation by gold compounds may be another mechanism by which the compounds modulate their antiinflammatory effects in patients with rheumatoid disease. PMID- 6297511 TI - Effects of dietary enrichment with eicosapentaenoic acid upon autoimmune nephritis in female NZB X NZW/F1 mice. AB - A menhaden oil diet, rich in eicosapentaenoic acid, protected female NZB X NZW/F1 mice from autoimmune nephritis. Only 15% of mice treated with the diet from weaning had died with severe renal disease at 19 months, versus 98% of controls on a beef tallow diet. The menhaden oil also protected these mice from renal disease when instituted at 4 and 5 months of age and, under these conditions, levels of anti-native DNA antibodies were similar in both dietary groups. Our data suggest that the menhaden oil diet may act primarily to reduce inflammation via the ability of eicosapentaenoic acid to alter the production of prostaglandins and leukotrienes. PMID- 6297512 TI - Cervical radiculopathy: a rare symptom of giant cell arteritis. PMID- 6297513 TI - [Enzymatic detoxication of organophosphorus insecticides and nerve gases in primates]. AB - The detoxication of organophosphorus compounds by phosphorylphosphatases was studied in primates. Taking into account the distribution of paraoxonase (EC 3.1.1.2) and DFPase (EC 3.8.2.1) in different tissues of the monkey (Macaca mulatta), the total detoxicating capacity for diethyl-p-nitrophenylphosphate (paraoxon, E 600) and diisopropylphosphorofluoridate (DFP) was determined. Acetylcholinesterase (AChE) (EC 3.1.1.7) of human brain was inhibited in vitro by paraoxon and DFP. Using the rate constants of AChE-inhibition and of AChE synthesis those concentrations of organophosphorus inhibitors were calculated, which in vivo would reduce the steady-state AChE-activity to 20% of normal. This acute ineffective concentration is 7.6 X 10(-8) g/kg for DFP and 2.3 X 10(-8) g/kg for paraoxon. From substrate kinetics of the phosphorylphosphatases the time course of paraoxon and DFP detoxication in primates could be calculated. The time needed by phosphorylphosphatases to reduce a certain dose of an organophosphorus compound to the acute ineffective concentration is referred to as "effective detoxication time". The effective detoxication time (teff) was determined for different concentrations of paraoxon and DFP and was compared with the time needed by these organophosphate concentrations to inhibit AChE-activity to 12.5% of normal (t1/8). The significance of in vitro data for the evaluation of dose limits of organophosphate toxicity in vivo is discussed. PMID- 6297514 TI - Effects of 4-chloro-2-(2-imidazolin-2-ylamino)-isoindoline Hydrochloride (BE 6143) at pre- and postsynaptic alpha-adrenoceptors in rabbit aorta and pulmonary artery. AB - Strips of rabbit aorta and pulmonary artery were used to study the effects of 4 chloro-2-(2-imidazolin-2-ylamino)-isoindoline hydrochloride (BE 6143 or BDF 6143) on pre- and postsynaptic alpha-adrenoceptors. BE 6143 caused contraction of both preparations with a potency slightly higher than and an intrinsic activity 0.7 0.8 that of phenylephrine. The effect was competitively antagonized by prazosin and rauwolscine. Conversely, BE 6143 antagonized contractile responses to phenylephrine, alpha-methylnoradrenaline and noradrenaline (norepinephrine) with pA2 values of 7.1-7.4. In strips preincubated with 3H-noradrenaline and then superfused, BE 6143 increased the electrically evoked overflow of tritium. Increases by 30% were produced by concentrations of about 1.5 nmol/l; high concentrations failed to change or even reduced the evoked overflow. BE 614 0.01 mumol/l also increased electrically evoked contractions whereas high concentrations were inhibitory. The drug shifted the log concentration-response curve of clonidine for inhibition of evoked tritium overflow to the right in parallel manner; from the shift, a pA2 value of 8.3 was calculated. When presynaptic alpha-adrenoceptors had been blocked by yohimbine or rauwolscine, BE 6143 0.01-10 mumol/l caused a decrease in evoked tritium overflow which was more pronounced, the higher the concentration of yohimbine or rauwolscine. It is concluded that BE 6143 acts as a partial agonist at the postsynaptic alpha 1 adrenoceptors and as an antagonist at the presynaptic alpha 2-receptors of rabbit aorta and pulmonary artery. It has higher affinity pre- than postsynaptically, and its affinity for the presynaptic receptors exceeds that of yohimbine and rauwolscine. In addition, however, BE 6143 inhibits the release of noradrenaline by a non-alpha-adrenergic mechanism. PMID- 6297515 TI - Effect of a modified guar gum preparation on glucose and lipid levels in diabetics and healthy volunteers. AB - Six healthy volunteers and 17 diabetics (6 insulin-dependent and 11 diet- and tablet-treated) were treated with a special processed, palatable guar gum (10 g b.i.d. immediately before meals) for periods of one or three weeks or, in some cases, up to 13 weeks. A standardized test meal was given to study the effect of the fiber on postprandial glucose levels. Ten g guar was stirred in water and taken immediately before the test meal. The postprandial blood glucose levels were similar in the healthy volunteers but significantly lower in the diabetics following treatment with guar for one and three weeks, respectively. Furthermore, the fasting blood glucose levels were significantly lower in the diabetics after three, but not one, weeks of treatment. The lower postprandial glucose levels were coupled with attenuated and delayed insulin levels in accordance with an effect of guar gum on the rate of carbohydrate absorption. The cholesterol levels were on average reduced with 14% in the diabetics following three weeks' treatment with guar. The higher the initial cholesterol level, the greater the reduction in cholesterol; 26% reduction was achieved in four patients with initial levels above 7 mM. The alpha-lipoprotein cholesterol levels were not significantly changed, thus an increase in the alpha-lipoprotein cholesterol/total serum cholesterol ratio was obtained. Neither plasma triglycerides nor body weights altered during treatment. The reported side effects were as expected and were usually mild and transient (e.g. increased flatulence). The data show that guar gum also reduces postprandial glucose levels on a long-term basis and may improve the diabetic control. Additionally, treatment with this fiber leads to a concentration-dependent decrease in cholesterol levels. PMID- 6297516 TI - Long-term treatment of hypercholesterolaemia with a new palatable formulation of guar gum. AB - Nineteen patients with primary hypercholesterolaemia previously stabilized on diet alone were treated with a new formulation of guar gum (6g t.d.s. with meals) in a placebo-controlled, single-blind study. Seventeen patients completed 3 months treatment without serious side effects, while 2 patients withdrew immediately because of severe diarrhoea. Thirteen patients have completed 12 months treatment with guar gum. There have been no significant changes in safety parameters. Plasma cholesterol was reduced by a significant 15% during the first 3 months of treatment (7.9 +/- 0.8 vs 6.7 +/- 1.0 mmol/l, P less than 0.001) and this effect has been sustained for 12 months. The fall in plasma cholesterol was associated with a significant 20% fall in LDL cholesterol, but with no change in HDL cholesterol. Plasma triglycerides did not change significantly. Percentage cholesterol absorption was reduced by guar gum in 4/5 normal subjects examined. PMID- 6297518 TI - Saponins and fibre. Lack of interactive effects on serum and liver cholesterol in rats and hamsters. PMID- 6297517 TI - Comparative beneficial effects on platelet functions and atherosclerosis of dietary linoleic and gamma-linolenic acids in the rabbit. AB - Male rabbits (10 weeks old) were fed, for 20 weeks, purified diets rich in fat (45% of calories) containing saturated fats (butter), polyunsaturated fats (evening primrose oil + butter or sunflower oil + butter) for 20 weeks. Linoleic acid represented respectively 3.6, 33 and 34% of the dietary fatty acids, while gamma-linolenic acid was present (4.4%) solely in the second group. A significant increase in di-homo-gamma-linolenic acid in plasma, platelets and aorta was noted only in the animals fed evening primrose oil. Despite this, the results of the platelet aggregation to thrombin and ADP, the recalcification plasma-clotting time (platelet-rich plasma) and the severity of atherosclerosis were not significantly different from those observed in the group fed sunflower oil. In contrast, in comparison to the butter-fed animals, the two groups fed the polyunsaturated fats showed remarkable improvements in the clotting time (P less than 0.01) and in the severity of atherosclerotic lesions (evening primrose oil P less than 0.001; sunflower oil P less than 0.05). However, the response to thrombin-induced aggregation was significantly decreased (P less than 0.05) only in the evening primrose oil-fed animals. In these long-term studies in young rabbits, dietary gamma-linolenic acid did not seem to have marked beneficial effects, additional to those induced by linoleic acid, on platelet functions or on atherosclerosis. PMID- 6297519 TI - [Spironolactone bodies. Light and electron microscope study apropos of 4 cases of adrenal adenomas with primary hyperaldosteronism]. AB - So-called Spironolactone bodies are reported in four adrenal cortical adenomas that were removed from patients with primary hyperaldosteronism, treated preoperatively with spironolactone. Their morphology is described optically : these round acidophilic cytoplasmic bodies have a laminated appearance that ranges in size from 2 to 20 mu in diameter. Electron microscopy of the zona glomerulosa non adenomatous cells and of adenomatous cells of glomerular type reveals numerous membranes arranged concentrically around a central core. Connections between the endoplasmic reticulum and mitochondria in the cytoplasm of the cell and lining layers of the body are described and analysed. The authors attempt to explain the formation and the increase of the bodies. Correlations appear between the number of spironolactone bodies and duration of administration of spironolactone and total drug dosage. The results are discussed with those of the literature. PMID- 6297520 TI - [Deposits of unusual structure in lupus glomerulonephritis]. AB - The authors report a case of Lupus nephritis shown by ultrastructural analysis of subendothelial, mesangial and especially subepithelial deposits formed by aggregates of electron-dense straight tubular structures 20 to 30 nm in diameter. These unusual structural deposits, always identical, are observed in 4 biopsies performed over 8 years during the management of Systemic Lupus Erythematosus in a young woman. They are compared to the usual "Fingerprint" deposits of Systemic Lupus Erythematosus and to the geometric type deposits of cryoglobulinemic glomerulonephritis. PMID- 6297521 TI - Substantia nigra as a station that not only transmits, but also transforms, incoming signals for its behavioural expression: striatal dopamine and GABA mediated responses of pars reticulata neurons. PMID- 6297522 TI - [Ca++ dependence of the contraction of coronary artery segments in response to norepinephrine after beta-adrenergic blockade]. AB - The effect of external calcium concentration on the NE-induced contraction after beta-adrenergic blocking was studied in vitro. It resulted that the effect of NE was enhanced by increase, or reduced by decrease of calcium concentration. NE induced contraction was not abolished when the bathing fluid was Ca++-free. The disappearance of the NE effect was only obtained in preparations treated with EDTA and perfused with Ca++-free Ringer-Locke solution. It is concluded that NE induced contraction after beta-adrenergic blocking is Ca++-dependent and on the tissue bound Ca++. PMID- 6297523 TI - [Use of diafiltration in the study of interactions between technetium (95mTcO4-) and albumin]. AB - Reversible binding of pertechnetate (95mTcO-4) by human serum albumin (HSA) has been investigated by "wash-in" experiments, in the presence of a large excess of HSA. Equilibrium constants have been determined in the range of temperature between 5 and 37 C. Thermodynamic parameters of the process have been calculated. The marked influence of temperature on the association reaction should be taken into consideration when carrying out investigations on the metabolism of technetium in poikilotherms. PMID- 6297524 TI - [Regulation of the functions of human leukocytes by oxametacine]. AB - Non steroidal anti-inflammatory drugs, such as oxametacine, are generally used in treatment of rheumatoid disease. In an 'in vitro' experimental model, the drug efficacy was tested on leukocyte functions. Locomotion, both random and directional, phagocytic activity and superoxide production of normal and rheumatoid PMNL were tested in the presence of varying concentrations of oxometacine. Locomotion was evaluated by using modified Boyden chambers; phagocytosis was tested by number of yeast particles injested and by NBT reduction; superoxide production was assayed by reduction of ferricytochrome C. In our conditions the drug exhibited a strong anti-inflammatory effect. In fact, chemotaxis and anion production were specifically depressed in a dose-dependent way. PMID- 6297525 TI - [Radioisotope study of left ventricular aneurysms]. PMID- 6297526 TI - [Biochemical basis of the electrophysiological mechanisms in arrhythmias in prior subjection to therapy]. PMID- 6297527 TI - Reorientation of prostaglandin endoperoxide metabolism by a thromboxane synthetase inhibitor: in vitro and clinical observations. AB - 1 Work with dazoxiben in vitro and in vivo suggests that a diminished capacity of platelets to synthesize thromboxane A2 results in a reorientation of the metabolism of cyclic endoperoxides. 2 Platelets of patients with congenital thromboxane synthetase deficiency show the same phenomenon. 3 In the presence of endothelium or leukocytes which have prostacyclin synthetase capacity, significant amounts of prostacyclin can be generated if thromboxane synthesis is blocked. 4 The local generation of aggregation inhibiting prostaglandins in areas of vascular damage may be an interesting therapeutic concept. 5 Pilot clinical studies of thromboxane synthetase inhibitor dazoxiben (UK 37248) in a small number of patients with peripheral arterial disease did not reveal major or consistent haemodynamic changes. 6 Attention is directed towards optimizing pharmacologically the generation and efficacy of the inhibitory prostaglandins produced when thromboxane synthetase is inhibited. PMID- 6297528 TI - Assay of anticancer drugs in tissue culture: cell cultures of biopsies from human astrocytoma. AB - A method has been developed for measuring the drug sensitivity of human gliomas in short-term culture, using scintillation counting or autofluorography. Cell cultures prepared from malignant astrocytomas were treated with anticancer drugs whilst in exponential growth in microtitration plates. After drug treatment and a recovery period, residual viability was measured by [3H] leucine incorporation followed by scintillation counting or by [35S] methionine incorporation and autofluorography in situ. In 5 glioma cell lines tested against 6 drugs, the microtitration method correlated well with monolayer cloning. Although replicate samples of the same tumour showed little variation in chemosensitivity, there was marked variation between the chemosensitivities of cultures derived from the tumours of different patients. However, as variability between replicates was apparent during drug exposure or shortly after, it is important to allow the assay to run as long as possible after drug removal. It is hoped that this assay may provide the basis of a method for the prediction of in vivo chemosensitivity or the screening of potential chemotherapeutic drugs. PMID- 6297529 TI - High dose cyclophosphamide treatment of human oat cell xenografts in immune deprived mice. AB - Immunodeprived mice survived a high, otherwise lethal dose of cyclophosphamide (Cy) provided they had been "primed" with a low dose (50 mg kg-1) of the drug 4 days earlier. These combinations were then tested on 2 oat cell xenograft lines (which are known to reproduce the chemotherapeutic responses of the parent tumours) grown in immunodeprived mice. In the treatment of the first oat cell xenograft, 200 mg kg-1 Cy produced a growth delay of 34 days in the unprimed group and 45 days in the primed group. At a dose of 300 mg kg-1 a growth delay could not be assessed in the control group as 16/17 of these unprimed mice bearing this xenograft died. However, 14/22 tumours went into complete remission in this group before death occurred. In contrast only 3/16 deaths occurred in the group of mice that were primed before receiving the same challenge dose. In these animals 19/26 tumours went into complete remission and were still completely absent when the experiment was terminated at 60 days. Using the second oat cell xenograft, 300 mg kg-1 Cy produced a growth delay of 27 days. However, at this dose level all the animals were dead by day 46. In mice which had been primed with 50 mg kg-1 Cy 4 days before the administration of 300 mg kg-1 a growth delay of 32 days was achieved and 2/9 animals were alive at day 60. This study shows that priming allows larger doses of Cy to be given to immunodeprived mice bearing human tumour xenografts than would normally be tolerated and that the priming does not alter the anti-tumour efficacy of the large challenge dose as measured by tumour growth delay or complete remission rate. As the tumours were human in origin it raises the question whether high dose cyclophosphamide therapy and priming have a role to play in the treatment of patients with oat cell carcinoma. PMID- 6297530 TI - Thalassaemia intermedia in Cyprus: the interaction of alpha and beta thalassaemia. AB - Restriction endonuclease analysis has been performed on the alpha and beta globin gene clusters of 57 Cypriots homozygous for beta thalassaemia, 30 with the transfusion dependent form of the condition (thalassaemia major) and 27 who are less severely affected (thalassaemia intermedia). There was a significant difference in the incidence of alpha thalassaemia between the two groups: 14/27 of the patients with thalassaemia intermedia also had deletion forms of alpha thalassaemia, while only 4/30 of the patients with thalassaemia major were similarly affected. Thus in Cypriot patients who are homozygous for beta thalassaemia the co-inheritance of alpha thalassaemia is an important factor in determining the clinical course. PMID- 6297531 TI - 5-nucleotidase activity of megakaryoblasts in a case of acute megakaryoblastic leukaemia. PMID- 6297532 TI - Mortality of workers manufacturing friction materials using asbestos. AB - A mortality (1942-80) study was carried out on 13460 workers of a factory producing friction materials. The only type of asbestos used was chrysotile, except during two well-defined periods before 1945 when crocidolite was used, and over 99% of the population was traced. Compared with national death rates there were no detectable excesses of deaths due to lung cancer, gastrointestinal cancer, or other cancers; 11 deaths were due to pleural mesothelioma. A case control study was carried out on deaths due to mesothelioma; this showed that eight workers had been exposed to crocidolite and another was possibly exposed intermittently to crocidolite. The other two had been employed for most of their working lives outside the factory, and their mesotheliomas could not be definitely attributed to exposure to chrysotile. Limiting the study to cases and controls who had exposure to 5 fibres/ml of chrysotile asbestos it was found that five of the six cases compared with two of the 10 controls had also been exposed to crocidolite. The probability (1:36) of this occurring were there no association with crocidolite is most unlikely. A case-control study was also carried out on deaths due to lung cancer and gastrointestinal cancer to investigate the dose-response relationships between these tumours and exposure to chrysotile. Measured and estimated fibre concentrations were available for the different jobs over the period of the study. No dose-response relationships were observed, but the exposures were low with only 5% of men accumulating 100 fibre years/ml. The experience at this factory over a 40-year period showed that chrysotile asbestos was processed with no detectable excess mortality. PMID- 6297533 TI - Complement factors in workers exposed to silicosis. PMID- 6297534 TI - Environmental history of a factory producing friction material. AB - The fibre concentrations generated during the production of friction materials, incorporating asbestos, over the past 60 years have been studied to provide cumulative dust exposure data for a mortality study of the work-force. Chrysotile has been used almost exclusively during this period. The measurements made routinely by the factory staff provided information from 1950 onwards; concentrations for earlier years were derived from simulation studies using original materials, machines, and methods. These have shown that while high concentrations prevailed in the earliest years dust suppression measures initiated in 1931, and other factors, reduced the time-weighted average concentrations to moderate levels. There were two four-year periods when the production of brake blocks incorporating crocidolite occupied a well-defined area of the factory. The simulation of this production was not permissible, but the employees concerned could be identified. PMID- 6297535 TI - Cervical and serum IgA and serum IgG antibodies to Chlamydia trachomatis and herpes simplex virus in threatened abortion: a prospective study. AB - The aetiological role of Chlamydia trachomatis (CT) and herpes simplex virus (HSV) was investigated in 189 patients with threatened abortion. Assessment of infection was based on isolation, and on determination of serum immunoglobulin (Ig)G and IgA antibodies as well as cervical IgA antibody levels with new sensitive radioimmunoassay (RIA) techniques. One third of the women were delivered of a healthy infant and two thirds aborted, but the two groups were otherwise clinically similar. By isolation, only 2.7% of the patients were CT positive, but increased cervical IgA antibody level to CT was detected in 41.3%. The mean level of these local antibodies was similar in both study groups, but the mean levels of serum IgA and IgG antibodies were somewhat higher in the patients who aborted although the difference was not significant. None of the cervical specimens was positive for HSV by isolation but the cervical IgA antibody level to HSV was raised in 47.1% of the patients. Both cervical and serum IgA antibody levels to HSV were significantly raised among the patients who aborted, but there were no differences between the patients with spontaneous abortion and those with a blighted ovum. There was no clear association between CT and abortion, but an association between HSV and abortion is possible. The incidence of raised levels of both CT and HSV IgA antibodies in the cervix was surprisingly high in both groups and the significance of this finding remains to be investigated. PMID- 6297536 TI - Lipids and lipoproteins in women after oophorectomy and the response to oestrogen therapy. AB - The short-term effects of different types and doses of oestrogen on serum lipids and lipoproteins were studied in 35 oophorectomized women. After 3 months treatment, serum cholesterol levels were unaffected by 1 and 2 mg of micronized 17 beta-oestradiol or 0.625 and 1.25 mg of conjugated equine oestrogens. Triglyceride levels were significantly elevated after treatment with 1.25 mg of conjugated oestrogens. A trend towards a higher relative proportion of high density lipoproteins and a lower relative proportion of low-density lipoproteins was observed for all four oestrogen regimens, however, statistical significance was not achieved. The proportion of very-low-density lipoprotein was unaffected by oestrogen treatment. The age of the oophorectomized women was found to have no effect on either the direction or magnitude of the lipid or lipoprotein responses to oestrogen. Using FSH depression as an index, 1.25 mg of conjugated oestrogens was found to be the most potent of the four oestrogen regimens tested. Therefore, with respect to lipid balance, little additional clinical benefit is achieved by using a more potent regimen and the risk of adverse side effects may be increased. PMID- 6297537 TI - Serum and tear immunoglobulin levels in acute adenovirus conjunctivitis. AB - Tear and serum IgG, IgA, and IgM levels were measured in 35 healthy subjects and 31 patients with acute conjunctivitis due to adenovirus 2, 7, and 8 infections. Serum IgG and IgM levels were found to be significantly low and there was no change in the serum IgA level. There was a significant rise in the tear IgG level. A progressive decline in the serum immunoglobulin level with the severity of the disease was observed. The changes in the immunoglobulin levels in serum and tears showed a tendency to reach normal levels with clinical improvement. PMID- 6297538 TI - Human serum thymidine triphosphate nucleotidohydrolase: purification and properties of a new enzyme. AB - Thymidine triphosphate monophosphohydrolase (dTTPase), an enzyme which catalyzes the hydrolysis of dTTP to the corresponding diphosphate (dTDP), has been purified to homogeneity from human serum. The enzyme sediments with 3.8 S in sucrose density gradients. A Stokes radius of 31 A is estimated by gel filtration. Accordingly, its molecular weight is 48 500. Since only one single band of Mr 24 000 is detected after sodium dodecyl sulfate/polyacrylamide gel electrophoresis, the native enzyme seems to be composed of two identical subunits. The enzyme exhibits optimal activity over a pH range from 7 to 9, and the activation energy is estimated to be 7.1 kcal/mol (29.7 kJ/mol) at pH 7.8. While the enzyme is active in the absence of added divalent cations, the activity can be inhibited by ethylenediaminetetracetic acid (EDTA) but not by phenanthroline. The inhibition caused by EDTA is reversed by Mn2+. Zn2+ causes a complete inhibition of enzyme activity. No requirement exists for a sulfhydryl compound. The enzyme has an Rf value of 0.45, an isoelectric point of 5.2, and an apparent Km value of 40 microM for dTTP. dUTP and UTP are degraded by about 50 and 20% of the rate of dTTP hydrolysis, respectively. Other deoxyribonucleosides or ribonucleoside triphosphates do not serve as substrates for the dTTPase. The existence of this enzyme is significant since it could play a role in the regulation of the cellular dTTP levels. PMID- 6297539 TI - Enzyme digestion of intermediates of excision repair in human cells irradiated with ultraviolet light. PMID- 6297540 TI - Evidence for modulation of the heme absorptions of cytochrome c oxidase by metal metal interactions. PMID- 6297541 TI - Membrane-associated purine metabolizing enzyme activities of human peripheral blood cells. AB - Sealed and unsealed plasma membrane vesicles were prepared from human erythrocytes and lymphocytes. Phosphoribosylpyrophosphate synthetase (PRibPP synthetase), hypoxanthine phosphoribosyltransferase (HPRTase), and adenine phosphoribosyltransferase (APRTase) activities are detectable on both inside-out and right-side-out sealed vesicles. Ghost preparations were about 0.2%, 1%, and 1.2% of the total erythrocyte and 0.5%, 5.3%, and 9.7% of the lymphocyte APRTase, HPRTase, and PRibPP synthetase activities. The rapid decrease in these enzyme activities, upon further purification of the membranes, seemed to suggest that they might be loosely bound extrinsic proteins. Evidence confirming the localization of these enzymes on the cell surface was obtained by measuring production of [14C]AMP by intact cells in medium containing [14C]adenine, ribose 5-phosphate, and Mg2+ATP. The formation of AMP was linear with time and number of cells present. Magnesium and phosphate exerted different effects on the production of extracellular AMP than on intracellular, which involves transport as well as phosphoribosylation. Cytosoluble and membrane-bound APRTase and PRibPP synthetase exhibited different catalytic properties and sensitivities to effectors. Membranes of erythrocytes of HPRTase-deficient patients contain little or no HPRTase activity when assayed in the absence of Triton. Reisolation of these membranes from admixture with normal hemolysates did not result in any bound activity; thus, the membrane-bound activity is not an artifact of the isolation procedure. Lysis with Triton released activity equal to about half that of control membranes. This is further evidence that the enzyme is firmly bound to the membrane. PMID- 6297542 TI - Phosphorus-31 nuclear magnetic resonance studies of the conformation of an adenosine 5'-triphosphate analogue at the active site of (Na+ + K+)-ATPase from kidney medulla. AB - It has previously been shown that there are two sites for divalent metals at the active site of kidney (Na+ + K+)-ATPase, one bound directly to the enzyme and one coordinated to the ATP substrate [Grisham, C. (1981) J. Inorg. Biochem. 14, 45; O'Connor, S., & Grisham, C. (1980) FEBS Lett. 118, 303]. The conformation of the metal-nucleotide complex has been studied by using beta, gamma-bidentate Co (NH3)4ATP, a substitution-inert analogue of MgATP. Kinetic studies show that Co(NH3)4ATP is a competitive inhibitor with respect to MnATP for the (Na+ + K+) ATPase. The Ki values under both high- and low-affinity conditions (Ki = 10 microM and Ki = 1.6 mM, respectively) are similar to the Km values for MnATP under the same conditions (2.88 microM and 0.902 mM). From the paramagnetic effect of Mn2+ bound to the ATPase on the longitudinal relaxation rates of the phosphorus nuclei of Co(NH3)4ATP at the substrate site (at 40.5 and 145.75 MHz), Mn-P distances to all three phosphates are determined. The distances are consistent with the formation of a second sphere coordination complex on the enzyme between Mn2+ and the phosphates of Co(NH3)4ATP. In this respect, kidney (Na+ + K+)-ATPase appears to be similar to pyruvate kinase [Sloan, D., & Mildvan, A. (1976) J. Biol. Chem. 251, 2412] and phosphoribosylpyrophosphate synthetase [Granot, J., Gibson, K., Switzer, R., & Mildvan, A. (1980) J. Biol. Chem. 255, 10931]. Roles for both of the active site divalent cations are discussed. PMID- 6297543 TI - 23-Keto-25-hydroxyvitamin D3: a vitamin D3 metabolite with high affinity for the 1,25-dihydroxyvitamin D specific cytosol receptor. AB - A new metabolite of 23,25-dihydroxyvitamin D3 has been generated with kidney homogenates prepared from vitamin D treated chicks. The metabolite was purified with three high-performance liquid chromatographic steps and was identified as 23 keto-25-hydroxyvitamin D3 by ultraviolet absorption spectroscopy, mass spectrometry, and chemical reactivity. The R stereoisomer of 23,25 dihydroxyvitamin D3 was 10-fold more effective as an in vitro precursor to 23 keto-25-hydroxyvitamin D3 than was the naturally occurring S stereoisomer. Approximately 500 ng of 23-keto-25-hydroxyvitamin D3 was necessary to produce the same degree of intestinal-calcium transport as 25 ng of vitamin D3--a difference of about 20-fold. 23-Keto-25-hydroxyvitamin D3 was not active at stimulating bone calcium resorption at the doses and times tested. This new vitamin D3 metabolite, however, had greater affinity than 25-hydroxyvitamin D3 to both the rat plasma vitamin D binding protein and the 1,25-dihydroxyvitamin D specific cytosol receptor. Heretofore, only 1 alpha-hydroxylated metabolites of 25-hydroxyvitamin D3 or analogues possessing a pseudo 1 alpha-hydroxy group were known to bind to the 1,25-dihydroxyvitamin D receptor with higher affinity than 25-hydroxyvitamin D3. Ketone formation at the 23 position, therefore, is the first side-chain modification of 25-hydroxyvitamin D3 that results in enhanced binding to the 1,25 dihydroxyvitamin D receptor binding protein. PMID- 6297544 TI - Characterization of deoxyribonucleic acid sequences at the 5' and 3' borders of the 100 kilobase pair ovalbumin gene domain. PMID- 6297545 TI - Interaction of cytochrome c with reaction centers of Rhodopseudomonas sphaeroides R-26: localization of the binding site by chemical cross-linking and immunochemical studies. AB - The location of the cytochrome binding site on the reaction center of Rhodopseudomonas sphaeroides was studied by two different approaches. In one, cross-linking agents, principally dithiobis(propionimidate) and dimethyl suberimidate, were used to link cytochrome c and cytochrome c2 to reaction centers; in the other, the inhibition of electron transfer by antibodies against the subunits was investigated. Cytochrome c (horse) cross-linked to the L and M subunits, whereas cytochrome c2 (R. sphaeroides) cross-linked only to the L subunit. The cross-linked reaction center-cytochrome complexes were isolated by affinity chromatography. The rate of electron transfer in the cross-linked cytochrome c2 complex was the same as that in the un-cross-linked complex. However, when cytochrome c was used, the rate in the cross-linked complex was about 15 times slower than that in the un-cross-linked complex. Fab fragments of antibodies specific against the L and M subunits blocked electron transfer from both cytochrome c (horse) and cytochrome c2 (R. sphaeroides). Antibodies specific for the H subunit did not block either reaction. We conclude that the cytochrome binding site on the reaction center is close (approximately 10 A) to both the L and M subunits, possibly in a cleft between them. PMID- 6297546 TI - Binding subsites in human thrombins. PMID- 6297547 TI - Characterization of complexes between Escherichia coli sulfite reductase hemoprotein subunit and its substrates sulfite and nitrite. PMID- 6297548 TI - A differential polarized phase fluorometric study of the effects of high hydrostatic pressure upon the fluidity of cellular membranes. AB - The effects of high hydrostatic pressure (up to 2 kbar) upon the fluidity and order of the synaptic and myelin membrane fractions of goldfish brain have been studied by using steady-state and differential polarized phase fluorometry. Probe motion provided a measure of membrane order (r infinity) and probe rotational rate (R). Membrane order became progressively greater as pressure was increased up to approximately 2 kbar. This effect was similar over the temperature range 5.6-34.3 degrees C. An increase in pressure of 1 kbar had an effect on membrane order that was equivalent to a 13-19 degrees C reduction in temperature. Membrane order was essentially identical during pressurization and depressurization. At 5.6 degrees C, pressurization caused a large increase in R, and similar, though less dramatic, anomalies occurred at higher temperatures. It is suggested that this is due to the segregation of probe molecules in highly ordered membranes, which leads either to excitation transfer between 1,6-diphenyl-1,3,5-hexatriene (DPH) molecules or to changes in the rotational motion of DPH from "sticking" to "slipping". PMID- 6297550 TI - Origin of the cytochrome a absorption red shift: a pH-dependent interaction between its heme a formyl and protein in cytochrome oxidase. PMID- 6297549 TI - Covalent labeling of the acetylcholine receptor from Torpedo electric tissue with the channel blocker [3H]triphenylmethylphosphonium by ultraviolet irradiation. AB - The lipophilic cation [3H]triphenylmethylphosphonium, frequently used as a voltage sensor in membrane systems, binds reversibly to a site different from the acetylcholine binding site. This is concluded from the different pH dependences of the binding of these two ligands. Furthermore [3H]triphenylmethylphosphonium, previously identified as a channel blocker, can be covalently incorporated into acetylcholine receptor-rich membranes from Torpedo electric tissue by UV irradiation of the receptor-ligand complex. In the absence of effector, predominantly the alpha-polypeptide chains (Mr 40000) of the receptor protein are labeled by the radioactive ligand. The agonist carbamoylcholine strongly stimulates the labeling, but it directs the label predominantly to the delta- and beta-polypeptide chains. The antagonist D-tubocurarine and the virtually irreversible competitive antagonist alpha-bungarotoxin have qualitatively the same effect as the agonist carbamoylcholine. Significant differences were obtained with receptor-rich membranes prepared from Torpedo marmorata and Torpedo californica: No agonist- or antagonist-stimulated reaction was observed with the latter. The results are interpreted as an indication of a rearrangement of the receptor's quaternary structure caused by cholinergic effector binding preceding discrimination between agonists and antagonists. PMID- 6297551 TI - Photolabeling approach to the study of the topography of the atractyloside binding site in mitochondrial adenosine 5'-diphosphate/adenosine 5'-triphosphate carrier protein. PMID- 6297553 TI - New insights, ideas and unanswered questions concerning iron-sulfur clusters in mitochondria. PMID- 6297552 TI - Electron paramagnetic resonance and optical evidence for interaction between siroheme and Fe4S4 prosthetic groups in complexes of Escherichia coli sulfite reductase hemoprotein with added ligands. AB - Janick & Siegel [Janick, P. A., & Siegel, L. M. (1982) Biochemistry 21, 3538 3547] showed that the EPR spectrum of the reduced Fe4S4 center (S = 1/2) in fully reduced native ("unligated") Escherichia coli NADPH-sulfite reductase hemoprotein subunit (SiR-HP) is perturbed by interaction with paramagnetic ferrous siroheme (S = 1 or 2) to yield several novel sets of EPR signals: one set with all g values between 2.0 and 2.8, termed "S = 1/2" type, and two sets with the lowest field g value between 4.7 and 5.4, termed "S = 3/2" type. The present study has shown that EPR spectra of fully reduced SiR-HP are nearly quantitatively converted to the classical "g = 1.94" type typical of S = 1/2 Fe4S4 clusters when the heme has been ligated by strong field ligands such as CO, CN-, S2-, and AsO2 , converting the ferroheme to S = 0. However, the exact line shapes and g values of the g = 1.94 differ markedly when different ligands are bound to the heme. Also, optical difference spectra taken between enzyme species in which the heme is kept in the same (Fe2+) oxidation state while the Fe4S4 center is reduced or oxidized show that the optical spectrum of the ligated siroheme is sensitive to the oxidation state of the Fe4S4 cluster. These results indicate that the heme Fe4S4 interaction of native SiR-HP persists even when the heme Fe is bound to exogenous ligands. We have also found that the g values of the exchange-coupled S = 1/2 and S V 3/2 type signals of native reduced SiR-HP can be significantly shifted by addition of potential weak field heme ligands--halides and formate--or low concentrations of certain chaotropic agents--guanidinium salts and dimethyl sulfoxide--to the fully reduced enzyme. Such agents can also promote interconversion of the S = 1/2 and S = 3/2 type signals. These effects are reversed on removal of the agent. Treatment of reduced SiR-HP with relatively large concentrations of chaotropes, e.g., 60% dimethyl sulfoxide or 2 or 3 M urea, leads to abolition of the S = 1/2 and S = 3/2 EPR signals and their replacement by signals of the g = 1.94 type. PMID- 6297554 TI - Fractionation of protein components of plasma membranes from the electric organ of Torpedo marmorata. AB - A procedure has been developed for the separation of intrinsic proteins of plasma membranes from the electric organ of Torpedo marmorata. (Na+ + K+)-ATPase, nicotinic acetylcholine receptor and acetylcholinesterase remained active after solubilization with the nonionic detergent dodecyl octaethylene glycol monoether (C12E8). These components could be separated by ion exchange chromatography on DEAE-Sephadex A-25. Fractions enriched in ouabain-sensitive K+-phosphatase or (Na+ + K+)-ATPase activity showed two bands in sodium dodecyl sulphate polyacrylamide gel electrophoresis corresponding to the alpha- and beta-subunits. The (Na+ + K+)-ATPase was shown to have immunological determinants in common with a 93 kDa polypeptide which copurified with the nicotinic acetylcholine receptor, also after solubilization in Triton X-100 and chromatography on Naja naja siamensis alpha-toxin-Sepharose columns. The data suggest that the alpha-subunit of (Na+ + K+)-ATPase associates with the acetylcholine receptor in the membranes of the electric organ. PMID- 6297555 TI - Cholesterol as activator of ADP-ATP exchange in reconstituted liposomes and in mitochondria. AB - The influence of cholesterol on ADP-ATP exchange activity was measured in the reconstituted system, submitochondrial (sonic) particles and mitoplasts (isolated inner mitochondrial membranes). In the reconstituted system, cholesterol markedly enhanced the nucleotide-uptake rate, when added to membranes of various compositions i.e., pure phosphatidylcholine, phosphatidylcholine/phosphatidylethanolamine mixtures and crude egg yolk phospholipids. The stimulation was linearly dependent on the amount of incorporated cholesterol up to 7-13% added sterol, depending on the type of phospholipids. Cholesterol influenced neither the amount of actively reconstituted carrier proteins nor the affinity of the carrier towards nucleotides nor the breakpoint of temperature dependence in the Arrhenius plot. The stimulation could be correlated with an increase in the molecular activity of the carrier protein. The influence of cholesterol was also measured in the natural environment of the carrier protein, i.e., the inner mitochondrial membrane. Both with submitochondrial particles from beef heart and especially with mitoplasts from rat liver, incorporation of cholesterol by fusion with sterol-containing liposomes led to a stimulation of ADP-ATP exchange activity, comparable to the effect in the reconstituted system. These results are discussed in relation to the absence of cholesterol in the inner mitochondrial membrane and in the view of the generally accepted ordering effect of cholesterol on phospholipid bilayers. PMID- 6297556 TI - Effect of phosphatidylinositol replacement by diacylglycerol on various physical properties of artificial membranes with respect to the role of phosphatidylinositol response. AB - In an attempt to gain insight into the physiological role of phosphatidylinositol turnover enhanced by extracellular stimuli, the physical properties of artificial membranes (egg yolk phosphatidylcholine/bovine brain phosphatidylserine) containing phosphatidylinositol or diacylglycerol were studied by ESR using spin probes and freeze-fracture electron microscopy. Diacylglycerol lost both the ability to form lipid bilayer structures and its susceptibility to calcium ions. Yeast phosphatidylinositol included in dipalmitoylphosphatidylcholine liposomes lowered the phase transition temperature of dipalmitoylphosphatidylcholine and expanded the temperature range of phase transition. However, diacylglycerol at the same concentration did not undergo the effects caused by phosphatidylinositol but the phase transition temperature was slightly raised. Phase separation of phosphatidylserine induced by calcium ions was enhanced when the phosphatidylinositol was replaced by diacylglycerol in phosphatidylcholine/phosphatidylserine/phosphatidylinositol (3:5:2, by molar ratio) mixtures. The mobility of phosphatidylcholine spin probe was decreased in phosphatidylcholine/phosphatidylserine/diacylglycerol (3:5:2, by molar ratio) liposomes compared with phosphatidylcholine/phosphatidylserine/phosphatidylinositol (3:5:2, by molar ratio) liposomes. An additional component from protonated stearic acid spin probes was observed in phosphatidylcholine/phosphatidylinositol (8:2, by molar ratio) liposomes at 40 degrees C, whereas the component was not seen in phosphatidylcholine/diacylglycerol (8:2, by molar ratio) liposomes. This may indicate the alteration of surface charge induced by the replacement of phosphatidylinositol by diacylglycerol. Indeed, in the presence of 1 mM Ca2+, the additional component was removed by an electrostatic interaction between Ca2+ and phosphatidylinositol molecules in phosphatidylcholine/phosphatidylinositol liposomes at 40 degrees C. These results support the hypothesis that the enhanced turnover of phosphatidylinositol may play a triggering role for various cellular responses to exogenous stimuli by altering membrane physical states. PMID- 6297557 TI - The stimulating effect of 3',5'-(cyclic)adenosine monophosphate and lipolytic hormones on 3-O-methylglucose transport and 45Ca2+ release in adipocytes and skeletal muscle of the rat. AB - (1) In order to assess the possible role of 3',5'-(cyclic)adenosine monophosphate (cAMP) in the control of glucose transport, the effect of the nucleotide or agents known to increase its intracellular concentration on sugar transport or 45Ca2+ washout were characterized in epididymal fat pads, free fat cells and soleus muscles of the rat. (2) When added to the incubation medium, cAMP (0.1-2.0 mM) stimulated 3-O-[14C]methylglucose washout from fat pads. This effect was abolished by cytochalasin B, and additive to that induced by submaximal (10-25 microU/ml), but not by supramaximal (10 microU/ml) concentrations of insulin. (3) cAMP (2 mM) stimulated the conversion of [U-14C]glucose into CO2 and triacylglycerols. This effect was additive to that of insulin (100 microU/ml). (4) ACTH, glucagon, adrenaline, noradrenaline and salbutamol, which are all known to increase the cAMP content of adipose tissue, stimulated the washout of 3-O [14C]methylglucose and 45Ca2+ from preloaded fat pads. The fractional losses of the two isotopes were significantly correlated (P less than 0.001, r = 0.73). (5) In free fat cells, adrenaline (10(-6) M) and salbutamol (10(-5) M) stimulated the uptake of 3-O-[14C]methylglucose, and salbutamol (10(-5) M) did not interfere with the stimulating effect of insulin (25 microU/ml) on sugar uptake. (6) In rat soleus muscles, adrenaline and salbutamol produced a dose-dependent stimulation of the washout of 3-O-[14C]methylglucose and 45Ca2+. The effect of adrenaline on sugar efflux was abolished by propranolol. (7) It is concluded that the activation of the glucose transport system by insulin is unlikely to be mediated by a drop in the cellular concentration of cAMP. An increase in cAMP brought about by beta-adrenoceptor agonists or lipolytic hormones may induce a mobilization of calcium ions from cellular pools into the cytoplasm, which in turn leads to the activation of the glucose transport system demonstrated in the present as well as in several earlier studies. PMID- 6297558 TI - Headgroup behaviour of an uncharged complex glycolipid. AB - A globoside spin labelled on the terminal sugar residue has been synthesized, and employed in model membranes to study headgroup behaviour of complex uncharged glycolipids. The labelled headgroup demonstrated a high degree of motional freedom limited to the aqueous region of the interface between lipid bilayer and surrounding medium. This observation was unaltered by the presence of a dense, tightly-bound surface layer of peripheral proteins or polysaccharide--which might be expected to reproduce conditions present at a cell surface. Headgroup dynamics were only very modestly correlated with the physical state (i.e., fluidity) of the membrane itself. In spite of the absence of charged sugar residues in globoside, the aspects of its headgroup behaviour monitored here we found to be similar to those of oligosaccharide chains on gangliosides and several sialic acid-rich glycoproteins. PMID- 6297559 TI - Calorimetric investigation of polymyxin binding to phosphatidic acid bilayers. AB - The cooperative binding process between the antibiotic peptide polymyxin-B and negatively-charged phosphatidic acid bilayers was investigated by differential thermal analysis and completed by fluorescence polarization measurements. The sigmoidal binding curves were analyzed in terms of the interaction energy within a domain formed by polymyxin and phosphatidic acid molecules. The formation of such a heterogeneous domain structure was favoured by high concentration of external monovalent ions. The cooperativity of the binding increased while a charge-induced decrease in the phase transition temperature of the pure lipid phase was observed with increasing ion concentration at a given pH. The reduced lateral coupling within the lipid bilayer in the presence of salt ions, as demonstrated by an increase in the lipid phase transition enthalpy, was considered to facilitate the cooperative domain formation. Moreover, an increase in the cooperativity of the polymyxin binding could be observed if phosphatidic acids of smaller chain length and thus of a lowered phase transition temperature were used. By the use of chemically-modified polymyxin we were able to demonstrate the effect of electrostatic and hydrophobic interaction. Acetylated polymyxin with a reduced positive charge was used to demonstrate the pure hydrophobic effect of polymyxin binding leading to a decrease in the phosphatidic acid phase transition temperature by about 20 degrees C. The cooperativity of the binding was strongly reduced. Cleavage of the hydrophobic polymyxin tail yielded a colistinnonapeptide which caused an electrostatically-induced increase in the phosphatidic acid phase transition temperature. With unmodified polymyxin we observed the combined effects of electrostatic as well as hydrophobic interaction making this model system interesting for the understanding of lipid-protein interactions. Evidence is presented that the formation of the polymyxin phosphatidic acid complex is a lateral phase separation phenomenon. PMID- 6297560 TI - Fluorescent labeling of (Na+ + K+)-ATPase by 5-iodoacetamidofluorescein. AB - 5-Iodoacetamidofluorescein (5-IAF) covalently labels dog kidney (Na+ + K+)-ATPase with approximately 2 moles incorporated per mole of enzyme. ATPase and K+ phosphatase activities are fully retained after reaction, and the kinetic parameters for Na+, K+, Mg2+, ATP and p-nitrophenyl phosphate are likewise not significantly affected. The fluorescence of the bound 5-IAF is increased by ATP, Na+, and Mg2+, and decreased by K+. These fluorescence changes likely reflect ligand-induced stabilization of the E1 or E2 states of the enzyme. PMID- 6297561 TI - Bretylium opens mucosal amiloride-sensitive sodium channels. AB - Addition of the quanternary ammonium compound, bretylium, to the outer surface of a frog skin leads to an increase in the potential difference and in the short circuit current across the skin. Bretylium does not have any effect when applied to the inside face of the frog skin. The effect of bretylium is dependent upon the presence of sodium ions in the outer medium; it is depressed when sodium is replaced by choline or potassium but not when lithium substitutes for sodium. The bretylium effect is blocked by the specific sodium channel blocker, amiloride. It is proposed that bretylium opens mucosal, amiloride-sensitive sodium channels. PMID- 6297562 TI - Differential inhibition of a restriction enzyme by quinoxaline antibiotics. AB - The inhibition of cleavage by HpaI at two well-defined restriction sites in linearised phi X174-RF DNA by quinoxaline antibiotics has been investigated. Echinomycin, which displays a certain preference for binding to GC basepairs, inhibits cleavage at one site much more than the other, whereas triostin A, which displays less pronounced sequence-selectivity, inhibits both sites about equally. Other congeners inhibit reaction at the two sites with varying effectiveness. The results demonstrate the usefulness of studying inhibition of cleavage at specific sites by restriction enzymes as a means of exploring the specificity of DNA ligand interactions. PMID- 6297563 TI - Changes in DNA binding by purified simian RNA polymerase II under transcribing and nontranscribing conditions. AB - The interaction of RNA polymerase II from African Green Monkey liver tissue with SV40 DNA was examined by a DNAase protection technique. In the absence of nucleoside triphosphates, simian polymerase binds to nicked, linear SV40 DNA and protects 30 bp of binary complex DNA from DNAase I digestion. With the addition of nucleoside triphosphates to initiate transcription, polymerase protects 40 bp of the ternary complex DNA from DNAase I. Thus, a conformational change in either the polymerase, the DNA, or both occurs during the transition from binary to ternary complex, and this altered conformation allows a larger protection of template DNA. Similar results were seen with Escherichia coli RNA polymerase holoenzyme on SV40 DNA. PMID- 6297564 TI - Characterization of DNA methylation in the rat. AB - In the rat, differentiation and cell proliferation both affect DNA methylation. We studied 5-methylcytosine at the inner cytosine of the sequence C-C-G-G, a common methylation site, using endonuclease MspI (which cleaves C-C-G-G- and C-mC G-G), and its isoschizomer HpaII (which cleaves only C-C-G-G). DNA from all tissues and cell lines studied was methylated at C-C-G-G, at levels ranging from 45 to 80%, but the methylation sites were not distributed uniformly. Our analysis suggests a model in which cells contain variable amounts of three DNA methylation states, averaging 30-40, 70-80 and 95-100% methylation, respectively. One biological parameter that alters methylation is the proliferative state of the cell. We observed that NRK, a non-transformed cell line, increased its DNA methylation from 45 to 67% when monolayer cultures became confluent and non dividing. We also observed that a class of repetitive DNA was completely methylated in DNA from all sources except a transformed cell line. PMID- 6297565 TI - 5.3 S RNA is a discrete cleavage product from the 5'-terminus of 18 S RNA of rat liver ribosomes. PMID- 6297566 TI - Down regulation of enkephalin (delta) receptors. Demonstration in membrane-bound and solubilized receptors. AB - The pentapeptide leucine enkephalin induced down-regulation of enkephalin receptors in neuroblastoma-glioma NG108-15 hybrid cells in a reversible fashion, whereas the stable enkephalin analogue D-Ala2-Met-enkephalinamide (AMEA), and the potent opiate alkaloid, etorphine, had a prolonged effect. The opiate alkaloid, morphine, which has low affinity to delta-type enkephalin receptors of these cells did not induce down-regulation, whereas AMEA decreased the binding of both opiate agonists and antagonists but had no effect on the binding of the alpha 2 adrenergic ligand, [3H]yohimbine. From several experiments that were designed to remove the tightly bound AMEA, and from experiments with solubilized receptor we ruled out the possibility that the decreased binding capacity of enkephalin treated cells reflects only receptor masking. The study suggests that down regulation of enkephalin receptors that may also occur in vivo can account for some of the abnormal physiological responses of subjects treated chromically with opiates. However, since opiates from the morphine type can induce opiate tolerance in vivo, but not down-regulation of enkephalin receptors in the cultured cells, we suggest that down-regulation of delta-type opiate receptors may not be prerequisite for the development of the physiological tolerance/dependence on these alkaloids. PMID- 6297567 TI - The interaction between heme and protein in cytochrome c1. AB - The optical spectrum of reduced bovine cytochrome c1 at 77 K shows a fine splitting of the beta-band, which is indicative of the native conformation of the protein. At room temperature, this conformation is reflected in an absorbance band at 530 nm. The exposure of the heme of ferrocytochrome c1, investigated by means of solvent-perturbation spectroscopy, appears to be extremely sensitive to temperature and SH reagents bound to the oxidized protein. Addition of combinations of potential ligands to the isolated tryptic heme peptide of cytochrome c1 reveals that only a mixture of methionine and cysteine (or their equivalents) generates a beta-band at 77 K which is identical in shape to that of native cytochrome c1. In the EPR spectrum of a complex of ferrocytochrome c1 and nitric oxide at pH 10.5, no hyperfine splitting derived from a second ligated nitrogen atom could be detected. The results indicate that methionine and cysteine are the axial ligands of heme in cytochrome c1. The EPR spectrum of isolated ferricytochrome c1 is that of a low-spin heme iron compound with a gz value of 3.36 and a gy value of 2.04. PMID- 6297568 TI - Cytochrome c oxidase. Time dependence of optical and EPR spectral changes related to the 'oxygen-pulsed' form. PMID- 6297569 TI - Impairment of liver regeneration during inhibition of mitochondrial protein synthesis by oxytetracycline. AB - Under standard conditions, liver regeneration is impaired if mitochondrial protein synthesis is completely blocked. By treating rats with oxytetracycline for various periods of time directly prior to partial hepatectomy, livers were led to a condition of relative deficiency in cytochrome c oxidase and ATP synthetase. To this end, oxytetracycline was administered by means of continuous intravenous infusion up to concentrations of 20 micrograms/ml serum, giving a gradual decrease in cytochrome c oxidase activity. This activity was used as a marker for functionally capable mitochondria and as a tool to monitor the efficiency of inhibition of mitochondrial protein synthesis. It is shown that liver regeneration is strongly impaired after a period of pretreatment of 22 days or more and continuation of oxytetracycline treatment during regeneration. The mitochondrial respiratory capacity is reduced to 14% of the control value under these conditions. To obtain inhibitory levels within the regenerating liver, it was necessary to raise the serum levels slightly above 20 micrograms/ml. This measure is most likely required because of the poor vascularization of the regenerating liver. The serum levels were kept, however, far below those known to inhibit cytoplasmic protein synthesis. The results show that in normal liver the respiratory capacity must be reduced drastically before energy-requiring processes become affected. In Zajdela hepatoma cells, similar effects are found after reduction of the cytochrome c oxidase activity to 38%. This difference in sensitivity is probably based on the different mitochondrial content of liver cells and the liver-derived Zajdela cells. PMID- 6297570 TI - Imidazole chloride and tris-chloride substitute for sodium chloride in inducing high-affinity AdoPP[NH]P binding to (Na+ + K+)-ATPase. AB - Optimal binding of [2,8-3H]AdoPP[NH]P to (Na+ + K+)-ATPase requires 25 mM Na+ (Cl ), 50 mM imidazole+ (Cl-) or 50 mM Tris+ (Cl-). Chloride is essential as counterion. We conclude that imidazole+ and Tris+ are able to bind to the Na+ site, and recommend the use of dilute buffers for studying the partial reactions of (Na+ + K+)-ATPase. In NaCl or the substituting buffers the dissociation constant for the enzyme-AdoPP[NH]P complex at 0 degrees C and pH 7.25 is 0.4 microM, whereas in millimolar MgCl2 it is about 2 microM. These distinct levels in affinity with MgCl2 as compared to NaCl, together with the MgCl2-dependence of photolabelling of the enzyme with ATP analogues (Rempeters, G. and Schoner, W. (1981) Eur. J. Biochem. 121, 131-137), suggest significant changes within the substrate site of (Na+ + K+)-ATPase upon binding of Mg2+ (Cl-)2. PMID- 6297571 TI - Dietary lipid modulation of rat liver mitochondrial succinate: cytochrome c reductase. AB - Diets supplemented with high levels of either saturated fatty acids or unsaturated fatty acids were fed to adult rats for a period of 9 weeks and changes in the liver mitochondrial membrane phospholipid fatty acid composition and thermal behaviour of succinate: cytochrome c reductase were determined. The dietary treatment induced a change in the omega 6 to omega 3 unsaturated fatty acid ratio in the membrane lipids, with the ratio being highest with the unsaturated fatty acid and lowest with the saturated fatty acid diet. Arrhenius plots of succinate: cytochrome c reductase activity exhibited differences in both critical temperature (Tf) and Arrhenius activation energy (Ea) depending on the type of dietary treatment. The Tf was elevated from 23 degrees C in control to 32 degrees C in the saturated fatty acid-supplemented group. No significant effect on the Tf was observed in the unsaturated fatty acid-supplemented group however higher Ea values were observed due to the unsaturated fatty acid diet. The changes in succinate: cytochrome c reductase are probably due to changes in the lipid-protein interactions in the membrane, induced by the dietary lipid supplementation. PMID- 6297572 TI - Alteration of the conductance of Na+ channels in the nodal membrane of frog nerve by holding potential and tetrodotoxin. AB - (1) Na+ currents and Na+-current fluctuations were measured in myelinated frog nerve fibres at 15 degrees C during 7.7 ms depolarizations to V = 40, 60 and 80 mV. (2) The conductance gamma of a single Na+ channel and the number No of channels per node were calculated from ensemble average values of the mean Na+ current and the variance of Na+-current fluctuations. (3) For a hyperpolarizing holding potential of VH = -28 mV the mean values of the channel conductance and number were gamma = 9.8 pS and No = 74000. (4) After changing the holding potential to the resting potential (VH = 0) the conductance gamma increased by a factor of 1.37 whereas the number No decreased by a factor of 0.60. (5) Addition of 8 nM tetrodotoxin at a holding potential of VH = -28 mV increased gamma by a factor of 1.55 and reduced No by a factor of 0.25. (6) The increase of the channel conductance at reduced channel numbers suggests negative cooperativity between Na+ channels in the nodal membrane. PMID- 6297573 TI - Development of the gastrointestinal mucosal barrier. Evidence for structural differences in microvillus membranes from newborn and adult rabbits. AB - Electron spin resonance (ESR) and spin label methods with 5-doxylstearic acid as a probe were used to investigate the structure of microvillus membrane from the small intestine of adult and newborn rabbits. The spin label in microvillus membrane of newborns appeared to be in a more disordered environment than spin label in microvillus membrane of adult animals in the temperature range from 4 to 56 degrees C. In addition, a temperature transition at 39.6 +/- 0.3 degrees C was observed in the temperature dependence of the hyperfine splitting parameter for microvillus membrane from adult animals whereas a linear temperature dependence of the hyperfine splitting parameter was found for microvillus membrane from newborns. Cholera toxin was used as an external stimulus to test for the structural response in these two membrane preparations. Cholera toxin at 6 pM caused a decrease in the hyperfine splitting parameter at temperatures below 40 degrees C and a shift in the temperature break from 39.6 degrees C to 30.7 degrees C in microvillus membrane from adults. Using microvillus membrane from newborns, the temperature dependence of the hyperfine splitting parameter remained linear with a cholera toxin stimulus and the disordering effect of cholera toxin was only observed below 30 degrees C. These studies suggested that microvillus membrane from newborns were inherently more disordered than microvillus membrane from adult animals and that this difference in membrane organization might in part account for the increased attachment and penetration of macromolecules noted during the perinatal period. PMID- 6297574 TI - Cholesterol modulation of (Na+ + K+)-ATPase ATP hydrolyzing activity in the human erythrocyte. AB - The cholesterol content of human erythrocyte membranes has been modified by incubation of intact cells with sonicated egg phosphatidylcholine/cholesterol vesicles and with egg phosphatidylcholine vesicles. (Na+ + K+)-ATPase ATP hydrolyzing activity was measured as a function of membrane cholesterol content. High membrane cholesterol inhibits the ATPase activity of the enzyme and low membrane cholesterol activates that enzyme activity. The most likely mechanism of inhibition is suggested to comprise direct cholesterol-protein interactions which lead to a low activity conformation. Ouabain binding studies show that the inhibition is not due to a loss of enzyme from the membrane. PMID- 6297575 TI - Kinetic studies on the (Na+ + K+)-dependent ATPase evidence for coexisting sites for Na+, K+ and Mg2+. AB - Na+-ATPase activity of a dog kidney (Na+ + K+)-ATPase enzyme preparation was inhibited by a high concentration of NaCl (100 mM) in the presence of 30 microM ATP and 50 microM MgCl2, but stimulated by 100 mM NaCl in the presence of 30 microM ATP and 3 mM MgCl2. The K0.5 for the effect of MgCl2 was near 0.5 mM. Treatment of the enzyme with the organic mercurial thimerosal had little effect on Na+ -ATPase activity with 10 mM NaCl but lessened inhibition by 100 mM NaCl in the presence of 50 microM MgCl2. Similar thimerosal treatment reduced (Na+ + K+) ATPase activity by half but did not appreciably affect the K0.5 for activation by either Na+ or K+, although it reduced inhibition by high Na+ concentrations. These data are interpreted in terms of two classes of extracellularly-available low-affinity sites for Na+: Na+-discharge sites at which Na+-binding can drive E2 P back to E1-P, thereby inhibiting Na+-ATPase activity, and sites activating E2-P hydrolysis and thereby stimulating Na+-ATPase activity, corresponding to the K+ acceptance sites. Since these two classes of sites cannot be identical, the data favor co-existing Na+-discharge and K+-acceptance sites. Mg2+ may stimulate Na+ ATPase activity by favoring E2-P over E1-P, through occupying intracellular sites distinct from the phosphorylation site or Na+-acceptance sites, perhaps at a coexisting low-affinity substrate site. Among other effects, thimerosal treatment appears to stimulate the Na+-ATPase reaction and lessen Na+-inhibition of the (Na+ + K+)-ATPase reaction by increasing the efficacy of Na+ in activating E2-P hydrolysis. PMID- 6297576 TI - The K+-induced apparent heterogeneity of high-affinity nucleotide-binding sites in (Na+ + K+)-ATPase can only be due to the oligomeric structure of the enzyme. AB - K+ induces an apparent heterogeneity among an otherwise homogeneous population of nucleotide-binding sites in (Na+ + K+)-ATPase preparations from pig kidney. With the help of ouabain we show that this heterogeneity cannot be due to a mixture of different and independent sites and conclude that each enzyme molecule must contain two nucleotide site-containing units that show interaction. Na+ induces an apparent heterogeneity among an otherwise homogeneous population of ouabain binding sites. The argument is, therefore, extended to include one ouabain site on each of the structural units that bind nucleotide. All these structural units are shown to hydrolyse substrate at identical rates. Using the presently available molecular weight data, it is concluded that the enzyme is composed of two subunits each possessing one nucleotide-binding site, one ouabain-binding site, one alpha-peptide and the capacity for hydrolysing ATP and p-nitrophenyl phosphate. PMID- 6297577 TI - Solubilization of rat kidney benzodiazepine binding sites. AB - The high-affinity binding site for [3H]Ro 5-4864 has been solubilized from rat kidney using 1% Triton X-100. After lowering the concentration of detergent and using a poly(ethylene glycol) gamma-globulin assay, it has been possible to demonstrate solubilization of about 90% of the binding sites. A single soluble class of binding sites with a Kd of 1.8 nM is found. The order of potency of benzodiazepines is identical for the solubilized receptor and the membrane-bound form. Gel filtration revealed a major peak of binding activity with apparent molecular weight of 215000 and a Stokes' radius of 5.03 nm. PMID- 6297578 TI - Calcium transport and Ca2+-ATPase activity in ram spermatozoa plasma membrane vesicles. AB - Plasma membrane vesicles, isolated from ejaculated ram sperm, were found to contain Ca2+-activated Mg2+-ATPase and Ca2+ transport activities. Membrane vesicles that were exposed to oxalate as a Ca2+-trapping agent accumulated Ca2+ in the presence of Mg2+ and ATP. The Vmax for Ca2+ uptake was 33 nmol/mg protein per h, and the Km values for Ca2+ and ATP were 2.5 microM and 45 microM, respectively. 1 microM of the Ca2+ ionophore A23187, added initially, completely inhibited net Ca2+ uptake and, if added later, caused the release of Ca2+ previously accumulated. A Ca2+-activated ATPase was present in the same membrane vesicles which had a Vmax of 1.5 mumol/mg protein per h at free Ca2+ concentration of 10 microM. This Ca2+-ATPase had Km values of 4.5 microM and 110 microM for Ca2+ and ATP, respectively. This kinetic parameter was similar to that observed for uptake of Ca2+ by the vesicles. The Ca2+-ATPase activity was insensitive to ouabain. Both Ca2+ transport and Ca2+-ATPase activity were inhibited by the flavonoid quercetin. Thus, ram spermatozoa plasma membranes have both a Ca2+ transport activity and a Ca2+-stimulated ATPase activity with similar substrate affinities and specificities and similar sensitivity to quercetin. PMID- 6297579 TI - Cationic polypeptide-induced fusion of acidic liposomes. AB - Fusion of acidic liposomes was induced by Mg2+, Ca2+, polylysine and polymyxin B. The extent of fusion and the concomitant change in liposome permeability induced by divalent cations depended on the concentration of liposomes in the suspension as well as on the cation concentration. In contradistinction, the extent of fusion and the change in permeability induced by the polypeptides depended only on the polycation concentration. The difference in the pattern of interaction, between the liposomes and the various cations, is a result of different binding affinities. The binding of the polypeptides to the liposomes, in contrast to divalent cations, is practically irreversible. The potential of polylysine to induce fusion of acidic phosphatidyl-ethanolamine-devoid liposomes was used to demonstrate that in order to obtain fusion, both membranes involved must be susceptible, at least to a certain degree, to fusion by the proper inducer. When lysophosphatidylcholine substituted for phosphatidylcholine in phosphatidylethanolamine-rich acidic liposomes, extensive polylysine-induced fusion was obtained without concomitant spillage of the liposome contents. PMID- 6297580 TI - Effects of insulin upon ion transport. PMID- 6297581 TI - The membrane channel-forming bacteriocidal protein, colicin El. PMID- 6297583 TI - Dissociation of the Pf1 nucleoprotein assembly complex and characterisation of the DNA binding protein. AB - During replication of bacteriophage Pf1, progeny viral strands are complexed with a single-stranded DNA binding protein, analogous to the gene 5 protein of bacteriophage fd. Using fluorescence spectroscopy, ultracentrifugation and DNA cellulose chromatography, conditions for dissociation of the nucleoprotein have been investigated. The Pf1 protein is unusual in that it is not released from the DNA by 2 M NaCl. Complete separation occurs in 0.6-1.0 M MgCl2, leading to a procedure for the purification of the protein. Two subfractions of the protein can be isolated of isoelectric points 5.9 and 6.4. The molecular weight of the native DNA binding protein has been studied by gel filtration and sedimentation. The major species in solution has a sedimentation coefficient of 2.3 S and a diffusion coefficient of 7.8 X 10(-7) cm2 . s-1, corresponding to a protein dimer (Mr = 30 800). Protein tetramers are induced in the presence of octanucleotides, but not tetranucleotides. Analysis of the ultraviolet spectra of the DNA binding protein and the native nucleoprotein complex indicates a stoichiometry of 3.9 +/- 0.4 nucleotides per protein subunit. The molar extinction coefficient of the DNA when bound to the protein (epsilon 260 = 8100) suggests that the binding protein maintains the DNA in an extended (unstacked) conformation similar to that found in the mature Pf1 virion. PMID- 6297582 TI - Identification of two new promoters probably involved in the transcription of a ribosomal RNA gene of Escherichia coli. AB - The DNA sequence in the region preceding the rrnB gene of Escherichia coli was determined up to the 1821st nucleotide upstream from the beginning of the sequence coding for mature 16 S rRNA. In vitro transcription experiments indicated the presence of two new promoters in this region, located more than 1 kb upstream from the known P1 and P2 promoters of rrnB. Previous electron microscopic studies demonstrated that these sites bind RNA-polymerase very strongly. In vitro transcription, starting at these sites reads through the entire region into the rrnB gene without termination. A similar uninterrupted transcription into rrnB in vivo can be demonstrated by S1-mapping, and by fusing the DNA containing the new promoters (but not P1 and P2) to the lacZ gene. Thus it seems likely that these promoters (P3 and P4) belong functionally to the rrnB gene and play some role in its regulation of expression. PMID- 6297584 TI - Kinetics of yeast cytochrome c peroxidase compound I formation with modified substrates (peroxybenzoic acids). AB - The kinetics of formation of Compound I of yeast cytochrome c peroxidase (ferrocytochrome c:hydrogen-peroxide oxidoreductase, EC 1.11.1.5) with a series of peroxybenzoic acids were studied. Reactivity is affected not only by protein ionization, as in the reaction with H2O2, but also by substrate ionization. The reactivity of negatively charged substrates is markedly lower than that of uncharged species, implying that electrostatic factors profoundly influence substrate binding. The rate constants for neutral peroxybenzoic acids carrying electron-withdrawing substituents increase with increasing peroxy acid pKa. This behaviour suggests that, as previously discussed for reactions of turnip peroxidases, formation of peroxy anion by ionization of substrate within the active site is kinetically important. The results support the mechanism of cytochrome c peroxidase Compound I formation which has been proposed by Poulos and Kraut (J. Biol. Chem. 225 (1980), 8199-8205) on the basis of enzyme structural studies. PMID- 6297585 TI - Fragments of human growth hormone produced by digestion with bromelain. Chemistry and biological properties. PMID- 6297586 TI - Partial characterization of the inactive mutant form of human red cell bisphosphoglyceromutase and comparison with an alkylated form. AB - The trifunctional enzyme bisphosphoglyceromutase (or diphosphoglycerate mutase) (EC 2.7.5.4) was purified from human red cells and injected into two chickens. Specific anti-bisphosphoglyceromutase antibodies were produced that displayed a single precipitation line on Ouchterlony plates and on immunoelectrophoresis. No cross-reaction of these antibodies was detected with phosphoglyceromutase, the common glycolytic enzyme. Immunoneutralization of bisphosphoglyceromutase and of its two other activities, i.e., bisphosphoglycerate phosphatase and phosphoglyceromutase, was observed for a purified preparation. The anti bisphosphoglyceromutase antibody reacts with the inactive enzyme present in the hemolysate of a mutant human subject. It also binds bisphosphoglyceromutase inactivated by N-ethylmaleimide, a strong alkylating agent of SH groups. Active bisphosphoglyceromutase is stable at 55 degrees C, whereas the inactive forms of the mutant and of the alkylated hemolysates are thermolabile. These forms can be protected against thermal precipitation by 4 mM 2,3-diphosphoglycerate and 4 mM 3 phosphoglycerate. These findings afford evidence that the binding of the substrates on the bisphosphoglyceromutase molecule is not prevented by alkylation nor by the mutation of the hereditary inactive enzyme. PMID- 6297587 TI - Synthesis and properties of spin-labeled angiotensin derivatives. PMID- 6297588 TI - Sequential hydrolysis of the gamma- and beta-phosphate groups of ATP by the ATP diphosphohydrolase from pig pancreas. AB - The ATP diphosphohydrolase (EC 3.6.1.5) from pig pancreas hydrolyzes triphospho- and diphosphonucleosides. The reaction products of ATP hydrolysis are ADP, AMP and orthophosphate, but AMP accumulates at a faster rate than ADP. A time-course study showed a simultaneous breakdown of ATP and ADP with initial rates for ATP and ADP hydrolysis of 2.1 and 3.8 mumol/min per mg protein, respectively. However, the rates reached similar values toward the end of the incubation period. According to double reciprocal plots and Dixon plots, the Km values for ATP and ADP are similar, Vmax for ADP hydrolysis is twice the Vmax for ATP hydrolysis and both nucleotides are competitive inhibitors of the other with their Ki values similar to their Km. These results are consistent with a sequential hydrolysis of the two diphosphoester bonds of ATP: ATP first binds to the enzyme, its gamma-phosphate group is hydrolyzed and released, resulting in an enzyme-ADP complex which either breaks down to free enzyme and ADP or is further processed via hydrolysis of the beta-phosphate group, releasing free enzyme, AMP and Pi. The experimental data showed that the processing step is favored. PMID- 6297589 TI - The binding of bilirubin to albumin. A study using spin-labelled bilirubin. AB - Binding between human serum albumin and a spin-labelled derivative of bilirubin was investigated by circular dichroism, fluorescence quenching, electron spin resonance and visible spectroscopy. The orders of magnitude of the binding constants obtained by flurorescence quenching and electron spin resonance spectroscopies were 10(7) and 10(3) 1 . mol-1, respectively. These data suggest that most spin-labelled bilirubin interacts with human serum albumin at the side not holding the spin-labelled side-arm. CD measurements showed the presence of at least two sites, associated with opposite Cotton effects. It is worthy of note that the Cotton sign of the first site is inverted with respect to the corresponding one of bilirubin. CD measurements on mixed systems (spin-labelled bilirubin/human serum albumin/bilirubin) were also performed. The decomposition of the ternary curves shows that the rotatory power of bilirubin bound to human serum albumin is higher in the ternary system than in the binary (bilirubin/human serum albumin). The corresponding CD measurements for the binding between spin labelled bilirubin and bovine serum albumin are also reported and discussed. PMID- 6297590 TI - Tryptic digest of the alpha subunit of lamb kidney (Na+ + K+)-ATPase. AB - The Mr approximately equal to 100 000 alpha subunit was prepared from highly purified lamb kidney (Na+ + K+)-ATPase. Its N-terminal sequence is Gly-Arg-Asx Lys-Tyr-Glu. The alpha subunit was S-carboxymethylated, succinylated, and cleaved at its 40 arginine residues with trypsin. Four major, well-differentiated peptide fractions (A to D) were obtained by chromatography of the digest on a Sephadex G 50 column. Fraction A eluted at the void volume of the column and contained aggregated, very hydrophobic peptides, possibly from regions of alpha that are buried within the membrane lipid bilayer in the native enzyme. Fractions B to D, which together accounted for about 75% of the total protein, contained water soluble peptides. To test the feasibility of using antibodies to identify and purify specific peptides of alpha subunit, studies were carried out using antibodies to native (Na+ + K+)-ATPase. Carboxymethylation and succinylation did not significantly decrease total antibody binding to alpha subunit, although the affinity of the anti-(Na+ + K+)-ATPase antibodies for alpha subunit was reduced by about 50%. The tryptic peptides of alpha subunit also retain significant immunochemical reactivity. Fractions A, B and C (but not D) of the digest all bind antibodies. To characterize further the tryptic digest, 16 peptides from fraction D were isolated and sequence studies on these were carried out. PMID- 6297591 TI - Temperature dependence of the electronic spin-lattice relaxation time in a 2-iron 2-sulphur model complex. AB - The complex [Fe2S2(S2-o-xylyl)2]2- in DMF (dimethylformamide), DMSO (dimethylsulphoxide) or a 1:1 DMF/DMSO mixture, a model for the chromophore in the 2Fe-2S proteins (ferredoxins), has been reduced and studied by conventional EPR over a temperature range. The low-field feature of the spectrum, Hz, has been computer simulated in order to analyse the lineshape in terms of a convolution product of Lorentzian and Gaussian distributions. The Gaussian contribution to the linewidth and a fixed part of the Lorentzian contribution, which is a function of the solvent and the way it freezes, were measured at a low temperature (less than or equal to 100 K) and subtracted from the linewidths in the higher-temperature range (130-200 K). The variable Lorentzian contribution thus obtained was related to spin-lattice relaxation times. The spin-lattice relaxation times of the sample having 1:1 DMSO/DMF solvent were measured in the range 6 to 11 K by the saturating pulse technique and in the range 10 to 65 K by continuous saturation methods. Up to 65 K the results follow the law 1/T1 alpha T4.5, a relationship which is not readily interpreted in terms of a simple Debye model. At higher temperatures the results may be interpreted in terms either of a dominant Orbach mechanism involving excited states at approx. 900 +/- 50 cm-1 (DMSO, DMF) or 770 +/- 50 cm-1 (1:1 DMSO/DMF), or of a Raman process in which 1/T1 alpha T7.5. The former is compatible with the two-phonon process already described in some ferredoxins, especially those with little anisotropy (gy - gx approximately 0.0) which have characteristically high [J] values. PMID- 6297592 TI - Cooperative binding of hexadecyltrimethylammonium chloride and sodium dodecyl sulfate to cytochrome c and the resultant change in protein conformation. AB - The binding of hexadecyltrimethylammonium chloride (HTAC) and sodium dodecyl sulfate (SDS) to cytochrome c was determined by potentiometric titration and the corresponding changes in protein conformation by circular dichroism (CD). The binding isotherms were biphasic; about 20 surfactant cations or anions were bound to cytochrome c in the first phase. Another 30 or so HTA+ ions were bound in the second phase, which was below the critical micelle concentration of the surfactant, but the binding of dodecyl sulfate ions in the second phase increased sharply near the critical micelle concentration. The binding of both surfactants was highly cooperative and was endothermic; the data in the first phase fitted the Hill plot. The corresponding change in the secondary structure of cytochrome c was small; the CD spectra in the ultraviolet region showed a moderate increase in the helicity in HTAC solution and some changes in conformation in SDS solution. However, the CD spectra for the Soret band indicated a marked change in the local conformation around the heme. PMID- 6297593 TI - Deuterium exchangeable proton hyperfine resonances of low-spin cytochrome c peroxidase and the mechanism of peroxidase catalysis. AB - Deuterium exchangeable hyperfine proton NMR resonances of cytochrome c peroxidase (EC 1.11.1.5) are identified in H2O solutions of the enzyme. One of these is assigned to the proximal histidine's imidazole N-H. Its shift and pH dependence indicate that an imidazolate form, which has been postulated for peroxidases, is ruled out for cytochrome c peroxidase-cyanide. A qualitative comparison of relative heme-pocket dynamics is also possible. When the bulk water resonance is irradiated with a continuous, but off acquisition, decoupler frequency the N-H resonance shows no intensity loss, indicating that saturation transfer between the proximal histidine and solvent water is either minimal, or extremely slow. PMID- 6297594 TI - Purification and characterization of phosphopentomutase from rat liver. PMID- 6297595 TI - Properties and function of the two hemes in Pseudomonas cytochrome c peroxidase. AB - The oxidation-reduction potentials of the two c-type hemes of Pseudomonas aeruginosa cytochrome c peroxidase (ferrocytochrome c:hydrogen-peroxide oxidoreductase EC 1.11.1.5) have been determined and found to be widely different, about +320 and -330 mV, respectively. The EPR spectrum at temperatures below 77 K reveals only low-spin signals (gz 3.24 and 2.93), whereas optical spectra at room temperature indicate the presence of one high-spin and one low spin heme in the enzyme. Optical absorption spectra of both resting and half reduced enzyme at 77 K lack features of a high-spin compound. It is concluded that the heme ligand arrangement changes on cooling from 298 to 77 K with a concomitant change in the spin state. The active form of the peroxidase is the half-reduced enzyme, in which one heme is in the ferrous and the other in the ferric state (low-spin below 77 K with gz 2.84). Reaction of the half-reduced enzyme with hydrogen peroxide forms Compound I with the hemes predominantly in the ferric (gz 3.15) and the ferryl states. Compound I has a half-life of several seconds and is converted into Compound II apparently having a ferric-ferric structure, characterized by an EPR peak at g 3.6 with unusual temperature and relaxation behavior. Rapid-freeze experiments showed that Compound II is formed in a one-electron reduction of Compound I. The rates of formation of both compounds are consistent with the notion that they are involved in the catalytic cycle. PMID- 6297596 TI - Determination of the coordination geometry at the heme iron in three cytochromes c from Saccharomyces cerevisiae and from Candida krusei based on individual 1H NMR assignments for heme c and the axially coordinated amino acids. AB - The 1H-NMR lines of heme c and the axial ligands in reduced and oxidized Iso-1 and Iso-2 cytochromes c from Saccharomyces cerevisiae and in cytochrome c from Candida krusei were individually assigned and the conformation of the coordination sphere of the heme iron was investigated with the use of proton proton Overhauser enhancement measurements and circular dichroism spectroscopy. The coordination geometry of the axial methionine and the axial histidine and the electronic structure of the heme were found to be closely similar in these yeast cytochromes c and in mammalian cytochromes c. In particular, R chirality at the sulfur atom of the iron-bound methionine was observed in both groups of proteins. Additional nuclear Overhauser enhancement studies of the spatial arrangement relative to the heme group of amino acid side-chains in the heme crevice of yeast ferrocytochromes c showed that the conformational homologies extend beyond the immediate coordination sphere of the heme iron. These data provide a conformational basis for observations on the functional properties of cytochromes c from yeast and mammalian species, which were reported previously by other groups. PMID- 6297597 TI - Individual 1H-NMR assignments for the heme groups and the axially bound amino acids and determination of the coordination geometry at the heme iron in a mixture of two isocytochromes c-551 from Rhodopseudomonas gelatinosa. AB - This paper describes chemical and physicochemical studies of two small isocytochromes c-551 (approx. 9000 dalton) from Rhodopseudomonas gelatinosa. In spite of numerous amino acid substitutions in the N-terminal half of the sequence the two isoproteins could not be separated by the procedures used, presumably because they have identical size, charge and isoelectric points. Individual assignments of the 1H-NMR lines of heme c and the axial ligands to the heme iron were therefore obtained by nuclear Overhauser enhancement measurements and saturation transfer experiments in a mixed solution of the two isocytochromes c 551. The conformation of the coordination sphere was investigated by additional 1H-NMR and circular dichroism studies. For both isoproteins the electronic structure of the heme and the chirality of the methionine attachment to the iron were found to coincide with those in Pseudomonas cytochromes c-551, i.e., S chirality was observed for the axial methionine. The Rps. gelatinosa cytochromes c-551 thus differ from mammalian, yeast, Euglena gracilis and Rhodospirillum rubrum cytochromes c, which all have R chirality at the axial methionine and concomitantly a characteristically different electronic heme structure. This is the first observation of S chirality of the axially bound methionine in a species outside the Pseudomonas family. The redox potentials of the two isocytochromes c 551 of Rps. gelatinosa differ by approx. 120 mV, and there is no cross-exchange of electrons between the two species. The two isoproteins could thus function in two different, parallel electron-transfer chains or at two different locations in a single transfer sequence. PMID- 6297598 TI - Isolation and characterization of a ferredoxin from Mycobacterium smegmatis Takeo. PMID- 6297599 TI - An EPR study of the blue copper center in horse liver alcohol dehydrogenase. AB - Active-site specifically reconstituted Cu2+ horse liver alcohol dehydrogenase (alcohol:NAD+ oxidoreductase, EC 1.1.1.1) shows optical and EPR spectra similar to those of native blue copper (Type 1) proteins. EPR spectra at different temperatures and frequencies reveal a heterogeneity of the copper center: a minor 'non-blue' species with axial line shape (g parallel = 2.16, g perpendicular = 2.04; A parallel = 100 x 10(-4) cm-1), which accounts for approximately 10% of the total copper and is not accessible to ligands and a major blue species with rhombic line shape (g1 = 2.21, g2 = 2.06, g3 = 2.03, A1 = 50 x 10(-4) cm-1, A2 = 30 x 10(-4) cm-1, A3 = 76 x 10(-4) cm-1), which is accessible to ligands and participates in redox reactions. The major blue species in cupric horse liver alcohol dehydrogenase is metastable, since it is reduced in a process markedly accelerated in the presence of oxygen or hydrogen peroxide. In addition, the reduction depends on the presence of exogenous metal ligands or coenzymes. Whereas the binary complex enzyme-NAD+ is even more susceptible to bleaching than the free enzyme, the cupric center is stable towards bleaching in the binary complex enzyme-NADH. In the discussion the redox properties and coordination chemistry of Cu2+ in horse liver alcohol dehydrogenase and copper proteins are compared. PMID- 6297600 TI - Assignment of hyperfine shifted resonances in high-spin forms of cytochrome c peroxidase by reconstitutions with deuterated hemins. AB - Assignments of hyperfine shifted proton resonances for the high-spin forms of cytochrome c peroxidase (EC 1.11.1.5) have been made (cytochrome c peroxidase, cytochrome c peroxidase-F) employing the technique of reconstituting the apoprotein with specifically deuterated protohemin IX derivatives. The results show that the heme methyl group pattern differs significantly from similar assignments made for metmyoglobin. In cytochrome c peroxidase the methyl pattern is 5 greater than 1 greater than 8 greater than 3. For cytochrome c peroxidase-F the pattern is 5 greater than 8 greater than 1 greater than 3, but the resonances are not shifted as far downfield and they exhibit a narrower spread. For myoglobin the relative methyl ordering has previously been shown to be 8 greater than 5 greater than 3 greater than 1. Several conclusions have been reached, including confirmation of the essential correspondence between the solution- and crystal-derived data for several heme crevice structural features. The pH dependence of the cytochrome c peroxidase-F methyl resonances is also presented and is shown to differ from native peroxidase. For cytochrome c peroxidase-F smooth, continuous titrations are observed with no evidence of the second conformation which was found for the native enzyme. PMID- 6297601 TI - Electron paramagnetic resonance studies of highly anisotropic low-spin states of ferrimyoglobin derivatives. AB - The effects of addition of nitrogenous bases, which gave low-spin ferric porphyrin complexes with highly anisotropic g values, were investigated for ferrimyoglobin by low-temperature EPR measurements. Concomitant denaturation of myoglobin upon addition of the exogenous bases was also of interest. By addition of pyridine-type bases under regulated pH, Mb(Fe3+) complexes showing EPR spectra with highly anisotropic g values were formed. These complexes have the electronic states close to the spin-crossover point but not so close as that of the ferric porphyrin highly anisotropic low-spin (HALS) complexes previously reported. Several types of low-spin species, LSi, LSa and LSb, were produced by the denaturation of myoglobin caused by addition of some exogenous ligands. The LSi was assigned to a complex with histidine-E7 coordinated on the sixth position and LSa to the one with OH- and histidine-F8[Im0]. PMID- 6297602 TI - Comparison of human and rat liver microsomes by spin label and biochemical analyses. AB - Detailed lipid analyses of human and rat liver microsomes revealed interesting differences. It was found that human liver microsomes contain twice as much lipid as those from the rat. This increased lipid content is not associated with an increase in content of a particular lipid class; human liver microsomes contain higher amounts of each of the lipid classes. Human and rat liver microsomes differ especially in the essential fatty acid composition of total lipids and phospholipids: human liver microsomes contain more linoleic acid and less arachidonic acid than those of the rat. Such a pattern of distribution of fatty acids is similar to that previously reported for human liver mitochondria and has not been reported for other species. Although the previously reported for human liver mitochondria and has not been reported for other species. Although the unsaturation of lipids is lower in human than in rat liver microsomes, spin label studies revealed a higher fluidity in human membranes. It is suggested that this might arise from a lesser immobilization of lipids by proteins in human liver subcellular membranes. PMID- 6297603 TI - On the suitability of organotypic cultures of fetal rat lung type II cells for biochemical studies concerning development. AB - When organotypic cultures of fetal rat lung epithelial cells are initiated with undifferentiated cells, the cells differentiate into type II cells (Douglas, W.H.J., McAteer, J. A., Smith, J.R. and Braunschweiger, W.R. (1979) Int. Rev. Cytol., Suppl. 10, 45-65). This conclusion was based only on morphologic studies. The present study was undertaken to investigate whether such maturation in culture could also be demonstrated biochemically. In organotypic cultures initiated with epithelial cells from fetal rat lungs at 17-days gestation, the amount of phospholipids increased for at least 10 days. However, no change took place in the percentage of phosphatidylglycerol nor in the ratio of disaturated to total phosphatidylcholine. In cultures initiated with cells obtained at day 17 of gestation the specific activity of cholinephosphate cytidylyltransferase reached a maximum after approximately 3 days, followed by a decrease. A similar profile was obtained, however, if the culture was started at day 20 of gestation. This indicates that the activity profiles obtained in the organotypic cultures reflect changes caused by the culture conditions rather than changes caused by maturation. From these investigations it is concluded that biochemical studies on type II cell development using organotypic cultures as model should be interpreted with caution. PMID- 6297604 TI - Mechanisms involved in the synthesis of disaturated phosphatidylcholine by alveolar type II cells isolated from adult rat lung. AB - 1. Alveolar type II cells isolated from adult rat lung incorporated radioactively labelled palmitate predominantly into the 2-position of disaturated phosphatidylcholine. In disaturated diacylglycerol, however, the radioactivity was almost equally distributed between the 1- and 2-positions. 2. Exposure of isolated type II cells to the phospholipase A2 inhibitors 4-bromophenacylbromide or quinacrine dihydrochloride led to a decreased synthesis of total phosphatidylcholines from various labelled precursors. Interestingly, it also led to an increased degree of unsaturation of the phosphatidylcholines synthesized by the cells. 3. Incubation of type II cell sonicates with radioactively labelled CDPcholine resulted in the formation of labelled phosphatidylcholine; 56% of this phosphatidylcholine appeared to be disaturated. In similar experiments with homogenates from whole lung, 20% of the synthesized phosphatidylcholine was disaturated. 4. These results suggest that both direct synthesis de novo and remodeling of 1-saturated-2-unsaturated phosphatidylcholines contribute to the biosynthesis of disaturated phosphatidylcholine in isolated alveolar type II cells. PMID- 6297605 TI - Phosphatidic acid phosphatase activity in subcellular fractions derived from adult rat type II pneumocytes in primary culture. AB - The development of a method for the subcellular isolation of lamellar body, microsomal, mitochondrial and cytosolic enriched fractions from the adult rat type II pneumocyte is presented. These fractions were used to investigate the distribution and specificity of phosphatidic acid phosphatase (EC 3.1.3.4). The cytosolic enriched fraction possessed the greatest proportion of total phosphatidic acid phosphatase activity and the highest relative specific activity. The small amounts of phosphatase activity in the lamellar body, microsomal and mitochondrial enriched fractions correlated best with a nonspecific phosphatase activity. 1,2-Disaturated phosphatidic acid served as a slightly better substrate than did 1-saturated, 2-unsaturated phosphatidic acid in the cytosolic enriched fraction. This observation indicates that the de novo pathway could contribute to dipalmitoylphosphatidylcholine synthesis in the rat type II pneumocyte. PMID- 6297606 TI - Fatty acyl delta 6 desaturation activity of cultured human endothelial cells. Modulation by fetal bovine serum. AB - Human endothelial cells from umbilical vein actively desaturate [14C]linoleate and synthesize icosatrienoate, arachidonate and docosatetraenoate. Desaturation and chain elongation of 9,12,15-[14 C]linolenate (n - 3) by these cells is more extensive than that of [14 C]linoleate. Both confluent primary monolayers and subconfluent subcultures exhibit greater fatty acyl CoA delta 6-desaturase activity when growth and incubation media contain 2.5% fetal bovine serum instead of 10%. Prior growth with 20% serum diminishes the extent of subsequent linoleate desaturation. Use of medium supplemented with 20-100 microM oleate results in up to 67% inhibition of [14 C]arachidonate synthesis. These results indicate that, despite previously published reports to the contrary, human vascular endothelial cells are similar to other normal mammalian cells in having fatty acyl delta 6 desaturase activity. Suppression of endogenous arachidonate synthesis by elevated levels of serum lipids may impair endothelial cell function. PMID- 6297607 TI - Cyclic AMP-dependent protein phosphorylation and dephosphorylation alter phosphate transport in canine renal brush border vesicles. AB - We have previously demonstrated that the cAMP-dependent phosphorylation of two or more proteins (bands IX and V) in membrane vesicles isolated from the renal brush border from kidneys of dogs, was associated with decreased Na+-dependent Pi transport. In the present studies a specific dephosphorylation of band IX was demonstrated in brush border vesicles incubated in the absence of F- which had been used in previous studies to inhibit phosphoprotein phosphatase activity. Dephosphorylation of band IX was 80% complete after 5 min of incubation at which time inhibition of Pi transport in membrane vesicles which had been phosphorylated in the presence of cAMP could no longer be demonstrated. Dephosphorylation of band IX was no different in vesicles from kidneys originating from parathyroidectomized dogs prior to or following the administration of parathyroid hormone in vivo and normal dogs. We conclude that the cAMP-dependent phosphorylation of brush border membrane proteins may mediate a phosphaturic effect of the hormone. Parathyroid hormone-induced phosphaturia may be terminated through the action of a specific membrane phosphoprotein phosphatase. PMID- 6297608 TI - Stimulation of renal gluconeogenesis by exogenous adenine nucleotides. AB - 1. Tubule fragments were isolated from renal cortex of fed rats. 2. Gluconeogenesis from lactate was significantly increased by low concentrations of exogenous ATP, ADP, AMP adenylyl (beta, gamma-methylene)diphosphonate and, to a lesser extent, by ITP and inosine. GTP was slightly inhibitory. Hypoxanthine was ineffective. Exogenous adenosine deaminase slightly decreased gluconeogenesis and was additive in effect to GTP. Adenosine deaminase did not abolish the stimulatory effects of ATP or cyclic AMP. 3. 40 microM ATP also stimulated gluconeogenesis from pyruvate, malate, succinate, 2-oxoglutarate and glutamine, but had no effect when glycerol or fructose were used as substrates. 4. With lactate as substrate the effect of 40 microM ATP was additive to the maximal stimulations of gluconeogenesis seen with 1 microM noradrenalin or 0.1 microM angiotensin II, but was not additive to the stimulatory effect of 0.1 mM cyclic AMP. 5.40 microM ATP had no effect upon either the tubule content of cyclic AMP or upon 45Ca efflux from prelabelled tubules. 6. Addition of ouabain or removal of extracellular K+ diminished the stimulatory effects of ATP and cyclic AMP. 7. Extracellular ATP was rapidly metabolized by tubule fragments, with resulting accumulation of adenosine. Further metabolism resulting in formation of inosine, hypoxanthine, xanthine and uric acid was also observed. Cyclic AMP was metabolized less rapidly, with no accumulation of adenosine. 8. The effects of purinergic agents on gluconeogenesis are discussed. PMID- 6297609 TI - N-(6-Aminohexyl)-5-chloro-1-naphthalenesulfonamide(W-7), a calmodulin antagonist, also inhibits phospholipid-sensitive calcium-dependent protein kinase. AB - N-(6-Aminohexyl)-5-chloro-1-naphthalenesulfonamide(W-7), commonly regarded as a calmodulin antagonist, inhibited phospholipid-sensitive Ca2+ -dependent protein kinase and to a lesser extent cyclic GMP- and cyclic AMP-dependent protein kinases. Kinetic studies of the inhibition of the homogeneous spleen phospholipid sensitive Ca2+ -dependent protein kinase indicated that W-7 inhibited the enzyme activity competitively with respect to phospholipid (Ki = 60 microM). N-(6 Aminohexyl)-1-naphthalenesulfonamide (W-5) was found to be much less potent than W-7. The findings indicate that W-7 was able to inhibit a variety of protein kinases, in addition to those requiring calmodulin previously reported. PMID- 6297610 TI - Calmodulin stimulates polyamine-responsive protein kinase in the absence of Ca2+. AB - A cyclic nucleotide-independent, polyamine-responsive protein kinase from the cytosol of Morris hepatoma 3924A, which phosphorylated heat-stable endogenous substrates and casein in the presence of polyamines (Criss, W.E., Yamamoto, M., Takai, Y., Nishizuka, Y. and Morris, H.P. (1978) Cancer Res. 38, 3540-3545) was observed to be stimulated by an endogenous protein activator. This protein activator was identified to be calmodulin. the polyamine-responsive protein kinase was also stimulated by purified calmodulin, but only in the presence of polyamines such as polylysine. This action of calmodulin did not require Ca2+ for activation of the enzyme; and activation occurred in the presence of EGTA. DNA and RNA inhibited the polyamine-responsive protein kinase, either in the presence or absence of Ca2+. Purified calmodulin, in the presence of cyclic AMP or cyclic GMP, did not activate the protein kinase. Therefore, polyamines such as polylysine are an absolute requirement for this expression of calmodulin action. The increased enzyme activity by calmodulin was accompanied with an increased Vmax and with no changes in the Km (ATP). High levels of cation, up to 100 mM Mg2+, did not effect the action of calmodulin. These results indicate that tumor cytosolic polyamine-responsive protein kinase is regulated by calmodulin, the latter being increased in the tumor tissue. PMID- 6297611 TI - Temperature dependence of the dissociation rate constants for the nonactivated (molybdate-stabilized) and activated progesterone receptor-hormone complexes from the chick oviduct. AB - Both the nonactivated and activated forms of the chick oviduct cytosol progesterone receptor-hormone complexes displayed first-order dissociation kinetics at temperatures between 0 and 25 degrees C. The rate constant was always 2-3-times greater for the nonactivated than for the activated complex. The thermodynamic parameters calculated from the Eyring plot for the nonactivated and activated forms, respectively, were: delta H+ = 28.6 +/- 0.2 and 29.9 +/- 1.5 kcal/mol; -T delta S+ = 7.4 +/- 0.6 and 7.7 +/- 1.6 kcal/mol; and delta G+ = 21.3 +/- 0.5 and 22.1 +/- 0.1 kcal/mol. These values suggest that activation results in an increase in enthalpy of the ligand-receptor interaction, thus stabilizing the complex. The dissociation rate constants for the native complex obtained by two different experimental approaches, namely, isotope dilution ('chase') and dissociation against charcoal, indicated the absence of cooperativity in the receptor-ligand binding. PMID- 6297612 TI - Studies on the temporal relationships between receptor binding and cellular response, especially concerning the luteinizing hormone release rate. PMID- 6297613 TI - 3':5'-cyclic-nucleotide phosphodiesterase in the bovine pituitary gland. AB - Cyclic-AMP phosphodiesterase activity in the homogenate of the anterior pituitary gland was 2-fold higher than that in the homogenate of the posterior pituitary, whereas cyclic-GMP phosphodiesterase activity was dominant in the posterior homogenate. There were two peaks of cyclic-AMP phosphodiesterase activity with different isoelectric points of 4.3 and 5.2. Fraction I had a molecular weight of 240 000 and a sedimentation coefficient of 6.2 S; fraction II had a molecular weight of 180 000 and a sedimentation coefficient of 3.1 S. Cyclic AMP hydrolytic activity in the supernatant of the posterior lobe corresponded to fraction I in the anterior lobe. Cyclic GMP hydrolytic activity in both the anterior and posterior lobes (activated by Ca2+/calmodulin) had an isoelectric point of 5.2, a molecular weight of 240 000 and a sedimentation coefficient of 6.2 S. Cyclic AMP and GMP hydrolytic activities in both the anterior and posterior lobes appeared in fraction I and did not separate when the preparations were mixed before electric focusing or sucrose density gradient procedures. Cyclic AMP hydrolytic activity in fraction II could be separated from cyclic GMP hydrolytic activity. PMID- 6297614 TI - Collagen, collagenase and collagenolytic activity in rat Graafian follicles during follicular growth and ovulation. AB - Follicles were dissected from the ovaries of immature rats at intervals after subcutaneous injection of 20 IU of pregnant mare's serum gonadotropin. A surge of luteinizing hormone was observed at 54 h and ovulation occurred at 64-66 h. The follicular volume between 36 and 48 h, then doubled again shortly before ovulation. The collagen content of the follicles increased 3-fold from 35 to 56 h, but decreased significantly (25%) from 61 to 66 h. Follicle homogenates, activated with trypsin or aminophenylmercuric acetate, digested Type I collagen at 28 degrees C to produce typical of a true collagenase. Collagenolytic activity assayed against endogenous collagen at 37 degrees C did not change significantly between 38 and 66 h. PMID- 6297615 TI - Studies of albumin binding to rat liver plasma membranes. Implications for the albumin receptor hypothesis. AB - To characterize a previously proposed hepatocyte albumin receptor, we examined the binding of native and defatted 125I-labeled rat albumin to rat liver plasma membranes. After incubation for 30 min, binding was determined from the distribution of radioactivity between membrane pellet and supernatant following initial centrifugation (15000 X g for 15 min), and after repeated cycles of washing with buffer and re-centrifugation. 125I-labeled albumin recovered in the initial membrane pellet averaged only 4% of that incubated. Moreover, this albumin was only loosely associated with the membrane, as indicated by recovery in the pellet of under 0.5% of the counts after three washes. Binding of 125I labeled albumin to the plasma membranes was no greater than to erythrocyte ghosts, was not inhibited by excess unlabeled albumin, and was not decreased by heat denaturation of the membranes, all suggestive of a lack of specific binding. Failure to observe albumin binding to the membranes was not due to a rapid dissociation rate or 'off-time', as incubations in the presence of sufficient ultraviolet light to promote covalent binding of ligands to receptors did not increase 125I counts bound to the membrane. Finally, affinity chromatography over albumin/agarose gel of solubilized membrane proteins provided no evidence of a membrane protein with a high affinity for albumin. These studies, therefore, do not support the hypothesis that liver cell plasma membranes contain a specific albumin receptor. PMID- 6297616 TI - Modulation of the activity of 5'-nucleotidase by the transfer of non-esterified cholesterol to rat-liver microsomal fraction and evidence for regulation of the concentration of non-esterified cholesterol in plasma membranes in vivo. PMID- 6297618 TI - [Microvicosity of water medium in plant objects by a spin probe method]. AB - Correlation times in rotation diffusion of the radical in leaves and roots of wheat "Moskovskaya 35" were found under different conditions from the ESR spectra parameters of iminoxyl radical 2,2,6,6-tetramethyl-4-oxopiperidine-1-oxyl injected into these objects. The radical was shown to be transported along the plant with water current. Change of viscosity in dehydrated plant was studied. Temperature relationship (in the range 10-35 degrees C) of the probe surrounding in wheat leaves was investigated. A change of stabilizing effect of KCl admixture on the short-range order in water structure of the plants as compared to pure water was found. PMID- 6297617 TI - [Conductance of cytoplasmic membrane of photoreceptors by the method of intracellular dialysis]. AB - To clear up the effect of agents proposed as a mediator in the phototransduction process on the rod outer segment cytoplasmic membrane a technique based on the intracellular dialysis method was developed. It was shown that in the presence of nucleoside-triphosphates 3',5'-cGMP increased cytoplasmic membrane sodium conductance of dialyzed rod outer segment by 1-4 nS. The dependence of the effect amplitude on the cyclic nucleotides concentrations was obtained. It is suggested that the conductance changes observed are due to the action of cyclic nucleotide dependent protein kinase on the rod outer segment plasma membrane. PMID- 6297619 TI - [Paramagnetic centers of the electron transport chain of higher plant photosynthesis]. PMID- 6297621 TI - [Properties of polyphosphate phosphohydrolase of the "leaky" mutant Neurospora crassa with respect to the specific enzyme]. AB - Some properties of polyphosphate phosphohydrolase from N. crassa strain ad 6.28610a and from its mutant with a decreased polyphosphate phosphohydrolase activity were compared. It was shown that the pH optimum for both enzyme species lies within the range of 7.1-7.3; the temperature optimum is 45 degrees. The mutant polyphosphate phosphohydrolase has a V value, which is 2 times less than that of the parent strain, and possesses a higher thermal inactivation stability. The enzymes of both cultures have practically identical values of Km(app), which depend on the length of the substrate chain. Upon transition from polyphosphate n = 9 to polyphosphate n = 180 the enzyme affinity is increased. Electrophoretic separation of cell-free extract proteins in polyacrylamide gel revealed the existence of two enzyme isoforms in both strains. The electrophoretic mobility of these isoforms are identical in both cultures. Biosynthesis of polyphosphate phosphohydrolase and tripolyphosphatase of N. crassa, unlike that of bacteria, is not controlled by a system common for alkaline phosphatase. PMID- 6297620 TI - [Kinetics of light-induced redox changes of high-potential cytochrome in the chromatophores of purple sulfur bacteria Ectothiorhodospira shaposhnikovii]. AB - Light-induced redox changes of high-potential cytochrome Ch (Em 7.0 = + 290 mV) have been studied. It was found that after switching off the actinic light there is a delay in the cytochrome dark reduction. The extent of the delay depends on the intensity of actinic light, being the more the higher the intensity. A simple kinetic model is proposed to explain both kinetics of redox changes of the cytochrome and dependence of the delay upon the intensity of actinic light. PMID- 6297622 TI - [Interaction of ubisemiquinone with succinate dehydrogenase and the cytochrome chain of mitochondria]. AB - The free radical EPR signals of ubisemiquinone in mitochondria and submitochondrial particles (SMP) were investigated. One of the signals observed under the conditions of the respiratory chain highly oxidized and characterized by an unusually short time of the spin-lattice relaxation has previously been termed as SQ-2. The intensity of SQ-2 in SMP strongly depends on pH, the maximal concentration of QH. is reached at about 8.5. The signal is absent in the succinate dehydrogenase-depleted SMP and is highly sensitive to specific inhibitors of succinate: CoQ-oxidoreductase, such as alpha thenoyltrifluoroacetone and carboxin. In SMP SQ-2 disappears in the presence of low concentrations of ferricyanide, while in mitochondria this non-penetrating oxidant provokes the appearance of SQ-2. The data obtained suggest that SQ-2 belongs to a stable ubisemiquinone which forms a complex with a FeS center of succinate dehydrogenase, is localized at the M-side of the membrane, and is kinetically isolated from the cytochrome chain. Oxidation of the terminal segment of the respiratory chain of mitochondria and SMP reduced by succinate in the presence of antimycin, is in some cases accompanied by an appearance of a strong free radical EPR signal which is stable at 77K but disappears rapidly in the frozen samples at -30- -40 degrees C. It is suggested that the signal is generated by an antimycin-insensitive oxidation of QH2 to QH. via the branch of the respiratory chain comprised of the Rieske FeS-protein and cytochrome c1. The mechanisms of how the two-electron oxidation-reduction of CoQ is coupled with the one-electron transfer through the cytochromes and FeS centers in the respiratory chain are discussed. PMID- 6297624 TI - [Transport of calcium to synaptosomes and subcellular membrane fractions of the brain: effects of opioid peptides]. AB - Highly purified synaptosomal and subcellular fractions identified as mitochondria and microsomes were obtained by fractionation of brain tissues. The greatest Ca accumulating capacity and the highest rate of Ca2+ accumulation were revealed in the mitochondrial fraction. Upon further fractionation of the synaptosomal fraction the energy-dependent uptake (accumulation) of Ca2+ was revealed only in the mitochondria. It was demonstrated that opioid peptides accelerate Ca2+ uptake by the synaptosomes in a medium with physiological concentration of K+ and inhibit this process during K+-dependent membrane depolarization. It was shown that beta-endorphine, methionine-encephaline and leucine-encephaline (10(-8)-10( 5) M) inhibit the Ca-accumulating capacity of both mitochondria and microsomes from brain. The experimental data suggest that opioid peptides can modulate the release of neurotransmitters and/or neurohormones by inhibiting the potential dependent Ca2+ influx into the nerve endings and by decreasing the intrasynaptosomal pool of Ca2+. PMID- 6297623 TI - [cAMP-dependence of phosphorylation of the phosphorylase b kinase of the skeletal muscles of rats during physical exercise and training]. AB - During relative rest of trained rats the phosphorylase b kinase activity is increased by 24% (pH 6.8). Physical load causes an increase of the phosphorylase b kinase activity of untrained and trained rats by 44 and 33%, respectively. The degree of phosphorylase b kinase phosphorylation by a homologous soluble cAMP dependent protein kinase from the muscles of trained rats at rest is 1.9 times that of the control group. The cAMP-dependent phosphorylation of phosphorylase b kinase of untrained and trained rats under physical load is increased 2.5-fold. The data obtained are indicative of the regulatory role of cAMP-dependent phosphorylation in biochemical adaptation of skeletal muscles when their function is increased. PMID- 6297625 TI - [Interaction of spin-labelled phosphatidyl choline with Ca-dependent ATPase from sarcoplasmic reticulum]. AB - The interaction between 3-acyl-2-(6-doxylpalmitoyl) phosphatidyl choline used as a hydrophobic spin probe and Ca2+-dependent ATPase from sarcoplasmic reticulum membranes of rabbit and carp white skeletal muscles was studied. The spin label incorporation into ATPase preparations was performed at initial steps of ATPase isolation by incubating reticulum membranes with the spin probe in the presence of cholic acid. A comparison of the molecular mobility of the probe incorporated into ATPase preparations and into liposomes formed from ATPase phospholipids demonstrated that the presence of the protein in the membrane produces the same effect on the probe mobility as does the decrease of temperature by 10-15 degrees C. The molecular mobility of the probe in the ATPase preparation is increased during protein thermal denaturation. The breaks on the Arrhenius plots for the probe molecular mobility are revealed at the same temperatures (25 degrees for rabbit reticulum and 16 degrees for carp reticulum) as those for the ATPase activity. PMID- 6297626 TI - Activation of adrenal function in fetal sheep by the infusion of adrenocorticotropin to the fetus in utero. AB - We determined whether ACTH1-24, infused into fetal lambs at a rate that is known to cause premature labor, elicits changes in the responsiveness of the fetal adrenal glands, and alters the pattern of corticosteroid output. Plasma cortisol (F), corticosterone (B) and progesterone (P4) were measured during 72 h of infusion of saline or ACTH (10 micrograms/h) beginning on Day 127 of pregnancy. Adrenals were then dispersed into isolated cells, and the output of F, B and P4 after exogenous ACTH determined in vitro. Plasma concentrations of F and B were higher in ACTH-treated fetuses. The increment in F (5-to 7-fold) was greater than that in B (2-fold) such that the F:B ratio in plasma of ACTH-treated fetuses on Days 2 and 3 of infusion was 2.5 times higher than in controls. After 72 h of infusion, the adrenal weights in ACTH-treated fetuses (741 +/- 38 mg, +/- SEM; n = 4) were greater than in the control animals (349 +/- 11 mg). There was a significant effect of ACTH pretreatment in vivo on F output by isolated adrenal cells in vitro. Mean increments in F output after addition of ACTH1-24 (5000 pg/ml) in vitro rose from 368 +/- 235 pg/50,000 cells in controls, to 64,639 +/- 19,875 pg/50,000 cells after ACTH in vivo. There was no significant effect of ACTH in vivo on B output in vitro; the ratio of F:B output, either in the absence or presence of ACTH in vitro, was significantly higher in cells from ACTH pretreated fetuses. There was a significant effect of in vivo ACTH on in vitro P4 output. After ACTH treatment in vivo there was an increase in the vitro output ratio of F:P4, but no change in the output ratio of B:P4. We conclude that ACTH treatment of the fetal lamb in vivo results in activation of fetal adrenal function, increased fetal adrenal responsiveness to ACTH, and directed corticosteroid biosynthesis towards cortisol. Our results are consistent with an increase in fetal adrenal 17 alpha-hydroxylase activity after ACTH treatment. PMID- 6297627 TI - Spontaneous lipid peroxidation in rabbit epididymal spermatozoa: its effect on sperm motility. AB - Rabbit spermatozoa released from the cauda epididymidis into Tris phosphate medium containing KCl or NaCl and 0.4 mM EDTA underwent spontaneous lipid peroxidation during aerobic incubation at 37 degrees C. In the medium containing 130 mM K+ and O mM Na+ (KTP), the rate of lipid peroxidation, as measured by malonaldehyde production, proceeded at a linear rate of 0.045 nmol malonaldehyde/h per 10(8) cells for 22 h. The motility of these spermatozoa declined with time in medium KTP, with 40% initial forward motility decreasing to zero in 4 h and initial 60% flagellar beating ceasing after 12 h. The percent inert spermatozoa showing no flagellar motion in KTP increased linearly with production of malonaldehyde; all flagellar activity stopped at 0.5 nmol malonaldehyde/10(8) cells. In the Tris phosphate medium containing 120 mM Na+ and 10 mM K+ (NTP), the percentage of sperm showing forward motility was close to 100% and this declined to 60% after 16 h aerobic incubation. Flagellar beating was not observed. In medium NTP, the rate of lipid peroxidation was 0.0056 nmol malonaldehyde/h per 10(8) cells, eightfold lower than that observed in KTP. The same linear correlation between malonaldehyde production and percent inert sperm was found as for KTP: 0.5 nmol malonaldehyde/10(8) cells also corresponded to cessation of flagellar motion. The dependence of motility maintenance on K+ concentration in Tris phosphate medium containing (Na+ + K+)=130 mM showed maximal maintenance at 10 mM K+, with a decline at 0 mM K+ and steep decline at K+ concentrations greater than 30 mM. This strong dependence of rabbit sperm peroxidation on ionic composition of the medium is suggested to involve perturbation of the equilibrium between O2 .- and its conjugate acid species being the agent of peroxidation. PMID- 6297628 TI - Production of superoxide and activity of superoxide dismutase in rabbit epididymal spermatozoa. AB - Mature rabbit spermatozoa from the cauda epididymidis suspended in potassium Tris phosphate buffer at 24 degrees C produced O2.-, as measured by reduction of acetylated ferricytochrome c, with an intrinsic rate of 0.20 nmol/min per 10(8) cells. This rate increased to 1.80 nmol/min per 10(8) cells in the presence of 10 mM cyanide. These spermatozoa contain 2.8 units per 10(8) cells of superoxide dismutase activity, 95% of which is sensitive, and 5% of which is insensitive, to cyanide inhibition. These activities correspond to the cytosolic Cu-Zn form and the mitochondrial Mn form of the dismutase, respectively. Only the cyanide sensitive form is released from the sperm on hypo-osmotic treatment or sonication. Hypo-osmotically treated rabbit epididymal spermatozoa produced O2.- with an intrinsic rate of 0.24 nmol/min per 10(8) cells, which increased to 0.58 nmol/min per 10(8) cells in the presence of 10 mM cyanide. Both intact and hypo osmotically treated cells react with O2.- in a second order reaction as inferred from the hyperbolic dependence on cell concentration of O2.- production rate in both the absence and presence of cyanide. The second order rate constant for this reaction with intact cells, kS, was calculated to be 22.9 X 10(-8) (cells/ml)-1 min-1 in its absence. For hypo-osmotically treated cells, the values of kS were 10.8 X 10(-8) (cells/ml)-1 min-1 and 8.2 X 10(-8) (cells/ml) -1 min-1, respectively. Since hypo-osmotically treated cells have lost much of their plasma membrane, the lower value of kS for the treated cells implies that this membrane is one site of reaction of O2.- with the cells. The increase in kS in the presence of cyanide, which inhibits superoxide dismutase and so increases O2.- production, suggests that the cells become more reactive with O2.- as its production rate increase, as would be expected for the occurrence of radical chain oxidation. This in turn suggests that superoxide dismutase plays a major role in protecting rabbit sperm against damage from lipid peroxidation. PMID- 6297629 TI - Mechanism of glucocorticoid-induced suppression of testicular androgen biosynthesis in vitro. AB - The mechanism whereby glucocorticoids directly inhibit gonadotropin-stimulated testosterone production was studied by using primary cultures of testicular cells from adult hypophysectomized rats. Testicular cells were maintained in serum-free media with hormone treatments administered on Day 8 and media collected 48 h later for steroid and cAMP measurement. Highly purified human chorionic gonadotropin (hCG) increased testosterone production relative to controls. Concomitant administration of either natural (cortisone greater than deoxycorticosterone = aldosterone) or synthetic (dexamethasone greater than or equal to prednisolone) corticosteroids inhibited hCG-stimulated testosterone production in a dose-dependent manner. Dexamethasone at 10(-7) M decreased testosterone production by approximately 50-60% and this inhibitory effect was reversible upon removal of the glucocorticoid. In the presence or absence of a phosphodiesterase inhibitor, dexamethasone decreased hCG-stimulated cAMP production by approximately 60%. Dexamethasone also decreased testosterone production induced by cholera toxin and (Bu)2 cAMP by 43 and 63%, respectively. The dexamethasone suppression of testosterone production was accompanied by marked decreases in androstenedione (80% decrease) and 17 alpha hydroxyprogesterone (57%) production, with a lesser effect on progesterone production (28% decrease) and no effect on pregnenolone production. Exogenous progesterone and 17 alpha-hydroxyprogesterone augmented hCG-stimulated testosterone production. Dexamethasone reduced the conversion of exogenous progesterone to testosterone by 33% but did not affect the conversion of 17 alpha hydroxyprogesterone to androstenedione and testosterone, suggesting a specific inhibition of 17 alpha-hydroxylase. These results suggest that glucocorticoids directly suppress Leydig cell steroidogenesis by decreasing gonadotropin stimulation of cAMP production and the activity of 17 alpha-hydroxylase. PMID- 6297630 TI - Studies on the binding of a 32K rat epididymal protein to rat epididymal spermatozoa. AB - A glycoprotein of molecular weight 32K has been isolated and purified from the rat caudal epididymal fluid by gel filtration, ion-exchange and affinity chromatography. The highly purified protein was labeled with radioactive iodine and the binding of the 125I-labeled 32K rat epididymal protein (REP) to washed rat caudal epididymal sperm was studied under various conditions. Scatchard plots of the binding data revealed two binding kinetics. One bound with high affinity (KD = 2.6 X 10(-10) ) but low capacity. The other bound with lower affinity (KD = 2.2 X 10(-9)M) but high capacity. The rate of binding of the labeled protein to sperm was dependent on the temperature of the incubation medium. At the scrotal temperature of 33 degrees C, maximal binding was obtained after 40 min. However, at 22 degrees C equilibrium state was reached after 90 min and at 0 degrees C, the equilibrium rate was not reached even after 120 min of incubation. Binding showed dependence on extracellular pH (optimal pH at 4) and ionic strength of the incubation medium. High ionic strength was found to inhibit binding of the 125I labeled 32K REP to rat caudal epididymal sperm. Specific binding was abolished by 100-fold molar excess unlabeled 32K REP or by native rat caudal epididymal fluid proteins, but not by albumin or ovalbumin. This indicates high specificity of binding. This study has provided direct evidence for the interaction of an epididymal protein with epididymal spermatozoa. PMID- 6297631 TI - Responses to cafeteria feeding in mice after the removal of interscapular brown adipose tissue. AB - Feeding a cafeteria diet to mice resulted in an increased energy intake of approximately 30% and this led to increases in the wet weight, total protein content, and total cytochrome oxidase activity of interscapular and dorso cervical brown adipose tissue. Surgical removal of interscapular brown adipose tissue, followed by cafeteria feeding, gave rise to an elevation in dorso cervical brown adipose tissue wet weight, total protein content, and total cytochrome oxidase activity, compared to intact cafeteria-fed mice. Cafeteria feeding with or without the removal of interscapular brown adipose tissue did not lead to significant increases in body weight compared to stock-fed control mice, but both cafeteria-fed groups of mice showed significant elevations in body fat content indicating that the induced hyperphagia led to a relative obesity in the cafeteria-fed groups. The results presented are consistent with an increased thermogenic activity in the brown adipose tissue of cafeteria-fed mice, and the effect of the removal of interscapular brown adipose tissue further indicates the quantitative importance of the tissue in the control of body weight. PMID- 6297632 TI - Strategies for detecting and characterizing restriction fragment length polymorphisms (RFLP's). PMID- 6297633 TI - Prenatal diagnosis by DNA analysis. PMID- 6297634 TI - [Endogenous activators and inhibitors of Na, K-ATPase induced by acetylcholine]. AB - Preincubation of rat brain homogenates with acetylcholine (ACh) in concentrations of 10(-3)-10(-5) M for 60 minutes produces an essential increment (15-30%) in activity of microsomal Na, K-ATPase. Analogous effect was exerted by the acetylcholinesterase inhibitor eserine (10(-5)-10(-6) M). Acetylcholine has no effect in the presence of actinomycin D. Dialysis of microsomes isolated from the homogenate incubated with ACh leads to a decrease in the enzyme activity and release to the dialysate of low-molecular factor activating Na, K-ATPase of intact microsomes. The latter fact evidences the ACh-induced synthesis of activating factor and inhibition of Na, K-ATPase synthesis. After the animals are administered eserine (0.2-0.4 mg/kg), isolated microsomes show a reduced level of Na, K-ATPase (by 10-15%). Dialysis of microsomes leads to an appreciable elevation (by approximately 40%) of the enzyme activity and release into the dialysate of the inhibitory factor. The differences in the effects of eserine in vivo and in vitro suggest that during the impairment of brain integrity certain effects are excluded from the processes of the control over Na, K-ATPase activity. One of these may involve the impairment of intercellular interactions, for example, the disappearance of the effect on cholinoceptive cells of internuncial neurons that release inhibitory neurotransmitters (catecholamines). PMID- 6297635 TI - Chronic granulomatous disease due to granulocytes with abnormal NADPH oxidase activity and deficient cytochrome-b. AB - A patient with an X-linked genetic disease resembling chronic granulomatous disease (CGD) but differing in several aspects from previously studied cases is described. The oxidase enzyme of the patient's granulocytes was normally activated, but had reduced activity as shown by an increased Michaelis constant and decreased maximum velocity of NADPH-dependent superoxide production. Cytochrome-b was undetectable in dithionite difference spectra. This CGD-like disease further implicates cytochrome-b as an important component of the microbicidal NADPH oxidase system and provides insight into its role in the enzyme complex. PMID- 6297636 TI - Plasma and urine cyclic nucleotide levels in patients with acute and chronic leukemia. AB - Plasma and urine levels of cyclic adenosine 3',5'-monophosphate (cAMP) and of cyclic guanosine 3',5'-monophosphate (cGMP) were measured in 35 normal subjects, in 24 patients with nonneoplastic diseases (iron deficiency anemia, peptic ulcer, and cholelithiasis), and in 50 leukemic patients. The leukemic group included patients with acute lymphoblastic leukemia, acute myelogenous leukemia, chronic lymphocytic leukemia, and chronic myelogenous leukemia. All patients were recently diagnosed and untreated, except for 5 patients with blastic transformation of chronic myelogenous leukemia who had been previously treated. There were no significant differences in plasma and urine cyclic nucleotide levels between normal subjects and patients with nonneoplastic diseases. In leukemic patients, plasma and urine cAMP levels were similar to those of normal subjects, whereas plasma and urine cGMP levels were markedly elevated. There were no significant differences in cGMP values between the various types of leukemia. After starting treatment, plasma cyclic nucleotide levels were periodically measured in 21 of the patients with acute leukemia; cGMP levels were normalized in all the 16 subjects who attained complete remission, whereas both cAMP and cGMP levels were apparently unaffected in the patients who did not respond to treatment. This suggests that plasma or urine cGMP could be used as an additional parameter to monitor the patient's response to treatment. PMID- 6297637 TI - Role of myeloperoxidase in the respiratory burst of human neutrophils. AB - Myeloperoxidase (MPO), a heme enzyme present in the primary granules of polymorphonuclear leukocytes (PMNs), has been demonstrated to participate in the oxygen-dependent microbicidal activity of these cells. Evidence for the importance of MPO in this role comes in part from studies of normal PMNs treated with the heme enzyme inhibitor, sodium azide. MPO has also been suggested to regulate the respiratory activity of PMNs during phagocytosis. The role of MPO in PMN oxygen metabolism was examined by studying parameters of the respiratory burst of PMNs from a number of unrelated MPO-deficient subjects; in addition, the ability of heme enzyme inhibitors to duplicate the MPO-deficient state was studied by treating normal and MPO-deficient cells with these compounds. MPO deficient PMNs were found to have a time-dependent hypermetabolic response as assessed by measurement of oxygen consumption, superoxide generation, hydrogen peroxide release, and hexose monophosphate shunt activity. Catabolic pathways for hydrogen peroxide were normal, suggesting the increased recovery of oxygen metabolites reflects increased production rather than decreased catabolism of H2O2. These observations support the concept that MPO may play an important role in terminating the respiratory burst of normal PMNs. The three heme enzyme inhibitors studied--sodium azide, potassium cyanide, and 3-aminotriazole- differed greatly in the degree to which they inhibited various enzymatic systems in the PMN. Nonetheless, as a group, they exerted qualitatively similar effects on oxygen metabolism of normal and of MPO-deficient PMNs. This indicates that many of the mechanisms by which heme enzyme inhibitors influence PMN metabolism are independent of the inhibition of MPO. Conclusions from studies using such treatment of PMNs should be interpreted with caution. PMID- 6297638 TI - Increase of Na-K-ATPase activity, glutamate, and aspartate uptake in dog erythrocytes associated with hereditary high accumulation of GSH, glutamate, glutamine, and aspartate. AB - We have found convincing evidence for the presence of Na-K-ATPase and high potassium (K) and low sodium (Na) concentrations in the erythrocytes of some dogs associated with hereditary high concentrations of erythrocyte glutathione and some amino acids, glutamate, glutamine, and aspartate. The Na-K-ATPase activity of the erythrocyte membranes of the dogs was about 3 times that of human erythrocyte membranes, whereas the enzyme activity was not detected in control dogs with a normal level of blood glutathione. The Michaelis constant of the enzyme for ATP (Km ATP) was 6.6 X 10(-3)M in the dogs' erythrocytes and 5.0 X 10( 4)M in the human erythrocytes. The concentration of K in the erythrocytes in the dogs examined was about 11 times that of the controls, whereas the erythrocyte Na concentration in the dogs was about one-third that of the controls. The concentrations of K and Na in the plasma of the dogs were equal to those of the controls. Furthermore, L-3H-glutamate and L-3H-aspartate uptake by those cells with high activity of Na-K-ATPase greatly increased, while L-3H-glutamine uptake was unchanged. It appeared that Na+ and K+ gradients created by Na-K-ATPase across the cell membrane might stimulate glutamate and aspartate uptake by the cells, thus causing the high accumulation of such amino acids in the cells. PMID- 6297639 TI - Myeloproliferative syndrome induced by MPSV in DBA/2 mice: presence of a mixed colonies promoting activity (MPA) in the spleen. AB - The myeloproliferative syndrome induced by the myeloproliferative sarcoma virus (MPSV) in DBA/2 mice stimulates the proliferation of pluripotent hemopoietic stem cells (HSC) and of progenitors committed toward granulomacrophagic and erythroid cell lines. This stimulation may result from a direct effect of the MPSV on HSC or from an indirect effect via locally secreted factors. Normal isogenic bone marrow cells were incubated in the mixed colony-forming unit system in semisolid medium supplemented with conditioned media obtained after incubating neoplastic spleen cells for 3 days at 37 degrees C. These spleen conditioned media contain an activity that is physically separable from MPSV by ultracentrifugation and which, in the presence of a very low quantity of erythropoietin, can induce in vitro the proliferation and differentiation of pluripotent HSC, detected by this Mix-CFU technique. We termed this activity mixed-colonies promoting activity (MPA). These results suggest that the hyperplasia of the nonlymphoid hematopoietic system in the neoplastic spleen results from an indirect effect of the MPSV on pluripotent HSC via locally secreted factors. PMID- 6297640 TI - Determination of polynuclear aromatic hydrocarbons in the New York Bight area. PMID- 6297641 TI - Determination of pentachlorophenol volatilized from wood via collection on silica gel. PMID- 6297642 TI - Nonlinear generalizations of the Kedem-Katchalsky equations for ionic fluxes. PMID- 6297643 TI - Genital herpes. PMID- 6297644 TI - Storage of biogenic amines in guinea-pig brain synaptosomes: influence of proton gradient and membrane potential. AB - 1 The effects of K(+), NaCN and the ionophores monensin, nonactin and carbonyl cyanide-p-trifluoro-methoxyphenylhydrazone (FCCP) on the contents of [(3)H]-5 hydroxytryptamine ([(3)H]-5-HT), [(3)H]-dopamine and [(3)H]-noradrenaline ([(3)H] NA) in guinea-pig synaptosomes preloaded with these amines were measured.2 In the presence of Ca(2+), K(+) markedly reduced the amine content of the synaptosomes, indicating an acceleration of spontaneous amine release. In the absence of Ca(2+), K(+) had much less effect.3 Monensin, nonactin and FCCP caused a release of all the three labelled amines. This release was considerably faster and more marked than that induced by K(+) and showed no dependence on Ca(2+). The ionophores did not release lactate-dehydrogenase from synaptosomes.4 NaCN, a blocker of oxidative energy production, did not enhance the spontaneous release of [(3)H]-5-HT nor did it influence the monensin-induced release of [(3)H]-5-HT.5 It is concluded that (a) the intragranular storage of 5-HT, dopamine and NA is dependent on the maintenance of a pH-gradient across the granular membrane as well as on the granular membrane potential; (b) the ionophores cause a non exocytotic release of granular amines, and (c) blood platelets are partial models for aminergic brain neurones as far as intragranular amine storage is concerned. PMID- 6297645 TI - Seasonal variations in drug response and staircase phenomena in atrial muscle from a hibernating rodent (Spermophilus richardsonii). AB - 1 Electrically driven atrial muscle from a hibernating rodent (Spermophilus richardsonii) showed marked seasonal variations in inotropic responsiveness to both the cardiac glycoside actodigin and to raised extracellular calcium concentration. In contrast, glycoside toxicity was apparent and comparable in all tissues. 2 These variations were accompanied by changes in force-frequency ('staircase') characteristics of the tissues. 3 In contrast, no seasonal variation was observed in either positive inotropic responses to rubidium (a non glycoside inhibitor of Na+ + K+ ATP-ase) or negative inotropic responses to methacholine. 4 In summer atria the calcium antagonist, verapamil (6 x 10(-7) M) abolished the positive staircase phenomenon but did not modify the positive inotropic actions of actodigin. 5 These data do not support the hypothesis that 'staircase' and glycoside-induced inotropism are mediated through a common mechanism. PMID- 6297646 TI - Characteristics of GABAB receptor binding sites on rat whole brain synaptic membranes. AB - 1 Saturable binding of (+/-)-[3H]-baclofen and [3H]-gamma- aminobutyric acid ([3H]-GABA) to rat brain crude synaptic membranes has been examined by means of a centrifugation assay. 2 The binding of [3H]-baclofen could be detected in fresh or previously frozen tissue and was dependent on the presence of physiological concentrations of Ca2+ or Mg2+ although a lower affinity Na+ -dependent component could also be observed. Both components probably reflect binding to receptor recognition sites. 3 The saturable portion of bound [3H]-baclofen formed 20.3 +/- 6.9% of total bound ligand. This could be displaced by GABA (IC50 = 0.04 microM), (-)-baclofen (0.04 microM) and to a much lesser extent by (+)-baclofen (33 microM). Isoguvacine, piperidine-4-sulphonic acid and bicuculline methobromide were inactive (up to 100 microM) and muscimol was only weakly active (IC50 = 12.3 microM). 4 Saturable binding of [3H]-GABA increased on adding CaCl2 or MgSO4 (up to 2.5 mM and 5.0 mM respectively) to the Tris-HCl incubation solution. This binding (GABAB site binding) was additional to the bicuculline-sensitive binding of GABA (GABAA site binding) and could be completely displaced by (-)-baclofen (IC50 = 0.13 microM). 5 Increasing the Ca2+ concentration (0 to 2.5 mM) increased the binding capacity of the membranes without changing their affinity for the ligand. 6 The binding of [3H]-GABA to GABAB sites could be demonstrated in fresh as well as previously frozen membranes with a doubling of the affinity being produced by freezing. Further incubation with the non-ionic detergent Triton-X 100 (0.05% v/v) reduced the binding capacity by 50%. 7 The pharmacological profile of displacers of [3H]-GABA from GABAB sites correlated well with that for [3H]-baclofen displacement. A correlation with data previously obtained in isolated preparations of rat atria and mouse vas deferens was also apparent. 8 It is concluded that [3H]-baclofen or [3H]-GABA are both ligands for the same bicuculline-insensitive, divalent cation-dependent binding sites in the rat brain. PMID- 6297647 TI - Evidence for an A1-adenosine receptor in the guinea-pig atrium. AB - 1 The purpose of this study was to determine whether the adenosine receptor that mediates a decrease in the force of contraction of the guinea-pig atrium is of the A1- or A2-sub-type. 2 Concentration-response curves to adenosine and a number of 5'- and N6-substituted analogues were constructed and the order of potency of the purines was: 5'-N-cyclopropylcarboxamide adenosine (NCPCA) = 5'-N ethylcarboxamide adenosine (NECA) greater than N6cyclohexyladenosine (CHA) greater than L-N6-phenylisopropyl adenosine (L-PIA) = 2-chloroadenosine- greater than adenosine greater than D-N6-phenylisopropyl adenosine (D-PIA). 3 The difference in potency between the stereoisomers D- and L-PIA was over 100 fold. 4 The adenosine transport inhibitor, dipyridamole, potentiated submaximal responses to adenosine but had no significant effect on those evoked by the other purines. 5 Theophylline antagonized responses evoked by all purines, and with D-PIA revealed a positive inotropic effect that was abolished by atenolol. 6 The results indicate the existence of an adenosine A1-receptor in the guinea-pig atrium. PMID- 6297648 TI - Stimulation-evoked release of [3H]-noradrenaline by 1, 10 or 100 pulses and its modification through presynaptic alpha 2-adrenoceptors. AB - 1 Mice isolated vasa deferentia were loaded with 1-[7,8-3H]-noradrenaline and subsequently field stimulated with 1, 10 or 100 pulses (2 ms pulse width, 1 Hz). The tritium overflow was separated into [3H]-noradrenaline and its 3H metabolites. 2 The resting release of tritium contained about 7% [3H] noradrenaline, 33% [3H]-3, 4-dihydroxyphenylglycol ([3H]-DOPEG) and 60% 3H-non catechols with usually less than 1% [3H]-dihydroxymandelic acid ([3H]-DOMA). The proportion of the tritium as [3H]-noradrenaline increased with stimulation train length to 35% with 100 pulses; this increase in [3H]-noradrenaline was associated with falls in [3H]-DOPEG and 3H-non-catechols. Generally the proportional increase in [3H]-noradrenaline on stimulation was about 10 x total tritium when compared with the resting release. 3 The fractional release of [3H]-noradrenaline per pulse was independent of train length, averaging about 6 x 10(-6). This was reduced by the alpha 2-adrenoceptor agonist clonidine (0.3 - 30 nM) with an IC50 of 4.8 nM (10 pulses at 1 Hz). 4 The alpha 2-adrenoceptor antagonist, yohimbine (10 - 100 nM), did not alter the fractional release of [3H]-noradrenaline elicited by 1 pulse. The antagonist did not change the amount or composition of the resting or evoked tritium overflow. However, yohimbine (1 - 100 nM) increased the fractional release of [3H]-noradrenaline per pulse for trains of 10 or 100 pulses (1 Hz) in a concentration-dependent fashion. An increase above controls was significant only with 100 pulses and yohimbine, 30 nM. 5 The results show that the release of noradrenaline during trains of pulses in the mouse vas deferens can be regulated through presynaptic alpha 2-adrenoceptors. There was no evidence of inhibition by noradrenaline of its own release following a single pulse. PMID- 6297649 TI - Neostigmine augments responses of the rat anococcygeus muscle to field stimulation. AB - 1 The effects of neostigmine on noradrenergic transmission have been studied in the field stimulated, isolated anococcygeus muscle of the rat. 2 In those muscles where the excitatory response to field stimulation was not completely inhibited by guanethidine (5 X 10(-6) to 10(-5) M) or phentolamine (10(-6) M), atropine (5 X 10(-8) M) gave no further inhibition of the response. 3 The shape of the response to field stimulation was altered in a dose-dependent manner by neostigmine (5 X 10(-7) to 5 X 10(-6) M), such that a 'shoulder' appeared during the relaxation phase. The 'shoulder', present at all stimulation frequencies tested between 3 and 40 Hz, was abolished by atropine (5 X 10(-8) M) and unaffected by tubocurarine (10(-6) M). 4 Neostigmine (2.5 X 10(-6) M), whether alone or in the presence of atropine (5 X 10(-8) M), had no effect on the uptake or stimulation-induced release of [3H]-noradrenaline. 5 Using electron microscopy, small Schwann/axon bundles close to smooth muscle cells rarely showed cholinesterase staining, whereas larger bundles at the outer serosal aspects of the muscle exhibited dense staining. 6 It is concluded that the observed effects of neostigmine are not due to a presynaptic effect on noradrenergic transmission. PMID- 6297652 TI - Sclerotic bone deposits in multiple myeloma. PMID- 6297650 TI - Antagonism by antidepressants of muscarinic acetylcholine receptors of human brain. AB - 1 Twenty-two compounds classified as antidepressants, metabolites of antidepressants or putative antidepressants were assayed for their ability to antagonize the binding of (-)-[3H]-quinuclidinyl benzilate to muscarinic receptors in homogenates of human caudate nucleus. 2 Sixteen of these compounds were assayed for their ability to antagonize carbachol-stimulated cyclic guanosine 3',5'-monophosphate (cyclic GMP) synthesis by intact murine neuroblastoma cells (clone N1E-115). 3 Equilibrium dissociation constants (KDs) for these drugs and the muscarinic receptors of human brain spanned over 4 orders of magnitude, with the tertiary amine tricyclic antidepressant, amitriptyline (KD = 18 nM) being the most potent compound tested and trazodone (KD = 324 microM) the least potent. 4 There was a significant correlation between the data for human and murine receptors and for eight compounds (imipramine, desipramine, maprotiline, mianserin, 3-chloro-2-hydroxyimipramine, amoxapine, 2 hydroxyimipramine and iprindole). KD values measured by the two techniques were not significantly different. PMID- 6297651 TI - The limitations of the dual radionuclide subtraction technique for the external detection of tumours by radioiodine-labelled antibodies. AB - A dual radionuclide subtraction technique for external detection of tumours has been evaluated to determine the viability of the method for use with radioisotope labelled antibodies. A number of external scintigraphic investigations have been carried out with 131I-labelled antibodies to carcinoembryonic antigen (CEA). The investigations were performed on patients with metastatic disease known to produce CEA. The dual radionuclide subtraction technique was used to account for the blood and tissue background. The 131I-labelled antibodies were found to localise in the metastatic lesions, but the subtraction technique using 99Tcm labelled HSA and pertechnetate gave ambiguous results, which included the production of artefacts. The ambiguities noted in the clinical results were substantiated by experimental data, which highlight the unreliability of this technique. PMID- 6297653 TI - Damage profile in rat thyroids after irradiation with a proton microbeam in vivo. PMID- 6297654 TI - Bilateral germinal cell tumour in a 30-year-old man complaining of subfertility. PMID- 6297655 TI - Ketoconazole. PMID- 6297656 TI - Mesotheliomas and asbestos type in asbestos textile workers: a study of lung contents. AB - The asbestos contents of the lungs of former employees of an asbestos textile factory were determined at necropsy using a transmission electron microscope. Those who had died of mesothelioma were compared with a matched sample of those who had died of other causes. The predominant fibre processed in the factory was chrysotile, but crocidolite had also been used. The lung content was consistent with the known exposure to chrysotile, but the crocidolite content was also high, being about 300 times that of the general population of the United Kingdom. The lungs of those with mesothelioma did not contain more of either chrysotile or crocidolite than the lungs of the controls, so no particular type of asbestos could be implicated in causing the mesotheliomas. The evidence of substantial exposure to crocidolite means that the mesotheliomas that occurred in this factory could not be attributed with any certainty to the exposure to chrysotile. PMID- 6297657 TI - Segmental necrotising glomerulonephritis with antineutrophil antibody: possible arbovirus aetiology? PMID- 6297658 TI - Simultaneous primary infections with Epstein-Barr virus and measles virus in fatal acute encephalitis. PMID- 6297659 TI - Coxsackie B infection and arthritis. PMID- 6297660 TI - Repeated renal failure with captopril. PMID- 6297661 TI - Cannabinoids as antiemetics. PMID- 6297662 TI - Characteristics of nasal and temporal retina in Siamese and normally pigmented cats: ganglion cell composition, axon trajectory and laterality of projection. PMID- 6297663 TI - The accessory optic system in the newt, Triturus cristatus: unitary response properties from the basal optic neuropil. AB - Recordings from the basal optic neuropil (BON) demonstrated directional sensitivity. These directional sensitivities were different in different units, however, most of them displayed a strong vertical component. Inhibition in the non-preferred direction did not occur, only a lower activation than in the preferred direction was found. The identified velocity functions correlate well with the velocity dependence of the nystagmus frequency demonstrated in the behavior. The units responded well to moving large-area stimuli and exhibited horizontally elongated receptive fields with diameters up to 120 degrees. Additionally, bimodal recordings were observed in the BON which were probably due to input from the otoliths. As the recordings are likely to be presynaptic, the BON receives types of input which establish its function in the control of visuomotor behavior. PMID- 6297664 TI - Specific binding of [3H]phencyclidine in rat central nervous tissue: further characterization and technical considerations. AB - The interaction of phencyclidine (PCP) with its specific receptor sites in the central nervous system has been further characterized. Kinetic association and dissociation rate constants of 2.9 X 10(6) M-1 and 4.8 X 10(-1) min-1 were determined, yielding a kinetic KD of 1.6 X 10(-7) M, in agreement with the KD previously determined at equilibrium. Permissible separation time of 13 s was calculated from the kinetic data, well above the actual separation time of less than 10 s in the rapid filtration assay. Presoaking of filters in 0.01% poly-L lysine eliminated displacable [3H]PCP adsorption to filter material. Binding data obtained via centrifugation assays was identical to that obtained with the rapid filtration method. Stereospecificity of the PCP receptor was demonstrated by the finding that (+)-ketamine is four-fold more potent than (-)-ketamine in displacing specifically bound [3H]PCP. Several proteolytic enzymes including trypsin, papain and thermolysin potently inactivated PCP receptors. Detailed regional distribution studies showed highest density of PCP receptors in subicular cortex and hippocampus, intermediate levels in hypothalamus, striatum, frontal cortex and cerebellum, lower levels in brainstem and spinal cord, and negligible levels in corpus callosum, a white-matter control area. Benzomorphan opiates with PCP-like behavioral effects interact with the PCP receptor. These data support the pharmacological relevance of the PCP receptor site as demonstrated by the rapid filtration method. PMID- 6297665 TI - Effect of chronic ethanol consumption on central and peripheral type benzodiazepine binding sites in the mouse brain. AB - Following chronic exposure of C57/BL6 mice to ethanol the binding of [3H]Ro5-4864 and [3H]propyl-beta-carboline-3-carboxylate to benzodiazepine binding sites in the brain was studied. Peripheral-type benzodiazepine binding sites were measured using the probe [3H]Ro5-4864. Chronic ethanol treatment resulted in a significant increase in [3H]Ro5-4864 binding due to a 43% increase in receptor density. The affinity of [3H]Ro5-4864 for the receptor was not significantly affected. The binding of [3H]propyl-beta-carboline-3-carboxylate to central-type benzodiazepine receptors was not affected by chronic ethanol treatment. PMID- 6297666 TI - Brainstem relay of disynaptic pyramidal EPSPs to neck motoneurons in the cat. AB - A short train of stimuli delivered to the contralateral pyramid evoked EPSPs with disynaptic latencies in neck motoneurons. After a transection of the pyramid at the obex, stimulation rostral to the lesion still evoked disynaptic EPSPs. Stimulation of the pyramid rostral to a transection at the level of the trapezoid body did not produce synaptic effects. It is suggested that the disynaptic EPSPs in neck motoneurons from the contralateral pyramid are mediated by medullary reticulospinal neurons. PMID- 6297667 TI - Convergence on reticulospinal neurons mediating contralateral pyramidal disynaptic EPSPs to neck motoneurons. AB - Convergence upon reticulospinal neurons which mediate disynaptic, contralateral pyramidal EPSPs to neck motoneurons has been examined in cats with contralateral pyramidal transection at the obex. Conditioning stimuli in the contralateral tectum and ipsilateral mesencephalic tegmentum produced monosynaptic facilitation of the disynaptic pyramidal EPSP, whereas facilitation evoked from the ipsilateral pyramid showed a disynaptic time course. These results show that contralateral pyramidal, tectal and ipsilateral tegmental fibers converge onto common reticulospinal neurons which have direct excitatory connections with neck motoneurons. PMID- 6297668 TI - Alpha-methylDOPA dissociates hypertension, cardiovascular reactivity and emotional behavior in spontaneously hypertensive rats. PMID- 6297669 TI - Neuronal organization of the vestibulospinal system in the cat. AB - In the lateral and descending vestibular nucleus, vestibulospinal neurons were surveyed extra- and intracellularly in cats anesthetized with sodium pentobarbital. The neurons were investigated both by their antidromic activation from the spinal cord (C1, C4, T1, L1 and L5 spinal levels) and from the oculomotor nucleus region, and by orthodromic activation from the vestibular nerve. Axonal courses of vestibulospinal neurons were determined electrophysiologically at C1 level, as medial (MVST) or lateral (LVST). By single and the same electrodes a number of neurons were recorded in wide regions of the lateral and descending vestibular nucleus from single cats. Thus, it became possible to investigate somatotopical localization systematically for the first time with microelectrode techniques. Neurons of origin of the LVST were localized in the lateral vestibular nucleus. Second-order vestibular neurons were localized in the ventral region of the lateral vestibular nucleus, and in the rostral region of the descending vestibular nucleus. Many second-order, double discharge MVST neurons were identified in the descending and lateral vestibular nucleus. Somatotopical localization were recognized, but non-second-order cervical neurons were identified in the dorsal region, although second-order lumbar neurons were identified in the ventral region of the lateral vestibular nucleus. Specific modes of vestibular activation of these vestibulospinal neurons were discussed, and the vestibulospinal systems in the cat and rabbit were discussed. PMID- 6297670 TI - Reformulation of the Papez circuit: absence of hippocampal influence on cingulate cortex unit activity in the primate. AB - Cingulate cortex (CC) unit responses to hippocampal volleys and afterdischarges were studied in 4 awake unsedated squirrel monkeys using extracellular microelectrodes. Stimuli were delivered via bipolar electrodes chronically implanted bilaterally in anterior and posterior hippocampus. A total of 412 units in 21 tracks were studied and tested to single and multiple shock stimulation. Spontaneous firing rates were approximately equally distributed among slow (0.25 spikes/s), medium (2.5-10), and fast (10-25). Only 2% of cingulate units (9/412) responded orthodromically to hippocampal stimulation: 5 with initial excitation; 4 with initial inhibition. Eight responses had initial latencies from 100-200 ms; one unit a latency of 25 ms. Three units displayed periodic, almost cyclical, excitation. All unit response thresholds were 1 shock, 0.25 mA or less. Hippocampal afterdischarge activity influenced the firing pattern of 17 units: 16 showed intraictal excitation and 1 inhibition. In comparison with the preceding studies in the same model these results indicate that the influence of hippocampal stimulation on cingulate cortex is not nearly as prominent as either the fornix or non-fornix hippocampal influence on hypothalamic, preoptic, and basal forebrain structures. It is proposed that the cingulate cortex is not a prominent link in the Papez circuit. PMID- 6297671 TI - Recovery of function and basal ganglia [14C]2-deoxyglucose uptake after nigrostriatal injury. AB - Rats with unilateral 6-hydroxydopamine injections along the mesotelencephalic dopaminergic projection showed a profound impairment in localizing somatosensory stimuli on the contralateral body surface at 3 days postoperatively. Approximately one-half of the affected animals recovered the ability to localize tactile stimuli during 6 weeks postoperatively, whereas the remainder did not. When it occurred, the recovery of sensorimotor function began between the third and fifth day postoperatively and plateaued between days 14 and 21. Unilateral damage to these dopaminergic neurons resulted in hemispheric asymmetries of [14C]2-deoxyglucose incorporation at 3 days postoperatively. For structures that normally receive a dopaminergic innervation (e.g., neostriatum, nucleus accumbens septi, olfactory tubercle) the autoradiographic density of the injured side was decreased relative to the intact hemisphere. For structures that receive striatal inputs (globus pallidus, entopeduncular nucleus, substantia nigra pars reticulata), the autoradiographic density was increased on the side of the injury. This pattern of altered [14C]2-deoxyglucose incorporation was still present at 6 weeks postoperatively in animals that showed no recovery of somatosensory localization during that time. In contrast, rats that did recover showed no hemispheric asymmetries within the anterior neostriatum, globus pallidus, or substantia nigra pars reticulata at 6 weeks postoperatively, and the time course of normalization of metabolic activity in these structures was similar to that for behavioral restoration. These results directly demonstrate the importance of the neostriatum and particular structures efferent to it in the recovery of sensorimotor functions after striatal dopamine depletion. The types of neuronal plasticity within this basal ganglia circuitry responsible for the normalization of [14C]2-deoxyglucose incorporation and behavior are discussed. PMID- 6297672 TI - Recovery of function after mesotelencephalic dopaminergic injury in senescence. AB - After intracerebral 6-hydroxydopamine injections that extensively damage the ascending mesotelencephalic dopaminergic projection, young adult rats develop severe sensorimotor deficits, including an inability to orient toward somatosensory stimuli. Many rats with such damage recover their localization of touch, and this recovery depends on the survival of a small proportion of the neostriatal dopaminergic terminals. Behavioral recovery appears to be mediated by an increased dopamine synthesis and release within surviving terminals and an increased responsiveness to dopamine of neostriatal neurons. The ability of aged rodents to increase activity at neostriatal dopaminergic synapses following partial injury to these neurons is remarkably intact, despite the reduced basal level of transmission at these synapses in senescence. First, 27-28-month-old rats recovered somatosensory localization after injury as readily as did 4-5 month-old animals that had equivalent neostriatal dopamine depletions. Second, old and young rats developed a similar degree of behavioral supersensitivity to apomorphine (0.25 mg/kg, i.p.) after unilateral 6-hydroxydopamine injections, as measured by contralateral turning. Third, injury to these neurons produced an equivalent increase in [3H]spiroperidol binding in the neostriatum of old and young rats at 5-7 weeks. Old rats suffered more extensive neostriatal dopamine depletions after intracerebral 6-hydroxydopamine injections than did young adult animals, particularly when small quantities (2 micrograms) of the neurotoxin were injected. This enhanced susceptibility to 6-hydroxydopamine of dopaminergic neurons in old rats could not be attributed to age differences in the placement of the injection cannula or the extent of the non-specific damage at the injection site. The implications of this enhanced susceptibility to the neurotoxin are discussed. PMID- 6297673 TI - Presence, distribution and pharmacology of conjugated catecholamines in the rat spinal cord. AB - A method is described for the assay of conjugated catecholamines in the brain and spinal cord. Employing this method, the distribution of conjugated norepinephrine (NE) and dopamine (DA) was evaluated in the 5 main regions of the rat spinal cord and compared with those in the hypothalamus. In the spinal cord 26-35% of total catecholamines are conjugated. The percent of NE conjugated in the hypothalamus NE is about 17%, while that of DA is around 12%. Acute treatments with amphetamine, desmethylimipramine and haloperidol failed to change the concentrations of both total and conjugated NE and DA in the cervical spinal cord. Only amphetamine increased hypothalamic conjugated NE. It is concluded that evaluation of central conjugated catecholamines may provide an additional dimension to our ability to critically assess the mechanisms of action of drugs on central catecholamines. PMID- 6297675 TI - Sleep-related variations of membrane potential in the lateral geniculate body relay neurons of the cat. AB - Membrane potential of lateral geniculate body relay neurons was monitored in chronic cats during the sleep-waking cycle. Neurons were tonically depolarized throughout paradoxical (P) sleep and the maximal level of polarization occurred during slow (S) sleep (mean difference of membrane potential between S and P sleep: + 10.2 +/- 1.3 mV, n = 6, range: 8-12 mV). Some features of the spontaneous activity of S and P sleep are briefly discussed in relation to the level of membrane potential. In particular it is suggested that the phasic depolarizations underlying the bursts of action potentials during S sleep, and which are reproduced retinal cell axons impinging upon the hyperpolarized membrane. PMID- 6297674 TI - Brain angiotensinogen: in vitro synthesis and chromatographic characterization. AB - The capacity of rat brain to synthesize angiotensinogen (renin substrate) was examined in vitro using brain slices (300 micrometers thick) incubated in minimum essential medium (Eagles) at pH 7.4 and 37 degrees C for 2 h. The concentration of angiotensinogen (measured as equivalents of angiotensin I;AI) increased from 5.2 +/- 1.1 ng AI/g of incubated tissue to 40 +/- 4 ng AI/g at 2 h. In the presence of cycloheximide (10 micrograms/ml) or vinblastine sulfate (10 micrograms/ml) the values at the 2 h interval were 24.5 +/- 3.5 ng AI/g and 10.5 +/- 3.5 ng AI/g respectively. The angiotensinogen released by brain slices was pooled and some of its physicochemical properties examined and compared to those of angiotensinogen from plasma and saline-perfused brains. Chromatography on Sephacryl S-200 showed brain and plasma angiotensinogen to be similar in size (mol. wt. approximately 59,000) but large differences in binding activity to Concanavalin A were found. Only 31% of plasma angiotensinogen bound to the affinity medium compared to 63% for the pooled incubation medium. It was concluded that de novo synthesis of angiotensinogen occurs in the rat brain and that the brain protein may differ in its glycoside composition from hepatic angiotensinogen. PMID- 6297676 TI - Dorsal root potentials in the cat: effects of bicuculline. AB - Bicuculline (0.2 1 mg/kg) administered intravenously depressed dorsal root potentials (DRPs) evoked by stimulation of mixed, pure muscle or pure cutaneous nerves which was clearly concurrent with enhanced background potentials in intact cat. Administration of sodium pentobarbitone (15-30 mg/kg i.v.) reduced the ability of bicuculline to enhance background potentials and to depress evoked DRPs. In spinalized preparations, bicuculline depression of evoked DRPs by bicuculline in intact cat may not result from its action at axo-axonic GABAergic synapses alone and occlusion may also play a part. However, the role of gamma aminobutyric acid (GABA) in primary afferent depolarization is confirmed in the spinalized preparations. PMID- 6297677 TI - Taurine-induced increase of the Cl-conductance of cerebellar Purkinje cell dendrites in vitro. AB - Whether taurine increases the Cl-conductance of cerebellar Purkinje cell dendrites was examined by intradendritic recording technique in vitro. Taurine induced hyperpolarization was inverted to depolarization by lowering the concentration of external Cl and also by hyperpolarizing the dendritic membrane by intradendritic DC current injection. The reversal potential for the taurine action was found to be linearly related to the logarithmic concentrations of external Cl, the slope being 59 mV for a 10-fold change of external Cl concentration. These results suggest that taurine increases a Cl-conductance for exerting its inhibitory action on cerebellar Purkinje cell dendrites. This finding may support the transmitter role of taurine in the mammalian cerebellum. PMID- 6297678 TI - Activation of dopamine-containing amacrine cells of retina: light-induced increase of acidic dopamine metabolites. AB - The acidic metabolites of dopamine, homovanillic acid (HVA) and 3,4 dihydroxyphenylacetic acid (DOPAC), are present in rat retina. DOPAC is the most abundant metabolite. Both metabolites increase in parallel when rats are taken from a dark to a lighted environment. Haloperidol treatment also increases the metabolites in both dark and light while apomorphine decreases both metabolites in dark and light and partially antagonizes the increase induced by haloperidol. PMID- 6297679 TI - Facilitatory interaction between cutaneous afferents from low threshold mechanoreceptors and nociceptors in segmental reflex pathways to alpha motoneurons. AB - EPSPs evoked by air puff stimulation of the hairy skin of the hindlimb or by low strength stimulation of the lateral sural nerve were facilitated by radiant heat of 50-53 degrees C but not by radiant heat of 42-43 degrees C. The results indicate that afferents from low threshold mechanoreceptors and from nociceptors of the skin converge onto common interneurons in reflex pathways to alpha motoneurons. This would allow a functionally useful cooperation of these two types of afferents in spinal motor control. PMID- 6297680 TI - Oxygen toxicity of hippocampal tissue in vitro. AB - The effects of hyperbaric oxygen have been studied on synaptic transmission in the hippocampal brain slice. Oxygen enhances both the prevolley and postsynaptic potential of extracellularly recorded responses following focal stimulation of afferent fiber bundles, but synaptic efficiency is reduced. It is suggested that hyperbaric oxygen causes changes in potassium distribution in the slice. PMID- 6297681 TI - Opioid and non-opioid mechanisms of stress analgesia: lack of cross-tolerance between stressors. AB - Qualitatively different analgesic responses can be evoked in rats by exposure to prolonged, intermittent or brief, continuous footshock stress. These two forms of stress analgesia appear to be mediated by opioid and nonopioid pain-inhibitory substrates, respectively. The present study confirms our previous observation that tolerance develops to only the opioid form of stress analgesia and shows that cross-tolerance does not occur between the opioid and nonopioid forms. These data provide further evidence that independent mechanisms underlie opioid and nonopioid stress analgesia. PMID- 6297682 TI - Lesion of a serotonergic modulatory neuron in Aplysia produces a specific defect in feeding behavior. AB - The serotonergic metacerebral cells (MCCs) of Aplysia were destroyed by intracellular injection of proteolytic enzyme. MCC-lesioned animals showed alterations of biting responses compared to MCC-sham and B-cell-lesioned control animals, as well as to their own preoperative behavior. The alterations of biting responses included a prolongation of the duration of radula protraction and a lengthening of interbite interval. No changes were observed in non-biting feeding responses and in behaviors unrelated to feeding. PMID- 6297683 TI - Regulation of the strength of adenosine modulation in the hippocampus by a differential distribution of the density of A1 receptors. AB - Correlative studies examining: (1) the binding of [3H]cyclohexyladenosine to A1 adenosine receptor sites, and (2) the depressive action of adenosine on evoked activity, were performed in the hippocampal formation. A greater number of A1 sites was observed in the dorsal versus ventral aspect of the hippocampus, and a larger depression of evoked potentials by adenosine was obtained in the dorsal aspect. These observations suggest that a differential density of A1 receptors might control the magnitude of modulatory action by adenosine on hippocampal circuitry. PMID- 6297684 TI - Octopamine in Lumbricus terrestris: presence, synthesis and effect of octopamine on the spontaneous rhythmic contractions of the ventral nerve cord. AB - A radiochemical-enzymatic assay was utilized to measure endogenous levels of octopamine in the nerve cord and blood of the earthworm. The results show the presence of octopamine in the ventral nerve cord and blood at concentrations of 2.79 +/- 0.5 (means +/- S.D.) ng/mg wet tissue weight and 9.5 +/- 1.6 x 10(-8) M (means +/- S.D.) respectively. The ability of the ventral nerve cord to synthesize octopamine was investigated by incubating the tissue in vitro in radiolabeled percursors. The results show the synthesis of [3H]octopamine from both precursor [3H]tyrosine and [3H]tyramine. In vitro incubation of the isolated ventral nerve cord in physiological concentrations of octopamine revealed modulation of the spontaneous rhythmic contractions of the nerve cord. The data provide direct support for a modulatory role of octopamine in L. terrestris. PMID- 6297685 TI - Behavior and binding: correlations between alpha 1-adrenergic stimulation of acoustic startle and alpha 1-adrenoceptor occupancy and number in rat lumbar spinal cord. AB - The relationship between alterations in alpha 1-adrenoceptors and behavioral effects of alpha 1-adrenergic agonists were investigated in a localized region of the rat central nervous system. Direct infusion of the alpha 1-adrenergic agonists, D-amphetamine or phenylephrine. into the subarachnoid space of the lumbar cord (intrathecal administration) increased the amplitude of the acoustic startle reflex, The magnitude of this behavioral facilitation correlated highly with the degree of alpha 1-adrenoceptor occupation measured by [3H]prazosin binding in lumbar spinal tissue. Using an in vitro estimate of receptor occupation, maximal potentiation of startle occurred following approximately 30% occupation of the receptors, using either D-amphetamine or phenylephrine. Intrathecal administration of 6-OHDA produced a 95% decrease in spinal norepinephrine and markedly enhanced the behavioral response to intrathecal phenylephrine as well as the number of alpha 1-adrenoceptors. The correlation between the time course of the behavioral and binding changes was 0.99. No change in receptor affinity (KD) or receptor occupation by phenylephrine was found after 6-OHDA. The data indicate that receptor binding parameters do have predictive value for behavior, especially if localized regions of the nervous system, critical to the behavior, are analyzed. PMID- 6297686 TI - Biological and immunological characterization of alpha-melanocyte-stimulating hormone (alpha-MSH) in two neuronal systems of the rat brain. AB - Recent immunocytochemical studies have demonstrated the existence of two different neuronal systems containing alpha-MSH-like material in the brain: one originating from the arcuate nucleus and the other one from the dorsolateral hypothalamus. The aim of the present study was to further characterize alpha-MSH in these two discrete regions of the rat diencephalon. Intracerebroventricular administration of colchicine resulted in a marked decrease in the number of ACTH and beta-endorphin nerve fibers and a significant reduction in ACTH and beta endorphin content in the dorsolateral hypothalamus. Conversely, colchicine treatment did not alter alpha-MSH, ACTH or beta-endorphin content in the arcuate nucleus and did not significantly affect alpha-MSH concentration in the dorsal region. Reverse-phase high-performance liquid chromatography showed that the major alpha-MSH-like compound localized in the dorsal hypothalamus co-migrated exactly with synthetic alpha-MSH, whereas the arcuate nucleus contained 5 peptides cross-reacting with alpha-MSH antibodies, 4 of them being different from standard alpha-MSH. Significant amounts of biologically active melanotropin, which migrated on Sephadex G-25 columns like synthetic alpha-MSH, were also detected in both the arcuate nucleus and dorsolateral hypothalamus. Taken together, these data demonstrate that the alpha-MSH cell bodies located in the dorsolateral hypothalamus specifically produce authentic alpha-MSH, whereas the alpha-MSH cell bodies in the arcuate nucleus also contain ACTH, beta-endorphin and several peptides immunologically related but not identical to alpha-MSH. PMID- 6297687 TI - Reserpine inhibits rat anterior pituitary hormone secretion in vitro: effects on prolactin and ACTH and ultrastructural observations. AB - We measured [3H]prolactin ([3H]Prl) synthesis and secretion in female rat anterior hemipituitary glands incubated in vitro, and immunoassayable Prl secretion from dispersed anterior pituitary cells in a perfused column. Anterior pituitary glands which were incubated in 9 microM reserpine showed a marked inhibition of [3H]Prl secretion but no change in hormone synthesis, thus causing [3H]Prl accumulation within the gland. The same concentration of reserpine produced a similar effect in pituitary glands taken from rats depleted of dopamine with alpha-methyl-p-tyrosine. Reserpine inhibited Prl secretion from dispersed anterior pituitary cells with a gradual onset and prolonged duration. Thyrotropin-releasing hormone (TRH), but not dibutyryl cyclic AMP (dbcAMP), the calcium ionophore A23187 or excess Ca2+, stimulated both [3H]Prl and Prl secretion in the presence of reserpine. In contrast, neither basal nor vasopressin-stimulated ACTH (bio- and immunoassayable) secretion was inhibited by 9 microM reserpine. Ultrastructurally, pituitary glands incubated in reserpine had an increased content of Prl secretory granules. Reserpine thus selectively inhibited Prl secretion, secondarily causing accumulation of both measurable hormone and Prl secretory granules within the pituitary gland. We hypothesize that reserpine interrupted calcium-dependent mechanisms in the stimulus-secretion coupling process to inhibit Prl release. PMID- 6297688 TI - Effect of intestinal amino acid infusions on hypothalamic single unit activity in the anesthetized cat. AB - Single unit discharges in the ventromedial hypothalamic nucleus (VMH) and lateral hypothalamic area (LH) were recorded extracellularly in anesthetized cats, while amino acid solutions were perfused through the small intestine via implanted cannulae. Test infusions consisted of 5 amino acid mixtures (arginine, leucine, phenylalanine, tryptophan, alanine: 50 mM each), 5.0% casein hydrolysate; arginine 125 mM (2.2%); leucine 125 mM (1.6%). Sixty-six units were recorded in 30 cats. Of 52 tested in the LH, 19 or 37% were affected: 14 increased and 5 decreased in firing rate in response to amino acid intestinal perfusions with a very short latency consistent with activation of intestinal amino acid receptors. Of 14 neurons tested in VMH, only 3 or 21% were activated by amino acid infusions but with a long latency of several minutes which cannot exclude the involvement of a postabsorptive signal. Results are discussed in terms of intestinal afferent control over hypothalamic neuronal activity related to amino acid induced satiety. PMID- 6297689 TI - alpha- and beta-adrenergic binding sites in sheep cerebral cortex: characterisation, effects of photoperiod and treatment with estrogen/progesterone. AB - The radioligands [3H]-dihydroergocryptine and [3H]-dihydroalprenolol were used to characterise alpha- and beta-adrenergic binding sites, respectively, in membrane fractions of sheep cerebral cortex. In terms of affinity, density and specificity these sites possess properties similar to those previously characterised in rat brain. Further, in preliminary studies, these sites also appear to be responsive to treatment with estradiol/progesterone as well as to photoperiod. Thus, estrogen treatment can elevate both alpha- and beta-adrenergic binding sites in cortical tissue of sheep kept in natural light. In contrast, artificial light either has no effect or inhibits binding in response to estrogen. PMID- 6297690 TI - Blockade of stress-induced prolactin release in monosodium glutamate-treated rats. AB - To elucidate the role of the medial basal hypothalamus (MBH) in stress-induced prolactin (Prl) release, adult male rats which had received monosodium glutamate (MSG 4 mg/g) on alternate days for the first 10 days of life were subjected to ether stress for 2 min. Blood samples were drawn at 0, 15, 30 and 60 min after etherization through an indwelling jugular catheter implanted 1 day before the experiment. Sixty minutes after etherization the dopamine receptor blocker spiroperidol (0.1 mg/kg) was injected intravenously and another blood sample was withdrawn 60 min later. Although there were no differences in basal levels of plasma Prl between control and MSG-treated animals, a dramatic elevation of plasma Prl was observed in control rats 15 min after etherization, whereas no significant change was seen in MSG-treated animals. Spiroperidol significantly increased plasma Prl in both groups of animals, but the magnitude of the increase in Prl levels of MSG-treated rats was roughly one-third (p less than 0.005) that of control animals. When morphine (3 mg/kg) was injected to test its capability to stimulate Prl release in MSG-treated rats, significant increases of similar magnitude of plasma Prl were observed in both groups. These results show that (1) the tuberoinfundibular dopaminergic system is functioning in MSG-treated rats to maintain Prl at low levels, (2) the stimulatory effect of morphine on Prl release is intact in MSG-treated rats, and (3) the tuberoinfundibular dopaminergic system is not responsible for stress-induced Prl release, but rather another mechanism located within the arcuate nucleus (ARC) and highly sensitive to destruction by MSG mediates stress-induced Prl release. PMID- 6297691 TI - Connections of the mesencephalic locomotor region (MLR) III. Intracellular recordings. AB - The responses of neurons in the area of the cat mesencephalic locomotor region (MLR) following stimulation of the entopeduncular nucleus (EN) were recorded intracellularly. At the end of each experiment a precollicular-postmamillary brainstem transection was performed and stimulation of the recording site(s) was employed to induce locomotion on a treadmill. This procedure was assumed to establish that intracellularly studied cells in the vicinity of a locomotion inducing site were MLR neurons. About 10% of MLR neurons were found to respond to stimulation of the EN at short latencies. Stimulation of MLR efferent pathways was used to identify output neurons by antidromic activation. Very few MLR output neurons were found to receive EN projections (i.e. to respond at short latency following EN stimulation). These experiments support previous results describing a sparse projection from the EN to the MLR. This projection appears to be functionally varied (EPSP, IPSP and EPSP-IPSP responses were observed in MLR neurons following EN stimulation) and to exert its major influence on interneurons, not on output neurons, of the MLR. PMID- 6297692 TI - [Focal nodular hyperplasia of the liver and hepatocellular carcinoma of the liver in children with Fanconi's anemia after long-term treatment with androgens]. PMID- 6297693 TI - [Production of suppressor factor(s) in the in vitro antibody response by periodate oxidized spleen cells]. AB - Sodium periodate treatment of Mouse spleen cells activated suppressor cells which act on the in vitro antibody response towards sheep erythrocytes or trinitrophenyl-polyacrylamide beads. Present study shows that suppression can be mediated by soluble factor (s) released in the cultures of periodate treated cells. Production of such suppressor factor (s) was associated to the presence of free aldehyde groups generated by periodate on the lymphocyte membrane. Control or cells successively treated by periodate and then reduced by borohydride were unable to produce suppressor factor (s). Suppressor factor (s) were stable at 56 degrees C (30 mn) and inactivated at 80 degrees C (10 mn). PMID- 6297694 TI - [Method for measuring the orientation rate of flowing erythrocytes by spin labeling]. AB - The electron spin resonance spectra of erythrocytes labeled by 5-nitroxide stearate, flowing through a quartz sample cell of rectangular cross-section, were studied. The main cross-sectional dimension of the quartz cell is either parallel or perpendicular to the magnetic field. Comparison between experimental spectra and numerically simulated spectra allows to determine an orientation rate of the red blood cells, as a function of the flowing velocity and of other hydrodynamic parameters. PMID- 6297695 TI - Lung cancer: current concepts and prospects. AB - In the United States, lung cancer is now the leading cancer killer among men, and it will soon be the leader among women. Most lung cancers are attributable to cigarette smoking. Symptomatic lung cancer tends to be advanced and unresectable. Chest roentgenography and sputum cytology are the only techniques proved capable of detecting presymptomatic, early-stage disease. Treatment of lung cancer depends primarily on the cell type and the extent of the tumor. Small cell cancer tends to be widespread, and the favored treatment is combination chemotherapy. For non-small cell cancer, the treatment of choice, whenever possible, is surgical resection. Efforts directed toward early detection and treatment have failed to reduce lung cancer mortality substantially. Thus, primary prevention through control of cigarette smoking remains the single most important measure for combating this frustrating disease. PMID- 6297696 TI - Metastatic chemodectoma with multiple nodular lesions. PMID- 6297697 TI - The effect of naloxone and morphine on convulsions in mice following withdrawal from nitrous oxide. AB - The incidence of withdrawal convulsions was determined in mice following removal from a 70 per cent nitrous oxide environment. Groups of 20 mice received saline (control), naloxone or morphine subcutaneous injections five minutes prior to withdrawal. The observer was blind to the treatments. In comparison to the control group, the proportion convulsing was significantly (p less than 0.05) increased following naloxone 0.125 mg (n = 40), 0.25 mg, but not 0.5 mg. The proportion convulsing was significantly decreased following morphine 0.4 mg. Overall proportions of mice convulsing was 0.55 for the saline control group; 0.73 for naloxone 0.125 mg; 0.80 for naloxone 0.25 mg; 0.60 for naloxone 0.50 mg; and 0.38 for morphine 0.4 mg. Modification of this phenomenon by both an opiate antagonist and agonist suggests endorphin withdrawal as a possible mechanism. However, this should be regarded as indirect evidence pending further study of this area. PMID- 6297698 TI - Intraoperative pulmonary tumor embolism after hepatectomy for liver carcinoma. AB - Intraoperative pulmonary tumor embolism occurred during right hepatectomy for primary hepatic carcinoma. The tumor embolus produced abrupt arterial hypotension, tachycardia, an increased central venous pressure, intense cyanosis, followed by cardiac arrest. Pulmonary tumor embolism resulting from hepatic tumors is rare and has not been reported to have occurred intraoperatively. When tumor thrombosis has been detected or suspected, hepatic vascular exclusion should be considered for the prevention of intraoperative embolism. PMID- 6297699 TI - Femoral neuropathy: a complication of lithotomy position under spinal anaesthesia. Report of three cases. AB - Three patients developed solitary unilateral peripheral femoral neuropathy after vaginal hysterectomy. All were operated under subarachnoid analgesia in the lithotomy position. Straight rod leg supports with swing stirrups were used and the procedures lasted for two and one-half hours. The complication is thought due to the extreme abduction of thighs with external rotation at the hip causing ischaemia of the femoral nerve as it is kinked beneath the tough inguinal ligament. The prognosis was found to be excellent with complete recovery within eight to ten weeks. The complication is preventable by using lateral thigh supports limiting the degree of abduction. PMID- 6297700 TI - Admission of whole families. AB - This is a program description of a short-term residential unit in which whole families are assessed and treated. This unit provides an alternative treatment format for exploring situations where removal of the child appears to be the only remaining option. It also provides an in-depth consultation for agencies who are "stuck" in their treatment of family systems and allows for the teaching of family systems intervention techniques in a non-threatening way. It also allows an opportunity for an external system to reframe, what are, to the involved clinician, impasses with clinical problems. The program, because of its structure, introduces anxiety and tension into systems with chronic mental health problems. It, therefore, serves as, not a panacea, but as an adjunct to more traditional treatment for rigid and resistant systems. PMID- 6297701 TI - Hyperthermic isolation-perfusion in vivo of the canine liver. PMID- 6297702 TI - Coexisting lobular neoplasia and carcinoma of the breast. AB - In a review of 3040 cases of carcinoma of the breast of all types in the files of the Laboratory of Surgical Pathology at Columbia for the years 1960 to 1980, 267 cases were found in which the lobular neoplasia lesion coexisted with one of the usual forms of breast carcinoma. These patients had a separate and distinct, and of course malignant, clinicopathologic entity which is distinguished from benign lobular neoplasia occurring alone. Comparing these findings in lobular neoplasia coexisting with one of the usual forms of carcinoma with our findings in lobular neoplasia occurring alone, it was found that the patients with the latter lesion were younger. Three of the nine microscopic features studied in both forms of lobular proliferation were considerably more frequent in lobular neoplasia coexisting with carcinoma: (1) loss of cohesion of the cells filling up the lobules; (2) macroacini; and (3) a maximal amount of lobular neoplasia. The great majority of the forms of carcinoma that were found coexisting with lobular neoplasia were well differentiated, small cell, intraductal, and tubular, and metastasized less often than carcinomas usually do. Carcinoma developed in the second breast three times more frequently in patients with lobular neoplasia preceding or coexisting with unilateral carcinoma than it did in patients without lobular neoplasia. PMID- 6297703 TI - Spontaneous rupture of primary hepatoma: report of 63 cases with particular reference to the pathogenesis and rationale treatment by hepatic artery ligation. AB - Of 508 cases of primary liver cancer admitted during 1970-1978, 63 cases had spontaneous rupture of carcinomatous nodules. The association with cirrhosis among noncomplicated cases was 65% and 93% in cases with the rupture (P less than 0.001). In most instances, the presenting symptoms were not dramatic and could be indistinguishable from other uncomplicated carcinoma of the liver. Sixteen patients were treated conservatively and all died. The other ten patients who presumably bled due to coagulation defect were also given conservative treatment and the mortality rate was only 60%. Hepatic artery ligation was employed in 26 cases with 54% mortality but bleeding stopped in 92%, as compared with other conventional surgical measures such as packing, suture, and cauterization to control hemorrhage with 90% mortality rate. There is ample evidence that hepatic venous obstruction due to the tumor invasion and portal hypertension due to the pre-existing cirrhosis play important roles in the pathogenesis of spontaneous rupture of the tumor. Hepatic artery ligation is a rationale treatment because it not only cuts off blood supply to the tumor nodule which is almost exclusively nourished by arterial blood, but also reduces the degree of portal hypertension in cirrhosis. PMID- 6297704 TI - Cytogenetic analysis of collagenase- releasing rabbit VX-2 carcinoma-derived cells. AB - Primary cultures of rabbit VX-2 carcinoma appeared to be heterogeneous, containing at least two morphologically distinct cell types, fibroblast-like cells (F cells) and epithelioid cells (E cells). Ultrastructural studies suggested that the E cells were of epithelial origin, whereas the F cells did not differ significantly from normal rabbit fibroblasts. Cytogenetic studies showed that the E cells were characterized by numerical and structural chromosomal changes which remained constant subsequent to serial in vitro culture and in vivo transplants in rabbits. Analysis of G-banded chromosomes from E cells revealed a modal number of 54 with relatively few normal chromosomes and a variety of distinctive, mostly unidentifiable marker chromosomes. The origin of only four marker chromosomes could be partially determined. The ratio of normal to marker chromosomes remained relatively constant following serial transplants in rabbits. The F cells appeared to be derived mainly from host tissue, as they contained a normal diploid complement with sex chromosomes corresponding to the sex of the host. PMID- 6297705 TI - Fatal hypereosinophilia with chromosome 15q- in a patient with multiple primary and familial neoplasms. AB - A man with large-cell carcinoma of the lung, cerebral meningioma, occult adenocarcinoma of the prostate, and follicular adenoma of the thyroid developed symptomatic, rapidly progressive hypereosinophilia with abnormalities of eosinophil ultrastructure and bone marrow karyotype (45,X,15q22-). Although the patient's eosinophilia defied strict classification as idiopathic hypereosinophilic syndrome (HES), simple tumor-associated eosinophilia, or eosinophilic leukemia, it appeared to be incited by the lung cancer and quickly acquired malignant independence. The family had an excess of prostate cancer and lymphoproliferative neoplasms. PMID- 6297706 TI - Cytogenetic analysis of SV40-transformed human breast epithelial cells. AB - The SV40-transformed breast epithelial cell lines established by Chang et al. [1] were shown to be hypotetraploid and characterized by six chromosome markers: M1 i(1q), M2 del(1)(q21), M3 i(6p), M4 del(1)(q11), M5 t(8p;12q), and M6 dir dup(11)(p12 leads to pter). The presence of common chromosome markers indicates that these cell lines are probably derived from the same original transformed cell. PMID- 6297707 TI - Chromosome translocations, surface immunoglobulins, and Epstein-Barr virus in Japanese Burkitt's lymphoma. PMID- 6297708 TI - Regional localization by light microscopic autoradiography of receptors in mouse brain for phorbol ester tumor promoters. AB - The receptors for the phorbol ester tumor promoters are present in highest concentration in the brain. We have examined the distribution of receptors in mouse brain by light microscopic autoradiography. Marked differences in receptor levels were found both between brain regions and between layers of a single region. Binding was high in hippocampus, cerebral cortex, striatum and substantia nigra, lower in thalamus and pons. In cerebellum, binding was high in the stratum moleculare, intermediate in the stratum granulosum and low in the lamina alba. The localization suggests a functional role for the receptors within brain, and it permits comparison with other known activities. For example, the pattern of localization is distinct from those reported for neurotransmitter receptors or for phospholipids. PMID- 6297709 TI - Epstein-Barr virus activation by tung oil, extracts of Aleurites fordii and its diterpene ester 12-O-hexadecanoyl-16-hydroxyphorbol-13-acetate. AB - During the screening of plant oils for their Epstein-Barr virus (EBV)-activating potency, we found that tung oil possesses an activity comparable to croton oil. Tung oil from various sources and the extracts from its parental plant Aleurites fordii (Chinese tung oil tree), when used in combination with n-butyrate, were shown to efficiently activate EBV persisting in human lymphoblastoid Raji cells (non-producer). The major diterpene ester in the plant extract, 12-O-hexadecanoyl 16-hydroxyphorbol-13-acetate (HHPA), also exerted a similar activity. In producer P3HR-1 cells, both tung oil and HHPA increased the yield of infectious EBV by approximately five-fold. Since tung oil is used for the manufacture of oil paints, varnishes, waterproof substance, anticorrosives and other products, the implication of using such an agent with EBV-activating potency in our daily life is assessed and discussed. PMID- 6297710 TI - An overview of current concepts in the management of patients with testicular tumors of germ cell origin--Part I: Pathophysiology, diagnosis, and staging. PMID- 6297711 TI - Effects of antibodies to choriogonadotropin in malignant growth. II. Solid transplantable rat tumors. AB - We have studied the effects of preimmunization with a conjugate of beta-subunit of choriogonadotropin and tetanus toxoid (CG beta-tt) on the growth of the implanted R 3230 AC mammary adenocarcinoma (in Fischer 344 rats) and the implanted 5123 1-1 hepatoma (in Buffalo rats) after 20 days, in order to determine if the in vivo production of antibodies against CG could modify the relationship between host and malignant growth. The results obtained demonstrated that active immunization against CG retarded significantly (P less than 0.01) the growth of the two transplantable tumors. Anti-CG antibodies were also determined and constantly found in the sera of all the preimmunized rats of both strains while no antibodies to CG were found in the control animals. PMID- 6297712 TI - Hepatitis A virus infection of HBsAg-producing hepatocellular carcinomas in athymic mice. PMID- 6297713 TI - A quantitative and qualitative study of blood monocytes in patients with bronchogenic carcinoma. AB - Absolute circulating number and functions of blood monocytes (i.e., pinocytosis, phagocytosis, and chemotaxis) were studied in 25 patients with untreated bronchogenic carcinoma and in 28 control subjects. The absolute circulating monocyte count was increased in 20 (80%) of the patients. There was no difference in the pinocytic and phagocytic activity of patient and control monocytes. In contrast, patient monocytes showed depressed chemotactic responsiveness. This defect was more severe in small cell anaplastic carcinoma than in the other histologic types of bronchogenic carcinoma (P = 0.0001), and may explain the difference in macrophage infiltration seen in solid tumours of the lung. There was no correlation between chemotaxis and clinical stage. Depressed chemotaxis may be related to a plasma factor, since patient plasma inhibited the chemotaxis of control monocytes as well as the activity of chemotactic agents. The defective chemotaxis and the presence of plasma inhibitory activity may interfere with the ability of blood monocytes to accumulate as macrophages in tumour sites. PMID- 6297714 TI - Kinetics of the reactivity of subpopulations of spleen cells of mice bearing virus-induced mammary tumors to syngeneic antigenic extracts in vitro. Supportive and suppressive effects of macrophages. AB - This study was designed to investigate the nature of lymphocyte reactivity to soluble tumor antigens with respect to the kinetics of the reactivity, the responding cell type, and the role of accessory cells, within a syngeneic system. BALB/c mice were inoculated with 1 X 10(6) viable cells of syngeneic MTV-induced mammary tumors. Assessment of proliferative activity of spleen cells of these animals by DNA synthesis (3H-thymidine incorporation in vitro) indicated a biphasic response to stimulation by 200 micrograms of a syngeneic perchloric acid (PCA)-soluble extract (AMMT) of the tumor over a 25-day period, with peak activities at days 13 and 19 post inoculation. The response was predominantly T cell-mediated. Splenic macrophage population rose from less than 2% of total spleen cells by day 25 without any appreciable change in the T or B cell population. Depletion of spleen cells of macrophages abolished the first peak activity (at day 13) but significantly enhanced the second (at day 19). Reconstitution of the depleted cells with macrophages prepared from peritoneal exudates of tumor-bearing or normal mice restored the responses to undepleted values, thus indicating an accessory role for macrophages in these responses. These results provide new data which should contribute to a better understanding of the tumor-host relationship. PMID- 6297715 TI - Normalization of defective monocyte chemotaxis during chemotherapy in patients with small cell anaplastic carcinoma of the lung. AB - Monocyte chemotactic responsiveness (MCR) in 14 patients with small cell anaplastic bronchogenic carcinoma was depressed before treatment compared with the MCR in 28 normal controls (P = 0.00004). MCR was subsequently monitored during combination chemotherapy and after 6 months the MCR had become normalized compared with pretreatment values (P = 0.00006). In addition, chemotactic factor inhibitor (CFI) activity in plasma was measured before treatment and after 6 months. When incubated with plasma before treatment casein had 62% of normal activity and when incubated with plasma after chemotherapy, 81% of normal activity (P = 0.0009). CFI activity decreased by greater amounts in patients in complete remission than in patients in partial remission or in non-responders (P = 0.01). This study supports the concept that cancer patients have depressed monocyte function. Chemotherapy seems to enhance monocyte chemotaxis in vitro and to decrease CFI activity in plasma. PMID- 6297716 TI - Formation of benzo(a)pyrene/DNA adducts and their relationship to tumor initiation in mouse epidermis. AB - The tumorigenicity of benzo(a)pyrene [B(a)P] applied topically as a skin tumor initiator in Sencar mice and the formation of epidermal B(a)P/deoxyribonucleoside adducts were compared over a similar range of doses (50 to 1600 nmol). The tumor initiating activity of B(a)P, its covalent binding to mouse epidermal DNA, and the formation of the major hydrocarbon/deoxyribonucleoside adduct showed approximately parallel dose-response curves. The major hydrocarbon/deoxyribonucleoside adduct formed cochromatographed with marker adducts of (N2-(10S-[7R,8S,9R-trihydroxy-7,8,9,10-tetrahydrobenzo(a)pyrene]y) deoxyguanosine while other minor adducts also were observed. The disappearance of DNA-bound products in the epidermis was followed for 21 days after an initiating dose of B(a)P (100 nmol) was applied topically to the mice. The half-lives of the B(a)P/deoxyribonucleoside adducts and the total radioactivity bound to the DNA were 4.5 and 5.5 days, respectively. However, in spite of the loss of measurable DNA-bound material, the tumor yield was unchanged regardless of whether promotion was begun 7 or 21 days after initiation. The results suggest a possible causal relationship between B(a)P/deoxyribonucleoside adduct formation and papilloma formation in mouse skin. PMID- 6297717 TI - Nuclear thyroid hormone receptors in C3H/HeN mouse mammary glands and spontaneous tumors. AB - Saturable, high-affinity binding sites for 3,5,3'-triiodo-L-thyronine (T3) were identified in isolated nuclei and solubilized chromatin extracts of mammary glands, spontaneous mammary tumors, and liver from C3H/HeN mice. Receptor concentration in whole mammary gland nuclei (254 fmol/mg DNA) was only about one half that of mouse liver nuclei (536 fmol/mg DNA), but in molecular weight (55,000) and in their affinity for various thyroid hormone analogues, the binding was essentially identical. Saturation analysis of T3 binding in a series of individual spontaneous mammary tumors and pooled lactating mammary glands indicated that the concentrations of T3-binding sites of the mammary gland are conserved in the transition to neoplasms and are somewhat increased in the largest tumors. Thyroxine binding was identical in capacity to T3 binding in mammary gland nuclei and nuclear extract but showed a higher binding capacity than did T3 in the largest tumors. High-performance molecular exclusion chromatography did show a difference between mammary gland and liver in the distribution of competible [125I]T3 binding between two macromolecular forms; the excluded peak (Mr greater than 450,000) comprised 56% of the T3 binding in the liver but only 9% in the mammary gland with the included peak (Mr 55,000) contributing the balance of binding in each case. Spontaneous mammary tumor resembled the mammary gland in the macromolecular distribution of specific T3 binding (16% excluded). Thymidine uptake showed only a modest decrease in the larger tumors (greater than 2.0 g), while nuclear histone acetylase activity was significantly decreased in this group. Neither measurement showed a significant correlation with T3 or thyroxine binding capacity. PMID- 6297718 TI - Tumor promoter binding to rat mammary cell cultures: role of receptors in the inhibition of dome formation. AB - We have studied the receptors for phorbol ester tumor promoters in a set of cultures of mammary cell lines derived from the same dimethylbenz(a)anthracene induced tumor. These lines differ in cell morphology and in the ability to form domes either spontaneously or under the action of inducers. Tumor promoters inhibit dome formation. We asked whether receptors for tumor promoters could also mediate dome induction. We were unable to detect high-affinity receptors for the strong promoter, tetradecanoyl-12-phorbol-13-acetate (TPA) but could reveal receptors for the weaker promoter, phorbol-12, 13-dibutyrate (PDBU). Receptors were easily demonstrable during PDBU binding at 4 degrees; at 37 degrees, there was rapid disappearance of bound ligand, accompanied by its breakdown. Most cell lines have two classes of receptors with different affinities. Receptors for TPA could be demonstrated by competition with PDBU binding. In LA7 cells, prolonged exposure to PDBU caused down-regulation of the PDBU receptors with lower affinity. The half-effective doses of the dome-inhibiting effects are comparable to the KdS of the higher-affinity PDBU receptors measured by binding. TPA binds preferentially to the same class of receptors. This class of receptors, therefore, appears to be the one mostly involved in the anti-dome effect of either PDBU or TPA. Because dome inducers do not compete with PDBU binding, the PDBU receptors may not be responsible for dome induction. PMID- 6297719 TI - Cyclic adenosine 3':5'-monophosphate-dependent and -independent protein kinases in human leukemic cells. AB - The pattern of protein kinase activity in leukemic cells from patients with chronic myelocytic leukemia, acute myeloblastic leukemia, acute monocytic leukemia, chronic lymphocytic leukemia, and acute lymphoblastic leukemia was studied and compared with normal peripheral blood granulocytes and lymphocytes. Our data showed that: (a) histone kinase activity was slightly lower in leukemic cells than in normal cells, whereas casein kinase activity was 2- to 3-fold higher in leukemic cells; (b) cyclic adenosine 3':5'-monophosphate stimulated 1.4 to 1.6-fold histone kinase activity of both normal and leukemic cells, whereas it did not stimulate casein kinase activity; (c) the ratio of histone kinase activities to casein kinase activities correlated directly with the maturation of the white blood cells; and (d) histone and casein kinase activities of extracts from normal and leukemic cells behaved similarly on chromatography on phosphocellulose and casein/Sepharose 4B. These results suggest that the increase in casein kinase activity is not due to the appearance of a new type of casein kinase but to an increase of the casein kinases 1 and 2 present in normal cells. PMID- 6297720 TI - Invasive properties of primary pediatric neoplasms in vitro. AB - Primary solid tumors were mechanically and/or enzymatically disassociated, and the resulting suspensions of single cells and small clumps of cells were seeded onto three different substrates, i.e., tissue culture plastic, rat smooth muscle cells (SMCs), and SMC-derived extracellular matrix. Tests of the relative effectiveness of these substrates in supporting the survival and/or growth of ten different neoplasms demonstrated that only two explants remained viable for longer than 2 weeks when seeded onto tissue culture plastic while nine of the ten survived on biological substrates for 1 month or longer. Thus, tissue culture plastic was a poor substrate for primary pediatric neoplasms. In general, more than 80% of the most common solid neoplasms in childhood (brain tumor, neuroblastoma, renal tumor, rhabdomyosarcoma, osteogenic sarcoma, and Ewing's sarcoma) routinely survived or grew in long-term cultures when cultured onto SMCs or their matrix. Both substrates were effective in promoting survival and/or growth; however, cells of neuroblastomas and certain brain tumors showed a preference for a living smooth muscle substrate. Tumor cells maintained their characteristic cellular and subcellular morphology when compared with the histology of the in vivo neoplastic lesions. Light and electron microscopy of selected neoplasms cultured on SMCs for various time periods demonstrated areas of distinct cellular invasion and/or partial destruction of the SMC multilayers which correlated with the invasive potentials of the neoplasms in patients. Invasion and destruction of the SMCs were also noticed with quiescent tumor cell cultures, indicating that growth was not a necessary property of invasion. Several neoplasms were also capable of the degradation of connective tissue proteins as indicated by the hydrolysis of radiolabeled SMC matrices, but simple correlations between the extent of matrix degradation and invasive ability could not be drawn. The culture system described consistently provided for the survival and/or growth of the most common pediatric tumors for long time periods. Thus, basic biological properties of primary tumors, e.g., growth, invasive potentials, and differentiation capabilities, could be investigated routinely. PMID- 6297721 TI - Characterization of Epstein-Barr virus-carrying cell lines established from chronic lymphocytic leukemia. AB - Attempts were made to establish lymphoid cell lines from the cultured peripheral blood lymphocytes of six patients with chronic lymphocytic leukemia. In only one case was cell growth obtained following the addition of exogenous transforming Epstein-Barr virus, and those cell cultures proved not to have acquired the ability to proliferate permanently. In the same case, cell lines were established spontaneously from the peripheral blood without addition of Epstein-Barr virus. The cells which grew spontaneously were large, were occasionally weakly surface adherent, and grew in suspension as loose clumps or as single cells. They were negative for surface immunoglobulins and spontaneous rosette formation with sheep erythrocytes and positive for intracytoplasmic immunoglobulins (Fc and C3 receptors). At an early passage, the spontaneous lines had an aneuploid karyotype with some triploid and some tetraploid cells. Structural chromosomal aberrations include a 14q+. Electron microscopy of the chronic lymphocytic leukemia lines revealed relatively smooth surfaces with numerous mitochondria, widespread vacuolization, and numerous unusual "myelin" figures. Five to 10% of the cells were phagocytic as detected by internalization of latex particles; however, they were Epstein-Barr nuclear antigen positive. The nature of these cells and their possible relationship to the etiology of chronic lymphocytic leukemia are discussed. PMID- 6297722 TI - Influence of asbestos on the uptake of benzo(a)pyrene and DNA alkylation in hamster tracheal epithelial cells. AB - The objective of these experiments was to understand the mechanism of cocarcinogenicity of asbestos and polycyclic aromatic hydrocarbons. Benzo(a)pyrene [B(a)P] was coated onto crocidolite or chrysotile asbestos fibers, resuspended in serum-free medium, and added to cultures of hamster tracheal epithelial cells. The fibers markedly enhanced cell uptake of B(a)P. Although considerable metabolism occurred, approximately 40% of the applied B(a)P was retained by the cells after 8-hr incubation as opposed to 5% after incubation with B(a)P in the absence of asbestos. The hydrocarbon-containing medium was replaced by fresh medium. Four days later, approximately 3% of the B(a)P that had been applied when adsorbed to asbestos was still persistent in cells as compared to 0.5% in cells treated with B(a)P alone. DNA from hamster tracheal epithelial cells was purified, and the amount of B(a)P alkylation was assessed. At 8 hr, the extent of alkylation after treatment of the cells with either B(a)P or B(a)P:asbestos was similar. However, the retained unmetabolized B(a)P was subsequently metabolized and contributed to further alkylation so that the B(a)P asbestos treated cells demonstrated considerably higher levels of alkylation throughout the 4-day posttreatment period. None of these effects was observed if asbestos was added 1 hr before the addition of B(a)P. The enhanced uptake of B(a)P and subsequent additional alkylation of DNA might represent a mechanism of asbestos-induced cocarcinogenesis. PMID- 6297723 TI - Differential effect of coformycin on the cell cycle traverse of normal and simian virus 40-transformed human fibroblasts. AB - Normal and simian virus 40-transformed human embryonic lung fibroblasts in culture were exposed to 3.5 microM coformycin (CF). This treatment resulted in almost complete inhibition of adenosine deaminase activity in both cell lines and retarded the progress of the fibroblasts through the cell cycle, as measured by the rate of cell proliferation. There was a marked difference, however, in the effect of CF on the traverse of different segments of the cell cycle. In normal fibroblasts, CF rapidly but transiently inhibited cell entry into visible stages of mitosis and delayed the progress through S and G2. In transformed fibroblasts, the effects of CF on the cell cycle included an early acceleration of the cell entry into visible mitosis and prolongation of this phase of the cell cycle. These results indicate that inhibition of adenosine deaminase can have different effects on the traverse of the critical segments of the cell cycle in normal and transformed cells. PMID- 6297724 TI - Relationship of surface membranes of lymphoid cells transformed by Abelson murine leukemia virus to tumor rejection. PMID- 6297725 TI - Metabolism of benzo(a)pyrene and 1-naphthol in cultured human tumorous and nontumorous colon. AB - The oxidative metabolism of benzo(a)pyrene and the conjugative metabolism of 1 naphthol by explant cultures of normal human colon and colonic tumor tissue, obtained at surgery, have been studied. After 24 hr in culture, the explants were exposed to either [1-14C]-1-naphthol (20 to 100 microM) or [3H]-benzo(a) pyrene (1.5 microM) for a further 1.5 to 24 hr. Both normal-appearing tissue and tumor tissue metabolized benzo(a)pyrene to a wide variety of organic solvent-soluble metabolites, including monohydroxybenzo(a)pyrenes, dihydrodiols, and tetrols. 1 Naphthol was metabolized by cultured human colonic mucosa and tumor tissue to both its glucuronic acid and sulfate ester conjugates. In the normal tissues, with naphthol (20 microM), sulfate ester conjugation predominated. However, with the tumor tissue, sulfate ester conjugation decreased; thus, the percentage of glucuronic acid conjugates, expressed as a percentage of total metabolites formed, was increased significantly compared to normal tissue. The relationship, if any, of these changes to neoplastic transformation is unclear. The technique of explant culture described in this study may be of use for the study of other facets of the pathobiology of solid tumors. PMID- 6297726 TI - Multiple forms of protein kinase from normal human brain and glioblastoma. AB - The biochemical characteristics of the protein kinase (PK; adenosine triphosphate protein phosphotransferase, EC 2.7.1.37) isozymes in subcellular preparations from normal human brain cortex and glioblastoma were investigated after chromatography on diethylaminoethyl cellulose, and the following results have been obtained. Two major isozyme forms, eluted by 50 and 200 mM phosphate buffer, are present in both cytosol and membrane-derived preparations from cerebral cortex. Furthermore, these isozyme forms have properties similar to those referred to as type I and type II cyclic adenosine 3':5'-monophosphate-dependent PK. In these chromatographic isozymes, cyclic adenosine 3';5'-monophosphate is more active in stimulating the basal PK enzyme than is cyclic guanosine 3':5' monophosphate. In glioblastoma, the PK activity from cytosol and particulate preparations is resolved by diethylaminoethyl cellulose in four peaks. In cytosol, the major portion of the enzyme is eluted with a 300 mM buffer (about 50% of the total basal PK activity) and is cyclic nucleotide dependent. On the contrary, in glioblastoma particulate, the PK enzyme is mainly eluted at 50 and 100 mM buffer; neither of these isozymes is cyclic nucleotide dependent. As for cytosol, only the particulate isozyme eluted at 300 mM buffer is strongly activated by cyclic nucleotides. Finally, in both glioblastoma subcellular preparations, only a type II cyclic adenosine 3':5'-monophosphate-dependent PK is present. PMID- 6297727 TI - Human papilloma virus 5-DNA in a carcinoma of an epidermodysplasia verruciformis patient infected with various human papillomavirus types. AB - An epidermodysplasia verruciformis patient suffering from generalized warts and a carcinoma at the forehead was found to be infected by at least six types and subtypes of human papillomaviruses. The central part of the carcinoma, however, harbored only human papillomavirus 5 DNA. The DNA persisted extrachromosomally in high genome copy number. In contrast to wart DNA preparations, a significant part of the viral sequences in the carcinoma was present as oligomers, at least part of them being concatemers as shown by S1 digest. The human papillomavirus 5 subtype from this carcinoma was compared with the two other carcinoma isolates described so far and proved to be rather similar. PMID- 6297728 TI - Mutagenesis in Chinese hamster cells by cyclopenta(a)phenanthrenes activated by a human hepatoma cell line. AB - The cyclopenta(a)phenanthrene, 15,16-dihydro-11-methyl-cyclopenta(a)phenanthren 17-one, had potent mutagenic activity in cell-mediated mutation assays with V79 Chinese hamster cells as targets, and cells of the human hepatoma line HepG2 as mediators of activation. The compound was inactive when low-passage hamster embryo cells were used as activators. When the mutagenic activity of a series of cyclopenta(a)phenanthrenes was compared in mutation assays with HepG2 cells as activators, there was a good correlation between mutagenic activity in this system and carcinogenic activity in mouse skin in vivo. One exception was a noncarcinogenic compound, which is mutagenic in the Ames' test, and was also mutagenic in the mammalian cell assay. PMID- 6297729 TI - Inhibition by lithium of PGE1-sensitive adenylate cyclase in neuroblastoma X glioma hybrid cells: approach to the attenuation of the opiate withdrawal syndrome. AB - Exposure of neuroblastoma X glioma hybrid cells to LiCl (Li+) for 48 h significantly reduces prostaglandin E1 (PGE1)-dependent cyclic adenosine monophosphate (AMP) accumulation. When applied simultaneously with morphine for the same period, Li+ inhibits the excessive accumulation of cyclic AMP in response to PGE1 characteristically exhibited by these cells upon opiate withdrawal. Likewise, Li+ inhibits, in vitro, PGE1-dependent cyclic AMP accumulation in tissue slices from rat corpus striatum. Prophylactic administration of Li+ in the diet attenuates the withdrawal symptoms in morphine addicted rats. These findings support clinical reports that suggest that prophylactic treatment with Li+ attenuates the severity of the opiate withdrawal syndrome. PMID- 6297731 TI - Small cell carcinoma of the lung. PMID- 6297730 TI - Phase II clinical study of Yoshi 864 in squamous cell carcinoma of the uterine cervix. PMID- 6297732 TI - Spontaneous arteriovenous fistula between common iliac artery and vein. PMID- 6297733 TI - Experimental studies on new liquid embolization mixtures (histoacryl-lipiodol, histoacryl-panthopaque). AB - In vitro measurements have proved that the polymerization time of n butylcyanoacrylate (Histoacryl) can be delayed markedly and controlled by adding an oily radiopaque material. Our animal experiments have shown that a mixture of Histoacryl and Lipiodol ultrafluid at a 1:3 ratio provided an optimal embolization material with good flow properties, polymerization time of approximately 7 sec and excellent contrast definition. This mixture may be handled easily, with a minimum of risk involved when used with a 40% glucose solution. PMID- 6297734 TI - 2',3'-Cyclic nucleotide 3'-phosphodiesterase (CNPase) activity in cultured nerve cell lines from central nervous system: comparison of proliferating and resting growth states and cell cycle-dependent activity changes. AB - 1. The present communication is concerned with the expression and cell cycle dependent regulation of the enzyme 2',3'-cyclic nucleotide 3'-phosphodiesterase (CNPase) in cultured nerve cell lines derived from the rat central nervous system (CNS). 2. The enzyme activity was measured in relation to two reversible serum controlled growth states (exponentially growing/quiescent) including a comparison of the enzyme activities in cell lines of neuronal and glial origin as well as in fibroblasts. CNPase is present in all cell types tested, but the enzyme activity is very sensitive to changes in the cellular growth state. Nerve cell lines in exponentially growing cultures express a 3 to 15 times higher specific CNPase activity than the nonneural cell types. In serum-starved quiescent cultures, the differences in specific enzyme activity between the nerve cell lines and the fibroblasts are enlarged even more up to a ratio of about 50 to 150, indicating a specific function of this enzyme within the central nervous system. 3. Neuron like B104 cells could be stimulated to synchronized growth by serum readdition to quiescent cultures. A series of ordered activity changes of CNPase has been observed after the reinitiation of cell growth. The enzyme is stimulated at two particular stages during the cell cycle, leading to a biphasic activity profile. Maximum stimulation of CNPase correlates with the G1 phase. 4. Hydroxyurea induced blockage of synchronized B104 cells to traverse the S phase also prevents the subsequent stimulation of CNPase activity. Therefore, we conclude that a correlation exists between the periodic activity changes of CNPase and particular phases of the B104 cell cycle. PMID- 6297735 TI - Activity and immunocytochemical localization of 2',3'-cyclic nucleotide 3' phosphodiesterase (CNPase) in primary nerve cell cultures from rat brain. PMID- 6297736 TI - Cytochemical demonstration of an enzymatic production of hydrogen peroxide on the surface of isolated thyroid capillaries. AB - By means of a cytochemical technique, hydrogen peroxide formation was located on the endothelial cell surface (predominantly the luminal aspect) of capillaries obtained by collagenase digestion of rat thyroid. The cyanide-insensitive H2O2 formation required aerobic conditions and NAD(P)H as substrate. FAD could also stimulate the reaction, but not xanthine. The cytochemical reaction was blocked by a non-penetrating protein inhibitor. The observations are interpreted as evidence of a plasmalemma-bound H2O2-generating enzyme. The findings indicate that microvascular endothelial cells are involved in the release of activated oxygen species, which might have important pathophysiologic implications. PMID- 6297737 TI - Stimulated inositol lipid metabolism: an introduction. PMID- 6297739 TI - Phosphatidylinositol metabolism in the adrenal medulla. AB - Changes in phosphoinositide metabolism due to muscarinic stimulation of the adrenal medulla are reviewed. Evidence is presented that muscarinic receptors inhibit catecholamine secretion by the bovine gland and that muscarinic agonists do not cause entry of calcium ions. Results are inconsistent with the theory that phosphatidylinositol hydrolysis opens calcium 'gates'. Polyphosphoinositide metabolism is also reviewed and the suggestion made that phosphatidylinositol 4,5 bisphosphate may regulate the activity of the calcium pump ATPase in cells where phosphoinositide-linked receptors promote calcium influx. PMID- 6297738 TI - The enzymology of stimulated inositol lipid turnover. PMID- 6297740 TI - Inositol lipids and cell stimulation in mammalian salivary gland. AB - The rat parotid salivary gland shows marked alterations in phospholipid metabolism when stimulated by certain agonists. These agonists are those which cause cellular Ca mobilization by activation of muscarinic, alpha-adrenergic or peptidergic (substance P) receptors. The phospholipid changes apparently reflect the activation of a phosphoinositide-phosphatidic acid cycle, the precise pathways of which are not known with certainty. The observed effects include (1) an increased labelling by 32PO4 of phosphatidylinositol and phosphatidic acid, (2) net synthesis of phosphatidic acid, (3) net breakdown of phosphatidylinositol and phosphatidylinositol-4,5-bisphosphate. These effects apparently do not require the presence of extracellular Ca or the release of internal Ca and cannot be produced by the artificial introduction of Ca into the cytosol with Ca ionophores. These findings are consistent with the view that a receptor-mediated alteration in phosphoinositide metabolism represents an early step in the stimulus-response pathway in the parotid acinar cell. It has been suggested that phosphatidic acid synthesis might be of central importance in mediating Ca influx and that PIP2 breakdown might play a role in activation of Ca release. Evidence for these latter ideas is for the present largely circumstantial. PMID- 6297741 TI - Ligand-stimulated inositol lipid metabolism in the liver: relationship to receptor function. PMID- 6297742 TI - The role of inositide phospholipids in the action of steroidogenic hormones. PMID- 6297743 TI - Phosphatidylinositol 4-phosphate and phosphatidylinositol 4,5-bisphosphate: lipids in search of a function. PMID- 6297744 TI - Substitution of 5' helper virus sequences into non-rel portion of reticuloendotheliosis virus strain T suppresses transformation of chicken spleen cells. AB - The genome of the highly oncogenic avian retrovirus reticuloendotheliosis virus strain T (REV-T) differs from that of the helper virus reticuloendotheliosis virus strain A by a substitution (rel and a large deletion. Further deletions, constructed in vitro, within the helper-virus-related sequences of REV-T have little effect on the ability of the virus to transform chicken spleen cells in vitro. However, deletions that extend into rel abolish transformation. Substitution of helper-virus-related sequences for the deleted region in the non rel portion of REV-T also abolishes transformation. Viruses with revertant phenotype were obtained both spontaneously and by construction in vitro from these substituted recombinants. The revertant viruses have various mutations, including deletions and insertions, in the helper-virus-related sequences. Thus the additional helper-virus-related sequences suppress expression of transformation in cis, and the deletion in REV-T seems necessary for expression of the transforming properties of the virus. PMID- 6297745 TI - Self-splicing RNA: autoexcision and autocyclization of the ribosomal RNA intervening sequence of Tetrahymena. AB - In the macronuclear rRNA genes of Tetrahymena thermophila, a 413 bp intervening sequence (IVS) interrupts the 26S rRNA-coding region. A restriction fragment of the rDNA containing the IVS and portions of the adjacent rRNA sequences (exons) was inserted downstream from the lac UV5 promoter in a recombinant plasmid. Transcription of this template by purified Escherichia coli RNA polymerase in vitro produced a shortened version of the pre-rRNA, which was then deproteinized. When incubated with monovalent and divalent cations and a guanosine factor, this RNA underwent splicing. The reactions that were characterized included the precise excision of the IVS, attachment of guanosine to the 5' end of the IVS, covalent cyclization of the IVS and ligation of the exons. We conclude that splicing activity is intrinsic to the structure of the RNA, and that enzymes, small nuclear RNAs and folding of the pre-rRNA into an RNP are unnecessary for these reactions. We propose that the IVS portion of the RNA has several enzyme like properties that enable it to break and reform phosphodiester bonds. The finding of autocatalytic rearrangements of RNA molecules has implications for the mechanism and the evolution of other reactions that involve RNA. PMID- 6297746 TI - Expression of a chicken lysozyme recombinant gene is regulated by progesterone and dexamethasone after microinjection into oviduct cells. AB - We transferred a chicken lysozyme gene recombinant by microinjection into primary cultures of chicken oviduct cells. The recombinant gene is a fusion between the lysozyme promoter, including 1.4 kb of upstream sequences, and the coding region of the gene for SV40 T antigen (plys-T). The expression of plys-T is stimulated by the steroid hormones progesterone and dexamethasone, but not by estradiol. The number of oviduct cells expressing coinjected or separately injected control genes is not increased by steroids. A deletion mutant lacking the lysozyme sequences between -161 and +15 does not express T antigen, indicating that transcription of plys-T starts within the lysozyme promoter region. By screening different cell types we found that microinjected plys-T is expressed in chicken oviduct cells but not in chicken macrophages or fibroblasts or in rat II fibroblasts. PMID- 6297747 TI - Homothallic switching of yeast mating type cassettes is initiated by a double stranded cut in the MAT locus. AB - A double-stranded DNA cut has been observed in the mating type (MAT) locus of the yeast Saccharomyces cerevisiae in cultures undergoing homothallic cassette switching. Cutting is observed in exponentially growing cells of genotype HO HML alpha MAT alpha HMR alpha or HO HMLa MATa HMRa, which switch continuously, but not in a/alpha HO/HO diploid strains, in which homothallic switching is known to be shut off. Stationary phase cultures do not exhibit the cut. Although this site specific cut occurs in a sequence (Z1) common to the silent HML and HMR cassettes and to MAT, only the Z1 sequence at the MAT locus is cut. The cut at MAT occurs in the absence of the HML and HMR donor cassettes, suggesting that cutting initiates the switching process. An assay for switching on hybrid plasmids containing mata- cassettes has been devised, and deletion mapping has shown that the cut site is required for efficient switching. Thus a double-stranded cut at the MAT locus appears to initiate cassette transposition-substitution and defines MAT as the recipient in this process. PMID- 6297748 TI - Yeast DNA replication in vitro: initiation and elongation events mimic in vivo processes. AB - An enzyme system prepared from Saccharomyces cerevisiae carries out the replication of exogenous yeast plasmid DNA. Replication in vitro mimics that in vivo in that DNA synthesis in extracts of strain cdc8, a temperature-sensitive DNA replication mutant, is thermolabile relative to the wild-type, and in that aphidicolin inhibits replication in vitro. Furthermore, only plasmids containing a functional yeast replicator, ARS, initiate replication at a specific site in vitro. Analysis of replicative intermediates shows that plasmid YRp7, which contains the chromosomal replicator ARS1, initiates bidirectional replication in a 100 bp region within the sequence required for autonomous replication in vivo. Plasmids containing ARS2, another chromosomal replicator, and the ARS region of the endogenous yeast plasmid 2 microns circle give similar results, suggesting that ARS sequences are specific origins of chromosomal replication. Used in conjunction with deletion mapping, the in vitro system allows definition of the minimal sequences required for the initiation of replication. PMID- 6297749 TI - A previously unidentified gene in the spc operon of Escherichia coli K12 specifies a component of the protein export machinery. AB - The gene prlA codes for a factor that appears to function in the export of proteins in Escherichia coli. This conclusion is based on the finding that mutations altering the prlA gene product restore export of envelope proteins with defective signal sequences. Previous results showed that the prlA gene lies in an operon (spc) known to code for ten different ribosomal proteins. Our studies show that the prlA gene lies promoter-distal to the last known ribosomal protein gene in this operon. Evidence from gene fusions constructed in vitro suggests that prlA codes for a protein containing at least 300 amino acids. Thus a heretofore unidentified protein specified by a gene within the spc operon appears to be a component of the cellular protein export machinery. PMID- 6297750 TI - Determination of the amount of homology required for recombination in bacteriophage T4. AB - Homology is an important feature of recombination. We have used the rll cistrons of bacteriophage T4 to determine the extent of homology required for recombination. We varied the amount of homologous DNA available for recombination in both marker rescue experiments and deletion-by-deletion crosses. Our results suggest that the primary pathway for recombination in T4 requires 50 bp of homology. Our finding that recombination is detectable when fewer than 50 bp of homology are available suggests that there is a second, less efficient pathway of recombination in T4. This pathway may be used during the formation of deletions. PMID- 6297751 TI - Escherichia coli DNA topoisomerase I mutants: increased supercoiling is corrected by mutations near gyrase genes. AB - Bacterial chromosomes and plasmid (pBR322) DNA from topoisomerase I-defective Escherichia coli strains have been characterized with respect to superhelical density. The topoisomerase I defect results in increased negative superhelical density of both the bacterial chromosome and pBR322. Thus topoisomerase I is involved in determining the level of supercoiling in bacteria. Three of the topoisomerase I-defective strains were studied carry secondary mutations that decrease superhelical density; these additional mutations are closely linked to the gyrB locus in two of the strains and to the gyrA locus in the third strain. PMID- 6297752 TI - Escherichia coli DNA topoisomerase I mutants have compensatory mutations in DNA gyrase genes. AB - Escherichia coli deletion mutants lacking DNA topoisomerase I have been identified previously and shown to grow at a normal rate. We show that such strains grow normally only because of spontaneously arising mutations that compensate for the topoisomerase I defect. Several of these compensatory mutations have been found to map at or near the genes encoding DNA gyrase, gyrA and gyrB. DNA gyrase assays of crude extracts show that strains carrying the mutations have lower gyrase activity. Thus the mutations are in the gyrase structural genes or in nearby regulatory sequences. These results, in conjunction with DNA supercoiling measurements of others, indicate that in vivo DNA superhelicity is a result of a balance between topoisomerase I and gyrase activities. An excess of negative supercoils due to an absence of topoisomerase I is deleterious to the cell, but a moderate gyrase deficiency is not harmful. PMID- 6297753 TI - Homology between mammalian cell DNA sequences and human herpesvirus genomes detected by a hybridization procedure with high-complexity probe. AB - Mouse and human DNA used as in vitro-labeled "high-complexity" probes revealed hybridization between specific herpesvirus DNA fragments on Southern transfers and repetitive sequences present at 10(3) to 10(5) copies per mammalian cell genome. Several different sites of major cell-virus sequence homology have been detected in both the herpes simplex virus type 1 and type 2 genomes, and these are located predominantly within the L and S inverted repeat regions and near the center of the L unique region. The hybrids persisted even in relatively stringent conditions, and appear to correlate closely with some of the previously recognized regions of size heterogeneity in the viral genome. Cloned viral DNA fragments from each site hybridized to different sets of discrete bands and dispersed elements within restriction-endonuclease-digested genomic DNA from a variety of vertebrate species. Localized cell-virus homology was also detected in both the human Epstein-Barr virus and cytomegalovirus genomes. PMID- 6297754 TI - Structural and functional homologies in the receptors for insulin and the insulin like growth factors. PMID- 6297755 TI - Homology between murine and human cellular DNA sequences and the terminal repetition of the S component of herpes simplex virus type 1 DNA. AB - DNA from trigeminal ganglia of mice latently infected with herpes simplex virus type 1 was cloned in bacteriophage lambda Charon 27. Three recombinant clones that cross-hybridize to each other and hybridize to the S-segment terminal repeat region of the herpesvirus genome were obtained. The region of homology maps within a 1.1 kb Sma l fragment approximately 400 bp from the S-segment terminal repeat terminus, outside the domain of the a sequences. The insert DNAs in these clones, as well as the Sma l fragment of viral DNA that contains the region of homology, hybridize to uninfected mouse brain DNA with a pattern indicative of the presence of repeated sequences, and also hybridize to discrete Eco Rl fragments of human DNA. These recombinant clones contain intermediate repetitive mouse genomic DNA sequences related to a small region of the viral S-segment terminal repeat, and those sequences are evolutionarily conserved in mammalian DNA. PMID- 6297756 TI - Structure and role of the herpes simplex virus DNA termini in inversion, circularization and generation of virion DNA. AB - The herpes simplex virus genome consists of two components, L and S, which invert relative to each other during viral replication. The a sequence is present at the genomic termini in direct orientation and at the L-S junction in inverted orientation. Previously, we showed that insertion of a fragment spanning the L-S junction into the viral genome causes additional inversions. In this study, we determine the nucleotide sequence of the genomic termini and show that insertion of either the free S terminus or the L terminus causes inversions in the viral genome. We conclude that the a sequence is the inversion-specific sequence, that linear unit-length molecules packaged in virions are generated by cleavage between adjacent copies of the a sequence, that cleavage produces 3' single-base extensions on the genomic termini and that the signal for cleavage is contained within the a sequence. PMID- 6297757 TI - Many tumors induced by the mouse mammary tumor virus contain a provirus integrated in the same region of the host genome. AB - We have asked whether oncogenesis by the mouse mammary tumor virus (MMTV), a slowly oncogenic retrovirus, involves integration of viral DNA within a certain region of the host genome. We first identified a C3H mouse mammary tumor bearing a single new MMTV provirus and cloned a 19 kilobase (kb) DNA restriction fragment containing a junction of viral and host sequences. Host sequences from this clone were used to retrieve 25 kb of the uninterrupted locus (termed MMTV int1) from a bacteriophage library of normal mouse DNA. Hybridization with subcloned DNA fragments of MMTV int1 detected abnormal restriction fragments in digests of DNA from 18 of 26 C3H mammary tumors. The rearrangements all appeared to be due to the insertion of an MMTV provirus, and the integration sites were located in at least five clusters over a total distance of 19 kb. A polyadenylated 2.6 kb RNA species transcribed from int1 was found in the few tumors tested, but not in lactating mammary glands from C3H mice. Of 12 tested viral oncogenes, none exhibited homology with cloned DNA from this locus. We propose that tumorigenesis by MMTV is strongly favored by proviral insertion within the int1 locus, perhaps as a consequence of enhanced expression of a novel cellular oncogene. PMID- 6297758 TI - Tn5 transposition and its regulation. PMID- 6297759 TI - Nucleotide sequence of yeast LEU2 shows 5'-noncoding region has sequences cognate to leucine. AB - The LEU2 structural gene and its regulatory sequences were isolated on a 2200 bp Xho I-Sal I fragment. Sequencing of the 5'-noncoding region showed that at -151 there is an open reading frame of 23 codons of which six are for leucine. The leucine codon usage in this reading frame follows exactly that of other yeast genes. At the carboxy-terminal end and immediately after the peptide reading frame, a 14 bp hairpin (followed by a T-rich segment) can form in the putative mRNA; this arrangement closely resembles an RNA polymerase terminator. These and other features suggest a model for regulation. Preceding this is a gene (which starts at -463) for tRNALeu3, the major tRNALeu isoacceptor. RNA polymerase III transcription start and termination signals flank 5' and 3' ends, respectively, of the structural gene. The features noted above are in the same DNA strand that codes for the LEU2 gene product. PMID- 6297760 TI - Identification of base mismatches recognized by the heteroduplex-DNA-repair system of Streptococcus pneumoniae. AB - The susceptibility to repair of particular base mismatches by the hex system of Streptococcus pneumoniae was examined by comparison of the nucleotide sequence of the wild-type and eight mutant alleles of the malM gene. A detailed restriction map was constructed for pLS70, and the nucleotide sequence was determined for its 3475 bp chromosomal insert, which contains the entire malM gene (encoding amylomaltase), portions of malX and malP (encoding a membrane protein and a phosphorylase, respectively) and a control region. Transition mismatches were highly susceptible to repair; transversion mismatches, much less so. A mismatch caused by a single-nucleotide deletion was reparable, but mismatches with longer deletions were not. The hex system also reduced spontaneous reversion of mutations corresponding to transitions. It is suggested that recognition of donor or nascent DNA strands by the hex system depends on single-strand breaks in the target strand, and that the role of DNA methylation in mismatch repair of Escherichia coli can be accommodated to this model. PMID- 6297761 TI - The structure of the thymidine kinase gene promoter: nuclease hypersensitivity correlates with expression. AB - Accurate and quantitative transcription of the tk gene requires sequence elements that reside within 110 nucleotides of 5'-flanking DNA. We have determined the boundaries of a hypersensitive region in 5'DNA flanking the tk gene by analyzing the relative sensitivity of specific restriction sites clustered in this region. Five different restriction enzymes, recognizing over 50 sites in the promoter region of the tk gene, each show a preferential cleavage in nuclei at a restricted number of sites residing between positions -4 and -182. Thus the tk promoter sequences are contained entirely within a hypersensitive region. Analysis of this site in tk+ transformants, a tk- mutant and tk+ rerevertant indicates that expression of tk RNA is correlated with structural alterations in the tk promoter. PMID- 6297762 TI - Functional relationships between transcriptional control signals of the thymidine kinase gene of herpes simplex virus. AB - Transcriptional control signals occur at three separate locations upstream from the herpes virus thymidine kinase gene. I have used two approaches to examine how these signals function in relation to one another. First, double mutants that simultaneously mutate various pairs of transcriptional signals were constructed. Analyses of the transcriptional phenotypes of such mutants suggest that the two most distally located signals may function in the same metabolic step, whereas the proximal signal appears to function in a process distinct from that of the distal signals. Second, the distances that normally separate the three transcriptional signals were systematically altered. These condensation and expansion mutants were studied to determine to what extent the spatial relationship between the signals is important to their function. The transcriptional phenotypes of these spacing-change mutants show that the amount of DNA that separates the three transcriptional signals is not rigidly fixed. PMID- 6297763 TI - Cell-type-specific actin mRNA populations in dictyostelium discoideum. AB - We have analyzed actin mRNA sequences present in the terminal stages of the development of Dictyostelium discoideum. Four different actin mRNA sequences were detected in migrating pseudoplasmodia. Nucleotide sequence analysis of primer extension products derived from the four mRNA sequences showed that they each encoded an actin protein with the same eight N-terminal amino acids and that they did not derive from transcription of any previously characterized actin gene. Preculmination pseudoplasmodia of Dictyostelium contain two distinct populations of committed cells, termed prespore and prestalk cells. We show that prestalk cells contain all four of the actin mRNA sequences found in pseudoplasmodia, while prespore cells contain only three of the sequences, and mature spores contain only two. Thus there is a differential loss of actin mRNA sequences during spore-cell differentiation in Dictyostelium. PMID- 6297764 TI - Sea urchin (lytechinus pictus) late-stage histone H3 and H4 genes: characterization and mapping of a clustered but nontandemly linked multigene family. AB - We have cloned and characterized members of a small multigene family that encodes late-stage histone H3 and H4 mRNAs from the sea urchin Lytechinus pictus. Unlike their highly repetitive histone gene counterparts, which are expressed at an earlier developmental stage, late H3 and H4 histone genes are not present in tandem repeats. In addition, the late stage H3 and H4 genes are not always tightly clustered together with the H1, H2A and H2B genes as they are in early histone genes. The spacer DNA that separates adjoining H3 and H4 coding regions is not conserved between nonallelic members of the late histone gene family. We have determined the nucleotide sequence of a continuous 2100 bp segment of DNA including both H3 and H4 coding sequences, the entire spacer DNA separating the genes and surrounding nonhistone DNA. The late histone H3 and H4 genes encode proteins identical to their early gene counterparts; however, the 5' leader sequence is shorter in late genes and the codon usage is different. PMID- 6297765 TI - Contact points between a positive transcription factor and the Xenopus 5S RNA gene. AB - We have investigated the contact points of a positive transcription factor with the internal control region of the 5S ribosomal RNA genes of Xenopus. The methylation of any one of eight G residues clustered at the 3' end of the internal control region on the noncoding strand of the DNA or the ethylation of their surrounding phosphates interferes with the binding of this protein. The importance of the 5' half of the control region is shown by a binding affinity of the protein to oocyte 5S RNA genes that is one fourth that to somatic 5S RNA genes. This can be attributed to two base changes at residues 53 and 55 in the 5' part of the control region. The different affinities of the transcription factor for the oocyte and somatic 5S RNA genes may contribute to their differential expression in somatic cells. The almost exclusive location of strong contact points on the noncoding strand of the gene suggests how transcription events could proceed repeatedly along a gene that is assembled into a stable transcription complex. It is proposed that the complex is transiently shifted to the noncoding strand as each round of RNA synthesis occurs from the coding strand. PMID- 6297766 TI - Nucleotide sequence of the retroviral leukemia gene v-myb and its cellular progenitor c-myb: the architecture of a transduced oncogene. AB - The oncogene (v-myb) of avian myeloblastosis virus and a large portion of its cellular homolog (c-myb) have been molecularly cloned and sequenced. The portion of c-myb we analyzed contains seven interspersed segments (or exons). Fusion of these exons creates a continuous nucleotide sequence that is remarkably similar to the sequence of v-myb and that potentially encodes a protein very similar to that specified by v-myb. Comparisons between the sequences of v-myb and c-myb indicate that transduction of c-myb to form v-myb probably resulted from an initial DNA rearrangement and the subsequent use of a spliced RNA as an intermediate. PMID- 6297767 TI - The transforming proteins of Rous sarcoma virus, Harvey sarcoma virus and Abelson virus contain tightly bound lipid. AB - We have found that the transforming proteins of Rous sarcoma virus, Harvey sarcoma virus and Abelson virus all contain tightly bound lipid. This modification could play a role in the binding of these proteins to cellular membranes. The lipid associated with p60src, the transforming protein of Rous sarcoma virus, is located in the NH2-terminal domain of the polypeptide. This is the region of the protein that has been shown previously to participate in binding the protein to membranes. Two mature forms of p21, the transforming protein of Harvey sarcoma virus, contain lipid. Lipid is not, however, associated with newly synthesized p21. While mature p60src and p21 are bound to cellular membranes, the newly synthesized forms of these proteins are not. The posttranslational addition of lipid may therefore be the means by which these proteins acquire an affinity for membranes. PMID- 6297768 TI - Specific binding of the glucocorticoid-receptor complex to the mouse mammary tumor proviral promoter region. AB - To elucidate the molecular mechanism by which steroid hormones exert their regulatory function, we investigated the interaction of a glucocorticoid-receptor complex with purified DNA fragments from cloned mouse mammary tumor (MMTV) proviral DNA. With a DNA-cellulose binding assay, rat and mouse glucocorticoid receptors were found to interact with a high affinity site or sites in or near the promoter region of the MMTV proviral DNA. The assay allowed the use of unpurified as well as purified receptor, and therefore made it possible to investigate the binding properties of mutant receptors. Two nuclear-transfer deficient receptors have a decreased affinity for specific as well as unspecific DNA, but are still capable of distinguishing between the two types of DNA. The existence of specific DNA binding sites for steroid-receptor complexes is discussed in the context of a general model of steroid hormone action. PMID- 6297769 TI - Replication origins in the Xenopus egg. PMID- 6297770 TI - beta-Endorphin. PMID- 6297771 TI - Disproportionate expression of the two nonallelic rat insulin genes in a pancreatic tumor is due to translational control. AB - The expression of the two nonallelic but highly homologous rat insulin genes (1 and 2) in a transplantable beta-cell tumor is found to be 10-fold higher for rat1 insulin than rat2 insulin, while in normal pancreatic tissue there are approximately equal amounts of each protein. No large sequence rearrangements of the genes were apparent by restriction analysis of the tumor DNA, and both genes were found to be specifically hypomethylated in the tumor as compared with other nonpancreatic tissue. Equivalent amounts of both insulin 1 and 2 precursor transcripts, as well as stable, mature mRNAs were detected in the tumor. However, two-dimensional gel analysis of immunoprecipitated rat1 and rat2 preproinsulins synthesized in vitro revealed a 10:1 ratio of rat1 to rat2 proteins. A 1:1 ratio was obtained when the tumor mRNA was treated in vitro with vaccinia virus capping extract, suggesting a structural modification at the 5' terminus of the rat2 mRNA. These results are discussed in the context of insulin regulation by glucose, shown to be due to translational control. PMID- 6297772 TI - Adenovirus VAI RNA is required for efficient translation of viral mRNAs at late times after infection. AB - Two adenovirus type 5 mutants were constructed to probe the function of the virus encoded RNA polymerase III transcripts (VA RNAs). Each mutant fails to synthesize one of the two VA RNA species. The variant that does not produce the minor VAII species grows normally. The mutant that cannot synthesize the major VAI species grows more poorly than its parent. Analysis of the mutant's growth defect indicates that the adenovirus VAI RNA is required for efficient translation of viral mRNAs at late times after infection. PMID- 6297773 TI - The structure of the human zeta-globin gene and a closely linked, nearly identical pseudogene. AB - DNA sequencing studies indicate that only one of two closely linked human embryonic alpha-like globin genes, zeta (zeta), encodes a functional polypeptide. The other is a pseudogene (psi zeta) that differs by only 3 bp in the protein coding sequence, one of which converts the codon for amino acid 6 into a chain termination codon. Both zeta-globin genes differ from all other alpha-like genes thus far reported in that they contain large introns consisting, in part, of simple repeat sequences. Intron 1 of each gene contains a variation of the repeat sequence ACAGTGGGGAGGGG, while intron 2 contains the repeat sequence CGGGG. Comparison of the human zeta- and alpha-globin gene sequences reveals that the embryonic and adult alpha-like genes began to diverge from each other relatively early in vertebrate evolution (400 million years ago). In contrast, the beta-like embryonic globin gene, epsilon (epsilon), is the product of a much more recent evolutionary event (200 million years ago). Thus, even though the temporal and quantitative expression of zeta- and epsilon-globin genes must be coordinately controlled during development, their evolutionary histories are clearly distinct. PMID- 6297774 TI - Molecular cloning of sequences from a Drosophila RNA polymerase II locus by P element transposon tagging. AB - We have identified a lethal mutation in the D. melanogaster RNA polymerase II locus, RpIIC4, caused by insertion of a transposable element associated with the phenomenon of hybrid dysgenesis (P element). Using previously cloned P element sequences as a hybridization probe we have isolated a hybrid lambda phage clone carrying a 10 kb genomic DNA fragment containing a 1.3 kb P element insert and flanking sequences from the RpII locus. The non-P sequences in this clone (lambda DmRpII-1) hybridize to polytene chromosome band region 10C, the cytogenetic location of RpIIC4, and revertants which lose the lethal RNA polymerase II mutation also lose P element sequences from the locus. We have generated several additional P element insertions into the locus and shown that they can occur at two or more different sites. These experiments illustrate that mutagenesis by P element insertion and use of cloned P DNA to retrieve the DNA sequences into which insertion has occurred may be a general method for cloning genetically defined loci in Drosophila. PMID- 6297775 TI - Variation in hormone autonomy and regenerative potential of cells transformed by strain A66 of Agrobacterium tumefaciens. AB - Mutant Agrobacterium tumefaciens strain A66 is shown to differ from its wild-type progenitor (strain A6) by a spontaneous 2.7 kb DNA insert into the T-DNA region of its Ti plasmid. Tobacco stems transformed by A66 exhibit an attenuated response characterized by slow growth and shoot proliferation. Clonal analysis demonstrates that this response is due to an alteration in the growth and regenerative potential of transformed cells, rather than to variation in the frequency of fully autonomous cells within the primary tumor. Cloned A66 transformed tobacco cells exhibit an auxin requirement for growth that can be overcome by shoot proliferation. Other host species, however, may complement the A66 mutation yielding fully auxin-independent tumors when transformed by this bacterium. PMID- 6297776 TI - Purification of an adenovirus-coded DNA polymerase that is required for initiation of DNA replication. AB - Temperature-sensitive mutants in the N complementation group of human adenovirus type 5 are defective at the nonpermissive temperature for replication of virus DNA and for transformation of rat embryo cells. We show that nuclear extracts prepared from Ad5ts 149-infected cells grown at the nonpermissive temperature fail to replicate DNA in vitro. The defect lies in the first step in the initiation of viral DNA synthesis, the formation of a covalent linkage between the terminal protein precursor (pTP) and dCMP. A 140 kilodalton (140 kd) protein which complements these defective extracts and contains DNA polymerase activity has been purified from HeLa cells infected with wild-type Ad2. It is tightly associated with the 80 kd pTP in a replication complex. Both of these proteins are products of the E2B region of the adenovirus genome, and the 140 kd protein coding sequences lie immediately downstream from those encoding the 80 kd protein. These results demonstrate that adenovirus encodes a novel DNA polymerase that is required for priming of DNA synthesis at the origin of replication. This protein may also function in the initiation of transformation of cultured cells. PMID- 6297777 TI - Reconstitution studies detect a single polymerase entry site on the vesicular stomatitis virus genome. AB - To identify the initial steps of vesicular stomatitis virus transcription, we reconstituted purified nucleocapsid template with solubilized transcriptase and characterized the in vitro products of de novo transcription. In the absence of UTP and GTP, only leader gene products were synthesized; mRNA oligonucleotides were detected only after transcription of full-length leader was permitted. These data suggest that vesicular stomatitis virus polymerase does not enter the genome independently at each gene, but each polymerase begins transcription at the 3' end of the genome, and reaches internal genes only by sequentially transcribing the 3' preceding sequences. These results are consistent with the conclusion that the observed sequential transcription of vesicular stomatitis virus mRNAs is due to obligatory entrance of all polymerases at the leader gene, and suggest that the transcriptase and replicase may recognize the same promoter. PMID- 6297778 TI - Transformation of both erythroid and myeloid cells by E26, an avian leukemia virus that contains the myb gene. AB - E26 and avian myeloblastosis virus are replication-defective avian retroviruses that contain the myb oncogene and cause leukemia in chickens with short periods of latency. Animals infected with E26 develop erythroleukemia and also contain low numbers of transformed myeloid cells, while avian myeloblastosis virus induces a purely myeloid leukemia. In both cases the type of leukemia induced is independent of the subgroup of the helper virus used. E26-transformed erythroid and myeloid cells can each be propagated selectively from explanted leukemic cells with media supplemented with factors that promote the growth either of normal chicken erythroid precursors or of myeloid progenitor cells. E26 also induces the outgrowth of transformed cells from bone marrow cells infected in vitro. These cells are also either erythroid or myeloid, depending on the culture conditions employed. Most of the erythroid cells transformed by E26 are erythroblast-like, but a significant number are more mature, including erythrocyte-like cells as well as some cells that appear to be aberrant in differentiation. Both erythroid and myeloid E26-transformed cells produce infectious virus and express P135 E26, the putative (gag-myb-x) transforming protein of the virus. Thus E26 is a virus that is capable of generating factor dependent transformed cells in two different hematopoietic lineages. PMID- 6297780 TI - Asynchronous switching of flagellar motors on a single bacterial cell. AB - Salmonella possesses several flagella, each capable of counterclockwise and clockwise rotation. Counterclockwise rotation produces swimming, clockwise rotation produces tumbling. Switching between senses occurs stochastically. The rotational sense of individual flagella on a single cell could be monitored under special conditions (partially de-energized cells of cheC and cheZ mutants). Switching was totally asynchronous, indicating that the stochastic process operates at the level of the individual organelle. Coordinated rotation in the flagellar bundle during swimming may therefore derive simply from a high counterclockwise probability enhanced by mechanical interactions, and not from a synchronizing switch mechanism. Different flagella on a given cell had different switching probabilities, on a time scale (greater than 2 min) spanning many switching events. This heterogeneity may reflect permanent structural differences, or slow fluctuations in some regulatory process. PMID- 6297779 TI - A UDP-glucose:glycoprotein glucose-1-phosphotransferase in embryonic chicken neural retina. AB - A subclass of cell-surface glycoproteins from embryonic chicken neural retina contains a high mannose-type oligosaccharide that terminates with glucose linked via a phosphodiester bond to penultimate mannose. This unusual oligosaccharide seems responsible for the glycoprotein attachments to the cell-surface baseplate ligatin. Using beta-32P-UDP-3H-glucose, we demonstrate in retinal homogenates the existence of a UDP-glucose:glycoprotein glucose-1-phosphotransferase (GlcPTase) that catalyzes the synthesis of such a linkage. Characterization of the doubly labeled product resulting from activity of the transferase reveals a family of endoglycosidase H-sensitive oligosaccharides displaying a cation-exchange profile similar to that of oligosaccharides derived from ligatin-associated proteins synthesized in vivo. Further analysis confirms that the incorporation of label is due to a terminal 3H-glucose joined via a 32P-phosphodiester linkage to carbon 6 of a penultimate mannose. We propose that GlcPTase may be a controlling enzyme for the targeting of certain newly synthesized proteins to the cell surface. PMID- 6297781 TI - The replication initiator protein of plasmid R6K tagged with beta-galactosidase shows sequence-specific DNA-binding. AB - We have tagged the replication initiator protein of the plasmid R6K near the C terminal end by fusion, in the correct reading frame, with the 89 amino acid long N-terminal alpha-donor polypeptide of beta-galactosidase of E. coli. This fusion was carried out with recombinant DNA methods. The protein chimera thus generated retained the activities of both initiation of DNA replication in vivo at the replication origin gamma of R6K and hydrolysis of beta-galactopyranoside when complemented in vivo with the alpha-acceptor polypeptide coded by the lac Z gene containing the M15 deletion. Using the simple and convenient assay for detecting beta-galactosidase, we have partially purified the tagged replication initiator, and have demonstrated that the protein binds to specific DNA sequences of the R6K chromosome. The protein bound to DNA sequences located at two places in the 5' untranslated leader region of the initiator protein cistron. PMID- 6297782 TI - Autoregulation of the Escherichia coli crp gene: CRP is a transcriptional repressor for its own gene. AB - The restriction fragments carrying the region preceding the Escherichia coli crp structural gene were transcribed. The 5' end of the crp mRNA was determined by RNAase partial digestion and S1 digestion methods. Thus the crp gene has been shown to possess a 167 bp leader. CRP-cAMP specifically prevents the crp transcription. In other words, the crp gene is regulated autogenously. DNAase foot-printing studies indicated that CRP-cAMP binds to the crp gene at positions +26 to +67. This region exhibits a striking sequence homology to the CRP-binding sites in other genes. CRP and RNA polymerase bind to the crp regulatory region simultaneously. These results suggest a different mechanism for transcriptional repression of the crp gene by CRP-cAMP from that of a typical operator-repressor model. PMID- 6297783 TI - Role of the Xis protein of bacteriophage lambda in a specific reactive complex at the attR prophage attachment site. AB - Phage lambda controls its integration and excision by differential catalysis of the forward and reverse reactions. The lambda Int protein is required for both directions, but Xis for excision only. Previous electron microscopic observations have shown that Int protein forms a stable, condensed protein-DNA complex with the phage (attP) and prophage left (attL) substrate sites, but not with the host (attB) or prophage right (attR) sites. We have found that Int and Xis together produce a stable, condensed complex with attR. The attR complex involves the P region DNA to the left of the crossover point (O site). In contrast, the attP complex includes DNA on both sides of the crossover point (P and P'), and the attL structure involves the P' DNA to the right of O. In the presence of Int and Xis, the attL and attR sites form a paired structure. We conclude that the role of Xis is to provide a distinct reactive structure at attR, allowing attL and attR to pair efficiently. PMID- 6297784 TI - Transforming capacities of avian erythroblastosis virus mutants deleted in the erbA or erbB oncogenes. AB - Mutants of avian erythroblastosis virus (AEV) were constructed by deleting large nucleotide segments in each of the viral oncogenes termed v-erbA and v-erbB. Mutants in erbA (erbA -B +) retained the ability to transform fibroblasts in vitro, and these cells exhibited most of the transformation characteristics that typify wild-type AEV-transformed fibroblasts. In addition, the mutants induced small erythroid colonies upon infection of bone marrow cells in culture. Chickens inoculated with erbA -B + virus or with erbA -B +-transformed cells developed sarcomas or atypical erythroid leukemias. The erythroid cells transformed in vivo or in vitro by the erbA -B + viruses appeared not to be as tightly blocked in differentiation as wild-type transformed cells. In contrast, fibroblasts infected with the erbA +B - mutant resembled normal cells in all transformation parameters tested, and no bone marrow cell transformation was observed with the mutant. The results indicate that the main transforming properties of AEV are encoded in erbB and that its effects are enhanced by erbA. PMID- 6297786 TI - Transposition of Tn3 and related transposons. PMID- 6297785 TI - The asialoglycoprotein receptor internalizes and recycles independently of the transferrin and insulin receptors. AB - Receptor-mediated endocytosis of specific ligands is mediated through clustering of receptor-ligand complexes in coated pits on the cell surface, followed by internalization of the complex into endocytic vesicles. We show that internalization of asialoglycoprotein by HepG2 hepatoma cells is accompanied by a rapid (t1/2 = 0.5-1 min) depletion of surface asialoglycoprotein receptors. This is followed by a rapid (t1/2 = 2-4 min) reappearance of surface receptors; most of these originate from endocytosed cell-surface receptors. The loss and reappearance of asialoglycoprotein receptors is specific, and depends on prebinding of ligand to its receptor. HepG2 cells also contain abundant receptors for both insulin and transferrin. Endocytosis of asialoglycoprotein and its receptor has no effect on the number of surface binding sites for transferrin or insulin. We conclude that binding of asialoglycoprotein to its surface receptor triggers a rapid and specific endocytosis of the receptor-ligand complex, probably due to a clustering in clathrin-coated pits or vesicles. PMID- 6297787 TI - Endogenous retroviruses. PMID- 6297788 TI - Evidence for a transposon in Caenorhabditis elegans. AB - The C. elegans genome contains a 1.7 kb repeated DNA sequence (Tc1) that is present in different numbers in various strains. In strain Bristol and 10 other strains analyzed, there are 20 +/- 5 copies of Tc1, and these are located at a nearly constant set of sites in the DNA. In Bergerac, however, there are 200 +/- 50 interspersed copies of Tc1 that have arisen by insertion of Tc1 elements into new genomic sites. The interspersed copies of Tc1 have a conserved, nonpermuted structure. The structure of genomic Tc1 elements was analyzed by the cloning of a single Tc1 element from Bergerac and the comparison of its structure with homologous genomic sequences in Bristol and Bergerac. Tc1 elements at three sites analyzed in Bergerac undergo apparently precise excision from their points of insertion at high frequency. PMID- 6297789 TI - Analysis of a yeast nuclear gene involved in the maturation of mitochondrial pre messenger RNA of the cytochrome oxidase subunit I. AB - We have analyzed the mitochondrial RNA of a yeast nuclear pet mutant with no cytochrome oxidase activity. The product of the gene affected in this mutant appears to be necessary for the correct maturation of the mitochondrial pre-mRNA of the cytochrome oxidase subunit I. It does not affect, however, the overall splicing of cytochrome b pre-mRNA or the intron excision of the 21S ribosomal RNA precursor. This gene has been isolated by genetic complementation in yeast, and its DNA sequence has been determined. It is transcribed, as detected by S1 mapping experiments, and could encode a protein of 436 amino acids. PMID- 6297790 TI - How are proteins imported into mitochondria? PMID- 6297791 TI - Partition mechanism of F plasmid: two plasmid gene-encoded products and a cis acting region are involved in partition. AB - Plasmids that replicate using the replication origin (oriC) of the E. coli chromosome are not stably inherited through cell division, but can be stabilized by joining with a particular segment of F plasmid that presumably provides the partition function. The segment necessary for stabilization has been located within a 3.0 kb segment outside of the region essential for autonomous replication of the F plasmid. This segment contains three functionally distinct regions: two of them (designated sopA and sopB) specify gene products that act in trans, whereas the third region (sopC) acts in cis. All three functions seem to be essential for normal partition of the plasmid into daughter cells during cell division. The cis-acting region also specifies plasmid incompatibility. PMID- 6297792 TI - Two intron sequences in yeast mitochondrial COX1 gene: homology among URF containing introns and strain-dependent variation in flanking exons. AB - The DNA sequences of two optional introns in the gene for subunit I of cytochome c oxidase in yeast mitochondrial DNA have been determined. Both contain long unassigned reading frames (URFs). These display regions of amino acid homology with six other URFs, two of which encode proteins involved in mitochondrial RNA splicing. Such conserved regions may thus define functionally important domains of proteins involved in RNA processing. This homology also implies that these URFs had a common ancestral sequence, which has been duplicated and dispersed around the genome. Comparison of the flanking exons in the long strain KL14-4A with their unsplit counterpart in D273-10B reveals clustered sequence differences, which lead in D273-10B to codons rarely used in exons. These differences may be linked to the loss or absence of one of the optional introns. PMID- 6297793 TI - Unequal excision of complementary strands is involved in the generation of palindromic repetitions of rho- mitochondrial DNA in yeast. AB - In the rho- mutants of yeast, the mitochondrial genome is made up of a small segment excised from the wild-type mitochondrial DNA. The segment is repeated either in tandem or in palindrome to form a series of multimeric DNAs. We have asked how the palindromic organization arises. From several palindromic rho- mitochondrial DNAs, we have isolated the restriction fragments that contained the head-to-head or tail-to-tail junction of the repeating units, and have determined their nucleotide sequences. We found that the palindromes were not symmetrical right up to the junction points: at the junction, there was always an asymmetrical sequence of variable length. At both ends of this junction sequence, we found inverted oligonucleotide sequences that were variable in each mutant and that were present in the wild-type DNA. At the moment of excision, a single strand cut seems to occur at each of these short inverted repeats, in such a way that the two complementary strands of the genome are cut unequally and the single stranded overhangs become the junction sequences between the palindromic repeating units. This scheme may account for the complex structures of many rho- mitochondrial DNAs. PMID- 6297794 TI - Initiation of meiosis in yeast mutants defective in adenylate cyclase and cyclic AMP-dependent protein kinase. AB - Control of the initiation of meiosis was examined in diploids of yeast homozygous for two temperature-sensitive mutations, cyr1 and CYR3, which are defective in adenylate cyclase and cAMP-dependent protein kinase, respectively. The cyr1 and CYR3 mutations permitted the initiation of meiosis, but resulted in the frequent production of two-spored asci at the restrictive temperature. Unlike the wild type diploid cells, the cyr1 and CYR3 homozygous diploid cells were capable of initiating meiosis even in nutrient growth media. This unique feature of the cyr1 and CYR3 mutants suggests that these mutations relate to the choice between mitotic and meiotic processes. In diploids homozygous for the bcy1 mutation that results in deficiency of the regulatory subunit of cAMP-dependent protein kinase and production of a high level of the catalytic subunit of this enzyme, no premeiotic DNA replication and commitment to intragenic recombination occurred, and no spores were formed. We conclude that the initiation of meiosis may be dependent upon the repression of cAMP production and the inactivation of cAMP dependent protein kinase. PMID- 6297795 TI - Rapid incorporation of label from ribonucleoside disphosphates into DNA by a cell free high molecular weight fraction from animal cell nuclei. AB - A readily sedimentable nuclear fraction from Chinese hamster embryo fibroblast (CHEF/18) cells catalyzes incorporation of 14C-rCDP into DNA. Data indicated that this incorporation is made possible by the conversion of rCDP into a small and functionally compartmentalized, rather than a large and freely diffusible, pool of dCTP. This catalytically active sedimentable fraction from S phase CHEF/18 cells or actively replicating calf thymus cells contains nascent and template DNA, and numerous enzymes required for DNA biosynthesis including ribonucleoside diphosphate reductase, thymidylate synthetase, dihydrofolate reductase, DNA methylase, topoisomerase and DNA polymerase. We have named this catalytically active macromolecule the replitase. The replitase fraction contained spherical particles with a diameter of approximately 24 to 30 nm and had an estimated molecular weight on the order of 5 X 10(6). PMID- 6297796 TI - The SV40 72 bp repeat preferentially potentiates transcription starting from proximal natural or substitute promoter elements. AB - Activation of gene expression by the SV40 72 bp repeat was studied at the transcriptional level by quantitative S1 nuclease mapping of total RNA isolated from Hela cells transfected with chimeric conalbumin promoter-SV40 early gene recombinants. Our results demonstrate that, irrespective of its orientation, the 72 bp repeat is a potentiator of initiation of transcription from "TATA"-box dependent and -independent "natural" or "substitute" promoter elements. In addition, we show that potential proximal promoter sequences are activated in preference to more distal ones. These results are consistent with the bidirectional entry site model for transcription activation by the 72 bp repeat. PMID- 6297797 TI - Gene conversion and polymorphism: generation of mouse immunoglobulin gamma 2a chain alleles by differential gene conversion by gamma 2b chain gene. AB - We have determined the complete nucleotide sequence of the C57BL/6 allele of the mouse immunoglobulin gamma 2a chain gene. A comparison with the BALB/c gamma 2a gene for 1912 nucleotides reveals that the two alleles exhibit extensive divergence, since there are 138 single-base-pair differences and 8 insertions or deletions. We have compared the two gamma 2a alleles with the two corresponding gamma 2b alleles, which differ in only 12 positions. It appears that among the 134 differences between the two gamma 2a alleles, 70 are at positions where gamma 2a and gamma 2b are identical in the BALB/c haplotype and 54 are at positions where gamma 2a and gamma 2b are identical in the C57BL/6 haplotype. All these results suggest that nonreciprocal gene conversion between nonallelic genes can introduce sequence homogeneity in linked genes and can generate extensive divergence and polymorphism in allelic genes. We suggest that the gamma 2a and gamma 2b gene ancestors freely diverged after duplication, and that the conversion events were promoted by a deletion shortening the distance between the two loci. PMID- 6297799 TI - N protein of vesicular stomatitis virus selectively encapsidates leader RNA in vitro. AB - The N protein of vesicular stomatitis virus, prepared in a soluble form, was found to self-assemble, and to assemble with RNAs into RNAase-resistant structures with the buoyant density of viral nucleocapsids. It selectively assembled leader RNAs over other viral transcripts. The basis for this selective encapsidation was not the relative size of the viral transcripts or the presence or absence of a 5' cap group, but was sequence-dependent. Partial-assembly experiments demonstrated that leader RNA assembly started within the first 14 nucleotides at the 5' end. Examination of known leader RNA sequences suggests that the sequence responsible for selective assembly by N protein is a five-times repeated A residue at every third position from the 5' end of the leader chain. PMID- 6297798 TI - Mechanism of action of a yeast RNA ligase in tRNA splicing. PMID- 6297800 TI - A specific viral DNA sequence is stably integrated in herpesvirus oncogenically transformed cells. PMID- 6297801 TI - Identification and characterization of the avian erythroblastosis virus erbB gene product as a membrane glycoprotein. AB - Avian erythroblastosis virus causes erythroid leukemia and sarcomas in chickens. The viral oncogene responsible for these diseases, erb, is divided into two regions known as erbA and erbB, and recent evidence suggests that it is the erbB gene that is responsible for the transforming activity. From rats bearing avian erythroblastosis virus-induced sarcomas, we have obtained antisera which are specific for the erb gene products. Using such antisera, we have been able to characterize the erbB gene product as a 68,000 molecular weight protein. Pulse chase and cell-free in vitro translation experiments show that the initial product is a 62,500 dalton protein which is initially modified to a 66,000 dalton protein, and then further modified to a 68,000 dalton form. These modifications could be shown to be associated with glycosylation and phosphorylation. Cell fractionation experiments revealed that the 66,000 and 68,000 dalton proteins were located in cell membrane fractions, and immunofluorescence results showed the erbB gene product to be expressed on the cell surface. PMID- 6297802 TI - Drosophila melanogaster DNA clones homologous to vertebrate oncogenes: evidence for a common ancestor to the src and abl cellular genes. AB - We have isolated phage clones containing the D. melanogaster sequence homologous to the v-abl oncogene, and two types of phage clones containing sequences homologous to the v-src probe. The D. melanogaster abl clone (lambda Dabl1) and one of the src clones (lambda Dsrc1) hybridize with both v-abl and v-src probes, and both map in situ to the same chromosomal position, 73B, on chromosome arm 3L. The second D. melanogaster src clone (lambda Dsrc2) does not react with the v-abl probe and hybridizes in situ to chromosomal position 64B. The hybridization pattern suggests that the src and abl cellular oncogenes have evolved from a common prototype sequence. The homologous sequences in D. melanogaster exhibit hybridization to regions in the vertebrate v-abl and v-src that are important for kinase activity and transforming potential of the viral gene products. PMID- 6297803 TI - BALB and Kirsten murine sarcoma viruses alter growth and differentiation of EGF dependent balb/c mouse epidermal keratinocyte lines. AB - Clonal BALB/c mouse epidermal keratinocyte (BALB/MK) cell lines were established in tissue culture. Despite their aneuploid nature, the lines were nontumorigenic, and retained in vitro properties similar to those of primary diploid keratinocytes. These included the constitutive expression of keratin and terminal differentiation in response to a calcium concentration greater than 1.0 mM in the medium. The cells also demonstrated an absolute requirement for nanomolar concentrations of epidermal growth factor (EGF) for their proliferation. BALB or Kirsten murine sarcoma viruses are acute transforming retroviruses, which have been shown to transform fibroblastic and hematopoietic cells. Infection of BALB/MK or its clonal sublines with either virus leads to the rapid acquisition of EGF-independent growth. The cells concomitantly lose their sensitivity to calcium-induced terminal differentiation. Thus these retroviruses can rapidly confer upon epithelial keratinocytes in culture growth properties that resemble those of malignant epidermoid carcinoma cells. PMID- 6297804 TI - Inhibition of VSV binding and infectivity by phosphatidylserine: is phosphatidylserine a VSV-binding site? AB - Recently we described a saturable, high-affinity binding site for vesicular stomatitis virus (VSV) on the surface of Vero cells that appears to mediate viral infectivity. To isolate this binding site, we have extracted Vero cells with the detergent, octyl-beta-D-glucopyranoside. The dialyzed detergent extract specifically inhibits the saturable, high-affinity binding of 35S-methionine labeled VSV to Vero cells. The inhibitory activity is resistant to protease, neuraminidase and heating to 100 degrees C. It is soluble in chloroform-methanol and inactivated by phospholipase C, suggesting that it is a phospholipid. Of various purified lipids tested, only phosphatidylserine was capable of totally inhibiting the high-affinity binding of VSV. The half-maximal inhibitory concentration for phosphatidylserine was 1 microM. Phosphatidylserine also inhibited VSV plaque formation by 80%-90%; Herpes simplex virus plaque formation was unaffected. Centrifugation and electron microscopy studies have shown that phosphatidylserine-containing liposomes bind to VSV. The finding that phosphatidylserine directly binds to VSV and inhibits VSV attachment and infectivity suggests that plasma membrane phosphatidylserine could function as a binding site or portion of a binding site for VSV. PMID- 6297805 TI - Enhancing effect of lithium and potassium ions on lectin-induced lymphocyte proliferation in aging and Down's syndrome subjects. AB - The effect of different concentrations of LiCl or KCl (0.6-20 meq/liter) on PHA stimulated lymphocytes from young, old, and Down's syndrome subjects was studied. LiCl showed a dramatic enhancing effect on [3H]thymidine incorporation induced by a suboptimal dose of PHA in old subjects and Down's syndrome patients. An increase of [3H]thymidine incorporation in human lymphocytes stimulated by a suboptimal dose of PHA was also observed with KCl. This effect was higher in old subjects than that observed in young and Down's subjects. LiCl and KCl can modulate and partially restore the derangement in early events of mitogen stimulation which seems to be present in lymphocytes from both old and Down's syndrome subjects. PMID- 6297806 TI - Suppression of cell-mediated immunity to challenge with P 815 mastocytoma in concanavalin A-treated mice. AB - C57Bl/6 (B6) mice allogeneic to the P 815 mastocytoma tumor cell line when treated with concanavalin A prior to and at frequent intervals following challenge intraperitoneally with 10(7) tumor cells showed a significant suppression of their cell-mediated immune response at 9-10 days when compared with untreated animals. Suppression of the immune response of mice syngeneic (DBA/2) or hybrid (BDF1) to the tumor was also evidenced by increased mortality rates in concanavalin A-treated animals. The suppression of cell-mediated cytotoxicity observed in B6 mice treated with concanavalin A could be reversed by pretreatment with 20 mg silica injected intraperitoneally 7 days prior to challenge. These results suggest that macrophages play a significant role in the concanavalin A-induced immune suppression observed in this in vivo tumor-host system. PMID- 6297807 TI - Regulation of bone marrow lymphocyte production. IV. Cells mediating the stimulation of marrow lymphocyte production by sheep red blood cells: studies in anti-IgM-suppressed mice, athymic mice, and silica-treated mice. AB - Some cellular requirements have been examined for the stimulation of lymphocyte production in mouse bone marrow by injected sheep red blood cells (SRBC). The increased genesis of marrow lymphocytes after a single dose of SRBC assayed radioautographically after [3H]thymidine labeling was unimpaired in the marrow of mice treated with anti-IgM antibodies from birth to eliminate B lymphocytes, and in congenitally athymic mice lacking T lymphocytes. However, pretreatment of mice with silica to depress macrophage function completely abolished the SRBC effect both on the total lymphocyte production and on the number of B and null small lymphocytes in the marrow. Comparative studies were performed on the thymus and spleen. The results demonstrate that the stimulation of marrow lymphocytes production by SRBC is mediated by a silica-sensitive mechanism, does not require B or T lymphocytes, and is independent of the humoral immune response. Thus, extrinsic agents may amplify the production of primary B cells and other lymphocytes in the bone marrow by an antigen-nonspecific mechanism, putatively mediated by macrophages. PMID- 6297809 TI - Immunoglobulin production by tonsil lymphocytes before and after Epstein-Barr virus transformation. PMID- 6297808 TI - Macrophage-dependent polyclonal activation of splenocytes by Bordetella pertussis endotoxin and its isolated polysaccharide and Lipid A regions. PMID- 6297810 TI - [Membrane transport of calcium in cardiac cells]. PMID- 6297811 TI - [Determination of levels of specific antibodies against the Epstein-Barr virus using the ELISA method in infectious mononucleosis and its diagnostic significance]. PMID- 6297813 TI - Time course of gold-induced accumulation of copper and zinc and the effects of dimercaptosuccinate and cadmium on gold metabolism in rat kidney. PMID- 6297812 TI - [A comparison of the gluconate and pyrophosphate heart scan in the diagnosis of acute transmural myocardial infarctions]. PMID- 6297814 TI - Biochemical evidence for chemical and/or topographic differences in the lipoperoxidative processes induced by CCl4 and iron. AB - Isolated rat hepatocytes, treated with CCl4 or ADP-Fe3+ complex show an enhanced lipid peroxidation and a decreased glucose 6-phosphatase activity. Lipid peroxidation is much more stimulated by ADP-Fe3+ or Fe3+ than by CCl4, when the metal and the haloalkane are used at a similar concentration. Increasing rates of lipid peroxidation in the different experimental conditions do not correlate with the degree of glucose 6-phosphatase inactivation, which is produced by CCl4 and not by a similar amount of ferric iron. In the case of iron, its intracellular concentration must be higher to give the enzyme inactivation exerted by CCl4. Higher intracellular levels of iron are reached when the metal is added to the cell suspension together with ADP. Under these conditions there is inactivation of glucose 6-phosphatase. Possible mechanisms accounting for a different enzyme sensitivity to iron and CCl4 are discussed. PMID- 6297815 TI - Cutaneous metabolism of benzo[a]pyrene: comparative studies in C57BL/6N and DBA/2N mice and neonatal Sprague--Dawley rats. PMID- 6297816 TI - Benzo[a]pyrene-collagen interactions. AB - In vitro interactions of benzo[a]pyrene (BaP) with acid-soluble type I collagen from rat tail tendon have been investigated. The fluorescence of BaP increases in the presence of collagen. Bound BaP inhibits the formation of collagen fibrils in solution. When BaP-collagen complexes are irradiated in air with UV (365 nm) light, BaP rapidly undergoes photooxidation with the further inhibition of fibril formation. Viscosity and circular dichroism (CD) studies show that neither BaP nor further UV-irradiation alters the size or helical conformation of the protein. During thermal denaturation of collagen, BaP fluorescence changes. Collagen from young rat tail tendon shows a pronounced drop at about 38 degrees C, whereas that from old rat tail tendon exhibits an increase with a plateau in the same temperature range. These anomalous changes are observed when tyrosine residues, present only in the non-helical terminal telopeptides of collagen, are excited at 275 nm, but not by direct BaP excitation at 387 nm. These findings suggest that the specific hydrophobic telopeptide region, which plays an important role in fibril formation, are affected by bound BaP. PMID- 6297817 TI - The electrochemical reduction of 1,4-bis-(2-[(2-hydroxyethyl)-amino] ethylamino) anthracenedione and daunomycin: biochemical significance in superoxide formation. PMID- 6297818 TI - Synthesis of charged amphipathic nitroxide lipid spin labels and an example of their application in membrane studies. AB - The synthesis of a series of amphipathic nitroxide lipid spin labels is reported. Thus, 12-proxylhexadecanol has been converted into the versatile fatty acid spin label 14-proxylstearic acid. This substance was used to prepare 14 proxylstearyltrimethylammonium methanesulfonate, a positively charged label, and 14-proxylstearylmethyl phosphate sodium salt, a negatively charged label. Also prepared in the doxyl series were quaternary ammonium salts derived from 16-doxyl and 7-doxylstearic acid. The positively charged and negatively charged proxyl labels were used in a preliminary experiment to investigate the role of charge in their interaction with reconstituted cytochrome oxidase. The average binding affinity of the negatively charged label is approximately 2-fold higher than that of the positively charged label at pH 7.4. At pH 5.5 the average relative affinity for negatively charged label is about 3.5-fold higher than that of positively charged label, suggesting that the ionizable group(s) on the protein can interact with the lipid headgroup. PMID- 6297819 TI - An outbreak of hand, foot, and mouth disease in Singapore. AB - Epidemiological studies were carried out during an extensive outbreak of hand, foot and mouth disease caused by coxsackievirus A 16 in Singapore in 1981. Most of the cases were concentrated in densely populated public housing estates, and the morbidity rate was highest in children under 5 years of age. The mean secondary attack rate for children under 12 years of age was 76.7% compared with an overall rate of 31.3%. The incubation period ranged from 1 to 7 days. Factors favouring the transmission of infection within a household included sharing of household and personal articles with the index case. The disease was mild and self-limiting. PMID- 6297820 TI - Inhibition of DNA synthesis, independent of DNA adduct formation, by benzo[a]pyrene diol epoxide in mammalian cells. AB - Treatment of SV40-infected CV-1 cells with the ultimate carcinogen anti benzo[a]pyrene diol epoxide (BPDE) at 1 X 10(-4) mg/ml or higher reduced the rate of viral DNA synthesis to an extent dependent on the BPDE concentration; similar reductions in cellular DNA synthesis were produced in infected and uninfected cells. Treatment of cells with BPDE, followed by removal of BPDE, at various times before infection with SV40 gave the same results. Recovery or partial recovery of DNA synthesis occurred when the BPDE concentration was below 6 X 10( 4) mg/ml; at higher concentrations the cells were killed. Simultaneously replicating viral DNA's from viruses infecting the same cells before and after BPDE treatment of the cells exhibited the same reduced rate of synthesis. The evidence indicates that covalent adducts in viral DNA are not responsible for its reduced replication rate; moreover, it is probable that an insignificant number of adducts is produced in intracellular viral DNA at BPDE concentrations that do not kill CV-1 cells. Rather, it appears likely that BPDE inhibits viral DNA synthesis by attacking cellular DNA or non-DNA targets. Caution is therefore required in relating the effects of BPDE and other carcinogens to DNA adduct formation. PMID- 6297821 TI - Regioselective inhibition of benzo[a]pyrene metabolism by butylated hydroxyanisole. AB - The effects of butylated hydroxyanisole (BHA) on the metabolism of benzo[a]pyrene (BP) were investigated with mouse hepatic microsomes. Microsomes from BHA-fed mice showed a higher activity in catalyzing the formation of most metabolites from BP but a much lower activity in the formation of 9-hydroxy BP than control microsomes. Dietary BHA treatment enhanced the total metabolism of BP but decreased the microsomal metabolism of BP-7,8-diol, especially the formation of BP-trans-7,8-diol-anti-9,10-oxide. The altered metabolism of BP is believed to be due to the induction of new cytochrome P-450 species by dietary BHA. Consistent with this interpretation is the observation that the treatment also decreased the Km and increased the Vmax of ethoxycoumarin O-dealkylase. Short-term treatment (BHA administered p.o.) and in vitro added BHA were shown to inhibit BP metabolism. Thus, BHA can affect BP metabolism by exerting its inhibitory effect directly and by altering the composition of microsomal monooxygenase enzymes after a few days of exposure. PMID- 6297822 TI - Carcinogenic activity of benzo[a]pyrene and some of its synthetic derivatives by direct injection into the mouse fetus. AB - We have studied the carcinogenic effects of direct injection of benzo[a]pyrene (BP) and 6-methylbenzo[a]pyrene (BP6M) into fetal Swiss mice at 15 days of intrauterine growth. To determine the sensitivity of mouse fetuses to the action of ultimate carcinogens, benzo[a]pyrene-4,5-oxide (BPO) and a racemic mixture of 7 beta,8 alpha-dihydroxy-9 alpha, 10 alpha-epoxy-7,8,9, 10 tetrahydrobenzo[a]pyrene (BPDE), respectively, a proximal metabolite and an ultimate carcinogenic metabolite of BP, were also investigated in our system. The compounds were dissolved in acetone and trioctanoin (1:1) and injected at doses ranging from a minimum of 0.4 to a maximum of 19.8 nmol/fetus. Controls received 1 microliter/fetus of solvent. The experiment was terminated when the animals were 12-15 weeks old. On the basis of the percentages of lung adenomas and the average number of nodules/mouse, BPDE appears to be the most potent carcinogen of this group, causing 55% of tumors at a dose of 0.4 nmol and 73% at a dose of 4.0 nmol/fetus. At the dose of 0.4 nmol/fetus, neither BP nor BP6M induced tumors; injection of 4.0 or 19.8 nmol/fetus, caused incidences of 53% and 92%, respectively, in the case of BP6M and 24% and 39%, respectively, in the case of BP. BPO was not tumorigenic in this system, even at doses as high as 15.7 nmol/fetus. Eleven percent of the control animals had lung adenomas. The average number of nodules/mouse varied with the compound and the dose injected and closely followed the pattern of tumor incidence. PMID- 6297823 TI - Enhancement of replication of Epstein-Barr virus DNA in human lymphoblastoid cell lines by N-methyl-N'-nitro-N-nitrosoguanidine. AB - N-Methyl-N'-nitro-N-nitrosoguanidine (MNNG) is a potent carcinogen that alkylates nucleic acids. Interaction of MNNG with human lymphoblastoid cell lines carrying Epstein-Barr virus (EBV) was studied. Treatment of virus-producing cells (P3HR-1) with MNNG resulted in an approximately 3-fold increase in EBV genome copies per cell as determined by cRNA--DNA hybridization. This effect was not observed in a non-virus-producer line (Raji). Dose-response studies indicated that the optimum concentration was between 0.5 micrograms and 2 micrograms/ml. This same dose range was most effective in inhibiting cell proliferation both of P3HR-1 and Raji cells. Concomitant treatment of P3HR-1 cells with MNNG and 12-O tetradecanoylphorbol-13-acetate gave an additive increase to 9-fold of the number of EBV genome copies per cell. Pretreatment of Raji cells with MNNG followed by superinfection with P3HR-1 virus resulted in a 35% enhancement of EBV DNA replication as analyzed by density centrifugation. In contrast, Raji cells superinfected with MNNG-treated EBV showed a marked reduction in EBV DNA replication which indicates that the lesions produced in the viral genome by the drug interferred with the infectious potential of the virus. PMID- 6297824 TI - Preferential binding of polycyclic hydrocarbons to matrix-bound DNA in rat-liver nuclei. AB - The reactions of benzo[a]pyrene and 7,12-dimethylbenz[a]anthracene metabolites and of r-7,t-8-dihydroxy-t-9,10-oxy-7,8,9,10-tetrahydrobenzo[a]pyrene with the DNA of matrix-bound and released chromatin fractions of rat-liver nuclei have been examined. Qualitatively there were no differences between the DNA-bound metabolites in each fraction but more binding to matrix-bound DNA occurred. Evidence was obtained that the increased binding of hydrocarbon to matrix-bound DNA was not dependent upon the proximity of hydrocarbon-metabolizing enzymes and Sephadex LH20 chromatography showed that the differences between the fractions were not due to contamination of DNA with residual proteins. The conformation of the matrix-bound chromatin may make its DNA more accessible to reactive metabolites than that of released chromatin. PMID- 6297825 TI - Dosimetry of PAH skin carcinogenesis: covalent binding of benzo[a]pyrene to mouse epidermal DNA. AB - Benzo[a]pyrene (BaP) is metabolized to the chemically reactive anti and syn isomers of the 7,8-diol-9,10-epoxides of BaP (BPDE) which bind covalently to DNA to form DNA/BPDE complexes. Tetrols liberated from the DNA/BPDE complex by acid hydrolysis are easily quantified by h.p.l.c. using fluorescence detection. This approach allows femtomole amounts of BPDE associated with the DNA isolated from a single mouse to be detected using conventional instrumentation. The usefulness of this technique to estimate the interaction of BaP with DNA of mouse skin, both in the intact animal and in organ culture, was investigated. With mouse skin in organ culture it could be demonstrated that: (i) upon a single topical application of 5 micrograms of BaP, binding to DNA occurred via BPDE at a linear rate for up to 65 h, (ii) the amount of binding was dose dependent at concentrations of BaP less than 10 micrograms. PMID- 6297826 TI - A note on synthesis and reactivity towards DNA of glycidonitrile, the epoxide of acrylonitrile. AB - Different ways have been used to synthesize glycidonitrile, the epoxide of acrylonitrile. The reaction product has been characterized by n.m.r. spectroscopy. While glycidonitrile-induced strand breaks in SV40 phage DNA in vitro, acrylonitrile did not. PMID- 6297827 TI - Comparative studies of atrial and ventricular myosin from normal, thyrotoxic, and thyroidectomized rabbits. AB - Changes in enzymatic and structural properties of ventricular myosin in thyrotoxic rabbit hearts have been investigated extensively. However, there is little information regarding the effect of thyroid hormone on the atrial myosin. In this study, we have compared enzymatic and structural changes of ventricular and atrial myosin from euthyroid, hypothyroid, and hyperthyroid rabbits. In euthyroid rabbits, Ca++- and actin-activated ATPase activities of atrial myosin were 2-fold greater than those of the ventricular myosin. The Ca++- and actin activated ATPase activities of atrial myosin from hypothyroid and hyperthyroid rabbits were identical with the values for atrial myosin from euthyroid rabbits. The same ATPase activities of ventricular myosin decreased in hypothyroid hearts but increased in hyperthyroid rabbits. The K+ (EDTA)-ATPase activities of all myosins were the same, irrespective of the thyroid status of the animal. Pyrophosphate-polyacrylamide gel electrophoresis patterns showed two isoenzymes (designated as A1 and A3) of atrial myosin in euthyroid hearts. The same electrophoretic patterns also showed in atrial myosin from hypothyroid and hyperthyroid hearts. The ventricular myosin from euthyroid hearts also exhibited two isoenzymes (designated as V1 and V3) but each with slower electrophoretic mobilities than the corresponding atrial myosin. In hypothyroid hearts, only V3 isoenzyme was seen, whereas, in hyperthyroid hearts, only V1 isoenzyme was seen. These results suggest that thyroid hormone controls ventricular myosin ATPase activity by controlling synthesis of a specific ventricular isoenzyme, whereas thyroid hormone does not affect atrial myosin ATPase, possibly due to its inability to control atrial myosin synthesis. PMID- 6297828 TI - Mechanism of Ca++ antagonist-induced vasodilation. Intracellular actions. AB - We studied the effects of Ca++ antagonists on intact and skinned muscles of rabbit mesenteric artery. Intact muscle contractions were inhibited by 10(-6) M diltiazem, whereas greater levels were required to abolish contractions in skinned muscle fibers. In contrast, nisoldipine had no effect on skinned muscle contractions, although it inhibited, almost completely, the contraction of intact muscle at concentrations below 10(-6) M. In the presence of EGTA, norepinephrine induced contractions result from a release of Ca++ from an intracellular store. Diltiazem inhibited these contractions at concentrations between 10(-6) and 10( 4) M. Higher doses were required in studies with skinned muscle preparations. Unlike diltiazem, nisoldipine only partially inhibited the Ca++-free norepinephrine-induced contractions in the range of 10(-7) to 10(-5) M. From these results, we assumed that at low concentrations (below 10(-6) M), diltiazem induced relaxation by blocking Ca++ influx, whereas at relatively high concentrations (above 10(-6) M), an inhibition of Ca++ release from an intracellular store also occurred. A similar conclusion was reached regarding the mechanism whereby nisoldipine inhibits force developments. PMID- 6297830 TI - Renal and adrenal responses to hypoxemia during angiotensin-converting enzyme inhibition in lambs. PMID- 6297829 TI - Mechanism of adenosine inhibition of catecholamine-induced responses in heart. AB - The properties of adenosine inhibition of catecholamine-induced responses were investigated, using an isolated rat heart preparation. Perfusion of hearts with 0.1 microM isoproterenol increased myocardial cAMP content 2.8-fold, activation of cAMP-dependent protein kinase 4.4-fold, phosphorylase a formation 3.4-fold, left ventricular pressure 1.8-fold, rate of ventricular pressure development 2.1 fold, and rate of ventricular relaxation 2.2-fold within 1 minute. When perfused with the isoproterenol, 10 microM adenosine reduced the catecholamine-produced increase in cAMP, cAMP-dependent protein kinase, and phosphorylase by 30-40%, and the elevation in left ventricular pressure and rate of ventricular pressure development by 40-70% within 40 seconds. More than 2 minutes were required for the nucleoside to significantly reduce the isoproterenol-elicited increase in the rate of ventricular relaxation. Perfusion of adenosine alone at concentrations from 0.1 to 10 microM were without effect on the above parameters. Theophylline at 50 microM had no effect alone on the above parameters but blocked the inhibitory actions of adenosine on the isoproterenol-induced responses. In the presence of 15 mM Mg++ adenosine reduced by approximately 56% the 2-fold increase in myocardial membrane adenylate cyclase activity produced by 1 microM isoproterenol without affecting basal or fluoride-stimulated activity. Adenosine also reduced the isoproterenol-induced increase in enzyme activity assayed at 1-2 mM Mg++, a level that more closely approximates the intracellular activity of the ion. The results suggest that physiological concentrations of adenosine attenuate the catecholamine-induced increase in cAMP content, cAMP-dependent protein kinase activation, phosphorylase a formation, and contractile parameters in the working heart, via reducing the beta-adrenergic activation of adenylate cyclase. PMID- 6297831 TI - Interaction of verapamil and other calcium channel blockers with alpha 1- and alpha 2-adrenergic receptors. AB - To determine the specificity of the previously demonstrated competition of verapamil with radioligand binding to alpha-adrenergic receptors, we examined the interaction of calcium channel blockers with alpha 1- and alpha 2-adrenergic receptors on several tissues. Verapamil competed for [3H] prazosin binding to alpha 1-adrenergic receptors and for [3H]yohimbine binding to alpha 2-adrenergic receptors in several tissues (human platelets, rat kidney and heart, and cultured muscle cells) with dissociation constants of 0.6-6 microM. The calcium channel blockers D600, D591, fendiline, and prenylamine--which are structural analogues of verapamil--also competed for [3H]yohimbine binding to human platelets. Two other calcium channel blockers, diltiazem and nifedipine, did not compete for [3H] yohimbine binding to human platelets or [3H]prazosin binding to membranes prepared from rat ventricles. We used [3H]nitrendipine binding to identify putative calcium channels on rat myocardial membranes. Nifedipine and verapamil blocked these [3H]nitrendipine-binding sites on ventricular membranes, but epinephrine and prazosin did not, indicating that the ventricular alpha 1 receptors and calcium channels are distinct. We found no specific [3H]nitrendipine binding to human platelets. We conclude that the interaction of verapamil with alpha-adrenergic receptors is not receptor subtype or tissue specific, that interaction with alpha-adrenergic receptors is not a property of all calcium channel blockers, and that the interaction of verapamil with alpha adrenergic receptors and its interaction with calcium channels occur at at least two distinct sites. PMID- 6297832 TI - Agonist-induced endothelium-dependent relaxation in rat thoracic aorta may be mediated through cGMP. AB - The present study investigates the hypothesis that endothelium-dependent relaxation of vascular smooth muscle may be mediated through the formation of cGMP. Relaxation of the rat thoracic aorta to acetylcholine, histamine, and Ca++ ionophore A23187 was associated with increased levels of cGMP in a time- and concentration-dependent manner, whereas cAMP levels were unaltered. Removal of the endothelium prevented relaxation to these agents and prevented the increased levels of cGMP. Removal of the endothelium after exposure to acetylcholine only partially decreased the elevated levels of cGMP, suggesting that the changes in cGMP occurred within the smooth muscle cells. Eicosatetraynoic acid, an inhibitor of lipoxygenase and cyclooxygenase, and quinacrine, an inhibitor of phospholipase, prevented and reversed acetylcholine-induced relaxation, respectively, and inhibited acetylcholine-induced increased levels of cGMP. In contrast, sodium nitroprusside-induced relaxation and increased levels of cGMP were independent of the presence of the endothelium, exposure to eicosatetraynoic acid, and quinacrine. The present results support the hypothesis that vascular smooth muscle relaxation induced by some agents is dependent on the presence of the endothelium and is mediated through the formation of an endothelial factor that increases cGMP levels in smooth muscle. PMID- 6297833 TI - Further evaluation of luminol-enhanced luminescence in the diagnosis of disorders of leukocyte oxidative metabolism: role of myeloperoxidase. AB - Chemiluminescence can be used to identify defects in the oxidative metabolism of granulocytes. This procedure has recently been adopted for use with microliter quantities of whole blood, appropriate for prenatal or neonatal study. Although the contribution of myeloperoxidase to the chemiluminescence assay has been noted, the possible diagnostic confusion between chronic granulomatous disease of childhood (which is rare and severe) and myeloperoxidase deficiency (which is common and of little clinical consequence) has not been stressed. We report a father and his infant daughter whose cells emitted no light in the luminol enhanced luminescence assay; both patients are totally peroxidase deficient. These results emphasize the hereditary nature of myeloperoxidase deficiency, and the possibility for erroneous diagnosis of chronic granulomatous disease of childhood based on the luminol-enhanced luminescence test. PMID- 6297834 TI - An evaluation of four serum tests for pregnancy. AB - We evaluated four pregnancy tests (Biocept-G, Beta-CG, Preg/Stat, and HCG-Beta Screen), using sera from 59 nonpregnant subjects and 77 patients with serum human choriogonadotropin beta-subunit (beta-hCG) concentrations ranging from 4 to 100 000 int. units/L. The results obtained for each test were compared with the results predicted on the basis of the sample's beta-hCG concentration and the beta-hCG concentration the manufacturer claimed necessary for a positive result (the test's analytical sensitivity). Biocept-G had the best sensitivity (100%), specificity (98.9%), and accuracy (99.2%). Beta-CG had the poorest sensitivity (86.4%), Preg/Stat the poorest specificity (87.5%), and accuracy (92.6%). We confirmed the manufacturer's claimed analytical sensitivity (200 int. units/L) for the Biocept-G procedure, but our calculated analytical sensitivity for the other tests was significantly different from that claimed by their manufacturers. Best results were obtained with Biocept-G, but with its analytical sensitivity of 200 int. units/L, samples from early pregnancy will give negative results. None of the pregnancy tests evaluated here will establish the presence or absence of early pregnancy with certainty. PMID- 6297835 TI - Separation of the high-molecular-mass "immunoreactive ACTH" measured with the CIS Sorin Kit by ultracentrifugation of plasma. PMID- 6297836 TI - [Enzyme immunoassay of beta-endorphin]. PMID- 6297837 TI - [Ellsworth-Howard test by using 1-34h PTH as parathyroid hormone during childhood]. PMID- 6297838 TI - Characteristics of human soft tissue sarcomas in xenografts and in vitro. AB - Two permanent, transplantable strains of rhabdomyosarcoma (SAS) and synovial sarcoma (KUSHI) were established by transplanting them serially into nude mice. The original SAS tumor and its transplant were histologically identical and gave an appearance of so-called embryonal rhabdomyosarcoma. The KUSHI tumor changed after the second passage, showing the biphasic pattern of synovial sarcoma, while undifferentiated myoblast-like cells were observed throughout the original tumor. Three in vitro cell lines from the SAS and one from the KUSHI tumor were successfully established. The human origin of the SAS and KUSHI transplanted tumors could be verified by the presence of human chromosomes. The cells cultured from the SAS tumor were more sensitive to anticancer drugs than those of the KUSHI tumor. However, as the number of serial passages on nude mice increased, the sensitivities of the SAS cells decreased, while those of the KUSHI cells increased. The modal chromosome number of the SAS tumor changed from 51 to 49 to 46 during serial passages on nude mice. These observations suggest that the sensitivities to anticancer drugs not only differ among soft tissue sarcomas of different histologic types, but also change continuously during the growth of the tumor itself. PMID- 6297839 TI - [Two cases of Epstein-Barr virus encephalitis with action tremor in elderly patients]. PMID- 6297840 TI - Renal allograft recovery subsequent to apparent "hyperacute" rejection based on clinical, scintigraphic, and pathologic criteria. AB - An unusual case is described in which in spite of clinical, scintigraphic and histologic findings strongly supportive of a diagnosis of "hyperacute rejection," recovery of renal function occurred. These findings are in contrast to the current literature in which it is generally accepted that a renal allograft showing neither pertechnetate transit nor hippurate concentration warrants allograft nephrectomy irrespective of the etiology. Scintigraphic evaluation included both dynamic studies after a bolus administration of Tc-99m pertechnetate and serial renogram collections after the intravenous administration of I-131 Hippuran. PMID- 6297841 TI - Effects of charcoal, sodium bicarbonate, and ammonium chloride on chlorpropamide kinetics. AB - The effects of activated charcoal, sodium bicarbonate, and ammonium chloride by mouth on chlorpropamide kinetics was studied in six healthy subjects. Activated charcoal, 50 gm, given immediately after 250 mg chlorpropamide reduced its absorption by 90%, but when given in repeated doses from 6 hr on (50 gm followed by 12.5 gm at 6-hr intervals) it did not shorten the chlorpropamide half-life (t 1/2). The t 1/2 of chlorpropamide was shortened from 49.7 +/- 7.4 to 12.8 +/- 1.1 hr by sodium bicarbonate and prolonged to 68.5 +/- 10.5 hr by ammonium chloride. The 72-hr urinary excretion of chlorpropamide was increased fourfold by alkalinization and decreased to 1/20 of baseline by acidification of the urine. The renal clearance of chlorpropamide correlated with urinary pH, ranging from 1 to 1000 ml/hr at the pH from 5 to 8. Urinary pH is likely to explain at least a part of great interindividual differences in the serum chlorpropamide concentrations during steady-state and variations in amount of urinary metabolites. In chlorpropamide intoxications activated charcoal can reduce absorption and alkalinization of the urine accelerates its elimination. PMID- 6297842 TI - Abrasivity of sodium bicarbonate. PMID- 6297843 TI - Gallium scanning in the diagnosis of hepatocellular carcinoma: a clinicopathological study of 45 patients. AB - Gallium-67 citrate scintigraphy was carried out in 45 patients with primary hepatocellular carcinoma in conjunction with serum alpha-fetoprotein measurement, 99mTc-colloid liver scan and selective arteriography. 67Ga accumulated in the tumour in 31 patients, and partial uptake was noted in three others. Scanning detected small hepatomas in four patients with cirrhosis in whom 99mTc-colloid scanning was negative. Clinical, laboratory and histopathological comparisons of 67Ga-positive and -negative patients showed no significant differences. 67Ga scanning of the liver was found to be useful in the diagnosis of hepatocellular carcinoma, especially in patients with associated cirrhosis. PMID- 6297844 TI - A micro-method for the study of acid secretory function in isolated human oxyntic glands from gastroscopic biopsies. PMID- 6297845 TI - Immunogenicity of foot-and-mouth disease virus type O1 replicated in either monolayer or suspended BHK cell system. AB - The efficacy of vaccines formulated from the 10th passage of foot-and-mouth disease virus (FMDV) type O1 in monolayer baby hamster kidney (BHK) cells and the 8th passage in suspension BHK cells was compared in steers. The vaccines were inactivated with ethylenimine, contained an equal amount of antigen and were emulsified in oil-adjuvant. Six animals were vaccinated with each vaccine. During the challenge of immunity (91 days post-vaccination, DPV), one out of the six steers from the monolayer vaccine group became infected with the challenge virus while none of the six steers from the suspension vaccine group contracted the disease during the test period. The neutralizing antibody titers (means) of the serum samples taken at different DPV also did not suggest significant variation between these vaccines. In addition exposure to FMDV infected animals demonstrated that both vaccines elicited an immune state in the vaccinates. PMID- 6297846 TI - [Enzootic bovine leukosis application of E.L.I.S.A. technique for the detection of antibodies]. AB - A simple microplate method of enzyme-linked immunosorbent assay (E.L.I.S.A.) for detecting antibodies to bovine leukosis virus (B.L.V.) is described. The antigen consisted of a solution containing the two major antigens of the B.L.V. (gp 51 and p 24) obtained by a technique of purification using CN-Br activated Sepharose 4B. This E.L.I.S.A. was compared with the agar gel immunodiffusion test (A.G.I.D.T.) in a study of 545 bovine sera. The total discrepancy rate between the two tests was 11% with a better sensitivity for E.L.I.S.A. PMID- 6297847 TI - Enzyme-linked immunoassay in the diagnosis of leptospirosis in domestic animals using peroxidase-conjugated protein-A. AB - The ELISA test for detection of antibodies to Leptospirosis in domestic animals was performed using Staphylococcal protein-A coupled to peroxidase in place of antisera to IgG. Genus- and type-specific antigens were extracted with SDS technique from four pathogenic serotypes and two non-pathogenic ones, and they were identified with the aid of ELISA using specific rabbit antisera. Micro agglutination (MA) and ELISA were compared using a total of 48 positive swine sera and a 100% agreement was obtained, since with sera from 16 dogs clinically suspected of Leptospirosis the ELISA resulted highly more sensitive and precocious than MA in detecting specific antibodies. PMID- 6297848 TI - LOCALIZE: computer-assisted localization of peripheral nervous system lesions. PMID- 6297849 TI - Rates of synthesis of basement membrane proteins by differentiating teratocarcinoma stem cells and their modulation by hormones. AB - The embryonal carcinoma mouse cell line F-9 was used as a convenient model for a quantitative study of the production of the basement membrane proteins laminin and type IV collagen. Both proteins could be identified in the culture medium and cell layer by radioimmuno assays, metabolic labeling and immunofluorescence. More than 95% of the material is secreted into the medium. Lack of ascorbic acid inhibits secretion of type IV collagen but not of laminin. Induction of differentiation into endoderm-like cells by retinoic acid consistently caused after a lag period of 2-3 days a 5-10 fold increase in the production of basement membrane proteins but not of total protein. Dibutyryl cyclic AMP further potentiated this specific effect particularly with respect to type IV collagen synthesis. Insulin, epidermal growth factor and nerve growth factor produced only moderate increases (10-60%) in the amount of laminin and type IV collagen. Effects of these hormones were only observed with certain doses and were quite variable between different experiments. PMID- 6297850 TI - Partial characterization of collagenous and noncollagenous basement membrane proteins synthesized by the 14.5-day rat embryo parietal yolk sac in vitro. AB - Parietal yolk sacs isolated from 14.5-day rat embryos and incubated in vitro with either [14C]proline, [3H]mannose or 3H-labeled amino acid mixture synthesized and secreted basement membrane collagenous and noncollagenous glycoprotein components with relative molecular weights of 350,000 (350K), 220,000 (220K), 185,000 (185K), 175,000 (175K) and 150,000 (150K). The 185K and 175K components appeared to be similar to the pro-alpha 1 (IV) and pro-alpha 2(IV) chains, respectively, which have been isolated from other sources. These components were completely susceptible to bacterial collagenase, but were only partially susceptible to alpha-chymotrypsin digestion. The 350K and 220K components appeared to be similar to subunits of laminin (or PYS A and PYS B, respectively) which have been characterized by others, while the 150K component may be similar to entactin (or PYS C). These components were completely resistant to bacterial collagenase and completely susceptible to alpha-chymotrypsin digestion. In addition, the basement membrane of the parietal yolk sac (Reichert's membrane) stained intensely with antibodies directed against either rat laminin or mouse basement membrane procollagen. The results of these experiments suggest that the 14.5-day rat embryo parietal yolk sac is a useful system for studying the structure, biosynthesis and deposition of basement membrane components. PMID- 6297852 TI - Modelling the control mechanism of the sodium channel in the apical membrane of tight epithelia. PMID- 6297853 TI - Conserved linkage of soluble aconitase and galactose-1-phosphate uridyl transferase in mouse and man: assignment of these genes to mouse chromosome 4. AB - Polymorphism and linkage of mouse soluble aconitase (Aco-1) and galactose-1 phosphate uridyl transferase (Galt) are reported. Three alleles at each locus were recognized on the basis of differences in electrophoretic mobility. Linkage crosses involving Aco-1, brown (b), major urinary protein (Mup-1), and the Rb(4.6)2Bnr Robertsonian translocation (Rb2) showed that Aco-1 is located on the proximal portion of chromosome 4 between (Mup-1) and the centromere. Because Aco 1 and Galt are located on the short arm of the human 9, linkage of mouse Aco-1 and Galt was thought to be likely. Linkage crosses involving Rb2, Galt, Aco-1, Mup-1, and b confirmed this expectation and established the order of loci as: centromere--Galt--Aco-1--Mup-1--b. The conserved linkage of Aco-1 and Galt in mouse and man probably represents an ancient linkage not yet disrupted by chromosomal rearrangement, rather than a linkage preserved by natural selection. PMID- 6297851 TI - Calcium, the heart, and calcium antagonists. PMID- 6297854 TI - Assignment of the human phosphoserine phosphatase gene (PSP) to the pter leads to q22 region of chromosome 7. AB - Phosphoserine phosphatase (PSP) catalyzes the hydrolysis of phosphoserine to serine. PSP expression has been examined in human-mouse somatic cell hybrids retaining different combination of human chromosomes. Human PSP is expressed only when the pter leads to q22 segment of the human 7 and its enzyme marker beta glucuronidase (GUSB) are retained in cell hybrids. The structural gene, PSP, is therefore assigned to this region of the human 7. PMID- 6297855 TI - The human phosphoserine phosphatase gene (PSP) is mapped to chromosome 7 by somatic cell genetic analysis. AB - Human phosphoserine phosphatase (E.C. 3.1.3.3) (PSP), a phosphohydrolase, participates in the biosynthesis of serine from carbohydrates (Moro-Furlani et al., 1980). Starch gel electrophoresis has identified isoenzymes and rare allelic variants. More common electrophoretic variation is thought to be due to secondary nongenetic modification. Using starch gel electrophoresis of interspecific somatic cell hybrids, we have been able to assign the gene for this enzyme to human chromosome 7. PMID- 6297856 TI - Mapping of the human gene for epidermal growth factor receptor (EGFR) on the p13 leads to q22 region of chromosome 7. AB - We previously reported that the structural gene for epidermal growth factor receptor (EGFR) can be mapped to the p22 leads to qter region of human chromosome 7 (Shimizu et al., 1979, 1980). In the present study, we produced two series of human-mouse cell hybrids by fusing mouse A9 cells that are deficient in EGFR with the human diploid fibroblast lines GM1356, 46,XX,t(1;7)(p34;p13), and GM2068, 46,XX,t(6;7)(q27;q22), both of which possess EGF receptors. Expression of EGF binding ability in the former series of cell hybrids was correlated with the retention of the human translocation chromosome containing the 7p13 leads to qter region, and in the latter series of cell hybrid it was correlated with the retention of the human translocation chromosome containing the 7pter leads to q22 region. Therefore, the EGFR gene can be localized in the p13 leads to q22 region of chromosome 7. PMID- 6297857 TI - Radionuclide imaging in myocardial sarcoidosis. Demonstration of myocardial uptake of technetium pyrophosphate99m and gallium. AB - A patient had severe congestive cardiomyopathy secondary to myocardial sarcoidosis. The clinical diagnosis was confirmed by radionuclide ventriculography, thallium 201 (201Tl), gallium-67 (67Ga), and technetium-99m (99mTc) pyrophosphate (TcPYP) scintigraphy. Myocardial TcPYP uptake has not been reported previously in sarcoidosis. In this patient, TcPYP was as useful as gallium scanning and thallium imaging in documenting the myocardial process. PMID- 6297858 TI - Combination therapy of experimental candidiasis, cryptococcosis and aspergillosis in mice. AB - Combination pairs of the major systematic antimycotic drugs, amphotericin B (AmphB), 5-fluorocytosine (5-FC) and ketoconazole (Ktz) were administered to mice with experimental candidiasis, cryptococcosis and aspergillosis at a variety of combination ratios. The 3 mycoses were produced with 3 strains each of Candida albicans, Cryptococcus neoformans, and Aspergillus fumigatus, respectively, which were preselected to represent 3 different degrees of 5-FC sensitivity ('normally sensitive', 'moderately resistant', and 'definitely resistant'). The life prolonging effect of the combinations was compared with the effect of each partner administered alone at the same and at the double dosage. Using the U test of Mann and Whitney and setting limits which on the whole were more rigorous than those of the isobole methods commonly applied to the study of drug interactions, the effects of the concentrations were classified as 'synergistic', 'additive', 'indifferent' or 'antagonistic'. The combination AmphB plus 5-FC was definitely synergistic or definitely additive in all 3 candidiasis models, the most pronounced synergism occurring in the infection with the 'definitely 5-FC resistant' C. albicans strain; in cryptococcosis produced by any of the 3 C. neoformans strains the effect was definitely additive, but only slightly additive or indifferent in the 3 aspergillosis models. The combination AmphB plus Ktz was slightly synergistic in candidiasis produced by one C. albicans strain, but definitely antagonistic in this mycosis produced by the remaining 2 strains of the same species; the combination was definitely additive or, even, slightly synergistic in the 3 cryptococcus models, but, again, antagonistic in aspergillosis produced by all 3 strains of A. fumigatus. 5-FC plus Ktz was additive or indifferent in the 3 candidiasis models, but throughout indifferent in cryptococcosis and aspergillosis. PMID- 6297859 TI - Clinical presentation of candidal balanitis--its differential diagnosis and treatment. AB - The clinical presentation of candidal balanitis is discussed. Differential diagnosis from other forms of balanitis and dermatoses affecting the genitals is made. Treatment of candidal balanitis is then mentioned. General measures, contact investigation and specific measures such as the polyene antibiotics and the newer imidazole group of antifungal drugs are exemplified. PMID- 6297860 TI - [Industrial and pharmaceutical novelties. Tres-Pres toothpaste]. PMID- 6297861 TI - Characterization of X-chromosome specific satellite DNA of Muntiacus muntjak vaginalis. AB - A highly repeated DNA (designated satellite IA) was isolated from cultured cells of Muntiacus muntjak vaginalis and its organization analyzed by the use of restriction nucleases and hybridization experiments with cloned DNA-fragments. Several restriction nucleases cleave the satellite IA DNA into a series of fragments, which are multiples of a basic repeat unit of 800 bp. Sequences homologous to the satellite IA DNA were also found in a second highly repetitive DNA component of Muntiacus muntjak vaginalis (satellite IB). Its organization is more complex than the one of satellite IA and does not conform to a simple periodicity of a basic repeat unit.--Hybridization in situ revealed, that both satellites are confined in their entirety to the X-chromosome, where they are located at both arms close to the centromere. No satellite DNA was found at the Y1-chromosome, which is considered to be homologous to the long arm of the X chromosome. These results have interesting implications for the evolution of the X-chromosome. PMID- 6297862 TI - Hypocalcemia and steatorrhea--clues to etiology. AB - Two boys with idiopathic hypoparathyroidism had extensive studies of gastrointestinal function during hypocalcemia accompanied by steatorrhea. No evidence of generalized gastrointestinal moniliasis or abnormal mucosal structure or function was observed. Studies of pancreatic function and bile salt metabolism during hypocalcemia demonstrated deficient meal-stimulated intraluminal pancreatic enzyme concentrations in both subjects and reduced bile salt concentrations in one subject. However, following stimulation with exogenous octapeptide of cholecystokinin, intraluminal pancreatic enzyme and bile salt concentrations were normal in both. Cholic acid pool sizes were markedly increased in both subjects during hypocalcemia (9 and 12 times larger than during normocalcemia) and cholic acid turnover was reduced during hypocalcemia in one subject. Our findings suggest that during hypocalcemia, insufficient endogenous cholecystokinin is released by the duodenal mucosa during a meal stimulus to stimulate normal gallbladder contraction and pancreatic enzyme secretion. PMID- 6297864 TI - [4 rare localizations of mixed tumors in the ORL area]. PMID- 6297865 TI - [Focus of human trypanosomiasis in Vavoua, Ivory Coast. Recent data on the epidemiology of the disease in relation to the ecology of Glossina]. PMID- 6297863 TI - Vitamin D deficiency, osteomalacia, and primary biliary cirrhosis. Response to orally administered vitamin D3. AB - Five patients with primary biliary cirrhosis and vitamin D deficiency (serum 25 hydroxyvitamin D less than 6 ng/ml) are presented. All patients had low serum 24,25-dihydroxyvitamin D3 concentrations. Three patients had histological osteomalacia, negative calcium balance, and subnormal serum 1,25-dihydroxyvitamin D3. Malabsorption of a standard dose of [3H]vitamin D3 was found in three of four patients with steatorrhea, enabling the effective dose of vitamin D3 given to be calculated. Oral vitamin D3 400-4000 IU/day (effectively 400-1860 IU/day) resulted in a return to normal of the serum vitamin D metabolites, correction of the impaired intestinal calcium absorption and healing of the osteomalacia. Increases in serum calcium, phosphate, and the renal tubular reabsorption of phosphate occurred with a concomitant decrease in serum parathyroid hormone. It is suggested that osteomalacia in primary biliary cirrhosis is the end result of vitamin D deficiency; the hepatic and renal hydroxylations of vitamin D are normal and target tissues are responsive to endogenously produced metabolites of vitamin D. PMID- 6297866 TI - The possible chelating effect of sodium diethyldithiocarbamate (DDC) in nickel allergic patients. Patch test procedures with nickel on DDC pretreated skin. AB - Encouraged by some reports about the chelating effect of p.o. given sodium diethyldithiocarbamate (DDC) in nickel allergic patients, some experiments were performed to determinate the blocking effect of locally applied DDC on positive patch test reactions with nickel. Two groups of nickel allergic patients were patch tested with different concentrations of nickel sulphate on with DDC treated skin. Control patch tests were done on areas which were pretreated with the creambase only. In one group 10% DDC in the cream was applied under 24 h occlusion; in this group also a nickel coin was tested. In the second group of patients the DDC cream was rubbed four times during 24 h without occlusion. In both groups there were no statistical differences regarding the reactions on skin pretreated with DDC and with the cream base. From this investigation it may be concluded that there is no indication that a DDC-containing barriercream will be of great help in patients with occupational dermatitis, which is especially due to nickel sensitization. PMID- 6297867 TI - [Ranitidine, cimetidine and the prevention of stress ulcers]. PMID- 6297868 TI - [Tumor markers in C-cell cancer]. AB - C-cell carcinoma (medullary thyroid carcinoma) is associated with increased serum calcitonin levels. In addition, increased serum levels of other tumour markers are also found: carcinoembryonic antigen (CEA), adrenocorticotrophic hormone (ACTH) and tissue polypeptide antigen (TPA). The relevance of these tumour markers as compared to calcitonin was evaluated in 46 patients with C-cell carcinoma. With decreasing frequency pathologically increased levels were found: calcitonin 93%, CEA 73%, TPA 17% and ACTH 8%. There was a positive correlation between calcitonin and CEA values. Calcitonin and CEA levels correlated with the clinical tumour stage. In selective venous catheterisation estimation of calcitonin is superior to estimation of CEA. Assessment of CEA supplements calcitonin measurements in follow-up controls and evaluation of success of treatment. As regards specificity and sensitivity calcitonin is superior to all other tested tumour markers in C-cell carcinoma. PMID- 6297869 TI - [Causes of hypoglycemia in extrapancreatic tumors]. PMID- 6297870 TI - [The value of radiotherapy after modified radical mastectomy. 10-year results of the Heidelberg study]. AB - Modified radical mastectomy was performed in 142 female patients with operable carcinoma of the breast. One of the two prospectively randomised groups received postoperative radiotherapy with 6000 rad. Survival rates were equal for both groups. Follow-up radiotherapy had a beneficial influence on survival rate and rate of distant metastases in median tumour sites. In lateral tumour localization the course of disease was less favourable with radiotherapy than without. Local relapse rates were slightly, however not significantly, lower after irradiation. The decisive prognostic criterium is T- and N-stage at the time of diagnosis. PMID- 6297872 TI - Effects of maternal protein deficiency on synaptic plasma membranes in offspring. AB - This study examined the influence of maternal protein deficiency during lactation on the accretion and synthesis of synaptic plasma membrane (SPM) proteins in developing offspring. It also examined the activity of Mg2+-dependent, ouabain sensitive (Na+-K+)-ATPase in SPMs. The results of this study demonstrated that early postnatal undernutrition produces a significant deficit of SPM proteins. Despite this deficit, 10- to 20-day-old undernourished rats incorporated more [3H]-leucine into SPM proteins than age-matched control rats. The specific activity of (Na+-K+)-ATPase was decreased in SPMs from undernourished rats early in development, but was increased at 20 and 34 days of age. Interestingly, 20- and 34-day-old undernourished rats also had an increased proportion of [3H] leucine-derived radioactivity in the band on SDS gels which corresponded to (Na+ K+)-ATPase. In summary, it appears that maternal deficiency of protein during lactation affects the synaptic plasma membranes in offspring quantitatively and qualitatively. PMID- 6297873 TI - Differences between GABA receptor binding to membranes from cerebellum during postnatal development and from cultured cerebellar granule cells. AB - GABA receptor binding sites were determined in membranes from cerebella of 7-, 15 and 60-day-old rats and cerebellar granule cells derived from 7-day-old rats and cultured for 8 days using [3H]GABA as the ligand and nonradioactive GABA to measure nonspecific binding. Membranes from cerebellum at all postnatal stages exhibited at least two binding sites for GABA with binding constants around 7-9 and 150-750 nM, respectively. The total number of binding sites expressed on the basis of total protein increased 10 times between 7 and 60 days of age and 2 times between 15 and 60 days. The latter increase could be quantitatively accounted for by an increase in the number of low affinity sites. In contrast to this, membranes from cultured granule cells only exhibited the high affinity binding site (KD 7.1 +/- 0.5 nM) and the number of binding sites was comparable to that of cerebellar membranes from rats of the corresponding age (15 days old). GABA analogues such as muscimol, piperidine-4-sulphonic acid, homo-beta-proline, THIP, and isoguvacine were potent displacers of GABA binding to cultured granule cells, whereas nipecotic acid and guvacine had no effect. PMID- 6297871 TI - Antifungal agents used in systemic mycoses. Activity and therapeutic use. AB - The development of the polyene antibiotic, amphotericin B, provided for the first time a drug which was clinically effective in many serious mycotic diseases. Unfortunately, it requires parenteral administration and is often toxic, factors which limit the total cumulative dose which can be given. Efforts to utilise combinations of amphotericin B with other agents were best realised with amphotericin B/flucytosine in cryptococcal meningitis, and to a lesser degree in systemic candidiasis. More recently, the introduction of new imidazoles has extended the range of applications of these drugs to fungal diseases. Two members of this group, miconazole and ketoconazole, are promising agents. Miconazole is a parenterally administered agent for patients acutely ill with candidiasis and other mycotic infections. It may be the drug of choice for Petriellidium boydii infections and it is an attractive alternative to amphotericin B for intrathecal administration to patients with fungal meningitis. Ketoconazole offers much less toxicity, the advantage of oral administration, and the possibility of indefinitely prolonged therapy. However, it does not attain high concentrations in either the urine or cerebrospinal fluid. With the imidazoles, we have entered a new era of antifungal therapy which may produce even better antifungal agents than those currently available. PMID- 6297874 TI - Multiple excitability functions in cultured mouse spinal neurones. PMID- 6297875 TI - Neurophysiological studies of n-hexane polyneuropathy in the sandal factory. AB - Among 93 cases of n-hexane polyneuropathy during a large outbreak in 1968 44 cases were studied by conventional electromyogram and measurement of peripheral nerve conduction velocities, both of which showed remarkable changes, especially in the lower rather than the upper extremities. Over a few years since 1968 most of the cases completely recovered, except for a few with mild sensory impairment, after providing for 100 ppm as the maximal allowable concentration of n-hexane and well equipped ventilation systems in individual houses. During the rescreening in 1981, before which there occurred only 2 patients, 21 cases with mild n-hexane polyneuropathy were clinically and neurophysiologically observed, revealing mostly the same features as in the previous outbreak in 1968. From these data it may be suggested that, in spite of less than 50 ppm of n-hexane concentration in a room, the sandal workers have suffered from neurotoxicity from this organic solvent and the appropriateness of the present regulation on n hexane has to be reconsidered. PMID- 6297876 TI - Mechanisms for preserved nerve function with ischaemia in peripheral neuropathy. PMID- 6297877 TI - Peripheral nerve conduction and toxic effects on the motor unit. PMID- 6297878 TI - Genetic variation in testicular LH receptors in the mouse. AB - The concentration of testicular hCG (LH) binding sites was measured in five mutants with defective endocrine and/or reproductive functions, in a hypoandrogenic inbred strain and in six normal strains. A decrease in hCG binding was found in dw/dw mice, and an increase in Tfm/y and C57Bl10/J mice. Analysis of the results indicates the existence of a genetic factor(s) involved in the regulation of hCG (LH) binding. PMID- 6297879 TI - Dynamics of adrenal steroidogenesis in childhood: changes in the excretion of 16 oxygenated- and 11-oxygenated-steroids by 21-hydroxylase deficient children at various ages. AB - The close relationship between C19O2 steroid excretion and the ratio of C19O2/C21O5 steroids indicates that it is the elevation of C17,20-lyase activity, which represents the decisive factor in the cessation of androgen-corticoid disproportion during the puberty of healthy children. In 3-6 year-old C21 hydroxylase deficient children treated with corticoids in doses only partially suppressing endogenous ACTH secretion, the excretion of total C19 steroids increased continuously parallel with a well defined elevation of 16 alpha oxygenated-C21 steroid excretion. The patients did not show the physical signs of adrenarche before six years. This can be attributed to three factors: a) substitutional corticoid therapy; b) intraglandular control of delta 5-pathway by 16 alpha-hydroxylation of C21 steroids; c) neutralisation by 11 beta hydroxylation of the androgen effect of C19O2 steroids. PMID- 6297880 TI - Responses of salivary cortisol levels to stress-situations. AB - A procedure is described for determining salivary cortisol levels by a competitive protein-binding assay using horse transcortin. The collection of saliva was performed by means of filter paper-strips. Filter paper samples are more than 5 days stable after air-drying. In this form, the samples could be stored without refrigerator or deep-freezer and, if necessary, sent by post to the laboratory without any special precaution. Stressful situation of either painful or anxious origin were associated with an adequate increase of salivary cortisol levels. The increases were 157 to 230% of the initial or normal values dependent on the kind of stress. The mean values in 4 cases of Cushing's syndrome were 380% and 1 hour after 25 I.U. ACTH 690% higher than those in normal persons. In normal persons, a well-defined circadian rhythm has been observed. PMID- 6297881 TI - Electron-spin resonance study of mainstream and sidestream cigarette smoke: nature of the free radicals in gas-phase smoke and in cigarette tar. AB - Radicals in the gas phase of both mainstream and sidestream cigarette smoke have been studied using electron-spin resonance ESR spin-trapping techniques with alpha-phenyl-N-tert-butyl nitrone (PBN) as the spin trap. The principal radicals we trap appear to be alkoxyl radicals. Mainstream and sidestream gas-phase smoke each have about the same concentration of radicals, about 1 X 10(16) radicals per cigarette (or 5 X 10(14) per puff). These radicals are reactive, yet they appear to be remarkably long-lived: they are still spin trapped from gas-phase smoke after more than 5 min. We propose that a steady-state concentration of reactive radicals exists in gas phase cigarette smoke. We suggest that this steady state is produced by the slow oxidation of nitric oxide (present in high concentrations in smoke and relatively unreactive) to the more reactive nitrogen dioxide, followed by the reaction of nitrogen dioxide with reactive organic molecules in smoke (such as olefins and dienes). Preliminary experiments reported here support this hypothesis. Tar from both mainstream and sidestream smoke contains persistent free radicals that exhibit broad, single-line ESR spectra with g values of 2.003. The tar radical can be extracted into tert-butylbenzene and other organic solvents, and we have applied a variety of fractionation procedures to these solutions. Most of the radicals occur in the fractions that contain the phenolic tobacco leaf pigments. Treatment of alcoholic solutions of tar with base generates a new group of radicals that appear to be semiquinone radicals derived from the oxidation of the phenolic and polyphenolic species in tar. PMID- 6297882 TI - Research needs for hepatic injury due to environmental agents. AB - This report presents an overview of the patterns of liver injury produced by presently recognized toxic substances in the environment. Current concepts of the metabolic disposition of these substances in the liver suggest that their toxic effects may be modulated by an interplay between enzymatic pathways for activation or inactivation of foreign compounds. A better understanding of the genetic control of the enzymes involved in these pathways, as well as the mechanisms of cell death or malignancy produced by the products of these pathways, is needed. Although a number of sensitive biochemical tests of liver function are currently available, development of specific, noninvasive screening tests for the earliest manifestations of liver injury are urgently needed. PMID- 6297883 TI - Lung function and nervous system involvement in asbestosis. AB - A group of 115 patients with asbestosis of the lungs was tested in detail for neurological functions and lung functions in order that any possible impaired pulmonary function and neurological signs (especially peripheral neuropathy) could be discovered. Attention was also paid to finding out whether peripheral neuropathy, which is frequently encountered among asbestosis patients, had any role in decreased pulmonary function. A slight trend towards lowered spirometrical values (vital capacity, forced expiratory volume in one second, maximal mid-expiratory flow and maximal expiratory flow at 25% of vital capacity) was found for the group of patients with peripheral neuropathy. No differences in diffusing capacity were found between the groups of patients either with or without peripheral neuropathy. We concluded that the spirometrical findings are more indicative of a parallel phenomenon than a causal factor. Nor was the involvement of the central nervous system associated with decreased pulmonary function. In short, the pulmonary dysfunction of our asbestosis patients did not appreciably contribute to the occurrence of nervous system involvement, whether central or peripheral. PMID- 6297884 TI - Binding of fibronectin to the surface of group A, C, and G streptococci isolated from human infections. AB - Sixty-nine haemolytic and non-haemolytic streptococcal strains were isolated from various human infections and serogrouped with the coagglutination test. The amount of 125I-fibronectin bound to bacterial cells in a standard assay was determined. Most of the group A, C, and G strains were able to bind fibronectin. None of the group B or D strains bound significant amounts of fibronectin. Group A, C, and G streptococci appear to preferentially bind the N-terminal region of the fibronectin molecule because the 25K N-terminal fragment of the protein could effectively inhibit the binding of 125I-fibronectin to cells. Furthermore, the ability of representative strains of group A, C, and G to bind fibronectin was markedly reduced after trypsin treatment of the cells. Fibronectin binding components released from one strain by trypsin treatment inhibited the binding of 125I-fibronectin to all group A, C, and G streptococci strains. These findings indicate similarities among fibronectin binding proteins of the three groups of streptococci tested. However, the relative susceptibility to trypsin of fibronectin receptors of the three strains differed as did the relative potency of the inhibitory activity of receptors solubilized from different strains. Binding of fibronectin to the cell surface of group A, C, and G streptococci may contribute to virulence, for instance by promoting specific attachment to exposed fibronectin in open wounds and tissue lesions. PMID- 6297885 TI - A kinetic study of pyrophosphate: fructose-6-phosphate phosphotransferase from potato tubers. Application to a microassay of fructose 2,6-bisphosphate. AB - Pyrophosphate : fructose-6-phosphate phosphotransferase (PPi-PFK) has been purified 150-fold from potato tubers and the kinetic properties of the purified enzyme have been investigated both in the forward and the reverse direction. Saturation curves for fructose 6-phosphate and also for fructose 1,6-bisphosphate were sigmoidal whereas those for PPi and Pi were hyperbolic. In the presence of fructose 2,6-bisphosphate, the affinity for fructose 6-phosphate and for fructose 1,6-bisphosphate were greatly increased and the kinetics became Michaelian. The effect of fructose 2,6-bisphosphate was increased by the presence of fructose 6 phosphate and decreased by the presence of Pi. Consequently, the Ka for fructose 2,6-bisphosphate was as low as 5 nM for the forward reaction and reached 150 nM for the reverse reaction. On the basis of these properties, a procedure allowing one to measure fructose 2,6-bisphosphate in amounts lower than a picomole, is described. PMID- 6297886 TI - Messenger ribonucleoproteins of cells infected by simian virus 40 contain large T antigen. AB - In monkey or mouse cells undergoing lytic or transforming infection with simian virus 40, about 10% of large tumor antigen (T-antigen) molecules were consistently present in the cytoplasm and 90% in the nucleus. The bulk of cytoplasmic large T-antigen cosedimented in linear sucrose gradients with polyribosomes (150-500 S) and with messenger ribonucleoproteins sedimenting within 20-80 S. As determined by centrifugation in discontinuous sucrose gradients, its apparent density (1.2-1.3 g/ml) corresponded to that of ribonucleoproteins. In contrast, the bulk of nuclear T-antigen sedimented between 5-20 S and its apparent density (1.1 g/ml) corresponded to that of free protein. Nuclear T-antigen added before cell fractionation did not bind to cytoplasmic constituents. After dissociation of purified polyribosomes with puromycin/KCl or EDTA, cytoplasmic large T-antigen cosedimented with the released messenger ribonucleoproteins containing poly(A)-rich messenger RNA. Upon hydrolysis of the RNA with RNase A, large T-antigen exhibited the sedimentation properties and density of free protein. The results suggest that cytoplasmic large T-antigen is associated with messenger ribonucleoproteins. PMID- 6297887 TI - Regulation of the aminoacyl-tRNA synthetase complex of rat liver by phosphorylation/dephosphorylation in vitro and in vivo. PMID- 6297888 TI - The role of the proton-motive force and electron flow in light-driven solute transport in Rhodopseudomonas sphaeroides. AB - The initial rate of uptake of alanine was studied in whole cells of Rhodopseudomonas sphaeroides under conditions such that the proton-motive force generated by light-induced cyclic electron transfer, was composed of the membrane potential only. The proton-motive force was varied by varying the light intensity or by pretreatment of the cells with the uncoupler carbonylcyanide p trifluoromethoxyphenylhydrazone. At constant light intensity a threshold delta psi is required before alanine uptake occurs. Above this threshold value the rate of alanine uptake increases with the delta psi. The threshold value of the delta psi is higher the lower the light intensity. Under conditions of constant delta psi the rate of alanine uptake increases linearly with the light intensity. At low delta psi values a threshold light intensity is required which is higher the lower the delta psi. These results demonstrate that both a proton-motive force and electron transfer are required for alanine uptake. PMID- 6297889 TI - The complete amino-acid sequence of the low-spin class II cytochrome c-556 from Agrobacterium tumefaciens strain B2a. AB - The amino acid sequence of the soluble monohaem cytochrome c-556 from Agrobacterium tumefaciens, strain B2a, has been determined. The sequence was derived from peptides obtained by digestion of the apoprotein with trypsin and chymotrypsin, and by subdigestion of some of the peptides with Staphylococcus aureus protease and thermolysin. Sequencing of the various peptides was achieved by a combination of manual dansyl-Edman degradation and automatic liquid-phase sequence analysis. The main characteristic of this cytochrome is that the haem binding sequence Cys-Xaa-Yaa-Cys-His occurs in the C-terminal region of the polypeptide chain, the first cysteine being located 11 residues ahead of the C terminal lysine-122. As such, the protein belongs to cytochrome c sequence class II (sensu Ambler). The cytochrome c-556 is the first example known of a class II cytochrome of the low-spin type isolated from an obligate aerobic organism. PMID- 6297890 TI - Is cytochrome b really the antimycin-binding component of the cytochrome b--c1 complex of yeast mitochondria. AB - 1. The antimycin binding sites were found to be independent of cytochrome(s) b synthesis in pseudo-wild-type revertants of cytochrome b mutants. These revertants, whose primary mutation is located in the introns of the cob-box gene of mitochondrial DNA, have modified contents of cytochromes b-562 and b-565 and fully functional respiratory chain. 2. Missense mutations in three genetic loci allocated to the exons of the cytochrome b gene, abolish the strong affinity binding of antimycin. 3. It is proposed that the antimycin-binding component is not cytochrome b itself, but interacts with it in such a way that an alteration of the cytochrome b structure affects the antimycin-binding site. PMID- 6297891 TI - Kinetics of the reaction between human C1-esterase inhibitor and C1r or C1s. PMID- 6297892 TI - Isoenzymes of cAMP-dependent protein kinase in developing rat liver and in malignant hepatic tissues. AB - Total R proteins and total cAMP-dependent protein kinase activity during rat liver development reach highest values per unit DNA when the organ has attained full metabolic competence. The parallel changes indicate coordinate synthesis of R and C subunits during hepatic development. In contrast to total R2 . C2, protein kinase activation and endogenous cAMP levels were highest around birth. Immunotitration with anti-RI and anti-RII in the presence of protein-A-Sepharose of extracts obtained from various developmental stages and from hepatomas suggested a relation of both protein kinase I and II to the terminal differentiation of the organ rather than to cellular proliferation rates. The type-II enzyme appears to be subject to additional regulations connected with neonatal adaptation phenomena. A non-enzymic analysis of the protein kinase activation status is described. It is based on the determination of the ratio of amounts: R . cAMP/total R, which showed a linear correlation with the conventional protein kinase activity ratio (-cAMP/+cAMP). PMID- 6297893 TI - The role of pyrrolo-quinoline semiquinone forms in the mechanism of action of methanol dehydrogenase. PMID- 6297894 TI - Differential effect of tumor promoters on phorbol-ester-receptor binding and membrane fluorescence anisotropy in C3H 10T 1/2 cells. PMID- 6297895 TI - Interaction of a peripheral protein of the erythrocyte membrane, band 4.1, with phosphatidylserine-containing liposomes and erythrocyte inside-out vesicles. AB - Interactions of band 4.1 with mixed phospholipid membranes [phosphatidylserine (PtdSer), phosphatidylethanolamine, phosphatidylcholine, etc.] and erythrocyte inside-out vesicles were studied. Band 4.1 showed a higher affinity to PtdSer containing membranes. The amount of binding to PtdSer-containing liposomes was larger than that to PtdSer-lacking liposomes. The amount of binding to inside-out vesicles did not change significantly on a protease treatment of the vesicles. The amount of band 4.1 bound on inside-out vesicles decreased on PtdSer decarboxylase treatment of the vesicles. Ca2+ acted inhibitory to the binding of band 4.1. Band 4.1 together with PtdSer-containing vesicles but not with PtdSer lacking vesicles induced gelation of spectrin-actin copolymer solution. Ca2+ inhibited the gelation. Fluorescence energy transfer from PtdSer-containing vesicles to band 4.1 was larger than that from PtdSer-lacking vesicles. Band 4.1 caused a marked release of tempocholine from preloaded PtdSer-containing liposomes but not from PtdSer-lacking liposomes. The release was larger from liposomes containing more PtdSer. Ca2+ was inhibitory to the tempocholine release. We suggest from these results that band 4.1 provides another anchoring site for the cytoskeletal spectrin-actin network to PtdSer domains in the inner layer of erythrocyte membrane. This anchoring may be involved in functional regulation since the interaction causes the membrane permeability change that is dependent on Ca2+. PMID- 6297896 TI - Free N-acetylneuraminic acid in tissues in Salla disease and the enzymes involved in its metabolism. AB - Salla disease is a lysosomal storage disorder of unknown etiology, characterized biochemically by increased urinary excretion of N-acetylneuraminic acid. This compound has now been shown to occur in abnormally large amounts in liver and cultured skin fibroblasts from these patients. Quantification of N acetylneuraminic acid was performed using a new gas-chromatography/mass spectrometric single-ion method which is sensitive and specific. No abnormalities in the activity of several enzymes involved in sialic acid metabolism (N acetylneuraminate:pyruvate lyase, neuraminidase, CMP-N-acetylneuraminate N acylneuraminohydrolase and CTP:N-acyl-neuraminate cytidylyltransferase) were demonstrable. A possible explanation for the defect is a malfunctioning active transport of N-acetylneuraminic acid across the lysosomal membrane. PMID- 6297897 TI - Latent and active human polymorphonuclear leukocyte collagenases. Isolation, purification and characterisation. PMID- 6297898 TI - The collagenase inhibitor from human polymorphonuclear leukocytes. Isolation, purification and characterisation. PMID- 6297899 TI - Characterisation of beta 1-anticollagenase from human plasma and its reaction with polymorphonuclear leukocyte collagenase by disulfide/thiol interchange. AB - A beta 1-serum component, beta 1-anticollagenase, capable of inhibiting various mammalian tissue collagenases, was isolated from human plasma by gel filtration, affinity chromatography and ion-exchange chromatography. The inhibitor contains 1 2 free sulfhydryl groups, which are a prerequiste for inhibitory activity and for binding to the thiol-Sepharose affinity support. Alkylation of beta 1 anticollagenase by iodoacetamide blocks inhibitory activity. The inhibitor was purified to apparent homogeneity and exhibited a Mr = 30500 determined by sodium dodecyl sulfate/polyacrylamide gel electrophoresis. The amino acid and carbohydrate composition was determined. According to its composition and the isoelectric focussing beta 1-anticollagenase is an acidic protein with an isoelectric point of 5.6. Inhibition of human leukocyte collagenase proceeds in a strong 1 : 1 stoichiometric reaction. The mechanism of this association takes place by a disulfide/thiol interchange reaction as has been previously indicated for human leukocyte collagenases in forming the latent enzyme [Macartney, H. W. and Tschesche, H. (1980) FEBS Lett. 119, 327-332]. The beta 1-anticollagenase- leukocyte-collagenase complex (latent enzyme) is activatable by disulfide containing compounds such as cystine, oxidised glutathione, insulin, relaxin, trypsinogen and others, but not by 179,203-di(S-carboxymethyl)trypsinogen, or its trypsin derivative. Compounds containing inaccessible disulfide bonds, e.g. chymotrypsin, or sulfhydryl groups, e.g. D-penicillamine, do not activate the complex. Activation is, however, easily obtained with the oxidised-glutathione generating system myeloperoxidase/H2O2/glutathione as was previously demonstrated for the human leukocyte latent collagenase activatable in a phagocytosis simulated respiratory burst [Tschesche, H. and Macartney, H. W. (1981) Eur. J. Biochem. 120, 183-190]. PMID- 6297900 TI - Kinetics of reduction of putidamonooxin by NADH-putidamonooxin oxidoreductase, sodium dithionite and superoxide radicals. PMID- 6297901 TI - Demonstration and characterisation of a testicular receptor for 1,25 dihydroxycholecalciferol in the rat. PMID- 6297903 TI - Rotational motion of actin monomer at low and high salt concentration. PMID- 6297902 TI - Two proteolytic degradation products of calf-thymus poly(ADP-ribose) polymerase are efficient ADP-ribose acceptors. Implications for polymerase architecture and the automodification of the polymerase. AB - Two polypeptides with molecular masses of 76 and 59 kDa were found to copurify with poly(ADP-ribose) polymerase from calf thymus, and to be as efficient acceptors of ADP-ribose as the polymerase itself. Analysis of their CNBr fragments by sodium dodecylsulfate/polyacrylamide gel electrophoresis revealed that the polypeptides were derived from the 112-kDa polymerase. Isolation of poly(ADP-ribose) polymerase in the absence of protease inhibitors resulted in a loss of more than 90% of the polymerase activity and an increased proportion of the 76-kDa and 59-kDa polypeptides in the final polymerase preparation. When the polymerase and the two polypeptides were separated by gel filtration or polyacrylamide gel electrophoresis in 5% acetic acid, no polymerase activity was found associated with the two fragments. Analysis of the CNBr fragments of the three polypeptides after incubation of the enzyme preparation with [32P]NAD showed that most of the fragments were radioactive, indicating multiple ADP ribosylation sites. Several ADP-ribosylated fragments were found to be common to all three polypeptides, or to two of them. PMID- 6297904 TI - The distribution of redox equivalents in the anaerobic respiratory chain of Paracoccus denitrificans. PMID- 6297905 TI - Determination of protonation sites in thermospermine and in some other polyamines by 15N and 13C nuclear magnetic resonance spectroscopy. AB - The pK values of thermospermine, a novel asymmetric tetraamine (4,8-diazadodecane 1,12-diamine), have been determined for the first time by the conventional titration method with subsequent data analyses. It was found to be the most basic polyamine among naturally occurring tetraamines. The 15N NMR titration method was applied to determine its protonation site. Natural-abundance 15N magnetic resonance spectra of the polyamine have been recorded as a function of pH from its free to the protonated form. Its apparent pK values, as well as the intrinsic chemical shifts for each stage of protonation, have been calculated from the NMR titration curve by using a nonlinear least-squares method. The four protonation sites of thermospermine have been successfully determined to be N12, N1, N-8, and N-4 in that order from the free to the completely protonated form. In a similar manner the protonation sites of spermidine and two symmetric tetraamines, thermine and spermine, have been established. 13C NMR experiments for the four polyamines were almost consistent with those of 15N NMR. Owing to the much higher concentration of the polyamines furnished for NMR titration experiments, the pK values obtained by NMR were larger than those derived by the conventional titration methods. PMID- 6297906 TI - Effects of experimental insulin-dependent diabetes on the beta-adrenergic receptor-coupled adenylate-cyclase system and lipolysis in fat cells of the rat. PMID- 6297908 TI - The influence of the ionic conductance on the relation between electron transport and proton-motive force in intact cells of Rhodopseudomonas capsulata. AB - 1. The dependence of membrane potential (delta psi) on the rate of respiration in darkened intact cell suspensions of Rhodopseudomonas capsulata was distinctly non linear: severe inhibition of respiration with either rotenone or KCN led to only a small drop in delta psi. 2. In the presence of 0.3 microMs carbonylcyanide p trifluoromethoxyphenylhydrazone [CF3OPhzC(CN)2] the dependence of delta psi on respiratory rate became linear. Consequently, and particularly at lower concentrations of CF3OPhzC(CN)2, there was a pronounced, synergistic depression of the respiratory delta psi with CF3OPhzC(CN)2 and either rotenone or KCN. 3. Antimycin A, at a concentration which strongly inhibited the photosynthetic electron transport chain, only slightly lowered the light-induced delta psi in anaerobic cell suspensions. Antimycin and CF3OPhzC(CN)2 synergistically lowered delta psi generated by illumination. 4. The light-induced delta psi in anaerobic cells was only about 1.5-times larger than the respiratory-induced delta psi in darkened cells. Nevertheless it required approximately 16-times more CF3OPhzC(CN)2 to collapse the photosynthetic delta psi than the respiratory delta psi. 5. These results are discussed with reference to the ionic current/delta psi relation described in [J.B. Jackson (1982) FEBS Lett. 139, 139-143]. The unifying feature is that the intrinsic conductance of the cell membrane is strongly dependent on delta psi but the conductance due to CF3OPhzC(CN)2 is independent of delta psi. PMID- 6297907 TI - Desulfovibrio Gigas hydrogenase: redox properties of the nickel and iron-sulfur centers. AB - Below 30 K, oxidized Desulfovibrio gigas hydrogenase presents an intense electron paramagnetic resonance (EPR) signal centered at g = 2.02, typical of an iron sulfur center. In addition a rhombic EPR signal, attributed to Ni(III) species, is also observed [LeGall, J., Ljungdahl, P., Moura, I., Peck, H.D., Jr, Xavier, A.V., Moura, J.J.G., Teixeira, M., Huynh, B.H., and DerVartanian, D.V. (1982) Biochem. Biophys. Res. Commun. 106, 610-616; and Cammack, R., Patil, D., Aguirre, R., and Hatchikian, E.C., (1982) FEBS Lett. 142, 289-292]. At higher temperatures (77 K) the iron-sulfur EPR signal is broader and all the EPR features of the rhombic nickel signal can easily be observed. We have now obtained additional information concerning the redox properties of these EPR active centers, using an EPR redox titration method in the presence of dye mediators at pH = 8.5. The mid point potential was determined to be -70 mV for the Fe,S cluster and -220 mV for the Ni center. Intermediate oxidation states were obtained upon partial reduction with either dithionite or hydrogen. Although upon dithionite reduction the centers are reduced in the order of decreasing mid-point reduction potentials, under a hydrogen atmosphere the nickel center reduces preferentially. This suggests a catalytic involvement of the nickel redox center in the binding of hydrogen. Preliminary Mossbauer studies on Desulfovibrio gigas hydrogenase reveal the presence of a paramagnetic 3 Fe center and two 4 Fe centers. The 3 Fe center is responsible for the g = 2.02 EPR signal but the two 4 Fe centers have been so far undetectable by EPR. PMID- 6297909 TI - Characterization of phosphorylated and native cGMP-dependent protein kinase. PMID- 6297910 TI - Occurrence of an inhibitory guanine nucleotide-binding regulatory component of the adenylate cyclase system in cyc- variants of S49 lymphoma cells. PMID- 6297911 TI - The biphasic action of cardiac glycosides on the Na+, K+-pump and its relevance in the treatment of heart failure. PMID- 6297912 TI - Transferrin receptors and gallium-67 uptake in vitro. AB - The relationship was studied between the number of transferrin-receptor positive cells and in vitro uptake of 67Ga and 125I-labeled transferrin in human cell lines, including two normal cell lines (WI-38 and foreskin fibroblasts), two transformed cell lines (AV-3 amnionic cells and Chang liver cells), and two neoplastic cell lines (HEp-2, larynx cancer and HeLa, cervical cancer). Transferrin receptors were determined by an indirect immunofluorescence technique based on their ability to bind purified human transferrin. Gallium-67 uptake was determined after a 24-h incubation of cells with Ga-67 in the presence and absence of transferrin (0-2.5 mg/ml). 125I-labeled transferrin uptake was also obtained after a 24-h incubation. The fraction of cells with transferrin receptors was low in normal cell lines (3% and 9%), intermediate with transformed cell lines (33% and 61%). Transferrin stimulated 67Ga uptake by all six cell lines. However, there was poor correlation between the number of transferrin receptor positive cells and 67Ga uptake either in the presence (r = 0.21) or absence (r = 0.46) of human transferrin. Likewise, there was poor correlation between the number of transferrin-receptor positive cells and 125I-labeled transferrin uptake (r = 0.35). In contrast, the correlation between 67Ga uptake (in the presence of transferrin) and 125I-labeled transferrin uptake was highly significant (r = 0.96). These results suggest that human cell lines in culture are capable of both transferrin-dependent and transferrin-independent uptake of 67Ga. PMID- 6297914 TI - Properties of a Herpes virus-transformed hamster cell line--I. Growth and culture characteristics of sublines of high and low metastatic potential. AB - A Herpes virus hominis type 2-induced tumour line of the Syrian hamster, designated HSV-2-333-2-26, exhibited a low level of spontaneous metastasis from the primary subcutaneous tumour mass. Two lung foci, met A and met B, were resected and transplanted s.c.; both lines showed an elevated metastatic potential compared with the parent cell line such that all animals inoculated with these cells developed metastatic disease within 40 days after resection of the primary tumour. Neither the rapidity of cell growth in vivo or in vitro, nor anchorage-independent growth or degree of attachment to a solid substrate in vitro are implicated in the altered behaviour of the metastatic cell lines. PMID- 6297913 TI - Intermittent ketoconazole therapy of chronic mucocutaneous candidiasis in childhood. AB - We report the clinical and laboratory findings in two children with chronic mucocutaneous candidiasis (CMC) treated successfully with intermittent long-term ketoconazole therapy. Both had chronic infection of the nails, skin and mucous membranes with positive cultures for candida. Both were resistant to multiple local and systemic antifungal agents. After institution of ketoconazole therapy there was a dramatic improvement with clearing of the oral (one week), skin (two months) and nail lesions (5 months). After 8 months the drug was stopped and clinical remission persisted for 10 and 7 months respectively. Relapse of oral candidiasis was treated with a short course of ketoconazole (4-16 weeks) leading to complete healing of the lesions. Clinical improvement was not related to an amelioration in lymphocyte transformation. There was no change in the progressive deterioration of the lymphocyte responses to candida antigen which was probably due to persisting candida cell wall components (e.g. mannan). PMID- 6297915 TI - Oral high-dose medroxyprogesterone acetate causes adrenal suppression in patients with breast cancer. PMID- 6297916 TI - Bone marrow involvement in adult soft tissue sarcomas. AB - Bone marrow biopsy was performed as part of the initial assessment in 74 patients with soft tissue sarcoma. Infiltration of the marrow by tumour was present in four cases, all from the group of 56 patients who had other evidence of metastatic disease, giving an overall incidence of 7%. The histological subtypes were pleomorphic rhabdomyosarcoma, angiosarcoma, synovial sarcoma and myxoid liposarcoma, of which the first three were high-grade tumours. Although it was not possible to determine whether response to chemotherapy was influenced by marrow involvement, haematological toxicity seemed excessive. PMID- 6297917 TI - Prolactin receptors in human breast cancer. AB - Prolactin receptors have been measured in 92 human breast carcinomas. Both free and total receptors (after desaturation by MgCl2) have been looked for. Free receptors have been found in 46% of the cases, total receptors in 72%. Specific binding ranges from 0.8 to 8.0%. No correlation could be found between prolactin receptors and age, weight, menopausal status and pathological features (differentiation, histoprognostic grading, cellular density). A highly significant correlation has been found between prolactin receptors on the one hand and estradiol and progesterone receptors on the other. PMID- 6297918 TI - Host endocrine status mediates oncogenesis: leukemia virus-induced carcinomas and reticulum cell sarcomas in acyclic or normal mice. AB - A highly selective leukemogenic virus obtained from thymic lymphosarcomas (LS) of C57BL/6 mice was inoculated intraperitoneally into normal or gonadectomized C57BL/6 male mice and into normal or masculinized, acyclic, Charles River (CR) outbred female mice. This virus induced early onset of LS in both normal and gonadectomized C57BL/6 mice. The same virus induced a relatively late onset of many mammary adenocarcinomas in the normal CR mice and ovary adenocarcinomas and systemic neoplasms (LS and reticulum cell neoplasms) in the acyclic females. Topography, histopathology and the average latent period of the tumors were completely different between the normal and the acyclic females. This is consistent with the idea that a leukemia virus can induce tumors other than leukemia and that permanent alterations of the physiological hormonal cyclicity plays a major role in determining whether or on which target cells the ubiquitous virus will exert its carcinogenic action. PMID- 6297919 TI - Cytotoxic T cell recognition of Epstein-Barr virus-infected B cells. III. Establishment of HLA-restricted cytotoxic T cell lines using interleukin 2. AB - Epstein-Barr virus (EBV)-specific cytotoxic T cell precursors, present in the circulation of previously infected (seropositive) individuals, have been reactivated in vitro by challenging with autologous EBV-transformed cells, and the reactivated populations subsequently expanded as interleukin 2 (IL2) dependent cell lines. These lines were dominated by T cells possessing the cytotoxic/suppressor cell surface phenotype and, when tested for effector function in chromium-release assays, demonstrated potent EBV-specific, HLA-A and B antigen-restricted cytotoxicity even when derived from seropositive donors whose initial cytotoxic response to in vitro reactivation was relatively weak. With all the lines tested from 10 seropositive donors, strong killing of autologous EBV-transformed cells was observed in the absence of any significant lysis of autologous mitogen-stimulated lymphoblasts or of a panel of EBV genome negative cell lines sensitive to natural killing. Furthermore, the availability of IL2-expanded effectors cell populations allowed their being tested upon a wide panel of allogeneic EBV-transformed targets such that the dominant HLA-restricted reactivities within these populations could be identified. Monoclonal antibody blocking experiments confirmed that lysis of the autologous EBV-transformed cell line by IL2-expanded effectors could be specifically inhibited (a) by pretreatment of the target cells with antibodies binding to the HLA/beta 2 microglobulin complex, and (b) by pretreatment of the effector cells with the cytotoxic/suppressor T cell-specific antibody Leu 2a. PMID- 6297920 TI - Sendai virus-specific T cell clones II. Induction of interferon production by Sendai virus-specific T helper cell clones. AB - Interferon (IFN) was detected upon co-culture of cloned Sendai virus (SV) specific T lymphocyte with SV-presenting syngeneic stimulator cells. The antiviral activity was defined as IFN-alpha, beta. The T cell clones, upon contact with antigen-presenting stimulator cells, stimulated adherent cells present in the stimulator cell population to secrete IFN. Induction of IFN production was independent from interleukin 2 production by the T cell clones. PMID- 6297921 TI - Superoxide anion production from mouse peritoneal macrophages stimulated with surface-bound IgG and immune complexes: adsorption characteristics of IgG in relationship to biological activity. AB - The bridging of IgG antibody molecules by either multivalent antigen or protein A adsorbed to a polymer surface triggers superoxide anion (O-2) generation from mouse peritoneal macrophages. Macrophages, upon stimulation with IgG adsorbed to a hydrophobic polymer, polystyrene, also significantly generated O-2. In contrast, soluble IgG did not stimulate macrophages. From these results we speculated that IgG adsorbed to polystyrene produces the same kind of a conformational change in its Fc region as seen in immune complexes, and leads to enhancement of the ability of the Fc region to bind macrophage Fc receptors, acting as a multivalent cross-linking agent to aggregate the Fc receptors. The surface charge density of the immunosorbent did not affect the antigen recognition function of antibody, but markedly affected its effector function. These findings suggest that activation of the Fc receptor-mediated function of macrophages requires, on the one hand, optimal conformational change and orientation of IgG and immune complexes adsorbed to the polymer surface and, on the other hand, the bridging of IgG molecules by multivalent agents. PMID- 6297922 TI - Coronary vasoconstriction induced by leukotrienes in the anaesthetized dog. AB - In the anaesthetized dog LTC4, LTD4 (0.3-10 micrograms, injected into the coronary artery) and the thromboxane-mimetic U46619 (5-10 micrograms) decreased coronary blood flow. LTE4 (1-10 micrograms), however, did not affect coronary blood flow. The vasoconstrictor responses to LTC4, LTD4 or U46619 were not altered by the cyclo-oxygenase inhibitor indomethacin (5 mg/kg i.v.). LTC4 and LTD4 did not stimulate the release of any prostaglandin-like substance into the pericardial fluid. It is concluded that LTC4 and LTD4 are able to produce coronary vasoconstriction in vivo independent of the production of any cyclo oxygenase metabolites of arachidonic acid. PMID- 6297923 TI - Serotonergic denervation changes binding characteristics of beta-adrenoceptors in rat cortex. PMID- 6297924 TI - [3H]nitrendipine labelling of the Ca2+ channel in skeletal muscle. PMID- 6297925 TI - Ontogeny of alpha 1-adrenergic receptors in the rat myocardium: effects of hypothyroidism. AB - The effects of propylthiouracil (PTU) treatment on alpha 1- and beta-adrenergic receptors in neonatal and adult rat myocardium were examined with the alpha 1 selective antagonist [3H]prazosin and beta-antagonist (-)-[3H]dihydroalprenolol ((-)-[3H]DHA). Pre and postnatal PTU treatment (congenital hypothyroid state) decreased ventricular size and protein content at 15 and at 28 days (P less than 0.01). Similarly PTU treatment after maturation (acquired hypothyroid state) is associated with decreased ventricular size and protein content (P less than 0.01). PTU treatment did not alter ventricular DNA content at any age examined as compared to euthyroid controls. Specific [3H]prazosin binding to ventricular membrane increased with advancing postnatal age in both congenital hypothyroid pups and controls, reaching peak receptor density at 15 days of postnatal age (106 +/- 8 vs. 151 +/- 7 fmol X mg-1 protein). The density at 15 and 28 days was however significantly lower in hypothyroid pups (P less than 0.01, P less than 0.02). The effect of congenital hypothyroidism on (-)-[3H]DHA binding was more marked which significantly decreased from the first postnatal day. Acquired hypothyroidism is also associated with a decreased density of alpha 1- and beta adrenergic receptors in rat myocardium. We conclude that alpha 1- and beta adrenergic receptors are modulated by thyroid hormone in developmental as well as mature stages of the rat. PMID- 6297926 TI - Role of 5-HT and NA in spinal dopaminergic analgesia. AB - Spinal rats and rats with an intact neuraxis given the dopamine (DA) agonist R apomorphine (0.31-1.75 mumol/kg) in the lumbar subarachnoid space by intrathecal injection were tested 5 and 10 min later for spinal analgesia (increased tail flick response latency). Apomorphine produced analgesia in spinal rats but not in rats with an intact neuraxis. However, pretreatment of intact rats with the serotonergic (5-HT) receptor antagonist methysergide, the noradrenergic (NA) receptor antagonist phentolamine or the two antagonists together led to a dose dependent analgesia following apomorphine. Intact rats pretreated with the monoamine depleting drug reserpine, the 5-HT synthesis inhibitor p chlorophenylalanine (PCPA) or the NA synthesis inhibitor FLA 63 also showed analgesia to apomorphine. On the other hand, pretreatment with the catecholamine depleting agent alpha-methyl-p-tyrosine (AMPT), the beta-adrenergic receptor blocker propranolol or the opioid receptor antagonist naloxone failed to produce DA analgesia. The present findings suggest that both 5-HT and NA descending fiber systems exert tonic inhibitory effects on spinal DA nociceptive processes. PMID- 6297927 TI - Effects of morphine on net water absorption, mucosal adenylate cyclase activity and PGE2 metabolism in rat intestine. AB - Morphine (10 mg/kg s.c.) was shown to increase net basal water and electrolyte absorption in rat ileum and colon perfused in vivo. These effects were significantly greater in the ileum. Under the same conditions of dosage and time course of administration the activities of relevant mucosal enzymes were studied. Ileal mucosal adenylate cyclase stimulated by prostaglandin E2 (PGE2) in vitro was inhibited by morphine pretreatment. Colonic stimulated cyclase activity was unaffected as were basal activities in both regions. Morphine did not alter the activities of enzymes involved in PGE2 metabolism and is unlikely to act by modulating endogenous PG levels. Transport ATPase activities, were also unchanged in morphine treated animals. These results are discussed in relation to the mechanism of action of morphine on intestinal mucosa. PMID- 6297928 TI - Action of stereoisomers of (imidazolinyl-2)-2-benzodioxane-1-4 or 2-(2-(1,4 benzodioxanyl))-2-imidazoline (170 150; RX 781094) on peripheral presynaptic and central alpha 2-adrenoceptors. AB - In the vas deferens of the rat, (-)-170 150 (imidazolinyl-2)-benzodioxane-1-4 or 2-(2-(1,4-benzodioxanyl))-2-imidazoline appeared to be more selective than (+) 170 150 for antagonizing the inhibitory effects of clonidine on the twitch response. In contrast, (+)-170 150 was the most potent of the two isomers for antagonizing the centrally mediated clonidine- or azepexole-induced sleep in chickens. These unexpected results seem to indicate that there are several subpopulations of alpha 2-adrenoceptors. PMID- 6297929 TI - GABAA- and GABAB-receptor-mediated modification of intestinal motility. AB - The actions of gamma-aminobutyric acid (GABA) and its analogues, 3-amino-1 propanesulphonic acid (3APS) and baclofen, have been investigated using isolated segments of the guinea-pig ileum and distal colon. GABA and 3APS, but not baclofen, induced GABAA-receptor mediated effects; prompt, dose-dependent contractions of the ileum which were antagonised by bicuculline, picrotoxinin, piretanide, tetramethylenedisulphotetramine, atropine and tetrodotoxin. Baclofen and GABA, but not 3APS, induced a dose-dependent GABAB-receptor mediated depression of electrically elicited twitch contractions of the ileum, unaffected by the GABAA-receptor antagonists or by antagonism of adenosine, adrenergic, opiate or nicotinic receptors. In the distal colon, baclofen and GABA caused a bicuculline- and picrotoxinin-insensitive depression of spontaneous cholinergic contractions. Desensitization to GABA and baclofen, and cross-desensitization to both agonists was observed. Combined antagonism of GABAA-receptors and desensitization to baclofen slowed pellet expulsion to the same extent as GABA desensitization alone, indicating that both GABAA- and GABAB-receptor sites are involved in this modification of peristalsis by GABA. PMID- 6297930 TI - Dopamine receptor profile of co-dergocrine (Hydergine) and its components. AB - Co-dergocrine (Hydergine), an ergot preparation composed of four dihydrogenated peptide ergot alkaloids (dihydroergocornine, dihydroergocristine, dihydro-alpha ergokryptine, dihydro-beta-ergokryptine, 3:3:2:1), has been reported to exert in vivo effects suggesting an interaction with dopaminergic systems. The present investigation provides evidence that, in the striatum of the rat, co-dergocrine and its components interact directly with D1- and D2-subtypes of dopamine receptors. In homogenates of rat striatum, co-dergocrine and three of its components (dihydroergocornine, dihydro-alpha-ergokryptine, dihydro-beta ergokryptine) stimulate cyclic AMP formation (D1-receptor response) having similar EC50 values but different efficacies. The same compounds inhibit electrically evoked tritium overflow from rat striatal slices preincubated with [3H]choline (D2-receptor response) at about 50 times lower concentrations. Here again the compounds exhibit differential maximal effects. One component, dihydroergocristine, antagonises both receptor types. The effect of co-dergocrine in functional responses mediated by both D1- and D2-receptors seems to reflect the summation of the contribution of its components. PMID- 6297931 TI - Brain catecholamine metabolites and behavior in morphine withdrawal. AB - Morphine withdrawal behavior, brain and plasma catecholamine metabolites, and brain beta-noradrenergic receptor binding were examined after acute treatment with naloxone in rats treated with morphine pellets or a sham pelleting procedure. Increases in brain 3-methoxy-4-hydroxyphenethyleneglycol (MHPG), a norepinephrine metabolite, occurred in parallel with rated withdrawal behavior. Withdrawal behavior correlated significantly with brain, and, more modestly, with plasma levels of MHPG but did not correlate with beta-receptor binding or HVA. The effectiveness of debrisoquin sulfate was variable, but the reductions in withdrawal signs and cerebral cortex MHPG were strongly correlated. These data support a direct relationship between presynaptic noradrenergic hyperactivity and opiate withdrawal behavior. PMID- 6297932 TI - Contractile activities of several cysteine-containing leukotrienes in the guinea pig lung strip. AB - Eight biosynthetically formed cysteine-containing leukotrienes dose dependently (0.01-100 nM) contracted parenchymal strips of guinea-pig lung. The response to the leukotrienes was slow in onset, remarkably long-lasting and often tachyphylactic upon repeated administration. All the leukotrienes (LTC3, 8,9 LTC3, LTC4, 11-trans-LTC4, LTC5, LTD4, LTE4 and 11-trans-LTE4) were equally active full agonists for contraction, but approximately 5000 times as potent as histamine on a molar basis. Radiolabelled leukotriene C was insignificantly metabolized during the assay of contractile activity, indicating that the leukotrienes were active per se. The equipotency of LTC4, LTD4 and LTE4 also indicates that each of these three major constituents of slow reacting substance of anaphylaxis (SRS-A), may be considered a significant mediator of allergen induced bronchoconstriction. In addition, the closely similar contractile activity of glutathionyl-substituted leukotrienes derived from polyunsaturated fatty acids other than arachidonic acid, suggests that they may contribute to bronchospastic disease under pathological and nutritional conditions promoting their formation. PMID- 6297933 TI - Inactivation of GABA receptors by phenoxybenzamine: effects on GABA-stimulated benzodiazepine binding in the central nervous system. AB - Treatment of particulate fractions of rat forebrain with phenoxybenzamine produced dose related inhibition of specific muscimol binding. Scatchard analysis revealed a significant reduction of binding sites but no change in affinity constant. The inhibition was not reversed by washing or incubation at 37 degrees C suggesting that phenoxybenzamine alkylates GABA receptors, i.e. acts as a non competitive, non-equilibrium inhibitor. Phenoxybenzamine inhibited the GABA induced increase in specific diazepam binding but had little direct effect on diazepam binding. Preincubation of synaptic membranes with GABA or muscimol, but not diazepam, protected the muscimol binding site from inactivation by phenoxybenzamine. PMID- 6297934 TI - Increase of [3H]clonidine binding sites induced by adenosine receptor agonists in rat vas deferens in vitro. AB - In vitro binding experiments were done to study the effects of adenosine receptor agonists on induction of [3H]clonidine binding sites in rat vasa deferentia by reserpine. In the presence of 1 microM adenosine and 2-chloroadenosine, respectively, the specific binding of [3H]clonidine to vas deferens homogenates of reserpine-treated rats increased 38 and 42% over the control value. The increase in binding sites induced by adenosine was inhibited by the adenosine receptor antagonist theophylline. No effect of 1 microM 2-chloroadenosine on the beta-adrenoceptor or alpha 1-adrenoceptor was detected. The results imply a direct interaction between alpha 2-adrenoceptor and adenosine receptor and suggest that stimulated adenosine receptors unmasked [3H]clonidine binding sites in membranes of rat vas deferens. PMID- 6297935 TI - Benzodiazepine receptor sites in bovine pineal. AB - [3H]Flunitrazepam (FNZP) binds to a single class of high affinity binding sites in bovine pineal membranes (KD 2.8 nM). FNZP binding increased by 25% after exposure to gamma-aminobutyric acid and exhibits a high degree of specificity for drugs acting on the central type of benzodiazepine receptor. High affinity binding of [3H]ethyl-3-carboline ester to pineal membranes has a KD 0.8 nM and about 60% of the maximal number of FNZP binding sites. PMID- 6297936 TI - The use of the slowly degradable analog, alpha, beta-methylene ATP, to produce desensitisation of the P2-purinoceptor: effect on non-adrenergic, non-cholinergic responses of the guinea-pig urinary bladder. AB - alpha, beta-Methylene ATP has advantages over ATP in producing desensitisation of the P2-purinoceptor since it is degraded more slowly than ATP and does not initiate synthesis of prostaglandins. Following desensitisation of the excitatory P2-purinoceptors in the guinea-pig urinary bladder, the excitatory responses to non-adrenergic, non-cholinergic nerve stimulation were abolished, while those to acetylcholine and histamine were little affected. This result is consistent with the purinergic nerve hypothesis. PMID- 6297937 TI - Tritium-film autoradiography of sodium-independent glutamate binding sites in rat brain. PMID- 6297938 TI - Embryonal carcinoma cell growth and differentiation. Production of and response to molecules with transforming growth factor activity. AB - Transforming growth factors are known to induce anchorage-independent growth of non-transformed cells, and are released by a variety of cells, including MSV transformed cells. This study demonstrates that the differentiated cells derived from F9 and PC-13 embryonal carcinoma cells, but not the parental cells themselves, respond by increased growth to several factors released by MSV transformed cells, including partially purified sarcoma growth factor. The chemical properties of the growth-promoting activity are shown to match the chemical properties of the transforming growth factors released by MSV transformed cells. Furthermore, F9 and PC-13 embryonal carcinoma cells, which do not respond to factors released by MSV-transformed cells, are shown to release factors with transforming growth factor activity. Based on the close relationship between mouse embryonal carcinoma cells and cells of early mouse embryos, it is suggested that molecules with transforming growth factor activity may play a role during the early stages of mammalian development. PMID- 6297939 TI - Architecture and dynamics of microvillar photoreceptor membranes of a cephalopod. AB - Microvillar membranes of cephalopod photoreceptors, Eledone aldrovandii were analysed with respect to their protein and lipid composition. Molecular dynamics of this membrane type were investigated by ESR measurements using frog rod outer segment membranes as a reference system. The photoreceptor membrane is composed of about 56 wt% protein and 44 wt% lipid. Rhodopsin (mol wt 51 000) represents at least 70% of the membrane protein. The molar ratio of phospholipid to cholesterol to rhodopsin is about 55:24:1. Phosphatidylcholine (28.9 mol%) and phosphatidylethanolamine (27.8 mol%) are the major phospholipids. The ESR measurements suggest that the cephalopod photoreceptor membrane is less fluid than from rod outer segment membranes although the major phospholipids show remarkable high levels of polyunsaturated fatty acids (e.g. 88 mol% in phosphatidylethanolamine). It is concluded that the lower fluidity of microvillar membranes results in part from high cholesterol content and that a restricted mobility of rhodopsin in this membrane does not result only from the fact that the membrane is rolled into a microvillar structure. PMID- 6297940 TI - Selective solubilization of two populations of polypeptides from bovine retinal basement membranes. AB - Basement membranes isolated from the entire neural retinae of normal cows consist of a heterogeneous mixture of polypeptides resolvable as two major populations, one collagenous and the other non-collagenous. The non-collagenous population consists of disulfide-bonded aggregates of which the major fraction is soluble in lithium dodecyl sulfate (LDS) at 4 degrees C without reducing agent, with the remainder requiring reduction for solubilization at 4 degrees C. Only the second 4 degrees C extract is contaminated with small amounts of hydroxylated amino acids. The non-collagenous population consists of up to 25 polypeptides detectable in sodium dodecyl sulfate (SDS) gels, all of which stain blue with Coomassie Blue R-250, are insensitive to bacterial collagenase and migrate on or slightly below the central diagonal in urea-SDS/SDS electrophoresis. The collagenous population is soluble in LDS at 100 degrees C under reducing conditions and has a collagenous amino acid composition, although consisting of only 272 glycine residues/1000. This fraction consists of four major and four minor polypeptides detectable in SDS gels, all of which stain red (metachromatically) with Coomassie Blue R-250, are degraded by bacterial collagenase and migrate below the central diagonal in urea-SDS/SDS electrophoresis. The four major collagenous polypeptides are larger than the alpha-chains of type I collagen. In total, the solubilized proteins account for 60% of the general protein, but only 30% of the sum of hydroxyproline and hydroxylysine. The insoluble residue has a collagenous amino acid composition similar to that of the 100 degrees C extract, but with lower 3-hydroxyproline and hydroxylysine contents. PMID- 6297941 TI - Biochemical events associated with lymphocyte activation in aging. K+ transport and sensitivity of the Na+-K+ pump to digoxine. AB - The present investigation was directed toward answering the question of whether some age-related changes of membrane dependent triggering mechanisms during lymphocyte activation could account for the depressed T cell response to mitogens in aging. For this purpose, the K+ movements were analyzed in PHA-stimulated peripheral blood lymphocytes (PHA-PBL) from old humans (O) compared to adult (A). Indeed, plasma membrane Na+, K+, ATPase activation plays an essential role in cell proliferation and results from direct interaction between the loaded mitogen receptor and the enzyme. No difference could be found in the magnitude and the timing of the PHA-induced increase of K+ fluxes between PHA-PBL from O and A despite a higher K+ inflow in unstimulated but 20-hour preincubated PBL from O. Further experiments showed that the lectin-induced triggering mechanism of cation transport resulted from digoxine (DGX: a glycosid cardiotonic) sensitive ATPase. Moreover, whereas PBL from O exhibited a decreased PHA-induced DNA synthesis, DGX depressed the thymidine incorporation by 72-hour cultured PHA-PBL within the same inhibitory dose-related pattern in both O and A. We conclude that the triggering mechanism of Na+, K+-ATPase induced by PHA occurs adequately in early stimulated PBL from old subjects. In addition, digoxine sensitive structures work freely during PHA-induced lymphocyte proliferation in aging, thereby supporting further arguments for adequate Na+, K+-ATPase activity. PMID- 6297942 TI - Role of erythropoietin and cyclic nucleotides in erythroid cell proliferation in fetal liver. AB - Cyclic AMP and cyclic GMP have been implicated as regulators of proliferation and differentiation in a variety of tissues including erythroid cells. Present studies indicate that erythropoietin is capable of stimulating an accumulation of cyclic GMP in fetal liver cultures. Erythropoietin (Ep) decreases the duration of the G1 phase of the cell cycle, diminishing the total cell cycle time by approximately 40%. In proliferating cells to which no Ep was added, concentrations of cyclic AMP and cyclic GMP changed as cells progressed through the cycle. Increases in cyclic GMP levels are associated with DNA synthesis and mitosis. On the other hand, cyclic AMP levels are associated with the G1 and G2 phases of the cycle. When Ep was included in cultures, the same cyclic nucleotide pattern relative to phases of the cell cycle was maintained. Thus, it is likely that cyclic GMP and cyclic AMP exert a regulatory influence on proliferation and differentiation of developing erythroid cells. PMID- 6297943 TI - 67Ga scintigraphy, serum lysozyme and angiotensin-converting enzyme in pulmonary sarcoidosis. AB - 67Ga scintigraphy, using a 67Ga accumulation score, was compared with serum lysozyme (LZM) and angiotensin-converting enzyme (ACE) levels in 34 patients with biopsy-proven, pulmonary sarcoidosis. Serum LZM and ACE values varied between 1.0 to 21.3 mg/l (mean 4.3 +/- 3.4 mg/l) and 33 to 146 U/l (mean 69 +/- 26 U/l), respectively. Normal values were found in 26 and 35% of the patients. All patients, however, had abnormal 67Ga uptake in the pulmonary hilar lymph nodes and/or parenchyma. The follow-up of untreated and treated patients supports the suggestion that 67Ga scintigraphy is more useful for assessing the extent and activity of the intrathoracic sarcoid lesions. Serum LZM and ACE measurements are helpful, but normal LZM and ACE values do not exclude activity and progression of disease in pulmonary sarcoidosis. PMID- 6297944 TI - Granular cell myoblastoma of the bronchus. AB - A case is described in which a right-sided bronchus tumour was found incidentally at endoscopic examination for left-sided pneumonia. No clinical or radiographic signs of this bronchial mass were present. Endoscopic biopsy proved the tumour to be a granular cell myoblastoma. PMID- 6297945 TI - Possible mechanisms in asthma. PMID- 6297946 TI - Bronchial reactivity in relation to occupational bronchitis and asthma. PMID- 6297947 TI - Isocyanate induced asthma. PMID- 6297948 TI - Mechanisms of isocyanate induced asthma. PMID- 6297949 TI - Long-term respiratory effects of isocyanate exposure. PMID- 6297950 TI - Non-specific bronchial hyper-reactivity in workers exposed to toluene di isocyanate, diphenyl methane di-isocyanate and colophony. AB - Non-specific bronchial reactivity to histamine has been measured before specific occupational bronchial tests in the following groups: 51 workers exposed to toluene di-isocyanate (TDI); 40 workers exposed to diphenylmethane diisocyanate (MDI); 45 electronics workers exposed to colophony fumes and 13 unexposed controls. Finally 38 electronics workers had repeated measurements after moving their place of work. The results showed that histamine reactivity was an important, but not obligatory, factor in the development of occupational asthma, and that it appeared to be the result rather than the cause of occupational asthma, as it returned towards normal in workers removed from exposure. TDI and MDI were shown to be acting as specific causes of occupational asthma rather than nonspecific irritants at concentrations up to 0.02 ppm. There was evidence that some irritant reactions to colophony were occurring at exposure levels encountered at work, but that the majority of workers with colophony asthma were having specific reactions to the colophony fume. PMID- 6297951 TI - Behavioral and central visual correlates of inherited retinal degeneration in the domestic chicken (Gallus domesticus). AB - A developmental, posthatch loss of cutaneous and ocular melanocytes in the mutant DAM (delayed amelanotic) chicken is accompanied by severe retinal degeneration. Using a food-location task, increased ocular pigment loss was associated with increased latency of response, both developmentally in young DAMS, and among adults with differing degrees of amelanosis. Analysis of optic nerve fiber composition in normals and DAMs showed a reduction in the small diameter optic axon population in severely amelanotic DAMs. With increasing severity of ocular amelanosis among animals, reduction in the density of anterogradely transported horseradish peroxidase (HRP) reaction product was seen first in retinorecipient thalamic nuclei, subsequently in the optic tectum, and ultimately in the accessory optic nucleus. An inverse relationship was also observed between the density of HRP reaction product occurring in retinal terminal fields and density of HRP label seen in the optic tracts of the same animal. No changes in either the volume or extent of central visual nuclei were apparent in partially sighted and blind DAMs. PMID- 6297952 TI - Single thalamic neurons which project to both the rostral cortex and caudate nucleus studied with the fluorescent double labeling method. AB - After injections of fast blue into the rostral cortex and Evans blue into the caudate nucleus in cats, doubly labeled neurons were present in the ventral anterior, ventral lateral, rhomboid, and mediodorsal thalamic nuclei. Doubly labeled cells were also found in most members of the intralaminar group, including the central medial, paracentral, central lateral, and parafascicular nuclei. Although the centromedian nucleus contained large numbers of cells labeled with Evans blue which project to the caudate nucleus, and a few fast-blue labeled cells which projected to the cortex, doubly labeled neurons were absent from this posterior intralaminar nucleus in this study. PMID- 6297953 TI - Topography of locus ceruleus neurons projecting to the area dentata. AB - The distribution of locus ceruleus neurons projecting to the dorsal and ventral area dentata was studied using the retrograde transport of horseradish peroxidase (HRP). Furthermore, the possibility that collaterals of single locus ceruleus neurons innervate both the dorsal and ventral area dentata was explored with a retrograde double-label technique using HRP and the fluorescent dye 4',6 diamidino-2-phenylindole 2 HCl (DAPI). Fusiform and multipolar neurons of the pars dorsalis of the locus ceruleus (LCd) were labeled after injection of HRP into the area dentata. The HRP-positive cells were found almost exclusively in the dorsal LCd after dorsal area dentata injections whereas ventral injections labeled neurons throughout the LCd. The highest density of LCd neurons labeled from the ventral area dentata was observed in the dorsal half of the LCd and thus overlapped with the distribution of neurons labeled from dorsal injections. Neurons containing both HRP and DAPI were observed in the dorsal LCd suggesting that the topographic coincidence of neurons labeled from dorsal and ventral area dentata was in part a result of single neurons having terminals in both loci. PMID- 6297954 TI - Consequences of misrouting goldfish optic axons. AB - Cut optic axons regenerate into the goldfish tectum by indirect routes. To study the accuracy with which they terminate we made visuotectal maps before and immediately after cutting through the rostral tectum from its medial edge. This severed the normal pathway to medial tectum leaving intact only grossly misrouted axons. In fish mapped in this way, 3 months or more after contralateral optic nerve cuts, each initial map was essentially normal. After a tectal cut the electrical responses were usually weaker; but their receptive field positions proved to match very closely those noted previously for the same recording sites. The mean angular change was only 8.48 degrees and much of this could be accounted for by experimental errors. In fish mapped 6 months or more after attempts to cross-unite the optic tract brachia, initial maps were less regular. Some showed gross rearrangement of groups of terminals; and recognizable irregularities persisted in two fish mapped again after a further 4 or 5 months. However, maps made after tectal cuts in such fish were not noticeably less regular than the initial maps. Field position changes averaged 17.85 degrees. In both groups misrouted axons, filled with horseradish peroxidase after mapping and visualized in whole-mounts, crossed the tectum from lateral regions to reach the medial recording sites. The small field position changes in both groups confirm that axons from detectable terminals only among their retinal neighbours; nevertheless the irregular maps seen after tract cross-union lead us to expect the orderly arrangement of the normal pathway to contribute significantly to the precision of the normal map. PMID- 6297956 TI - Modifications of membrane cholesterol content affects electrical properties and prolactin release of cultured pituitary cells. AB - Treatment of cloned pituitary cells (GH3/B6) with cholesterol-enriched liposomes, which presumably increases membrane cholesterol content, affects the passive and active electrophysiological properties and stimulates the release of prolactin (PRL). PMID- 6297957 TI - Production of superoxide by neutrophils. AB - Oxygen uptake by neutrophils has been stimulated by particulate serum-treated zymosan (STZ) and soluble N-formylmethionyl-leucyl-phenylalanine (FMLP) in the presence and absence of the superoxide radical scavenger, cytochrome C. Results indicate that FMLP may stimulate neutrophils to produce superoxide but that STZ may not. PMID- 6297959 TI - Cyclic nucleotides phosphodiesterase activity changes in early chick development. AB - Differences in the activity of cyclic nucleotides phosphodiesterase develop in different germ layers during the gastrulation of the chick embryo. PMID- 6297958 TI - Influence of N-methylformamide on the development, the NAD synthesis, and the activity of the ADPR transferase of rat embryos. AB - When N-methylformamide is administered to rats on the 11th day of pregnancy approximately 50% of the fetuses are resorbed and a reduced weight of the developed animals is found in comparison to the controls on the 21th day (delivery by Caesarian section). The toxic effect is increased by using nicotinamide and methionine. If a combination of these substances is employed practically all fetuses are resorbed. Tryptophan, however, has a considerably protective influence. N-Methylformamide has no influence on the NAD-synthesis induced by nicotinamide or tryptophan. It does, however, inhibits the activity of the ADPR transferase. PMID- 6297960 TI - Effect of ketoconazole and miconazole on skeletal muscle mitochondrial calcium transport system. PMID- 6297961 TI - Biochemical studies of CELO virus: an oncogenic avian adenovirus. PMID- 6297962 TI - [Harman and 3-methylharman--blockaders of presynaptic alpha 2-adrenoreceptors]. AB - It has been shown in experiments on rat vas deferens that melipramine, harmane and 3-methylharman in low concentrations of the order of 10(-8)--10(-6) M increase the contraction of the vas deference induced by noradrenaline and transmural stimulation. Meanwhile in higher concentrations (10(-6)--4 X 10(-5) M) they have been discovered to decrease this effect (except for harmane). In low concentrations harmane and 3-methylharman competitively remove the inhibitory effect of clonidine on the transmural stimulation-induced contractions. It is inferred that harmane and methylharman are competitive blockers of presynaptic alpha 2-adrenoceptors in adrenergic synapses. PMID- 6297963 TI - [Effect of strophanthin in adrenergic blockade on the blood supply and work of the heart in acute coronary artery occlusion]. AB - The effects of strophathin in a dose of 0.15 mg/kg bw were investigated under alpha- and beta-adrenergic blockade by the methods of thermodilution and circulation registration in the pool of the occluded coronary artery in anesthetized dogs. It was determined that strophanthin increased the blood supply of the myocardial ischemic centres, the minute heart volume, and systemic arterial pressure and decreased the general peripheral vascular resistance. The alpha-adrenoblocker, phentolamine (0.025 mg/kg intravenously) decreased the strophanthin stimulant effect on the collateral coronary blood stream and heart activity without changing its chronotropic and vasopressive action. The beta adrenoreceptor blockade induced by obsidan (0.5 mg/kg) made no changes in the pattern of the glycoside cardiovascular effects and diminished its chronotropic action. PMID- 6297965 TI - NdeI: a restriction endonuclease from Neisseria denitrificans which cleaves DNA at 5'-CATATG-3' sequences. PMID- 6297964 TI - [Tropane ligands of different types of opiate receptors]. AB - Interaction of tropane derivatives (motropin, atropine, cocaine) with opiates (morphine) and opioids (an enkephalin amide analog) was studied according to varying tests: pain sensitivity, impulse summation in the central nervous system, respiration. It appeared that motropin is a morphine antagonist and enkephalin amide analog from the standpoint of effect on analgetic action and impulse summation, but is not their antagonist as regards the effect on respiration. Atropine is a weak morphine antagonist in terms of the effect on analgesia, impulse summation and respiration as well. Cocaine is a morphine synergist as regards all the tests indicated. Therefore, the effect of tropane derivatives on pain sensitivity, impulse summation and respiration is mediated via different opiate receptors, which does not exclude the involvement of other neurochemical mechanisms in their action. PMID- 6297966 TI - Identification of a Mr 58 000 glycoprotein subunit of the opiate receptor. AB - A Mr 58 000 subunit of the opiate receptor has been identified using tritiated fentanyl isothiocyanate, a potent opiate alkylating reagent with specificity for the delta-opiate receptor subclass. The subunit is alkylated in the presence of dextrorphan but not levorphanol. The specifically labelled protein was retained on columns of immobilized wheat germ agglutinin and is therefore presumably a glycoprotein. Partial purification of the Mr 58 000 opiate receptor subunit from neuroblastoma X glioma NG108-15 hybrid cell membranes is described. PMID- 6297955 TI - DNA polymerases in prokaryotes and eukaryotes: mode of action and biological implications. PMID- 6297967 TI - Forskolin stimulates adenylate cyclase and iodine metabolism in thyroid. AB - Forskolin is a potent activator of the cyclic AMP-generating system in many tissues. In dog thyroid slices, the enhancement of cyclic AMP level was rapid, sustained in the presence of forskolin, but easily reversible after its withdrawal. Contrary to TSH, forskolin induced little apparent desensitization. Forskolin potentiated the effects of TSH, PGE1 and cholera toxin. However, the forskolin-induced cyclic AMP accumulation was still sensitive to inhibitors of dog thyroid adenylate cyclase such as iodide, norepinephrine and adenosine. As fluoride, but contrary to TSH and PGE1, forskolin stimulated adenylate cyclase in a medium where Mg2+ was replaced by Mn2+. This suggests that in thyroid, as in other tissues, forskolin acts beyond the receptor level but, as it potentiates hormone action and does not impair modulation by inhibitors, it may interact with the nucleotide-binding regulatory proteins. Forskolin mimicked the effect of TSH on iodide organification and secretion. PMID- 6297968 TI - Activation of protein kinase isoenzymes under near physiological conditions. Evidence that both types (A and B) of cAMP binding sites are involved in the activation of protein kinase by cAMP and 8-N3-cAMP. AB - cAMP-dependent protein kinase I and II (cAKI and cAKII) were incubated under near physiological conditions in the presence of various concentrations of 8-N3 c[3H]AMP or c[3H]AMP. Both types (A and B) of cyclic nucleotide binding sites of cAKI or cAKII were occupied to a similar extent and the degree of their occupation correlated with the degree of kinase activation. cAKI and cAKII bound cAMP in an apparent positively cooperative manner in the presence of Mg2+, ATP. 8 N3-c[3H]AMP dissociated several orders of magnitude faster from site A than site B of the regulatory moiety of cAKII, and was photo-incorporated only when bound to site B. PMID- 6297969 TI - Role of Ca2+ in regulating the level of mitochondrial pyrophosphate. Effect on butyrate oxidation. PMID- 6297971 TI - Effect of ribonuclease H from chick embryo on the covalent-linked poly(A)- poly(dA) complementary to poly(dT) template. AB - In vitro poly(dA) synthesis on poly(dT) template can be initiated by poly(A) primer. Poly(A) chains are covalently extended by DNA polymerase. The reaction product consists of poly(dA) chain with poly(A) at their 5'-ends, hydrogen bonded to the template poly(dT). The primer poly(A) is linked to the product poly(dA) via a 3':5'-phosphodiester bond, and can be specifically removed by ribonuclease H from chick embryos, leaving a 5'-phosphate end of poly(dA). Poly- or oligoriboadenylate longer than the (pA)5 could serve as a priming activity to synthesize poly(A) covalently linked to poly(dA). PMID- 6297970 TI - Inactivation of spermidine N1-acetyltransferase with alkaline phosphatase. AB - Spermidine N1-acetyltransferase in an extract from phytohemagglutinin-stimulated bovine lymphocytes was inactivated by preincubation with alkaline phosphatase. Inactivation of the acetylase with the phosphatase was totally inhibited by addition of pyrophosphate. These results suggest that spermidine N1 acetyltransferase, the rate-limiting enzyme in the biodegradative pathway of polyamines, is inactivated by dephosphorylation. A similar effect of alkaline phosphatase on the acetylase in an extract from Escherichia coli was also observed. The acetylase has a rapid rate of turnover and the rapid loss of the enzyme activity may be to some extent regulated by the covalent modification. PMID- 6297972 TI - Different polypeptides of bovine heart cytochrome c oxidase are in contact with cytochrome c. AB - Two water-soluble carbodiimides, differing in molecular dimensions, have been used to characterize the cytochrome c binding site of bovine heart cytochrome c oxidase. Several polypeptide components of the enzyme contain acidic residues which are modified by these reagents. Carboxyl groups present in subunit II, VII and polypeptide c, are protected from modification when cytochrome c, equimolar to oxidase, is added and they can cross-link to the substrate once activated by the carbodiimide. Comparison of the modification patterns suggest that the most reactive residues are located on subunit II and VII, the former being also more exposed. The data obtained indicate that even though subunit II plays the major role in binding cytochrome c, at least two other lower Mr polypeptides contribute to the cytochrome c binding domain. PMID- 6297973 TI - Mechanism of lactose-proton cotransport in Escherichia coli. Kinetic results in terms of the site exposure model. AB - Rigorous kinetic derivations are presented for the Site Exposure mechanism of lactose-proton cotransport in E. coli [J. Theor. Biol. (1978) 75, 35-50]. Proton translocation inwards is solely associated with the external exposure of the galactoside binding site. A symmetric dimer configuration of the transporter is proposed, resulting in two forms corresponding to the cis and the trans orientation of the binding sites. The cis to trans orientation is inherently unfavorable, induced only by transmembrane substrate gradients. Recently reported extensive kinetic data are straightforwardly predicted by this mechanism, including the complicated effects on the apparent affinity and maximal velocity of uptake exhibited by changes in the magnitude of the proton electrochemical gradient. PMID- 6297974 TI - The reaction of cytochrome c with [Fe(EDTA)(H2O)]-. PMID- 6297975 TI - The generation of hydroxyl radicals following superoxide production by neutrophil NADPH oxidase. PMID- 6297976 TI - Carbamoylation of Cu,Zn-superoxide dismutase by cyanate. Role of lysines in the enzyme action. AB - Reaction with cyanate leads to a reversible change of the EPR spectrum of Cu,Zn superoxide dismutase and to time-dependent carbamoylation of the lysine residues of the enzyme, producing a stable covalent derivative with more negative charge. The carbamoylated enzyme is less active than the native enzyme in spite of unaltered EPR spectra. The extent of this inactivation is much less when the enzyme activity is measured at low ionic strength. These results show that integrity of the active site is not the sole factor playing a role in the enzyme mechanism and that the ionic strength effect is related to electrostatic interactions between O-2 and surface charges of the protein. PMID- 6297977 TI - The substructure of phosphodiesterase as established by radiation inactivation. A reinterpretation of results. AB - A model for the activation of phosphodiesterase by calmodulin based on a conversion of inactive dimers to active monomers, derived from radiation inactivation studies J. Biol. Chem. (1981) 256, 11351-11355 has been re-examined using a simple probability argument. We conclude that the original model is not supported by the radiation inactivation studies, since our analysis of this model would predict that the rate of radiation inactivation of calmodulin-dependent phosphodiesterase activity be exactly twice that for the decay in total activity in marked contrast with the results obtained. PMID- 6297978 TI - Reconstitution of a Mg-ATP-dependent protein phosphatase and its activation through a phosphorylation mechanism. AB - A Mg-ATP-dependent protein phosphatase has been reconstituted from the catalytic subunit of protein phosphatase-1 and inhibitor-2, and consists of a 1:1 complex between these proteins. Activation of this enzyme by glycogen synthase kinase-3 and Mg-ATP results from the phosphorylation of inhibitor-2 on a threonine residue(s) and is accompanied by the dissociation of the complex. The results prove that protein phosphatase-1 and the Mg-ATP-dependent protein phosphatase contain the same catalytic subunit, and that they are interconvertible forms of the same enzyme. PMID- 6297979 TI - Does a cyclic AMP-dependent phosphorylation initiate the transfer of trehalase from the cytosol into the vacuoles in Saccharomyces cerevisiae? AB - Trehalase activity in a yeast protoplast lysate increased approximately 40-times upon preincubation with cAMP and ATP. The activity present without the preincubation could all be sedimented at 8000 x g, for 10 min confirming the previously reported localization of the active trehalase (Ta) in the vacuoles. Virtually all the trehalase activity newly formed upon the preincubation, however, was found in the soluble fraction, indicating that a trehalase-zymogen (Tz) is located in the cytosol. This raises the possibility that a cAMP-dependent phosphorylation not only transforms Tz to Ta but also initiates the transfer of trehalase from the cytosol into the vacuoles. PMID- 6297980 TI - The glucagon receptor from liver can be functionally fused to caudate nucleus adenylate cyclase. PMID- 6297981 TI - The role of superoxide and hydroxyl radicals in phospholipid peroxidation catalysed by iron salts. AB - Iron(II) salts in aqueous solution, or iron(III) salts in the presence of an O.-2 generating system, can activate dioxygen to produce hydroxyl radicals. These are detected indirectly by their ability to degrade deoxyribose with the formation of thiobarbituric acid-reactive (TBA) products. Iron salts also catalyse the peroxidation of phospholipids resulting in the formation of TBA-reactive products. Hydroxyl radicals were responsible for the degradation of deoxyribose but not for the observed peroxidation of phospholipid. The function of O.-2 in both deoxyribose degradation and phospholipid peroxidation seems to be that of reducing iron(III) into iron(II). PMID- 6297982 TI - On the mechanism of biosynthesis of leukotrienes and related compounds. AB - [10D-3H; 3-14C]- and [10L-3H; 3-14C]arachidonic acids were incubated with human polymorphonuclear leukocytes and with human platelets. Leukotriene B4 and 5(S),12(S)-dihydroxy-6trans,8cis,10trans,14-cis-eicosatetraenoic acid (5,12 DHETE) were isolated and the 3H/14C ratios determined. It could be concluded that the 10D (pro-R)-hydrogen is eliminated in the conversion of 5(S)-hydroperoxy 6trans,8-cis,11cis,14cis-eicosatetraenoic acid into leukotriene A4 whereas in the conversion of arachidonic acid into 5,12-DHETE the 10L (pro-S)-hydrogen is lost. Incubation of the doubly labeled arachidonic acids with human platelets confirmed and extended previous data on the stereochemistry of the hydrogen removal from C 10 during the conversion into 12(S)-hydroperoxy-5cis,8cis,10trans,14cis eicosatetraenoic acid, i.e., the 10L (pro-S)-hydrogen is eliminated and the 10D (pro-R)-hydrogen retained. PMID- 6297983 TI - Solubilization and partial purification of the high affinity [3H] imipramine binding site from human platelets. PMID- 6297984 TI - Dependence on pH of substrate binding to lactose carrier in Escherichia coli cytoplasmic membranes. AB - Lactose permease in Escherichia mediates proton-substrate cotransport. The molecular mechanism of this process is not understood. We examined the effect of proton concentration on the binding of a substance analogue to the carrier. The dissociation constant of p-nitrophenyl-alpha-galactoside from the carrier was dependent on pH, with an apparent pKa of 9.7. PMID- 6297985 TI - Detection of Sendai virus fusion with human erythrocytes by fluorescence photobleaching recovery. PMID- 6297986 TI - Decrease in uH2A (protein A24) of a mouse temperature-sensitive mutant. AB - ts85 cell is a temperature-sensitive mutant of cell cycle, and chromosomal protein uH2A of this mutant disappears at the non-permissive temperature. uH2A in nucleosomes is thought to be synthesized or degradated as follows. H2A + Ubiquitin in equilibrium uH2A. Up to date, the degradation of uH2A was shown to be catalyzed by uH2A lyase, however no enzymes (factors) concerning its synthesis have been elucidated. Here, we show that ATP is prerequisite for the synthesis of uH2A, and that the disappearance of uH2A at the non-permissive temperature may be due to a reduction in the rate of synthesis rather than an increase in the rate of its degradation. PMID- 6297987 TI - Induction of maturation in rat follicle-enclosed oocyte by forskolin. AB - The diterpene forskolin, which was found to be a potent and reversible activator of adenylate cyclase in intact tissues as well as in broken cell preparations, was employed to investigate the role of cAMP in the induction of oocyte maturation. We have found that forskolin can mimic the effect of LH on the ovarian follicle stimulating both cAMP accumulation and oocyte maturation. These findings suggest that LH-induced maturation in follicle-enclosed oocytes is a cAMP-mediated response. PMID- 6297988 TI - DNA sequence of the promoter region of the ompC gene and the amino acid sequence of the signal peptide of pro-OmpC protein of Escherichia coli. AB - The osmoregulated ompC gene of Escherichia coli was cloned and the DNA sequence of a fragment encompassing the promoter region and a portion of the coding region was determined. There were no obvious homologies in the DNA sequences of the promoter regions of the ompC and ompF genes, in contrast to those of the coding regions of the two genes, both of which code for the matrix porins (major outer membrane proteins) and form passive diffusion pores. The amino acid sequence of the signal peptide of pro-OmpC protein was also deduced from the DNA sequence. PMID- 6297989 TI - Cyclic AMP-independent phosphorylation of Escherichia coli isocitrate dehydrogenase. AB - The phosphorylation of NADP-specific isocitrate dehydrogenase in a wild-type and in an adenylate cyclase deletion mutant of Escherichia coli has been investigated. The results obtained clearly indicate that cyclic AMP is not required for the phosphorylation reaction per se, not is it for the synthesis or possible activation of the phosphoprotein kinase in this organism. This data are in contrast to results observed in Salmonella typhimurium, and indicate that important differences exist in the phosphorylation of the isocitrate dehydrogenase in these two organisms. PMID- 6297991 TI - Hydrophobic properties of Tetrahymena calmodulin related to the phosphodiesterase activity. AB - We have examined hydrophobic properties of Tetrahymena CaM using the uncharged probe, n-phenyl-1-naphthylamine (NPN) fluorescence. The maximal fluorescence intensity of Tetrahymena calmodulin (CaM) is less than 1/12 of that of the bovine brain CaM. In the phosphodiesterase activation, the potency of Tetrahymena CaM, which was represented by reciprocals of the quantity of CaM required for half maximal activation of enzyme was 22.7% respectively, of that of the bovine brain CaM. Here, Tetrahymena CaM had less hydrophobic groups exposed in the presence of Ca2+. Then Ca2+-CaM dependent enzymes require much amount of Tetrahymena CaM, comparing with the bovine brain CaM. PMID- 6297990 TI - The CCK receptor on pancreatic plasma membranes: binding characteristics and covalent cross-linking. AB - The cholecystokinin (CCK) receptor in purified plasma membranes prepared from mouse pancreatic acini had a binding affinity of 1.8 nM, an acid pH optimum between 6.0 and 6.5, and an analog specificity of CCK8 greater than CCK33 greater than desulphated CCK8 greater than CCK4. Binding of CCK to its receptor was abolished by pretreatment of plasma membranes with trypsin. When [125I]CCK was cross-linked to its receptors with disuccinimidyl suberate, and the preparation solubilized and subjected to gel electrophoresis and autoradiography, the hormone was associated with Mr 80 000 protein in both the presence and absence of the reducing agent dithiothreitol. PMID- 6297992 TI - Cyclic AMP-dependent protein kinase does not phosphorylate cyclic GMP-dependent protein kinase in vitro. AB - The autophosphorylation reaction of purified cGMP-dependent protein kinase has been studied. Apparent initial rates of autophosphorylation in the absence of cyclic nucleotides and in the presence of cGMP and cAMP are 0.006, 0.04, 0.4 mol Pi incorp./min-1. mol cGMP-kinase subunit-1. In the presence of cGMP and cAMP approximately 1 and 2 mol Pi are incorporated/mol enzyme subunit. These values are independent of the enzyme concentration. Stimulation of autophosphorylation by cAMP is not due to activation of a contaminating cAMP-dependent protein kinase since: (a) addition of the heatstable inhibitor protein of cAMP-kinase does not inhibit autophosphorylation; and (b) catalytic subunit of cAMP-kinase added at a 10-fold excess over cGMP-kinase does not phosphorylate cGMP-kinase. PMID- 6297993 TI - Tryptophanyl-tRNA synthetase: pyrophosphorylation of the enzyme in the course of adenylate formation? PMID- 6297994 TI - Parathyroid hormone secretion in the absence of extracellular free Ca2+ and transmembrane Ca2+ influx. AB - The involvement of extracellular Ca2+ and Ca2+ influx across the plasma membrane in parathyroid hormone (PTH) secretion was investigated in vitro using a new preparation of bovine parathyroid cells. Incubation of these cells in the presence of 25 microM or 2.5 microM free ambient Ca2+ induced a maximal rate of PTH secretion. Low free Ca2+ secretion is not associated with changes in membrane permeability, requires metabolic energy, and is reversible. The Ca2+ channel blocker D600 had no effect on either 45Ca-influx or PTH secretion in these cells. These results, showing that extracellular Ca2+ and Ca2+ influx across the plasma membrane are not required for PTH secretion by parathyroid cells, emphasize the differences in the cellular mechanisms underlying the secretion of PTH vs that of other secretory cells. PMID- 6297995 TI - Vinblastine-induced autophagocytosis: effects on liver glycogen. AB - The possible similarities of the mechanism by which vinblastine induces autophagocytosis in liver were compared with the known effects of glucagon in glucagon-induced autophagocytosis. A single intraperitoneal injection of vinblastine produced a wave of autophagocytosis in less than 0.5 h in mouse hepatocytes. Liver glycogen content decreases simultaneously and blood glucose first increased and then decreased below control values. Both liver cAMP concentration and the activity of glycogen phosphorylase remained unchanged. These findings provide evidence that the induction of autophagocytosis after vinblastine injection is not mediated by cAMP. The increased degradation of glycogen may occur in the lysosomal system by means of increased autophagocytosis. PMID- 6297996 TI - On the presence of two new high mobility group-like proteins in HeLa S3 cells. AB - Two phosphorylated HMG-like proteins with Mr approximately 10 000 have been isolated from HeLa S3 cells, one being present in metaphase and one in interphase cells. The amino acid compositions of these proteins are very similar but differ from the known HMG proteins. However, they exhibit similarities being rich in proline, basic and acidic amino acids. A possible role in chromatin condensation of the HMG-like protein characteristic for metaphase cells is suggested. PMID- 6297997 TI - Cyclic AMP-dependent protein kinase stimulates the formation of polyphosphoinositides in lymphocyte plasma membrane. AB - Inside-out vesicles from lymphocyte plasma membrane were phosphorylated in the presence of [gamma -32P]ATP. The dissociated catalytic subunit of cyclic AMP dependent protein kinase stimulated both membrane protein and membrane lipid phosphorylation, indicating the presence of a phosphorylation cascade. The phosphorylated membrane lipids were analyzed by thin-layer chromatography. Increase of 32P-labelling stimulated by the cyclic AMP-dependent protein kinase was found exclusively in polyphosphoinositides. PMID- 6297998 TI - Differences in the association between the oxidase-dependent activity and plasma membrane receptors for IgG, C3b and concanavalin A of human neutrophils. PMID- 6297999 TI - Decreased activity of uroporphyrinogen decarboxylase caused by 2,4,5,3',4' pentabromobiphenyl in chick embryo hepatocyte cultures. Difference in activity in intact or homogenized cells. AB - Uroporphyrinogen decarboxylase activity was investigated in cultures of chick embryo liver by two different methods: (1) analysis of porphyrin composition following incubation of intact cells with delta-aminolevulinic acid; and (2) a more conventional direct enzymic assay of cell homogenates. Activity was detectibly decreased following exposure of cells to 100 ng/ml 2,4,5,3',4' pentabromobiphenyl using the first method, but not the second. This decrease in activity was reversed by homogenizing the cells treated with 100 ng/ml pentabromobiphenyl. It is concluded that the direct homogenate assay of the enzyme may miss or underestimate decreases in its in vivo activity. PMID- 6298000 TI - Characterization of a new membrane-bound cytochrome c of Rhodopseudomonas capsulata. AB - A cytochrome c (cyt. c) was solubilized with Triton-X-100 and co-purified with cytochrome c oxidase from membranes of chemotrophically grown cells of Rhodopseudomonas capsulata. Cyt. c and cytochrome oxidase were separated on Sephadex G-50 columns. Antibodies against cytochrome c2 from the same bacterium did not cross react with the membrane-bound cyt. c. The IEP of the membrane-bound cyt. c was found to be pH 8.2, the midpoint potential was 234 +/- 11 mV at pH 7.0. This cyt. c binds CO. The native cyt. c is a dimer with an apparent Mr of 25000 containing 2 mol heme per mol dimer, which is believed to function as an electron donor for the high-potential cytochrome c oxidase. PMID- 6298001 TI - High affinity binding of [125I]monoiodoapamin to isolated guinea-pig hepatocytes. AB - The bee venom neurotoxin apamin has been labelled with 125I and its binding to isolated guinea-pig hepatocytes measured under physiological conditions. A single saturable component of [125I]monoiodoapamin binding with a Kd of 350 pM and Bmax of 0.99 fmol/mg dry wt was identified. Native apamin displaced labelled apamin with a Kd of 376 pM which is broadly in keeping with the concentrations found to inhibit K loss from guinea-pig hepatocytes. These observations, together with the binding found in other tissues, suggest that specific binding of labelled apamin is particularly associated with apamin-sensitive, Ca-activated K-channels. PMID- 6298002 TI - Calcium inhibition of a heat-stable cyclic nucleotide phosphodiesterase from Neurospora crassa. AB - Neurospora crassa had a heat-stable (up to 95 degrees C), soluble cyclic nucleotide phosphodiesterase (PDE). Both unheated and heat-stable PDE activities were inhibited by micromolar concentrations of Ca2+. This inhibition was reversed by EGTA or EDTA in molar excess of the Ca2+ concentration. Calmodulin was not involved in the Ca2+ inhibition, nor was Ca2+ inhibition of the heat-stable PDE due to cleavage inactivation of the enzyme by a Ca2+-stimulated protease. In addition to Ca2+, several other cations inhibited the activity of the heat-stable enzyme. Cyclic AMP and cGMP, but not 2'3' cAMP were substrates for both unheated and heat-stable PDEs. This is the first report of a PDE which is inhibited by micromolar concentrations of Ca2+. PMID- 6298003 TI - Characterization and synthesis of a macrophage inhibitory peptide from the second constant domain of human immunoglobulin G. AB - We have shown that IgG hydrolysed by Schistosoma mansoni schistosomula inhibited various macrophage functions, especially phagocytosis and anti-schistosome cytotoxicity. Here we show that a tripeptide, Thr289-Lys-Pro291, of the second constant domain of human immunoglobulin G (peptide 286-292) reproduced the inhibitory effect of a total hydrolysate. Indeed the beta-glucuronidase release from IgE-anti-IgE-stimulated rat and human macrophages decreased and its intracellular level did not rise after a prior incubation of the cells with Thr Lys-Pro (500 nmol/ml). Moreover, the cell migration as well as the superoxide anion O2 generation were 50-80% reduced by the tripeptide. These results suggest that a single peptide set may be responsible for the decrease of the macrophage functions at the early stage of the parasite infection in the mammalian host. The pharmacologic properties of this tripeptide are under investigation. PMID- 6298004 TI - Vitamin D stimulates (Na+ + K+)-ATPase activity in chick small intestine. AB - Vitamin D3 and 1,25-dihydroxyvitamin D3 raise (Na+ + K+)-ATPase activity (ouabain sensitive 86Rb+ uptake) in cultured embryonic and 4-week-old chick small intestine. Vitamin D stimulation of the sodium pump, which requires genomic action of the sterol, may lead to enhanced Ca2+ extrusion via a basolateral Na+/Ca2+ exchange mechanism, and, in addition, may provide a proliferative signal in undifferentiated enterocytes. PMID- 6298005 TI - Characterization of cytochrome b in the isolated ubiquinol-cytochrome c2 oxidoreductase from Rhodopseudomonas sphaeroides GA. AB - Extinction coefficients for cytochrome b and c1 in the isolated cytochrome bc1 complex from Rhodopseudomonas sphaeroides GA have been determined. They are 25 mM 1 . cm-1 at 561 nm for cytochrome b and 17.4 mM-1 . cm-1 at 553 nM for cytochrome c1, for the difference between the reduced and the oxidized state. Cytochrome b is present in two forms in the complex. One form has an Em7 of 50 mV, an alpha peak of 557 nm at liquid N2 temperature and of 561 nm at RT, which is red-shifted by antimycin A. The other form has an Em7 of -90 mV, a double alpha-peak of 555 and 561 nm at liquid N2 temperature corresponding to 559 and 566 nm at RT. The absorption at 566 nm is red-shifted by myxothiazol. The two shifts are independent of each other. Both midpoint potentials of cytochromes b are pH dependent. The redox center compositions of the cytochrome bc1 complexes from Rhodopseudomonas sphaeroides and from mitochondria are identical. PMID- 6298006 TI - Cobra polypeptide cytotoxin I and marine worm polypeptide cytotoxin A-IV are potent and selective inhibitors of phospholipid-sensitive Ca2+-dependent protein kinase. AB - The effects of a number of polypeptide cytotoxins and neurotoxins on various protein kinases were examined. It was found that cobra cytotoxin I and marine worm cytotoxin A-IV effectively and specifically inhibited phospholipid-sensitive Ca2+-dependent protein kinase (PL-Ca-PK) relative to myosin light chain kinase and cyclic nucleotide-dependent protein kinases. Inhibition of PL-Ca-PK by these cytotoxins could be overcome by phosphatidylserine. Neurotoxins, in comparison, were much less effective inhibitors. The present findings indicated that these polypeptide cytotoxins, unlike other agents reported to date, were selective inhibitors of PL-Ca-PK and could be used to differentiate Ca2+-dependent events regulated by phospholipid or calmodulin. PMID- 6298007 TI - Forskolin as a tool to study the beta-adrenergic receptor-elicited, labeled protein secretion in rat lacrimal gland. AB - In rat lacrimal glands, Forskolin induces a dose-dependent [3H]protein release. This effect can be potentiated by papaverine. As for the other inducers whose effects on protein secretion are assumed to be cAMP-mediated, Forskolin secretion time course shows a latency. Isoproterenol decreases the Forskolin EC50 at least 60-times. On the other hand, Forskolin enhances the efficacy of isoproterenol without affecting its potency. As a whole, the data collected show that isoproterenol-induced [3H]protein secretion in rat lacrimal glands involved adenylate cyclase activation by coupling with beta-adrenergic receptors. PMID- 6298008 TI - Mitochondrial NADH dehydrogenase-catalyzed oxygen radical production by adriamycin, and the relative inactivity of 5-iminodaunorubicin. PMID- 6298009 TI - Phosphorylation of the ribosomal protein S6 in response to secretagogues in the guinea pig exocrine pancreas, parotid and lacrimal gland. PMID- 6298010 TI - Angiotensin II inhibits hepatic cAMP accumulation induced by glucagon and epinephrine and their metabolic effects. AB - Incubation of isolated hepatocytes containing normal Ca2+ levels with angiotensin II, vasopressin or A23187 caused significant inhibition of the cAMP response to glucagon. Angiotensin II also inhibited cAMP accumulation induced by either glucagon or epinephrine in Ca2+-depleted hepatocytes. When submaximal doses of hormone were employed such that cell cAMP was elevated only 3-4-fold (approximately 2 pmol cAMP/mg wet wt cells) inhibition by angiotensin II was correlated with a decrease in phosphorylase activation. The data demonstrate that inhibition of hepatic cAMP accumulation results in reduced metabolic responses to glucagon and epinephrine and do not support the contention that the hepatic actions of glucagon are independent of cAMP. PMID- 6298011 TI - Specific epidermal growth factor receptors on porcine thyroid cell membranes. AB - Specific saturable receptors for epidermal growth factor (EGF) of high affinity (Ka 1.7 X 10(9) M-1) have been demonstrated on porcine thyroid membranes. Optimal conditions for EGF binding have been determined. TSH and other peptide hormones do not inhibit the binding of 125I-EGF and EGF does not inhibit 125I-TSH binding to thyroid membranes. The results suggest that EGF may be involved in the regulation of thyroid follicular cell growth and function. PMID- 6298012 TI - [Effect of functional changes in the dopamine system of the brain on enkephalinamide-induced catalepsy in the rat]. PMID- 6298013 TI - [Functional characteristics of pairs of adjacent neurons in the rabbit inferior colliculus]. PMID- 6298014 TI - [Changes in the spatial distribution of the neuronal activity in the visual cortex after stimulation with flashes of diffuse light]. AB - At different stages of development of the response to flash, grouping of excited neurons of the guinea pig 17th area occurs within the same cortical microzones separated by narrow inhibition zones common to the neighbouring excited ones. The ensembles of excited neurons have the form of columns narrowing into depth. The grouping of cells into a silent gap indicates the grouping of inhibiting neurons. Three subgroups are differentiated by the character of averaged dynamics of impulse activity in ensembles, the essential moment of their interrelationship being the reciprocity. The one subgroup in deep layers is characterized by stability of the focus at different stages of the response development and seems to express the grouping of corticofugal neurons. The ensembles are considered to be one of the forms of activity of the structured morphofunctional cortical units, i.e. the columns. PMID- 6298015 TI - [Effects of colchicine, cyclic nucleotides and biogenic amines on the chemoreceptor apparatus of the tongue]. AB - Colchycine considerably decreased sensitivity of the Rana temporaria tongue's chemoreceptors to adequate stimulation. The effect is reversible and can be abolished by administration of drugs initiating setting of the cells' microtubular apparatus: 3',5'-cAMP, theophylline, adrenaline as well as by activation of the sympathetic system. The restoring effect of these agents is specific since 2',3'-cAMP, 5'-AMP, serotonin, inosine, acetylcholine and 3,'5' cGMP prove ineffective in restoring the reactions after colchycine administration although they do alter the activity of the receptors in normal conditions. The revealed colchycine-sensitive, 3',5'-cAMP-dependent process seems to be associated with the microtubular apparatus of the receptor cell. Its role in the receptor structures is discussed. PMID- 6298017 TI - [Preliminary studies and application of monoclonal antibodies]. PMID- 6298016 TI - In vitro cleavage of gag-myc fused protein P110 of a defective leukaemia virus MC29 by retroviral protease p15. AB - Gag-related proteins were precipitated from 35S-methionine-labelled cells of the PR-2 line derived from a hepatoma induced by virus MC29. Immunoprecipitates were incubated with viral protease p15 and the cleavage products of P110 were analysed in SDS-PAGE. The major cleavage fragment of P110 is a protein with mol. wt. of 56 000. Preparative isolation of the non-gag fragments from protein P110 by means of a gag immunosorbent showed that the 56K fragment did not contain any gag specific antigenic determinants. This finding was also confirmed by performing immunization with this fragment. Cleavage of protein P110 and the properties of cleavage products are discussed. PMID- 6298018 TI - [Progress in the area of porcelain teeth]. PMID- 6298019 TI - [Decreased rate of mitosis in human lymphocytes following contact with thymus proteins]. PMID- 6298020 TI - Subcellular responses of hepatocytes to diabetes in control-fed rats. AB - The biochemistry and ultrastructure of hepatocytes from streptozotocin-diabetic rats adapted to a controlled feeding schedule are described. The microsomal enzyme glucose-6-phosphatase (G-6-Pase), required for glucose release from the hepatocyte was monitored in homogenate preparations at times after the initiation of feeding in rats trained to a 6 h feeding, 18 h fasting cycle. G-6-Pase specific activity which is increased in ad lib fed diabetic rats was not further increased with time after the initiation of feeding in the feeding trained animals. However, the known elevation in G-6-Pase latent activity of the diabetic rat was reduced during the feeding cycle of times of minimum and maximum plasma glucose. Enzyme latency is a reflection of the multicomponent nature of G-6-Pase activity; therefore, plasma glucose levels may influence elements of that multicomponent system. Hepatic rough and smooth endoplasmic reticulum (RER + SER) fractions from the diabetic animals exhibited high and equivalent G-6-Pase specific activities independent of feeding or fasting. Ultrastructural observations of periportal hepatocytes showed a high content of SER correlated with the high G-6-Pase specific activity and closely associated with dispersed particles of glycogen at all times after the initiation of feeding. Also, an increase in SER was observed in the fasted normal animals although particulate glycogen was nearly absent. These findings support earlier work indicating that diabetes stimulates the proliferation of hepatic SER and that the membranes of this organelle are altered from those of the normal animal. PMID- 6298021 TI - Na+/K+ ATPase and cell growth: effect of epidermal growth factor on the enzymatic activity in chick embryo epidermis during the embryonal development. AB - 1. The behaviour of ATPase activity during embryonic development of chick embryo epidermis has been studied in the absence or presence of a single inoculation of EGF at the fifth day from fertilization (0-day). 2. EGF strongly decreases ATPase activity by affecting Na+/K+ ATPase. This effect occurs only if begun at 0-day. 3. This effect is due to the EGF induced decrease of -SH groups that are active part of Na+/K+ ATPase. PMID- 6298022 TI - Rat brain (Na+-K+)ATPase: modulation of its ouabain-sensitive K+-PNPPase activity by thimerosal. AB - 1. The (Na+ + K+) ATPase activity of a rat brain synaptic membrane preparation was inhibited by 10(-5) M thimerosal. 2. The ouabain inhibitable K+-PNPPase activity of thimerosal treated membranes was compared with that of untreated membranes with respect to sensitivity to temperature, ouabain, K+ and ATP. 3. All those kinetic characteristics were substantially altered by treatment with thimerosal. PMID- 6298023 TI - Proteins involved in the activation and control of the two pathways of human complement. PMID- 6298024 TI - Mechanisms of catalyses: opposing relationship with enzyme stability. PMID- 6298025 TI - Control of mitochondrial respiration. PMID- 6298026 TI - The role of interaction between opiate receptors and alpha-adrenergic receptors in tolerance and dependence. PMID- 6298027 TI - Adaptation of a neuron to normorphine and clonidine. PMID- 6298028 TI - Regulation of cytoplasmic pH (pH1) in bacteria and its relationship to metabolism. PMID- 6298029 TI - Metabolic control by pump slippage and proton leakage in 'delocalized' and more localized chemiosmotic energy-coupling schemes. PMID- 6298030 TI - Stimulation by forskolin of the thyroid adenylate cyclase, cyclic AMP accumulation and iodine metabolism. AB - Forskolin, a diterpene hypotensive drug, activates adenylate cyclase in brain and in some other tissues (Seamon et al., 1981). Forskolin activated adenylate cyclase in particulate preparations and enhanced cyclic AMP accumulation in slices of dog thyroid. These effects were maximal within minutes and remained constant afterwards. The action of forskolin on intact cells disappeared rapidly after washing. It reproduced two known cyclic AMP-mediated TSH effects: the activation of secretion and of protein iodination. Forskolin thus provides a very convenient tool for the study of the action of defined elevations of cyclic AMP level in thyroid cells. The activation by forskolin of adenylate cyclase was not reduced by Mn2+ which uncouples TSH and PGE1 action. This suggests that in the thyroid also, forskolin acts beyond the receptor level. The effect of forskolin on cyclic AMP accumulation was inhibited by the known negative regulators of this system in the thyroid, acetylcholine, iodide, norepinephrine, PGF1 alpha and adenosine. On the other hand, forskolin potentiated the effects of TSH, PGE1 and cholera toxin. These data show that, though it does not require the receptors for its action, forskolin does not uncouple them from the catalytic unit of adenylate cyclase. PMID- 6298031 TI - Proteoglycan production by bovine granulosa cells in vitro occurs in response to fsh. AB - Bovine granulosa cells from small (1-9 mm) or large (10-20 mm) follicles were incubated in a chemically defined medium containing 5 muCi/ml [3H]glucosamine, gonadotropins or polypeptide hormones, 8-Br-cAMP, theophylline or trifluoperazine (TFP). Radiolabeled proteoglycans were precipitated with 10% phosphotungstic acid. Maximum incorporation of isotope occurred in 45-60 min. Radiolabeled products were completely hydrolyzed with chondroitinase ABC. FSH, but not LH or hCG, yielded a significant log-dose response. High doses of hCG inhibited the ability of granulosa to respond to FSH. No other hormone altered the FSH dose response curve. Addition of 8-Br-cAMP or theophylline mimicked the FSH response. The FSH effect was blocked by TFP, an inhibitor of calmodulin, but cAMP or theophylline overcame the effect of TEP. No distinct difference in response to these various compounds was noted between granulosa from small or large follicles. This system provides a biochemical marker for evaluating a mechanism of action for FSH. PMID- 6298032 TI - Follicle-stimulating hormone regulates in vivo testicular phosphodiesterase. AB - The effect of FSH on testicular phosphodiesterase was studied in immature rats in order to verify that the regulation of response to hormone in the gonad involves an increased cyclic AMP catabolism. Hydrolysis of cyclic AMP and cyclic GMP was measured in the homogenates of seminiferous tubules and interstitium of control animals and animals injected i.p. with 50 micrograms ovine FSH twice, 24 h and 12 h before necroscopy. After hormonal treatment, cyclic AMP phosphodiesterase activity in the seminiferous tubules was markedly increased whether results were expressed per testis or per mg protein, while cyclic GMP phosphodiesterase present in the same compartment was apparently unaffected. In the interstitium, cyclic nucleotide hydrolysis was usually decreased after FSH injection. The stimulation of phosphodiesterase, a slow process reaching maximal stimulation after 12 h, was dependent on the dose of FSH injected. In addition, DEAE cellulose chromatography of cytosol prepared from control and treated seminiferous tubules confirmed that stimulation was restricted to a cyclic AMP hydrolysing enzyme while the activity of the cyclic GMP hydrolysing form was not modified. Thus it is demonstrated that testicular phosphodiesterase is under FSH control. It is proposed that this in vivo regulation is a relevant phenomenon in the modulation of Sertoli cell function and contributes to the refractoriness that follows gonadotropin treatment. PMID- 6298033 TI - The effect of folic acid on cAMP-elicited cAMP production in Dictyostelium discoideum. AB - In Dictyostelium discoideum, folic acid serves as a chemoattractant, accelerates the appearance of several developmentally regulated enzymes, alters the phase of spontaneous light-scattering oscillations, and elicits increases in cGMP and cAMP. The effect of folic acid and pteridine analogs on rates of [3H]cAMP secretion, monitored by rapid perfusion, was studied. At low cell densities folic acid elicited only minimal [3H]cAMP secretion. At high cell densities, folic acid elicited greater increases in cellular cAMP and rates of [3H]cAMP secretion than did maximally effective doses of cAMP. Following pretreatment with 10(-8) M cAMP for 10 min or 10(-6) M cAMP for 2 min, no [3H]cAMP was secreted in response to folic acid. Conversely, pretreatment with folic acid did not impair subsequent responsiveness to 5 X 10(-9) M cAMP. Simultaneous addition of folic acid and cAMP increased the release of [3H]cAMP two- to threefold compared to control stimuli of the same concentration of cAMP. Pretreatment with a mixed stimulus attenuated the response to a subsequent increment in the cAMP concentration only if the higher cAMP stimulus did not include folic acid. The effects of folic acid are mimicked by the pteridine ring alone; analogs substituted at the 2- or 5-position are ineffective; non-reducible analogs are partially active. The observations presented here suggest significant differences in the mechanisms by which cAMP and folic acid stimulate cAMP production. Rather than there being separate receptors for each chemoattractant that interact in the same way with an intracellular pathway, it appears that folic acid and pteridine analogs potentiate the response to cAMP. PMID- 6298034 TI - Prolyl hydroxylase activity during newt forelimb regeneration. AB - Prolyl hydroxylase activity was measured to obtain insight into changes in collagen metabolism during forelimb regeneration in the adult newt, Notophthalmus viridescens. Activity increased markedly during the redifferentiative stages, remained elevated during digit formation, and decreased as regeneration neared completion. The greatest activity, seen during early digit formation, represents a 20-fold increase in activity over the level seen in nonregenerating limbs. The profile of enzyme activity correlates with the rate of collagen synthesis and the soluble collagen content of the regenerating tissue. Enzyme extracted from limbs in the early stages of regeneration can be activated in vitro upon preincubation with cofactors; however, preincubation has little, if any, effect on enzyme extracted from limbs at the later digit-forming stages. PMID- 6298035 TI - A new class of rapidly developing mutants in Dictyostelium discoideum: implications for cyclic AMP metabolism and cell differentiation. AB - Rapidly developing (rde) mutants of Dictyostelium discoideum, in which cells precociously differentiated into stalk and spore cells without normal morphogenesis, were investigated genetically and biochemically. Genetic complementation tests demonstrated that the 16 rde mutants isolated could be classified into at least two groups (groups A and C) and that the first described rde mutant FR17 (D. R. Sonneborn, G. J. White, and M. Sussman, 1963, Dev. Biol. 7, 79-93) belongs to group A. Morphological studies revealed several differences in development and final morphology between group A and group C mutants. In group A mutants, the time required for cell differentiation from vegetative cells to aggregation competent cells is reduced, whereas the time required for spore and stalk cell differentiation following the completion of aggregation is shortened in group C mutants. This suggests that group C mutants represent a new class of rde mutants and that there exist at least two mechanisms involved in regulating the timing of development in D. discoideum. Measurements of cell-associated and extracellular phosphodiesterase activities, and intracellular and total cAMP levels revealed that cAMP metabolism in both groups is significantly altered during development. Group A mutants showed precocious and excessive production of phosphodiesterase and cAMP during the entire course of development; intracellular cAMP levels in group C mutants were extremely low, and spore and stalk cell differentiation occurred without an apparent increase in these levels. Thus, while cAMP metabolism is abnormal in all the rde mutants studied, there exist several distinct types of derangement, not necessarily involving the overproduction of cAMP. PMID- 6298036 TI - The effects of 20-hydroxyecdysone on the metabolic labeling of membrane proteins in Drosophila imaginal discs. AB - 20-Hydroxyecdysone induces evagination of imaginal discs of Drosophila melanogaster cultured in vitro. The possible involvement of cell-surface proteins in this process has prompted us to study the synthesis of membrane proteins in imaginal discs. A procedure is reported for the isolation of membrane vesicle fractions from discs that are enriched for the plasma membrane enzyme, Na+/K+ ATPase, and that label with the surface-labeling reagent [125I]iodosulfanilic acid. 20-Hydroxyecdysone alters the pattern of [35S]methionine incorporation into polypeptides in these membrane vesicle fractions. Increased and decreased incorporation as well as changes in migration on two-dimensional gels of specific polypeptides are caused by the hormone. These changes parallel in time the onset and the continuation of evagination. PMID- 6298037 TI - Odor-aversion learning by rats following LiCl exposure: ontogenetic influences. AB - The influence of LiCl preexposure on odor-aversion conditioning produced by an odor-LiCl pairing was studied in rats 7-31 days old. LiCl preexposure interfered with the acquisition of the odor aversion in both young and old pups. The time course of this effect depended, however, on the age of the pups. It lasted much longer (over 24 hr) if pups 8-17 days old were preexposed to LiCl than if the pups were preexposed when 30 days old (less 12 hr). Alternative interpretations of these data were discussed, and it was suggested that the age-dependent differences in the time course of the LiCl preexposed effect may be determined in part by age-related differences in the rat's excretory processes that clear lithium from the circulatory system. PMID- 6298038 TI - Central and peripheral nervous system complications. AB - Symptomatic neuropathy is a common manifestation of diabetes mellitus, and sensory, motor, or autonomic symptoms occur in approximately 10% of all diabetic patients. Animal models may be useful to study the metabolic and electrophysiologic abnormalities peculiar to diabetic neuropathy. Genetic animal models, including the Chinese hamster, ob/ob mouse, db/db mouse, BB-wistar rat, and SSDR rat or chemically induced or nutritional models of diabetes mellitus provide the potential to use animals to study human neuropathy; however, to date, few characteristics of human diabetic neuropathy have been clearly demonstrated in any of these animal models. Better characterization of the neuropathy of existing animal models with emphasis on evaluation over long periods of time is recommended. These studies should include a cross-disciplinary approach using biochemical, electrophysiologic and morphologic techniques. Specific future approaches to study diabetic neuropathy using chemical models is outlined in this chapter. PMID- 6298039 TI - Important role of adrenergic mechanisms in acute glucose counterregulation following insulin-induced hypoglycemia in type I diabetes. Evidence for an effect mediated by beta-adrenoreceptors. AB - During hypoglycemia induced by an i.v. insulin infusion for 60 min, rates of plasma glucose (PG) decrease and recovery, PG nadir, and plasma counter regulatory hormone and free fatty acid responses were studied in eight type I uncomplicated diabetic subjects and eight nondiabetic subjects. Each subject was tested three times at two different rates of insulin infusion (25 and 32 mU/m2/min): (1) during infusion of saline, (2) during infusion of phentolamine + propranolol (combined alpha, beta-blockade), and (3) during infusion of propranolol alone (isolated beta-blockade) for 150 min. At the time of the studies, the diabetic subjects had been made euglycemic by an overnight i.v. insulin infusion. During infusion of insulin (25 mU/m2/min) and saline, the rates of PG decrease and recovery were slower (P less than 0.01) and PG nadir was delayed in the diabetic subjects. Moreover, their plasma glucagon response was blunted while plasma epinephrine, norepinephrine, growth hormone, and cortisol responses were similar in both groups. Infusion of insulin at 32 mU/m2/min caused larger decreases in PG than had been observed when insulin was infused at 25 mU/m2/min. Plasma glucagon responses increased in the nondiabetic subjects (P less than 0.05) but not in the diabetic subjects. However, in the diabetic subjects, plasma epinephrine increased more than in the nondiabetic subjects (P less than 0.05). There was an inverse correlation between the individual plasma epinephrine responses and the plasma glucagon responses in the diabetic subjects (r = -0.72) but not in the nondiabetic subjects. Alpha, beta-adrenergic blockade decreased the plasma glucose nadir and impaired the rate at which normoglycemia was restored in the diabetic subjects (P less than 0.005 vs. saline) but not in the nondiabetic subjects. Plasma catecholamine and growth hormone responses were increased and plasma FFA recovery was suppressed in both groups (P less than 0.05 vs. saline), while the cortisol responses were unaltered. During isolated beta adrenergic blockade, changes in plasma glucose, counterregulatory hormones and FFA were essentially identical to those observed during combined alpha, beta adrenergic blockade in both groups except that the augmented plasma norepinephrine responses were no longer apparent. CONCLUSIONS: although epinephrine is not essential for prompt restoration of normoglycemia in normal man following insulin-induced hypoglycemia, it plays a major role in glucose counterregulation in diabetics who have an impaired glucagon secretion in response to hypoglycemia. These counterregulatory effects of epinephrine are mediated by beta-adrenoreceptors. PMID- 6298040 TI - Nerve function and metabolic control in teenage diabetics. AB - Peripheral somatic and autonomic nerve function have been studied in 79 teenage (16-19 yr) diabetics and 20 age- and sex-matched normal controls. Almost three quarters of the diabetics (72%) had abnormal peripheral somatic nerve function tests, and one-third (31%) had abnormal cardiac parasympathetic tests. Both motor and sensory peripheral somatic nerve abnormalities were related to poor prevailing glycemic control (HbA1) and duration of diabetes. Thus, the 27 patients with three or more (maximum six) peripheral nerve abnormalities had significantly higher HbA1 levels (P less than 0.001) and longer duration of diabetes (P less than 0.01) than the 22 with no abnormalities. Individual peripheral somatic nerve tests almost invariably correlated only with HbA1 (median motor, P less than 0.05; peroneal motor, P less than 0.001; sural sensory, P less than 0.001) or duration of diabetes (median sensory, P less than 0.001). Sensory potential amplitude, as well as conduction velocity, was frequently reduced, implying axonal involvement. These findings suggest that abnormal peripheral and autonomic nerve function are common in young insulin dependent diabetics and that poor metabolic control is a major determinant of the damage. DIABETES 32:142-147, February 1983. PMID- 6298041 TI - Failure of infused beta-hydroxybutyrate to decrease proteolysis in man. AB - Ketone bodies have been suggested to have a protein-sparing effect, since infusion of Na-beta-hydroxybutyrate in man decreases plasma alanine concentrations and urinary nitrogen (N) excretion. To test this hypothesis, six normal postabsorptive volunteers were infused with Na-beta-hydroxybutyrate for 3 h. Rates of glucose, leucine carbon, and alanine appearance and disappearance from the plasma space were traced with [3-3H]glucose, L-[6,6,6-2H3]leucine, and [2,3,3,3-2H4]alanine. Rates of leucine N appearance and disappearance and the rate of transfer of leucine N to alanine were assessed with [15N]leucine. During ketone body infusion, plasma alanine decreased (P less than 0.05), whereas plasma leucine increased (P less than 0.05). Rates of alanine appearance increased (5.3 +/- 0.3 to 7.8 +/- 0.6 mumol/kg X min), but the increase in its rate of disappearance was slightly greater, accounting for the decrease in plasma alanine concentration. Leucine N flux and the rate and percent of leucine N transferred to alanine increased, whereas leucine carbon flux was unchanged. To determine the effect of the alkalemia induced by Na-beta-hydroxybutyrate, four additional subjects were infused with NaHCO3. Alkalemia had no effect on leucine N or carbon flux or on the rate of appearance of alanine, but increased the rate of alanine disappearance, resulting in a decrease in the plasma alanine concentration. Since the rate of appearance of leucine carbon was unaltered during the infusion of Na beta-hydroxybutyrate, it is unlikely that hyperketonemia per se decreases proteolysis in postabsorptive man. PMID- 6298042 TI - Immunologic aspects of the nonobese diabetic (NOD) mouse. Abnormalities of cellular immunity. AB - Experiments were performed on 12-wk-old nonobese diabetic (NOD) mice to investigate the immunologic background of the condition, using ICR mice as controls. The results indicate the following: (1) absolute decreases in number of T lymphocytes, (2) depression of natural killer activity, (3) normal responsiveness in delayed type hypersensitivity and functional depression of killer T cells against allogeneic tumors, (4) diminished resistance to herpes virus infection, and (5) enhanced production of polyclonal antibodies to T cell dependent antigens. These features are similar to those noted in other autoimmune diseases of man and in their experimental models in laboratory animals. Elucidation of the pathogenetic mechanism of autoimmune diabetes mellitus in NOD mice, therefore, may contribute to the diagnosis, treatment, and prevention of a wide variety of autoimmune diseases. PMID- 6298043 TI - Virus-induced diabetes mellitus in mice and the thymus-dependent immune system. AB - The present study concerns the effect of the experimental diabetogenic encephalomyocarditis (EMC) virus on normal and athymic nude mice of BALB/c origin. The effect of simultaneous immunosuppressive pharmacological treatment with a derivative of cyclophosphamide in a relatively low dose (3 mg/mouse) was also studied. After inoculation with EMC virus, 36% of the normal mice, but none of the nude mice, developed diabetes mellitus and 93% of the normal mice, but none of the nude mice, developed paresis of one or more leg(s). When lower doses of EMC virus were given, few or none of the normal mice developed diabetes or paresis. After treatment with a cyclophosphamide-derivative, the number of paralysed mice increased. EMC virus in abundant amounts could be isolated from the pancreas and heart of all virus-inoculated mice, including the non-diabetic nude mice. Antibodies against EMC virus were found in all groups of virus inoculated mice, although only in small amounts in nude and immunosuppressed normal mice. Histological examination revealed no significant differences between the islets of Langerhans of the experimental mice, diabetic as well as non diabetic, and the control mice with respect to lymphocytic infiltration. It is concluded that the thymus-dependent immune system seems to be of decisive importance for the development of diabetes in this virus model. PMID- 6298044 TI - Metabolism of benzo[a]pyrene and other carcinogens in mouse epidermal keratinocytes in culture. AB - The metabolism of benzo[a]pyrene (BP) and other carcinogens was investigated in primary cultures of mouse epidermal keratinocytes isolated from the C3H/He inbred strain, a responsive strain in the Ah locus. The basal aryl hydrocarbon hydroxylase activity of these cells was 9.2 pmol equivalents of 3-hydroxy-BP per mg protein per hr. This enzyme was induced dose-dependently by benz[a]anthracene (BA) and a 7-fold increase was obtained by treatment with 13 microM BA for 24 hr. The metabolism of BP increased with the time of incubation. On thin layer chromatography, 60% of the added BP was separated as metabolites at 24 hr and 96% at 48 hr. High pressure liquid chromatography indicated that mouse epidermal cells produced almost the whole series of metabolites of BP; dihydrodiols at the 9,10-,4,5- and 7,8-positions, quinones, and phenols at the 3- and 9-positions. The metabolic activity of mouse epidermal cells was further demonstrated by a cell-mediated assay, in which V79 Chinese hamster cells were cultured on top of epidermal cells and treated with BP and other carcinogens. Mutation of V79 cells, detected as ouabain resistance, was induced in a dose-related fashion by BP, BP 7,8-dihydrodiol, methylcholanthrene, and 7,12-dimethylbenz[a]anthracene, but not by aflatoxin B1; of these compounds BP-7,8-dihydrodiol was the most potent. PMID- 6298045 TI - Inhibition of microsome-mediated benzo[a]pyrene metabolism by hemin. AB - The effects of hemin on the metabolism of benzo[a]pyrene (BP) in rat liver microsomes were studied. Hemin inhibited the activity of aryl hydrocarbon hydroxylase and the metabolic activation of BP to forms able to bind to DNA. Analyses by high performance liquid chromatography of metabolites of BP revealed that hemin reduced their production and changed their relative amounts. The decrease in the amount of BP-7,8-dihydrodiol was greater than that in the amount of phenols. The elution profile of BP-metabolite-bound deoxyribonucleosides on Sephadex LH-20 column chromatography indicated that the formation of adducts with BP-7,8-dihydrodiol-9,10-oxide was decreased by hemin more than the formation of those with phenol derivatives. The present results suggest that endogenous components that are not enzymes are important modifiers of the biological activities of chemical carcinogens. PMID- 6298046 TI - [Paraneoplastic hypercalcemia disclosing cholangiocarcinoma. Remission after hepatectomy]. PMID- 6298047 TI - A comparison of transcatheter arterial embolization with one shot therapy for the patients with hepatic cell carcinoma. AB - It has been confirmed gradually that transcatheter arterial embolization is the most effective, conservative therapy for the treatment of unresectable hepatic cell carcinoma (hepatoma). Embolization or one shot therapy was carried out in a clinical trial involving 41 patients with unresectable hepatoma visiting our department. Embolization group (emboli G): 19 cases. 1 to 6 embolizations in each case. One shot group (one shot G): 22 cases. Medications: Mitomycin C 10-40 mg and others. Disappearance rate of icterus after treatment was 50% (emboli G) and 25% (one shot G). Decrease in size of hepatomegaly or tumor was seen in 84% (emboli G) and 32% (one shot G) which was statistically significant (less than 1%). Serum AFP titer after embolization decreased in all cases but in only 5 of 12 cases (ca 41%) after one shot (less than 1%). Effective cases measured by Karnofsky's method were 18 out of 19 cases (95%) in emboli G, but in one shot G only 10 out of 22 cases (ca 45%)(less than 0.1%). Survival rate after each therapy was 67% (emboli G) and 38% (one shot G) after 6 months, and 59% (emboli G) and 19% (one shot G) at 1 year respectively. One study showed that transcatheter arterial embolization therapy was much more effective than one shot therapy. PMID- 6298049 TI - Detection of serum albumin receptor in hepatitis B virus carriers by enzyme linked immunosorbent assay. AB - The relationship between glutaraldehyde-treated polymeryzed human serum albumin (pHSA) and HBe antigen (HBeAg)-positive serum was examined by the use of a new enzyme-linked immunosorbent assay (ELISA). The author succeeded in measuring the pHSA binding activity (pHSA-BA) of HB surface antigen (HBsAg) particles in the present ELISA method using horseradish peroxidase-labelled pHSA after fixation of HBsAg on an anti-HBs-coated well of polystyrene microplates. In HBeAg-positive group, the pHSA-BA of sera of 40 asymptomatic carriers and 2 chronic persistent hepatitis (CPH) patients were higher than those of 8 chronic active hepatitis (CAH) (p less than 0.01) and 8 liver cirrhosis sera (p less than 0.05). On the contrary, in the anti-HBe-positive group the pHSA-BA of 17 asymptomatic carriers and 3 CPH sera were lower than those of 8 CAH (p less than 0.005) and 10 liver cirrhosis patient sera (p less than 0.005). In the both-negative group the pHSA BA of 8 asymptomatic carrier and 3 CPH sera were also lower than that of 8 CAH (p less than 0.05). In acute exacerbation of HBsAg-positive CAH the pHSA-BA elevated one to two months before the peak of S-GPT level, being correlated with the DNA polymerase activity. Because of its apparent reproducibility, it is concluded that low cost and some advantages may have clinical utility in the same setting as the HBeAg is now used. PMID- 6298048 TI - Enteric coated polymyxin B in the treatment of hyperammonemia and endotoxemia in liver cirrhosis. AB - Effects of enteric coated polymyxin B capsules on hyperammonemia and endotoxemia in liver cirrhosis were investigated. Six million units of polymyxin B were orally administered daily to 21 patients with liver cirrhosis and 3 patients with hepatoma cum liver cirrhosis, whose plasma ammonia was higher than normal limit and/or whose plasma endotoxin was positive, for 5-32 days, and serum polymyxin B concentration (in 5 cases), changes of plasma ammonia level (in 19 cases) and plasma endotoxin (in all cases) were observed. Serum polymyxin B concentration was below the detectable limit (0.5 unit/ml) in all cases observed. In the patients with liver cirrhosis, plasma endotoxin and ammonia levels decreased rapidly after polymyxin B treatment, and the decreases in endotoxin levels were kept throughout the treatment. Twelve patients with liver cirrhosis (10 among them were treated with lactulose) were served as controls. All patients who were treated with lactulose alone showed rapid decrease in plasma ammonia, but the decrease in endotoxin in these patients was slower than that in those treated with polymyxin B. From these results, oral administration of polymyxin B is concluded to be useful in the treatment of hyperammonemia and endotoxemia in liver cirrhosis, as a poorly absorbed antibiotic and as an antiendotoxin agent. PMID- 6298051 TI - Hepatitis B virus and primary hepatocellular carcinoma. PMID- 6298050 TI - Hepatitis B virus status of black women with hepatocellular carcinoma. AB - The reason why hepatocellular carcinoma occurs less frequently in women than in men is unknown. The possibility that differences in the hepatitis B virus status between men and women with this tumor might be important in this regard was investigated in 75 black women and 75 age- and ethnically-matched men with hepatocellular carcinoma. Prevalences of hepatitis B surface antigen, core antibody alone, and total hepatitis B virus markers were not significantly different in the two sexes. It is concluded that women with hepatocellular carcinoma are just as likely as men to be, or to have been, infected with hepatitis B virus. Because the hepatitis B virus and contraceptive steroids might conceivably act as cocarcinogens, the age distribution of women with hepatocellular carcinoma was determined. Thirty-nine percent were of child bearing age. Of these, 44% were positive for surface antigen and 17% for core antibody alone. Ninety percent of these women had one or more hepatitis B virus markers. PMID- 6298052 TI - A note on dysenzymia in the liver of rats fed on zinc, an essential metal. AB - Although zinc in traces is essential for the growth and well being of the animal, however a long-term treatment has been found equally toxic to the liver and kidney. Present report describes its effects on few key enzymes viz. alkaline phosphatase, acid phosphatase, 5-nucleotidase, lipase, glucose-6-phosphatase, and cholinesterase in the liver of rat, Rattus rattus albino. Histochemical observations have provided visual evidences on Zn-induced dysenzymia. The results have further been interpreted in terms of its effects on cellular organelle, levels of enzyme protein and microenvironment of the hepatic parenchyma. PMID- 6298054 TI - Effect of Ruscus aculeatus on isolated canine cutaneous veins. PMID- 6298053 TI - [Morphology and clinical aspects of malignant mixed mullerian tumors of the corpus uteri]. AB - Mixed muellerian tumours of the uterus are more common today than uterine leiomyosarcomata. There is a world wide increase in the incidence in the last few years. In the past 15 years we observed 20 mixed muellerian tumours of the uterine body. The patients are usually 10 years older than women with endometrial carcinoma or leiomyosarcoma of the uterus. Morphologically homologous and heterologous tumours must be differentiated. Characteristic is the marked polypoid growth. There is a close correlation between tumour stage and microscopic tumour grading. Tumours with high malignancy are usually in stage I or II. The prognosis correlates well with the grade and stage of the tumour, therefore the elaboration of a prognostic index is necessary. The survival rates are low. Mixed muellerian tumours are not at all or very little influenced by radiotherapy. The single or combined chemotherapies in use o date have not shown satisfactory success. As complete as possible surgical removal is therefore important. It cannot as yet be predicted whether our present combined treatment in stage III and stage IV by operation and post-operative combined chemotherapy with CYVADIC is liable to increase the survival rates. PMID- 6298055 TI - Reduced platelet aggregation by effects of pentoxifylline on vascular prostacyclin isomerase and platelet cyclic AMP. AB - 1. Inhibition of platelet aggregation in vitro by pentoxifylline is rather weak, requiring about 1 mM pentoxifylline. Ex vivo, however, 15 mg/kg p.o. pentoxifylline exerts an enhanced release of prostacyclin-(PGI2)-like antiaggregatory activity from rat aortas. 2. Rat aorta incubated in vitro with 1 microM pentoxifylline releases antiaggregatory activity in a similar manner. The conversion of prostaglandin H2 to PGI2-like activity which is catalyzed by vascular microsomes, also is drastically stimulated in vitro by addition of 1 microM pentoxifylline. 3. Despite its inhibitory effect on platelet cyclic AMP phosphodiesterase pentoxifylline in vitro without PGI2 has no essential effect on cyclic AMP levels in human platelets. However, in presence of PGI2, release of which probably is increased by pentoxifylline cyclic AMP level as well as inhibition of aggregation are enhanced by pentoxifylline above the effects of PGI2 itself. PMID- 6298056 TI - Sympatho-adrenal regulation of adipose tissue blood flow in dog and man. AB - 1. A comparison is made between the sympatho-adrenal regulation of subcutaneous adipose tissue blood flow in the dog and in man. 2. In the dog neuronally released noradrenaline causes vasoconstriction in subcutaneous adipose tissue by preferential activation of alpha-adrenoceptors located close to the sympathetic nerve terminals. 3. In man sympathetic reflex activation also causes vasoconstriction in subcutaneous adipose tissue, presumably via activation of alpha-adrenoceptors. 4. The adipose tissue vascular beta-adrenoceptors are of the beta 1-subtype in the dog and the beta 2-subtype in man. 5. Because of this adrenaline causes vasoconstriction in dog adipose tissue and vasodilation in human adipose tissue. PMID- 6298057 TI - Muscarinic and beta-adrenergic prejunctional modulation of adrenergic neurotransmission in the blood vessel wall. AB - Many prejunctional inhibitory and excitatory receptors have been described on adrenergic nerve endings in animal arteries and veins, and studies to date have identified some of these in human blood vessels. The latter include muscarinic receptors in cutaneous veins which when activated by acetylcholine inhibit the evoked release of norepinephrine, and beta-adrenoceptors which when stimulated by isoproterenol or epinephrine facilitate it. Animal studies suggest that cholinergic vasodilatation can result from prejunctional inhibition of adrenergic neurotransmission. The human saphenous vein appears to contain more prejunctional beta-adrenoceptors than the canine, with a consequential greater enhancement of norepinephrine release when these receptors are activated. PMID- 6298058 TI - Influence of catecholamines on gonadotropin secretion in goldfish, Carassius auratus. AB - Serum gonadotropin (GtH) concentrations in female goldfish were measured before and at various times after intraperitoneal injection of drugs altering catecholamine synthesis and neural activities. Reserpine, a depleter of neurotransmitter stores, elevated serum GtH levels compared to controls, suggesting the general involvement of neurotransmitters in altering GtH release. 6-Hydroxydopamine, a catecholaminergic neurotoxin, increased serum GtH concentration, suggesting that catecholaminergic neurons inhibit GtH release. Blocking of L-DOPA and dopamine synthesis by alpha-methyl-para-tyrosine and carbidopa, respectively, but not norepinephrine by diethyldithiocarbamate, raised serum GtH values above those of controls. Injections of an alpha-agonist, clonidine, also increased serum GtH concentrations. These results suggest the existence of an inhibitory dopaminergic and a stimulatory alpha-adrenergic influence on GtH release in goldfish. PMID- 6298059 TI - In vivo biosynthesis of melanotropins and related peptides in the pars intermedia of Xenopus laevis. AB - To study in vivo biosynthesis of pars intermedia peptides in Xenopus laevis, [3H]lysine was administered by an osmotic minipump via a cannula inserted near the pituitary gland. Following extraction of the neurointermediate lobe, high performance liquid chromatography was used to separate the newly synthesized peptides. In black-background adapted animals, [3H]lysine was incorporated into a number of peptides. The elution characteristics of these peptides corresponded exactly with those of peptides synthesized during in vitro incubation of neurointermediate lobes, and which were identified as des-N alpha-acetyl-alpha MSH, a gamma-MSH-like peptide, two corticotropin-like intermediate lobe peptides, and two forms of endorphin. In white-background adapted Xenopus, practically no synthesis of pars intermedia peptides occurred. Transfer of black-adapted toads to a white background at the beginning of infusion led to storage of newly synthesized peptides. When such animals were maintained on a white background for 10 days, des-N alpha-acetyl-alpha-MSH, but not alpha-MSH, was present in the pars intermedia; this supports the notion that des-N alpha-acetyl-alpha-MSH constitutes the "storage form" of alpha-MSH. PMID- 6298060 TI - Preparation of enriched populations of corticotrophs from goldfish rostral pars distalis. AB - A mammalian isolated adrenal cell system was validated as a bioassay for goldfish ACTH; the log dose-response curve for the goldfish hormone is parallel to that for synthetic mammalian ACTH1-24, and the two ACTHs induce the same maximum rate of corticosterone production. Using this assay, it was observed that (1) there is a marked and consistent biphasic change in pituitary ACTH content as related to the length of time the fish are held in laboratory aquaria, and (2) the absolute and relative (to tissue wt) ACTH content of the rostral pars distalis is considerably greater than that of the proximal portion of the gland. Using a trypsin technique, isolated pituitary cells were prepared from both (rostral and proximal) portions of the gland; bioassay data indicate that cell suspensions prepared from the rostral pars distalis are enriched with respect to corticotrophs. The implications of these findings with regard to formulating an advantageous in vitro system for studying ACTH secretion are discussed. PMID- 6298061 TI - Urinary cyclic AMP excretion in birds: dependence on parathyroid hormone activity. AB - The cyclic AMP response of avian kidney to parathyroid activity has been evaluated both in vitro and in vivo. The production of cyclic AMP by dispersed avian kidney cells was stimulated by bovine parathyroid hormone or by an extract of avian parathyroid glands. Intravenous infusion of bovine parathyroid hormone resulted in increased urinary excretion and plasma concentration of cyclic AMP, as well as increased plasma calcium and urinary phosphorus excretion. The increases in plasma and urinary cyclic AMP preceded those of plasma calcium and of phosphorus excretion. EDTA infusion resulted in a decrease in plasma calcium and an increase in urinary cyclic AMP excretion. After 10 days on a low-calcium diet, chickens exhibited a 4.5-fold increase in urinary cyclic AMP excretion. The results suggest that urinary cyclic AMP in chickens reflects PTH activity similar to some mammalian species. PMID- 6298062 TI - The plasmid as a tool for plasmid genetics. II. Isolation of point mutations that affect replication of a ColE1-related plasmid. PMID- 6298063 TI - Nostoc PCC7524, a cyanobacterium which contains five sequence-specific deoxyribonucleases. AB - Five nucleotide sequence-specific deoxyribonucleases present in cell-free extracts of the filamentous cyanobacterium Nostoc PCC7524 have been purified and characterized. One of these enzymes, designated Nsp(7524)I cleaves at a new kind of nucleotide sequence, i.e. 5'-PuCATG Py-3'. The other four restriction enzymes in this organism, designated Nsp(7524)II, Nsp(7524)III, Nsp(7524)IV and Nsp(7524)V, are isoschizomers of enzymes which have been previously described. The cleavage site of Nsp(7524)II which is an isoschizomer of SduI was determined. PMID- 6298064 TI - A new approach for molecular cloning in cyanobacteria: cloning of an Anacystis nidulans met gene using a Tn901-induced mutant. AB - A new strategy for molecular cloning in the cyanobacterium Anacystis nidulans R-2 is described. This strategy involved the use of a transposon and was developed for the cloning of a gene encoding methionine biosynthesis. A met::Tn901 mutant was isolated. Chromosomal DNA fragments were cloned in the Escherichia coli plasmid vector pACYC184. A recombinant plasmid carrying the inactivated met::Tn901 gene was selected after transformation to E. coli. The cloned met::Tn901 DNA fragment was used as a probe to select the corresponding A. nidulans R-2 wild-type met gene from a gene library prepared in E. coli, using the newly constructed shuttle cosmid vector pPUC29. When transformed into A. nidulans Met- mutants, this cloned gene allowed the mutants to grow prototrophically. PMID- 6298065 TI - Ethidium-bromide-inhibited restriction endonucleases cleave one strand of circular DNA. AB - We analyzed the effect of ethidium bromide (EtBr) on the cleavage of closed circular pBR322 DNA molecules by six restriction enzymes which make staggered or flush cuts (EcoRI, HindIII, BglI, PstI, HincII, PvuII). EtBr concentrations and reaction temperatures were determined at which DNA molecules with single-strand breaks were the major reaction product of digestion by all the enzymes. However, the amounts of intermediates which could be isolated differed for various enzymes. The results extend previous studies, showing that sequential cleavage of the DNA strands probably is a general property of restriction endonucleases. PMID- 6298066 TI - Physical analysis of antibiotic-resistance genes from Streptomyces and their use in vector construction. AB - Restriction endonuclease cleavage maps of five DNA fragments carrying genes for neomycin phosphotransferase and neomycin acetyltransferase (from Streptomyces fradiae), viomycin phosphotransferase (from S. vinaceus), and ribosomal methylases determining resistance to thiostrepton (from S. azureus) and MLS antibiotics (from S. erythreus) are described, together with a map for the SLP1.2 Streptomyces plasmid used to isolate the fragments. Construction of a versatile Streptomyces cloning vector (pIJ61) is reported. pIJ61 carries neomycin phosphotransferase and thiostrepton resistance genes and has unique BamHI and PstI sites which will allow clone recognition by insertional inactivation of neomycin resistance; cloning sites for several other endonucleases are also present. pIJ28, a shuttle vector for Streptomyces and E. coli, carries neomycin resistance and the SLP1.2 and pBR322 replicons. PMID- 6298068 TI - An IS4 transposition causes a 13-bp duplication of phage lambda DNA and results in the constitutive expression of the cI and cro gene products. AB - We have examined the nature of the additional DNA present in lambda hyp- mutants (Eisen et al., 1982). This DNA is an IS4 element in orientation I, in the y region of bacteriophage lambda at nucleotide position 39,139 (see Moore et al., 1979). Our assignment is based on (i) the similarity in size derived from the PstI, AvaI, and HindII restriction pattern and (ii) the DNA sequence of both the left and right lambda-IS4 DNA junctions in phage lambda hyp15rev4. The IS4 integration event resulted in the duplication of 13 bp of lambda DNA in contrast to the 11- and 12-bp duplications previously observed at the sites of IS4 integrations elsewhere (Klaer et al., 1981). PMID- 6298069 TI - Management of common non-malignant GI problems in the elderly. PMID- 6298067 TI - Mutants in the y region of bacteriophage lambda constitutive for repressor synthesis: their isolation and the characterization of the Hyp phenotype. AB - Bacteriophage lambdahyp mutants have been isolated as survivors of Escherichia coli K-12 bacteria lysogenic for lambda Nam7am53cI857. The hyp mutants are characterized by (i) their localization in the y region very close to the imm lambda/imm434 boundary, (ii) polarity on O gene expression, (iii) immediate recovery of lambda immunity at 30 degrees C after prolonged growth of lambda Nam7am53cI857 hyp lysogens at 42 degrees C even in the presence of an active cro gene product, (iv) ability of phage lambda v2v3vs326 but not lambda v1v2v3 to propagate on lambda cI+hyp lysogens, (v) inability to express lambda exonuclease activity after prophage induction, and (vi) inviability at any temperature of phage carrying the hyp mutation. All these properties are referred to collectively as the Hyp phenotype. We show that the Hyp phenotype is due to cII independent constitutive cI-gene-product synthesis originating in the y region, which results in the synthesis of anti-cro RNA species, and constitutive levels of cro gene product present even in lambda cI+hyp lysogens. A model is presented which is consistent with all the experimental observations. PMID- 6298070 TI - [Improved method of indicating enteroviruses in the soil by using ultrasound]. PMID- 6298071 TI - [Cyclic AMP in trophoblast homogenates. III. The trophoblast]. PMID- 6298072 TI - [3 epithelial cancers treated successively in 1 patient over a 10-year period]. PMID- 6298073 TI - Drugs and the elderly. Antiulcer agents. PMID- 6298074 TI - The role of phospholipids in the binding of oxytocin to its receptors in lactating rabbit mammary gland. AB - Phospholipase C (Clostridium welchii and Bacillus cereus) treatment of lactating rabbit mammary gland membranes (140,000 g pellet and sucrose density gradient purified plasma membranes) resulted in a large decrease in the binding of [3H]oxytocin to these subcellular fractions. This decrease was not due to a solubilization of oxytocin receptors but was the result of the removal of phospholipids which may participate in the hormone-receptor interaction. Phospholipase C treatment of the membrane fractions resulted in a dose-dependent removal of different classes of phospholipids. Sphingomyelin, phosphatidylcholine and phosphatidylethanolamine were removed by phospholipase C (C. welchii and B. cereus) treatment. No significant change was observed in the content of phosphatidylinositol. Phospholipase C from B. cereus reduced the content of phosphatidylserine, while the enzyme from C. welchii did not. PMID- 6298075 TI - [Glomus tumor of the digits]. PMID- 6298076 TI - [Metastatic small cell pulmonary carcinoma presenting as prostatic obstruction]. PMID- 6298077 TI - [The laboratory diagnosis of infections associated with Epstein-Barr Virus]. PMID- 6298078 TI - [Combined chemotherapy and radiotherapy for localized small cell pulmonary carcinoma]. PMID- 6298079 TI - [Platelets and their products in coronary spasm]. PMID- 6298080 TI - [Anaphylactoid reactions to iodine containing contrast agents]. PMID- 6298081 TI - Cytochemical localization of some hydrolases in the pollen and pollen tubes of Amaryllis vittata Ait. AB - Some hydrolases are localized cytochemically in the pollen and pollen tubes of Amaryllis vittata Ait. The function of different enzymes is discussed in relation to pollen tubes morphogenesis. Activity of most of the enzymes was confined to colpus region, pollen wall and general cytoplasm of pollen and pollen tube. The activity of hydrolytic enzymes like acid monophosphoesterase and lipase and was nil in the exine of both germinated and ungerminated pollen, whereas intense reaction for esterase was observed in exine. Enzyme activity increased after germination which suggest the hydrolysis of stored metabolites and synthesis of proteins and other metabolites for the active growth of pollen tube. Intense reaction for enzymes like alkaline phosphomonoesterase, ATP-ase, 5-nucleotidase etc. at the tip region of pollen tube suggest their role in physiological processes associated with exchange of materials through intercellular transport during tube wall polysaccharide biogenesis. PMID- 6298082 TI - [Potentiative effects of angiotensin converting enzyme inhibitors on carrageenan induced edema in rats]. AB - Carrageenan-induced edema of rat paw was greatly potentiated by orally or locally administered angiotensin converting enzyme (CE, identical with kininase II) inhibitor, (4R)-3-[(2S)-3-mercapto-2-methylpropanoyl]-4-thiazolidinecarboxylic acid (YS980). The potentiative effect of YS980 was observed by the administration not only prior to but also 3 hr after the carrageenan injection. The vascular permeability test showed the same potentiative effect of YS980 in the inflamed tissue of carrageenan edema. Bradykinin potentiating peptide-B (BPP-B) and 1,10 phenanthroline also potentiated the edema and the effects of these compounds were in order of YS980 greater than BPP-B greater than 1,10-phenanthroline. The order was in accordance with that of inhibitory potencies against kininase II activity of rat serum in vitro. However, the activity of kininase I was not affected by YS980 and BPP-B. In addition, among various experimental models of acute paw edema, the potentiative effects of YS980 were restricted to the inflammations mediated by kinins such as carrageenan, cellulose sulfate, kaolin and bradykinin induced edema. These results suggest that the potentiative effect of YS980 on carrageenan edema is mainly based on its inhibitory effect on kininase II in the inflamed site. PMID- 6298083 TI - Metabolic profile of caecal micro-organisms from rats fed indigestible plant cell wall components. AB - A fibre-free diet, or the same diet supplemented with 100 g cabbage or carrot cell-wall preparation/kg, was fed to rats for 28 days and the activities of a number of caecal microbial enzymes (azoreductase, aryl nitroreductase, beta glucosidase, beta-glucuronidase, imidazole nitroreductase and nitrite reductase) were determined in vitro. The plant cell-wall preparations diluted the gut contents and decreased the number of bacteria per gram of caecal contents. Enzyme activities per gram of caecal contents were also decreased, with the exception of beta-glucosidase activity which was significantly increased. These plant cell wall preparations also increased caecal size, and thereby significantly increased total activity per caecum of microbial azoreductase, aryl nitroreductase, beta glucosidase and beta-glucuronidase. When bacterial metabolism was expressed per 10(9) bacteria, all enzyme activities were significantly increased in caecal samples from rats fed the plant cell-wall preparations. There was an overall concordance of 0.91 between all the enzymes when expressed per 10(9) bacteria, but of only 0.38 when enzyme activities were expressed per gram of caecal contents. PMID- 6298085 TI - [Circumscribed intra-areolar skin edema in cranially localized breast cancer]. PMID- 6298086 TI - [Oral treatment of chronic recurring vaginal candidiasis with Nizoral]. PMID- 6298084 TI - Protection by indole-3-carbinol against covalent binding of benzo[a]pyrene metabolites to mouse liver DNA and protein. AB - Indole-3-carbinol (I-3-C) is a compound present in many cruciferous vegetables that has been shown to reduce aryl hydrocarbon-induced neoplasia in experimental animals. We examined the relationship between the ability of I-3-C to alter the activity of hepatic aryl hydrocarbon hydroxylase (AHH), and its ability to inhibit the covalent binding of benzo[a]pyrene (BaP) metabolites to DNA and protein. Using an in vitro system and a hepatic postmitochondrial fraction from mice that had been treated by gavage with I-3-C, we found that up to 90% of the covalent binding of BaP metabolites to macromolecules was eliminated, while AHH activity was unchanged. In experiments in vivo, treatment of mice by gavage with I-3-C before [14C]BaP resulted in up to an 80% decrease in covalent binding of 14C to DNA or protein with no concomitant decrease in hepatic AHH activity. These results suggest that I-3-C administered in vivo confers protection against the binding of BaP oxidation products to hepatic cellular macromolecules. PMID- 6298087 TI - [Analysis of macrophage function of tumor bearing mice]. PMID- 6298088 TI - The depot-form of glucocorticoid decreases cortisol but not ACTH and beta endorphin levels in human plasma. PMID- 6298089 TI - Verapamil and induced cortisol, aldosterone, TSH secretion. PMID- 6298090 TI - Ontogenetical variations of transcortin modulate glucocorticoid receptor function and corticotropic activity in the pituitary gland. AB - This study was aimed at investigating the influence of tissue and plasma transcortin on pituitary glucocorticoid receptor function and corticotropic activity during development. "In vivo" nuclear uptake of (3H)corticosterone was found to be increased in pituitary glands from newborn rats which, unlike that of adults, lack transcortin-like material. Immature glands incubated "in vitro" likewise showed enhanced nuclear accumulation of tracer; the magnitude of uptake being inversely related to both tissue and plasma contents of transcortin. Competition studies indicate that pituitary transcortin modulates interaction of corticosterone with receptor binding sites and, hence, may interfere with steroid translocation to the nucleus. Moreover, we found that steroid-induced inhibition of ACTH release was highest in those pituitaries which were deficient in transcortin-like molecules. We conclude that the binder plays a modulating role in pituitary glucocorticoid receptor function. PMID- 6298091 TI - Marathon run: effects on plasma renin activity, renin substrate, angiotensin converting enzyme, and cortisol. AB - Altogether 33 Finnish amateur runners were studied before and after a non competitive Marathon run over the classical itinerary in Athens in 1976 (n = 8), 1977 (n = 14) and 1978 (n = 11). Plasma renin activity (PRA) rose 3-fold in all runs, whereas plasma renin substrate (RS) concentration did not change significantly. Serum angiotensin converting enzyme (ACE) activity was not changed. Serum cortisol concentration was increased 2-3 fold. The unchanged plasma RS concentration, in spite of increasing PRA, indicates that plasma RS is kept within normal limits during prolonged strenuous physical exercise. One contributing mechanism may be stimulation of RS biosynthesis by cortisol. Low PRA levels in two old runners, 65 and 83 years old, may indicate a decreased ability to respond with renin release to the stimuli of physical exercise. PMID- 6298092 TI - Histochemically demonstrable enzyme activities and their independence of the hormone receptor content in female breast carcinoma. AB - Acid phosphatase, leucine aminopeptidase, monoamine oxidase and non specific esterase activities were histochemically demonstrated in specimens derived from 15 infiltrating ductal carcinomas of female breast. The relative areas occupied by the enzyme-positive carcinoma cells were visually estimated and, in the cases of leucine aminopeptidase, assessed morphometrically. All enzyme activities were found to be subject to major variations within a single carcinoma and between individual carcinomas, and the activity of any single enzyme was independent of that of three others. None of the enzyme activities correlated with the estrogen and progesterone receptor values, nor the histological grade of malignancy of the tumour. Thus, histochemically demonstrable enzyme activities seem to be of no use in predicting the hormone receptor content in infiltrating ductal carcinomas of the female breast. PMID- 6298093 TI - Deinstitutionalization and the private general hospital inpatient unit: implications for clinical care. AB - Deinstitutionalization has led to a rapid shift from reliance on state hospitals to use of community-based inpatient psychiatric services. While these inpatient units were initially envisioned as an integral part of the community mental health system, a number of sociopolitical and clinical pressures have caused general hospitals to respond to their new responsibilities in different ways. The authors review trends in deinstitutionalization, the diverging interests of public and private general hospitals, and problems in patient care that result. Based on a discussion of how Beth Israel Hospital, a nonprofit private general hospital in Boston, has dealt with such issues, they describe adaptive responses in four areas: admission criteria, patient management approaches, aftercare planning, and staff training. PMID- 6298094 TI - Polymorphism of the Hinf I restriction site located 1 Kb 5' to the human beta globin gene. AB - Mapping of the DNA from 14 Mediterranean subjects indicates a genetic variation in an Hinf I recognition site located 1 kilobase 5' to the beta-globin gene. This Hinf I site was found associated with eight beta-thalassemic genes and 11 normal beta genes, and hence is not specifically linked to beta-thalassemia. PMID- 6298095 TI - Mapping the alpha-globin genes in Hb J Mexico carriers. AB - The organization of the alpha-globin genes was studied by restriction endonuclease mapping, in subjects carrying the alpha variant Hb J Mexico. A subject homozygous for Hb J synthesized both Hb J (about 55%) and Hb A and had two alpha loci per chromosome. His restriction site map was found to be identical to that obtained with a normal DNA, except for a mutant Bgl II site which was observed on the Hb J chromosome proximal to the 5' alpha-locus. We have also mapped the DNA of a compound heterozygote for Hb J and alpha-thalassemia, who synthesizes 38% Hb J and we have found a single alpha gene corresponding to a - alpha 3.7 haplotype on one chromosome and two alpha genes, respectively alpha J and alpha A, on the other. PMID- 6298096 TI - A Hind III restriction site polymorphism in the human collagen alpha 1 (I)-like gene on chromosome No. 7. PMID- 6298097 TI - Inhibitory action of Polymyxin B on Ehrlich Ascites Carcinoma. PMID- 6298098 TI - beta-Adrenergic receptors of cultured mouse epidermal cells (HEL/37). PMID- 6298099 TI - Influence of non-MHC T lymphocyte alloantigens on regression of Rous sarcomas in the chicken. AB - Chickens of Regional Poultry Research Laboratory (RPRL) inbred line 6(3) regress sarcomas induced by Bryan high-titer Rous sarcoma virus to a greater extent than chickens of line 7(2), although these lines are identical for the major histocompatibility complex (MHC, B complex). They differ, however, at two independent autosomal loci, Ly-4 and Th-1, which determine surface alloantigens of partly overlapping subsets of T lymphocytes. Association of genotypes at these loci with quantitative variation in ability to regress Rous sarcomas was tested in segregating progeny derived from crosses of lines 6(3) and 7(2). In the F4 generation chickens of the Ly-4a/Ly-4a, Th-1a/Th-1a genotype (symbolized aa/aa) had significantly higher regressor ability than any of the other three double homozygous genotypes. In F5, all nine genotypes formed by combinations of homozygotes and heterozygotes were tested, and higher regressor ability was shown by the aa/aa, ab/aa, and aa/ab genotypes. These results indicate that higher regression is associated with: (1) interaction between the line 6(3) Ly-4a and Th 1a alleles in homozygous form; and (2) dominance x dominance interaction, in that the a allele at each locus is dominant for higher regression only within the homozygous aa genotype at the other locus. PMID- 6298100 TI - Renal vasodilation by converting enzyme inhibition. Role of renal prostaglandins. PMID- 6298102 TI - Beta adrenergic receptor response coupling in hypertrophied hearts. AB - Reports in the literature have indicated that inotropic responsiveness to isoproterenol (ISO) of hypertrophied rat myocardium is decreased. We have studied possible biochemical mechanisms to explain this. Adenylate cyclase activity in myocardial membranes from 13-week-old spontaneously hypertensive rats (SHR) was unchanged compared to enzyme activity in Wystar-Kyoto (WKY) when measured under basal conditions or following NaF, a guanosine triphosphate analog, MnCl2, or forskolin stimulation. However, ISO-stimulated cyclase activity was decreased as were beta-adrenergic receptor density with no change in receptor affinity. On the other hand, hearts from two-kidney, one clip renal hypertensive rats 6 weeks after initiation of hypertension showed decreased basal, ISO, NaF, and GTP analog stimulated cyclase activity with no change in Mn++ or forskolin-stimulated activity. In this model, receptor density increased. When the clipped kidney was removed, these changes returned toward normal as did the blood pressure and heart weight. We interpret these data to indicate that in SHR the biochemical defect leading to decreased inotropic responsiveness of hypertrophied hearts is due to decreased beta-adrenergic receptor density, leading to decreased ISO-stimulated cyclase activity. The nucleotide regulatory protein component (N) and the catalytic subunit (C) are not changed. In the renal hypertensive rat, on the other hand, although the C is unchanged, there is a decrease in the activity of N, and this is probably the primary reason why inotropic responsiveness to ISO is decreased. Increases in receptor density seen in this model may possibly be compensatory. PMID- 6298101 TI - Presynaptic alpha- and beta-adrenoceptor stimulation and norepinephrine release in the spontaneously hypertensive rat. PMID- 6298103 TI - Reflex-hemodynamic adjustments and baroreflex sensitivity during converting enzyme inhibition with MK-421 in normal humans. PMID- 6298104 TI - beta-Receptors and contractile reserve in left ventricular hypertrophy. AB - The inotropic responsiveness to adrenergic stimulation is diminished in hypertension associated with left ventricular hypertrophy (LVH). This was shown in hypertrophied hearts from renal hypertensive rats (RHR) (two-kidney, one clip hypertension, Goldblatt) 6 weeks after renal artery clipping when compared to age matched normotensive sham-operated controls (NR). The isoproterenol-stimulated inotropic responses (delta peak dP/dt) of isolated hearts perfused at constant pressure were significantly lower in RHR than in NR (p less than 0.001 by analysis of variance and covariance, including repeated measures). This reduction in ventricular responsiveness of isolated hearts from RHR did not extend to other inotropic agents such as calcium ions and the cardiotonic cardiac glycoside scillaren. Assay of beta-adrenergic receptors by binding to (-) [3H] dihydroalprenolol showed that left ventricular beta-receptor numbers (fmol per mg membrane protein) were significantly reduced in RHR compared to NR (28.2 +/- 1.1 vs 36 +/- 2.6, p less than 0.01) with no significant change in affinity (Kd,nM) (1.9 +/- 0.27 vs 2.26 +/- 0.34,NS). The results of this study suggest that LVH in renovascular hypertension is associated with impairment in inotropic responsiveness to beta-receptor stimulation parallel with and, in part, related to, a reduction in ventricular beta-receptor concentrations. Such blunting of inotropic responsiveness to beta-adrenergic stimulation may be one of the mechanisms in the progression from LVH to heart failure in hypertensive disease. PMID- 6298106 TI - Atrial natriuretic factor. Characterization and partial purification. AB - One of the main differences between atrial and ventricular cardiocytes is the presence in former of specific granules with morphological characteristics very similar to secretory granules found in peptide-secreting endocrine cells. It has been suggested that these granules are the storage place for the atrial natriuretic factor. In the rat, water deprivation produces an increase in atrial granularity but a significant decrease in acid-extractable diuretic and natriuretic activity, suggesting that the number of atrial specific granules does not necessarily represent natriuretic activity. The atrial natriuretic factor activity is destroyed by incubation with several proteases and does not inhibit the sodium-potassium ATPase, suggesting that the active substance is a small peptide that is probably different from the so-called natriuretic hormone. After a series of chromatographic steps in Sep-Pak cartridges, Bio-Gel P-10, CM Bio Gel, and Mono S columns, the specific activity of the atrial natriuretic factor was increased from 193, corresponding to atrial homogenates, to 242,000, which corresponds to the last chromatographic step representing a 1250-fold purification. This material showed a potent natriuretic activity, as 10 picomoles increased natriuresis by 100%. PMID- 6298105 TI - Calcium balance and parathyroid hormone mediated vasodilation in the spontaneously hypertensive rat. PMID- 6298107 TI - Effect of anteroventral third ventricle lesions on vascular sodium-pump activity in two-kidney Goldblatt hypertension. AB - We studied the effects of anteroventral third ventricle (AV3V) lesions on the vascular Na+-pump activity and blood pressure of rats prepared by the two-kidney Goldblatt procedure. Blood pressures and Na+-pump activity of the isolated tail arteries, measured as ouabain-sensitive 86Rb+-uptake were determined in rats with renal artery clips, rats with AV3V lesions, and rats with AV3V lesions. Rats with renal artery clips developed higher blood pressures (40%) and higher vascular Na+ pump activity (20%-35%) than rats with no renal clips. Placement of AV3V lesions prior to the placement of renal clips prevented the increase in blood pressure and the increase in vascular Na+-pump activity. Plasma potassium and creatinine concentrations, nonspecific 86Rb+-uptake, and hematocrit were not different among these groups. Plasma sodium concentration was elevated in the AV3V lesioned control group. These experiments suggest a possible role of this CNS region in the regulation of vascular Na+-pump function during hypertensive states. PMID- 6298108 TI - Sodium-potassium pump activity in reduced renal-mass hypertension. PMID- 6298109 TI - Characterization of pulmonary cellular influx differentials to known toxic agents between species. AB - In this study, we have shown that chickens, frogs, and toads are resistant to acute pulmonary injury by a variety of toxic agents, (O2, hyperbaric O2, paraquat, and silica), that cause extensive acute injury in mammals. Acute pulmonary injury is defined as a massive influx of inflammatory cells, both interstitially and into the alveolar spaces, pulmonary edema, hemorrhage, and the presence of H2O2 and O-2 in the lavaged supernatant, occurring within 48 h. In some cases, chronic effects of the toxins were observed after 90 h., i.e., hemorrhage, fibrosis, and an accumulation of interstitial inflammatory cells. In all three nonmammal systems, isolated inflammatory cells failed to respond chemotactically in vitro to known mammalian chemotaxins. Pulmonary lavage of the exposed chickens, frogs, and toads also failed to produce inflammatory cells. Pulmonary edema was not detected in any of the animals by comparison of lung weight to total body weight. Intratracheal injections of silica for 2 weeks did produce chronic effects in chickens and frogs. Morphologically, the lungs showed signs of fibrosis and accumulation of interstitial inflammatory cells, but no intraalveolar cells. After 90 h of hyperbaric O2, frogs exhibited a massive infiltration of interstitial inflammatory cells and hemorrhage. Elevated O2 levels (100%) for 2 weeks under normal atmospheric conditions produced no changes in frog lungs or in the amount of inflammatory cells in the lungs. Intravenous injections of paraquat for up to 208 h failed to initiate an accumulation of pulmonary inflammatory cells or the development of pulmonary edema in chickens. There was also no detectable H2O2 or O-2 in the lavaged supernatant. It was not determined whether paraquat had a longer or more chronic effect on chickens. We suggest that the lack of an acute pulmonary inflammatory mechanism in chickens, frogs, and toads is in part responsible for the resistance of these animals to acute pulmonary injury by oxidizing mammalian toxins. PMID- 6298111 TI - Visceral leishmaniasis in Kenyan children. PMID- 6298110 TI - Effect of deuterium oxide on neutrophil oxidative metabolism, phagocytosis, and lysosomal enzyme release. AB - We have previously shown that deuterium oxide (D2O) enhances the oxidation of methionine, a myeloperoxidase (MPO) -mediated reaction, by human neutrophils during phagocytosis. However, D2O has no effect on the oxidation of methionine by the purified MPO-H2O2-Cl- system. To explain this observation, we studied the effect of D2O on the oxidative metabolism, phagocytosis, and lysosomal enzyme release by human neutrophils. D2O stimulated the hexose monophosphate shunt (HMS) activity of resting neutrophils in a dose-response fashion. In the presence of latex particles or phorbol myristate acetate (PMA), D2O brought about an exaggerated stimulation of the HMS activity. This enhancement of the HMS activity by D2O was markedly reduced when neutrophils form two patients with X-linked chronic granulomatous disease (CGD) were used, either in the presence or absence of latex particles or PMA. Superoxide and H2O2 production by neutrophils in the presence of latex particles or PMA were also stimulated by D2O. In contrast, D2O inhibited the ingestion of latex particles. D2O enhanced the extracellular release of MPO, but not lactate dehydrogenase, by neutrophils only in the simultaneous presence of cytochalasin B and latex particles. The enhancement of HMS activity and MPO release by D2O was partially inhibited by colchicine. Our results suggest that enhancement of neutrophil oxidative metabolism by D2O may in part explain the stimulation of methionine oxidation by phagocytosing neutrophils. PMID- 6298112 TI - [Use of an electronic gravity infusion regulator for infusion therapy in routine conditions. Comparative clinical study]. AB - In comparison with the ordinary roller clamp the gravity-infusion controller allows a significant lowering of complications and start-ups at peripheral venous access sites. These results are effectively based on a decrease in the infiltration rate of about 30%. For approximately 80% of the remaining complications an early alarm and interruption of the infusion takes place. It is of importance that the electronic controller avoids the excessive variations of the drop rate found with roller clamps. In contrast to these advantages problems may arise under certain circumstances caused by the narrowing of the control range based on the limited available hydrostatic pressure. Application of the electronic gravity controller must be seen primarily for peripheral venous access sites. Its use is particularly advantageous where an infiltration of the infusion solution results in an endangering of the patient (cytostatic agents, strong alkaline solutions, for instance NaHCO3, potassium concentrates, medication additions, for instance vasoconstricting agents). In addition, it makes sense to use the controller in situations where the accuracy of an infusion regulated by a roller clamp is considered to be inadequate. PMID- 6298113 TI - Changes of membrane fluidity in erythrocytes of lead-exposed workers. AB - We have studied the effect of lead on the fluidity of erythrocyte membrane to clarify if lead can interact in vivo with biological membranes. Erythrocyte membranes were chosen in our study because a decrease of red cell osmotic fragility is also evident in the absence of laboratory and clinical signs of anaemia. The study was undertaken using the Electron Spin Resonance technique with two spin labels 5-doxyl stearate and 16-doxyl-stearate, which probe the physical state of the polar surface and the inner core of the membrane respectively. Red blood cells and erythrocyte ghosts were prepared from the blood of workers occupationally exposed to lead and from healthy controls. The determinations of Pb blood, Pb urine, urine coproporphyrin and delta aminolevulinic acid showed an increased internal dose of lead, but the ordinary metabolic and haematological parameters were in the normal range. Our results show that in lead workers there is a change in chemical physical state both in erythrocytes and erythrocyte ghosts consistent with a decrease of membrane fluidity, which is evident in the surface as well as in the inner core of the membrane. The degree of membrane fluidity modification does not appear correlated with blood lead level. Changes in the membrane structural organization could be the molecular basis of some pathological alterations induced by lead. PMID- 6298115 TI - Adenylate cyclase, cyclic AMP and IgE-mediated desensitization in rat mast cells. AB - Rat peritoneal mast cells were desensitized up to 100% with 1-4 additions of suboptimal concentrations of anti-rat IgE over 60-90 min. Desensitized cells rechallenged with anti-IgE showed a 2- to 10-fold increase in cAMP during the initial 1-5 min. Membrane preparations from desensitized cells showed up to a 100 fold rise in cyclase activity following incubation with anti-IgE, compared to a 5 to 10-fold rise in control cells. Control and desensitized cell membranes rechallenged with anti-IgE showed nearly identical levels of phosphodiesterase activation. We conclude that following desensitization, repetitive low levels of anti-IgE binding induce atypically high and sustained activation of adenylate cyclase which may reflect primary or secondary regulatory events in the development of desensitization. PMID- 6298114 TI - Overall prognosis of patients with diagnosed chronic organic solvent intoxication. AB - The prognosis of 87 patients (mean age 38.6 years) was examined who had been diagnosed 3-9 years earlier as having chronic organic solvent intoxication due to trichloroethylene, perchloroethylene, or a mixture of solvents. The methods comprised an interview, a clinical neurological examination, neurophysiological examinations (EEGs and electroneuromyographs), and a psychological examination. In order to assess the prognosis, the patient's condition at the time of diagnosis was evaluated from the hospital records, and the condition upon reexamination was compared to that at the time of diagnosis. At the time of diagnosis, 31 patients had objective clinical signs in the neurological examination, whereas the remaining 56 had only subjective symptoms and neurophysiological or psychological findings attributed to the slight solvent intoxication. Upon reexamination, 42 patients had clinical neurological signs. Based on the clinical overall evaluation, the condition of 21 patients had deteriorated during the follow-up period, the condition of 23 had improved, and that of 43 had remained unchanged. The prognoses of the results of the separate examinations correlated poorly with each other. No statistically significant correlation was found between the overall prognosis and age, sex, the duration and the level of exposure, the termination of exposure after diagnosis, the presence of other diseases, or the use of alcohol. PMID- 6298116 TI - Guanine nucleotide mediated desensitization of adenylate cyclase in cell free preparations from a Leydig cell tumour. AB - Cell free desensitization of a tumour Leydig cell plasma membrane adenylate cyclase has been demonstrated in the presence of guanine nucleotides. In experiments in which the membranes were pre-incubated with various nucleotides and LH, it was shown that this decreased adenylate cyclase activity was dependent on the presence of GTP and occurred both in the presence and absence of ATP. While pre-treatment with LH alone appeared to enhance subsequent adenylate cyclase activity, this hormone was able to potentiate the desensitizing effect of GTP. The desensitizing effect of GTP was not inhibited by sodium fluoride. In contrast, the GTP analogue p(NH)ppG (guanosine 5'beta, gamma-imido triphosphate) caused a persistent activation of the adenylate cyclase. GMP and guanosine also initially inhibited the adenylate cyclase activity, but this was entirely reversed by p(NH)ppG plus LH. GDP in addition to GTP caused desensitization but this was only partially reversed by p(NH)ppG plus LH. It is proposed that in similarity with the ovary (Bockaert et al. 1976; Ezra & Salomon 1982a) desensitization of Leydig tumour cell plasma membrane adenylate cyclase may involve a GTP-mediated phosphorylation step. PMID- 6298117 TI - The use of organic substances as emergency dosimeters. AB - In unexpected encounters with ionizing radiation, it is possible to monitor absorbed exposure or absorbed dose by ESR analysis of incidental organic materials carried by both workers and non-workers with radiations. Materials of interest include plastics, paper, clothing, buttons, human hair and nail. The concept is to correlate with dose the quantity of free radicals generated in organic materials by irradiation. The experiments were aimed at determining the sensitivity, the fading, the effects of differences in conditions of storage such as temperature, illumination, and washing before ESR analysis. The method should prove useful for estimating the extent of radiation exposure at strategic locations on the body. PMID- 6298118 TI - A monoclonal antibody marker of human primitive endoderm. AB - Monoclonal antibodies were raised against the human germ-cell tumor line, LICR LON HT39/7. The cells of this human embryonal carcinoma cell line were first fixed in formol saline and then dehydrated prior to immunization so that antibodies were produced which reacted with antigens surviving conventional histological processing. Eleven hybrid clones secreted antibody that reacted with both formol saline-fixed and living cells of the immunogen. One of these antibodies, LICR LON FC 10.2, has been found to react with a membrane glycoprotein of molecular weight 200,000 restricted in distribution to the surfaces of undifferentiated teratoma-derived cell lines, the luminal surface of human fetal intestine and bronchus and areas of putative endodermal differentiation within germ-cell neoplasms. PMID- 6298119 TI - Cell-mediated immunity to Epstein-Barr virus (EBV) and natural killer (NK)-cell activity in the X-linked lymphoproliferative syndrome. AB - The activity of T-cell-mediated immunity to Epstein-Barr virus (EBV) was assessed by an assay of regression of the outgrowth of EBV-infected autologous B cells. Regression and natural killer (NK)-cell activities were compared for patients and their mothers from five families with X-linked lymphoproliferative syndrome (XLP) and three control groups. Seven of the 10 patients with XLP exhibited weak T-cell activity against autologous EBV-infected lymphoblastoid cell lines (LCL) comparable to EBV-seronegative controls. In contrast, 8 of 10 obligate carrier females of XLP had unusually strong activity, which was comparable to anti-early antigen (EA) positive controls. The results of the regression assays correlated with their EBV serology: mothers showed high titers, and their affected sons showed low-titer EBV-specific antibody responses. Defective NK-cell activity was found only in the patients with XLP. NK and regression activities did not correlate. Our findings explain, in part, the vulnerability to EBV of males with XLP and why their mothers are protected from life-threatening phenotypes of XLP. PMID- 6298120 TI - Effect of epidermal growth factor on clonogenic growth of primary human tumor cells. AB - The effect of EGF on the soft agar colony-forming ability of fresh human tumor cells was assessed in 40 specimens obtained from various types of carcinoma including those of the breast, endometrium ovary, and other sites. Cells from four established human tumor cell lines (three breast and one endometrial) were also included in this study. The results showed that addition of EGF at a concentration of 50 ng/ml resulted in a 50% higher cloning efficiency in soft agar in 40% of the samples of fresh human tumors. When cells from tumor cell lines were plated in semi-solid medium containing EGF, the number of colonies formed was at least twice as high as controls. Cells from fresh tumor biopsies were assayed for EGF receptors to determine whether the correlation between the proliferative response in EGF-supplemented semi-solid medium as compared to control could be related to the number of EGF receptors present on the cells. Specific receptors for EGF were detected by using radioiodinated EGF in early passage cell cultures from some of the tumors tested for clonogenicity. The number of receptors ranged from 0.3 to 3.27 X 10(5) per cell. Cells from two melanoma specimens possessed less than 0.3 X 10(3) EGF receptors per cell. We found no correlation between the number of EGF receptor on a cell surface and the mitogenic effect of EGF on the same tumor cells grown in semi-solid medium. PMID- 6298121 TI - Characterization of stable spontaneous metastatic variant lines of RSV transformed mouse fibroblasts. AB - The correlation between metastatic potential and a series of biological properties was investigated in two mouse fibrosarcoma lines (SR-BALB and B77 3T3), transformed by different strains of Rous sarcoma virus (RSV). In the absence of selective pressure the metastatic potential was different in the two lines. The SR-BALB sarcoma did produce both spontaneous metastases from s.c. site and i.v. lung colonization with a high incidence (respectively in 60% and 80% of treated animals). Conversely, the metastatic incidence of the B77-3T3 sarcoma was much lower. Differences in lung implantation between the two lines turned out to be even greater when the number of colonies growing in the lung was evaluated. Organ distribution of cells after i.v. injection, tumorigenicity, growth rate in vivo and in vitro, plating efficiency in liquid medium and cloning efficiency in semi-solid agar medium were evaluated in the two lines. A strict correlation was found only between the metastatic potential and the capability of growth in 0.6% ("hard") agar. Such a correlation was supported by the isolation in "hard" agar of highly metastasizing subclones of the low-metastasizing B77-3T3 line. PMID- 6298122 TI - Effect of elicitation on peritoneal macrophage subpopulations: size distributions, ectoenzyme phenotypes and antitumor activity. AB - Resident (R), Brewer's thioglycollate broth (TG) and C. parvum (CP)-elicited murine peritoneal cell populations were separated into subpopulations by centrifugation on discontinuous Ficoll gradients consisting of layers of 4, 6, 8 and 10% Ficoll. The resulting subpopulations were shown to be distinct on the basis of cell size, ectoenzyme phenotypes and antitumor activity. The cell size distributions were analyzed by means of a Coulter channelyzer and software developed for a computer. The 4-6% Ficoll interface fraction comprised the smallest macrophages, with most cells in this subpopulation appearing to range in cell volume from 150-275 microns 3. The largest sized macrophages were found in the 10%-pellet fraction, with most cells appearing to range in cell volume from 300-600 microns 3. The ectoenzyme phenotypes of the R, TG and CP unseparated macrophage populations were significantly different. Moreover, the ectoenzyme phenotypes of the smallest, (4-6% Ficoll interface) subpopulation in the R and CP macrophages differed from the other subpopulations. Reduction in alkaline phosphodiesterase 1 (APD-1) ectoenzyme activity (as compared with unseparated R macrophages) appeared to be a marker for acquisition of antitumor activity. The small CP macrophages (4-6% Ficoll interface) showed no antitumor activity while the unseparated CP macrophages and all other CP macrophages subpopulations exhibited antitumor activity. The CP macrophage unseparated population and the subpopulations with antitumor activity expressed reduced ADP-1 activity. Conversely, the R or TG macrophage unseparated populations and their subpopulations, along with the CP macrophage 4-6% subpopulation, lacked antitumor activity and failed to show a change in ADP-1 ectoenzyme activity. PMID- 6298123 TI - DNA virus-transformed hamster cell--host effector cell interactions: level of resistance to cytolysis correlated with tumorigenicity. AB - Spontaneously cytolytic hamster spleen cells and BCG-activated hamster macrophages were used to examine susceptibilities to nonspecific effector cell induced lysis among 13 DNA virus-transformed hamster cell lines exhibiting four different tumorigenic phenotypes. Hamster cells transformed by adenovirus type 12 (an oncogenic adenovirus serotype) or simian virus 40 (an oncogenic papovavirus) readily induced tumors in immunocompetent syngeneic hamsters and were relatively resistant to spleen-cell-induced lysis compared to cells transformed by adenovirus type 2 (a non-oncogenic adenovirus serotype) which induced tumors only in immunoincompetent hosts. Simian virus 40-transformed cells, which possess the unusual property of efficient tumor induction in allogeneic hosts, were uniquely resistant to lysis by activated macrophages. These differential patterns of susceptibility to cytolysis suggest an association between the level of transformed cell resistance to lysis by nonspecific host effector cells and the oncogenicity of the transforming virus. Furthermore, these data suggest that tumor-cell properties, other than those commonly associated with neoplastic transformation, determine the level of susceptibility or resistance to host effector cell mechanisms. PMID- 6298124 TI - Characterization and activation of cyclic adenosine 3':5'-monophosphate-dependent protein kinase in C1300 murine neuroblastoma clones growing in vivo. AB - Cyclic adenosine 3':5'-monophosphate (cAMP)-dependent protein kinases from three clones of C1300 neuroblastoma, established as growing tumors in A/jax mice, were identified and characterized. Mean (+/- SD) intratumor concentrations of cAMP ranged from 5.0 +/- 2.4 to 6.8 +/- 1.9 pmol/mg protein; cytosolic protein kinase activity ratios, calculated from the -cAMP/+cAMP value, ranged from 0.12 +/- 0.02 to 0.18 +/- 0.02. The total amount of cytosolic cAMP-binding activity was highest in the NBA2 clone and nearly equal in N-18 and NBP2. By DEAE chromatography, two peaks of cAMP-binding activity eluting at ionic strengths of 0.09 and 0.15 M of NaCl were resolved from each clone. The latter peak was associated with catalytic activity (type II cAMP-dependent protein kinase), whereas the former was not (free type I cAMP-binding protein). Photoaffinity labeling with 8-azido (N3) [32P]cAMP, followed by sodium-dodecyl-sulfate:polyacrylamide gel electrophoresis, resolved four major cAMP-binding proteins with molecular weights ranging from 39,000 to 56,000 in each clone. The mol. wt 47,000 protein was judged to be a free regulatory subunit of type I kinase, and the mol. wt 39,000 protein a cleavage product. The mol. wt 54,000 and 56,000 proteins were both endogenously phosphorylated and both had a lower affinity for 8-N3-[32P]cAMP, characteristic of type II kinase. Selected properties of the cAMP-binding proteins (kinetic and autophosphorylating features, response to tryptic hydrolysis, specificity for cAMP and temperature sensitivity) did not differ appreciably among clones. Despite initially low intratumor concentrations of cAMP, it was possible to activate the protein kinase system by simultaneous injection of N6, O2'-dibutyryl cAMP and papaverine. This indicates that mouse C1300 neuroblastoma can be used profitably to study cAMP-induced neuroblast differentiation. PMID- 6298125 TI - Relationship between the amounts of EBV-DNA and EBNA per cell, clonability and tumorigenicity in two ebv-negative lymphoma lines and their EBV-converted sublines. AB - The effect of EBV-conversion of two EBV-negative lymphoma lines (Ramos and BJAB) on agarose clonability and tumorigenicity in nude mice was explored. The cloning frequency was increased in all 9 sublines investigated, between 1.1 and 4.9 times compared to the original "parental" lines. Tumorigenicity was increased in one out of 2 BJAB-derived, EBV-positive lines and in 3 of 6 Ramos-derived lines, while it was decreased in 3 others. A strong positive correlation between the number of genomes/cell and the amount of EBNA/cell was detected, and in 7 of 9 converted lines the cloning frequency also correlated to the number of EBV genomes. On the other hand, no relation was established between the number of EBV genomes/cell, the amount of EBNA/cell and tumorigenicity. PMID- 6298126 TI - Epstein-Barr virus-specific antibodies in patients with adult T-cell leukemia (ATL) and healthy ATL virus-carriers. AB - Antibodies to Epstein-Barr virus (EBV)-capsid antigen (VCA), early antigen (EA) and EBV-associated nuclear antigen (EBNA) in the sera of 103 patients with adult T-cell leukemia (ATL) and the sera of 243 age- and sex-matched healthy adults, namely 99 anti-ATLA-positive (antibodies to ATL virus-associated antigen) and 144 anti-ATLA-negative individuals, were determined by indirect immunofluorescence. The anti-VCA titers in the sera from ATL patients were within the range observed in healthy controls. Anti-EA was found in 27% of the sera from ATL patients, but in only 8% of the sera from anti-ATLA-negative healthy controls. Antibodies to EBNA, which were present in almost all healthy adults, were not found in 30% of the sera from ATL patients. Of the sera of anti-ATLA-positive healthy adult donors 11% were negative for EBNA antibody. These results suggest that functional impairment of the T-cell system in most ATL patients, and also to a lesser extent in anti-ATLA-positive adults who might be healthy ATLV-carriers, may cause an unusual immune response of antibodies to EBV-associated antigens. PMID- 6298127 TI - Effect of hormones on the expression of proviral genes Mtv-2 and Mtv-3 in mouse mammary gland. AB - We have investigated the expression of the Mtv-2 and Mtv-3 proviral genes in mouse mammary glands by examining the effect of hormones on levels of mammary tumor virus (MTV) proteins p27 (gag) and gp52 (env) in mouse mammary explants. We also investigated the effect of the hormones on DNA synthesis in the explants. The mammary glands were derived from inbred GR and 020 mice, and from the respective congenic mouse strains GR/Mtv-2- and 020/Mtv-2+. The addition of insulin to the culture medium caused increases in p27 and gp52 levels in GR and 020/Mtv-2+ glands; a further increase in the viral proteins was obtained by also adding dexamethasone. Prolactin in combination with progesterone enhanced p27 and gp52 levels, but to a lesser extent than did dexamethasone. Dexamethasone caused a slight but significant increase in p27 protein in mammary explants from GR/Mtv 2- mice. Our data indicate that the Mtv-2 locus and the Mtv-3 locus in mouse mammary gland are under separate glucocorticoid control, and that Mtv-2 expression is also stimulated by the prolactin and progesterone combination. Whereas dexamethasone enhances MTV protein levels in mouse mammary explants, it inhibits DNA synthesis in the explants. PMID- 6298128 TI - The presence of p53 transformation-related protein in Ab-MuLV transformed cells is required for their development into lethal tumors in mice. AB - p53, a cellular-encoded protein, is synthesized at elevated levels in a wide range of tumor cells. Ab-MuLV-transformed cells expressing both the viral-encoded p120 oncogene and the cellular-encoded p53 display a lethal tumor phenotype in syngeneic mice. L12 is an exceptional Ab-MuLV-transformed cell line that expresses the p120 oncogene and lacks the p53 cellular protein. Injection of L12 cells into syngeneic mice is followed by the development of local tumors that are subsequently rejected. Prolonged treatment of L12 cells with TPA, a tumor cell promoter, gave rise to L12T cells that synthesize the p53 protein and exhibit a lethal tumor phenotype. Comparison of one-dimensional proteolytic partial peptide map of p53 obtained from L12T to that obtained from other Ab-MuLV-transformed cell lines confirmed their identity. These results suggest a correlation between the cellular expression of p53 in Ab-MuLV-transformed cells and their capacity to develop into lethal tumors in syngeneic mice. PMID- 6298129 TI - Genetic control of the organ specificity of lymphoproliferative disease virus (LPDV) of turkeys. AB - In a previous study based on the kinetics of virus replication and tumor formation (Gazit et al., 1982), it was shown that the organotropism of lymphoproliferative disease virus (LPDV) is confined to lymphoid tissues. The present paper demonstrates that this organ specificity is controlled at the level of infection and integration, that is, the lymphoid organs, which are the only organs sustaining virus replication, and also the only organs in whose cells integrated LPDV proviruses are detectable. At the same time, the efficiency of virus replication within the various target organs is regulated both at the level of infection and integration and at the level of viral gene transcription. PMID- 6298130 TI - Collagenase activity in rabbit carcinoma: cell source and cell interactions. AB - Fibroblast-like cells (F-cells) and epithelial-like (E-cells) derived from cultures of rabbit VX-2 carcinoma released collagenase in both active and latent forms in serum-free medium at a level higher than that of normal rabbit fibroblast cultures. The enhanced capacity of the F-cells to release the enzyme, however, continued only for a few passages and then decreased significantly to a low level similar to that of the normal fibroblasts. The enzyme-release by the E cells continued for a few more passages at a relatively moderate level, higher than that of normal fibroblasts. The release of collagenase in cultures of F cells was enhanced by the presence of E-cells in mixed cultures as well as by medium conditioned by the E-cells type. Addition of cytochalasin B at 2 micrograms/ml did not significantly effect the enzyme activity released in the cultures. Serum from tumor-bearing rabbits appeared to stimulate the release of enzyme activity in cultures of either cell type. PMID- 6298131 TI - Lymphocyte thyroid hormone receptors in obesity. AB - Triiodothyronine (T3)-receptor characteristics of isolated circulating human mononuclear cells have been studied in a group of obese patients who claimed to be unable to lose weight on conventional 4.2 MJ (1000 kcal) diets. The cells of the obese patients exhibited a lower receptor capacity than those of a control group of non-obese subjects but the difference was not significant. There was a significant fall (P less than 0.01) in receptor capacity in the obese patients after 12 weeks in a 1.34 MJ (320 kcal) per d formula diet and this provides a further mechanism whereby a fall in metabolic rate takes place in response to severe dietary restriction. Some patients who also received T3 (60 micrograms/d) in addition to the formula diet showed a further fall in the receptor numbers. These findings may partly account for the previously reported resistance to thyroid hormones in obesity. PMID- 6298132 TI - Enzymes in the exsheathing fluid of nematodes and their biological significance. PMID- 6298133 TI - Nitroxyl-induced T1 relaxation rates as a probe of conformation in peptide hormones. AB - Proton spin-lattice relaxation rates in the N-H region of the n.m.r. spectra of aqueous LHRH and angiotensin II were measured in the presence of varying concentrations of 2,2,6,6-tetramethylpiperidinoxyl. At peptide concentrations of 3-7 mM and nitroxyl concentrations up to three times the peptide concentration, the relaxation rate is linearly dependent on nitroxyl concentration. Second order rate constants for nitroxyl induced relaxation in water are in the 200-1000s-1 M 1 range, and are dependent more on conformational factors than on side chain bulk. Exclusion of the radical from the hydration sphere of imidazolium ion appears to occur. The measurements for LHRH agree with earlier observations of the effect of the nitroxyl on linewidths. A two- to three-fold decrease in sensitivity of the amide protons of at least three of the residues in angiotensin II when the His6 imidazole is protonated indicates a conformational transition related to this ionization. PMID- 6298134 TI - Yields of hydroxyl radical and reducing species in tritiated water based on the radiolysis of tetranitromethane. AB - Yields of the nitroform anion produced in tetranitromethane aqueous solutions including tritiated water with and without isopropanol were measured. Yields of .OH and reducing species (e-aq + .OH) were calculated from the yields of the nitroform anion. The G values obtained for them were 3.8 and 2.5, respectively. The former was higher and the latter was lower than those for 60Co gamma-rays. Their beta/gamma ratio was 1.36 and 0.76, respectively. The beta/gamma ratio, 1.36, for .OH may be related to r.b.e. values higher than the one reported for 3H beta-rays. PMID- 6298136 TI - Effect of ionizing radiation on the transport of spin-labelled compounds across the porcine erythrocyte membrane. II. PMID- 6298135 TI - Mechanism of ultraviolet light induced catabolite repression of L-arabinose isomerase. PMID- 6298137 TI - Clostridium perfringens meningitis. AB - In this report, primary Clostridium perfringens meningitis, a rare condition, is described in a 62-year-old man who developed fatal clostridial meningitis following surgical evacuation of a subdural hematoma. This is the third case report of clostridial meningitis as a complication of craniotomy. Clostridial infection should be considered in any patient with meningitis with a history of surgical intervention, or head injury, even if only slight. PMID- 6298138 TI - Corneal glycosaminoglycan synthesis in long-term organ culture. PMID- 6298139 TI - Herpes simplex virus recovery in neural tissues after ocular HSV shedding induced by epinephrine iontophoresis to the rabbit cornea. AB - Ocular HSV-1 shedding from latently infected rabbits was induced by iontophoresis of 0.01% epinephrine into the eye. Anodal Iontophoresis of epinephrine was performed at 0.8 mAmps for 8 min once a day for 3 consecutive days. Shedding was determined by the presence of HSV-1 in the preocular tear film obtained via eye swabs. Bilateral epinephrine iontophoresis performed on selected days during 220 280 days after inoculation resulted in HSV-1 shedding in 75% of the eyes (30/40) and 100% of the rabbits (20/20). Following the induction of ocular HSV-1 shedding, rabbits were killed and selected neural tissues were homogenized. Cell free preparations were assayed for the presence of infectious virions using primary rabbit kidney cell monolayers. When the tissues were homogenized immediately after death, virus was detected in only one neural tissue, the trigeminal ganglia. However, when the tissues were incubated in vitro for 18-24 hours prior to the homogenization, infectious HSV-1 was recovered from homogenates of the trigeminal ganglion, superior cervical ganglion, the ophthalmic branch of the trigeminal nerve, and the root entry zone of the trigeminal nerve. A relationship was noted between the time of the last ocular shedding and recovery of infectious HSV from the tissue homogenates. Furthermore, a positive correlation in 11 eyes between the recovery of HSV-1 from the perocular tear film and HSV-1 recovery from one or more corresponding neural tissues was found. These results suggested that epinephrine iontophoresis to the cornea triggered an "alteration" in the state of the virus in the neural tissues of the latently infected rabbits and that the change can be related to the induced ocular shedding. PMID- 6298140 TI - Enzymatic disaggregation of the infected rat cornea. AB - Analysis of cell populations in the cornea may be performed rapidly and accurately employing the technique of enzymatic disaggregation. To illustrate this method normal rat corneas and corneas infected 24 and 48 hours previously with Staphylococcus aureus were disaggregated in a solution containing pancreatin and collagenase. The cells released were counted and identified morphologically. These results were compared to cell counts made from histologic sections. Over 95% of the corneal cells were viable after the disaggregation and leukocytes obtained from the infected corneas retained their phagocytic capacity. This approach allows sensitive analysis of cell populations in a wide range of corneal conditions, including infection and allograft rejection. PMID- 6298141 TI - Junin virus infection in genetically athymic mice. AB - The progression of Junin virus infection was studied in congenitally athymic mice. Immunocompetent littermates were used as infected controls. As expected, the latter developed lethal encephalitis, with viremia and considerable viral replication in the brain. The mortality rate was almost 100%; the few surviving controls exhibited high serum neutralizing antibody levels and a total absence of virus in blood and brain. In contrast, nude mice did not contract the disease; all survived with persistent viremia and virus in brain, but no serum neutralizing antibodies were detected. These results confirm previous research on thymectomized mice and those treated with anti-lymphocyte serum and tend to support the important role of cellular immunity in the pathogenesis of this viral disease. PMID- 6298142 TI - Effect of phosphonoformate on symptomatic genital herpes simplex virus type 2 infection of guinea pigs. AB - Using a guinea pig model of genital herpes simplex virus (HSV) type 2 infection, phosphonoformate treatment initiated after the onset of symptoms had a therapeutic effect on the outcome of disease. Severity of disease, viral shedding, and HSV-induced cellular changes in vaginal cytology were significantly reduced in drug-treated animals as compared to sham-treated animals, although virus was readily detected in the nervous system of animals from both groups. The percentage of animals harboring latent virus in their dorsal root ganglia was approximately the same in both drug-treated and sham-treated animals. No drug toxicity was found. PMID- 6298143 TI - Cytogenetic effect of two strains of Junin virus in the guinea pig. AB - The cytogenetic effect of two strains of Junin virus on bone marrow chromosomes of the guinea pig was studied. Animals infected with the attenuated strain XJ-Cl3 showed no differences from control animals. Guinea pigs inoculated with the pathogenic strain XJ exhibited a significant increase of abnormal cells, chromatid breaks, and chromosome fragments. The clastogenic ability of the XJ strain is similar to the reported effect of other viruses, while the XJ-Cl3 strain does not appear to be clastogenic. PMID- 6298144 TI - Epstein-Barr virus carried by Raji cells: a mutant in early functions? AB - Immunoprecipitation was used to study Epstein-Barr virus-specific polypeptide synthesis in Raji cells after chemical induction of the viral cycle. The results indicate that Raji cells fail to synthesize a few early and both late polypeptides. Comparison with P3HR-1 virus superinfection experiments at different MOIs indicates that some of these early polypeptides are required for virus DNA replication. The results are compatible with what is known about the complexity of viral DNA in Raji cells. PMID- 6298145 TI - Functional complementation between Epstein-Barr virus and herpesvirus papio. AB - The possibility of complementation between herpesvirus papio (HVP) and Epstein Barr virus (EBV) was investigated. Strain 594S-F9 of HVP, unlike strain P3HR-1 of EBV, is not capable of inducing virus antigen synthesis in the EBV genome carrying, nonproducer lymphoma cell line Raji. The effects of dual infection with these viruses were studied. With untreated viruses, the percentage of cells positive for viral antigens was equal to or slightly less than that in cultures infected with P3HR-1 virus only. However, if UV-irradiated P3HR-1 virus was employed in the dual infection, the relative number of virus antigen-positive cells was enhanced over cultures infected with P3HR-1 virus alone. These results suggest functional complementation between EBV and HVP. PMID- 6298146 TI - Transformation in the presence of dimethyl sulfoxide facilitates recovery of Epstein-Barr virus. AB - Human umbilical cord blood lymphocytes were immortalized by infection with Epstein-Barr virus (EBV) in the presence of various concentrations of dimethyl sulfoxide (DMSO). 13 continuous lymphoblastoid cell lines were established. DMSO did not affect the efficiency of transformation and establishment of these lines. After a period of 2 months in culture, the lines were tested for EBV production by lethal irradiation (6,000 rad) from a 60Co source and subsequent cocultivation with primary umbilical cord lymphocytes. Cell lines transformed in the presence of 2% DMSO yielded transforming activity in 46.6% of test cultures, compared with 9.5% for lines established without DMSO in the medium. These findings imply that a permanent change in the host/virus relationship resulted from a brief exposure (48 h) to DMSO at the time of transformation. PMID- 6298148 TI - Comments on the transverse shielding of proton accelerators. PMID- 6298147 TI - Herd immunity to poliovirus in Dundee. PMID- 6298149 TI - Measurement of charged and unattached fractions of radon and thoron daughters in two Canadian uranium mines. PMID- 6298151 TI - Determination of vitamin D and retinoid activity in eggs by HPLC. AB - Reverse phase high performance liquid chromatography (HLPC) was used to estimate the vitamin D3 content of battery and free range eggs. At the same time another HPLC technique was used to estimate the retinol activity in the battery eggs derived from the separate determination of all-trans retinol, 13-cis retinol and retinaldehyde concentrations. There was some variation in both the vitamin D3 and retinol activity in eggs over the laying year but no significance could be drawn from this. Vitamin D3 levels in 12 monthly samples each of 120 battery eggs averaged 1.2 micrograms/100 g edible portion, while the levels in each of four quarterly samples of 36 free range eggs from the London and Newcastle areas averaged 1.4 and 0.8 micrograms/100 g respectively. These differences between battery and free range eggs were not statistically significant. The average retinol activity in four samples of 120 battery eggs was calculated to be 190 micrograms/100 g, and the components of this activity were identified as 132 micrograms all-trans retinol and 21 micrograms of retinaldehyde, as well as 53 micrograms 13-cis retinol which has not previously been reported in hens' eggs. PMID- 6298150 TI - Clinical aspects of drug intoxication: opioids and opiates. PMID- 6298153 TI - Cytochemical evidence of the origin of the dense tubular system in the mouse platelet. PMID- 6298152 TI - Diet and coronary heart disease. AB - There is a moderately good correlation between dietary saturated fatty acids and coronary heart disease (CHD) when populations in different parts of the world are compared but not within the same cultural community or for individuals. Total energy, essential fatty acids (EFA), dietary fibre, alcohol and salt also contribute to the relationship of diet to CHD. Saturated fatty acids exert their pathogenic role mostly through altering the homoeostasis of lipoprotein metabolism, leading to an increase in cholesterol-rich low-density lipoproteins, and influencing adversely the balance between the accumulation in and clearance of cholesterol esters from the arterial wall. Polyunsaturated fatty acids (PUFA) alter lipoprotein metabolism directly by decreasing synthesis and increasing catabolism and excretion and indirectly by being substitutes for saturated fatty acids, which are therefore consumed in smaller quantities. PUFA have an important additional role, because of their EFA content. A deficiency of the EFA, linoleic and arachidonic acids, relative to the saturated fatty acid intake, can be correlated with CHD mortality. The pathogenic pathways of a relative EFA deficiency leading to CHD are as likely to be related to intravascular coagulation and myocardial metabolism as to lipoprotein metabolism. EFA requirements may not be met in communities having a high intake of saturated fatty acids if there is also a dietary deficiency of antioxidants particularly vitamin E. Two large primary prevention trials of diets enriched with PUFA showed, under institutional circumstances, that it is possible to reduce serum cholesterol by 10-15 per cent. There was in each a reduction in the total incidence of cardiovascular diseases, and the greatest effect was on the incidence of non-fatal myocardial infarction. In both trials there was an increase in non-cardiovascular mortality. The significance of this finding is evaluated in conjunction with a similar finding derived from the only long-term primary prevention trial using a drug. The possibility that gradual depletion of membrane cholesterol over many years might have an adverse effect on the function of ageing cells cannot be dismissed. In communities where there is a high incidence of CHD, the aim should be to reduce plasma cholesterol concentrations to the region of 210 mg/dl. The evidence that it is necessary, practicable or even desirable to reduce plasma cholesterol to lower levels in order to reduce CHD incidence further is not impressive. To achieve this aim by dietary measures there should be a reduction of energy from fats to a level of 35 per cent or below and reduction of the contributions from saturated fats to about 10 per cent. In addition, there is a sound basis for recommending reduction of total energy intake to that actually needed, an increase in dietary cereal fibre and a reduction in alcohol. PMID- 6298154 TI - [Care of patients during and after radiotherapy of malignant otorhinolaryngologic tumors]. PMID- 6298155 TI - The particles compared. AB - Figure 5 represents an updated attempt to represent the dose distribution and high LET advantages of the various heavy particles, summarizing the conclusions discussed above. Neutrons are the cheapest form of high LET radiation. Their biological properties are excelled only by argon ions, but of course, they yield quite ordinary dose distributions. Protons give the best physical dose distribution and are by far the cheapest charged particles. Their biological properties, however, are similar to gamma-rays. Pions are comparable to carbon and neon ions in dose effect distribution, but are not as good because of dose/rate limitations and a less sharp edge to the beam. Two principal questions remain: 1. Are super dose distributions worth the substantial cost? 2. Do hypoxic cells limit radiocurability? These questions can only be answered in clinical trials and cannot even be addressed in the radiobiology laboratory. PMID- 6298156 TI - Fast neutron beam radiation therapy in the United States. AB - Following a 30-year hiatus after Dr. Stone's work in the 1930's and 1940's, clinical trials with fast neutrons were restarted in the United States in 1972. Approximately 2500 patients have been treated with neutrons since that time. Three hundred and seven patients with squamous cell carcinomas of the head and neck were entered on an RTOG-coordinated randomized study comparing standard photon irradiation with mixed beam radiation therapy. No significant differences were noted in the local control, survival or complication rates. One hundred and sixty patients were entered on a randomized glioblastoma study. Although there were no significant differences in median survival, autopsy results showed greater tumor effect on the neutron-treated tumors. Twenty-six patients were treated for transitional cell carcinomas of the bladder with either preoperative mixed beam irradiation or mixed beam irradiation alone. Both the local control rates and survival rates compare favorably with photon radiation therapy. The future of fast neutron beam radiation therapy in the United States is discussed. PMID- 6298157 TI - Japanese experience with clinical trials of fast neutrons. PMID- 6298158 TI - Pion radiotherapy at LAMPF. AB - Clinical investigations of pi meson radiotherapy were conducted by the Cancer Research and Treatment Center of the University of New Mexico and the Los Alamos National Laboratory from 1974 until 1982. Two hundred and thirty patients have been treated for a variety of locally advanced primary and metastatic neoplasms. One hundred and ninety-six patients have been followed for a minimum of 18 months. Crude survival data range from 11% for unresectable pancreatic carcinoma to 82% for Stages C and D1 adenocarcinoma of the prostate. Acute tolerance of normal tissues is approximately 4500 pion rad in 36 fractions over 7 weeks. Severe chronic reactions have appeared with increasing frequency after doses in excess of 4000 pion rad. PMID- 6298159 TI - Pions, Vancouver. AB - Clinical treatments at TRIUMF started in November, 1979. Ten patients with malignant subcutaneous nodules had 14 lesions treated with pions and 37 other nodules treated with 280 kV X rays. Three different fractionation regimens were used with X ray doses spanning the expected RBE range of pions. The RBE for pions for acute skin reaction for 10 fractions had a mean value about 1.5, while for 3 fractions it was 1.3 maximum. No dissociation of acute and late skin effects was seen with follow-up to 27 months after treatment. Phase 1-2 studies of Pion-Boost Therapy for patients with glioblastoma multiforme will begin in May, 1982. These will be followed in August with treatments of advanced pelvic malignancies using pions only. The existing beam line at TRIUMF will be upgraded and commitments have been given to go to higher beam currents. As a result, the dose rate should increase by a factor of at least two, allowing treatment of clinically relevant volumes in acceptable times by 1983-1984. PMID- 6298160 TI - Evaluation of the clinical applicability of proton beams in definitive fractionated radiation therapy. AB - We report on the treatment of 317 patients treated either wholly or in part with proton beams at the Harvard Cyclotron Laboratory. These include: 130 patients treated for definitive radiation therapy of choroidal melanoma; 17 patients treated for tumors of the base of skull, cervical spine and cranium, which abut structures of the central nervous system (CNS); 23 patients treated for sarcomas of soft tissue and bone; 65 patients treated for carcinoma of the prostate; 14 patients treated for carcinoma of the rectum and anus; and 23 patients treated for squamous carcinoma of the oral cavity and oro-pharynx. Data on causes of failure and morbidity of treatment are presented. Overall the results are judged to be extremely encouraging. In particular, the treatment of the choroidal melanomas and sarcomas abutting CNS structures have clear clinical value, and the treatment of prostatic tumors and tumors of the head and neck are thought to be promising. PMID- 6298161 TI - Procedures for the elimination of pseudorabies virus from herds of swine. PMID- 6298162 TI - Treatment of feline cryptococcosis with ketoconazole. PMID- 6298163 TI - Treatment of canine blastomycosis with ketoconazole. AB - Four dogs with blastomycosis were treated with ketoconazole, and good results were obtained in 3 of the 4 cases. Dogs that responded to treatment had manifested a variety of the clinical forms of blastomycosis (osseous, pulmonary, lymphatic, and ocular), and all lesions in the dogs abated. There were no clinical signs or clinicopathologic evidence of adverse drug reaction or toxicosis. The results of the study suggested that ketoconazole is effective for the treatment of canine blastomycosis. PMID- 6298164 TI - Insulin therapy in cats with diabetes mellitus. AB - Thirteen cats with diabetes mellitus were evaluated. Clinical signs included polydipsia, polyuria, polyphagia, lethargy, and weight loss. Results of physical examination included obesity, hepatomegaly, mild seborrhea sicca, muscle wasting, and dehydration. One cat walked plantigrade and was suspected of having a diabetic neuropathy. Persistent hyperglycemia, glucosuria, high liver enzyme activities, hypercholesterolemia, hyperproteinemia, and low electrolyte concentrations were the common laboratory findings. In 3 cats diabetes mellitus developed after megestrol acetate therapy; 2 of these cats required only temporary insulin treatment. In a 3rd cat, which had no history of receiving diabetogenic drug therapy, remission of diabetes mellitus also was observed. Serum insulin and plasma glucose concentrations were determined in 6 cats after administration of an intermediate-acting insulin (isophane insulin) and in 3 cats after administration of a long-acting insulin (protamine zinc insulin). The insulin concentration peaked 2 to 6 hours after the injection of intermediate acting insulin and 6 to 12 hours after the injection of long-acting insulin. The lowest glucose concentration was recorded 4 to 8 hours after injection of intermediate-acting insulin, and 6 to 12 hours after injection of long-acting insulin. It was concluded that, although insulin therapy must be adjusted to the individual, the diabetic cat usually requires twice-daily administration of isophane insulin; however, the protamine zinc insulin can be given once daily for satisfactory control. PMID- 6298165 TI - An inhibitor of (Na+, K+)-ATPase produced by Streptomyces pseudovenezuelae MF722 02; purification and properties. AB - A culture product of Streptomyces pseudovenezuelae MF722-02, with a molecular formula of C29H32N2O7, was isolated as yellow needles from culture broths and mycelia of the organism by means of a series of solvent extraction, column chromatography and crystallization. The antibiotic is active against some Gram positive bacteria, inhibits growth in vitro of cells of mouse leukemia L-1210, prolongs the life span of mice inoculated with the leukemia cells, enhances deoxycholate-induced hemolysis in vitro and inhibits (Na+, K+)-ATPase in vitro. PMID- 6298166 TI - Custom made C18 columns for the hydrophobic chromatography of cephalosporin C derivatives. AB - "Hydrophobic chromatography", which is a variation of reverse phase chromatography, is applicable to the analysis of cephalosporin C derivatives, especially in fermentation broths. Unfortunately, there are no commercial C18 columns which are entirely suitable for this class of compounds. For this reason C18 columns were prepared by an in-situ bonding technique and were optimally designed for cephalosporin C derivatives. Mono-, di- and trifunctional octadecyl bonding agents were used with 10 microns silica of both 60 A and 100 A pore diameter. The best results were obtained with the difunctional agent, methyloctadecyldichlorosilane, and 100 A silica. "Endcapping" of residual silanol groups with a trimethylsilylation agent was optional, since good results were obtained with both a plain C18 column and one that was "endcapped". PMID- 6298167 TI - 6-Acetylmethylenepenicillanic acid (Ro 15-1903), a potent beta-lactamase inhibitor. I. Inhibition of chromosomally and R-factor-mediated beta-lactamases. AB - 6-Acetylmethylenepenicillanic acid (Ro 15-1903) was seen to be a powerful inhibitor of various beta-lactamases. Nearly all of the chromosomally and R factor-mediated beta-lactamases which were studied were inhibited at much lower concentrations than required with clavulanic acid or sulbactam. Ro 15-1903 protected beta-lactamase-labile penicillins and cephalosporins from hydrolysis. The compound itself was stable against hydrolysis and inhibition of beta lactamase was irreversible. PMID- 6298168 TI - 6-Acetylmethylenepenicillanic acid (Ro 15-1903), a potent beta-lactamase inhibitor. II. Antibacterial properties. AB - The beta-lactamase inhibitor Ro 15-1903 showed low affinity for penicillin binding proteins (PBPs) of Escherichia coli. When used as a single compound, it displayed no substantial antibacterial activity but in combination with ampicillin, it was similar to clavulanic acid in conferring activity against ampicillin-resistant strains. Some synergy between Ro 15-1903 and piperacillin was found against high inocula of Pseudomonas aeruginosa. Ro 15-1903 markedly enhanced the activity of ceftriaxone against Bacteroides fragilis. In keeping with the in vitro findings, the combination Ro 15-1903 and ampicillin protected mice against systemic infections with beta-lactamase-producing strains of Staphylococcus aureus, Klebsiella pneumoniae and Proteus sp. but not against those with Enterobacter cloacae, Serratia marcescens, and E. coli producing either chromosomally mediated beta-lactamase of type I or plasmid-mediated beta lactamase of type TEM. PMID- 6298169 TI - Cefoxitin, N-formimidoyl thienamycin, clavulanic acid, and penicillanic acid sulfone as suicide inhibitors for different types of beta-lactamases produced by gram-negative bacteria. AB - Cefoxitin, N-formimidoyl thienamycin (MK0787), clavulanic acid, and penicillanic acid sulfone (CP45,899) were studied to determine their potency as suicide inhibitors of three very different kinds of beta-lactamases produced by Gram negative bacteria: type Ib penicillinase (TEM-2 type), a typical cephalosporinase (the enzyme of Proteus morganii), and a cephalosporinase with broad substrate specificity (the enzyme of Proteus vulgaris). All these beta-lactams were confirmed to be quite stable to the three beta-lactamases. The absolute values of the turnover numbers (kcat) for these enzyme-catalyzed hydrolyses were determined, the values ranged from 0.25 minute-1 to 660 minute-1. All the beta lactams studied, except cefoxitin, acted as suicide inhibitors of the typical cephalosporinase. Although cefoxitin did not exhibit such an effect, it was a powerful competitive inhibitor of this enzyme. Although the four beta-lactams acted as suicide inhibitors of the P. vulgaris cephalosporinase, the inactivated enzyme partially regained its activity after removing the effect of the free inhibitor. Type Ib penicillinase was also inactivated by the four beta-lactams, and the activity of the inactivated enzyme, except in the case of cefoxitin, was partially restored. The rate constants for enzyme inactivation or reactivation were calculated and are presented. The information obtained from this study suggests that the catalytic center of the P. vulgaris cephalosporinase is different not only from that of the penicillinase-type but also from that of the cephalosporinase-type beta-lactamases. PMID- 6298170 TI - Induced gonadotropin release in adrenocorticotropin-treated bulls and steers. AB - The effect of adrenocorticotropin hormone (ACTH) on plasma cortisol and on gonadotropin releasing hormone (GnRH)-induced release of luteinizing hormone (LH), follicle-stimulating hormone (FSH), and testosterone was determined in nine Holstein bulls and 12 Holstein steers. Treatments consisted of animals receiving either GnRH (200 micrograms, Group G), ACTH (.45 IU/kg BW, Group A) or a combination of ACTH followed 2 h later by GnRH (Group AG). Group G steers and bulls had elevated plasma LH and FSH within .5 h after GnRH injection and plasma testosterone was increased by 1 h after GnRH injection in bulls. In Group A, plasma cortisol was elevated by .5 h after ACTH injection in both steers and bulls, but plasma LH and FSH were unaffected. In Group A bulls, testosterone was reduced after ACTH injection. In Group AG, ACTH caused an immediate increase in plasma cortisol in both steers and bulls, but did not affect the increase in either plasma LH or FSH in response to GnRH in steers. In Group AG bulls, ACTH did not prevent an increase in either plasma LH, FSH or testosterone in response to GnRH compared with basal concentrations. However, magnitude of systemic FSH response was reduced compared with response in Group G bulls, but plasma LH and testosterone were not reduced. The results indicate that ACTH caused an increase in plasma cortisol, but did not adversely affect LH or FSH response to GnRH in steers and bulls. Further, while testosterone was decreased after ACTH alone, neither ACTH nor resulting increased plasma cortisol resulted in decreased testosterone production in the bull after GnRH stimulation. PMID- 6298171 TI - The pharmacokinetics of ketoconazole in severely immunocompromised patients. PMID- 6298173 TI - Early inhibition of human cytomegalovirus replication by novobiocin. PMID- 6298172 TI - In-vitro and in-vivo efficacy of ceftriaxone, moxalactam, and chloramphenicol against Haemophilus influenzae type b. PMID- 6298174 TI - Combined activity of ketoconazole and sulphamethoxazole against Candida albicans. PMID- 6298176 TI - Ceftriaxone therapy of group B streptococcal bacteraemia and meningitis in infant rats. AB - The efficacy of ceftriaxone against group B streptococci was studied in vitro and in vivo with an infant rat model of group B streptococci bacteraemia and meningitis. Twenty-four strains demonstrated minimal inhibitory concentrations of ceftriaxone of 0 . 05-0 . 1 mg/l and minimal bactericidal concentrations of 0 . 1 0 . 4 mg/l. Four strains were selected to induce bacteraemia and meningitis in infant rats by intraperitoneal inoculation. All 45 bacteraemic animals with or without meningitis that were treated with ceftriaxone 2 mg/kg/dose every eight hours for five doses survived, while all 12 control animals died (P less than 0 . 001). When recultured 54 h after the last dose of ceftriaxone, both CSF and blood remained sterile in all treated animals. These results indicate group B streptococci to be sensitive to ceftriazone in vitro and that, in the low dosage used, ceftriaxone effectively eradicates group B streptococcal bacteraemia and meningitis in infant rats. PMID- 6298175 TI - The effects of ketoconazole on cellular and humoral immune functions. AB - The effects of ketoconazole at varying concentrations on in-vitro neutrophil random migration, chemotaxis to autologous endotoxin--activated serum and the synthetic chemotactic tripeptide N-formyl-L-methionyl-L-leucyl-L-phenylalanine, phagocytosis and postphagocytic hexose monophosphate shunt activity and myeloperoxidase--mediated protein iodination were investigated. Neutrophil functions, in-vivo mitogen-induced lymphocyte transformation and levels of serum immunoglobulins and complement components were also assessed before and after ingestion of ketoconazole by six individuals. It was found that ketoconazole caused stimulation of neutrophil migration to leucoattractants 2 h after ingestion of a single dose of 400 mg ketoconazole. This stimulation was not sustained. The drug had no effect on the random migration of neutrophils. Likewise serum levels of the immunoglobulins IgG, IgM and IgA, total haemolytic complement and serum levels of the complement components C3 and C4 were unaffected. Ketoconazole in vitro had no effect on the neutrophil functions tested. Ingestion of ketoconazole caused a slight inhibition of lymphocyte mitogen-induced transformation. PMID- 6298177 TI - Identification of the uvrD gene product of Salmonella typhimurium LT2. AB - The product of the uvrD gene of Salmonella typhimurium LT2 and Escherichia coli K 12 is thought to play a role in both the correction of mismatched bases and the repair of DNA damage, since insertion mutations in the uvrD gene increase the spontaneous mutation frequency and make the cells more sensitive to killing by UV irradiation. To clone the uvrD gene of S. typhimurium, we first generated a uvrD specific probe by using DNA from an S. typhimurium uvrD421::Tn5 mutant. This probe was used to screen a lambda library of S. typhimurium DNA. Bacteriophage carrying intact uvrD+ genes were subsequently identified, and the uvrD+ gene was subcloned onto a low-copy-number vector. By using a combination of Tn1000 insertion mutagenesis and the maxicell technique, the product of the uvrD gene was shown to be a 75,000-dalton protein, and the relative direction of transcription of this protein was determined. Introduction of a low-copy-number plasmid carrying the S. typhimurium uvrD+ gene into uvrD insertion mutants of either S. typhimurium or E. coli restored the spontaneous mutation frequency and degree of UV sensitivity to the levels in the corresponding uvrD+ strains. PMID- 6298178 TI - Adenylate cyclase is required for chemotaxis to phosphotransferase system sugars by Escherichia coli. AB - We report that in Escherichia coli, chemotaxis to sugars transported by the phosphotransferase system is mediated by adenylate cyclase, the nucleotide cyclase linked to the phosphotransferase system. We conclude that adenylate cyclase is required in this chemotaxis pathway because mutations in the cyclase gene (cya) eliminate or impair the response to phosphotransferase system sugars, even though other components of the phosphotransferase system known to be required for the detection of these sugars are relatively unaffected by such mutations. Moreover, merely supplying the mutant bacteria with the products of this enzyme, cyclic AMP and cyclic GMP, does not restore the chemotactic response. Because a residual chemotactic response is observed in certain strains with residual cyclic GMP synthesis but no cyclic AMP synthesis, it appears that the guanylate cyclase activity rather than the adenylate cyclase activity of the enzyme may be required for chemotaxis to sugars transported by the phosphotransferase system. Mutations in the cyclic nucleotide phosphodiesterase gene, which increase the level of both cyclic AMP and cyclic GMP, also reduce chemotaxis to these sugars. Therefore, it appears that control of the level of a cyclic nucleotide is critical for the chemotactic response to phosphotransferase system sugars. PMID- 6298179 TI - Plasmid-borne sulfonamide resistance determinants studied by restriction enzyme analysis. AB - The relationship between sulfonamide resistance genes carried on different plasmids was investigated by restriction enzyme analysis and DNA-DNA hybridization. The results showed that sulfonamide resistance mediated by different plasmids is determined by the production of at least two different types of drug-resistant dihydropteroate synthase. Plasmids pGS01, pGS02, and R22259, found in bacteria isolated from patients in Swedish hospitals, contained identical sulfonamide resistance genes, which were also identical to those of plasmids R1, R100, R6, and R388. These latter plasmids, which have been well studied in different laboratories, were originally from clinical isolates from different parts of the world. Two other clinically isolated plasmids, pGS04 and pGS05, were shown to contain sulfonamide resistance determinants of a completely different type. PMID- 6298181 TI - Evidence for formation of superoxide and formate radicals in Methanobacterium formicicum. AB - Using spin labeling and spin trapping techniques in combination with electron paramagnetic resonance spectrometry, we have detected the formation of superoxide by whole cells of Methanobacterium formicicum under aerobic conditions in the presence and absence of sodium formate. Rates of superoxide generation have been estimated. The formation of additional free radical species, including formate, was observed. Production of these and other free radicals resulted in lipid peroxidation and concomitant cell damage. PMID- 6298180 TI - Unusual sulfonolipids are characteristic of the Cytophaga-Flexibacter group. AB - Capnocytophaga spp. contain a group of unusual sulfonolipids, called capnoids (W. Godchaux III and E. R. Leadbetter, J. Bacteriol. 144:592-602, 1980). One of these lipids, capnine, is 2-amino-3-hydroxy-15-methylhexadecane-1-sulfonic acid; the others are, apparently, N-acylated versions of capnine. The lipids were found, in amounts ranging from 2.5 to 16 mumol of capnoid sulfur per g of cells (wet weight), in two Cytophaga spp. and also in several closely related organisms: several Capnocytophaga spp., Sporocytophaga myxococcoides, two Flexibacter spp., and two Flavobacterium spp. With the exception of the flavobacteria, all of these bacteria have been shown to exhibit gliding motility. The two Flavobacterium spp. belong to a subset of that genus that shares many other characteristics with the cytophagas. Only the Capnocytophaga spp. contained large quantities of capnine as such; in all of the others, most (and possibly all) of the capnoids were present as N-acylcapnines. Capnoid-negative bacteria included some gliding organisms that may not be closely related to the cytophagas: two fruiting myxobacters, a gliding cyanobacterium (Plectonema sp.), Beggiatoa alba, Vitreoscilla stercoraria, Herpetosiphon aurantiacus, and Lysobacter enzymogenes. Nongliding bacteria representing nine genera were also tested, and all of these fell into the capnoid negative group. PMID- 6298182 TI - Plasmids of enterotoxigenic Escherichia coli H10407: evidence for two heat-stable enterotoxin genes and a conjugal transfer system. AB - Three species of plasmids, associated with virulence and conjugal transfer, were identified in a clinically isolated enterotoxigenic Escherichia coli strain, H10407 (serotype O78:H11). pCS1, a non-self-transmissible plasmid species with a molecular weight of 62 X 10(6) and a 47 mol% guanine-plus-cytosine content, specified colonization factor antigen I and heat-stable enterotoxin (ST) production, as reported by others previously. A second non-self-transmissible plasmid species, designated pJY11, with a molecular weight of 42 X 10(6) and a 51 mol% guanine-plus-cytosine content, specified ST and heat-labile enterotoxin production and manifested T5/T6 phage restriction. The third plasmid species, pTRA1, also had a molecular weight of 42 X 10(6) and had a guanine-plus-cytosine content of 51 mol%; this species was self-transmissible and promoted transfer of both pCS1 and pJY11 to other bacterial cells. pCS1 may have originated from species of bacteria with a lower guanine-plus-cytosine content than E. coli. Finally, although demonstrating some heterogeneity with each other, both STs encoded by pCS1 and pJY11 belonged to the STa group. PMID- 6298183 TI - Isolation and characterization of Tn5 insertion mutations in the lexA gene of Escherichia coli. AB - A Mu d(Ap lac)-generated fusion of lacZ to dinD, a gene induced by DNA damage, was used to isolate Tn5 insertion mutations that affect the regulation of the SOS responses. Three mutants were obtained that contained Tn5 insertions genetically linked to the lexA gene and had properties that suggested the mutants were deficient in lexA expression. The lexA protein has been shown to function as the repressor for genes involved in the SOS responses. By Southern blotting experiments, the three Tn5 insertions were physically mapped to distinct locations within the coding region of the lexA gene. The introduction of these mutations in six strains carrying lacZ fusions to different damage-inducible genes resulted in high expression of beta-galactosidase in all but one of the strains. In the dinF fusion strain, lacZ expression was reduced below that seen in a lexA+ background. Physical mapping studies of the dinF locus gave results consistent with the notion that dinF is part of the lexA transcription unit and that a lexA::Tn5 mutation has a polar effect on dinF expression. With certain din lac fusion strains, a correlation was seen between the amount of beta galactosidase production and the location of the particular Tn5 insertion within the lexA gene. PMID- 6298184 TI - Evolution of complex resistance transposons from an ancestral mercury transposon. AB - The molecular interrelationship of a transposon family which confers multiple antibiotic resistance and is assumed to have been generated from an ancestral mercury transposon was analyzed. Initially, the transposons Tn2613 (7.2 kilobases), encoding mercury resistance, and Tn2608 (13.5 kilobases), encoding mercury, streptomycin, and sulfonamide resistances, were isolated and their structures were analyzed. Next, the following transposons were compared with respect to their genetic and physical maps: Tn2613 and Tn501, encoding mercury resistance; Tn2608 and Tn21, encoding mercury, streptomycin, and sulfonamide resistance; Tn2607 and Tn4, encoding streptomycin, sulfonamide, and ampicillin resistance; and Tn2603, encoding mercury, streptomycin, sulfonamide, and ampicillin resistance. The results suggest that the transposons encoding multiple resistance were evolved from an ancestral mercury transposon. PMID- 6298185 TI - Two alanine racemase genes in Salmonella typhimurium that differ in structure and function. AB - Mutations were isolated in a previously undescribed Salmonella typhimurium gene encoding an alanine racemase essential for utilization of L-alanine as a source of carbon, energy, and nitrogen. This new locus, designated dadB, lies within one kilobase of the D-alanine dehydrogenase locus (dadA), which is also required for alanine catabolism. The dadA and dadB genes are coregulated. Mutants (including insertions) lacking the dadB alanine racemase do not require D-alanine for growth unless a mutation is introduced at a second locus, designated dal. Two genes specifying alanine racemase activity were cloned from S. typhimurium. The two cloned DNA sequences do not cross-hybridize with each other; one was shown to contain the dadB gene. PMID- 6298186 TI - Transposon-facilitated chromosome mobilization in Agrobacterium tumefaciens. AB - We improved chromosomal gene transfer in Agrobacterium tumefaciens strain 15955 by constructing donors containing homologous transposons on both the sex factor plasmid and chromosome. First, we constructed plasmid pDP35, a kanamycin sensitive derivative of R68.45. We then constructed derivatives of pDP35 that contained insertions of the kanamycin resistance transposon Tn5. By restriction endonuclease analysis, we identified two plasmids, pDP37 and pDP38, in which Tn5 was inserted in the same region of the plasmid but in opposite orientations. We also constructed isolates of A. tumefaciens containing an insertion of Tn5 in the chromosome. We transferred pDP37 or pDP38 into these chromosomal Tn5 strains and tested their ability to mobilize chromosomal markers to a series of auxotrophic recipients. Mobilization was observed at frequencies ranging from 10(-4) to 10( 7) recombinants per input donor for most markers tested. Both the plasmid and the chromosomal Tn5 elements were found to be required for mobilization at these higher frequencies. Donors were shown to transfer chromosomal markers in a polarized fashion. Recombinants coinherited unselected markers at frequencies of from 100 to 0.3 percent. The improved transfer frequencies and the observed polarity in chromosome transfer suggest that with this method we can genetically characterize A. tumefaciens chromosomal functions. PMID- 6298188 TI - chlC (nar) operon of Escherichia coli includes structural genes for alpha and beta subunits of nitrate reductase. AB - The synthesis of the alpha and beta subunits of nitrate reductase by 20 chlC::Tn5 insertion mutants of Escherichia coli was determined by immune precipitation of the subunits from fractions of cell extracts. Only two of the mutants produced either subunit in detectable amounts; these two accumulated the alpha subunit, but no beta subunit. In both cases the alpha subunit was present in the cytosolic fraction, in contrast to wild-type cells, in which both subunits are present mainly in the membrane fraction. EcoRI restriction fragments containing the Tn5 inserts from five of the mutants were cloned into pBR322. The insertions were localized on two contiguous EcoRI fragments spanning a 5.6-kilobase region that overlapped the contiguous ends of the two fragments. An insertion that permitted alpha subunit formation defined one end of the 5.6-kilobase region. The results indicated that the genes encoding the alpha and beta subunits of nitrate reductase were part of a chlC (nar) operon that is transcribed in the direction alpha leads to beta. PMID- 6298187 TI - cea-kil operon of the ColE1 plasmid. AB - We isolated a series of Tn5 transposon insertion mutants and chemically induced mutants with mutations in the region of the ColE1 plasmid that includes the cea (colicin) and imm (immunity) genes. Bacterial cells harboring each of the mutant plasmids were tested for their response to the colicin-inducing agent mitomycin C. All insertion mutations within the cea gene failed to bring about cell killing after mitomycin C treatment. A cea- amber mutation exerted a polar effect on killing by mitomycin C. Two insertions beyond the cea gene but within or near the imm gene also prevented the lethal response to mitomycin C. These findings suggest the presence in the ColE1 plasmid of an operon containing the cea and kil genes whose product is needed for mitomycin C-induced lethality. Bacteria carrying ColE1 plasmids with Tn5 inserted within the cea gene produced serologically cross-reacting fragments of the colicin E1 molecule, the lengths of which were proportional to the distance between the insertion and the promoter end of the cea gene. PMID- 6298189 TI - Comparison of Ti plasmids from three different biotypes of Agrobacterium tumefaciens isolated from grapevines. AB - Twenty-six plasmids from grapevine isolates of Agrobacterium tumefaciens were analyzed by SmaI fingerprinting and by hybridization of nick-translated DNA to DNA of another plasmid. These experiments established that octopine Ti plasmids are not highly conserved, although octopine Ti plasmids from biotype 1 A. tumefaciens strains appeared to be very similar. Octopine Ti plasmids from biotype 3 strains are more variable in terms of host range and SmaI fingerprints, but share extensive DNA homology. Fingerprints of nopaline Ti plasmids from strains of a given biotype resemble each other but not fingerprints of Ti plasmids from strains of the other two biotypes. The wide host range octopine Ti plasmid from the biotype 3 strain Ag86 shares more DNA homology with narrow host range Ti plasmids, nopaline Ti plasmids, and octopine catabolism plasmids than with the wide host range octopine Ti plasmid from biotype 1 strain 20/1. pTiAg86 does share homology with the portion of pTi20/1 integrated and expressed in plant tumor cells. Since all wide host range Ti plasmids studied contain these sequences, we suggest that natural selection for a wide host range resulted in the presence of the common sequences in distantly related plasmids. The lack of homology between this "common DNA" and limited host range Ti plasmids shows that the DNA sequences per se are not required for tumorigenesis. PMID- 6298190 TI - A sulfated polysaccharide produced by an Arthrobacter species. AB - A new sulfated polysaccharide was isolated from the culture supernatant of a strain of Arthrobacter sp. The polysaccharide purified with quaternary ammonium salts consists of D-galactose, D-glucose, sulfate, phosphorus, glucosamine, muramic acid, alanine, glutamic acid, glycine, and LL-diaminopimelic acid in a molar ratio of 56 : 9.0 : 68 : 6.4 : 2.0 : 1.1 : 2.1 : 1.0 : 1.2 : 1.2. The presence of the two amino sugars and four amino acids suggests that the polysaccharide, which is principally a galactan sulfate, contains small amounts of so-called peptidoglycan and that it is derived from the bacterial cell-wall polysaccharide. Gel filtration indicates the heterogeneity of the purified polysaccharide in its peptidoglycan content and molecular size. The molecular weight of its major portion was estimated to be 2.3 X 10(4) by gel filtration. The fractions GS-I and GS-II, and GS-4M and GS-5M, which were obtained by fractionation of the polysaccharide on Sephacryl S-200 and Dowex 1-X2 (Cl- form), respectively, gave almost the same chemical composition as the original polysaccharide, except in the peptidoglycan content, indicating that this polysaccharide is a series of complexes composed of essentially equal, sulfated polysaccharide chains and peptidoglycan fragments in their various ratios. The polysaccharide has [alpha]D -36 degrees and is composed predominantly of beta glycosidic linkages, as judged from its specific optical rotation (-37 degrees) and the infrared absorption (885 cm-1) of its desulfated material. It exhibits a potent antithrombin activity (ID50, 0.82 micrograms/ml). A possible partial structure of the polysaccharide is also discussed, based on the results of periodate oxidation, Smith degradation, and alkali treatment. PMID- 6298191 TI - A new aspect of a restriction endonuclease Tth111 I. It has a degenerated specificity (Tth111 I). AB - We previously reported that Thermus thermophilus 111 contained two restriction enzymes, Tth111 I and Tth111 II. The former does not cleave phi X174RFDNA and the latter does. We have now found another endonuclease activity able to cleave phi X174RFDNA in the cell extract of T. thermophilus 111. The protein with this activity was purified in a homogeneous state by chromatography on cellulose phosphate, heparin-Sepharose 4B and hydroxylapatite, successively. However, this endonuclease activity was always accompanied with Tth111 I activity during the purification procedure and the purified protein also showed a strong Tth111 I activity, suggesting that the Tth111 I activity and the phi X174RFDNA-cleaving activity reside in a single molecule. The phi X174RFDNA-cleaving activity was enhanced more strongly with Mn2+ than with Mg2+ and seemed to be attributable to a relaxed specificity of Tth111 I activity as seen in the cases of EcoRI* and BamHI* Thus we designated the phi X174RFDNA-cleaving activity Tth111 I*. The molecular weight of the protein with both Tth111 I and Tth111 I* activities was determined to be about 76,000 by gel filtration on a Sephadex G-100 column and 39,000 by SDS-polyacrylamide gel electrophoresis, suggesting the enzyme to be a dimer consisting of identical polypeptide chains. The phi X174RFDNA sequences surrounding Tth111 I* cuts were determined by the chain terminator method of Sanger et al. The results confirmed that Tth111 I* recognized a degenerated form of the Tth111 I recognition sequence, i.e., a sequence such that one of the specified nucleotides in the Tth111 I recognition sequence, 5'GACNNNGTC3', was substituted with N (N stands for any of A, G, C, and T), such as 5'NACNNNGTC3', 5'GACNNNNTC3', 5'GACNNNGNC, and so on (arrows indicate cleavage sites). PMID- 6298192 TI - Participation of calcium in the induction of phosphodiesterase by cyclic adenosine 3',5'-monophosphate in Dictyostelium discoideum. AB - The effects of divalent cations on the induction of phosphodiesterase [EC 3.1.4.17] by cyclic adenosine 3',5'-monophosphate (cyclic AMP) were studied in Dictyostelium discoideum. When cells were incubated with 1 mM ethylene glycol bis(beta-aminoethylether)-N,N,N',N'-tetraacetic acid (EGTA) in 20 mM Tris-HCl buffer, pH 7.5, for 2 h, the induction of cellular phosphodiesterase was inhibited by about 80%, and that of extracellular phosphodiesterase by about 65%. When cells were incubated with 1 mM EGTA for 1 h, 2 mM CaCl2 was added and the cells were further incubated for 1 h, the activities of cellular and extracellular phosphodiesterases were increased about 5 and 2.5 times, respectively, compared with those in the EGTA-inhibited cells. Although various other kinds of divalent cations were also studied, Ca2+ had the greatest effect on the induction. These results suggest that Ca2+ may participate in the induction of phosphodiesterase, and thus in the regulation of the development of the cellular slime mold. PMID- 6298193 TI - Respiration and photosynthesis in energy-transducing membranes of cyanobacteria. PMID- 6298194 TI - Tryptic digestion of the (Na + K)-ATPase is both sensitive to and modifies K+ interactions with the enzyme. AB - Tryptic digestion of the (Na + K)-ATPase in the presence of choline chloride or NaCl ("Na-type") and in the presence of KCl ("K-type") produced distinct patterns of peptide fragments and losses of catalytic activity. The K0.5 for K+ to shift digestion from the Na-type, and its sensitivity to dimethyl sulfoxide and Triton X-100, were consistent with K+ acting at sites on the cytoplasmic face of the enzyme through which the K-phosphatase reaction also is activated. Reagents favoring the E1 conformational states, oligomycin, Triton, and ATP, shifted the pattern toward the Na-type, whereas those favoring E2 states, dimethyl sulfoxide, MgCl2, and MnCl2, shifted the pattern toward the K-type. Na-type digestion caused a greater loss of K-phosphatase than (Na + K)-ATPase activity, and the residual K phosphatase activity was more sensitive to inhibition by Triton and ATP but stimulated more by dimethyl sulfoxide and inhibited less by Pi and MnCl2; all these effects are consistent with such digestion shifting equilibria toward E1 enzyme states. Accordingly, the K0.5 for K+ to activate the (Na + K)-ATPase was increased. However, the K0.5 for the K-phosphatase was unchanged; this observation requires revision of previous formulations, and bears on additional aspects of enzyme activity as well. PMID- 6298195 TI - The distance between thiol groups in the gamma subunit of coupling factor 1 influences the proton permeability of thylakoid membranes. AB - Spinach chloroplast thylakoids treated in the light with bifunctional maleimides were previously shown to be uncoupled. The increase in proton permeability by these reagents is caused by the cross-linking of an accessible group on the gamma subunit of coupling factor 1 (CF1) to a group that becomes exposed to reaction with maleimides only when the thylakoids are energized. In this study, several bifunctional maleimides, including o-, m-, and p-phenylenebismaleimides, 2,3- and 1,5-naphthalenebismaleimides, and azophenylbismaleimide, were tested for their ability to form cross-links and to uncouple photophosphorylation. These reagents form cross-links from about 6 to 19 A. Each reagent was found to form cross-links in the light and to inhibit photophosphorylation. However, the effectiveness of these compounds as uncouplers decreased as the distance between the cross-linked groups increased, indicating that the distance between two groups on the gamma subunit of CF1 can regulate proton flux through the membrane. Monofunctional maleimides cause a light-dependent energy transfer type of inhibition of photophosphorylation. Although this inhibition was correlated to the reaction of the maleimide with a group on the gamma subunit that is exposed only in energized thylakoids, the accessible group on this subunit was also modified by the reagent. However, we show here that the accessible group plays no role in the inhibition of photophosphorylation. This group may be blocked by incubating thylakoids in the dark with methyl methanethiolsulfonate. The light-dependent inhibition of photophosphorylation by N-ethylmaleimide was unaffected by this treatment or by the subsequent removal of the methanethiol moiety from the accessible group. PMID- 6298196 TI - Isolation of totally inverted submitochondrial particles by sonication of beef heart mitochondria. AB - A novel procedure for isolating totally inverted preparations of submitochondrial particles by sonication of beef heart mitochondria is described. The procedure involves only differential centrifugation in 0.25 M sucrose containing 0.15 M KCl. The submitochondrial particles have 96% of their cytoplasmic face cytochrome c-binding sites sequestered within the particles. Mild sonication exposes cytochrome c-binding sites to the medium. The oligomycin-sensitive ATPase of sonic-derived submitochondrial particles, like that of electron transport particles, is inhibited 98% by exogenous isolated ATPase inhibitor protein. NADH oxidase activity in these particles is inhibited by oligomycin. The respiratory control index (uncoupled rate/oligomycin-inhibited rate) is approximately 3.4 and can be increased by washing the particles with medium containing bovine serum albumin. PMID- 6298198 TI - Generation of potential in lipid bilayer membranes as a result of proton-transfer reactions in the unstirred layers. AB - The addition of an uncoupler in the presence of a concentration gradient of weak acids or bases (sodium acetate and ammonium chloride) leads to the generation of a potential on lipid bilayer membranes (LBM) which is positive in sign on the side of the membrane with a high concentration of sodium acetate and negative on the side with a high concentration of ammonium chloride. It is shown that the potential was caused by the pH gradient in the unstirred layers. These effects can be understood in terms of the previously described [Science, 182, 1258 (1973)] model for the transfer of weak acids and bases through LBM. This system described may be useful for quantitation of permeabilities for weak acids and bases through bilayer membranes. PMID- 6298197 TI - Mitochondrial oscillation and activation of H+/cation exchange. AB - Mitochondria incubated aerobically in the presence of tetrapropylammonium and weak acids and in the presence of trace amounts of tetraphenylboron undergo a series of damped oscillations reflecting cycles of osmotic swelling and shrinkage. The matrix volume changes are consequent to transport of tetrapropylammonium catalytically stimulated by tetraphenylboron. The amplitude and frequency of the oscillations increase with the concentration of tetrapropylammonium, as required for critical rates and extents of ion influx. Addition of bovine serum albumin abolishes both the uptake of tetrapropylammonium and the oscillations. Volume oscillations are paralleled by cyclic activation and depression of the respiratory rate. Two lines of evidence suggest that the train of damped oscillations depends on the cyclic activation of an electroneutral exchange of H+ with organic cations rather than on cyclic uncoupling. First, further increase of cation permeability due to a pulse of tetraphenylboron, after initiation of cation efflux, restores cation influx. Second, addition of Mg2+, which abolishes the oscillations, has a much more marked inhibitory effect on the process of cation efflux than on cation influx. Conversely, addition of A23187, which removes membrane-bound Mg2+, promotes cation efflux and thus the oscillations. It is suggested that, in the present system, stretching of the inner membrane and Mg2+ depletion result in activation of an electroneutral H+/organic cation exchange, and that cyclic activation of this reaction results in damped oscillations. PMID- 6298200 TI - In vitro association of a replication complex with a yeast chromosomal replicator. PMID- 6298201 TI - Isolation and characterization of a genomic clone encoding chick alpha 1 type III collagen. AB - We have isolated a cloned segment of the chick alpha 1 type III collagen gene from a genomic DNA library. This DNA clone was obtained by cross-hybridization with a cDNA clone containing sequences coding for a segment of the carboxy propeptide of chick alpha 1 type I collagen. This genomic clone was identified as coding for alpha 1 type III collagen by DNA sequence analysis of two segments of the gene and by its ability to hybridize to crop mRNA but not to calvaria mRNA in agreement with the presence of alpha 1 type III collagen in crop but not in calvaria. The deduced amino acid sequences reveal characteristic features for alpha 1 type III collagen and are in good agreement with the known protein sequences of calf and human alpha 1 type III collagen. The size of alpha 1 type III mRNA is about 6 kilobases, slightly larger than the mRNA for alpha 1 type I. R-loop electron microscopic analysis indicates that at least 16 exons are interspersed in a 9-kilobase DNA segment that contains the third distal part of the alpha 1 type III collagen gene. PMID- 6298199 TI - Localization of endogenous ATPases at the nerve terminal. AB - We have investigated the localization of a set of intrinsic ATPase activities associated with purified synaptic plasma membranes and consisting of (a) a Mg2+ ATPase; (b) an ATPase active at high concentrations of Ca2+ in the absence of Mg2+ (CaH-ATPase); (c) a Ca2+ requiring Mg2+-dependent ATPase (Ca + Mg)-ATPase, stimulated by calmodulin (Ca-CaM-ATPase); (d) a Ca2+-dependent ATPase stimulated by dopamine (DA-ATPase); and (e) the ouabain-sensitive (Na + K)-ATPase. The following results were obtained: (1) All ATPases are largely confined to the presynaptic membrane; (2) the DA-, (Ca + Mg)-, (Ca-CaM)-, and (Na + K)-ATPases are oriented with their ATP hydrolysis sites facing the synaptoplasm; (3) the Mg- and CaH-ATPases are oriented with their ATP hydrolysis sites on the junctional side of the presynaptic membrane and are therefore classified as ecto-ATPases of as yet unknown function. PMID- 6298202 TI - Synthesis, molecular cloning, and restriction analysis of DNA complementary to vitamin D-dependent calcium-binding protein mRNA from rat duodenum. AB - The mRNA coding for rat intestinal calcium-binding protein, a vitamin D3-induced protein (Mr 7500), has been partially purified from growing rat duodenum. Double stranded DNA synthesized from the purified mRNA preparation was inserted into the PstI site of pBR322, using the oligo(dG-dC) tailing procedure. Clones containing DNA complementary to vitamin D-dependent calcium-binding protein mRNA were selected by differential colony hybridization with [32P] cDNA synthesized from enriched or low vitamin D-dependent calcium-binding protein mRNA preparations. Plasmid DNAs from the selected clones were each verified by both a solution hybrid-arrest assay and a filter hybrid-selection assay. Four recombinant clones showed identical endonuclease restriction maps and contained inserts ranging from 250 to 380 base pairs. PMID- 6298203 TI - Stimulation of phosphatidylinositol 4,5-bisphosphate hydrolysis in hepatocytes by vasopressin. AB - Hepatocyte phosphatidylinositol 4,5-bisphosphate (4,5-P2), phosphatidylinositol 4 phosphate (4-P), and phosphatidylinositol were labeled with 3H when rats were injected intraperitoneally with 200 microCi of [2-3H] myo-inositol 18 h previously. Phosphatidylinositol 4,5-P2 and phosphatidylinositol 4-P accounted for 0.84 +/- 0.06 and 7.48 +/- 0.36%, respectively, of the total [3H] myo inositol containing phospholipids. The breakdown of phosphatidylinositol 4,5-P2 was stimulated transiently (maximum effect seen at 15 s) and in a Ca2+-dependent manner by 10(-8) M vasopressin. Phosphatidylinositol 4-P breakdown was enhanced to a smaller, but longer, extent by vasopressin, whereas no changes in phosphatidylinositol were detected up to 120 s. Subcellular fractionation studies also showed no preferential breakdown of phosphatidylinositol in plasma membranes at 5-20 min. Only doses of vasopressin (10(-8) and 10(-7) M) in excess of those producing maximum effects on phosphorylase activation and Ca2+ efflux (10(-9) M) were effective at stimulating phosphatidylinositol 4,5-P2 breakdown. It is concluded that phosphatidylinositol 4,5-P2 breakdown induced by vasopressin in rat hepatocytes is not responsible for the mobilization of Ca2+ which leads to the activation of phosphorylase. On the contrary, it is Ca2+-dependent and appears to require the occupation of more receptors than are required for Ca2+ mobilization and phosphorylase activation. PMID- 6298204 TI - The in vitro expression of the gene for Escherichia coli ADP glucose pyrophosphorylase is stimulated by cyclic AMP and cyclic AMP receptor protein. PMID- 6298205 TI - Modulation of cAMP-mediated differentiation in ovarian granulosa cells by epidermal growth factor and platelet-derived growth factor. PMID- 6298206 TI - Region-specific initiation of mouse mammary tumor virus RNA synthesis by endogenous RNA polymerase II in preparations of cell nuclei. AB - Adenosine 5'-O-(2-thiotriphosphate) (ATP beta S) and guanosine 5'-O-(2 thiotriphosphate) (GTP beta S) were used to demonstrate initiation of mouse mammary tumor virus (MMTV) RNA in preparations of whole nuclei from control and glucocorticoid-treated MMTV-infected rat hepatoma tissue culture cells. RNA chains initiated in the cell-free reaction retain a thiol group at the 5' end and can be separated from thiol-free RNA chains by chromatography on mercury Sepharose. The abundance of MMTV sequences was determined by nucleic acid hybridization with filter-bound DNA representing four different regions of the MMTV genome. About six times more MMTV RNA is initiated with GTP beta S than with ATP beta S. Most of the cell-free initiation of MMTV RNA occurs within or very near a 380-nucleotide section of the proviral long terminal repeat that is the presumptive site of transcription initiation in vivo. The sensitivity of MMTV RNA initiation and synthesis to alpha-amanitin and actinomycin D are characteristic of DNA-directed transcription by RNA polymerase II. Nuclei from glucocorticoid treated cells initiate approximately 10 times more MMTV RNA than nuclei from control cells. PMID- 6298208 TI - LDHk, the lactate dehydrogenase associated with transformation by the Kirsten sarcoma virus: a re-evaluation. AB - A lactate dehydrogenase isozyme designated "LDHk" has recently been described in cells transformed by the Kirsten murine sarcoma virus. Several unusual properties were felt to distinguish this LDH isozyme from previously described LDH isozymes. These properties include cathodic electrophoretic migration in acrylamide gels using an imidazole/borate buffer system, inhibition of activity by oxygen and GTP, and stimulation of activity by cyanide. However, this report demonstrates that the muscle-type mammalian LDH isozyme, LDH 5, also exhibits these unusual properties when it is isolated and detected by the same methods previously used to demonstrate LDHk. The unusual properties attributed to LDHk are not intrinsic properties of the enzyme itself but rather result from the methods employed to demonstrate LDHk activity. Numerous similarities exist between LDH 5 and LDHk including electrophoretic mobility, substrate requirements, similar Michaelis constants for lactate, pyruvate, NAD, and NADH, and identical tissue distribution, indicating that LDHk represents mammalian LDH 5. PMID- 6298207 TI - The spectrum of anionic oligosaccharides released by endo-beta-N acetylglucosaminidase H from glycoproteins. Structural studies and interactions with the phosphomannosyl receptor. AB - We have performed a detailed analysis of all the anionic oligosaccharides released by endo-beta-N-acetylglucosaminidase H from the whole cell glycoproteins of P388D1 mouse macrophage-like cells labeled for 14 h with [2-3H]mannose. The major anionic species consisted of phosphorylated high mannose-type oligosaccharides containing one or two phosphomonoesters or phosphodiesters in several different positions. In addition we identified hybrid-type molecules containing one, two, or three sialic acid residues. A subset of the latter molecules also contained phosphodiesters or phosphomonoesters on another branch of the same oligosaccharide. Unlike previously reported hybrid-type molecules, these do not have a "bisecting" N-acetylglucosamine residue on the beta-linked mannose. Some of these oligosaccharides contained an unidentified acid-labile group on the core N-acetylglucosamine or the beta-linked mannose. The glycoproteins secreted by these cells were greatly enriched in hybrid oligosaccharides containing one sialic acid and one phosphomonoester. The interaction of the isolated oligosaccharides with bovine liver phosphomannosyl receptor immobilized on Affigel was analyzed. Oligosaccharides with phosphomonoesters were the only species that interacted with high affinity with the receptor, and molecules with two phosphomonoesters showed the best binding. The location of the phosphomonoester on the oligosaccharide influenced the degree of interaction with the receptor. Removal of accessible nonphosphorylated mannose residues improved the binding in some cases. These findings indicate that the generation of the physiological phosphomannosyl ligand on lysosomal enzymes involves removal of the blocking N-acetylglucosamine residues, trimming of certain mannose residues, and correct positioning of the phosphate esters. PMID- 6298209 TI - Specific desensitization to tumor-promoting phorbol esters in mouse pituitary cells. Evidence that desensitization is a two-step process. PMID- 6298210 TI - Labeling of subunit b of the ATP synthase from Escherichia coli with a photoreactive phospholipid analogue. AB - Purified ATP synthase (F1F0) from Escherichia coli K12 was labeled with the hydrophobic photoreactive label 1-palmitoyl 2-(2-azido-4-nitro)benzoyl sn-glycero 3-[3H]phosphocholine in reconstituted proteoliposomes. The F0-subunit b was predominantly labeled. A very low amount of label was detected on the other F0 subunits a and c. The label in subunit b could be traced back by proteolytic digestion to the NH2-terminal fragment 1 to 53 which contains the stretch of hydrophobic amino acid residues 1 to 32. By sequencing the intact protein, the distribution of label among the amino acids in this segment was determined. Cysteine 21 was predominantly labeled. Other labeled amino acids occurred at the NH2-terminal (Asn-2) and at position 26 (tryptophan). Due to the restricted mobility of the label in the lipid bilayer, these residues are suggested to be located in or close to the polar head of the lipid bilayer. These results will be compared with predictions for the arrangement of the polypeptide b derived from the hydrophobicity profile. PMID- 6298211 TI - Evidence for a repeating 3,4-dihydroxyphenylalanine- and hydroxyproline containing decapeptide in the adhesive protein of the mussel, Mytilus edulis L. AB - Previous work has shown that the permanent adhesive of the marine mussel Mytilus edulis is a protein containing large amounts of hydroxyproline (13%) and 3,4 dihydroxyphenylalanine (Dopa, 11%). The protein also known as the polyphenolic protein is produced and stored in the exocrine phenol gland of the mussel and deposited onto marine surfaces by the animal's foot during the formation of new adhesive plaques. The adhesive protein has been purified by a combination of ion exchange on sulfonylpropyl-Sephadex and gel filtration on low surface energy chromatographic media. Polyacrylamide gel electrophoresis of the protein at acidic pH shows it to consist of two components having a molecular weight of about 130,000. Treatment of the protein with clostridial collagenase reduced the molecular weight by less than 10%. The collagenase-resistant fragment contains most or all of the Hyp and Dopa. Trypsin treatment of the polyphenolic protein results in extensive degradation. The major tryptic peptide (80%) contains 10 amino acids including Hyp and Dopa and was shown by sequence analysis to be H2N Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Dopa-Lys-COOH. Calculations suggest that this and related sequences may be repeated as often as 75 times in the polyphenolic protein. PMID- 6298212 TI - Self-aggregation, a new property of cardiac fatty acid-binding protein. Predictable influence on energy production in the heart. AB - The binding isotherm of 2-(10-carboxydecyl)-2-hexyl-4,4-dimethyl-3 oxyloxazolidine to the 12,000 molecular weight cytoplasmic fatty acid-binding protein purified from pig's heart and as analyzed by electron spin resonance is shown to be sigmoidal and dependent on protein concentration. Its maximum binding capacity, Vmax, is 9.81 microM X g-1 when the protein concentration is 1.98 g . liter-1 and 14.33 microM . g-1 when the concentration is 0.198 g . liter-1. The molar ellipticity, theta, measured by circular dichroism at 225 nm in the protein range of 0.2-3 g . liter-1 is also shown to depend on concentration, with two maxima at 1.7 and 1.2 g . liter-1. The rotational correlation time, tau eff, of a spin-labeled N-(2,2,5,5-tetramethyl-3-carbonyl-pyrroline-1-oxyl)imidazole specifically bound to tyrosine residues of the protein surface, as analyzed by electron spin resonance, is decreasing as a function of protein concentration. To account for these three concentration-dependent variations, we suggest that the protein self-aggregates since these variations will not be found if the protein is monomeric. A very significant influence of this aggregation property of the protein on the activity of fatty acid-dependent membrane-bound enzymes is predicted by a model analysis and, thus, should be considered as a new parameter in the control of energy production in the heart. PMID- 6298214 TI - The zinc-containing high Km cyclic nucleotide phosphodiesterase of bakers' yeast. AB - The high Km cyclic nucleotide phosphodiesterase of Saccharomyces cerevisiae was purified by an improved procedure. Its amino acid composition is reported. Its pI is 5.85 +/- 0.1. Sedimentation equilibrium analysis of the native enzyme gave Mr = 88,000 +/- 6,000, whilst gel electrophoresis in the presence of dodecyl sulfate gave a molecular weight of 43,000, indicating that the enzyme is a dimer. Preparations of 94 +/- 4% purity contained about 2.4 atoms of zinc/43,000 daltons. Inactivation of the enzyme by 8-hydroxyquinoline was accompanied by removal of about 2 zinc atoms per monomer. Partially inactivated enzyme regained activity during dialysis against zinc, or, with less effect, cobalt salts. 8 Hydroxyquinoline (Ki = 1.1 mM) and 1,10-phenanthroline (Ki = 0.6 mM) were competitive inhibitors. The enzyme was also inhibited by the nonchelating 1,7-and 4,7-phenanthrolines and by thiols and KCN, but not by NaN3. These inhibitors probably act by binding to, but not chelating, enzyme-bound zinc. PMID- 6298213 TI - Interaction of epidermal growth factor-dependent protein kinase with endogenous membrane proteins and soluble peptide substrate. PMID- 6298215 TI - Induction of 11 beta-hydroxylase by corticotropin in primary cultures of bovine adrenocortical cells. PMID- 6298216 TI - Binding and degradation of nerve growth factor by PC12 pheochromocytoma cells. AB - The specific binding of various concentrations of 125I-labeled nerve growth factor (NGF) to PC12 cells at 37 degrees C reached maxima after 90 min and then declined to 25% of maximal binding after 10 h. Decreased binding was accompanied by degradation of 125I-NGF and the appearance of acid-soluble biologically inactive 125I (mainly 125I-monoiodotyrosine) in the medium as well as a decrease in the number of surface NGF receptors. The time-dependent decrease in binding and the degradation of 125I-NGF were inhibited by low temperature and the lysosomotropic agent chloroquine while degradation was inhibited by metabolic energy inhibitors in the absence of glucose. Chloroquine also produced an increase in the accumulation of 125I-NGF which was not readily removed from the cells. These data suggest that 125I-NGF bound to PC12 cells is efficiently internalized by receptor-mediated endocytosis and degraded by the lysosomes. It appears from other data that this process does not produce the intracellular signals regulating neurite outgrowth. PMID- 6298217 TI - Electron paramagnetic resonance study of the interaction of some anionic ligands with oxidized Pseudomonas cytochrome oxidase. PMID- 6298218 TI - Nucleotide sequence of the metL gene of Escherichia coli. Its product, the bifunctional aspartokinase ii-homoserine dehydrogenase II, and the bifunctional product of the thrA gene, aspartokinase I-homoserine dehydrogenase I, derive from a common ancestor. PMID- 6298219 TI - Desensitization of the beta-adrenergic receptor of frog erythrocytes. Recovery and characterization of the down-regulated receptors in sequestered vesicles. PMID- 6298221 TI - Adenosine diphosphoribose transfer reactions catalyzed by Bungarus fasciatus venom NAD glycohydrolase. AB - Reactions catalyzed by purified Bungarus fasciatus venom NAD glycohydrolase were demonstrated to include ADP-ribose transfer from NAD to alcohols and to imidazole derivatives to produce a variety of ADP-ribosides. The formation of products was monitored by high performance liquid chromatography. In the enzyme-catalyzed alcoholysis of NAD, the ratio of n-alkyl-ADP-riboside formed to the hydrolytic product, ADP-ribose, increased linearly with alcohol concentration. The effectiveness of alcohols as acceptors of the ADP-ribose moiety in these reactions increased with increasing chainlength of the alcohol used. Linear positive chainlength effects extended from methanol to pentanol suggesting facilitation of these reactions by nonpolar interactions. In the methanolysis reaction, NADP, thionicotinamide adenine dinucleotide, nicotinamide-1, N6 ethenoadenine dinucleotide, and 3-acetylpyridine adenine dinucleotide were shown to be as effective as NAD as donor substrates. The NAD glycohydrolase-catalyzed ADP-ribose transfer to pyridine bases to form NAD analogs was studied at pyridine base concentrations above those determined to be saturating for the base exchange reaction. Under these conditions, the ratio of base exchange to hydrolysis of NAD was directly related to the pKa of the ring nitrogen of the pyridine base employed. In addition to alcoholysis and pyridine-base exchange reactions, the snake venom enzyme was demonstrated to catalyze an ADP-ribose transfer reaction to imidazole derivatives. Arginine methyl ester was ineffective as an ADP-ribose acceptor molecule in these reactions. PMID- 6298220 TI - Purification and characterization of a murine basement membrane collagen degrading enzyme secreted by metastatic tumor cells. AB - A type IV collagen-degrading enzyme activity secreted by a highly metastatic mouse tumor was purified by concanavalin A- and type IV collagen-agarose affinity chromatographies followed by gel filtration on Bio-Gel A-0.5 m. The apparent molecular weight of the enzyme was 160,000 but about 70,000 when Triton X-100 was added to the column buffer. The purified enzyme protein was resolved on sodium dodecyl sulfate-polyacrylamide gel electrophoresis into two polypeptide chains of about 68,000 and 62,000 daltons. The enzyme activity could be increased by preincubation with trypsin and it is possible that the two chains represent latent and active enzyme forms. The enzyme activity was not reduced in the presence of dithiothreitol, it had a pH optimum of 7.6 and was inhibited by EDTA but not N-ethylmaleimide, phenylmethylsulfonyl fluoride, or Trasylol. The inhibition with EDTA was reversible. The pro-alpha 1(IV) and pro-alpha 2(IV) chains of the type IV procollagen substrate were both degraded at a similar rate to form two pairs of degradation fragments corresponding in molecular weights to about 70 and 30% of the original size chains. The presence of Triton X-100 increased slightly the activity of the enzyme and diminished the reduction of its activity upon freezing, indicating that the enzyme is a hydrophobic protein. PMID- 6298222 TI - The amino acid sequence of the beta-subunit of ATP synthase from bovine heart mitochondria. AB - The amino acid sequence of the beta-subunit of bovine heart mitochondrial ATP synthase has been determined by protein sequence analysis. The polypeptide chain of 478 amino acids is blocked at its NH2 terminal. Comparison of this sequence with sequences of the corresponding proteins from Escherichia coli (Saraste, M., Gay, N.J., Eberle, A., Runswick, M.J., and Walker, J.E. (1981) Nucleic Acids Res. 9, 5287-5296) and maize and spinach chloroplasts Krebbers, E.T., Larrinua, I. M., McIntosh, L., and Bogorad, L. (1982) Nucleic Acids Res. 10, 4985-5002; Kurawski, G., Bottomley, W., and Whitfield, P.R. (1982) Proc. Natl. Acad. Sci. U.S.A. 79, 6260-6264) shows that the protein is highly conserved. 70% of residues are identical in E. coli and beef mitochondria. This contrasts with some of the other subunits in the enzyme complex which are much less conserved. PMID- 6298223 TI - Effects of Cu2+-o-phenanthroline on gastric (H+ + K+)-ATPase. Evidence for opening of a closed anion conductance by S-S cross-linkings. AB - Effects of the S-S cross-linking reagent, Cu2+-o-phenanthroline (CuP), on salt conductances of gastric vesicle membranes in which the (H+ + K+)-ATPase is present were studied. CuP caused a dose-dependent increase in the KCl conductance of the vesicle membrane. The increase of the KCl conductance caused by 10 microM CuP was completely prevented by 0.3 mM ATP or 0.3 mM adenyl 5'-yl imidodiphosphate and partially prevented by ADP. The NaCl conductance was also increased by the CuP reaction. However, CuP has no effect on the K2SO4 conductance. Pretreatments of vesicles with 0.1 mM 4-acetoamide-4' isothiocyanostilbene-2,2'-disulfonate, an anion channel inhibitor, completely blocked the effect of CuP. Thus, these effects of CuP are ascribable to the increase in the anion conductance of the vesicle membrane produced by S-S cross linking. Furthermore, tyrosine-tyrosine cross-linking with tetranitromethane also increases the anion conductance. Probable roles of the opening of the closed anion channel of the ATPase were discussed in regard to the acid secretory mechanism of gastric mucosa. PMID- 6298224 TI - Na+ and K+ transport damage induced by oxygen free radicals in human red cell membranes. PMID- 6298225 TI - Isolation of the luteinizing hormone-chorionic gonadotropin receptor in high yield from bovine corpora lutea. Molecular assembly and oligomeric nature. PMID- 6298226 TI - Physical characterization of two-iron uteroferrin. Evidence for a spin-coupled binuclear iron cluster. PMID- 6298227 TI - Intracellular dissociation of receptor-bound asialoglycoproteins in cultured hepatocytes. A pH-mediated nonlysosomal event. AB - The binding, internalization, and degradation of 125I-asialo-orosomucoid were studied in primary monolayer cultures of rat hepatocytes. Ligand entered the cell bound to the asialoglycoprotein receptor and subsequently dissociated from the receptor intracellularly. Rate coefficients for each of the transitions that constitute the endocytic pathway were computed. Subcellular fractionation on Percoll gradients revealed that prior to localization in lysosomes, 125I-asialo orosomucoid resided in a fraction of slightly lower buoyant density than plasma membranes. Neither ammonium chloride (20 mM) nor leupeptin (0.1 mg/ml) affected ligand binding or internalization of prebound ligand. However, both reagents inhibited degradation of ligand by greater than 95%. Of the two, only ammonium chloride inhibited receptor-ligand dissociation. Ammonium chloride treatment resulted in the accumulation of ligand in the prelysosomal fraction. In contrast, exposure of cells to leupeptin led to accumulation of ligand within lysosomes. The results are interpreted in terms of pH-mediated dissociation of ligand receptor complex within a nonlysosomal endocytic vesicle. PMID- 6298229 TI - RNA polymerase I-dependent selective transcription of yeast ribosomal DNA. Identification of a new cellular ribosomal RNA precursor. AB - Selective transcription of hybrid plasmids containing yeast rDNA was achieved with a template-dependent S100 fraction from whole cell extracts of Saccharomyces cerevisiae. A small region of the yeast rDNA which directs selective initiation in vitro was identified by subcloning. An initiation site was mapped within this region on the basis of the molecular weights of transcripts synthesized in vitro from templates which were cleaved with restriction endonucleases at a series of sites downstream from the site of initiation. The initiation site maps to a position 3.0 kilobase pairs upstream from the sequences which encode the 5' terminus of 18 S rRNA. In vitro initiation from this site is not inhibited by 50 micrograms/ml of alpha-amanitin and is completely abolished when the reactions contain 0.2 M (NH4)2SO4. Based on these data, selective transcription of yeast rDNA in vitro is RNA polymerase I-dependent. Several S1 nuclease-resistant hybrids are formed between DNA probes labeled at restriction endonuclease sites downstream from the in vitro initiation site and high molecular weight cellular RNA. The 5' terminus of the most abundant rRNA precursor maps approximately 0.7 kilobase pair upstream from sequences which encode the 5' terminus of 18 S rRNA. This corresponds to the 5' terminus of the 35 S rRNA precursor reported by others. The 5' terminus of the largest detectable precursor synthesized in vivo corresponds closely with the initiation site identified in vitro. Based on the data presented here, RNA polymerase I traverses the interspersed 5 S rRNA gene. Since these two ribosomal genes are transcribed in opposite directions, this arrangement of the RNA polymerase I and III promoters may ensure that equivalent amounts of the two gene products are synthesized in vivo. PMID- 6298228 TI - Purification of yeast RNA polymerases using heparin agarose affinity chromatography. Transcriptional properties of the purified enzymes on defined templates. AB - A rapid procedure for the simultaneous purification of yeast RNA polymerases I, II, and III is described. The procedure involves direct fractionation of a yeast cell extract by heparin agarose affinity chromatography, followed by glycerol gradient centrifugation and DEAE-Sephadex chromatography. The purification can be completed in 3-4 days using 20-200 g of yeast cells. Two forms each of RNA polymerases I, II, and III are resolved after DEAE-Sephadex chromatography. In the cases of RNA polymerases I and II, these forms differ in subunit structure. The transcriptional properties of the isolated enzymes were determined using hybrid plasmid DNA templates containing yeast ribosomal and glycolytic structural genes. Both forms of RNA polymerases I and II transcribe plasmid DNA templates with low efficiency and no evidence for selective initiation of transcription was found for these enzymes using a wide variety of templates. Both forms of RNA polymerase III transcribe plasmid DNA templates with high efficiency and direct the synthesis of discrete transcripts. Sites for initiation and termination of transcription by RNA polymerase III within defined plasmid DNA templates were determined. The data show that RNA polymerase III-dependent synthesis of discrete transcripts from restriction endonuclease-digested plasmid DNA templates is initiated from selected ends of the templates and terminates at discrete sites downstream from the site of initiation. RNA polymerase III initiates synthesis at many sites within supercoiled plasmid DNA templates. PMID- 6298230 TI - Purification and characterization of a type II casein kinase from Drosophila melanogaster. AB - A cyclic nucleotide-independent protein kinase has been isolated from Drosophila melanogaster by chromatography on phosphocellulose and hydroxylapatite followed by gel filtration and glycerol gradient sedimentation. As determined by sodium dodecyl sulfate gel electrophoresis, the purified enzyme is greater than 95% homogeneous and is composed of two distinct subunits, alpha and beta, having Mr = 36,700 and 28,200, respectively. The native form of the enzyme is an alpha 2 beta 2 tetramer having a Stokes radius of 48 A, a sedimentation coefficient of 6.4 S, and Mr approximately 130,000. The purified kinase undergoes an autocatalytic reaction resulting in the specific phosphorylation of the beta subunit, exhibits a low apparent Km for both ATP and GTP as nucleoside triphosphate donor (17 and 66 microM, respectively), phosphorylates both casein and phosvitin but neither histones nor protamine, modifies both serine and threonine residues in casein, and is strongly inhibited by heparin (I50 = 21 ng/ml). These properties are remarkably similar to those of casein kinase II, an enzyme previously described in several mammalian and avian species. The strong similarities among the insect, avian, and mammalian enzymes suggest that casein kinase II has been highly conserved during evolution. PMID- 6298231 TI - Loss of the inhibitory function of the guanine nucleotide regulatory component of adenylate cyclase due to its ADP ribosylation by islet-activating protein, pertussis toxin, in adipocyte membranes. AB - Adenylate cyclase of rat adipocyte membranes exhibited dual responses in a strictly GTP-dependent manner; an activation took place in the presence of certain receptor agonists such as isoproterenol or secretin, whereas an inhibitory phase was observed with other agonists such as prostaglandin E1 or purine-modified adenosine as well as with the stimulatory agonists at higher GTP concentrations. Treatment of membrane donor cells with islet-activating protein (IAP), pertussis toxin, abolished the inhibitory phase while preserving the activatory phase. This unique action of IAP was associated with ADP-ribosylation of a membrane Mr = 41,000 protein. In contrast, the inhibitory phase was preserved in membranes from cholera toxin-treated cells. Monophasic and persistent activation of the cyclase was provoked by guanyl-5'-yl beta,gamma imidodiphosphate. The time lag normally observed for the guanyl-5'-yl beta,gamma imidodiphosphate activation was decreased by isoproterenol or cholera toxin but was not altered by IAP treatment. Our conclusion is that the sole site of IAP action is the guanine nucleotide regulatory protein (Ni) that is required for transmission of inhibitory signals from receptors to the catalytic unit of adenylate cyclase; the function of Ni is lost upon IAP-catalyzed ADP ribosylation of the Mr = 41,000 protein which appears to be an active subunit of Ni. A possibility is discussed that rather diverse effects of IAP so far reported with various cell types are accounted for in terms of such interference with the function of Ni. PMID- 6298232 TI - Limited proteolysis studies on the Escherichia coli single-stranded DNA binding protein. Evidence for a functionally homologous domain in both the Escherichia coli and T4 DNA binding proteins. AB - Limited proteolysis can be used to remove either 42 or 62 amino acids at the COOH terminus of the 18,873-dalton Escherichia coli single-stranded DNA binding protein (SSB). Since poly(dT), but not d(pT)16, increases the rate of this reaction, it appears that cooperative SSB binding to single-stranded DNA (ssDNA) is associated with a conformational change that increases the exposure of the COOH terminus to proteolysis. As a result of this DNA-induced conformational change, we presume that the COOH-terminal region of SSB will become more accessible for interacting with other proteins that utilize the SSB:ssDNA complex as a substrate and that are involved in E. coli DNA replication, repair, and recombination. Removal of this COOH-terminal domain from SSB results in a stronger helix-destabilizing protein which suggests this region may be important for controlling the ability of SSB to denature double-stranded DNA. Since similar results have previously been reported for the bacteriophage T4 gene 32 protein (Williams, K.R., and Konigsberg, W. (1978) J. Biol. Chem. 253, 2463-2470; Hosoda, J., and Moise, H. (1978) J. Biol. Chem. 253, 7547-7555), the acidic, COOH terminal domains of these two single-stranded DNA binding proteins may be functionally homologous. Preliminary evidence is cited that suggests other prokaryotic and eukaryotic DNA binding proteins may contain similar functional domains essential for controlling their ability to invade double helical DNA. PMID- 6298233 TI - The nucleotide sequence of rat liver fatty acid binding protein mRNA. AB - Rat liver fatty acid binding protein mRNA is present in both liver and intestinal epithelium. It is the most abundant mRNA in the small intestinal mucosa. We have determined the sequence of this mRNA from cloned cDNA. It consists of a 5' nontranslated domain at least 39 nucleotides long, a coding region which specifies a 127-amino acid, 14,184-dalton polypeptide, and a 3'-nontranslated region 68 nucleotides long. An AAUAAA sequence begins 21 nucleotides before the 3'-terminal poly(A) tail. The derived protein sequence was verified by automated sequential Edman degradation of (i) the primary translation product of the mRNA and (ii) a cyanogen bromide peptide generated from the mature liver protein. Evidence is presented which suggests that this protein is targeted to the cytoplasm. The NH2 terminus of the primary translation product is acetylated in vitro. No cleavable signal peptide sequence or internal signal sequence equivalent was detectable in a co-translational cleavage assay. Cloned cDNA was used to establish that this mRNA was 12 times more abundant in intestinal epithelial than in liver RNA. PMID- 6298234 TI - Purification of the receptor for nerve growth factor from A875 melanoma cells by affinity chromatography. AB - The receptor for nerve growth factor (NGF) has been purified to near homogeneity from octylglucoside extracts of A875 melanoma cell membranes by the use of repetitive affinity chromatography on NGF-Sepharose. Elution of purified receptor (NGF receptor) was accomplished with 0.15 M NaCl, pH 11.0, containing phosphatidylcholine and octylglucoside. Chromatography on two columns of NGF Sepharose yielded a 1500-fold purification of the receptor, as assessed by 125I NGF binding, and permitted recovery of 9% of the total binding activity in the soluble extract. Scatchard analysis of equilibrium binding of 125I-NGF provided similar Kd values for NGF receptors in soluble extracts of A875 membranes (2.2 nM) and with purified NGF receptor (3.1 nM). Examination of NGF receptor after electrophoresis on sodium dodecyl sulfate-polyacrylamide gels revealed the presence of two major peptides, of Mr = 85,000 and Mr = 200,000. Affinity labeling experiments, done with 125I-NGF and A875 cells, soluble extracts of A875 cell membranes, and purified receptor, show that both of these components of the NGF receptor can be specifically cross-linked to 125I-NGF. PMID- 6298235 TI - Diversity of beta-crystallin mRNAs of the chicken lens. Hybridization analysis with cDNA clones. PMID- 6298236 TI - Effects of butyrate on histone deacetylation and aldosterone-dependent Na+ transport in the toad bladder. PMID- 6298237 TI - Fibronectin receptors from Staphylococcus aureus. PMID- 6298238 TI - The effects of 6-hydroxydopamine or (-)-noradrenaline treatment in vitro on noradrenergic transmission in the rat anococcygeus muscle. PMID- 6298239 TI - Desensitization of beta-adrenoreceptors in guinea-pig trachea: a prostaglandin mediated phenomenon. AB - 1 The formation and release of PGE2-like material (PGE2-lm) from guinea-pig isolated trachea and human bronchi following relaxation with isoprenaline (I) was investigated. 2 When airway smooth muscle precontracted with pilocarpine is relaxed by I, PGE2-lm is released in the bathing fluid. Under conditions of beta adrenoreceptor desensitization, a greater amount of PGE2-lm is formed and the responsiveness of guinea-pig tracheal spirals to cumulative doses of I is diminished. 3 Inhibition of cyclo-oxygenase with indomethacin does not modify the relaxation induced by I but prevents the occurrence of refractoriness to I as well as formation and release of PGE2-lm. Addition of exogenous PGE2 to guinea pig tracheas which were treated with indomethacin is able to restore the capacity of I to cause tachyphylaxis. 4 It is concluded that PGE2 which does not mediate the relaxation induced by I in airway smooth muscle, is responsible for the onset of desensitization of the beta-adrenoreceptor. PMID- 6298240 TI - Modulation by beta-adrenoreceptors of spontaneous contractions of rat urinary bladder. AB - 1 Propranolol (0.1 mg/kg i.v.) but not metoprolol (0.2 mg/kg i.v.) pretreatment increased the spontaneous motility triggered by progressive filling of rat urinary bladder without a concomitant effect on bladder capacity, except at high filling volumes. Compared to controls, the spontaneous motility of urinary bladder in propranolol pretreated rats displayed a higher frequency, indicating the existence of a tonic sympathetic inhibition. 2 beta-adrenoreceptor stimulation by isoprenaline (0.1-10 microgram/kg i.v.) or terbutaline (0.1-1 mg/kg i.v.) in vivo produced a dose dependent inhibition of bladder spontaneous motility which was antagonized by propranolol (0.1 mg/kg i.v.) but not by metoprolol (0.2 mg/kg i.v.). Propranolol (0.2 mg/kg i.v.) pretreatment did not antagonize the inhibition of bladder motility produced by intravenous papaverine (0.5 mg/kg). 3 Propranolol (0.1 mg/kg i.v.) significantly antagonized the isoprenaline-induced tachycardia (beta 1 mediated) and fall in diastolic blood pressure (beta 2 mediated) while metoprolol (0.2 mg/kg i.v.) antagonism was confined to beta 1 mediated responses. 4 Isoprenaline (0.25-1.5 microM) inhibited in a concentration dependent manner field stimulation induced contractions of rat detrusor muscle strips as did tetrodotoxin (0.5 microM). Hexamethonium (50 microM) had no inhibitory effect. 5 Our in vivo findings support the view that beta 2-adrenoreceptors are responsible for modulating bladder motility mainly by suppressing the onset of spontaneous contractions. PMID- 6298241 TI - Effects of prostaglandin(PG) E2, D2, I2, 6-keto-PGF1 alpha and arachidonic acid on excitatory transmission of canine small intestine. PMID- 6298242 TI - Preliminary report on the osteogenic potential of a biodegradable copolymer of polyactide (PLA) and polyglycolide (PGA). AB - A biodegradable copolymer of 50 polylactide: 50 polyglycolide was prepared for implantation into experimentally created osseous defects in the tibias of 25 rats. Similarly prepared defects were made in the humeri of the same rats and these defects did not receive copolymer implants. Upon sacrifice, both the implant treated and untreated sites of the experimentally produced osseous defects were evaluated by gross appearance and by histomorphometric examination using a Zeiss Videoplan Image Analysis System with Osteoplan (version 4.1). The animals were evaluated in groups of five at 7, 14, 21, 28, and 42 days. When compared with bony defects that were not treated with the biocompatible, biodegradable copolymer implant, the implant sites displayed an accelerated rate of healing at 7, 14, 21, and 28 days (p less than 0.001). A similar healing response rate, however, was observed at 42 days (p less than 0.25-0.1). No adverse host tissue responses were observed histologically. PMID- 6298243 TI - Circadian variability of kinetics in a transplantable islet-cell tumor of the golden hamster. AB - The kinetics of a transplantable islet-cell tumor known to secrete paranormal hormonal compounds without any influence of usual regulatory mechanisms was studied in the golden hamster. It was found that during the 42nd diurnal period of growth, labelling and mitotic indices (peaks at midday), mean grain counts (lower at 4 a.m. and 8 a.m.), and PLM data (shift between G1 and S phase durations) varied significantly. The results are discussed in view of the discrepancy between hormonal release (unaffected by normal triggering factors) and circadian kinetics indicating the persistence of tumoral sensitivity to some control mechanisms. The theoretical importance of the diurnal variability for the correct interpretation of the internal kinetics of tumors is recalled. PMID- 6298244 TI - Synthesis and evaluation of catechol analogs of diethylstilbestrol on a hormone dependent human mammary carcinoma implanted in nude mice. AB - 3,4-Bis(3',4'-diacetoxyphenyl)hex-3-ene (5a), 4(4'-acetoxyphenyl)-3(3',4' diacetoxyphenyl)hex-3-ene (5b), and 4(3'-acetoxyphenyl)-3(3',4' diacetoxyphenyl)hex-3-ene (5c) were synthesized according to the method of Dodds et al. (1939). All compounds inhibited the interaction of 3H-estradiol with its receptor. The relative binding affinity (RBA) values increased in the order: 5a (1.0) less than 5c (7.6) less than 5b (21.7). In the immature mouse uterine weight bioassay, the uterotrophic activity of 5a-c was only weak. 5a and 5c, but not 5b, exhibited significant antiuterotrophic properties. All compounds significantly inhibited the growth of a postmenopausal hormone-dependent human mammary carcinoma serially implanted in nude mice. PMID- 6298245 TI - Lung cancer in Greenland--selected epidemiological, pathological, and clinical aspects. AB - One hundred and seven cases of lung cancer were first diagnosed among indigenous Greenlanders during the 25 years from 1955 to 1979. Compared with the Danish population, relative risk of lung cancer among male Greenlanders increased from 0.4 in 1955-1959 to 1.1 in 1975-1979. Relative risk among women increased from 0.7 to 2.1. The age-adjusted rate for women during the period 1975-1979 is one of the highest on record. This increase in lung-cancer risk followed a sharp rise in the use of cigarettes in the general population. An association with cigarette consumption was also suggested by tumor histology and location. A synergistic role of other risk hazards is possible. Local factors arising from previous tuberculosis involvement may have favored lung cancer development in men. More than 2/3 of the cases of lung cancer diagnosed between 1955 and 1974 were brought to medical attention because of clinical symptoms although most of the population was screened annually or biannually for tuberculosis. Intervention through screening had little impact on the overall very low survival rate. PMID- 6298246 TI - The correlation between tissue differentiation and production of mammary tumor virus (MTV) in transplanted murine mammary tumors. Electron microscopic observations. AB - The spontaneous mammary tumors of the NMRI mouse are well developed microcystic adenocarcinomas. Serial isologous transplantation of the tumors results in nearly complete dedifferentiation to a solid tumor, in which only electron microscopically rudimentary acinus-like microlumina can be observed. The adenocarcinomas produce A and B particles in abundance, with the A particles appearing intracellularly in the adluminal cytoplasmic regions of the epithelial cells in association with typical cellular structures and the B particles being restricted to closed extracellular compartments such as vacuoles or acini alone. The loss of alveolar organization in the solid tumors is followed by an almost complete reduction in mature B particles, while A particles are still regularly observed and appear to be less reduced in number. This suggests that the production of extracellular B particles is dependent upon the secretory activity of the tumor cells and that in nonsecreting cells it is predominantly a late step in virus release that is inhibited, not the synthesis of intracellular precursors. PMID- 6298248 TI - Yeast mannans inhibit binding and phagocytosis of zymosan by mouse peritoneal macrophages. AB - We have examined the effects of various mannans, glycoproteins, oligosaccharides, monosaccharides, and sugar phosphates on the binding and phagocytosis of yeast cell walls (zymosan) by mouse peritoneal macrophages. A phosphonomannan (PO(4):mannose ratio = 1:8:6) from kloeckera brevis was the most potent inhibitor tested; it inhibited binding and phagocytosis by 50 percent at concentrations of approximately 3-5 mug/ml and 10 mug/ml, respectively. Removal of the phosphate from this mannan by mild acid and alkaline phosphatase treatment did not appreciably reduce its capacity to inhibit zymosan phagocytosis. The mannan from saccharomyces cerevisiae mutant LB301 inhibits phagocytosis by 50 percent at 0.3 mg/ml, and a neutral exocellular glucomannan from pichia pinus inhibited phagocytosis by 50 percent at 1 mg/ml. Cell wall mannans from wild type S. cervisiae X2180, its mnn2 mutant which contains mannan with predominantly 1(arrow)6- linked mannose residues, yeast exocellular mannans and O phosphonomannans were less efficient inhibitors requiring concentrations of 1-5 mg/ml to achieve 50 percent reduction in phagocytosis. Horseradish peroxidase, which contains high-mannose type oligosaccharides, was also inhibitory. Mannan is a specific inhibitor of zymosan binding and phagocytosis. The binding and ingestion of zymosan but not of IgG- or complement-coated erythrocytes can be obliterated by plating macrophages on substrates coated with poly-L-lysin (PLL) mannan. Zymosan uptake was completely abolished by trypsin treatment of the macrophages and reduced by 50-60 percent in the presence of 10 mM EGTA. Pretreatment of the macrophages with chloroquine inhibited zymosan binding and ingestion. These results support the proposal that the macrophage mannose/N acetylglucosamine receptor (P. Stahl, J.S. Rodman, M.J. Miller, and P.H. Schlesinger, 1978, Proc. Natl. Acad. Sci. U.S.A. 75:1399-1403, mediates the phagocytosis of zymosan particles. PMID- 6298249 TI - Isolation of rat hepatocyte plasma membranes. I. Presence of the three major domains. AB - A rat liver plasma membrane preparation was isolated and characterized both biochemically and morphologically. The isolation procedure was rapid, simple and effective in producing a membrane fraction with the following biochemical characteristics: approximately 40-fold enrichment in three plasma membrane markers, 5'-nucleotidase, alkaline phosphodiesterase I (both putative bile canalicular membrane enzymes), and the asialo-glycoprotein (ASGP) receptor (a membrane glycoprotein present along the sinusoidal front of hepatocytes); a yield of each of these plasma membrane markers that averaged approximately 16%; and minimal contamination by lysosomes, nuclei, and mitochondria, but persistent contamination by elements of the endoplasmic reticulum. Morphological analysis of the preparation revealed that all three major domains of the hepatocyte plasma membrane (sinusoidal, lateral, and bile canalicular) were present in substantial amounts. The identification of sinusoidal membrane was further confirmed when ASGP binding sites were localized predominantly to this membrane in the isolated PM using electron microscope autoradiography. By morphometry, the sinusoidal front membrane accounted for 47% of the total membrane in the preparation, whereas the lateral surface and bile canalicular membrane accounted for 6.8% and 23% respectively. This is the first report of such a large fraction of sinusoidal membrane in a liver plasma membrane preparation. PMID- 6298251 TI - Complementation for the growth response to insulin and MSA occurs at a step distal to hormone-receptor interaction in mouse embryo fibroblast X melanoma cell hybrids. PMID- 6298252 TI - Tunicamycin inhibits the expression of surface Na+ channels in cultured muscle cells. AB - We have investigated the effect of tunicamycin (TM), an inhibitor of protein glycosylation, on surface Na+ channels in cultured chick skeletal muscle cells. The expression of Na+ channels, estimated by the measurement of batrachotoxin (BTX)-activated 22Na+ uptake, was found to be significantly diminished after exposure of muscle cells to TM. This effect is partially reversed by the protease inhibitor leupeptin and is associated with a markedly enhanced rate of disappearance of Na+ channels from the surface of TM-treated cells. Our findings suggest that protein glycosylation contributes to the metabolic stability of voltage-sensitive Na+ channels. PMID- 6298250 TI - Changes in plasma membrane glycoproteins of rat spermatozoa during maturation in the epididymis. AB - Glycoproteins on the plasma membrane of testicular and cauda epididymidal spermatozoa have been labeled with galactose oxidase/NaB [3H]4 and sodium metaperiodate/NaB[3H]4, followed by analysis on SDS polyacrylamide gels. The major glycoprotein labeling on testicular spermatozoa has a molecular weight 110,000 whereas on cauda epididymidal spermatozoa greater than 90% of the radio label is incorporated into proteins of molecular weight 32,000. These 32,000-mol wt X proteins are homologous with proteins of similar molecular weight purified from the epididymal secretion and which have been shown previously to be synthesized in the caput epididymidis under hormonal control. Immunofluorescence revealed that the 32,000-mol wt proteins are present on the flagellum of mature but not immature spermatozoa and that they have a patchy distribution suggesting that they are mobile within the plane of the membrane. The membrane-bound 32,000 mol wt proteins possess hydrophobic domains as revealed by charge-shift electrophoresis and they also label with a lipophilic photoaffinity probe suggesting that they are in contact with the lipid bilayer. The evidence indicates that there is a considerable reorganization of the molecular structure of the plasma membrane of spermatozoa during maturation in the epididymis and that some of the changes are brought about by a direct interaction with epididymal secretory proteins. PMID- 6298253 TI - Biologic activity of anti-thyrotropin anti-idiotypic antibody. AB - We raised an antihuman thyrotropin anti-idiotypic antibody and showed that it was active at the thyrotropin receptor. Thus this antibody inhibited 125I b-TSH binding to thyroid plasma membranes, stimulated adenylate cyclase activity through a guanyl nucleotide-dependent mechanism, increased radioiodide entry rate into isolated porcine thyroid follicular cells, and induced such cultured cells to organize into follicles. All these parameters are typical of thyrotropin action. This work raises the possibility that thyroid stimulating antibodies that cause the hyperthyroidism of Graves disease may be, at least in some patients, anti-thyrotropin anti-idiotypic antibodies. It also offers a novel method whereby antireceptor antibodies used in the isolation and characterization of the receptor may be raised from ligands. PMID- 6298254 TI - Regulation of herpes simplex virus gene transcription in vitro. AB - We used partially purified RNA polymerase II from uninfected (Pol II) and from herpes simplex virus type 1 (HSV-1) infected HEp-2 cells (Pol II-H) to transcribe HSV-1 DNA in vitro. Gel electrophoretic analysis of the products produced from native HSV-1 DNA yielded weight average chain lengths of 4.0 and 3.5 kb for the Pol II and Pol II-H products, respectively. Blot hybridization analyses of the HSV DNA transcripts showed that both enzymes transcribed RNA from essentially all regions of the genome. However, Pol II preferentially transcribed regions coding for the immediate-early or alpha mRNAs, whereas Pol II-H preferentially copied regions coding for the early (beta) and late (gamma) gene products. Transcriptional analyses of the cloned HSV-1 Bam HI-Q fragment (containing the thymidine kinase (TK) gene) and its subfragments showed that (1) the major transcripts produced by Pol II-H were distinctly different from those produced by Pol II; (2) Pol II and Pol II-H utilized different promoters for the synthesis of major transcripts; (3) both enzymes produced three minor transcripts that were partially overlapping and in opposite direction to the TK gene; and (4) only Pol II-H initiated transcription from the TK promoter. In contrast, both Pol II and Pol II-H generated an identical set of transcripts from an adenovirus 2 early region DNA fragment. The sizes of the products suggest that RNA processing may be occurring in vitro. These results show that HSV-1 infection alters the in vitro transcriptional specificity of RNA polymerase II and demonstrate that this system should be useful for studying in vitro the regulation of gene transcription. PMID- 6298255 TI - Analysis of v-mos encoded proteins in cells transformed by several related murine sarcoma viruses. AB - We have used antisera against synthetic peptides to identify and characterize a 37,000 dalton v-mos encoded protein (p37mos) in cells transformed by M-MuSV 124. p37mos, a phosphoprotein, comprises only about 0.0005% of total cellular protein in cell lines transformed by M-MuSV 124. NIH 3T3 cells acutely infected with M MuSV 124, however, contain 30-100-fold more p37mos. These elevated levels of p37mos correlate with striking morphological changes and cell death in the acutely infected cell population. Using the antipeptide antisera, we have extended the analysis of v-mos proteins to include several other MuSV variants that contain a similar v-mos gene to M-MuSV 124. With the exception of P85, the gag-mos fusion protein from ts110 MuSV, the v-mos gene of these variants is expressed as a 35,000-37,000 dalton protein (size depending on the particular virus). PMID- 6298247 TI - Endocytosis and the recycling of plasma membrane. PMID- 6298256 TI - A basement membrane-associated glycoprotein from skeletal muscle. AB - We have isolated a major glycoprotein that appears to be associated with rat skeletal muscle basement membrane. We determined that the glycoprotein was part of the muscle cell surface complex when we found it to be enriched in preparations of muscle ghosts. We isolate the glycoprotein from homogenized muscle preextracted with 4 M and 8 M urea. It elutes as a major component in the presence of 8 M urea/50 mM 2-mercaptoethanol. Its apparent molecular weight on sodium dodecyl sulfate gels is 130,000. Amino acid analysis indicates that it is not a collagen but that it does contain small amounts of hydroxyproline and hydroxylysine. There may be collagenous domains in the glycoprotein molecule, for it is cleaved into three fragments by purified bacterial collagenase. Immunoperoxidase staining confirms that the 130,000-dalton protein is localized at the surface of adult skeletal muscle cells. It is probably a general basement membrane-associated glycoprotein because we found material immunologically cross reactive with the muscle glycoprotein in basement membrane regions of kidney, liver, brain, and small intestine. We have shown the glycoprotein to be distinct from fibronectin, laminin, and types I, III, IV, and V collagens in enzyme-linked immunosorbent assays. PMID- 6298257 TI - Epidermal growth factor stimulates fluid phase endocytosis in human fibroblasts through a signal generated at the cell surface. AB - We have investigated the stimulation of fluid phase endocytosis by epidermal growth factor (EGF) in normal human fibroblasts using 125I-labeled polyvinylpyrrolidone (125I-PVP) as a fluid phase marker. We found that EGF initially induced a threefold increase in the rate of 125I-PVP uptake. This initial burst of fluid uptake terminated within 10 min. Thereafter, the rate of fluid uptake in EGF-treated cells was approximately 40% higher than in control cells. To identify the cellular site of EGF action in stimulating fluid phase endocytosis, we examined the kinetics of the induction of this response as well as the kinetics of cell surface binding and internalization of 125I-EGF. Although there was no detectable lag between binding of EGF to the cell surface and its internalization, the kinetics of the two processes were quite different. Significantly, the kinetics of induction of 125I-PVP uptake matched the kinetics of binding of 125I-EGF to its cell surface receptors, indicating that the signal for the increase in fluid phase endocytosis is generated at the cell surface. To determine if EGF-stimulated fluid phase endocytosis was related to EGF-stimulated endocytosis of its own receptor, we compared the EGF dose dependency and time course of the two processes. Although the stimulated endocytosis of the EGF receptor was not saturable with respect to the concentration of EGF used, the stimulation of fluid phase endocytosis was half maximal at an EGF concentration of 1 ng/ml and saturated at a concentration of 5 ng/ml. Also, the stimulation of fluid phase endocytosis was sevenfold greater initially after adding EGF than after a 30-min continuous incubation with the hormone, whereas the enhanced clearance of the EGF receptor did not change during this time period. We conclude that the EGF-stimulated increase in fluid phase endocytosis is not directly coupled to EGF-stimulated endocytosis of its own receptor but instead to a separate signal generated at the cell surface. PMID- 6298259 TI - Method for determination of angiotensin-converting enzyme activity in blood and tissue by high-performance liquid chromatography. AB - A simplified method for an angiotensin-converting enzyme activity assay in biological samples was developed. Samples were incubated with hippurylhistidylleucine, an artificial substrate of angiotensin-converting enzyme. The reaction was terminated by the addition of metaphosphoric acid and liberated hippuric acid in the supernatant was quantitated directly by reversed phase high-performance liquid chromatography. Tissues were homogenized in the presence of Nonidet-P40, a detergent, and the resulting supernatant was used for the assay of tissue angiotensin-converting enzyme activity by high-performance liquid chromatography. The present procedure made it possible to determine angiotensin-converting enzyme activity in whole blood and the total activity in tissues. A comparative study of angiotensin-converting enzyme activity in plasma, kidney and lung of five experimental animals showed a high degree of variation from species to species. PMID- 6298258 TI - Biochemical characterization of transformation-specific proteins of acute avian leukemia and sarcoma viruses. AB - The biological and biochemical properties of the transformation-specific proteins of three avian oncornaviruses with different oncogenic potentials were compared, namely the gag-myc protein of the avian myelocytomatosis virus MC29, the gag-erb A protein of the avian erythroblastosis virus AEV, and the gag-fps protein of Fujinami sarcoma virus FSV. These oncogenes were analyzed in transformed fibroblasts that expressed only the transforming proteins but showed no virus replication. Monoclonal antibodies against the viral structural protein p19, which is the N-terminus of the proteins, were used for indirect immunofluorescence, for immunoprecipitation of the proteins from subcellular fractions, and for immunoaffinity column chromatography. With this last method a 3000-fold purification of the proteins was obtained. By indirect immunofluorescence it was shown that the gag-myc protein was located in the nucleus, and bound to DNA after purification. The gag-erb A protein was not nuclear but probably located in the cytoplasm and did not bind to DNA after purification. Neither of the two proteins exhibited protein kinase activity. In contrast, the gag-fps protein did not bind to DNA but showed protein kinase activity after purification. It was not located in the nucleus either. PMID- 6298261 TI - Reversed-phase high-performance liquid chromatography of neuropeptides related to adrenocorticotropin, including a potent adrenocorticotropin 4-9 analogue (ORG 2766). PMID- 6298260 TI - Rapid extraction of leukotrienes from biologic fluids and quantitation by high performance liquid chromatography. AB - Previous methods for the recovery and quantitation of leukotrienes have involved tedious extraction procedures, and high-performance liquid chromatographic (HPLC) techniques with significant limitations. We have designed a method to extract leukotrienes from biologic fluids using commercially available silica mini columns requiring minimal preparation. Sample clarification is followed by a sensitive and reproducible HPLC technique which separates and quantifies the leukotrienes LTC4, LTD4, LTB4 (and at least three of their isomers). The entire procedure requires less than one hour per sample. PMID- 6298262 TI - Simultaneous gas chromatographic analysis of drugs of abuse on two fused-silica columns of different polarities. AB - A convenient and rapid gas chromatographic method for analysis of drugs of abuse has been developed. By installing two differently coated columns in a common split-splitless injector and connecting the column ends to a nitrogen-phosphorus detector and a flame ionization detector, considerable and accurate chromatographic information could be obtained from a single run. Quantitative analyses of amphetamine and some other drugs have been performed with the same instrumental set-up. Calibration graphs were evaluated and showed linearity in the concentration range chosen (0.5-10 mg/ml). By using an auto-sampler in combination with a BASIC program, both types of analyses can be performed routinely. PMID- 6298263 TI - Raman spectroscopic detection of haemoproteins in the eluate from high performance liquid chromatography. PMID- 6298264 TI - Evaluation of enzyme-linked immunosorbent assay for quantitation of antibodies to Japanese encephalitis virus in swine sera. AB - Enzyme-linked immunosorbent assay (ELISA) was evaluated for the quantitation of antibodies to Japanese encephalitis virus in swine sera. The assay was highly reproducible (coefficient of variation of the absorbance values obtained with positive sera was less than 3.4%) and was significantly correlated with the conventional haemagglutination inhibition (HI) test (correlation coefficient was 0.944). The statistical analysis based on the frequency distribution of the absorbance values for 366 swine serum samples gave 0.204 as a feasible borderline to differentiate positive sera from negative sera. Using this criterion, all of the sera positive for HI antibody were found positive for antibody by ELISA and also all negative sera by ELISA were negative by HI. Inconsistent results were found in only six cases (1.6%). PMID- 6298265 TI - Rapid methods for extraction and concentration of poliovirus from oyster tissues. AB - A procedure is discussed for the extraction of poliovirus from oyster meats by modification of several enterovirus extraction techniques. The modified method uses meat extract and Cat-Floc, a polycationic electrolyte, for virus extraction and concentration. Virus recovery from inoculated oyster homogenates is 93-120%. Adsorption of viruses to oyster proteins by acidification of homogenates does not affect virus recovery. Elution of viruses from oyster proteins appears more efficient at pH 9.5 than at pH 8.0. This technique is relatively simple, economical and requires only 2.5 h to complete the combined extraction and concentration procedure. PMID- 6298266 TI - Rapid identification and serotyping of poliovirus isolates by an immunoassay. AB - An enzyme-linked immunosorbent assay (ELISA) for the identification and typing of polioviruses is described. Polioviruses could be rapidly detected and typed from cell culture supernatants by a double antibody sandwich technique. The assay is valuable for the rapid screening of a large number of viral isolates from water samples submitted for virological analysis. Several hundred isolates per day can be typed and only the remainder have to be tested by the conventional serum neutralization test. PMID- 6298267 TI - Comparisons of dose-response data for various standard and recombinant DNA derived human interferons. AB - Dose responses of the NIH standard IFN-alpha and IFN-beta preparations were compared with recombinant DNA-derived IFN-beta (IFN-beta 1) and various IFN-alpha subtypes, molecular hybrids and mixtures. Cytopathic effect assays were employed using vesicular stomatitis virus on human HeLa and bovine MDBK cells. A natural peripheral blood lymphocyte and recombinant DNA-derived IFN-gamma were also included in the comparisons. Two-tailed t-tests between slopes showed no significant differences in any pair-wise comparison using crude or highly purified preparations. PMID- 6298268 TI - Primary liver cancer mortality in the United States. AB - We analyzed time trends and regional differences in mortality from primary liver cancer in the United States in order to explore possible effects of several factors which have been suggested as causes for liver cancer. Age-adjusted liver cancer death rates have been stable over time except for nonwhite males among whom there has been a 45% increase in liver cancer mortality between 1958 and 1975. The rates for nonwhite males are twice those for white males, and geographic trends in liver cancer mortality differ by race. The rates for white males are greatest in two southern regions, whereas rates for nonwhite males in these same regions are lower than they are elsewhere. Cirrhosis mortality trends parallel those for liver cancer among nonwhite males but not among white males. The similarities in trends for cirrhosis and liver cancer mortality among nonwhite males suggest that cirrhosis may be a major risk factor in this group. For white males, we must look to some other factor to explain the geographic differences that were observed. PMID- 6298269 TI - Autonomic nervous system abnormalities in cystic fibrosis. AB - The basic defect in cystic fibrosis, the most common lethal genetic diseases of white Americans, is unknown, but the character of the generalized exocrinopathy suggests some disorder of the regulation and control of the process of glandular secretion. Definite abnormalities in all branches of the autonomic nervous system have been demonstrated in patients with cystic fibrosis, including increased sensitivity to alpha-adrenergically stimulated pupillary dilation; increased responsiveness to cholinergic stimulation of pupillary constriction, parotid saliva secretion, and eccrine sweat secretion; and decreased responsiveness to beta-adrenergic stimulation of the cardiovascular system as well as circulating lymphocytes and granulocytes. Since these abnormalities also occur in asymptomatic heterozygotes for cystic fibrosis (parents of patients), they are likely to be inherited characteristics and not secondarily acquired. This constellation of inherited autonomic abnormalities--alpha-adrenergic and cholinergic hyperresponsiveness and beta-adrenergic resistance--may contribute to the pathophysiology of cystic fibrosis and may also be an important clue to the nature of the basic defect. PMID- 6298270 TI - Quantitation by myeloperoxidase assay of neutrophil accumulation at the site of in vivo allergic reactions. AB - Skin window techniques to investigate neutrophil inflammatory reactions in human skin have been limited by cellular distortion and difficulties in quantitation. We have developed a quantitative approach based on the assessment of the myeloperoxidase (MPO) released from sonicated membrane filters to which exuding inflammative cells had adhered. Our studies have shown that (1) a standard curve can be derived for each subject from the linear relationship between amounts of MPO released and designated numbers of blood neutrophils which have been aspirated onto such filters; (2) the number of neutrophils adhering to filters appended to skin window sites can then be reliably estimated by comparing the amount of MPO released by sonication of such filters with the standard curve; and (3) neutrophil accumulation is greater at pollen-challenged than control sites in sensitive subjects. PMID- 6298271 TI - Disk diffusion testing with polymyxin and amikacin for differentiation of Mycobacterium fortuitum and Mycobacterium chelonei. AB - Disk diffusion is one method of susceptibility testing of the Mycobacterium fortuitum complex to antibacterial agents. We utilized disks of polymyxin B (300 U), amikacin, and kanamycin to determine whether they could also be used for species identification when compared with standard biochemical methods. With the polymyxin disk, 100% of 75 M. fortuitum strains produced zones of inhibition, whereas none (0%) of 58 Mycobacterium chelonei subspecies abscessus and chelonei strains had any zone of inhibition. With the amikacin disk, 99% of M. fortuitum biovariant fortuitum had zones of greater than or equal to 30 mm compared with 6% of M. chelonei. The rare M. chelonei-like organisms gave variable results, and 42% of the unnamed "third group" biovariant of M. fortuitum exhibited an unusual but diagnostic pattern of small zone sizes to amikacin and no zone to kanamycin. The kanamycin disk was otherwise not helpful, although it resulted in larger zone sizes for M. chelonei than did amikacin. Thus, disk diffusion susceptibilities which include these disks (especially polymyxin) will provide presumptive evidence of species as well as susceptibility data. PMID- 6298272 TI - Detection of genital herpes simplex infections by a tissue culture-fluorescent antibody technique with biotin-avidin. AB - Several cell lines were evaluated for their suitability for rapid detection of herpes simplex virus (HSV) from clinical genital specimens. Human foreskin fibroblast (Flow 7000) cells were found to be most suitable in terms of sensitivity and adherence characteristics. HSV in clinical specimens was isolated by a standard tissue culture method by monitoring the cytopathic effect, and the titers of the HSV-positive specimens were determined. More than 65% of the HSV positive genital specimens showed titers of less than or equal to 10(4) 50% tissue culture infective doses per ml. The standard tissue culture-cytopathic effect method required 3 to 10 days for detection of HSV in clinical specimens of low infectivity. A more rapid technique was developed which involved a short-term tissue culture (24 h) on Lab-Tek chambers followed by staining with biotin-linked HSV antibody and avidin-fluorescein conjugate. Because of the high binding affinity of this system due to multiple binding of biotin to avidin and multiple attachment of biotin to the antibody molecule, the biotin-avidin fluorescent antibody technique produced a quality of fluorescence far superior to that of the conventional fluorescent-antibody techniques. The tissue culture-biotin-avidin fluorescent-antibody method was as sensitive as the tissue culture-cytopathic effect test. This method provides an improved, more rapid test (26 h) for detecting HSV in clinical specimens. PMID- 6298273 TI - Epidemiology of rotavirus diarrhea in the Highlands of Papua, New Guinea, in 1979, as revealed by electrophoresis of genome RNA. AB - Results of gel electrophoresis of rotavirus genome RNA from feces of children in two provinces in Papua, New Guinea, suggest that the epidemiology of rotavirus infection in small communities with a total population of 3,000 may differ from that in urban or closely settled rural areas. PMID- 6298274 TI - Cephalexin-supplemented Jones-Kendrick charcoal agar for selective isolation of Bordetella pertussis: comparison with previously described media. AB - Four agar media, Jones-Kendrick (JK) charcoal agar unsupplemented, JK agar supplemented with 0.5 U of penicillin per ml, JK medium supplemented with 2.5 micrograms of methicillin per ml, and JK medium supplemented with 40 micrograms of cephalexin per ml, were evaluated to determine their ability to support growth of Bordetella pertussis, their ability to selectively inhibit normal pharyngeal flora while maintaining growth of B. pertussis, and their stability during storage. Five stock cultures of B. pertussis were plated on each of the media. Penicillin- and cephalexin-supplemented media were more inhibitory for early growth of B. pertussis than was medium supplemented with methicillin. However, after 5 days of incubation at 35 degrees C, all media supported good growth of this organism. When employed to detect B. pertussis in sham specimens, prepared by mixing normal pharyngeal material with each of the five B. pertussis stock cultures, the medium containing cephalexin was judged superior to all other media tested in its combined ability to suppress the growth of normal pharyngeal flora and to allow early detection of Bordetella colonies. All media tested retained their efficacy after 9 weeks of storage at 2 to 8 degrees C. PMID- 6298275 TI - Automatic and manual latex agglutination tests for measurement of cholera toxin and heat-labile enterotoxin of Escherichia coli. AB - Automated and manual latex agglutination methods were employed to measure cholera toxin (CT), heat-labile enterotoxin (LT) of Escherichia coli, and their subunits A and B. Dow polystyrene latex particles (diameter, 0.22 microns) and polystyrene chlorostyrene latex particles (diameter, 1 micron) were sensitized by rabbit specific immunoglobulin for each antigen and used as the reagents of the automated and manual agglutination tests, respectively. Automated agglutination was performed by a nephelometric assay system measuring time-dependent differences of light scattering due to agglutination, and manual latex agglutination was carried out in microtiter plates. As low as 1,000 and 31 pg of CT per ml were estimated by the automated and manual agglutination tests, respectively. Using these methods, the amount of CT and LT was measured in several clinical isolates of Vibrio cholerae and E. coli. Furthermore, it was discovered that cyclic AMP is not essential for the production of CT by measuring the amount of the toxin in numbers of cyclic AMP-dependent mutants of V. cholerae (with the agglutination tests). PMID- 6298276 TI - Typing of clinical herpes simplex virus isolates with mouse monoclonal antibodies to herpes simplex virus types 1 and 2: comparison with type-specific rabbit antisera and restriction endonuclease analysis of viral DNA. AB - A total of 122 clinical isolates of herpes simplex virus (HSV) from 107 patients were typed by using an indirect immunoperoxidase technique with commercially available type-specific rabbit antisera, recently developed mouse monoclonal antibodies to HSV types 1 and 2, and restriction endonuclease analysis of viral DNA. With the commercially available type-specific rabbit antisera, 34% of clinical HSV isolates were of indeterminate type; 63% of them were typed as HSV type 1 and 37% as HSV type 2 by using monoclonal antibody and restriction enzyme typing systems. Typing by immunofluorescence assay with the monoclonal antibodies gave identical results to those obtained by restriction enzyme analysis. Simultaneous infection with both HSV types was demonstrated by monoclonal antibody typing in five isolates from three patients. These findings were subsequently confirmed by plaque purification and restriction endonuclease analysis of viral DNA. Monoclonal antibodies were as sensitive as restriction enzyme analysis for the typing of clinical HSV isolates. Because of their simplicity, they are more amenable to use in clinical laboratories than is restriction endonuclease analysis. PMID- 6298277 TI - 2-Methylthioadenosine[beta-32P]diphosphate. An agonist and radioligand for the receptor that inhibits the accumulation of cyclic AMP in intact blood platelets. AB - 2-Methylthio-ADP and its radioactive analogue [beta-32P]2-methylthio-ADP were synthesized and used to investigate platelet receptors for ADP. 2-Methylthio-ADP induced platelet aggregation and shape change, and inhibited cyclic AMP accumulation in platelets exposed to prostaglandin E1. Compared with ADP, 2 methylthio-ADP was 3-5 times as active as an aggregating agent and 150-200 times as active as an inhibitor of cyclic AMP accumulation. Binding of [beta-32P]2 methylthio-ADP to platelets was measured after centrifuging them through silicone oil to separate platelets from their suspension medium. Binding was reversible, saturable, and specific, with between 400 and 1,200 sites/cell in different platelet preparations. There was no evidence for a second class of binding sites with different affinity. The second order association rate constant was approximately 3.5 X 10(6) M-1 S-1, and the first order dissociation rate was 0.024 s-1, both measured at 23 degrees C. The dissociation equilibrium constant (approximately 15 nM) was about three times higher than the concentration giving half-maximal inhibition of prostaglandin E1-stimulated cyclic AMP accumulation in platelet-rich plasma. Binding was inhibited by ADP (Ki = 3.5 microM), ATP (7 microM), 2-azido-ADP (0.12 microM), inosine diphosphate (IDP, 150 microM), guanosine diphosphate (GDP, 350 microM), and AMP (800 microM). Binding of 2 methylthio-ADP was also blocked by the non-cell-penetrating thiol reagent, p mercuribenzene sulphonate, a reagent that blocks the inhibition of adenylate cyclase by ADP, but which does not block the ability of ADP to induce aggregation or platelet shape change. The amount of 2-methylthio-ADP bound at saturation was independent of pH in the range 6-8, but the affinity was reduced at pH 6 compared with pH 6.5-8.0. The dissociation constant was not temperature dependent in the range 32 degrees -40 degrees C, whereas the rate of dissociation of 2-methylthio ADP from platelets after the addition of an excess of ADP approximately doubled over this range. The activation energy for dissociation was approximately 15 kcal/mol. Our results support the conclusion that platelets have a receptor for ADP, which inhibits cyclic AMP accumulation, and which has a sulphydryl group in the binding pocket. PMID- 6298278 TI - Adrenergic regulation of glycogenolysis in rat liver after cholestasis. Modulation of the balance between alpha 1 and beta 2 receptors. AB - The effects of extrahepatic cholestasis upon adrenergic regulation of glycogenolysis and upon the numbers of adrenoceptors in rat liver were studied using isolated hepatocytes and plasma membranes, respectively. A 60% decrease in the number of alpha 1 adrenoceptors (285 vs. 680 fmol/mg protein) and a simultaneous 2.7-fold increase in the number of beta adrenergic sites (67 vs. 25 fmol/mg protein) were observed beginning 36 h after bile flow obstruction and persisted for at least 68 h. The reciprocal modification of the numbers of alpha 1 and beta adrenoceptors was accompanied by a change in the manner of stimulation of glycogen phosphorylase by catecholamines in hepatocytes; originally alpha 1 adrenergic in normal rats (phenylephrine Ka = 0.9 microM, isoproterenol Ka = 7.1 microM), the stimulation became predominantly beta adrenergic in cholestatic animals (phenylephrine Ka = 3.7 microM, isoproterenol Ka = 0.06 microM). In normal rats, activation of the enzyme by epinephrine was inhibited by the alpha blocker phentolamine, without inhibition by the beta blocker propranolol. In contrast, propranolol was more effective than phentolamine in cholestatic rat hepatocytes. Modification of the regulation of glycogenolysis after cholestasis did not seem to be secondary to an alteration in the metabolism of thyroid hormones or in the action of glucocorticoids. However, cholestasis provoked a 10 fold increase in the number of hepatic mitoses and in the incorporation of thymidine into liver DNA of cholestatic animals. Similar changes were observed in regenerating livers, following two-thirds hepatectomy. We propose that the changes following extrahepatic cholestasis might, as well, be explained by a regenerative process. PMID- 6298280 TI - Effect of synthetic ovine corticotropin-releasing factor. Dose response of plasma adrenocorticotropin and cortisol. AB - Synthetic ovine corticotropin-releasing factor (CRF) was administered to normal male volunteer subjects as an intravenous bolus or 30-s infusion. Doses of CRF ranging from 0.001 to 30 micrograms/kg body wt were administered, and plasma immunoreactive (IR)-ACTH and IR-cortisol concentrations were measured. The threshold dose appeared to be 0.01-0.03 micrograms/kg, the half-maximal dose 0.3 1 micrograms/kg, and the maximally effective dose 3-10 micrograms/kg. Basal concentrations of IR-ACTH and IR-cortisol were 14 +/- 7.6 pg/ml (mean +/- SD) and 5.6 +/- 2.2 micrograms/dl, respectively. IR-ACTH rose as early as 2 min after CRF injection, reached peak levels in 10-15 min, and declined slowly thereafter. IR cortisol rose at 10 min or later and reached peak levels in 30-60 min. At a dose of 30 micrograms/kg, neither IR-ACTH nor IR-cortisol fell from peak levels of 82 +/- 21 pg/ml (mean +/- SE) and 23 +/- 1.4 micrograms/dl, respectively, during the 2-h course of the experiment, indicating that CRF has a sustained effect on ACTH release and/or a prolonged circulating plasma half-life. There was little or no increase in the levels of other anterior pituitary hormones. At doses of 1 microgram/kg and higher, facial flushing, tachycardia, and, in some subjects, a 15-29-mmHg decline in systemic arterial blood pressure were observed, even though blood volume was replaced and the subjects remained supine. These data indicate that synthetic ovine CRF is a very potent and specific ACTH secretagogue in man. Administered with caution until its vasomotor effects are more fully defined, CRF promises to be a safe and very useful investigative, diagnostic, and, possibly, therapeutic agent in man. PMID- 6298279 TI - In vitro desensitization of beta adrenergic receptors in human neutrophils. Attenuation by corticosteroids. AB - The receptor alterations involved in catecholamine-induced desensitization of adenylate cyclase in human neutrophils have been investigated as has the ability of hydrocortisone to modify such alterations. Incubation of human neutrophils with isoproterenol for 3 h in vitro resulted in an 86% reduction in the ability of isoproterenol to stimulate cyclic AMP accumulation in the cells. Two types of receptor alterations were documented. There was a 40% reduction in the number of beta adrenergic receptors (42 vs. 25 fmol/mg protein, P < 0.005) present after desensitization as assessed by [(3)H]dihydroalprenolol ([(3)H]DHA) binding. In addition the receptors appeared to be relatively uncoupled from adenylate cyclase. This uncoupling was assessed by examining the ability of the agonist isoproterenol to stabilize a high-affinity form of the receptor, detected by computer modelling of competition curves for [(3)H]DHA binding. Desensitized receptors were characterized by rightward-shifted agonist competition curves. When hydrocortisone was added to the desensitizing incubations (combined treatment) there was a statistically significant attenuation in the desensitization process as assessed by the ability of isoproterenol to increase cyclic AMP levels in the cells. Although combined treatment did not prevent the decline in receptor number, it did attenuate the uncoupling of the receptors. Combined treatment resulted in competition curves intermediate between the control and the rightward-shifted desensitization curves. Prednisolone was similar to hydrocortisone in attenuating isoproterenol-induced uncoupling. Thus, steroids appeared to attenuate agonist-induced desensitization of the beta adrenergic receptor-adenylate cyclase system by dampening the ability of agonists to uncouple receptors without modifying their ability to promote down-regulation of beta adrenergic receptors. PMID- 6298281 TI - Reduced platelet thromboxane formation in uremia. Evidence for a functional cyclooxygenase defect. AB - A qualitative platelet abnormality and a bleeding tendency are frequently associated with renal failure and uremia. We demonstrated previously that uremic patients display an abnormal platelet aggregation to arachidonic acid and reduced malondialdehyde production in response to thrombin and arachidonic acid. The objectives of this investigation were: (a) to compare platelet prostaglandin (PG) and thromboxane (TX) production in whole blood and in platelet-rich plasma (PRP) of 21 uremic patients and 22 healthy subjects; (b) to evaluate the concentration and activity of platelet PG- and TX-forming enzymes; (c) to assess the functional responsiveness of the platelet TXA(2)/PGH(2) receptor; (d) to explore the hemostatic consequences of partially reduced TXA(2) production.Platelet immunoreactive TXB(2) production during whole blood clotting was significantly reduced, by approximately 60%, in uremic patients as compared to age- and sex matched controls. Exogenous thrombin (5-30 IU/ml) failed to restore normal TXB(2) production in uremic platelets. Uremic PRP produced comparable or slightly higher amounts of TXB(2) than normal PRP at arachidonate concentrations 0.25-1 mM. However, when exposed to substrate concentrations >2 mM, uremic PRP produced significantly less TXB(2) than normal PRP. To discriminate between reduced arachidonic acid oxygenation and altered endoperoxide metabolism, the time course of immunoreactive TXB(2) and PGE(2) production was measured during whole blood clotting. The synthesis and release of both cyclooxygenase-derived products was slower and significantly reduced, at all time intervals considered. Furthermore, PGI(2) production in whole blood, as reflected by serum immunoreactive 6-keto PGF(1alpha) concentrations, was significantly reduced in uremic patients as compared with healthy subjects. PGH synthase levels, as determined by an immunoradiometric assay, were not significantly different in platelets from uremic patients as compared to control platelets. A single 40-mg dose of aspirin given to five healthy volunteers reduced their serum TXB(2) to levels found in uremic patients. This was associated with a significant increase of threshold aggregating concentrations of ADP and arachidonic acid and prolongation of bleeding time. Substantially similar threshold concentrations of U46619, a TXA(2) agonist, induced aggregation of normal and uremic platelets. Prostacyclin induced a significant elevation of uremic platelet cyclic AMP, which was suppressed by U46619, further suggesting normal responsiveness of the TXA(2)/PGH(2) receptor. WE CONCLUDE THAT: (a) an abnormality of platelet arachidonic acid metabolism exists in uremia, leading to a reduced TXA(2) production; (b) the characteristics of this abnormality are consistent with a functional cyclooxygenase defect; (c) reduced TXA(2) production may partially explain the previously described abnormality of platelet function in uremia. PMID- 6298282 TI - Human renal carcinoma cells produce hypercalcemia in the nude mouse and a novel protein recognized by parathyroid hormone receptors. AB - When grown in nude mice, cultured renal carcinoma cells from a hypercalcemic patient produced marked hypercalcemia that was reversed by resection of tumor. Conditioned medium from this cell line contained a protein with activity in a renal adenylate cyclase bioassay for parathyroid hormone (PTH) which was blocked by the competitive PTH antagonist [8norleucyl, 18norleucyl, 34tyrosinyl]bPTH (3 34)amide. However, the biologically active protein was eluted from gel filtration columns as a larger molecular size component that PTH and was not recognized by any of four region-specific PTH antisera. The properties of this factor resemble those of the postulated PTH-like substance(s) in humoral hypercalcemia of malignancy. PMID- 6298283 TI - Tyrosine crystals in a parotid pleomorphic adenoma in a Vietnamese boat person. PMID- 6298284 TI - Clostridium perfringens and other anaerobes isolated from bile. AB - Clostridium perfringens was isolated from bile in 13 cases of 150 patients examined. The serotypes of C perfringens strains isolated from bile and faeces were investigated using antisera to Hobbs' type 1-17. Two or more serological types were often found in a single specimen, but in the same patient the serotypes of C perfringens strains isolated from the bile were identical with those from the faeces. Beta-glucuronidase production in these C perfringens serotypes was tested with the API-Strep system. Strains agglutinated with Hobbs' antisera produced beta-glucuronidase, but non-agglutinated strains did not. PMID- 6298285 TI - Treatment of chromomycosis with ketoconazole and 5-fluorocytosine. AB - Chromomycosis is a disfiguring cutaneous mycosis that is often recalcitrant to therapy. We describe a case of chromomycosis of the lower extremities that is being successfully treated with ketoconazole and 5-fluorocytosine (5-FC) after treatment with ketoconazole alone had failed. In vitro studies showed that the drugs acted synergistically against the strain of Fonsecaea pedrosoi which was isolated from the patient's lesion. PMID- 6298287 TI - Arterioportal fistula in hepatocellular carcinoma. AB - A hepatic arterioportal fistula was visualized in a patient with hepatocellular carcinoma by sequential computed tomography (CT) after rapid bolus injection of contrast medium. Early contrast staining was demonstrated in the trunk and branches of the portal vein. The CT findings corresponded with those of celiac angiography. PMID- 6298286 TI - Nuclear magnetic resonance (NMR) imaging of Arnold-Chiari type I malformation with hydromyelia. AB - Saturation recovery nuclear magnetic resonance (NMR) images and metrizamide computed tomography (CT) scans were obtained in an adult patient with a clinical history suggestive of syringomyelia. Both NMR and CT studies showed low lying cerebellar tonsils. The CT study demonstrated central cavitation of the spinal cord from the midthoracic to midcervical levels but could not exclude an intramedullary soft tissue mass at the cervico-medullary junction. The NMR images in transverse, coronal, and sagittal planes demonstrated extension of an enlarged central spinal cord cerebrospinal fluid space to the cervico-medullary junction. This was felt to be strong evidence for exclusion of an intramedullary soft tissue mass and in favor of a diagnosis of Arnold-Chiari Type I malformation with hydromyelia. The noninvasive nature of spinal cord and cervico-medullary junction evaluation with NMR is emphasized. PMID- 6298288 TI - Pulmonary oil embolism after lymphangiography. PMID- 6298289 TI - Dynamic incremental computed tomography in evaluation of the pulmonary hila. AB - Contrast-enhanced dynamic computed tomography (CT) with table incrementation between scans was used to evaluate the hila in 100 patients without hilar abnormality and 15 patients with hilar pathology. This method of hilar evaluation was found to be superior to conventional CT because hilar vascular structures could be clearly opacified. Opacification of normal hilar vessels permitted accurate detection of hypovascular hilar masses, some of which were very small, nonlobular, and invisible using conventional CT. The technique was also found to be useful in evaluating the hila in patients with pulmonary artery dilatation or pulmonary opacification contiguous with hilar borders. We recommend that contrast enhanced dynamic incremental CT be used for hilar investigation in patients referred for hilar CT examination unless there is a specific contraindication to contrast medium administration. PMID- 6298290 TI - Computed tomography of calcified liver masses. AB - Our retrospective study of 230 computed tomographic (CT) studies of hepatic masses yielded 28 cases (12.2%) which contained calcification. Calcific deposits were noted in 9 of 59 patients (15.3%) with hepatocellular carcinoma (HCC), 15 of 82 patients (18.3%) with liver metastases from colorectal carcinoma (MCRC), 2 of 71 patients (2.8%) with noncolonic liver metastases, and 2 of 18 patients (11%) with a benign liver mass. The patterns of calcification in HCC and MCRC were highly variable and exhibited some overlap. Although the CT demonstration of calcification in HCC and MCRC is not uncommon, particularly in patients who have undergone radiotherapy or chemotherapy, we could identify no specific diagnostic pattern. PMID- 6298291 TI - CT demonstration and guided aspiration of unusual adrenal metastases. AB - Certain malignant neoplasms commonly metastasize to the adrenal glands, whereas others do so only infrequently. In the latter circumstance, an adrenal mass is likely to be benign. We report 3 patients with malignant neoplasms (hepatoma, transitional cell carcinoma of the bladder, and squamous cell carcinoma of the vulva) that do not commonly metastasize to the adrenals in whom computed tomographic (CT) demonstration and guided aspiration of adrenal masses significantly affected patient management. CT guided aspiration of adrenal masses is safe and accurate and is recommended whenever a diagnosis of adrenal metastasis would alter the staging or therapy of the primary neoplasm. PMID- 6298292 TI - Effect of environmental heat and dietary fiber on thyroid physiology of lactating cows. AB - Six lactating dairy cows were assigned randomly to a diet either of low fiber (30% corn silage) or of high fiber (70% corn silage). Both groups were subjected to 10 days thermoneutral (17.6 degrees C) and 10 days of environmental heat (31.2 degrees C) to determine effects of dietary fiber content and environmental heat on concentrations of thyroxine and triiodothyronine in plasma, milk, urine, and feces. During heat there were significant reductions in concentrations of thyroxine and triiodothyronine in plasma and in excretion of thyroxine and triiodothyronine in milk. Excretion of thyroxine and triiodothyronine increased in urine during heat stress, whereas concentration and excretion of thyroxine and triiodothyronine in feces decreased. Dietary fiber content had little effect upon thyroxine and triiodothyronine concentrations in plasma or excretion in milk, feces, and urine. Excretion of triiodothyronine in milk was negatively correlated with rectal temperature and positively with content in plasma. The reduction in thyroxine and triiodothyronine of plasma and the decline in excretion of thyroxine and triiodothyronine suggested that environmental heat reduced synthesis of both hormones. PMID- 6298293 TI - Parasuicide. A review of treatment interventions. AB - From review of studies of treatment intervention in parasuicide it is concluded that: (1) Suicide prevention centres do not lower the incidence of suicide. (2) Retrospective follow-up studies of patients self-selected for treatment or no treatment find that patients who attend for treatment have a lower repeat rate of parasuicide. This may have nothing to do with treatment. (3) Intensive domicillary follow-up for 3-6 months after inpatient treatment in a specialised unit for parasuicide is no better at preventing further parasuicides than conventional follow-up in outpatient department or via the GP. (4) A multidisciplinary outreach programme was effective in one study but not three others. (5) Behaviour therapy was no better than insight oriented psychotherapies when administered intensely over 10 days for inpatients with a previous history of overdose. (6) Medication in the form of depot flupenthixol, 20 mg every 4 weeks was significantly better than placebo for chronic repeaters (3 or more attempts) but Mianserin 30 mg/d was not. (7) There was no relationship between improved symptoms, improved social circumstances or the repeat rate for parasuicide. In a controlled trial conducted by the authors in parasuicides, Mianserin 60 mg/d reduced depressive symptoms significantly more rapidly than Nomifensine or placebo but there was no difference by 6 weeks and the risk of repeat was not affected. PMID- 6298294 TI - The NIMH diagnostic interview schedule modified to record current mental status. AB - The National Institute of Mental Health Diagnostic Interview Schedule (DIS) was modified to record detailed information on current mental status in addition to the lifetime symptom history. Use of the modified DIS in a field survey indicates that information on current symptoms is required to distinguish persons who meet all DSM-III criteria for Major Depressive Episode at or around the time of interview from former cases who fail to meet all criteria at interview. Thus, the unmodified DIS may overestimate the 1-month period prevalence rates for Major Depressive Episode, by counting symptomatic former cases as having the disorder at or around the time of the interview. An analysis of symptom count data also suggests that the unmodified DIS count of lifetime depressive symptoms is not a good measure of current symptom status. PMID- 6298295 TI - Recent advances in knowledge about beta-adrenergic receptors: application to clinical cardiology. AB - The properties of beta-adrenergic receptors in the cardiovascular system have been studied in the past by two experimental approaches, which can be termed pharmacologic and biochemical. In the pharmacologic approach, the nature of a drug interaction with receptors is deduced from alterations in the physiologic properties of the tissue caused by administration of various concentrations of the drug. Many important concepts about beta-adrenergic receptors have come from such indirect pharmacologic studies. The biochemical approach directly assesses the interaction of drugs with beta-adrenergic receptors by studying the binding of radiolabeled antagonists and agonists with the receptor. This relatively new approach has provided a large amount of new information regarding the intrinsic properties of beta-adrenergic receptors and modification of these properties by physiologic stresses, administration of drugs and disease states. The biochemical approach has also been applied recently to the study of beta-adrenergic receptors in human beings. In the future, substantial clinically relevant new information regarding the nature of beta-adrenergic receptors in physiologic and pathologic conditions should result from application of a combination of the biochemical and physiologic approaches to studies in human beings. PMID- 6298296 TI - Relative value of isotonic and isometric exercise radionuclide angiography to detect coronary heart disease. PMID- 6298297 TI - Global and regional left ventricular ejection fraction abnormalities during exercise in patients with silent myocardial ischemia. AB - Sixteen asymptomatic patients with coronary artery disease and silent myocardial ischemia were studied with exercise radionuclide ventriculography. Radionuclide ventriculograms were analyzed for changes in ejection fraction globally and in three regions. Results were compared with radionuclide ventriculograms in 24 symptomatic patients. Both groups (silent myocardial ischemia and angina) were similar in prevalence of multivessel disease and previous myocardial infarction, as well as in age and sex. Global ejection fraction decreased by 0.06 in both groups during exercise; regional ejection fraction also decreased by similar amounts in the two groups. Furthermore, the percent of regions with normal ejection fraction at rest that demonstrated a decrease during exercise was identical: 19 (60%) of 33 versus 26 (60%) of 46. These exercise radionuclide ventriculographic results suggest that abnormalities in regional and global left ventricular wall motion are similar in patients with coronary artery disease with and without silent myocardial ischemia. PMID- 6298298 TI - Timed effects of an oral adrenergic bronchodilator on pulmonary function and measurements of plasma and leukocyte cyclic AMP: divergence in duration of drug action and development of tolerance. AB - The time course of response to initiation and withdrawal of treatment with terbutaline (5 mg orally, three times daily) was examined in eight normal subjects and 14 asthmatic patients. The various indices of response, examined simultaneously, yielded divergent estimates of the duration of drug action and the development of tolerance during continuous treatment. Bronchodilator responses were observed after each dose of terbutaline, but within 8 hr after a dose, pulmonary functions were similar to those observed in the absence of treatment. There was a suggestion of moderate tolerance to the bronchodilator effects of medication in that areas under the curve for increments in pulmonary function after terbutaline were lower during treatment than after the initial doses of medication; but the variations in magnitude of pulmonary response on different study days did not achieve statistical significance. Measurements of plasma cyclic AMP concentration suggested a much longer duration of drug action, with elevated levels observed 24 hr after a treatment dose; there was a progressive decrease in the plasma cyclic AMP responses to medication during the course of treatment. Conversely, leukocyte cyclic AMP responses to adrenergic stimulation were suppressed within 4 hr after the first dose of medication; the suppression persisted throughout treatment and for 1 to 3 days thereafter. These observations indicated that duration of drug action and induction of tolerance vary in different organ systems. PMID- 6298299 TI - The trace metal status of a group of post-menopausal vegetarians. AB - The copper, manganese, and selenium status of 36 post-menopausal vegetarians was compared with that of a group of post-menopausal omnivores, using dietary trace element intake data, serum and hair copper levels, and hair manganese and hair selenium levels as the principal indexes. Findings of this study suggest that the copper and selenium status of these long-term vegetarian women was comparable to that of non-vegetarians, despite the high intake of dietary fiber by the vegetarian group. In contrast, the manganese status of the vegetarians, as indicated by elevated hair manganese levels, was higher, almost certainly as a result of the significantly higher manganese intake of this group. PMID- 6298300 TI - Planning high-carbohydrate, high-fiber diets with a microcomputer. AB - Current research suggests that a diet high in carbohydrate and fiber (HCF) may be useful in the management of diabetes mellitus, hypercholesterolemia, and hypertriglyceridemia. To meet the needs of our clinic population, a microcomputer program has been developed that allows for individualization of HCF menu patterns that satisfy both the diet prescription and the patient's preferences. Educational materials were prepared to guide and augment the HCF diet instruction. It is hoped that this comprehensive approach to diet planning and teaching can be applied in similar settings to facilitate the delivery of quality patient care. PMID- 6298301 TI - Aging of the rat adrenocortical cell: response to ACTH and cyclic AMP in vitro. AB - To study intrinsic age-related changes in adrenocortical steroid production, cells isolated from rats of different ages (3 to 24 months) were used. Acute (2 hour) corticosterone production in response to stimulation by adrenocorticotrophic hormone (ACTH) and adenosine 3':5'-cyclic monophosphate (cAMP) was measured by radioimmunoassay. With age, adrenocortical cells lose much of their ability to produce corticosterone in the absence or presence of ACTH or cAMP. The loss is progressive from 6 to 24 months of age. Analysis of the data suggests that from 6 to 12 months, an intracellular steroidogenic lesion develops; in addition there may be a loss in ACTH receptors on the plasma membrane. After 12 months these defects increase and are accompanied by a decrease in receptor sensitivity to ACTH. PMID- 6298302 TI - Decreased beta-adrenergic agonist affinity and adenylate cyclase activity in senescent rat lung. AB - Beta-adrenergic receptor affinity for the agonist, isoproterenol, and coupling to adenylate cyclase assessed by Hill plots were investigated at both 23 degrees C and 37 degrees C in Fischer 344 rats of 4, 13, and 25 months of age. Apparent dissociation constants for isoproterenol at 23 degrees C were unaltered in the three age groups (2.0 +/- .3; 2.2 +/- .3; 2.6 +/- .3 X 10(-7)M, respectively). At 37 degrees C, however, the apparent dissociation constants increased progressively with age (6.9 +/- .9;8.6 +/- .6;13.2 +/- 1.4 X 10(-7)M). Hill coefficients were less than 0.7 for all age groups, suggesting the presence of two binding sites that have been described as a low (uncoupled) and a high (coupled) affinity state. In the presence of guanylylimidodiphosphate the apparent dissociation constant increased in all age groups and there was no longer a difference among age groups at 37 degrees C (14.1 +/- 2.7; 13.6 +/- 1.7; 14.8 +/- 2.4 X 10(-7)M). Basal adenylate cyclase activity (17 +/- 1 vs. 25 +/- 3 pmols cAMP/mg/min) as well as isoproterenol (27 +/- 5 vs. 60 +/- 11), NaF (116 +/ 8 vs. 209 +/- 24), and forskolin (70 +/- 6 vs. 124 +/- 14) stimulated activity were reduced in 25 compared to 4-month-old animals. The data suggest that in lungs from senescent rats there is a decrease in beta-adrenergic agonist affinity of the high affinity binding site and no change in the affinity of the low affinity binding site. Additionally, adenylate cyclase activity is reduced in senescent compared with young rats, and this reduction may be due to decreased activity of the catalytic unit. Both these changes may contribute to the diminished hormonal responsiveness of senescence. PMID- 6298303 TI - [Paget's disease of the vulva associated with Paget's disease of the breast. Apropos of a case report. Review of the literature]. AB - It is rare to find associated Paget's disease of the vulva and Paget's disease of the nipple (fewer than 100 cases have been published). A case history with this association in a woman of 72 years of age is reported here. Although the diagnosis of Paget's disease was not made at the same time in the two sites, the treatment was carried out at the same time. This case is, as far as we know, a unique case because the vulval Paget's disease was not accompanied by an infiltrating underlying carcinoma, whereas the nipple Paget condition was associated with a non-invasive cancer of the breast. After reviewing the literature the commentators have developed a clinical and anatomo-pathological plan. A very exact anatomo-pathological study was carried out with a search for oestradiol receptors and these contribute to the establishment of the link between these two pathological states and a way of understanding their pathogenicity better. PMID- 6298304 TI - The histopathologic and clinical indicators of prognosis in hepatoma. AB - Two histopathologic subtypes of hepatoma, clear cell type and fibrolamellar type, have been reported to indicate a longer survival. Although data on the prognostic value of clear cell histology is equivocal, evidence for prolonged survival (mean survival: 32-68 months) for patients with fibrolamellar type is impressive. Aggressive surgical intervention, including resection of metastases, appears indicated in fibrolamellar hepatocellular carcinoma. Bilirubin determination may be a reliable indicator of survival, but conflicting results are reported for most reputed clinical prognostic markers. Discrepancies may reflect regional and ethnic differences in the pathogenesis of hepatoma. We present an illustrative case of fibrolamellar hepatoma discovered in a 24-year-old woman with migratory thrombophlebitis. The patient successfully underwent an extended right hepatic lobectomy and is currently free of disease. We review the histopathologic and clinical prognostic features of fibrolamellar carcinoma and hepatoma. PMID- 6298305 TI - Molecular cytochemistry of the replication and transcription of chromatin. PMID- 6298306 TI - Membrane Na/K-adenosine triphosphatase (ATPase) (K-P-nitrophenylphosphate) in epithelial cells. PMID- 6298307 TI - Corticotropin (ACTH) and the N-terminal fragment of pro-opiomelanocortin are located in the same granules in cells of rat pituitary gland. AB - The localization of corticotropin (ACTH) and of the N-terminal fragment (NTF) of pro-opiomelanocortin were assessed by light and electron microscope immunochemistry. Using the unlabeled technique of Sternberger at the light microscope level, a strongly positive reaction for both NTF and ACTH antisera was observed in all secretory cells of the pars intermedia. In the pars distalis, immunostaining with ACTH antiserum was localized in stellate cells dispersed throughout the lobe. As observed in serial, consecutive sections, the NTF antiserum stained exactly the same cells. At the electron microscope level, using the protein A-gold technique, all the secretory granules of the cells of the pars intermedia showed a positive reaction with both antisera. In the pars distalis, after exposure to either of the antisera, gold particles were found over secretory granules of typical stellate corticotrophs. Making use of the possibility of reacting both faces of a fine section with gold particles of different sizes, it was found that the same secretory granules in all the cells of the pars intermedia and in corticotrophs of the pars distalis contained particles of both sizes. These results indicate that ACTH and NTF are contained in the same secretory granules and released together in the circulation where NTF is found in high amounts. PMID- 6298308 TI - Immunohistochemical localization of collagenase in hepatic murine schistosomiasis. AB - It has been previously demonstrated that collagenase activity and collagen synthesis in hepatic granulomas of mice infected with S. mansoni cercariae are maximal 8 weeks after infection; however, total liver collagen content continues to increase. Now the anatomic relationships among collagenase and collagen, granulomas, and hepatic parenchyma in normal mice and in mice infected with S. mansoni are studied. Trypsin-activated collagenase was purified from the media of cultured granuloma explants and anti-collagenase immunoglobulin G was purified from immunized rabbits. The IgG cross-reacted with liver granulomas and active and inactive forms of collagenase, but did not react by immunodiffusion in agar with other neutral proteases or homogenates of schistosome eggs or normal liver. Cryostat sections of liver from normal and infected mice were studied by indirect immunohistochemical methods using fluorescein, rhodamine, and peroxidase labels. Collagenase localization was restricted to areas of collagen deposits in granulomas and hepatic parenchyma. Ultrastructural studies revealed collagenase on the surface of collagen fibers. Hepatocytes of normal mice showed delicate staining at the sinusoidal surface. At all times, immunoreactive collagenase was intimately associated with its substrate, where it presumably initiated collagen degradation. This localization provides a rationale for possible therapeutic approaches to control fibrogenesis through collagenase induction or activation. PMID- 6298309 TI - Selective digestion of human metaphase chromosomes by Alu I restriction endonuclease. AB - Human cytological preparations have been digested with Alu I restriction endonuclease (Endo R Alu I) and subsequently stained with acridine orange. Metaphase chromosomes treated in this way revealed a clear-cut bright/ dull longitudinal differentiation. It has been postulated that these cytological findings could be related to the selective digestion, by Alu I, of all DNA with the exception of some heterochromatic, possibly satellite, fractions. PMID- 6298311 TI - A single centrifugation step method for the simultaneous separation of different leukocytes with special reference to basophilic leukocytes. AB - A simple method of separating mononuclear leukocytes, basophils, neutrophils and an eosinophil fraction from human blood is described, using discontinuous Percoll gradient centrifugation. Mononuclear cells were found in the upper layer (band 1) at a density of less than or equal to 1.072. The second band with a density of less than or equal to 1.082 contained most of the basophilic leukocytes (av. 70% and 8-20% of the differential cell count) and a minor part of the mononuclear cells (av. 8%). In the third band, density less than or equal to 1.1, 82-91% of the polymorphonuclear cells (PMN) were found, consisting mainly of neutrophils. 35-70% of the eosinophilic PMN cells are located in band four (density greater than 1.1). The function of the basophil-enriched fraction was tested by degranulation with specific antigen. PMID- 6298310 TI - A radioimmunoprecipitation polyethylene glycol assay for circulating Entamoeba histolytica antigens. PMID- 6298312 TI - Affinity purification of antibodies of regulatory subunits of cAMP-dependent protein kinase using cross-linked immunoabsorbent. PMID- 6298313 TI - Ly- and ecotropic MuLV antigens are separate entities. AB - Murine alloantisera to lymphocyte (Ly) alloantigens also contain antibodies to murine leukaemia viruses (MuLV) in high titre. A number of such sera were extensively absorbed with ecotropic AKR murine leukaemia virus to remove contaminating anti-MuLV antibodies. The virus-absorbed and unabsorbed antisera were then examined by the radioimmune precipitation (RIP) assay for anti-MuLV antibodies, and for anti-Ly antibodies by cytotoxicity assays against specific target cells. The results presented here demonstrate that whilst virus absorption markedly reduced the anti-viral titres of the alloantisera they had no effect on the anti-Ly titres of any of them. This indicates that none of the Ly antigens which was examined cross reacts with the antigens of ecotropic AKR-MuLV and further suggests that Ly antigens are not encoded by the genes of endogenous ecotropic MuLVs. It was also demonstrated that the majority of the contaminating anti-MuLV antibodies present in anti-Ly-6.2 antiserum recognize viral envelope antigens rather than the internal structural proteins of the virus. PMID- 6298314 TI - Keratoconjunctivitis associated with adenovirus type 37: an extended outbreak in an ophthalmologist's office. AB - An outbreak of epidemic keratoconjunctivitis occurred in three clusters among patients of an ophthalmologist during 1977-1978. Cases in the third cluster and possibly one case in the first were caused by infection with adenovirus type 37. Among 83 infected patients, the mean interval between a visit to the ophthalmologist's office and the onset of symptoms was 11 days. Conjunctival specimens from 64 (77%) of the patients yielded adenovirus type 37, and adenovirus hexon antigen was identified by counterimmunoelectrophoresis in pooled specimens from the office environment. Infected patients were older and had more glaucoma, and were therefore tested more frequently by tonometry, than noninfected control patients. The association of this outbreak with adenovirus type 37 was made in 1981, the year in which this new serotype was fully described and reference reagents were produced. This is the first report of infection due to adenovirus type 37 in the United States. PMID- 6298315 TI - Serologic diagnosis of pertussis: comparison of enzyme-linked immunosorbent assay and bacterial agglutination. AB - An enzyme-linked immunosorbent assay (ELISA) and bacterial agglutination (BA) method for determining the presence of antibodies to Bordetella pertussis were compared on serum samples from 21 patients with whooping cough and their 76 family members. The overall diagnostic agreement between the two methods was 77%. The data for BA-detected antibodies correlated best with IgG and IgA antibodies to B. pertussis. All of the culture-positive patients showed serologic positivity in both assays during the follow-up. Pertussis was diagnosed by ELISA in most cases from the first serum sample. Both methods proved to be good diagnostic aids in culture-negative patients, although the value of BA is more retrospective because of the need for paired sera. The kinetics of IgM, IgA, IgG, and agglutinating antibodies to B. pertussis is presented. PMID- 6298316 TI - Production of coproantibodies and immune complexes in piglets infected with rotavirus. AB - The local immune response of newborn piglets to experimental infection with rotavirus was analyzed by means of coproantibodies, immune complexes, and the detection of rotavirus. Immune complexes appeared in the feces on day 4 after inoculation when the production of rotavirus began to decrease; they reached a peak when no rotavirus could be detected in the feces. The presence of free IgA and IgM antibodies to rotavirus could be demonstrated on day 7 when the immune complexes had already reached a maximum level. An enzyme-linked immunosorbent assay was used for all of the analyses. PMID- 6298317 TI - Sendai virus infection in the brains of mice: distribution of viral antigens studied with monoclonal antibodies. AB - Newborn and 12- and 21-day-old mice were inoculated intracerebrally with Sendai virus. Titers of infectious virus peaked on days 1-2 after infection and had disappeared after three days in 12- and 21-day-old mice and six days in newborn mice. The levels of infectious virus in the brain declined before the appearance of serum antibodies, which in newborn mice was delayed until day 12. Immunofluorescence analysis showed viral antigens in neurons and their dendritic processes 12 and 24 days after infection in newborn animals but not in older ones. Immunofluorescent staining with monoclonal antibodies to five major structural components of Sendai virus--the nucleocapsid, polymerase, matrix, fusion, and hemagglutinin-neuraminidase proteins--showed that all were present in choroid plexus epithelial cells and ependymal cells during acute infection. However, during acute and persistent infections in neurons, the first three antigens were found, but the last two--surface glycoproteins--were lacking or found at low levels. PMID- 6298319 TI - [Application of enzyme-linked protein A to ELISA for detection of virus specific antibodies]. PMID- 6298320 TI - [Seroepidemic survey of Coxsackie B viruses in Gifu prefecture, Japan, 1960 1980]. PMID- 6298318 TI - Topically administered acyclovir in the treatment of recurrent herpes simplex genitalis: a controlled trial. AB - Eighty-eight patients with culture-proven recurrent herpes simplex genitalis were entered into a collaborative, randomized, placebo-controlled, double-blind trial to evaluate the efficacy and toxicity of a topical formulation of acyclovir. Patients entered the study within 48 hr of the onset of lesions, and the study medication was applied six times daily for five days. The duration of virus shedding from lesions present at the time of entry into the study was significantly reduced for men who received acyclovir compared with men who received placebo (P less than 0.05). There were no significant differences between the acyclovir- and placebo-treated groups of either sex in time to crusting of lesions, time required for lesions to heal, time to cessation of pain, or in frequency with which new lesions developed during the course of therapy. Mild, transient burning or pain associated with application of the study medication was a common complaint. PMID- 6298321 TI - [Hemodynamic effects of dibutyryl cyclic AMP in patients following open-heart surgery]. PMID- 6298322 TI - [Dose response and duration of the effects of DBcAMP on hemodynamics and metabolism]. PMID- 6298323 TI - [Prediction of postoperative respiratory function of lung cancer patients using quantitative lung scans]. PMID- 6298324 TI - [Studies on the amino acid transport systems in the human placenta--L-alanine and L-leucine uptake by microvillous brush border membrane vesicles]. AB - To characterize the placental amino acid transport systems, L-alanine and L leucine uptakes were studied using microvillous brush border membrane vesicles prepared from human placenta. The specific activities of alkaline phosphatase and 5'-nucleotidase in the membrane preparation were enriched 9-11 times as high as those in the homogenate. Intravesicular water (IVW) volume determined with 3-O methyl-D-glucose was 0.59 microliters/mg protein. The saturation kinetics of L leucine uptake by the vesicles equilibrated with Na+ gave a single set of Km (4.2mM) and Vmax (1.16 mumol/ml IVW/30s). These parameters were clearly different from those for L-alanine uptake reported previously (Asai et al.: Biochem. Int., 4:377, 1982). In the presence of an inward Na+-gradient L-leucine uptake was stimulated about 2 times, but transient accumulation was not observed differing from L-alanine uptake. Discrimination of the neutral amino acid transport systems in the presence of an inward 100mM Na+-gradient revealed that the relative contributions of A, ASC and L systems, and simple diffusion were 55, 20, 15 and 10% for L-alanine, and 45, 0, 15 and 40% for L-leucine, respectively. The results indicate that the neutral amino acid transport systems in the human placental microvillous membranes are clearly different between L-alanine and L-leucine. PMID- 6298325 TI - [Evaluation of preoperative transcatheter arterial embolization for hepatocellular carcinoma]. PMID- 6298326 TI - [Evaluation of intratumoral injection of an immunopotentiator (OK-432) in patients with hepatocellular carcinoma]. PMID- 6298327 TI - Indirect hemagglutination inhibition: a direct method for detecting cytomegalovirus antigen. AB - We have developed a simple and sensitive technique for detecting CMV antigen in tissue culture specimens. The IHI assay is a modification of the IHA, which is capable of reliably detecting 10(3) median TCID50 of virus. With the IHI assay, antigen is detected by its capacity to inhibit the reactivity of CMV antisera of known titer in an IHA assay. The test is specific, uses readily available reagents, and is easy to perform. The IHI assay may serve as a tool for screening tissue culture specimens for CMV. PMID- 6298328 TI - The effect of gentamicin on antidiuretic hormone-stimulated osmotic water flow in the toad urinary bladder. AB - Early gentamicin nephrotoxicity is characterized by a variety of renal transport abnormalities, including polyuria secondary to nephrogenic diabetes insipidus. To investigate whether gentamicin directly interferes with cellular mechanisms responsible for ADH responsivity as a possible contributing factor in this tubular transport alteration, the effect of gentamicin on ADH-stimulated water flow was examined in vitro in the toad urinary bladder. Gentamicin decreased ADH stimulated osmotic water flow in a dose-dependent manner, with a threshold effect at approximately 0.5 mM. This inhibitory effect occurred only in response to submaximal concentrations of ADH, was reversible, and was preventable with Mg++. Additionally, gentamicin reduced theophylline-stimulated osmotic water flow but had no effect on cyclic AMP-induced water flow. These effects are consistent with a direct gentamicin-related decrease in ADH-responsive adenylate cyclase activity and suggest a mechanism that may contribute to the nephrogenic diabetes insipidus characteristic of early gentamicin nephrotoxicity and the nonoliguric acute renal failure that is a late toxic event occurring with the use of this antibiotic. PMID- 6298329 TI - C. T. evaluation of a peripheral arterio-venous malformation. PMID- 6298330 TI - Pain-inducing laryngeal paragangliomas (non-chromaffin). PMID- 6298331 TI - Intraductal carcinoma of the minor salivary gland. PMID- 6298332 TI - [Congenital angiodysplasias of types F.P. Weber, Klippel-Trenaunay and Servelle Martorell]. AB - Basing our observations on a study of 47 cases of angiodysplasia of types F.P. Weber, Klippel-Trenaunay and Servelle-Martorell, we attempted to demonstrate that differentiation of these 3 syndromes is possible if criteria obtained from a non invasive investigative method are used. This involves taking standard X-rays of the extremities (both sides) which are examined under direct magnification (0.1 01 mm) thus allowing the most exact possible analysis of the skeletal changes. In this way, the Weber syndrome should be suspected if bone prolongation is seen in association with loss of substances from the skeleton. In the Klippel-Trenaunay syndrome, the bone lengthening is not accompanied by bony lesions. In the Servelle-Martorell syndrome bony lesions go hand in hand with limb hypertrophy. PMID- 6298333 TI - Malignant fibrous histiocytoma (MFH) of the lung. A case report. PMID- 6298334 TI - Oestrogen metabolism in the endometrium, corpus luteum and ovarian residual tissue of the rabbit. AB - Reproductive tissues (uterine endometrium, corpus luteum and ovarian residual tissues) from pregnant and pseudopregnant rabbits were incubated with equimolar concentrations of [3H]oestrone and [3H]oestrone sulphate (0.375 pmol) to monitor the changes in oestrogen metabolism during the early stages of pregnancy (days 0, and 3-8 post coitum) and to investigate the embyonic effect upon maternal oestrogen metabolism. Oestradiol-17 beta was the major metabolite formed from oestrone and sulphoconjugation occurred in all tissues studied. Oestrone sulphate was converted primarily to oestradiol-17 beta-3-monosulphate. Endometrial 17 beta oxidoreductase significantly decreased and sulphotransferase increased in activity during the preimplantation period, but no differences were noted between gravid and non-gravid horns in unilaterally pregnant animals, nor between pregnant or pseudopregnant animals. Significant decreases occurred in 17 beta oxidoreductase and sulphotransferase activity in luteal tissue, but these were more than offset by increases in tissue weight. No differences in the activities in luteal tissues were detected between pregnant or pseudopregnant animals, nor between ovarian tissue adjacent to gravid or non-gravid uterine horns. The results show that significant changes occur in oestrogen metabolism in the rabbit endometrium and corpus luteum within 8 days after ovulation, and that these changes result from maternal factors expressed systemically rather than by the effects of the developing conceptus expressed locally. PMID- 6298336 TI - Receptors for insulin and insulin-like growth factor in cultured arterial smooth muscle cells depend on their growth state. AB - The binding of 125I-labelled insulin and 125I-labelled insulin-like growth factor (IGF) to cultured arterial smooth muscle cells from rats was studied during various growth states of the cells. The level of binding of 125I-labelled insulin to the cells was low in growing cells and high in stationary cells. The level of 125I-labelled IGF binding to the cells was high in growing cells and low in stationary cells. In addition, the effect of unlabelled IGF and insulin on the binding of both 125I-labelled hormones to the cells was examined during various growth states. In growing cells insulin displaced 125I-labelled insulin from its binding sites; IGF competed weakly with 125I-labelled insulin for the binding sites. In parallel, IGF displaced 125I-labelled IGF binding whereas insulin competed weakly with 125I-labelled IGF for the binding sites. In stationary cells both hormones displaced 125I-labelled IGF binding. Insulin-like growth factor also displaced 125I-labelled insulin binding whereas insulin could not significantly displace 125I-labelled insulin from the binding sites. Insulin only competed with 125I-labelled insulin for the binding sites after removal of the fetal calf serum from the culture medium. PMID- 6298335 TI - Thyrotrophin and the differential expression of proliferation and differentiation in dog thyroid cells in primary culture. AB - Cell proliferation and the expression of differentiated functions are generally considered to be mutually exclusive states of the cell. Thyrotrophin activates the expression of differentiated functions in dog thyroid cells by means of cyclic AMP. We have recently shown that thyrotrophin also acts through the same intracellular signal molecule to enhance proliferation of dog thyroid cells in primary cultures. In this work we showed that such primary cultures exhibit three successive phases: a latency period during which the expression of differentiated functions (iodide trapping and organification) declined, the cells still being associated with structures derived from the seeded follicles; a cell proliferation phase with little expression of these functions, the cells being spread in a monolayer; a stationary phase with cell density reaching a plateau but no re-expression of the functions. Thyrotrophin promoted proliferation during the multiplication phase, but induced redifferentiation during the stationary phase. These effects were mimicked by cholera toxin and dibutyryl cyclic AMP, which suggested that they were mediated by cyclic AMP. Iodide uptake was also stimulated by cortisol. Thyrotrophin therefore has a different action in dog thyroid cells depending on the state of the cells. The spontaneous arrest of multiplication appears to make the cells competent to respond to thyrotrophin by the induction of redifferentiation. PMID- 6298337 TI - The corticotrophic cells of the canine pituitary gland in pituitary-dependent hyperadrenocorticism. AB - The distribution of specifically stained corticotrophic cells has been studied in the pituitary glands of 11 dogs with pituitary-dependent hyperadrenocorticism. The results suggest that the disease is not a single entity, and that some cases are caused by primary abnormality of the pituitary gland whereas others appear to be the result of dysfunction of the hypothalamus or central nervous system. The patterns correspond closely to those demonstrated in the human pituitary gland in Cushing's disease, and confirm that the canine disease is a useful model for the study of the pathogenesis of the variants of the condition. PMID- 6298338 TI - Effects of divalent cations on 5'-nucleotidase activity of Formosan cobra venom. PMID- 6298339 TI - Anti-HBs reactivity in hepatitis B surface antigen positive serum samples--with special emphasis on heterotypic antibody. PMID- 6298340 TI - Echographic pattern of hepatocellular carcinoma. PMID- 6298341 TI - [A case of melanotic neuroectodermal tumor of infancy]. PMID- 6298342 TI - Prostaglandins enhance intercellular adhesion of Vero cells infected with herpes simplex virus. AB - The effect of prostaglandins (PGs) and cyclic nucleotides on adherence between uninfected Vero cells or between uninfected cells and cells infected with wild type or syncytial strains of herpes simplex virus (HSV) was studied. In the absence of serum, PGE2 increased adhesion between uninfected cells whereas PGF2 alpha decreased it. However, both PGE2 and PGF2 alpha increased adhesion between cells infected with HSV and uninfected cells. Moreover, dibutyryl cyclic AMP increased adhesion between uninfected cells and cells infected with HSV, whereas dibutyryl cyclic GMP decreased it. PMID- 6298343 TI - Ribonucleotide reductase induced by herpes simplex virus has a virus-specified constituent. AB - Ribonucleotide reductase, an enzyme found in all prokaryotic and eukaryotic cells that synthesize DNA, is induced by herpes simplex virus (HSV). In this study the effect of anti-HSV antiserum on the induced ribonucleotide reductase has been examined and the ability of different temperature-sensitive (ts) mutants of HSV-1 to induce the enzyme has been investigated. The HSV-1-induced ribonucleotide reductase was inhibited by antiserum raised against infected cell lysates but not by preimmune serum. The wild-type (ts+) virus induced similar levels of ribonucleotide reductase at 31 degrees C and 38.5 degrees C (the permissive and non-permissive temperatures respectively for the ts mutants). All ts mutants induced approximately wild-type levels of the enzyme at 31 degrees C. At 38.5 degrees C, two of the four ts mutants studied also induced wild-type levels of enzyme but ts G failed to induce any activity while ts K induced variable but low levels. The enzyme activity induced by ts G at 31 degrees C was thermolabile both in vivo and in vitro. These results provide the first strong evidence that the induced ribonucleotide reductase activity is at least partially virus-coded. PMID- 6298344 TI - Characterization of abnormal thymidine kinases induced by drug-resistant strains of herpes simplex virus type 1. AB - Two TK+ acyclovir-resistant variants of herpes simplex virus (HSV) (S1 and Tr7) and one TK+ BVdU-resistant variant (B3) induce abnormal thymidine kinases with impaired ability to phosphorylate the drugs used in their isolation. These enzymes have been purified and their properties compared with those of the wild type (wt) parent, SC16. The enzyme induced by S1 differed markedly from the other three in both its responses to salt and to pH. B3 TK recognized the enzyme's natural substrates, thymidine, deoxycytidine, dTMP and ATP as well as the wt enzyme. In contrast, Tr7 and S1 TKs failed to bind deoxycytidine and bind thymidine less well than wt. Tr7 and S1 TKs had affinities for dTMP similar to those of B3 and the wt enzymes. ATP binding to wt, Tr7 and B3 enzymes was similar but this substrate bound only weakly to S1 TK. Each mutant displayed a characteristically distinct pattern of affinities for a range of nucleoside analogue substrates, suggesting that they will show some cross-resistance to drugs which have a similar mechanism of action to acyclovir and BVdU. PMID- 6298345 TI - Can Epstein-Barr virus infect and transform all the B-lymphocytes of human cord blood? AB - Quantitative aspects of Epstein-Barr virus infection and transformation of human neonatal B-lymphocytes have been investigated. 72 to 90% B-cells were obtained with enrichment. Of the B-cells, 19 to 97% showed nuclear antigen (EBNA) 2 days after infection. A difference between different B-cell donors in susceptibility to infection was noted. Analysis of the virus dose-response curves obtained with twofold virus dilutions showed that one virus particle is sufficient to induce EBNA in a cell. Of the infected cells, 50 to 95% multiplied in microtitre wells containing a human fibroblast feeder layer, while only a small proportion established growing colonies in soft agarose, that could be picked up and subcultured. PMID- 6298346 TI - Molecularly cloned bovine papillomavirus DNA transforms mouse fibroblasts in vitro. AB - NIH 3T3 mouse fibroblasts were transformed in vitro by transfection with viral DNA from bovine papillomavirus (BPV) types 4, 2 and 1. The viral DNAs were molecularly cloned in pAT153 to construct pBV4, a recombinant plasmid containing the whole genome of BPV4, pBV2 containing the entire genome of BPV2, and pBV1-D1 which contains the 69% transforming DNA fragments of BPV1. The transformed cells lost contact inhibition, were anchorage-independent, required low serum and were tumourigenic in nude mice. This is the first report of cell transformation in vitro by BPV4 DNA. The recombinant plasmids transformed NIH 3T3 cells with an efficiency of 200 to 300 foci/micrograms DNA. Cleavage of the recombinant plasmids with enzymes that separate the viral DNA from pAT153 DNA did not significantly alter the efficiency of transformation. In all the transformed cells analysed, the recombinant plasmids were found as multiple copies of non integrated monomers. PMID- 6298347 TI - A deletion mutant of mouse mammary tumour virus, lacking 516 nucleotides of the 5' long terminal repeat sequence, can be expressed in a hormone-responsive fashion. AB - In vitro manipulation of proviral DNA of mouse mammary tumour virus (MMTV) was used to construct mutants with defined deletions at the 5' end of the proviral gene. In the mutants 516, 1400 and 2000 nucleotides were removed from the 5' end. The deleted proviral DNA was tested for transcription and glucorticoid hormone regulation of viral RNA expression upon cotransfection into rat XC tk- cells with a thymidine kinase gene. Intact proviral DNA contained in the plasmid vector pBR322 and the deletion mutant pGR16 delta 516, missing 516 nucleotides of the 5' long terminal repeat (LTR) sequence, were transcribed in a hormone responsive fashion and produced RNA species of 35S and 24S. Deletion of the entire LTR sequence abolished MMTV transcription and the hormonal effect. PMID- 6298348 TI - Unusual neutral oligosaccharides in mature Sindbis virus glycoproteins are synthesized from truncated precursor oligosaccharides in Chinese hamster ovary cells. AB - We have previously demonstrated the presence of unusual small asparaginyl oligosaccharides [(Man)3GlcNAc2-ASN] in the mature glycoproteins of Sindbis virus released from both wild-type and lectin-resistant Chinese hamster ovary cells, but the mechanism of synthesis of these structures was not determined. Gel filtration and endo-beta-N-acetylglucosaminidase analyses of Pronase-digested glycopeptides from [3H]mannose-labelled Sindbis virus released at different times after infection of a phytohaemagglutinin-resistant line of Chinese hamster ovary cells demonstrated that these small asparaginyl-oligosaccharides were present in similar relative amounts in virus released throughout the virus infection, rather than arising primarily at late times when cytopathic effects were maximal. Similar analyses of pulse-labelled, cell-associated viral glycopeptides suggested that these small oligosaccharides on mature virus glycoprotein resulted from the normal alpha 1,2-mannosidase processing of truncated precursor oligosaccharides (containing five rather than nine mannoses), rather than from aberrant processing or degradation of the full-size precursor oligosaccharides or normal intermediates. PMID- 6298350 TI - Production of human monoclonal antibody to X31 influenza virus nucleoprotein. AB - In vitro stimulation of human peripheral blood mononuclear cells with X31 influenza virus antigen has been used to enrich for specific anti-X31 antibody producing cells. Following Epstein-Barr virus transformation of these stimulated cells, a cell line which produces human antibody to X31 virus was derived and subsequently cloned. The cloned cells secrete and IgGl kappa antibody which is directed against the nucleoprotein of A type influenza virus. Culture supernatants contain 10 to 20 micrograms/ml of specific antibody which is now used as a standard for the ELISA assay used in our laboratory to detect antibodies to influenza virus. PMID- 6298349 TI - Ontogeny of yellow fever 17D vaccine: RNA oligonucleotide fingerprint and monoclonal antibody analyses of vaccines produced world-wide. AB - Yellow fever 17D vaccines are currently manufactured with approval of the World Health Organization (WHO) in 11 countries. These vaccines have proven highly efficacious and safe. Nevertheless, they have not been fully characterized genetically, a problem for future standardization and modernization of vaccine manufacture now being proposed by WHO. Vaccines in use are derived from two distinct substrains (17D-204 and 17DD) which represent independently maintained passage series from original 17D. In this study, all 17D vaccines produced world wide were characterized by RNA oligonucleotide fingerprinting. Forty-two large oligonucleotides were compared, and differences from an arbitrarily selected reference strain (produced by Connaught Laboratories in the U.S.A.) were determined. With one exception (vaccine produced in South Africa), fingerprints of vaccines derived from substrain 17D-204 were identical. The South African primary seed differed in position of one oligonucleotide, reflecting a charge shift due to a single base change. This difference occurred within one egg passage; a further change in the South African vaccine occurred within one or two passages from primary seed. No antigenic differences between 17D-204-derived vaccines (including South Africa) were demonstrated by neutralization tests using monoclonal antibody. Vaccines derived from the 17DD substrain consistently differed from 17D-204 vaccines in the absence of one oligonucleotide (No. 37). This change probably occurred during 40 additional egg passages in development of the 17DD vaccines. A clear antigenic difference was shown between 17D-204 and 17DD substrain vaccines using monoclonal antibody. 17DD vaccines showed minor genotypic differences, suggesting a higher degree of genetic instability than 17D 204 vaccines. No oligonucleotide fingerprint differences were found between avian leukosis virus (ALV)-free and ALV-contaminated vaccines. No definite genomic correlate of neurovirulence was defined by fingerprinting strains with a history of encephalitic complications in man or of failure to pass monkey neurovirulence tests. Parent Asibi virus showed several oligonucleotide differences and was serologically distinct from 17D vaccine. PMID- 6298351 TI - Analysis of Theiler's virus isolates from persistently infected mouse nervous tissue. AB - The DA strain of Theiler's virus causes a chronic progressive demyelination in mice following intracerebral inoculation. Virus was isolated from chronically infected mice, and then grown in cell culture, and the isolates were compared with the parent virus used for inoculation. No defective interfering particles or temperature-sensitive virus were recovered, and capsid proteins appeared identical by SDS-PAGE. One of three isolates had evidence of genomic mutation by Tl ribonuclease oligonucleotide fingerprinting. The significance of these findings with regard to the generation and maintenance of persistence and to adaptation to cell culture is discussed. Also of interest was the marked difference between the DA fingerprint and that of GD VII, a serologically related strain with different biological activity. PMID- 6298352 TI - Pathogenesis of mouse scrapie: evidence for spread of infection from central to peripheral nervous system. AB - The onset of replication has been studied in spinal cord, dorsal root ganglia and spinal nerves of CW mice infected intraperitoneally with the 139A strain of scrapie. The patterns obtained suggest a centrifugal spread of infection from central to peripheral nervous systems. Infectivity titres in the peripheral nervous system reached a plateau long before the end of the incubation period, and the maximum titres were much lower than in the central nervous system. This suggests that there is a restriction of replication in the peripheral nervous system similar to that already known in extraneural tissues. PMID- 6298353 TI - Analysis of two phosphoproteins related to pp60src from Schmidt-Ruppin D virus particles. AB - During endogenous phosphorylation of partially purified pp60src from virus particles, besides pp60src two additional phosphoproteins, 45K and 42K, were found. These proteins copurify with pp60src. They were shown to be proteolytic degradation products due to the action of the virus-associated protease p15. All three phosphoproteins were present in particles of two different sarcoma virus strains, Schmidt-Ruppin D and Prague C, indicating that this phenomenon is general rather than strain-specific. The degradation rate of pp60src was reduced by the presence of 3 mM-ZnCl2, which acts as a protease inhibitor. PMID- 6298354 TI - Studies on the SV40-like papovavirus SV40-GBM. I. Genomic analysis by restriction endonucleases and electron microscopy after propagation in CV-1 monkey cells. AB - Infection of CV-1 monkey cells with SV40-GBM, a papovavirus isolated from a human glioblastoma multiforme, resulted in the appearance of defective viral DNA molecules. In contrast to SV40 wild-type, two main types of variant DNA molecules could be found after three viral passages at multiplicities of infection of about 10. The molecules of one variant DNA (GBM3-L) were about 19% shorter than the GBM3-H DNA molecules and the DNA of the original GBM isolate, as demonstrated by electron microscopy. Restriction enzyme analysis revealed that GBM3-L DNA had lost both the EcoRI and the HpaII cleavage sites which are located in the late viral genome region. Furthermore, SV40 GBM3-L did not possess the two PvuII sites which are located in the late genome region, and a portion of the GBM3-H and GBM3 L DNA molecules had lost the unique KpnI site. Heteroduplex analysis verified that the rearrangements in the GBM3-L DNA are located only in the late region of this DNA. The possible differences between SV40 wild-type and SV40-GBM are discussed on the basis of these results. PMID- 6298355 TI - Inhibition of vesicular stomatitis virus RNA synthesis by 2',3'-dideoxycytidine 5'-triphosphate. AB - The ability of the compound 2',3'-dideoxycytidine 5'-triphosphate (ddCTP) to serve as an inhibitor of viral RNA synthesis was examined using an in vitro system that supports vesicular stomatitis virus (VSV) protein synthesis, transcription and replication. Viral RNA synthesis was inhibited by 87 and 98% of control, respectively, in reactions containing 1 mM- and 10 mM-ddCTP in place of CTP. VSV RNA replication and transcription were inhibited equally by ddCTP. At a concentration of 1 mM-ddCTP, there was no inhibitory effect on viral protein synthesis; at 10 mM-ddCTP, total protein synthesis was inhibited by 30% as compared to control reactions. The presence of ddCTP had no effect on the size or relative molar amounts of each protein synthesized as analysed by electrophoresis on polyacrylamide gels. This is the first report describing a compound that will inhibit VSV RNA synthesis in vitro without compromising the concurrent synthesis and modification of proteins. PMID- 6298356 TI - Induction of a latent herpes simplex virus from a rat tumour initiated by herpes simplex virus-transformed cells. AB - A rat tumour induced by cells transformed with the sheared DNA of herpes simplex virus (HSV) type 1 HFEM alpha (RE2A) was injected with the intertypic virus HSV-2 HG52 ts 1. Separate plaques, isolated from cocultivation of excised tumour tissue with susceptible cells, yielded virus the DNA of which had the restriction enzyme profile either of the injected HSV-2 virus or that of the HSV-1 virus, originally used to transform the cells. No evidence of in vivo recombination was detected. In Hooded Lister rats, HSV may have the ability to remain in a latent or non replicating state in fibroblasts. PMID- 6298357 TI - Adsorption of purified human cytomegalovirus and induction of early antigens in different cells. AB - Different human and nonhuman cells were assayed for their capacity to absorb human cytomegalovirus (CMV Ad.169) and to support CMV infection in vitro. The CMV adsorptive capacity was assayed by measuring cell-bound radioactivity after addition of purified 3H- or 125I-labeled CMV or by a bioassay for residual infectious virus in the supernatant fluid. Many of the human and nonhuman cell types adsorbed CMV. Induction of CMV early nuclear antigens in the same cells was assayed by anticomplement immunofluorescence staining of fixed cells 1-3 days after infection. CMV early antigens were induced in the human and nonhuman cells that showed a high degree of CMV adsorption. PMID- 6298358 TI - Cold adaptation of parainfluenza virus type 3: induction of three phenotypic markers. AB - In order to attenuate parainfluenza type 3 virus, a wild type strain that was isolated from a child with respiratory disease was adapted to replicate in African green monkey kidney cells at 20 degrees C. Replication at 20 degrees C was not a property of the wild type virus. The virus was serially passaged 45 times in the cold, and clones were selected following passage levels 7, 12, 18, and 45. The population of cold-adapted virus was found to be progressively enriched with temperature sensitive (ts) mutants. After 7 passages in the cold, 1 of 9, and after 12 passages in the cold, 3 of 12 clones were temperature sensitive. Following 18 passages in the cold, 80% of the clones were temperature sensitive and after 45 passages in the cold, all clones were temperature sensitive. In addition to being temperature sensitive each ts clone manifested the tiny plaque morphology. Each temperature-sensitive clone was also cold adapted. Some clones were cold adapted but were not temperature sensitive. The mutants were found to be genetically stable when serially passaged at 32 degrees, 35 degrees, or 39 degrees C. The mutants may possess the necessary degree of attenuation for use as live attenuated intranasal vaccines. PMID- 6298359 TI - Inapparent genital herpes simplex virus infection in college women. AB - During a six-month period, 600 gynecological samples were collected from 585 women with typical herpes lesions, women with non-herpes symptoms (ie, vaginitis, moniliasis, trichomoniasis, etc), and normal women seen at the student health center gynecological clinic and processed for herpes simplex virus (HSV) isolation. From these specimens, 29 samples from 25 of the 585 women (4.3%) were positive for HSV. When these isolates were typed using plaque diameter in chick cells, heat stability of viral thymidine kinase (T.K.), and restriction endonuclease patterns it was found that 18 samples (15 patients or 60%) were HSV 2 and 11 samples (10 patients or 40%) were HSV-1. Inapparent HSV infections constituted 20.0% of the virologically confirmed samples (5 of 25 patients) and represented 0.9% of the total patients studied (5 of 585). The inapparent infections were about equally divided between the two HSV types (2 were HSV-2 and 3 were HSV-1), and 4 of 5 occurred in the presence of clinically diagnosed monilia. PMID- 6298360 TI - Sudden infant death syndrome associated with rotavirus infection. AB - Rotavirus was detected in the stools of five children stricken with sudden infant death syndrome (SIDS) over a three-week period. While none of the children had acute gastroenteritis, four of the five had acute upper respiratory infections. Rotavirus was also identified in tracheal aspirates from two of the infants. Extensive investigations failed to reveal the presence of any other viruses or toxins in specimens obtained from the five children with SIDS. Rotavirus was not found in the stool specimens obtained from a control group of 36 infants including six who died of causes other than SIDS. Future attempts at the prevention of rotavirus infections should be directed at populations susceptible to sudden infant death syndrome. PMID- 6298361 TI - A behavioral program for the management of anorexia and bulimia. AB - The results of a behavioral treatment program for two cases of anorexia and bulimia are presented. The program involved inpatient contingency management for weight gain, thought stopping, flooding, response prevention for bulimia and emesis, and training in self-monitoring skills of caloric intake and weight. The results for both cases indicate that the inpatient program was associated with specific increases in body weight and caloric consumption which were generally maintained at 2-year follow-ups. Flooding and response prevention also appeared to be generally successful in reducing both emesis and bulimia urges and episodes, whereas thought stopping did not appreciably change these behaviors. Serum pituitary gonadotropins (follicle-stimulating hormone and luteinizing hormone) were also restored to normal or near normal levels. Self-monitoring of eating behaviors including caloric intake, emesis, and bulimia seemed to be a useful maintenance strategy and provided timely data for the application of booster treatments during later follow-up intervals. PMID- 6298362 TI - The identification and localization of a catecholamine in the motor neurons of the lobster cardiac ganglion. AB - The cardiac ganglion from Homarus americanus was investigated for the purpose of providing biochemical and histochemical information as to the identity of the neurotransmitter(s) utilized by this system. Three techniques were employed in this study: (1) the glyoxylic acid histofluorescence staining technique (GA), which showed fluorescence characteristic of catecholamines localized in the five motor neurons; (2) high-voltage electrophoresis (HVE) in one dimension followed by ascending chromatography in the second dimension, which indicated incorporation of label from tritiated tyrosine into norepinephrine (NE) and small amounts of dopamine (DA); (3) high-pressure liquid chromatography with electrochemical detection (HPLC/EC), which indicated the presence of endogenous norepinephrine. PMID- 6298363 TI - Calcium-dependent proteolysis in neural tissue is activated at physiologic intracellular Ca2+ levels and inhibited by some anticonvulsants. AB - The level of calcium-activated neutral protease (CANP) activity in the brain and nerve cord of the crayfish Procambarus clarkii was assayed by measuring the degradation of casein yellow by tissue homogenates. When care was taken to maintain the ionic strength of all incubation media at 0.15M and to buffer the Ca2+ activity of the media with 5mM EGTA, CANP was found to be very sensitive to Ca2+; maximal activity was achieved at 1 X 10(-3)M Ca2+, with 50% of this maximum present at the physiologic intracellular Ca2+ activity of 1 X 10(-7)M. We found that the anticonvulsant agent phenytoin was without effect on CANP activity while pentobarbital and a relatively new anticonvulsant agent, valproic acid (an eight carbon branched fatty acid), significantly inhibited CANP activity in a dose dependent manner. The inhibitory effect of valproic acid was shared by a straight chain eight-carbon fatty acid, caprylic acid. These findings demonstrate that inhibition of CANP activity is not limited to agents with a specific molecular structure. They also suggest that CANP plays a role in the normal turnover of proteins that control various cellular functions. PMID- 6298364 TI - Electron micrographic study of precipitates formed by interaction of silicic acid and alkaline phosphatase: contribution to a study of silica urolithiasis in cattle. AB - Association of alkaline phosphatase with silicic acid in precipitates formed in dilute solution was studied as a model for the nonspecific reaction between silicic acid and protein. Precipitates contained 68-83% of the silicic acid and 52-83% of the enzyme in the original mixture and were in the form of aggregates of roundish particles 150-800 nm in diameter. Enzyme protein formed a tightly bound layer on the surface of particles formed in solutions of freshly prepared silicic acid. The similarity between the ultrastructural features of precipitates from solutions of silicic acid and of internal portions of siliceous urinary calculi from cattle suggests that deposition of silica during development of such calculi is due, at least in part, to the interaction of protein with silicic acid in urine. PMID- 6298365 TI - Suriclone: a new cyclopyrrolone derivative recognizing receptors labeled by benzodiazepines in rat hippocampus and cerebellum. AB - Suriclone (RP 31,264), like zopiclone (RP 27,267), belongs to the family of cyclopyrrolones and is chemically entirely different from the benzodiazepines (BZDs). However, it possesses a pharmacological profile close to that of the BZDs and proved to be useful in therapeutics as an anxiolytic agent. In the present paper it is shown that suriclone possesses a high affinity for flunitrazepam binding sites and that tritiated suriclone binds specifically with high affinity in rat hippocampus (KD = 0.44 +/- 0.03 nM) and rat cerebellum (KD = 0.53 +/- 0.12 nM). Further, suriclone binding sites are recognized by BZDs or zopiclone, similarly in the two regions. The affinities of four BZD derivatives--nitrazepam, flunitrazepam, diazepam, and chlordiazepoxide--are similar for suriclone and flunitrazepam binding sites. Suriclone binding sites are, like flunitrazepam sites, protected from thermal inactivation by gamma-aminobutyric acid (GABA) (10 microM), but only flunitrazepam binding is enhanced by GABA. It could be postulated from this that suriclone interacts with a subpopulation of receptors that might be modulated differently from flunitrazepam binding sites. Our results indicate that suriclone could be a new probe for investigating the so-called BZD receptors. PMID- 6298366 TI - Formation of a persistent inhibitory state of brain adenylate cyclase by GTP analogs. AB - Addition of 10 microM guanyl-5'-ylimidodiphosphate at 30 degrees or 0 degree to guinea pig brain particulates instantaneously evoked nearly 50% inhibition of adenylate cyclase activity as determined after removal of the GTP analog by washing of the particulates. The inhibitory state, once formed, persisted for at least 60 min as long as the preparation was kept either in a medium devoid of the analog (0-30 degrees) or in its presence at 0 degree. During incubation at 30 degrees in the presence of the analog, however, the inhibited or nontreated enzyme showed a gradual increase in enzyme activity. Both the inhibitory and the activating effects of the analog were saturable, with a half-maximal concentration of about 1.0 microM, and were antagonized by simultaneous addition of GTP, GDP, and GMP (in decreasing order). The persistently inhibited enzyme enabled the detection of marked stimulation by norepinephrine and histamine, whereas these amines showed only marginal stimulation of the enzyme before treatment with the analog. Formation of such a persistent inhibitory state appears to be specific to brain cyclase. PMID- 6298367 TI - Effect of some calcium antagonists on muscarinic receptor-mediated cyclic GMP formation. AB - Several calcium antagonists were screened for their effect on muscarinic acetylcholine receptor-mediated cyclic GMP formation in murine neuroblastoma cells (clone N1E-115). Mn2+, Ni2+, and verapamil rapidly antagonized the response noncompetitively, with the following order of potency: verapamil greater than Mn2+ greater than Ni2+. The effects of Mn2+ and Ni2+, but not those of verapamil, were largely reversed by increasing extracellular calcium concentration. Additional effects of these agents included displacement of [3H]quinuclidinyl benzilate binding by verapamil and elevation of cyclic GMP levels by Mn2+ and Ni2+ in the absence of agonists. These results are in support of the hypothesis that receptor-mediated cyclic GMP formation by these cells is dependent upon entry of calcium into the cell and demonstrate the complexity of the effects of calcium antagonists. PMID- 6298368 TI - Phosphorylation of brain synaptic and coated vesicle proteins by endogenous Ca2+/calmodulin- and cAMP-dependent protein kinases. AB - Phosphorylation of brain synaptic and coated vesicle proteins was stimulated by Ca2+ and calmodulin. As determined by 5-15% sodium dodecylsulfate (SDS) polyacrylamide gel electrophoresis (PAGE), molecular weights (Mr) of the major phosphorylated proteins were 55,000 and 53,000 in synaptic vesicles and 175,000 and 55,000 in coated vesicles. In synaptic vesicles, phosphorylation was inhibited by affinity-purified antibodies raised against a 30,000 Mr protein doublet endogenous to synaptic and coated vesicles. When this doublet, along with clathrin, was extracted from coated vesicles, phosphorylation did not take place, implying that the protein doublet may be closely associated with Ca2+/calmodulin dependent protein kinase. Affinity-purified antibodies, raised against clathrin used as a control antibody, failed to inhibit Ca2+/calmodulin-dependent phosphorylation in either synaptic or coated vesicles. Immunoelectron cytochemistry revealed that this protein doublet was present in axon terminal synaptic and coated vesicles. Synaptic vesicles also displayed cAMP-dependent kinase activity; coated vesicles did not. The molecular weights of phosphorylated synaptic vesicle proteins in the presence of Mg2+ and cAMP were: 175,000, 100,000, 80,000, 57,000, 55,000, 53,000, 40,000, and 30,000. Based on the different phosphorylation patterns observed in synaptic and coated vesicles, we propose that brain vesicle protein kinase activities may be involved in the regulation of exocytosis and in retrieval of synaptic membrane in presynaptic axon terminals. PMID- 6298369 TI - An investigation of the antiviral activity of Podophyllum peltatum. AB - The antiviral activity of an aqueous extract of Podophyllum peltatum was investigated. The extract was fractionated by reversed-phase chromatography, and podophyllotoxin was found to be the most active component in inhibiting the replication of measles and herpes simplex type I viruses. beta-Peltatin and desoxypodophyllotoxin produced marginal antiviral effects, while alpha-peltatin and picropodophyllotoxin were inactive at the levels tested. PMID- 6298370 TI - Virus antibodies in the cerebrospinal fluid of multiple sclerosis patients detected with ELISA tests. AB - The enzyme-linked immunosorbent assay (ELISA) was used to determine levels of specific IgG antibodies against measles, rubella, vaccinia, corona (OC43) and mumps viruses in cerebrospinal fluid (CSF) and serum of 18 patients with clinically definite multiple sclerosis (MS), 8 patients with optic neuritis (ON), 27 patients with other neurological disease (OND), and 88 control subjects without central nervous system disease. Serum antibody levels were not significantly different between the four groups. Differences in the frequency and levels of CSF antibodies between the four groups were observed. Control patients had serum/CSF antibody ratios from 2.0 to 3.0 (log) with an average of 2.5 corresponding to a 320-fold difference between serum and CSF antibody levels. MS patients had ratios from 1.1 to 2.1 with an average of 1.6. The average was 2.0 for the ON patients. The average for the OND patients was similar to the controls. The altered serum/CSF ratios for several viruses within an individual patient was similar. These results suggest that nonspecific immunostimulation is responsible for the increased levels of CSF virus antibodies. PMID- 6298371 TI - Blood brain barrier disturbance and immunoglobulin G levels in the cerebrospinal fluid of the mouse following peripheral infection with the demyelinating strain of Semliki Forest virus. AB - Immunoglobulin G and albumin levels have been measured in the cerebrospinal fluid and serum of Swiss A2G mice following single intraperitoneal inoculation of Semliki Forest virus. This strain of virus used causes a meningoencephalitis followed by immunologically-mediated demyelination. By the use of the levels of immunoglobulin G and albumin in cerebrospinal fluid and serum to calculate the cerebrospinal fluid: serum ratios and the cerebrospinal fluid immunoglobulin G index, it has been shown that the blood-brain barrier breakdown is mild and restricted to within the first 7-8 days after virus inoculation when the inflammatory response is maximal. Immunoglobulin G index provides a measure of synthesis of immunoglobulin G within the blood-brain barrier. Synthesis has been shown to occur from day 10 onwards up to at least day 61 following infection. Arboviruses are known to persist in central nervous system tissue and it is suggested that a continuing immune response within the central nervous system occurs and is perpetuated by persisting virus. Because of the mode of replication of Semliki Forest virus it is also possible that some of the immune response is directed against central nervous system components. PMID- 6298372 TI - Time course of improved neuromuscular function following plasma exchange alone and plasma exchange with prednisone/azathioprine in myasthenia gravis. AB - Ten patients with myasthenia gravis (MG) were treated with weekly plasma exchange (PE) in combination with prednisone and azathioprine; 4 of the patients were treated with 3-6 PE alone, before instituting prednisone/azathioprine. Four clinico-physiological parameters of neuromuscular function as well as the anti acetylcholine receptor antibody (anti-AChRAb) titers were measured 1-3 times per week. Evidence of improved neuromuscular function (NMF) was based on: (1) reduced % decrement of the maximum muscle compound potential following 2/s supramaximal nerve stimulation; (2) increased maximum force; (3) increased outstretched arm time, and (4) increased vital capacity. An early improvement in NMF occurred within the first 2 weeks (sometimes as early as 1-2 days), which was quantitatively comparable in patients treated with PE alone and PE plus prednisone/azathioprine. This early improvement reached a plateau, and was subsequently maintained for 3-4 weeks, followed by a later substantial improvement after 8-12 weeks of combined PE plus prednisone/azathioprine. PMID- 6298373 TI - Response properties of thin myelinated (A-delta) fibers in human skin nerves. PMID- 6298374 TI - Effect of norepinephrine depletion of hippocampus on neuronal transmission from perforant pathway through dentate gyrus. PMID- 6298376 TI - Composite EPSPs in motoneurons of different sizes before and during PTP: implications for transmission failure and its relief in Ia projections. PMID- 6298375 TI - Event-related extracellular potassium ion activity changes in frontal cortex of the conscious cat. PMID- 6298377 TI - Sensitization of unmyelinated nociceptive afferents in monkey varies with skin type. PMID- 6298378 TI - Ca++/calmodulin-regulated protein phosphorylation in the Aplysia nervous system. AB - Protein substrates for an endogenous CA++/calmodulin-dependent protein kinase were characterized in the Aplysia nervous system. Ethylene glycol bis(beta aminoethyl ether)-N,N,N',N'-tetraacetic acid-washed membrane fractions from Aplysia ganglia contain an endogenous Ca++/calmodulin-dependent protein kinase which phosphorylates a number of membrane proteins. Such washed membrane preparations exhibit little or no adenosine 3':5'-cyclic phosphate (cAMP) dependent protein kinase activity but do contain substrates for exogenously added catalytic subunit of cAMP-dependent protein kinase. Low concentrations of catalytic subunit rapidly stimulate the phosphorylation of a protein of Mr 70,000 and of a 52,000-dalton doublet, indicating that these proteins are major substrates for this enzyme. Phosphopeptide patterns obtained after limited proteolysis suggest that the 70,000-dalton protein and the 52,000-dalton doublet are similar in structure and that their phosphorylation is stimulated both by Ca++/calmodulin and by catalytic subunit. The 52,000-dalton doublet consists of two closely spaced bands: the phosphorylation of the upper band is stimulated by catalytic subunit, whereas the phosphorylation of the lower band is stimulated by the endogenous Ca++/calmodulin-dependent protein kinase. The results suggest that in Aplysia membranes a number of proteins can be phosphorylated by both Ca++/calmodulin-dependent protein kinase and catalytic subunit of cAMP-dependent protein kinase. This convergence of biochemical effects of Ca++ and cAMP may play a role in some of their physiological actions in molluscan neurons. PMID- 6298380 TI - The pterygo-maxillary region (fossa infra-temporalis). A clinical, radioanatomical and pathological study. PMID- 6298379 TI - Chemical characteristics of the L-glutamate receptor on the Onchidium neuron. AB - Application of L-glutamate (L-glu) to particular neurons (G-H cells) in the esophageal ganglia of Onchidium produced hyperpolarization associated with an increase in K+ permeability. The reversal potential (Erev) for this response, was -60 mV. The ED50, an indicator of affinity between L-glu and its receptive site, was 5.3 mM, and the Hill coefficient n, an indicator of cooperativity, was 2. Other chemicals, structurally analogous to L-glu, were also examined on G-H cells. The responses of L-aspartate were similar to those of L-glu with respect to Erev, ED50, and n. However, the G-H cells responded differently to D-glutamate (D-glu) and kainic acid. The D-glu response had an Erev = -37.5 mV, ED50 = 29.4 mM, and n = 1; that of kainic acid had an Erev = 0 mV, ED50 = 1.5 mM, and n = 1. L-glutamine and GABA elicited very small responses, amplitudes of which were less than 10% of the L-glu response. The structure-activity relationship between L-glu and its analogues, estimated from Erev, ED50, and Hill coefficient during each drug application, indicates that the presence of alpha-NH2, alpha-COOH, and gamma COOH groups is essential for the L-glu response and that the response characteristics may be closely related to the relative positions of these groups. PMID- 6298381 TI - Inhibition by dipyridamole of cerebral vasospasm induced in vitro by whole blood. AB - This study evaluates the effect of dipyridamole, an inhibitor of platelet aggregation, on cerebral artery contraction induced in vitro by the addition of whole blood. Whole fresh arterial blood added to isolated rabbit basilar artery bathed in a physiological buffer produces a sustained contraction in vessels observed for 60 minutes. Significant dose-dependent inhibition of contraction was observed when dipyridamole was added to the vessel bath. This effect was not influenced by preincubating cerebral vessels with aspirin, an inhibitor of prostaglandin synthesis. It is suggested that inhibition of whole blood-induced cerebral artery contraction by dipyridamole does not result from potentiation of cerebral vessel prostaglandin pathways, but possibly from a direct effect on platelets. PMID- 6298382 TI - Primary cerebellar glioblastoma multiforme with an unusually long survival. Case report. AB - Light microscopic and electron microscopic findings of a primary cerebellar glioblastoma multiforme (GM) are presented. An infiltrating tumor of the vermis and the right cerebellar hemisphere had been partially removed from a 39-year-old woman. Postoperative irradiation to the posterior fossa enabled her to work for the next 5 1/2 years. At readmission, progressive pontocerebellar signs were observed. In spite of repeated irradiation and intrathecal chemotherapy, she died after 1 month. Autopsy revealed extensive tumorous infiltration of the right cerebellar hemisphere, pons, and medulla. Both the biopsy and autopsy specimens showed typical features of GM. Tumorous propagation resulted in extreme enlargement of the right inferior olive. Electron microscopic analysis disclosed characteristic bundles of glial filaments, cytoplasmic inclusions lying within nuclear folds, and intracytoplasmic granules of uncertain nature. The possible cause of the long survival is discussed and a comparison is made with previously reported cases. PMID- 6298383 TI - Meningeal malignant fibrous histiocytoma arising from a thoracolumbar myelomeningocele. Case report. AB - The authors present a case of malignant fibrous histiocytoma (MFH) involving a thoracolumbar myelomeningocele. The patient died of metastases 11 months after diagnosis, in spite of extensive surgery, radiation therapy, and chemotherapy. Meningeal involvement by MFH is very rare; 11 additional cases are reviewed. Glial fibrillary acidic protein staining is critical for proper histological diagnosis, as this tumor can be easily confused with pleomorphic xanthoastrocytoma or glioblastoma involving the meninges. Malignant degeneration within a myelomeningocele is also very rare. Eight additional cases from the literature are reviewed. Our case is the only one with MFH. Chronic irritation to the unrepaired congenital defect appears to be a key to the development of the malignancy. Prophylactic repair in infancy might prevent the occurrence of malignant degeneration. PMID- 6298384 TI - Fourier amplitude ratio: a new way to assess valvular regurgitation. AB - The stroke-volume ratio determined from the equilibrium gated blood-pool study has been utilized to assess valvular regurgitation, but it is difficult to get reproducible results using generally available equipment. We have developed a new approach utilizing the Fourier amplitude ratio of the left and right ventricles, which is easily implemented and reproducible. Initial clinical experience shows that 17 patients with valvular regurgitation were clearly distinguished from 30 patients without valve disease. PMID- 6298385 TI - Radionuclide angiographic demonstration of systemic lung arterialization with arteriovenous fistulas. AB - Radionuclide angiography has diagnosed systemic arterialization of the right lung base in a patient presenting with a basal thoracic bruit. The bruit was due to high flow in the systemic artery and development of arteriovenous fistulas, confirmed by TCT scan and contrast aortography. But the parenchyma of the right lung base appeared normally aerated on the radiographic studies, and Xe-133 ventilation scintigraphy was normal. This case was therefore classified as systemic arterialization of lung without sequestration. PMID- 6298386 TI - Influence of diet or intrarectal bile acid injections on colon epithelial cell proliferation in rats previously injected with 1,2-dimethylhydrazine. AB - The effects of varying colon bile acid concentrations on rat colon epithelial cell proliferation were studied. Bile acid concentrations were altered by intrarectally injecting either deoxycholic or lithocholic acid for 4 weeks or by increasing the dietary fat or fiber (wheat bran, agar, or carrageenan) intake for 4 weeks. 1,2-Dimethylhydrazine (DMH) was s.c. injected into half of the rats 1 week before treatments began. Colon epithelial cell proliferation was measured by [3H]thymidine autoradiography of colon crypts. Rats injected with DMH had more DNA-synthesizing cells per crypt. Neither bile acid injection nor any of the diets altered the number of DNA-synthesizing cells per crypt. DMH injections, deoxycholic and lithocholic acid intrarectal injections, and dietary agar and wheat bran all increased the total number of cells per crypt. High fat diets and dietary carrageenan did not affect cell number. All diets containing fiber lowered total fecal bile acid concentrations, but increasing the fat content of the diet did not affect them. These results indicate that the bile acid injections and dietary agar and wheat bran induce a slight hyperplasia in the colon. PMID- 6298387 TI - The involvement of liver fructokinase in the metabolism of D-xylulose and xylitol in isolated rat hepatocytes. AB - Hepatocytes isolated from fed, male, Sprague-Dawley rats accumulate xylulose-1 phosphate and glycolaldehyde as well as xylulose-5-phosphate when incubated with 2-20 mM D-xylulose. Fructokinase inhibitors (fructose and 1-deoxyfructose) decreased xylulose-1-phosphate and glycolaldehyde (but not xylulose-5-phosphate) levels in xylulose-treated hepatocytes, demonstrating the role of fructokinase in xylulose-1-phosphate and glycolaldehyde formation. As the fructokinase inhibitors had no overall effects on the conversion of D-xylulose to glucose, the overall flux through the pathway involving fructokinase was less than 27% of the total D xylulose utilized. In hepatocytes from fed or fasted rats there was no detectable accumulation of either xylulose-1-phosphate or glycolaldehyde after treatment with 20 mM xylitol. Other differences between xylitol and D-xylulose metabolism in rat hepatocytes included a slower rate of xylitol metabolism in all preparations and a difference in the relative conversion of xylitol to glucose in hepatocytes from fasted rats. Rats adapted to 20% xylitol (diarrhea-free) had a lower water consumption than those fed a control cornstarch diet; there were no differences in weight gain, food consumption or in rates or metabolite patterns of xylitol metabolism in hepatocytes isolated from these rats. Despite the minor role of fructokinase in the overall metabolism of xylitol and of D-xylulose as shown by these results, it is not possible to exclude the possibility of some flux through the pathway involving xylulose-1-phosphate and glycolaldehyde formation as a possible route for oxalate formation. PMID- 6298388 TI - Growth performance and intestinal transit time of rats fed purified and natural dietary fibers. AB - The effects of some selected purified fibers were compared to those derived from cereals or legume seeds. Rats were fed for at least 9 weeks and measurements were taken to determine feed consumption, weight gain, feed efficiency ratios (FER), protein efficiency ratios (PER), apparent protein digestibility, and rate of transit through the gastrointestinal tract. Most diets were designed to contain approximately 10% dietary fiber and 10% protein. Compared to the fiber-free diet, pectin reduced weight gain, FER, PER and apparent protein digestibility values. Cellulose, xylan and raffinose had no influence on feed intake, weight gains or FERs. However, cellulose and xylan increased PER values and the rates of food passage but decreased the apparent protein digestibility values. Feed utilization, protein digestibility and growth were similar for the wheat bran, corn bran and fiber-free diets. These cereal fibers caused the rates of transit to be significantly increased relative to the fiber-free control diet. The hull and cell-wall-fiber fractions of beans, when compared to the fiber-free diet, had little effect on feed consumption, growth, FER or PER. The cell-wall-fiber fraction reduced apparent protein digestibility and the hull fraction accelerated food passage relative to the fiber-free diet. PMID- 6298389 TI - Effects of dietary fibers on mineral status of chicks. AB - We investigated the effects of replacing dietary starch with dietary fibers of various types and particle sizes on utilization of bivalent cations by chicks. Inclusion of polyethylene powder, alfalfa cell walls, corn bran, oat bran and wheat bran into diets resulted in lower serum and tibia Zn levels. However, addition of pectin, cellulose and a xylose:gum arabic (50:50) mixture did not affect tissue Zn levels, whereas addition of 4% lignin increased serum Zn level from 2.50 to 3.44 micrograms/ml. Ingestion of fibers with particle sizes less than 300 micrometers resulted in higher serum Zn levels compared with those of particle sizes greater than 300 micrometers. Incorporation of 4 and 8% pectin decreased (P less than 0.05) serum and tibia Mg levels. When brans were fed, kidney Mg levels were lower due to fiber additions. Inclusion of lignin, the xylose:gum arabic mixture and alfalfa cell walls resulted in lower liver Cu levels; polyethylene and solka floc had no effect on liver Cu status. Fiber did not affect tissue Ca or Fe status. Each mineral influenced by fiber type, level or particle size appeared to be affected through different mechanisms. PMID- 6298390 TI - Glycerol kinase activity in adipose tissue of obese rats and mice: effects of diet composition. PMID- 6298391 TI - Mechanisms of toxicities of some detergents added to a diet and of the ameliorating effect of dietary fiber in the rat. AB - Mechanisms of the adverse effects of dietary Tween 20, Tween 60, Span 20, sodium taurocholate (NaTC), sodium deoxycholate (DOC), sodium laurylbenzene sulfonate (LBS) and sodium dodecyl sulfate (SDS), and of the ameliorating effect of the concurrent feeding of dietary fiber were investigated along with releases of the hydrolase activities, which were localized in the brush border membrane of rat small intestine, during a jejunum perfusion in vivo. The releases of sucrase, maltase and alkaline phosphatase activities from the jejunum with Ringer bicarbonate solution (RBS) perfusion for 150 min proceeded at a constant rate after RBS perfusion for the first 30 min. The detergents were perfused after RBS perfusion for 60 min. In Tween 20- or 60-RBS perfusion at the 2% level, the released sucrase activity gradually increased, reaching a level 3 times that with RBS perfusion 90 min after the beginning of Tween 20- or Tween 60-RBS perfusion. With NaTC- or DOC-RBS perfusion at the 0.5% or 0.2% level respectively, the released sucrase activity reached a level 3 to 4 times that with RBS perfusion within 30 min of the beginning of NaTC- or DOC-RBS perfusion, but that with Span 20-RBS perfusion at the 2% level was slightly lower compared with that with RBS perfusion. On the other hand, with SDS- or LBS-RBS perfusion at the 0.5% level, the released alkaline phosphatase activity rapidly reached a level 3 to 4 times as high as that with RBS perfusion. The inclusion of Gobo dietary fiber at the 0.04% level with Tween 20-RBS perfusion completely eliminated the releasing effect of Tween 20 on sucrase activity. These results suggest that the primary cause of the adverse effects of the feeding of these detergents is the exfoliating or releasing effect thereof on the brush border membrane, together with the inhibitory effect of some of these detergents on intestinal disaccharidase activities, and that the dietary fiber prevents the exfoliating or releasing effects of several detergents on the brush border membrane. PMID- 6298392 TI - Myeloid bodies in drug-induced acute tubular necrosis. AB - A growing list of drugs, metals, and chemicals has been implicated as the cause of functional and structural damage specifically to the proximal tubular epithelium. Renal biopsies were obtained from three patients who had developed nephrotoxic agent-related acute renal failure. Two of the patients had received gentamicin and viomycin; the third patient had heavy exposure to chromium. All three biopsies showed acute tubular necrosis (ATN) on light microscopy. Electron microscopy revealed that the proximal tubular cells and, to a lesser degree, the distal tubular cells, contained abundant, variably sized myeloid bodies. In our previous experimental study of viomycin-induced ATN in rats, similar ultrastructural findings of a gradual increase in the number of myeloid bodies in the proximal tubular cells were also observed. The constant presence of myeloid bodies in the tubular epithelial cells following drug-induced tubular necrosis suggests that they may represent lysosomal isolation of drug-bound cytoplasmic structures, as a cellular mechanism to degrade toxic substances and, therefore, may serve as an ultrastructural marker of cellular drug uptake and drug disposition. PMID- 6298393 TI - Lysozyme and angiotensin converting enzyme levels in experimental mycobacterial granulomas. AB - A study has been made on mycobacterial-induced granulomas in guinea-pig lymph nodes. Lysozyme and angiotensin-converting enzyme (ACE) were measured in the auricular lymph nodes and serum of guinea-pigs which had received live BCG (Pasteur) or Cobalt (Co)-irradiated armadillo-derived Mycobacterium leprae intradermally into the ear or dinitrofluorobenzene (DNFB) painted epicutaneously upon the ears. In the lymph nodes with granulomas induced by either live BCG or killed M. leprae, the mean concentrations of lysozyme and ACE varied directly with the mean weight of the lymph nodes but the temporal pattern of weight change differed with the two agents. In M. leprae recipients at the time of peak lymph node weight, serum lysozyme and ACE values were significantly greater than those observed in controls; in animals receiving live BCG (Pasteur), serum lysozyme but not ACE values were elevated significantly at the time of peak lymph node weight. Four days following the epicutaneous application of DNFB, where there was no granuloma, there was a similar increase in the concentration of lysozyme and ACE in the lymph nodes. At the same time, there was also significant elevation in the serum lysozyme and ACE concentrations. Thus, in the granulomatous responses, the parallel tissue and serum changes in lysozyme and ACE concentrations were consistent with increased production and secretion of each enzyme by cells of the mononuclear phagocyte series. The increased lysozyme and ACE concentrations found in the lymph nodes of DNFB sensitised animals gives further evidence that such changes are not unique to granulomas. Finally, the intradermal administration of dead M. leprae in guinea pigs also produced increased lysozyme and ACE levels similar to that found in leprosy in man. PMID- 6298394 TI - Pathological aspects of cholangiocarcinoma. AB - Tissue specimens resected from liver and bile-ducts were examined in an attempt to distinguish features of cholangiocarcinoma from those of other tumours and from reactions to duct obstruction and changes in the tumour-bearing liver. Macroscopical and microscopical features were compared and certain changes were found only in cases of cholangiocarcinoma. The type of tumour correlated moderately well with the level of bile duct involved and some microscopical findings were noted only in patients suffering from cholangiocarcinoma. Mucin was demonstrable in all 33 cholangiocarcinomas. Other striking features included heterogeneity of epithelial cells within the same acinus and a tendency to spread between hepatocyte plates, along duct walls and in relation to nerves. Reactive hyperplasia of ductal subepithelial mucous glands was found in association with duct obstruction with and without cholangiocarcinoma. Sclerosis, a common accompaniment of cholangiocarcinoma was noted in other tumours. PMID- 6298395 TI - Immunohistochemical localization of adenosine 3':5'-cyclic monophosphate in female ixodid tick Amblyomma americanum (L.) salivary glands. AB - Localization of adenosine 3':5'-cyclic monophosphate (cyclic AMP) in alveoli of salivary glands of female Amblyomma americanum (L.) was accomplished with an indirect immunofluorescent technique. Little cyclic AMP fluorescence was seen in Type I alveoli in glands of unfed females but considerable fluorescence was seen in Type I alveoli of glands obtained from females that had fed. The most intense cyclic AMP fluorescence was observed in complex granular cells of Type II and III alveoli in glands of unfed females and glands from females in early stages of tick feeding. In the latter stages of tick feeding an increase in fluorescence in Type III alveoli was observed in cells near the lumen, possibly adluminal interstitial or transformed granular cells. PMID- 6298396 TI - Vasoactive intestinal peptide producing neuroblastoma. AB - The Verner-Morrison syndrome has been described in 19 previous patients with ganglioneuroma and ganglioneuroblastoma but never neuroblastoma. Its occurrence following treatment of a neuroblastoma with chemotherapy with maturation of the tumor has only been reported on one previous occasion. Our case suggests that vasoactive intestinal polypeptide may be used not only as a diagnostic indicator for the presence of a neural crest tumor but also as a marker to monitor maturation of the tumor and indicate an improving prognosis. PMID- 6298397 TI - Therapy and outcome in 51 children with mesoblastic nephroma: a report of the National Wilms' Tumor Study. AB - Mesoblastic nephroma is a distinctive pathologic renal tumor with unique clinical, therapeutic, and prognostic patterns. This report reviews 51 patients (2.8% of 1905 patients submitted to NWTS with renal tumors) with this diagnosis from the NWTS contrasting this disease entity with Wilms' tumor. There were 33 males and 18 females, predominantly term babies born after uncomplicated pregnancies. The mean age at tumor excision was 3.44 +/- 0.6 mo (one child 9 yr). A palpable mass was the predominant presentation in 48 patients and in addition hematuria (9), hypertension (2), vomiting (3), and jaundice (1) were noteworthy. Diagnostic studies included IVP (49 positive, 2 negative) and ultrasound (15 positive, 1 negative). Adequate operative excision was achieved in 43 of 51 patients while 8 children had local extension and 10 had tumor spillage. Operation alone (23), predominantly since 1978, surgery plus chemotherapy (24), prior to 1978, and surgery, chemotherapy, and radiation therapy (4), prior to 1976, were the modes of therapy. The follow-up ranges from 4 mo to 11.5 yr. Survival was excellent; 50 patients survive (98%), only 1 dying of sepsis. These data suggest that mesoblastic nephroma contrasts with Wilms' tumor in that it occurs in a younger age group, has a benign biologic behavior, and a more favorable outcome. Aggressive multimodal treatment though utilized in the early years of the study is not essential to achieve this outcome. PMID- 6298398 TI - Stage III Wilms' tumor of a solitary kidney in a patient with Marfan's syndrome: a 5-yr survival. PMID- 6298399 TI - Effects of p-chloromercuribenzene sulfonate on the uptake of D- and L-leucines in Ehrlich ascites tumor cells. AB - In order to ascertain the significance of the hydrophobic region in the carrier protein responsible for the D-leucine transport reported previously, the effects of p-chloromercuribenzene sulfonate (PCMBS) were investigated on the transport of D- and L-leucines in Ehrlich ascites tumor cells. The uptake of D-leucine in the presence of PCMBS was inhibited more strongly than that of L-leucine. Km values for D-leucine uptake increased with increasing concentration of PCMBS, while those for L-leucine showed only a little increase. D-Leucine uptake was restored considerably from the PCMBS inhibition by washing with buffer, whereas L-leucine uptake was only slightly affected. The inhibition of D-leucine exit by PCMBS was also higher than that of L-leucine. These results suggest that the binding site of PCMBS for the carrier protein of Ehrlich cells is involved in its hydrophobic region that would be more significant for the binding of alkyl side chain D leucine than that of L-leucine. PMID- 6298400 TI - Kinetics and mechanism of degradation of cefotaxime sodium in aqueous solution. AB - The degradation kinetics and mechanism of a potent new cephalosporin, cefotaxime sodium, in aqueous solution were investigated at pH 0-10 at 25 degrees and an ionic strength of 0.5. The degradation rates were determined by high-pressure liquid chromatography and were observed to follow pseudo first-order kinetics with respect to cefotaxime sodium concentration. The data suggested that the rate of degradation was influenced significantly by solvolytic, hydrogen ion, and hydroxide ion catalysis. No primary salt effects were observed in the acid or neutral regions; however, a positive salt effect was observed at pH 8.94. Buffer catalysis due to the buffer species employed was not seen during the kinetic studies. The pH-rate profile at 25 degrees indicated that the maximum stability of cefotaxime sodium occurred in the pH 4.5-6.5 region. In aqueous solution, cefotaxime was shown to degrade by two parallel reactions: de-esterification at the C-3 position and beta-lactam cleavage. Good agreement between the theoretical pH-rate profile and the experimental data support the proposed degradation process. PMID- 6298401 TI - Synthesis and biological activity of an amino analogue of a tripeptide inhibitor of angiotensin-converting enzyme. AB - An amino analogue of N-benzoyl-phenylalanyl-glycyl-proline, a tripeptide inhibitor of angiotensin-converting enzyme, was synthesized. The analogue (III) has the phenylalanyl-glycine amide linkage of N-benzoyl-phenylalanyl-glycyl proline reduced to a methylene amine. Compound III was tested as an inhibitor of porcine plasma angiotensin-converting enzyme and has an I50 of 620 microM compared with an I50 of 9.6 microM for its parent tripeptide. These results are explained in terms of a proposed model of the converting-enzyme active site. PMID- 6298402 TI - Catecholamine-induced renin release in the anesthetized mongrel dog is due to both alpha and beta adrenoceptor stimulation: evidence that only the alpha adrenoceptor component is prostaglandin mediated. AB - The role of renal alpha and beta adrenoceptor activation and prostaglandin synthesis in mediating renin release to intrarenal infusions of the natural neurotransmitters, norepinephrine and epinephrine, was assessed in anesthetized mongrel dogs. Intrarenal infusions of norepinephrine and epinephrine at doses adjusted to reduce renal blood flow by 20 and 50% of baseline values elicited renin release that was not completely blocked by either alpha adrenoceptor blockade with phentolamine or beta adrenoceptor blockade with propranolol. The renin release that persisted during propranolol administration was abolished by the alpha adrenoceptor antagonist, phentolamine, and by the prostaglandin synthetase inhibitor, indomethacin. The beta adrenergic component of renin release, elicited in the presence of phentolamine, was not blocked by indomethacin but abolished by propranolol. These data are consistent with the hypothesis that norepinephrine and epinephrine stimulate renin release by activation of both the renal beta and renal alpha adrenoceptors. The beta adrenoceptor-stimulated renin release appears to be direct and independent of the prostaglandin system, whereas the alpha adrenoceptor-stimulated renin release appears to be indirect and dependent on the generation of endogenous prostaglandins. PMID- 6298403 TI - Qualitative and quantitative differences between the postsynaptic alpha adrenoceptors of rabbit ear artery and thoracic aorta. AB - The alpha adrenoceptor properties of the rabbit ear artery and thoracic aorta were assessed using isolated blood vessel rings mounted in tissue baths. Labetalol, an alpha and beta receptor antagonist, caused dose-dependent contractions in control, reserpinized and surgically denervated ear arteries. This contraction was inhibited by phentolamine and abolished by the irreversible alpha receptor antagonist, N,N'-bis-(O-methoxy-benzylaminohexyl) cystamine tetrahydrochloride. Thoracic aorta failed to respond to labetalol. Using labetalol as an antagonist against methoxamine, labetalol pA2 values were 7.4 +/- 0.3 (95% confidence interval) and 7.13 +/- 0.25 in ear artery and aorta, respectively. Thus, labetalol had the same affinity for the alpha receptors of these two vessels but was an alpha agonist only in the ear artery. Norepinephrine ED50 values and dissociation constants (KA) were determined by analysis of dose response data with and without partial inactivation of alpha receptors by phenoxybenzamine. Ear artery and aorta norepinephrine ED50 values, 4.24 (2.24 8.03) X 10(-8) M and 2.48(1.64-3.76) X 10(-8) M, respectively, were not significantly different. In contrast, norepinephrine KA values differed by a factor of 32, 4.11 (3.02-5.60) X 10(-6) M and 1.29 (0.85-1.94) X 10(-7) M, respectively. Receptor reserves were also markedly different in these vessels. Thus, ED50 was achieved with 1% receptor occupancy in ear artery as compared to 16% receptor occupancy in aorta. It is concluded that the alpha receptors of ear artery and aorta are both qualitatively and quantitatively different. PMID- 6298404 TI - Effect of aminophylline on amphotericin B nephrotoxicity in the dog. AB - The polyene antibiotic, amphotericin B, causes an acute reduction in renal blood flow and glomerular filtration rate. The purpose of this study was to evaluate the hypothesis that the renal vascular response to amphotericin B can be blocked by aminophylline. Toward this end, the effect of aminophylline on the renal response to amphotericin B in sodium-depleted dogs was examined. In dogs not treated with aminophylline, amphotericin B (0.5 mg/kg infused i.v. over 20 min) significantly reduced renal blood flow and glomerular filtration by 49.9 +/- 12.6 ml/min (mean +/- S.E.M.) and 23.4 +/- 2.4 ml/min, respectively at 140 min after the amphotericin B infusion. In dogs treated with an intrarenal aminophylline infusion (5 mg/min), the renal blood flow and glomerular filtration rate response to amphotericin B did not differ from that of amphotericin B vehicle. We conclude that aminophylline inhibits the renal response to amphotericin B. The possible clinical relevance of these observations are discussed. PMID- 6298405 TI - Metabolism of [14C]paraldehyde in mice in vivo, generation and trapping of acetaldehyde. AB - The metabolic fate and the kinetics of paraldehyde metabolism after the i.p. administration of a 400 mg/kg dose of this agent were investigated in mice. Paraldehyde was found to have a biologic half-life in this species of 41.5 min, its disappearance from blood being governed by a single component exponential process with a rate constant of 0.0167 min-1. By using [14C]paraldehyde, it was found that the major process responsible for paraldehyde disappearance was its metabolic degradation to carbon dioxide, a two-step process; the first step of which was inhibited by pretreatment with SKF-525A. The rate constants for the two steps being 0.0121 and 0.0212 min-1, respectively; on the basis of these rate constants it was calculated this pathway would account for 72.3% of the administered dose at infinite time. A second major pathway for the disposition of paraldehyde was its excretion in expired air, which, at infinite time, would account for 9.6% of the dose. No acetaldehyde (AcH) could be detected in either the breath or the blood of mice after paraldehyde administration. Pretreatment with the aldehyde dehydrogenase inhibitors, pargyline or cyanamide, did not result in the accumulation or excretion of detectable amounts of AcH. Pretreatment of mice administered [14C]paraldehyde with both cyanamide and D penicillamine,, an AcH sequestering agent, resulted, however, in urinary excretion of the 14C-labeled condensate of D-penicillamine and AcH showing AcH to be formed from paraldehyde. The above results indicate that paraldehyde is rapidly metabolized in vivo to carbon dioxide and that AcH is an intermediary product in this process. PMID- 6298406 TI - Functional evidence for two stereochemically different alpha-2 adrenoceptors regulating central norepinephrine and serotonin release. AB - The effects of norepinephrine and of various alpha adrenoceptor antagonists on the depolarization-evoked release of norepinephrine and 5-hydroxytryptamine were studied in nerve terminals isolated from rat cerebral cortex, preincubated with the radioactive amines and exposed during superfusion to 15 mM KCI. Exogenous norepinephrine inhibited in a concentration-dependent way both the release of norepinephrine and that of serotonin. The inhibitory effect of norepinephrine was antagonized by yohimbine and mianserin, but not by prazosin, indicating the involvement of alpha-2 adrenoceptors. The two enantiomers of mianserin were examined as alpha-2 adrenoceptor antagonists in the two release systems. Only (+) mianserin was an effective antagonist at the alpha-2 autoreceptors mediating regulation of norepinephrine release; (-)-mianserin was inactive. In contrast, both enantiomers antagonized exogenous norepinephrine at the alpha-2 adrenoceptors regulating 5-hydroxytryptamine release. It can be concluded that the alpha-2 adrenoceptors which regulate, respectively, norepinephrine and serotonin release in the cerebral cortex are located on presynaptic nerve terminals and represent two stereochemically different subtypes of alpha-2 adrenoceptors. PMID- 6298407 TI - Some characters of vacuum-uv effects on Sendai virus. PMID- 6298408 TI - Presence of an endonuclease specific for ultraviolet light-irradiated deoxyribonucleic acid in a ultraviolet light-sensitive mouse cell mutant Q31. PMID- 6298409 TI - Functional and biochemical heterogeneity among subpopulations of rat and mouse peritoneal macrophages. PMID- 6298410 TI - Characterization of the mannose/fucose receptor on human mononuclear phagocytes. AB - Cells of the mononuclear phagocyte system contain a cell surface receptor which mediates the uptake of mannose- and fucose-terminated glycoproteins. We have extended the initial studies to human alveolar and monocyte-derived macrophages in culture using two radiolabelled ligands, the synthetic glycoconjugate mannose bovine serum albumin and the lysosomal glycosidase, beta-glucuronidase. Uptake (37 degrees C) of 125I-mannose-BSA by freshly isolated alveolar macrophages is saturable with increasing concentrations of ligand. Kuptake values in macrophages of smokers and nonsmokers are similar, and resemble earlier reported values using rabbit alveolar macrophages (Kuptake = 40 nM). Uptake of 125I-mannose-BSA in cultured smoker macrophages is identical to that found in fresh cells, and uptake is stable for 5-10 days in culture. Fucose- and mannose-BSA are the most effective inhibitors of uptake, while N-acetylglucosamine-BSA is inhibitory at slightly higher concentrations. Binding (4 degrees C) of 125I-mannose-BSA is likewise ligand concentration dependent (KD = 30 nM). Freshly isolated human monocytes from healthy subjects and patients with cystic fibrosis do not have mannose-specific uptake. However, after monocytes are in culture for 3 days, mannose-specific uptake appears and Kuptake values and specificity of uptake are identical with the results from the alveolar macrophages. No uptake of mannose BSA could be found in the human monocyte-like cell line, U937. PMID- 6298411 TI - The effects of silica and fly ash dust inhalation on alveolar macrophage effector cell function. AB - Inhalation of silica was found to cause significant enhancement of alveolar macrophage antibody-dependent cell-mediated cytotoxicity (ADCC) after 14, 42, and 70 days of exposure, while similar treatment using fly ash resulted in significant suppression of ADCC after 42 days of exposure. Both silica and fly ash inhalation depressed the ability of alveolar macrophages from BCG-primed and BCG-rechallenged animals to mediate tumor cell lysis. Fly ash exposure also significantly suppressed the ability of BCG-activated macrophages to lyse target cells by the ADCC mechanism. PMID- 6298413 TI - Analytical subcellular fractionation of guinea pig peritoneal macrophages: preparation of purified plasma membranes. AB - A method using sucrose gradient centrifugation is described for the purification of plasma membranes of guinea pig peritoneal macrophages. The subcellular fractions obtained have been submitted to a biochemical and ultrastructural analysis. Two plasma membrane markers, 5'-nucleotidase and alkaline phosphodiesterase I, have been assayed at the same time as markers for other subcellular organelles, DNA (nuclei), succinic dehydrogenase (mitochondria), inosine diphosphatase (endoplasmic reticulum), and acid phosphatase (lysosomes). The exposure of the plasma membranes to a low concentration of digitonin allowed us to obtain their high purification. They are only contaminated by 2-3% of other cell components present in the macrophages homogenate. The representative ultrastructural technique used has confirmed the purity of the plasma membranes isolated. PMID- 6298412 TI - Separation of bronchoalveolar cells from the guinea pig on continuous density gradients of Percoll: morphology and cytochemical properties of fractionated lung macrophages. AB - Cells recovered by lavage from lungs of normal guinea pigs were centrifuged on continuous density gradients of colloidal silica (Percoll). The gradient was divided into six fractions based on the banding pattern of cells. This pattern was highly reproducible from animal to animal. Cell types in the fractions were identified by morphological and cytochemical criteria and the volume of the cells was determined by measuring their diameter and tritiated water space. More than 70% of the cells put on the gradient were recovered in the six fractions and there was no selective loss of cell types. Macrophages comprised more than 95% of the cells in fractions 3, 4, and 5. These fractions were of intermediate density (1.037-1.078 gm/ml) and together contained more than 85% of the recovered macrophages. Fraction 6 (density 1.078-1.130 gm/ml) was enriched for lymphocytes and granulocytes. Macrophages in fraction 5 were smaller, had more densely staining cytoplasm, and exhibited more nonspecific cytoplasmic esterase activity than macrophages in other fractions (5 greater than 4 greater than 3 greater than 2). These results indicate that density-gradient centrifugation on Percoll is an efficient method for purifying guinea pig alveolar macrophages and demonstrate that macrophages that differ in bouyant density also differ in morphologic and cytochemical properties. In a companion paper we report that macrophages in fractions 3, 4, and 5 differ functionally as well [9]. PMID- 6298415 TI - A study of the combined effect of ACTH(gel) and D-penicillamine on the functional disability of patients with rheumatoid disease. AB - Over a 6-year period, in 48 patients given concomitant D-penicillamine and ACTH(gel), the rapid induction of remission of the rheumatoid disease as judged by return to work in full and gainful employment was achieved in 77% within 1 month, in 71% by the end of 3 years, by which time 89% had achieved either Steinbrocker functional class I or II. Side effects were strikingly low in our patients with this regime, except for nephrotoxicity (10%); skin rash, dysgeusia and thrombocytopenia in 1 patient only and all patients were free of any gastrointestinal upset. Subsequent treatment of the failures with gold produced an overall remission rate of 90% in 3 years. PMID- 6298414 TI - Separation of bronchoalveolar cells from the guinea pig on continuous gradients of Percoll: functional properties of fractionated lung macrophages. AB - Lung macrophages from normal guinea pig lungs were separated from bronchoalveolar cells into three fractions according to buoyant density by centrifugation on continuous iso-osmotic gradients of Percoll [3]. A reproducible pattern of functional activity distinguished these three macrophage fractions. With increasing density and decreasing cell size, the respective fractions exhibited increased stimulated migration, superoxide anion release and pinocytosis, and increased protein concentration of the cells. These differences, coupled with previous observations that these fractions also exhibited morphological and cytochemical differences [3], support the notion that these fractions of macrophages may represent different stages of maturation (or differentiation) of alveolar macrophages in the lungs of normal guinea pigs. PMID- 6298416 TI - Progressive systemic sclerosis and pulmonary malignancy. PMID- 6298417 TI - Microelectrode exploration of human nerves: physiological and clinical implications. PMID- 6298419 TI - Antiviral compounds. 1. Structure-activity relationship of some antiviral enediones derived from aldehydo sugars. AB - A series of aldehydo sugars was subjected to condensation reactions with active methylene compounds. Acetylacetone was condensed with 2,4-O-benzylidene-3,5-O dibenzoyl-D-ribose (1), 2,4:3,5-O-dibenzylidene-D-ribose (6), 2,3,4,5-tetraacetyl D-ribose (7), and 2,3,4,5,6-pentaacetyl-D-glucose (9) to yield 3-ylidene-2,4 pentanedione derivatives 2, 11, 12, and 13, respectively. Sugar derivatives 1 and 6 were also condensed with benzoylacetone to give 14 and 18, with acetoacetanilide to give 16 and 19, with malononitrile to give 17 and 20, and with alpha-(gamma-butyrolactonylidene)triphenylphosphorane to give 21 and 22, respectively. Condensation of 1 with dibenzoylmethane gave 15. The double bond in compounds 2 and 11 was saturated by hydrogenation to give 23 and 24. All alpha, beta-unsaturated carbonyl compounds obtained exhibited antiviral activity and cytotoxicity. Compound 11 was found to have the most significant and selective antiviral activity against herpes simplex virus. PMID- 6298418 TI - Iatrogenic neuromuscular disorders: a review. PMID- 6298420 TI - New carboxyalkyl inhibitors of brain enkephalinase: synthesis, biological activity, and analgesic properties. AB - New carboxyalkyl compounds derived from Phe-Leu and Phe-Ala were synthesized and checked as inhibitors of "enkephalinase", a metalloendopeptidase cleaving the Gly3-Phe4 bond of enkephalins from mouse striatal membranes. Differential recognition of both brain enkephalinase and angiotensin-converting enzyme (ACE) catalytic sites by these carboxylalkyl compounds lead to potent (KI approximately 0.5 microM), competitive and selective inhibitors of the enkephalin-degrading enzyme. The most interesting compound, N-[(RS)-2-carboxy-3-phenylpropanoyl]-L leucine (3, KI = 0.34 microM), is 10000 times more potent on enkephalinase than on ACE activities. Intracerebroventricular (icv) injection of 3 in mice leads to a high potentiation of the analgesic effect of the exogenously administered D Ala2-Met-enkephalin, evidencing the in vivo inhibition of enkephalinase. Moreover, icv administration of 3 alone induces a dose-dependent analgesia in mice measured on both hot-plate and writhing tests. In the former assay, the ED50 was approximately 10 micrograms per animal, slightly higher than that of thiorphan. All the antinociceptive effects were antagonized by naloxone, demonstrating the involvement of enkephalins in analgesia and their in vivo protection from enkephalinase by 3. The described compounds can be considered as first examples of a new series of analgesics and potentially psychoactive agents. PMID- 6298421 TI - Synthesis and 5-lipoxygenase inhibitory activities of eicosanoid compounds. AB - Ten eicosanoid compounds (3, 6, 9, 11, 12, 15, 18, 21, 23, and 25), methyl (6E,8Z,11Z,14Z)-5-hydroxy-6,8,11,14-eicosatetraenoate (5-HETE, 10), leukotriene A4 (26), and (5S,6E,8E,10E,12RS,14E)-5,12-dihydroxy-6,8,10,14-eicosatetraenoic acid (5,12-diHETE, 27) were prepared and their inhibitory activities against the 5-lipoxygenase from guinea pig polymorphonuclear leukocytes (PMNL) were tested. 5,6-Methanoleukotriene A4 (18) was especially a potent and specific inhibitor of the 5-lipoxygenase without inhibiting the cyclooxygenase and the 12-lipoxygenase. Leukotriene A4, 5-HETE, and 5,12-diHETE also have inhibitory activities against the 5-lipoxygenase at micromolar concentrations, which can regulate the formation of slow-reacting substance of anaphylaxis intracellulary. PMID- 6298422 TI - Nucleosides. 123. Synthesis of antiviral nucleosides: 5-substituted 1-(2-deoxy-2 halogeno-beta-D-arabinofuranosyl)cytosines and -uracils. Some structure-activity relationships. AB - The syntheses of several 2'-halogeno-5-substituted-arabinofuranosylcytosines and uracils are described, and relationships of structure to anti herpes virus activity in vitro were examined. Those arabinonucleosides containing the 2' fluoro function exhibit, generally, more potent anti herpes virus (HSV) activity than do their 2'-chloro of 2'-bromo analogues. The importance of the fluorine in the 2'-"up" (arabino) configuration for enhancement of antiviral effectiveness is demonstrated by the superior activity of 2'-fluoro-5-iodo-ara-C [3a, FIAC] to that of 2'-fluoro-5-iodo-ribo-C. Of all the nucleosides tested herein, FIAC exhibited the most potent in vitro activity against HSV. 2'-Chloro-5-iodo- and -5 methyl-ara-C (3b and 4b) were 37 to greater than 500 times more effective in vitro against HSV type 2 than against type 1, suggesting that these latter derivatives might serve clinically as useful probes to distinguish between HSV types 1 and 2 in the diagnosis of HSV infections in man. PMID- 6298423 TI - Carbocyclic analogues of 5-substituted uracil nucleosides: synthesis and antiviral activity. AB - Carbocyclic analogues of 3'-deoxyuridines, 3'-deoxyuridines, and uridines with substituents at position 5 of the uracil moiety were prepared by direct halogenation (5-bromo and 5-iodo groups) and by displacement of the 5-bromo group by amino and substituted-amino groups. The analogue of 5-(hydroxymethyl)uridine was prepared via reaction of the isopropylidene derivative of the uridine analogue with paraformaldehyde. The carbocyclic analogues of thymidine and of 5 bromo-, 5-iodo-, and 5-(methylamino)-2'-deoxyuridine were highly active in vitro against herpes simplex virus, types 1 and 2. The corresponding analogues of 5 substituted 3'-deoxyuridines and of 5-substituted uridines were not active in this assay. PMID- 6298424 TI - 1-(4-Aminobenzyl)-and 1-(4-aminophenyl)isoquinoline derivatives: synthesis and evaluation as potential irreversible cyclic nucleotide phosphodiesterase inhibitors. AB - In an effort to increase the specificity of the potent phosphodiesterase inhibitor papaverine, we synthesized two series of novel 1-(4-aminobenzyl)- and 1 (4-aminophenyl)isoquinoline derivatives, incorporating alkylating moieties on the amine substituents. These compounds were evaluated for their inhibitory action on phosphodiesterase preparations from bovine heart and rat cerebral cortex. Studies were also conducted to determine whether these compounds were reacting with the enzymes in an irreversible manner. The compounds were potent inhibitors of the phosphodiesterases; however, no evidence was found for an irreversible inhibition. PMID- 6298425 TI - Synthesis of esters of phosphonoformic acid and their antiherpes activity. AB - Aliphatic and aromatic mono-, di-, and triesters of phosphonoformic acid (foscarnet) were synthesized. The triesters were prepared by the Michaelis Arbuzov reaction and were hydrolyzed to di- and monoesters. The compounds were tested for antiviral activity on isolated herpes simplex virus type 1 (HSV-1) DNA polymerase, in a HSV-1 plaque reduction assay, and on a cutaneous HSV-1 infection in guinea pigs. None of the esters inhibited the activity of isolated HSV-1 polymerases. Monoesters with a free carboxylic group and diesters with an aromatic carboxylic ester function were active against the cutaneous herpes infection. Mono- and diesters with an aromatic phosphonic ester group also showed activity in the plaque-reduction assay. However, mono- and diesters with an aromatic phosphonic ester group also showed activity in the plaque-reduction assay. However, mono- and diesters with aliphatic carboxylic ester groups were inactive in all test systems. The results show that all three acidic groups of phosphonoformic acid must be free in order to get antiviral activity at the enzyme level. However, certain esters of this acid may be biotransformed to the acid itself to give antiherpes activity. PMID- 6298426 TI - Piperazinylimidazo[1,2-a]pyrazines with selective affinity for in vitro alpha adrenergic receptor subtypes. AB - Regioselective syntheses of alkyl- and halogen-substituted piperazinylimidazo[1,2 a]pyrazines by novel oxidation-dehydration of [(beta-hydroxyalkyl)amino]pyrazines are described. Lanthanide shift reagent studies allowed correction of literature assignments of NMR chemical shifts and coupling constants for the imidazo[1,2 a]pyrazine ring system (e.g., J5,8 greater than J6,8). Equilibrium constants for displacement of specifically bound [3H]clonidine and [3H]prazosin from calf cerebral cortex homogenates in vitro are tabulated for reference and title compounds, and structure-affinity relationships for alpha 2- vs. alpha 1 adrenergic receptors are considered. Compound 2a, 8-(1-piperazinyl)imidazo[1,2 a]pyrazine, is equipotent with mianserin on the clonidine receptor (alpha 2) but ca. 70 times as selective as mianserin for this alpha 2-adrenergic receptor. Reduction of the imidazo ring (2,3-dihydro) lowers affinity for the alpha 2 receptor without affecting alpha 1-receptor affinity. Computer-assisted molecular modeling techniques are applied to the estimation of conformational energies of 2a and its 5-position isomer in relation to the semirigid molecule mianserin. PMID- 6298427 TI - Synthesis of (7R)-7H-indolo[3,4-gh][1,4]benzoxazines, a new class of D heteroergolines with dopamine agonist activity. AB - Synthesis of several members of the 9-oxaergoline ring system is presented. Both the C/D cis and the C/D trans isomers of 4,6,6a,8,9,10a-hexahydro-7-ethyl-7H indolo[3,4-gh] [1,4]benzoxazine were prepared, and the C/D trans isomer was resolved into its optical isomers. The enantiomer having the highest affinity for the [3H]apomorphine binding site, (-)-trans-6-ethyl-9-oxaergoline [(-)-6b], was shown to have the same absolute configuration as the natural ergolines, namely, 6aR, 10aR. In vivo and in vitro pharmacological evaluation shows these 9 oxaergolines to possess potent dopamine agonist properties. PMID- 6298428 TI - Antihypertensive agents: angiotensin converting enzyme inhibitors. 1-[3 (Acylthio)-3-aroylpropionyl]-L-prolines. AB - A series of 1-[3-(acylthio)-3-aroylpropionyl]-L-proline derivatives was synthesized. A number of these compounds are potent angiotensin converting enzyme (ACE) inhibitors that lowered blood pressure in aorta-coarcted renal hypertensive rats. The most active derivatives are 1-[3(R)-(acetylthio) -3-substituted benzoyl)-2(S)-methyl-propionyl]-L-prolines with an in vivo activity equivalent to SQ 14,225 (captopril). Structure-activity relationships are discussed. Changes in the configuration of the alpha-methyl group and the S-acetyl group affect the ACE activity. Coupling of 3-(substituted-benzoyl)-2-methylpropionic acids to L proline via enol lactones is described. PMID- 6298429 TI - (Mercaptopropanoyl)indoline-2-carboxylic acids and related compounds as potent angiotensin converting enzyme inhibitors and antihypertensive agents. AB - 1-(3-Mercapto-2-methyl-1-oxopropyl)indoline-2-carboxylic acids (7b) and related compounds were synthesized in order to examine their ability to inhibit angiotensin converting enzyme (ACE) and to reduce the systolic blood pressure of spontaneously hypertensive rats (SHR). All four possible stereoisomers of the precursor 1-[3-(benzoylthio)-2-methyl-1-oxopropyl]indoline-2-carboxylic acid (6b) were characterized with absolute stereochemical assignment. The removal of the benzoyl group of the precursor to give 7b was conveniently carried out by treatment with 2-methoxyethylamine. Three of the four stereoisomers of the benzoyl derivative 6 showed in vitro ACE inhibitory activity in the following order: 6b(S,S) greater than 6b(S,R) greater than 6b(R,S). The stereoisomer having the R,R configuration was essentially inactive. The substitution at the C5 of the indoline nucleus with the Et or OMe group caused only marginal changes in the inhibitory activity. The mercaptan 7b(S,S) was the most active ACE inhibitor synthesized in this study, showing in vitro potency 3 times that of captopril. The augmentation of the potency may be due to the increased hydrophobicity of 7b(S,S) compared with captopril and suggests the presence of a hydrophobic pocket at the active site of ACE. When tested in spontaneously hypertensive rats, 7b(S,S) exhibited oral antihypertensive activity 27 times that of captopril. The corresponding benzoyl derivative 6b(S,S) was 24 times as potent as captopril. The thio lactone 10 obtained by cyclization of 7b(S,S) as a potential prodrug was less potent than the parent compound, 7b(S,S), in the ACE inhibitory and antihypertensive tests. PMID- 6298430 TI - Synthesis of (E)-1-(5-chlorothien-2-yl)-2-(1H-imidazol-1-yl)ethanone 2,6 dichlorophenylhydrazone hydrochloride, a novel, orally active antifungal agent. AB - The preparation, determination of isomeric configuration, and antifungal properties of (E)-1-(5-chlorothien-2-yl)-2-(1H-imidazol-1-yl)ethanone 2,6 dichlorophenylhydrazone hydrochloride (1) are described. In vitro, compound 1 has been shown to have activity against Candida albicans comparable with miconazole. When administered orally to animals with experimentally induced vaginal candidiasis or systemic candidiasis, compound 1 produced results approaching those produced by ketoconazole. In addition, topical administration of compound 1 to rats with vaginal candidiasis produced results comparable with those produced by similar administration of clotrimazole. Unlike ketoconazole, which is active by a mechanism that is essentially fungistatic, compound 1 shares with miconazole a mode of action that is fungicidal. However, unlike miconazole, compound 1 exhibits activity following oral administration. Compound 1 has been found to be negative in the Ames test. PMID- 6298431 TI - Comparative arboviral susceptibility of female Culex tarsalis (Diptera: Culicidae) collected in CO2-baited traps and reared from field-collected pupae. PMID- 6298432 TI - Evolutionary changes of nucleotide sequences of papova viruses BKV and SV40: they are possibly hybrids. AB - Complete nucleotide sequences were compared between papova viruses BKV and SV40 and the degrees of sequence divergences were compared between structurally and/or functionally different segments or genes in details. It was shown that the rate of synonymous substitution is not only very high but also approximately uniform among different genes in these viruses as in eukaryotic genes examined to date. While all the non-coding regions including the intron showed marked sequence preservation which is in sharp contrasted with the case of eukaryotic genes where the large bulk of non-coding regions evolve at a rate as rapidly as that of synonymous substitution. It is remarkable that a long continuous stretch of sequence including the putative VPX gene and a 5' half of VP2 gene showed strong homology between BKV and SV40. A close examination of the pattern of base substitutions revealed that this unusual homology was derived by recombination between the two viruses during their evolution. On the basis of the pattern of base substitutions and the bias in code word utilization, we also showed that the putative VPX gene actually could code for a functional polypeptide. In papova viruses, the 3' terminal sequence of VP2/3 gene overlaps with the 5' terminal sequence of VPI gene. The pattern of base substitutions in the overlapping segment was examined in detail in comparison with those in the non-overlapping portions of VP2/3 and VP1 genes. It was shown that the evolutionary mode of the overlapping genes is in good agreement with our previous prediction. PMID- 6298433 TI - Lead administration during pregnancy and lactation affects steroidogenesis and hormone receptors in testes of offspring. AB - Sprague-Dawley rats were injected sc with lead acetate in suspension, or with sodium acetate (controls) on the d 9 of pregnancy and every 3-4 d thereafter until the pups were 13 or 21 d of age. At termination, testicular homogenates or isolated Sertoli cells were used to study steroidogenesis and gonadotropin binding. Lead had no significant effect on the mother's water and food consumption, on the pup's body or testis weights, on the number of pups and the time of birth, and on the seminiferous tubule diameter. Homogenates of testes of the lead-treated group converted significantly less (p less than 0.01) labeled progesterone (14C or 3H) to 5 alpha-pregnane-3,20-dione, 3 alpha-hydroxy-5 alpha pregnan-20-one, 17 beta-hydroxy-5 alpha-androstan-3-one (DHT), 3(alpha, beta) hydroxy-5 alpha-androstan-17-one, testosterone/17 alpha-hydroxyprogesterone, and androstenedione. Sertoli cells from lead-treated animals converted significantly less (p less than 0.01) progesterone to 5 alpha-pregnane-3 alpha, 20 alpha-diol, 3 alpha-hydroxy-5 alpha-pregnan-20-one, DHT, and 20 alpha-hydroxy-4-pregnen-3 one. These data and direct spectrophotometric assays indicated that 3 alpha hydroxysteroid oxidoreductase (3 alpha-HSO), 3 beta-HSO, 20 alpha-HSO, 5 alpha reductase, and C17-20-lyase had been affected. The receptor studies showed that the binding of [125I]rFSH to testicular receptors was significantly reduced from 35,600 (control) to 25,980 cpm/mg protein (lead). This is the first evidence that lead exposure (in utero and via mother's milk) significantly reduces steroid production and hormone binding in the testis at the onset of puberty. PMID- 6298434 TI - Studies of organotin-Schiff base complexes as new potential amebicidal agents. PMID- 6298435 TI - Alterations in the humoral immune response and tumor frequencies in mice exposed to benzo[a]pyrene and X-rays before or after birth. PMID- 6298437 TI - Effects of thyroxine on transmembrane resting potentials of skeletal muscle cells in culture. AB - The effect of thyroxine (T4) was studied on the transmembrane resting potential (Em) of rat skeletal muscle cells in culture. In agreement with previous studies, we found the Em of developing muscle cells to increase with age from Day 3 to Day 7 or 8, at which time a plateau is reached. The values of Em we obtained (-65 to 75 mV) were 15-20 mV higher than those (-50 to -55 mV) reported by others. Treatment of mature myotubes with T4 resulted in a significant increase in resting Em. There was only a slight difference between T4-treated and control cells with regard to the relation between extracellular K+ concentration and resting Em. Within 20-30 min of exposure to ouabain (10(-4)--10(-3)M) resting Em of both T4-treated and control cells had fallen to about the same value. This effect was reversible. Thus, T4 stimulates the activity of an electrogenic Na/K ATPase in the membrane of cultured skeletal muscle cells. PMID- 6298436 TI - Toxicity of 3,3',4,4'- and 2,2',5,5'-tetrabromobiphenyl: correlation of activity with aryl hydrocarbon hydroxylase induction and lack of protection by antioxidants. AB - 3,3',4,4'-Tetrabromobiphenyl is a minor component of commercial polybrominated biphenyl (PBB) mixture fireMaster BP-6 and is a potent inducer of aryl hydrocarbon hydroxylase (AHH). A single ip dose of 3,3',4,4'-tetrabromobiphenyl (150 mumol/kg) caused significant reduction in the growth rate in the immature male Wistar rat, as well as pale enlarged livers and marked reduction in thymus size. Under light microscopy, hepatocytes were enlarged and vacuolated. The vacuoles, which were most prominent in the midzonal region of the lobule, corresponded to fat droplets in oil-red-O-stained sections. The thymus, especially the cortex, was markedly depleted of lymphocytes. Neither the reduced growth, altered organ weights nor the histopathology was reversed for the duration of the study by the coadministration of the antioxidants butylated hydroxy anisole (BHA), butylated hydroxy toluene (BHT), or vitamin E. Vitamin E did, however counter the negative effect of 3,3',4,4'-tetrabromobiphenyl on growth during the first 5 d of the study. 2,2',5,5'-Tetrabromobiphenyl, also a minor component of fireMaster BP-6, is a weak phenobarbital-type inducer of cytochrome P-450. When administered at the same dose, 2,2',5,5' tetrabromobiphenyl did not elicit any observed toxic effects. These data confirm the correlation between AHH induction and toxicity for these PBBs and suggest that 3,3',4,4'-tetrabromobiphenyl may significantly contribute to the toxicity of fireMaster BP-6. Although there is evidence that polychlorinated biphenyls, and perhaps 2,3,7,8-tetrachlorodibenzo-p-dioxin, exert certain toxic effects via a lipid peroxidation mechanism, the toxic changes measured during this study were not reversed by the administration of the antioxidants. PMID- 6298438 TI - Comparison of centrifugation and filtration assays of ligand binding: do multiple GABA receptive sites exist? AB - Filtration and microcentrifugation procedures for the separation of bound and free ligand are compared for the assay of GABA receptors. The 3H-GABA and 3H muscimol are employed as ligands in saturation analyses of SPM and JC preparations. A direct comparison of the two methods, applied to two membrane preparations and two ligands, reveals that nonspecific binding is consistently higher with the microcentrifugation procedure. Analysis of the binding data yields essentially the same constants in either case; however, the filtration assay provides a better estimate in all cases. PMID- 6298439 TI - Ventromedial nucleus of the hypothalamus: convergent excitatory and inhibitory responses to fimbria and stria terminalis stimulation. AB - Field and extracellular unitary potentials were recorded in the ventromedial nucleus of the hypothalamus (VMH) of urethane-anesthetized rabbits after stimulation of the fimbria and stria terminalis. Stimulation of the lateral portion of the fimbria, which carries fibers from the ventral subiculum of the hippocampal formation, evoked a two-component response. An early excitatory response, with an average latency of 10 msec, predominated along the lateral margins of the VMH. A later inhibitory potential with an average latency of 15 msec was seen predominantly within the central portions of the VMH. Stimulation of the dorsal component of the stria terminalis produced two similar response patterns: an early excitatory response with an average latency of 16 msec, followed by an inhibitory potential with an average latency of 25 msec. The topographical distribution of these two components of the response was nearly identical to that produced by lateral fimbria stimulation. In contrast, stimulation of the ventral component of the stria terminalis evoked a simple excitatory response with an average latency of 10 msec which was maximal within the core of the VMH. Extracellular unitary recordings showed that the early negativity associated with stimulation of each of these three pathways reflects a monosynaptic excitation of VMH cells and that there was convergence of the three excitatory inputs at the single cell level. PMID- 6298441 TI - Effect of transganglionic degenerative atrophy on opiate receptors in the dorsal horn of the spinal cord. AB - We investigated alterations in opiate-binding sites in the upper dorsal horn after transection of the related peripheral sensory nerve in rats. The binding of (3H)diprenorphine was measured autoradiographically. The findings indicated a shift of the binding sites, rather than a degenerating disappearance, with a decrease in nerve fibers but an increase in nerve cells. This may be due to latent opiate-binding sites becoming manifest. PMID- 6298440 TI - Uptake, release, and binding of taurine in degenerated rat retinas. AB - High-affinity uptake, potassium-stimulated release and receptor binding of gamma aminobutyric acid (GABA) and taurine were studied in retinas of rats treated with monosodium glutamate (MSG) or iodoacetate-malate (IAM). Such treatments produce a selective damage of the inner retinal layers (MSG) or of the photoreceptor layer (IAM). MSG treatment decreased GABA uptake by more than 60%. Also, the potassium stimulated release of this amino acid was markedly diminished (90%) in retinas lesioned by MSG. MSG treatment decreased 3H-GABA binding by 45%. Uptake, release or binding of GABA were unaffected by IAM-treatment. Taurine uptake was reduced by a similar degree by both MSG and IAM treatment, 57% and 38% respectively. Potassium-stimulated release of taurine was unchanged by MSG and 50% reduced by IAM. Both treatments reduced the spontaneous release of endogenous taurine. Binding of taurine to receptors in retinal membranes was practically unaffected by any of the treatments utilized. These results are consistent with a neurotransmitter action of GABA at the inner retinal layers, while they do not support a major role for taurine as a synaptic transmitter in retina. PMID- 6298442 TI - Advances in methods and techniques for the identification of xenobiotic conjugates. AB - This review discusses recent advances in methods and instrumentation that are potentially useful for the characterization of polar xenobiotic conjugates. Topics that are discussed include: fast atom bombardment, secondary ion, 252Cf plasma desorption, field desorption and direct chemical ionization mass spectrometry and, proton and carbon-13 nuclear magnetic resonance spectrometry. PMID- 6298443 TI - The role of hydrolases in insecticide metabolism and the toxicological significance of the metabolites. AB - The hydrolytic enzymes involved in insecticide metabolism are the phosphorotriester hydrolases, carboxylesterases, carboxylamidases and epoxide hydrolases. The phosphorotriester hydrolases (arylesterases and DFP-ases) catalyze the P-anhydride bond cleavage of the "leaving group", a major route of detoxication of organophosphates. Carboxylesterases hydrolyze the carboethoxy group of malathion and also have hydrolytic activity toward certain synthetic pyrethroids. Carboxylamidases are involved in the hydrolysis of amide-containing phosphorothionates, N-formyl metabolites and substituted fluoroacetamides. Epoxide hydrolases hydrate certain cyclodiene insecticides and are probably involved in the metabolism of some other insecticides. Overall, the hydrolysis of insecticides increases the polarity of the metabolites and decreases their biological activity. PMID- 6298444 TI - Mechanisms of superoxide radical-mediated toxicity. AB - Free radicals generated from metabolism of foreign compounds can have extremely detrimental consequences on cell function and survival. Due to their high reactivity, free radicals may potentially perturb a wide spectrum of important cellular macromolecules such as nucleic acids, proteins, lipids and polysaccharides. Recently, the toxicity of several xenobiotics has been suggested to be mediated by formation of free radicals derived from reduction of molecular oxygen, forming superoxide anion (O(2)) and hydroxyl radical (OH .). For example, the pulmonary toxicity of the bipyridylium herbicide paraquat has been attributed to an enzymatically catalyzed one-electron redox cycling of the parent molecule, resulting in generation of O(2). Examples of other compounds that are subject to redox cycling with associated O(2) formation are those agents containing quinone or aromatic nitro structural elements. An important aspect of free-radical mediated toxicity is that it is moderated by several cellular defense mechanisms including superoxide dismutase, catalase, glutathione peroxidase, vitamin E and reduced glutathione. Thus, toxicity mediated by free radical generation may not occur unless defense mechanisms are overwhelmed by radical production. PMID- 6298445 TI - Toxicological significance of dihydrodiol metabolites. AB - Dihydrodiols are often found as the major organic-extractable metabolites of various olefinic or aromatic xenobiotics in many biological samples. Studies on the chemistry of dihydrodiol metabolites have provided insight into the pharmacokinetic behavior and the mode of action of the parent compound. The toxicology of dihydrodiol is more complex than what can be deduced solely on the basis of diminished bioavailability of the epoxide precursor, and the increased hydrophilicity associated with the dihydrodiol moiety. Dihydrodiols can be intrinsically toxic and may even represent metabolically activated species. Some of the dihydrodiol metabolites may still retain sufficient lipophilic character to serve again as substrates for microsomal oxygenases. Because of the tremendous chemical and biological diversity that existed among the various dihydrodiols, more mechanistic studies are needed to examine the toxicological properties of these compounds. It may be premature to conclude dihydrodiol formation as purely a detoxification route for xenobioties. PMID- 6298446 TI - Attachment of rous sarcoma virus to the fibronectin matrix of infected chick embryo fibroblasts. AB - During the early period of Rous Sarcoma Virus release from infected chick embryo fibroblasts, virions in close association with the extracellular fibrillar matrix were observed. This association was best seen in preparations fixed and embedded in situ and cut parallel to the basal surface of the cells. Goniometric and immunoenzymatic studies showed that virions were indeed attached to the fibrillar matrix and that these fibrils contained fibronectin. Fibronectin can be detected also on the virion surface. PMID- 6298447 TI - Organization and expression of the immediate early genes of human cytomegalovirus. AB - The immediate early genes of human cytomegalovirus were characterized according to map location, RNA transcripts, and translation products. Three regions in the large unique component (0.709 to 0.751 map units) were transcribed in the presence of an inhibitor of protein synthesis (anisomycin). A single size class of polyadenylated mRNA, 1.95 kilobases (kb), was transcribed abundantly relative to the other size classes. The predominant 1.95-kb viral RNA was transcribed from right to left on the prototype arrangement of the viral genome and spanned a region of approximately 2.8 kb (0.739 to 0.751 map units). This mRNA codes for a 75,000-dalton protein that represents the predominant immediate early protein detected in infected cells. Immunoprecipitation of viral proteins synthesized in vitro as well as in vivo demonstrated that the predominant immediate early protein is synthesized as a protein of 75,000 daltons, but is presumably modified in vivo, resulting in a broad banding pattern ranging from 75,000 to 68,000 daltons. A different immediate early viral gene (0.732 to 0.739 map units) is transcribed from left to right at relatively low levels. The 3' ends of the above viral RNAs terminate at approximately 230 base pairs apart in the region of approximately 0.739 map units. Five RNA size classes ranging from 2.25 to 1.10 kb were detected, but the 1.75-kb and 1.40-kb RNA size classes were more abundant from this region. At least four minor proteins are coded by these mRNAs, with apparent molecular weights ranging from 56,000 to 16,500. Last, a 1.95-kb mRNA was transcribed from a third region (0.709 to 0.728 map units). This viral mRNA was present at relatively low concentration and coded for a minor protein of 68,000 daltons. Since immediate early gene expression of human cytomegalovirus is dominated by the synthesis of an mRNA from the region of 0.739 to 0.751 map units that codes for the predominant immediate early protein found in the infected cell, we hypothesize that this protein is the major regulatory protein influencing the switch from restricted to extensive transcription. PMID- 6298448 TI - Application of denatured, electrophoretically separated, and immobilized lysates of herpes simplex virus-infected cells for detection of monoclonal antibodies and for studies of the properties of viral proteins. AB - We report the use of herpes simplex virus type 1 (HSV-1)- and HSV-2-infected cell polypeptides (ICPs) separated by electrophoresis in polyacrylamide gels and transferred to nitrocellulose to (i) detect monoclonal antibodies to viral polypeptides and to (ii) study the properties of the proteins with the monoclonal antibodies. Our results were as follows. (i) When the antigens were electrophoretically separated in denaturing gels and then immobilized on nitrocellulose strips, we detected a greater diversity of monoclonal antibodies to viral proteins than when we used the technique of immune precipitation of soluble, nondenatured viral antigens. The primary advantage of the technique is in the detection of nonprecipitating antibody and of antibody to poorly soluble antigens not available for reaction in preparations cleared by high-speed centrifugation before immune reaction. (ii) Studies of the viral polypeptides reactive with three monoclonal antibodies indicated that the technique can be used to investigate several properties of the antigens. Specifically, monoclonal antibody to ICP 4 confirmed the accumulation of viral protein in the nucleus and the mapping of the gene in the S component. The results showed, however, that HSV 1 and HSV-2 ICP 4 do have common antigenic determinants. The reaction of a nonprecipitating monoclonal antibody with electrophoretically separated, immobilized polypeptides contained in cytoplasmic and nuclear fractions, those chemically deglycosylated, or those specified by specific HSV-1 x HSV-2 intertypic recombinants identified the antigens reactive with the second monoclonal antibody as various forms of glycoprotein gC. Of particular interest was a set of four antigens, 39,000 to 46,500 in apparent molecular weight, reactive with each of several monoclonal antibodies. These studies showed that two polypeptides partition in the cytoplasm and two in the nucleus and that all comap with the previously mapped ICPs 35 and 37 in the region of the genome defined by the viral thymidine kinase gene on the left and the glycoprotein gA/B gene on the right. Unlike ICP 4 and gC, the four polypeptides are linked by intermolecular bisulfide bonds, inasmuch as the polypeptides were not at the expected locations upon denaturation and electrophoresis in the absence of reducing agents. PMID- 6298449 TI - Dimethyl sulfoxide affects the amount of extrachromosomal spleen focus-forming virus DNA in murine erythroleukemia cells. AB - We used Southern blot hybridization to titrate and map restriction enzyme cleavage sites of a 6.3-kilobase-pair species of extrachromosomal viral DNA found in derivatives of the 745A line of murine erythroleukemia cells, which vary in their ability to be induced to differentiate by dimethyl sulfoxide (DMSO). Greater than an eightfold variation was observed in the amount of this DNA, with the largest amounts being found in cells that were resistant to the induction of differentiation by DMSO. This increase in the level of extrachromosomal viral DNA was found to be dependent upon the continued presence of DMSO in the culture medium. The increase was shown not to be due to an immediate stimulatory effect of this agent on the synthesis or maintenance of this DNA, since cell lines sensitive to the differentiation-inducing effects of DMSO were shown to undergo a transient reduction in the amount of extrachromosomal viral DNA after the addition of DMSO to the culture medium. In addition to the 6.3-kilobase-pair linear form found in the cytoplasm, in some preparations two hybridizing bands were observed that migrated in agarose gels in the position expected of covalently closed circular species of viral DNA. Restriction enzyme mapping of the cytoplasmic linear form indicated a close relationship of this DNA to two polycythemic strains of spleen focus-forming virus that have been molecularly cloned by other workers. No obvious change in the number or arrangement of chromosomal viral sequences could be detected after treating cells with DMSO. Thus, the exposure of murine erythroleukemia cells to DMSO caused an obvious change in the amount of extrachromosomal spleen focus-forming virus DNA but no obvious change in the integration of the provirus. PMID- 6298450 TI - Sequence determination of the (+) leader RNA regions of the vesicular stomatitis virus Chandipura, Cocal, and Piry serotype genomes. AB - Using 3'-end-labeled genome probes, cells infected with vesicular stomatitis virus Chandipura, Cocal, and Piry serotypes were shown to contain (+) leader RNAs of approximately 50 nucleotides in length. The nucleotide sequence of the leader RNA regions of these genomes was determined and compared with the previously reported sequences of both the (+) and (-) leader RNA regions of other vesicular stomatitis virus serotypes. Regions of strong conservation of nucleotide sequence among the various vesicular stomatitis virus serotypes suggest those nucleotides thought to be involved in control functions during vesicular stomatitis virus replication. PMID- 6298451 TI - Topography of simian virus 40 A protein-DNA complexes: arrangement of pentanucleotide interaction sites at the origin of replication. AB - Investigation of the DNA binding properties of the simian virus 40 (SV40) A protein (large T antigen) and the hybrid adenovirus-SV40 D2 protein revealed that both viral proteins protect similar regions of SV40 DNA from digestion by DNase I or methylation by dimethyl sulfate. However, the interaction of D2 protein with DNA was more sensitive to increases of NaCl concentration than was the interaction of wild-type SV40 A protein. Dimethylsulfate footprinting identified 13 DNA pentanucleotide contact sites at the viral origin of replication. The sequences of these sites corresponded to the consensus family 5'-(G greater than T) (A greater than G)GGC-3'. The pentanucleotides were distributed in three regions of origin DNA. Region I contained three pentanucleotide contact sites arranged as direct repetitions encompassing a span of 23 base pairs. In region II, four pentanucleotides were oriented as inverted repetitions that also spanned a total of 23 base pairs. Region III had six recognition pentanucleotides arranged as direct repetitions in a space of 59 base pairs. These fundamental variations in DNA arrangement are likely to determine different patterns of protein binding in each region. PMID- 6298452 TI - Topography of simian virus 40 A protein-DNA complexes: arrangement of protein bound to the origin of replication. AB - DNA binding regions I, II, and III at the origin of replication have different arrangements of A protein (T antigen) recognition pentanucleotides. The A protein also protects each region from DNase in distinctly different patterns. Footprint and fragment assays led to the following conclusions: (i) in some cases a single recognition pentanucleotide is sufficient to direct the binding and accurate alignment of A protein on DNA; (ii) the A protein binds within isolated region I or II in a sequential process leading to multiple overlapping areas of DNase protection within each region; and (iii) the 23-base pair span of recognition sequences in region II allows binding and protection of a longer length of DNA than the 23-base pair span in region I. We propose a model of protein binding that addresses the problem of variations in the arrangement of pentanucleotides in regions I and II and explains the observed DNase protection patterns. The central feature of the model requires each protomer of A protein to bind to a pentanucleotide in a unique direction. The resulting orientation of protein would protect more DNA at the 5' end of the 5'-GAGGC-3' recognition sequence than at the 3' end. The arrangement of multiple protomers at the origin of simian virus 40 replication is discussed. PMID- 6298453 TI - Nucleotide sequence of a cDNA clone encoding the entire glycoprotein from the New Jersey serotype of vesicular stomatitis virus. AB - The nucleotide sequence of the mRNA encoding the glycoprotein from the New Jersey serotype of vesicular stomatitis virus (VSV) was determined from a cDNA clone containing the entire coding region. The sequence of 12 5'-terminal noncoding nucleotides present in the mRNA but not in the cDNA clone was determined from a primer extended to the 5' terminus of the mRNA. The mRNA is 1,573 nucleotides long (excluding polyadenylic acid) and encodes a protein of 517 amino acids. Only six nucleotides occur between the translation termination codon and the polyadenylic acid. Short homologies between the untranslated termini of this mRNA and the mRNAs of the Indiana serotype were found. The predicted protein sequence was compared with that of the glycoprotein of the Indiana serotype of VSV and with the glycoprotein of rabies virus, using a computer program which determines optimal alignment. An amino acid identity of 50.9% was found for the two VSV serotypes. Approximately 20% identity was found between the rabies virus and VSV New Jersey glycoproteins. The positions and sizes of the transmembrane domains, the signal sequences, and the glycosylation sites are identical in both VSV serotypes. Two of five serine residues which were possible esterification sites for palmitate in the glycoprotein from the Indiana serotype are changed to glycine residues in the glycoprotein from the New Jersey serotype. Because the glycoprotein of the New Jersey serotype does not contain esterified palmitate, we suggest that one or both of these residues are the probable esterification sites in the glycoprotein from the Indiana serotype. PMID- 6298455 TI - Structural and antigenic analysis of the nucleic acid-binding proteins of bovine and feline leukemia viruses. AB - The nucleic acid-binding proteins of bovine leukemia virus (BLV) and feline leukemia virus (FeLV) were isolated in a high state of purity with chloroform methanol extraction followed by reversed-phase liquid chromatography. Selective solubilization and purity of BLV p12 and FeLV p10 was confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The compositions and molecular weights were determined by amino acid analysis. An abundance of lysine and arginine residues along with their size identifies both BLV p12 and FeLV p10 as small basic proteins similar to well-defined type C viral nucleoproteins. NH2 terminal degradation by the semiautomated Edman method provided the sequence of the first 40 amino acids for both proteins. The putative nucleic acid binding site found in several type C viral nucleoproteins was contained within this sequence, with the most homology centered around an eight-amino acid region involving seven identical residues and one substitution. Antisera were developed in rabbits, and specificity and titers were determined by electroblotting and immunoautoradiography. By this technique, an immunological cross-reaction was found between BLV p12 and FeLV p10. The shared antigenic determinant most likely exists in the highly conserved eight-amino acid region. Although this sequence is also highly conserved in the nucleic acid-binding proteins of murine leukemia viruses, the shared antigenic determinant is not found in these or any other type C viruses tested. It is suggested that substitution of arginine (BLV p12/FeLV p10) to lysine (murine leukemia virus p10) is sufficient to elicit a change in antibody specificity. PMID- 6298454 TI - Nucleotide sequence of the region encompassing the JC virus origin of DNA replication. AB - The region of the JC virus (JCV) genome from 0.58 to 0.73 map units was sequenced by the Maxam-Gilbert technique. This segment of DNA specifies several regulatory elements and the amino-terminal portion of the early viral proteins. Comparisons with the analogous regions in the polyomaviruses simian virus 40 (SV40) and BK virus (BKV) confirm the close evolutionary relationship of these three viruses. Similarities include palindromic and symmetrical sequences near their origins of DNA replication, binding sites for their large T proteins, an AT-rich region (the Goldberg-Hogness, or TATA, box), and a large tandem duplication or triplication to the late side of their replication origins (however, these sequences differ). Homology between the sequences coding for the early proteins is also evident (79 and 93 of the first 110 amino acids are shared with SV40 and BKV, respectively). Of greater interest are features of the JCV genome which differ from those of other polyomaviruses. Absent in JCV and BKV are sequences which resemble the third T-antigen binding site of SV40. In addition, a set of sequences present in JCV and BKV DNAs (33 nucleotides in JCV and 22 nucleotides in BKV) and located near a 17-base-pair palindrome shared by all three viruses is missing in SV40 DNA. Another sequence, GGGXGGAG, which is repeated several times in many polyomaviruses and adenoviruses and which is thought to play a role in DNA replication or transcription or both, is not found near the JCV origin of replication. Finally, the tandem repeat of JCV, unlike those of BKV and SV40, includes the Goldberg-Hogness sequence. PMID- 6298456 TI - Effect of arabinofuranosylthymine on the replication of Epstein-Barr virus and relationship with a new induced thymidine kinase activity. AB - 1-beta-D-Arabinofuranosylthymine (araT) is a selective inhibitor of Epstein-Barr virus replication induced in both thymidine kinase (TK)-negative (TK-) and TK+ variants of the lymphoid cell line P3HR-I. This analog has no effect on the growth of noninduced cells (T. Ooka and A. Calender, Virology 104:219-223, 1980). The synthesis of early antigens is not affected by the analog, whereas that of late viral capsid antigens is completely inhibited, as demonstrated by the indirect immunofluorescence technique; kinetic reassociation experiments have also shown that araT strongly inhibits replication of viral DNA. Phosphorylation of the tritiated form of the analog ([3H]araT) was analyzed by thin-layer chromatography in cultures of control and induced cells, and the results demonstrated that only induced cells can convert the analog to the triphosphate form. These results indicate that the selective effect of araT in induced cells is probably related to a new virally induced TK activity. Preliminary characterization of this new activity has shown that it is able to phosphorylate the analog specifically, whereas cellular TKs cannot. araTTP, a final phosphorylation product of araT, is a potent inhibitor of Epstein-Barr virus specific DNA polymerase, suggesting a possible inhibitory action of this product on Epstein-Barr virus replication. PMID- 6298457 TI - Nucleotide sequence of the env-specific segment of NFS-Th-1 xenotropic murine leukemia virus. AB - The sequence of 863 contiguous nucleotides encompassing portions of the pol and env genes of NFS-Th-1 xenotropic proviral DNA was determined. This region of the xenotropic murine leukemia virus genome contains and env-specific segment that hybridizes exclusively to xenotropic and mink cell focus-forming but not to ecotropic proviral DNAs (C. E. Buckler et al., J. Virol. 41:228-236, 1982). The unique xenotropic env segment contained several characteristic deletions and insertions relative to the analogous region in AKR and Moloney ecotropic murine leukemia viruses. Portions of an endogenous env segment cloned from a BALB/c mouse embryo gene library that had a restriction map and hybridization properties typical of xenotropic viruses (A. S. Khan et al., J. Virol. 44:625-636, 1982) were also sequenced. The sequence of the endogenous env gene was very similar to the comparable region of the NFS-Th-1 xenotropic virus containing the characteristic deletions and insertions previously observed and could represent a segment of an endogenous xenotropic provirus. PMID- 6298458 TI - Structure and transcription of the cellular homolog (c-myb) of the avian myeloblastosis virus transforming gene (v-myb). AB - We isolated and characterized molecular clones containing the chicken cellular homolog (c-myb) of the avian myeloblastosis virus oncogene (v-myb). Mapping of the c-myb clones using restriction endonucleases and hybridization to radiolabeled v-myb probes revealed that the sequences homologous to v-myb are contained within four separate regions, which have since been shown by nucleotide sequencing (Klempnauer et al., Cell 31:453-463, 1982) to carry seven exons. Analysis of c-myb transcripts showed the presence of several large precursor RNAs in addition to the 4.0-kilobase cytoplasmic mRNA. We also determined the approximate positions of the c-myb sequences that are present in the 4.0-kilobase c-myb mRNA but not present in v-myb. Some of these sequences are found in a separate region 5' to the v-myb-related sequences, whereas the remainder appear to be located immediately 3' to the v-myb-related sequences. The data presented here, in conjunction with nucleotide sequence analysis (Klempnauer et al., Cell 31:453-463, 1982), indicate that the c-myb gene contains at least eight exons which span a total of about 16 kilobase pairs. The presence of exon sequences in c-myb outside the regions of homology with v-myb raises the possibility that the v-myb and c-myb gene products may differ significantly. PMID- 6298459 TI - Identification of a spleen focus-forming virus in erythroleukemic mice infected with a wild-mouse ecotropic murine leukemia virus. AB - An NFS/N mouse inoculated at birth with an ecotropic murine leukemia virus (MuLV) obtained from wild mice (Cas-Br-M MuLV) developed a lymphoma after 18 weeks. An extract prepared from the lymphomatous spleen was inoculated into newborn NFS/N mice, and these mice developed erythroleukemia within 9 weeks. Spleens from the erythroleukemic mice contained ecotropic and mink cell focus-inducing (MCF) MuLVs; however, when these viruses were biologically cloned and reinoculated into newborn NFS/N mice, no erythroleukemia was induced. In contrast, cell-free extracts prepared from the erythroleukemic spleens induced erythroleukemia within 5 weeks. Analysis of cell-free extracts prepared from the erythroleukemic spleens showed that they contained a viral species that induced splenomegaly and spleen focus formation in adult mice, with susceptibility controlled by alleles at the Fv-2 locus. The spleen focus-forming virus coded for a 50,000-dalton protein precipitated by antibodies specific to MCF virus gp70. RNA blot hybridization studies showed the genomic viral RNA to be 7.5 kilobases and to hybridize strongly to a xenotropic or MCF envelope-specific probe but not to hybridize with an ecotropic virus envelope-specific probe. The virus described here appears to be the fourth independent isolate of a MuLV with spleen focus-forming activity. PMID- 6298460 TI - Effects of tunicamycin on rotavirus morphogenesis and infectivity. AB - The functions of the two rotavirus glycoproteins were investigated by using tunicamycin and a variant of SA11 rotavirus having nonglycosylated VP7. Results showed that glycosylation of VP7 is not required for normal viral morphogenesis and infectivity and suggested that the nonstructural glycoprotein is involved in assembly of the outer capsid. PMID- 6298461 TI - Identification of a subset of herpesvirus saimiri polypeptides synthesized in the absence of virus DNA replication. AB - The effects of phosphonoacetic acid on the synthesis of herpesvirus saimiri specific polypeptides in productively infected cells were examined. At concentrations that inhibited virus DNA synthesis (greater than or equal to 150 micrograms/ml), phosphonoacetic acid prevented the synthesis of the majority of virus-specific polypeptides while allowing the synthesis of a subset of virus proteins (i.e., 110,000 [110K], 76K, 72K, 51K, 48K, 29K, 24K, and 20K or 21K) and the protracted synthesis of host-specified polypeptides. Other inhibitors of DNA synthesis (e.g., cytosine arabinoside) showed the same selective inhibition of late virus protein synthesis and identified the same resistant subset of early virus-specific polypeptides. This DNA synthesis-independent subset included the 51K phosphoprotein, which, together with the 110K, 48K, and 31K polypeptides, accumulated in the nuclear fraction of infected cells. PMID- 6298462 TI - Deletion mapping of a short polyoma virus middle T antigen segment important for transformation. AB - Viable polyoma virus mutants were constructed that had small deletions in the early region of the genome. The deletions together removed most of the segment missing from the genome of the nontransforming mutant dl23 (N. Smolar and B. E. Griffin, J. Virol. 38:958-967, 1981). The transformation properties, as measured by colony formation in soft agar, of mutants with overlapping or contiguous deletions showed that part or all of the middle T antigen segment, consisting of the short amino acid sequence Glu4-Tyr-Met-Pro-Met, was essential for the activity of the protein in transformation. However, the segment could be deleted without significant effect on the in vitro protein kinase activity associated with the middle T antigen. PMID- 6298464 TI - Glycosylation of herpes simplex virus type 1 gC in the presence of tunicamycin. AB - The presence of O-glycosidic linkages on herpes simplex virus type 1 (HSV-1) glycoproteins was indicated by the synthesis and glycosylation of HSV-1 glycoproteins in the presence of tunicamycin. Monospecific antiserum to HSV-1 gC immunoprecipitated a 92,000-molecular-weight protein synthesized in the presence of tunicamycin and isotopically labeled with glucosamine or galactose. Anti-gAB did not immunoprecipitate a carbohydrate-labeled HSV-1 protein synthesized in the presence of tunicamycin. The purified glucosamine-labeled 92,000-molecular-weight protein synthesized in the presence of tunicamycin and the fully glycosylated forms of gAB and gC were tested for their sensitivity to mild alkaline hydrolysis. Purified gAB was resistant to mild alkaline hydrolysis, whereas gC and the 92,000-molecular-weight protein were both sensitive to mild alkaline hydrolysis. These results suggest that O-glycosidic linkages are associated with the HSV-1 gC glycoprotein. PMID- 6298465 TI - Gene mapping of rotavirus double-stranded RNA segments by northern blot hybridization: application to segments 7, 8, and 9. AB - Cloned DNA copies of double-stranded RNA segments 7, 8, and 9 of UK bovine rotavirus were nick-translated with [alpha-(32)P]ATP and hybridized to double stranded RNA of various rotavirus strains which had been separated on long polyacrylamide gels and then transferred to o-aminophenylthioether paper. Specific hybridization of the UK calf clones to the separated RNA segments allowed the corresponding genes of four different rotaviruses to be rapidly determined. PMID- 6298463 TI - Mapping of multiple phosphorylation sites within the structural and catalytic domains of the Fujinami avian sarcoma virus transforming protein. AB - The phosphorylation sites of the P140gag-fps gene product of Fujinami avian sarcoma virus have been identified and localized to different regions of this transforming protein. FSV P140gag-fps isolated from transformed cells is phosphorylated on at least three distinct tyrosine residues and one serine residue, in addition to minor phosphorylation sites shared with Pr76gag. Partial proteolysis with virion protease p15 or with Staphylococcus aureus V8 protease has been used to generate defined peptide fragments of P140gag-fps and thus to map its phosphorylation sites. The amino-terminal gag-encoded region of P140gag fps contains a phosphotyrosine residue in addition to normal gag phosphorylation sites. The two major phosphotyrosine residues and the major phosphorserine residue are located in the carboxy-terminal portion of the fps-encoded region of P140gag-fps. P140gag-fps radiolabeled in vitro in an immune complex kinase reaction is phosphorylated at only one of the two C-terminal tyrosine residues phosphorylated in vivo and weakly phosphorylated at the gag-encoded tyrosine and at a tyrosine site not detectably phosphorylated in vivo. Thus, the in vitro tyrosine phosphorylation of P140gag-fps is distinct from that seen in the transformed cell. A comparative tryptic phosphopeptide analysis of the gag-fps proteins of three Fujinami avian sarcoma virus variants showed that the phosphotyrosine-containing peptides are invariant, and this high degree of sequence conservation suggests that these sites are functionally important or lie within important regions. The P105gag-fps transforming protein of PRCII avian sarcoma virus lacks one of the C-terminal phosphotyrosine sites found in Fujinami avian sarcoma virus P140gag-fps. Partial trypsin cleavage of FSV P140gag-fps immunoprecipitated with anti-gag serum releases C-terminal fragments of 45K and 29K from the immune complex that retain an associated tyrosine-specific protein kinase activity. This observation, and the localization of the major P140gag-fps phosphorylation sites to the C-terminal fps region, indicate that the kinase domain of P140gag-fps is located at its C terminus. The phosphorylation of P140gag-fps itself is complex, suggesting that it may itself interact with several protein kinases in the transformed cell. PMID- 6298466 TI - In vitro synthesis of the nonstructural C protein of Sendai virus. AB - In vitro translation was used to study the mRNA for the Sendai virus nonstructural C protein. The C protein mRNA was found to be coordinately expressed with the mRNAs for the structural proteins. However, the C protein mRNA appeared to be translated more efficiently in vitro than the other mRNAs. In addition, the 22,000-dalton C protein mRNA cosedimented on sucrose gradients with the 79,000-dalton P protein mRNA. The C protein mRNA thus appears to be much larger than expected. PMID- 6298467 TI - Sendai virus envelopes can mediate Epstein-Barr virus binding to and penetration into Epstein-Barr virus receptor-negative cells. AB - Epstein-Barr virus (EBV) receptor-negative cells were treated with UV-inactivated Sendai virus (SV) or with reconstituted SV envelopes having a low hemolytic activity and then assayed for EBV binding or for susceptibility to EBV infection. EBV binding was assessed by using both unlabeled and fluoresceinated EBV preparations. It was found that SV or SV envelope treatment renders these cells able to bind EBV. Various experiments were performed to clarify the mechanism of this SV-induced binding. The EBV receptor-negative 1301 cells were treated with SV either at 0 degrees C or at both 0 and 37 degrees C successively and then examined for EBV binding at 0 degrees C. It was thus found that when SV treatment was performed exclusively at 0 degrees C, the target cells showed higher fluorescence intensity after their incubation with fluoresceinated EBV. In addition, Clostridium perfringens neuraminidase treatment of 1301 cells did not induce any EBV binding to these cells. These data indicate that EBV binding is not due to the disturbance of the cell membrane by SV envelope fusion or to the uncovering of EBV binding sites on the cells after the enzymatic action of SV neuraminidase. Moreover, bound EBV was partly eluted from SV-treated 1301 cells at 37 degrees C, and the treatment of EBV with C. perfringens neuraminidase inhibited its SV-mediated binding. These data indicate that EBV binds to the hemagglutinin-neuraminidase of SV on the target cell surface and that a fraction of the bound EBV becomes irreversibly associated with the SV-treated cell membrane. Our data also show that EBV can penetrate into 1301 cells which have incorporated SV envelopes into their membrane, as demonstrated by the induction of the EBV-determined nuclear antigen by B95-8 EBV in SV envelope-treated 1301 cells. PMID- 6298468 TI - Alignment of the genome of monkey B-lymphotropic papovavirus to the genomes of simian virus 40 and BK virus. AB - We located the origin of DNA replication of African green monkey B-lymphotropic papovavirus DNA by analyzing pulse-labeled form I DNA. With the replication origin used as a reference point, the B-lymphotropic papovavirus genome was aligned with the genomes of simian virus 40 and BK virus from DNA homology between specific fragments hybridized under low-stringency conditions. From the results of these experiments, it was possible to deduce the correlation between the physical and functional maps of the B-lymphotropic papovavirus genome. PMID- 6298469 TI - Transcription of mouse mammary tumor virus: identification of a candidate mRNA for the long terminal repeat gene product. AB - We have examined an assortment of preneoplastic and neoplastic mouse mammary tissues for the presence of an mRNA which could encode the putative long terminal repeat gene product of mouse mammary tumor virus. We report here the detection of a novel mouse mammary tumor virus-specific, polyadenylic acid-containing transcript in certain preneoplastic and neoplastic mammary tissue of BALB/c mice. The molecule is 1.6 kilobases in length and contains sequences from the transcriptional leader and the U3 region of the proviral DNA. The upstream terminus of the 3' information lies 75 to 80 nucleotides from the beginning of the long terminal repeat open reading frame, in close proximity to a consensus splice acceptor in the DNA. The transcript was detected in hormonally or chemically induced neoplastic, preneoplastic, and lactating mammary tissue of BALB/c mice, but not in preneoplastic or tumor tissue induced by exogenous viruses in any strain of mice examined. This implies that the RNA we observed is transcribed from an endogenous provirus template. PMID- 6298470 TI - Alterations in the structure of the oligosaccharide of vesicular stomatitis virus G protein by swainsonine. AB - Swainsonine, an inhibitor of glycoprotein processing, inhibits the formation of the normal oligosaccharide chain of the G protein of vesicular stomatitis virus. Thus, when vesicular stomatitis virus was grown in baby hamster kidney cells in the presence of swainsonine (15 to 500 ng/ml) and labeled with [2-(3)H]mannose, the oligosaccharide portion of the G protein was completely susceptible to the action of endoglucosaminidase H. However, the normal viral glycoprotein is not susceptible to this enzyme. Various enzymatic treatments and methylation studies of the mannose-labeled oligosaccharides suggest that swainsonine causes the formation of a hybrid-type oligosaccharide having an oligomannosyl core (Man(5)GlcNAc(2)-Asn) characteristic of neutral oligosaccharides plus the branch structure (NeuNAc-Gal-GlcNAc) characteristic of the complex oligosaccharides. A structure for this hybrid oligosaccharide is proposed. Swainsonine had no effect on the incorporation of [(14)C]leucine into viral proteins, nor did it change the number of PFU produced in these cultures. It did, however, slightly decrease the incorporation of [(3)H]glucosamine and increase the incorporation of [(3)H]mannose. Vesicular stomatitis virus raised in the presence of swainsonine bound much more tightly to columns of concanavalin A-Sepharose than did control virus. Swainsonine had to be added within the first 4 or 5 h of virus infection to be effective. Thus, when 100 ng of the alkaloid per ml was added at any time within the first 3 h of infection, essentially all of the glycoprotein was susceptible to digestion by endoglucosaminidase H. However, when swainsonine was added 4 h after the start of infection, 30% of the glycopeptides became resistant to endoglucosaminidase H; at 5 h, 70% were resistant. The effect of swainsonine was reversible since removal of the alkaloid allowed the cells to form the normal complex glycoproteins. However, the time of removal was critical in terms of oligosaccharide structure. PMID- 6298471 TI - Somatically acquired recombinant murine leukemia proviruses in thymic leukemias of AKR/J mice. AB - We have probed the structure and arrangement of murine leukemia virus genomes in eight spontaneous AKR thymic leukemias by Southern hybridization with one ecotropic pol and four ecotropic env probes. These probes revealed many (in 2 cases over 15) somatically acquired proviruses that had undergone complex patterns of recombination. The large majority were not deleted and were structurally analogous to the oncogenic mink cell focus-inducing murine leukemia viruses isolated from AKR tumors in that the amino-terminal p15E-coding region derived from ecotropic AKR murine leukemia virus sequences, whereas certain gp70 coding sequences were nonecotropic. Nevertheless, we observed a few proviruses which did not appear to be gp70 recombinants; however, these proviruses were in general clearly recombinant within the p15E-coding sequences. Although the proviral recombination patterns were quite variable, in general the large majority of recombinant proviruses within each tumor appeared structurally identical, indicating that they originate from a common parent. Each tumor contained a unique pattern of provirus integrations; densitometer tracings of the Southern hybridizations indicated that many of the integrated proviruses were present at one copy per cell, suggesting that the tumors derive from a single cell which contained multiple integrated copies of a unique recombinant virus structurally similar to the mink cell focus-inducing viruses. PMID- 6298472 TI - Genetic variability of herpes simplex virus: development of a pathogenic variant during passaging of a nonpathogenic herpes simplex virus type 1 virus strain in mouse brain. AB - Herpes simplex virus type 1 ANG (HSV-1 ANG) is originally nonpathogenic for inbred mice upon intraperitoneal intravenous, or intravaginal inoculation. In contrast, mice died of encephalitis within 4 to 5 days after intracerebral inoculation with this strain. HSV-1 ANG was serially passaged in mouse brains. In two independent series, peripherally pathogenic virus variants had developed and accumulated in the virus progeny after 12 to 15 intracerebral passages. In mixed infections both nonpathogenic and pathogenic viruses replicated at the primary site of infection and spread to various organs. However, only the pathogenic phenotype could be recovered from the spinal cord and the brain. Comparison of the restriction enzyme cleavage patterns of pathogenic ANG and nonpathogenic ANG virus DNAs revealed distinct alterations in the S-segment (US) sequences bounded by coordinates 0.953 and 0.958 in the prototype orientation and by coordinates 0.862 to 0.867 in the IS orientation of the viral genome. However, it is not known whether these alterations are physiologically relevant to the observed changes in pathogenicity. When coinjected intraperitoneally at 50 to 100-fold excess, the nonpathogenic HSV-1 ANG protected mice against its own pathogenic variant as well as against other pathogenic HSV-1 strains. Pathogenic HSV-1 ANG proved to be genetically and phenotypically stable for at least 25 serial passages in tissue culture at either high or low multiplicity of infection. PMID- 6298474 TI - Malignant testicular germ cell tumors in father and son. A report on 2 families. PMID- 6298473 TI - Clostridial infection of a perinephric hematoma. PMID- 6298475 TI - The role of surgery following chemotherapy in stage III germ cell neoplasms. AB - After curative chemotherapy 58 patients with metastatic germinal tumors and 2 with extragonadal germinal tumors underwent an operation either to confirm a clinical complete response or to remove a residual mass. Biomarker status was converted to negative in all patients. Retroperitoneal lymphadenectomy was done in 22 patients after a complete clinical response was achieved and in 23 after an apparent complete response was achieved except for a residual retroperitoneal mass. Thoracotomy was done for a residual mass in 13 patients, while 2 underwent bilateral thoracotomy. Scarring was found in 25 patients (42 per cent), teratoma in 14 (23 per cent) and active tumor in 17 (28 per cent). One patient with scarring and primary seminoma died of disseminated seminoma, while 1 with primary teratocarcinoma died of disseminated disease, for a false negative rate of 3 per cent. Four patients with a residual mass showed unexpected findings: 2 had granulomas in the lung, 1 had carcinoid of the lung and 1 had retroperitoneal neurofibroma. Thus, there was a 7 per cent incidence of a mass being other than scar, teratoma or residual tumor. There was 1 operative death and 10 deaths of tumor, for an 18 per cent death rate. Of the 17 patients with active tumor 9 responded to salvage chemotherapy with stable complete remission and a minimum followup of 18 months. In addition to the 60 patients the advanced disease in 10 was surgically debulked after failure of chemotherapy and this procedure was followed by salvage chemotherapy. All 10 patients died, with a median survival of 7 months. The lack of responsiveness to chemotherapy was not improved by incomplete removal of tumor. PMID- 6298476 TI - Ketoconazole therapy for fungal urinary tract infections. AB - Ketoconazole was used in 11 patients to treat 12 episodes of fungal urinary infections: 6 in the upper urinary tract and 6 presumed to be in the lower urinary tract. Five patients had catheters in place. Of the 12 infections 8 were caused by Candida species, 1 by mixed Candida tropicalis and Torulopsis glabrata, and 3 by Torulopsis glabrata. Ketoconazole was administered orally at doses of 200 to 800 mg. per day for courses ranging from 5 days to more than 2 years. Five episodes of Candida urinary tract infection resolved in association with ketoconazole therapy, while only 1 of the Torulopsis glabrata infections improved. Our findings suggest that ketoconazole may be of some benefit in yeast urinary tract infections. However, the role of ketoconazole in relation to other antifungal drugs is not yet clear. PMID- 6298477 TI - delta 9-Tetrahydrocannabinol and therapeutic research legislation for cancer patients. AB - The Controlled Substances Board evaluated the implementation of the National Cancer Institute (NCI) program in Wisconsin that distributes delta 9 tetrahydrocannabinol (delta 9-THC) to cancer chemotherapy patients with nausea and vomiting refractory to conventional antiemetic drugs. The board concluded that the distribution mechanism for delta 9-THC is appropriate and adequate in Wisconsin. The drug does relieve nausea and vomiting in some cancer chemotherapy patients, but adverse side effects are prevalent. Important questions about its safety and effectiveness remain and should be resolved through scientific research and within the existing framework for testing investigational drugs that are controlled substances. "Marijuana therapeutic research" legislation, similar to that passed in 32 states, was introduced in Wisconsin after implementation and evaluation of the NCI program, but failed to recognize the existing legal framework for approving new drugs and threatened to disrupt the NCI program. With assistance from the American Cancer Society, the State Medical Society, and volunteers and professionals in cancer research, the legislation was adapted to the existing legal and administrative framework. PMID- 6298478 TI - Evaluation of hemangiomas with technetium 99m-labeled RBCs. The perfusion-blood pool mismatch. PMID- 6298479 TI - Experimental coxsackie B viral myocarditis in cynomolgus monkeys. AB - In 11 of 13 cynomolgus monkeys inoculated with coxsackievirus B3 and/or B4, myocarditis was proved histologically. Myocarditis was evident the first 10 days after inoculation and left chronic sequelae; moderate myocardial cellular hypertrophy with an increase of connective tissue with focal distribution, and some residual inflammatory foci were found 5 months after viral inoculation. Virus was recovered from the heart on the 4th day but not on the 10th day, while serum neutralizing antibody rose significantly over 2 or 3 weeks in most of the inoculated monkeys. Electrocardiographic abnormalities were found in all 11 monkeys in which the myocarditis was proved histologically. These abnormalities were usually transient and were detected most often in the ST-T segment of the right-side chest leads, corresponding to the anatomical distribution of the myocarditis. PMID- 6298480 TI - Modification of "depressed fast channel dependent slow conduction" by lidocaine and verapamil in the presence or absence of catecholamines--evidence for alteration of preferential ionic channels for slow conduction. AB - We studied, with microelectrodes, the effect of lidocaine and 1-verapamil on the upstroke phase of action potentials and conduction velocity in incompletely depolarized ventricular papillary muscle (resting potential, -58 +/- 1 mV) perfused in a high K+ (16.7 mM) Tyrode's solution. The action potential upstroke had a slur and the maximum rate of rise (Vmax) consisted of 2 components: the first large (32 +/- 7 V/sec; Vmax,fast) and the second small (10 +/- 2 V/sec, Vmax,slow). The conduction velocity was slow and ranged about 30-35 cm/sec. Isoproterenol and noradrenaline increased Vmax,slow and decreased Vmax,fast, in a concentration dependent manner (10(-8) - 10(-5)M). These effects were abolished by following application of a beta-blocker, pindolol (1 microgram/ml). Thus, the dominant ionic channel responsible for the slow conduction seemed to alter from the depressed fast channel to the slow channel as the catecholamine concentration was increased. In the absence of isoproterenol, lidocaine (2 micrograms/ml) depressed Vmax with a resultant decrease of conduction velocity to 16.4 +/- 4.2 cm/sec and which was followed by a conduction block, while 1-verapamil was without effect. In the presence of isoproterenol (5 x 10(-7) M), 1-verapamil (1 microgram/ml) depressed the Vmax and decreased the conduction velocity to 16.9 +/ 4.2 cm/sec, just before development of the conduction block, while lidocaine was without effect. These results suggest that the dominant ionic channel responsible for the slow conduction in high K media (16.7 mM), can be readily altered by changes in extracellular catecholamine concentrations and that the slowest possible conduction velocity was approximately the same (about 16 cm/sec) between the depressed fast channel-dependent and the slow channel-dependent conductions. Clinical implications of these findings were discussed. PMID- 6298481 TI - Malignant fibrous histiocytoma of the heart. AB - A case of cardiac sarcoma was reported. The tumor, which arose from the posterior wall of the left atrium, was demonstrated by echocardiography, contrast-enhanced computed tomography and direct observation during surgery. The tumor was successfully excised under emergency operation and was diagnosed as malignant fibrous histiocytoma after histologic examination. The patient had been free of symptoms for several months, but died of rapidly progressive congestive heart failure 6 months after surgery. PMID- 6298482 TI - Estimated affinity of isoproterenol to cardiac chronotropic beta-receptor and of phenylephrine to vasoconstrictive alpha-receptor of the systemic resistance vessels in human borderline hypertension. AB - There has been yet no report, to our knowledge, which clarified the affinity ("sensitivity") of catecholamines to cardiac chronotropic beta-adrenergic receptor or to vasoconstrictive alpha-adrenergic receptor of skeletal muscle resistance vessels in human borderline hypertension (BHT), using an analysis of the kinetics of the drug-receptor interaction. In the first half of this study, isoproterenol (ISO) infusion test was performed on "normal" subjects (n = 12) and subjects with BHT (n = 10). Dose-response relation was obtained in terms of various doses of ISO and the increments of heart rate (deltaHR) produced by ISO. In the latter half of this study, phenylephrine (PHE) infusion test was carried out in the series of "normal" subjects (n = 5) and subjects with BHT (n = 6), a partly different series from the first series of subjects. Dose-response relation, again, was obtained in terms of various doses of PHE and the increments of total peripheral resistance (deltaTPR) produced by PHE. These relations were displayed on Lineweaver-Burk's plot and the "affinity" of ISO to cardiac chronotropic beta-receptor or that of PHE to vasoconstrictive alpha-receptor of skeletal muscle resistance vessels was estimated, graphically, assuming that 1) drugs were diluted by a volume of body fluid equal to the blood volume which was actually measured in each individual and 2) this concentration of drugs existed around the receptor site, i.e., there was no significant individual variations in terms of the metabolism and the distribution of the drugs. Estimated "affinity" of ISO to cardiac chronotropic beta-receptor was 0.168 +/- 0.014 x 10(4) ml X ug 1 (mean +/- SE, n = 10) in patients with BHT, while it was 0.170 +/- 0.015 x 10(4) (n = 12) in "normal" subjects. Estimated "affinity" of PHE to vasoconstrictive alpha-receptor of skeletal muscle resistance vessels was 0.171 +/- 0.011 x 10(2) ml X ug-1 (n = 6) in patients with BHT, while it was 0.183 +/- 0.012 x 10(2) (n = 5) in "normal" subjects. It is concluded from these data, almost fairly, that there is no difference in terms of the estimated "affinity" of ISO to cardiac chronotropic beta-receptor or that of PHE to vasoconstrictive alpha-receptor between "normal" subjects and patients with BHT. It was also found, in PHE infusion tests, that the plasma adrenaline concentration was significantly higher in patients with BHT than in "normal" subjects, at least at two points of time in the test: a) immediately before PHE infusion, when every individual appeared to be considerably restful and b) at the end of 1-2 min after the completion of the infusion of the largest dose of PHE used. PMID- 6298483 TI - Effects of dibutyryl cyclic AMP on patients with severe heart failure. AB - Dibutyryl cyclic AMP was administered to 2 patients with severe congestive heart failure. After an intravenous drip infusion of dibutyryl cyclic AMP (300 mg/40 min), marked diuresis and mild improvement of the patients' physical activity occurred. The cardiac index increased and both systemic vascular resistance and pulmonary capillary wedge pressure decreased. Blood glucose and plasma insulin increased simultaneously, while plasma free fatty acid decreased during the infusion. A continuous intravenous micro-infusion of dibutyryl cyclic AMP (300 mg/24 h) also exerted diuresis and improved physical activity. The hemodynamic and metabolic effects of dibutyryl cyclic AMP are expected to be beneficial to a failing heart. PMID- 6298484 TI - [Recent progress in the studies of vitamin D metabolism]. PMID- 6298485 TI - [Progress in technics for analysis of the blood vitamin D3 levels]. PMID- 6298486 TI - [Biological actions and action mechanisms of vitamin D3 and its active metabolites]. PMID- 6298487 TI - [Idiopathic hypocalciuric hypercalcemia (FHH)]. PMID- 6298488 TI - [Two cases of islet cell tumor]. PMID- 6298489 TI - [Hepatocellular carcinoma with bone metastasis]. PMID- 6298490 TI - [Case of cholangiocarcinoma with acute hepatic failure]. PMID- 6298491 TI - Pseudohomozygous type II hyperlipoproteinemia. PMID- 6298492 TI - Serological survey of laboratory rodents for infection with Sendai virus, mouse hepatitis virus, reovirus type 3 and mouse adenovirus. PMID- 6298493 TI - [Effect of an oral angiotensin I-converting enzyme inhibitor on blood pressure and renin-angiotensin-aldosterone system before and after nephrectomy in two cases with acute renovascular hypertension]. PMID- 6298494 TI - Lack of active site for kyotorphin in rat mast cells: assessment by its effect on IgE-mediated 14C-serotonin release. PMID- 6298495 TI - A sero-epizootiological survey on rotavirus infection in foals. PMID- 6298496 TI - Preliminary attempts to eradicate infection with bovine leukemia virus from a stock farm in Japan. PMID- 6298497 TI - Flow-correlated influx of K, Ca, P, and Mg during continuous microperfusion of the loop of Henle in the rat. AB - Investigations on hydropenic rats were undertaken enabling us to determine the relative loop influx rates for potassium, calcium, magnesium, and phosphorus during end proximal continuous microperfusions at 15 or 35 nl . min-1 with saline free of these ions. Replicate quantitative collections were taken at collected flow rates of 9 or 26 nl . min-1. Element concentrations were determined by electron probe analysis. Results showed a significant influx of all four ions in the fluid collected at the early distal site, with magnesium showing the smallest entry rate. As expected, calcium and potassium concentrations at the early distal site increased with the perfusion flow rate. In contrast, phosphorus concentration decreased with flow. Phosphorus concentration at low flow was 0.43 +/- 0.06 mmole . liter-1 vs. 0.25 +/- 0.03 at high flow (P less than 0.01). However, increasing influx was seen with increasing flow for phosphorus (r = 0.58, P less than 0.001). Magnesium concentrations did not significantly change with flow although they were clearly different from zero: 0.12 +/- 0.02 vs. 0.10 +/- 0.02 mmole . liter-1. The rate of magnesium influx as a function of flow was also highly significant (r = 0.48, P less than 0.01). Thus, there is an increase in loop influx in all four ions with high flow rates despite disparate flow influences on early distal concentrations. In conclusion, these experiments demonstrate that (1) magnesium can enter the tubular lumen of the loop when this structure is perfused with a magnesium-free solution and (2) under our experimental conditions, phosphate can also enter the lumen suggesting that the concentration of phosphate at the early distal site is limited by the rate of entry. PMID- 6298498 TI - [Diagnosis and surgical treatment of adrenal cortical tumors]. PMID- 6298499 TI - [Current therapy of recurrent herpes]. PMID- 6298500 TI - [How does drug dependence develop?]. PMID- 6298501 TI - Morphological evidence for natural poxvirus infection in rats. PMID- 6298502 TI - A subclinical epizootic of sialodacryoadenitis in rats. AB - An epizootic of sialodacryoadenitis was identified in 10-week-old rats 5 days after they were housed with a group of older rats. The young rats generally did not develop the characteristic clinical signs of sialodacryoadenitis, but they did have typical histologic lesions of sialodacryoadenitis in Harderian, lacrimal, and salivary glands. The lesions were variable in distribution and severity, and chronic lesions persisted in the Harderian glands of the rats. PMID- 6298503 TI - Steroid structure and function--IX. Molecular conformation of catechol estrogens. PMID- 6298504 TI - Peptide-receptor protein relationships: steroid feedback in GnRH stimulation of the anterior pituitary. AB - Gonadotropin releasing hormone receptor protein (GnRH.RP) purified to homogeneity has several binding sites for its effector hormone, as evidenced from the concentration dependence binding curve. Binding of a fully biologically active iodinated GnRH decreased (P less than 0.001) by pretreatment of immobilized receptor protein with physiological concentrations of both main sex steroids, testosterone and estradiol 17-beta. With increasing concentrations of the steroids the binding of 125I-GnRH decreased biphasically for testosterone and multiphasically for estradiol 17-beta. The findings are conducive to an allosteric receptor model. PMID- 6298505 TI - The dynamics of activation of steroidogenic pathways in cultured neonatal rat ovaries. PMID- 6298506 TI - Selective chromatographic fractionation of catechol estrogens on anion exchangers in borate form. AB - The borate form of anion exchangers has been investigated for its utility in the field of estrogen analysis. The borate form of a weak (DEAE-Sephadex A-25) and a strong (QAE-Sephadex A-25) anion exchanger was easily prepared by appropriate washing of the gels, without the need of time consuming immobilization techniques. Estrogens with vicinal cis-hydroxyls were strongly retained in both gels through formation of borate complexes and readily separated from estrogens not possessing such groups. Moreover, borate complex formation with the labile o dihydroxyphenyl moiety of catechol estrogens fully protected them from decomposition during chromatography. Quantitative recovery of catechol estrogens was thereby obtained without use of antioxidants. The borate form of QAE-Sephadex A-25 was capable, in addition, of separating estrogens not possessing vicinal cis hydroxyls from the corresponding neutral steroids. PMID- 6298507 TI - Decrease of serum total and free testosterone during a low-fat high-fibre diet. AB - The concentrations of serum total and free testosterone were studied in 30 healthy, middle-aged men during a dietary intervention program. When men were transferred from their customary diet to an experimental diet, which contained less fat with a higher polyunsaturated/saturated ratio (P/S-ratio) and more fibre, there was a significant decrease in serum total testosterone concentrations (22.7 +/- 1.2 vs 19.3 +/- 1.1 nmol/l SEM, P less than 0.001). Furthermore, serum free, unbound testosterone fell from 0.23 +/- 0.01 to 0.20 +/- 0.01 nmol/l SEM (P less than 0.01). The hormonal changes were reversible. This observation suggests that testosterone activity in plasma can at least partly be modified by changing the composition of the diet. PMID- 6298509 TI - Adenosine receptors. PMID- 6298508 TI - Age-related changes in 5'-nucleotidase and alkaline phosphatase activities in mouse thymocytes. AB - Enzymatic activities of thymocytes isolated from Swiss albino mice were studied at various ages from immediately post weaning until 100 weeks of age, approaching the life expectancy of these animals. Between 5 and 10 weeks of age, the activities of lactate dehydrogenase and alkaline phosphatase decreased to a level that was maintained throughout the remainder of the aging profile. Neutral beta glycerophosphatase (pH 7.5) activity in a thymus membrane preparation was similar in all age groups. The activity of membrane-bound 5'-nucleotidase, that is, AMP hydrolyzing activity inhibited by 100 microM alpha, beta-methyleneadenosine 5' diphosphate, progressively increased as a function of age, indicating thymocyte population changes occurring very late in life. In thymocytes of the oldest mice examined (100 weeks of age), 5'-nucleotidase specific activity was approximately ten-fold greater than the activity found in 5-week-old mice. Thus, membrane-bound 5'-nucleotidase activity in thymocytes increased markedly as a function of age in Swiss albino mice; yet several other enzymatic activities, including alkaline phosphatase, remained relatively unchanged in mature mice. PMID- 6298510 TI - [Morphine-like peptides and dopaminergic pathways]. PMID- 6298511 TI - [Side effects of treatment with sulfones]. PMID- 6298512 TI - [Comparative evolution of bronchoalveolar lavage cells, angiotensin I converting enzyme, serum lysozyme, circulating immune complexes and gallium scintigraphy in mediastinopulmonary sarcoidosis]. PMID- 6298513 TI - [Importance of the administration of synacthen (beta 1-24 corticotropin) in the therapeutic protocol of patients with neoplasms in an advanced stage]. PMID- 6298514 TI - Salivary and bronchial immunoglobulins in patients with respiratory tract carcinoma. AB - Patients with carcinoma of the respiratory tract have been demonstrated to have elevated levels of immunoglobulins in the secretions bathing these areas. The clinical significance of this finding has remained unclear. In the present study, IgA and IgG levels were correlated with 5 year survival statistics in 53 patients with oral squamous cell carcinoma. Our findings indicate that there is no correlation between local IgA or IgG level and length of patient survival. PMID- 6298515 TI - Chemodectomas arising in temporal bone structures. AB - Eighteen patients with chemodectomas arising in temporal bone structures were evaluated and treated at the University of Florida. Seventeen patients have each been followed a minimum of 3 years. Patients were retrospectively staged as having "local" or "advanced" disease, depending on the presence or absence of bone destruction and/or cranial nerve involvement. Fourteen of the patients received radiation therapy as all or part of their therapy; 6 patients were treated with radiation therapy alone, 3 patients were irradiated immediately postoperatively for residual disease, and 5 patients had radiation therapy for recurrence after operation. They were treated with cobalt-60 radiation with doses ranging from 3760 to 5640 rad. All irradiated patients demonstrated evidence of tumor regression, and none have had tumor recurrence with followup of 3-12 years. Of the 8 patients with cranial nerve paralysis prior to therapy, 5 had return of function of 1 or more cranial nerves. One of 6 patients treated initially with radiation therapy had a complication, while 6 of 8 patients irradiated postoperatively had complications. None of the complications were fatal. Three patients treated by operation for early disease limited to the hypotympanum had the disease controlled for 11-12 years. Guidelines for the selection of initial therapy are discussed. PMID- 6298517 TI - Pulse-chase studies of the POMC/beta-endorphin system in the pituitary of acutely and chronically stressed rats. AB - Experiments were carried out to determine whether stress induces biochemical changes in the pro-opiomelanocortin (POMC) system in anterior (AL) and intermediate-posterior lobe (IPL) of rat. In a series of pulse-chase experiments, acute stress led to an increase in POMC biosynthesis and shorter half-life in AL. However, when the animals were chronically stressed, the AL no longer exhibited increased POMC synthesis. On the other hand, in the IPL, acute stress did not produce any biochemical changes, but chronic stress led to an increase in POMC synthesis and shorter half-life. These data suggest that AL and IPL are affected by acute and/or chronic exposure to stress in opposite directions and that the POMC system in AL may play an important role in stress-induced analgesia. PMID- 6298516 TI - Different receptors mediate morphine-induced prolactin and growth hormone release. AB - Morphine has a variety of actions on the release of pituitary hormones in addition to its analgesic actions. In the rat, morphine releases prolactin and growth hormone at doses comparable to those active in the tailflick analgesic assay. Naloxazone selectively inhibits the high affinity (mu 1) binding site and dramatically decreases morphine's analgesic potency for over 24 hours. In an effort to determine the opiate receptor mechanisms of morphine-induced prolactin and growth hormone release, groups of rats were treated with either naloxone or naloxazone and 24 hours later the prolactin and growth hormone response to morphine sulfate determined. Peak prolactin levels in the group treated the previous day with naloxone were very similar to levels in untreated controls. However, the peak prolactin levels in the group that received naloxazone the day before were depressed 80% (p less than 0.005). By contrast, growth hormone levels in the naloxazone-treated animals were actually higher than in the naloxone treated group. Thus prolactin release, like analgesia, appears to be mediated through high affinity, or mu 1, sites, while growth hormone release is not. PMID- 6298518 TI - In vivo studies with ICI 154,129, a putative delta receptor antagonist. AB - We studied the in vivo pharmacology of ICI 154,129, a new antagonist that is claimed to show selectivity for delta opiate receptors. At s.c. doses of 30 and 100 mg/kg, ICI 154,129 had no marked effect on the gastrointestinal transit of a charcoal meal in mice. In this test, ICI 154,129 reversed the inhibitory action of metkephamid (a proposed delta receptor agonist) but not that of levorphanol. ICI 154,129 was proconvulsant in the mouse picrotoxin potentiation test; the dose response curve had a low ceiling and was biphasic. Naloxone (1 mg/kg, s.c.) enhanced the proconvulsant action of ICI 154,129 (40 mg/kg, s.c.) by an unknown mechanism. PMID- 6298519 TI - Motility effects of opioid peptides in dog intestine. AB - Six opioid peptides, like morphine, were found to produce dose-dependent contractions of dog isolated intestine when administered as intraarterial boluses. The increases in incidence and amplitude of intestinal contractions were antagonized by naloxone. The rank order of potency of the opioid agonists tested was D-Ala2-met-enkephalinamide greater than D-Ala2-leu-enkephalinamide greater than met-enkephalin greater than beta-endorphin 1-31 greater than morphine greater than morphiceptin greater than dynorphin 1-13. The contractions induced by two opioid agonists displayed differential sensitivity to blockade by tetrodotoxin (TTX). Met-enkephalin was barely affected by concentrations of TTX that markedly reduced responses to morphiceptin. Some portion of the motility effect of metenkephalin may be exerted directly on intestinal smooth muscle. PMID- 6298520 TI - Postulated thermoregulatory roles for different opiate receptors in rats. PMID- 6298521 TI - Morphine-like analgesic actions of enkephalin-like peptides containing the Tyr Arg unit. AB - The analgesic actions of some synthetically prepared peptides having the Tyr-D Arg unit at the N terminal portion of met- and leu-enkephalin were measured by the intra-cisternal injection method in mice. Among them, Tyr-D-Arg-Gly-Phe (DR 4) induced the most potent naloxone-reversible analgesia and was also effective by s.c. injection. DR-4 showed the good affinity to mu-receptor, and the resistance to the enzymatic degradation. PMID- 6298522 TI - Effects of various opiates including specific delta and mu agonists on dopamine release from nigrostriatal dopaminergic neurons in vitro in the rat and in vivo in the cat. PMID- 6298523 TI - Calmodulin increases in selective brain regions with opioid dependence. AB - Acute challenge of thalamic membranes with opioid agonists displaces calcium and prevents isoproterenol stimulation of adenylate cyclase. Chronic morphine administration for three days or three weeks results in significant increases in the level of calmodulin in membranes of thalamus, but not in periaqueductal gray, striatum, amygdala or hypothalamus. PMID- 6298524 TI - Role of opioid receptors in the substantia nigra in morphine-induced muscular rigidity. AB - Uni- or bilateral injection of morphine (MO) (3-13 nmoles) into the substantia nigra pars reticulata (SNR) produced tonic activity in the electromyogram (EMG) recorded from the gastrocnemius-soleus (GS) muscle of non-anesthesized rats. This activity was antagonized by naloxone (NAL) (10 nmoles) coadministered with MO into the SNR. Bilateral lesion of the caudate nucleus (CN) with kainic acid did not prevent the tonic EMG activity occurring after the injection of MO into the SNR. Unilateral injection of MO (40 nmoles) into the CN also induced tonic EMG activity in the GS-muscle, which was antagonized by NAL (10 nmoles) administered into the SNR ipsilaterally and simultaneously to the intrastriatal injection of MO. The results suggest that enkephalinergic mechanisms in the SNR seem to play a crucial role in the function of striatal efferent pathways relayed in the SNR. PMID- 6298525 TI - Disinhibition of nigral GABA output neurons mediates muscular rigidity elicited by striatal opioid receptor stimulation. AB - Unilateral injection of bicuculline (12.5-50 ng) into the substantia nigra pars reticulata (SNR) dose-dependently produced a tonic activity in the electromyogram (EMG), which was antagonized by coadministered muscimol. Bilateral lesions of the caudate nucleus with kainic acid failed to affect the EMG activity produced by unilateral injection of bicuculline into the SNR. Administration of muscimol (12.5 and 25 ng) into the SNR antagonized the tonic activity in the EMG produced by morphine (15 mg/kg i.p.), whereas bicuculline (50 ng) enhanced it. These results suggest that an impairment in the GABAergic neurotransmission in the SNR is relevant for mediating the tonic activity in the EMG produced by morphine and that GABAergic mechanisms in the SNR play an important role in the modulating the opioid-induced alterations in the striatal function. PMID- 6298526 TI - Multiple opiate receptors on neurons of the mammalian central nervous system. In vivo and in vitro studies. AB - Experiments were performed in rat spinal cord cells in vivo and on hippocampal pyramidal cells in vitro. These investigations suggest that acute and chronic treatment renders the neurons subsensitive to opiate alkaloids without altering their sensitivity to opioid peptides. The experiments performed in the dorsal horn of the spinal cord provide evidence that in this structure mu- and delta receptors may also be localized on the same cell. The evidence for the existence of distinct types of opiate receptors as originally proposed by (1) and suggested by the differing pattern of opiate and opioid peptide activity in various assay systems has been substantiated by investigations involving the selective development of tolerance and the protection of a particular receptor subtype by chemical manipulation. Furthermore, they have been characterized by the use of low concentrations of radiolabelled agonists and antagonists and through the ability of GTP to influence differentially their binding to the opiate receptor (for refs. see: 2). Recently autoradiographic techniques were able to provide direct evidence by mu- and delta-receptors in the mammalian brain (3; 4; 5; 6; and cits. therein). The presence of multiple opiate receptors located on the same cell is suggested by the present study. PMID- 6298527 TI - Estimation in vivo of the receptor constants of morphine in naive and morphine tolerant rats. AB - The efficacy and dissociation constant of morphine in naive and morphine-tolerant rats were estimated by the method of partial irreversible blockade of a fraction of the receptor population with buprenorphine. The dissociation constant (KA) of morphine increased from 3.3 x 10(-5) M in naive to 1.4 x 10(-4) M in morphine tolerant animals, indicating a decrease in the affinity of morphine for its receptor in the tolerant state. The efficacy of morphine (KA/A50 + 1) was constant in naive and tolerant animals (4.23 and 4.46, respectively). When the data were recalculated following conversion of administered dose to brain morphine concentration, the value of KA was 1.7 x 10(-7) M in naive and 7.7 x 10( 7) M in morphine-tolerant rats, while the efficacy was 2.5 and 3.4, respectively. In addition, the stimulus-effect relationship varied in the two states, with the curve in the tolerant animal being of different shape and broader range than in the naive rat. The present results suggest that (a) tolerance to opiate agonists may involve affinity changes and (b) post-receptor events leading to the measured effect may also be affected. PMID- 6298528 TI - Physiological analysis of delta opioid receptors in the guinea pig myenteric plexus. AB - Ilea taken from guinea pigs that had been chronically exposed to morphine exhibit a greater tolerance to morphine and normorphine than to the opioid peptides D ala2-D-leu5-enkephalin (DADLE) or D-met2-pro5-enkephalinamide (DMPE). This differential tolerance strongly implies the existence of at least two different types of opioid receptor in the guinea pig myenteric plexus or two different mechanisms of interaction between opioids and their receptor complex. Since DADLE is considered to be the prototypic ligand for the delta receptor, the above results imply the presence of delta receptors in the guinea pig myenteric plexus and furthermore, that this subtype of opioid receptor is associated with the modulation of release of enteric acetylcholine. PMID- 6298530 TI - Stimulation of beta-endorphin and beta-lipotropin release from the anterior but not the neurointermediate pituitary lobe in the rat after acute administration of serotonin-acting drugs. PMID- 6298529 TI - Compounds of novel structure having kappa-agonist behavioral effects in rhesus monkeys. AB - The kappa-agonist behavioral effects of several compounds were studied in rhesus monkeys and mice. Rhesus monkeys trained to discriminate ethylketazocine from saline responded as if ethylketazocine had been administered when given bridged oripavines with either N-allyl or N-cyclopropylmethyl, but not N-methyl, substituents. These compounds had C7 substitutions of either 2-hydroxy-2-pentyl or 2-hydroxy-5-methyl-2-hexyl. Monkeys also showed ethylketazocine-like responding when given U-50,488 (trans-3,4-dichloro-N-methyl-N-[2- (1 pyrrolidinyl) cyclohexyl]-benzeneacetamide), a compound with an atypical structure not resembling any known narcotic. Additionally, ethylketazocine-like responding was produced by two 5,9-alpha dimethyl 6-7-benzomorphans with either an N-2-methoxyisobutyl or an N-2-methoxy-propyl substituent. The latter compound was the only compound active in producing ethylketazocine-like discriminative effects that also reversed morphine-withdrawal signs. The N-methyl bridged oripavines that were inactive in producing ethylketazocine-like discriminative effects reversed morphine withdrawal signs. PMID- 6298531 TI - Alkylxanthines as adenosine receptor antagonists and membrane phosphodiesterase inhibitors in central nervous tissue: evaluation of structure-activity relationships. PMID- 6298532 TI - Further evidence for the involvement of D2, but not D1 dopamine receptors in dopaminergic control of striatal cholinergic transmission. PMID- 6298533 TI - Alpha 2 adrenergic receptors are located prejunctionally in the Auerbach's plexus of the guinea pig small intestine: direct demonstration by radioligand binding. PMID- 6298534 TI - Naloxone potentiation of apomorphine-induced stereotypic climbing in mice and interaction with mu-, sigma- and kappa-opiate drugs. PMID- 6298535 TI - Differential recognition of "enkephalinase" and angiotensin-converting enzyme by new carboxyalkyl inhibitors. AB - New carboxylalkyl compounds derived from Phe-Leu and corresponding to the general formula C6H5-CH2-CH(R)CO-L.Leu with R = -COOH, 3, R = -CH2-COOH, 4, R = -NH-CH2 COOH, 5, R = -NH-(CH2)2-COOH, 6, have been found to inhibit the breakdown of the Gly3-Phe4 bond of [3H] Leu-enkephalin or [3H]D.Ala2-Leu-enkephalin resulting from the action of the mouse striatal metallopeptidases: "enkephalinase" or angiotensin-converting enzyme (A.C.E.). The carboxyl coordinating ability of the Zn atom seems to be significantly higher in ACE than in "enkephalinase". Moreover, IC50 values against "enkephalinase" were found in the same range whatever the length of the chain bearing the carboxyl group whereas a well defined position of this group with respect to the Zn atom is required for strong ACE inhibition. These features suggest a larger degree of freedom of the carboxyalkyl moieties within the active site of "enkephalinase". Therefore the differential recognition of active sites of both peptidases leads to: i) N (carboxymethyl)-L-Phe-L-Leu, 5, a competitive inhibitor of "enkephalinase" (KI = 0.7 microM) and ACE (KI = 1.2 microM) which could be used as mixed inhibitor for both enzymes; ii) N-[(R,S)-2-carboxy, 3-benzylpropanoyl]-L-Leucine, 3, a full competitive inhibitor of "enkephalinase" (KI = 0.34 microM) which does not interact with ACE (IC50 greater than 10,000 microM). This compound can be considered as the first example of a new series of highly potent and specific "enkephalinase" inhibitors. PMID- 6298536 TI - Responses mediated via beta 1, but not beta 2-adrenoceptors, exhibit hypothermia induced supersensitivity. AB - The beta-adrenoceptor-mediated responses of various isolated tissues from the guinea-pig were examined at bath temperatures of 38 and 30 degrees C. The positive inotropic responses to orciprenaline of paced left atria and papillary muscles were potentiated at the lower bath temperature, as indicated by a significant shift of the dose-response curve to the left. A similar hypothermia induced supersensitivity was observed for the positive chronotropic response of right atria. The beta-adrenoceptor-mediated inhibition of coaxially stimulated ileum in response to orciprenaline also exhibited supersensitivity at 30 degrees C. In contrast, the relaxation of lung strips was not altered at the lower bath temperature. The relaxant responses of potassium-contracted uterine strips from untreated guinea-pigs or progesterone-dominated guinea-pigs and rats also failed to reveal any hypothermia-induced supersensitivity. The responses of lung and uterine strips are mediated via beta-adrenoceptors of the beta 2-type, whereas they are of the beta 1-type in the cardiac preparations and ileum. Therefore, these results suggest that hypothermia-induced supersensitivity occurs only at the beta 1-adrenoceptors, indicating a fundamental temperature-dependent difference between the two receptor types. PMID- 6298537 TI - Stereochemical requirements of alpha 2-adrenergic receptors for alpha-methyl substituted phenethylamines. AB - The alpha 1- and alpha 2-adrenergic effects of the stereoisomers of alpha methyldopamine were evaluated in guinea pig aorta and field-stimulated guinea pig ileum, respectively, in order to establish the stereochemical requirements of these receptors for alpha-methyl substituted phenethylamines. The alpha 1 adrenergic receptor did not distinguish between the stereoisomers of alpha methyldopamine which is in marked contrast to the alpha 2-adrenergic receptor where a dramatic stereochemical preference for the 2S(+)-isomer was observed. In addition, 2R(-)-alpha-methyldopamine displayed no alpha-receptor subtype specificity whereas 2S(+)-alpha-methyldopamine was highly selective (23 fold) for the alpha 2-adrenergic receptor. These results indicate that the alpha 2 adrenergic receptor can recognize and accept methyl substituents at the alpha carbon atom of phenethylamines when correctly oriented, while the alpha 1 adrenergic receptor cannot. Thus, the alpha-carbon atom is a major determinant of the alpha 2-adrenergic effects of phenethylamines, and plays an important role in determining alpha-receptor subtype specificity. It is hypothesized that the alpha 2-adrenergic receptor (but not alpha 1) has an additional recognition site which will accommodate alpha-substituted phenethylamines. PMID- 6298538 TI - Processing of normal and non-glycosylated forms of toad pro-opiocortin by rat intermediate (pituitary) lobe pro-opiocortin converting enzyme activity. AB - The influence of glycosylation of a prohormone, pro-opiocortin, on its processing by intermediate (pituitary) lobe converting enzyme activity in vitro was studied. [3H]-arginine-labeled glycosylated and non-glycosylated pro-opiocortins were isolated from untreated, and tunicamycin treated toad neurointermediate lobes, respectively, after pulse-labeling in [3H]-arginine containing incubation media. These labeled precursors were then incubated at 37 degrees C in the presence of pro-opiocortin converting enzyme activity derived from rat intermediate lobe (pituitary) secretory granule lysates. The rates of conversion of the glycosylated and nonglycosylated pro-opiocortins to smaller peptide products, in vitro, were similar. Analysis of the peptide products by immunoprecipitation with ACTH and beta-endorphin antisera, and subsequent electrophoresis on acid-urea gels, indicate a comparable processing in vitro of the two forms of pro opiocortin substrate. The only difference was that the normally glycosylated peptide products derived from glycosylated pro-opiocortin (i.e., 13K ACTH, 21K ACTH, and the 16K glycopeptide) differed in their gel electrophoretic mobilities from their counterparts derived from nonglycosylated prohormone, in a manner consistent with the absence of carbohydrate on the latter's peptides. These data show that glycosylation of the prohormone does not influence its processing in vitro by the converting enzyme activity. PMID- 6298539 TI - Site of calcium requirement for stimulation of ACTH release in rat anterior pituitary cells in culture by synthetic ovine corticotropin-releasing factor. AB - Synthetic ovine corticotropin-releasing factor (CRF) causes a 6- to 8-fold stimulation of ACTH release and cAMP accumulation in rat anterior pituitary cells in culture at ED50 values of 1 and 4 nM, respectively. Removal of Ca2+ from the incubation medium reduces CRF-induced ACTH release by 70% but have no effect on cyclic AMP accumulation. ACTH release induced by 8-Br-cAMP is inhibited by 65% in the absence of Ca2+. The Ca2+ ionophore A23187 does not alter spontaneous ACTH release. Verapamil, a pharmacological agent that blocks Ca2+ entry into cells, has no influence on spontaneous or CRF-induced ACTH release. The present data clearly demonstrate a role of Ca2+ in CRF action at a step subsequent to cAMP formation and suggest that Ca2+ is mobilized from intracellular stores during CRF stimulation. PMID- 6298540 TI - Multicomponent structure of the thyrotropin receptor: relationship to Graves' disease. AB - The thyrotropin receptor is proposed to contain both a glycoprotein and a ganglioside component. Monoclonal antibodies have been developed against soluble thyroid TSH receptor preparations and using Graves' lymphocytes. These show that initial recognition of thyrotropin requires the glycoprotein component, but that monoclonal antibodies to this component block thyrotropin function (blocking antibodies) rather than mimic thyrotropin. Monoclonal antibodies which stimulate thyroid activity in cultured cell systems (cAMP increase) or mouse bioassays, all interact with gangliosides. Using monoclonal antibodies to the glycoprotein component of the thyrotropin receptor, we show that two protein bands, molecular weights 18,000-23,000 and 50,000-55,000, are precipitated from detergent solubilized preparations. Using a crosslinking procedure with 125I-labeled thyrotropin, we show that thyrotropin binding is related to the disappearance of the 18,000-23,000 molecular weight band on sodium dodecylsulfate gels and the appearance of a 30,000-33,000 molecular weight thyrotropin-membrane component complex. Higher molecular weight thyrotropin-membrane complexes of 75,000-80,000 and 250,000 are visualized when binding studies are performed at pH 7.4 in physiologic medium; larger amounts of the 30,000-33,000 complex are evident at pH 6.0 in a low salt medium. It is thus proposed that the glycoprotein component of the thyrotropin receptor is composed of two subunits with apparent molecular weights of 18,000-23,000 and 50,000-55,000; that the 18,000-23,000 subunit interacts with thyrotropin; and that different receptor subunits can exist depending on in vitro binding conditions. Using monoclonal-stimulating antibodies or natural autoimmune IgG preparations from patients' sera, we show that stimulating antibodies exhibit species-specific binding to human thyroid ganglioside preparations. Individual components or determinants of the thyrotropin receptor structure with specific autoimmune immunoglobulins. PMID- 6298541 TI - Stimulation of iodine organification in porcine thyroid cells by thyroid stimulators. AB - Several Graves' sera were simultaneously assessed in a bioassay based on the ability of porcine thyroid cells to organify 125I and in a radioreceptor assay for TSH receptor binding activity. Both assay systems were sensitive to 1 mcU/ml (final concentration) of unlabelled bovine TSH. Six Graves' sera were studied in detail over a wide (0-1.0 mcl sera) dose response range in repeat determinations. Two sera exhibited parallel binding and stimulating. However, two sera revealed significant inhibition of 125I-TSH binding prior to the demonstration of stimulation and the other two sera showed stimulatory capabilities before significant binding was evident. IgG was prepared from one serum by ammonium sulphate precipitation and chromatography on Sepharose 6B and then subjected to preparative isoelectric focusing. The isoelectric distribution of the two activities were found to be identical with major peaks of activity at pl=9.5 and pl=8.5. In summary: 1) each Graves' sera exhibits different dose-response curves with respect to binding and stimulation, 2) at certain concentrations of sera, only binding or stimulation were evident, 3) neither assay was consistently more sensitive for the presence of Graves' immunoglobulins, 4) for one Graves' sera, binding and stimulation could not be separated by isoelectric focusing. These studies would suggest each Graves' immunoglobulin has inherently different characteristics in its interaction with the TSH receptor. PMID- 6298543 TI - Chronic caffeine consumption increases the number of brain adenosine receptors. AB - Caffeine, a potent central stimulant, is known to competitively inhibit the specific binding of both adenosine and benzodiazepine receptor ligands to brain membranes in vitro. In mice receiving a diet containing non-toxic doses of caffeine (200 or 400 mg/kg diet) for periods up to 40 days, a dose-related increase in the number of binding sites for [3H]-CHA and [3H] DPX was observed in whole brain membranes without modifications of the receptors' affinity. Furthermore, a transitory increase in the number of [3H]-DZP binding sites was observed. These preliminary data seem to confirm the involvement of the adenosine receptors in the mode of action of caffeine and may be relevant to the development of both tolerance and dependence to some of the central effects of this compound. PMID- 6298542 TI - Interactions between phencyclidine and delta 9-tetrahydrocannabinol in mice following smoke exposure. AB - The behavioral and pharmacological interactions between delta 9 tetrahydrocannabinol (delta 9-THC) and phencyclidine (PCP) were studied following coadministration of the drugs in smoke to mice. While delta 9-THC (25, 50 or 100 mg/cigarette) had little effect on spontaneous motor activity, all doses attenuated the hyperactivity elicited by PCP X HCl (25 and 50 mg/cigarette). delta 9-THC produced a dose-related hypothermia. PCP X HCl (50 mg/cigarette) had no effect on body temperature but enhanced hypothermia when combined with 25 mg of delta 9-THC. delta 9-THC (100 mg/cigarette) had no effect on the biodisposition of 3H-PCP and its pyrolytic product, 3H-phenylcyclohexene (3H-PC), when examined immediately after 3H-PCP X HCl (50 mg/ cigarette) exposure. At 30 min, brain, liver, lung and plasma contained higher concentrations of 3H-PC and fat and plasma contained lower concentrations of 3H-PCP in the mice exposed to both drugs compared to 3H-PCP X HCl alone. It appears, therefore, that delta 9 THC has the potential for altering the behavioral, pharmacological and pharmacokinetic sequelae of PCP abuse. PMID- 6298544 TI - In vivo lactogenic effects of anti prolactin receptor antibodies in pseudopregnant rabbits. AB - Antibodies generated against partially purified prolactin receptors from rabbit mammary gland membranes were tested for their effects on prolactin binding to receptors and for their in vivo biological potencies. These antibodies are able to inhibit prolactin binding to crude rabbit mammary gland membranes. When administered intravenously or intramuscularly to pseudopregnant rabbits, they induce respectively an accumulation of beta-casein or an enhancement of beta casein synthesis and mRNA concentration in the mammary gland. Moreover the stimulatory effect of these anti-prolactin receptor antibodies on casein synthesis is totally abolished by a simultaneous treatment with progesterone, which is a potent in vivo inhibitor of prolactin action. These results better establish the prolactin-like activities of these antibodies previously observed in vitro and give strong support to the hypothesis that prolactin molecule is not required beyond the initial binding to its receptor to induce hormonal effects. PMID- 6298545 TI - 60-Hz electric field alters the steroidogenic response of rat adrenal tissue, in vitro. AB - Exposure to a 60-Hz electric field at 10 kV/m but not at 5 kV/m, 100 kV/m or 1000 kV/m caused a highly significant, threefold elevation in the steroidogenic response of rat adrenal cortical tissue after the administration of 10 mU of adrenocorticotrophic hormone (ACTH) under in vitro, superfusion conditions. A 60 Hz electric field can directly influence the function of mammalian tissue in the absence of central-nervous-system mediation. PMID- 6298546 TI - Enhanced hepatotoxicity and inhibition of liver endoplasmic reticulum calcium pump by CCl4 in rats fed a thiamine deficient diet. AB - Male Sprague-Dawley rats were fed a thiamine deficient diet for three weeks, then treated with a range of CCl4 doses (0.01-1-ml/kg). Rats fed the deficient diet grew more slowly (body weight 65 percent of control) and had elevated liver glutathione (GSH) (220 percent of control). CCl4 hepatotoxicity, assessed by serum glutamicpyruvic transaminase (SGPT) activity and histological examination 24 hours after the hepatotoxin, was augmented in the group fed the thiamine deficient diet. Likewise, CCl4 inhibition of liver endoplasmic reticulum (ER) function (glucose-6-phosphatase (G6Pase) and calcium pump activities one hour after CCl4) was enhanced in rats fed the deficient diet. These results suggest that thiamine deficiency enhances CCl4 damage to membranes of the ER and enhances CCl4 hepatotoxicity. PMID- 6298547 TI - Changes in erythrocyte sodium, sodium transport and 3H ouabain binding capacity during digoxin administration in the pig. AB - Time course of the changes in erythrocyte sodium content, sodium transport, 3H ouabain binding capacity and Na+, K+-ATPase activity were measured for 14 weeks, in 6 young pigs treated with digoxin and in 6 control pigs. After one week of treatment the erythrocyte sodium content increased from 5.4 mmol/kg cells to 6.9 mmol/kg cells and the efflux rate constant of sodium decreased. With prolonged treatment the erythrocyte sodium content returned to normal and the 3H ouabain binding capacity increased by week 5. The plasma digoxin concentration decreased from 1.1 ng/ml at week 5 to 0.6 ng/ml at week 8 probably due to the decline in dose (microgram/kg) of digoxin with age. The efflux rate constant of sodium and Na+, K+-ATPase activity were higher towards the end of treatment. It is concluded that with prolonged administration of digoxin there is an increase in erythrocyte sodium pump units. PMID- 6298548 TI - The role of calmodulin in insulin secretion: the presence of a calmodulin stimulatable phosphodiesterase in pancreatic islets of normal and pregnant rats. PMID- 6298549 TI - Antagonistic effects of naloxone on CCK-octapeptide induced satiety and rumino reticular hypomotility in sheep. AB - Continuous intracerebroventricular (ICV) infusion of CCK-octapeptide (CCK8) was performed in ewes fitted with a permanent cannula into the lateral cerebral ventricle and Nichrome electrodes on the reticulum in order to record its electrical activity. In the first series of experiments, subsequently repeated in 12 h fasted animals, CCK8 was infused during the first hour of a 3 hour period of feeding at 2.5, 5 and 10 ng.kg-1.min-1. The same series of infusion were performed 20 min after ICV injection of 2.4 and 10 micrograms.kg-1 of naloxone. CCK8 reduced significantly in a dose related manner the food intake (r = 0.95; P less than 0.01) and the frequency of cyclic spike bursts associated to biphasic contractions of the reticulum observed during feeding (r = 0.89; P less than 0.01). At 5 and 10 ng.kg-1.min-1, the reduction of food intake reached 46.2 and 52.6% during the period of infusion; the basal and stimulated (feeding) frequency of reticular contractions were nearly halved. Previous ICV administration of naloxone (2.4 micrograms.kg-1) partially blocked the effects of CCK8 infusion on both food intake (72%) and reticular frequency (54% basal, 67% stimulated). The CCK8 induced effects on both food intake and frequency of reticular contraction were completely abolished after a previous 10 micrograms.kg-1 injection of naloxone. These results suggest that the central effects of CCK8 on feeding behavior and forestomach motility involve similar central structures and are mediated through opiate receptor structures. PMID- 6298550 TI - Increase in brain 125I-cholecystokinin (CCK) receptor binding following chronic haloperidol treatment, intracisternal 6-hydroxydopamine or ventral tegmental lesions. AB - Specific 125I-CCK receptor binding was significantly increased in brain tissue taken from guinea pig or mouse following chronic (2-3 week) daily administration of haloperidol (2-3 mg/kg/day). Scatchard analysis indicated the increase in CCK binding was due to an increased receptor number (B max) with no change in affinity (Kd). In guinea pigs, the increased CCK binding was observed in the mesolimbic regions and frontal cortex, but not in striatum, hippocampus nor posterior cortex. In mice, however, the increases occurred in both pooled cerebral cortical-hippocampal tissue, and in the remainder of the brain. Enhanced CCK receptor binding was also observed in membranes prepared from whole brain of mice one month following intracisternal injection of 6-hydroxydopamine. Additionally, an increase in CCK binding was observed in mesolimbic regions and frontal cortex, but not striatum or hippocampus, of guinea pigs 3 weeks after an unilateral radiofrequency lesions of the ipsilateral ventral tegmentum. The present studies demonstrate that three different procedures which reduce dopaminergic function in the brain enhance CCK receptor binding. The data provide further support for a functional interrelationship between dopaminergic systems and CCK in some brain regions and raise the possibility that CCK may play a role in the antipsychotic action of neuroleptics. PMID- 6298552 TI - CL 218872 antagonism of diazepam induced loss of righting reflex: evidence for partial agonistic activity at the benzodiazepine receptor. AB - A recent hypothesis suggests that the "selective anxiolytic" activity of the triazolopyridazine, CL 218872, is a reflection of this compounds high affinity for a benzodiazepine (BZD) receptor subtype. Subsequent to this proposal, the observation was made that CL 218872 does not effectively discriminate BZD receptor subtypes in vitro at physiological temperatures (37 degrees C). Based upon this observation, a selective effect in vivo related to the high affinity of CL 218872 for a BZD receptor subtype appears unlikely. The present study provides evidence for an alternative hypothesis to explain the unique pharmacological properties of CL 218872. The ability of CL 218872 to antagonize diazepam induced loss of righting reflex and enhance the anticonvulsant effect of diazepam in mice suggests that this triazolopyridazine may act as a partial agonist at the BZD receptor. Compared to the pharmacologically active BZDs, the unique actions of CL 218872 may be related to the lower intrinsic activity of this compound. PMID- 6298551 TI - A role for the lipoxygenase pathway of arachidonic acid metabolism in glucose- and glucagon-induced insulin secretion. AB - Although the cyclo-oxygenase pathway of arachidonic acid (AA) metabolism inhibits glucose-stimulated insulin release through synthesis of prostaglandins, very little attention has been given to the effects of lipoxygenase pathway products on beta cell function. We have examined the effects of two structurally dissimilar lipoxygenase inhibitors on insulin release from monolayer-cultured rat islet cells. Both nordihydroguaiaretic acid (NDGA, 20-50 microM) and BW755c (100 250 microM) caused a dose-responsive inhibition of glucose-induced insulin release. This inhibitory effect occurred despite concomitant inhibition of prostaglandin E synthesis. Lipoxygenase inhibitors also impeded cyclic AMP accumulation. Insulin and cyclic AMP release induced by glucagon were also blunted. These studies suggest the hypothesis that AA released in or near the beta cell is metabolized to lipoxygenase product(s) which have feed-forward properties important to glucose- and glucagon-stimulated cyclic nucleotide accumulation and insulin release. PMID- 6298553 TI - Regulation of sodium and water excretion by catecholamines. AB - There is considerable evidence that the renal nerves contribute to the regulation of salt and water excretion by a direct effect on tubular reabsorption, independent of changes in renal hemodynamics. Whereas the effect of the adrenergic nervous system on sodium reabsorption appears to be established in anesthetized animals, it has been suggested that the basal activity of the renal sympathetic nerves in conscious dogs is too low to have a significant effect on sodium reabsorption by the proximal tubules. However, denervation natriuresis and diuresis have recently been demonstrated in conscious euvolemic and conscious volume-expanded rats. The effects of renal nerve stimulation on the handling of sodium and water by the proximal tubule can be mimicked by infusion of the alpha adrenergic agonist norepinephrine and prevented by infusion of an alpha adrenergic antagonist. This confirms that they are mediated by alpha-receptors. The adrenergic nervous system may have an independent role in the control of sodium excretion or may be complementary to other systems such as the renin angiotensin-aldosterone system. PMID- 6298555 TI - [Infiltrative-nodular form of neoplasm metastasis to the pleura]. PMID- 6298554 TI - Modulation of tissue prostaglandin synthesizing capacity by increased ratios of dietary alpha-linolenic acid to linoleic acid. AB - Semipurified diets containing ratios of alpha-linolenic acid (18:3 omega 3) to linoleic acid (18:2 omega 6) of 1/32, 1/7, 1/1, and 3.5/1 in the form of corn oil, soybean oil, soybean/linseed oil mix and linseed oil were fed to rats for 2 months. The first 3 diets were fed to another group of rats for 4 months and to a group through the second generation. Fatty acid analysis of liver and spleen ethanolamine glycerophosphatide revealed that, as the level of 18:3 omega 3 in the diet increased, the elongated, desaturated metabolites of the omega 6 series decreased and the omega 3 series increased. Noteworthy was the depression in the amount of the precursor of the 2-series prostaglandins (PG) as the omega 3 levels increased. Synthesis of PG by liver of rats fed 2 or 4 months markedly decreased, but at 2 months in thymus and spleen, it showed a trend toward decreasing only. Brain slices showed no decrease in PGF2 alpha synthesis after 4 months, but did decrease significantly after feeding the diets to the second generation. Synthesis of PGE2 by spleen homogenate from the second generation also significantly decreased. The replacement of omega 6 series fatty acids by omega 3 series is explained by the effective competition of 18:3 omega 3 over 18:2 omega 6 for the delta 6 desaturase. Depressions in PG synthesis by high dietary 18:3 omega 3 is explained by the competitive inhibition of the PG synthetase complex by 20:5 omega 3 as well as by the decreased levels of 20:4 omega 6. PMID- 6298556 TI - [Metastasis of hypernephroid carcinoma to the bones detected by 99mTc dimercaptosuccinic acid]. PMID- 6298557 TI - [Scintigraphy of the parasternal lymph nodes in breast cancer]. PMID- 6298558 TI - [Intravenous metopiron test in normal persons and patients with adrenal cortex diseases]. PMID- 6298559 TI - [Lambert-Eaton syndrome. A contribution to its pathogenesis, diagnosis and therapy]. PMID- 6298560 TI - [Poisoning danger in high dosage vitamin D therapy of dermatologic diseases]. PMID- 6298561 TI - [Hepatitis A in homosexual males]. PMID- 6298562 TI - Herpes zoster-associated encephalitis: clinicopathologic report of 12 cases and review of the literature. AB - Herpes-zoster associated encephalitis (HZAE) is an uncommon complication of herpes zoster. Over 8 years, we evaluated 12 patients with this clinical diagnosis. The majority of our patients were elderly, immunosuppressed, and found to have disseminated skin lesions prior to the onset of CNS symptoms. All patients had abnormal EEGs, and CSF pleocytosis was found in most. In the seven patients who were tested, specific antibody to the varicella-zoster membrane antigen (FAMA) was detected in spinal fluid during the course of the illness. Although three patients died during the period of active infection, the virus could not be definitively implicated as the cause of death. These HZAE patients could not be distinguished from our other herpes zoster patients on the basis of age, initially involved dermatome, or mortality rate. However, among herpes zoster patients who survived, duration of hospitalization was significantly longer in those with a diagnosis of HZAE. All surviving HZAE patients had a slow but eventual return to their prior cognitive status. PMID- 6298563 TI - The influence of dust originated in different coal mines on pneumoconiotic changes in white rats. AB - In experiments on white rats, three different types of dust of equal dispersion, coming from coal mines, were examined. The dusts exhibited different content of free silica and certain metals, such as: nickel, chromium and vanadium. The animals intratracheally administered with 50 mg of the mentioned dusts had hydroxyproline determined. After six months of experiment, histopathological examination of lungs was carried out. It was found that different SiO2 content (5 and 11%) with the same content of trace metals and different content of trace metals with the same SiO2 content (5%) does not affect hydroxyproline content in lungs. A significant increase in hydroxyproline level in rats' lungs was found in a group that obtained dust of the highest SiO2 content (11%) with more trace metals. Findings of histopathological examinations confirmed the data obtained from hydroxyproline levels measurements. It seems that compounds of certain metals, such as: nickel, chromium, vanadium, at a specific SiO2 level in the dust may increase its fibrogenic properties in lungs. PMID- 6298564 TI - [Liver enzymes in the guinea pig after chronic oral administration of elemental sulfur]. AB - The guinea-pigs protractedly exposed per os to sulphur in a daily dose of 200 mg/kg body weight show, depending on the exposure period, in their livers, varied intensity of histochemical enzymatic reactions and different ALAT activity in blood serum. After 30 days' exposure to sulphur the intensity of enzymatic reactions is almost similar to that in controls or somewhat lower, whereas in the blood serum there is an increase in the activity of alanine aminotransferase. After the lapse of 90, 180 and 270 days of exposure, the animals' livers are observed to disclose a rise in reactions to the activity of mitochondrial, lysosomal enzymes as well as G-6-P-ase, esterase AS and ATP-ase, whereas in blood serum AspAT and ALAT reveal no difference in the activity level with regard to the control value. PMID- 6298565 TI - Cholinergic, adrenergic, and PGE1 effects on cyclic nucleotides and growth in cultured corneal epithelium. AB - Effects of adrenergic and cholinergic drugs and prostaglandin E1 on cyclic nucleotide accumulation and parameters of growth and basement membrane synthesis were examined in corneal epithelial cell cultures. 8-bromo-cGMP significantly (p less than 0.05) enhanced incorporation of labeled thymidine and leucine, as did acetylcholine and carbamylcholine, which elevated cGMP and decreased cAMP/cGMP ratio. Responses to acetylcholine were abolished by atropine and alpha bungarotoxin. Precursor incorporation was inhibited by dibutyryl cAMP and adenosine 5'-monophosphate and by norepinephrine, epinephrine, prostaglandin E1, and theophylline, which significantly elevated cAMP levels and cAMP/cGMP ratio. Propranolol, but not phenoxybenzamine, blocked responses to effective concentrations of norepinephrine. Norepinephrine, PGE1, and dibutyryl cAMP also significantly elevated uptake of labeled glucosamine and incorporation of labeled proline into collagenase-sensitive protein or the hydroxyproline fraction of protein hydrolysates, while acetylcholine had no effect on parameters of basement membrane synthesis. Propranolol blocked responses to norepinephrine. Results were consistent with a cGMP-mediated stimulatory role of the cholinergic transmitter in corneal epithelial growth regulation, cAMP-mediated beta-adrenergic suppression of regrowth and increased basement membrane production after initial injury to the corneal epithelium, and potentiation of the adrenergic effect by prostaglandins. PMID- 6298566 TI - Evidence for thyroid hormone receptors in uterine nuclei. AB - Nuclear preparations from the rat uterus contain specific, high affinity binding sites for T3, with the properties expected of a thyroid hormone receptor. The Kd value for the binding of T3 to these sites is 9.3 +/- 0.1 x 10(-10) M and the maximum number of binding sites is 0.11 +/- 0.02 pmoles T3 bound per mg of DNA. Competition experiments with Triac, D-T3, T4, and rT3 compared to L-T3 revealed relative affinities of 87%, 22%, 15%, and 1.6% respectively. T3 binding sites solubilized from uterine nuclear preparations by high salt extraction sediment at 3.6 S and are destroyed largely by trypsin treatment. Estradiol treatment of ovariectomized rats for 3 days did not alter the quantity of T3 binding sites expressed on the basis of DNA content. These results suggest that uterine nuclei contain a specific T3 receptor and raise the possibility that this hormone may have direct effects on the uterus. PMID- 6298567 TI - Importance of increased urinary calcium excretion in the development of secondary hyperparathyroidism of patients under glucocorticoid therapy. AB - Parathyroid function and calcium metabolism were studied in 44 patients under glucocorticoid therapy (steroid group) and in 25 control subjects. Nephrogenous cAMP and serum immunoreactive parathyroid hormone levels in the steroid group were significantly higher than those in control subjects (p less than 0.001). Nephrogenous cAMP in the steroid group correlated positively with prednisolone dosage (r = 0.424, p less than 0.01), and most patients who showed obvious elevations of nephrogenous cAMP had received over 10 mg/day of prednisolone for at least 2 mo. Fasting urinary calcium in the steroid group [166.1 +/- 78.5 (+/- SD) mg/g creatinine] was about 2 times greater than that in control subjects (74.1 +/- 35.6) (p less than 0.001). Fasting urinary calcium in control subjects correlated negatively with nephrogenous cAMP (r = -0.486, p less than 0.02). In contrast, these values in steroid group showed significant positive correlation (r = 0.631, p less than 0.001), suggesting that increased urinary calcium excretion is an important factor in the development of secondary hyperparathyroidism. Elevated nephrogenous cAMP and serum immunoreactive parathyroid hormone levels decreased after the administration of trichlormethiazide and/or 1 alpha hydroxy-vitamin D3. We conclude that increased urinary calcium excretion plays an important role in the development of secondary hyperparathyroidism in patients under glucocorticoid therapy and that the administration of thiazide and/or vitamin D could improve the secondary hyperparathyroidism caused by glucocorticoid therapy. PMID- 6298568 TI - Discrimination of multiple forms of phosphoprotein phosphatase in bovine thyroid. AB - Phosphoprotein phosphatases (phosphoprotein phosphohydrolase, EC 3.1.3.16) were partially purified from bovine thyroid with phosphorylated mixed histones, H1 histone and casein as substrates. Utilizing DEAE-cellulose chromatography, (NH4)2SO4 precipitation, gel filtration before and after freeze-thawing in 0.2 M 2-mercaptoethanol and histone-Sepharose chromatography, four fractions of enzyme activity were obtained and were designated as phosphatases I, IIA, IIB, and III. Phosphatases I had an apparent molecular weight of 155,000 and was dependent on Mn2+ for maximal activity. The enzyme had the greatest activity with histone H1 and was greatly stimulated by NaCl with phosphohistones as substrate. Phosphatases IIA and IIB had a molecular weight of about 70,000, were stimulated over 5-fold by Mn2+ and had much higher activities with phosphohistones than with casein in the presence of the cation. Phosphatase III, a possible catalytic subunit of larger molecular weight forms, had an apparent molecular weight of 30,000, was generally independent of Mn2+ and had high activities using all three substrates. Phosphatases I, IIA, and III were inhibited in a dose-dependent manner by sodium pyrophosphate (PPi), ATP, potassium phosphate (Pi) and sodium fluoride (NaF) when they were added directly to the reaction mixture with phosphorylated mixed histones as substrate. PPi was the most potent inhibitor and phosphatase III was the most sensitive to inhibition. PPi, ATP and NaF probably inactivated phosphatase III activity by removing an essential metal ion. After extensive dialysis to remove these inhibitors, the inactivated enzyme could be fully activated by Mn2+, but not by Mg2+, Ba2+, Cu2+, Cd2+, Ca2+, Zn2+ and Fe2+. Whereas the enzyme pretreated with Pi retained about 80% activity after dialysis, its activity was not further stimulated by Mn2+. The inactivated (demetallized) enzyme was less reactivated by Mn2+ in the presence of mM concentration of Pi. Moreover, the Mn2+-reactivated enzyme was again inactivated by Pi, NaF and ATP. Among them Pi was the most potent inactivator. These results suggest that Pi may have another inhibitory effect on metal ion binding besides on substrate binding and also that phosphatase III might be a metalloenzyme. In bovine thyroid, there are at least two major phosphoprotein phosphatases which may have different properties. Metal ion stimulation of phosphatase I and IIA activities may be through an interaction with the substrate or with a metal ion binding site on the regulatory subunit. The lowest molecular weight enzyme (phosphatase III) probably does not exist naturally in the cell. PMID- 6298569 TI - Sequence homology and recombination between the genomes of morphologically dissimilar bacteriophages LP 52 and theta. AB - A restriction fragment map of Bacillus licheniformis temperate phage LP 52 DNA (molecular weight 38.5 X 10(6)) was established, using restriction endonucleases BamHI (8 target sites), BglI (10 sites,) BglII (13 sites) and EcoRI (22 sites). The map is linear, with well-defined ends, without any signs of circular permutation. The DNA of a related phage, LP 51, produced identical restriction fragments. At least 62% DNA of LP 52 has been found homologous to the DNA of the recently discovered, morphologically quite dissimilar, phage I, as demonstrated by hybridization of electrophoretically separated restriction fragments of DNA. Under the same conditions, the DNAs of LP 52 and of the morphologically similar Bacillus subtilis phage phi 105 did not cross-hybridize. The homologous regions in the genomes of phages LP 52 and I have been shown to be colinear. Comparison of the cleavage maps of phages LP 52 and I has shown that, within the regions of homology, not a single restriction fragment and few restriction sites have been conserved during divergent evolution. Three major regions of heterology were defined; the longest one, covering the right-hand end of the map (73 +/- 2.75% up to 100% LP 52 genome length) appeared to contain genes coding for structural proteins of the virions; a shorter region at the left-hand end of the map (coordinates zero to 10.3 +/- 3.3% LP 52 genome length) and a very short central region (coordinates 41.8-43.9%) could be identified, the latter apparently containing a regulatory locus responsible for the heteroimmune behavior of the two phages. Recombinants between phages LP 52 and I were isolated. Mapping of recombinant genomes has indicated mutual substitution of allelic pieces of LP 52 and I DNAs upon strict conservation of overall genome length. PMID- 6298570 TI - Reduced [3H]-tetrodotoxin binding in the napts paralytic mutant of Drosophila. AB - The temperature-sensitive paralytic mutant of Drosophila, napts, has been shown to have defects in axonal physiology which suggest that it codes for a component of the voltage-sensitive sodium channel. Ligand binding studies using tritiated tetrodotoxin, a sodium channel antagonist, show a decrease in the apparent number of sodium channels in napts, providing additional support for this hypothesis. PMID- 6298571 TI - Genome organization in Xiphophorus (Poeciliidae; Telostei). PMID- 6298572 TI - Units of genetic expression in the virulence region of a plant tumor-inducing plasmid of Agrobacterium tumefaciens. AB - The effect of a large number of Tn3 insertions in the vir region of the Ti plasmid pTiA6NC on the virulence of Agrobacterium was determined. The Vir- insertions were mapped in three of the five loci that have been defined previously. Merodiploid Rec- strains carrying one insertion mutation on the Ti plasmid and another insertion mutation (or the homologous wild-type region) on a compatible plasmid were constructed and used in complementation tests for virulence in test plants. This analysis has revealed that there are ten units of gene expression, presumably transcription units in the vir region. Mutation in one of these units is confirmed to be dominant while those in all others are recessive. Co-infection of test plants with pairs of insertion mutants did not restore virulence. PMID- 6298573 TI - Chromosomal location and cloning of the gene (trmD) responsible for the synthesis of tRNA (m1G) methyltransferase in Escherichia coli K-12. AB - The trmD gene, which governs the formation of 1-methyl-guanosine(m1G) in transfer ribonucleic acid (tRNA), has been located by phage P1 transduction at 56 min on the chromosomal map of Escherichia coli. Cotransduction to tyrA at 56 min is 80%. From the Clarke and Carbon collection a ColE1-tyrA+ hybrid plasmid was isolated, which carried the trmD+ gene and was shown to over-produce the tRNA(m1G)methyltransferase. By subcloning restriction enzyme fragments in vitro, the trmD+ gene was located to a 3.4 kb DNA fragment 6.5 kb clockwise from the tyrA+ gene. The mutation trmD1, which renders the tRNA(m1G)methyltransferase temperature-sensitive both in vivo and in vitro could be complemented by trmD+ plasmids. These results suggest that the gene trmD+ is the structural gene for the tRNA(m1G)methyltransferase (EC 2.1.1.3.1). PMID- 6298574 TI - The structural gene (trmD) for the tRNA(m1G)methyltransferase is part of a four polypeptide operon in Escherichia coli K-12. AB - The trmD gene, which is the structural gene for the tRNA(m1G)-methyltransferase, is shown to be part of a polycistronic operon. A 4.6 kb SalI-EcoRI chromosomal DNA fragment contains the trmD gene (Bystrom and Bjork 1982). Subclonings, deletion mapping and Tn5 insertions into plasmid pBY03 have established the gene organization of the trmD area on the Escherichia coli chromosome. The different plasmid derivatives were analysed for expression of protein products using the minicell system. Such analyses established the organisation of genes encoding six polypeptides to be SalI1-48 K-13 K-25 K-31 K-15 K-16 K-EcoRI1. The 31 K polypeptide was shown to be the tRNA(m1G)methyltransferase. The trmD operon encodes for four polypeptides; 13 K-25 K-31 K(trmD)-15 K and the direction of transcription is from 13 K (promoter proximal) to 15 K (promoter distal). However, there might be a weak internal promoter between the trmD gene and the gene encoding the 15 K product. The level of expression from this operon in the minicell system does not seem to follow normal polarity since we observed high expression of 13 K, 25 K, and 15 K products but low expression of the internal trmD gene. PMID- 6298575 TI - DNA sequence change of a deletion mutation abolishing attenuation control of the threonine operon of E. coli K12. PMID- 6298576 TI - The cya locus of Escherichia coli K12: organization and gene products. AB - The cya gene region of Escherichia coli K12 has been cloned into plasmid pBR322. Detailed analysis of the locus, using in vitro recombination techniques as well as specific labelling of gene products has given information on the organization and products of the region. The cya gene is preceded by two DNA segments which behave as promoters of equivalent strength; a divergent, very strong, promoter is also present in the same control region. The former pair of promoters direct transcription of the cya gene which is expressed as a 95,000 dalton polypeptide; the latter promoter controls synthesis of a 40,000 dalton polypeptide. The cya gene product can complement a cya delta mutation when fully expressed or when expressed as a protein truncated at its carboxy terminal end. PMID- 6298578 TI - Recombination between short direct repeats in a recA host. AB - Spontaneous deletion derivatives of plasmid pHV14, a recombinant of pBR322, have been isolated in the recA strain HB101. About 2.5 kilobases (Kb) of DNA has been lost in each of the four plasmids described including, in two cases the region involved in control of copy number and the initiation of replication. Sequence analysis of each deletion junction has revealed that in 3 out of 4 cases the deletion event occurred by recombination between short direct repeats of as little as 7 base pairs (bp). PMID- 6298577 TI - Characterisation of the promoters for the ompA gene which encodes a major outer membrane protein of Escherichia coli. AB - The regulatory region of the ompA gene from Escherichia coli has been characterized by biochemical and genetic approaches. Two overlapping promoters, P1 and P2, organized in that order with respect to the ompA coding sequence, were identified and it was found that ompA possesses an unusually long leader region. Both P1 and P2 were active in an in vitro transcription system although S1 mapping analysis of the ompA mRNA made in vivo showed that P2 was mainly responsible for transcription of the gene. Confirmation of this was obtained by studying down-promoter mutants of ompA cloned in pSC101. These mutants were classified into two groups, deletions and insertions. The deletions, which were caused by the IS102 insertion element found in pSC101 removed the--35 regions of both P1 and P2. However, since P2 was distally situated with respect to the IS element it was less extensively damaged and it is proposed that the residual P2 sequence is responsible for the low level of expression observed. In addition to an IS102 insertion in the promoter region four IS1 insertion mutants were characterized. These had integrated at different positions in the ompA leader region and were all incompletely polar. PMID- 6298579 TI - Promoter mapping and DNA sequencing of the F plasmid transfer genes traM and traJ. AB - The nucleotide sequence of the DNA encoding the traM, finP and the promoter proximal segment of the traJ gene of the F plasmid has been determined. The predicted amino acid sequence for the traM protein shows that this inner membrane protein contains no signal sequence. The promoters for both the traM and traJ genes have been mapped by in vitro transcription and nuclease S1 protection experiments. No unambiguous location can be assigned to the finP gene but all candidates, if translated, would encode small proteins of between 24 and 52 amino acids. PMID- 6298580 TI - Characterization of a new, continuous rat fibroblast cell line, RFA-1: a high producer of interferon. AB - A new continuous rat fibroblast cell line (RFA-1) was characterized and its interferon (IFN) production and sensitivity to IFN compared with eight rat cell lines. After 95 passages, and about 380 population doublings, these cells exhibited chromosomal heterogeneity, but 55% still possessed the diploid chromosome number of 42. RFA-1 grown in Dulbecco's modified Eagle's medium-10% calf serum have a doubling time of 17.2 h and a saturation density of 141,487 cells/cm2. The geometric mean IFN production by these cells was 10(3.8) U/ml, and this was at least 10(0.5) U/ml higher than IFN production by the second highest IFN producer. Similarly, RFA-1 cells were at least three times more sensitive to the protective effects of interferon against vesicular stomatitis virus-induced cytopathic effects. PMID- 6298582 TI - Effect of dermonecrotic toxin of Bordetella pertussis on the spleen of CFW and C57BL/10ScN mice. PMID- 6298581 TI - Comparison of biological activities of Bordetella pertussis mutants lacking the ability to produce filamentous hemagglutinin with similar activities of other smooth strains. PMID- 6298583 TI - Superprecipitation-like phenomenon and destruction induced by adenosine 5' triphosphate in spirosomes isolated from Lactobacillus brevis. PMID- 6298584 TI - Severe chronic mucocutaneous candidiasis. Favourable response to oral therapy with ketoconazole. AB - Severe, early-onset, chronic mucocutaneous candidiasis was associated with bronchiectasis, oesophageal stricture, short stature and delayed puberty in a male aged 18 years. Topical treatment with antifungal agents, and several courses of intravenously administered transfer factor, amphotericin, and miconazole achieved only minor or transient improvement in the patient's condition. Correction of iron deficiency anaemia did not lead to alleviation of candidiasis. Skin reactivity to Candida antigen was absent and T-lymphocytes, which responded normally to phytohaemagglutinin (PHA), poke-weed mitogen and concanavalin A, had negative macrophage-inhibiting factor (MIF) and blastogenic responses to Candida antigen. Treatment with the orally effective imidazole derivative, ketoconazole, produced improvement within three days and clearing of mucosal lesions within five weeks. The patient then entered puberty spontaneously at the age of 20 years. After 18 months of treatment with ketoconazole, without side effects, the clinical manifestations of mucocutaneous candidiasis have not recurred. Because of the possibility of continuous, long-term administration, ketoconazole represents the best currently available agent for the treatment of chronic mucocutaneous candidiasis. PMID- 6298585 TI - Marijuana and apathy. PMID- 6298586 TI - The physics of proton NMR. AB - The physics of pulse NMR which is pertinent to an understanding of proton NMR imaging has been condensed and directed toward the medical physicist. The basic physical principals of spin manipulations using rf pulses are presented, and the relation between the quantum mechanical and the classical descriptions is covered in a rigorous fashion. The physics of relaxation is described and the relaxation times T1 and T2 are explained in some detail. Application of these spin manipulation techniques is illustrated by showing how they may be used in creating an image. PMID- 6298587 TI - Modification of the 50% maximum dose depth for 41-MeV (p+,Be) neutrons by use of filtration and/or transmission targets. AB - Several target configurations for the 41-MeV (p+,Be) reaction have been evaluated for the characteristics of the radiation field produced; depth dose, dose rate per microA, From analysis, it is concluded that to achieve the desired 13.2-cm depth for 50% of maximum dose and acceptable dose rate at a target-to-skin distance (TSD) of 125-150 cm, the neutron spectra must be filtered to preferentially absorb the lower-energy neutrons. Further increases in depth of 50% of maximum dose and a significant reduction in beryllium heating problems result if a partial transmission target is used with the terminal 30% of proton energy being deposited in a copper target backing. PMID- 6298588 TI - Characteristics of A-150 plastic-equivalent gas in A-150 plastic ionization chambers for p(66)Be(49) neutrons. AB - The evaluation of a gas mixture having an atomic composition similar to that of A 150 tissue-equivalent (TE) plastic has been extended to a high-energy neutron therapy beam. "A-150" gas, air, and methane-based TE gas were each flowed through A-150 plastic-walled ion chambers of different sizes and irradiated with p(66)Be(49) neutrons. A tentative value for W(A-150) of 27.3 +/- 0.5 JC-1 was derived for this beam. The W value of the A-150 gas mixture is compared to those of methane-based TE gas and of air for the p(66)Be(49) neutron beam as well as to corresponding values found in similar experiments using 14.8-MeV monoenergetic neutrons. PMID- 6298590 TI - [Rare case of a fibrous histiocytoma located in the parotid]. PMID- 6298591 TI - [Carotid chemodectoma]. PMID- 6298592 TI - Implementing the 1990 prevention objectives: summary of CDC's seminar. PMID- 6298594 TI - Soluble protein kinase fractions from DEAE-cellulose chromatography. A comparison between brain and heart from the rat. AB - The cytosol fraction from rat midbrain was chromatographed on DEAE-cellulose with a linear NaCl gradient (0-0.3 M). Two peaks of protein kinase activity were obtained when assayed with either histone or casein. A similar elution profile of the kinase activity was obtained from rat heart. The first peaks from midbrain and heart were compared in terms of their dependency upon cAMP and sensitivity to the endogenous protein kinase inhibitor. Neither of the two substances had an effect on the activity of the brain kinase. Furthermore, the dissociability of the midbrain and heart enzymes in the presence of cAMP or histone was compared by DEAE-cellulose chromatography. The heart enzyme was dissociated into a catalytic subunit characteristic of a cAMP-dependent protein kinase, whereas the brain kinase was totally unaffected by the cAMP or histone. The results of these tests indicate that although the elution profiles from DEAE-cellulose are similar between midbrain and heart, the first peak from brain contains a protein kinase that appears to be cAMP independent. PMID- 6298593 TI - Hydrogen peroxide mediated killing of bacteria. AB - Polymorphonuclear leukocytes (PMN) or neutrophils have multiple systems available for killing ingested bacteria. Nearly each of these incorporates H2O2 indicating the essential nature of this reactive oxygen intermediate for microbicidal activity. Following ingestion of bacteria by PMN, H2O2 is formed by the respiratory burst which consumes O2 and generates H2O2 from O2 .-. H2O2 is deposited intracellularly near bacteria within phagocytic vacuoles where it can react with the MPO-H2O2-halide system to form toxic hyperchlorous acid (HOCl) and/or possibly singlet oxygen (1O2). H2O2 can also react with O2 .- and/or iron (Fe++) from lactoferrin or bacteria to form the highly toxic hydroxyl radical (.OH). These mechanisms appear important since deficiencies of H2O2 production, myeloperoxidase or lactoferrin frequently increases their owner's susceptibility to infection. In particular, examination of PMN from infection prone patients with chronic granulomatous disease (CGD) most clearly demonstrates the importance of H2O2 in killing of bacteria. CGD PMN lack the capacity to effectively generate H2O2 and subsequently have impaired ability to kill catalase positive (H2O2 producing) but not catalase negative (not H2O2 producing) bacteria. PMN also have catalase and glutathione peroxidase systems in their cytoplasms to protect themselves from the toxicity of H2O2. Finally, while H2O2 is critical for host defense, it can also be released extracellularly and thereby play a significant role in PMN mediated tissue injury. PMID- 6298595 TI - Isolation and characterization of DNA-binding peptides from the serum: inhibition of transcription and comparison with the tissue peptides. PMID- 6298589 TI - Papovaviral persistent infections. PMID- 6298596 TI - Sequence comparison in the crossover region of an oncogenic avian retrovirus recombinant and its nononcogenic parent: genetic regions that control growth rate and oncogenic potential. AB - NTRE 7 is an avian retrovirus recombinant of the endogenous nononcogenic Rous associated virus-0 (RAV-0) and the oncogenic, exogenous, transformation-defective (td) Prague strain of Rous sarcoma virus B (td-PrRSV-B). Oligonucleotide mapping had shown that the recombinant virus is indistinguishable from its RAV-0 parent except for the 3'-end sequences, which were derived from td-PrRSV-B. However, the virus exhibits properties which are typical of an exogenous virus: it grows to high titers in tissue culture, and it is oncogenic in vivo. To accurately define the genetic region responsible for these properties, we determined the nucleotide sequences of the recombinant and its RAV-0 parent by using molecular clones of their DNA. These were compared with sequences already available for PrRSV-C, a virus closely related to the exogenous parent td-PrRSV-B. The results suggested that the crossover event which generated NTRE 7 took place in a region -501 to 401 nucleotides from the 3' end of the td-PrRSV parental genome and that sequences to the right of the recombination region were responsible for its growth properties and oncogenic potential. These sequences included a 148-base pair exogenous-virus-specific region that was absent from the RAV-0 genome and the U3 region of the long terminal repeat. Since the exogenous-virus-specific sequences are expected to be missing from transformation-defective mutants of the Schmidt-Ruppin strain of RSV, which, like other exogenous viruses, grow to high titers in tissue culture and are oncogenic in vivo, we concluded that the growth properties and oncogenic potential of the exogenous viruses are determined by sequences in the U3 region of the long terminal repeat. However, we propose that the exogenous-virus-specific region may play a role in determining the oncogenic spectrum of a given oncogenic virus. PMID- 6298597 TI - DNA sequence analysis of a mouse pro alpha 1 (I) procollagen gene: evidence for a mouse B1 element within the gene. AB - In a 3.8-kilobase mouse DNA sequence encoding amino acid sequences for the pro alpha 1(I) chain of type I procollagen, 14 coding sequences were identified which specify a sequence 95% homologous to amino acid residues 568 to 963 of the bovine alpha 1(I) chain. All of these coding sequences were flanked by appropriate splice junctions following the GT/AG rule. These observations suggest, but do not prove, that this pro alpha 1(I) gene is transcriptionally active. Of the 14 coding sequences, 7 were 54 base pairs in length, whereas the remainder were higher multiples of 54 base pairs. Nonrandom utilization of codons pertained throughout all of the coding sequences showing a preference (56%) for U in the wobble position. Two of the intervening sequences encoded imperfect vestiges of coding sequences which exhibited a codon preference different from that of the pro alpha 1(I) gene proper and were not flanked by splice junctions. One intervening sequence encoded a member of the mouse B1 family of middle repetitive sequences. It was flanked by 8-base-pair direct repeats and had a truncated A rich region, suggesting that it may be a mobile element. Within this element were sequences which could function as a RNA polymerase III split promoter. PMID- 6298598 TI - Patterns of integration of DNA microinjected into cultured mammalian cells: evidence for homologous recombination between injected plasmid DNA molecules. AB - We examined the fate of DNA microinjected into nuclei of cultured mammalian cells. The sequence composition and the physical form of the vector carrying the selectable gene affected the efficiency of DNA-mediated transformation. Introduction of sequences near the simian virus 40 origin of DNA replication or in the long terminal repeat of avian sarcoma provirus into a recombinant plasmid containing the herpes simplex virus thymidine kinase gene. (pBR322/HSV-tk) enhanced the frequency of transformation of LMtk- and RAT-2tk- cells to the TK+ phenotype 20- to 40-fold. In cells receiving injections of only a few plasmid DNA molecules, the transformation frequency was 40-fold higher after injection of linear molecules than after injection of supercoiled molecules. By controlling the number of gene copies injected into a recipient cell, we could obtain transformants containing a single copy or as many as 50 to 100 copies of the selectable gene. Multiple copies of the transforming gene were not scattered throughout the host genome but were integrated as a concatemer at one or a very few sites in the host chromosome. Independent transformants contained the donated genes in different chromosomes. The orientation of the gene copies within the concatemer was not random; rather, the copies were organized as tandem head-to tail arrays. By analyzing transformants obtained by coinjecting two vectors which were identical except that in one a portion of the vector was inverted, we were able to conclude that the head-to-tail concatemers were generated predominantly by homologous recombination. Surprisingly, these head-to-tail concatemers were found in transformants obtained by injecting either supercoiled or linear plasmid DNA. Even though we demonstrated that cultured mammalian cells contain the enzymes for ligating two DNA molecules very efficiently irrespective of the sequences or topology at their ends, we found that even linear plasmid DNA was recruited into the concatemer by homologous recombination. PMID- 6298599 TI - Transcriptional control of gene expression during development of Dictyostelium discoideum. AB - During development of the cellular slime mold Dictyostelium discoideum, approximately 2,000 to 3,000 regulated mRNAs are induced when amoebae enter multicellular aggregates. We used in vitro transcription in isolated nuclei to follow the synthesis of individual mRNA precursors during development; these were quantitated by hybridization to cloned cDNAs or genomic DNAs. Those RNAs that are present at all stages of development--the common RNAs--were transcribed by nuclei from cells at all stages of development. By contrast, those RNAs that are present only after cells begin to aggregate--here called aggregation stage RNAs--were transcribed only by nuclei from cells at the aggregation and postaggregation stages of development. The temporal pattern of in vitro transcription correlated well with the time course of accumulation of different aggregation stage mRNAs. Continued expression of aggregation stage genes normally depends upon cell-to cell contact or cyclic AMP (cAMP); when cells are disaggregated, the regulated mRNAs are rapidly and specifically degraded. When cAMP is subsequently added to the disaggregated cells, most of the mRNAs reaccumulate. We show here that disaggregation reduced 2- to 10-fold the relative transcription of several aggregation stage RNAs, whereas addition of cAMP to disaggregated cells reinduced the level of regulated gene transcription to values approximating those found in normal postaggregation cells. These results indicate that a representative set of Dictyostelium aggregation stage genes are under transcriptional control; both the transcription and the stability of these mRNAs require either continued cell-to cell interactions or cAMP. PMID- 6298600 TI - Alterations of gene structure in ethyl methane sulfonate-induced mutants of mammalian cells. AB - To determine the types of alterations in gene structure induced by DNA-alkylating agents, we analyzed the restriction enzyme cleavage patterns of adenine phosphoribosyltransferase gene sequences in mutant strains of Chinese hamster ovary cells deficient in this enzyme. Base pair changes as detected by loss of restriction enzyme sites were found, but no major internal gene rearrangements could be detected. PMID- 6298601 TI - Simian virus 40 and polyoma virus induce synthesis of heat shock proteins in permissive cells. AB - During the lytic infection of monkey and mouse cells with simian virus 40 and polyoma virus, respectively, the preferentially increased synthesis of two host proteins of 92,000 and 72,000 Mr was observed by 15 to 20 h after infection besides the general stimulation of most cellular proteins. The incubation of uninfected monkey and mouse cell cultures for 30 to 60 min at 43.5 degrees C induced the enhanced synthesis of at least three proteins of 92,000, 72,000 and 70,000 Mr, the last one being the major heat shock protein of mammalian cells. Two-dimensional gel electrophoresis and partial proteolytic digestion confirmed that the same 92,000- and 72,000-Mr proteins are stimulated by virus infection and thermal treatment. In simian virus 40-infected CV-1 cells, we also observed the weak stimulation of a 70,000-Mr protein comigrating in gel electrophoresis with the major heat shock protein. The 92,000-, 72,000- and 70,000-Mr proteins of monkey cells are structurally very similar to the corresponding proteins of mouse cells. In immunoprecipitations, no specific association of these proteins to simian virus 40 T antigens was noticed. PMID- 6298602 TI - Altered cell spreading in cytochalasin B: a possible role for intermediate filaments. AB - Trypsinized chicken embryo dermal fibroblasts plated in the presence of cytochalasin B (CB) quickly attached to the substrate and within 24 h obtained an arborized morphology. This morphology is the result of the pushing out of pseudopodial processes along the substrate from the round central cell body. There were no microfilament bundles in the processes of these cells plated in the presence of CB; however, the processes were packed with highly oriented, parallel aligned intermediate filaments. Only a few scattered microtubules were seen in these processes. These results demonstrated that in CB, cells are capable of a form of movement, i.e., the extension of pseudopodial processes, without the presence of the microfilament structures usually associated with extensions of the cytoplasm and pseudopodial movements. We also found that arborization did not depend on fibronectin since cells plated in CB did not have fibronectin fibers associated with the processes. Chicken fibroblasts transformed with tsLA24A, a Rous sarcoma virus which is temperature sensitive for pp60src, formed arborized cells with properties similar to those of uninfected fibroblasts when plated in the presence of CB at the nonpermissive temperature (41 degrees C). At the permissive temperature for transformation (36 degrees C), the cells attached to the substrate but remained round. These round cells were not only deficient in microfilament bundles but also lacked the highly organized intermediate filaments found in the processes of the arborized cells at 41 degrees C. Although both microfilament bundles and the fibronectin matrix were decreased after transformation with Rous sarcoma virus, neither was involved in the formation of processes in normal cells plated in CB. Therefore, the inability of the transformed cells to form or maintain processes in CB must be the result of another structural alteration in the transformed cells, such as that of the intermediate filaments. PMID- 6298603 TI - In vivo catenation and decatenation of DNA. AB - We have noted previously that when circular, but not linear, DNA or chromatin was injected into Xenopus laevis oocytes, much of it went through an intermediate form in which it did not readily enter an agarose gel; after a few hours, it reappeared as monomer DNA that had acquired its full complement of nucleosomes (T. J. Miller and J. E. Mertz, Mol. Cell. Biol. 2:1581-1593, 1982). We determined, using electron microscopy and a variety of biochemical techniques, the structure of this aggregated material. Most of it was oligomeric and multimeric catenanes of the injected sample. In addition, injection of DNA that had been catenated in vitro with DNA gyrase resulted in the conversion of most of it back to monomer circles. These findings demonstrate directly that both catenation and decatenation of DNA occur in vivo under physiological conditions. Whether these reactions play a crucial role in nucleosome formation, as well as in DNA replication and recombination, remains to be determined. PMID- 6298604 TI - Expression of hepatitis B virus surface antigen gene in cultured cells by using recombinant plasmid vectors. AB - By using a new host-vector system, expression of the gene coding for hepatitis B surface antigen has been studied. A subgenomic fragment of cloned hepatitis B viral DNA was inserted into the plasmid vector pSV010. Transfection of COS cells with the recombinant plasmid vector containing hepatitis sequences leads to the synthesis of hepatitis B surface antigen, which is released in the culture medium in the form of 22-nm particles similar to those found in the sera of hepatitis carriers. PMID- 6298605 TI - Characterization of transposable element-associated mutations that alter yeast alcohol dehydrogenase II expression. AB - Seven cis-dominant, constitutively expressed mutations of the normally glucose repressible isozyme of alcohol dehydrogenase (ADHII) from the yeast Saccharomyces cerevisiae are caused by insertion of transposable elements from the Ty1 family in front of the ADHII structural gene (ADR2) (V. M. Williamson, E. T. Young, and M. Ciriacy, Cell 23:605-614, 1981). We cloned ADR2 with its associated Ty1 element from five S. cerevisiae strains carrying these mutations. Comparison of the Ty1 elements by heteroduplex studies and restriction enzyme analyses indicated that four were very similar; the fifth, although the same size as the others (about 5.6 kilobases), differed by the presence of two large substitutions of approximately 1 and 2 kilobases. The DNA sequences of the terminal direct repeats (deltas) were very homologous but not identical and were similar to previously reported Ty1 element direct repeats. We determined the 5'-flanking sequences of the ADR2 gene isolated from a wild-type strain and from five Ty1 associated mutations. The 5-base pair target sequence at the site of Ty1 insertion was present at both ends of each Ty1 element. The sites of insertion of the elements were all different and occurred from 125 to 210 base pairs in front of the coding region of ADR2. The 5' end of the major transcript as determined by S1 mapping was the same in wild-type cells and in Ty1-associated constitutive mutants and was approximately 54 base pairs upstream from the coding region. ADR2 transcripts were not detected when a solo delta sequence was present in the 5' flanking region of this gene. PMID- 6298606 TI - Construction and use of a dominant, selectable marker: a Harvey sarcoma virus dihydrofolate reductase chimera. AB - The transcriptional promoter of the Harvey sarcoma virus long terminal repeat has been used to construct a biologically active dihydrofolate reductase chimera. The construction placed the long terminal repeat at the 5' end of a dihydrofolate reductase cDNA. This chimera mediated methotrexate resistance when introduced into wild-type NIH3T3 mouse cells by transfection. The chimeric sequences were expressed in the form of polyadenylated RNA and dihydrofolate reductase protein and were amplified when the methotrexate-resistant transfectants were selected to grow in increasing methotrexate concentrations. This chimera was dominant acting and able to confer a methotrexate-resistant phenotype on wild-type NIH3T3 cells. It has been used in cotransfection experiments with DNA from human tumor cells to obtain foci of methotrexate-resistant transformed NIH3T3 cells resulting from uptake of exogenous DNA. The transfected methotrexate-resistant cells carried double minute chromosomes that appeared to contain DNA acquired during transfection. PMID- 6298607 TI - Plasmid-directed synthesis of hepatitis B surface antigen in monkey cells. AB - We introduced the gene encoding the hepatitis B virus surface antigen (HBsAg) into simian virus 40 (SV40)-based plasmids capable of autonomously replicating in both Escherichia coli and permissive monkey cells. After introduction into monkey cells by transfection, these plasmids directed the synthesis of high levels of HBsAg, as determined by immunofluorescence, radioimmunoassays, and identification by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the polypeptides comprising the antigen. Expression was dependent upon the presence of an SV40 promoter, with both the early and late promoters able to effectively initiate transcription. Using expression of HBsAg to assay promoter function, we demonstrated that an intact copy of the SV40 72-base pair repeat, which constitutes an essential element of the SV40 early promoter during the lytic SV40 cycle and which can enhance the transcriptional activity of heterologous promoters, was not required for HBsAg expression, suggesting that the hepatitis genome contains an enhancer element capable of complementing that provided by the 72-base pair repeat element of SV40. The antigen appears to be glycosylated after synthesis in transfected cells and is apparently secreted, as evidenced by the localization of [35S]cysteine-labeled antigen to the medium of transfected cultures. Using constructions in which the first ATG sequence appearing in HBsAg mRNA was that corresponding to the gene encoding the mature form of the antigen, we demonstrated that these post-translational events could occur without the involvement of a putative precursor peptide suggested by the DNA sequence of the viral genome. In view of the inability of hepatitis B virus to propagate in vitro, this strategy offers a convenient approach for further characterizing the biosynthesis of this antigen and may provide a means to identify additional polypeptides encoded by this virus. PMID- 6298608 TI - Some lymphoid cell lines transformed by Abelson murine leukemia virus lack a major 36,000-dalton tyrosine protein kinase substrate. AB - Fibroblasts transformed by Abelson murine leukemia virus differ from normal fibroblasts in that they contain several cellular proteins, including one of 29 and one of 36 kilodaltons, which are phosphorylated at tyrosine residues. Since it has been shown before that these proteins also become phosphorylated at tyrosine after transformation of fibroblasts by a number of other retroviruses, their phosphorylation may play an important role in the transformation of these cells. In contrast, the 36-kilodalton phosphoprotein was not detectable in three of the four lines of Abelson virus-transformed B lymphoma cell lines studied here. These three cell lines, RAW307.1.1, 18-48, and 18-81, and a B lymphoma induced by mineral oil, WEHI 279, were all found to lack both the phosphorylated and unphosphorylated forms of the 36-kilodalton protein. It thus appears that expression of this major cell protein is not essential for the survival of B lymphoma cells in culture and that the phosphorylation of the 36-kilodalton protein at tyrosine is not essential for transformation of pre-B lymphocytes by Abelson virus. PMID- 6298609 TI - Interaction between the Rous sarcoma virus transforming protein and two cellular phosphoproteins: analysis of the turnover and distribution of this complex. AB - The transforming protein of Rous sarcoma virus (RSV), pp60src, was previously shown to associate with two cellular proteins of Mr 90,000 and 50,000 in RSV transformed chicken cells. In this report, we demonstrate that this interaction is specific for a discrete population of pp60src molecules. Newly synthesized pp60src was found to preferentially associate with pp90 and pp50 to form a short lived complex. The half-life of this complex varied from 9 to 15 min in cells transformed by nondefective strains of RSV. This interaction between pp60src, pp50, and pp90 took place in a soluble fraction of the cell, and the complex bound pp60src molecules were not phosphorylated on tyrosine. These results suggest that pp90 and pp50 may be involved in the processing of pp60src molecules before the association of pp60src with the plasma membrane. The kinetics of dissociation of this complex were shown to be altered in cells infected with viruses containing a temperature-sensitive defect in the src gene. When cells infected with these viruses were grown at the nonpermissive temperature, more than 90% of the pp60src molecules were associated with pp90 and pp50, and little or no dissociation was observed in a 3-h chase period. These results suggest that mutations in the src gene which affect the transforming activity of pp60src also affect the stability of the interaction of pp60src with pp90 and pp50. PMID- 6298610 TI - Allergenic cross-reactivity of cytochromes c of Kentucky bluegrass and perennial ryegrass pollens. AB - The allergenic cross-reactivity of cytochromes c isolated from Kentucky bluegrass (KBG) and ryegrass (RG) pollens was demonstrated by the finding that both cytochromes elicited PCA reactions of comparable titers in rats sensitized with murine reaginic antiserum to either KBG or RG cytochrome c. However, allergenic differences were revealed at limiting doses of the cytochromes c; under these conditions RG cytochrome c elicited PCA reactions only with the homologous reaginic serum, whereas KBG cytochrome c elicited PCA reactions with both murine reaginic sera. Moreover, in experiments involving inhibition of PCA, RG cytochrome c failed to neutralize some of the IgE antibodies of the antiserum to KBG cytochrome c, whereas KBG cytochrome c neutralized all IgE antibodies of both murine reaginic antisera. From these results it may be concluded that: (1) the two cytochromes c share common allergenic determinants, and (2) the KBG cytochrome c possesses additional allergenic determinant(s) not present on RG cytochrome c. The radioallergosorbent test, utilizing KBG cytochrome c discs and a pool of sera of individuals allergic to KBG, was inhibited to the extent of 87 or 41% by the addition of 10 micrograms of KBG or RG cytochrome c, respectively. By contrast both cytochromes c at 10 micrograms were equally effective in the inhibition (92%) of the binding of the IgE antibodies in this serum pool to RG cytochrome c allergosorbent discs. These experiments confirmed that the two cytochromes share common allergenic determinants. PMID- 6298611 TI - [Effect of antacids on mineral metabolism in persons with healthy kidneys. Double blind study using an antacid containing magnesium aluminum silicate hydrate]. PMID- 6298612 TI - [Hypoglycemia]. PMID- 6298613 TI - Mutagenic repair in Escherichia coli. VIII. Effect of gyrB mutations on ultraviolet light mutagenesis. AB - Introduction into Escherichia coli WP2 bacteria of a mutation in the gyrB locus previously shown to reduce the degree of chromosomal superhelicity caused a small decrease in the frequency of UV-induced mutations to streptomycin resistance (but not significantly) and to tryptophan independence (mostly ochre suppressors) in excision repair-proficient bacteria. It did not influence the 'broth effect' or the rate or extent of 'mutation frequency decline' of suppressor mutations. In an excision-deficient (uvrA 155) background the yield of UV-induced streptomycin resistant mutations was lower in gyrB bacteria at all doses; the yield of tryptophan-independent mutations was slightly lower at low doses and slightly higher at high doses. In both excision-proficient and -deficient bacteria the yield of UV-induced mutations to rifampicin resistance was apparently lower in gyrB mutants but this could be due at least in part to a hypersensitivity of some Rifr gyrB bacteria to UV. The number of spontaneous tryptophan-independent mutations was lower in gyrB bacteria but this was almost certainly due to their poorer viability on tryptophan-limiting plates and not to a lower spontaneous mutation rate. In a temperature-sensitive presumed gyrase-deficient strain a small decrease in mutant yield at low doses was observed following incubation at restrictive temperature before UV. This was ascribed to an enhancement of excision repair. Our failure to find any significant effect of gyrB mutations does not support the hypothesis that hairpin formation (which should be dependent on a high degree of superhelicity) is involved in determining the 'broth effect', 'mutation frequency decline' or the probability that a mutation will occur spontaneously. Dramatic effects of superhelicity on UV mutagenesis also seem to be unlikely. PMID- 6298614 TI - Establishment by SV40 transformation and characteristics of a cell line of xeroderma pigmentosum belonging to complementation group F. PMID- 6298615 TI - Short-term tests for transplacentally active carcinogens. A comparison of sister chromatid exchange and the micronucleus test in mouse foetal liver erythroblasts. AB - The effectiveness of 6 chemicals (benzo[a]pyrene, (BaP), cyclophosphamide (CP), diethylnitrosamine (DEN), methyl methanesulphonate (MMS), mitomycin C (MC) and procarbazine (PC) ) as inducers of micronuclei in foetal liver and maternal bone marrow erythroblasts has been determined, and related to that of gamma-radiation. CP, DEN, MMS and PC were all more effective in the foetal liver. The induction of micronuclei and SCEs by each chemical in foetal erythroblasts after in vivo exposure was measured. When expressed as induction of sister-chromatid exchanges (SCEs) per erythroblast/induction of micronuclei per erythroblast (/microM/kg), the ratios obtained were MC 580, BaP 470, DEN 430, CP 258, MMS 140 and PC 13. The lowest doses detected as potentially genotoxic by each test in foetal liver erythroblasts are (with the exception of PC which is a relatively ineffective inducer of SCEs) similar. When isolated foetal livers were exposed in vitro, SCE dose responses to BaP, MC, MMS and PC could be directly related to those from in vivo exposure, indicating the role of the foetal liver in metabolic activation, but CP was considerably more cytotoxic. The transplacental micronucleus test, and in vivo/in vitro method for SCEs in foetal liver erythroblasts, provide sensitive, complementary assays for genotoxic effects of chemicals during prenatal life. Since foetal liver possesses greater metabolic potential than adult bone marrow, the transplacental tests respond to genotoxic agents not detected by bone-marrow systems. PMID- 6298616 TI - Toxicological implications of the organic fraction of aerosols: a chemist's view. PMID- 6298617 TI - Genotoxicity of aerosol extracts. Some methodological aspects and the contribution of urban and industrial locations. AB - The genotoxicity of aerosol extracts was investigated with the Ames test and an SCE test in vitro. In addition to experiments to evaluate current techniques, a so-called gradient study was carried out in an industrialized part of The Netherlands to obtain insight into the contribution of urban and industrial locations to the genotoxicity of aerosol extracts. The influence of variations in the sampling was investigated. The results indicate that moderate variations in the volume of air sampled per unit of time and of the duration of the sampling period do not have a great influence on the effect measured. Experiments with filters impregnated with 14C-labelled benzo[a]pyrene showed that this compound is converted during sampling into directly active mutagens; no evaporation occurred. The results of experiments to evaluate the extraction procedure indicated that an 8-h Soxhlet extraction with methanol is better than, or as good as, Soxhlet extraction with other solvents. The gradient study showed that the aerosol downwind from industrial/urban areas exerted a reproducibly stronger effect in the Ames test and the SCE test when compared with aerosol from sea air. PMID- 6298619 TI - Petriellidiosis of the skin. PMID- 6298618 TI - Enhanced survival of ultraviolet-irradiated herpes simplex virus in cells exposed to antiviral agents. AB - Enhanced survival of UV-irradiated HSV-1 is demonstrated in monkey cells exposed to inhibitors of viral DNA synthesis. Phosphonoacetic acid (PAA), adenine arabinoside (ara-A), and cytosine arabinoside (ara-C) pretreatment of infected cells is associated with concentration-dependent reactivation of UV-HSV-1. At concentrations that result in enhanced virus survival, inhibition of cell DNA synthesis is observed by either ara-A or ara-C, but not by PAA. Pretreatment of uninfected cells with acycloguanosine (ACG) is not associated with reactivation of irradiated HSV-1, and this is probably due to insufficient generation of ACG triphosphate, the active inhibitor of viral and cell DNA synthesis. PMID- 6298620 TI - The effect of ketoconazole on the phagocytosis and intracellular killing of Candida albicans by polymorphonuclear granulocytes. PMID- 6298621 TI - Phosphatidate phosphatase activity in the blood fluke, Schistosoma mansoni. PMID- 6298622 TI - An adult case of type Ib glycogen-storage disease. Enzymatic and histochemical studies. PMID- 6298623 TI - Health policy report. The Centers for Disease Control. PMID- 6298624 TI - Naloxone administration releases catecholamines. PMID- 6298625 TI - A tale of two islets. PMID- 6298626 TI - Facilitated induction of hippocampal long-lasting potentiation during blockade of inhibition. PMID- 6298627 TI - Activation of nif gene expression in Azotobacter by the nifA gene product of Klebsiella pneumoniae. AB - Structural similarity of nitrogenase, the enzyme responsible for biological nitrogen fixation, from various diazotrophic bacteria has been shown by intergeneric biochemical complementation of component proteins in vitro, DNA and protein sequence analysis, and DNA hybridization between nif (nitrogen fixation) structural genes from Klebsiella pneumoniae and genomic sequences from other nitrogen-fixing bacteria. Despite nitrogenase homologies, little is known about similarities among nif regulatory mechanisms although repression of nitrogenase synthesis by NH4+ and O2 occurs in most diazotrophs. In K. pneumoniae, the ntr (gln) genes concerned with regulation of nitrogen metabolism control expression of the nifLA operon whose products act as repressor (nifL) and activator (nifA) of the seven other nif transcriptional units. Here we report that the nifA gene product of K. pneumoniae can activate expression of nif genes in both Azotobacter vinelandii and Azotobacter chroococcum, organisms whose aerobic physiology contrasts with that of the facultative K. pneumoniae. PMID- 6298628 TI - Structure of a genomic clone encoding biologically active human relaxin. AB - Relaxin is a peptide hormone synthesized in the corpora lutea of ovaries during pregnancy and is released into the blood stream prior to parturition. Its major biological effect is to remodel the mammalian reproductive tract to facilitate the birth process. Determination of the structure of human relaxin is thus a first step in opening up the possibility of clinical intervention in cases of difficult labour. However, the limited availability of human ovaries during pregnancy has prevented both direct amino acid sequence determination and isolation of cDNA clones obtained from relaxin producing tissue. Our approach has therefore been to screen directly for a human relaxin gene using an homologous porcine relaxin cDNA probe. We report here the successful identification of a genomic clone from which the structure of the entire coding region of a human preprorelaxin gene has been determined. Synthesis of biologically active relaxin has shown that the novel gene structure described herein codes for an authentic human relaxin. We believe this is the first successful synthesis of a biologically active hormone whose structure was predicted solely from the structure of a genomic clone. PMID- 6298630 TI - Cyclic AMP and development in the slime mould. PMID- 6298629 TI - Mitochondrial DNA sequences in the nuclear genome of a locust. AB - The endosymbiotic theory of the origin of mitochondria is widely accepted, and implies that loss of genes from the mitochondria to the nucleus of eukaryotic cells has occurred over evolutionary time. However, evidence at the DNA sequence level for gene transfer between these organelles has so far been limited to a single example, the demonstration that a mitochondrial ATPase subunit gene of Neurospora crassa has an homologous partner in the nuclear genome. From a gene library of the insect, Locusta migratoria, we have now isolated two clones, representing separate fragments of nuclear DNA, which contain sequences homologous to the mitochondrial genes for ribosomal RNA, as well as regions of homology with highly repeated nuclear sequences. The results suggest the transfer of sequences between mitochondrial and nuclear genomes, followed by evolutionary divergence. PMID- 6298631 TI - Possible role of acetylcholinesterase in regulation of postsynaptic receptor efficacy at a central inhibitory synapse of Aplysia. AB - Most of the effects of acetylcholinesterase (AChE) on synaptic transmission are considered to be related to its acetylcholine (ACh) hydrolysing properties. This is clearly apparent from changes which occur in the characteristics of the miniature endplate potential and of the endplate potential at neuromuscular junctions when AChE is inhibited1-4 and during the development of enzymatic AChE activity at maturing synapses5. However, we report here that after inhibiting AChE in a cholinergic synapse in Aplysia, we found an increase not only in postsynaptic responses to presynaptic stimulation and to ionophoretic application of ACh on postsynaptic receptors, but also to ionophoretic application of carbachol. This could not be explained by the inhibition of the ACh hydrolysing function of the enzyme, as carbachol is not hydrolysed by AChE. A possible explanation of these observations is that inhibition of the enzyme affects a property of the ACh receptor (AChR) itself. PMID- 6298632 TI - Identification and nucleotide sequence of a human locus homologous to the v-myc oncogene of avian myelocytomatosis virus MC29. AB - Avian myelocytomatosis virus MC29 is a replication-defective acute leukaemia virus which induces a variety of tumours in chickens including sarcomas, renal and hepatic carcinomas, and myelocytomatosis. The oncogenic potential of the virus is mediated by the gene v-myc, acquired from sequences (c-myc) present in normal uninfected chicken DNA. Sequences closely related to chicken c-myc have been highly conserved throughout evolution, from Drosophila to vertebrates. The hypothesis that c-myc may be involved in neoplastic transformation has been strengthened by the finding that B-cell lymphomas induced in chickens by avian leukosis virus (ALV) are often associated with increased expression of c-myc resulting from integration of the ALV provirus adjacent to the c-myc gene. More recently, it has been demonstrated that the malignant human cell line HL-60, derived from the peripheral blood leukocytes of a patient with acute promyelocytic leukaemia, expresses elevated levels of myc-related mRNA associated with an amplification of the c-myc gene. To explore the relationship of the human cellular myc gene with the corresponding viral oncogene from MC29, and to provide a framework for the analysis of the mechanism and significance of c-myc amplification in human tumours, we have isolated and determined the nucleotide sequence of a genomic clone prepared from a normal human library which contains all domains sharing homology with v-myc. PMID- 6298633 TI - Corrections to the nucleotide sequence of the src gene of Rous sarcoma virus. PMID- 6298634 TI - The oncogenic potential of herpes simplex viruses: evidence for a 'hit-and-run' mechanism. AB - Experiments to determine the mechanism of transformation of herpes simplex virus (HSV) have identified fragments of viral DNA which are able to initiate transformation. No set of viral genes seems to be consistently retained or expressed in the transformed cells or in human cervical tumours, suggesting that viral DNA is not needed to maintain the transformed phenotype. In fact there is no conclusive evidence that initiation of neoplasia is mediated by a viral protein. Here we revisit the 'hit-and-run' hypothesis and its implications for HSV-induced tumorigenicity. PMID- 6298635 TI - Complete nucleotide sequences of the T24 human bladder carcinoma oncogene and its normal homologue. AB - DNA sequence analysis of the activated oncogene from the T24 human bladder carcinoma line and two alleles of its normal cellular progenitor (c-Ha-ras-1) indicates that the genes encompass at least four exons, and that only a single point mutation residing within the first exon distinguishes the coding region of both alleles of the normal gene from their activated counterpart. Both versions of the gene encode a protein which is predicted to differ from the corresponding viral gene product at three amino acid residues, one of which was previously shown to represent the major site of phosphorylation of the viral polypeptide. PMID- 6298636 TI - An immunologically active chimaeric protein containing herpes simplex virus type 1 glycoprotein D. AB - Herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2) cause both persistent and latent infections, including recurrent cutaneous disease, lethal neonatal disease, central nervous system disease and other clinical syndromes. Modified live vaccines or conventionally prepared subunit vaccines have generally been unsuccessful in the treatment of HSV-1 and HSV-2 infections from the standpoints of safety and efficacy. It has been established that HSV-1 and HSV-2 infectivity may be neutralized in vitro with antisera directed specifically against each of the four major glycoproteins of the virus (gA/gB, gC, gD and gE) and antisera against glycoprotein gD, of either HSV-1 or HSV-2, are capable of neutralizing both HSV-1 and HSV-2 infectivity in vitro and in vivo. We have previously reported on the identification, DNA sequence and expression at low level in Escherichia coli of the gD gene of HSV-1 strain Patton. Here we describe construction of a hybrid gene encoding a chimaeric protein containing HSV-1 gD, bacteriophage lambda Cro and E. coli beta-galactosidase (gD-beta-gal) protein, which is expressed at high level in E. coli. Moreover, the chimaeric protein elicits antibodies in rabbits that not only immunoprecipitate gD from cells infected with HSV-1 and HSV-2 but also neutralize HSV-1 and HSV-2 infectivity in vitro. PMID- 6298637 TI - Is ribonucleotide reductase the transforming function of herpes simplex virus 2? AB - Transformation of cells by herpes simplex virus 2 (HSV-2) can be induced by the BglII C (0.43-0.58 map units) or N (0.58-0.625) fragments of the viral genome. Sequences partially overlapping both fragments (0.566-0.602) encode two 3' coterminal mRNAs; these in turn direct the synthesis of two related polypeptides of molecular weight 140,000 (140K) and 35K (refs 4, 7), which may be involved in transformation. Recently, a temperature-sensitive (ts) mutation affecting HSV induced ribonucleotide reductase has been mapped within this common region (B.M. Dutia, personal communication). We have partially purified the induced reductase and raised a rabbit antiserum to it which inhibits the enzyme activity and immunoprecipitates from infected cells a 144K polypeptide and minor species including a 38K polypeptide. Here we show that a monoclonal antibody to the putative transforming proteins competes with the rabbit serum for the 144K and 38K antigens and also immunoprecipitates specifically the induced reductase activity. These results suggest a possible role for ribonucleotide reductase in HSV-2-induced transformation. PMID- 6298638 TI - Characterization of a human colon/lung carcinoma oncogene. AB - DNA sequences capable of inducing oncogenic transformation of NIH3T3 mouse cells are found in a number of human tumour cell lines. When DNAs of these cell lines are applied to monolayer cultures of the mouse fibroblasts, foci of transformed cells are observed 2-3 weeks later. DNA from cells of such primary foci can be used in turn to induce foci in a second cycle of gene transfer. The human DNA sequences responsible for transformation have been called oncogenes, the best characterized of which is closely related to the Harvey murine sarcoma virus oncogene. Here we present a characterization of an oncogene which we found originally to be present in DNA of the SW480 colon carcinoma cell line. We indicate its structural outlines and demonstrate, in extension of reported results, its presence in an activated form in the genome of several types of human tumour cell lines as well as in biopsy tissue from an adenocarcinoma of the large bowel. We identify this tumour oncogene with c-Ki-ras2, one of two known members of the Kirsten ras family of human proto-oncogenes, extending a series of recent reports which have demonstrated homologies between human oncogenes and those of Harvey and Kirsten murine sarcoma viruses. The c-Ki-ras2 oncogene of several tumour cell lines is shown to be amplified. PMID- 6298639 TI - Retroviruses and transposable elements--which came first? PMID- 6298640 TI - Only membrane-associated RSV src proteins have amino-terminally bound lipid. PMID- 6298641 TI - Histopathological definition of the adult and calf types of bovine leukosis. AB - Histopathological distinctions were examined on 36 cattle 2 months to 12 years of age affected with leukosis. The distribution of tumors pointed out previously as the characteristic of the adult type of leukosis was observed in 27 adult cases, which, except two, were positive for the immunodiffusion test of bovine leukemia virus (BLV). The noticeable development of massive and lumpy tumors in serosa, loose connective tissue, and lymph nodes seemed to suggest a neoplastic accumulation in areas related to the lymphatic way. Splenic involvement and intrasinusoidal leukotic immigration in the liver were present in cases with the leukemic manifestation of peripheral blood. The neoplastic involvement of bone marrow, however, was difficult to find. On the other hand, in 5 calves and 4 adolescent or adult cattle, the neoplastic proliferation was observed throughout such hematopoietic organs as lymph nodes, bone marrow, liver, spleen, thymus and tonsil. The hepatic lesion was characterized by the neoplastic proliferation in the portal triads No BLV antibody could be detected from these cattle, except one adult. As the conclusion of the present study, the tumor of adult type regarded as the pathologic manifestation of enzootic bovine leukosis is defined as malignant lymphoma (non-Hodgkin's lymphoma or lymphosarcoma) of undifferentiated , histiocytic-, or poorly- differentiated lymphocytic- cell type. And lymphocytic leukemia seems to be desirable term to distinguish the lymphoid malignancy regarded as the characteristic of the calf type from that of the adult type. PMID- 6298642 TI - Experimental studies on vertical infection of mice with Japanese encephalitis virus III. Effect of gestation days at the time of inoculation on placental and fetal infection. AB - An experiment was carried out on the effect of gestation days at the time of inoculation on the establishment of experimental vertical infection with Japanese encephalitis virus in mice. In it, mice of the CFW strain in a closed colony were inoculated intravenously with a field strain at different times over a period from 3 to 12 days of gestation. After that, an attempt was made to recover the virus from the placenta and fetus to estimate the rate of infection. As a result, placental infection was established not when the virus was inoculated at 3 days of gestation, but when it was inoculated at 4 days of gestation or later. The rate of infection was relatively high when the virus was inoculated at 6 days of gestation or later. Fetal infection was established relatively frequently when the virus was inoculated some time between 7 and 10 days of gestation, but quite infrequently when the virus was inoculated at any other time than this. There was a difference in rate between placental and fetal infection at a given time of inoculation in days of gestation. This difference seemed to have been induced not by a difference in intensity of viremia appearing after inoculation, but by a difference in degree of development between placental and fetal tissues at the time of inoculation. PMID- 6298643 TI - [The Nobel Prize for Medicine 1982. On essential fatty acids, prostaglandins and acetylsalicylic acid]. PMID- 6298644 TI - Two cellular proteins that are precipitated by an antitumor-RSV rat serum. AB - Here we report studies on non-viral proteins specific for transformation precipitated by immune sera. These sera were from rats injected by PR-RSV transformed rat XC cells and from rabbits immunized by subcutaneous injections of cytoplasmic membranes from rat cells transformed by Fujinami avian sarcoma virus. Immune complexes were analyzed in one- or two-dimensional gel electrophoresis and by autoradiography. Only by using antisera obtained from rats bearing XC-cells induced tumors, two proteins were identified with an apparent molecular weight (Mr) of 105 000 (p105) and 98 000 (p98) and an isoelectric point of 6.4 and 6.2, respectively, in extracts of mammalian cells transformed by avian sarcoma viruses (ASV). These proteins were common to all ASV-transformed cell lines. In kinase reaction, the protein p98 can be phosphorylated. Similar proteins were neither detected in normal nor in chemically transformed cells. None of the proteins were precipitated with control serum from rat, rabbit, and goat. PMID- 6298645 TI - Quantitative changes of tropomyosin synthesis in RSV-transformed mammalian cells in comparison with normal fibroblasts. AB - Protein profiles of three tumor mammalian cell lines and three control cell lines were analyzed by two-dimensional gel electrophoresis. Comparison of the cells labeled metabolically with 35S-methionine has revealed obvious differences in an area of molecular weights 34 000--36 000 daltons and pH 4.5-5.0. Two spots have been proposed to be alpha- and beta-tropomyosin subunits. Their synthesis decreases after cell transformation. PMID- 6298646 TI - Auditory brainstem responses in the aged cat. AB - Auditory brainstem responses (ABRs) were compared in young adult and aged cats. Mean thresholds for click-evoked ABRs were greater in the aged cats. Clicks normalized to 15 and 30 dB above individual thresholds at rates of 10, 20, 50 and 100/sec evoked ABRs with similar latencies and central conduction times in both groups. Background noise at equal intensity for all cats completely suppressed ABRs evoked by clicks 30 dB above threshold in 2/3 of the young but none of the old cats. As rise time of a 25 msec noise burst at equal intensity for all cats increased 1, 2, 5, and 10 msec, latency of wave 4 increased more for the old cats than for the young. Summed monaural ABRs from both ears were greater than binaural ABRs for waves 4 and 5 in both groups. These data indicate peripheral auditory dysfunction in aged cats but little abnormality in auditory brainstem transmission with click intensity normalized for ABR threshold. PMID- 6298647 TI - Aging in the rat medial nucleus of the trapezoid body. I. Light microscopy. AB - The medial nucleus of the trapezoid body (MTB), a cell group in the superior olivary complex, was examined in an age-graded series of rats for neuron loss, changes in the giant synaptic endings (chalices of Held) on MTB neurons, and accumulation of age pigment. Neuron counts were done on protargol-stained paraffin sections of MTBs from a series of 17 rats aged 2-3, 6, 18, and 24 months. Between 2-3 and 24 months a 34% decrease in the mean number of MTB neurons was observed. Significant loss (p less than 0.05) was first evident in the early portion of the life span, between 2-3 and 6 months. In thionin-stained sections, there was no change with aging in the proportions of three MTB neuron types: principal cells (approximately 82%), elongate cells (approximately 15%), and stellate cells (approximately 3%). In young adult rats, 25-26% of all MTB neurons were associated with identifiable chalices of Held in protargol stained sections. This ratio did not vary significantly with aging. Age pigment accumulation in the MTB was examined in 2 micrometers Araldite sections stained with toluidine blue. Age pigment deposits were larger and more numerous in the MTBs of old animals, but not as extensive as has been described previously in many other parts of the nervous system. This study is the first to report neuron loss in an animal brainstem nucleus. PMID- 6298648 TI - Age-related changes of benzodiazepine and GABA binding sites in the rat retina. AB - The changes in the number and sensitivity of benzodiazepine and GABA binding sites in the rat retina during postnatal development, adulthood and ageing and their functional relationship at different ages have been studied. Data indicate an increase in the total number of both GABA and benzodiazepine binding sites with age. In contrast, the activation of retinal benzodiazepine receptor binding by GABA is significantly reduced in aged rats with respect to young adult and newborn rats. Moreover, the activation of retinal benzodiazepine receptor binding induced by dark exposure of the animals is present in young adult rats but is lost in aged rats. These results suggest that in the retina of aged rats there is an increase of GABA and benzodiazepine receptors which have lost their functional connection. PMID- 6298649 TI - ACTH and corticosterone secretion following indomethacin, in intact, adrenalectomized and dexamethasone-pretreated male rats. AB - The present study was designed to determine the role of glucocorticoids in the mediation of the stimulatory effect of indomethacin (IM) on the hypothalamo hypophyseal-adrenal axis. Intact male rats were treated with a single injection of either dexamethasone (Dex; 20 micrograms/100 g body weight), IM (5 mg/100 g body weight), or IM + Dex or their respective vehicles. In Dex-treated rats, ACTH and corticosterone (CS) were significantly reduced over a period of 20 h as compared to the vehicle-treated group. As we have previously demonstrated, injection of IM markedly elevated serum ACTH and CS for at least 20 h. Administration of Dex 2 h prior to IM treatment delayed the stimulatory effect of IM for 5 h; subsequently, however, Dex was without effect. In adrenalectomized rats, ether stress elicited a marked rise in serum ACTH levels. On the contrary, IM was completely ineffective in these rats. These studies suggest that the mode of action of IM in causing hypersecretion of ACTH and CS is by interfering with the negative feedback effects of the glucocorticoids. PMID- 6298650 TI - Malignant optic glioma presenting radiologically as a "cystic" suprasellar mass: case report and review of the literature. AB - The diagnosis of malignant glioma of the optic nerves and chiasm may be difficult because these rare lesions may mimic other suprasellar lesions. We report a case of glioblastoma multiforme of the optic chiasm in which the appearance of the lesion on computed tomographic (CT) scan was consistent with that of craniopharyngioma or cystic pituitary adenoma. We suggest that malignant optic glioma should be considered in the differential diagnosis of an adult with progressive visual loss of rapid onset, even when ophthalmological examination strongly suggests extrinsic compression of the anterior optic pathways and when the CT scan apparently indicates a cystic mass lesion. PMID- 6298651 TI - High grade astrocytoma with spontaneous meningeal and cranial invasion. PMID- 6298652 TI - Familial brain tumors: studies of two families and review of recent literature. AB - We studied two families in each of which three or more individuals were affected by brain tumors. In the first family, which had no evidence of neurofibromatosis or tuberous sclerosis, a man, his sister, and her son developed histologically proven gliomas; the man's great uncle was historically reported to have died from a brain tumor, but the exact nature of the tumor was not known. In this family two of the tumors were low grade astrocytomas of the cerebrum, whereas the third was a mixed glioma of the cerebellum. Karyotypic analysis of this tumor showed no marker chromosomes. A second family had a history of an unusual concentration of brain tumors. In one patient the tumor was a histologically verified glioma. Four other patients had historically reported brain tumors, the descriptions of which suggested gliomas. Both families showed involvement of individuals in adjacent generations, although in both instances there were skipped generations. Twins, siblings, or parents and children are the kindred groups affected in most other reported families with multiple brain tumors. The mode of inheritance of brain tumors in these two families and recent literature on the conditions associated with familial brain tumors are discussed. PMID- 6298653 TI - Chronic rubidium does not prevent dopamine receptor supersensitivity. PMID- 6298655 TI - Effect of level of spatial synchronization of cortical potentials on spread of evoked responses to cortical electrical stimulation. PMID- 6298654 TI - Correlation between energy characteristics of the cortical theta rhythm and motor responses in rabbits. PMID- 6298656 TI - Morphological and functional types of visual cortical neurons in rabbits during postnatal development. PMID- 6298657 TI - Exploration of the facial nerve canal by high-resolution computed tomography: anatomy and pathology. AB - A correlative study of the anatomy and the radiological appearance of the intratemporal course of the facial nerve canal was carried out. Isolated temporal bones and temporal bones of cadaver heads were examined with thin-section high resolution computed tomography in the axial, coronal and Stenvers' projections, then sectioned with a microtome and the radiologic and anatomic images were correlated. Appropriate projections for visualization of each segment of the facial nerve canal on CT were established. The high-resolution CT Stenvers' projection proved very useful for visualization of the geniculate ganglion fossa, as also of the tympanic and mastoid segments in their full length. The high resolution CT appearance of lesions characteristic for each segment of the facial nerve canal are presented. PMID- 6298658 TI - The climbing fiber projection to the flocculus and adjacent paraflocculus in the cat. AB - The olivo-cerebellar projection to the flocculus and adjacent parts of the ventral paraflocculus was investigated by means of anterograde transport of tritiated leucine. Four subdivisions of the principal olive (caudal lateral bend, ventro-lateral outgrowth, rostral and caudal dorsal cap) and the rostral tip of the medial accessory olive were involved in the projection. In the flocculus contralateral to the injection site, eight different climbing fiber strips are distinguished, most of which extend into the most caudal lobule of the ventral paraflocculus and the area of cortex connecting it with the flocculus. One of these, present in the extreme caudal edge of the area under study, originated from the rostral part of the medial accessory olive and was labeled C2. The others originate in the principal olive and are labeled F-strips. Whereas the caudal lateral bend is connected with a single strip (F7), the remaining subdivisions give origin to two separated strips. The ventro-lateral outgrowth, the rostral and caudal dorsal cap are the respective sites of origin of the strips F2 + F5, F3 + F6 and F1 + F4. The significance of the strips is discussed in relation to the cerebellar relay nuclei, known to receive floccular efferents. A new nomenclature is proposed for the lobular subdivision of the flocculus and adjacent parts of the ventral paraflocculus, together with a simple method for the construction of a two-dimensional diagram of these areas. PMID- 6298659 TI - The neuronal architecture of the anteroventral cochlear nucleus of the cat in the region of the cochlear nerve root: horseradish peroxidase labelling of identified cell types. AB - Golgi impregnations of the posterior part of the cat's anteroventral cochlear nucleus have revealed two types of neurons, bushy cells with short bush-like dendrites and stellate cells with long, tapered processes; Nissl stains have revealed globular and multipolar cell bodies with dispersed and clumped ribosomal patterns, respectively. In the present study, we injected horseradish peroxidase into the trapezoid body. Ipsilaterally, retrograde, diffuse labelling of neurons, presumably through damaged fibers, yielded Golgi-like profiles of numerous bushy cells with typical dendrites and with thick axons projecting toward the trapezoid body. Stellate cells were almost never labelled in this way. Anterograde diffuse labelling of thick axons demonstrated calyx endings in the contralateral medial nucleus of the trapezoid body. In the electron-microscope, the perikarya of diffusely-filled bushy neurons were found to have the dispersed ribosomal pattern and the kinds of synaptic endings typical of globular cells, including large profiles of end-bulbs from cochlear nerve axons. After injections restricted to the medial trapezoid nucleus, granularly-labelled cells in the cochlear nucleus were almost completely confined to the contralateral side; Nissl counterstaining showed them to be globular cells in the posterior part of the anteroventral cochlear nucleus. After larger injections, involving surrounding regions of the superior olivary complex, granular labelling occurred throughout the ventral cochlear nucleus on both sides. There is also evidence that stellate cells in Golgi impregnations correspond to multipolar cell bodies in Nissl stains. We conclude that bushy cells typically correspond to globular cells, which receive end-bulbs from the cochlea and send thick axons to the contralateral medial trapezoid nucleus, where they form calyces on principal cells. Principal cells, in turn, are known to project to the lateral superior olive and to one of the nuclei of origin of the crossed olivo-cochlear bundle, which feeds back to the cochlea. In this circuit, correlations between synaptic patterns and particular physiological signal transfer characteristics can be suggested. These could be related to binaural intensity interactions in the lateral superior olive and to a regulatory loop involving the olivo-cochlear bundles. PMID- 6298660 TI - Ethylene oxide polyneuropathy. AB - Sensorimotor polyneuropathy developed in two workers who had been exposed to ethylene oxide gas repeatedly for several months. Sural nerve biopsies revealed axonal degeneration with mild changes of the myelin sheath. Unmyelinated fibers were also involved. Muscle biopsies showed typical denervation atrophy. Symptoms improved after exposure to ethylene oxide terminated. PMID- 6298661 TI - [Multiphase instrumental diagnosis of minimal breast cancer]. AB - The value of a breast surgery procedure based on clinical examination, mammography, telethermography, echotomography and cytological examination for the diagnosis of symptomatic or high risk patients was tested through a retrospective survey of 28 patients with minimal breast cancer. The data confirm that mammography is a much better clinical and instrumental test than other systems. When associated with radiography it gives definite, early diagnosis of more breast neoplasias than other methods, facilitating prognosis and reducing the number of biopsies necessary. PMID- 6298662 TI - [A sporadic case of vitamin D-resistant rickets in an adult]. AB - A case of probably sporadic vitamin D-resistant rickets is reported. The patient had suffered from rickets of average intensity in childhood, and the signs had persisted into his adult life. Serious symptoms due to phosphate depletion and marked osteomalacia, however, did not appear until he was 56. A brief description of the differential diagnosis and a summary of the literature on this uncommon form are followed by an account of the gradual biochemical and clinical improvement observed after treatment with phosphates and vitamin D. PMID- 6298663 TI - [Acute herpes simplex hepatitis with favorable outcome in an otherwise healthy adult]. AB - A non fatal case of Human Herpes Simplex hepatitis is presented, and the features of 15 previously reported cases are briefly reviewed. The patient, an otherwise normal middle-aged woman, presented as acutely ill, with typical mucocutaneous lesions, but recovered in a few days. Liver biopsy showed multiple focal areas of cellular necrosis with marked inflammatory infiltration, but without evidence of intranuclear eosinophilic inclusions. The possible underestimation of cases of Herpes Simplex hepatitis is proposed. PMID- 6298665 TI - Distribution and morphology of vasoactive intestinal polypeptide-like immunoreactive neurons in regio superior of the rat hippocampal formation. AB - The distribution and morphology of vasoactive intestinal polypeptide-like (VIP-L) immunoreactive neurons were studied in thick (70-120 microns) sections from the rat hippocampal formation using VIP antibody in combination with immunohistochemistry. Analysis of serial sections cut through the regio superior revealed a relatively dense aggregation of cells in stratum lacunosum-moleculare. Classification of cells in these (and other) layers on the basis of their morphology and orientation showed that hippocampal VIP-positive cells, unlike cortical ones, constitute a remarkably heterogeneous population. Studies of axonal trajectories of individual VIP cells showed that while some have local terminal arbors within the stratum lacunosum-moleculare close to the parent soma, other VIP cells send longer projections to other layers and subfields of the hippocampal formation. PMID- 6298664 TI - Long-lasting depression of parallel fiber-Purkinje cell transmission induced by conjunctive stimulation of parallel fibers and climbing fibers in the cerebellar cortex. AB - In rabbit cerebellar cortex, local stimulation of parallel fibers induced field potentials with two negative peaks, n1 representing conducting spikes of parallel fibers and n2 postsynaptic excitation in dendrites of Purkinje cells and other cortical cells. Conjunctive stimulation of parallel fibers and climbing fibers at 4 Hz for 30-120 sec caused a significant depression of n2 potential which lasted for at least 1 h. Such an effect could not be produced by stimulation of climbing fibers or parallel fibers alone. These observations support the plasticity assumption in the Marr-Albus model of the cerebellum. PMID- 6298666 TI - Presence of GABA receptors in rat oviduct. AB - A low concentration of high-affinity, saturable and specific [3H]GABA binding sites has been identified in a membrane fraction of rat oviduct. The specific binding of [3H]GABA was displaced by unlabelled GABA, muscimol and bicuculline. Furthermore in oviductal slices, GABA and a known GABA receptor agonist, i.e. muscimol, produced a significant elevation of cyclic AMP levels, which could be antagonized by GABA receptor blockers, i.e. picrotoxin and bicuculline. The present results indicate that GABA receptors may have a functional significance in rat oviduct. PMID- 6298667 TI - Cochlear potentials in the Bronx waltzer mutant mouse. AB - The Bronx waltzer mutant mouse has a unique cochlear abnormality in which the outer hair cells appear normal but the inner hair cells are absent. Potentials recorded from the round window indicate that the gross cochlear nerve action potential is very small or absent and cochlear microphonics are present but of small amplitude. Positive and negative summating potentials can both be recorded, indicating that mammalian outer hair cells are capable of producing both positive and negative DC potentials. PMID- 6298668 TI - Sub-miniature junction potentials in excitatory neuromuscular synapses of the locust. AB - By recording miniature excitatory junction potentials (mejps) intracellularly at two points from a multiterminally innervated muscle fibre it is possible to select mejps whose amplitudes are not substantially affected by electrotonic decay. Many amplitude histograms of such selected mejps from untreated locust jumping muscle show a bimodal distribution with a high proportion of small amplitude mejps (sub-mejps). Most amplitudes of excitatory junction potentials (ejps) resulting from the release of a single transmitter quantum correspond to the large-mode mejps. Tetanic nerve stimulation, in high [Mg2+]o without Ca2+, greatly reduces the proportion of sub-mejps. It is concluded that there are two modes of spontaneous transmitter release from the motor nerve endings. PMID- 6298669 TI - Evidence for the presence of specific receptors for insulin-like growth factors 1 (IGE-1) and 2 (IGF-2) and insulin throughout the adult human brain. AB - The present study examines adult human brain tissue for the presence of receptors for insulin-like growth factors (IGFs) 1 and 2 and insulin. Binding sites for all hormones were widely distributed throughout the brain removed at autopsy, with the highest specific binding being found in the various cortical regions. Cross reaction studies performed on frontal lobe biopsy material indicated specific binding sites for IGF-1 and insulin whereas iodinated IGF-2 was equally displaced by IGF-1 and IGF-2, suggesting the presence of an additional IGF-2-like binding site in the adult brain. PMID- 6298670 TI - Angiotensin-converting enzyme blockade by Captopril changes angiotensin II receptors and angiotensinogen concentrations in the brain of SHR-sp and WKY rats. AB - Angiotensin II-sensitive neurons in the brain of spontaneously hypertensive rats (SHR-sp) and of Wistar Kyoto rats (WKY) treated with the angiotensin-converting enzyme inhibitor Captopril were investigated for possible differences at receptor sites. Furthermore, the concentrations of angiotensinogen and renin were measured in different brain regions of these animals by biochemical assay. The higher receptor sensitivity of septal neurons to angiotensin II which existed in SHR-sp as compared to WKY was diminished by Captopril. Angiotensinogen concentrations were lower in the anterior hypothalamus but not in the septum of SHR-sp as compared to WKY. Captopril increased the level in both strains. Renin concentrations did not differ in SHR-sp and WKY. Chronic treatment with Captopril induced an increase of about 20% in septum and hypothalamic regions of SHR-sp and WKY rats. Whether these changes are causally linked to the hypertension in SHR-sp remains to be investigated. PMID- 6298671 TI - Purines interact with 'central' but not 'peripheral' benzodiazepine binding sites. AB - The effects of several purines and the purine uptake inhibitor, dipyridamole, on the binding, to rat brain membranes, of 4 benzodiazepines with different pharmacological specificities were studied. While all purines tested displaced the binding of [3H](+)-3-methyl-clonazepam and [3H]Ro15-1788, selective agonist and antagonist ligands respectively for 'central' benzodiazepine receptors, purines had little or no affinity for [3H]Ro5-4864 'peripheral'-type binding sites in brain, heart or kidney. These results suggest that purines interact with a pharmacologically relevant class of central benzodiazepine 'receptors', and not with central and peripheral 'acceptor' sites labelled by the benzodiazepine Ro5 4864. PMID- 6298672 TI - Olfactory connections of substantia innominata and nucleus of the horizontal limb of the diagonal band in the rat: an electrophysiological study. AB - Extracellular unit recordings performed on 113 spontaneously discharging neurons in the substantia innominata-nucleus of the horizontal limb of the diagonal band region revealed that 46 (41%) were transsynaptically discharged or inhibited following stimulation of the main and accessory olfactory bulb (MOB, AOB) and the prepyriform cortex (PPC). A significantly greater number of cells were responsive to MOB and PPC than to AOB shocks. Eight (7%) neurons were antidromically invaded following MOB volleys. These findings suggest that the MOB has functional reciprocal connections with the 'ventral striatum', a region which can function, presumably as a nodal point for convergence of visual, gustatory and olfactory information relevant to feeding and drinking behavior. PMID- 6298673 TI - Capsaicin-sensitive non-cholinergic excitatory innervation of the guinea-pig tracheobronchial smooth muscle. AB - Field stimulation of the isolated main bronchi of the guinea-pig results in a rapid contraction followed by a sustained contractile response. Tetrodotoxin abolished these effects. The first phase was strongly inhibited by hyoscine, indicating that it was mediated mainly by excitation of cholinergic nerves. The lasting contraction was abolished by capsaicin tachyphylaxis but it was resistant to the effects of hyoscine, hexamethonium or physostigmine. It is suggested that capsaicin-sensitive non-cholinergic nerves have major excitatory effect on the guinea-pig bronchial smooth muscle and there is also evidence for their influence on the trachea. PMID- 6298674 TI - Electrophysiological study with K+- and Ca2+-sensitive micropipettes of GABA receptors in the rat neurointermediate lobe in vitro. AB - An efflux of K+ and a decrease in extracellular Ca2+ activity are to be expected when GABA markedly depolarizes the membrane of unmyelinated axons or secretory cells. Accordingly, we used extracellular recordings of ionic movements to specify GABA receptor presence in parts of the pituitary having GABAergic innervation: the neurohypophysis and intermediate lobe (NIL). We identified a site of action having the same desensitization characteristics and pharmacological criteria as the GABA A receptor which modulates Cl- -conductance. Baclofen, a GABA B agonist, was without effect. The possible distribution of receptors and role of GABAergic synapses in modulating neurotransmitter and hormone release by the NIL are discussed. PMID- 6298675 TI - Circadian rhythms in electrical discharge of rat suprachiasmatic neurones recorded in vitro. AB - The discharge of suprachiasmatic (SCN), retrochiasmatic (RCA) and arcuate (ARC) neurones was recorded in brain slices incubated for periods up to 30 h. The firing rate of cells in the SCN, but not in the RCA and ARC, exhibited a circadian rhythm similar to that reported for the SCN in freely moving animals. This rhythm cannot be ascribed to subtle exogenous diurnal variations in the incubation conditions. It is concluded that the SCN explant is capable of endogenous generation of at least one circadian cycle in vitro. PMID- 6298676 TI - Response to pressure and vibration of slowly adapting cutaneous mechanoreceptors in the human foot. AB - Unit activity from slowly adapting cutaneous mechanoreceptors of the foot and of the tibial part of the leg was recorded in man using the microneurography method. All the receptors increased their discharge frequency when pressure was applied to the zone of maximum sensitivity of their receptive field. They were also very sensitive to mechanical vibrations of small amplitude (0.2-0.5 mm) applied precisely to their zone of maximum sensitivity. Some of them are able to respond one-to-one to vibration at frequencies up to 200 Hz. The conditions of activation of the slowly adapting receptors seem to exclude a major contribution to kinaesthetic effects induced by vibrations applied to the tendon of tibialis anterior and extensor digitorum longus muscles [12]. PMID- 6298677 TI - Response of adrenal efferent nerve activity to non-noxious mechanical stimulation of the skin in rats. AB - The effects of brushing various segmental skin areas (neck, lower chest, abdomen and thigh) on adrenal sympathetic efferent nerve activity were examined in anesthetized rats. In the CNS intact rats, brushing of all areas produced a large reflex decrease in the efferent nerve activity, whereas, in the spinalized rats, brushing the lower chest or abdominal skin area produced an increased response while brushing the neck or thigh skin area produced no significant changes in the efferent nerve activity. It is suggested that the segmental excitatory organization of the adrenal nerve reflex caused by brushing within the spinal cord is modified into a non-segmental or generalized inhibitory organization by the supraspinal structure. PMID- 6298678 TI - Alteration of high-affinity binding sites of neurotransmitter receptors in rats after neonatal exposure to streptomycin. AB - Central neurotransmitter functions were analyzed in dyskinetic rats that were treated subacutely with streptomycin as neonates. Data indicated that the treatment caused an increase in the binding of [3H]spiroperidol in striatum and of [3H]serotonin in frontal cortex, but no significant changes were observed in the binding of [3H]quinuclidinyl benzilate and [3H]muscimol to cerebellum and of [3H]diazepam to frontal cortex. The increase in the binding of [3H]spiroperidol and [3H]serotonin was evident in both sexes; however, the increase of [3H]spiroperidol binding was statistically significant only in females. Kinetic studies as revealed by Scatchard plots show that the increase in the binding of the two ligands is due to an increase in the number of the receptors. These results support the concept of a central mechanism of action for the streptomycin induced dyskinesias. PMID- 6298679 TI - Methylmercury-induced depression of neuromuscular transmission in the rat. AB - Effects of bath applied methylmercury on neuromuscular transmission were assessed in the isolated phrenic nerve-hemidiaphragm of adult male rats using conventional microelectrode recording techniques. At concentrations of 20 and 100 microM, methylmercury increased the frequency of miniature end-plate potentials (MEPPs) from control values of 0.3-0.8/sec to 1.5-35/sec. An increase in MEPP frequency occurred after 15-40 min of exposure to 20 microM, and 5-15 min after exposure to 100 microM methylmercury. At a concentration of 4 microM, methylmercury did not increase MEPP frequency, and in fact, slightly decreased it. The increase in MEPP frequency occurred more rapidly and was smaller in magnitude with normal Ca2+ and Mg2+ concentrations (2 mM and 1 mM, respectively) than with low Ca2+ and high Mg2+ (1 mM and 8 mM, respectively) in the bathing solution, although in each case, significant increases in MEPP frequency were observed. The increase in spontaneous neurotransmitter release caused by methylmercury was not reversed by treatment with 1 microM tetrodotoxin. MEPP amplitude was not significantly altered by any concentration of methylmercury tested. End-plate potentials (EPPs) evoked under conditions of diminished quantal content (1 mM Ca2+ and 8 mM Mg2+) were decreased in amplitude, and finally blocked by 20 microM and 100 microM methylmercury, but not by 4 microM methylmercury. EPP block occurred after 30-40 min with 20 microM methylmercury, and after 4-5 min with 100 microM methylmercury. At the time of EPP block, MEPPs of normal size were still observed. The amplitude of end-plate depolarizations produced by iontophoretic application of ACh remained constant during 20-30 min of exposure to 100 microM methylmercury. Quantal content of transmitter release and the immediately available store of neurotransmitter were drastically reduced by higher concentrations of methylmercury, while the probability of release was increased. Resting membrane potentials of muscle fibers were not altered significantly by any concentration of methylmercury tested. The observed effects of methylmercury were not reversible upon washing with drug free solution for one hour. It is suggested that acute methylmercury poisoning irreversibly alters presynaptic function at the mammalian neuromuscular junction. PMID- 6298680 TI - [Use of the complement fixation and passive hemagglutination tests in diagnosing chronic forms of ophthalmic adenovirus infection]. PMID- 6298681 TI - Effect of cytotoxic drugs on the function of the hypothalamo-pituitary-adrenal axis. AB - We measured the serum concentration of thyroid-stimulating hormone (TSH), its response to exogenic TSH-releasing hormone (TRH), as well as cortisol plasma levels before and after stimulation with adrenocorticotrophic hormone in 15 patients before, during and after a chemotherapeutic cycle. 3/6 patients receiving only cytotoxic drugs developed a marked suppression of the TSH response to TRH and 1 of these patients showed an impairment of the adrenal function under chemotherapy. This was also observed in 7/9 patients receiving both cytotoxic drugs and corticosteroids; however, the individual pattern of the impairment was quite variable. 5/9 patients in this group developed a suppression of the TSH response to TRH. Impairment of the hypothalamo-pituitary-adrenal axis function under cytotoxic drugs and/or corticosteroids occurs with great variability and the mechanisms involved in its etiology are not yet fully understood. PMID- 6298682 TI - Evaluation of coated and uncoated synthetic absorbable sutures in rabbit limbal wounds. AB - Polymer coatings have been applied to braided synthetic absorbable sutures to decrease tissue drag and improve knot tying characteristics. To examine the possibility that increased "slipperiness" would decrease knot security, coated (Dexon Plus) and uncoated (Dexon S) polyglycolic acid sutures were compared in rabbit limbal wounds. Both suture types had comparable knot security, tissue reaction, and absorption rates, while the coated sutures caused less tissue drag. Clinical trials of coated polyglycolic acid suture for limbal wounds are warranted. PMID- 6298683 TI - The acute retinal necrosis syndrome. Part 2: Histopathology and etiology. AB - The acute retinal necrosis syndrome is manifested by diffuse uveitis, vitritis, retinal vasculitis, and acute necrotizing retinitis (see Part 1). We studied the histopathology and electron microscopic findings of an eye enucleated from a 67 year-old man with typical acute retinal necrosis. Histology showed profound acute necrosis of the retina, retinal arteritis, and eosinophilic intranuclear inclusions in retinal cells. Electron microscopy demonstrated a herpes group virus in all layers of affected retina. The implications of these findings for antiviral and other treatments are discussed. PMID- 6298685 TI - [Herpesvirus antigens and antibodies in IgA glomerulonephritis]. PMID- 6298684 TI - Heteropolypeptides on Titan? AB - Since hydrogen cyanide is a component of Titan's hazy atmosphere, HCN polymers might also be present by way of a low energy pathway leading initially to the synthesis of polyaminomalonitrile. Subsequent reactions of HCN with the activated nitrile groups of this HCN homopolymer would then yield heteropolyamidines, readily converted to heteropolypeptides following contact with frozen water on the surface of Titan. Similar HCN polymers in the reducing atmospheres of Jupiter and Saturn could be major contributors to the yellow-brown-orange appearance of these giant planets. Any detection of such HCN chemistry by the Voyager missions or the pending Galileo probe would constitute evidence for the hypothesis that heteropolypeptides on the primitive Earth were synthesized directly from hydrogen cyanide and water without the intervening formation of alpha-amino acids. PMID- 6298686 TI - Glomus tumor surgery: the approach, results, and problems. PMID- 6298687 TI - The neurosurgeon in skull base operations. PMID- 6298688 TI - The results of the complement fixation test on the cytomegalovirus antibodies in children admitted to the Gadjah Mada University, Yogyakarta. PMID- 6298689 TI - Neuropathology of systemic malignant neoplasia. PMID- 6298690 TI - Wilms' tumor: review of ultrastructure and histogenesis. PMID- 6298691 TI - [Morphological evaluation of the material obtained by selective catheterization of the bronchial tree from patients with round lung shadows]. PMID- 6298692 TI - [Retrospective evaluation of 107 cases of cancer of the gallbladder by autopsy]. PMID- 6298693 TI - [Method of preparation of the material obtained by fine-needle aspiration biopsy for examination by scanning electron microscopy]. PMID- 6298695 TI - Pulmonary and systemic vascular actions of tolazoline in anesthetized dogs. AB - The cardiovascular actions of tolazoline are poorly understood. Therefore, we administered tolazoline (2 mg kg-1, IV) to anesthetized adult dogs and puppies. Tolazoline induced transient pulmonary and systemic pressor responses, transient systemic vasoconstriction and pulmonary vasodilation, and transient hypoxemia and acidosis in pentobarbital anesthetized dogs. None of these responses were evident during a second injection of tolazoline, indicating either the development of tachyphylaxis or a secondary antagonistic property of tolazoline. These responses to tolazoline were also prevented by alpha adrenergic blockade. Histamine H1 - and H2-receptor blockade sustained the transient pulmonary and systemic pressor and systemic vasoconstrictor responses to tolazoline. Thus, tolazoline appears to activate both alpha and histamine receptors in the pulmonary and systemic vascular beds of the anesthetized dog. In anesthetized puppies, tolazoline induced a slight pulmonary pressor response, marked bradycardia, and systemic hypertension. The present findings confirm the pharmacological complexity of tolazoline and suggest that anesthesia, age, and pulmonary vascular tone are important factors in determining the cardiovascular responses to tolazoline. PMID- 6298694 TI - Multiple benzodiazepine receptors in the ovine brain: ontogenesis, properties, and distribution of 3H-diazepam binding. AB - Benzodiazepine receptors in the ovine frontal cortex were present at 56 days gestation and developed slowly until 96 days when the number increased rapidly, reaching adult levels by 120 days gestation. Scatchard analysis of 3H-diazepam specifically bound to cortical membranes suggested high (KD approximately equal to 2.0 nM) and low (KD approximately equal to 20.0 nM) affinity benzodiazepine receptors at all stages of development. Whereas the affinity of these receptors for 3H-diazepam did not alter during development, the number of both high and low affinity receptors increased significantly between 56 and 120 days gestation. The number of low affinity receptors were higher in late gestation and early neonatal life than in adulthood. The functional state of these receptors as determined by sensitivity to GABA did not alter during development. However, in the adult, nitrazepam, flunitrazepam, midazolam, and 1-methylisoguanosine were more potent in displacing 3H-diazepam at the low affinity than the high affinity receptor, whereas chlordiazepoxide and diazepam had greater potency at the high affinity binding site. Development of the benzodiazepine receptor in the majority of other brain regions studied occurred primarily after 68 days gestation, as was the case in frontal cortex. In contrast, hindbrain and midbrain benzodiazepine receptors had reached adult levels by 68 days gestation. PMID- 6298696 TI - Radionuclide angiography: as diagnostic method for Wilms' tumor with direct extension into the heart. AB - Radioisotope angiography may be used as an initial study in Wilms' tumor. We report a patient who had a total nephrectomy but then had severe cardiovascular complications. These led to open heart surgery and removal of a tumor lodged in the right atrium. Radioisotope angiography, performed two weeks earlier, showed a "cold" image on the anterior wall of the right ventricle. This finding was later confirmed by other studies and finally at the time of a second operation. The purpose of this paper is to discuss the different methods employed in this case, to emphasize the risks attributed to each, and finally to suggest the use of non invasive studies such as radioisotope angiography as a practical method of study in infancy. PMID- 6298697 TI - Maternal diabetes does not alter postnatal development of the hepatic glucagon receptor-adenylate cyclase system in the rat. PMID- 6298698 TI - Live attenuated varicella vaccine: the KMcC strain in healthy children. AB - The KMcC strain of live, attenuated varicella-zoster virus vaccine was studied in healthy children as a preliminary step toward varicella vaccine studies with this strain in children with leukemia. Forty-three children were immunized: 26 with the 40th passage vaccine and 17 with the 50th passage. Studies included surveillance for clinical reactivity, oropharyngeal excretion of vaccine virus, viruria, and viremia. Antibody responses were assayed by fluorescent antibody to membrane antigens and immune adherence hemagglutination. Cell-mediated immune responses were assayed by lymphocyte proliferation to varicella-zoster virus specific antigens. There was 100% seroconversion to the KMcC passage 40 and 50 vaccines (by fluorescent antibody to membrane antigen assay). Every child studied developed in vitro lymphocyte proliferation to varicella-zoster virus antigens. Papular skin lesions, probably vaccine related, occurred in 31% of the 40th passage vaccinees but in only 6% of the 50th passage vaccinees. The 50th passage KMcC strain vaccine is sufficiently immunogenic and safe to initiate clinical studies with leukemia patients. PMID- 6298699 TI - [Sequential changes of plasma cyclic AMP and serum histamine following administration of the contrast material]. PMID- 6298700 TI - [First human case of Drechslera longirostrata mycosis. Spondylodiscitis complicating prosthesis endocarditis. Treatment with combined ketoconazole and amphotericin B]. AB - In a patient with spondylodiscitis secondary to cardiac valve prosthesis infection with endocarditis the fungus Drechslera longirostrata, which had not yet been known to cause mycoses, was isolated from cultures of prosthetic material and an intervertebral disc. The cardiac prosthesis had to be replaced and the vertebral lesion, which extended along 3 lumbar segments and was destructive enough to produce neurological disorders, required surgical immobilization of the spine. The disc infection was cured after combined administration of amphotericin B and ketoconazole, both drugs having proved unsuccessful when given alone. Infections caused by rare opportunistic fungi are becoming increasingly common and are difficult to diagnose since immunological methods are inapplicable. In some resistant or extremely severe fungal infections antifungal agents can be used in combinations for their synergistic effects, with subsequent reduction of dosage and potential side-effects. Combinations must be based on in vitro sensitivity tests. PMID- 6298701 TI - [Relation between monoclonal gammopathy and hepatobiliary disease]. AB - Eleven cases of monoclonal gammopathy associated with chronic hepato-biliary diseases are reported. The gammopathy was benign and the majority of hepato biliary diseases consisted of cirrhosis complicated or not by hepatocellular carcinoma. The small number of cases reported and the absence of distinctive clinical, biochemical and evolutive features of the gammopathy and hepato-biliary disease suggest that the association was fortuitous. PMID- 6298702 TI - The organization of the tadpole and adult alpha globin genes of Xenopus laevis. AB - Adult erythrocytes of X. laevis contain six electrophoretically resolvable globin polypeptides while tadpole erythrocytes contain four polypeptides, none of which comigrates with an adult protein. We show that three of the adult proteins are alpha globin polypeptides (alpha 1, alpha 2, alpha 3) and three are beta globin polypeptides (beta 1, beta 2, beta 3). We find that a tadpole alpha globin gene (alpha T1) is linked to the major adult locus in the sequence 5'-alpha T1-alpha 1 beta 1-3' with 5.2 kb separating alpha T1 from alpha 1. Another tadpole alpha globin gene (alpha T2) is linked to the minor adult locus in the sequence 5' alpha T2-alpha 2-beta 2-3' with 10.7 kb separating alpha T2 from alpha 2. These linkage relationships are consistent with the major and minor loci having arisen by tetraploidization but the different separation of larval and adult globin genes at the two loci indicates the occurrence of some additional chromosomal rearrangement. Two alternative models are presented. PMID- 6298703 TI - Transcriptional 'enhancers' from SV40 and polyoma virus show a cell type preference. AB - Transcriptional "enhancers" have been identified in both the monkey virus SV40 and in the mouse polyoma virus. Here we report that these enhancers show a cell type preference. This was done, (i) by assaying for T antigen expression and viral DNA replication of polyoma DNA which contains its own enhancer or the enhancer of SV40 virus, and (ii) by linking either of the two enhancer elements to the rabbit beta 1-globin gene and measuring transient globin expression in mouse and primate cells. The results were consistent in all the assays: In mouse cells the polyoma enhancer is slightly more effective than the SV40 enhancer. However, in primate cells the SV40 enhancer induces a four to six fold higher level of gene expression than does the polyoma enhancer. PMID- 6298704 TI - Chromatin structure of histone genes in sea urchin sperms and embryos. AB - The nucleosomal organization of active and repressed alpha subtype histone genes has been investigated by micrococcal nuclease digestion of P. lividus sperm, 32 64 cell embryo and mesenchyme blastula nuclei, followed by hybridization with 32P labeled specific DNA probes. In sperms, fully repressed histone genes are regularly folded in nucleosomes, and exhibit a greater resistance to micrococcal nuclease cleavage than bulk chromatin. In contrast, both coding and spacer alpha subtype histone DNA sequences acquire an altered conformation in nuclei from early cleavage stage embryos, i.e., when these genes are maximally expressed. Switching off of the alpha subtype histone genes, in mesenchyme blastulae, restores the typical nucleosomal organization on this chromatin region. As probed by hybridization to D.melanogaster actin cDNA, actin genes retain a regular nucleosomal structure in all the investigated stages. PMID- 6298705 TI - A nonanucleotide sequence involved in promotion of ribosomal RNA synthesis and RNA priming of DNA replication in yeast mitochondria. AB - We have examined the initiation of transcription of the mitochondrial genes for ribosomal RNA (rRNA) in the yeast Kluyveromyces lactis and show that these are transcribed independently from individual promoters. The mature large rRNA contains a 5' di- or triphosphate end which can be labelled in vitro with [alpha 32P]GTP using guanylyltransferase and this enabled us to determine the nucleotide sequence of its 5' terminus. For the small rRNA, a minor in vitro capped RNA species hybridizes in the region where--as judged from S1 nuclease protection experiments--the precursor of this RNA starts. We have determined the DNA sequence around the beginning of both rRNA genes and this reveals the existence of an identical nonanucleotide sequence (5' -ATATAAGTA- 3') just preceding the positions where the rRNAs start. This sequence is identical to the one preceding the rRNA genes in the mtDNA of the distantly related yeast Saccharomyces cerevisiae (Osinga, K.A. and Tabak, H.F. (1982) Nucl.Acids Res. 10, 3617-3626) and supports our proposal that this sequence motif is part of a yeast mitochondrial promoter. We have noticed that the same sequence is located in the putative origin of replication present in hypersuppressive petite mutants of S. cerevisiae and consider the possibility that this sequence is involved in RNA priming of DNA replication. PMID- 6298706 TI - Nucleotide sequence at the site of junction between adenovirus type 12 DNA and repetitive hamster cell DNA in transformed cell line CLAC1. AB - The hamster cell line CLAC1 originated from a tumor induced by injecting human adenovirus type 12 (Ad12) into newborn hamsters. Each cell contained about 12 copies of viral DNA colinearly integrated at two or three different sites. We have cloned and sequenced a DNA fragment comprising the site of junction between the left terminus of Ad12 DNA and cellular DNA. The first 174 nucleotides of Ad12 DNA were deleted at the site of junction. Within 40 nucleotides, there were one tri-, two tetra-, one penta-, and one heptanucleotide which were identical in the 174 deleted viral nucleotides and the cellular sequence replacing them. In addition, there were patch-type homologies ranging from octa- to decanucleotides between viral and cellular sequences. There is no evidence for a model assuming adenovirus DNA to integrate at identical cellular sites. The cellular DNA sequence corresponding to the junction fragment was cloned also from BHK21 (B3) hamster cells and sequenced. Up to the site of linkage with viral DNA, this middle repetitive cellular DNA sequence was almost identical with the equivalent sequence from CLAC1 hamster cells. Taken together with the results of previously published analyses (11, 12), the data suggest a model of viral (foreign) DNA integration by multiple short sequence homologies. Multiple sets of short patch homologies might be recognized as patterns in independent integration events. The model also accounts for the loss of terminal viral DNA sequences. PMID- 6298707 TI - Sequence of rat alpha- and gamma-casein mRNAs: evolutionary comparison of the calcium-dependent rat casein multigene family. AB - The complete sequences of rat alpha- and gamma-casein mRNAs have been determined. The 1402-nucleotide alpha- and 864-nucleotide gamma-casein mRNAs both encode 15 amino acid signal peptides and mature proteins of 269 and 164 residues, respectively. Considerable homology between the 5' non-coding regions, and the regions encoding the signal peptides and the phosphorylation sites, in these mRNAs as compared to several other rodent casein mRNAs, was observed. Significant homology was also detected between rat alpha- and bovine alpha s1-casein. Comparison of the rodent and bovine sequences suggests that the caseins evolved at about the time of the appearance of the primitive mammals. This may have occurred by intragenic duplication of a nucleotide sequence encoding a primitive phosphorylation site, -(Ser)n-Glu-Glu-, and intergenic duplication resulting in the small casein multigene family. A unique feature of the rat alpha-casein sequence is an insertion in the coding region containing 10 repeated elements of 18 nucleotides each. This insertion appears to have occurred 7-12 million years ago, just prior to the divergence of rat and mouse. PMID- 6298708 TI - Sequence of the junction in adenovirus 2-SV40 hybrids: examples of illegitimate recombination. AB - The nucleotide sequences of six Ad2-SV40 junctions from three Ad2-SV40 hybrid viruses (Ad2++HEY, Ad2++LEY and Ad2+D1) were determined. Comparison of parental adenovirus 2 and SV40 DNA sequences with the sequence at the Ad2-SV40 junctions revealed that 5 out of 6 junctions are abrupt transitions from Ad2 to SV40 DNA, and in one case (Ad2++LEY, right junction) there is an additional nucleotide at the junction, which cannot be ascribed to either DNA. Ad2++HEY and Ad2+D1 right junctions are identical and Ad2++LEY and Ad2+ND4 left junctions are identical, a result that strongly suggests these Ad2-SV40 hybrids arose by recombination between the linear Ad2 DNA and circular SV40 DNA, followed by recombination between Ad2 DNA and SV40 DNA present in the Ad2-SV40 hybrid DNA. The unambiguous transition of Ad2 DNA into SV40 DNA at the junction sites is an example of recombination events which have apparently occurred without any homology at the recombination site. PMID- 6298709 TI - Cloning and expression of the cDNA for human antithrombin III. AB - A partial cDNA clone for human antithrombin III (ATIII) was obtained by screening a cDNA library prepared from size fractionated liver RNA with a pool of eight 16 base long synthetic DNA fragments whose sequence was determined from protein sequence data. A fragment of the partial cDNA clone was used to enrich RNA for ATIII messages, and cDNA clones encoding the entire ATIII structural gene were identified. The complete nucleotide and predicted amino acid sequences of human ATIII and its 32 residue signal peptide are reported, and provide further opportunity to compare the ATIII primary structure with corresponding regions from homologous proteins, alpha 1-antitrypsin and ovalbumin. Plasmids in which the structural genes for mature and pre-ATIII were linked to the E. coli trp promoter-operator support the synthesis of human antithrombin III and pre antithrombin III in bacteria. PMID- 6298711 TI - ScrFI: a new sequence-specific endonuclease from Streptococcus cremoris. AB - A novel sequence-specific endonuclease has been isolated from Streptococcus cremoris F. ScrFI recognises the sequence: (formula; see text) and cleaves as indicated by the arrow ( ). It is the first enzyme to recognise this sequence and the first endonuclease reported from the lactic streptococci used in dairy fermentations. PMID- 6298710 TI - An in vitro RNA polymerase III system from S. cerevisiae: effects of deletions and point mutations upon SUP4 gene transcription. AB - A soluble cell-free extract containing RNA polymerase III and factors essential for selective transcription of the yeast SUP4-o tRNATyr gene was prepared from Saccharomyces cerevisiae cells. An intragenic promoter for yeast RNA polymerase III was identified within the yeast tRNATyr coding sequence by testing several sup4 genes with 5'- and 3'-terminal deletions in the homologous transcription system. Thirty-four different sup4 genes with spontaneous mutations were also tested in the in vitro system. Two point mutations drastically reduced transcription initiation and two other mutations caused premature termination. These mutations have nearly identical effects on SUP4 gene transcription by Xenopus RNA polymerase III (1), which demonstrates that the essential features of RNA polymerase III transcription initiation and termination signals have been conserved throughout the course of eukaryotic evolution. PMID- 6298712 TI - Synthetic deoxyoligonucleotides as general probes for chloroplast transfer RNA genes. AB - The utility of chemically synthesized deoxyoligonucleotides as hybridization probes for the detection of tRNA genes has been examined. Chloroplast tRNA genes were chosen for this study. Deoxyoligonucleotides complementary to highly conserved regions of chloroplast tRNA genes of both higher plants and Euglena gracilis were chemically synthesized. These synthetic probes have been used to detect tRNA genes by Southern hybridizations to restriction fragments of chloroplast DNAs. This new method of tRNA gene mapping and the oligonucleotides synthesized may be of general application to many chloroplast genomes. This is illustrated by the detection of known and unknown tRNA genes of Euglena gracilis and spinach, and unknown tRNA genes of maize and cucumber chloroplast DNAs. The precise locus and polarity of the Euglena gracilis chloroplast tRNAPhe gene has been determined. We also describe experiments which relate to the effects of the time of hybridization, the stringency of washing, and of base pair mismatches on the hybridization signal. PMID- 6298713 TI - Closely related families of genes code for the alpha and alpha' subunits of the soybean 7S storage protein complex. AB - Nineteen cloned cDNAs encoding the alpha and alpha'-subunits of the 7S seed storage protein in the soybean, Glycine max, have been isolated from a recombinant cDNA library constructed with mRNA from maturing seeds. In addition, a gene encoding an alpha'-subunit has been isolated from a recombinant Charon 4A phage library containing genomic Glycine max DNA. The cloned DNAs have been divided, on the basis of their endonuclease sites, into two main classes of sequences which differ in approximately 6% of their nucleotides. Whereas the proteins encoded within each DNA class are nearly identical, the proteins encoded by the two different classes of soybean DNAs are distinct and correspond to alpha and alpha'-subunits. Thus, the alpha and alpha'-subunits are coded for by two closely related multigene families. The amino acid differences in the portions of the alpha and alpha'-subunits presented in this paper occur primarily near the carboxyl-terminus. The 3' noncoding nucleotides of the cloned alpha and alpha' subunit DNAs are more highly conserved than are the coding nucleotides. This conservation suggests that the 3' untranslated sequences of the alpha and alpha' subunit mRNAs are functional in the expression of the alpha and alpha'-subunit proteins or in the stabilization of the 7S subunit mRNAs. PMID- 6298714 TI - Sequence organization of the ribosomal spacer of D.melanogaster. AB - We subcloned several fragments of the rDNA nontranscribed spacer (NTS) of D.melanogaster and analysed their nucleotide sequence. The central portion contains a variable number of 240 base pair repeated sequences ending with two contiguous Alu I sites. Within every repeat there is a 42 base pair sequence perfectly homologous to the transcription initiation site from -18 to +24. Sequences homologous to the basic 240 bp repeat are present both at the end of the unique 1900 bp region downstream to the transcription termination site and within the unique region preceding the transcription initiation site up to nucleotide -140. These results are discussed in relation to the general problem of ribosomal gene units and their evolution. PMID- 6298715 TI - Comparison of the amino acid sequence of the major immunogen from three serotypes of foot and mouth disease virus. AB - Cloned cDNA molecules from three serotypes of FMDV have been sequenced around the VP1-coding region. The predicted amino acid sequences for VP1 were compared with the published sequences and variable regions identified. The amino acid sequences were also analysed for hydrophilic regions. Two of the variable regions, numbered 129-160 and 193-204 overlapped hydrophilic regions, and were therefore identified as potentially immunogenic. These regions overlap regions shown by others to be immunogenic. PMID- 6298716 TI - A comparison of nucleoside (beta-S)triphosphates and nucleoside (gamma S)triphosphates as suitable substrates for measuring transcription initiation in preparations of cell nuclei. AB - RNA chains initiated with nucleoside (beta-S)triphosphates and (gamma S)triphosphates retain the thiol groups and can be separated from thiol-free RNA by chromatography on mercury-Sepharose. Thiol-containing mouse mammary tumor virus (MMTV) RNA synthesized by preparations of nuclei from virus-infected cells was quantitated by nucleic acid filter hybridization. With ATP beta S and GTP beta S, region-specific initiation of MMTV RNA chains was detected in the cell free system. However, with ATP gamma S and GTP gamma S, region-specific initiation was not clearly demonstrable. The nuclear preparations can also transfer thiol groups, presumably in the form of thiophosphate, from ATP gamma S or GTP gamma S onto preexisting RNA molecules; little or no thiol-transfer occurs with the two (beta-S)-analogues. The thiophosphate transfer activity apparently interferes with the measurement of RNA chain initiation with ATP gamma S and GTP gamma S. PMID- 6298717 TI - A small DNA molecule containing covalently-linked ribonucleotides originates from the large intergenic region of the cauliflower mosaic virus genome. AB - We have detected a small DNA molecule (sa-DNA), 725 nucleotides long, in cauliflower mosaic virus (CaMV)-infected, but not non-infected, turnip leaves. Alkali and RNase A treatments shortened sa-DNA by 100 nucleotides and we conclude that it contains covalently-linked ribonucleotides. This DNA co-purified with cellular polyadenylated RNA. It is complementary to the beta-strand of CaMV DNA and of opposite polarity to RNAs transcribed from the alpha-strand. Hybridisation studies suggest that sa-DNA originates from the large intergenic region (IR1) of the CaMV genome. A small double-stranded DNA with three single-stranded components, which co-purifies with cellular DNA, appears to be related to sa-DNA but lacks detectable ribonucleotides. PMID- 6298718 TI - DNA methylation patterns in the 5S DNAs of Xenopus laevis. AB - The frequency of cytosine methylation at specific sites in the somatic 5S DNA (X1s) and trace oocyte 5S DNA (X1t) of X. laevis has been determined using restriction enzymes that are inhibited by the presence of 5-methylcytosine (5mC) within their cleavage sequences. 5S DNA methylation patterns were determined in genomic DNA from mature red blood cells, which express neither type of 5S gene, and from liver, which expresses only X1s. All the sites examined in X1t are greater than 95% methylated in red cells and liver. In the X1s of red cells all the sites examined are methylated in greater than 95% of repeats, while in liver some sites are modified in only 90% of repeats. Repeats containing unmethylated sites are randomly distributed throughout the tandem arrays in both red cells and liver. The high levels of methylation for X1s are in marked contrast to the situation with other Xenopus genes which do have sites of significant undermethylation in tissues where they are active. Thus, undermethylation in active genetic regions may not be a general feature for all classes of eukaryotic genes. PMID- 6298719 TI - The independent evolution of two closely related satellite DNA elements in rats (Rattus). AB - We have identified and determined the sequence and organization of a new rat satellite DNA in Rattus rattus, the roof rat. This satellite DNA, which we call R. rattus satellite I', consists of tandem arrays of a 185 base pair (bp) repeat unit that we call a'. a' is 86% homologous to a 185 bp portion of the 370 bp repeat unit of the previously described rat satellite [Pech et al. (1979) Nucleic Acids Res. 7, 417-432] present in the common laboratory rat species, R. norvegicus. We can thereby distinguish two 185 bp portions of the satellite I 370 bp repeat unit: "a" (homologous to a') and "b". Satellite I has the structure (a,b)n, and satellite I' has the structure (a')n. Like a, a' is only about 60% homologous to b and fails to hybridize to b. Although R. norvegicus and R. rattus contain about the same total concentration of satellite sequences, R. norvegicus contains essentially only the a,b type (satellite I), whereas R. rattus contains a' type (satellite I') and lesser amounts of the a,b type (satellite I). The a,b type (satellite I) in R. rattus is very similar to that in R. norvegicus as judged both by hybridization and by the presence of all but one of the major restriction enzyme sites that characterize the R. norvegicus satellite I. In R. rattus the a' and a,b repeat units are not detectably present in the same tandem array. All of the sequence differences between a' (R. rattus) and a (R. norvegicus) can be accounted for by simple base substitutions, and the implication of this and other features of rat satellite DNA structure for satellite DNA evolution are discussed. PMID- 6298720 TI - Discrete regions of sequence homology between cloned rodent VL30 genetic elements and AKV-related MuLV provirus genomes. AB - Southern blot analyses using reduced stringency hybridization conditions have been employed to search for sequence homologies between rodent VL30 genes and murine leukemia virus (MuLV) proviruses. These constitute two classes of transposon-like elements previously believed to be genetically unrelated. Our results demonstrate that cloned representatives of both ecotropic and xenotropic like proviruses share discrete regions of sequence homology with VL30 genes of both rat and mouse origin. These regions of homology exist in both 3' and 5' halves of the MuLV genome but do not include extensive portions of the long terminal repeat (LTR) or a 0.4 Kbp segment of the env gene specific for recently acquired ecotropic-type MuLV proviruses. DNA sequencing, however, revealed that the short inverted terminal repeat sequence of MuLV proviral LTRs is almost perfectly conserved at the terminus of an integrated mouse VL30 gene. These results suggest that recombination events with rodent VL30-type sequences occurred during early MuLV evolution. The strong conservation of the inverted terminal repeat sequence may reflect a common integration mechanism for VL30 elements and MuLV proviruses. PMID- 6298721 TI - Members of the KpnI family of long interspersed repeated sequences join and interrupt alpha-satellite in the monkey genome. AB - Three different members of a family (KpnI-family) of interspersed repeated DNA sequences were found linked to alpha-satellite sequences in cloned segments of the African green monkey genome. In two of these segments the KpnI-family member is over 6 kbp in length and one of them is flanked by alpha-satellite on both sides indicating that it was inserted into a satellite array. Hybridization of subcloned portions of the family members to restriction endonuclease digests of monkey and human DNA and to a genomic library of African green monkey DNA indicate that 1) family members are interspersed in both the monkey and human genomes, 2) some family members may include sequences in addition to those in the three characterized here, 3) some family members may contain only parts of the sequences characterized here and 4) while the overall organization of the family is similar in the human and monkey genome the majority of the family members in each of the two genomes are distinctly identified by the variant position of certain restriction endonuclease sites. This last observation suggests that within each genome there is a tendency to maintain particular versions of the sequence. Observations 2) and 3) suggest that the KpnI family is complex and includes a variety of subfamilies. PMID- 6298722 TI - Complete DNA sequence coding for the large ribosomal RNA of yeast mitochondria. AB - The mitochondrial gene coding for the large ribosomal RNA (21S) has been isolated from a rho- clone of Saccharomyces cerevisiae. A DNA segment of about 5500 base pairs has been sequenced which included the totality of the sequence coding for the mature ribosomal RNA and the intron. The mature RNA sequence corresponds to a length of 3273 nucleotides. Despite the very low guanine-cytosine content (20.5%), many stretches of sequence are homologous to the corresponding Escherichia coli 23S ribosomal RNA. The sequence can be folded into a secondary structure according to the general models for prokaryotic and eukaryotic large ribosomal RNAs. Like the E.coli gene, the mitochondrial gene contains the sequences that look like the eukaryotic 5.8S and the chloroplastic 4.5S ribosomal RNAs. The 5' and 3' end regions show a complementarity over fourteen nucleotides. PMID- 6298723 TI - Kilo-sequencing: an ordered strategy for rapid DNA sequence data acquisition. AB - A strategy for rapid DNA sequence acquisition in an ordered, nonrandom manner, while retaining all of the conveniences of the dideoxy method with M13 transducing phage DNA template, is described. Target DNA 3 to 14 kb in size can be stably carried by our M13 vectors. Suitable targets are stretches of DNA which lack an enzyme recognition site which is unique on our cloning vectors and adjacent to the sequencing primer; current sites that are so useful when lacking are Pst, Xba, HindIII, BglII, EcoRI. By an in vitro procedure, we cut RF DNA once randomly and once specifically, to create thousands of deletions which start at the unique restriction site adjacent to the dideoxy sequencing primer and extend various distances across the target DNA. Phage carrying a desired size of deletions, whose DNA as template will give rise to DNA sequence data in a desired location along the target DNA, may be purified by electrophoresis alive on agarose gels. Phage running in the same location on the agarose gel thus conveniently give rise to nucleotide sequence data from the same kilobase of target DNA. PMID- 6298724 TI - Structure and transcription of the nopaline synthase gene region of T-DNA. AB - We present the DNA sequence and plant-tumor transcription pattern of some 2400 base pairs from the right border region of pTi T37 DNA from the virulent Agrobacterium tumefaciens strain T37. This region includes the entire transcription unit encompassing the nopaline synthase gene, together with parts of other transcription units. The strategy used to determine the sequence also produced two opposing series of defined, asymmetric deletions across the target DNA region, some of which may serve future purposes in the exploitation of this sequence, which is known to be expressed in a wide variety of host plant tissues. PMID- 6298725 TI - Affinity of HMG17 for a mononucleosome is not influenced by the presence of ubiquitin-H2A semihistone but strongly depends on DNA fragment size. AB - We have used a two-dimensional (deoxyribonucleoprotein leads to DNA) electrophoretic binding assay to study the interaction of the purified high mobility group protein HMG17 with isolated HeLa mononucleosomes as a function of their DNA fragment size and the presence of ubiquitin-H2A semihistone. No significant differences between affinities of HMG17 for ubiquitinated and non ubiquitinated core mononucleosomes were observed. In striking contrast, the apparent affinity of HMG17 for a mononucleosome increases more than 100-fold upon an increase of the length of the mononucleosomal DNA fragment by as few as 3 to 5 bp over the core DNA length (integral of 146 bp). We suggest that the magnitude of this effect is sufficient to explain the preferential binding of HMG17 in vitro to mononucleosomes derived from actively transcribed genes. PMID- 6298726 TI - Association of eukaryotic DNA topoisomerase I with nucleosomes and chromosomal proteins. AB - A DNA topoisomerase activity is found to be associated with the nucleosomes released by the Staphylococcal nuclease digestion of HeLa nuclei. Such an association is found to be salt dependent. A number of criteria have established that this DNA topoisomerase activity is due to HeLa topo I (Liu, L. F. and Miller, K. G. (1980) Proc. Natl. Acad. Sci. USA 78, 3489-3491). A similar association has been demonstrated from the in vitro studies using purified mononucleosomes and eukaryotic DNA topoisomerase I. Nonhistone HMG proteins and histone H1 are found to stimulate topoisomerase activity in vitro and form tight complexes with eukaryotic DNA topoisomerase I. The intimate interactions of topoisomerase I with chromosomal proteins and nucleosomes may be an essential feature of the topoisomerase function in vivo. PMID- 6298727 TI - Does the specific recognition of DNA by the restriction endonuclease EcoRI involve a linear diffusion step? Investigation of the processivity of the EcoRI endonuclease. AB - The time course of the EcoRI endonuclease catalysed cleavage of three substrates, two plasmid DNAs and one oligonucleotide, each with two EcoRI sites, was measured. The two plasmid DNAs with the EcoRI sites 318 and 96 base pairs apart are cut in a distributive fashion, while the oligonucleotide with the EcoRI sites 8 base pairs apart is cut in a partially processive manner. It is concluded that a linear diffusion of the EcoRI endonuclease on its substrate across long stretches of DNA is not likely to be operative during the recognition process. Microscopic dissociation-reassociation processes, however, increase the probability of the enzyme to attack further sites located in the immediate vicinity of a given site. PMID- 6298728 TI - Nucleotide sequence of the Tn10 encoded tetracycline resistance gene. AB - The nucleotide sequence of 1530 base pairs of Tn10 DNA coding for tetracycline resistance has been determined. The gene start consists of overlapping bidirectional promotors and two operator sequences. One terminator of transcription as defined by the typical terminator sequence is about 1300 base pairs downstream from the promotor. It is preceeded by translation termination codons in all three possible reading frames. The transcript contains an open reading frame coding for a 43.3 kDa protein. Two other possible reading frames are discussed. The amino acid sequence of the Tn10 encoded tetracycline resistance gene shows good homology with two proteins encoded in the tetracycline resistance part of the plasmid pBR322. The hydrophobic nature of the 43.3 kDa protein is discussed with regard to it's proposed function. PMID- 6298729 TI - Polyoma-induced stimulation of nucleoplasmic transcription is paralleled by development of resistance against actinomycin D. AB - Polyoma virus induced in quiescent, Go-arrested mouse kidney cells a lytic infection. Synthesis of the polyoma T-antigens began 7-8 h after infection and was followed by a mitotic reaction of the host cell comprising stimulated synthesis and accumulation of cellular (mainly ribosomal) RNA and protein and duplication of the host cell chromatin (S-phase). In the present work we focused attention on nucleoplasmic transcription, i.e. synthesis of hnRNA, 5S RNA and tRNA. To inhibit selectively nucleolar transcription we used low concentrations of actinomycin D (act. D). Synthesis of 45S precursor- ribosomal RNA in mock- and polyoma-infected mouse kidney cells was completely blocked by 0.05 micrograms/ml act.D within 2 h. In mock-infected cells also nucleoplasmic transcription was rather sensitive against 0.05 micrograms/ml act.D. Polyoma- induced stimulation of nucleoplasmic transcription began around 12 h and was paralleled by the development of resistance against act.D. Resistance of nucleoplasmic transcription in virus-infected cells was thus similar to that observed by others in uninfected, proliferating mammalian cells. The possible biological implications of these results are discussed. PMID- 6298730 TI - DNA topoisomerases from rat liver: physiological variations. AB - Besides the nicking-closing (topoisomerase I) activity, an ATP-dependent DNA topoisomerase is present in rat liver nuclei. The enzyme, partially purified, is able to catenate in vitro closed DNA circles in a magnesium-dependent, ATP dependent, histone H1-dependent reaction, and to decatenate in vitro kinetoplast DNA networks to yield free minicircles in a magnesium-dependent and ATP-dependent reaction. It is largely similar to other eukaryotic type II topoisomerases in its requirements, and presumably belongs to this class of enzymes. Type I and type II activities were measured in rat liver nuclei as a function of regenerating time after partial hepatectomy: type I activity was not significantly changed during this process. In contrast, type II activity was considerably increased, suggesting a possible involvement of the enzyme in DNA replication. PMID- 6298732 TI - Isolation and nucleotide sequence of a plant tRNA gene: petunia asparagine tRNA. AB - A 14.3 kb petunia genomic DNA fragment was isolated and found to contain a single tRNA gene coding for asparagine tRNA. The nucleotide sequence of the asparagine tRNA gene and its flanking regions has been determined. This gene does not contain intervening sequences nor the 3'-end CCA sequence of the mature tRNA and presents a similar overall sequence homology (70%) to both E. coli and mammalian asparagine tRNA. As in other eukaryotic tRNA genes the 5'-flanking region does not seem to contain any special sequence that could function as a regulatory element and the 3'-end is followed by a short cluster of T that may function as the transcription termination site. PMID- 6298731 TI - The coupled use of 'footprinting' and exonuclease III methodology for RNA polymerase binding and initiation. Application for the analysis of three tandem promoters at the control region of colicin El. AB - In order to determine the initiation site for three promoters P1, P2 and P3 (5' to 3') in close proximity in the colicin E1 control region we developed a new methodology that couples ternary complex formation and the analysis of the 3' border protected from exonuclease III digestion. The initiation of transcription could be detected by measuring the shift in the position of the 3' protected border when RNA polymerase moved from its binary complex position to its ternary complex position. The latter stops at a specific nucleotide because transcription is initiated with one or more NTPs missing. This approach, coupled with "footprinting", can also be used to decide whether the formation of an RNA polymerase binary or ternary complex at one site excludes or weakens binding at neighboring sites. The location of 3' protected borders reveals the formation of respective binary and ternary complexes at non-saturating RNA polymerase conditions, whereas at saturating conditions only the distal 3' boundary is seen and exonuclease cannot penetrate further. However, if "footprinting" reveals proximal 5' patterns this establishes that simultaneous binding has occurred on the same DNA fragment. The data showed that this was true for P1 and P3 which are only 8 nucleotides apart. P2 could only be detected at non-saturating conditions since it overlaps both P1 and P3. The evidence from the literature and this study establishes P1 as the true colicin E1 promoter with the possibility that supercoiling may eliminate any role for P2 and P3. PMID- 6298733 TI - The nucleotide sequence of myosin light chain (L-2A) mRNA from embryonic chicken cardiac muscle tissue. AB - The nucleotide sequence of a cDNA clone (pML10) for chicken cardiac myosin light chain is described. The cDNA insert contains 613 nucleotides representing the entire coding sequence, with the exception of nine NH2-terminal amino acids, and the full 3'-non-coding region of 146 nucleotides. The missing 5' terminus of the mRNA, not represented in the clone pML10, was obtained by extension of the cDNA using a 43 nucleotide long internal EcoR1 fragment as a primer. The non-coding region contains several direct and inverted repeated sequences and the polyadenylation signal sequence AATAAA. The coding portion exhibits non-random usage of synonymous codons with a strong bias for codons ending in G and C. PMID- 6298735 TI - Histone gene number and organisation in Xenopus: Xenopus borealis has a homogeneous major cluster. AB - Using a Xenopus laevis H4 cDNA clone as a probe we have determined that the numbers of H4 histone genes in Xenopus laevis and Xenopus borealis are approximately the same. These numbers are dependent on the hybridization stringency and we measure about 90 H4 genes per haploid genome after a 60 degrees C wash in 3 X SSC. Using histone probes from both Xenopus and sea urchin we have studied the genomic organization of histone genes in these two species. In all of the X.borealis individuals analyzed about 70% of the histone genes were present in a very homogeneous major cluster. These genes are present in the order H1, H2B, H2A, H4 and H3, and the minimum length of the repeated unit is 16kb. In contrast, the histone gene clusters in X.laevis showed considerable sequence variation. However two major cluster types with different gene orders seem to be present in most individuals. The differences in histone gene organization seen in species of Xenopus suggest that even in closely related vertebrates the major histone gene clusters are quite fluid structures in evolutionary terms. PMID- 6298734 TI - Isolation and characterisation of genes for androgen-responsive secretory proteins of rat seminal vesicles. AB - Under the influence of testosterone, rat seminal vesicles synthesise large amounts of a tissue specific protein, S. Recombinant lambda clones have been isolated containing overlapping sequences covering a 27.5 kilo base region of the rat genome within which the gene for protein S is located. Recombinant plasmids bearing cDNA sequences for protein S were constructed in pBR328. One (pcS2) contains a 690 nucleotide insert and is probably full length. Detailed restriction maps of the S-gene are presented and the structure was confirmed by analysis of R-loops and heteroduplexes. The S-gene covers a 2 kbp region of the genome and consists of a 5' intron (490 bp) separating a leading exon (120 bp) containing the 5' untranslated region from a central exon (310 bp) containing most of the coding sequence and part of the 3' untranslated region. A larger intron (1100 bp) lies within the 3' untranslated region. The cloned gene is representative of the native gene but the S gene may be heterogeneous. Using pcS2, the hormonal control of S-specific mRNA was examined and a pronounced differential response to testosterone was observed. PMID- 6298736 TI - The dnaC protein of Escherichia coli. Purification, physical properties and interaction with dnaB protein. AB - E.coli dnaC protein was purified to near-homogeneity in using a dnaC complementation assay [S. Wickner, I.Berkower, M.Wright, and J. Hurwitz (1973) Proc. Natl. Acad. Sci. USA 70, 2369-2373]. Purification was achieved by taking advantage of the hydrophobic interaction of dnaC protein with aliphatic and aromatic matrixes and with Brij58 as stabilizing agent. A sedimentation coefficient for the dnaC protein of 2.6 S corresponding to a molecular weight of approximately 26,000 was estimated from glycerol gradient centrifugation. A polypeptide molecular weight of 28,000 was determined by densitometry on a denaturing gel. In the presence of ATP the dnaC protein forms a complex with dnaB protein [S. Wickner and J. Hurwitz (1975) Proc.Natl.Acad.Sci. USA 72, 921-925]. For the dnaB . dnaC complex a sedimentation coefficient of 14.5 S was measured by glycerol gradient centrifugation, indicating a molecular weight of about 400,000. The ratio of the dnaC and dnaB polypeptides in the complex is approximately 1, as determined on a denaturing gel. It is suggested that the complex consists of the dnaB protein hexamer and six dnaC polypeptides amounting to a calculated molecular weight of about 450,000. PMID- 6298737 TI - The complete DNA sequence of minute virus of mice, an autonomous parvovirus. AB - We have determined the complete nucleotide sequence of the genome of Minute Virus of Mice, an autonomous parvovirus. This single-stranded DNA is 5081 nucleotides long. The 3'-and 5'-ends of the viral strand contain imperfect palindromic sequences which consist, respectively, of 115 and 206 nucleotides. The 3' terminal palindrome is composed of a unique sequence, whereas the 5'-terminal palindrome contains two sequences in equimolar amounts; these are related in that one is the inverted complement of the other. The DNA strand complementary to that which is encapsidated into virions contains two large open reading frames which together span almost the entire genome. Transcriptional and translational signals within the sequence have been identified and related to the known map coordinates of the viral transcripts. In this report we summarize some of the salient structural and organizational features of the MVM genome. PMID- 6298738 TI - Complete nucleotide sequence of alfalfa mosaic virus RNA 1. AB - Double-stranded cDNA of alfalfa mosaic virus (AlMV) RNA 1 has been cloned and sequenced. From clones with overlapping inserts, and other sequence data, the complete primary sequence of the 3644 nucleotides of RNA 1 was deduced: a long open reading frame for a protein of Mr 125,685 is flanked by a 5'-terminal sequence of 100 nucleotides and a 3' noncoding region of 163 nucleotides, including the sequence of 145 nucleotides the three genomic RNAs of AlMV have in common. The two UGA-termination codons halfway RNA 1, that were postulated by Van Tol et al. (FEBS Lett. 118, 67-71, 1980) to account for partial translation of RNA 1 in vitro into Mr 58,000 and Mr 62,000 proteins, were not found in the reading frame of the Mr 125,685 protein. PMID- 6298739 TI - Poliovirus type 3: molecular cloning of the genome and nucleotide sequence of the region encoding the protease and polymerase proteins. AB - Overlapping cDNA clones representing the entire genome of poliovirus type 3 have been prepared in E. coli by two separate methods. Cloning of RNA . cDNA hybrids produced a more comprehensive set of clones with generally larger cDNA inserts than cloning of double - stranded cDNA. A restriction map of the entire genome and the nucleotide sequence of 2003 bases from the 3' terminus, comprising the region encoding the protease and polymerase proteins, are presented. PMID- 6298740 TI - Expression of chemically synthesized alpha-neo-endorphin gene fused to E. coli alkaline phosphatase. AB - An alpha-neo-endorphin (alpha NE) gene, which we previously synthesized chemically and inserted into E. coli beta-galactosidase gene of pK013 plasmid, has been excised and fused to E. coli alkaline phosphatase (APase) gene. One of the transformants was named E15/pA alpha NE1. Under the APase gene regulation, APase-alpha NE chimeric protein was expressed at 1.3 X 10(6) molecules per cell, and accounted for about 60% of total cellular proteins. The HPLC pattern of CNBr treated E15/pA alpha NE1 was very simple reflecting the high content of the chimeric protein and low numbers of methionine residues in it. A series of genes encoding APase-alpha NE chimeric proteins in which 30 to 94 C-terminal amino acid residues were replaced by (met)-alpha NE, was cloned in E. coli. Transportation of the chimeric proteins to periplasmic space was studied. All chimeric proteins were apparently processed by signal peptidase but few, if any, was transported to the periplasmic space. PMID- 6298741 TI - High efficiency polyoma DNA transfection of chloroquine treated cells. AB - Chloroquine treatment of rodent cells during the first hours of polyoma DNA transfection increase the fraction of cells expressing viral functions. The effect has been observed after DNA absorption using both the DEAE-dextran and calcium phosphate coprecipitation methods. Exposure to chloroquine increased the proportion of transfected mouse cells to approximately 40%. From a culture of one million such cells, microgram quantities of newly synthesized viral DNA could be isolated. Similarly, the transformation frequency of rat cells following polyoma DNA transfection was approximately 6-fold increased by chloroquine treatment. The effect of the compound was even more pronounced in transfections with linear forms of polyoma DNA, suggesting that chloroquine inhibits degradation of DNA absorbed by the cells. PMID- 6298742 TI - Comparative structure and evolution of goat and sheep satellite I DNAs. AB - The satellite I DNAs of domestic goat (Capra hircus) and domestic sheep (Ovis aries) have been studied using molecular hybridisation and restriction enzyme analysis. Both satellite DNAs are composed of repeat units of 820 base pairs in length, but their restriction maps, although similar, differ in certain respects. Thus the majority of sheep satellite I repeat units have two EcoRI sites and one AluI site, whereas the majority of goat satellite I repeat units have one EcoRI site and two AluI sites. The sheep satellite I repeat units with the two EcoRI sites are much more homogeneous than the repeats forming the remainder of the satellite, as judged by the difference in the melting temperatures of native and reassociated molecules. DNAs from species of wild sheep and goats were screened for the presence of these repeat units, and they appear to have been amplified during the radiation of the Ovis genus. Goat satellite I is composed of a single sequence type which has changed through base substitution until the sequence now shows considerable heterogeneity. It is proposed that the major sequence types of these two satellite DNAs were amplified by different saltatory replication events at different times in the evolution of the group. PMID- 6298743 TI - Methylation of the chicken vitellogenin gene: influence of estradiol administration. AB - The degree of methylation of the chicken vitellogenin gene has been investigated. Upon induction by administration of estradiol to a rooster, methyl groups at specific sites near the 5'-end of the gene are eliminated. The process of demethylation is slower than the activation of the gene. Demethylation is therefore probably not a prerequisite to gene transcription. At least two other sites in the coding region of the gene are methylated in the liver of estrogenized roosters, but not in the liver of a laying hen, where the gene is naturally active. PMID- 6298744 TI - A G43 to U43 mutation in E. coli tRNAtyrsu3+ which affects processing by RNase P. AB - A novel mutation in the anticodon stem of E. coli tRNA1Tyrsu3+ (G43 to U43) has been characterized. The gene coding for the mutant tRNA, carried by phage phi 80DHA61.3 a derivative of phi 80psu3+su0, produces only 20% of mature suppressor tRNA as compared with phi 80psu3+. Both the mutant tRNA precursor and mature tRNA have an altered conformation. The precursor tRNA coded for by phi 80DHA61.3 is processed by RNase P more slowly than the su3+ precursor and does not form as stable an enzyme-substrate complex as does su3+ precursor. phi 80 DHA61.3 also contains a large deletion which begins in the spacer region between the su3+ gene and the su0- gene, extends through the su0- gene and includes most of the repeated region following the tRNA genes. PMID- 6298745 TI - Characterization of the herpes simplex virus type 1 glycoprotein D mRNA and expression of this protein in Xenopus oocytes. AB - We have identified and characterized a 3.0 kilobase (kb) mRNA containing coding sequences of the herpes simplex virus type 1 (HSV-1) glycoprotein D (gD) gene. The synthesis of this 3.0 kb mRNA was unaffected by the presence of cytosine arabinoside, but was made in greatly reduced amounts in cells infected with HSV-1 in the presence of cycloheximide: it was, therefore, classified as an early mRNA. By nuclease protection experiments, it was found that the 3.0 kb mRNA is unspliced and, further, that it is 3' co-terminal with a smaller 1.6 kb early mRNA which is transcribed from a DNA sequence 3' to the gD coding sequence. We describe the use of the Xenopus laevis oocyte system to produce HSV-1 gD in vitro. Oocytes injected with mRNA isolated from HSV-1-infected Vero cells synthesized gD, which was identified by immunoprecipitation. Injection of a plasmid clone containing the HSV-1 BamHI J fragment (0.89 to 0.93 map units) into the nuclei of Xenopus oocytes also resulted in synthesis of gD. PMID- 6298746 TI - Characterization of a prokaryotic topoisomerase I activity in chloroplast extracts from spinach. AB - A topoisomerase I activity has been partially purified from crude extracts of spinach chloroplasts. This activity relaxes the supercoiled covalently closed circular DNA of pBR322. The enzyme requires Mg++, but not ATP, and has an apparent molecular weight of about 115,000. It catalyzes a unit change in the linkage number of supercoiled DNA but cannot relax positive supercoiled DNA. These characteristics of the topoisomerase suggest it is of the prokaryotic type and would tend to support the endosymbiotic theory of plastid origin and evolution. PMID- 6298747 TI - Complete nucleotide sequence of a cloned cDNA derived from the major adult alpha globin mRNA of X. laevis. AB - The complete sequence of a cloned cDNA derived from the major adult alpha-globin mRNA of Xenopus laevis (the South African Clawed Toad) is presented. The sequence contains the complete coding and 3' non-coding regions of the mRNA and part of the 5' non-coding region. The amino acid sequence of the encoded alpha-globin polypeptide has been deduced and is compared to other alpha-globin polypeptides. We find that the sequence is equally diverged from a bullfrog tadpole alpha globin polypeptide and human alpha-globin polypeptide suggesting that these three sequences may have diverged from a common ancestral sequence several hundred million years ago. PMID- 6298748 TI - Molecular cloning and sequencing of mRNAs coding for minor adult globin polypeptides of Xenopus laevis. AB - Globin mRNA was isolated from immature red blood cells of an adult Xenopus laevis female. mRNA/cDNA hybrids were integrated in the Pst I cleavage site of pBR 322 by G/C tailing, and cloned in Escherichia coli strain HB 101. By restriction site analysis as well as hybridization behaviour we identified two clones coding for minor adult alpha and beta globin chains. Nucleotide sequence analysis and derived amino acid sequences are presented. PMID- 6298750 TI - SV40 deletion mutants lacking the 21-bp repeated sequences are viable, but have noncomplementable deficiencies. AB - We have constructed deletion mutants of simian virus 40 (SV40) lacking the two tandemly repeated copies or all three copies of the 21-bp repeated sequence located in the origin region. The mutants were viable, but had lower infectivities compared to the wild type. The mutant lacking two copies of the 21 bp repeat grew fairly well indicating that the one copy of the 21-bp repeat it contains is adequate. The other mutant lacking all the three copies of the 21-bp repeat was also viable but grew poorly. The viability of this mutant suggests that the upstream 72-bp repeated sequence compensates, though only partially, for the absence of the 21-bp repeat. The growth deficiencies of the deletion mutants could not be overcome by complementation with temperature-sensitive helper mutants providing either the early or the late functions of the virus, suggesting that the deficiencies lie in both early and late gene expression and/or in replication. PMID- 6298749 TI - A cDNA clone containing the entire coding sequence of a mouse H-2Kd histocompatibility antigen. AB - We have isolated a cDNA clone carrying a 1560 bp long insert which contains the entire coding and 3' untranslated regions of an H-2K(d) mouse histocompatibility antigen. Its sequence and overal features are described. They point to the existence of unique properties of DNA sequences associated with the H-2K(d) antigen. PMID- 6298751 TI - Fecal sterols and bacterial beta-glucuronidase activity: a preliminary metabolic epidemiology study of healthy volunteers from Umea, Sweden, and metropolitan New York. AB - The dietary pattern, fecal bile acid and neutral sterol concentrations, and the bacterial beta-glucuronidase activity of 2 population groups with a varied risk for colon cancer development (i.e., a high-risk population in the metropolitan New York area and an intermediate-risk population in Umea, Sweden) were investigated. The average daily intake of dietary protein was the same in the 2 groups, but the fat intake was higher in Umea than in New York. The daily total fiber intake was also higher in Umea, as was the daily total stool output. The concentration of fecal secondary bile acids and beta-glucuronidase activity was lower in Umea than in New York, but the total daily excretion of these constituents was the same in both groups. The data suggest that one of the factors contributing to the lower risk of colon cancer in Umea, despite the high dietary fat intake, is the high intake of dietary whole grain and cereal fiber, which leads to an increase in stool bulk, thus diluting and/or binding promoters. PMID- 6298752 TI - Are there factors preventing cancer development during embryonic life? AB - On the basis of the following literature observations, a hypothesis is advanced that the development of cancer is actively inhibited during embryonic life. Although the processes of cell differentiation and proliferation are--without comparison--most pronounced during embryonic life, cancer is rarely found in the newborn and is seldom a cause of neonatal death or spontaneous abortion. Attempts to induce cancer in early-stage animal embryos by irradiation or by transplacental chemical carcinogenesis have been unsuccessful, even when exposed animals have been observed throughout their life-time. After the period of major organogenesis, however, the embryos become susceptible to carcinogenesis. In humans, the most common embryonic tumors arise in tissues which have an unusually late ongoing development and are still partly immature at or shortly before birth. For many human embryonic tumors the survival rates are higher, and spontaneous regression more frequent, in younger children, i.e. prognosis is age dependent. Thus, although cancer generally appears in tissues capable of proliferation and differentiation, induction of malignancy in the developmentally most active tissues seems to be beset with difficulty. One possible explanation for this paradox could be that cancer is controlled by the regulators influencing development, regulators that are most active during embryonic life. PMID- 6298753 TI - [Information on the parathyroid hormone]. PMID- 6298754 TI - Molecular epidemiology of cytomegalovirus infections in premature twin infants and their mother. AB - Premature identical twins are described who according to molecular fingerprinting of their viral isolates, demonstrate a nonmaternal nursery source for their acquired cytomegalovirus (CMV) infections. The babies were born via cesarean section at 29 weeks gestation. Weekly urine screening of the infants indicated that at birth both were CMV-negative. Twin B developed CMV at 6 weeks of age, while Twin A developed his infection when he was 9 weeks old. Three months following delivery cervical and urine cultures of the infants' mother were negative and she had no detectable CMV antibody. At 6 months postpartum (2 months following both infants' discharge home) a repeat urine culture of their mother was positive for CMV, and here CMV-CF titer had risen to 1:128. DNA fingerprinting by restriction endonuclease digestion analyses of the viruses isolated from the two infants indicate that they were infected with different strains of CMV. In addition the DNA fingerprinting pattern of the mother's isolate is identical to that of Twin A. These cases give further evidence that hospitalized infants may acquire CMV from hospital sources and document by molecular fingerprinting for the first time to our knowledge that these babies may transmit the virus to CMV-seronegative individuals. This study also demonstrates how restriction endonuclease digestion analyses can be used as a powerful tool to study the epidemiology of CMV infections. PMID- 6298755 TI - Fatal adenovirus hepatic necrosis in severe combined immune deficiency. PMID- 6298756 TI - Special series: infectious diseases and the office laboratory. Edited by James K. Todd, M.D. A diagnostic approach to Epstein-Barr virus infections. PMID- 6298757 TI - [Endogenous opiate peptides]. PMID- 6298758 TI - [Various aspects of sodium metabolism in hypertension]. PMID- 6298759 TI - [Procaryotic and viral protein kinases and their biological role]. PMID- 6298760 TI - [The effect of fluorine compounds on respiratory enzymes]. PMID- 6298761 TI - [Application of nucleolytic enzymes in the studies of chromatin structure]. PMID- 6298762 TI - [Significance of elevated serum ACE in pneumology]. AB - Angiotensin-converting enzyme (ACE) levels were measured in the serum of patients with various pulmonary affections (sarcoidosis, tuberculosis, anthracosilicosis, cancer, etc.), with diabetes or renal insufficiency, and in 247 healthy subjects. Mean ACE levels (41.5 U/ml +/- 16) were significantly higher in sarcoidosis than in normal controls (22 U/ml +/- 6), high values (greater than or equal to 34 U/ml) being observed in 62 p. cent of cases. In these patients, the ACE tended to become normal in parallel with clinical and radiological improvement in the disease, and to increase again during relapses. In 75 p. cent of sarcoidosis patients there was concordance between the increase in alveolar lymphocytes and that of serum ACE. Elevated ACE levels were noted in only 3 p. cent of non sarcoidosis pneumopathies, and in 24 and 18 p. cent of diabetes and renal insufficiency cases respectively. In the absence of these two latter affections, measuring ACE levels is of obvious value for the diagnosis and follow-up surveillance of sarcoidosis. PMID- 6298763 TI - Evaluation of Arkansas Rous sarcoma regressor and progressor lines and giant jungle fowl for genetic resistance to the northern fowl mite. AB - Arkansas B1B1R, B2B2R, and B3B3R regressor lines, Arkansas B1B1Pr, B2B2Pr, and B3B3Pr progressor lines, and Giant Jungle Fowl were infested simultaneously at 28 weeks of age with northern fowl mites. The chickens were maintained in similar environments and mite ratings were recorded weekly for 5 months. No strains were completely resistant to mites, but there were significant (P less than or equal to .05) differences among strains in degree of infestation. The regressor lines generally showed significantly (P less than or equal to .05) less infestation than the progressor lines for both female and male chickens. The regressor lines appear to have genetic factors from the Giant Jungle Fowl in the development of the regressor lines that imparted increased resistance to the northern fowl mite. PMID- 6298764 TI - Chloride requirement of young turkeys. AB - Two experiments of 21 days duration were conducted to determine the chloride requirement of the young turkey. Each experiment was repeated once. A corn soybean basal diet with .040% chloride was used. Sodium chloride and sodium bicarbonate were added to the diet to supply .140% sodium and varying levels of chloride. Maximum body weight and feed efficiency were obtained with dietary chloride of .126%. PMID- 6298765 TI - The rachitogenic effects of fractions of rye and certain polysaccharides. AB - When fed to chicks rye is rachitogenic as well as growth depressing. The component or components of rye that cause these effects have not been identified. In an attempt to separate the factors, a water extract of rye was fractionated by precipitation with ethanol or ammonium sulfate. The precipitated fractions were fed to chicks. Although there were different responses to growth and bone ash from the ethanol fractions, they were not statistically significant. In another experiment, guar gum, pectin, or gum arabic was fed to chicks as 2% of the diet. Guar gum was both growth depressing and rachitogenic, pectin was only growth depressing, and gum arabic was without effect. PMID- 6298766 TI - Nutrient deficiencies in broiler chicks: a demonstration project for undergraduate students. AB - A demonstration project is described that proved to be popular with undergraduate students enrolled in a senior level course on animal and poultry nutrition. A total of 100 male, day-old broiler chicks were housed in a Petersime battery brooder and used in a 3-week test. Students were responsible for the management and care of the birds throughout the test. The birds were allotted to 5 groups (4 replicates each of 5 birds per group) and the test involved the omission of calcium, phosphorus, vitamin D3, or sodium chloride from a nutritionally adequate diet for a period of 2 weeks. After that period, one pen of each group was repleted with the control diet. Feed consumption, liveweight, feed conversion efficiency, and tibia bone ash content were measured weekly. Deficiency signs of the nutrients were observed, and it was noted that sodium chloride had the most marked effect on growth. Phosphorus deficiency had the most severe effect on bone development and resulted in a high incidence of rickets. Repletion resulted in a marked response in all parameters measured and was most marked in the group deficient in sodium chloride. PMID- 6298767 TI - Toxicity of a vitamin D steroid to laying hens. AB - Two experiments were conducted with 56-week-old or 104-week-old Leghorn hens to determine if feeding vitamin D steroids in excess of requirement levels caused any marked affects on eggshell quality. In the first experiment caged hens had reduced feed consumption, egg shell quality, and egg production as early as 6 weeks after initially consuming a basal diet supplemented with 6.8 micrograms 1 alpha-hydroxycholecalciferol (1 alpha-OH-D3)/kg. The second experiment confirmed the previous results and showed that extensive weight loss occurred with continued feeding of 10 or 15 micrograms 1 alpha-OH-D3/kg diet. No adverse affects were observed in either experiment when the level of 1 alpha-OH-D3 supplementation was 5.0 micrograms/kg diet or less. No toxic effects were observed when the hormone precursor 25-OH-D3 was supplemented to diets at 6 or 12 micrograms/kg. It is suggested that the pathological effects observed are related to the potent calcium homeostatic properties of 1 alpha-OH-D3 that at elevated levels may cause aberrations in circulating calcium. PMID- 6298768 TI - [Serum angiotensin-converting enzyme and its clinical value in sarcoidosis treatment]. PMID- 6298769 TI - Characterization and partial nucleotide sequence of endogenous type C retrovirus segments in human chromosomal DNA. AB - Twenty-six different murine leukemia virus (MuLV)-related clones have been isolated from a human DNA library and characterized by restriction enzyme mapping and reciprocal nucleic acid hybridization reactions. The sequence of approximately 2,600 nucleotides, spanning more than 4.0 kilobases, of one of the MuLV-related cloned human DNAs was also determined. The deduced amino acid sequence permitted the alignment of this prototype cloned human DNA segment with the p12 gag, p30 gag, p10 gag, and pol regions of Moloney MuLV. A majority of the endogenous type C retrovirus-related segments present in human DNA are approximately 6.0 kilobases in size and appear to contain a deletion of env sequences. PMID- 6298770 TI - Regulation of intramitochondrial cholesterol transfer to side-chain cleavage cytochrome P-450 in rat adrenal gland. AB - Rat adrenal mitochondria accumulated cholesterol during ether stress in vivo when side-chain cleavage was inhibited by aminoglutethimide (control = 14.6 vs. aminoglutethimide = 26.5 micrograms of cholesterol per mg of protein). This accumulation was insensitive to simultaneous administration of cycloheximide (24.2 micrograms/mg), but side chain cleavage in the mitochondria was greatly decreased. Outer and inner mitochondrial membrane fractions were separated by discontinuous Ficoll gradient centrifugation. Quantitation of marker enzymes for inner, outer, and microsomal enzymes indicated that outer membranes contained less than 5% inner membranes. The inner membrane fraction contained less than 7% outer membrane and included 90% of mitochondrial cytochrome P-450. Electron microscopy revealed outer membranes as circular intact ghosts, whereas inner membranes were largely intact and retained vesicular structure typical of intact adrenal cortex mitochondria. Administration of aminoglutethimide effected a 2 fold increase in inner membrane cholesterol (9.4 vs. 20.1 micrograms/mg) but simultaneous administration of cycloheximide completely blocked this increase (10.9 micrograms/mg). We conclude that: (i) in the presence of aminoglutethimide, stress stimulates accumulation of cholesterol in the inner membrane of adrenal mitochondria; and (ii) transfer of cholesterol from outer to inner membranes requires a cycloheximide-sensitive agent. PMID- 6298771 TI - Definition of the simian virus 40 early promoter region and demonstration of a host range bias in the enhancement effect of the simian virus 40 72-base-pair repeat. AB - The simian virus 40 (SV40) origin region includes the viral replication origin and the early and late promoters and consists of a few palindromes, a 17-base pair (bp) A + T-rich sequence, three copies of a G + C-rich 21-bp repeat, and two copies of a 72-bp repeat. We have made sequential deletions in the SV40 origin region and determined the early promoter efficiencies of these truncated DNA segments by connecting them in the correct orientation with the coding regions of selectable marker genes and assaying the expression of the chimeric marker genes in vivo in different host cell lines. A truncated SV40 early promoter segment containing only the TATA box and the major in vivo mRNA initiation sites has essentially no promoter efficiency. We have located the major component of the SV40 early promoter within the 21-bp repeated sequences, which consist of an alternating and mutually overlapping array of two C-rich oligonucleotides having the consensus sequences Y-Y-C-C-G-C-C-C (Y = pyrimidine nucleoside) and G-C-C-C (C)-TA-AT-A(T)-C-T. Between one and two copies of the 21-bp repeat were adequate for gene expression under conditions in which the enhancement effect of the 72-bp repeat was minimal. We also find that the SV40 72-bp repeat exhibits a pronounced host range in its enhancement of gene expression; the enhancement is only 2-fold in the nonpermissive mouse cells but amounts to 10- or 20-fold in the permissive monkey cells or the semipermissive human cells, respectively. PMID- 6298772 TI - Nucleotide sequence of the simian sarcoma virus genome: demonstration that its acquired cellular sequences encode the transforming gene product p28sis. AB - The complete nucleotide sequence of the proviral genome of simian sarcoma virus (SSV), an acute transforming retrovirus of primate origin, has been determined. Like other transforming viruses, SSV contains sequences derived from its helper virus, simian sarcoma-associated virus (SSAV), and a cell-derived (v-sis) insertion sequence. By comparison with the sequence of Moloney murine leukemia virus, it was possible to precisely localize and define sequences contributed by SSAV during the generation of SSV. Comparative sequence analysis of SSV and SSAV showed that SSAV provides regulatory sequences for initiation and termination of transcription of the SSV transforming gene. Moreover, coding sequences for the putative protein product of this gene appear to initiate from the amino terminus of the SSAV env gene. Antibodies to synthetic peptides derived from the carboxy and amino termini of the putative protein predicted by the open reading frame identified within v-sis specifically detect a Mr 28,000 protein, p28sis, in SSV transformed cells. These and other findings confirm the predicted amino acid sequence of this protein and localize it to the coding region of the SSV transforming gene. PMID- 6298773 TI - Degradation and resynthesis of gap junction protein in plasma membranes of regenerating liver after partial hepatectomy or cholestasis. AB - Changes in the total amount of the gap junction protein (M(r) 26,000) after partial hepatectomy or bile duct ligation and recanalization were investigated in rat liver membranes by quantitative immunoblot with rabbit antiserum to the M(r) 26,000 protein. The loss and reappearance of the M(r) 26,000 protein roughly paralleled loss and reappearance of gap junction plaques analyzed previously under similar physiological conditions by freeze-fracture of hepatocyte surfaces. The total amount of the hepatic M(r) 26,000 protein in liver plasma membranes and the total area of the hepatocyte surface occupied by gap junction plaques appeared to be proportional under these conditions. However, at the minimum, 28 35 hr after partial hepatectomy we still find about 15% of the M(r) 26,000 protein, in contrast to <1% of gap junction plaques, determined by morphometric analysis. This discrepancy is probably due to the fact that very small gap junction plaques, single connexons, and free M(r) 26,000 gap junction subunits are missed by the morphometric analysis. At the times of the minimal amount of the M(r) 26,000 protein in hepatic plasma membranes after partial hepatectomy or bile duct ligation we found that crude hepatic lysosomal membranes of these rats contained less M(r) 26,000 protein than lysosomal membranes of nonoperated control animals. Thus, we conclude that the decrease and increase of the total amount of the M(r) 26,000 protein cannot be explained only by dispersal and reuse of gap junction subunits but are largely due to degradation and resynthesis of the M(r) 26,000 protein. No significant change in the amount of the M(r) 21,000 protein that had been isolated with gap junction plaques was observed in liver plasma membranes after partial hepatectomy. This confirms our previous conclusion that the M(r) 26,000 and M(r) 21,000 proteins are independent of each other. PMID- 6298774 TI - Transformation-sensitive protein associated with the cell substratum of chicken embryo fibroblasts. AB - The interaction of cultured cells with their growth substrata has been studied as a function of oncogenic transformation by using chicken embryo fibroblasts infected with the temperature-sensitive mutant of Rous sarcoma virus, LA24, and grown in plastic culture dishes. In comparison to total cell fractions, substratum-associated material (SAM), prepared by EGTA release of transforming cells from culture dishes, is enriched in a 21-kilodalton (kDal) protein. Synthesis and deposition of this protein in SAM are stimulated within hours of transfer of cells to the permissive temperature (35 degrees C), peak around 8 hr, and decline to levels 1.3-fold higher than those of controls at 41 degrees C by 20 hr after the temperature shift. In contrast, incorporation of (3)H-labeled amino acids into newly synthesized fibronectin in SAM is not significantly influenced by the transformation process during this time. Furthermore, although the presence of fibronectin in SAM is influenced by cell density, the 21-kDal protein is increased in SAM of transforming cells at all densities examined. The 21-kDal protein is not present in increased amounts in SAM from normal, uninfected chicken embryo fibroblasts grown at 41 degrees C and 35 degrees C or from cells infected with the wild-type Rous sarcoma virus (Prague A), which are fully transformed. It is not a mannose-containing glycoprotein and does not appear to be phosphorylated. Furthermore, it is not a product of normal cell protein degradation induced by transformation but results from de novo protein synthesis after shift of LA24-infected cells to the permissive temperature. Finally, turnover of the 21-kDal protein is slower at 35 degrees C than at 41 degrees C. This amplifies the effect of increased synthesis and results in a net accumulation in SAM during the early stages of transformation. PMID- 6298775 TI - Identification and characterization of cells deficient in the mannose 6-phosphate receptor: evidence for an alternate pathway for lysosomal enzyme targeting. AB - Newly synthesized lysosomal enzymes acquire phosphomannosyl units, which allow binding of the enzymes to the mannose 6-phosphate receptor and subsequent translocation to lysosomes. In some cell types, this sequence of events is necessary for the delivery of these enzymes to lysosomes. Using a slime mold lysosomal hydrolase as a probe, we have identified three murine cell lines that lack the receptor and one line that contains very low (3%) receptor activity. Each of these lines synthesizes the mannose 6-phosphate recognition marker on its lysosomal enzymes, but, unlike cell lines with high levels of receptor, the cells accumulate oligosaccharides containing phosphomonoesters. The receptor-deficient lines possess high levels of intracellular acid hydrolase activity, which is contained in dense granules characteristic of lysosomes. The data suggest that intracellular mechanisms independent of the mannose 6-phosphate receptor must exist in some cells for the delivery of acid hydrolases to lysosomal organelles. PMID- 6298776 TI - Structural requirement for IS50-mediated gene transposition. AB - Replicative transposition is signaled by the formation of cointegrates in which donor and target replicons are joined by direct repeats of a transposable element. Elements not generating such cointegrates may move by a conservative breaking and joining process. The IS50 elements forming the terminal repeats of Tn5 [which carries the determinant for kanamycin resistance (Kanr)] contain genes and sites necessary for transposition and mediate the movement of any DNA segment they bracket. To determine if IS50 generates cointegrates, the products of transposition from pBR322::Tn5 plasmids to an F factor in recA-Escherichia coli were examined. With monomeric pBR322::Tn5 plasmids, transposition of Kanr (from Tn5) was generally not accompanied by movement of the determinant for ampicillin resistance (Ampr) (from the pBR322 vector). With dimeric pBR322::Tn5 plasmids, by contrast, half of the transpositions of kanr were accompanied by transposition of ampr. Restriction endonuclease analyses indicated that these F-Kanr Ampr chimeras contained inserts of a single copy of the pBR322 vector sequence bracketed by one Tn5 element and one IS50 element or by a pair of Tn5 elements. None of 79 chimeras tested was a true cointegrate. Because IS50 seems to move only a segment of the donor replicon it is proposed that IS50 transposition is conservative. PMID- 6298777 TI - Receptor for transferrin may be a "target" structure for natural killer cells. AB - Human transferrin receptors detected by monoclonal antibody OKT9 appear to be well expressed on cell types known to provide sensitive targets for natural killer (NK) cells. The possibility that transferrin receptors are recognized by NK effector cells has been investigated by three series of experiments: (i) analysis of the correlation between sensitivity to natural killing and the proportion of transferrin receptor-positive cells in different cell lines, (ii) study of the relationship between levels of transferrin receptor expression in cell lines and their capacity to competitively inhibit recognition and killing of the target cell K562 by NK cells, and (iii) comparison of affinity-purified soluble proteolytic fragments of the transferrin receptor and HLA-A, -B molecules for their ability to inhibit the natural killing effect. The data indicate that the transferrin receptor can provide a "target" structure for NK cells. Because transferrin receptors are ubiquitously expressed on normal and malignant proliferating cells, these observations have interesting implications for the possible function of NK cells in vivo. PMID- 6298778 TI - Evolutionary aspects of immunoglobulin heavy chain variable region (VH) gene subgroups. AB - We isolated and determined the sequences of two human germ-line heavy chain variable region (VH) genes and compared them with mouse VH genes. The results show that the human VHI subgroup is evolutionarily related to the mouse VHII subgroup. Evolutionary preservation of homologies in VH genes of the same subgroup includes not only the coding region but also intron size and homology in noncoding regions. This suggests that a VH gene subgroup constitutes a multigene family that undergoes concerted evolution. The homology between genes of the same subgroup in different species is greater than that between genes of different subgroups within a species. One of the VHII genes contains, in complementarity determining region 2 (CDR2), a 13-base-pair previously shown to be in CDR2 of a VHIII gene and in a heavy chain diversity region gene, DH [Wu, T. T. & Kabat, E. A. (1982) Proc. Natl. Acad. Sci. USA 79, 5031-5032], suggesting the insertion of diversity region gene sequences into the VH gene. One of the human VH genes is a pseudogene because of a terminator, which, together with our previous results, shows that the VH gene repertoire contains 40% pseudogenes. In one of the VH genes, direct and inverted repeats at both 5' and 3' ends of the gene suggest a potential transposable element that encompasses the entire VH gene. It is possible that such a structure may facilitate saltatory replication and rapid expansion of VH gene families. PMID- 6298779 TI - Phencyclidine ("angel dust") analogs and sigma opiate benzomorphans cause cerebral arterial spasm. AB - Several psychotomimetic phencyclidine (PCP) analogs--N-ethyl-l phenylcyclohexylamine (PCE), N-[1-(2-thienyl)cyclohexyl]piperidine (TCP), N-[1 (thienyl)cyclohexyl ))pyrrolidine (THP), ketamine, and N,N-dimethyl-l-phenyl cyclohexylamine (PCDEA)--were tested on basilar and middle cerebral arteries of the dog in vitro and found to induce contraction in these blood vessels with a maximal contractile activity (i.e., intrinsic activity) similar to that of PCP. The concentration-effect curves of these compounds were found to be parallel to the curve of PCP (P less than 0.01). The relative potency was PCE greater than TCP greater than PCP greater than THP greater than PCDEA greater than ketamine. A PCP analog with no psychotomimetic activity, 1-piperidinocyclohexanecarbonitrile (PCC), did not induce the blood vessels to contract, nor did the opiate morphine. Three psychotomimetic benzomorphans--pentazocine, cyclazocine, and N allylnorcyclazocine--were found to: (i) also produce contraction; and (ii) have concentration--effect curves parallel to the curve of PCP, but with reduced intrinsic activities (i.e., maximal tensions were lowered) compared to PCP. A kappa opiate, ethylketocyclazocine, relaxed the blood vessels in a dose-dependent manner. This study provides direct evidence for a distinct PCP receptor on cerebral blood vessels and suggests that certain benzomorphans may produce cerebral vasospasm via PCP-receptor interactions. PMID- 6298780 TI - Phosphorylation induces a decrease in the biological activity of the protein inhibitor (GABA-modulin) of gamma-aminobutyric acid binding sites. AB - gamma-Aminobutyric acid (GABA)-modulin is a brain protein of Mr 16,500 that down regulates the high-affinity binding site for GABA which is located in crude synaptic membranes. This protein can be phosphorylated in vitro by the catalytic subunit of cAMP-dependent protein kinase and by a partially purified preparation of calmodulin-sensitive Ca2+-dependent protein kinase. The GABA-modulin sites that are phosphorylated by the two enzymes are different, as revealed by HPLC analysis of tryptic digests. The capacity of GABA-modulin to decrease the number of sites that bind [3H]muscimol was completely abolished by phosphorylation of this protein with the cAMP-dependent protein kinase but not with the Ca2+ dependent enzyme. GABA-modulin present in crude synaptic membranes prepared from rat cortex also was shown to be phosphorylated by endogenous protein kinases activated by cAMP, Ca2+ and calmodulin, and Ca2+ and phosphatidylserine. These results suggest a potentially important role for protein kinase and GABA-modulin in the regulation of the number of GABA recognition sites. PMID- 6298781 TI - The PHI (PHI-27)/corticotropin-releasing factor/enkephalin immunoreactive hypothalamic neuron: possible morphological basis for integrated control of prolactin, corticotropin, and growth hormone secretion. AB - By using the indirect immunofluorescence technique, one and the same neuron in the parvocellular part of the paraventricular nucleus has been shown to stain with antisera against three different peptides: PHI (PHI-27), corticotropin releasing factor (CRF), and enkephalin. This could explain the well-known parallel increase in plasma prolactin, corticotropin, and growth hormone levels- for example, under certain types of stress--as being due to a concomitant release of PHI-like, CRF-like, and enkephalin-like peptides from the same nerve endings in the median eminence. A hypothetical mechanism for the co-ordinated release of these three anterior pituitary hormones is discussed. PMID- 6298782 TI - Abnormal splice in a mutant human beta-globin gene not at the site of a mutation. AB - We have studied the expression of a cloned mutant human beta-globin gene in tissue culture cells. The gene, which was previously isolated from the chromosomal DNA of an individual with a low level of normal beta-globin expression (beta+-thalassemia), contains five mutations inside the large intervening sequence (IVS2), as well as a silent change in codon 2. This beta thalassemia gene (thal) was inserted into a plasmid that is replicated and transcribed in a line of monkey kidney cells in culture. S1 nuclease mapping of the beta-globin RNA transcribed from this gene indicates that some of the beta globin RNA is spliced abnormally by using a cryptic 3' splice sequence normally present in IVS2 but not used in processing the normal beta-globin transcript. The cryptic 3' splice site is not the site of a mutation in the thal gene. Because neither the 5' or 3' splice junction nor the cryptic site is mutated in this gene, it is most likely that the mutation at position 705 of IVS2, the only nonpolymorphic change in the gene, interferes indirectly with normal processing. These results suggest that certain sequences within IVS must be conserved to prevent abnormal splicing and loss of gene function. PMID- 6298783 TI - Insulin regulation of protein phosphorylation in isolated rat liver nuclear envelopes: potential relationship to mRNA metabolism. AB - The direct addition of insulin to highly purified nuclear envelopes prepared from the livers of diabetic rats resulted in a decrease in the incorporation of 32P into trichloroacetic acid-precipitable proteins. Autoradiography of 32P-labeled envelopes, solubilized in sodium dodecyl sulfate and subjected to electrophoresis, revealed that insulin decreased the phosphorylation of all major protein bands. Insulin produced detectable effects at concentrations between 0.1 and 1 pM, maximal effects at 10 pM, and progressively diminished effects at higher concentrations. Two insulin analogs, desdipeptide proinsulin and desoctapeptide insulin, had approximately 10% and 1%, respectively, the activity of native insulin. When nuclear envelopes were first phosphorylated with [gamma 32P]ATP and insulin was then added with an excess of unlabeled ATP, dephosphorylation was enhanced, suggesting that insulin was regulating nuclear envelope phosphatase activity. The direct addition of insulin to isolated rat liver nuclei in the presence of ATP stimulated the release of previously 14C labeled trichloroacetic acid-precipitable mRNA-like material, and the direct addition of insulin to nuclear envelopes stimulated the activity of nucleoside triphosphatase, the enzyme that participates in mRNA nucleocytoplasmic transport. Moreover, the dose-response curves for these functions mirrored insulin's inhibition of nuclear envelope phosphorylation. These data suggest, therefore, a mechanism whereby insulin directly inhibits the phosphorylation of the nuclear envelope, leading in turn to the regulation of mRNA metabolism. PMID- 6298784 TI - Antibodies of predetermined specificity for the NH2 terminus of a cellular protein p53 react with the native molecule: evidence for the presence of different p53s. AB - Two synthetic peptides corresponding to residues 1-20 and 10-20, respectively, of one type of a cellular protein called "p53" have been linked to a carrier protein and injected into rabbits to raise antibodies. The antibodies obtained were capable of reacting with the native protein, as judged by an enzyme-linked immunosorbent assay, protein A-linked staining of immunoblots after NaDodSO4 gel electrophoresis, and immunoprecipitation. The immunoassay titers against the protein were lower for these antibodies than for antisera derived from immunization with purified p53. However, staining with the immunoblot method showed that the antipeptide antibodies against p53 were uniquely specific. The data suggest that at least two different types of p53 molecules occur. The cellular protein previously isolated from human cells transformed by Epstein-Barr virus and from murine tumors induced by methylcholanthrene appears to be larger than the p53 reported in relation to simian virus 40- or adenovirus-transformed cells and to some other tumors. Some interrelationships have not been excluded, but it is clear that the two protein molecules do not behave identically. The reactions of the antipeptide antibodies with the intact protein have implications in regard to protein conformations. The strict specificities of such antibodies allow the generation of distinct sets of reagents useful for quantitation, purification, and cloning. PMID- 6298785 TI - Synthesis of aspartate transcarbamoylase in Escherichia coli: transcriptional regulation of the pyrB-pyrI operon. AB - The first committed reaction in pyrimidine biosynthesis in Escherichia coli and Salmonella typhimurium is catalyzed by the allosteric enzyme aspartate transcarbamoylase (aspartate carbamoyltransferase; carbamoylphosphate:L-aspartate carbamoyltransferase, EC 2.1.3.2), the product of the pyrB-pyrI operon. Regulation of the pyrimidine pathway is achieved in part by changes in the enzyme's catalytic activity as a function of the concentration of substrates and other metabolites as well as by variations in enzyme synthesis in response to changes in cellular levels of pyrimidine nucleotides. Although there is substantial evidence that UTP concentration has a marked influence on expression of the pyrB-pyrI operon, the mechanism of this control is not known. We have cloned the operon and determined the nucleotide sequence of the region preceding the first structural gene (pyrB). These studies show two regions sharing considerable homology with the consensus sequence of E. coli promoters, a segment that can code for a 44-amino-acid leader peptide, and a sequence very similar to that of the attenuator of the trp operon. RNA transcripts from several bacterial strains were studied by S1 nuclease mapping. Under conditions leading to extensive enzyme synthesis there was a large production of transcript whose 5' end correlated with the putative promoter closer to the structural genes. At low levels of operon expression there was little transcript in the extracts and both promoters appeared to serve as initiation sites. The results are interpreted in terms of transcriptional control of the pyrB-pyrI operon according to an attenuation model that differs in novel ways from the mechanisms proposed for the regulation of amino acid biosynthesis. PMID- 6298786 TI - Monensin blocks the maturation of receptors for insulin and somatomedin C: identification of receptor precursors. AB - Cultured human lymphoid (IM-9) cells were labeled with [(35)S]methionine in the presence and absence of monensin, a carboxylic ionophore that inhibits post translational protein maturation. Labeled receptors for insulin and somatomedin C were immunoprecipitated with antibodies specific for each receptor. Monensin inhibits the biosynthesis of mature alpha and beta subunits of both receptors and leads to the accumulation of immunoreactive polypeptides with molecular weights of 180,000. These 180,000 molecular weight polypeptides exist as disulfide-linked dimers and may be biosynthetic precursors of both alpha and beta subunits. In the presence of monensin, small amounts of immunoreactive polypeptides with molecular weights 115,000 and 89,000 also are produced. These may be abnormally processed forms of the alpha and beta subunits lacking residues normally added during terminal glycosylation. In cells treated with monensin, the polypeptides of molecular weights 180,000 and 115,000 can be affinity-labeled with (125)I-labeled insulin. These labeled polypeptides are immunoprecipitated by antibodies specific for insulin receptors but not by antibodies specific for somatomedin-C receptors. This indicates that the putative precursors for insulin and somatomedin-C receptors are distinct polypeptides, although they have similar molecular weights and similar modes of processing. A possible structural relationship between the precursors for these receptors and the type II insulin-like growth factor receptor is discussed. PMID- 6298787 TI - Sharing of biological effect and receptors between guinea pig insulin and platelet-derived growth factor. AB - Insulins from the hystricomorphs (guinea pig, porcupine, coypu, and casiragua) at high concentration stimulate DNA synthesis in human fibroblasts to a greater level than other mammalian insulins or insulin-like growth factors (IGFs). 125I Labeled guinea pig insulin binds to a specific receptor and this binding is competed for by hystricomorph insulins but not by porcine insulin or IGFs. Fetal bovine serum also inhibits the binding of 125I-labeled guinea pig insulin and is more potent than fetal bovine plasma, in concordance with their relative potencies for growth stimulation in human fibroblasts. Of several other known growth factors tested, only platelet-derived growth factor (PDGF) inhibits binding of 125I-labeled guinea pig insulin. Four preparations of PDGF that vary in purity and potency for the stimulation of DNA synthesis in human fibroblasts over a 1,000-fold range compete with binding of 125I-labeled guinea pig insulin in proportion to their biological potencies. The purest preparation of PDGF is able to inhibit binding of 125I-labeled guinea pig insulin by 50% at 15 ng/ml (0.25 nM). Biologically, guinea pig insulin, like PDGF, exhibits a synergistic effect with plasma in initiating DNA synthesis in human fibroblasts; this effect is not observed with other mammalian insulins or IGFs. Thus, hystricomorph insulins appear to be mediating their growth-promoting effect through a different receptor and mechanism than other mammalian insulins or IGFs. Further, hystricomorph insulins may be sharing the mechanism of action for their growth effects with PDGF, perhaps suggesting some relationship between these peptides from very different sources. PMID- 6298788 TI - Growth stimulation of A431 cells by epidermal growth factor: identification of high-affinity receptors for epidermal growth factor by an anti-receptor monoclonal antibody. AB - Epidermal growth factor (EGF) at 3 nM maximally inhibits the proliferation of A431 epidermoid carcinoma cells. We show that at lower concentrations, in the range of 3-100 pM, EGF has a mitogenic effect on A431 cells. In the presence of 100 nM anti-EGF-receptor monoclonal IgG (designated 528), which inhibits A431 cell proliferation and blocks greater than 95% of EGF binding, EGF becomes mitogenic for A431 cells at concentrations up to 3 nM. These results suggest that a minor population of high-affinity EGF receptors may be involved in stimulation of A431 cell proliferation. Saturation binding assays with 125I-labeled EGF indicate that approximately equal to 0.1-0.2% of receptors for EGF are high affinity receptors that bind EGF with an estimated Kd of 7 X 10(-11) M. This affinity is nearly 2 orders of magnitude higher than that of the remaining EGF receptors. Although A431 cell proliferation is maximally inhibited by nonsaturating amounts of EGF (3 nM), maximal inhibition by 528 IgG (approximately equal to 70% of maximal inhibition by EGF) requires saturating concentrations of antibody (approximately equal to 15 nM). Unlike EGF, rapid down-regulation is not observed with 528 IgG. These results indicate different mechanisms of growth inhibition of A431 cells by EGF and 528 IgG. PMID- 6298789 TI - Membrane insertion at the leading edge of motile fibroblasts. AB - We are concerned with the mechanisms involved in the directed migration of eukaryotic cells. Previously we found that, inside cells at the edge of an experimental wound, the Golgi apparatus and the microtubule-organizing center were rapidly repositioned forward of the nucleus in the direction of subsequent cell migration into the wound. This repositioning was proposed to serve the purpose of introducing new membrane mass at the leading edge of the cell, by directing Golgi apparatus-derived vesicles bound for the plasma membrane to that edge. We now provide evidence to support this proposal. Cultured fibroblastic cells at the edge of a wound were infected with a temperature-sensitive mutant (0 45) of vesicular stomatitis virus. It is known that the G-protein, an integral membrane protein of the virus, is synthesized and remains in the rough endoplasmic reticulum at the nonpermissive temperature, but when the infected cells are shifted to the permissive temperature, the G-protein moves through the Golgi apparatus to the plasma membrane. By immunofluorescence microscopy, we here show that the first appearance of the G-protein at the cell surface corresponds to the leading edge of the motile cell. These observations are incorporated into a coherent scheme for the mechanisms involved in cell migration. PMID- 6298790 TI - Isolation and characterization of a temperature-sensitive mutant of avian myeloblastosis virus. AB - A temperature-sensitive (ts) mutant, GA 907/7, was isolated after mutagen treatment of avian myeloblastosis virus. When bone marrow cells or secondary yolk sac macrophages were infected with GA 907/7, the expression of transformation was greatly reduced at 41 degrees C. The results of temperature-shift experiments suggest that in GA 907/7 the putative v-myb gene product is functional only at 35.5 degrees C. Moreover, when ts-induced transformed cells were shifted to 41 degrees C, a partial morphological conversion to macrophage-like cells was obtained, while the majority of the cells underwent senescence and lysis. No leukemia was obtained when GA 907/7 was injected in 1-day-old chickens. Finally, a continuous cell line releasing genetically stable mutant virus was obtained after transformation of secondary yolk sac cells. PMID- 6298791 TI - Identification of adenylate cyclase-stimulating activity and cytochemical glucose 6-phosphate dehydrogenase-stimulating activity in extracts of tumors from patients with humoral hypercalcemia of malignancy. AB - Humoral hypercalcemia of malignancy (HHM) most commonly results from secretion by tumors of an unidentified circulating calcemic factor that appears in clinical studies to stimulate both a parathyroid hormone (PTH)-sensitive proximal tubular adenylate cyclase and a distinct PTH-sensitive renal tubular glucose-6-phosphate dehydrogenase complex. In the present study, 8 M urea extracts of tumors from patients with HHM have been shown to contain both in vitro adenylate cyclase stimulating activity and glucose-6-phosphate dehydrogenase-stimulating activity as detected in a sensitive cytochemical bioassay. Both the adenylate cyclase stimulating activity and cytochemical bioactivity are due to specific binding of a substance in the tumor extracts to renal PTH receptors, as demonstrated by competitive inhibition studies using the bovine PTH fragment analogue [Nle8,18, Tyr34]bPTH-(3-34) amide. Preincubation with an antiserum to PTH results in no loss of activity in the tumor extract, and the activity appears both on gel filtration and ultrafiltration to be far larger than PTH (estimated Mr 70,000). These studies demonstrate that extracts of tumors from patients with HHM contain a substance that binds to the PTH receptors in the nephron responsible for activation of both the PTH-sensitive glucose-6-phosphate dehydrogenase and the PTH-sensitive adenylate cyclase. This substance is chromatographically and immunologically distinct from PTH. Its role in the genesis of HHM requires further study. PMID- 6298792 TI - The neutrophil response to polyvinyl sponge implantation. AB - Neutrophil release and migration in mice were studied over a 24-hr period after the sc implantation of a single polyvinyl sponge. The release of neutrophils from the marrow was evaluated by directly counting the residual neutrophils in the femoral marrow of animals with sponges. Sponge and tissue neutrophil content was determined by extraction and assay of myeloperoxidase (MPO), a marker enzyme for neutrophils. A maximum depletion of 48% of the mature neutrophils in the marrow was observed 5 hr after sponge implantation, in keeping with significant release of neutrophils for migration to the sponge. The released cells were not found in the circulating granulocyte pool, since neutropenia was noted. The accumulation of neutrophils in the sponge increased throughout the 24-hr period, whereas in the tissue adjacent to the sponge maximum accumulation of neutrophils occurred within 7 hr. In fact, neutrophils migrated to at least three sites--the sponge, the skin overlying the sponge, and the skin in which an incision had been made to insert the sponge. The sponge content of neutrophils represented 0.3-33% of the neutrophils migrating to the combined lesion (sponge and skin sites). Therefore, if the neutrophil response to foreign body implantation is to be measured in its entirety, it is necessary to quantify not only the neutrophils within the foreign body but also those in the tissues surrounding it. These studies describe an animal model for neutrophil release and migration to tissues following a standard stimulus. It is proposed that this model may be useful in exploring the factors which influence the release and migration of neutrophils in vivo. PMID- 6298793 TI - Effects of benzo(a)pyrene on Friend viral leukemogenesis in B10SJF1 mice. AB - A single intraperitoneal injection of benzo(a)pyrene (BP) given 1, 2, or 3 days before an ip injection of Friend leukemia virus (FLV) significantly increased the leukemogenic effect of the virus in B10SJF1 mice. These hybrids are the offspring of C57BL/10 females and SJL/J males and are highly resistant to FLV leukemogenesis when the virus is injected alone. PMID- 6298794 TI - Immunosuppression-induced defective lymphocyte proliferation to murine cytomegalovirus is prolonged following the cessation of immunosuppression. AB - Defective murine cytomegalovirus (MCMV) antigen-specific proliferation, induced by treatment of MCMV-infected mice with either antilymphocyte globulin (ALG), prednisolone, or both agents, was eventually restored following the cessation of immunosuppression. At 100 and 278 days following the end of immunosuppressive therapy splenic lymphocytes from infected and subsequently immunosuppressed mice responded significantly in vitro to soluble MCMV antigen after having lost this response immediately upon initiation of immunosuppression. Circulating specific antibodies and mitogen-induced blast transformation were comparable between infected mice and infected mice that also were immunosuppressed. At 278 days following the cessation of immunosuppression splenocytes from infected mice that had been treated with ALG yielded greatly increased background proliferation. Nylon-wool adherence was used to obtain enriched populations of T cells, and B cells and monocytes from MCMV-infected mice. While T cells alone did not respond in vitro to MCMV antigen, recombining B cells and monocytes with the T cells reconstituted in vitro proliferation. Defective lymphocyte proliferation to MCMV for an extended period of time following the end of immunosuppressive therapy indicated a prolonged inability to respond to an active MCMV infection. Identification of the cellular basis for the proliferation defect might lead to the development of effective immunotherapy. PMID- 6298795 TI - Chronic effect of TSH on human thyroid tissue in organ culture. AB - The chronic effect of TSH on thyroidal cAMP concentrations and release of thyroid hormones was investigated using human thyroid tissue in organ culture. Normal human thyroid slices were placed in HAM's F-10 synthetic culture medium in Falcon organ tissue culture dishes, and incubated at 37 degrees in a humidified atmosphere of 5% CO2 in air. Medium was changed everyday and daily T3 or T4 release was determined using concentration of T3 or T4 in the medium. After incubation, slices were transferred to the medium containing 10 mM theophylline and incubated without TSH for an additional 30 min to determine thyroidal cAMP concentrations. Thyroidal cAMP concentrations in slices incubated with 10 mU/ml of TSH increased significantly at 2, 6, and 24 hr and even on the 6th day of incubation. Daily T3 release was significantly increased above control from the 3rd day and daily T4 release from the 4th day to the 11th day of incubation with 10 mU/ml of TSH. Histologically, almost all follicles were structurally maintained even on the 11th day of incubation. These results suggest that both thyroidal cAMP concentrations and release of thyroid hormones are stimulated chronically by TSH. This organ culture system is useful for investigating chronic effects of various materials on human thyroid tissue. PMID- 6298796 TI - Preferential establishment of a slowly dissociable component in plasma membrane compared to intracellular prolactin receptors. PMID- 6298797 TI - Purification and general characterization of FAD synthetase from rat liver. AB - Flavin adenine dinucleotide synthetase (ATP:FMN adenylyltransferase, EC 2.7.7.2) has been enriched more extensively than previously from fresh rat liver. For this, 10% homogenates in sucrose-phosphate buffer were treated with 0.1% Tween-20 prior to high-speed centrifugation to obtain soluble proteins. Those precipitated by 40% saturation with ammonium sulfate were subjected to stepwise addition of calcium phosphate gel to remove pyrophosphatase, and the remaining synthetase was further enriched by passage through a tricalcium phosphate column. An apparent yield of greater than 70% and purification over 70-fold was achieved from the high-speed supernatant fraction. The synthetase activity in solution at 4 degrees was largely lost within a week unless protected by thiols which could partly restore inactivated enzyme. The pH optimum for synthetase activity is near 7.7 when assayed with suitable concentrations of FMN, ATP, and Mg2+. Purified enzyme could be separated into lower (140,000) and higher (325,000) molecular weight components when subjected to molecular sieving on a Sephadex G-200 column. PMID- 6298799 TI - Experimental studies on nutrition, hypertension, and cardiovascular diseases. PMID- 6298798 TI - Dietary influences on lipids and lipoprotein levels in animals and atherosclerosis. PMID- 6298800 TI - The rRNA operon from maize chloroplasts: analysis of in vivo transcription products in relation to its structure. PMID- 6298801 TI - Gene mapping studies and sequence determination on chloroplast transfer RNAs from various photosynthetic organisms. PMID- 6298802 TI - The paromomycin region in the yeast mitochondrial genome. PMID- 6298803 TI - Energy transfer by redox proteins in mitochondria. PMID- 6298804 TI - Evolution of human mitochondrial DNA: a preliminary report. AB - Preliminary mapping of 346 cleavage sites in the mitochondrial genome of 100 human beings gives evidence that the genes for transfer RNA are highly variable, a result that points to the need for testing the accuracy of mitochondrial protein synthesis. In addition, the map comparisons imply that Australia has as much mtDNA diversity as any other area tested in the Old World. Assuming a model of strictly maternal inheritance, it appears possible to follow individual female lineages back hundreds of generations thereby providing human genetics with an important new measure of population heterogeneity. This measure, when compared with those available from protein and morphologic considerations, will help highlight particular groups of people whose mitochondria may increase greatly our understanding of the historical processes leading to the evolution of our own species. PMID- 6298806 TI - The structure and expression of mammalian gene clusters. PMID- 6298805 TI - Interspersed repetitive DNA sequences of the human genome: are they transposons? PMID- 6298807 TI - Properties of a polymorphic DNA segment in the 5' flanking region of the human insulin gene. AB - The 5' flanking region of the human insulin gene contains a section of variable as to length and sequence. This polymorphic region begins 363 bp from the 5' end of the gene and extends upstream for a variable distance. The restriction fragment length heterogeneity is generated by variation in the redundancy of a family of 14-15 bp GC-rich oligonucleotides. The most frequent, as well as the consensus, sequence for this family is ACAGGGGTGTGGGG. The DNA sequence heterogeneity is produced by differences in the arrangement of members of this oligonucleotide family within the tandemly repeating array. On the basis of differences in restriction fragment length, 40 of 74 (56%) unrelated individuals we have examined have been designated heterozygous at this site. The properties of this region make it a useful molecular marker for this region of chromosome 11 as well as loci within 10-20 million base pairs. PMID- 6298808 TI - DNA sequence polymorphism at arbitrary loci. PMID- 6298809 TI - Biochemical genetics of LDL receptor mutations in familial hypercholesterolemia. PMID- 6298810 TI - Cancer susceptibility: family studies of retinoblastoma and Wilms tumor. PMID- 6298811 TI - Toward clinical microcytogenetics: the aniridia and the retinoblastoma stories. AB - In conclusion, the aniridia and the retinoblastoma stories tell us the importance of applying high-resolution cytogenetic techniques and of measuring catalase and esterase D activities when screening and investigating patients and their families. They also point to exceptional situations in carcinogenesis, and without doubt the newly devised genetic techniques, such as recombinant DNA, will allow, when applied to these conditions, a far better understanding of carcinogenesis at the molecular level--which is the important one. PMID- 6298812 TI - Father to son transmission in metaphyseal chondrodysplasia mimicking vitamin D resistant rickets. PMID- 6298813 TI - Syndrome of tibial hypoplasia with polydactyly-syndactyly in two generations. PMID- 6298814 TI - Vitamin D and related research in osteogenesis imperfecta. PMID- 6298815 TI - Pathogenesis of pseudoachondroplasia and diastrophic dysplasia. PMID- 6298816 TI - Histochemical, immunohistochemical, and lectin studies in undecalcified iliac crest growth plate in skeletal dysplasias. PMID- 6298817 TI - Spondyloepiphyseal dysplasia congenita: case report. PMID- 6298818 TI - Against programs: limb development without developmental information. PMID- 6298819 TI - Relationship between the aer, extracellular matrix, and cyclic AMP in limb development. PMID- 6298821 TI - The role of endogenous electrical fields in limb regeneration. PMID- 6298820 TI - Effect of thalidomide-derivatives on limb development in culture. PMID- 6298822 TI - Immunoglobulin anti FSH receptor in the resistant ovary syndrome. PMID- 6298823 TI - Receptors for gonadotropins in human ovaries. PMID- 6298824 TI - Catechol estrogens and pituitary function. PMID- 6298825 TI - Paracrine regulation of the corpus luteum by purines, prostaglandin and peptides. PMID- 6298826 TI - Pineal function in reproductive physiology. PMID- 6298827 TI - Concerning the specificity of the hypothalamic opiate receptor responsible for food intake in the rat. AB - Direct application of small quantities of morphine (a mu-opiate receptor agonist) to the ventromedial hypothalamus (VMH) of rats can induce a stimulated food intake. Because this effect is only partly reduced by the opiate antagonist naloxone, given into the VMH, various other studies were undertaken to examine characteristics of the receptors at this site. The opiate agonist levorphanol but not its stereoisomer dextrorphan effectively increased feeding. Codeine, a weak opiate ligand, was also ineffective as were the kappa-opioid agonist ketocyclazocine and the sigma-opioid agonist phencyclidine. the hyperthermia which accompanies peripheral and central injections of morphine was not observed after hypothalamic application of a quantity of levorphanol sufficient to stimulate feeding. This leads us to propose that the opioid receptors in the VMH are: (1) stereoselective; (2) responsive to mu-, but not kappa- or sigma agonists: and (3) different from the receptors that elicit hyperthermia. PMID- 6298828 TI - Reduced prolactin binding to liver membranes during pheromonal emission in the rat. AB - Between 14 and 27 days of lactation, female rats excrete a pheromone in their feces that is cholic-acid dependent and that strongly attracts young. Previous research has shown that high circulating levels of prolactin are necessary before the pheromone can be emitted. However, during the time of pheromonal emission prolactin in serum conspicuously declines, while in hepatic cytosol the hormone reaches peak levels. We were interested in the question of how the liver can show peak cytosolic concentrations of prolactin at a time of falling blood levels of prolactin. Accordingly, we examined the prolactin binding capacity of liver membrane fractions during selected periods of lactation. We also studied the livers of virgin and pregnant females for comparison. Three membrane fractions were separated: the cell membrane, the nuclear membrane and a fraction consisting of the cell membrane and large non-nuclear organelles. In all three fractions, there was an increase in available and total prolactin binding in the liver when pregnant females were compared with nulliparous females. However, during the time of pheromonal emission, when prolactin in hepatic cytosol was elevated, there was a significant reduction in the prolactin binding capacity of the liver. How such a reduction increases the cytosolic concentration of the hormone and in turn heightens cholic acid output and pheromonal emission remains unsolved. PMID- 6298829 TI - Neuropharmacological profile of R 18 503-induced wet-shake behavior in the rat: noradrenergic and opiate components. AB - R 18 503 (alpha-(p-chlorophenyl)-beta,beta-dimethylimidazole-1-ethanol) 40 mg/kg IP produced a mean of 168 (SEM +/- 4) wet shakes in the 35 min following its administration to 100 g male Wistar rats. No other behavioural abnormalities were seen at this dose. The wet-shakes were not caused by reduced body temperature. However, 20-30 min after R 18 503, body temperature was significantly elevated in comparison with saline treated controls. This supports a role for wet-shakes in thermogenesis in the rat, but suggests that R 18 503-induced wet-shakes are not adapted towards the maintenance temperature homeostasis. R 18 503-induced wet shakes were potently antagonized by 5 narcotic analgesics, but not by aspirin like drugs. A further 18 non-narcotic compounds including putative serotonin antagonists, neuroleptics and compounds thought to act on the alpha-adrenergic system, also antagonized R 18 503-induced wet-shakes. Spectral map analysis showed a close link between ED50-values of the non-narcotic R 18 503 wet-shake antagonists and their ED50's to antagonize noradrenaline-induced lethality; this was further confirmed by a highly significant Spearman Rank correlation between the two tests (rs = 0.77, p less than 0.001). However, the spectral mapping and Spearman correlation analysis showed no relationship between antagonism of R 18 503-induced wet-shakes and antagonism of apomorphine-induced stereotypy, tryptamine-induced bilateral forepaw clonus, mescaline-induced head twitches, and 5 HT-induced contractions of rat caudal artery. It is postulated that R 18 503 induced wet-shakes result from an interaction between the opiate system and the alpha-adrenergic system. PMID- 6298830 TI - Enhanced analgesic response to morphine in adult rats exposed to morphine prenatally. AB - Morphine exposure during development has been shown to produce fetal tolerance to morphine as measured by spontaneous activity only if a particular injection schedule is used. The present study was undertaken to compare the morphine induced analgesic response in adult offspring of rats which had been injected during the last half of gestation on schedules known to produce fetal tolerance (5 mg/kg morphine at 6 hour intervals) versus a schedule known not to produce fetal tolerance (10 mg/kg morphine at 12 hour intervals). At 30 days postnatally the offspring of animals injected on these 2 schedules show no changes in their responsiveness to the analgetic effect of morphine as determined in the hot-plate test. The present study shows that adult offspring of mothers injected with 20 mg/kg/day of morphine in four divided doses on days 12-20 of gestation have an enhanced analgetic response to morphine in the tail-flick test. In contrast, offspring of mothers injected during the same period of gestation with 20 mg/kg/day of morphine in two divided doses respond to the analgetic effect of morphine in the same manner as the offspring of saline-treated mothers. These results show that the schedule for prenatal morphine administration can play a role in the behavioral effects of morphine in adulthood. PMID- 6298832 TI - Limitations to effect of alpha-MSH on permeability of blood-brain barrier to IV 99mTc-pertechnetate. AB - The effects of several variables on the permeability of the blood-brain barrier (BBB) to 99mTc-labeled sodium pertechnetate after IV administration of alpha-MSH were investigated. Doses of alpha-MSH of about 200 micrograms/kg were generally more effective in increasing the brain:blood ratio of radioactivity than the smaller doses that had previously been shown to affect behavior and the EEG. Pulsatile administration of a total of 200 micrograms/kg alpha-MSH over 90 min did not change the permeability of the BBB to the pertechnetate anion. Infusion of the same dose over 90 min significantly increased the brain:blood ratio of radioactivity in one of two experiments: no significant effects were seen with infusion for shorter times, lower concentrations, or with a 4-9 analog (Org 2766). In another experiment, bolus injection of 200 micrograms/kg alpha-MSH resulted in a significantly increased ratio 90 min later as compared with controls. Although the effects of a peptide on the permeability of the BBB to other compounds remains intriguing, limitations appear to exist in experiments with 99mTc-pertechnetate. PMID- 6298831 TI - Opiate effects on social investigatory behavior of male mice. AB - The effects of morphine and naloxone, alone and in combination, on social investigatory behavior and motor activity was examined in CD-1 male mice. Tests were conducted in a Plexiglas apparatus in which a center area was separated from two adjacent stimulus compartments by wire mesh screens. One compartment housed a female conspecific while the other remained empty and served as a control for non specific investigatory responses. A photocell bisected the center compartment and recorded motor activity. Male mice were placed individually into the center area and the time spent investigating each screen was recorded using contact circuits during the 15-min test. In Experiment 1, males (N = 11) received saline, 0.1, 1.0 and 10.0 mg/kg morphine sulfate IP 20 min prior to testing. The high dose significantly decreased investigation of the female compartment while investigation of the uninhibited chamber and motor activity were not significantly affected. In a second experiment (N = 16), 3, 10 and 30 mg/kg naloxone administered 30 min prior to testing had no significant effect on any of the measures recorded. In a third group of subjects (N = 16), 3 mg/kg naloxone reversed the decrease in female investigation time observed with 10 mg/kg morphine, indicating an opiate mechanism for these results. These data provide further evidence that an animal model can be used to study the disruption of socio-sexual behavior produced by opiates. PMID- 6298833 TI - Opposite effects of CRF and ACTH on reserpine-induced hypothermia. AB - The effects of CRF, ACTH 1-24, alpha-MSH, and an ACTH 4-49 analog, at doses of 0, 0.1, 1, and 10 mg/kg, were tested on temperature, ptosis, and sedation in mice pretreated 18 hr previously with reserpine. IP injection of CRF at doses of 1 and 10 mg/kg significantly potentiated the reserpine-induced hypothermia while ACTH 1 24 at the same two doses had the opposite effect of significantly reversing the hypothermia as compared to diluent. The highest dose of alpha-MSH exerted a similar action to that of ACTH 1-24, but none of the doses of the ACTH 4-9 analog changed body temperature. beta-endorphin also failed to cause a reliable effect even though naloxone blocked the action of CRF on body temperature. The results suggest that CRF, like other hypothalamic peptides, can exert extra-pituitary actions after peripheral administration. PMID- 6298834 TI - Hibernation "trigger": opioid-like inhibitory action on brain function of the monkey. AB - A hibernation "trigger" factor derived from the blood of the hibernating woodchuck acts to suppress vital physiological processes in the primate. When infused into the cerebral ventricle of the conscious monkey, the factor induced hypothermia, behavioral depression, bradycardia and aphagia. The opiate antagonists, naloxone and naltrexone, either reverse or retard these behavioral and physiological signs. We hypothesize that the "trigger" molecule is an endogenous opioid-like peptide which may be unique to the hibernator. Moreover, the non-hibernating primate apparently possesses receptor sites in the brain that are capable of responding to this potent molecule. PMID- 6298835 TI - Effects of opiate antagonists and putative mu- and kappa-agonists on milk intake in rat and squirrel monkey. AB - The effects of a number of relatively pure opiate antagonists (naloxone, naltrexone, diprenorphine), and putative mu- (morphine, etorphine) and kappa- (ketocyclazocine, ethylketocyclazocine) receptor agonists on sweetened condensed milk intake were examined over a broad range of doses in non-deprived rats and squirrel monkeys. The antagonists consistently decreased milk intake in both the rat and squirrel monkey. There were, however, species differences: diprenorphine was 30 times more potent than either naloxone or naltrexone in the squirrel monkey, but was of similar potency in the rat. The effects of the opiate agonists were more variable than those of the antagonists. In both species, all agonists decreased milk intake at high doses that also produced behavioral depression. Significant increases in drinking were produced only by low doses of ketocyclazocine and ethylketocyclazocine in the rat. The suppression of milk intake by the antagonists supports a modulatory role of opiate receptors in the control of drinking behavior, however, the effects of the agonists on drinking are less easily interpreted within this conceptual framework. PMID- 6298836 TI - Inhibition by lithium of beta-endorphin-induced psychomotor excitation in cats. AB - beta-Endorphin injected into the cerebral ventricles of unanesthetized cats produced dose-dependent and long-lasting restlessness, locomotion, stereotyped sideways movements of the head, vacant staring, apprehension and flight accompanied with mydriasis and tremor. The most impressive features of the psychomotor excitation were the locomotion and the sideways movements of the head. Intracerebroventricular nalorphine prevented the psychomotor excitation caused by intracerebroventricular beta-endorphin. Lithium chloride and lithium carbonate injected into the cerebral ventricles prevented and reversed the psychomotor excitation evoked by beta-endorphin similarly injected. In cats showing spontaneous locomotor activity, intracerebroventricular lithium chloride also suppressed this activity. It is suggested that beta-endorphin elicited psychomotor excitation by acting on central opiate receptors. However, the effect of lithium cannot be solely ascribed to an action on central opiate receptors and endogenous peptides. Since lithium affected the spontaneous as well as the beta endorphin-induced locomotion, it may be supposed that the cation suppressed the ongoing input activity at central locomotion activity levels. PMID- 6298837 TI - Reversal of morphine-induced catalepsy by naloxone microinjections into brain regions with high opiate receptor binding: a preliminary report. AB - The relationship between opiate binding density and morphine-induced catalepsy was estimated via dose-response analysis of the brain sites in which naloxone microinjections reversed the catalepsy induced by intraperitoneal morphine. One hundred forty-one experimentally naive male Long-Evans rats were implanted with chemical microinjection guide cannulae aimed for various high-to-moderate binding density areas within caudate nucleus, central gray matter, thalamus, hypothalamus, amygdala, and frontal cortex as well as low density sites in pyriform cortex and various fiber tracts. Overall, 48 out of 91 animals microinjected with naloxone in brain sites having high-to-moderate density of opiate binding showed reversal of the cataleptic response. Dose-response effects were found in all 6 high-to-moderate density sites: ranging from 85% reversals at 100 mcg naloxone over all sites to 34% reversals at 0.01 mcg naloxone. There were no reversals out of 38 naloxone microinjection in brain sites having a low density of opiate binding and no reversals out of 18 saline microinjections in either high-to-moderate or low opiate binding density loci. These results suggest a role for limbic and basal ganglia portions of the opiate system in a motor aspect of narcotic action. We speculate that these loci may also play a role in the motor expression of the response to the analgesic and euphoric actions of morphine to supplement actions mediated through periventricular structures. PMID- 6298838 TI - Effects of delta 9-THC and castration on behavior and plasma hormone levels in male mice. AB - Gonadectomy resulted in a rapid increase in plasma luteinizing hormone (LH) and follicle stimulating hormone (FSH) levels, but had no consistent effects on plasma prolactin (PRL) and growth hormone (GH) levels. In castrated males, oral administration of THC (50 mg/kg) significantly increased plasma LH levels within hours following surgery and again from 3 to several weeks post-castration, while THC treatment decreased LH levels between 1 day and 2 weeks postcastration. Administration of THC to 12-hour sham castrates significantly increased plasma LH levels. Plasma FSH, PRL and GH levels were either reduced or unchanged by THC. Administration of THC significantly reduced levels of gonadotropins, PRL and GH in intact males. In additional studies, we examined the influence of THC on the negative feedback response of the anterior pituitary to gonadal steroids. In testosterone propionate (TP)-treated castrated males, concomitant administration of THC increased plasma testosterone (T) and LH at 20 min, while plasma FSH levels were elevated after 60 min. In contrast, in intact TP-treated mice, concomitant THC exposure reduced plasma T levels except at 60 min, when plasma LH levels were significantly increased. Testosterone replacement failed to restore copulatory behavior in castrated mice given a single dose (50 mg/kg) of THC. In fact, acute THC administration to these TP-treated castrates resulted in marked sedation, which was not observed in intact mice given the same dose of THC in an earlier study. The present findings indicate that the effects of acute THC treatment on pituitary gonadotropin release is dependent upon the time after castration. Furthermore, THC administration can suppress copulatory behavior even in animals whose peripheral T levels have been maintained. Effects of THC on plasma androgen levels in animals injected with TP suggest that THC can alter the metabolism or target tissue response to gonadal steroids. PMID- 6298840 TI - [Synthesis and properties of digitoxigenin-3 beta-O-alpha-L-arabinofuranoside]. AB - The synthesis of a cardenolide glycoside with a furanoide sugar component, digitoxigenin-3 beta-O-alpha-L-arabinofuranoside (4), is described for the first time. 4 was prepared by the reaction of digitoxigenin with 2,3,5-tri-O-benzoyl alpha-L-arabinofuranosylchloride and Fetizon-reagent in benzene/dioxan followed by debenzoylation with ammonia in dry methanol. Compound 4 is cleaved by alpha-L arabinofuranosidase (Aspergillus niger K1) into digitoxigenin and L-arabinose. Hydrolytic stability against methanolic HCl (0.1 mol/l) is relatively high. 4 X 10(-8) mol/l 4 gives a 50% inhibition of the Na,K-ATPase (pig heart muscle) and is 2.5 times more active at this receptor than the aglycon digitoxigenin. PMID- 6298839 TI - Cold stress in the rat induces parallel changes in plasma and pituitary levels of endorphin and ACTH. AB - Endorphin and ACTH-like materials levels in rat plasma and pituitary were measured by radioimmunoassay under baseline and cold stress conditions. Cold stress significantly increased plasma beta-endorphin and ACTH immunoreactivity. A rise in these two peptides was also found in the neurointermediate lobe of the pituitary, while in the anterior lobe their levels were unaffected. These findings suggest that the rise of beta-endorphin and ACTH content in the neurointermediate lobe occurs as a compensatory biosynthetic mechanism for the peptides released from the adenohypophysis. PMID- 6298841 TI - Effects of delta 9-tetrahydrocannabinol, cannabinol and cannabidiol on the immune system in mice. I. In vivo investigation of the primary and secondary immune response. AB - The effects of the cannabinoids delta 9-tetrahydrocannabinol (THC), cannabinol and cannabidiol on the primary humoral immune response, the secondary humoral immune response and the memory aspect of humoral immunity in response to sheep red blood cell (SRBC) immunization was investigated. Mice treated with THC (10 and 15 mg/kg) during the primary immunization period exhibited a suppression of the primary humoral immune response. Mice treated with THC during the secondary immunization period showed no measurable suppression of the secondary humoral immune response to the immunizing antigen. The memory aspect of humoral immunity was assessed when treatment with cannabinoids was carried out during the primary immunization period and the ability of mice to undergo a secondary immune response was evaluated; suppression of the secondary humoral immune response was evident with THC treatment (10 and 15 mg/kg). Cannabinol and cannabidiol (10 and 25 mg/kg) treated mice showed no impairment in the ability to undergo primary or secondary immune responses with any treatment protocol. In vivo investigations of the effects of cannabinoids on the thymus were also carried out. Thymus weight and thymus cell number were depressed in mice undergoing a primary humoral immune response when treated with THC (10 and 15 mg/kg) during this period. THC treatment, however, did not alter these parameters in mice not challenged with antigen. In both challenged and unchallenged animals, cannabinol and cannabidiol did not measurably alter the thymus. PMID- 6298842 TI - Effects of delta 9-tetrahydrocannabinol, cannabinol and cannabidiol on the immune system in mice. II. In vitro investigation using cultured mouse splenocytes. AB - The effects of the cannabinoids, delta 9-tetrahydrocannabinol (THC), cannabinol (CBN) and cannabidiol (CBD), on the primary-like immune response were investigated in primary cultures of mouse splenocytes. Splenocyte cultures were stimulated with sheep erythrocytes in vitro and incubated with cannabinoids for the first 24 h after antigenic stimulation (prior to initiation of DNA synthesis), from 24 h to 6 days after antigenic stimulation, and for the entire 5 day period. THC (1 and 5 microM) and CBD (5 microM) depressed the primary-like immune response of stimulated mouse splenocytes when incubated for the first 24 h after antigenic stimulation and the entire 6-day culture period. CBN did not show any measurable suppression of the primary-like immune response. Treatment of splenocyte cultures with cannabinoids after the first 24 h after antigenic stimulation showed no impairment of the in vitro primary-like immune response. PMID- 6298843 TI - Hormonal responses accompanying fear and agitation in the squirrel monkey. AB - The adrenocortical and gonadal responses of 14 male monkeys were evaluated during four experimental conditions in order to evaluate the influence of social interactions on endocrine responsiveness. Plasma hormone levels were determined during the establishment of social relations, after 60-min exposures to a novel environment, after 60-min exposures to a snake, and 60 min after ACTH administration. Both adrenal and gonadal secretion changed significantly during the first day after social relations were established, although only dominant males showed increases in testosterone, whereas cortisol levels rose in all subjects. Increases in cortisol, but not testosterone, were also observed following exposure to novelty or a snake. The presence of a social partner reduced signs of behavioral disturbance during these test conditions, although the adrenal responses were equivalent or greater than when tested alone. This finding qualifies earlier research which indicated that social support was beneficial for reducing stress when squirrel monkeys were tested in larger groups in their home environment. PMID- 6298845 TI - Restricting eye movements in the anesthetized rat: a comparison of four techniques. AB - Four techniques for the immobilization of rodent eyes were compared. Eye movements were plotted over a three hour period by reflecting a laser beam off a small mirror affixed to the cornea. Surgical techniques for producing local paralysis of the extra-ocular eye muscles were inconsistently successful. Physical stabilization techniques reliably reduced eye movements to an extent compatible with certain neurophysiological tests. None of the techniques was as efficacious as the use of neuromuscular blocking agents combined with physical restraint. PMID- 6298844 TI - Memory consolidation: further evidence for the four-phase model from the time courses of diethyldithiocarbamate and ethacrynic acid amnesias. PMID- 6298846 TI - Collagenase in appropriate concentrations and nonspecific proteases do not interfere with the adrenergic responsiveness of adipocytes. PMID- 6298847 TI - Detection of pericine, a new CNS-active indole alkaloid from Picralima nitida cell suspension culture by opiate receptor binding studies. PMID- 6298848 TI - Follow-up on lacrymal gland adenoid cystic carcinoma. PMID- 6298849 TI - Naloxone, tardive dyskinesia, and endogenous beta-endorphin. AB - The subjects were 13 psychiatric inpatients with tardive dyskinesia. Each subject participated in two sessions. Either naloxone (10 mg) or placebo was administered intravenously during each session. In a subset of subjects (n = 7), blood samples for beta-endorphin were drawn before and at 30 and 60 minutes after the injection. The Abnormal Involuntary Movement Scale was administered before and at 10, 20, 40, 60, 120, and 360 minutes after the injection. Double-blind procedures were maintained throughout the experiment. Neither naloxone nor placebo had any appreciable effect on the involuntary movements. Naloxone elicited a significant increase in the plasma beta-endorphin. PMID- 6298850 TI - Putative PEA receptors? PMID- 6298851 TI - Temporal distribution of myocardial infarction pain. PMID- 6298852 TI - Adrenal cortex of fetal lamb: changes after hypophysectomy and effects of Synacthen on cytoarchitecture and secretory activity. AB - The effects of hypophysectomy on the cell population of the adrenal cortex has been examined with light and electron microscopy. The sensitivity of the adrenal cortical cells to exogenous ACTH in both normal and hypophysectomized fetuses has been investigated by comparing plasma cortisol concentrations with structural changes in the adrenal cortex. Hypophysectomy was carried out at about 100 d and the subsequent analysis was made at about 135 d on fetuses catheterized at least 6 d earlier. The zona fasciculata of intact fetuses contained about 25% mature and 75% immature cells at 130-136 d. After hypophysectomy this zone contained a uniform population of immature cells and no mature cells; there was a significant reduction in both adrenal weight and cortical thickness. The zona glomerulosa was unaffected by hypophysectomy. The infusion of Synacthen 0 . 25 mg/d I.V. for 48 h did not affect the zona glomerulosa but resulted in a 2-fold increase in the thickness of the zona fasciculata in hypophysectomized and intact fetuses. This cortical growth was due to both hypertrophy and hyperplasia. All the cells in the zona fasciculata were mature following Synacthen treatment in both groups. The most striking difference between the secretory response of the adrenal cortex to Synacthen in the two groups was found within 3-6 h of the onset of infusion. Within this period there was a significant increment in the plasma cortisol concentration in the intact fetuses while no response was observed in the hypophysectomized animals. After 12 h infusion by contrast, a steadily rising response was observed in both groups. There was a significant correlation between the final plasma cortisol concentration and both the adrenal weight and cortical thickness after 48 h treatment. The increased sensitivity of the adrenal cortex to exogenous ACTH as term approaches in the sheep is considered to be dependent upon the increasing proportion of mature zona fasciculata cells within the adrenal cortex. PMID- 6298853 TI - Radiation effects on bovine taste bud membranes. PMID- 6298854 TI - [Comparative radioprotective activity of adenylates in short-term and prolonged gamma-irradiation]. PMID- 6298855 TI - [Mechanism of chromatin degradation in thymocytes of irradiated rats. 5. Effect of irradiation and cycloheximide on the sensitivity of chromatin to exogenous nucleases]. PMID- 6298856 TI - Wilms tumor in horseshoe kidneys: radiologic diagnosis. AB - Two cases of nephroblastoma occurring in a horseshoe kidney are reported, and 32 cases from the literature are reviewed. The radiologic signs of horseshoe kidney may be difficult to evaluate with excretory urography when the mass is large. Rotational abnormalities of the opposite kidney that is not involved by tumor should suggest the possibility of an associated horseshoe kidney. Real-time ultrasonography and computed tomography are helpful in identifying the isthmus of the horseshoe kidney. Aortography confirms the presence of the horseshoe kidney and demonstrates the arterial supply to the isthmus and the tumor. Radionuclide scans demonstrate the isthmus when the tumor arises from an upper pole, but may not be diagnostic if the tumor arises from the isthmus. PMID- 6298857 TI - Work in progress: dynamic sequential computed tomography during arterial portography in the detection of hepatic neoplasms. AB - Dynamic sequential computed tomography with table incrementation during arterial portography (DSCTI-AP) of the entire liver was performed in an attempt to improve the detection of hepatic neoplasms. In 13 of 17 patients, DSCTI-AP detected more hepatic lesions than were detected by other imaging methods, including radionuclide liver scans, ultrasound, computed tomography, and hepatic angiography. In only one case, an extremely hypervascular hepatocellular carcinoma, hepatic angiography was superior to DSCTI-AP. DSCTI-AP is a simple technique that may be carried out as a part of hepatic angiography. We believe that DSCTI-AP is a sensitive and useful method for the accurate detection of hepatic neoplasms. PMID- 6298858 TI - Ultrasound in the evaluation of solid breast masses. AB - Ultrasound image characteristics and radiographic features of 31 benign and 41 malignant breast masses were cross-tabulated and analyzed to determine the ultrasound image characteristics most useful for diagnosis and the frequency with which some imaging features occurred. The most common malignant mass (the ductal carcinoma) exhibited a jagged wall (88%), homogeneous internal echoes (12%), nonhomogeneous internal echoes (70%), internal echoes not discernible due to attenuation effects (18%), and attenuation shadowing (97%). In contrast, the fibroadenoma (the most common benign mass in this study), exhibited smooth walls (94%), homogeneous internal echoes (89%), and no demonstrable posterior shadowing (67%). PMID- 6298859 TI - Differentiation of focal intrahepatic lesions with 99mTc-red blood cell imaging. AB - The appearance of focal hepatic lesions on 99mTc-sulfur colloid images is nonspecific. As it is important to distinguish hemangiomas from other lesions prior to biopsy, a prospective study was performed using 99mTc-labeled red blood cells. Dynamic perfusion and delayed blood-pool images (1-2 hours) were obtained and lesion activity categorized as increased, equal, or decreased compared with the liver. Of 21 patients studied, 9 (43%) had one or more hepatic hemangiomas, and 8 of these 9 patients (89%) demonstrated increased blood-pool activity. The 12 nonhemangiomatous lesions consisted of 7 metastatic tumors, 2 hepatomas, 1 cirrhotic nodule, and 2 hepatic cysts. None of these 12 patients had increased activity on delayed blood-pool images. Early dynamic images of hepatic hemangiomas demonstrated variable activity (vascularity) and were not useful in differentiating hemangiomas from other lesions. Sensitivity was 89% and specificity 100%. Although liver enzymes are usually normal with hepatic hemangiomas, they may also be normal in metastatic disease. The authors recommend that delayed blood-pool imaging be performed prior to biopsy, particularly in patients without a known primary tumor or those with normal liver enzyme levels. PMID- 6298860 TI - Work in progress: transcatheter management of primary carcinoma of the liver. AB - Twenty-four patients with hepatocellular carcinoma or cholangiocarcinoma were treated with hepatic artery infusion (HAI) of chemotherapeutic agents (14 patients) or hepatic artery embolization (HAE) (11 patients). One patient had a combination of the two treatments. Ten of the 14 (71%) patients who underwent HAI demonstrated partial remission. The median survival time was 12.3 months. Six of the 9 patients (67%) who underwent HAE and for whom follow-up was available responded to treatment. The median survival time for those patients was 17.4 months. PMID- 6298861 TI - Herpes infection: nurse sued doctor. Case in point: Livingston v. Gribetz, M.D. (549 F. Supp. 239 - N.Y.). PMID- 6298862 TI - Chemical transmission in the brain. The role of amines, amino acids and peptides. Proceedings of the 12th International Summer School of Brain Research, Amsterdam, The Netherlands, August 31 to September 4, 1981. PMID- 6298863 TI - beta-Endorphin in brain. PMID- 6298864 TI - Plasticity in synaptic transmission and changes of membrane-bound protein phosphorylation. PMID- 6298865 TI - Dopamine neurotransmission and brain function. PMID- 6298866 TI - Pro-opio(melano)cortin and brain homeostasis. PMID- 6298867 TI - Aspects of protein phosphorylation in the nervous system with particular reference to synaptic transmission. PMID- 6298868 TI - Cyclic GMP-dependent protein phosphorylation in mammalian cerebellum. PMID- 6298869 TI - The regulation of enzyme activity by reversible phosphorylation. PMID- 6298870 TI - Substrates of nuclear protein kinases in rat C6 glial cell cultures. PMID- 6298871 TI - Phosphorylation in relation to the modulation of brain protein synthesis by ACTH like neuropeptides. PMID- 6298872 TI - Role of calmodulin in brain function. PMID- 6298873 TI - Ca2+-calmodulin tubulin kinase system and its role in mediating the Ca2+ signal in brain. PMID- 6298874 TI - Membrane phospholipid turnover, receptor function and protein phosphorylation. PMID- 6298876 TI - Modulation of adenylate cyclase by protein phosphorylation: effects of ACTH. PMID- 6298875 TI - Protein phosphorylation in the regulation and adaptation of receptor function. PMID- 6298877 TI - ACTH and synaptic membrane phosphorylation in rat brain. PMID- 6298878 TI - The modulation of protein phosphorylation and receptor binding in synaptic membranes by changes in lipid fluidity: implications for ageing. PMID- 6298879 TI - Proposed role of phosphoproteins in visual function. PMID- 6298880 TI - The link between analgesia and cardiovascular function: roles for GABA and endogenous opioids. PMID- 6298881 TI - Endogenous opioid peptides: review of physiological, pharmacological and clinical aspects. PMID- 6298882 TI - Aspects on the physiology and pharmacology of the bladder and urethra. PMID- 6298883 TI - Adult neurons in cell culture: electrophysiological characterization and use in neurobiological research. PMID- 6298884 TI - CNS putative L-prolyl-L-leucyl-glycinamide (PLG) receptors, brain and lymphocyte dopamine receptors. AB - 1. In view of previously demonstrated modulatory effects of PLG on the sensitivity of central dopamine receptors, we developed a radioligand binding assay to identify specific binding sites of PLG in rat and normal human brain. 2. 3H-PLG binds specifically to rat striatum exhibiting high affinity (KD = 4.69 +/- 0.50 nM) saturability (Bmax = 9.20 +/- 0.30 fmoles/mg protein) and reversibility; the highest density of specific PLG binding sites occurring in the striatum, followed by the hypothalamus and cerebral cortex. 3. Saturable, high-affinity binding sites of PLG were identified in human striatum. The substantia nigra was enriched with the highest level of specific PLG binding sites. 4. Dopamine receptors were identified in human lymphocytes. 5. The results are compatible with the hypothesis that differential modulation of CNS dopamine receptors by PLG is functionally associated with interacting with specific PLG binding sites in the rat and human brain, and pose implications for Parkinson's disease and tardive dyskinesia. PMID- 6298885 TI - 6-Hydroxydopamine-induced blockade of hypothalamic obesity: critical role of brain dopamine-norepinephrine interaction. AB - 1. Adult female rats received 3 weekly intracisternal injections of 6 hydroxydopamine or ascorbic acid vehicle following systemic pretreatment with desipramine or pargyline to deplete brain dopamine alone or in combination with norepinephrine. 2. Three weeks after the last injection, all rats received bilateral radiofrequency lesions of the medial hypothalamus to induce overeating and obesity. 3. Compared to normal controls, all lesioned groups over ate and became equally obese except the group receiving desipramine + 6-hydroxydopamine, which failed to show either response. 4. Terminal assays of striatal and neocortical monoamines revealed that hypothalamic obesity was blocked only when dopamine was selectively depleted, but not when equivalent dopamine depletion occurred with concurrent norepinephrine depletion. 5. The ability of central 6 hydroxydopamine to block hypothalamic obesity seems due to an imbalance created in the neural interactions of brain dopamine and norepinephrine systems. PMID- 6298886 TI - Interaction of beta-carbolines with the benzodiazepine receptor. Structure activity relationships of amide derivatives of beta-carboline and tetrahydro-beta carboline. AB - 1. The inhibition of specific 3H-flunitrazepam binding to rat cortical membrane preparations (benzodiazepine receptors) by a series of amide derivatives of beta carboline and tetrahydro-beta-carboline related to the ethyl ester of beta carboline 3-carboxylate (beta-CCE) was measured. 2. beta-Carboline amides which are unsaturated in the C ring of the beta-carboline nucleus were the most potent inhibitors of benzodiazepine receptor binding. 3. Increasing the length of the hydrocarbon moiety in the aliphatic amide side chain beyond two carbon atoms decreased potency. PMID- 6298887 TI - beta-Adrenergic, CRF-ergic and dopaminergic mechanisms controlling alpha-MSH secretion in rat pars intermedia cells in primary culture. AB - 1. A pure population of pars intermedia cells in primary culture was used to study changes in alpha-MSH secretion and cyclic AMP accumulation. 2. Beta adrenergic agonists and CRF (corticotropin-releasing factor) stimulate alpha-MSH secretion and cyclic AMP accumulation. 3. Dopaminergic agonists inhibit basal as well as (-)isoproterenol- and CRF-induced alpha-MSH secretion and cyclic AMP accumulation. 4. Beta-adrenergic, dopaminergic and CRF receptors regulate pars intermedia cell activity probably through the adenylate cyclase system. PMID- 6298888 TI - Alpha 1-adrenergic stimulation of ACTH secretion in vivo in the rat. AB - The alpha-adrenergic agonist phenylephrine (100 micrograms, i.v.) causes a rapid 5- to 10-fold elevation of plasma ACTH levels 5 min after its administration in adult ovariectomized rats, the concentration of the hormone remaining elevated up to at least 2h. Epinephrine (10 micrograms) causes also a rapid but shorter-lived stimulation of ACTH secretion. While having no effect alone under basal conditions (conscious freely-moving animals), the highly specific alpha 1 adrenergic antagonist prazosin (0.25 micrograms) almost completely reverses the stimulatory effect of phenylephrine. Pretreatment with dexamethasone inhibits basal plasma ACTH levels by 70% and almost completely prevents the stimulatory effect of phenylephrine on this parameter. Plasma levels of alpha-MSH, on the other hand, are only stimulated 1-fold above control 5 min after the administration of phenylephrine and are insensitive to corticosteroid treatment. Based on the specificity of action of prazosin on postsynaptic alpha 1-adrenergic receptors and of dexamethasone on the anterior lobe of the pituitary gland, the present data indicate that phenylephrine is a potent stimulator of ACTH secretion by a direct action on an alpha 1-adrenergic receptor in corticotrophs of the adenohypophysis. They also support the suggestion that epinephrine and/or norepinephrine could be involved as physiological corticotropin-releasing factor(s). PMID- 6298889 TI - Effect of chronic lithium treatment on twenty four hour variation in plasma and red blood cell lithium and sodium concentrations, drinking behavior, body weight, kidney weight, and corticosterone levels. AB - This study tested the hypothesis that lithium alters the phase relationships of various 24-hour rhythms. Six point 24-hour patterns were measured throughout a 12 hour light/12 hour dark cycle from separate groups of individually housed adult male wistar rats maintained for six weeks on ad lib water and either 1) Normal lab chow, 2) Lab chow supplemented with 50mM/KG of lithium chloride, or 3) Lab chow supplemented with 50mM/KG of sodium chloride. Plasma lithium levels were 0.7 1.0 mEq/1. The sodium diet had no effect on any of the variables measured relative to normal controls. Plasma but not red blood cell lithium levels demonstrated a 24-hour rhythm with higher levels during darkness. Serum and red blood cell sodium levels did not differ among the diets and showed no 24-hour variation. Lithium treated rats weighed significantly less than controls and had significantly heavier kidneys per 100 grams of body weight than controls. Resting plasma corticosterone levels demonstrated the expected 24-hour pattern in all groups, however, the lithium group evidenced higher levels during the middle of the dark hours than either control group. Water consumption also demonstrated a 24-hour rhythm. Lithium animals consumed far greater quantities of water than controls and this behaviour became increasingly exaggerated over the duration of the study. The data are interpreted as suggesting that lithium broadens the peak of twenty four hour rhythms such that normal elevation in these measures appear earlier and last longer. PMID- 6298890 TI - U-50,488, a selective kappa opioid agonist: comparison to other reputed kappa agonists. AB - 1. U-50,488 is a structurally novel, non-mu opioid. In the present experiments it was compared to the reputed kappa opioid agonists, ketazocine, ethylketocyclazocine and bremazocine as regards analgesic cross tolerance to morphine and U-50,488, antagonism of analgesia by naloxone and MR-2266 (in vivo pA2 determination), and narcotic antagonist properties (antagonism of morphine analgesia and precipitation of abstinence in morphine-dependent mice). 2. The analgesic mechanism of bremazocine was similar to that of U-50,488 but the former compound had, in addition, considerable mu-antagonist activity. The analgesic mechanisms of the ketazocines were less selective; both shared both mu and kappa agonist properties. U-50, 488, however, had no such mu agonist or antagonist effects and thus is a more selective kappa agonist. 3. This compound and its congeners may prove useful in the elucidation of the functions of kappa receptors in the central nervous system. PMID- 6298891 TI - Gabaergic regulation of nigrostriatal neurons: coupling of benzodiazepine and GABA receptors. AB - 1. The GABA agonist muscimol and the benzodiazepine diazepam both decrease dopamine (DA) release in the rat nigrostriatal pathway. 2. These actions are reversed by the GABA antagonist picrotoxin and in the case of diazepam, by the benzodiazepine antagonist RO151788. 3. Our data support an inhibitory GABAergic regulation of nigrostriatal DA neurons and indicate the presence of a GABA/benzodiazepine receptor complex on these neurons. PMID- 6298892 TI - Some aspects of basic psychopharmacology: the trace amines. AB - 1. Some recent research on the behavioural effects of phenylethylamine and some recent data implicating the trace amines in schizophrenia, agoraphobia and aggression are briefly outlined. 2. Phenylethylamine produces in mice a distinctive hyperactivity syndrome consisting of two phases; it appears to act via dopamine and 5-hydroxytryptamine on different components of these stereotypies. 3. Urinary unconjugated tryptamine, and meta- and para-tyramine appear to be excreted in reduced amounts in schizophrenia and bipolar depression. 4. The blood levels of the trace acids phenylacetic and meta- and para hydroxyphenylacetic are reduced in schizophrenia. 5. Blood levels of conjugated phenylacetic and unconjugated para-hydroxyphenylacetic acid are reduced in violent as opposed to non-violent offenders. 6. The neuromodulatory role of the trace amines and their possible involvement in components of behaviour and certain mental disorders are discussed. PMID- 6298893 TI - Enkephalin metabolism in brain and its inhibition. AB - Various cerebral peptidases are able to hydrolyse the enkephalins into inactive fragments. Among these enzymes enkephalin-dipeptidylcarboxypeptidase ("enkephalinase") inhibited by thiorphan and a bestatin-sensitive aminopeptidase activity seem to play a key-role as "inactivating neuropeptidases". Their inhibition, in vitro as well as in vivo, leads to a protection of endogenous enkephalins and to antinociceptive effects. PMID- 6298894 TI - [Structural analysis of mutant genes in mammalian somatic cells]. PMID- 6298895 TI - [Cell functions and calmodulins in protozoa. (I). Activation by calmodulin of various enzymes]. PMID- 6298896 TI - [Calmodulin and physiological functions of plants]. PMID- 6298898 TI - [Ca-ATPase from red blood cell]. PMID- 6298897 TI - [Regulation of Ca2+-dependent ATPase of cardiac sarcoplasmic reticulum by cAMP- and calmodulin-dependent phosphorylation of phospholamban]. PMID- 6298899 TI - [Ca-ATPase from synaptic plasma membrane]. PMID- 6298900 TI - [Oncogenicity of adenoviruses]. PMID- 6298901 TI - [Recombinant DNA technology and its use in the study of adenovirus]. PMID- 6298902 TI - Vascular reactivity and high dietary eicosapentaenoic acid. AB - Epidemiologic studies suggest that high dietary intake of eicosapentaenoic acid (EPA), a precursor of the trienoic prostaglandins, is associated with a low incidence and reduced extent of myocardial infarction. Vascular reactivity of isolated aortic strips from rats maintained for 3 weeks on a control diet or on a diet supplemented with menhaden fish oil (17% EPA) was examined with norepinephrine, sodium arachidonate, KC1, PGF2 alpha and nitroprusside. Aortic strips from rats fed the fish oil diet were significantly less responsive to the contractile effects of norepinephrine and arachidonate compared to those from control diet rats. Treatment of aortic strips with indomethacin decreased responsiveness to norepinephrine. The magnitude of the decrease was greater in control rats resulting in a similar vascular response between the 2 groups after blockade. Contractions to arachidonate were abolished by indomethacin. There were no differences in vascular responses to KC1, PGF2 alpha and nitroprusside in aortic strips from control diet rats and those from the fish oil diet rats. Aortic strips from the fish oil diet rats contained more EPA than those from the control diet rats. Thus, the contractile effect of norepinephrine in isolated rat aortic strips is normally augmented by intrinsic prostaglandins, and this augmentation is diminished by dietary intake of EPA. PMID- 6298903 TI - "Menstrual induction" with Sulproston. AB - The PGE2-analogue Sulproston (16-phenoxy-omega-17,18,19,20-tetranor-PGE2 mythylsulfonylamide) was administered to 200 medically and gynecologically normal women who were 17 +/- 0.4 days beyond their expected menstrual period and who had a positive pregnancy test. The intramuscular impact dose (500 micrograms repeated after 4 hours) caused an immediate tonic uterine contraction which compromised the estradiol 17 beta, progesterone and chorionic gonadotropin production within the fetoplacental unit, and thereby allowed the evolution of cyclic uterine activity, cervical dilatation and tissue expulsion. Pregnancy termination was complete in 92% of women, 5.5% required surgical curettage and 2.5% were given a second Sulproston treatment 2-3 weeks after the first to remove retained tissue from the uterus. The medical induction of menstruation was preferred by 83% of the women who had previously experienced surgical termination of pregnancy. Normal menstruation resumed in all women after 36 +/- 0.9 days. The majority of 42 women questioned found Sulproston a satisfactory, safe, simple and effective drug regimen for "menstrual induction". PMID- 6298905 TI - Approved HV1 and HV2 systems. PMID- 6298904 TI - The human placental anti-aggregating factor is neither prostacyclin, nor a prostacyclin metabolite. AB - Using PGH2 as substrate, we have previously demonstrated that human placenta synthesizes mainly PGE2, TxB2 and PGD2(1,2). Other reports have shown that placental tissue generates a substance which inhibits ADP-induced platelet aggregation and which was supposed to be PGI2 (3). The present study indicates that the stability of that substance is different from the stability of prostacyclin (released by umbilical artery pieces). By GC-MS and multiple ion monitoring, we have shown the presence of 6-keto-PGF1 alpha (the stable metabolite of PGI2) in the umbilical artery incubation medium, while no trace of 6-keto-PGF1 alpha could be found in the placental medium. No conversion of AA to 6-keto-PGF1 alpha by placental microsomes was observed, even in the presence of antioxidants. The placenta possesses, in addition to the known 15-OH-PGDH and delta-13 reductase activities, a weak 9 OH PGDH which is specific for PGF2 alpha (and not PGI2 nor 6-keto-PGF1 alpha). GC-MS analysis showed that the expected metabolites of PGI2 through those three enzymes were not found in the placental medium, indicating that neither PGI2 synthesis nor metabolism could be demonstrated in the placenta. PMID- 6298906 TI - In vitro and in vivo ACTH-releasing activity of ovine CRF, sauvagine and urotensin I. AB - The ability of three homologous peptides, ovine corticotropin-releasing factor (CRF), sauvagine and urotensin I, to release ACTH was examined in vitro and in vivo. All three peptides exhibit statistically equivalent potencies to stimulate ACTH secretion both by cultured pituitary cells and in pharmacology blocked rats. These results suggest that all three peptides evolved from a common mammalian precursor that possessed high hypophysiotropic potency. PMID- 6298907 TI - [Echographic diagnosis of neuroblastoma in children]. AB - Ultrasounds have a high diagnostic value mainly in the structural assessment of the neoplasia. They are also very useful for the evaluation of the spread. In this paper the authors analyze the criteria for a differentiated diagnosis of Wilms' tumour. PMID- 6298908 TI - [Iatrogenic changes in the dural sac. Radiologic semeiology]. AB - The authors have studied the pathological appearances of follow-up and post operative radiculography. It has been possible to distinguish between lesions due to arachnoiditis and periduritis. PMID- 6298909 TI - [Molded milk duct calcifications in a case of ductal epithelioma of the breast]. PMID- 6298910 TI - [Cancer of the pancreas. Clinical review of 56 cases]. PMID- 6298911 TI - [Emphysematous cholecystitis. Microbiological study of a case]. PMID- 6298912 TI - The use of the biconical Challier-Laveissiere trap impregnated with Deltamethrine against Glossina. PMID- 6298913 TI - Notes on Culicoides (diptera, Ceratopogonidae) from the Sudan in relation to the epidemiology of bluetongue virus disease. PMID- 6298914 TI - [Esophageal granular cell myoblastoma associated with a gastric adenocarcinoma. A new case]. PMID- 6298915 TI - Effects of induced hypoprolactinaemia in the ram: plasma gonadotrophin levels, LH and FSH receptors and histology of the testis. AB - In order to investigate the effects of induced hypoprolactinaemia on gonadotrophin levels and testicular function in the ram, CB 154 was administered either in summer under natural photoperiod or in winter combined with a light regime stimulating prolactin release. Under natural photoperiod in summer, plasma FSH levels increased from 2.5 to 4 ng/ml in controls and from 2.5 to 8 ng/ml in CB 154-treated rams. Control and treated groups differed significantly (P less than 0.05) from the second week of treatment onwards. In the animals photostimulated in winter, treatment with CB 154 did not change plasma FSH levels; no effect of CB 154 on plasma LH levels was detected in either experiment. Treatment with CB 154 in summer led to an increase in the volumes of peritubular blood vessels and interstitial tissue. The number of Leydig cells per testis was significantly (P less than 0.05) increased in CB 154-treated rams. However, at the time of measurement, there was no change in the structure of the seminiferous tubules or in the production of germ cells. In winter in photostimulated animals, no effect of treatment with CB 154 was detected either on the structure of intestitial and tubular compartments or on spermatogenesis. The change in prolactin levels affected neither the number of LH receptors, expressed per Leydig cell, nor the number of FSH receptors, expressed per Sertoli cell, in the experiments. It is concluded that induced hypoprolactinaemia can modify the release of FSH in the ram and change the structure of the intertubular tissue but that it does not affect the number of LH and FSH receptors or impair spermatogenesis. PMID- 6298916 TI - A comparison of diethyldithiocarbamate and EDTA as antidotes for acute cadmium intoxication. AB - For mice which have been given the cadmium equivalent of CdCl2 X 2 1/2H2O at a level of 10 mg/kg, diethyldithiocarbamate (DDTC) administration at 500 mg/kg leads to a higher survival rate than is found with Na2CaEDTA under identical conditions. This occurs when the interval between administration of the cadmium salt and the antidote is either one or two hours. The surviving animals, however, are found to have appreciable levels of cadmium in their brain, liver and kidney when they were treated with DDTC, though their behavior appeared to be normal. The use of 50 mg/kg of DDTC after 2 hr in such cadmium poisoned mice also leads to a higher survival rate and reduced amounts of cadmium retained in the brain, liver and kidney. Unlike the other chelating agents which have been reported as useful in acute cadmium intoxication, DDTC appears to act by forming a lipid soluble complex which is largely immobilized in lipid containing tissues of the animal's body. PMID- 6298917 TI - Effect of glucose-6-phosphate dehydrogenase deficiency on the benz(a)pyrene toxicity for in vitro cultured human skin fibroblasts. AB - Human skin fibroblasts, isolated in vitro, from donors carrying the Mediterranean variant of glucose-6-phosphate dehydrogenase, exhibit a sharp decrease of hexose monophosphate shunt and NADPH/NADP+ ratio when compared to the fibroblasts from normal donors. This behavior is coupled to an increase of the resistance to cell death and growth inhibition induced by benz(a)pyrene, whose activation proceeds through the NADPH-dependent arene oxide formation. No differences were observed in the toxic effects of methylnitrosourea, a carcinogen that does not need metabolic activation, on normal and variant fibroblasts. PMID- 6298918 TI - Taste bud adenosine -3'5'-monophosphate phosphodiesterase: activity, subcellular distribution and kinetic parameters. AB - Higher activity of cyclic adenosine 3',5'-monophosphate (cAMP) phosphodiesterase (PDE) was found in homogenates from bovine circumvellate papillae bearing taste buds compared to activity in homogenates from areas surrounding these papillae in which no taste buds were present. With progressive purification of these homogenates cAMP PDE activity increased in the taste bud enriched fractions relative to that measured in the non-taste bud bearing epithelial tissue. The highest levels of cAMP PDE activity were measured in those taste bud fractions in which purification was greatest. Kinetic studies in both taste bud derived and control tissues showed two apparent Km values, one relatively high, the other, lower. cAMP PDE activity of taste bud membranes was enhanced by Mg++, Mn++ and imidazole and inhibited by ethylene-bis (beta-aminoethylether) N,N-tetra-acetic acid (EGTA), isobutyl methyl xanthine (IBMX), theophylline, and cyclic guanosine 3',5'-monophosphate (cGMP). The possible role for cAMP PDE in taste function is discussed. PMID- 6298919 TI - Inert tracer gas washout from mixed venous blood: the sloping alveolar plateau. AB - The aim of this model study was to investigate the mechanisms underlying the sloping alveolar plateau for inert tracer gases supplied to the lung by mixed venous blood. Transpulmonary gas exchange was simulated in an asymmetric lung model for conditions at rest and in exercise. For highly soluble gases, the calculations show that the varying amount of tracer gas dissolved in superficial parenchymal tissue and capillary blood causes a sustained stratification in the acinus during expiration and that this is mainly responsible for the slope. For this type of tracer gas, the slope is almost independent of variations in the molecular diffusion coefficient (D) of the gases. In contrast, for poorly soluble gases, the contributions of local parallel inhomogeneities of gas concentrations in the acinus and the continued gas exchange across the alveolo-capillary membrane are mainly responsible for the slope. The first factor, which depends on the asymmetric branching pattern of intra-acinar airways, increases with decreasing D values. The contribution of continued gas exchange to the slope is most pronounced under exercise conditions. This contribution is almost independent of the blood/gas partition coefficient, lambda, for lambda values less than 4.0. PMID- 6298920 TI - [Bone scintigraphy in hip pathology]. PMID- 6298921 TI - Monoclonal antibodies of hybridomas. AB - Among the most prominent biological advances of this century, the hybridoma technology stands out as providing the best tool for the analysis of complex antigens of pathogens and tumor cells. This technology will play a major role in the immunodiagnosis and treatment of infectious and malignant diseases. This review focuses on the most recent developments (1980-1981), with emphasis on human immunology. PMID- 6298922 TI - Membrane transport of anions across epithelia of mammalian small intestine and kidney proximal tubule. PMID- 6298923 TI - Endorphin related peptides. PMID- 6298924 TI - Properties of rheumatoid and normal synovial tissue in vitro and cells derived from them. Production of prostaglandins and collagenase in response to factors derived from cultured blood mononuclear cells and from synovium. PMID- 6298925 TI - [Ankylosing spondylarthritis]. PMID- 6298926 TI - Polyneuropathy with neuromyotonia elements. Anatomo-clinical, biochemical and electrographic study of two familial cases. PMID- 6298927 TI - [Stimulation of immunoreactive beta-endorphin with hypoglycemia and physical exertion]. PMID- 6298928 TI - Treatment of diverticular disease. PMID- 6298929 TI - Other causes of dyspepsia--especially abdominal pain of spinal origin. PMID- 6298930 TI - Relation between hypercalciuria and vitamin D3-status in patients with urolithiasis. AB - Patients with urolithiasis were divided into two groups, one (n = 38) with a urinary excretion of calcium exceeding 6.0 mmol/24 h and one (n = 32) with a calcium excretion lower than 6.1 mmol/24 h. The group of patients with a high urinary excretion of calcium had a significantly higher level of 25-hydroxy vitamin D3 (26.2 +/- 1.6 ng/ml) than had the group of patients with a normal urinary excretion of calcium (17.6 +/- 0.9 ng/ml) (p less than 0.001). A highly specific and accurate method, based on isotope dilution--mass spectrometry was used in the assay of 25-hydroxy vitamin D3. There was no over-all correlation between level of 25-hydroxy vitamin D3 and serum level of calcium (r = 0.1). The results are in accordance with the contention that the vitamin D3-status might be of some importance for the development of hypercalciuria in these patients. PMID- 6298931 TI - Chronic progressive external ophthalmoplegia. Clinical, electrophysiological, histochemical and ultrastructural studies of 14 cases. AB - Clinical, histochemical and ultrastructural findings concerning 14 cases with diagnosis of Chronic Progressive External Ophthalmoplegia are described. According to the clinical features the patients have been rated in two groups: the first including subjects with isolated ptosis or ptosis with external ophthalmoplegia and the second including subjects with a spreading of the muscular deficit and involvement of the neck and limbs. The most frequent histological and histochemical features are type I fibre atrophy, ragged-red fibres, DPNH-diaphorase reaction disorders and abnormal accumulation of lipids into the fibres. Electron microscopy reveal myofibrillar disorganization and clusters of polymorphous, abnormal mitochondria. In five cases mitochondria contain a variety of crystalline inclusions. Correlations between clinical data and histochemical and ultrastructural findings are discussed. Mitochondrial abnormalities are postulated to be a characteristic physiopathological pattern in CPEO. PMID- 6298932 TI - [Malignant fibrous histiocytoma. An attempt at a histo-clinical correlation in 21 cases]. AB - The malignant fibrous histiocytomas reported to the Department of Pathology of the University Cantonal Hospital of Geneva between 1971 and 1978 are reviewed and a clinico-histological study is conducted in 21 out of 27 cases. An attempt is made to correlate some histological features (i.e. mitoses, atypical figures, inflammatory cells, necrotic areas, stroma, cellular morphology) with clinical course (i.e. local recurrence and metastases). No clinico-histological correlation was found and the result is thus comparable to the literature, which suggests that factors influencing prognosis are size, localization and depth of the tumor: the prognosis of small, distal and superficial tumors is better. Furthermore, using the peroxidase bridge technique for lysozyme, 3 positive reactions were found in 17 assays. The authors' immunohistochemical findings together with other histological features of the tumors and results reported in the literature suggest that the malignant fibrous histiocytoma is derived from an undifferentiated mesenchymal cell rather than from "histiocytes" as its name would suggest. PMID- 6298933 TI - [Treatment of reflux esophagitis with ranitidine. A multicentric prospective study]. AB - Thirty-eight patients with erosive-ulcerative reflux esophagitis were treated in an open trial with the histamine-H2-receptor antagonist ranitidine (150 mg twice daily). At endoscopy after 6 weeks there was evidence of complete healing of all epithelial defects in 20 of 38 patients (53%). Continuation of treatment for another 6 weeks in 13 patients was followed by healing in 6 additional patients as judged by endoscopy. Although the symptoms of the group as a whole improved significantly during treatment, there was no correlation between the degree of symptomatic improvement and healing of esophagitis. Also, the healing rate did not depend on age, sex, smoking and drinking habits or on the severity of esophagitis. It is concluded that medical treatment of reflux esophagitis with ranitidine is promising, even in severe cases previously considered candidates for surgery. PMID- 6298934 TI - [Evaluation of the treatment of symptomatic urinary tract infection with a single dose of antibiotics or bicarbonate]. AB - 68 women with acute, uncomplicated urinary tract infection confirmed bacteriologically were randomized to receive either a single oral dose of cotrimoxazole, a single oral dose of amoxycillin, or an oral dose of Na bicarbonate. Results comparable to previous reports were obtained with the cotrimoxazole regimen. On the other hand, the efficacy of single-dose amoxycillin appeared to be inferior. It is possible that pharmacocinetic differences influence the efficacy of these drugs. PMID- 6298935 TI - Infection of normal human epithelial cells by Epstein-Barr virus. AB - Primary cultures of epithelial cells were grown from the tonsils and adenoids of patients with diseases not related to Epstein-Barr virus. The cells could not be infected by Epstein-Barr virus. Fluorescein-labeled Epstein-Barr virus and a cytofluorograph were then used to show that the epithelial cells do not have detectable receptors for the virus. However, implantation with Epstein-Barr virus receptors gave the cells the ability to bind the labeled virus. One to 5 percent of receptor-implanted cells exposed to the transforming B95-8 substrain of the virus expressed Epstein-Barr nuclear antigen. The early and viral capsid Epstein Barr virus-determined antigens were not detected in the virus-infected cultures. The results show that normal human epithelial cells from the nasopharynx become susceptible to infection by Epstein-Barr virus when the membrane barrier resulting from the lack of viral receptors is overcome by receptor implantation. PMID- 6298936 TI - Demonstration of a saturable binding site for thyrotropin in Yersinia enterocolitica. AB - Several lines of evidence suggest that there might be immunologic cross reactivity between the thyroid plasma membrane in humans and antigenic determinants in the enteric pathogen Yersinia enterocolitica. Studies were therefore performed to determine whether Y. enterocolitica, like the thyroid membrane, contains a thyrotropin binding site. A saturable binding site for bovine thyrotropin was indeed demonstrable, particularly in preparations of the organism that have been treated with ethylenediaminetetraacetate and lysozyme. Hormonal specificity of the binding site, as judged from the inhibition of binding of 125I-labeled bovine thyrotropin, was similar to that of the thyrotropin receptor in human thyroid tissue. PMID- 6298937 TI - Protons and anaerobiosis. AB - During oxygen limitation in animals, glucose can be fermented via several metabolic pathways varying in energetic efficiency and leading to various end products (such as lactate, alanopine, octopine, succinate, or propionate). Because of opposite pH dependencies of proton production by fermentation and by hydrolysis of adenosine triphosphate formed in the fermentation, the total number of moles of protons generated is always two per mole of the fermentable substrate. However, two and three times more adenosine triphosphate can be turned over per mole of protons produced in succinate and propionate fermentations, respectively, than in lactate fermentation. PMID- 6298938 TI - Tolerance develops to the disruptive effects of delta 9-tetrahydrocannabinol on primate menstrual cycle. AB - Long-term exposure of sexually mature female rhesus monkeys (Macaca mulata) to thrice weekly injections of delta 9-tetrahydrocannabinol resulted in a disruption of menstrual cycles that lasted for several months. This period was marked by an absence of ovulation and decreased basal concentrations of gonadotropin and sex steroids in the plasma. After this period, normal cycles and hormone concentrations were reestablished. These studies demonstrate that in rhesus monkeys subjected to long-term treatment with delta 9-tetrahydrocannabinol tolerance develops to the disruptive effects of the drug on the menstrual cycle. PMID- 6298939 TI - Adenylate cyclase activation shifts the phase of a circadian pacemaker. AB - Forskolin, a highly specific activator of adenylate cyclase, produced both delay and advance phase shifts of the circadian rhythm recorded from the isolated eye of the marine mollusk Aplysia. The phase dependence of the response to forskolin was identical to that with serotonin, which also stimulates adenylate cyclase in the eye. The ability of agents to activate adenylate cyclase in homogenates was correlated with their ability to shift the phase of the circadian oscillator. These results along with earlier findings show that adenosine 3',5'-monophosphate mediates the effect of serotonin on the rhythm and regulates the phase of the circadian pacemaker in the eye of Aplysia. PMID- 6298940 TI - Bidirectional transmission at the rectifying electrotonic synapse: a voltage dependent process. AB - Rectifying properties of electrotonic synapses established by the crayfish giant motor fiber are associated with a more negative resting membrane potential in the presynaptic than in the postsynaptic side of the junction. An increased junctional conductance and bidirectional transmission are produced, with almost no delay, by inverting this polarization. PMID- 6298941 TI - A functional role for an opiate system in snail thermal behavior. AB - The terrestrial snail Cepaea nemoralis, when placed on a 40 degrees C hot plate, lifts the anterior portion of its foot. The latency of this response is influenced by morphine and by naloxone in a dose-dependent and time-dependent manner. Morphine increases the time taken to respond, whereas naloxone reduces it. Furthermore, naloxone abolishes the effect of morphine. These results indicate that an opiate system may have a role in this behavior, which resembles that reported in vertebrates. PMID- 6298942 TI - Swine influenza-like viruses in turkeys: potential source of virus for humans? AB - Influenza A viruses (subtype H1N1), recently isolated from turkeys in different areas of the United States, were determined to be closely related to strains typically associated with pigs. This conclusion was based on comparisons of H1N1 isolates from pigs, humans, ducks, and turkeys with polyclonal and monoclonal antibodies, RNA-RNA competitive hybridization, and replication studies. One of the H1N1 isolates from turkeys caused influenza in a laboratory technician, who displayed fever, respiratory illness, virus shedding, and seroconversion. These results suggest that turkeys as well as pigs are involved in the maintenance of these viruses and their transmission to humans. PMID- 6298943 TI - Chemotherapy of lung cancer. PMID- 6298944 TI - Single-dose ifosfamide: efficacy studies in non-small cell lung cancer. PMID- 6298945 TI - Ifosfamide in the treatment of extensive non-oat cell carcinoma of the lung. PMID- 6298947 TI - Efficacy of technetium Tc 99m pyrophosphate imaging in patients with equivocal myocardial infarction. AB - We studied the efficacy of technetium Tc 99m pyrophosphate imaging in patients with equivocal evidence of acute myocardial infarction. Only patients with positive enzyme findings (regardless of ECG findings) had scans with greater than or equal to 2+ focal uptake. None of 26 patients with negative or equivocal enzyme findings (regardless of ECG findings) had greater than 2+ diffuse uptake. These results support the contention that infarct-avid imaging has little clinical utility in patients with equivocal evidence of myocardial infarction. PMID- 6298946 TI - Use and safety of high-dose ifosfamide. PMID- 6298948 TI - Enterohepatic fistula associated with amebic liver abscess. PMID- 6298949 TI - Rotavirus screening in adult cancer patients. PMID- 6298950 TI - Current concepts on etiology and pathogenesis of diarrhoea caused by viruses. PMID- 6298951 TI - [Our contribution to the diagnosis of glomus jugulare tumors]. PMID- 6298952 TI - [Polyneuropathy in a patient with acromegaly]. PMID- 6298953 TI - [Neuropathy as a result of cryptogenic tetany in 2 patients]. PMID- 6298954 TI - The reversibility of cancer. PMID- 6298955 TI - The effect of long-term high-fibre diets in diabetic outpatients. AB - Diets containing large amounts of dietary fibre have been shown to be beneficial in improving diabetic control. We investigated the practical aspects of administering a high-fibre diet to diabetic outpatients in Cape Town, using readily available, low-cost foodstuffs with a high dietary fibre content. Ten patients were followed up over a period of 9 months, for 3 months of which a high fibre diet was prescribed. Although only 3 patients approached the projected dietary fibre intake, significant negative correlations were found between the mean plasma glucose changes and the dietary fibre increments (r = 0,704; P less than 0,05) and between the mean serum triglyceride changes and the dietary fibre increments (r = -0,741; P less than 0,05). These findings suggest that, were it not for poor dietary compliance, a high-fibre diet might result in significant improvement in diabetic control, and that education and motivation are of prime importance when making major changes to patients' eating habits. PMID- 6298956 TI - Chikungunya virus infection. A retrospective study of 107 cases. AB - A retrospective study of 107 cases of serologically proven chikungunya (CHIK) virus infection was undertaken. All respondents had contracted the disease at least 3 years previously; 87,9% had fully recovered, 3,7% experienced only occasional stiffness or mild discomfort, 2,8% had persistent residual joint stiffness but no pain, while 5,6% had persistent joint pain and stiffness and frequent effusions. Synovial fluid from 3 patients was analysed. All the patients with persistent joint pain and stiffness had very high antibody titres against CHIK virus. PMID- 6298957 TI - The Eaton-Lambert syndrome. AB - The Eaton-Lambert syndrome is a non-metastatic manifestation of oat-cell carcinoma of the bronchus, although it has been reported in patients with carcinoma at other sites. The clinical picture is usually one of subacute muscular fatiguability with weakness and wasting affecting the proximal parts of the limbs and trunk, but occasionally the external ocular and bulbar muscles are involved. The weakness is often myasthenic, but it has been observed that muscle power may in fact increase after brief exercise. The pathogenesis, diagnosis, electrophysiological features and some aspects of therapy are reviewed. PMID- 6298958 TI - Severe chronic constipation as a surgical problem. PMID- 6298959 TI - Lymphoscintigraphy in the staging of solid tumors. PMID- 6298960 TI - Evolution of studies on antireceptor antibodies and disease. PMID- 6298962 TI - Autoantibodies to hormone receptors. PMID- 6298961 TI - The thyrotrophin receptor. PMID- 6298964 TI - Plasma beta-endorphin immunoreactivity in dogs during anesthesia, surgery, Escherichia coli sepsis, and naloxone therapy. AB - To improve understanding of the role of endorphins in septic shock, we examined the effects of anesthesia, splenectomy, live Escherichia coli infusion, and treatment with naloxone, respectively, on plasma beta-endorphin immunoreactivity (beta-EI) and plasma cortisol in dogs. Baseline levels of plasma beta-EI and cortisol were established in awake dogs. Pentobarbital anesthesia alone did not affect plasma beta-EI, but splenectomy was followed by a significant (P less than 0.001) rise in both plasma beta-EI and cortisol. Infusion of saline over a 3-hour period following splenectomy induced no further increase in plasma beta-EI, but infusion of live E. coli in splenectomized dogs caused a further rise in plasma beta-EI (P less than 0.02). Following induction of septic shock in a separate group of splenectomized animals, treatment with naloxone (3 mg/kg bolus and 2 mg/kg/hr infusion) did not alter the rise in plasma beta-EI. These results confirm release of beta-endorphin during septic shock and further implicate the hypothalamic-pituitary-adrenal axis in its pathophysiology. Based on the finding that naloxone did not affect the dynamics of plasma beta-EI, mechanisms are postulated to explain the therapeutic value of this drug in septic shock. PMID- 6298965 TI - Hemorrhagic cytomegalovirus colitis in a patient with uremia. PMID- 6298966 TI - Hemidiaphragmatic paresis after cervical herpes zoster. PMID- 6298967 TI - The effect of a medicinal Chinese herb on platelet function. AB - We investigated the effect of the Chinese herb Injectio Salvia Miltiorrhizae (ISM) on human platelet function in vitro. ISM inhibited platelet aggregation and serotonin release induced by either ADP or epinephrine in a dose dependent manner. This effect of ISM was observed with both gel-filtered platelets (ID50 = 8-30 micrograms ISM/ml gel-filtered platelets) and platelets in plasma (ID50 = 400-900 micrograms ISM/ml of platelet-rich plasma). The active molecule(s) in ISM was heat stable, resistant to acid, base and proteolysis and fractionated on Sephadex 6-25 at MW approximately 280. ISM did not interact with the platelet alpha-adrenergic receptor, but increased cAMP in intact platelets. The results are consistent with the concept that ISM inhibition of platelet aggregation and release is mediated by an increase in platelet cAMP. The exact mechanism whereby ISM increases platelet cAMP appears to be that of inhibition of cyclic AMP phosphodiesterase. The effect of ISM on platelet function is one mechanism which might explain the therapeutic effect of ISM in experimental and clinical coronary artery disease. PMID- 6298963 TI - Biological effects of antibodies to lymphocyte surface receptors. PMID- 6298968 TI - Decrease of platelet activity after intake of small amounts of eicosapentaenoic acid in diabetics. PMID- 6298969 TI - 1,25-Dihydroxyvitamin D production stimulated by dibutyryl 3', 5'-cyclic AMP in normal subjects and a patient with pseudohypoparathyroidism. PMID- 6298970 TI - Fibronectin in human hepatocellular carcinoma. AB - Human liver specimens with hepatocellular carcinoma were studied for distribution of fibronectin by immunofluorescence staining. Fibronectin was detected in the cytoplasm and/or on the cell surface in certain cases, which is in contrast to the observations in vivo for hepatocytes of normal and other fibrotic livers. The present finding is also in contrast to those for various other carcinomas so far reported. PMID- 6298971 TI - Recent developments with copper proteins. PMID- 6298972 TI - Semiquinone formation in flavo- and metalloflavoproteins. PMID- 6298973 TI - The metabolism of superoxide anion and its progeny in blood cells. PMID- 6298974 TI - The flavin redox-system and its biological function. PMID- 6298975 TI - Protection from paraquat-induced lung damage and lethality in adult rats pretreated with clofibrate. PMID- 6298976 TI - The influence of sulfide and cyanide on axonal function. AB - The absorption of hydrogen sulfide (H2S) gas by sciatic nerve bundles from Rana pipiens produces an anaesthetic effect of short duration. Subsequent compound action potential (CAP) levels are higher than before exposure while the conduction velocity (CV) shows only small variation. Exposure to hydrogen cyanide (HCN) gas produces declining CAP and CV levels. The effect of H2S on the CAP is faster than that of HCN and the recovery time is shorter, although there is evidence of permanent change. Unlike HCN there is no indication that H2S inhibits the energy metabolism of the nerve cells, except possibly when the nerves are exposed to extremely high sulfide concentrations. A mode of action for H2S is proposed. PMID- 6298977 TI - Actions of nortetrodotoxin on frog muscle and squid axon. AB - Nortetrodotoxin, prepared by oxidation of the C6 end of tetrodotoxin, probably exists in solution as an equilibrium mixture of a hemilactal and a lactone form of a hydrated ketone of C6. On both frog muscle fibers under constant-current conditions and squid giant axon under voltage-clamped conditions, nortetrodotoxin selectively blocks the sodium channel. On frog muscle fibers, the solution of nortetrodotoxin used, which was several months old, was 1/13 as active as tetrodotoxin. On the squid giant axon, a freshly prepared solution of nortetrodotoxin was about 1/4 as active. These observations indicate that nortetrodotoxin possesses a substantial degree of biological activity. The suitability of nortetrodotoxin as an intermediate for synthesizing derivatives of tetrodotoxin is discussed. PMID- 6298978 TI - The effects of a few xenobiotics on certain phosphatases in the plasma of Clarias batrachus and Cirrhina mrigala. AB - The effects of sublethal concentrations 1/2, 1/4 and 1/8 fractions of 96-h LC50 of aldrin and Swascofix CD-38 on alkaline, acid and glucose-6-phosphatases in the plasma of Clarias batrachus and Cirrhina mrigala were studied. These enzymes increased after exposure of fish to both xenobiotics. The increase was maximal after 20 days exposure to the highest concentrations. The enzyme increase was greatest in the fish exposed to aldrin. PMID- 6298979 TI - Adsorption of nucleic acids on asbestos fibers in vitro. AB - Adsorption of DNA and RNA, accompanied by liberation of orthophosphate, occurred on contact with chrysotile asbestos fibers. This indicated that interaction of macromolecules and their components may be a factor, possibly involved in the manifestation of the toxic effect of asbestos fibers. PMID- 6298980 TI - Mechanism of haemolysis by chrysotile fibres. AB - Haemolysis by chrysotile fibres may be related to an adsorption of the red blood cell (RBC) membranes on the fibres and not to an interaction between magnesium (Mg) from the fibres and sialic acid from RBC. This was demonstrated using neuraminidase (NANASE)-treated RBC and Mg-depleted chrysotile fibres. The mechanism may be extended to other minerals since a relationship can be obtained between the ability of various minerals to adsorb liposomes of dipalmitoyl phosphatidylcholine (DPPC) and their haemolytic activity. PMID- 6298981 TI - Primary hepatocellular carcinoma and hepatitis B virus. PMID- 6298982 TI - A 12-month surveillance of 'flu-like infections in a sample from the population of Lagos. AB - 5,576 people who had two or more of the following symptoms, running nose, cough, fever and crepitations/rhonchi over the chest were interviewed. 1,355 specimens of nasal and throat swabs from those interviewed were processed for viruses associated with upper respiratory tract infection. There was a sharp rise in 'flu like symptoms in March 1976, followed by a much higher incidence from June, reaching a peak in August. Influenza A was isolated from most of the specimens collected. A few para-influenza II were also isolated. Our findings show that severe outbreaks of acute viral infection of the respiratory tract occurred during our wet season particularly around August. PMID- 6298983 TI - [Effect of estradiol on 3',5'-AMP-phosphodiesterase activity in rat uterus. Participation of hormone receptors, the role of guanyl nucleotides]. AB - Estradiol (10(-7)-2 . 10(-5)M) is shown to inhibit the activity of phosphodiesterase of the soluble fraction of the mature rat womb tissue and to have no effect on the activity of adenylate cyclase of this tissue membrane fraction and on the phosphodiesterase of the tissue of the brain, heart and outer segments of the rod-cell. When blocking the estradiol cytosol receptors by clomiphene there is no inhibitory effect of the hormone on the activity of phosphodiesterase of the womb tissue preparations. Specific binding of estradiol by the cytosol receptors increases in the presence of guanylic nucleotides (10( 5)-10(-4)M); ATP does not affect this process. In the presence of guanyl-5 ilimidodiphosphate (Gpp(NH)p), a nonhydrolyzed analog of GTP, clomiphene does not block receptor binding of estradiol. ATP and GTP (10(-6)-10(-5)M), contrary to Gpp (NH)p, inhibit the phosphodiesterase activity. Independent of their effect on the enzymic activity, all the studied nucleotides partially or completely eliminate the inhibitory effect of estradiol on phosphodiesterase. PMID- 6298984 TI - Effect of pulsed ultrasound on human erythrocytes in vitro. AB - Pulsed echo sonication of whole blood affects the O2-Hb dissociation curve and produces modifications of the erythrocyte membrane, whereas Hb remain unchanged. After sonication ATP-ase (Na+K+ dependent) activity and agglutination disappear. Sonication detaches erythrocytes-membrane antigens which are found in the supernatant, this effect is reversible. AB antigens detached by sonication were transferred to sonicated O group erythrocytes. Examination by transmission electron microscopy suggested that the glycoproteic cup or glycocalyx of the red cells was removed as a result of sonication. PMID- 6298985 TI - Progress of peripheral uremic neuropathy during continuous ambulatory peritoneal dialysis (CAPD). PMID- 6298986 TI - Isolation of a new serotype of infectious bronchitis-like virus from chickens in England. PMID- 6298987 TI - Vaccines for EHV1. PMID- 6298988 TI - Organophosphate poisoning: modifications in acid base equilibrium and use of sodium bicarbonate as an aid in the treatment of toxicity in dogs. AB - We injected technical quality DDVP ip into 13 dogs at a dose of 30 mg/kg, a dose 100% lethal in 27 minutes. We monitored blood pH, arterial gases, and cholinesterase activity before and during the intoxication and found acidosis. We used iv sodium bicarbonate and achieved recovery of acid-base balance, improvement in the poisoned state, and survival of 84.6% of the patients. PMID- 6298989 TI - Ultrastructural changes and antigen localization in tissues from foot-and-mouth disease virus-infected guinea-pigs. AB - Foot-and-mouth disease virus (FMDV)-induced ultrastructural changes in guinea-pig tongue, heelpad, mammary and liver tissues were examined using scanning and transmission electron microscopy. FMDV infection caused cell rounding and the release of virus in membrane limited vesicles in the animal tissues similar to that seen in other work in cell cultures. Microfilaments were present which may be responsible for cell rounding. Immunoperoxidase labeling revealed the attachment of the virus-infection associated (VIA) antigen to the smooth vacuoles of mammary and liver tissues, and to milk fat globules. The electron microscope immunoperoxidase procedure increased the sensitivity of detection sufficiently to allow the visualization of VIA antigen in tissues not previously shown to have the antigen. It is postulated that the release of the smooth vacuoles from the liver cells stimulates the animal's immune response to the VIA antigen. PMID- 6298990 TI - Comparative studies on the growth of Australian bluetongue virus serotypes in continuous cell lines and embryonated chicken eggs. AB - The replication of cell culture passaged Australian bluetongue virus (BTV) isolates, serotypes 20 (CSIRO19) and 1 (CSIRO156), and an untyped BTV (CSIRO154) was assessed in eight continuous cell lines (one derived from baby hamster kidney cells, BHK-21; three derived from monkey kidney cells, Vero, LLC-MK2 and CV-1P; a foetal ovine lung and a mouse fibroblast cell line, CSL503 and L929, respectively, a Super-Vero-Porcine stable cell line, SVP; and a mosquito cell line, Aedes albopictus cells) and in 11-day-old embryonated chicken eggs (ECE) at different multiplicities of infection. All three viruses replicated in the cell lines tested, maximum extracellular virus yields being attained from BHK-21 cells at high multiplicities of infection (approximately 10 PFU per cell). Also BHK-21 cells produced much higher yields of virus than the other cell lines tested when low multiplicities of infection were used (approximately 10(-4) PFU per cell). All BTV serotypes multiplied in Singh's Aedes albopictus cells with no cytopathogenic effects over the 4 day period tested. The viruses also replicated in 11-day-old ECE; however, the sensitivity of ECE for growth of the Australian serotypes was not as high as has been reported for BTV isolates in other countries. In all cell culture systems and in ECE, BTV1 and BTV20 replicated more efficiently than did CSIRO154 virus. PMID- 6298991 TI - Role of the spleen in the pathogenesis of experimentally induced bovine leukemia virus infection. AB - In an extension of a previous pathogenesis study, bone marrow and other tissues from four experimentally inoculated cattle were tested for virus between the 13th and 20th days after experimental inoculation with bovine leukemia virus. BLV was detected in the blood of three, spleen of two, lymph node of two and bone marrow of only one of the inoculated cattle. In additional studies, four splenectomized and two intact control calves were also examined. Two of these calves were splenectomized before BLV inoculation and two after a persistent virus infection had been established. Results indicated that the removal of the spleen affected neither the establishment and persistence of virus infection nor the development and maintenance of serological responses to viral antigens. PMID- 6298992 TI - Studies of enteric coronaviruses in a feline cell line. AB - Development is reported of a feline cell line which can support the growth of coronaviruses from canine (CCV), feline (FIPV) and porcine (TGEV) species. The cell culture has been serially transferred over 100 times and has retained its initial growth requirements, proliferative capacity and morphologic features. Each virus had specific growth characteristics in this cell culture although all produced a similar CPE and plaques under agar. Cross neutralization studies demonstrated a two-way relationship between TGEV and CCV and between TGEV and FIPV, whereas a one-way relationship was demonstrated between CCV and FIPV. PMID- 6298993 TI - Biochemical characterization of staphylococci isolated from goats. AB - Caprine strains of staphylococci show strong and early results in carbohydrate fermentation tests. Their ability to produce DNase and acetoin is weak but they possess a reasonably high degree of phosphatase activity. All the 82 isolates of Staphylococcus aureus isolated from clinically healthy and sick goats produced acid from D-(+)-glucose aerobically at 37 degrees C within 24 h, 98.8% fermented both D-mannitol and maltose, 98.7% fermented sucrose, and 81.7% fermented D-(+) xylose with acid production, but none fermented L-sorbose. Of the ten coagulase negative staphylococci characterized to be of animal origin, three produced DNase, but none fermented dulcitol and L-sorbose. All fermented D-(+)-glucose, maltose and D-mannitol, eight fermented D-(+)-xylose and D-sorbitol with acid production. Whilst other coagulase-positive and coagulase-negative staphylococci showed different antibiotic susceptibility patterns, the ten coagulase-negative staphylococci characterized to be of animal origin showed similar antibiotic susceptibility patterns. PMID- 6298994 TI - Malignant rhabdoid tumor of the kidney. Histopathology, ultrastructure and comments on differential diagnosis. AB - Clinical and histopathological features of two cases of malignant rhabdoid tumor of the kidney are presented. One of these cases was also studied by electron microscopy. Histologically, both tumors consisted of an admixture of undifferentiated polygonal or elongated cells and cells with abundant eosinophilic cytoplasm frequently containing hyaline globular structures. Ultrastructurally, these cytoplasmic inclusions were composed of large masses of actin-size and intermediate-size filaments. The poor prognosis of this type of tumor is emphasized and histological criteria for differential diagnosis from other malignant renal tumors of childhood and adolescence are discussed. PMID- 6298995 TI - Effect of X-ray irradiation on primary mineralization in rat alveolar bone. AB - The effect of X-ray irradiation on the process of primary mineralization in bone was studied by biochemical and ultrastructural methods. A single dose of 1500R was administered to the head region of rats. The animals were examined immediately after irradiation and 1, 2 and 3 weeks later. Fractions of isolated cells and extracellular matrix vesicles were prepared from the maxillary alveolar bone of irradiated and untreated rats by collagenase digestion and differential centrifugation. The protein content and activities of vesicular phosphatases were determined in both fractions. A continuous decrease in the activity of alkaline phosphatase could be observed in both cell and matrix vesicle fractions during a three-week follow up after irradiation. Acid phosphatase activity decreased only in the vesicle fraction. Transmission electron microscopy of irradiated bone tissue revealed that many matrix vesicles were devoid of intact membranes and apatite crystals. Calcifying nodules were abundant in the matrix without their apparent fusion into larger mineralized structures. It is suggested that irradiation interferes with enzymatic processes associated with primary mineralization. PMID- 6298996 TI - Synovial sarcoma enzyme histochemistry of a typical case. AB - A typical case of biphasic synovial sarcoma was studied using enzyme histochemistry. A marked difference between the staining characteristics of the spindle cells and the epithelial-like cells was demonstrated by reactions for various hydrolytic enzymes. The epithelial-like cells exhibited a strong reactivity for alkaline phosphatase, acid phosphatase, adenosine triphosphatase and nonspecific esterase, whereas spindle-cells were completely unreactive when tested for these enzymes. This is, to our knowledge, the first report demonstrating differences in the enzymatic pattern of the two cell populations which compose synovial sarcoma. PMID- 6298997 TI - Glial fibrillary acidic protein in medulloblastomas and other embryonic CNS tumours of children. AB - Investigation of GFAP in 50 medulloblastomas showed a few GFAP-positive tumour cells in 5 cases only; 17 tumours were negative, and 28 showed a "pseudopositivity", i.e. GFAP-bearing cells were identified as reactive or degenerating astrocytes, intermingled with tumour elements. A high GFAP content was seen in 2 small-cell gliomas of the cerebellum, whereas 3 pineoblastomas, 2 neuroblastomas of CNS, and one medulloepithelioma were negative. GFAP is a very good method for identificating astrocytes, but does not seem to be reliable for identifying the origin of undifferentiated tumours such as medulloblastomas. In these neoplasms glial differentiation is lacking or extremely rare, GFAP positivity being mostly an artifact. The investigation of small tumour samples or the positivity of a single cell are inadequate data for a correct evaluation of the findings, especially taking in mind that GFAP of degenerated astrocytes can be phagocytised by cells other than glial (e.g., macrophages, epithelial and meningioma cells). The importance of carefully checking the whole structure of the tumour is stressed, GFAP positivity or negativity being not a sufficient criterion for its nosological classification. PMID- 6298998 TI - Primary renal tumours in the first year of life. A population based review. AB - Of 30 patients presenting with primary renal tumours in the first year of life, there were 23 Wilms' tumours (15 classical, six epithelial and two rhabdomyomatous), three rhabdoid neoplasms and four mesoblastic nephromas. Criteria for the diagnosis of rhabdoid tumours and mesoblastic nephromas are discussed with reference to histological difficulties. Although Wilms' tumour was the commonest neoplasm, mesoblastic nephroma predominated in the first three months of life. The clinical behaviour of the cases is reviewed, and rhabdoid tumours, although relatively few in number, accounted for a significant part of the overall mortality. PMID- 6298999 TI - Fatal lipid storage myopathy with deficiency of cytochrome-c-oxidase and carnitine. A contribution to the combined cytochemical-finestructural identification of cytochrome-c-oxidase in longterm frozen muscle. AB - Two newborn female siblings fell ill with apathy, failure of suckling and a generalized progressive muscular hypotonia. Death occured at the age of 7 weeks, obviously caused by impairment of respiratory musculature. Biochemical studies in one child revealed carnitine deficiency especially in skeletal muscle; hepatic encephalopathy was absent. Both children had a generalized hyperaminoaciduria, an unusual finding in primary carnitine deficiency. Besides fatty metamorphosis of the liver, bilateral hydroureters and tubular calcifications of both kidneys, morphological studies showed a generalized lipid storage myopathy which predominated in Type-I-fibres and was accentuated in the muscles of the neck. Enzymehistochemical electron microscopy in longterm frozen muscle demonstrated that cytochrome-c-oxidase activity was absent not only in myopathic but also in most of the morphological unchanged muscle fibres. Only some fibres and endothelial cells displayed normal activity of mitochondria. Biochemically no cytochrome aa3 (cytochrome-c-oxidase) could be found in skeletal muscle; cytochrome b was almost undetectable. --In newborns with fatal lipid storage myopathy and carnitine deficiency it seems necessary to look for additional defects in the respiratory chain. Enzyme histochemical electron microscopy is a sensitive method in identifying cytochrome-c-oxidase even after a 12 months period of storage. PMID- 6299000 TI - The pars tuberalis of the human pituitary. A histologic, immunohistochemical, ultrastructural and immunoelectron microscopic analysis. AB - Forty autopsy pituitaries were studied to elucidate the histology, immunocytology and ultrastructure of pars tuberalis in subjects with normal and abnormal endocrine homeostasis. Pars tuberalis consisted mainly of gonadotrophs interspersed with few corticotrophs and thyrotrophs, histologically resembling those of pars distalis. Somatotrophs and lactotrophs were not identified. There were no histologic differences attributable to age or sex. In cases of glucocorticoid excess, pars tuberalis corticotrophs showed Crooke's hyalinization. Following castration or hypophysectomy, pars tuberalis gonadotrophs exhibited more intense immunostaining for FSH and LH than did normals. Ultrastructural analysis revealed gonadotrophs and corticotrophs showing no evidence of active secretion; immunoelectron microscopy demonstrated FSH, LH and ACTH in secretory granules. By light microscopy, squamous nests, often identified in pars tuberalis, were positive for immunoreactive keratin; cells at their periphery contained FSH, LH or ACTH, indicating derivation of nests by squamous metaplasia from gonadotrophs and corticotrophs. By electron microscopy, clusters of epithelial cells containing desmosomes and tonofilaments were surrounded by granulated gonadotrophs. Human pars tuberalis cells represent mainly a subpopulation of gonadotrophs possessing all organelles required for synthesis and storage of hormones but showing ultrastructural features of functional inactivity; the reasons for this inactivity and for the formation of squamous nests remain unexplained. PMID- 6299002 TI - Enterovirus isolation from a case of hand, foot, and mouth disease. PMID- 6299001 TI - EPR and Mossbauer spectroscopy investigations on the metal ion contents of Sendai virus components. AB - EPR and Mossbauer spectroscopy indicate that Sendai virus contains iron ions in paramagnetic states. Spectral data show that the iron ions are in an oxidized form (Fe3+), having low and high spin states (S = 1/2 and S = 5/2). On enrichment of Sendai virus with 57Fe, the concentration of Fe3+ ions substantially increases in the virus preparations. The Fe3+ ions in the high spin state appear to be tightly bound to the virus components; they are not significantly removed by dialysis. The five main proteins separated by SDS gel electrophoresis from 57Fe enriched Sendai virus contain the signal corresponding to the presence of Fe3+ ions in the high spin state. The concentration of Fe3+ ions is, however, about five times higher in the HN polypeptide than in the other four components. It is suggested that Fe3+ (5/2) ions might be a structural component of the Sendai virus HN polypeptide. PMID- 6299003 TI - Free cottontail rabbit papillomavirus DNA persists in warts and carcinomas of infected rabbits and in cells in culture transformed with virus or viral DNA. AB - We have compared warts and carcinomas from cottontail and domestic rabbits for the presence of cottontail rabbit papillomavirus (CRPV) and the status of the viral DNA genome. Our studies indicate that benign warts from cottontail rabbits, whether found naturally or induced in the laboratory, contain large amounts of virus and on the average 1000 copies of the virus genome per cell. Both benign warts and carcinomas from domestic rabbits contain significantly reduced levels of virus relative to cottontail rabbit warts and an average of 100 copies of the virus genome per cell. A single sample of a naturally occurring cottontail rabbit carcinoma contained approximately 80 copies of the viral genome per cell. None of the tumors that we have analyzed thus far appear to have integrated viral genomes by Southern blot analysis of undigested and restriction endonuclease-digested DNA samples. Furthermore, the CRPV genome present in domestic rabbit carcinomas and a cottontail rabbit carcinoma appears identical by restriction endonuclease mapping to that present in papillomas of cottontail and domestic rabbits indicating that no major deletions or rearrangements of the CRPV genome had occurred during the progression of benign to malignant tumors nor was a variant of wild-type CRPV responsible for this phenomenon. Finally, we have demonstrated morphological transformation in vitro of NIH 3T3 and C127 cells upon infection with purified CRPV and upon transfection with purified CRPV DNA. Furthermore, single cell clones derived from transformed foci contain free forms of CRPV DNA that persist through continued passage in culture. Cells transformed by CRPV grow in soft agar in vitro and produce tumors in athymic nude mice. PMID- 6299004 TI - Isolation and mapping of cDNA hybridization probes specific for ecotropic and nonecotropic murine leukemia proviruses. AB - To study the structure of murine leukemia proviruses in AKR mice by Southern hybridization, we have isolated and mapped ecotropic AKV and nonecotropic MCF derived cDNA restriction fragments. The ecotropic-specific probes originate from four regions of the AKV genome which include the corresponding recombinant region of two MCF viruses (VI-36 and 247). We also isolated two nonecotropic probes from the recombinant region of MCF V1-36. The probes were characterized by (i) mapping of restriction fragments at the 3' end of AKV and MCF V1-36 by a two-dimensional gel strategy, (ii) hybridization of restriction fragments to the related viral RNA genomes followed by electrophoresis, (iii) two-dimensional fingerprinting of single-stranded restriction fragments, and (iv) DNA sequence analysis of the ecotropic probes. The ecotropic AKV and nonecotropic MCF probes discriminate between two populations of endogenous murine leukemia viruses and show that the MCF viruses are not present in the germ line of AKR mice. PMID- 6299005 TI - The genetic analysis of recombination using adenovirus overlapping terminal DNA fragments. AB - We have studied the consequences of genetic recombination between overlapping terminal fragments of adenovirus genomes with respect to markers in the overlapping sequence. The findings are consistent with general recombination occurring approximately isotonically within the interval. In particular, single markers within the overlap, scored nonselectively, showed frequencies of recovery dependent on the position of their locus in relation to the ends of the overlap. Pairs of ts markers recombined to form ts+ progeny in proportion to their distance apart, provided the markers were oriented so that a single crossover between them would produce a full-length genome bearing both ts+ alleles. In the opposite orientation, where such a single crossover would be expected to produce ts/ts recombinants, the ts+ frequency was much lower, indicating that multiple recombination events are rare in this system. These findings rule out site specific recombination, recombination occurring exclusively at the cleaved ends of the overlap, and recombination by means of mismatch repair of a heteroduplex the length of the overlap. They also indicate either that any heteroduplex junction region formed in the course of this reaction is quite short or that it is not subject to heteroduplex repair. Finally, our results demonstrate the efficacy of overlap recombination as a genetic and physical mapping tool and as a method of strain construction, and they suggest other applications, such as using overlap recombination to demonstrate that closely spaced pairs of markers (e.g., putative second-site reversions and their accompanying ts lesions) can be segregated. PMID- 6299006 TI - Identification of the rotaviral gene that codes for hemagglutination and protease enhanced plaque formation. AB - Temperature-sensitive mutants of bovine rotavirus, UK Compton strain, and rhesus monkey rotavirus, MMU18006 strain, were used to derive 16 reassortants by coinfection of MA104 cells. The parental viruses differed phenotypically in their neutralization specificity, their ability to hemagglutinate, and their requirement for exogenous trypsin for infectivity. When the reassortants were assayed for neutralization specificity and hemagglutination, four phenotypes were observed, indicating that these two rotaviral functions segregated independently. Protease-enhanced infectivity phenotype segregated with the HA phenotype indicating that these two functions were manifestations of the same gene product. In order to determine the gene responsible for these rotaviral functions, the reassortants were genotyped by hybridizing 32P-labeled parental transcripts and denatured reassortant genomic RNAs and analyzing the resulting hybrids by gel electrophoresis. The fourth RNA segment was clearly shown to code for HA and protease enhanced plaque formation in MA104 cells. The neutralization antigen was linked to the eighth and ninth RNA segments that comigrated during gel electrophoresis and thus could not be differentiated. PMID- 6299008 TI - Human interferon-alpha-and-gamma-mediated inhibition of retrovirus production in the absence of an inhibitory effect on vesicular stomatitis virus and encephalomyocarditis virus replication in RD-114 cells. AB - RD-114 line is a human sarcoma cell line chronically infected with RD-114 retrovirus. Treatment of these cells with increasing doses of human interferon alpha or -gamma resulted in increasing inhibition of RD-114 virus production. Surprisingly, the replication of vesicular stomatitis virus and encephalomyocarditis virus, in these cells and in the parental RD cells which are not infected with the retrovirus, was insensitive to interferon treatment. Unlike reported differences in other properties of interferon-alpha and interferon gamma, there were no differences in their antiretroviral properties such as dose response, kinetics of establishment of the antiretroviral state, and kinetics of its dissipation upon removal of interferon. PMID- 6299007 TI - Sequential synthesis of small capped RNA transcripts in vitro by vesicular stomatitis virus. AB - Using purified viral or intracellular transcriptive complexes (RNP cores) of vesicular stomatitis virus (VSV), we have identified several small RNA species, ranging in size from 12 to 47 nucleotides in length that are synthesized in vitro by the genomic RNA. One group of small RNA transcripts is composed of three species that are capped at their 5' termini. Two of the capped species are from the start of the N gene and one is from the start of the NS gene. Unlike the previously described 5' triphosphated small RNAs, the templates encoding these small capped RNAs had uv target sizes greater than their respective lengths. In addition, these RNAs appeared sequentially during synchronized in vitro transcription reactions. Thus, these results provide evidence that sequences representing the 5'-capped termini of N and NS mRNAs are synthesized concomitantly with their respective mRNAs rather than simultaneously at the onset of transcription as proposed for the multiple entry, start-stop model (D. Testa, P. K. Chanda, and A. K. Banerjee, 1980, Cell 21, pp. 267-275). Together with the inability of the internally initiated 5'-triphosphated RNAs to be chased into mRNA (R. A. Lazzarini, I. Chien, F. Yang, and J. D. Keene, 1982, J. Gen. Virol. 58, 429-441), these results support a single entry model of VSV mRNA transcription. PMID- 6299009 TI - Characterization of Herpesvirus saimiri and Herpesvirus ateles structural proteins. AB - The structural proteins of Herpesvirus saimiri strains 11 and 11 att and of Herpesvirus ateles strains 73 and 810 were characterized by electrophoresis in SDS-polyacrylamide gels. For H. saimiri 21 virus structural proteins could be identified with molecular weights ranging from 28,000 to 210,000 Da. For H. ateles 810 and H. ateles 73, 20 polypeptides were characterized. Using lactoperoxidase for iodination of surface proteins and immunoprecipitation, 5 polypeptides could be identified as envelope and 4 as capsid surface proteins. PMID- 6299010 TI - Prospective study of posttransfusion hepatitis in cardiac surgery patients receiving only blood or also blood products. AB - The incidence, etiology and risk factors of posttransfusion (PT) hepatitis were evaluated in a prospective study of 297 consecutive open-heart surgery patients. PT hepatitis occurred in 63 (21.2%) patients with a significantly higher hepatitis attack rate in 51 recipients of commercial clotting factor concentrates (56.8%) compared to 246 recipients of blood units from single volunteer donors (13.8% p less than 0.001). Among the concentrates, Prothrombin-complex showed the highest relative hepatitis risk (24) while in patients receiving only blood, the incidence PT hepatitis was correlated with the blood volume transfused. Of the 63 patients with PT hepatitis, 2 (3%) had hepatitis B, 8 (13%) showed evidence of cytomegalovirus infection and 53 (84%) had non-A, non-B hepatitis. These results show that in Italy, as elsewhere, non-A, non-B PT hepatitis is frequent, particularly when commercial blood products are used. PMID- 6299011 TI - Impact of cytomegalovirus testing on blood collection facilities. AB - 561 consecutive O-negative blood donors were tested for the presence of cytomegalovirus (CMV) antibodies using an indirect fluorescent antibody test. 427 (76.1%) donors were CMV antibody positive, while 134 (23.9%) were seronegative. Males (75.1%) and females (78%) did not differ significantly in seropositivity. 17- to 20-year-old males had the lowest frequency of seropositivity (38.5%), though donors in this category represented only 4% of the 561 consecutive donors. The incidence of seropositivity increased consistently with age. Because of the difficulty in confidently generating sufficient CMV-seronegative donors, it is suggested that the exclusive use of frozen-thawed, or washed, leukocyte-poor blood, be evaluated as an alternative. PMID- 6299012 TI - [Tumors of the gonads in male pseudohermaphroditism]. PMID- 6299014 TI - [Mechanism of action of non-steroidal antirheumatic agents]. AB - Non-steroidal antirheumatic drugs permit a nonspecific, symptomatic therapy of inflammatory rheumatic processes and may also be given for a limited period of time in patients with so-called activated osteoarthrosis. Their mode of action is a complex one. More recent knowledge on mediators in inflammation such as prostaglandins, leukotriens and oxygen radicals have brought new insights on the mode of action of non-steroidal antirheumatic drugs. PMID- 6299013 TI - Occupational peripheral neuropathies. AB - Neither clinical nor laboratory evaluation can distinguish occupational neuropathies from neuropathies due to other causes. A worker may suffer either from mechanical injury of individual nerves or from a toxic polyneuropathy that is usually axonal in type. A thorough occupational and environmental history and the recognition of clusters of cases are important in determining the diagnosis. Electrophysiologic studies are helpful in detecting neuropathies in patients who have been occupationally exposed to neurotoxins but have no symptoms. Prevention of occupational neuropathies depends on clinical vigilance, industrial hygiene surveys, biologic monitoring and periodic examination of workers exposed to neurotoxic chemicals. The development of more sophisticated methods of prevention and early detection of peripheral nerve involvement depend on understanding the mechanisms of action of toxins and the pathophysiology of the lesions they cause. PMID- 6299015 TI - Immobilization of glucose isomerase. AB - The immobilization of glucose isomerase (D-xylose ketol isomerase, EC 5.3.1.5) by covalently bonding to various carriers and by adsorption to ion exchange resins was attempted in order to obtain a stable immobilized enzyme which can be used for continuous isomerization of glucose in a column. Of the covalent bonding methods, the colloidal silica-glutaraldehyde method showed the highest binding capacity and gave the most stable immobilized glucose isomerase. The Ludox HS-30 bound glucose isomerase column showed a half-life of 24 days and an enzyme usage of 0.07 units per gram of isomerized sugar (d.s, fructose 45%). Of the resins used, the macromolecular type or porous type strongly basic anion exchange resins showed the highest binding capacity and gave the most stable immobilized glucose isomerase. The Amberlite IRA-904 resine-bound glucose isomerase showed a half life of 23 days and an enzyme usage of 0.06 units per gram of isomerized sugar (d.s., fructose 45%). Based on the ease of the immobilization process, the possibility of carrier reuse and the extensive use already achieved by ion exchange resins in the sugar industry, IRA-904 resin was selected as the candidate for commercialization. PMID- 6299016 TI - Growth characteristics and ultrastructure of protoplast type L-forms from streptomycetes. AB - L-form colonies from S. hygroscopicus, S. griseus and S. levoris were isolated after incubation of lysozyme protoplasts on an osmotically stabilized complex agar medium. Unstable and stable L-forms grow on solid and in liquid media. L form colonies are 5-10 times smaller than normal colonies and show a typical morphology for each species. In ultrathin sections L-form cells are characterized by nucleoid areas with typical core-like structures, by a ribosome-rich cytoplasm with different inclusion bodies, and by a cytoplasmic membrane. Because there are no cell wall structures L-forms of the three Streptomyces species belong to the protoplast type. Analysis of cell size and cell shape shows a variation in diameter and a cell propagation by regular and irregular division- and budding like processes. Many L-form cells contain more than two chromosomes. The results are discussed with regard to the cellular organisation of streptomycetes and the nature of the stable L-form. PMID- 6299017 TI - [Electron microscopic study of the in vitro effect of dipyridamol on the pseudorabies virus]. AB - Dipyridamole at a concentration of 50 microM/ml displays no activity on adsorption and penetration of pseudorabies virus in chicken embryonal cells. Furthermore, first stages of virus replication take place within the nucleus, whereas incomplete virus cores defective in DNA content are found within the nucleoplasm at times when the regular viral replication has been finished in controls. Defective pseudorabies virus particles lacking in DNA-content of the core, can be observed at the end of normal replication time. Consequently, the antiviral activity of dipyridamole may be due to blocking of the synthesis or of the incorporation of infectious viral DNA into the virus core. PMID- 6299018 TI - [Recent findings in biochemistry and the significance of endorphins and enkephalins]. AB - Peptides with morphine-like efficacy, which are called endorphines and enkephalines, are formed by the nerve cells of numerous segments of the central nervous system and of the gastrointestinal canal. The efficacy of these peptides which decrease the sensation of pain is increased in certain stress (lesions, operative interventions, heat, birth). They stimulate the activity of the sympathetic nervous system, influence the secretion of certain hormones and decrease the sensitivity to pain. In acupuncture the release of the endorphines is increased. The formation of the endorphines takes place by peptidolysis from a prodromal molecule--the pro-opiomelanocortin--and the formation of the enkephalines by peptidolysis from the proenkephalines. The binding of the endorphines and of the enkephalines takes place on receptors in the area of the synapses of the certain nerve cells, which also bind opium alkaloids. Under the influence of the opiate-receptor-complexes an influence on the stimulus conduction takes place which is reduced. Particularly those nerve cells are concerned which conduct stimulations from nozireceptors. The endogenic opiates take furthermore part in the regulation of reproduction, the body temperature, the respiration and the function of the gastrointestinal canal. PMID- 6299019 TI - [Laparoscopic aspect and clinical relevance of cholangiofibromas]. AB - In a current series of 1160 laparoscopies in 38 cases (3.2%) we found cholangiofibromas. The picture of single, sporadic and multiple cholangiofibromas as well as cholangiofibromatosis is both described and documented by laparoscopic photography. Clinically it seems remarkable that only patients with alcohol- or drug-induced liver injury showed such alterations. The prevalence of cholangiofibromas in toxic liver damage suggests that we are here not dealing with embryonal aberrations (hamartomas) but apparently with cellular inductive phenomena. PMID- 6299020 TI - [The association of ulcer and cancer is no rarity]. AB - Out of 100 patients with early gastric carcinoma 4 cases with a combined benign gastric and/or duodenal ulcer were encountered. Even in long-lasting peptic ulcer history and multiple benign biopsies carcinoma may develop at some site in the ulcer stomach. That's why intermittent therapy based on ulcer symptoms in a patient with chronic ulcer disease remains problematic. Carcinoma incidence seems to be identical in patients with gastric and duodenal ulcer and does not differ from a population with an ulcer-free stomach. PMID- 6299021 TI - [Diffuse Fabry's angiokeratoma]. PMID- 6299022 TI - [Interaction between neurons of the structures for alimentary and defensive reflexes in Helix pomatia]. PMID- 6299023 TI - [Electronoptical, morphometric and histochemical studies on skeletal muscles of patients with polymyalgia rheumatica]. AB - 1. Skeletal muscle biopsies from 21 patients with polymyalgia rheumatica (PMR) were examined morphometrically, histochemically, and by electron microscopy. 2. Ultrastructural changes in the PMR specimens have been classified according to 15 criteria including nucleus, myofilaments, mitochondria, T-system, glycogen deposition, lipid, lipofuscin and myelin figures. 3. The described focal muscle changes are of regressive character and nonspecific in themselves. 4. The described recording of these criteria shows an extraordinarily high incidence of skeletal muscle changes in PMR. This constellation of characteristics gives a distinctive profile to the ultrastructural picture of the skeletal musculature in PMR. 5. The most important ultrastructural changes concern the mitochondria. Crystalline inclusions and deformations lead to structural changes of the mitochondria which threaten the breathing of the cells. This, however, is compensated by the reproduction and accumulation of the mitochondria. 6. A similar analysis of the regressive muscle changes and the processes in the field of muscular artery media shows morphological analogies pointing to an identical systemic process in PMR and arteritis gigantocellularis. 7. Ultrastructural changes of this kind cannot be explained by inflammatory or noninflammatory arterial occlusions. The same applies to the light microscopic muscle findings. 8. Minor type II fiber atrophy and some moth-eaten and whorled fibers were demonstrated histochemically. These findings are not specific to PMR, being found in skeletal muscle diseases of varying origin. PMID- 6299024 TI - [The functional after-care of the knee joint after plastic surgery of the ligaments in general anterior instability]. PMID- 6299025 TI - [Biology and surgery of small cell bronchial cancer]. AB - The small cell type is encountered in 10 per cent of all operations for lung cancer. It is centrally localised in about 70 per cent, thus pneumonectomy has to be performed as long as the tumour is operable. If the operation is performed as long as lymph node metastases are absent the 5 year survival rate comes up to 39 per cent. This result is in favour of surgery. The 5 year survival rate of all resected cases of small cell carcinomas amounts to about 18 per cent. Paraneoplastic symptoms are rare, they could only be traced in a percentage of 4.5 (n = 389). PMID- 6299026 TI - [Cystosarcoma phyllodes]. PMID- 6299027 TI - [Antibacterial effects of niridazole. II. Effects on aerobic and anaerobic bacteria]. AB - Niridazole which is chemically related to metronidazole is endowed with much better antibacterial activity. First, several genera of aerobic bacteria, such as Salmonella and Escherichia, are susceptible to niridazole, whereas metronidazole is completely ineffective. There exist, however, some particular strains which are naturally resistant to niridazole. The in vitro activity of niridazole is still increased, if these facultative anaerobic bacteria are tested under anaerobic growth conditions. Second, niridazole has a higher in vitro activity against anaerobic bacteria, such as Bacteroides spp., Clostridium spp., Fusobacterium spp. and Peptococcus sp., than metronidazole. Extremely low MIC values of niridazole were found ranging from 0.0037 to 0.06 microgram/ml. Propionibacterium acnes, which are resistant to the action of metronidazole, are also relatively resistant to niridazole. PMID- 6299028 TI - [Pox in the cowled siskin (Carduelis cucullata)]. PMID- 6299029 TI - Clinical bovine virus diarrhoea in Morocco. First report. PMID- 6299030 TI - Immunity of pigs vaccinated by live or inactivated Aujeszky vaccines against experimental infection. PMID- 6299031 TI - [Demyelinization process in acute and subacute experimental encephalomyelitis]. AB - The authors present the results of studying encephalomyelitis caused by the neurotropic (IHM) strain of the murine hepatitis virus in 60 mice of the C3H line infected intracerebrally at the age of 4 weeks. Morphological examinations of the brain carried out on the 5th-13th day (the acute period) and the 14th-30th day (the subacute stage) have shown that it is myelin-producing cells that are affected first in this form of encephalomyelitis, while the periaxonal process observed is a consequence of this affection. Proofs of this conclusion are presented. It is shown that in subacute encephalomyelitis, similar processes develop. Degeneration and infection of oligodendrocytes are not so pronounced in that case, as in the acute disease. However, destruction of even individual oligodendrocytes may lead (due to the peculiarities of their structure and function) to an avalanche-like process of demyelinization. It is supposed that the vesicular degeneration of myelin may arise in the sites of close contact between the myelin membranes and the cells of inflammation infiltrates (release of lysosomal enzymes and toxic products formed in edema). In the latter case the viruses serve as an antigen depot that causes and maintains the inflammatory process. PMID- 6299032 TI - [Neurologic and electroencephalographic evaluations after a single irradiation of pituitary adenomas with protons having 1000 MeV of energy]. AB - The brain was examined (on the basis of neurological and electroencephalographic findings) in 43 patients with hypophysis adenomas both before and at various intervals after a single exposure of the tumour to protons with an energy of 1,000 MeV. The irradiation was performed by the stereotaxic convergent method. The radiation dose absorbed by the tumour was 100 Gy. The local character of the proton action, absence of irreversible changes in the brain, temporary, transient changes on the part of the oculomotor nerves (25.5%) and temporal lobes (9.3%) are demonstrated. The objective remission of the disease was attained in 97.7% of the cases. PMID- 6299033 TI - [Neurologic manifestations of chronic arterial insufficiency of the lower extremities]. AB - Neurological manifestations of lower limb chronic arterial insufficiency were studied in 1004 patients with abdominal aorta and its branches occlusion. Several often met neurological syndromes--neuritic, sympathicoalgetic, radicular (mono-, bi- and polyradicular), plexitis lumbosacralis and spinal, induced mainly by regional hemodynamic disturbances were singled out. Clinical peculiarities of these syndromes helping in differentiation of vascular surgical diseases from neurological ones are shown. PMID- 6299034 TI - [Results of surgical treatment of patients with pituitary adenomas]. AB - The results of operations performed on 126 patients for adenoma of the hypophysis from 1954 to 1975 by the transfrontal approach are analysed. In most patients the adenoma was marked by suprasellar growth. The clinical symptoms and the specific features of the methods of examination and surgical treatment are described. Postoperative mortality was 18.2%, duration of catamnestic follow-up 5-10 years. Recurrences were found in 14% of cases. They occurred 5-8 years, on the average, after the first operation and 1.3 years after repeated operations. The necessity of early diagnosis of hypophyseal adenoma is emphasized. Modified operative techniques and substitution hormonal therapy in combination with timely diagnosis are decisive in improving the results of treatment of patients with adenoma of the hypophysis. PMID- 6299035 TI - Interrelationship between oxidative energy transformation and energy consumption at mitochondrial and cellular levels. AB - The adaptation of oxidative energy transformation in mitochondria to the energy demand of cellular metabolism was investigated in experiments with isolated mitochondria and liver cells and by computer simulation in terms of a mathematical model. Separate draining of different energy pools allowed the determination of the relation between these pools and the elucidation of the importance of the connecting enzyme reactions to the regulation of the whole process. The following conclusions can be drawn from the results: 1. The intramitochondrial adenine nucleotide pool exhibits a homogeneous behaviour, and its changes are the signal for ATP synthesis. 2. The proton-motive force which is in near-equilibrium with the intramitochondrial phosphorylation potential is the immediate signal for the respiratory chain. 3. The intramitochondrial phosphorylation potential is transformed into the external one by a flux dependent non-equilibrium reaction of the translocator. 4. The rate of respiration-linked ATP formation is regulated by more than one reaction step with varying control strength. 5. In both isolated mitochondria and hepatocytes an activation of respiration is provoked by a decrease in the mitochondrial energy state caused by cellular energy utilization. PMID- 6299037 TI - [Detection of IgG and IgM in plasma membrane extracts of lymph node tumors in cattle with enzootic leukosis]. PMID- 6299036 TI - Hereditary spherocytosis: ionophore treatment of erythrocytes in vitro. PMID- 6299038 TI - Basal and TRH stimulated serum levels of TSH in patients with hyperprolactinaemia and in subjects on oestrogen treatment. PMID- 6299039 TI - Active and inactive renin in primary and secondary adrenal insufficiency and during ACTH infusion. PMID- 6299040 TI - Plasma 4-pregnene-17 alpha, 20 alpha-diol-3-one (17 alpha, 20 alpha dihydroxyprogesterone) and 17 alpha-hydroxyprogesterone in man. PMID- 6299041 TI - Effect of a non-steroidal antiandrogen, flutamide, on the hypothalamo-pituitary axis, genital tract and testis in growing male rats: endocrinological and histological data. PMID- 6299042 TI - Multiple species of FSH are present within hamster anterior pituitary cells cultured in vitro. AB - Anterior pituitary cells were obtained from ovariectomized hamsters and cultured for 4 days. The cells were lysed and intracellular proteins, including FSH, were separated by polyacrylamide gel isoelectric focusing. FSH present within cell lysates so separated was quantitated by a specific FSH radioimmunoassay (RIA). Six distinct species of intracellular FSH were observed with pI values = 5.9, 5.7, 5.3, 5.1, 4.7 and 4.3 -- 3.8. All six species were detectable when proteins from a concentrate of incubation medium collected from those cell cultures were separated by isoelectric focusing. Similar isoelectric points were obtained when the FSH species present within an anterior pituitary homogenate obtained from ovariectomized hamsters were separated by the same technique. All but two of these six species of intracellular FSH were well recognized by a hamster FSH receptor preparation. However, these two species of FSH (pI values = 4.7 and 4.3 - 3.8) do not appear to be beta subunits of the intact glycoprotein as determined by gel filtration. Thus, the present studies demonstrate that, as observed previously in anterior pituitary glands obtained from intact and castrated animals, pituitary cells that have been enzymatically dissociated and cultured in vitro for 4 days contain and secrete multiple species of FSH that are separable by polyacrylamide gel isoelectric focusing. This in vitro system may be useful for studying the hormonal factors that regulate FSH species synthesis and secretion. PMID- 6299043 TI - TSH unresponsiveness, a case report. AB - A patient with congenital primary hypothyroidism is presented. His thyroid gland had a normal uptake of radioiodine which was independent of endogenous or exogenous TSH, sympathetic B-receptor blockade or prostaglandin inhibition. Infusion of dibuturyl-cyclic AMP increased the uptake of radioiodine and stimulated release of protein bound 131I. He had no goitre even when he did not receive thyroxine, but thyroid histology showed evidence of active epithelium in the presence of adequate substitution with thyroxine. We assume that some unknown factor other than TSH stimulates part of the glandular function in this patient, without leading to adequate formation and release of thyroid hormone. PMID- 6299044 TI - The renin-angiotensin system in association with hyperreninaemic hypoaldosteronism in neoplasia induced hypercalcaemia. AB - The renin-angiotensin system was examined in Fischer rats at 7, 11 and 14 days after Leydig cell tumour transplantation, and in age matched controls. Mean arterial blood pressure (MAP), active plasma renin and serum calcium were higher (P less than 0.01) in the tumour transplant rats than in the controls at 11 days after transplantation. There was a positive correlation of both active renin and MAP with serum calcium at this time. Although inactive renin levels were elevated in the tumour transplanted rats, the ratio of inactive to active renin was decreased in comparison to controls. Plasma norepinephrine, active renin and plasma angiotensin II were higher in tumour rats at 14 days. Nevertheless, basal levels of aldosterone and MAP as well as aldosterone responses to graded infusion of angiotensin II, ACTH and KCl were decreased in the tumour rats at 14 days. Moderate hypercalcaemia (day 7 and 11), induced by Leydig cell transplantation in the Fischer rat, is associated, therefore with elevated blood pressure which appears to be related, in part, to activation of the renin-angiotensin system. However, severe hypercalcaemia (day 14) was associated with hypotensive hyperreninaemic hypoaldosteronism state. PMID- 6299046 TI - [Acute inhibition of angiotensin-converting enzyme by captopril. Hemodynamic and biochemical aspects in normotension and essential hypertension]. PMID- 6299045 TI - Vitamin E prolongs survival and function of porcine Leydig cells in culture. AB - Vitamin E (alpha-tocopherol) is known to be required for testicular function but its action on specific testicular cells has not yet been studied. The present study used porcine Leydig cell cultures, in a hormone-supplemented medium, to study the effect of vitamin E (vit E) on Leydig cells. It was seen that the addition of vit E to the medium led to an increase in cell survival, lengthening the life span of the cultures from 3-4 days to more than a week. The Leydig cells maintained their LH/hCG receptors and responsiveness throughout this period as evidenced by an increase in testosterone (T) and prostaglandin secretion. The hCG stimulated T levels were synergistically increased in the presence of vitamin E, while basal levels of T secretion were not changed. Other secretory products of Leydig cells are prostaglandins E2 and F2 alpha. It was found that the addition of vit E inhibited both the basal prostaglandin levels and the stimulated levels by 90%. Maximal effects on all of these parameters were seen at 10 ng/ml vit E. It is obvious that vit E plays a critical role in maintaining porcine Leydig cells in primary cultures beyond the first 3 days. This vitamin seems to be involved both in steroidogenesis and in prostaglandin production in the Leydig cells. The exact mechanism of the action of vit E these two biosynthetic pathways remains to be determined. PMID- 6299047 TI - Calcium slow channel blockers: physiology and anesthetic interactions. PMID- 6299048 TI - Enkephalins and endorphins: the endogenous opiates. PMID- 6299049 TI - Central and peripheral nervous system effects of metals: a survey. AB - There are several categories of neurologic dysfunction which have been associated with metal intoxication. Many examples of case histories or animal studies where toxicity by metals has produced clinical manifestations and pathophysiologic changes similar to those found in certain primary neurological disorders have been reported. It is easy to make a diagnosis of metal intoxication in a subject whose work history and circumstances of exposure to specific substances is known. Observation of defined safety standards and time weighted averages for a work day in conjunction with environmental monitoring procedures are intended to reduce risk of undue absorption of known potential intoxicants. However, when a patient is seen by a physician and a probable neurological diagnosis is to be made in the absence of a detailed life-work history, the possible relationship to previous or current intoxication may be overlooked. This communication will review several neurological syndromes which have been found in association with metal accumulation. PMID- 6299051 TI - Elevated beta-endorphin immunoreactivity in umbilical cord blood after complicated delivery. PMID- 6299050 TI - Plasma FSH, LH and prolactin levels in postmenopausal women undergoing cyclofenil treatment. AB - Ten postmenopausal women were studied in an attempt to identify the mechanism of action of cyclofenil, a non-steroidal anti-estrogen which has proved to be effective in the climacteric syndrome. A double-blind inter-patient clinical investigation was undertaken, with patients assigned randomly to treatment for 10 days with cyclofenil, 400 mg/day, or else conjugated estrogens, 1.25 mg/day, always given orally at 8 a.m. Serum FSH, LH and PRL levels were determined daily 2 days before, during and for 2 days after treatment. On the first and tenth day of treatment, five blood samples were drawn between 8 a.m. and 4 p.m. to ascertain if there was any drug-induced variation in the hormonal secretory patterns. Furthermore, endometrial biopsies of all patients were taken before and immediately after the 10-day treatment. In 5 patients, endometrial biopsies were repeated after 3-6 months of therapy with cyclofenil. The results indicate that cyclofenil has two opposing actions on the hypothalamic-hypophyseal axis, one estrogen-like, in that it depresses serum FSH levels, and the other antiestrogen like, in that it depresses serum PRL levels. They also show that at both the peripheral and the central level cyclofenil is a drug of first choice for postmenopausal women at risk for endometrial and mammary neoplastic pathology. PMID- 6299052 TI - The comparative ototoxicities of gentamicin, tobramycin and dibekacin in the guinea pig. A functional and morphological cochlear and vestibular study. PMID- 6299053 TI - Pre-school children with impaired hearing in Goteborg 1964-1980. PMID- 6299054 TI - Vitamin-D-induced hypercalcaemia and its effect on serum gastrin, gastrin cells and antral gastrin in parathyroidectomized rats. AB - Nineteen rats were parathyroidectomized by electrocauterization, resulting in pronounced hypocalcaemia. Seven of these rats were then given vitamin D3 in a dose causing hypercalcaemia, while 7 received a dose which normalized serum calcium and 5 received no vitamin D. Ten further animals were sham-operated and were not given any extra vitamin D. The experimental period was 16 weeks. No significant differences in serum gastrin values were found between the different groups. Quantitative studies of the antral gastrin cells showed no difference in the number of gastrin cells per unit volume or unit segment between the different groups of vitamin-D-treated animals and sham-operated animals. However, the number of gastrin cells per unit segment was significantly higher in the hypocalcaemic animals than in the sham-operated animals, as a result of an increase in mucosal thickness. The amount of antral mucosal gastrin did not differ between the different groups. Thus the results of this study indicate that experimentally induced hypercalcaemia in parathyroidectomized rats does not influence the serum gastrin, the number of gastrin cells or antral gastrin. PMID- 6299056 TI - Occurrence of hepatitis B-surface antigen in a consecutive material of 1539 liver biopsies. AB - The frequency and occurrence of hepatitis B-surface antigen (HBsAg) in 1539 consecutive liver biopsies were determined using orcein staining as screening method. In HBsAg seropositive cases sections were in addition stained with immunoperoxidase staining for HBsAg in order to reassure the presence of HBsAg in liver cells. HBsAg was demonstrated in 36 biopsies (2.3%). The highest frequency was found in biopsies with chronic hepatitis (28%). In biopsies with slight non specific changes 10% and in cirrhotic 4.5% contained HBsAg. HBsAg was not detected in biopsies with acute hepatitis. No correlation was found between the number of HBsAg containing cells in the biopsies and the histological/biochemical activity in the liver. It is concluded that orcein staining of ground-glass appearance is a highly specific HBsAg marker. The sensitivity is about 80% in cases with minimal changes, chronic hepatitis and cirrhosis. The method is unsuitable as etiological marker in cases with acute hepatitis. PMID- 6299055 TI - A scanning and transmission electron microscopic study of pulmonary adenocarcinoma with histological correlation. AB - The three-dimensional growth pattern and cell shape and surface of 26 adenocarcinomas of the lung were studied by scanning electron microscopy. The prevailing subtype according to the WHO histological classification, grade of differentiation and secretion of mucosubstances were determined by light microscopy, the tumour cells being further characterized by transmission electron microscopy. The series included 5 acinar, 12 papillary, 1 bronchiolo-alveolar and 8 solid subtypes of adenocarcinoma. Out of these 5 acinar, 9 papillary and 1 bronchiolo-alveolar carcinomas were well or moderately, and 1 solid carcinoma was moderately differentiated. One acinar, 2 papillary and 7 solid carcinomas were poorly differentiated. All tumours secreted mucosubstances, which in the solid carcinomas occurred as intracytoplasmic vacuoles. In acinar adenocarcinomas there were columnar tumour cells surrounded by fibrotic stroma. In the papillary and bronchiolo-alveolar subtypes the tumour cells formed papillary nodules along the alveolar walls while the solid subtype showed no organized structures. The tumour cells were mostly cylindrical or club-shaped in well and moderately differentiated and spherical in less differentiated neoplasms. In well differentiated papillary carcinomas the tumour cells were tightly attached to each other, whereas many moderately and all poorly differentiated carcinomas had broad intercellular spaces. Tumour cells with microvilli occurred more often in the acinar, papillary and bronchiolo-alveolar subtypes, which were mostly well or moderately differentiated, than in the mostly poorly differentiated solid subtype of pulmonary adenocarcinoma, which often had a rough apical surface with microridges. In conclusion the three-dimensional growth pattern corresponded fairly well with the prevailing histological subtype and the surface structure of cancer cells seemed to reflect the grade of differentiation in pulmonary adenocarcinomas. PMID- 6299057 TI - A Danish adult case of EBV-positive Burkitt's lymphoma. AB - A 50-year-old Danish male presenting with swelling in fossa supraclavicularis and slight swelling of axillary and inguinal lymphnodes was diagnosed histologically as having malignant, non-Hodgkin, blast lymphoma. The EBV association was established by demonstration of EBV-DNA-positive tumor cells and the presence of high titers of antibodies of the IgG class to VCA, EA (D) and EBNA. The unique finding in this case was the high titers of IgA antibodies to VCA and to EA (D). This latter serological picture is characteristic of nasopharyngeal carcinoma (NPC) but has not previously been observed in Burkitt's lymphoma. PMID- 6299058 TI - Cell surface glycoprotein patterns of cell lines derived from human small cell carcinoma of the lung. AB - The expression of major cell surface glycoproteins (sgp) of seven established human pulmonary small cell carcinoma (SCC) cell lines and two autologous non neoplastic lymphoblastoid and fibroblastic cell lines were studied by the galactose oxidase tritiated sodiumborohydride labelling technique. The general sgp pattern of SCC cell lines was different from that of the autologous non malignant cell lines and the various other normal and malignant hematopoietic cells, gliomas and melanomas previously studied by the same technique, thus confirming that the sgp pattern seems to represent a molecular "fingerprint" of various human cell types. The SCC cell lines could be subdivided into two major subgroups with respect to expression of characteristic sgps. One group of lines was characterized by having prominent sgp 52000 (52 K) Dalton (D), 50 KD, 40 KD, 34 KD and 10 KD. In the other group the major sgps had apparent molecular weights of 110 KD, 75 KD and 10 KD, respectively. In addition to these common basic "group specific" sgps each SCC line expressed individually distinct sgps. PMID- 6299059 TI - [Presynaptic action of agkistrodotoxin on neuromuscular transmission]. PMID- 6299060 TI - [Effect of ginsenosides on cardiac performance and hemodynamics of dogs]. PMID- 6299061 TI - [Auto- and hetero-antagonistic actions of drugs]. PMID- 6299062 TI - [Effect of 10-hydroxycamptothecin on adenylate cyclase activity in murine hepatoma cells]. PMID- 6299063 TI - Activation of mouse spleen adenylate cyclase by WR-2721 and the GTP synthetic analogue. PMID- 6299064 TI - [The dexamethasone suppression test in affective disorders. A review]. AB - The Dexamethasone Suppression Test (DST) is a new tool to help in the diagnosis of depression. The authors present the clinical methodology for in- and out patients and review the current literature on the DST. Some show a correlation between cortisol non-suppression after dexamethasone and the diagnosis of endogenous or primary depression. The authors illustrate the interest of the DST by providing there personal experience of this test. The influence of clinical state, severity of depression and other variables, such as age, sex, menopause and family history, are reviewed in reference to the neuro-transmitters control of the DST. PMID- 6299065 TI - The overlapping effects of thyrotropin and gonadotropins on chick embryo gonads in vitro. AB - Pieces of 12- and 15-day-old chick embryo testes and ovaries were cultured in vitro in the presence of thyrotropin (TSH), gonadotropins (FSH + LH) and adrenocorticotropin (ACTH) for different periods. All the explants of treated gonads differentiated into typical testes or ovaries according to their genetic sex. The gonads of 12-and 15-day-old chick embryos showed a good response to both thyrotropic and gonadotropic stimulation. On the other hand, they did not respond to adrenocorticotropic stimulation. Fifteen-day-old chick embryo testes were grown in tissue culture in the presence of the said hormones. Gonadotropins and TSH enhanced the growth and migration of testicular cells as compared with the control or ACTH treated group. In addition, they maintained the germ cells on the upper surface of epithelial cells. These results have confirmed our previous results in vivo in that gonadotropins and thyrotropin hormones accelerated the development of 12- or 15-day-old chick embryo gonads. PMID- 6299066 TI - Recommended deproteinizing methods for plasma guanidino compound analysis by liquid chromatography. PMID- 6299068 TI - Biosynthesis and transport of phosphatidylserine in the cell. PMID- 6299067 TI - Phytates in legumes and cereals. PMID- 6299069 TI - Morphological aspects of dietary fibers in the intestine. PMID- 6299070 TI - Na+ K+-ATPase within neurons and glia in the generation of seizures. PMID- 6299071 TI - Cerebral oxygenation during recurrent seizures. PMID- 6299072 TI - Calcium-calmodulin protein phosphorylation in neuronal transmission: a molecular approach to neuronal excitability and anticonvulsant drug action. PMID- 6299074 TI - Role of benzodiazepine receptors in seizures. PMID- 6299073 TI - Relationship between seizures and cyclic nucleotides in the central nervous system. PMID- 6299075 TI - Role of receptors for neurotransmitters in status epilepticus. PMID- 6299076 TI - New perspectives on c-type cytochromes. PMID- 6299077 TI - Parathyroid hormone: chemistry, biosynthesis, and mode of action. PMID- 6299078 TI - Platelet-activating factor (PAF-acether), an activator of neutrophil functions. AB - The effect of totally synthetic PAF-acether (1-O-octadecyl-2-O-acetyl-sn-glyceryl 3-phosphorylcholine), 2-lyso PAF-acether (1-O-octadecyl-sn-glyceryl-3 phosphorylcholine) and lyso-phosphatidylcholine on enzyme release and superoxide production from human polymorphonuclear neutrophils (PMN were studied. PMN (2 X 10(6) ml-1) were incubated at 37 degrees C with various concentrations of phospholipids in the absence of cytochalasin B. At 10(-7) M, PAF-acether induced superoxide production and beta-glucuronidase, acid phosphatase and lysosyme release, but not that of cytoplasmic lactic dehydrogenase. In the same condition 2-lyso PAF-acether and lyso-phosphatidylcholine were ineffective. In the presence of phagocytic stimuli PAF-acether enhanced in the range from 10(-7) M to 10(-10) M the enzyme release and only at 10(-7) M the superoxide production. Thus, the capacity of PAF-acether to stimulate PMN, as well as platelet function, indicates a prominent role for this lipid mediator in inflammatory processes. PMID- 6299079 TI - Hypotensive activity of PAF-acether in rats. PMID- 6299080 TI - Effects of dithiocarb and (+)-catechin against carbon tetrachloride-alcohol induced liver fibrosis. AB - Treatment of male rats with carbon tetrachloride (CCl4, 2 x weekly 0.2 ml/kg p.o.) and a 5% alcohol solution, instead of drinking water, for 4 weeks led to marked increases in serum enzyme activities (GOT, GPT, SDH), hepatic triglyceride and hydroxyproline content. Diethyl dithiocarbamate (dithiocarb, 200 mg/kg p.o.) simultaneously applied with CCl4 totally suppressed the elevation in serum enzyme activities and hepatic hydroxyproline concentration, and partially suppressed that of the triglyceride content. (+)-Catechin (50-300 mg/kg p.o.) simultaneously applied with CCl4 had no influence on the enhanced serum enzymes, but depressed the augmented content of both hepatic triglyceride and hydroxyproline in a dose dependent way. The most effective dose with respect to the reduction of the hydroxyproline concentration was 100 mg/kg (+)-catechin; the highest dose (300 mg/kg), however, enhanced the CCl4-alcohol-induced hydroxyproline augmentation. PMID- 6299081 TI - [Studies of ocular mouse cytomegalovirus infection]. PMID- 6299082 TI - [Theophylline: mechanism of action, pharmacokinetics and metabolism]. PMID- 6299083 TI - Neutrophil chemotactic factors in asthma. PMID- 6299085 TI - Acute respiratory failure following intravenous verapamil in Duchenne's muscular dystrophy. PMID- 6299084 TI - Radionuclide analysis of right and left ventricular response to exercise in patients with atrial and ventricular septal defects. AB - In patients with ventricular or atrial septal defect, the ventricle which is chronically volume overloaded might not appropriately respond to increased demand for an augmentation in output and thereby might limit total cardiac function. In this study we simultaneously measured right and left ventricular response to exercise in 10 normal individuals, 10 patients with ventricular septal defect (VSD), and 10 patients with atrial septal defect (ASD). The normal subjects increased both right and left ventricular ejection fraction, end-diastolic volume, and stroke volume to achieve a higher cardiac output during exercise. Patients with VSD failed to increase right ventricular ejection fraction, but increased right ventricular end-diastolic volume and stroke volume. Left ventricular end-diastolic volume did not increase in these patients but ejection fraction, stroke volume, and forward left ventricular output achieved during exercise were comparable to the response observed in healthy subjects. In the patients with ASD, no rest-to-exercise change occurred in either right ventricular ejection fraction, end-diastolic volume, or stroke volume. In addition, left ventricular end-diastolic volume failed to increase, and despite an increase in ejection fraction, left ventricular stroke volume remained unchanged from rest to exercise. Therefore, cardiac output was augmented only by the heart rate increase in these patients. Right ventricular function appeared to be the major determinant of total cardiac output during exercise in patients with cardiac septal defects and left-to-right shunt. PMID- 6299086 TI - Precordial S-T segment depression in inferior myocardial infarction. Evaluation by quantitative thallium-201 scintigraphy and technetium-99m ventriculography. PMID- 6299087 TI - Sensitivity of technetium-99m-pyrophosphate scintigraphy in diagnosing cardiac amyloidosis. AB - To determine the value of technetium-99m-pyrophosphate myocardial scintigraphy in the diagnosis of amyloid heart disease this procedure was prospectively performed in 20 consecutive patients with biopsy-proven primary amyloidosis. Eleven patients had echocardiographic abnormalities compatible with amyloid cardiomyopathy, 9 of whom had congestive heart failure. Diffuse myocardial pyrophosphate uptake was of equal or greater intensity than that of the ribs in 9 of the 11 patients with echocardiograms suggestive of amyloidosis, but in only 2 of the 9 with normal echocardiograms, despite abnormal electrocardiograms (p less than 0.01). Increased wall thickness measured by M-mode echocardiography correlated with myocardial pyrophosphate uptake (r = 0.68, p less than 0.01). None of 10 control patients with nonamyloid, nonischemic heart disease had a strongly positive myocardial pyrophosphate uptake. Thus, myocardial technetium 99m-pyrophosphate scanning is a sensitive and specific test for the diagnosis of cardiac amyloidosis in patients with congestive heart failure of obscure origin. It does not appear to be of value for the early detection of cardiac involvement in patients with known primary amyloidosis without echocardiographic abnormalities. PMID- 6299088 TI - Absolute left ventricular volume from gated blood pool imaging with use of esophageal transmission measurement. AB - A new method for determining absolute left ventricular (LV) volume from equilibrium gated blood pool images was validated in 36 patients by comparing gated blood pool (GBP) imaging with contrast ventriculography (CV) using both Simpson's rule (SR) and area-length (AL) calculations. The technique is geometry independent and is the first to correct for tissue attenuation with use of an in vivo point source. An orally administered capsule containing 1 to 2 mCi of technetium-99m (Tc-99m) sulfur colloid is used for this purpose. Left ventricular volumes are determined by dividing attenuation and background-corrected count rates obtained from semiautomated LV regions of interest by the count rate per milliliter from a blood sample. The correlation between GBP and CV (SR) was 0.96 (CV [SR] = 0.99 GBP + 1.32 ml; standard error of the estimate [SEE] = 21.2 ml) for diastole and 0.97 (CV [SR] = 0.93 GBP - 0.03 ml; SEE = 11.9 ml) for systole. The correlation between GBP and CV (AL) was 0.92 (CV [AL] = 0.90 GBP + 16.72 ml; SEE = 27.8 ml) for diastole and 0.95 (CV [AL] = 0.87 GBP + 4.56 ml; SEE = 14.4 ml) for systole. The method is noninvasive and can be performed easily as part of routine gated blood pool imaging and analysis. PMID- 6299089 TI - Long-term effects of dietary fiber on glucose tolerance and gastric emptying in noninsulin-dependent diabetic patients. AB - Effects of long-term (2 months) supplementation of diet with 20 g of guar gum and 10 g of wheat bran on metabolic control was studied in 12 obese, poorly controlled noninsulin-dependent diabetic patients. Addition of fiber reduced urinary excretion of glucose from 30.5 +/- 6 to 8.3 +/- 2 g/24 h, (p less than 0.025), fasting plasma glucose concentration from 301 +/- 24 to 184 +/- 15 mg/dl (p less than 0.025), and plasma cholesterol concentration from 277 +/- 24 to 193 +/- 9 mg/dl (p less than 0.025). No significant changes were observed in the patients weight and serum concentrations of triglycerides, high-density lipoproteins, free fatty acids, and insulin. Addition of fiber also delayed gastric emptying of liquids and solids. This effect became statistically significant 60 and 90 min after intake of a test meal for liquids and solids, respectively. We conclude that addition of guar and bran to the diet resulted in long-term improvement of metabolic control in these patients and that delayed gastric emptying may be one of the mechanisms responsible for this beneficial effect. PMID- 6299090 TI - Impaired intestinal absorption of vitamin D3 in azotemic rats. AB - Changes in vitamin D metabolism and their effect on calcium and bone metabolism in uremia have been extensively studied. However, the possible effect of uremia on intestinal absorption of vitamin D has not been investigated. We determined the rate of intestinal absorption of vitamin D3 in uremic and normal rats using a well-defined in vivo perfusion technique under identical experimental conditions. The rate of jejunal absorption of vitamin D3 in uremic animals (5.09 +/- 1.87 pmol/100 cm/h) was significantly less (p less than 0.001) than that found in the control animals (11.5 +/- 1.6 pmol/100 cm/h). While the underlying mechanism(s) of the observed reduction in vitamin D absorption in uremia is not known, its recognition adds another dimension to the previously recognized abnormalities of vitamin D metabolism in uremia. PMID- 6299091 TI - Interstitial 252Cf neutron therapy for glioblastoma multiforme. AB - 252Cf brachytherapy has been combined with whole brain photon beam therapy to 6000 rads in 5-7 weeks. In early phase I studies, all patients selected for study tolerated the procedure and the subsequent photon beam therapy. All showed improvement in performance status and decreased tumor size by CT scan evaluation, but it became clear that these tumors are of large size and bulk, produce marked adjacent brain edema, and require individualized implant therapy as well as high dose external beam irradiation if response is to occur. PMID- 6299092 TI - Platelets and malignancy. Rationale and experimental design for the VA Cooperative Study of RA-233 in the treatment of cancer. AB - Considerable evidence has accumulated in recent years which implicates blood coagulation reactions in the growth and spread of malignancy. In particular, platelets may accumulate on embolic tumor cells and facilitate their adhesion to the endothelium at distant sites perhaps by enhancing blood coagulation reactions. Alternatively, platelets may promote tumor cell proliferation by contributing a growth-promoting factor or through interactions mediated by prostaglandins. Inhibition of tumor growth and spread by platelet-inhibitory drugs has been demonstrated in several experimental tumor systems. Preliminary data suggest that similar effects may be seen in human malignancy. The purpose of this paper is to review relevant literature which provides the rationale for therapeutic trials of platelet-inhibitory drugs in human malignancy and to describe the experimental design for a trial involving one such drug, RA-233, in a recently established VA Cooperative Study. PMID- 6299093 TI - A randomized phase II study of m-AMSA (NSC 249992) and neocarzinostatin (NSC 157365) in non-small cell bronchogenic carcinoma. An Eastern Cooperative Group Study. AB - Eighty-nine patients with advanced non-small cell bronchogenic carcinoma were treated with either m-AMSA 120 mg/m2 intravenously every 3 weeks or neocarzinostatin 2.0 mg/m2 intravenously daily X 5 every 4 weeks. There were no responses in 40 evaluable patients who received m-AMSA and three partial responses (7.5%) in 40 patients who received neocarzinostatin. Two patients receiving m-AMSA had drug-related deaths. For m-AMSA the major toxicities were hematologic, while for neocarzinostatin the major toxicities were hematologic, gastrointestinal, and fever. We conclude that m-AMSA is inactive while neocarzinostatin has minimal activity in non-small cell bronchogenic carcinoma. PMID- 6299094 TI - Phase II trial of aziridinylbenzoquinone (AZQ) in patients with refractory small cell carcinoma of the lung. AB - Sixteen previously treated patients received AZQ in a phase II study to test therapeutic efficacy in refractory small cell lung cancer. The dose and schedule of AZQ was 20 mg/m2 day 1 and 8, with treatments repeated every 28 days. No objective responses were noted among 16 evaluable patients. Myelosuppression was the major toxicity. AZQ does not appear to have antitumor activity in patients with previously treated small cell carcinoma. PMID- 6299095 TI - Phase II trial of (m-AMSA) 4'-9-(acridinylamino)-methanesulfon-m-aniside in primary liver cancer. PMID- 6299096 TI - Role of antibody to S100 protein in diagnostic pathology. AB - Normal tissues and various tumors were examined for S100 protein, using anti-S100 protein antiserum, in an immunoperoxidase reaction. Among normal tissues, in addition to the previously reported presence of S100 protein in some neurons, glial, and Schwann cells of the nervous system, melanocytes and Langerhans cells of the skin, interdigitating reticulum cells of lymph nodes, and chondrocytes, we demonstrated it in myoepithelial cells and ducts of sweat glands, salivary glands, and the breast, serous glands of the lung, fetal neuroblasts, and sustentacular cells of the adrenal medulla. Among neoplasms, S100 protein previously has been reported in neurogenic tumors, melanomas, and neuroblastomas; we have demonstrated it in mixed sweat gland tumors, histiocytosis X, pleomorphic adenomas of the salivary gland, medullary carcinomas of the breast, bronchioloalveolar carcinomas of the lung, sustentacular cells of pheochromocytomas, teratomas of the ovary, and tumors of cartilage (enchondromas, osteochondromas, and chondrosarcomas). With S100 protein producing tumors, a normal progenitor cell was identified, indicating that demonstration of S100 protein in tumors confirmed their origin. PMID- 6299097 TI - Hyaline globules and intracellular lumina in a hepatocellular carcinoma. AB - The authors report ultrastructural findings on the morphogenesis of hyaline globular inclusions (HG) of a moderately well-differentiated hepatocellular carcinoma, with the features of a fibrolamellar variant. Pale blue cytoplasmic inclusions (pale bodies) of the tumor cells were shown to be intracellular lumina lined with numerous microvilli. Our findings suggest that HG were formed apparently by the successive deposition of fine granular materials in the lumina. Small accumulations were present in the intracellular lumina, which were seen on light microscopy as central hyaline cores of pale inclusions. Larger globular deposits, corresponding to HG and presumably mature ones, were lined closely with membranes devoid of microvilli that were probably shed intraluminally. Our observation was discussed with relation to the previous findings on the inclusions. PMID- 6299098 TI - Virus-associated colitis in homosexual men: two case reports. AB - Two homosexual men with colitis are described. In each case there was evidence of viral infection of the colon. One patient developed toxic megacolon that responded to conservative therapy. The other patient had proctosigmoiditis and opportunistic infections that eventually led to his death. An intensive search for viruses should be made in all homosexual men with proctitis and/or colitis. PMID- 6299099 TI - Cardiovascular, humoral, and renal effects of phenoxybenzamine in hypertension. AB - We examined the effects of alpha adrenergic receptor blockade with phenoxybenzamine on various parameters related to blood pressure regulation in 10 normotensive and 12 essential hypertensive subjects. The responses were observed before and after phenoxybenzamine treatment during standardized maneuvers of volume expansion with saline infusion and volume contraction with a diuretic. Alpha adrenergic blockade produced a significantly greater (P less than 0.02) reduction in blood pressure in the hypertensive group than in the normotensive subjects. The baroreceptor response, evaluated by comparing the change in plasma norepinephrine concentration in relation to the change in blood pressure, was significantly reduced (P less than 0.05) in hypertensives compared to normal subjects. Plasma and urinary norepinephrine concentration, which were similar in the two groups, increased in both during phenoxybenzamine treatment. There was no significant change in the mean levels of plasma renin activity or plasma aldosterone concentration in either group after phenoxybenzamine treatment. However, the relationship between plasma renin activity and plasma aldosterone levels, as judged by linear regression analysis, was significantly altered (P less than 0.05) by phenoxybenzamine therapy in both groups. The latter may reflect an effect of phenoxybenzamine on the metabolism or secretion of aldosterone. These results, utilizing an alpha adrenergic antagonist, confirm the enhanced vascular reactivity and diminished baroreceptor function in essential hypertension; no evidence for an alpha adrenergic effect on plasma renin activity could be demonstrated in normotensive or hypertensive subjects. PMID- 6299100 TI - Urinary retention as the presenting symptom of acquired cytomegalovirus infection. PMID- 6299101 TI - Deletion mapping of polymorphic loci by apparent parental exclusion. AB - Deletion of a chromosome region containing a polymorphic marker may result in apparent parental exclusion at that locus. We present a general method for calculating the probability that deletion at a specific locus would have such an effect. For many autosomal loci this probability is substantial, justifying attempts at deletion mapping in most cases. This method may be especially valuable in assigning DNA restriction fragment polymorphisms to chromosome regions. PMID- 6299102 TI - Effect of adrenocorticotropic hormone and human chorionic gonadotropin before and after bilateral oophorectomy in a patient with acquired adult-onset adrenal hyperplasia: a case report. AB - A 29-year-old woman presented with hirsutism, obesity, oligomenorrhea, and infertility caused by oligoovulation and tubal occlusion. Partial 21-hydroxylase deficiency of the adrenal was suggested by an abnormal adrenocorticotropic hormone (ACTH) stimulation test. The patient subsequently developed bilateral tuboovarian abscesses and underwent abdominal hysterectomy and bilateral salpingo oophorectomy. Thus, an opportunity was presented to study the adrenal endocrine disorder in the absence of ovaries and to investigate the effect of human chorionic gonadotropin (hCG) on the adrenals. There was no change in the adrenal response to ACTH stimulation after oophorectomy. hCG stimulation resulted in an increase in dehydroepiandrosterone sulfate and 17 beta-estradiol levels, suggesting that hCG had a stimulatory effect on the adrenal. The ovarian-adrenal relationship and effects of adrenal stimulation in the absence of ovaries are discussed. PMID- 6299103 TI - Oral ketoconazole therapy for keratomycosis. AB - Two patients, a 48-year-old woman and a 44-year-old man, with fungal keratitis were treated with orally administered ketoconazole, a new antifungal agent. In both cases healing and regression of the corneal ulcer began four or five days after initiation of ketoconazole treatment (300 mg/day). The clinical signs of corneal infection disappeared after eight (Case 1) and three (Case 2) weeks and visual acuities improved markedly (from counting fingers at 5 cm to 20/30 in Case 1 and from 20/30 to 20/16 in Case 2). The levels of serum glutamic oxaloacetic transaminase and serum glutamic pyruvic transaminase increased slightly during the course of treatment (total dosages: 13,000 mg in Case 1 and 4,700 mg in Case 2), but no other systemic or topical signs of toxicity or adverse reactions were noted in either case. PMID- 6299104 TI - Papillomavirus in human conjunctival papillomas. AB - Forty-one human conjunctival papillomas were histopathologically examined for koilocytosis and papillomavirus common antigen with a peroxidase-antiperoxidase technique immunospecific for genus-specific papillomavirus antigen. Koilocytotic cells, typically found in other papillomavirus-induced tumors, were present in 24 of the 41 specimens. Two of the 41 specimens were positive for viral antigen. The positive reactions were intranuclear within the superficial layers of the epithelium. PMID- 6299105 TI - Orchestration of tooth movement. AB - Recent advances in the art and mechanics of delivering orthodontic forces have achieved greater precision and control of tooth movement. The exact mechanism by which these forces orchestrate tooth movement is not thoroughly understood. An accurate understanding and precise control over the factors responsible for initiating and carrying out the tissue reaction will ultimately optimize the rate of tooth movement. An integrated hypothetical model for the mechanism of tooth movement is discussed here. This model is based on the most recent body of information available to explain how various stimuli affect bone cells. The roles played by piezoelectric responses, prostaglandin production, and biochemical factors are discussed, with particular emphasis on their importance and contribution in terms of maximizing the rate of tooth movement. The intelligent use of this knowledge will permit us to modify the orthodontic appliance and treatment regimen in order to achieve an optimum tissue response. PMID- 6299107 TI - Electron spin resonance dating of human bones from Brazilian shell-mounds (Sambaquis). AB - Electron spin resonance (ESR) signals from bone increase with exposure to radiation. This permits the dating of ancient bone from its exposure to natural radiation over the centuries. The ESR technique was used for dating human bones from Brazilian shell mounds. The results were compared with 14C dates on charcoal found near the bone. The natural radiation dose rate of the bones was about 0.01 Gy/year (1 rad/year), similar to that found in Japanese shell-mounds. Ages of the bone samples dated ranged from 2000-5000 years BP. PMID- 6299106 TI - Aggregation of equine platelets by Onchocerca cervicalis collagen. AB - The arachidonic acid pathway plays an important role in many inflammatory reactions. Current evidence suggests that platelets can play a central part in host inflammation. Since microfilariae are mobilized into the bloodstream following diethylcarbamazine (DEC) treatment, we have studied the effects of Onchocerca cervicalis cuticle preparations on equine platelet aggregation. The authors have found that O cervicalis cuticular preparations can induce platelet aggregation in vitro. Furthermore, this activity was abrogated by treatment with collagenase and not hyaluronidase, elastase, or alpha-chymotrypsin. When this evidence is viewed collectively with the evidence for in vivo parasite cuticular damage following DEC treatment, it becomes entirely plausible that the cuticular damage may indeed reveal a platelet-reactive surface, thus permitting platelet parasite binding to occur. This binding would result in platelet aggregation and the generation and release of platelet-derived arachidonate metabolites. These metabolites may play a very critical role in the development of the described pathologic sequelae observed following DEC treatment. Field studies using cyclooxygenase and lipoxygenase inhibitors might therefore be very efficacious in decreasing the frequency of side effects due to DEC or other potentially effective drug regimens. PMID- 6299108 TI - Trophic response of rat brown fat by glucose feeding: involvement of sympathetic nervous system. AB - Brown adipose tissue has earlier been suggested as an important site of the diet induced thermogenesis that results from cafeteria feeding in rats. The aim of the present communication has been to see if any defined component of this diet can mimic the effects of the diet on the trophic response of brown fat and if these effects are mediated by the sympathetic nervous system. Rats fed a lipid emulsion did not show hypertrophy of brown adipose tissue. Rats fed a glucose solution, whether voluntarily or by force feeding, showed a clear trophic response of brown fat, as seen by the morphology of the tissue and its increased wet weight, increased protein content, increased total and specific cytochrome c oxidase activity, and increased mitochondrial guanosine diphosphate binding. Chemical sympathectomy of young rats by guanethidine prior to glucose feeding impaired the glucose-induced effects on brown fat. beta-Adrenergic blockade in adult rats also tended to depress the glucose effect. Consequently we conclude that chronic glucose ingestion can mimic cafeteria feeding with respect to the trophic response of brown fat and that an intact sympathetic nervous system is required for the mediation of the glucose effect to the brown adipose tissue. PMID- 6299110 TI - Effect of alloxan diabetes on a Ca2+-activated proteinase in rat skeletal muscle. AB - The effect of making rats diabetic by alloxan injection on activity of the muscle Ca2+-activated proteinase (CAF) was investigated. Groups of four to seven control or alloxan-injected rats were killed 10 min (0 day) and 10, 17, and 24 days after a second alloxan injection. The second alloxan injection was given 3 days after the first. CAF activity was assayed in fractions precipitated between 0 and 45% ammonium sulfate saturation (P0-45 crude CAF fractions) that had been prepared so as to remove the protein inhibitor of CAF. Gel permeation, followed by DEAE cellulose chromatography of pooled P0-45 crude CAF fractions from each time treatment group, demonstrated that the assays used in this study were specific for CAF activity. Muscle CAF activity was up to 50% higher in alloxan-injected rats than in control rats, regardless of whether activity was expressed per gram sarcoplasmic protein, per gram contractile protein, or per gram skeletal muscle fresh weight. Alloxan injection diminished rate of muscle mass accumulation but did not change the proportion of sarcoplasmic or contractile protein in skeletal muscle. Hence, alloxan injection decreased the rate of contractile protein deposition. The elevation of muscle CAF activity in alloxan-injected rats is consistent with the proposed role of CAF in initiating metabolic turnover of myofibrillar proteins but does not prove this role nor exclude participation of other proteinases. PMID- 6299109 TI - Differential response to hormones of defined distal nephron epithelia in culture. AB - Defined cultures of rabbit kidney cortical collecting tubule (CCT) and cortical thick ascending limb of Henle's loop (CAL) were grown in monolayers from individual microdissected tubules and maintained for up to five passages, a maximum of 53 days. CCT cells contained cytochemically demonstrable vasopressin stimulated adenylate cyclase, whereas CAL cells were characterized by the localization of Na+-K+-ATPase. [3H]thymidine labeling index decreased with time in primary cultures in the presence or absence of 3% serum. When added to unsupplemented serum-free media alone or in combinations, the growth factors dexamethasone, thyroxine, insulin, epidermal growth factor, and prolactin stimulated [3H]thymidine incorporation to different extents. CCT cells were maximally stimulated by addition of dexamethasone alone, whereas a combination of dexamethasone, thyroxine, insulin, and prolactin was most stimulatory for CAL cells. Addition of hormones concerned with renal ion and water transport to fully supplemented serum-free media inhibited [3H]thymidine labeling index: 1) vasopressin, isoproterenol, and dibutyryl cAMP were equally inhibitory in CCT and CAL cultures; 2) parathyroid hormone and prostaglandin E1 were more inhibitory in CAL cultures; and 3) aldosterone was particularly inhibitory in CCT cultures. PMID- 6299111 TI - Na+-K+-ATPase in rat vascular smooth muscle cell grown in vitro. AB - This study has focused on the characteristics of the Na+-K+-ATPase in in vitro preparations of vascular smooth muscle cells (VSMCs) derived from the rat carotid artery. The maximum velocity of enzyme reaction (Vmax) for the specific activity of the enzyme in the VSMCs' preparations was 2.36 +/- 0.04 (SE) mumol Pi X mg cell protein-1 X h-1 or 0.82 +/- 0.02 mumol Pi X 10(6) cells-1 X h-1. The activation of the enzyme by potassium, sodium and ATP has been investigated. The half-maximal values for potassium and sodium activation of the enzyme in the preparations were 1.18 and 10-20 meq/l, respectively. The respective Vmax values for potassium and sodium activation were reached at concentrations of 4-10 and 80 100 meq/l. The Michaelis constant for ATP was 0.83 mM. Calcium exerted a potent inhibition on the activity of the enzyme (I50 at 1 mM). It has been concluded that the Na+-K+-ATPase kinetic pattern in in vitro preparations of VSMCs is quite similar to that observed in homogenates or subcellular fractions of other tissues. PMID- 6299113 TI - Glucocorticoids and appearance of 1,25-dihydroxyvitamin D3 receptor in rat intestine. AB - The ontogenesis of the 1,25-dihydroxyvitamin D3 specific binding activity in intestine was examined in vitamin D-deficient and replete rats. The absence of binding activity in intestines during the first two postnatal weeks was not influenced by vitamin D supplementation. The concentration of binding sites peaked on day 18 in vitamin D-replete rats and preceded that in the deficient group by approximately 1 wk. The influence of glucocorticoids on 1,25 dihydroxyvitamin D3-binding protein levels was examined by sequential hydrocortisone administration and adrenalectomy. Subcutaneous hydrocortisone administration before day 14 postpartum did not induce binding activity. The concentration of binding sites was significantly increased to 369 +/- 60 fmol/mg of protein by hydrocortisone injections from days 15 to 17 postpartum when compared with an average of 182 +/- 16 fmol/mg of protein in littermate controls. Hydrocortisone administration did not further increase receptor levels in rats injected from days 19 to 21. Bilateral adrenalectomy on day 17 postpartum significantly decreased the concentration of binding sites. It is concluded that adrenal glucocorticoids play an important role in the developmental appearance of 1,25-dihydroxyvitamin D3 specific binding activity in the postnatal rat intestine. PMID- 6299112 TI - Calcium flow-independent actions of calcium channel blockers in toad urinary bladder. AB - A role for transmembrane calcium movement in vasopressin stimulation of its target cell has been postulated based on studies with calcium entry blockers such as verapamil. We examined the effect of three sets of structurally different calcium blockers--D600 (an analogue of verapamil), diltiazem, and nifedipine--on water flow in toad bladder. D600 (200 microM), diltiazem (200 microM), and nifedipine (60 microM) inhibited vasopressin-induced water flow but enhanced adenosine 3',5'-cyclic monophosphate (cAMP)-induced water flow, suggesting that the drugs inhibit cAMP generation in response to vasopressin but enhance the response to exogenous cAMP by inhibiting phosphodiesterase activity. In the case of vasopressin stimulation, inhibition of cAMP generation appears to be the overriding effect. This was confirmed by measurements of cAMP content and the protein kinase ratio (-cAMP/+cAMP), which were significantly lower in bladders receiving both D600 and vasopressin than in those receiving vasopressin alone. Furthermore the drugs inhibited activation of adenylate cyclase by vasopressin in cell homogenates and inhibited phosphodiesterase in both homogenates and membrane free supernatants. Thus these "calcium channel blockers" can directly alter cAMP metabolism in settings where movement of calcium should be irrelevant. The close correlation between the biochemical and transport effects of these agents suggests that their effect on water flow may occur by a direct effect on cellular enzymes or the membranes in which they reside and not by altering local calcium concentrations. PMID- 6299114 TI - Role of the electrochemical gradient for Na+ in D-glucose transport by mullet kidney. AB - Brush border membrane vesicles were prepared from striped mullet (Mugil cephalus) kidney. As judged by the marker enzyme activities, these membranes were enriched 10- to 15-fold compared with the initial homogenate. Uptake of D-glucose was phlorizin sensitive and Na+ dependent. L-glucose uptake was unaffected by either phlorizin or Na+. D-Glucose uptake reflected entry into the osmotically active intravesicular space, not binding to the membranes. When Na+ was increased from 25 to 100 mM, the maximum velocity of glucose uptake was increased from 92 to 423 pmol . s-1 . mg protein-1, whereas the apparent Km (1.27 +/- 0.23) was not altered as Na+ increased. 22Na+ uptake by these membrane vesicles was stimulated by D-glucose and inhibited by phlorizin. These results indicated that Na+ and glucose entered the vesicles via a cotransport process. Consistent with this interpretation, it was possible to show that glucose uptake could be driven by either the chemical or electrical component of the Na+ electrochemical gradient and that the contributions of these two components were additive when both were present. Finally, it was shown that the coupling ratio between Na+ and glucose was approximately 1:1. Accordingly, these results indicate that reabsorption of D glucose across the brush border membranes is coupled to the transmembrane electrochemical gradient of sodium ions. PMID- 6299115 TI - Canalicular bile salt-independent bile formation: concepts and clues from electrolyte transport in rat liver. AB - Studies on canalicular electrolyte transport are reviewed with reference to the concept that hepatocellular inorganic ion secretion may provide an osmotic drive for canalicular water flow. Cellular transport of electrolytes and of some nonelectrolytes appears directly or indirectly (cotransport or potential sensitive transport) related to the activity of Na+-K+-ATPase of the sinusoidal cell membrane, but the role of the enzyme in regulating bile flow remains undetermined. Bile secretion of the isolated rat liver continues in the absence of either Na+, K+, Cl-, or HCO-3 when these ions are replaced in the perfusion medium by other permanent ions. Transepithelial salt concentration gradients, established experimentally, cause transient changes of bile flow and dissipate very quickly. Isotopic ion equilibration between sinusoids and bile proceeds faster than between sinusoids and liver cells. Both observations indicate extensive electrolyte diffusion through a paracellular shunt pathway. This pathway appears preferentially permeable to cations, and it restricts permeation of molecules of the size of sucrose (no apparent diffusion or effects of solvent drag) or bile acids (no backleak). In promoting canalicular osmotic water flow, transepithelial concentration gradients of NaCl are less effective than those of sucrose, revealing a reflection coefficient of NaCl of 0.3. By perfusion with hypertonic medium containing sucrose, bile flow is reduced. Bile production against this opposing osmotic gradient is accomplished by an increase in biliary organic anion concentration. Inorganic ion concentrations essentially conform to a Gibbs-Donnan distribution across the canalicular epithelium, established by the presence of impermeant anions in bile. Hence, the luminal electrical potential is expected to be negative with respect to the sinusoids. It is concluded that biliary secretion of endogenous organic anions is the major osmotic driving force for canalicular bile salt-independent bile flow and that transport of inorganic ions into bile results mainly from diffusion and solvent drag. PMID- 6299116 TI - Measurement of cholecystokinin octapeptide-induced motility of rat antrum, pylorus, and duodenum in vitro. AB - Motor effects of cholecystokinin octapeptide (CCK-OP) on rat antrum, pylorus, and duodenum have been studied in vitro under standard conditions. Intraluminal pressure changes were simultaneously measured at the three locations using a perfusion manometric system with a novel intraluminal pressure-sensor device. This device comprised an acrylic cast of the rat gastroduodenal tract containing the perfusion catheters that reached the surface of the cast with their outlets in the antrum, pylorus, and duodenum. A selective and sensitive intraluminal local pressure measurement was achieved with this pressure sensor due to its shape. CCK-OP increased base-line pressure in the antrum, pylorus, and duodenum; frequencies of phasic contractions in the antrum and pylorus; and amplitudes in the duodenum. The peptide also decreased contraction amplitudes in the antrum and pylorus and frequency of phasic contractions in the duodenum. It is concluded that the novel intraluminal pressure sensor is a useful tool for measuring local pressure changes in the gastroduodenal tract of the rat. In this experimental model, effects of CCK-OP on antral, pyloric, and duodenal base-line pressure are comparable with those observed in isolated muscle strips and in the intact organ of humans, dogs, and opossums. A different behavior, however, was observed in the force of antral and frequency of duodenal phasic contractions. PMID- 6299117 TI - Mechanism of action of cholecystokinin octapeptide on rat antrum, pylorus, and duodenum. AB - The mechanism of action of cholecystokinin octapeptide (CCK-OP) on tonic and phasic contraction of antral, pyloric, and duodenal smooth muscles was studied with a novel perfusion manometric system in isolated esophagogastroduodenal preparations of the rat. CCK-OP increased baseline pressure at each site, frequencies of phasic contractions in the antrum and pylorus, and amplitudes in the duodenum. It decreased antral and pyloric amplitudes and frequency of duodenal phasic contractions. CCK-OP action on tonic contraction was tetradotoxin (TTX) susceptible and its action on phasic contractions was TTX resistant. Phentolamine, phenoxybenzamine, propranolol, catecholamine depletion of preparations by reserpine-tetrabenazine, and the block of catecholamine synthesis at different levels significantly inhibited CCK-OP-induced tonic contraction, whereas atropine had no influence. Adrenergic and cholinergic neural actions on phasic contractions altered the level of amplitudes and frequencies on which CCK OP action occurred. It is concluded that CCK-OP action on tonic contraction of the rat gastroduodenal junction is mediated by a neural noncholinergic pathway, whereas its effect on muscles responsible for phasic contractions is a direct one. PMID- 6299119 TI - Effect of synthetic diets on gastrointestinal mucosal DNA synthesis in rats. PMID- 6299118 TI - Effect of lanthanum on pancreatic protein synthesis in streptozotocin-diabetic rats. AB - The role of extracellular Ca2+ in mediating the stimulatory effect of cholecystokinin octapeptide (CCK8) on [3H]phenylalanine incorporation into protein was studied in isolated pancreatic acini from streptozotocin-diabetic rats. The stimulatory effect of CCK8 (10(-10) M) on [3H]phenylalanine incorporation was completely abolished by preincubating acini with either 10(-4) M lanthanum or 10(-3) M manganese. At these concentrations neither compound altered the basal rate of amino acid incorporation, and both compounds inhibited CCK-mediated Ca2+ influx without affecting either basal or CCK-mediated 45Ca2+ efflux. Lanthanum (10(-4) M) also blocked the stimulatory effect of the cholinergic analogue carbachol (10(-5) M) on amino acid incorporation but did not alter the stimulatory effect of insulin (1.67 x 10(-8) M). Vasoactive intestinal polypeptide and 12-O-tetradecanoyl-phorbol-13-acetate failed to increase the incorporation of [3H]phenylalanine into acinar protein. These findings suggest that CCK and other pancreatic secretagogues that act via Ca2+ enhance protein synthesis by increasing cell membrane permeability to Ca2+ and provide additional evidence that this may be an important mechanism by which CCK regulates pancreatic exocrine function. PMID- 6299120 TI - Myocardial relaxation. VI. Effects of beta-adrenergic tone and asynchrony on LV relaxation rate. AB - We studied the effect of left ventricular (LV) asynchrony and alterations in beta adrenergic tone on the systolic load (pressure) dependency of LV isovolumic relaxation rate in anesthetized dogs. The time constant (T) of isovolumic exponential pressure decline was used as an index of relaxation rate. Variably afterloaded LV contractions resulted in a progressive increase in LV end-systolic pressure from 124 +/- 6 in the control beat to 176 +/- 11 mmHg in the third beat and a progressive lengthening of T from 19 +/- 2 to 30 +/- 4 ms. The direct relation between LV end-systolic pressure and T was nearly linear (r = 0.98), and the slope (k) of this relation was taken to reflect the systolic load dependency of T. Administration of isoproterenol (n = 6) produced a decrease in k from 0.11 +/- 0.02 to 0.08 +/- 0.02 (P less than 0.05); with propranolol (n = 6), k increased from 0.08 +/- 0.02 to 0.27 +/- 0.04 (P less than 0.01). Right ventricular epicardial pacing (n = 6) produced an asynchronous LV contraction and an increase in k from 0.09 +/- 0.02 (atrial pacing) to 0.25 +/- 0.04 (P less than 0.01). These studies confirm the dependency of LV relaxation rate on systolic loads and indicate that this form of load-dependent relaxation can be modified by alterations in beta-adrenergic tone and LV asynchrony. The observed alterations suggest the importance of temporal dispersion of the contraction-relaxation sequence as a mechanism responsible for disturbed relaxation. PMID- 6299121 TI - Effect of caloric restriction on cardiac reactivity and beta-adrenoceptor concentration. AB - The effect of a 21-day program of caloric restriction on cardiac reactivity and beta-adrenoceptor number was investigated in male Sprague-Dawley rats. Rats on the restricted diet (Restricted) exhibited significant decreases in body weight, epididymal fat pad, and retroperitoneal fat pad weight as well as the percent of body fat represented by these adipose tissue depots when compared with rats fed ad libitum (Fed). Fed rats exhibited significantly increased total heart weight and total heart protein, but the percent cardiac protein and ratio of heart weight to body weight were similar in Fed and Restricted rats. Isolated atria from Fed and Restricted rats developed similar chronotropic and inotropic responses over a range of isoproterenol concentrations. Although total beta adrenoceptor number (fmol/heart) was greater in Fed rats, the concentration of beta-adrenoceptors (fmol/mg protein) was remarkably similar regardless of the dietary regimen. Therefore, despite significant decreases in body weight, body fat, and heart weight, the myocardium of Restricted rats maintained the capability of responding to isoproterenol as that of Fed rats, the mechanism of which is at least partially mediated through maintenance of beta-adrenoceptor concentration. PMID- 6299122 TI - Atlantic hagfish cardiac muscle: metabolic basis of tolerance to anoxia. AB - Oxygen tensions in the major venous inputs to the systemic and portal-vein hearts of normoxic Atlantic hagfish (12.3 +/- 1.7 and 11.0 +/- 1.6 mmHg, respectively) are low compared with typical vertebrate values. Anoxia and poisoning with cyanide and azide do not significantly affect in situ performance of the systemic heart. Idoacetate poisoning, however, results in a significant decrease in cardiac performance of the systemic heart to 12% of the initial value after 3 h. Activities of mitochondrial enzymes of hagfish ventricle suggest a small potential for aerobic metabolism compared with those in the aerobic ventricle of Atlantic cod. Activities of enzymes of carbohydrate metabolism indicate similar anaerobic capacity in hagfish and cod ventricle. The ratio of pyruvate kinase to cytochrome c oxidase, an index of anaerobic to aerobic capacity, is 5.6 times greater in hagfish than cod ventricle. Metabolite concentrations in freeze clamped ventricles of normoxic and hypoxic hagfish indicate hypoxia-induced activation of glycogenolysis, enhanced substrate flow across 6 phosphofructokinase, and an apparent secondary constriction of glycolysis at the level of glyceraldehyde-phosphate dehydrogenase. Carbohydrate utilization via the glycolytic pathway appears essential for maintenance of cardiac performance in both normoxic and anoxic hagfish. Under conditions of severe hypoxia, ATP provision is probably met by anaerobic glycolysis. PMID- 6299123 TI - Paraventricular nucleus region controls pituitary-adrenal function in Brattleboro rats. AB - To study the role of the paraventricular nucleus and of neurohypophysial hormones in the control of ACTH secretion, the paraventricular nuclei (PV) of Brattleboro diabetes insipidus rats (DI) were lesioned (L) with a knife; sham-lesioned DI (S) served as controls. Four days later, the rats were stressed by ether inhalation, and blood samples were taken during and 30-40 min after stress for the determination of corticosterone. The median eminence (ME) and neural lobe (NL) were homogenized in 50 microliters of 0.1 N HCl and frozen pending bioassay of corticotropin-releasing factor (CRF). PV lesion almost abolished the corticosterone secretion to ether and reduced the ME CRF content three- to sevenfold. The NL CRF content in S was about twice that of ME, and oxytocin accounted for more than 60% of NL CRF. However, PV lesion had no effect on NL CRF activity. Low amounts of oxytocin (2 mU/ml) had no significant CRF activity but potentiated the ME CRF effect in L. The results suggest that 1) PV is one of the most important sites for CRF synthesis or CRF fiber transit in DI; 2) corticosterone secretion to ether stress is governed mainly by ME CRF; and 3) a large proportion of CRF fibers to NL probably originates outside PV. PMID- 6299124 TI - Papillomavirus infection of the cervix. II. Relationship to intraepithelial neoplasia based on the presence of specific viral structural proteins. AB - Three hundred twenty-two cases of cervical dysplasia (mild, moderate, and severe) and carcinoma in situ (CIS) were examined for the presence of papillomavirus structural antigens with a peroxidase-antiperoxidase method on formalin-fixed, paraffin-embedded tissue. The primary antiserum, prepared from purified, detergent-disrupted bovine papillomavirus type 1 virions, is broadly reactive against the genus-specific (common) antigen(s) of the papillomaviruses. Using the peroxidase-antiperoxidase technique on cervical tissue obtained from biopsy, conization and hysterectomy specimens, papillomavirus structural proteins were identified in association with mild dysplasia in 65 of 152 (43%) cases, with moderate dysplasia in 12 of 82 (15%) cases, with severe dysplasia in eight of 47 (17%) cases, and with CIS in four of 41 (10%) cases. Papillomavirus antigens were found directly within the lesion in all the cases of mild and moderate dysplasia but in only two instances of severe dysplasia and in none of the examples of CIS. In the remaining 10 cases of severe dysplasia and CIS associated with the presence of papillomavirus antigens, cells containing papillomavirus structural proteins were present in areas of moderate dysplasia immediately adjacent to the high-grade lesions in seven instances and in areas of mild or moderate dysplasia not directly in contact with the high-grade lesions in three. Among the 12 high grade lesions associated with the presence of papillomavirus antigens, a morphologic transition from areas of moderate dysplasia containing papillomavirus antigens to the areas of severe dysplasia and CIS was present in five instances. The results of this study, therefore, provide direct evidence demonstrating the relationship of papillomavirus to intraepithelial cervical neoplasia ranging from mild dysplasia to severe dysplasia and CIS. PMID- 6299125 TI - A combined germ cell--gonadal stromal--epithelial tumor of the ovary. AB - This report describes a previously unreported type of ovarian tumor containing three distinct components: germ cells, gonadal stromal cells, and epithelial cells. The germ cells were mainly of the types seen in various stages of oogenesis. The gonadal stromal component had two lines of differentiation. Cells containing Charcot-Bottcher filaments were identified as Sertoli cells and granulosa cells were characterized by a microfollicular arrangement. The epithelial component resembled the cells present on the ovarian surface and lacked specific differentiation toward any of the common ovarian (mullerian) epithelial cell lines. The tumor was discovered as an abdominal mass in a newborn girl with a 46XX karyotype. Subsequently, the child has developed normally and is well 7 years after surgery. PMID- 6299126 TI - [Selective stimulation of utero-placental circulation for the purpose of treating experimental fetal hypoxia]. PMID- 6299127 TI - On the nature and nomenclature of a primary small carcinoma of the skin exhibiting endocrine (? Merkel cell) differentiation. PMID- 6299128 TI - Unusual cutaneous carcinoma with features of small cell (oat cell-like) and squamous cell carcinomas. A variant of malignant Merkel cell neoplasm. AB - Three unusual primary neoplasms in the skin that occurred in elderly patients (64, 77, and 69 years of age) are presented. Two histologic components were found in each of the three neoplasms, one being small cell or oat cell-like carcinoma and the other squamous cell carcinoma. Study by electron microscopy revealed neurosecretory granules, though few, in the small cell component. The lack of clinical evidence of oat cell carcinoma elsewhere, the autopsy finding of two histologic components in one patient, and the findings by conventional and electron microscopy lead us to conclude that these three neoplasms are primary in the skin and of Merkel cell origin. We suggest further that small cell carcinoma of the skin represents a poorly differentiated counterpart of trabecular carcinoma. The spectrum of malignant Merkel cell neoplasm is, therefore, expanding. Two of our three patients died of their neoplasms 2 and 2 1/2 years, respectively, after the initial diagnosis. PMID- 6299130 TI - Column chromatography on polyethylenimine-silica: rapid resolution of nucleotides and proteins with short columns and low pressures. PMID- 6299129 TI - [Neuromuscular and cardiovascular effects of Duador, a new short-acting nondepolarizing muscle relaxant]. AB - The neuromuscular blocking and circulatory effects of a recently synthesized relatively short acting nondepolarizing muscle relaxant, Duador, have been investigated in anaesthetized patients. Duador appears to be a potent muscle relaxant with intermediate duration of action. It had no cumulative effects even after as many as five maintenance doses. Doses of 0.4 mg/kg provided ideal intubating conditions in 2 min, though maximal blockade only developed in about 6 min. Duador was easily antagonized by edrophonium and no signs of recurarisation have been observed. The initial dose of Duador caused a mean 28% increase in heart rate. The increase in heart rate was inversely proportional to the control rate. Repeat doses did not cause further elevation in frequency. Duador had no effect on mean blood pressure. PMID- 6299131 TI - Fluorescent nucleotide triphosphate substrates for snake venom phosphodiesterase. PMID- 6299132 TI - Quantitative determination of myeloperoxidase using tetramethylbenzidine as substrate. PMID- 6299133 TI - Dual-beam difference spectroelectrochemistry. PMID- 6299134 TI - Electrophysiological studies of the cutaneous innervation of the external genitalia of the male dog. PMID- 6299135 TI - Twitch depression and train-of-four ratio after antagonism of pancuronium with edrophonium, neostigmine, or pyridostigmine. AB - During N2O-O2-halothane anesthesia pancuronium (3 mg/70 kg) was antagonized with neostigmine (2.5 or 5 mg/70 kg), pyridostigmine (10 or 20 mg/70 kg), or edrophonium (50 or 100 mg/70 kg) in 36 human subjects (6 in each group). Reversal was attempted at 10% spontaneous recovery of muscle twitch, which was measured using train-of-four stimulation. When first twitch tension was less than 70% of the control it was found that for the same tension, the train-of-four ratio was greater with edrophonium than with neostigmine, and greater with neostigmine than with pyridostigmine. It was concluded that the three antagonists have different mechanisms of action. In comparison with neostigmine, edrophonium is more and pyridostigmine is less effective at presynaptic (or fade) receptors. PMID- 6299136 TI - Phosphorylase ratio and susceptibility to malignant hyperthermia. AB - The ratio of muscle phosphorylase a to total phosphorylase, expressed as a percent, was determined in vastus lateralis muscle of 26 patients to examine the efficacy of this parameter as a method for screening for susceptibility to malignant hyperthermia (MH). As standard screening, all patients also had muscle contracture responses determined to 2% halothane and 0.25-32 mM caffeine at 37 degrees C. Each drug was given separately and not combined. Nine patients were susceptible to MH, based upon caffeine threshold of 2 mM or less (seven patients) or a rapidly developing contracture tension to halothane of more than 400 mg (seven patients, including five with positive caffeine responses). Mean phosphorylase ratio in these nine patients was 14.5 +/- 2.0% (mean, SEM). In the 17 nonsusceptible patients mean phosphorylase ratio (12.4 +/- 1.9%) was not significantly different. The range of phosphorylase ratios in susceptible patients was 6.5-26% while 13 nonsusceptible patients had ratios greater than 6% and up to 29%. The unacceptably high number of false-positive responses in nonsusceptible patients precludes the use of phosphorylase ratio as a definitive diagnostic test. PMID- 6299137 TI - Caprine arthritis-encephalitis: clinicopathologic study. AB - Chronic arthritis caused by caprine arthritis-encephalitis virus was observed after the introduction of new animals into a goat herd. There were high frequency of carpal hygroma and clinical signs of stiffness. The disease was progressive and produced a debilitating lameness among 30% of the affected animals. Laboratory findings were limited to alterations in synovial fluid which showed increased numbers of lymphocytes. Pathologic changes were observed in the joints, bursae, and adjacent tissues. Vascular injury and capillary leakage resulted in exudation into synovial cavities. Fibrin coated the synovial lining and formed (amorphous) long thread-like or broad-based villi. The articular cartilage was eroded. Cartilage erosion and penetration of the articular cartilage by pannus were associated with the presence of subchondral pseudocysts. The morphologic changes in bone and synovial tissues were like those described in human rheumatoid arthritis, except that rheumatoid nodules were not observed. PMID- 6299138 TI - Experimentally induced parainfluenza type 3 virus infection in young lambs: pathologic response. AB - Pulmonary changes in five 1-week-old, colostrum-deprived lambs transtracheally inoculated with parainfluenza type 3 virus were studied by immunofluorescent, microscopic, and ultrastructural techniques. The lambs were killed at postinoculation days (PID) 3, 5, and 7. Immunofluorescence specific for parainfluenza type 3 virus was first seen in small airways and alveolar epithelium and later in the lumens of airways and alveoli and, to a lesser extent, in the interstitium of the lungs. Grossly, there were multifocal areas of consolidation in all lobes of the lungs. These areas were characterized microscopically by bronchiolitis and interstitial pneumonitis. The bronchiolitis involved the terminal airways and consisted of necrosis and sloughing of epithelial cells followed by hyperplasia of the epithelium. The interstitial lesion comprised extensive infiltration of alveolar septa and alveoli with macrophages and the necrosis of alveolar epithelium. This was followed by hyperplasia of the epithelium. Degenerated bronchiolar and alveolar epithelium contained numerous intracytoplasmic inclusions early in the infection, but such inclusions were not seen in the lambs killed at PID 7. The degenerated changes were also seen with the electron microscope, as were numerous inclusions of viral nucleoprotein and a few viral buds at PID 3 and 5. Viral inclusions and buds were seen in ciliated and nonciliated bronchial epithelial cells and type I and type II alveolar epithelial cells. PMID- 6299139 TI - Immune response of pigs inoculated with virulent pseudorabies virus and pigs inoculated with attenuated or inactivated pseudorabies virus vaccine before and after challenge exposure. AB - Pseudorabies virus (PRV) antibodies, detectable by indirect radioimmunoassay (IRIA), serum-virus neutralization test (NT), or microimmunodiffusion test (MIDT) were developed within 8 days after pigs were inoculated with virulent PRV or attenuated PRV vaccine. Indirect radioimmunoassay and NT titers in pigs inoculated with virulent PRV were developed at the same rate, with IRIA titers being higher than NT titers. Pigs inoculated with attenuated or inactivated PRV vaccine developed peak mean prechallenge NT antibody titers of 4 and 1 (reciprocals of serum dilutions), respectively. Pigs inoculated with attenuated PRV vaccine had peak mean prechallenge IRIA antibody titers of 6, whereas pigs inoculated with inactivated PRV vaccine had mean IRIA antibody titers of 64. Challenge exposure of swine inoculated with attenuated or inactivated PRV vaccine elicited quantitatively equivalent responses, as measured by IRIA or NT, which were higher than prechallenge titers. There were no false-positive IRIA, NT, or MIDT results obtained when sera from nonvaccinated, nonchallenge-exposed pigs were tested. It appears that the PRV infection status of a seropositive swine herd could be ascertained by serologically monitoring several representative animals from a herd, using the NT. If 2 or more tests of representative animals at 14-day intervals were done and the mean NT titer was 4 or less, it could be concluded that the herd was vaccinated against, but not infected with, virulent virus. PMID- 6299140 TI - Evaluation of a killed feline panleukopenia virus vaccine against canine parvoviral enteritis in dogs. AB - Immunogenic potency of a killed feline panleukopenia virus vaccine against canine parvoviral enteritis in dogs was examined. The vaccine elicited hemagglutination inhibition antibodies to canine parvovirus (CPV) in all of the 72 dogs which were vaccinated. The vaccine was protective in dogs against both experimentally induced and naturally occurring CPV-induced disease. By statistical analysis, 4 weeks was found to be the optimal spacing between 2 vaccinal doses resulting in hemagglutination-inhibition antibody titers up to 1:5,120. Adverse reactions to the vaccine were not observed. Atypical lymphocytes were found consistently in the CPV-infected control dogs. PMID- 6299141 TI - Papillomatosis of the bovine teat (mammary papilla). AB - A 4th of 667 cattle examined at a Wisconsin abattoir had teat papillomas. Excised teat papillomas were sorted by gross morphologic characteristics into 3 groups: (i) atypical filiform, (ii) atypical flat, and (iii) typical fibropapilloma. Bovine papilloma virus capsid antigen was detected in thin-section slides of the 3 groups of teat papillomas by peroxidase-antiperoxidase assay. The bovine papilloma virus involved with the atypical papillomas could not be characterized by molecular hybridization, because enough pure virus could not be harvested. Homogenates of the 3 groups of teat papillomas were inoculated on 2 ponies and 4 calves. Typical fibropapillomas were produced on the 4 calves, and fibromas, on the 2 ponies. Atypical papillomas were produced only in 2 heifers. PMID- 6299142 TI - Use of modified live feline panleukopenia virus vaccine to immunize dogs against canine parvovirus. AB - Modified live feline panleukopenia virus (FPLV) vaccine protected dogs against canine parvovirus (CPV) infection. However, unlike the long-lived (greater than or equal to 20-month) immunity engendered by CPV infection, the response of dogs to living FPLV was variable. Doses of FPLV (snow leopard strain) in excess of 10(5.7) TCID50 were necessary for uniform immunization; smaller inocula resulted in decreased success rates. The duration of immunity, as measured by the persistence of hemagglutination-inhibiting antibody, was related to the magnitude of the initial response to vaccination; dogs with vigorous initial responses resisted oronasal CPV challenge exposure 6 months after vaccination, and hemagglutination-inhibiting antibodies persisted in such dogs for greater than 1 year. Limited replication of FPLV in dogs was demonstrated, but unlike CPV, the feline virus did not spread to contact dogs or cats. Adverse reactions were not associated with living FPLV vaccination, and FPLV did not interfere with simultaneous response to attenuated canine distemper virus. PMID- 6299143 TI - Attempted immunization of cats against feline infectious peritonitis, using avirulent live virus or sublethal amounts of virulent virus. AB - Kittens vaccinated with an avirulent biotype of the Black strain of feline infectious peritonitis virus (FIPV; given oronasally) developed both indirect fluorescent and virus-neutralizing antibodies, but were not protected against oronasal challenge exposure with virulent virus. In fact, kittens vaccinated with avirulent virus were more readily infected than were nonvaccinated cats. A proportion of kittens could be immunized to FIPV by giving sublethal amounts of virulent virus. This technique, however, was too inconsistent and hazardous to have clinical relevance. The results of these studies indicated that humoral immunity was not protective in FIPV infection. There was no correlation between fluorescent and virus-neutralizing antibodies and either disease or immunity. Immune serum from FIPV-resistant cats failed to passively protect susceptible animals against virulent virus given intraperitoneally or oronasally, and as expected, actually sensitized them to infection. It was concluded that cell mediated immunity was probably responsible for protection. PMID- 6299144 TI - Analysis of the genomes of bluetongue virus serotype 10 vaccines and a recent BTV 10 isolate from Washington. AB - The 10 double-stranded RNA gene segments of 2 vaccinal strains of bluetongue virus (BTV) serotype 10 that are used in the United States (BTV CA8 California and BT-8 Colorado), and a BTV-10 isolate recently obtained from infected sheep in Washington (state) were characterized by oligonucleotide fingerprint analyses. It was determined that although the 2 BTV-10 vaccinal strains are genotypically distinct, they are closely related both to each other and to the United States prototype BTV-10 virus. The BTV-10 field isolate appears to be a naturally occurring reassortment virus with genome segments derived from both United States prototype BTV-10 and BTV-11 viruses. However, one RNA segment of the isolate was totally unlike the corresponding segments of United States prototype BTV-10, -11, -13 and -17 viruses. PMID- 6299145 TI - Molecular epizootiologic studies of equine herpesvirus-1 infections by restriction endonuclease fingerprinting of viral DNA. PMID- 6299146 TI - Experimental latent and recrudescent bovine herpesvirus-1 infections in calves. AB - Latent bovine herpesvirus-1 (BHV-1) infection was established in 6 calves and was demonstrated by reinduction of virus shedding after administration of corticosteroids. Latently infected calves failed to transmit BHV-1 during 4 weeks' contact with sentinel calves. Infected calves were killed and necropsied during latency or induced recrudescence. The BHV-1 DNA was demonstrated intranuclearly in trigeminal ganglion neurons by in situ hybridization. The BHV-1 antigen was demonstrated by immunofluorescence in trigeminal ganglion neurons during recrudescence. By electron microscopy, changes in the appearance of the Nissl bodies and a high frequency of nuclear bodies were observed in trigeminal ganglion neurons. PMID- 6299147 TI - Modulation of early cellular events in wound healing in mice. AB - Wound healing experiments were conducted in random-bred Swiss mice to determine the effect on antimicrobial surfactants and macrophage stimulators on wound measurements, histologic repair, and wound breaking strengths. In selection of mice, Sendai virus antibody-positive mice healed more slowly than did mice with no detectable titer to Sendai virus. Studies were conducted with Sendai virus free mice that had C31G (an antimicrobial surfactant), alkyl amine oxide, zymosan, glucan, or phosphate-buffered isotonic saline solution instilled into full-thickness incised wounds. The early events in the repair process indicated a greater degree of inflammatory response comprised mainly of polymorphonuclear leukocytes with subsequent large numbers of monocytes in C31G and alkyl amine oxide-treated wounds. Although zymosan did not induce as large a number of monocytes, the degree of fibroplasia was as great as in wounds in which numbers were higher. The effect of zymosan could be blocked by the addition of N-alpha-p tosyl-L-lysine chloromethyl ketone to wounds. Wound breaking strength 3 days after surgery was greatest for glucan-treated mice (134 +/- 37 g) whereas that in C31G-treated mice (77 +/- 31 g) was less than that of the controls (92 +/- 37 g). By day 7, there was no significant difference in breaking strength between control and glucan-treated wounds; however, C31G-treated wounds remained substantially weaker than control wounds. PMID- 6299148 TI - Management of fungal diseases. AB - Diseases caused by the normally pathogenic fungi are common. The vast majority of patients will not require treatment. Amphotericin B remains the gold standard when treatment is needed. The availability of ketoconazole has provided an alternative mode of therapy, but its exact place is not yet defined. PMID- 6299149 TI - Electrical sizing of particles in suspensions. V. High electric fields. PMID- 6299150 TI - Membrane changes induced by high electric fields evidence for sulfhydryl group involvement. PMID- 6299151 TI - Opportunistic infections and Kaposi's sarcoma among Haitians: evidence of a new acquired immunodeficiency state. AB - Twenty Haitian patients, hospitalized from 1 April 1980 to 20 June 1982, had Pneumocystis carinii pneumonia, central nervous system toxoplasmosis, esophageal candidiasis, cryptococcosis, disseminated cytomegalovirus, progressive herpes simplex virus, chronic enteric coccidiosis, or invasive Kaposi's sarcoma. Ten patients died. Opportunistic infections were frequently multiple and were recurrent in three patients. In seven patients disseminated tuberculosis preceded the other infections by 2 to 15 months. There was no evidence of an underlying immunosuppressive disease, and no history of homosexuality or intravenous drug abuse. At least three patients probably acquired the syndrome in Haiti. Lymphadenopathy was common. Seventeen patients tested had anergy, and 18 had lymphopenia. Monoclonal antibody analysis of peripheral-blood T-cell subsets done on 11 patients showed a marked decrease in T-helper cells and an inversion of the normal ratio of T-helper cells to T-suppressor cells. This syndrome among heterosexual Haitians is strikingly similar to the syndrome of immunodeficiency described recently among American homosexuals. PMID- 6299152 TI - Acquired immunodeficiency syndrome in a patient with hemophilia. AB - A patient with hemophilia A developed T-cell deficiency characterized by infection with several opportunistic pathogens. Immunologic investigation showed cutaneous anergy, lymphocyte unresponsiveness to mitogens and antigens, an abnormal ratio of T-helper and T-suppressor cells with absolute lymphopenia and elevated IgA. The clinical and immunologic characteristics of this patient fit the recently described syndrome of opportunistic infections or Kaposi's sarcoma in patients with acquired T-cell deficiency; however, this patient does not have any of the associated underlying risk factors such as homosexuality, intravenous drug or amyl nitrite use, or positive serologic tests for syphilis. We conclude that the patient's acquired T-cell deficiency can be explained by exposure to a virus or other transmissible agent during factor VIII transfusions. PMID- 6299153 TI - Acquired immunodeficiency syndrome with Pneumocystis carinii pneumonia and Mycobacterium avium-intracellulare infection in a previously healthy patient with classic hemophilia. Clinical, immunologic, and virologic findings. AB - A previously healthy patient with classic hemophilia who was on a home infusion program with factor VIII concentrates developed an acquired immunodeficiency syndrome manifested by a dramatic weight loss (47 kg over 12 months), lassitude, transient thrombocytopenia, and opportunistic infections with Varicella zoster, Pneumocystis carinii, and Mycobacterium avium-intracellulare. The patient was not homosexual and had no history of intravenous drug abuse. Immunologic studies showed a persistent lymphopenia with reversal of helper/suppressor-cytotoxic T lymphocyte ratios, depression of human natural killer cell function, and in-vitro lymphocyte proliferative responses to mitogens and viral antigens. Serum IgA levels were also elevated. Serum antibodies against cytomegalovirus, herpes simplex viruses 1 and 2, Epstein-Barr virus, Varicella zoster, and hepatitis B virus were shown, suggesting previous infection by these agents. Reactivation of cytomegalovirus infection was suggested by a rising titer of antibodies against cytomegalovirus concurrent with pneumocystis pneumonia, and was confirmed by the growth of this virus in a throat culture 2 months later. PMID- 6299155 TI - Resistance of herpes simplex to acyclovir. PMID- 6299154 TI - The acquired immunodeficiency syndrome and Mycobacterium avium-intracellulare bacteremia in a patient with hemophilia. AB - A 27-year-old previously healthy man with hemophilia presented with Pneumocystis carinii pneumonia. The patient had several episodes of oral candidiasis followed by disseminated infection with Mycobacterium avium-intracellulare. He was not homosexual nor did he take illicit drugs, but he had been self-administering two to four monthly infusions of factor VIII concentrate for 7 years. In-vitro lymphocyte studies showed findings consistent with the acquired immunodeficiency syndrome that had previously been reported only in homosexual men, drug addicts, and Haitian refugees. The cause of this syndrome is unknown, but the possibility that it is associated with a transmissible agent acquired through the use of blood products such as factor VIII concentrate must be considered. PMID- 6299156 TI - The enzymatic profile of Haemophilus ducreyi. AB - The enzymatic activities of two reference strains of Haemophilus ducreyi and thirty clinical isolates were investigated by conventional biochemical tests and the API-ZYM test kit system which included 97 synthetic substrates. No strains converted delta-aminolevulinic acid to porphyrins, but they all reduced nitrates to nitrites. All strains possessed aminopeptidase activity against beta naphthylamide derivatives of L-alanine, L-arginine, L-glutamine, glycine, L leucine, L-lysine and L-serine. No trypsinor chymotrypsin-like activities were detected. All strains had phosphatase activity with broad pH range, and phosphoamidase activity. No glycosidase was detected by the substrates tested. PMID- 6299157 TI - Membrane IgD expression and dynamics in clones of human B-lymphoblastoid cells. PMID- 6299158 TI - Quartz: correlation between crystallinity index and fibrogenicity? PMID- 6299159 TI - Use of steroid/antibiotic prophylaxis in intraocular lens implantation: a double masked study v placebo. AB - A prospective, randomized, double-masked study was conducted to compare the efficacy of steroid/antibiotic treatment to placebo in reducing postoperative ocular inflammation. Patients undergoing planned extracapsular cataract extraction with intraocular lens implantation were treated with either placebo or a combination product containing dexamethasone, neomycin, and polymyxin B for three days prior to surgery and three weeks following surgery. Five of thirteen (38%) patients treated with placebo developed significant iritis postoperatively, while none of ten (0%) patients treated with the combination product developed excessive inflammation (P = 0.027). Two patients treated with the steroid/antibiotic combination product experienced a broken suture postoperatively followed by iris prolapse, as compared to none in the placebo group (P greater than 0.10). More types and significant numbers of bacteria were isolated from the eyes of the placebo-treatment group after therapy than were found in the steroid/antibiotic-treatment group. PMID- 6299160 TI - Irreplaceable free flaps in reconstructive surgery--part I. AB - Free flaps are increasingly being chosen to solve complex reconstruction problems. We describe a fascial sleeve approach to elevation of the donor flap which increases its safety and reliability. We report examples in which resultant revascularization of the recipient site causes the free flap to be irreplaceable for solving the particular clinical problem. PMID- 6299161 TI - Factors influencing wound complications: a clinical and experimental study. AB - Burst abdomen, incisional herniation, sinus formation and post-operative wound infection continue to bedevil the surgeon. A prospective study of 1129 laparotomy wounds defined the extent of the problem; 1.7% incidence of dehiscence, 7.4% herniation and 6.7% sinus formation, all significantly associated with wound infection. Mass closure reduced the dehiscence rate from 3.8% to 0.76%. Infection reduced wound strength in a rat laparotomy model due to a decrease in fibroblast concentration and activity. A monofilament non-absorbable suture was shown experimentally to be the most suitable suture for closing infected abdominal wounds. Electron microscopy demonstrated bacteria in the interstices of infected multifilament sutures. A randomised clinical trial comparing polyglycolic acid and monofilament nylon in the closure of abdominal wounds confirmed the experimental findings; polyglycolic acid resulted in a significantly higher wound failure rate with no decrease in sinus formation. A mass closure technique using monofilament nylon is recommended for laparatomy closure and efforts should continue to reduce wound sepsis. PMID- 6299162 TI - [Granular cell tumor of the biliary tract. Apropos of a case]. PMID- 6299163 TI - Clinical applications of prostaglandins in Obstetrics and Gynaecology. AB - Although a number of potential practical uses of prostaglandins have been identified, these compounds have so far found clinical applications mainly in Obstetrics and Gynaecology. It is almost 15 years since a prostaglandin was first used for the induction of term labour and prostaglandin E2 is now commercially available for this purpose in many countries. For the termination of second trimester pregnancy, prostaglandins have almost completely replaced other methods previously in use. Other areas where prostaglandins are routinely used or where their uses are being developed, include menstrual induction, preevacuation dilatation of the cervix in the first trimester, termination of pregnancy in cases of missed abortion, intrauterine fetal death and other types of abnormal pregnancies, control of post-partum haemorrhage, treatment of post-partum or post surgical urine retention and ripening of the cervix prior to induction of labour at term. In early studies, prostaglandins E2 and F2 alpha were used for all the applications listed above. In order to increase efficacy and reduce side effects, a number of synthetic analogues were later evaluated for application in selected areas. Those with modification in the 15 and 16 positions of PGE2 and PGF2 alpha molecules have undergone extensive clinical trials and some of these analogues are now in routine use. The current status of the practical applications of prostaglandins in Obstetrics and Gynaecology is reviewed. PMID- 6299164 TI - The diagnosis of gestational trophoblastic disease. PMID- 6299166 TI - HLA and trophoblastic disease. AB - The susceptibility of trophoblastic diseases is not associated with HLA antigens. Compatibility studies between the patient and her husband, and the patient and the tumour suggest that it may not play an important role in the susceptibility or the progression of the tumour. This as well as other direct evidence indicate that the tumour cells probably do not express much HLA antigens on their cell surface. The expression of HLA antigens on normal and abnormal trophoblasts are discussed. PMID- 6299165 TI - Recent advances in the management of gestational trophoblastic disease. AB - What was once an almost uniformly fatal disease has now become highly curable, with survival rates of 100% being reported for the early stages of the disease. This is the direct result of improved techniques of diagnosis, recognition of high risk groups, effective and intensive chemotherapy, adjunctive surgery in selected cases and most important of all, a close follow-up of all patients with monitoring of the tumour marker, human chorionic gonadotrophin (hCG). This review will highlight the advances made in these areas and how these have helped in reducing morbidity and mortality from gestational trophoblastic disease (GTD). PMID- 6299167 TI - Review on coagulation in trophoblastic disease. AB - Problems of haemostasis associated with abnormal pregnancy states such as eclampsia, prolonged intrauterine death and amniotic fluid embolism causing maternal death are well recognised. Coagulation problems have been suspected in hydatidiform molar pregnancies previously but because of the rare occurrence, the importance of defective haemostasis in this trophoblastic disorder has not been emphasised. Evidence from clinical course of the disease, from haematological studies and from postmortem findings provide evidence that enhanced coagulation occurs in all hydatidiform molar pregnancies. Secondary fibrinolysis in the intact mole increases during evacuation and could aggravate the coagulation state. If there should already be a moderate degree of intravascular coagulation, it could progress to an irreversible state with evacuation if the coagulation defect are not corrected before the procedure as illustrated by reported maternal deaths. Since the degree of vaginal bleeding does not definitely indicate the degree of coagulopathy, coagulation screening tests are necessary to detect the condition in all hydatidiform moles. The placental proteins with coagulation and fibrinolytic activity found in trophoblastic disease, may be associated with the aetiology of coagulation disorder and may have a prognostic value in detecting subsequent malignancy. PMID- 6299168 TI - [Immunology of beta-adrenergic receptors: a working model of a hormone-mimicking internal image]. AB - Two types of antibodies have been prepared against beta-adrenergic receptors: (1) antibodies against receptor, purified from turkey erythrocyte membranes by affinity chromatography on alprenolol sepharose, and (2) antiidiotypic antibodies raised against the anti-alprenolol immunoglobulins. Both types of antibodies specifically bind to cells which possess beta-adrenergic receptor and mimick the biological effect of the catecholamine hormone: they stimulate basal adenylate cyclase and enhance adenylate-cyclase activation by catecholamines. The antiidiotypic antibodies only compete with (--)-3H-dihydroalprenolol for binding to the beta-adrenergic receptors on purified turkey erythrocyte membranes. The use of these antibodies as tools for the study of the mechanism of the beta adrenergic receptor-cyclase complex is discussed. PMID- 6299169 TI - On double-minutes and their origin in a benign human neoplasm, a mixed salivary gland tumour. AB - The cytogenetical observations in the first double-minute (DM)-positive benign neoplasm, a human pleomorphic adenoma, are described. It is suggested that in the present adenoma the first DM was formed from a structurally and/or functionally specialized segment of the long arm of a chromosome No 12. If so, the results also indicate a close relation between chromosome regions preferentially involved in structural rearrangements in a certain tumour type and chromosome regions with the potential capacity to become released and behave and function as DMs. PMID- 6299171 TI - Natural antibodies in human sera cross-reactive with type C RNA tumor viral P 30. AB - Antibodies cross-reactive with type C viral p30 were concentrated from 105 samples of human plasma by affinity chromatography using Sepharose beads to which disrupted Simian sarcoma associated virus (SSAV) had been coupled. Eluates obtained from the affinity beads were tested for anti-p30 activity by performing radio-immunoprecipitations with 125I-labeled SSAV p30 and Rauscher murine leukemia viral (R-MuLV) p30. Forty six percent of the eluates were found positive when tested against SSAV p30. From 44 positive eluates for anti SSAV p30 activity, 20 (45%) were also found to be positive for anti-R-MuLV p30 activity. No significant difference was found in anti-p30 activity in eluates from normal controls and from patients with various kinds of malignancy. PMID- 6299173 TI - Changes in the protein composition of Epstein-Barr Virus-negative lymphoma cells after EBV-conversion. AB - Two-dimensional protein separation, combining high resolution isoelectric focussing with SDS gel electrophoresis, was applied to analyse Epstein-Barr Virus (EBV)-induced changes in the protein composition of lymphoma cells. Total lysates of the EBV-negative Burkitt lymphoma lines Ramos and BJAB were compared with their permanently converted EBV-genome positive sublines, after 35S-methionine labelling. Five out of seven EBV-infected lines showed a 68k protein, which was absent from both EBV-free parental lines, and was identified as a glycoprotein (gp 68) by metabolic sugar labelling. We conclude that presence of this protein is a direct or indirect consequence of the presence of the EBV genome. PMID- 6299172 TI - Effect of dimethylnitrosamine on the concentration of polyamines in rat liver and kidney. AB - Newborn female rats were injected once with dimethylnitrosamine. The polyamine content of the liver and kidneys was measured and compared to the respective controls. Both liver and kidney polyamine concentrations were found to decrease in both organs up to one month of age. The polyamine concentrations then increased in the carcinogen treated animals, whereas the control levels remained unchanged. The concentration of polyamines was greater in the left kidney than in the right one. A close correlation was found between the concentration of polyamines and histological findings during tumour development. PMID- 6299170 TI - Monovalent cation transport during the cell cycle (review). AB - This paper reviews the properties and regulation of cation transport during the cell cycle. Extensive modulations occur in both electro-diffusional and non electro-diffusional cation transport, most prominently during mitosis and G1 phase. These modulations can be related to compositional, structural and dynamic membrane properties. The implications of modulated cation transport for progression through the cell cycle are discussed. PMID- 6299174 TI - DNA ploidy in small cell carcinoma of the lung. AB - Fluorometric single cell DNA analysis was performed on 123 tumour samples from 33 patients who had died of small cell carcinoma of the lung (SCCL). In 36% (12 patients) the modal DNA profile was hypodiploid or near diploid and the median survival time was 16.3 months, as opposed to 9.5 months for the 21 patients with hyperdiploid DNA profile, who also seemed to have a lower response rate to chemotherapy. It is suggested that an estimation of the modal DNA ploidy before the start of the treatment in SCCL might be used as a cellular parameter to obtain more comparable treatment study groups and for the prediction of survival. PMID- 6299175 TI - The pathophysiology of brain ischemia. AB - Brain ischemia due to a critical reduction in cerebral blood flow is a well recognized and common cause of irreversible brain damage. The observation that brain cells are more resistant to ischemia than was previously assumed on the basis of clinical experience has stimulated considerable investigative work designed to determine those factors responsible for irreversible ischemic cell damage. At this time, data from these investigations indicate that cellular acidosis and biochemical disturbances initiated by abnormal intracellular ion homeostasis may be especially important in determining the ultimate survival of nerve cells. This review examines the biochemical events initiated by ischemia and their potential role in determining the ultimate survival of brain cells. PMID- 6299176 TI - Syndrome of opsoclonus-myoclonus caused by Coxsackie B3 infection. AB - Coxsackie B3 was cultured from 2 children who presented within two weeks of each other from the Cape Cod, Massachusetts, area showing opsoclonus and myoclonus. The organism was recovered from the cerebrospinal fluid of 1 patient and from the stools of both. Both children had cerebrospinal fluid pleocytosis and gradual, spontaneous resolution of their symptoms. PMID- 6299177 TI - Hormonal influences on memory. PMID- 6299178 TI - Effects of pH on the activity of ketoconazole against Candida albicans. AB - The activity of ketoconazole against Candida albicans in Sabouraud glucose medium was markedly influenced by pH. Minimum inhibitory concentrations of this imidazole against both a standard strain and clinical isolates ranged from 40 mug/ml at pH 3 to 0.02 mug/ml at pH 7, a greater than 1,000-fold difference. PMID- 6299179 TI - Inhibition of human cytomegalovirus by trifluorothymidine. AB - The antiviral activity of trifluorothymidine (TFT) singly and in combination with other antiviral agents against human cytomegalovirus (HCMV) was evaluated by using an infectious center plaque reduction assay. The 50% inhibitory dose of TFT against six different patient HCMV strains was 0.57 (+/- 0.24, standard deviation) microM and ranged from 0.32 to 0.97 microM. The 50% inhibitory dose for the laboratory-adapted HCMV strain, AD-169, was 2.1 microM. When TFT (0.17 microM) was combined with human fibroblast interferon (25 U/ml), the combination was additive against all four HCMV isolates evaluated. Synergism was observed when TFT (0.17 microM) was combined with phosphonoformic acid (25 microM) for all strains studied or with acyclovir (20 microM) for three of the four clinical HCMV strains tested. Each of the three antiviral agents, when combined with TFT, exhibited additive effects against strain AD-169. TFT at concentrations of 0.5, 1.7, and 3.5 microM had an increasing inhibitory effect on uninfected human embryonic lung fibroblast (HEL) cell growth over 72 h, with 16% growth inhibition at 3.5 microM after 3 days. There was no increased toxicity to growing HEL cells when the paired antiviral agent combinations were evaluated. These findings suggest that TFT may be useful singly or in combination with other antiviral agents in treating HCMV infections. PMID- 6299180 TI - Combined effect of acyclovir and amphotericin B on the replication of pseudorabies virus in BHK-21 cells. AB - Acyclovir, known as an antiherpetic agent, showed an inhibitory effect on the propagation of pseudorabies virus in BHK-21 cells. The antiviral effect of acyclovir was observed by plaque reduction, as well as by the inhibition of the virus-stimulated uptake of thymidine by BHK-21 cells. Amphotericin B potentiated the antiviral activity of acyclovir. The optimal concentrations of polyene antibiotic expressing the potentiating effect were lower than required for the induction of K+ leakage from the cells. There was no evident amphotericin B induced stimulation of thymidine incorporation into infected BHK-21 cells. The model presented may be useful to study the potentiation phenomenon of polyene macrolide antibiotics. PMID- 6299181 TI - Toxicity of amphotericin B, miconazole, and ketoconazole to human granulocyte progenitor cells in vitro. AB - Granulocyte progenitor cells were grown in culture with amphotericin B, miconazole, and ketoconazole. Significant suppression of progenitor cell growth could be demonstrated with all three drugs at increasing concentrations. No additive suppression was seen when amphotericin B and ketoconazole were combined. PMID- 6299182 TI - Effect of ketoconazole on the fungicidal action of amphotericin B in Candida albicans. AB - Amphotericin B-susceptible Candida albicans became resistant to the drug after growth in the presence of ketoconazole. Chromatographic analysis of cellular sterols showed that the organisms became depleted of ergosterol in parallel with the development of amphotericin B resistance. The implications of these findings are discussed in relation to combination chemotherapy with these two important antifungal agents. PMID- 6299183 TI - Application of the fluorescamine reaction with 6-aminopenicillanic acid to estimation and detection of penicillin acylase activity. AB - 6-Aminopenicillanic acid may be quantitatively estimated by its reaction with fluorescamine in the concentration range of 1 to 10 micrograms/ml. The difference in reactivity between 6-aminopenicillanic acid and benzylpenicillin, which does not react with fluorescamine, can be used to determine penicillin acylase activity and obtain data on enzyme parameters and inhibitors. Unlike amino acids and peptides, 6-aminopenicillanic acid reacts strongly with fluorescamine at pH 4, an observation which can be used to determine the presence of penicillin acylase in whole bacterial cell preparations. PMID- 6299184 TI - Ceftriaxone diffusion into cerebrospinal fluid of children with meningitis. AB - We evaluated the diffusion of ceftriaxone into the cerebrospinal fluid of 27 infants and children with meningitis who were receiving conventional antimicrobic therapy. Ceftriaxone was administered as a single 75 mg/kg dose and was given early or late or both in the course of the illness. Three hours after a dose, the mean cerebrospinal fluid ceftriaxone level was 5.7 micrograms/ml in patients studied early in the course of meningitis and 2.1 micrograms/ml in patients studied later in the illness. Six hours after a dose, the mean cerebrospinal fluid ceftriaxone levels early and late in meningitis were 7.2 and 2.5 micrograms/ml, respectively. The diffusion did not correlate with the leukocyte count or the protein or glucose content of the cerebrospinal fluid. Serial, simultaneous cerebrospinal fluid and plasma ceftriaxone levels were also determined in three additional patients with ventriculo-peritoneal shunt infections and external ventriculostomy drainage. The cerebrospinal fluid ceftriaxone levels in these patients ranged from 0.7 to 8.3 micrograms/ml. Our data indicate that ceftriaxone diffuses sufficiently and consistently into the cerebrospinal fluid to warrant its assessment in the treatment of meningitis. PMID- 6299185 TI - Comparison study of the kinetics of ceftizoxime penetration into extravascular spaces with known surface area/volume ratio in vitro and in vivo in rabbits. AB - The extravascular kinetics of ceftizoxime were studied both in an in vitro kinetic model and in an in vivo rabbit model. Visking tubing chambers were used in both models to provide extravascular spaces with large or small volumes and surface areas, but identical surface area/volume ratios. Four rabbits, each implanted with two large Visking chambers and four small chambers, received 50 mg of ceftizoxime per kg intramuscularly every 3 h for eight doses. In the in vitro model, 80 mg of ceftizoxime was infused over 30 min every 3 h for eight doses. Intravascular and extravascular spaces were sampled in both models after the eighth dose. Ceftizoxime had similar intravascular kinetics in both models, i.e., the peak levels, the peak-to-trough fluctuations, and the half-life were comparable. The area under the curve (AUC) for the extravascular spaces was also similar in the two models. Large and small chambers having identical surface area/volume ratios demonstrated identical kinetics. The extravascular Visking chamber spaces achieved equilibrium with the intravascular spaces in both models, i.e., the AUC for the extravascular spaces was the same (P > 0.2) as that for the serum (rabbit model) or the test chamber (in vitro model). This study illustrates (i) that our modified in vitro model is a potentially valid model for studying extravascular kinetics; (ii) that extravascular spaces with identical surface area/volume ratios show similar penetration kinetics with a freely diffusible drug, such as ceftizoxime, despite differences in size; and (iii) that the Visking chamber extravascular-space model permits the free diffusion of the antimicrobial agent and reaches equilibrium (equivalent AUC) with the intravascular space. PMID- 6299186 TI - beta-Lactamase inhibitory activity of iodopenicillanate and bromopenicillanate. AB - Iodopenicillanate and bromopenicillanate were shown to be effective inhibitors of a variety of beta-lactamases. Staphylococcus aureus isolates were synergistically inhibited by iodopenicillanate and bromopenicillanate combined with ampicillin. Methicillin-resistant S. aureus was not synergistically inhibited. Escherichia coli which possessed TEM beta-lactamase activity had a reduction in ampicillin minimal inhibitory concentration, but an E. coli isolate which had chromosomal beta-lactamase and a low ampicillin minimal inhibitory concentration showed no reduction in ampicillin minimal inhibitory concentration with either iodopenicillanate or bromopenicillanate. Of the Klebsiella isolates tested, 80% were synergistically inhibited by both iodopenicillanate and bromopenicillanate. Most Morganella isolates were synergistically inhibited by either iodopenicillanate or bromopenicillanate combined with ampicillin, as were many Salmonella, Shigella, Citrobacter diversus, and Citrobacter freundii isolates. No Pseudomonas aeruginosa isolates were synergistically inhibited by iodopenicillanate or bromopenicillanate. Preincubation did not improve the inhibitory activity of iodopenicillanate or bromopenicillanate. PMID- 6299187 TI - Synthesis of some 5'-amino-2',5'-dideoxy-5-iodouridine derivatives and their antiviral properties against herpes simplex virus. AB - In an attempt to improve the antiviral efficacy of 5'-amino-2',5'-dideoxy-5 iodouridine (AIdU) the N-acetyl and N,3'-O-diacetyl derivatives were prepared. N Acetylation of AIdU increased its ability to inhibit the phosphorylation of thymidine by the deoxypyrimidine kinase of herpes simplex virus type 1 (HSV1) while diacetylation had the converse effect. The affinity of the corresponding compounds containing uracil or thymine for virus deoxypyrimidine kinase was also determined. A range of N-acyl-, N-sulphonyl- and N,3'-O-diacyl- derivatives of AIdU were synthesized; enhanced inhibition of deoxypyrimidine kinase by a number of these compounds was observed. The previous observation that 5'-azido-2',5' dideoxy-5-iodouridine has antiherpetic activity in vivo led us to investigate its 3'-O-acetyl derivative as well as the corresponding compound containing uracil. None of the derivatives described showed antiviral activity in cell culture against HSV1; acylation failed to enhance the potency of AIdU against HSV1 in vivo. PMID- 6299188 TI - Antiviral efficacy of pyrazofurin against selected RNA viruses. AB - The antibiotic pyrazofurin, 3-(beta-D-ribofuranosyl)-4-hydroxypyrazole-5 carboxamide, markedly inhibited the in vitro replication of a number of RNA viruses including Rift Valley fever (RVF), Venezuelan equine encephalomyelitis (VEE), Sandfly, Pichinde, Lassa and LCM virus. Plaque formation was reduced by 80% or more with 2-10 micrograms/ml of pyrazofurin while 2 micrograms/ml reduced by 1000-fold the yield of Lassa and LCM virus in a yield reduction assay. In vivo, pyrazofurin failed to protect mice and guinea pigs against a lethal challenge with VEE and Pichinde virus, respectively. On the other hand, pyrazofurin caused a slight increase in the mean time to death of mice infected with RVF virus. PMID- 6299189 TI - Prevention of death in mice infected with coxsackievirus A16 using guanidine HCl mixed with substituted benzimidazoles. AB - A significant reduction in the death rate of infant mice infected with ten 50% lethal doses (LD50) of coxsackievirus A16 was observed when they were treated 58 h after infection with two injections of guanidine at 145 mg/kg per injection. Tremors occurred at this level but disappeared after treatment was discontinued. Tremors were apparent, but less severe at 97 mg/kg per injection and did not occur at 48 mg/kg per injection. No antiviral effect could be detected at either of these levels of guanidine. When an inactive level of guanidine (97 mg/kg per injection) was combined with 1.7 mg/kg per injection of LY122771-72, LY127123, or 2-(alpha-hydroxybenzyl)benzimidazole (HBB) and 17 mg/kg per injection of 2 guanidino-benzimidazole (GB), significant activity resulted with 2-8 treatments begun 58 h after infection. The same treatment schedule using 136 mg/kg per injection of LY122771-72, 90 mg/kg per injection of LY127123, 136 mg/kg per injection of HBB and 68 mg/kg per injection of GB produced no effect. Guanidine associated tremors were also enhanced by the addition of the substituted benzimidazoles. When guanidine was reduced to 48 mg/kg per injection, 34 mg/kg per injection of LY122771-72 was required to produce a significant reduction in the death rate and no tremors were observed. PMID- 6299190 TI - Effect of cyclosporin A on the production of interferon by human peripheral blood leukocytes in vitro. AB - Cyclosporin A (CsA) was assessed for its effect on the production of antiviral activity by human peripheral blood leukocytes (PBL). CsA markedly reduced the production of interferon-gamma (IFN-gamma) in response to stimulation with lectin mitogens, bacterial products, alloantigens, or Epstein-Barr virus (EBV) transformed lymphoblastoid cell lines (LCL). CsA-mediated suppression of IFN gamma secretion was dose-dependent and did not result from a shift of kinetics of the production of antiviral activity. The production of IFN-alpha in response to stimulation with Corynebacterium parvum (CP), viruses, and synthetic polynucleotides was not affected by the addition of CsA. These findings confirm earlier observations that CsA predominantly acts on T lymphocyte function. CsA may prove a valuable agent to study the role of IFN-gamma in the pathogenesis of virus-associated malignant lymphoproliferative disease. PMID- 6299191 TI - Influence of salts on electrostatic interactions between poliovirus and membrane filters. AB - Neither solutions of salts nor solutions of detergents or of an alcohol at pH 4 are capable of eluting poliovirus adsorbed to membrane filters. However, solutions containing both a salt, such as magnesium chloride or sodium chloride, and a detergent or alcohol at pH 4 were capable of eluting adsorbed virus. The ability of ions to promote elution of virus at low pH in the presence of detergent or alcohol was dependent on the size of the ions and the ionic strength of the medium. These results suggest that both electrostatic and hydrophobic interactions are important in maintaining virus adsorption to membrane filters. Hydrophobic interactions can be disrupted by detergents or alcohols. It appears that electrostatic interactions can be disrupted by raising the pH of a solution or by adding certain salts. Disruption of either electrostatic or hydrophobic interactions alone does not permit efficient elution of the adsorbed virus at low pHs. However, when both interactions are disrupted, most of the poliovirus adsorbed to membrane filters is eluted, even at pH 4. PMID- 6299192 TI - Hysteretic behavior of rat liver fructose 1,6-bisphosphatase induced by zinc ions. PMID- 6299193 TI - Comparison of detergent-solubilized and membrane-bound forms of kidney (Na + K) ATPase. PMID- 6299195 TI - Solvent proton relaxation studies of cytochrome c oxidase solutions. AB - The interaction of solvent water protons with the bound paramagnetic metal ions of beef heart cytochrome c oxidase has been examined. The observed proton relaxation rates of enzyme solutions had a negative temperature dependence, indicating a rapid exchange between solvent protons in the coordination sphere of the metal ions and bulk solvent. An analysis of the dependence of the proton relaxation rate on the observation frequency indicated that the correlation time, which modulates the interaction between solvent protons and the unpaired electrons on the metal ions, is due to the electron spin relaxation time of the heme irons of cytochrome c oxidase. This means that at least one of the hemes is exposed to solvent. The proton relaxation rate of the oxidized enzyme was found to be sensitive to changes in ionic strength and to changes in the spin states of the metal ions. Heme a3 was found to be relatively inaccessible to bulk solvent. Partial reduction of the enzyme caused a slight increase in the relaxation rate, which may be due to a change in the antiferromagnetic coupling between two of the bound paramagnetic centers. Further reduction resulted in a decreased relaxation rate, and the fully reduced enzyme was no longer sensitive to changes in ionic strength. The binding of cytochrome c to cytochrome c oxidase had little effect on the proton relaxation rates of oxidized cytochrome oxidase indicating that cytochrome c binding has little effect on solvent accessibility to the metal ion sites. PMID- 6299194 TI - Photoaffinity labeling of a microtubule-associated cyclic AMP-binding protein. PMID- 6299196 TI - Infrared and electron paramagnetic resonance study of nitrosyl(protoporphyrin IX dimethyl ester)iron(II) and its complexes with nitrogenous bases as model systems for nitrosylhemoproteins: effect of solvent polarity. AB - Infrared and electron paramagnetic resonance spectra of nitrosyl(protoporphyrin IX dimethyl ester)iron(II)(Fe(PPDME)(NO)) and its complexes with nitrogenous bases (N bases) such as imidazoles, pyridines, aliphatic amines, and anilines have been measured in various solvents. At room temperature, giso, Aiso, and nu NO values of five-coordinate Fe(PPDME)(NO) decreased with an increase in solvent polarity parameter ET, indicating the interaction between the solvent and the vacant axial coordination position. It has been found that the nu NO value of six coordinate species is very sensitive to the solvent polarity, while the giso value is less sensitive. The solvent effect on the equilibrium constants, which are evaluated from the intensity change of the NO stretching band for five- and six-coordinate species, is discussed. PMID- 6299197 TI - A family of phosphohydrolases from bovine intestinal mucosa: 5'-nucleotidase. AB - Bovine intestinal 5'-nucleotidase has been partially purified and characterized for comparison with two other phosphohydrolases from the same tissue, alkaline phosphatase and 5'-nucleotide phosphodiesterase, which are closely related structurally and mechanistically. Kinetic studies with a variety of nucleotides and phosphonate analogs show that, although 5'-nucleotidase is a monoesterase like alkaline phosphatase, it more closely resembles 5'-nucleotide phosphodiesterase in its high affinity and specificity for nucleotide binding. 5' Nucleotidase is bound very strongly by an affinity column containing a bound phosphonate analog of ADP but is not bound by an affinity column containing a non nucleotide phosphonate which selectively binds alkaline phosphatase. 5' Nucleotidase is strongly bound by immobilized antibodies prepared against 5' nucleotide phosphodiesterase, and is less strongly bound by immobilized antibodies prepared against alkaline phosphatase. We conclude that 5' nucleotidase is structurally more similar to 5'-nucleotide phosphodiesterase than to another monoesterase, alkaline phosphatase. PMID- 6299198 TI - Growth hormone receptors in lymphocytes of growth hormone-deficient children. AB - Human growth hormone was injected intravenously into 18 growth hormone-deficient children and growth hormone binding sites in lymphocytes were investigated. Fresh circulating lymphocytes had a low initial value for the binding of growth hormone to solubilized receptors (3.45 +/- 1.46%) but after growth hormone injection, the binding rapidly increased to 14.8 +/- 4.2% at 2 1/2 h and 8.7 +/- 1.8% at 5 h. The sharp increase in binding is due to increase in the number of binding sites. Two control children who received chorionic gonadotropin had no change in lymphocyte growth hormone receptors. The methodological differences between the present study and previous attempts to identify human growth receptors in lymphocytes were (1) lymphocytes were separated and disrupted with Triton X-100 as quickly as possible (to avoid error from receptor leaking out of the cell) and (2) the receptors were assayed at 2 1/2 h after growth hormone administration (previous studies were 12-24 h later). One possible explanation for the data is that growth hormone receptor from liver is taken up by lymphocytes and rapidly released again, thus, contributing to the hormonal receptor economy in humans. PMID- 6299199 TI - In vitro modulation of renal 25-hydroxyvitamin D3 metabolism by vitamin D3 metabolites and calcium. AB - It has been shown that 1,25-dihydroxyvitamin D3 (1,25-(OH)2-D3) and dietary Ca modulate renal metabolism of 25-hydroxyvitamin D3 (25-OH-D3) to 1,25-(OH)2-D3 and 24,25-dihydroxyvitamin D3 (24,25-(OH)2-D3) in the rat. However, it is not known if 1,25-(OH)2-D3 and Ca act directly on the kidney to modulate 25-OH-D3 metabolism or indirectly through other mechanisms, such as the modulation of parathyroid hormone secretion. Therefore, we have used isolated renal cortical slices from the rat to study the effect of 1,25-(OH)2-D3 and Ca in vitro on renal 25-OH-D3 metabolism. Incubation of renal slices from rats fed a vitamin D deficient, low-Ca diet with 50 nM 1,25-(OH)2-D3 for 3 h resulted in a significant decrease in 1,25-(OH)2-D3 production and a significant increase in 24,25-(OH)2-D3 production. Increasing media Ca concentration from 0.5 to 2.5 mM resulted in a significant decrease in 1,25-(OH)2-D3 production but no change in 24,25-(OH)2-D3 production. The inhibitory effect of 1,25-(OH)2-D3 was blocked by cycloheximide, but the inhibitory effect of Ca was not blocked by cycloheximide. Renal 1,25 (OH)2-D3 production was inhibited to a greater extent by incubation with 1,25 (OH)2-D3 and Ca together than by incubation with 1,25-(OH)2-D3 and Ca separately. These studies indicate that 1,25-(OH)2-D3 and Ca act directly on the kidney to modulate renal 25-OH-D3 metabolism. They also suggest that the mechanism of modulation is different for each agent. PMID- 6299200 TI - 5'-deoxy-5'-thioanalogs of adenosine and inosine 5'-monophosphate: studies with 5'-nucleotidase and alkaline phosphatase. AB - The interaction of 5'-deoxy-5'-thioadenosine 5'-monophosphate (A(S)MP) and 5' deoxy-5'-thioinosine 5'-monophosphate (I(S)MP) with snake venom, 5'nucleotidase, and calf intestinal mucosa alkaline phosphatase has been characterized. The substrates, A(S)MP and I(S)MP, are analogs of adenosine 5'-monophosphate and inosine 5'-monophosphate in which sulfur replaces oxygen as the bridge between the 5'-carbon of the ribose and the phosphorous. The P-S bond of both A(S)MP and I(S)MP was hydrolyzed by alkaline phosphatase producing the corresponding thionucleoside as a reaction product. The Km for A(S)MP was 270 microM and the V for alkaline phosphatase was 110 nmol/min/mg (8% of the V for AMP), whereas the corresponding values for I(S)MP were 300 microM and 530 nmol/min/mg protein, respectively. In contrast, 5'-nucleotidase did not catalyze hydrolysis of either A(S)MP or I(S)MP. A(S)MP and I(S)MP were competitive inhibitors of the 5' nucleotidase hydrolysis of AMP and IMP, respectively, with Ki values of 975 and 13 microM. Decreasing the pH of the reaction from 8.1 to 7.1 lowered the Ki for I(S)MP by 100-fold, to a value of 0.15 microM. PMID- 6299201 TI - (Na+,K+)-ATPase of mammalian brain: effects of temperatures on cation and ATP interactions regulating phosphatase activity. AB - The effects of temperature on interactions between univalent cations or ATP and the p-nitrophenylphosphatase activity associated with brain (Na+,K+)-ATPase were examined. The apparent affinity for K+ activation under conditions favoring the moderate affinity site was temperature dependent, increasing with decreasing temperature. A comparison of univalent cations showed that the negative apparent delta H and delta S for cation binding increased with increasing apparent cation affinity. In contrast to the case with the moderate affinity sites, apparent affinity for the high affinity K+ site was independent of temperature. As temperature decreased, properties of moderate affinity site binding approached those of the high affinity site. The temperature dependence of ATP inhibition was opposite to that for K+ activation, with positive apparent delta H and delta S. The apparent delta H and delta S for cation binding approached those for the overall conformational change to K+-sensitive enzyme as cation affinity increased. These data suggest that E2, the K+-sensitive form of (Na+,K+)-ATPase, is stabilized by forces that require a decrease in entropy, explaining the predominant existence of E1 at physiologic temperatures. A conformational change leading to stabilization of E2 at higher temperatures can be produced by binding of univalent cations to a moderate affinity, presumably intracellular, site. This effect is counteracted by ATP. ATP also appears to alter the selectivity of this site to favor Na+ over K+ binding. PMID- 6299202 TI - In vitro inhibitor studies of vitamin D 25-hydroxylase in rat liver microsomes. AB - The ability of four vitamin D analogs to inhibit the liver microsomal vitamin D 25-hydroxylase was determined. 19-Hydroxy-10(S),19-dihydrovitamin D3,25 fluorovitamin D3, 3 beta-hydroxy-9,10-seco-5,7,10(19)-choletrien-24-oic acid dimethylamide and 25-aza-vitamin D3 were competitive inhibitors with apparent KI values of 44, 137, and 870 nM, and 6.4 microM, respectively. The values for the 19-hydroxy-10(S), 19-dihydrovitamin D3, 25-fluorovitamin D3, and 25-aza-vitamin D3 correspond well to other literature reports with respect to their relative in vivo inhibitory properties. 24-Oxovitamin D3 oxime also proved to be a potent inhibitor but a detailed analysis was prohibited by the lack of material. The 3 beta-hydroxy-9,10-seco-5,7,10(19)-choletrien-24-oic acid dimethylamide was also tested in vivo but had no antagonistic activity when provided at a 2000-fold excess over vitamin D3. PMID- 6299203 TI - Two anatomoclinical types of warts with plantar localization: specific cytopathogenic effects of papillomavirus. Type I (HPV-1) and type 2 (HPV-2). AB - In this study, the clinical and histopathological aspects of 50 plantar warts are reported in relation to the type of papillomavirus present in the lesions, as detected by immunofluorescence tests, using specific guinea pig fluorescein labelled IgG. Warts of plantar localization are not caused by the same human papillomavirus (HPV) since they are found to be associated with both HPV type 1 (HPV-1) and HPV type 2 (HPV-2). HPV-1 is always associated with deep and painful plantar warts (myrmecia), whereas HPV-2 is found to be associated with superficial, painless plantar warts (vulgaris or often mosaic type). Histologically, these two types of plantar warts are quite different. In myrmecia (HPV-1), characterized by an endophytic growth, large eosinophilic, keratohyaline like granules are observed in the cytoplasm and nucleus of infected, often vacuolated cells. These granules appear early in stratum spinosum and are very numerous in stratum granulosum. In the mosaic type (HPV-2), the histopathological aspect is not different from that of common warts; these lesions have an exophytic growth and are characterized by foci of clear vacuolized cells which are found in stratum granulosum. Their cytoplasm contains round, basophilic keratohyalin granules which often have a heterogenous aspect. These differences are observed in other localizations of morphologically related warts associated with HPV-1 and HPV-2 and seem to be related to a specific cytopathogenic effect of HPV-1 an HPV-2 in human papillomas. PMID- 6299204 TI - Cutaneous cytodestructive potency of lignans. I. A comparative evaluation of influence on epidermal and dermal DNA synthesis and on dermal microcirculation in the hairless mouse. AB - The four lignans: podophyllotoxin, 4'-demethyl-podophyllotoxin, alpha-peltatin and beta-peltatin represent the podophyllin ingredients believed to exert lesional necrosis after application to condylomata acuminata. This study comparatively evaluates cutaneous cytodestructive potency of these drugs, as evaluated by measurement of epidermal and dermal 3H-thymidine incorporation and injury of dermal microcirculation at intervals in the range of 9--168 h after application to hairless mouse skin. Influence on DNA-synthesis was measured using a disc technique for isolation of standardized skin areas followed by alkali extraction of DNA from respective tissues after separation through potassium bromide incubation. Any capillary injury was demonstrated by visual assessment of leakage of i.v. injected trypan blue to test areas. An analytical method compensating for variations in DNA synthesis in individual controls is presented. Using 0.5% preparations of the lignans, podophyllotoxin most consistently influenced the variables indicating cytodestructive effect, while 4'-demethyl podophyllotoxin exerted the weakest degree of damage. PMID- 6299205 TI - Epidermodysplasia verruciformis induced by a new human papillomavirus (HPV-8). Report of a case without immune dysfunction. Effect of treatment with an aromatic retinoid. AB - A new case of epidermodysplasia verruciformis (EV) is reported in a 35-year-old man characterized by multiple common warts, flat warts, psoriasis-form lesions, and pityriasis-versicolor-(PV) like lesions. There was no familial history, no mental retardation, and no malignant changes. Human papilloma virus type 8 was identified in PV-like lesions. Immunologic studies detected no abnormalities: Delayed skin tests, DNCB sensitization, levels of immunoglobulins, complement components, circulating immune complexes, number of circulating lymphocytes, lymphocyte membrane markers, proliferative responses to mitogens, Ts activity, and Ts subpopulations as detected by monoclonal antibodies were considered to be within normal limits. A treatment with a retinoid aromatic (RO 10-9359) improved the clinical status of the patient and did not modify the immune parameters. PMID- 6299207 TI - Acceleration of the development of benzopyrene-induced skin cancer in mice by microwave radiation. AB - Development and growth of skin cancer may be affected by various physical and chemical factors present in human environment. Of these factors electromagnetic radiation of radio- and microwave spectra are among the most common. In the present study Balb/c mice were exposed to chemical carcinogen, 3,4-benzopyrene, painted on the skin every 2nd day for a total of 6 months, and simultaneously irradiated with athermal (5 mW/cm2) or subthermal (15 mW/cm2) doses of 2,450 MHz microwaves. The other group of animals was preirradiated with microwaves at 10 mW/cm2 power level for 1, 2, or 3 months and then treated with benzopyrene, as above. Control mice were exposed for 6 months to benzopyrene, resulting in the development of baso- or spinocellular skin carcinoma within approximately 9 months, and sham-irradiated with microwaves. The growth of the tumour was assessed according to a self-designed 7-range macroscopic scale, supported by microscopical examinations of skin sections. All protocols of microwave irradiations resulted in a significant acceleration of the development of benzopyrene-induced skin cancer and in shortening of life span of the tumour bearing hosts. This effect seemed to be dose-dependent since subthermal doses (15 mV/cm2) and longer (3 months) expositions to microwaves were more efficient as compared to athermal doses (5 mW/cm2) and shorter preirradiations. In addition, low-level, long-lasting exposure to microwaves led to a marked suppression of delayed hypersensitivity of mice treated with benzopyrene, as assessed by their reactivity to dinitrofluorbenzene (DNFB). It is suggested that the observed co carcinogenic effect of microwave radiation may, at least in part, result from the inhibitory action of microwaves on cellular immune reactions of exposed animals. PMID- 6299206 TI - Natural killer cell activity in atopic dermatitis. AB - Natural killer (NK) cell activity against K562 cells was studied in 12 male adults with atopic dermatitis (AD). In a 4-h chromium release microcytotoxicity assay normal NK cell function and its augmentation by interferon (IFN-alpha) were observed in AD patients. Slightly higher NK cell activity was observed in patients with allergic respiratory symptoms. The role of NK cells in the pathogenesis of atopic dermatitis is discussed. PMID- 6299208 TI - Susceptibility of Neisseria gonorrhoeae to cefotaxime: in vitro studies and treatment results. AB - More and more beta-lactamase-producing, penicillin-resistant strains of Neisseria gonorrhoeae are nowadays isolated in most parts of the world including the European continent. Therefore, there is a need for reliable alternatives in the treatment of gonorrhoea. Cefotaxime--first representative of a new generation of beta-lactamase-resistant cephalosporins--was evaluated for this reason, first in vitro and then in vivo. All 191 strains tested proved susceptible, the highest MIC amounting to 0.03 microgram/ml, a drug concentration well exceeded in plasma for more than 4 h after i.m. application of 0.5 g cefotaxime. In contrast to other investigators we have therefore decided to test this relatively small dose clinically without the addition of probenecid. The clinical results were excellent: 146 of 151 patients (96.7%) were cured. In the female sub-group (34 patients) the cure rate amounted to 100%. Thus, the i.m. injection of 0.5 g cefotaxime as a single dose must be looked upon as a reliable regimen for the cure of uncomplicated gonorrhoea, both in males and females. PMID- 6299209 TI - Accumulation of renal effects of lead in urban populations of feral pigeons, Columbia livia. PMID- 6299210 TI - Partitioning of polybrominated biphenyls (PBBs) in serum, adipose tissue, breast milk, placenta, cord blood, biliary fluid, and feces. AB - Polybrominated biphenyl (PBB) concentrations in specimens from adult males and females were used to determine the distribution of PBBs in body tissues and the partitioning ratio between types of tissues. Specimens of serum, adipose tissue, biliary fluid, and feces were tested by gas chromatography. In addition, parturient women provided breast milk, placenta, and cord blood for testing. There was a significant correlation between serum and adipose PBB levels. Pregnant and nonpregnant women and male chemical workers had similar serum to adipose tissue concentration ratios, which ranged from 1:140 to 1:260. Males from farms had a significantly different ratio of 1:325-329. Potential exposure to the fetus and newborn was demonstrated. Cord blood contained one-tenth of the concentration found in maternal serum which indicated partial placental passage. Human milk contained PBBs at 107 to 119 times the quantity found in maternal serum. Polybrominated biphenyls were detectable in bile and feces demonstrating transfer into the intestinal tract. The concentration of PBBs in feces represented a minor proportion of the total body burden indicating a slow rate of excretion. PMID- 6299211 TI - Late progression of radiographic changes in Canari chrysotile mine and mill exworkers. AB - The present radiological status of 133 exworkers (mean age: 58 +/- 0.7 yr) of the chrysotile mine of Canari (Corsica) was compared to that recorded in 1965, when the mine was closed. The mean time since first exposure is 27 +/- 0.4 yr, and the mean time since termination of asbestos exposure equals 16 +/- 0.3 yr. This study showed an evolution of radiographic changes: in 1965 only 14% of the subjects had grade 1/1 or more parenchymal fibrosis, and 6% had bilateral pleural lesions; in 1979 these percentages reached 40% and 27%, respectively. Subjects who had a given parenchymal change in 1965 (e.g., 1/1 grade lesions) were older and had a more intensive dust exposure and tobacco consumption than exminers who developed the same parenchymal changes much later. A progression of parenchymal changes was observed in 39% of the subjects with normal findings in 1965, and in 63% of the subjects with grade 1/1 or more changes in 1965. Those who progressed (46% of the cohort) were older, had a longer exposure, and a more intensive dust exposure and cigarette smoking than nonprogressors. PMID- 6299212 TI - Immunoperoxidase demonstration of human papilloma virus (HPV) in dysplastic lesions of the uterine cervix. AB - Routinely processed sections of 47 unselected cervical dysplasias and in situ carcinomas (both condylomatous, CO, and noncondylomatous, NCO) were subjected to staining with indirect immunoperoxidase-PAP technique using anti-human papilloma virus (anti-HPV) immune serum to demonstrate HPV antigens in these lesions. Anti HPV was raised in guinea pigs immunized with highly purified virions from a pool of human skin wart tissues. Positive reactivity for HPV (brown intranuclear precipitates) was disclosed in 72% of the CO dysplasias and in only one case (5.6%) of the NCO group. The frequency of HPV-positive lesions was inversely related to the degree of epithelial dysplasia, being consistent with the disappearance of ultrastructurally detectable virus particles in lesions turned malignant. HPV-containing cells were characteristic koilocytes or dyskeratotic superficial cells. The present technique provides definite evidence for the HPV etiology of the flat, inverted, and papillomatous condylomas, and seems to offer an advantage as a diagnostic tool of these lesions. When applied to malignant tumors in the near future, the method will undoubtedly bring more light on the suggested role of HPV in the etiology of human squamous cell carcinomas of the uterine cervix and other sites as well. PMID- 6299213 TI - [Mucous adenocarcinoma of the bladder]. PMID- 6299214 TI - Hypothyroidism and calcium pyrophosphate dihydrate deposition disease. PMID- 6299215 TI - Fate of herpes simplex virus in lymphocytes from blood and joint effusions of systemic and pauciarticular juvenile chronic arthritis. AB - The fate of herpes simplex virus (HSV) in the lymphocytes of patients with juvenile chronic arthritis (JCA) was studied by previously described methods. Lymphocytes isolated from blood and joint effusions of 18 patients with systemic JCA and 11 patients with pauciarticular JCA usually supported viral growth normally. PMID- 6299216 TI - Surgical implications of Klippel-Trenaunay syndrome. AB - Between January 1956 and July 1981, 40 patients with Klippel-Trenaunay syndrome were seen at the Mayo Clinic. Twenty male and 20 female patients presented with the classic triad of soft tissue and bony hypertrophy of the extremity, hemangioma, and varicosity without evidence of functional arteriovenous fistulae. The lower extremity was involved in 38 patients (95%), the upper extremity in six patients (15%). In four of these cases (10%), both the upper and lower extremities were affected. The disease was unilateral in 34 patients (85%), bilateral in five cases (12.5%), and crossed-bilateral in one case (2.5%). Surgery was done in 13 patients (32.5%), nine of whom were operated on at the Mayo Clinic and four of whom had had previous surgery elsewhere. Excision and stripping of varices were performed in three patients; of these three, a deterioration of symptoms was observed in one, but the procedure was beneficial in the other two. Partial varicectomy was performed in four cases, while resection of the angioma was attempted in eight cases, with good results in three cases of small angiomas. Femoral and tibial epiphysiodesis stopped the overgrowth and produced an excellent result in one case. In four cases of epiphysiodesis at the foot level and two cases of derotational tibial osteotomy, moderate improvement was achieved. The indication for vascular or orthopedic surgery should be carefully considered in each patient who has this syndrome. One patient (2.5%) died of a severe form of the disease, but the relatively benign course is documented by the 21 patients (52.5%) who are free of complaints without any treatment or with elastic support only. PMID- 6299218 TI - Interaction of the benzodiazepine antagonists, CGS 8216 and Ro 15-1788, with central adenosine A-1 receptors. AB - Although the potent benzodiazepine antagonist CGS 8216 has in vitro activity indicative of an interaction with central adenosine systems, neither it, nor the benzodiazepine antagonist Ro 15-1788 showed any great degree of interaction with central adenosine A-1 receptors as measured by radioligand binding. It is concluded that interaction of benzodiazepine antagonists with A-1 receptors is not a property related to their antagonist activity. PMID- 6299217 TI - Major hepatic resection. A 25-year experience. AB - Major hepatic resections were performed on 138 patients for a variety of conditions. There was one intraoperative death. Including this patient, there were 15 deaths within 30 days of the operation (operative mortality 10.9%). Important postoperative complications were intra-abdominal sepsis (17%), biliary leak (11%), hepatic failure (8%), and hemorrhage (6%). The results of 30 resections for the benign lesions, liver cell adenoma, focal nodular hyperplasia, hemangioma, and cystadenoma showed no operative mortality and low morbidity. Of 26 patients with hepatocellular carcinoma, seven died within a month of operation. The cumulative survival of the 26 at five years was 38%, and of the 19 who survived the procedure, 51%. Poor survival followed resections for cholangiocarcinoma and "mixed tumors." The five-year cumulative survival of 22 patients who had colorectal metastases excised was 31%. Apart from a patient with carcinoid, prolonged survival was rare after resection of other secondaries and after en bloc resections for tumors directly invading the liver. Hepatic resection was of value in the management of some patients with hepatic trauma, Caroli's disease, liver cysts, and intrahepatic stones. PMID- 6299219 TI - Pre-synaptic and post-synaptic effects of xylazine and naphazoline on the bisected rat vas deferens. AB - The effect of the selective alpha 2-adrenoceptor agonists, naphazoline and xylazine, was studied on the field stimulated bisected rat vas deferens. Xylazine inhibited the twitch response to field stimulation in both the prostatic (ID50 = 0.10 microM) and epididymal (ID50 = 0.08 microM) halves of the rat vas deferens. This effect was antagonized by yohimbine (0.01-0.10 microM). Naphazoline also inhibited the response to field stimulation in the prostatic (ID50 = 0.12 microM) but had no such action on the epididymal half of the rat vas deferens. Indeed, low concentrations of naphazoline (threshold, 0.05 microM) contracted the epididymal vas deferens preparation. These contractions were competitively antagonized by prazosin suggesting an action on post-synaptic alpha 1 adrenoceptors. Neither pre-synaptic nor post-synaptic actions of either drug were affected by cocaine (10 microM) or beta-oestradiol (10 microM) added to the Krebs' solution. The results provide further evidence for the existence of two types of post-synaptic alpha 1-adrenoceptors and suggest a different anatomical localization of these receptors between the two ends of the rat vas deferens. PMID- 6299220 TI - The inhibitory effect of newer calcium antagonists (nimodipine and PY-108-068) on vasoconstriction in vivo mediated by postsynaptic alpha 2-adrenoceptors. AB - In the pithed rat, stimulation of postsynaptic alpha 1 as well as alpha 2 adrenoceptors is known to cause vasoconstriction. The two newer nifedipine related calcium antagonistic agents nimodipine and PY-108-068 proved potent inhibitors of pressor responses induced by the selective alpha 2-receptor agonist B-HT 920. The maximal pressor effect of B-HT 920 was significantly reduced, whereas the hypertensive effects brought about by selective postsynaptic alpha 1 adrenoceptor stimulation were, but slightly, impaired by nimodipine and PY-108 068 and the maximal increase in diastolic pressure was not affected by the calcium antagonists. Under beta-blockade and suppression of alpha 1-adrenoceptor mediated vasoconstrictor effects by prazosin, the resulting alpha 2-adrenoceptor induced vasopressor response to (-)-norepinephrine proved also very sensitive towards the inhibitory action of nimodipine and PY-108-068. In comparison with nifedipine, PY-108-068 was 3.9 and nimodipine 5.1-times more potent with respect to reducing vasopressor responses of B-HT 920. PMID- 6299221 TI - [Effects of the analog ACTH 4-10-7-L phenylalanine on cerebral protein metabolism]. PMID- 6299222 TI - [Adrenoleukodystrophy in children. Apropos of 20 cases]. AB - Adrenoleukodystrophy (ALD) is a sex-linked recessive metabolic encephalopathy usually beginning in childhood. This series of 20 cases showed the following associated symptoms: mental deterioration (20 cases), behavior modification (19 cases), decreased visual acuity (17 cases), deafness (13 cases) and seizures (7 cases). Adrenal insufficiency often consisted only of melanoderma (13 cases). Diagnosis was assessed by characteristic CT-scan findings (8 of 13 cases), adrenocortical insufficiency partial glucocorticoid (19 of 19 investigated cases) or global, mineralo- and involving both aldosterone and glucocorticoid hormones (9 of 17 investigated cases) and by the presence of specific inclusions on the skin and/or conjunctival biopsy (5 of 7 cases). PMID- 6299223 TI - [Distribution of lung cancer metastases. II. Effect of tumor histology]. AB - The various histological types of 1,283 lung cancers show remarkable differences in sex distribution, in mean age at the time of death, in the localization of tumors and duration of cancer disease as well as in their metastatic behaviour. The proportion of the various histological types of lung cancer in ten organs with blood borne metastases has been analysed. Metastases of small cell carcinomas most frequently occur in pancreas, liver, thyroid gland, bone and adrenal, whereas squamous cell carcinomas are found in kidney and heart. Anepidermic squamous cell carcinomas are most frequently seen in brain and large cell carcinomas in the spleen. As already mentioned in the preceding paper, combinations of metastatic affected organs can be seen with special frequencies in all histological types of lung cancer. However, small cell carcinomas provide the most evident findings. PMID- 6299224 TI - The light and electron microscopic localization of intracytoplasmic nucleolus like bodies in the mouse brain stained by Holmes' silver method. AB - By light microscopic observation of the mouse brain impregnated by the Holmes' method, intensely stained cytoplasmic inclusion bodies were demonstrated in the nerve cells of the hypothalamus, locus coeruleus and other autonomic regions in addition to the limbic system and some circumventricular organs. Electron microscope observation of ultrathin sections made from vibratome sections impregnated by the Holmes' method revealed that the structures which have been called the nucleolus-like bodies were loaded with large amounts of gold particles. Hence, the Holmes-impregnated cytoplasmic inclusions could be identified with the so-called nucleolus-like bodies. The fine structure, distribution and significance of these bodies were described. PMID- 6299225 TI - The fine structure of the follicle-forming cells in the adenohypophysis of the ironfish, a natural hybrid between the Funa and the goldfish (Teleostei, Cyprinidae). AB - The ironfish considered to be the natural hybrid between the Funa (Carassius carassius) and goldfish (C. auratus), has several peculiarities in its morphology, karyology and reproductive biology. Its adenohypophysis also shows follicular formations which are never found in either parent species. The follicles of the ironfish adenohypophysis consist of both prolactin cells and stellate (agranular) cells. The prolactin cells have a tall pyramidal form, and their secretory granules are seen more often in the basal cytoplasm than in the apical region. The electron-dense content of the granule is released into the perifollicular space by exocytosis. The apices of the cells bearing microvilli and a cilium are exposed to the follicular lumen. Many of the cilia, however, protrude from the lateral surface of the apical region into the intercellular cleft. The apical processes of the stellate cells extend among the prolactin cells and reach the follicular lumen. The end of the process becomes broad with microvilli and a cilium, and is joined together with the apical processes of other stellate or prolactin cells and sometimes lies in close proximity to the corticotrophs near the follicle. Nerve terminals in the perifollicular region contain small clear vesicles and large granular vesicles. They are occasionally in direct contact with prolactin cells and corticotrophs. By fluorescence histochemistry for catecholamines, blue-green fluorescent fibers are frequently detected in the RPD but rarely near the follicle. These findings suggest that the follicles seen in the ironfish are derived from the regional rudiments of the Rathke's pouch. PMID- 6299226 TI - Scanning electron microscopic observations of rat tibia using the HCl-collagenase method. AB - The metaphysis of rat tibia was examined by scanning electron microscopy (SEM) using the HCl-collagenase method (EVAN et al., 1976) and the three-dimensional structure of osteocytes and the relationship between osteocytes and osteoblasts were demonstrated. After the HCl-collagenase treatment, the bone matrix near the exposed surface was completely removed but the cellular components retained their structural integrity. Young osteocytes located close to osteoblasts showed flattened, spindly shapes. They were connected with adjacent osteoblasts by short, wide and slightly branched processes. On the other hand, the osteocytes located in the deeper portion of trabecular bone were smaller in size, being round or oval and flattened in shape. They were connected with both osteoblasts and neighboring osteocytes by long and more extensively branched processes forming a complicated network. From these results, it is suggested that osteoblasts extend their processes perpendicularly to the bone surface, with these processes branching off and connecting to each other at the osteoid layer. PMID- 6299227 TI - Dense-core matrical mitochondrial bodies in oncocytic adenoma of the thyroid. AB - Single and/or multiple, round and homogeneous electron-dense bodies occurred within the mitochondrial matrix of a mitochondrion-rich follicular adenoma of the thyroid. These non-membrane-bound bodies varied greatly in size and often occupied a large portion of the mitochondrion. They were present in normal-sized or large mitochondria that exhibited cristolysis and/or dilatation of the intracristal space of the residual cristae and formation of intramatrical myelin figures. The globule-containing mitochondria frequently exhibited bundles of parallel intramatrical filamentous inclusions that were 6 to 8 nm in width and were randomly arranged in clusters. Megamitochondria up to 4.4 micrometers in widest diameter occurred in a frequency of one to two per cell and they displayed similar bundles of filamentous inclusions. This evidence of electron-dense bodies in non-steroid-hormone-secreting cells suggests that the bodies result from nonspecific mitochondrial reaction to injury rather than being inclusions specific for steroid-synthesizing cells. PMID- 6299228 TI - Pitfalls in tumours of the knee. AB - The authors describe two patients with knee symptoms. In both cases the correct diagnosis was established late. One patient had a large cyst of the lateral meniscus, while the other had a synovial sarcoma. Aetiology, symptomatology, methods of investigation, therapy and prognosis are discussed, as well as the differential diagnosis. It is stressed, that a meniscal cyst is not adequately treated until the entire meniscus is removed. The puzzling signs and symptoms of a synovial sarcoma of the knee are mentioned. It is obligatory to perform a biopsy in all patients with an unfamiliar swelling of the knee, in order to rule out a malignancy, especially the treacherous synovial sarcoma. PMID- 6299230 TI - Lymphedema treated with a linear pump: pediatric case report. AB - This first report on the use of the Wright linear pump and its successful application in the treatment of a child with Park-Weber syndrome indicates that sequenced linear pumping is efficacious when previously tried single cell intermittent compression fails. Programmable intermittent pressure in the management of lymphedema in children offers a preferable choice. PMID- 6299229 TI - Axillary F central latency: simple electrodiagnostic technique for proximal neuropathy. AB - A simple technique is described for measuring the F-wave conduction time of the proximal segment of peripheral nerves. This should prove helpful in the diagnosis of a proximal neuropathy or myelopathy. The F wave (Fw) and M response at the wrist (Mw) and the M response at the axilla (Ma) can be used to calculate the conduction time, defined as Axillary F Central Latency (AFCL), between the spinal cord and the axilla, covering a horizontal distance of 25cm twice. The calculated AFCL (AFCL = (Mw + Fw) - 2Ma) in our control group of 30 normal subjects was 11.2 +/- 0.8 msec (mean and 1 standard deviation) for the ulnar or median nerve. An AFCL of more than 13.6 msec (mean plus 3SD), or a discrepancy of the AFCL greater than 2.1 msec in the same nerves between the 2 sides or between the median and ulnar nerves on the same side suggests an abnormality involving the proximal nerve segment. The prolonged AFCL has been found in patients with Guillain-Barre syndrome (GBS), brachial plexus lesions, syringomyelia and, in a few cases, with thoracic outlet syndrome (TOS). PMID- 6299231 TI - Susceptibility of Marek's disease lymphoblastoid cell lines to infection with influenza and pseudorabies viruses and the protective effect of immunization with influenza virus-infected lymphoblastoid cells. AB - The susceptibility of 3 lymphoblastoid cell lines (MDCC-MSB1, MDCC-HP1 and MDCC RP1) derived from Marek's disease tumours to infection with influenza (AoPR8) and pseudorabies viruses was studied. MSB1 and HP1 were more susceptible to infection with influenza virus than was RP1. It was shown in the case of HP1, which was studied in greater detail, that although the majority of the cells synthesized influenza virus haemagglutinin when infected at high multiplicity, only a small proportion produced mature infectious particles. The results obtained from both high and low multiplicities of infection suggested that infection with AoPRs was abortive in the majority of the cells of all 3 cell lines. Infected cultures remained viable when subcultured over several days. All 3 cell lines were resistant to pseudorabies virus. The block in pseudorabies virus replication probably occurred after adsorption. Immunization of chickens with HP1 lymphoblastoid cells superinfected with influenza virus AoPR8 and inactivated with glutaraldehyde gave better protection against Marek's disease than immunization with uninfected HP1 cells. PMID- 6299232 TI - Characterization of an herpes simplex virus type 2 mutant, which is resistant to acycloguanosine and causes fusion of BSC1 cells. AB - A mutant of herpes simplex virus type 2, which induces low levels of thymidine kinase activity in infected BSC1 cells and consequently able to grow in the presence of acycloguanosine, was isolated. This mutant has also been shown to cause fusion of BSC1 cells. In BSC1 cells, co-infected with the wild-type strain and the mutant, the yield of each of the two viruses was normal but the rounding and aggregation of cells observed, resembled that found in wild-type infected cultures. When the mixed infection was performed in the presence of acycloguanosine (100 micrometers), the growth of the two virus strains was inhibited, as well as the cytopathic effect in the cultures. It is suggested that under these conditions, the thymidine-kinase which was induced in the infected cells by the wild-type strain, phosphorylated acycloguanosine and the activated drug formed, inhibited the growth of the two viruses by interference in their DNA syntheses. PMID- 6299233 TI - Ultrastructural localization of viral antigen in nuclear inclusions of cytomegalovirus infected guinea pig cells. AB - Intranuclear localization of viral antigens in guinea pig cytomegalovirus (GPCMV) infected guinea pig embryo (GPE) cells was investigated by cross-reactive indirect immunoperoxidase and immunoferritin techniques utilizing guinea pig antisera to GPCMV. Following primary fixation with 4 percent paraformaldehyde, a brief treatment of infected cells with 0.25 percent trypsin was found to enhance penetration of antibodies and the conjugates. Ferritin or horseradish peroxidase conjugated goat anti-rabbit IgG was used as a secondary antibody that cross reacted with guinea pig immunoglobulins in order to reduce non-specific immunochemical reactions. Using light microscopy following immunoperoxidase staining, GPCMV antigens in an intranuclear location were not discernable when the infected cells were stained without pretreatment with trypsin, however intranuclear GPCMV antigens could be visualized after the fixed cells were treated with trypsin for 2-4 minutes prior to addition of the antiserum. Electron microscopic examination following indirect immunoferritin staining revealed viral antigens localized on viral capsids and on scattered electrondense amorphous matrices but not on the surrounding tubular structures on fibrils. The possibility that tubular structures may be a host cell product produced in response to GPCMV infection is discussed. PMID- 6299234 TI - Cell-mediated immunity in mice with primary, secondary and latent herpes simplex virus infection. AB - Cell mediated immunity was studied by a cytopathic effect inhibition assay in mice infected in the ear with herpes simplex virus type 1 (HSV 1). Activity appeared rapidly, reaching a high level 6 days after primary infection. It had fallen 10 days after infection and was undetectable during latency, 3-5 weeks after infection. The activity reappeared even more rapidly and strongly after reinoculation with the virus, but stimuli designed to induce recurrent disease did not induce clinical disease in the animals and no activity was detected in them. The activity, which was specific for HSV, was shown to be mediated by T lymphocytes. PMID- 6299235 TI - Imide derivatives of 3-nitro-1.8-naphthalic acid: their inhibitory activity against DNA viruses. AB - The antiviral action of a new drug, 5-amino-2-(dimethylaminoethyl)-benzo-[de] isoquinolin-1.3-dione has been studied against herpes simplex type 2 (HSV-2) and adenovirus type 5 (Ad-5) grown in Vero cells. The concentration of the drug which gives a 5 log10 reduction in virus titer was 4 micrograms/ml (maximum tolerated concentration) for HSV-2. The anti-HSV-2 activity of this compound was one log. more powerful than that of iododeoxyuridine (IDU). At this concentration the drug shows virucidal activity against HSV-2. No inhibition was found when the drug was tested against Ad-5. The inhibition of virus production has been studied depending upon the amount of drug or virus, and drug addition-time after infection. Reversibility of inhibition after drug removal, and drug resistance in the presence of the drug have also been determined. PMID- 6299236 TI - Effect of acyclovir and presence of cellular and viral thymidine kinase activity in herpes simplex virus infected cells. Brief report. AB - Herpes simplex type 1 (HSV-1) infected human lung fibroblasts show a higher viral thymidine kinase activity than infected African green monkey kidney cells. This may partly account for the higher antiviral activity of acyclovir in the former cells. PMID- 6299237 TI - The production of varicella Zoster virus antiserum in laboratory animals. Brief report. AB - Hyperimmune anti-varicella zoster virus (VZV) antisera were prepared in BALB/c mice, Lewis rats and New Zealand rabbits by subcutaneous inoculations of purified VZ virions suspended in Freund's adjuvant. VZV antibodies were demonstrated both by an enzyme-linked immunosorbent assay (ELISA) and by indirect immunofluorescence (IF). VZV could not be detected in organs of any inoculated animals by IF or by cocultivation of tissue fragments from infected animals with human diploid fibroblasts. PMID- 6299238 TI - Orbi- and bunyaviruses from a puffin colony in the Outer Hebrides. PMID- 6299239 TI - Abortive infection of neural cells by herpes simplex virus type 2. AB - The nature of the refractoriness of C6 rat glioma cells to herpes simplex virus type 2 (HSV-2) was examined. Infection of C6 cells with HSV-2 results in low virus yields, not exceeding the input virus. Although virus growth studies suggested a restricted cycle of virus replication, synthesis of HSV-2 DNA and HSV 2-specific antigens could not be detected. In addition, HSV-2 yields in C6 cells were unaffected by interferon, cycloheximide, tunicamycin, actinomycin D and cytosine arabinoside. However, trypsin, but not EDTA, treatment of infected C6 cells at 4 hours postinfection (p.i.) reduced maximal HSV-2 yields at 24 hours p.i. by 61 percent. These data: 1) indicate that HSV-2 fails to replicate in C6 cells and is prohibited from directing the synthesis of virus macromolecules; and 2) suggests that the increment of HSV-2 yields observed during the synthesis phase of the virus growth cycle represents re-envelopment and egress of a portion of the input virus. PMID- 6299240 TI - Virus-specific sequences within restriction DNA fragments of cells persistently infected with tick-born encephalitis virus. PMID- 6299241 TI - Detection of cellular receptors for Sendai virus in mouse tissue sections. AB - We have established a method for detecting virus receptors in the tissue sections and have been able to clarify the distribution of cellular receptors for Sendai virus in various mouse organs, including trachea, esophagus, large intestine and cerebrum. The virus receptors were detected in ciliated cells of the trachea, as to be expected for a pneumotropic virus. The lamina propria and connective tissue of the mouse esophagus and large intestine, immediately under the epithelium, contained an abundance at virus receptors, but very few were detected in the epithelium of these organs. In mouse cerebrum, the cells lining the ventricles had a large number of virus receptors. PMID- 6299242 TI - Antibody to human leucocyte interferon may enhance the frequency of rhinovirus isolation. PMID- 6299243 TI - Baboon Endogenous C type Virus induces syncytium formation in Epstein barr virus (EBV) carrying lymphoblastoid human cell lines. PMID- 6299244 TI - Mink type C virus: biochemical characterization of the structural polypeptides. AB - Mink type C virions contained six major protein species of approximate M.W. of 90,000, 70,000, 30,000, 15,000, 12,000 and 10,000. The two largest polypeptides were glycosylated and the 12,000 M.W. polypeptide was the major phosphoprotein of the virion. Two-dimensional tryptic peptide map of the 30,000 M.W. major structural protein of MiLV showed a pattern distinct from those of analogous proteins from mouse and endogenous cat type C viruses. Significant peptide homology of this protein was, however, found with the corresponding protein of infectious feline type C virus (FeLV). PMID- 6299245 TI - Occurrence and reactivation of latent Aujeszky's disease virus following challenge in previously vaccinated pigs. AB - Pigs which were vaccinated with an inactivated ADV vaccine developed a latent ADV infection up to 18 months after ADV challenge. Up to 6.5 months p.i. latent virus could be demonstrated by co-cultivation of different organ tissues (lungs, tonsils, olfactory bulb, brain stem, medulla). Afterwards, reactivation of latent virus was only achieved by immunosuppression of the animals. Immunosuppression led to a limited virus replication in nasal mucosa, tonsils, lymph nodes and central nervous system. In addition ADV was detected in the nasal secretions., Humoral and cellular immunity was investigated before and after immunosuppression of the animals. Before immunosuppression most of the animals displayed SCC, ADV ADCC and ADV-LYST, and all animals had medium to high titres of neutralizing serum antibodies. After immunosuppression the number of pigs reacting in SCC and ADV-LYST, assays was distinctly reduced, but the number of animals reacting in ADV-ADCC assays remained unaltered. A significant reduction of serum antibody titres occurred only in 2 of 12 animals one day after the end of immunosuppression. PMID- 6299246 TI - A biochemical and biological comparison of three European isolates of nuclear polyhedrosis viruses from Agrotis segetum. AB - Isolates of multiply-enveloped nuclear polyhedrosis viruses from Agrotis segetum (Lepidoptera: Noctuidae) populations in England (AsNPVE), France (AsNPVF) and Poland (AsNPVP), were compared biochemically and for their infectivity to A. segetum and Mamestra brassicae larvae. The electrophoretic profiles of DNA restriction endonuclease fragments and viral proteins appeared identical for AsNPVE and AsNPVF. AsNPVP was distinct by these techniques, although some of the virus particle polypeptides had the same mobilities as those of the other isolates. A serological comparison of the A. segetum NPV isolates with other baculoviruses, using indirect enzyme-linked immunosorbent assay, suggested that AsNPVP was no more closely related than M. brassicae NPV to the other A. segetum NPV isolates. AsNPVP had significantly lower infectivity for neonate A. segetum larvae (LD50 = 350 inclusion bodies) than AsNPVE (10 inclusion bodies) or AsNPVF (23 inclusion bodies). AsNPVE and AsNPVF did not appear to replicate in M. brassicae larvae, while AsNPVP produced a limited infection in this species. PMID- 6299247 TI - Study on West Nile virus persistence in monkeys. AB - Experiments in M. rhesus showed persistence to be a typical property of West Nile virus. This property was exhibited by strains belonging to different antigenic types, and varying in virulence and in the isolation area (U.S.S.R., Uganda, India). The duration of persistence was at least 5 1/2 months in asymptomatic infection and in convalescence after encephalitis or a febrile disease. The virus isolated within the first 2 weeks after inoculation of monkeys has the standard properties. The virus persisting for 2 months retains its cytopathic and antigenic activity, however, is non-pathogenic for white mice. After 5 1/2 months of persistence the virus has no neurovirulence or cytopathic properties but is capable of infecting the susceptible cells and induces in them the synthesis of virus-specific antigen detectable by immunofluorescence. The persisting virus has been isolated by cocultivation of trypsinized monkey organ cells and cells of the indicator culture. This virus was located mostly in the cerebellum, cerebral subcortical ganglia, lymph nodes, and kidneys. The monkeys experiencing encephalitis, febrile, or asymptomatic infection showed in morphological examinations a subacute inflammatory-degenerative process in the central nervous system. The results suggest that West Nile virus, one of the most widely spread arboviruses in Africa, Asia, and Europe, may be implicated in the etiology of subacute diseases of the CNS. PMID- 6299248 TI - [Manifestations of the neurotoxicity of the organochlorine pesticide dilor during the postnatal period in the rat]. AB - When pregnant rats are administered chlororganic pesticide dilor, certain ultrastructural changes are observed in neurocytes and in myelin fibers, in the spinal cord, that demonstrates some disorders in cellular and tissue metabolism. In the newborn spinal cords a retarded differentiation is observed against the background of an intense cellular metabolism. By means of the electron paramagnetic resonance technique, an increasing concentration of free radicals in the brains and spinal cords of the pregnant animals and in their one-month-old offsprings is demonstrated. The investigation on fatty acid composition of lipids in pregnant test animals demonstrates a decrease in cholysterine, steroids and some fatty acids in myelin fractions and in synaptosomic membranes. PMID- 6299249 TI - [Immunocytochemical study of the myoepithelial cells in fibrocystic disease and ductal carcinoma of the breast]. AB - Myoepithelial cells (MC) were identified and the pattern of their distribution in dysplasia and duct carcinoma of the mammary gland was studied using a monospecific antiserum to smooth-muscle myosin of the human uterus, by the indirect Coons' method under luminescent microscope. Samples of the operation material from 10 patients with fibrous-cystic mastopathy and 26 patients with duct carcinoma were examined. In solid (7 observations) and comedo (4 observations) carcinomas with the structure of cancer in situ, MC were found intact in the periphery. In invasive growth of these tumors as well as in scirrhus (5 observations) and adenocarcinoma (10 observations) MC disappeared from the duct structures and were absent in carcinoma proliferates. Positive immunofluorescence in the conventional location of MC in the alveolar duct system of the mammary gland allows sufficiently valid identification and study of the distribution and changes in these cells. The immunocytochemical verification of MC in dysplasia and duct carcinoma indicates the possibility of using MC as markers of duct carcinoma in situ; MC disappearance from the duct structures characterizes the beginning of the infiltrative tumor growth. PMID- 6299250 TI - [Multicentric retroperitoneal malignant fibrous histiocytoma]. PMID- 6299251 TI - [Multiple mixed adenoma of the liver associated with small-cell carcinoma and pulmonary tuberculosis with cystadenopapilloma of the kidneys]. AB - Cholangio- and hepatocellular adenomas of the liver occur very rarely. An observation of a man of 55 is presented who, along with the above pathology, had in the liver areas of a cartilagelike tissue with cavities filled with erythrocytes and resembling bone marrow canals which emphasized the dysembryogenetic origin of the tumor. No description of such combined pathology could be found in the available literature. PMID- 6299252 TI - Advantages of bicarbonate hemodialysis. AB - Six patients with frequent episodes of symptomatic hypotension during acetate dialysis were treated with bicarbonate dialysis. In all patients blood pressure, heart rate, and arterial acid-base values were measured every 30 minutes during each of the five treatments with acetate dialysis and bicarbonate dialysis. Hemodynamic parameters were measured invasively in all patients during bicarbonate dialysis and in three of them also during acetate dialysis. Additionally, continuous long-time monitoring with electroencephalography was performed during acetate dialysis and bicarbonate dialysis. During acetate dialysis the patients showed a frequent onset of sudden hypotension and arrhythmia with concomitant symptoms of the so-called disequilibrium syndrome, whereas these symptoms were nonexistent in the same patients during bicarbonate dialysis. PMID- 6299254 TI - Short- and long-latency median somatosensory evoked potentials. Findings in patients with localized neurological lesions. AB - Short- and long-latency somatosensory evoked potentials (SEPs) were elicited by stimulation of the median nerve in 43 patients with neurological disorders. Abnormalities of short-latency peaks, P9, N13, and P14, were seen in patients with lesions of the peripheral nerve, cervical spinal cord, and brain stem, respectively. Subsequent component, N18, was affected in patients with thalamic or hemispheric disease. In some patients with parietal lobe lesions, however, abnormalities were limited to later components, N32 or N63. Analysis of SEPs is helpful in localizing a lesion along the somatosensory pathway, although differentiation between thalamic and other subcortical or cortical involvement may not be possible with the present SEP technique. Both short- and long-latency SEPs should be studied for maximal clinical information. The latter can be most reliably evaluated by simultaneous bilateral stimulation. PMID- 6299253 TI - Clinical experience of postoperative hepatic failure treatment with pig or baboon liver cross-hemodialysis with an interposed membrane. AB - The present study emphasizes the principle of using liver support to restore the blood ketone body ratio (acetoacetate/beta-hydroxybutyrate), which reflects the redox potential of liver mitochondria and correlates with hepatic energy charge (ATP + 0.5ADP/ATP + ADP + AMP). Eleven surgical patients with grade IV hepatic coma were treated by an ex vivo pig or baboon liver cross-hemodialysis with an interposed Cuprophan membrane when their blood ketone body ratios had decreased to below 0.4 compared with the normal of above 0.7. Three patients were treated by cross-hemodialysis using a standard Cuprophan membrane dialyzer without increase of blood ketone body ratio and without marked beneficial effect. However, five of eight patients who had blood ketone body ratios of above 0.25 became fully alert after treatment by cross-hemodialysis using the larger pore size and greater surface area Cuprophan membrane, concurrent with a rise in the decreased blood ketone body ratio, and three of them were later discharged. By contrast, in the three patients with blood ketone body ratios below 0.25, there was no restoration of consciousness and no improvement in their blood ketone body ratios by this liver support. It is suggested that, as long as the blood ketone body ratio remained over 0.25, this metabolic liver support is effective in restoring grade IV hepatic coma. PMID- 6299255 TI - Clinical and histologic findings in opportunistic ocular infections. Part of a new syndrome of acquired immunodeficiency. AB - White retinal opacities and ocular inflammation may be two of the early signs of a new syndrome of acquired immunodeficiency. This syndrome primarily affects young homosexual men and is characterized by acquired cellular immunodeficiency in infections with multiple opportunistic microorganisms in addition to Kaposi's sarcoma. The apparently innocent ocular findings may be the first harbingers of this devastating syndrome. PMID- 6299256 TI - Fibrous histiocytoma of the corneosclera. AB - A 65-year-old woman had a slowly progressive sclerocorneal lesion that, after three years of unsuccessful medical treatment, involved almost the whole cornea and the surrounding sclera in the temporal lower quadrant. The tumor had a nodular surface. During the next two years, the tumor was operated on four times (twice by keratectomy and twice by lamellar sclerokeratoplasty). Microscopic examination revealed a fibrous histiocytoma. The resection was histologically incomplete on all occasions. Five years after the onset of the disease, the eye was enucleated. A spontaneous invasion of the iris could not be diagnosed with certainty, because there was a perforation at the limbus during a preceding operation. PMID- 6299257 TI - Severe anemia as a manifestation of metastatic jugular paraganglioma. AB - A young woman who had a right glomus jugulare paraganglioma had diffuse pulmonary metastases three years after surgical excision of the paraganglioma. Associated with these developments were profound anemia and an extraordinarily rapid ESR. These findings have been previously noted in patients with metastatic paraganglioma and have not as yet, to our knowledge, had a satisfactory explanation. Noteworthy in our patient was a diminished serum erythropoietin level, which may indicate that metastatic paragangliomas inhibit production of or interfere with maintenance of serum erythropoietin. Symptomatic palliation of the severe anemia was attained in this patient by injections of nandrolone decanoate. Follow-up examinations of patients with paragangliomas should include surveillance of the CBCs and ESR, both of which may reflect tumor activity. PMID- 6299258 TI - Viruses recovered from mosquitoes and wildlife serum collected in the Murray Valley of South-eastern Australia, February 1974, during an epidemic of encephalitis. AB - Pools of mosquitoes collected in the Murray Valley in February, 1974, during an encephalitis epidemic yielded 239 isolates of 11 distinct viruses. These included 39 isolates of MVE virus, an incriminated causative agent of encephalitis in man, and 111 isolates of Kunjin virus, a probable causative agent. An additional isolate of MVE virus was recovered from the serum of a white-faced heron, Ardea novaehollandiae. The other 9 viruses comprised the Alpha-viruses Ross River and Sindbis, the Flavivirus, Edge Hill, the Bunyaviruses, Koongol and Wongal, the undefined Kowanyama virus, and 3 previously undescribed viruses. One isolate of MVE virus was recovered from a total of 180 Culex australicus, and all other isolates of all viruses were from Culex annulirostris. The combined apparent infection rate of MVE and Kunjin viruses in Cx annulirostris was 1/913. The concurrent activity of so many seemingly transient arboviruses, their wide geographical distribution and temporal considerations all weaken the widely held hypothesis that during the pre-epidemic period MVE virus is introduced into the Murray Valley by viraemic waterbirds from distant areas of tropical enzooticity. PMID- 6299259 TI - Variation in arbovirus infection rates in species of birds sampled in a serological survey during an encephalitis epidemic in the Murray Valley of South eastern Australia, February 1974. AB - There was extensive and exuberant breeding of waterbirds before and during an epidemic of arboviral encephalitis in the Murray Valley of south eastern Australia in 1974. As estimated by haemagglutination inhibition tests on 432 bird sera collected between 4th and 13th February, 1974, infection with Murray Valley encephalitis virus, Kunjin virus and possibly other flaviviruses was concentrated in species of the Order Ciconiiformes (55% positive) and Pelecaniformes (41%), compared with only 5% in Anseriformes. Although Sindbis virus infections were also highest in these 2 Orders (56% and 46%, respectively), the incidence of antibodies was spread more uniformly through other Orders than with the flaviviruses: e.g. Anseriformes, 33%, Podicipitiformes, 27%. As viruses were recovered virtually only from Culex annulirostris mosquitoes, the different patterns of infection seem unrelated to host preference by mosquito species or behavioural response to mosquito attack, and suggest a specific relationship between MVE/KUN and Ciconiiformes and Pelecaniformes. The highest infection rate was 22/25 in mature rufous night herons (formerly nankeen night herons) Nycticorax calendonicus, and here tolerance to mosquito attack was probably a contributing factor. Assays of sera from 13 horses indicated high infection rates both alphaviruses and flaviviruses. PMID- 6299260 TI - Aspiration of breast cysts. AB - One hundred and sixty nine women with 220 breast cysts were treated by needle aspiration between 1950 and 1980. Mean follow up was seven years. One hundred and twenty two (72%) patients were aged from 40 to 49 years. The majority (61%) of cysts occurred in the left breast. In 147 patients 174 (79%) of 220 cysts were aspirated once and did not recur at that site. One hundred and eleven (66%) patients presented with a single cyst and had no recurrence in either breast after aspiration. Two patients developed carcinoma of the opposite breast at 8 and 12 years after cyst aspiration. Needle aspiration of the breast is regarded as a safe, simple and economic diagnostic and therapeutic modality in women presenting with single breast lumps. PMID- 6299261 TI - Breast localization biopsy. AB - Nineteen localization biopsies were performed on eighteen women, where routine mammography revealed impalpable abnormalities. These abnormalities are described and the results of biopsy given. Six carcinomas were detected, all of which were shown to have no evidence of metastatic disease. PMID- 6299262 TI - Cytology: a useful adjunct in the management of nipple discharge. PMID- 6299263 TI - Beta-endorphin levels in newborn cerebrospinal fluid. PMID- 6299264 TI - Additivity of neurochemical changes in learned helplessness and imipramine. PMID- 6299265 TI - Naloxone enhancement of memory processes: effects of other opiate antagonists. PMID- 6299266 TI - Transferred odor aversions in adult rats. PMID- 6299267 TI - Deoxythymidine kinases in varicella-zoster virus infected and biochemically transformed cells. AB - Deoxythymidine kinase (TK) activity induced in varicella-zoster virus (VZV) infected human embryonic fibroblast (HEF) cells was immunologically distinguishable from that in non infected HEF cells and also from that in herpes simplex virus type 1 (HSV-1) infected HEF cells. The TKs in VZV-biochemically transformed cells were immunologically the same as that induced in VZV-infected human cells and immunologically different from that in Ltk- cells or in HSV biochemically transformed cells. One peak of TK activity with an Rm value of 0.8 0.9, corresponding to mitochondrial TK, was observed on polyacrylamide gel electrophoresis of Ltk- cell extracts. VZV infected Ltk- cells had two peaks of TK activity with Rm values of 0.45-0.5 (peak I) and 0.8-0.9 (peak II). Peak I and II were concluded to be virus-specific TK and mitochondrial TK, respectively. VZV biochemically transformed cells had a peak of activity with an Rm value of 0.4 0.5, corresponding to peak I in VZV-infected Ltk- cells; that is VZV-specific TK activity. The present study indicates that VZV has a gene coding for its own TK. PMID- 6299268 TI - [The role of cAMP in the energy metabolism of human erythrocytes]. AB - In human red blood cell there is complete adenyl cyclase--cAMP system. However, the role of this system and its connection with energy metabolism in red cell is not known. Hence, we studied the effects of cAMP and DB-cAMP on energy metabolism in human red cell. Blood was taken out from haematologically healthy volunteers. Red cells were isolated by differential centrifugation and washed three times in Tris-Ringer buffer (pH 7.4). The red cells suspensions (in the above buffer) were simultaneously incubated without additions (control) and with cAMP or DB-cAMP (5 mmol/L). Incubation temperature was 37 degrees C. The samples of suspensions for the extraction of metabolites and cofactors were taken at 0 and 180 minutes of incubation. The lactate, glucose, G6P, F6P, TP, ATP, ADP and AMP were determined in neutralized perchloric acid extracts by specific enzymatic methods. Results of our experiments show that cAMP and DB-cAMP significantly increase red cell glycolysis. Under the same conditions these nucleotides induce positive "cros over" point at the phosphofructokinase (PFK) step of the glycolysis. The levels of adenine nucleotides (ATP, ADP, AMP) were unchanged through a whole period of incubation. We concluded that cAMP and DB-cAMP stimulates glycolytic process in red cells probably by allosteric activation of PFK. PMID- 6299269 TI - Eco RI restriction-site polymorphism of the albumin gene in different inbred strains of rat. AB - Two types of variant EcoRI restriction enzyme patterns of albumin-gene DNA fragments have been detected in different rat strains by agarose gel electrophoresis and Southern blot hybridization using 32P-labeled cloned rat albumin cDNA probes. The type I albumin gene variant is characteristic of the Sprague-Dawley strain, and type II is found in Buffalo rats. The occurrence of these variants is interpreted as the result of simple allelic polymorphism because they are inherited in a normal Mendelian fashion when crossing Sprague Dawley and Buffalo rats. The distribution of the two genetic variants in various inbred strains of rat suggests that type I represents the original or ancestral form of the albumin gene and that type II appeared spontaneously during laboratory breeding. PMID- 6299270 TI - Interaction of beta-iodopenicillanate with the beta-lactamases of Streptomyces albus G and Actinomadura R39. AB - The beta-lactamases of Streptomyces albus G and Actinomadura R39 are inactivated by beta-iodopenicillanate. However, in contrast with the beta-lactamase I from Bacillus cereus, they also efficiently catalyse the hydrolysis of the inactivator; with the S. albus G enzyme, kcat. is larger than 25s-1 and the number of turnovers before inactivation is 515. With the A. R39 enzyme, kcat. is larger than 50s-1 and the number of turnovers before inactivation is 80. After hydrolysis of the beta-lactam amide bond, the product rearranges into 2.3-dihydro 2,2-dimethyl-1,4-thiazine-3,6-dicarboxylate, which exhibits an absorption maximum at 305 nm. PMID- 6299271 TI - Characterization of the c-type cytochromes of Nitrosomonas europaea with the aid of fluorescent gels. AB - When a total soluble extract of Nitrosomonas europaea was denatured with dodecyl sulphate, subjected to dodecyl sulphate/polyacrylamide-gel electrophoresis and illuminated with near-u.v. light, eight bands of protein fluorescence were observed. All but one of these bands were red in colour, a property characteristic of c-type cytochromes. Standard techniques were used to purify soluble c-type cytochromes from this organism, and it was then possible to assign all but two very minor bands to specific c-type cytochromes, namely hydroxylamine oxidase, cytochrome c-554, cytochrome c-552 and a cytochrome c-550 not previously described. The eight band had fluorescence peaking in the green region of the spectrum, probably caused by covalently bound flavin, and co-purified with hydroxylamine oxidase. The following physical properties were determined for these components: isoelectric point, molecular weights according to gel filtration and mobility on dodecyl sulphate/polyacrylamide gels, and alpha-band spectra at room temperature and 77K. Redox potentials were measured as follows: cytochrome c-554, E(m,7) = +20mV; cytochrome c-552, E(m,7) = +230mV; cytochrome c 550, E(m,7) = +140mV. When washed membranes were applied to dodecyl sulphate/polyacrylamide gels in the same way, a number of fluorescent bands were observed that could be matched by soluble proteins. In addition, there was one band that could not be detected in supernatants, migrating with an apparent molecular weight of 24000. This species is probably coincident with a c-type cytochrome having E(m,7) = +170mV found in redox titration of these membranes. In future studies, gel fluorescence should form a useful complement to spectroscopy for analysis of cytochrome composition in active cell-free preparations or semi purified material. PMID- 6299272 TI - A model for the regulation of the calmodulin-dependent enzymes erythrocyte Ca2+ transport ATPase and brain phosphodiesterase by activators and inhibitors. AB - Acidic phospholipids, unsaturated fatty acids and limited proteolysis mimic the activating effect of calmodulin on erythrocyte Ca2+-transport ATPase and on brain cyclic nucleotide phosphodiesterase, as has been reported previously in several studies. Three different antagonists of calmodulin-induced activation of these enzymes were tested for their inhibitory potency on the stimulation produced by the other activators. Trifluoperazine and penfluridol were found to antagonize all the above mentioned types of activation of Ca2+-transport ATPase in the same concentration range. Both inhibitors also can reverse the activation of phosphodiesterase by oleic acid, phosphatidylserine and calmodulin at similar concentrations. However, in contrast with erythrocyte Ca2+-transport ATPase, activation of phosphodiesterase by limited tryptic digestion cannot be antagonized by penfluridol and trifluoperazine. Calmidazolium, formerly referred to as compound R 24571, was found to be a relatively specific inhibitor of calmodulin-induced activation of phosphodiesterase and Ca2+-transport ATPase, since antagonism of the other activators required much higher concentrations of the drug. The results suggest that the investigated drugs exert their inhibitory effect on calmodulin-regulated enzymes not solely via their binding to calmodulin but may also interfere directly with the calmodulin effector enzyme. In addition, a general mechanism of activation and inhibition of calmodulin-dependent enzymes is derived from our results. PMID- 6299273 TI - Superoxide dismutase-inhibitible reduction of cytochrome c by the alloxan radical. Implications for alloxan cytotoxicity. AB - Cytochrome c was reduced when superoxide was generated from xanthine oxidase in the presence of alloxan, and by the reaction of alloxan and with reduced glutathione. In each case, most of the reduction was inhibited by superoxide dismutase, but considerably more enzyme was required than with superoxide alone. This indicates that the superoxide dismutase-inhibitible cytochrome c reduction was mainly due to a direct reaction with the alloxan radical, and implies that other reactions that are inhibited by superoxide dismutase could be due to either alloxan radicals or superoxide. PMID- 6299274 TI - Comparative studies on oestrogen-induced rat uterus peroxidase and rat eosinophil peroxidase. AB - Rat eosinophil peroxidase and rat uterine peroxidase II showed similar electrophoretic mobilities, molecular weights, specific activities and spectral properties and could be purified by essentially identical techniques. Antibodies raised against the uterine enzyme strongly inhibited the eosinophil enzyme. It is suggested that rat eosinophil peroxidase and rat uterine peroxidase II may well be one and the same enzyme. PMID- 6299275 TI - Heterogeneity of the Type 3 copper in Japanese-lacquer-tree (Rhus vernicifera) laccase. AB - The two steps of the titration of the Japanese-lacquer-tree (Rhus vernicifera) laccase with N3- [Morpurgo, Rotilio, Finazzi-Agro & Mondovi (1974) Biochim. Biophys. Acta 336, 324-328; LuBien, Winkler, Thamann, Scott, Co, Hodgson & Solomon (1981) J. Am. Chem. Soc. 103, 7014-7016] were shown to be two distinct reactions, each involving one different portion of the native enzyme molecules. The difference consists in the oxidation state of the Type 3 Cu, which is reduced in the portion with higher affinity for N3- and oxidized in the portion with lower affinity for N3-. The difference is eliminated by treatment with oxidizing (H2O2) or reducing agents, and a single N3- adduct is then formed. The e.p.r. spectra of the H2O2-treated enzyme and of its F- derivatives support this interpretation of the results. The similarity of the spectroscopic properties of the high-affinity N3- adduct to those of the N3- adducts of half-met-haemocyanins and half-met-tyrosinase is discussed. PMID- 6299276 TI - Multimers of anionic amphiphiles mimic calmodulin stimulation of cyclic nucleotide phosphodiesterase. AB - Oleic acid, phosphatidylserine and pyrenedecanoic acid were found to activate calmodulin-deficient cyclic nucleotide phosphodiesterase at concentrations above their critical micellar concentration. In contrast with calmodulin these activators do not require the presence of Ca2+ for their action. It is shown that the size of phosphatidylserine vesicles is of crucial importance with respect to the activating potency of phosphatidylserine. Fluorescence measurements with the probe pyrenedecanoic acid revealed that micelles rather than monomers are the active species for stimulation of phosphodiesterase. There are indications that this result also may be applied to the other activators. PMID- 6299277 TI - A new unextracted-sample radioimmunoassay method for hepatic endogenous nuclear L tri-iodothyronine content. Validity of its use in determining nuclear receptor binding characteristics. AB - Endogenous l-tri-iodothyronine content in an hepatic nuclear extract was measured by a new unextracted-sample radioimmunoassay method using 8-anilinonaphthalene-1 sulphonic acid to inhibit the l-[(125)I]tri-iodothyronine binding to the nuclear l-tri-iodothyronine receptor within the extract. For this method, the lower sensitivity limit was 3.125 pg/tube, the recovery of added l-tri-iodothyronine was 90-120%, and the between-assay coefficient of variation was 10%. The amount of endogenous l-tri-iodothyronine was 10-40 pg/0.2 ml of hepatic nuclear extract from euthyroid rats, compared with less than 3.125 pg/0.2 ml from thyroidectomized rats. The results obtained by this new method were compared with a Sephadex G-25 column extracted-sample radioimmunoassay method and showed a good agreement. The values for the endogenous l-tri-iodothyronine content were utilized to correct for the l-tri-iodothyronine concentration within the binding assay mixture in order to accurately determine by Scatchard analysis the binding characteristics of the nuclear l-tri-iodothyronine receptor. The validity of the correction for endogeneous l-tri-iodothyronine was demonstrated by using a nuclear extract from a thyroidectomized rat which was preincubated with a small known amount of l-tri-iodothyronine before determining the nuclear l-tri iodothyronine receptor binding characteristics. When the Scatchard plots were corrected for the preincubated dose, the results obtained were similar to true values, but they were falsely lower when not corrected. It is concluded that the necessity and validity of using endogenous l-tri-iodothyronine corrections in the Scatchard analytical computations of the nuclear l-tri-iodothyronine receptor binding characteristics has been demonstrated, being particularly more important for affinity constant than maximum binding capacity. PMID- 6299278 TI - Nuclear interactions of retinoic acid-binding protein in chemically induced mammary adenocarcinoma. AB - Cellular retinoic acid-binding protein (CRABP) was detected in the nuclear fraction of N-methyl-N-nitrosourea-induced mammary cancers after the incubation of cytosol containing [3H]retinoic acid (RA)-bound CRABP with isolated nuclei. CRABP extracted from the nuclei in buffer containing 0.4 M-KCl sedimented as a 2 S component when subjected to sucrose-density-gradient analysis. [3H]RA-CRABP was found to be a prerequisite for the detection of nuclear binding, since the incubation of isolated nuclei or 0.4 M-KCl extract of the nuclei with [3H]RA did not result in any significant binding. Incubation of [3H]RA-CRABP at 25 or 30 degrees C before incubation with the nuclei neither altered the sedimentation coefficient nor enhanced the nuclear binding compared with 0 degrees C incubation. The tumour nuclei contained a saturable number of binding sites with a dissociation constant of 1.6 x 10(-9) M. These results indicate that the action of retinoic acid in the target organ may be mediated by its interaction with the nuclei. PMID- 6299279 TI - Abundant amounts of diadenosine 5',5"'-P1,P4-tetraphosphate are present and releasable, but metabolically inactive, in human platelets. AB - Diadenosine 5',5"'-P1,P4-tetraphosphate (Ap4A) may be formed in the back reaction of the amino acid-activation reaction [Zamecnik, Stephenson, Janeway & Randerath (1966) Biochem. Biophys. Res. Commun. 24, 91-98]. On the basis of a number of observations of the properties of Ap4A it has been suggested that it may have a signal function for the initiation of DNA replication in eukaryotic cells] Grummt (1978) Proc. Natl. Acad. Sci. U.S.A. 75, 371-375]. In the present paper human platelets have been shown to contain relatively large amounts of Ap4A. The compound is apparently metabolic inactive in platelets, but it is almost quantitatively released when platelets are activated to aggregate by treatment with thrombin. The results are discussed in connection with the known growth stimulating activity of platelets. PMID- 6299281 TI - Activation of glycerophosphocholine phosphodiesterase in rat forebrain by Ca2+. AB - The highest activity of glycerophosphocholine phosphodiesterase (EC 3.1.4.2) in subcellular fractions of rat forebrain was found in the microsomal fraction though significant amounts were found in fractions containing plasma membranes. With the use of Ca(2+)/EGTA and Ca(2+)/EDTA buffers it was shown that very low concentrations of free Ca(2+) (EC(50)approx. 10(-9)m) could activate the enzyme. PMID- 6299280 TI - The liver angiotensin receptor involved in the activation of glycogen phosphorylase. AB - Specific angiotensin binding to rat hepatocytes and purified liver plasma membranes was measured by using biologically active [(3)H]angiotensin (sp. radioactivity 14Ci/mmol). The kinetic parameters for angiotensin binding to hepatocytes are: K(+1) (association rate constant). 100mum(-1).min(-1); K(-1) (dissociation rate constant), 2min(-1); K(d) (dissociation constant). 30nm; maximal binding capacity, 0.42pmol/10(6) cells or 260000 sites/cell. Angiotensin binding to membranes is profoundly affected by GTP (0.1mm) and NaCl (100mm); these regulatory compounds greatly enhance both the rate of association and of dissociation and also the extent of dissociation. K(d) amounts to 10nm in the presence of GTP+NaCl and to 1.5nm in their absence; maximal binding capacity is 0.70pmol/mg of protein, both with or without GTP+NaCl. The relative affinities of 11 angiotensin structural analogues were deduced from competition experiments for [(3)H]angiotensin binding to hepatocytes and to membranes (in the latter case, GTP + NaCl were not included, in order to study the higher affinity state of the receptor). These are highly correlated with their biological activity (activation of glycogen phosphorylase in hepatocytes). Binding to membranes occurs in the same concentration range as the biological effect. On the other hand, the existence of numerous spare receptors is suggested by the observation that binding of the agonists to hepatocytes requires 25-fold higher concentrations than those needed for their biological activity. These data clearly suggest that the detected binding sites correspond to the physiological receptors involved in the glycogenolytic action of angiotensin on rat liver. PMID- 6299282 TI - Reversal of nonresponsiveness to bacterial endotoxins in C3H/HeJ mice by glucan and streptozotocin. PMID- 6299283 TI - Effects of vanadate on heart protein kinase. PMID- 6299284 TI - Desensitization of beta 2-adrenergic receptors and adrenocorticotropin release. AB - Pre-exposure of mouse anterior pituitary tumor cells (A+T-20/D16-16) to (-) isoproterenol reduces the ability of this beta-adrenergic agonist to restimulate cyclic AMP synthesis or adrenocorticotropin hormone (ACTH) release from these cells. This beta-adrenergic receptor desensitization is time and dose-dependent, recoverable and specific for beta-receptors. Longer pretreatment times are required to decrease beta-receptor density than to induce receptor desensitization. This initial beta-receptor refractoriness involves an uncoupling of the receptor from adenylate cyclase since (-) isoproterenol treatment does not alter forskolin-activated cyclic AMP formation or ACTH release. In addition to diminishing beta-receptor responsiveness, (-) isoproterenol treatment induces a prolonged elevation of basal ACTH release. This finding indicates that the intracellular events leading to ACTH secretion may also be altered during the desensitization process. PMID- 6299285 TI - Evidence for the existence of gonadotropin receptors in the nuclei isolated from rat ovary. AB - Specific binding of radiolabeled human chorionic gonadotropin (hCG) to nuclei isolated from pseudopregnant rat ovaries was studied. Incubation of cultured luteal cells or isolated nuclei with fluorescein isothiocyanate conjugated hCG showed concentration of fluorescence in the nuclear region. Isolated nuclei exhibited saturable high affinity binding of radiolabeled hCG with an apparent Kd of 3.42 X 10(-10) M. The binding was inhibited by increasing concentrations of unlabeled hCG. Under dissociating conditions, the bound hCG was dissociated from the nuclei. However, unlike the plasma membranes, the hCG bound to nuclei was not degraded before dissociation. Radiolabeled hCG bound to the nuclei could also be dissociated by brief exposure to MgCl2 or acidic incubation medium. The bound hCG was not extractable with 4M KCl or 2% Triton X-100. The available evidence suggest that nuclear receptors are distinct from plasma membrane receptors for hCG. PMID- 6299286 TI - Dipeptide mimics. Conformationally restricted inhibitors of angiotensin converting enzyme. PMID- 6299287 TI - pH dependence of dissociation of the ovine prolactin rabbit mammary receptor complex. AB - Ovine prolactin specifically bound to rabbit mammary membrane prolactin receptor was rapidly dissociated in a pH dependent manner with 0.1M ammonium acetate. Up to 75% of the bound hormone was dissociated at pH 4.0 or lower in less than 5 minutes. The pK of the dissociation was 4.7, implicating one or more critical carboxyl groups. Exposure of the membrane bound receptor to the dissociating buffer for up to one hour did not reduce its ability to bind hormone. The dissociated hormone was characterized as intact ovine prolactin by Bio-Gel P-150 gel chromatography and by its ability to bind to fresh rabbit prolactin receptor with the same binding affinity as native hormone. PMID- 6299288 TI - Reduction of cytochrome c by hydrogen peroxide and its inhibition by superoxide dismutase. AB - Ferricytochrome c is slowly converted by hydrogen peroxide to an equilibrium mixture of ferricytochrome c and ferrocytochrome c, and in the process, the hydrogen peroxide is decomposed. The reductant appears to be superoxide anion, produced from the reaction of hydrogen peroxide with oxygen. Because the reduction of ferricytochrome c by hydrogen peroxide is inhibited by superoxide dismutase, we propose that the enzyme acts by converting superoxide anion to a dimerized product that is less active as a reductant. PMID- 6299289 TI - Modification of the sensitivity of CHO cells to mitomycin C by dibutyryl cyclic AMP. AB - The survival of CHO cells exposed to mitomycin C was decreased three times that of the cells treated with 1 mM dibutyryl cyclic AMP before mitomycin C treatment, as compared to the absence of treatment with this cyclic nucleotide. The sensitization effect began at 3-4 hours after the start of pre-treatment, reached a maximum at around 10 hours and continued to be effective. Post-treatment with the cyclic nucleotide for more than 12 hours increased the survival of CHO cells exposed to mitomycin C. PMID- 6299290 TI - Action of peptidases in brain synaptic membranes on the NH2-terminus of adrenocorticotropin using ACTH-(1-16)-NH2 as a model substrate. AB - The action of brain peptidases on NH2-terminal sequences of adrenocorticotropin was studied by incubation of ACTH-(1-16)-NH2 under different pH conditions. Profiles of metabolites and time course of product formation were obtained by HPLC analysis of the digests. Fragments of ACTH-(1-16)-NH2 were isolated and characterized by their amino acid composition and NH2-terminal groups. Both at pH 7.4 and pH 8.5 the following fragments were found: ACTH-(3-16)-NH2, ACTH-(4-16) NH2, ACTH-(5-16)-NH2, and ACTH-(7-16)-NH2. At pH 7.4 the major products were ACTH (4-16)-NH2 and ACTH-(7-16)-NH2, while the peptide ACTH-(3-16)-NH2 was the main metabolite at pH 8.5. The nature of identified peptides and the time course of their formation demonstrates that aminopeptidase activities predominate in the conversion of the NH2-terminus of adreno-corticotropin and related peptides by brain synaptic membranes. PMID- 6299291 TI - Dark-light regulation of pyruvate, Pi dikinase in C4 plants: evidence that the same protein catalyses activation and inactivation. PMID- 6299292 TI - Homology between actin coding and its adjacent sequences in widely divergent species. AB - Eco RI restriction endonuclease DNA fragments from several representatives of the kingdoms Protista and Animalia were electrophoretically separated and transferred to the nitrocellulose filters. These DNA's were hybridized with [32p]-labelled actin coding sequence from Drosophila melanogaster (Dm). The results indicate that the nucleic acid sequences of the genes coding for actin(s) has been highly conserved throughout evolution. Similar experiments were performed using the sequence derived from the 5' end of Drosophila actin gene as a probe. Cross hybridization was observed between Drosophila and Acanthamoeba castellanii. This may indicate a functionally important region at the 5' end which has been conserved. PMID- 6299293 TI - Mestranol-induced hypertension: characterization of cytochrome P-450 dependent catechol estrogen formation in brain microsomes. PMID- 6299294 TI - Role of nicotinamide adenine dinucleotide in ethanol-induced depressions in testicular steroidogenesis. AB - It is rapidly becoming accepted, without direct evidence, that a change in the NAD+/NADH ratio in the testes produced by the metabolism of ethanol is the principal mechanism involved in its now well-established effects on testicular steroidogenesis. The purposes of the present studies were 2-fold: (1) to examine whether, in fact, in vivo or in vitro ethanol exposure alters the NAD+/NADH ratio in the testes; and (2) to examine the validity of previous reports in which it was found that NAD+ prevented the effects of ethanol on testicular steroidogenesis under in vitro conditions. With regard to the first objective, we found that a large dose of ethanol (2.5 g/kg) markedly reduced gonadotropin stimulated testicular steroidogenesis in vivo in the male rat, but it did not alter the NAD+ and NADH concentrations in the testes. Similarly, extremely high ethanol concentrations (200 mM) substantially suppressed hMG-stimulated testosterone biosynthesis in in vitro Leydig cell preparations but no change in NAD+ concentration occurred; NADH levels were very low in the Leydig cell preparations (less than 2% of NAD+ levels), but did not appear to change as a function of ethanol exposure. Finally, in contrast to previously published results, we found that NAD+ (1 mM) did not prevent the in vitro effects of ethanol on cAMP-stimulated testicular steroidogenesis. Consequently, our results fail to support the hypothesis that acute in vivo or in vitro ethanol administration inhibits the biosynthesis of testosterone by altering the NAD+/NADH ratio in the testes. PMID- 6299295 TI - Metabolism of polycyclic aromatic hydrocarbons in the rat ovary. Comparison with metabolism in adrenal and liver tissues. PMID- 6299296 TI - Kinetic parameters of 3',5'-cAMP phosphodiesterase in cat sciatic nerve. PMID- 6299297 TI - Enhanced binding of the convulsive ligand DMCM to high-energy irradiated benzodiazepine receptors; evidence of complex receptor structure. PMID- 6299298 TI - Non-enzymatic degradation of acid-soluble calf skin collagen by superoxide ion: protective effect of flavonoids. AB - Calf skin acid-soluble collagen in microfibrillar form was incubated with free oxygen radicals produced by the system xanthine oxidase + hypoxanthine. This incubation liberated peptides of a size smaller than that of alpha-chains, as demonstrated by SDS-PAGE and by evaluation of the 4-hydroxyproline contained in small peptides. The amount of liberated peptides was found to increase with time. The process was inhibited by addition of superoxide dismutase to the medium but not by addition of catalase. Two flavonoids extracted from bilberries and a third one from grapes were demonstrated to protect collagen against this non-enzymatic proteolytic activity. This work confirms that collagen may be degraded during the process of inflammation and that some flavonoids are endowed with protective properties. PMID- 6299300 TI - Cardiovascular consequences of endogenous opiate antagonism. PMID- 6299299 TI - Guanine nucleotides and alpha 1 adrenergic receptors in the heart. PMID- 6299301 TI - EPR study on the interaction of spin-labelled hydrazine mustard derivatives with DNA. AB - The interaction of three spin-labelled compounds, derivatives of bis-(2 chloroethyl)-hydrazine (HMSL), N-methyl,N-chloroethyl-hydrazine (MCEHSL) and bis (2-bromoethyl)-hydrazine (BEHSL) with DNA was studied by the method of electron paramagnetic resonance (EPR). It was found that HMSL (containing two chloroethyl groups) and MCEHSL (containing one chloroethyl group) gave spin-labelled dsDNA with identical strongly immobilized EPR spectra. The conclusion was drawn that only one of the alkylating groups of HMSL reacted with DNA. In contrast, the EPR spectrum of DNA spin-labelled with BEHSL was non-immobilized due to the strong destabilizing effect of this compound on the double helix. The extent of alkylation of DNA with the three hydrazine mustard derivatives was one and the same. It was found, however, that chloroethyl-containing derivatives (HMSL and MCEHSL) had an expressed base specificity and alkylated preferably the guanilic residues, and their bromo-analogue (BEHSL) did not show any base specificity and alkylated the bases of DNA at random. PMID- 6299302 TI - Brain receptor binding and lipophilic character of benzodiazepines. AB - A series of measurements of the potency of 26 benzodiazepines (BDZs) to inhibit the binding of [3H]diazepam to rat brain synaptosomal membranes has been undertaken. All compounds studied are NO2 or Cl 7-substituted, a class of BDZs known to have the highest biological activities and binding affinities (expressed as Ki). The results show that: (a) 2'-unsubstituted BDZs display a parabolic dependence of -log Ki values on lipophilic character of the molecule (expressed by means of the chromatographic Rm value); (b) 2'-halogen substituted BDZs show quite high binding affinities (ki values in the range 2-5 nM) giving rise to another class of BDZs whose dependence on the lipophilic character remains to be studied; and (c) BDZs lacking the carbonyl oxygen at position 2 (see Table 1), and having an oxygen at position 4, show low or very low binding affinities (Ki values in the range 600-7000 nM). Moreover for the compounds under examination, statistically significant correlations have been obtained between -log Ki values and psychopharmacological activity data. PMID- 6299303 TI - Effect of ACTH on cytochrome P-450 content and DMBA metabolism in immature rat adrenal. PMID- 6299305 TI - Effects of acute and chronic desmethylimipramine on levels of cyclic AMP in vivo. PMID- 6299304 TI - Cyclic AMP induced stimulation and inhibition of Ca2--uptake in rat cardiac sarcolemma vesicles. PMID- 6299306 TI - Quantitative correlation of cardiac glycoside binding to its receptor and inhibition of the sodium pump in chicken heart cells in culture. PMID- 6299307 TI - Characterization of the vasodilator action of antihypertensive drug budralazine. AB - Intravenous injection of 1-[2-(1,3-dimethyl-2-butenylidene)hydrazino]phthalazine (budralazine) to anesthetized dogs resulted in an increase in cardiac output and regional blood flow in various vascular beds, and a fall in mean blood pressure with a decreased total and regional vascular resistance. Budralazine produced a dose-related increase in femoral blood flow in anesthetized dogs when injected into the femoral artery. The vasodilator drug relaxed either KCl(K+)- or noradrenaline (NA)-induced contractions of isolated rabbit aorta in a concentration-dependent manner. In the K+-depolarized aorta, it also produced a concentration-related inhibition of contractile response to cumulative addition of Ca2+. Intra-arterial injection of budralazine dilated dose-dependently the isolated perfused vascular bed of rabbit ear constricted by either K+ or NA. Budralazine, at effective antihypertensive oral dose, was without significant effect on cyclic nucleotide levels in the aorta of spontaneously hypertensive rats (SHR). These results indicate that budralazine, like hydralazine, produces vasodilation through a direct effect on vascular smooth muscle which may result at least in part from its inhibitory effect on vascular Ca2+ fluxes. PMID- 6299308 TI - Effect of ethacrynic acid on mitochondrial electron transport system and oxidative phosphorylation. PMID- 6299309 TI - Glucuronic acid conjugate of delta 1-tetrahydrocannabinol identified in the urine of man. AB - A delta 1-tetrahydrocannabinol (delta 1-THC) metabolite has been identified as an O-glucuronide of delta 1-THC in the human urine after oral administration of delta 1-THC. The metabolite was present in small amounts and its identity was confirmed by comparison of its mass spectrum to that of a reference compound. PMID- 6299310 TI - [Mezlocillin: a penicillin from the acylureido series. Synthesis and chemical properties]. AB - Mezlocillin, 6-[D-2-(2-Oxo-3-mesyl-imidazolidine-1-carboxamido)- 2 phenylacetamido]-penicillanic acid, is a new semisynthetic acylureido-penicillin with a broad spectrum of antibacterial activity. The synthesis of mezlocillin from ampicillin and from 6-aminopenicillanic acid and the physicochemical properties are described. Mezlocillin is cleaved by penicillinase to the penicilloate which is degraded further by acids to the penilloate. PMID- 6299311 TI - Sodium, and potassium dependent adenosinetriphosphatase from Cavia cobaya kidney: interaction with very low cardiotonic steroid concentrations. AB - We noticed that very low cardiotonic steroid concentrations activate the Na, K ATPase in a variety of different preparations. In the present research the effect of three cardioactive steroids on the enzymatic activity was tested. The glycosides activated the Na,K-ATPase, while the aglycone strophantidine does not. Ouabain binding studies on various preparations showed the presence of two binding site classes with different affinities. Purification procedures shift the apparent Kd values, while K+ increase them. Accordingly, the activatory and inhibitory effects may be explained by the cardiotonic steroid binding on different sites of the Na,K-ATPase molecule. PMID- 6299312 TI - Na,K-ATPase from Xenopus laevis kidney and epidermis: ouabain interaction studies. AB - A satisfactory purification from Xenopus laevis epidermis is presented. Ki and Kd values for the ouabain-enzyme interaction have been evaluated. Both parameters appear very low, as compared with those demonstrated in other anurans. Very low digitalis-like compound level was demonstrated in Xenopus; on the contrary in Bufo, in which these substances play a defensive role, it is very high. Our results strengthen the hypothesis of a physiological action of such compounds in regulating enzyme-driven Na+/K+ exchanges. PMID- 6299313 TI - [Changes in Na+, K+-adenosinetriphosphatase and 5'-nucleotidase activities in cell membrane isolated from rat liver after acute white phosphorus poisoning]. AB - Na+, K+-ATPase and 5'-Nucleotidase activities in rat liver plasmamembranes after "in vivo" intoxication with a single dose of white phosphorus (10 mg/kg b.w. "per os") are investigated. Na+, K+-ATPase activity is significantly increased 1 hour and inhibited 12 hours after intoxication. 5'-Nucleotidase is strongly increased at 1, 2 and 4 hours after poisoning and is significantly decreased at 12 hours. The enhancement of both the enzymatic activities is evident prior to triglyceride accumulation in rat liver. Our results suggest that lipid fluidity of cell membrane is early and mildly affected during white phosphorus poisoning. PMID- 6299316 TI - Mechanisms of general anaesthesia. PMID- 6299315 TI - Value of electrocardiogram in diagnosing right ventricular involvement in patients with an acute inferior wall myocardial infarction. AB - To study the value of the electrocardiogram in diagnosing right ventricular involvement in acute inferior wall myocardial infarction, the electrocardiographic findings were analysed in 67 patients who had had scintigraphy to pin-point the infarct. All 67 patients were consecutively admitted because of an acute inferior wall infarction. A 12 lead electrocardiogram with four additional right precordial leads (V3R, V4R, V5R, and V6R) was routinely recorded on admission and every eight hours thereafter for three consecutive days. Thirty-six to 72 hours after the onset of chest pain a 99mtechnetium pyrophosphate scintigraphy and a dynamic flow study were performed to detect right ventricular involvement, which was found in 29 of the 67 patients (43%). ST segment elevation greater than or equal to 1 mm in leads V3R, V4R, V5R, and V6R is a reliable sign of right ventricular involvement. ST segment elevation greater than or equal to 1 mm in lead V4R was found to have the greatest sensitivity (93%) and predictive accuracy (93%). The diagnostic value of a QS pattern in lead V3R and V4R or ST elevation greater than or equal to 1 mm in lead V1 was much lower. ST segment elevation in the right precordial leads was short lived, having disappeared within 10 hours after the onset of chest pain in half of our patients with right ventricular involvement. When electrocardiograms are recorded in patients with an acute inferior wall infarction within 10 hours after the onset of chest pain, additional right ventricular infarction can easily be diagnosed by recording lead V4R. PMID- 6299314 TI - Converting enzyme inhibitors in hypertension and heart failure. PMID- 6299317 TI - Human lymphoblastoid interferon in the treatment of small cell lung cancer. AB - Ten patients with small cell lung cancer were treated with high dose human lymphoblastoid interferon (50-100 megaunits m-2) for 5 days, followed by low dose interferon (3 megaunits m-2) for 3 weeks. At the end of treatment, and one month later, there was no evidence of either complete or partial response. The treatment produced fever, anorexia and weight loss, with transient leucopenia and thrombocytopenia; there was evidence of a non-cholestatic elevation of serum alanine aminotransferase, with clinical deterioration in the condition of three patients presenting with hyponatraemia. A transient hypocalcaemia during high dose therapy was also noted. It seems that lymphoblastoid interferon as a single agent is unlikely to have a role in the treatment of small cell lung cancer, and that its administration as employed in this study is associated with considerable toxicity. PMID- 6299318 TI - Second primary malignancies following salivary gland cancers. AB - Four hundred and fifteen males and 367 females who had invasive malignant tumours of the salivary glands as their first cancer diagnosed in Connecticut between 1935 and 1978 were identified and followed 2342 and 2868 person-years respectively. Overall a slight excess of second primary cancers (relative risk 1.35) was observed. Significant excesses were noted for respiratory cancers in males (relative risk 2.8) and for ovarian cancer (relative risk 5.3) but not breast cancer (relative risk 1.3) in women. Possible reasons for excesses at these sites are discussed, but it seems most likely they are related to small number variation. PMID- 6299319 TI - Radioprotection by WR-2721 in vitro at low oxygen tensions: implications for its mechanisms of action. AB - Radioprotection of spheroids of Chinese hamster V79 cells by WR-2721 was found to be a function of spheroid size, with the greatest dose-modifying effect by the protector observed for spheroids almost large enough to contain radioresistant "anoxic" cells. The nature of the response suggested that most of the protective effect was due to the presence of an increased hypoxic fraction in the drug treated spheroids. Similarly, when single-cell suspensions were irradiated at various oxygen tensions, one component of radioprotection by WR-2721 was found to be highly dependent upon the available oxygen. Two mechanisms of radioprotection of V79 cells by WR-2721 were thus demonstrated: a modest, oxygen-independent effect, presumably due to hydrogen donation, and an oxygen-depleting effect, which is of maximal significance for cells or tissues which would otherwise be partially sensitized by low levels of oxygen. PMID- 6299320 TI - Carcinoembryonic antigen (CEA) in explants of human breast cancer: comparison of immunohistochemical detection and release during short-term culture. PMID- 6299321 TI - Familial sea-blue histiocytosis with cutaneous involvement. A case report with ultrastructural findings. PMID- 6299322 TI - Hepatic toxicity of ketoconazole. PMID- 6299324 TI - Rhesus immunization in male volunteers: changes in lymphocyte functions following secondary immunizations in anti-D responders and non-responders. AB - After secondary immunizations of rhesus(D)-negative male volunteers with Rh(D) positive red cells, changes were found in the in vitro transformation and antibody-dependent cell-mediated cytotoxicity (ADCC) capacities of the volunteers' lymphocytes. Responders, who produced anti-D, showed marked depressions of ADCC which were not found in non-responders. Responders and non responders in general showed similar changes in lymphocyte transformation. The relationships between altered lymphocyte functions following immunization, immunoregulatory activity and responsiveness to the Rh(D) antigen are discussed. PMID- 6299323 TI - Heparin-binding platelet proteins demonstrated by crossed affinity immunoelectrophoresis. AB - Platelet proteins that interact with heparin were studied using crossed affinity immunoelectrophoresis. Platelet proteins solubilized in Triton X-100 were applied to crossed immunoelectrophoresis against anti-platelet antibodies, and an intermediate gel containing heparin covalently linked to Sepharose 4B was inserted. Six immunoprecipitates were absent or showed an altered position compared to control immunoplates, indicating that the corresponding antigens were bound to the immobilized heparin. These precipitates represented platelet factor 4, thrombospondin, glycoprotein Ib, and three antigens termed G4, 17 and 25. The subcellular location of the heparin-binding proteins was either in the surface membrane (glycoprotein Ib and the antigens 17 and 25), or in the alpha-granules (platelet factor 4, thrombospondin and G4). both forms of platelet factor 4 appearing after crossed immunoelectrophoresis, i.e. a line-form and a peak-form, bound strongly to the heparin. Glycoprotein Ib showed a weak binding whereas its proteolytic split product glycocalicin did not significantly bind to the heparin in the present system. It is concluded that the platelets contain at least six heparin-binding proteins which are present on the cellular surface or are able to be exposed to the extracellular medium after the release-reaction has occurred. PMID- 6299325 TI - Alpha globin gene analysis in a Sardinian family with interacting alpha and beta thalassaemia genes. AB - This paper reports the results of alpha globin gene analysis in a Sardinian family with interacting alpha and beta thalassaemia genes. The propositus, who was identified in a newborn survey as he had 26.0% Hb Bart's and 74.0% Hb F, successively developed the clinical and haematological picture of a transfusion dependent thalassaemia major. According to the haemoglobin pattern, restriction endonuclease analysis of the DNA from this patient showed the deletion of three of the four alpha-globin structural genes. Thus beta 0-thalassaemia homozygotes with the delection of three alpha-structural genes seem to have a severe clinical phenotype similar to that of patients with a full complement of four alpha-globin structural genes. PMID- 6299326 TI - Asbestos-related mesothelioma: factors discriminating between pleural and peritoneal sites. AB - Up to the end of 1980, 144 confirmed cases of mesothelioma were identified among employees of an organisation using asbestos in manufacturing and insulation. The primary site was peritoneal in 74 cases, pleural in 66, and undetermined in four. All employees had been exposed to amphibole asbestos, and evidence from different factories confirmed the predominant role of crocidolite in the production of mesothelioma. The ratio of pleural to peritoneal sites showed a continuous change when related to the year of first exposure, varying from 5:1 pleural to peritoneal before 1921 to 1:3 after 1950. The strong temporal relationship appeared to reflect progressive dust suppression, including the non-fibrous dusts present in insulation materials and perhaps also the degree to which the fibres had been opened. Other predisposing factors were related to the degree of individual exposure, the peritoneal site being associated preferentially with longer and heavier exposures. PMID- 6299327 TI - Chronic peripheral neuropathy in workers with previous exposure to carbon disulphide. AB - Two groups of viscose rayon production workers were examined 10 years after discontinuation of exposure to CS2. Group A comprised 20 subjects exposed to high CS2 concentrations and group B 13 subjects with lower exposure. Clinical findings and the measurements of nerve conduction velocity of the slow fibres in the peripheral nerves were evaluated. Twelve subjects had both clinical and electromyographic evidence of neuropathy and 10 showed a diminution in motor conduction velocity. A relationship between the degrees of exposure to CS2 and prevalence of polyneuropathy was found. On re-examining 12 subjects with neuropathy who had been examined four years before no significant electromyographic improvement was observed. These findings are consistent with a permanent axonal neuropathy caused by carbon disulphide. PMID- 6299328 TI - Neonatal cytomegalic inclusion disease in a set of twins one member of whom was a hydropic stillbirth the other completely uninfected. Case report. PMID- 6299329 TI - Reduced plasma half-life of radio-labelled 25-hydroxyvitamin D3 in subjects receiving a high-fibre diet. AB - 1. The plasma disappearance of 3H-labelled 25-hydroxyvitamin D3 (25(OH)D3) was studied in healthy volunteers on normal and high-fibre diets, using 3H-labelled tracer doses given intravenously. 2. The mean (+/- SEM) plasma half-life in the high-fibre-diet group was 19.2 +/- 1.7 d, which was significantly shorter than in the group on normal diets (27.5 +/- 2.1 d, P less than 0.01). 3. This finding suggests that a high-fibre diet leads to enhanced elimination of 25(OH)D3 by an action within the intestinal lumen. This may involve interference with an enterohepatic circulation of the metabolite, perhaps by binding of 25(OH)D3 to dietary fibre. 4. The reduced plasma half-life of 3H-labelled 25(OH)D3 associated with a high-fibre diet may explain the development of vitamin D deficiency in Asian immigrants with normal exposure to u.v. light. PMID- 6299330 TI - Substrate specificity and kinetic characteristics of angiotensin converting enzyme. AB - Furanacryloyl-Phe-Gly-Gly has been shown to be a convenient substrate for angiotensin converting enzyme (dipeptidyl carboxypeptidase, EC 3.4.15.1). A detailed kinetic analysis of the hydrolysis of this substrate indicates normal Michaelis-Menten behavior with kcat = 19000 min-1 and KM = 3.0 x 10(-4) M determined at pH 7.5, 25 degrees C. The enzyme is inhibited by phosphate and activated by chloride; maximal activity is observed with 300 mM NaCl. In the absence of added zinc, activity is lost rapidly below pH 7.5 due to spontaneous dissociation of the metal, but in the presence of zinc, the enzyme remains fully active to about pH 6. The pH-rate profile indicates two groups on the enzyme with apparent pK values of 5.6 and 8.4. The substrate specificity of the enzyme has been examined in terms of the fundamental specificity quantity kcat/KM as well as the separate constants by using a series of furanacryloyl-tripeptides. The activity toward furanacryloyl-Phe-Gly-Gly has been compared with that toward the physiological substrates angiotensin I and bradykinin. PMID- 6299332 TI - Purification and characterization of two protein kinases associated with Rous sarcoma virus. AB - The two major phosvitin-utilizing kinases have been purified from virions of the Prague C strain of Rous sarcoma virus by the use of ion-exchange and affinity chromatography. The two kinases isolated may be differentiated by their molecular weights as well as by their ability to utilize GTP as a phosphate donor. Protein kinase G, which will use either GTP or ATP as a phosphate donor, has a molecular weight of 120 000 as determined under nondenaturing conditions by glycerol gradient centrifugation and 28 000 when assayed under denaturation in sodium dodecyl sulfate (Na-DodSO4)-polyacrylamide gels. Protein kinase A, which will only efficiently use ATP as the phosphate donor, has an apparent molecular weight of 43 000 estimated by glycerol gradient sedimentation and 40 000 by NaDodSO4 polyacrylamide electrophoresis. Both kinases possess the ability to autophosphorylate. Phosvitin is the major, and casein the minor, phosphate accepting substrate for both kinases in vitro; however, kinase G will also phosphorylate histones to an extent similar to that observed with casein. PMID- 6299331 TI - Activation of angiotensin converting enzyme by monovalent anions. AB - The angiotensin converting enzyme catalyzed hydrolysis of furanacryloyl-Phe-Gly Gly is activated by monovalent anions in the order C1- greater than Br- greater than F- greater than NO3- greater than CH3COO-. In the alkaline pH region, increasing anion concentrations decrease the KM but do not change the kcat. This behavior is characteristic of an ordered bireactant mechanism in which the anion binds to the enzyme prior to the substrate. At acidic pH values, however, the anion activation is a result of both a decrease in KM and an increase in kcat, implying a bireactant mechanism in which anion and substrate bind randomly. For both the ordered and the bireactant mechanisms the anion serves as an essential activator. The effect of chloride on enzyme activity was studied over the pH range 5-10 under kcat/KM conditions and demonstrates that the apparent chloride binding constant increases from 3.3 mM at pH 6.0 to 190 mM at pH 9.0. The kcat vs. pH profile exhibits two pK values of 5.6 and 9.6, while the variation of KM with pH is characterized by a pK of 8.9 and a 2-fold increase between pH 6.5 and 7.5. The chloride activation of the hydrolysis of furanacryloyl-Phe-Gly-Gly is compared with that of the physiological substrates angiotensin I and bradykinin. PMID- 6299333 TI - Purification and properties of dihydroorotate oxidase from Crithidia fasciculata and Trypanosoma brucei. AB - Dihydroorotate oxidases have been highly purified from the parasitic protozoa Crithidia fasciculata and Trypanosoma brucei. The Crithidia enzyme was purified 4200-fold from a crude soluble protein extract in four steps. The protein is a dimer as judged from the native (Mr 60 000) and subunit (Mr 32 700) molecular weights. The purified enzyme exhibits a characteristic flavin electronic spectrum, and each mole of native dimer contains 1.0 mol of tightly bound flavin mononucleotide. Under anaerobic conditions, the flavin chromophore is reduced upon addition of L-dihydroorotate. In air-saturated buffer, the enzyme catalyzes the conversion of L-dihydroorotate to orotate with concomitant reduction of equimolar amounts of molecular oxygen to hydrogen peroxide. A variety of low molecular weight oxidants (e.g., quinones or ferricyanide) may replace oxygen as the electron acceptor during catalysis. The dihydroorotate oxidase of T. brucei was purified 1400-fold to apparent homogeneity by a highly similar isolation procedure. The estimated native (Mr 62 000) and subunit (Mr 30 500) molecular weights indicated a dimeric protein comparable in size to the enzyme from Crithidia. These results suggest that dihydroorotate oxidation is mediated by flavoprotein oxidases in these parasitic protozoa rather than by pterin-linked hydroxylases as recently proposed [Kidder, G. W., & Nolan, L.L. (1973) Biochem. Biophys. Res. Commun. 53, 929-936; Gutteridge, W. E., Dave, D., & Richards, W. H. G. (1979) Biochim. Biophys. Acta 582, 390-401]. PMID- 6299334 TI - Chromatin structure of the ovalbumin gene family in the chicken oviduct. PMID- 6299335 TI - H1 histone kinases from nuclei of Physarum polycephalum. PMID- 6299336 TI - Human skin fibroblast procollagenase: mechanisms of activation by organomercurials and trypsin. AB - Pure human skin fibroblast procollagenase has been utilized in this study as a model system in which to examine the pathways of organomercurial and trypsin activation. Three organomercurials, p-(hydroxymercuri) benzoate, mersalyl, and p aminophenylmercuric acetate, were able to fully activate human skin procollagenase with no accompanying loss of molecular weight. Lower molecular weight species were subsequently produced, particularly with a fourth organomercurial, phenylmercuric chloride. The activation process was dependent upon the concentration of the organomercurial compound and the time of incubation, but not on enzyme protein concentration. No evidence of a role for free sulfhydryls was found. Trypsin produced an initial cleavage product of procollagenase which was collagenolytically inactive yet underwent a concentration independent autocatalysis. Thus, procollagenase appeared to have an autocatalytic property which was enhanced by treatment with a variety of agents, all of which may function by perturbation of the zymogen conformation. PMID- 6299338 TI - Inhibition of gastric (H+ + K+)-ATPase by the substituted benzimidazole, picoprazole. AB - The substituted benzimidazole, picoprazole, inhibited the gastric (H+ + K+) ATPase in a concentration-and time-dependent manner. Half-maximal inhibition of the (H+ + K+)-ATPase activity was obtained at about 2 . 10(-6)M under standard conditions. In addition to the inhibition of ATPase activity, parallel inhibition of phosphoenzyme formation and the proton transport activity were achieved. Radiolabelled picoprazole was found to bind to 100 kDa peptide; this peptide was shown by phosphorylation experiments to contain the catalytic centre of the (H+ + K+)-ATPase. Studies on the (Na+ + K+)-ATPase indicated that this enzyme was unaffected by picoprazole. From the data presented and from other pharmacological studies, it is proposed that this compound inhibits acid secretion at the level of the parietal cell by its ability to inhibit the gastric proton pump, the (H+ + K+)-ATPase. PMID- 6299337 TI - The effect of pH, ubiquinone depletion and myxothiazol on the reduction kinetics of the prosthetic groups of ubiquinol:cytochrome c oxidoreductase. AB - (1) The kinetics of the reduction by duroquinol of the prosthetic groups of QH2:cytochrome c oxidoreductase and of the formation of ubisemiquinone have been studied using a combination of the freeze-quench technique, low-temperature diffuse-reflectance spectroscopy, EPR and stopped flow. (2) The formation of the antimycin-sensitive ubisemiquinone anion parallels the reduction of both high potential and low-potential cytochrome b-562. (3) The rates of reduction of both the [2Fe-2S] clusters and cytochromes (c + c1) are pH dependent. There is, however, a pH-dependent discrepancy between their rate of reduction, which can be correlated with the difference in pH dependencies of their midpoint potentials. (4) Lowering the pH or the Q content results in a slower reduction of part of the [2Fe-2S] clusters. It is suggested that one cluster is reduced by a quinol/semiquinone couple and the other by a semiquinone/quinone couple. (5) Myxothiazol inhibits the reduction of the [2Fe-2S] clusters, cytochrome c1 and high-potential cytochrome b-562. (6) The results are consistent with a Q-cycle model describing the pathway of electrons through a dimeric QH2:cytochrome c oxidoreductase. PMID- 6299339 TI - Possible role of sulphatide in the K+-activated phosphatase activity. AB - A microsomal fraction rich in (Na+ + K+)-ATPase has been isolated from the outer medulla of pig kidney. (Mg2+ + K+)-activated ouabain-sensitive phosphatase activity was studied in this preparation treated with arylsulphatase, an enzyme that specifically hydrolyzes ceramide galactose-3-sulphate. The activity of phosphatase was inactivated in proportion to the amount of sulphatide hydrolyzed. A maximum inactivation of ouabain-sensitive activity was obtained with 60% of the sulphatide content hydrolyzed. The inactivation caused by arylsulphatase was partially reversed by the sole addition of sulphatide. The evidence offered in this paper about sulphatide function in the sodium pump mechanism supports the idea that sulphatides are involved in the K+-activated phosphatase, a partial reaction of the (Na+ + K+)-ATPase. PMID- 6299340 TI - Temperature- and structure-dependent interaction of pyrethroids with the sodium channels in frog node of Ranvier. AB - (1) The interaction of a series of pyrethroid insecticides with the Na+ channels in myelinated nerve fibres of the clawed frog, Xenopus laevis, was investigated using the voltage clamp technique. (2) Out of 11 pyrethroids 9 insecticidally active compounds induce a slowly decaying Na+ tail current on termination of a step depolarization, whereas the Na+ current during depolarization was hardly affected. These tail currents are most readily explained by a selective reduction of the rate of closing of the activation gate in a fraction of the Na+ channels that have opened during depolarization. (3) The rate of decay of the Na+ tail current varies considerably with pyrethroid structure. After alpha-cyano pyrethroids the decay is at least one order of magnitude slower than after non cyano pyrethroids. The decay always follows a single-exponential time course and is reversibly slowed when the temperature is lowered from 25 to 0 degrees C. Arrhenius plots in this temperature range are linear. (4) These results indicate that the relaxation of the activation gate in pyrethroid-affected Na+ channels is governed by an apparent first order, unimolecular process and that the rate of relaxation is limited by a single energy barrier. Application of transition state theory shows that after alpha-cyano pyrethroids this energy barrier is 9.6 kJ/mol higher than after non-cyano pyrethroids. (5) Differences in rate of decay of the Na+ tail current account for the reported differences in repetitive nerve activity induced by various pyrethroids. In addition, the effect of temperature on the rate of decay explains the increase in repetitive activity with cooling. PMID- 6299341 TI - A spin-label study of plasma membranes of adrenal chromaffin cells. AB - Chromaffin-cell membranes were labeled with two nitroxide spin labels, one probing the interior of the membrane and one probing the interfacial region. Both spin labels indicate that the membrane undergoes a phase transition at about 26 degrees C. An Arrhenius plot of acetylcholinesterase activity exhibits a discontinuity at 26 degrees C, consistent with the existence of a phase transition at that temperature. Acetylcholine, which stimulates chromaffin cells to secrete catecholamines, and hexamethonium, a cholinergic blocker, do not affect the rotational correlation times of the spin labels. These results argue that cholinergic stimulation does not affect the fluidity of the chromaffin-cell membrane. PMID- 6299342 TI - Regulation of canine heart sarcolemmal Ca2+-pumping ATPase by cyclic GMP. AB - Cyclic nucleotide modulation of the sarcoplasmic reticulum calcium pump has been recognized for some time. Little is known, however, of cyclic nucleotide effects on the sarcolemmal Ca2+-pump. In sarcolemmal vesicles prepared from ventricular muscle by a recent technique (Jones, L.R., Maddock, S.W. and Besch, H.R. (1980) J. Biol. Chem. 255, 9971-9980) we have demonstrated via Millipore filtration that 10(-8) M and 10(-9) M cyclic GMP depressed the rate of ATP- and Mg2+-dependent 45Ca2+ uptake by 34% and 52%, respectively. Only at millimolar levels did cyclic AMP have any effect and the respective 5'-nucleotides had no effect at all. Parallel measurement of the associated (Ca2+ + Mg2+)-ATPase in the presence of either cyclic or 5'-nucleotides, however, revealed no concomitant depression in ATP hydrolysis. In another series of experiments, the cyclic GMP effect on 45Ca2+ uptake was associated with a significant decrease in the pump Vmax, and at the most effective concentration of cyclic GMP increased the apparent Km for Ca2+. These results suggest that cyclic GMP may depress ventricular Ca2+ efflux by decreasing the enzyme turnover and to a limited extent, decreasing pump affinity for Ca2+. This supports a hypothesis whereby cyclic GMP might modulate both local biochemical and electrophysiological events by an effect on a discrete, regional pool of intracellular Ca2+. PMID- 6299343 TI - Mechanism of passive Ca2+ permeability of vesicular sarcolemmal preparations from rat hearts. AB - Vesicular sarcolemmal preparations isolated from rat hearts were characterized by high total ATPase (4.32 +/- 0.57 mumol/min per mg), adenylate cyclase (121 +/- 11 pmol/min per mg) and creatine kinase (1.73 +/- 0.35 mumol/min per mg) activities as well as Na-Ca exchange specific to sodium. ATPase activity was inhibited with digitoxigenin by 50-70% and was not changed by ouabain, ionophore A23187 or oligomycin. Sarcolemmal vesicles bound [3H]digitoxigenin and [3H]ouabain in isotonic medium in the presence of Pi and Mg2+. The number of binding sites for hydrophobic digitoxigenin (N = 237 pmol/mg) was several-times higher than that for hydrophilic ouabain (N = 32.7 pmol/mg). These data show that sarcolemmal preparations were not significantly contaminated by mitochondria and sarcoplasmic reticulum and consisted mostly of inside-out vesicles. Incubation of these vesicles with 45Ca2+ (0.5-10 mM) led to penetration of the latter into the vesicles with the following binding characteristics: number of binding sites (N = 20.5 +/- 4.6 nmol/mg, Kd approximately equal to 2.0 mM). Ca2+ binding to the inner surface of vesicles was proved by the following facts: (1) Ca2+ ionophore A23187 increased slightly total intravesicular Ca2+ content but markedly accelerated Ca2+ efflux along its concentration gradient; (2) gramicidin and osmotic shock showed a similar accelerating effect. Ca2+ efflux from the vesicles along its concentration gradient ([Ca2+]i/[Ca2+]e = 2.0 mM/0.1 microM) was inhibited by Mn2+, Co2+, and verapamil when they acted inside the vesicles. The rate of Ca2+ efflux was hyperbolically dependent on intravesicular Ca2+ concentration (Km approximately equal to 2.9 mM). These data reveal that Ca2+ efflux from sarcolemmal vesicles is controlled by Ca2+ binding to the sarcolemmal membrane. Ca2+ efflux from the vesicles was stimulated 1.7--times after incubation of vesicles with 0.2 mM MgATP or MgADP and 15-times after treatment with 0.2 mM adenylyl beta, gamma-imidodiphosphate. Enhancement in the rate of Ca2+ efflux correlated with the increase in the intravesicular Ca2+ content. ATP stimulated Ca2+ efflux was suppressed by verapamil and was nonmonotonically dependent upon the transmembrane potential created by the K+ concentration gradient in the presence of valinomycin, Ca2+ efflux being slower at extreme values of membrane potential (+/- 80 mV). PMID- 6299344 TI - Perturbations of liver plasma membranes induced by Ca2+ are detected using a fatty acid spin label and adenylate cyclase as membrane probes. AB - Ca2+ decreased the lipid fluidity of rat liver plasma membranes labeled with 5 nitroxide stearate, I(12,3), as indicated by the order parameter (S). These effects form a reversible, saturable process with an association constant of 1 x 10(3) M-1. Arrhenius-type plots of S indicated that the lipid phase separation, present in the external leaflet of native membranes between 28 and 19 degrees C, is perturbed by mM Ca2+ such that the high temperature onset is elevated to 32-34 degrees C. Fluoride-stimulated adenylate cyclase was similarly inhibited by Ca2+ (ID50 = 1 mM) for the enzyme in membrane-bound or solubilized states. The glucagon-stimulated activity was more sensitive to Ca2+ inhibition with an ID50 of 0.2 mM. These inhibitory effects are due neither to perturbations of glucagon binding to its receptor nor to fluidity changes, but are instead attributed to direct Ca2+-enzyme interactions. Such binding desensitizes the enzyme to fluidity alterations induced by temperature elevation or benzyl alcohol addition. With Ca2+, Arrhenius plots of glucagon-stimulated activity indicated breaks at 32 and 16 degrees C, whereas those of fluoride-stimulated activity showed one break at 17 degrees C. Without Ca2+, Arrhenius plots exhibited one break at 28 degrees C for glucagon-stimulated activity, whereas fluoride-stimulated plots were linear. We propose that Ca2+ achieves these effects through asymmetric perturbations of the membrane lipid structure. PMID- 6299345 TI - Approximate dimensions of membrane lesions produced by streptolysin S and streptolysin O. AB - Membrane lesions produced by the streptococcal membranolysins streptolysin S and streptolysin O were investigated. Escape of labeled marker molecules of various sizes from resealed sheep erythrocyte ghosts treated with the toxins for 30 min allowed estimation of the sizes of the primary channels formed. Streptolysin S formed lesions ranging in size up to 45 A in diameter, and even high toxin concentrations did not result in larger channels. The lesions produced by streptolysin O exceeded 128 A in diameter. Kinetics experiments demonstrated that the primary streptolysin O lesions were formed rapidly (1-2 min), but release of marker molecules from streptolysin S-treated vesicles began only after a 5-15-min lag period. Label release from large unilamellar liposomes treated with streptolysin S suggested that membrane fluidity does not affect the size of the streptolysin S lesions. PMID- 6299346 TI - Calcium inhibition of the ATPase and phosphatase activities of (Na+ + K+)-ATPase. AB - In experiments performed at 37 degrees C, Ca2+ reversibly inhibits the Na+-and (Na+ + K+)-ATPase activities and the K+-dependent phosphatase activity of (Na+ + K+)-ATPase. With 3 mM ATP, the Na+-ATPase was less sensitive to CaCl2 than the (Na+ + K+)-ATPase activity. With 0.02 mM ATP, the Na+-ATPase and the (Na+ + K+) ATPase activities were similarly inhibited by CaCl2. The K0.5 for Ca2+ as (Na+ + K+)-ATPase inhibitor depended on the total MgCl2 and ATP concentrations. This Ca2+ inhibition could be a consequence of Ca2+-Mg2+ competition, Ca . ATP-Mg . ATP competition or a combination of both mechanisms. In the presence of Na+ and Mg2+, Ca2+ inhibited the K+-dependent dephosphorylation of the phosphoenzyme formed from ATP, had no effect on the dephosphorylation in the absence of K+ and inhibited the rephosphorylation of the enzyme. In addition, the steady-state levels of phosphoenzyme were reduced in the presence both of NaCl and of NaCl plus KCl. With 3 mM ATP, Ca2+ alone sustained no more than 2% of the (Na+ + K+) ATPase activity and about 23% of the Na+-ATPase activity observed with Mg2+ and no Ca2+. With 0.003 mM ATP, Ca2+ was able to maintain about 40% of the (Na+ + K+) ATPase activity and 27% of the Na+-ATPase activity seen in the presence of Mg2+ alone. However, the E2(K)-E1K conformational change did not seem to be affected. Ca2+ inhibition of the K+-dependent rho-nitrophenylphosphatase activity of the (Na+ + K+)-ATPase followed competition kinetics between Ca2+ and Mg2+. In the presence of 10 mM NaCl and 0.75 mM KCl, the fractional inhibition of the K+ dependent rho-nitrophenylphosphatase activity as a function of Ca2+ concentration was the same with and without ATP, suggesting that Ca2+ indeed plays the important role in this process. In the absence of Mg2+, Ca2+ was unable to sustain any detectable ouabain-sensitive phosphatase activity, either with rho nitrophenylphosphate or with acetyl phosphate as substrate. PMID- 6299347 TI - Intestinal brush border hydrolase topography. Effects of vitamin D-3 and filipin. AB - Intestinal brush borders were isolated from vitamin D-3-treated and vitamin D deficient chicks, and protein topography in the paired preparations assessed by the enzymatic release of four marker hydrolases. Exposure of the brush borders to the protease bromelain resulted in soluble levels of alkaline phosphatase, leucine aminopeptidase, maltase, and sucrase activities from preparations of vitamin D-3-treated birds that were 42%, 75%, 64%, and 56%, respectively, of corresponding activities released in preparations from rachitic chicks. Analyses for recovery of enzyme activity revealed that bromelain treatment selectively inactivated 43% of the alkaline phosphatase activity of brush borders obtained from vitamin D-3-replete birds, and preferentially diminished recovered sucrase activity in preparations from vitamin D-deficient chicks. In additional experiments, brush borders isolated from rachitic birds were treated in vitro with the polyene antibiotic filipin or an equivalent volume of vehicle. Subsequent exposure of such preparations to bromelain resulted in little or no differences in levels of marker hydrolase specific activities released from filipin- or vehicle-treated brush borders. However, analyses of membrane-bound specific activities after treatment of brush border preparations with a range of filipin concentrations, revealed a biphasic inhibition of approx. 30% for both maltase and sucrase, relative to vehicle controls, and a smaller effect on alkaline phosphatase and leucine aminopeptidase. PMID- 6299348 TI - Ca2+ displacement by Polymyxin B from sarcolemma isolated by 'gas dissection' from cultured neonatal rat myocardial cells. AB - Amphiphilic, cationic Polymyxin B is shown to displace Ca2+ from 'gas dissected' cardiac sarcolemma in a dose-dependent, saturable fashion. The Ca2+ displacement is only partially reversible, 57% and 63%, in the presence of 1 mM or 10 mM Ca2+, respectively. Total Ca2+ displaced by a non-specific cationic probe, lanthanum (La3+), at maximal displacing concentration (1 mM) was 0.172 +/- 0.02 nmol/microgram membrane protein. At 0.1 mM, Polymyxin B displaced 42% of the total La3+-displaceable Ca2+ or 0.072 +/- 0.01 nmol/microgram protein. 5 mM Polymyxin displaced Ca2+ in amounts equal to those displaced by 1 mM La3+. Pretreatment of the membranes with neuraminidase (removal of sialic acid) and protease leads to a decrease in La3+-displaceable Ca2+ but to an increase in the fraction displaced by 0.1 mM Polymyxin from 42% to 54%. Phospholipase D (cabbage) treatment significantly increased the La3+-displaceable Ca2+ to 0.227 +/- 0.02 nmol/microgram protein (P less than 0.05), a gain of 0.055 nmol. All of this phospholipid specific increment in bound Ca2+ was displaced by 0.1 mM Polymyxin B. The results suggest that Polymyxin B will be useful as a probe for phospholipid Ca2+-binding sites in natural membranes. PMID- 6299349 TI - Modulation of cardiac glycoside inhibition of (Na+ + K+)-ATPase by membrane lipids. Difference between species. AB - The role of lipids in the modulation of the ouabain-sensitivity of membrane (Na+ + K+)-ATPase from different species has been studied using a reconstitution procedure which promotes lipid exchange during detergent depletion by Sephadex chromatography. Hybrid reconstitution of delipidated (Na+ + K+)-ATPase preparations from bovine brain into the lipids obtained from crab nerve enzyme preparations significantly reduces the sensitivity of the brain enzyme to inhibition by ouabain. Conversely, reconstitution of crab nerve enzyme into the lipids from bovine brain enzyme preparations increases the sensitivity of the crab enzyme to ouabain inhibition. These opposing effects demonstrate the role of membrane lipids in modulating the enzyme-inhibition relationship in preparations from these different species. PMID- 6299350 TI - Lipid and protein contributions to the membrane surface potential of vesicular stomatitis virus probed by a fluorescent pH indicator, 4-heptadecyl-7 hydroxycoumarin. AB - The surface potential of membranes of vesicular stomatitis virus and liposomes was determined by shift of ionization over a wide pH range of the membrane inserted fluorophore, 4-heptadecyl-7-hydroxycoumarin. Incorporation into sonicated vesicles of negatively charged phosphatidylserine markedly increased the surface potential of uncharged phosphatidylcholine, but no significant effect on surface potential was produced by polar but uncharged glucocerebroside incorporated in phosphatidylcholine vesicles. The membrane of vesicular stomatitis virus was found to have a moderately high surface potential. Contributing to this viral membrane surface potential were glycoprotein spikes and phospholipid headgroups as determined by lowered charge after treatment of intact virions with thermolysin to remove glycoprotein or phospholipase C to remove phospholipid headgroups. The role of viral glycoprotein was confirmed by demonstrating increased surface charge of vesicles reconstituted with both viral glycoprotein and lipids compared with vesicles reconstituted with viral lipids alone. An unexpected finding was the large contribution to surface potential of cholesterol present in viral membrane. Increasing cholesterol concentration in virions by interaction with cholesterol-complexed serum lipoproteins resulted in a marked decrease in surface potential, whereas 75% depletion of virion cholesterol by interaction with sphingomyelin-complexed serum lipoproteins resulted in a significant increase in virion membrane surface potential. Although removal of glycoprotein spikes or depletion of cholesterol causes reduction in infectivity of vesicular stomatitis virus, no direct correlation could be found between alteration in surface charge and infectivity. PMID- 6299351 TI - A carbene-generating photoaffinity probe for beta-adrenergic receptors. AB - A new radioiodinated (2.2 Ci/mumol) iodocyanopindolol derivative carrying a 4-(3 trifluoromethyldiazirino)benzoyl residue has been synthesized. The long wavelength absorption of the diazirine permits formation of the carbene by photolysis under very mild conditions. [125I]ICYP-diazirine binds with high affinity (Kd = 60 pM) to beta-receptors from turkey erythrocyte membranes. Upon irradiation, [125I]ICYP-diazirine is covalently incorporated in a Mr 40 000 protein. Stereoselective inhibition of photolabeling by the (-)enantiomers of alprenolol and isoproterenol indicated that the Mr 40 000 protein contains a beta adrenergic binding site. The yield of specific labeling was up to 8.2% of total beta-receptor binding sites. The Mr 40 000 protein photolabeled in the membrane could be solubilized at comparable yield with either digitonin or Triton X-100. Irradiation of digitonin-solubilized turkey erythrocyte membranes with [125I]ICYP diazirine resulted in specific labeling of two proteins with Mr 40 000 and 50 000. In guinea-pig lung membranes, at least five proteins were photolabeled, of which one (with approximately Mr 67 000) was labeled specifically. PMID- 6299353 TI - D-glucose transport by membrane vesicles from quiescent, serum-stimulated, and SV40-transformed mouse 3T3 cells. AB - Mixed membrane vesicle preparations from mouse embryo fibroblasts (Swiss 3T3) exhibited a facilitated diffusion transport system for D-glucose that showed many of the characteristics of the D-glucose transport system of whole cells: stereospecificity, counterflow, Michaelis-Menten kinetics with an apparent Km similar to that of whole cells, and sensitivity to inhibition by cytochalasin B. Comparison of the stereospecific D-glucose transport activities of membrane vesicles from quiescent, serum-stimulated, and SV40 virus-transformed 3T3 cells showed no significant differences in rates of D-glucose uptake or efflux. This is in contrast to whole cells; quiescent 3T3 cells transported 6-deoxy-D-glucose at a significantly lower rate than serum-stimulated or SV40-transformed cells. These results indicate that D-glucose transport in quiescent vs. actively growing cells is regulated by cellular factors that are not retained in membrane vesicle preparations. PMID- 6299352 TI - Specificity in the interaction of phospholipids and fatty acids with vesicle reconstituted cytochrome P-450. A spin label study. PMID- 6299354 TI - Isolation and characterization of plasma membranes from Friend erythroleukaemic cells. A study with sphingomyelinase C. AB - Plasma membranes have been prepared from Friend erythroleukaemic cells using a Dounce homogenization technique followed by differential and sucrose gradient centrifugations. (I) A plasma membrane fraction was obtained which showed a 20- to 30-fold enrichment in 5'-nucleotidase, alkaline phosphodiesterase I, alkaline phosphatase and in 32P-labeled (poly)phosphoinositides. About 1% of the total protein, 6-7% of phospholipid, 8-9% of cholesterol and 12-15% of each of the above markers were recovered in the plasma membrane fraction with an average yield of 15-20%. The plasma membrane was characterized by a high cholesterol to phospholipid molar ratio (0.626), a 2-fold enrichment in sphingomyelin and in phosphatidylserine as compared to the whole cell and by the complete absence of diphosphatidylglycerol. (2) When compared to the phospholipid composition of the mature mouse erythrocyte membrane, the plasma membrane of the Friend cell only differs by a higher phosphatidylcholine and a lower phosphatidylethanolamine content, whereas the levels of sphingomyelin and phosphatidylinositol plus phosphatidylserine are similar. (3) Friend cells were treated with sphingomyelinase C (S. aureus) under non-lytic conditions and subsequently submitted to subcellular fractionation. The results showed that the plasma membrane accounted for 38.5% of the total phospholipid, 64.1% of the total cholesterol and about 4.4% of the total protein content of Friend cells. (4) Sphingomyelin appeared to be asymmetrically distributed in the plasma membrane of Friend cells, with about 85% of this phospholipid being present in the outer monolayer. PMID- 6299355 TI - Sodium channel activity in brain membrane fractions isolated from rats of different ages. AB - The Na+ channel activity (tetrodotoxin sensitive 22Na+ flux induced by veratridine and/or anemone toxin II) was studied in two fractions of brain cell plasma membranes, named A and B, isolated by the method of Gray and Whittaker ((1962) J. Anat. 96, 79-87) from rats 5, 10, 30 and 60 days old. The 22Na+ flux was measured in membrane vesicles formed by the isolated membranes, in the absence of drugs (control), in the presence of veratridine, and in the presence of veratridine plus tetrodotoxin. Fraction A consists primarily of neuronal and glial membranes in rats of 5 and 10 days of age, while in the older rats this fraction becomes enriched in myelin. In Fraction A of 5-day-old and 10-day-old rats, veratridine (25 microM) increases the 22Na+ flux 2.4- and 1.6-fold, respectively, and the increment continues to diminish with age, until it becomes negligible in the 60-day-old rats. Fraction B consists of synaptosomes and membrane vesicles, and at the four ages studied veratridine (25 microM) causes an increment of the 22Na+ flux of about 2.5-fold. Fractions A and B from 10-day-old rats, and Fraction B from 60-day-old rats, which are sensitive to veratridine, also respond to anemone toxin II. When veratridine is used in presence of anemone toxin II (0.5 microM), the K0.5 for veratridine is diminished and the maximum 22Na+ flux is increased. The increments of 22Na+ flux caused by veratridine and/or anemone toxin II in Fractions A and B are blocked by tetrodotoxin (K0.5 approx. 5 nM). Fraction A from 60-day-old rats could be subfractionated by osmotic shock and sucrose gradient centrifugation to obtain three subfractions, two of which are enriched in axolemma and display Na+ channel activity. The other subfraction is enriched in myelin and shows no Na+ channel activity. The plasma membrane preparations from young rats (up to 10 days) are devoid of myelin and are useful for studies of Na+ channel activity. PMID- 6299356 TI - Expression of integrated viral DNA sequences outside the transforming region in eight adenovirus-transformed cell lines. AB - The expression of early and intermediate-early viral regions in eight adenovirus type 5 transformed cell lines was analyzed by radioimmuno-inhibition and RNA-DNA hybridization techniques. Details on the arrangement of the integrated viral DNA sequences in these cell lines have already been published (Visser, L., Wassenaar, A.D.C., Van Maarschalkerweerd, M.W. and Rozijn, T.H. (1981) J. Virol, 39, 684 693). In all cell lines tested, proteins encoded by the transforming region E1 are present. Dependent on the viral DNA content, additional early regions are expressed in most cell lines. In two of the cell lines polypeptides related to the adenoviral terminal protein, encoded by the recently described region E2b, could be detected. The viral DNA sequence encoding the body of the terminal protein mRNA is probably integrated intact, but the promoter region and at least some of the leaders are lacking in these cell lines. PMID- 6299357 TI - Characterization of a 45-kDa polypeptide as the precursor of subunit 1 of cytochrome c oxidase in Neurospora crassa. AB - In a previous paper (Van 't Sant, P., Mak, J.F.C. and Kroon, A.M. (1981) Eur. J. Biochem. 121, 21-26) we showed the existence of three elongated precursor proteins (45, 36 and 25 kDa) of mitochondrial translation products in Neurospora crassa. We presented some indications that the largest precursor could be related to subunit 1 of cytochrome c oxidase. Here we present conclusive evidence that the 45-kDa polypeptide is indeed this precursor by demonstrating that an immunodetectable 45-kDa polypeptide displays the same behaviour as the labeled 45 kDa precursor; both accumulate after long incubation with cycloheximide or by decreasing the temperature and both are not tightly membrane bound. Moreover the antibody against subunit 1 of cytochrome c oxidase also recognizes, in immunoadsorption experiments, besides subunit 1, the 45-kDa polypeptide accumulated by cycloheximide incubation. Furthermore, we developed a small scale purification of antibodies against subunit 1 of cytochrome c oxidase. By means of these purified antibodies it is demonstrated that the 45-kDa polypeptide and subunit 1 have corresponding antigenic determinants. Under the various conditions tested, all three precursors are less firmly membrane-bound than the mature subunits. Finally, it is observed that in short incubations in vivo, chloramphenicol inhibits the processing of the mitochondrially synthesized precursors, under conditions where mitochondrial translation is only partially inhibited. PMID- 6299358 TI - Characterization of the DNA-cellulose-binding proteins from Escherichia coli K 12. AB - The various [35S]DNA-binding proteins present in lysates of Escherichia coli K 12 cells have been analyzed by means of two-dimensional SDS-polyacrylamide gel electrophoresis. The proteins were isolated by the DNA-cellulose technique and eluted by increasing concentrations of NaCl (0.15, 0.4, 0.6 and 2 M). Only 2% of the total 35S radioactivity in the lysate became bound to the DNA-cellulose column. A total of 237 polypeptides were detected and the distribution among the salt eluates were 85, 83, 40 and 29 polypeptides, respectively. The 40 major polypeptides with regard to concentrations were also identified from gels stained with a protein-specific reagent. The polypeptides could be divided into two main groups according to pI values, namely, acidic polypeptides (total number, 174) and basic polypeptides (total number, 63). The ratio between acidic and basic polypeptides decreased with increasing salt concentrations in the eluates. The majority of the basic polypeptides had molecular weights in the range 10 000-30 000, whereas the acidic polypeptides had molecular weights from 10 000 to 165 000. PMID- 6299359 TI - Evidence for extensive methylation of ribosomal RNA genes in a rat XC cell line. AB - XC cell line was established from a rat fibrosarcoma induced by avian sarcoma virus. Analysis of ribosomal RNA genes in the DNA of this cell line indicated that there was a 3- to 4-fold increase in the number of the rDNA repeating units. [methyl-14C]Methionine labeling experiments as well as Southern blot analysis of the restriction endonuclease fragments showed extensive methylation of the cytosine residues in the rDNA. Further analysis by HpaII, HhaI and MspI suggested that the entire rDNA repeat unit was uniformly methylated in the recognition sequence for these enzymes. Parallel experiments with two other rat cell lines (rat embryo fibroblasts and adenovirus-transformed rat embryo cells) indicated no evidence of methylation in the rDNA. PMID- 6299360 TI - Triiodothyronine nuclear receptor. Role of histones and DNA in hormone binding. AB - The triiodothyronine (T3) nuclear receptor was previously shown to lose rapidly its high affinity hormone-binding property after a partial purification from the nuclear extract. It was then found that histones + DNA added to the incubation medium with labeled T3 could restore, at least in part, the high affinity T3 binding. We now demonstrate that DNA alone increases the high affinity T3 binding site concentration moderately, and only at low ionic strength where it can bind to the receptor. Total histones and all histone fractions studied (total core histones, F2a, H2B, H3, H4, H1) specifically increase, at low concentrations, the level of T3 binding; but higher concentrations of some individualized histones, particularly arginine-rich histones, have an inhibitory effect. DNA, or several other polynucleotides, in the presence of histones increase the stimulating histone effect and reverse the inhibitory effect into a true activation. Histones increase the number of T3 binding sites but decrease the affinity for T3; addition of DNA restores the high affinity for T3 and stabilizes the T3-receptor complexes. Thus, some of the histone molecules could play a role in the maintenance of the T3 binding site, but multiple interactions between histones or with DNA seem necessary to impair the negative effect exerted by other parts of the histone molecules. Whether these positive and negative effects of histones on the T3 binding site are of biological relevance in the regulation of T3 binding to its receptor remains to be determined. PMID- 6299361 TI - Characterization of phosphoprotein phosphatases and phosphorylase phosphatase from yeast. AB - Three peaks of protein phosphatase (phosphoprotein phosphohydrolase, EC 3.1.3.16) activity (fractions a, b and c) acting on muscle phosphorylase (1,4-alpha-D glucan:orthophosphate alpha-D-glucosyltransferase, EC 2.4.1.1) were separated by DEAE-cellulose chromatography of yeast extracts. In contrast to fractions a and b, only fraction c was able to liberate phosphate from 32P-labelled inactivated yeast phosphorylase. The activity of fraction c on both substrates was totally dependent on the presence of bivalent metal ions (Mg2+, Mn2+), and was activated by Mg . ATP. Following freezing in the presence of mercaptoethanol, fractions a and b were also able to dephosphorylate yeast phosphorylase. Rabbit muscle phosphoprotein phosphatase inhibitors 1 and 2 showed that yeast phosphatases acting on muscle phosphorylase were inhibited by inhibitor 2 but not by inhibitor 1. The action of fraction c on yeast phosphorylase was not inhibited by either inhibitor. The native yeast phosphorylase phosphatase (EC 3.1.3.17) was purified 8000-fold by ion-exchange chromatography, casein-Sepharose chromatography and Sephadex G-200 gel filtration. The purified enzyme was unable to dephosphorylate rabbit muscle phosphorylase a, but acted on casein phosphate (Km 3.3 mg/ml). Molecular weight was estimated to be 78 000 and pH optimum 6.5-7.5. Activity of the enzyme was dependent on bivalent metal ions (Mg2+, Mn2+) and was inhibited by fluoride (Ki 20 mM) and succinate (Ki 10 mM). PMID- 6299362 TI - Properties of the cyclic GMP phosphodiesterase from rat lung. Inhibition by unsaturated fatty acids. AB - Catalytic and regulatory properties of the major form of cyclic GMP phosphodiesterase (3':5'-cyclic-GMP 5'-nucleotidohydrolase, EC 3.1.4.35) from rat lung were studied. The enzyme partially purified by a DEAE-Sepharose chromatography displayed a much higher affinity toward cyclic GMP than toward cyclic AMP, the apparent Km values being 5.7 microM and 482 microM for the guanylic and the adenylic cyclic nucleotide, respectively. In contrast, the V value for cyclic AMP was about 3-times higher than the V value for cyclic GMP. Linear double reciprocal plots of initial velocity were observed with each cyclic nucleotide. From 10(-8) to 3.3 X 10(-6) M, cyclic GMP did not change the hydrolysis of 1 or 10 microM cyclic [3H]AMP, while it became inhibitory at higher concentrations. In contrast with a calmodulin-sensitive phosphodiesterase prepared from rat brain, the lung enzyme was not stimulated by a heat-stable Ca2+ dependent factor from rat lung or by rat brain calmodulin or by lipids including fatty acids and lysophosphatidylcholine. Various unsaturated 18- and 20-carbon fatty acids inhibited at varying degrees the cyclic GMP phosphodiesterase from rat lung. The inhibitory potency increased with the number of double bonds in the hydrocarbon chain. In contrast, the methyl esters of the unsaturated fatty acids and the saturated fatty acids of variable hydrocarbon chain lengths had no appreciable effects. A linear Hill plot of phosphodiesterase inhibition with a slope of unity was obtained with arachidonic acid up to 30 microM, suggesting only one type of inhibitory site. In this range of concentrations the inhibition was entirely reversible. Kinetics analysis demonstrated that up to 30 microM arachidonic acid was a purely competitive inhibitor with an apparent Ki of 20 microM. Over 30 microM, the Hill coefficient increased progressively, indicating the binding to other inhibitory sites, while the reversibility disappeared. PMID- 6299363 TI - The ionic strength dependence of the rate of a reaction between two large proteins with a dipole moment. PMID- 6299364 TI - Electron addition to the (FeO2) unit of oxyhaemoglobin Glycera. AB - Exposure of glassy solutions containing the monomeric fraction of the oxyhaemoglobin of the polychaete annelid Glycera dibranchiata to 60Co gamma-rays at 77 K resulted in electron addition to the (FeO2) moiety. The form of the g tensor components obtained from the ESR spectrum indicates that the spin-density on oxygen is much greater than that observed for similar paramagnetic centres formed in haemoglobin A or myoglobin. A major difference between these monomer haem units and normal haem units is that the distal histidine (E7 58) is replaced by leucine. We therefore postulate that the oxygen in the (FeO2)- units formed in haemoglobin A and myoglobin is hydrogen-bonded to the NH group of the distal histidine, whilst that of the (FeO2)- units in haemoglobin Glycera are not hydrogen-bonded. However, on annealing to approx. 160 K the spectrum changed irreversibly into one resembling those for (FeO2)- units in haemoglobin A and myoglobin. We postulate that this is caused by hydrogen-bonding to a water molecule in the haem pocket. Exposure of the polymeric fractions of haemoglobin Glycera to gamma-rays gave an (FeO2)- unit with an ESR spectrum remarkably similar to that obtained from oxymyoglobin. The X-ray structure of this protein is unknown but we suggest that our results could indicate the presence of a distal histidine in this material. PMID- 6299365 TI - Aluminum interaction with calmodulin. Evidence for altered structure and function from optical and enzymatic studies. AB - The interaction of aluminum ions with bovine brain calmodulin has been examined by fluorescence spectroscopy, circular dichroic spectrophotometry and equilibrium dialysis, and by the calmodulin-dependent activation of 3',5'-cyclic nucleotide phosphodiesterase. These experiments show that aluminum binds stoichiometrically and cooperatively to calmodulin. Binding of aluminum at a molar ratio of 2:1 to calmodulin suffices to induce a major structural change. Estimates from spectroscopic data indicate that the binding affinity for the first mol of aluminum bound to the protein is about one order of magnitude stronger than that of calcium to its comparable site. These estimates agree with a dissociation constant of 0.4 microM derived from equilibrium dialysis experiments. Interaction of aluminum with calmodulin induces a helix-coil transition and enhances the hydrophobic surface area much more than calcium does. A molar ratio of 4:1 for [aluminum]/[calmodulin] is sufficient to block completely the activity of the calcium-calmodulin-dependent phosphodiesterase. Highly hydrated aluminum ions apparently promote solvent-rich, disordered polypeptide regions in calmodulin which, in turn, profoundly influence the protein's flexibility. PMID- 6299366 TI - Cytosolic modulators of activities of microsomal enzymes of cholesterol biosynthesis. Effects of Acyl-CoA inhibition and cytosolic Z-protein. AB - Physiological concentrations of long-chain fatty acyl-CoAs have now been shown to inhibit microsomal methyl sterol oxidase. Acyl-CoA inhibition of hydroxymethylglutaryl-CoA reductase as well as methyl sterol oxidase can be either prevented or reversed by the addition of purified Z-protein (fatty acid binding protein). Concomitantly, Z-protein addition decreases the extent of binding of radioactively labeled oleoyl-CoA to microsomal membranes. Free heme also inhibits hydroxymethylglutaryl-CoA reductase, and Z-protein reverses the extent of observed inhibition by binding heme analogous to the effect observed with acyl-CoAs. Similarly, Z-protein reverses substrate inhibition of acyl CoA:cholesterol acyltransferase at high concentrations of acyl-CoA substrate. All these observations are consistent with the suggestion that, by binding acyl-CoAs and other enzyme effectors such as free heme, Z-protein modulates the effects of fluctuations of concentrations of major cellular metabolites. Furthermore, because the concentration of Z-protein is very low in rapidly growing hepatomas, such tumors may be very poorly buffered against the effects of acyl-CoAs, free fatty acids, heme and other effectors that may vary markedly by either altered metabolism or release of metabolites from necrotic tumor tissue. PMID- 6299367 TI - Formation of catechol estrogens by intestinal bacterial demethylation of 2 methoxyestrone. AB - The intestinal bacterial metabolism of 2-methoxyestrone was studied by incubation in the isolated coecum from rats. Following isolation of estrogens by a combination of ion-exchange and ligand-exchange chromatography, the metabolites were identified by gas chromatography-mass spectrometry. The two main reactions were oxidoreduction at C-17 and extensive demethylation at C-2. Thus, the demethylation of 2-methoxyestrogens known to occur in vivo may be due to the action of microbial enzymes. The study also shows that the intestinal microflora is capable of converting biologically inactive into active steroid hormones. PMID- 6299368 TI - Iodination of arachidonic acid mediated by eosinophil peroxidase, myeloperoxidase and lactoperoxidase. Identification and comparison of products. AB - Arachidonic acid undergoes iodination in the presence of hydrogen peroxide, iodide, and either eosinophil peroxidase, myeloperoxidase or lactoperoxidase. The profile of products generated by each of the three peroxidases is similar as determined by reversed-phase high-performance liquid chromatography. Structural analysis of the products indicate that: 1, each of the four double bonds in arachidonic acid is susceptible to iodination; 2, arachidonic acid can be multiply iodinated; and 3, the carboxylate moiety does not participate in the formation of all products. The isomeric composition of the isolated products indicates that peroxidase-mediated iodination of arachidonate is not stereoselective. PMID- 6299369 TI - Separation of eucaryotic ribosomal proteins by high-performance liquid chromatography using a new cation-exchange resin and LiCl elution in 4 M urea. PMID- 6299370 TI - Purification of equine gonadotropins and comparative study of their acid dissociation and receptor-binding specificity. PMID- 6299371 TI - Induction of phosphoribomutase in Bacillus cereus growing on nucleosides. AB - In this paper we show that phosphoribomutase is induced in Bacillus cereus by the same metabolizable purine and pyrimidine ribonucleosides previously shown to induce the purine nucleoside phosphorylase (Tozzi, M.G., Sgarrella, F. and Ipata, P.L. (1981) Biochim. Biophys. Acta 678, 460-466). The mutase allows ribose 1 phosphate formed from nucleosides to be utilized by the cell through the pentose cycle, upon transformation to ribose 5-phosphate. The equilibrium constant of the mutase reaction is towards ribose-5-phosphate formation. The coordinate induction of the two enzymes completes the picture of the molecular events leading to the utilization of the sugar moiety of purine nucleotides and nucleosides as an energy source (Mura. U., Sgarrella, F. and Ipata, P.L. (1978) J. Biol. Chem. 253, 7905-7909). PMID- 6299372 TI - Forskolin stimulation of acid and pepsinogen secretion by gastric glands. AB - Isolated gastric glands were used to investigate the action of forskolin, a novel diterpene extracted from the Indian plant Coleus forskohlii. Forskolin was found to stimulate both acid formation and pepsinogen secretion. The stimulation was rapid, reversible and dose dependent with an ED50 of approx. 1 microM. The efficacy of forskolin was similar to that of more commonly used secretagogues, e.g. histamine, carbachol, cyclic AMP derivatives. The responses to forskolin were not inhibited by any of the receptor-specific antagonists tested. Forskolin activated adenyl cyclase in gland homogenates over a dose range similar to that required for stimulation of secretory activity. Forskolin was found to be more effective in activating adenyl cyclase than histamine, isoproterenol or NaF. Treatment of gastric glands with forskolin resulted in a 100-fold increase in tissue cAMP levels, supporting the idea that forskolin activates adenyl cyclase in the intact cell. The results are interpreted to show that forskolin stimulation of gastric secretions is due to activation of adenyl cyclase with a consequent increase in tissue cAMP. PMID- 6299373 TI - Possible identity of a membrane-bound with a soluble cyclic AMP-independent erythrocyte protein kinase that phosphorylates spectrin. AB - A soluble casein kinase isolated and purified to homogeneity from the human erythrocyte cytosol by phosphocellulose and Sephadex G-200 chromatographies is indistinguishable from the membrane-bound casein (spectrin) kinase according to physical and site-specificity criteria. The soluble enzyme shows an Mr of about 30000 by gel filtration and comigrates with the purified membrane spectrin kinase as a single polypeptide of 32000 Da on sodium dodecyl sulfate polyacrylamide gels. The soluble kinase phosphorylates spectrin in situ in spectrin kinase depleted ghosts and catalyzes the in vitro phosphorylation of partially dephosphorylated spectrin with saturation kinetics identical to those displayed by the membrane spectrin kinase. When component 2 of spectrin that had been phosphorylated with [gamma-32P]ATP by either the soluble or the membrane kinases was subjected to limited proteolysis, the same 21500 Da papain-generated phosphopeptide was found to have been produced by the two enzymes. The same 21500 Da phosphopeptide was identified after papain digestion of spectrin isolated from intact cells that had been incubated with 32Pi. However, this particular peptide was not labeled in spectrin that had been phosphorylated in vitro by the catalytic subunit of cyclic AMP-dependent protein kinase. Identical phosphopeptide patterns were obtained by gel filtration and two-dimensional peptide maps of trypsin-cleaved component 2 of spectrin that had been labeled in situ, in intact ghosts or in spectrin kinase-depleted ghosts supplemented with the soluble kinase. These findings indicate a possible identity of the soluble with the membrane-bound casein (spectrin) kinase. PMID- 6299374 TI - Hydroxyl free-radical spin-adduct in rat brain synaptosomes. Observations on the reduction of the nitroxide. AB - Understanding the prevalence and action of hydroxyl free-radicals, damaging species in biological systems, is improved by spin-trapping techniques. The hydroxyl free-radical rapidly adds to the spin-trap 5,5-dimethyl-1-pyrroline-1 oxide (DMPO) to form a relatively stable nitroxyl free-radical spin-adduct (DMPO OH), but it was found that DMPO-OH becomes diamagnetic, presumably by chemical reduction, when it is added to rat brain synaptosomes. DMPO-OH reduction by synaptosomes was a time-dependent process, starting immediately and continuing up to 20 min or more, when almost all the spin-adduct is reduced. During synaptosomal reduction, the electron paramagnetic resonance (EPR) high field line of DMPO-OH is broadened indicating restrictions on the motion of the nitroxide. This was not due to simple absorption of DMPO-OH to proteins. Experiments with the spin-adduct in glycerol at low temperatures showed that DMPO-OH experienced, during synaptosomal reduction, restricted motion having a correlation time of about 3.3 X 10(-9)s. This value would be expected if DMPO-OH was near the polar region of the hydrocarbon layer of a membrane bilayer. Synaptosomal membrane transport of DMPO-OH was tested by competition studies, since it was presumed that this nitroxide was transported and then reduced inside the synaptosomes. Several compounds tested, including amino acids, neurotransmitters, food dyes, and many others, that may have acted competitively in transport were not effective in slowing down reduction of DMPO-OH. However, it was found that azide, cyanide and heating of the synaptosomes markedly decreased the rate of synaptosomal reduction of DMPO-OH. Azide at 50 microM manifested one-half of its maximal inhibitory effect. Experiments with 1-octanol to obtain a measure of the membrane partition of DMPO-OH demonstrated that the octanol/water distribution was 1.83. Experiments with glass microfibre filtration to determine synaptosomal binding revealed that DMPO, as well as apparently DMPO-OH, were not specifically retained by synaptosomes. All of these results combined with past work on nitroxide reduction by biological systems, suggest that DMPO-OH reduction by synaptosomes is due to its interaction with and reduction by electron transport carriers of mitochondria within the synaptosomes. PMID- 6299375 TI - Effects of cyclic nucleotides on ornithine decarboxylase activity in mammary gland explants from mid-pregnant mice. AB - Dibutyryl cAMP and prolactin stimulated ornithine decarboxylase activity in mouse mammary gland explants which had been preincubated with insulin and cortisol for 1 day; maximally stimulatory concentrations of dibutyryl cAMP and prolactin produced a response which was greater than the sum of the responses of prolactin and dibutyryl cAMP when tested alone. 8-Bromo-cGMP inhibited ornithine decarboxylase activity whereas other derivatives of cyclic nucleotides were without effect. Cortisol concentrations were found to be important for optimizing the dibutyryl cAMP and prolactin responses. Optimal prolactin responses were obtained with cortisol concentrations greater than 10(-7) M, whereas optimal dibutyryl cAMP responses were observed with cortisol concentrations less than 10( 7) M. Despite the differing optimal cortisol concentrations for the prolactin and dibutyryl cAMP responses, it is concluded that prolactin and dibutyryl cAMP probably stimulate ornithine decarboxylase activity in the mammary gland via the same mechanism. PMID- 6299376 TI - Intracellular distribution of fumarase in rat skeletal muscle. AB - The distribution of fumarase activity between the mitochondrial and cytoplasmic compartments of rat skeletal muscle was studied using the method of Fatania and Dalziel (Biochim. Biophys. Acta 631 (1980) 11-19), fractional extraction technique and a method based on the calculation of mitochondrial protein content in the tissue and on the determination of fumarase activity both in the tissue homogenate and in the isolated mitochondria. We found 10%, 5% and 0% of the total fumarase activity in the cytoplasm using these methods, respectively. The results suggest that no more than 10% of the total fumarase activity is present in the cytosolic fraction of rat skeletal muscle. The metabolic consequences of such distribution of fumarase in skeletal muscle are discussed. PMID- 6299377 TI - Characterization of beta-adrenergic receptor subtypes in androgen-induced mouse kidney hypertrophy using a new high-affinity ligand, [125I]iodocyanopindolol. AB - In fully developed androgen-induced hypertrophy of female mouse kidney, beta adrenergic receptors per unit membrane protein were increased approx. 2.5-fold, as measured by the binding of [125I]iodocyanopindolol, with no change in apparent dissociation constants (Kd range 20-25 pM). Membrane protein relative to total kidney protein, Na+/K+-dependent ATPase (EC 3.6.1.3) and 5'-nucleotidase (EC 3.1.3.5) activities and cholesterol content per unit membrane protein did not differ significantly in preparations from control and treated animals. The binding of iodocyanopindolol to kidney membranes was characterized with respect to association and dissociation kinetics, and also in regard to the less-specific contributions of other major catecholamine or indolamine receptors, using mixtures of the corresponding specific competitors. beta 1-selective drugs, practolol and metoprolol, and beta 2-selective agents, IPS-339 and zinterol, were competed with iodocyanopindolol to assess the receptor type specificity, and the ensuing binding profiles were dissected by a nonlinear regression analysis as described by Munson, P.J. and Rodbard, D. (Anal. Biochem. (1982) 107, 220-239). Most of the androgen-induced beta-adrenergic receptors had the binding properties corresponding to beta 2-subtype. No consistent increase in the density of beta 1 adrenergic receptors could be shown. PMID- 6299378 TI - Effects of forskolin on adenylate cyclase, cyclic AMP, protein kinase and intermediary metabolism of the thyroid gland. AB - Forskolin (40 microM) stimulated adenylate cyclase activities of bovine thyroid plasma membranes without the addition of guanine nucleotides. GDP had little effect on the forskolin-stimulated adenylate cyclase activity while Gpp[NH]p (0.1 1.0 microM) decreased it. In the presence of TSH (10 mU/0.11), Gpp[NH]p no longer caused inhibition. Forskolin did not affect phosphodiesterase activities of thyroid homogenates. Forskolin (10 microM) rapidly increased cAMP levels in bovine thyroid slices both in the absence and presence of a phosphodiesterase inhibitor. The effect of TSH (50 mU/ml) on cAMP levels was additive or greater than additive to that of forskolin. An initial 2-h incubation of slices with forskolin did not decrease their subsequent cAMP responses to either forskolin and/or TSH while similar treatment of slices with TSH induced desensitization of the cAMP response to TSH, but not to forskolin. Forskolin (10 microM) as well as TSH (50 mU/ml) activated cAMP-dependent protein kinase of slices in the absence of a phosphodiesterase inhibitor. Although forskolin activated the adenylate cyclase cAMP system, it did not stimulate iodide organification or glucose oxidation, effects which have been attributed to cAMP. In fact, forskolin inhibited these parameters and 32P incorporation into phospholipids as well as their stimulation by TSH. These results indicate that an increase in cAMP levels and cAMP-dependent protein kinase activity in thyroid slices may not necessarily reproduce the effects of TSH on the thyroid. PMID- 6299379 TI - Interaction of thyroid hormones and cyclic AMP in the stimulation of hepatic gluconeogenesis. PMID- 6299380 TI - Delineation of membrane-bound phosphatase activities in normal and leukemic thymocytes. PMID- 6299381 TI - Effect of neuraminidase on luteinizing hormone/chorionic gonadotrophin binding to the ovarian and testicular membranes from different mammalian species. AB - After treatment of the ovarian and testicular membranes from several mammalian species an elevation in the specific binding of human [125I]-labelled CG could be observed. With the assumption that this effect is due to sialic acid-masked receptors, the presence of such receptors seem to be a common property of most mammalian gonads. An interesting observation was the abnormally high hormone binding capacity of the Syrian hamster ovary, as compared to other hamster species, and the lack of a neuraminidase effect in the ovary of the Syrian hamster. PMID- 6299382 TI - The effect of spin traps on phenylhydrazine-induced haemolysis. AB - Spin-trapping agents have been used to study the involvement of free radicals in phenylhydrazine-induced haemolysis. Spin traps were found to decrease the rate of oxygen uptake and the rate of haemoglobin oxidation in the reaction of phenylhydrazine with oxyhaemoglobin. Spin traps were also found to inhibit haemolysis and lipid peroxidation in phenylhydrazine-treated fresh human erythrocytes, with a concomitant production of phenyl radical spin adducts. Lipophilic spin traps were found to be more effective inhibitors of haemolysis than their hydrophilic analogues. PMID- 6299384 TI - Isolation and characterization of phosphoprotein pp 105 from simian virus 40 transformed mouse fibroblasts. AB - Investigation of the cellular distribution of a 105 kDa phosphoprotein (pp 105) in transformed mouse fibroblasts, showed that only a minor amount was located on the surface of logarithmically grown suspension cells. More than 90% of total pp 105 was contained in the cytosolic fraction representing about 0.2% of total cytosolic proteins. Surface and cytosolic pp 105 had identical phosphopeptide patterns. Cytosolic pp 105 was highly purified by ammonium sulfate precipitation followed by three chromatographic steps and gel electrophoresis. The purified pp 105 was capable of weak autophosphorylation. In the stationary growth phase of suspension cells, the amount of pp 105 detectable by endogenous phosphorylation was only 10-15% of that observed during logarithmic growth. pp 105 was also detected in normal mouse tissue and its distribution determined. PMID- 6299383 TI - Properties and subcellular localization of adenosine diphosphatase in arterial smooth muscle cells in culture. AB - The properties and subcellular localization of adenosine diphosphatase (ADPase) activity in smooth muscle cells cultured from pig aortas have been investigated. The pH optimum of ADPase activity was 7.3 and the apparent Km for ADP was 10.3 microM. ADPase activity was inhibited completely by EDTA and was restored by the addition of divalent cations. The enzyme activity was not inhibited by 2 glycerophosphate, a substrate for non-specific phosphatases, nor by levamisole, a specific inhibitor of alkaline phosphatase. Smooth muscle cells were homogenized and a post-nuclear supernatant was applied to a sucrose density gradient in a Beaufay automatic zonal rotor. The distribution of ADPase activity in the density gradient was similar to that of 5'-nucleotidase activity, a marker enzyme for the plasma membrane, and distinct from the distributions of the marker enzymes for the other organelles. When the cells were homogenized in the presence of digitonin, an agent which binds to cholesterol and increases the equilibrium density of the plasma membrane, the modal equilibrium densities of ADPase activity and of 5'-nucleotidase activity were increased to similar extents, thus confirming the plasma membrane localization of ADPase activity. PMID- 6299385 TI - Cyclic AMP and tumor promoters cause differential induction of ornithine decarboxylase and accumulation of putrescine in Chinese hamster ovary cells deficient in cyclic AMP-dependent protein kinase. PMID- 6299386 TI - Role of cyclic AMP-dependent protein kinase activation in regulating rat submandibular mucin secretion. AB - The extent of activation of rat submandibular gland cyclic AMP-dependent protein kinase (EC 2.7.1.37) was determined in vitro using dispersed cells to assess the involvement of this enzyme in submandibular mucin secretion. cAMP-dependent protein kinase activation, as determined by activity ratio method, was markedly increased following beta-adrenergic receptor activation. 0.5 M NaCl was required in the homogenization buffer for stabilization of the hormonally activated cAMP dependent protein kinase. A role for cAMP-dependent protein kinase activation in regulating mucin secretion was strongly suggested by the following: (1) the kinase activity ratio increased rapidly after beta-adrenergic receptor stimulation; (2) dose-response relationship of the kinase activation following beta-adrenergic receptor activation correlated with isoproterenol induced mucin release; (3) termination of beta-adrenergic mediated mucin secretion caused a rapid decrease in the kinase activity ratio; (4) dibutyryl cyclic AMP stimulation caused an increase in the kinase ratio; whereas (5) pure cholinergic and pure alpha-adrenergic receptor stimulation had no effect on endogenous kinase activity. Although cAMP-dependent protein kinase activation may not be the only regulator of mucin secretion, these data suggest an important regulatory role for this kinase activation during rat submandibular mucin release. PMID- 6299387 TI - 'Gelatinase-like' activity from articular chondrocytes in monolayer culture. AB - In addition to releasing collagenase and proteoglycanase activity, rabbit articular chondrocytes in monolayer culture released into the culture medium, latent, neutral enzyme activity which when activated by p-aminophenylmercuric acetate degraded fluorescein-labeled polymeric rat tail tendon Type I collagen and the tropocollagen TCA and TCB fragments of human Type II collagen into smaller peptides at 37 degrees C. Enzyme activity was abolished if p aminophenylmercuric acetate-activated culture medium was preincubated with 1.10 phenanthroline, a metal chelator. Thus, articular chondrocytes in monolayer culture are capable of producing neutral proteinases which acting together can result in complete degradation of tendon and cartilage collagen to small peptides. PMID- 6299388 TI - Resting potential of the mouse neuroblastoma cells. I. The presence of K+ channels activated at high K+ concentration but closed at low K+ concentration including the physiological concentration. PMID- 6299389 TI - Resting potential of the mouse neuroblastoma cells. II. Significant contribution of the surface potential to the resting potential of the cells under physiological conditions. PMID- 6299390 TI - Sex difference in cellular calcium metabolism of rat hepatocytes and in alpha adrenergic activation of glycogen phosphorylase. PMID- 6299391 TI - Potentiation by steroids of the beta-adrenergic agent-induced stimulation of cyclic AMP in isolated mouse thymocytes. AB - There is an increasing amount of evidence suggesting that glucocorticoids may modulate the responsiveness of various cell types to beta-adrenergic agents. In some systems, it has been shown, in addition, that steroids potentiate the elevation of cAMP induced by catecholamines. Little is known however of the mechanism underlying steroid action. We have studied this 'permissive action' in isolated thymocytes which have specific receptor sites for both glucocorticoids and beta-adrenergic agents. The glucocorticoid compound dexamethasone did not alter intracellular cAMP level but markedly enhanced the stimulation produced by isoproterenol. This effect was instantaneous and was still measurable at 10(-7) M dexamethasone. A similar potentiating action was observed in the presence of corticosterone but also in the presence of sex steroids. Determination of beta receptors after cell preincubation in the presence of dexamethasone showed that rapid alterations in beta-receptors are not involved in this permissive action. Experiments done in the presence of the calcium chelator, ethyleneglycol bis(beta aminoethyl ether)-N,N'-tetraacetic acid, suggest that dexamethasone action could be related to a modification of calcium mobilization. PMID- 6299392 TI - Viable mouse thymocytes as a model system for studying the onset of hormone induced cellular refractoriness. AB - Mouse thymocytes are characterized as a model cellular system for studying the onset of hormone-induced cellular refractoriness (desensitization). This system has the following combination of useful features. (a) The cells can be isolated without the use of digestive enzymes, avoiding possible damage to surface receptors or to other exposed membranal constituents. (b) They can be kept viable for several hours, a period during which both stimulation and desensitization get well under way. (c) They can be stimulated by a variety of hormones which function via cAMP (beta-agonists, prostaglandin E1 and specific thymic humoral factors). (d) Their desensitization is receptor-specific. (e) They can be readily ruptured under mild conditions so as to allow a physiologically relevant biochemical analysis of hormonal stimulation and desensitization. (f) The hormonal response of these cells can be monitored simultaneously by the activation of adenylate cyclase, by the intracellular level of cAMP, and by the activation of cAMP-dependent protein kinase (which functions as a metabolic sensor for cAMP). In this cellular system, desensitization does not involve processes such as the efflux of cAMP, the activation of cAMP-phosphodiesterase or the synthesis of a protein mediator. On the other hand, desensitization can be accounted for by a hormone-triggered inactivation of the adenylate cyclase system. The immediate desensitization of thymocytes is reversible and occurs without apparent loss of functional receptors. Continuous presence of hormone is shown to be required not only for triggering the chain of events which leads to the readily reversible desensitization, but also for the process which transfers the cells to the subsequent, 'locked' desensitized state. PMID- 6299393 TI - [Ionic mechanism of the Woodwors staircase]. AB - Analysis of single-chamber model of electromechanical coupling in the myocardial cell has shown that Woodwors staircase can be imitated in two cases: 1) stationary input current Ca2+ strongly exceeds the potential-dependent uptake of Ca2+ into the cell through the sarcolemma; 2) the action potential (AP) is shortened abruptly with an increase of the myocardium stimulation frequency. The experiments performed on a fragment of the frog heart ventricle supported the conclusions of the model. Blocking of Ca-channels with nifedipine (10(-6) g/mol) at the background of isotonic substitution of 70% of NaCl resulted in the development of "negative staircase" with an increase of stimulation rhythm. An abrupt shortening of AP after rest at joint action of adrenaline (10(-6) g/ml) and blocker of Ca-channels D-600 (10(-6) g/ml) was accompanied by Woodwors staircase. PMID- 6299394 TI - [Sodium specificity of the effect of cyclic nucleotides on the conductance of cytoplasmic membranes of the photoreceptor]. AB - The effect of cyclic nucleotides (3',5'-cAMP and 3',5'-cGMP) on the membrane of the retina rod outer segment under intracellular dialysis conditions was shown to possess sodium specificity, its value in the medium with normal Na+ content being 4-20 times above that in sodium free solution. The results obtained permit a conclusion that the effect observed is due to the increased conductance of the cytoplasmic membrane of the outer segment and support the assumption concerning the mediator role of cyclic nucleotides in the photoreceptor act. PMID- 6299395 TI - [Mechanism of guanosine-5'-monophosphate photodecomposition as affected by vacuum and laser ultraviolet irradiation]. PMID- 6299396 TI - [Low molecular channel-former protein from the mitochondrial membrane]. AB - Individual low molecular weight protein component was isolated by gel-filtration method from the inner mitochondrial membrane. This membrane component increases the conductivity of bilayer lipid membranes (BLM) in the presence of K+ and Ca2+ ions. This phenomenon may be explained by the formation of single conductivity channels. The voltage - current characteristics of this channel is nonlinear, which may be the result of asymmetrical operation of the channel - former in the polarized membrane, in spite of equal concentration of protein on both sides of the membrane. PMID- 6299397 TI - [Mechanism of the changes of EPR signals in leaves of higher plants during seedling development]. PMID- 6299398 TI - [Reversibility of the proton-potassium pump and synthesis of ATP in E. coli]. AB - E. coli is able to substitute 2H+ from the external medium for one K+ of a cell with the concurrent synthesis of ATP under certain condition. The 2H+/K+ exchange and coupled ATP synthesis are in need of both the essential H+ (around 24 KJ) and the high K+ gradient (10(3) ) directed from a cell to the medium. The reverse pump, cycle is acutuated as well as the direct one merely through an increase of osmolarity in the environmental medium. The reverse process can be blocked by an insignificant increase in external pH or in K+ concentration, by DCCD or by protonophore. The stoichiometry of the entire pump cycle is ATP:2H+:K+. PMID- 6299399 TI - [Study of human blood, erythrocytes and plasma with the EPR method at 77 degrees K]. PMID- 6299400 TI - [Electron-conformation transitions in the complex of bacterial photosynthetic reaction centers with cytochromes]. AB - A theoretical model of conformation--regulated electron transfer from multihaem cytochrome c to bacteriochlorophyll of the reaction centre (RC) is considered. The theoretical data are compared with the experimental ones on the basis of temperature dependence of laser-induced electron transfer from high-potential cytochrome Ch bacheriochlorophyll of RC in Ectothiohodospira shaposhnikovii chromatophores. From this comparison there were calculated the thermodynamic characteristics of cytochrome Ch transfer from the configuration without electron transfer to RC bacteriochlorophyll into the coordinated configuration with an effective transfer. The values obtained are: H = 7,1 kJ/M; S = --(30,2--36,9) J/grad. M. Possible regulatory role of such conformation transitions is discussed. PMID- 6299401 TI - Early increase of a 105,000-dalton phosphoprotein during meiotic maturation of Xenopus laevis oocyte. AB - In ovo [32P] phosphoproteins were analyzed during meiotic maturation of Xenopus laevis oocytes. A phosphoprotein of 105,000-dalton was found to increase early (one hour) after progesterone induction of meiosis. The pure heat-stable inhibitor (PKI) of cAMP-dependent protein kinase, which induces maturation, was microinjected into oocytes. Again the early increase in the 105,000-dalton [32P] phosphoprotein occurred. The burst in protein phosphorylation, which takes place at the period of germinal vesicle breakdown, was quantitatively and qualitatively comparable in progesterone and PKI-stimulated oocytes. In order to confirm the inverse relationship between the 105,000 dalton [32P] phosphoprotein increase and cAMP-dependent protein kinase activity, purified C-subunit of the kinase has been microinjected into oocytes. C-subunit which inhibits maturation did not increase significantly the 105,000-dalton [32P] phosphoprotein whereas it increased the total level of in ovo phosphorylation. Enucleation experiments favour the localization of the 105,000-dalton protein in both the oocyte cytoplasm and nucleus. Furthermore the progesterone-induced increase in the phosphorylation of the 105,000-dalton protein was found in the cytoplasmic compartment after oocyte enucleation. PMID- 6299402 TI - Evidence that a cAMP binding protein from Dictyostelium discoideum carries S adenosyl-L-homocysteine hydrolase activity. AB - A cAMP-adenosine binding protein partially purified from exponentially growing Dictyostelium discoideum cells carries S-adenosyl-L-homocysteine (SAH) hydrolase activity. This protein is present throughout the developmental cycle and has many properties in common with a cAMP binding activity previously reported from this laboratory (Gunzburg and Veron, 1981). Direct binding measurements with radioactive ligands indicate a dissociation constant of 0.2 microM for adenosine and 9 nM for cAMP, a value in good agreement with measurements of the rate constants for cAMP binding (k+1 = 2.4 X 10(4) M-1 sec-1) and dissociation (k-1 = 1.1 X 10(-4) sec-1). The binding of cAMP is completely abolished in the presence of 1 microM adenosine; a maximum 60 per cent inhibition of adenosine binding can be achieved with cAMP concentrations as high as 0.1 microM, suggesting that at least some of the cAMP and adenosine binding sites are not identical. The protein has a sedimentation coefficient of 9.2S and a native molecular weight of 190,000, as judged by gel filtration. Labeling with the photoaffinity ligand 8-azido-[3H] cAMP followed by SDS polyacrylamide gel electrophoresis results in a single band of 47,000 MW, suggesting that the protein may be a tetramer. The physiological importance of the protein and its association with SAH hydrolase activity is discussed in relation to a possible role in the regulation of protein and phospholipid methylation that occurs during chemotaxis. PMID- 6299403 TI - Cloning and expression of a DNA fragment carrying a his nifA fusion and the nifBQ operon from a nif constitutive mutant of Klebsiella pneumoniae. AB - From the nifc mutant plasmid pPC868, previously shown to carry a DNA duplication responsible for the Nifc phenotype, a 10 kb HindIII fragment was cloned into the multicopy vector pBR325. Restriction analysis of the resulting plasmids and in vitro deleted derivatives confirmed that the mutation was a fusion between his and nifLA. The order was hisG-hisD'-'nifL-nifA so that nifA was transcribed under the control of the his promoter and the nifL gene was altered. In addition the cloned fragment contained the adjacent nifBQ operon, and complementation data revealed that the nifA, nifB and hisG genes were expressed. Synthesis of nifA product under the transcription control of the his (or cat [CmR]) promoter enabled complementation of nifA and nifB mutations either in the absence or the presence of ammonia, but did not restore nitrogen fixation in a glnF mutant. Therefore, the nifA gene product requires glnF for its positive control function in a manner analogous to ntrC. Protein content analysis of minicells containing various multicopy nif plasmids confirmed the genetic organization mentioned above. A new polypeptide of 51,500 daltons was found whose synthesis was observed at 30 degrees C but not at 37 degrees C. According to the physical map, this protein could be the nifB gene product. Our results are in agreement with nifB transcription being under the control of a thermolabile nifA product. Moreover we obtained results suggesting that the presence of multiple copies of a functional nifB gene inhibited nitrogen fixation. PMID- 6299404 TI - Purification of human liver uridylyl transferase and comparison with the erythrocyte enzyme. AB - Uridylyl transferase (UDP glucose: alpha-D-galactose 1 phosphate uridylyl transferase, EC 2.7.7.12) has been purified 1350-fold from human liver to complete homogeneity. The purification procedure involved ammonium sulfate fractionation, batch treatment, chromatography on DEAE-cellulose, hexylagarose and hydroxylapatite. The specific activity of the homogeneous enzyme was 27 units/mg protein. The liver enzyme was compared to the red cell enzyme previously purified by us. The liver enzyme was similar to the red cell enzyme with respect to subunit molecular weight, kinetic studies and immunological properties. Differences in electrophoretic behaviour were found: the liver transferase has a more basic pI (between 5.5 and 5.8) than that of the erythrocyte enzyme (pI between 5.0 and 5.45). It is very likely that the liver uridylyl transferase and the red blood cell transferase are the same enzymes with post-translational modifications. PMID- 6299405 TI - A method for selecting the coding DNA for a protein of known sequence. II. Use of obligatory restriction sites. AB - A method for selecting the coding DNA of a protein of known sequence in a library of chimeric plasmids constructed with cDNA is described. It is based on the prediction of obligatory restriction sites within the searched cDNA. A partially purified probe can be obtained by taking advantage of the occurrence of two such obligatory sites. Another suggested possibility is to combine one obligatory restriction site and a specific synthetic oligodeoxynucleotide used as primer in classical reverse transcription methods. PMID- 6299406 TI - [Reverse electron transfer in mitochondria of the yeast Endomyces magnusii grown on sucrose]. AB - Some specific features of cytosol-mitochondria interactions during the growth of the Endomyces magnusii yeast on sucrose media were investigated. Under the given experimental conditions exogenous (cytoplasmic) NADH is the main source of reducing equivalents. Using low temperature EPR spectroscopy, it was demonstrated that exogenous NADH can effectively induce reverse electron transfer in the respiratory chain of the yeast. Fluorimetric assays suggest that End. magnusii mitochondria can utilize the reducing equivalents formed by reverse electron transfer for the reductive amination of alpha-ketoglutarate to form glutamate. The intramitochondrial localization of aminotransferases was postulated. PMID- 6299407 TI - [Redox-dependent protonation of cytochrome oxidase hemes in submitochondrial particles of the bovine heart]. AB - Potentiometric titrations of cytochrome oxidase in bovine heart submitochondrial particles were carried out within the pH range 5.3-9.0 in the alpha-band of heme absorption. The data obtained were analyzed within a model of non-interacting hemes and in accordance with a "neoclassical" scheme implying heme--heme interactions between cytochromes a and a3. The individual pH-dependencies of half reduction potentials of cytochromes a and a3 were determined. It was found that the redox-dependent protonation (the oxidation Bohr effect) is characteristic of the both cytochromes; however, the reduction of one of the hemes significantly diminishes the Bohr effect for the second heme. The redox transitions of cytochrome a are coupled to ionization of at least two heme-linked acid-base groups of the enzyme with pK1(red) in the acidic and pK2(ox) in the alkaline regions of pH, whereas the pH dependence of E0' for cytochrome a3 fits to the model containing one hemi-linked group with a pKred in the acidic region. PMID- 6299408 TI - Vanadium and affective disorders. PMID- 6299409 TI - Naloxone and self-mutilation. AB - A 15-year-old male with a long history of self-mutilation resembling the Lesch Nyhan syndrome, but with normal uric acid levels, was treated with naloxone. Pain inducing behavior decreased in the evenings following infusion of the drug, and was markedly reduced for 2 days following the treatment period. Though not recommended as therapy, the observations suggest that naloxone may play a role in the regulation of endorphin/enkephalin neural systems. PMID- 6299410 TI - Gonadotrophic stimulation of androgen secretion by the early fetal pig ovary in organ culture. AB - An influence of gonadotropins on steroid secretion by the early fetal ovary of the domestic pig was shown by organ culture and radioimmunoassay. Gonads from fetuses at Days 32-37 of gestation were cultivated singly for 9-12 days in biologically supplemented medium. One member of each pair of gonads was exposed to human chorionic gonadotropin (hCG) or ovine luteinizing hormone (LH), and the other served as a control. A marked stimulating effect on androgen secretion was noted with both gonadotropins. The major androgen found was androstenedione, with secretion rates of greater than 200 ng/gonad per 24 h for some explants exposed to hCG. Little or no androstenedione production occurred unless gonadotropin had been added to the culture medium. Lesser amounts of testosterone (usually less than 5% of the total of androstenedione and testosterone) were present. The data demonstrate a remarkable latent capacity for androgen biosynthesis by the early fetal pig ovary. PMID- 6299411 TI - A comparative study of the mechanism of action of luteinizing hormone and a gonadotropin releasing hormone analog on the ovary. AB - The mechanism of action of a gonadotropin releasing hormone (GnRH) agonistic analog ([D-Ala6]GnRH) on the rat ovary has been studied in comparison to similar effects of luteinizing hormone (LH). Stimulation of meiosis resumption in vitro in follicle-enclosed oocytes by both LH and [D-Ala6] GnRH, was blocked by elevated levels of cAMP as demonstrated when either dibutyryl cAMP or the phosphodiesterase inhibitor methylisobutylxanthine was present in the culture medium. In vivo, the prostaglandin synthase inhibitor indomethacin, which blocks LH-induced ovulation, also inhibited ovulation induced by the GnRH analog in hypophysectomized rats. On the other hand, the potent GnRH-antagonist [D-pGlu1, pClPhe2, D-Trp3,6] GnRH which blocked the stimulatory effect of the agonist on oocyte maturation and ovulation had no effect on LH action. It is concluded that while a GnRH-like peptide does not seem to mediate LH action on the ovarian follicles, both LH and GnRH agonist share some common mechanistic pathways at a post-receptor locus. PMID- 6299412 TI - Effects of hyper- and hypoprolactinemia on gonadotropin secretion, rat testicular luteinizing hormone/human chorionic gonadotropin receptors and testosterone production by isolated Leydig cells. AB - The effect of prolactin (Prl) on gonadotropin secretion, testicular luteinizing hormone (LH)/human chorionic gonadotropin (hCG) receptors, and testosterone (T) production by isolated Leydig cells has been studied in 60-day-old rats treated for 4 days, 4 and 8 weeks with sulpiride (SLP), a dopaminergic antagonist, or for 4 days and 4 weeks with bromocriptine (CB), a dopaminergic agonist. Plasma Prl concentrations were significantly greater in the SLP groups (204 +/- 6 ng/ml) and lower in the CB groups (3.0 +/- 0.2 ng/ml) than those measured in the control groups (54 +/- 6 ng/ml). The plasma concentrations of gonadotropin were not affected by a 4-day treatment with SLP or CB, nor were they after a 4-week treatment with CB. However, the hyperprolactinemia induced by an 8-week treatment with SLP was associated with a reduced secretion of gonadotropin (LH, 16 +/- 4 vs. 35 +/- 6 ng/ml; FSH, 166 +/- 12 vs. 307 +/- 14 ng/ml). In SLP-induced hyperprolactinemia, a 30% increase in the density of the LH/hCG testicular binding sites was observed (178 +/- 12 fmol/mg protein), whereas a 60% decrease was measured in hypoprolactinemia (55 +/- 5 vs. control 133 +/- 5 fmol/mg protein). Plasma T levels were increased in 4-day and 4-week hyperprolactinemic animals (4.3 +/- 0.4 and 3.9 +/- 0.4 ng/ml, respectively), but returned to normal levels in the 8-week group (3.0 +/- 0.5 vs. C: 2.3 +/- 0.2 ng/ml). No T modifications were observed in hypoprolactinemic animals. Two distinct populations of Leydig cells (I and II) were obtained by centrifugation of dispersed testicular cells on a 0-45% continuous Metrizamide gradient. Both possess LH/hCG binding sites. However, the T production from Leydig cells of population II increased in the presence of hCG, whereas that of cell population I which also contain immature germinal cells did not respond. The basal and stimulated T secretions from cell populations I and II obtained from CB-treated animals were similar to controls, whereas from 4 days to 8 weeks of hyperprolactinemia, basal and hCG induced T productions from cell population II decreased progressively. These data show that hyperprolactinemia causes, in a time-dependent manner, a trophic effect on the density of LH/hCG testicular receptors; reduces basal and hCG-stimulated T production from isolated Leydig cells type II; and results in an elevated plasma T concentration which decreases with time. The latter suggests a slower T catabolism and/or an impaired peripheral conversion of T into 5 alpha-dihydrotestosterone (DHT). Although hypoprolactinemia is associated with a marked reduction in testicular LH receptors, it does not affect T production. PMID- 6299413 TI - Potassium movement during sodium-induced motility initiation in the rat caudal epididymal spermatozoa. AB - The sodium-induced sperm motility initiation of the rat cauda epididymal sperm has been studied in different potassium concentrations. High K+ inhibited motility initiation. At a K+ concentration of 50 mM (concentration found in the rat cauda epididymidis), sperm motility was inhibited by 80%. K+ movement across the sperm membrane has been followed by using 86Rb+ as a marker for K+. When the 86Rb+ preloaded sperm were suspended in a sodium-free medium, there was a little efflux of 86Rb+. However, if they were suspended in a sodium-containing medium, the efflux rate was greatly increased. This increase in 86Rb+ efflux rate was associated with an initiation of sperm motility. Both 86Rb+ efflux and motility initiation were triggered by a K+ ionophore 18-crown-6 (2 X 10(-3)M). However, the ionophore-induced 86Rb+ efflux and motility initiation only occurred in the presence of extracellular Na+. Tetraethylammonium (TEA) chloride, which blocks K+ channels, inhibited motility initiation in a dose-dependent manner. Changes in the membrane potential of sperm have been followed using the fluorescent dye diO C6-(3) whose fluorescence in sperm suspension changes markedly with changes in sperm membrane potential. During motility initiation, there was a fall in fluorescence of the dye due to increased partition into sperm cells. This observation may indicate a hyperpolarization of the sperm membrane during motility initiation. It was concluded that sperm motility initiation is associated with a complex ionic event. Na+ enters sperm cells in exchange with H+ and K+. This change in the permeability of the sperm membrane to ions is reflected by a change in the sperm membrane potential. PMID- 6299414 TI - Studies on the mechanisms of hormone actions with macromolecular photoaffinity labeled receptors. PMID- 6299415 TI - Use of nucleotide photoaffinity probes to study hormone action. AB - It has been clearly shown that the action of several hormones is differentially mediated intracellularly by nucleotides containing either adenosine or guanosine base units. To study the protein-nucleotide interactions involved in several complex biological systems our laboratory has synthesized several 8-azido adenosine (8-N3 A) and 8-azidoguanosine (8-N3 G) derivatives of naturally occurring nucleotides. Modification of the nucleotides in the 8-position of the purine ring was done because: a) 8-substituted derivatives of cAMP and cGMP activated their respective protein kinases at physiological concentrations and were much less susceptible to hydrolysis by specific phosphodiesterases (PDE's) and b) substitution at the 8-position was much less likely to disturb the preferential and selective binding of adenosine versus guanosine nucleotides by enzymes that are specifically regulated by such interactions. This would allow studies of guanosine nucleotide specific binding in the presence of both adenosine nucleotides and adenosine nucleotide binding proteins, and vice-versa. In general, such has been the case and [32P] 8-N3 cAMP and [32P] 8-N3 cGMP have been used effectively to study their respectively activated protein kinases in several systems. Also, [32P] 8-N3 ATP has been used to study several ATPases and kinases while [gamma 32P] 8-N3 GTP has been shown effective for studies on tubulin and the G-regulatory protein (G/N) of adenylyl cyclase (A.C.). Several observations suggest that there must be important physical and energetic tie-ins between external hormone binding and the loading and unloading of specific internal nucleotide binding sites. These binding sites may be activator signals for protein kinases (e.g., cAMP protein kinase regulatory subunit), or cyclases (e.g., G/N proteins of A.C.) or catalytic sites involved in the production or hydrolysis of cyclic nucleotides. The thrust of this article is to detail the use of 8-azidopurine photoaffinity analogs of ATP, GTP, cAMP and cGMP as they may be used to study hormone-mediated events which may or may not involve cyclic nucleotides as a second messenger. PMID- 6299416 TI - Cyclic adenosine 3',5' monophosphate, calcium and protein phosphorylation in flagellar motility. AB - cAMP and calcium are two important regulators of sperm flagellar motility. cAMP stimulates sperm motility by activating cAMP-dependent protein kinase and catalyzing the phosphorylation of sperm proteins. The stimulation of sperm motility by cAMP appears to be at two different levels. Evidence has been presented to suggest that cAMP-dependent phosphorylations may be required in order for motility to be initiated. In addition, cAMP-dependent phosphorylation appears to modulate specific parameters of motility resulting in higher beat frequency or greater wave amplitude. Calcium, on the other hand, when elevated intracellularly to 10(-6) M or higher, inhibits flagellar motility. The calcium binding protein, calmodulin, appears to mediate a large number of effects of calcium on motility. Evidence suggests that calcium-calmodulin may be involved at the level of the membrane to pump calcium out of the flagellum. In addition, calcium-calmodulin may be involved in the control of axonemal function by regulating dynein ATPase and myosin light chain kinase activities. The identification of cAMP-dependent protein kinase, calmodulin and myosin light chain kinase in the sperm head suggests that cAMP and calcium-dependent phosphorylations are also involved in the control of the fertilization process, i.e., the acrosome reaction, in a manner similar to that known for the control of stimulus/secretion coupling. Finally, the effects of cAMP on flagellar motility are mediated by protein phosphorylation while the effects of calcium on motility are also in part, mediated by effects on protein phosphorylation. PMID- 6299418 TI - Simian virus 40-coded antigens and the detection of a 55K-dalton cellular protein in early embryo cells. PMID- 6299417 TI - Cytochrome c oxidase: two models. PMID- 6299419 TI - Molecular characteristics of brain opiate and nicotine receptors. AB - The present review has dealt with some of the molecular characteristics of the receptors associated with the action of opiates and nicotine. The binding characteristics of the opiate receptor in brain membrane preparations has been compared with those of a Triton X-100-solubilized extract of membranes as well as a preparation purified from rat brain by affinity chromatography. The purified opiate receptor appears to consist of three proteins; and the major component, which has a molecular weight of 35,000, may be the site of attachment of the opiate. Although phosphatidylserine appears to be associated with the opiate receptor within the membrane, its exact role in receptor binding is not known. The brain receptor for nicotine, which has a Kd in the nanomolar range, appears to have binding characteristics which differ from that of the classical nicotinic cholinergic receptor. This conclusion is corroborated by a characteristic behavioral response elicited by nicotine and not by cholinergic agonists. Finally, a discussion is presented on the presence of opiate receptors in human erythrocytes and leukocytes and the changes occurring in opiate addicts. PMID- 6299421 TI - Binding of heparan sulphate and heparin to control and virus-transformed cells. AB - A cloned embryonic mouse cell line contained specific cell-surface receptors for heparin and both the number and affinity appeared to be unchanged in a simian virus-40-transformed subclone. In competitive binding assays heparan sulphate from the control clone was bound preferentially compared to that from the transformed subclone, indicating that the altered sulphation of heparan sulphate from transformed cells results in a lowered affinity for cell-surface receptors. Evidence was obtained suggesting that endogenous proteoglycans were not held at the cell surface by binding to these receptors alone. However the possibility that proteoglycans embedded in the plasma membrane may interact with the receptor has not been ruled out. PMID- 6299420 TI - Effect of pH and different substrates on the electrokinetic properties of (Na+, K+)-ATPase vesicles. AB - Some biophysical properties of a (Na+, K+)-ATPase preparation from guinea-pig kidney have been analysed. The recently developed technique of laser Doppler spectroscopy was applied to measure particle mobility under electrophoretic conditions. The following results were obtained: 1. magnesium ions at pH 7.3 decrease the mobility of the ATPase containing vesicles by binding to negatively charged surface groups. At pH 3.3 the competitive binding of protons causes a shift of the mobility vs. [Mg2+] curve to higher values of [Mg2+], 2. binding of ATP at pH 7.3 (Kd = 0.9 X 10(-4) M for (mM 1 NaCl, 0.2 KCl, 0.1 MgCl2, 0.1 Tris) was measured as an increase in particle mobility depending also on [Mg2+]. At pH 3.3 also unspecific ATP-binding occurred, 3. ITP and GTP had the same Kd value as ATP; ADP a slightly lower one (Kd = 1.2 X 10(-4) M). Tris-H3PO4 (Kd = 2.6 X 10( 4) M) was also able to increase particle mobility, but only at higher concentrations and not to the same extent as ATP; AMP induced only very small changes, 4. from the mobility-pH curve an isoelectric point of 4.1 is derived (buffer: 1 mM NaCl, 0.2 mM KCl, 0.1 mM MgCl2, 0.1 mM Tris). In the presence of 0.9 mM ATP the isoelectric point is shifted to 3.2. As the electrophoretic mobility is directly proportional to the net charge of the vesicles, the results may be interpreted as changes in surface charge density, originating from both a conformational change of the ATPase polypeptide and a decrease in vesicle size. PMID- 6299422 TI - [Computer analysis of chromatin fragmentation with nucleases]. AB - Computerizing of the densitograms of chromatin hydrolysate DNA is offered. It is based on searching for gaussoids depicting the peaks of individual oligomers so that the resultant curve corresponded to the experimental curve to a high enough accuracy. The method suggested was used to study the time course of mononucleosome and oligomer accumulation during chromatin fragmentation with endogenous nuclease. Comparison of the experimental data to a model calculated for break down of 500-nucleosome chromatin fragments demonstrates that in chromatin, the release of mono-, di- and trinucleosomes is shifted towards nucleosomes during break down induced by endogenous nuclease. PMID- 6299423 TI - [Responses of the inspiratory and expiratory sites of giant cell nuclei to stimulation of more than two neurons]. PMID- 6299424 TI - [Types of cAMP-dependent protein kinase in the splenocytes of immunized animals]. PMID- 6299425 TI - [Contractility restitution during isotonic and isometric modes of myocardial contraction]. PMID- 6299426 TI - [Enzymes of cyclic nucleotide degradation in eye tissues during experimental ocular herpes]. AB - Activity of phosphodiesterases disintegrating cAMP and cGMP in the cornea, sclera and ciliary body was investigated in health and in different stages of experimental herpetic keratitis. The problems concerning the role of the cyclase system in the pathogenesis of herpetic keratitis and the possibility of applying some of the drugs to the disease treatment are discussed. PMID- 6299427 TI - [Molecular mechanism of combined effect of amphotericin B and cyclophosphamide on a virus-induced neoplastic process]. AB - Study of an experimental neoplastic process induced by virus type strains of Rous's sarcoma has shown that amphotericin B potentiates antineoplastic action of cyclophosphamide. Administration of cytostatin does not affect the fractional composition of chromatin proteins. Administration of polyene changes this indicator, the changes being greater after combined use of both drugs. The character of a considerable part of the abnormalities in the fractional composition of chromatin proteins indicates that chromatin had been exposed to proteolytic enzymes. PMID- 6299428 TI - [Ionic mechanism of the stimulatory effect of noradrenaline on smooth muscle cells of the pulmonary artery]. AB - Noradrenaline (NA) in a concentration of 5 X 10(-6) M produces depolarization of smooth muscle cells of the rabbit pulmonary artery and reduction of membrane resistance followed by contraction and increased excitability of muscle cells. Experiments with repolarization of the membrane exposed to NA in normal and Ca free Krebs solutions have shown that activation of the NA-induced contraction is mainly due to Ca++ entering the cells through NA-sensitive potential-dependent Ca channels. The NA-induced depolarization results from an initial decrease of K permeability of the membrane subsequent increase of the permeability of NA sensitive potential-dependent channels for Na+ and/or Cl-, which provides further depolarization of the membrane. Depolarization ceases after becoming sufficient for activation of potential-dependent non-inactivated K-channels. Voltage clamp experiments have shown that the NA-induced increased excitability is related to a reduction of slow, particularly of fast component of outward current, whose early activation prevents the development of regenerative process of action potential generation under normal conditions. PMID- 6299429 TI - [Genetic control of the ACTH content of mouse plasma]. AB - The content of ACTH in blood plasma of C57BL/6, BALB/C, F1, F2, and F1 X BALB/C mice was measured before and after an open-field exposure to emotional stress. Differences in the initial content of hormone and its changes after stress were revealed as dependent on the animal line. A suggestion is made about monogenic control over the initial level of ACTH. PMID- 6299430 TI - [Stress analgesia in experimental burn shock]. AB - Experiments with curarized and slightly anesthetised adult cats with burn shock have demonstrated pronounced depression of excitation transmission in associative and nonspecific afferent systems during somatic, sound and light stimulation. Effects controlling the activity of the cardiovascular system were facilitated in efferent systems. In white rats, burn shock led to an increase in the somatic and visceral pain threshold during the first 5 days. Rausedyl and naloxone reduced stress analgesia caused by burn shock in white rats. Burn shock enhanced opiate like activity (beta-endorphine, beta-lipotropin) of the white rat forebrain as shown by radioimmunoassay. The data suggest that stress analgesia associated with experimental burn shock is likely to be accounted for by the increased production of endogenous opiates. PMID- 6299431 TI - [Dynamic study of the development of edema in the organs in experimental burn trauma]. AB - Proton magnetic relaxation with measuring the time of spin-screen relaxation (T1) was used to study the time course of derangement of organ and tissue flooding after stage IIIB experimental burn of 15% body area. To examine the structures of the microcirculatory vessels, use was made of electron microscopy of the organs. The internal organs under study demonstrated a significant long-term (up to 49 days) increase in flooding: in the liver and kidneys, it occurred since the first day after burn, while in the lungs, myocardium and brain, since the 7th day. The results indicate the parallelism of the changes in the T1 and electron microscopic alterations in the histohematic barriers. Proton magnetic relaxation enables a more rapid and more convenient assay of the changes in tissue flooding as compared to the method of drying at high temperature. Therefore, it may be applied to the assessment of the efficacy of different therapeutic means. PMID- 6299432 TI - [Convulsions induced by kynurenine and quinolinic acid as a sensitive test for assessing the anticonvulsant activity of GABA-ergic preparations]. AB - Phenibut, sodium hydroxybutyrate and baclofen are selectively effective against seizures induced in mice by the endogenous metabolites of tryptophan, L kynurenine and quinolinic acid. The seizures were not affected by the drugs in doses under study. Depakine and aminooxyacetic acid as well as diazepam and phenobarbital appeared the most effective against pentylenetetrazole seizures. GABA and muscimol administered intracerebroventricularly merely prolonged the latency of seizures. Dissimilarities in the GABA-ergic mechanisms of the anticonvulsant effects of the drugs under consideration are discussed. PMID- 6299433 TI - [Stimulus-dependent blockade of the Na channels of isolated myocardial cells in the rat by the anti-arrhythmia agent N-propyl ajmaline (neogiluritmal)]. AB - The patch-clamp method was used to study the effect of N-propyl ajmaline (NPA) on sodium currents (INa) in enzymatically isolated cells of adult rats. When applied at concentrations of 10(-5)--2 X 10(-5) M NPA produced a use-dependent inhibition of INa. The rate and depth of this blockade increased with NPA concentration, frequency and magnitude of depolarizing pulses. Voltage dependence of the blockade was revealed beyond the potential range where the fraction of activated Na channels was close to 1.0. Turning on the strong hyperpolarizing prepulse (100 ms duration) before each test pulse induced a prompt increase of INa (due to the removal of resting inactivation) which declined pulse by pulse during subsequent repetitive stimulation. A single long-lasting (3000 ms) depolarizing step, in contrast to the train of short (5 ms) depolarizing pulses did not enhance appreciably the blocking action of NPA. From an analysis of the data obtained it was inferred that NPA inhibits INa due to a voltage-dependent interaction with open Na channels. It is emphasized that NPA has similar blocking effects on Na channels in myocardial cells and nerve fibers. PMID- 6299434 TI - [Range of the transmissivity of the genetic transfer factors pAP38, pAP39, pAP41 and pAP42]. AB - A study was made of the transmissivity range of the genetic transfer factors pAP38, pAP39, pAP41 and pAP42 identified in E. coli. It was demonstrated that these factors are not capable of transfer to the cells of P. putida, P. fluorescens, R. leguminosarum, A. lipoferum, A. tumefaciens. Factor pAP42 is mobilized to transfer to P. putida and R. leguminosarum with the aid of plasmid RP4. It is assumed that in the course of mobilization, the cointegrative structures are formed between plasmids pAP42 and RP4. PMID- 6299435 TI - Macromolecules stimulating human granulocytic colony-forming cells, precursors of these cells, and primitive erythroid progenitors: some apparent nonidentities. AB - The relationship between molecules having granulocyte colony-stimulating activity (G-CSA), erythroid burst-promoting activity (E-BPA), and activity promoting increase in the number of granulocytic progenitors in liquid culture (delta GPA) was explored in conditioned medium from human leukocytes (HLCM) and human placenta (HPCM). As tested on human hemopoietic progenitors in culture, G-CSA eluted from Sephadex G100 as a single peak with apparent molecular weight of 25,000, separating partially from E-BPA and delta GPA, which both had an apparent molecular weight of 45,000. All three activities eluted together from hydroxyapatite at low molarity phosphate. Their charge properties were also similar and all three electrofocused in flat gel beds in the pH range near 5.4. On both hydroxyapatite and isoelectric focusing, delta GPA sometimes separated partially from the other two activities but not consistently. The gel filtration result shows that in conditioned medium of human origin, molecules having G-CSA are not the same as those having delta GPA, suggesting a dual factor requirement in the granulocytic lineage reminiscent of that in the erythroid pathway. The results suggesting that delta GPA might differ from E-BPA, on the other hand, were not consistent enough to establish their nonidentity. Single micromanipulated cells proved capable of forming erythroid or granulocytic colonies in the presence of either crude or partially purified activity. The results establish that human colony-forming cells are direct primary targets of growth factors in HLCM and HPCM. PMID- 6299436 TI - Characteristics of plasmalemma alkaline phosphatase of rat mesenteric artery. AB - General characteristics of alkaline phosphatase activity of the plasma membrane enriched fraction isolated from rat mesenteric arteries were investigated. The vascular smooth muscle plasmalemma alkaline phosphatase is a metalloenzyme which is strongly inhibited by chelating agents and this inhibition can be completely overcome by addition of Mg2+ or Ca2+. Zn2+ only partially reactivates the enzyme in the presence of low concentrations of EDTA. The enzymatic hydrolysis of p nitrophenyl phosphate, beta-glycerophosphate, alpha-glycerophosphate, or 3' adenosine monophosphate showed an optimal activity in the alkaline region between pH 9 and 11. The alkaline phosphatase activity is distinctly different from the plasmalemma ATPase and 5'-nucleotidase activities with respect to their pH dependence, influence by added divalent metal ions and stability against heat inactivation. Vanadate ion, being structurally similar to the transition state analog of the phosphoryl group, potently inhibits alkaline phosphatase with an apparent Ki of 1.5 microM. The altered alkaline phosphatase activity of vascular smooth muscle in relation to its possible physiological function and pathophysiological manifestation associated with hypertensive disease are discussed. PMID- 6299437 TI - Independent expression of the surface markers 5'-nucleotidase and cALLA on leukemic cells. AB - High levels of the ectozyme 5'-nucleotidase (5'-N) and the common ALL-antigen (cALLA) are coexpressed on leukemic blast cells in common ALL, in the lymphoid blast crisis of CML and also on the lymphoblastoid cell-line Nalm-1. Clinically this coexpression can help to subclassify leukemias and may be of diagnostic and prognostic significance. In an attempt to study the mechanism underlying this simultaneous expression plasmamembrane subfractionation was undertaken on Nalm-1. When membrane-shedding from intact cells is induced by sublytic concentrations of the lysophosphatidyl-choline analogue ET-12-H, membrane subfractions are obtained which contain 30-40% of total cellular 5'-N, which is most of the enzyme carried on the cell surface, in a highly enriched form. Under these conditions only a very low release of intracellular enzymes is observed. On the other hand cALLA is not accumulated in these membrane fractions to any appreciable extent. The predominant part of this antigen is still on the intact cells remaining after the shedding procedure. It is concluded that the simultaneous expression of 5'-N and cALLA on Nalm-1 and leukemic blasts is not regulated by a physical association or a close neighborhood of these antigens on the membrane level. PMID- 6299438 TI - [Dietary behavior, way of life, and nasopharyngeal cancer]. AB - Nasopharyngeal carcinoma is prevalent in Southeast Asia, Maghreb countries and Greenland/Alaska. This cancer, wherever prevalent, is closely associated to the Epstein-Barr virus (EBV), but its geographical repartition also suggests the possibility of a cultural factor. A comparison of food habits and way of life of Cantonese, Maghreb populations and Eskimos shows certain common points: dried fish or meat, fermented and sometimes rotten, as well as salt preserved vegetables. Certain berries of Euphorbiaceae are eaten by children and the plant itself is used in traditional medicine. These Euphorbiacea contain cancer promoting agents, able to reactivate the EBV latency. The hypothesis proposed here is that cancerous effects common to the different regions at high risk for nasopharyngeal carcinoma exists and that they either act separately or reactivate the EBV latency. PMID- 6299439 TI - PCB, DDT, and benzo(a)pyrene in raw and pan-fried white croaker (Genyonemus lineatus). PMID- 6299440 TI - [2 esteroproteases from mouse submandibular gland]. PMID- 6299441 TI - [Neuromuscular transmission considered from a structural and cytochemical viewpoint]. PMID- 6299442 TI - [Theory of neurohumoral transmission of nerve impulses. Its importance in the treatment of cardiovascular diseases]. PMID- 6299443 TI - Binding of yohimbine stereoisomers to alpha-adrenoceptors in rat liver and human platelets. AB - 1--Displacement of tritiated prazosin binding to rat liver plasma membranes and tritiated yohimbine human platelet membranes shows that (+)-yohimbine, alloyohimbine and alpha-yohimbine (rauwolscine) are selective alpha 2 adrenoceptor antagonists (KD alpha 1/KD alpha 2:635, 46.6 and 112 respectively) whereas corynanthine is more alpha 1-selective (KD alpha 1/KD alpha 2:0.036). 2- 11-Methoxy derivatives of alpha-yohimbine and epi-alpha-yohimbine are very weak alpha-adrenoceptor blockers. 3--It is concluded that the aromatic A ring, the Nb atom, and the carboxymethyl moiety are important for the binding of yohimbine to the alpha-adrenoceptor, the carboxymethyl group being important for the alpha 1/alpha 2 specificity of the molecule. PMID- 6299444 TI - An electrophysiological study of presynaptic alpha-adrenoceptors in the vas deferens of the mouse. AB - 1--Effects of clonidine and alpha-adrenoceptor antagonists were studied on sympathetic neuroeffector transmission in the mouse vas deferens. The amplitude of excitatory junction potentials (e.j.ps) was taken as a measure of transmitter release per impulse. 2--At a concentration of 0.5 microM, prazosin abolished depolarizations evoked by iontophoretically applied noradrenaline, but changed neither spontaneous nor nerve stimulation-evoked e.j.ps. 3--Yohimbine 0.1 and 1 microM, rauwolscine 1 microM and corynanthine 1 microM di not change the e.j.p. amplitudes elicited by the first 2-3 pulses in trains of 15 pulses at 3 Hz, but increased the e.j.ps elicited by the subsequent pulses. Corynanthine 1 microM was much less effective than yohimbine 1 microM or rauwolscine 1 microM, and corynanthine 0.1 microM had no effect. 4--Clonidine 0.01 microM reduced the e.j.p. amplitudes evoked by single pulses and its effect was counteracted by yohimbine 1 microM. 5--In vasa deferentia from reserpine-treated mice the e.j.p. trains were changed in much the same way as by yohimbine and rauwolscine. Yohimbine 1 microM did not further increase the e.j.p. amplitudes in these organs, whereas clonidine 0.01 microM caused a marked inhibition. 6--It is concluded that the release of the motor transmitter in the mouse vas deferens is inhibited by activation of presynaptic alpha-adrenoceptors, and that these receptors are normally activated by neurally released noradrenaline. PMID- 6299445 TI - Evidence that ionic channels associated with the muscarinic receptor of smooth muscle may admit calcium. AB - 1 The actions of carbachol on the membrane potential and conductance of smooth muscle of the guinea-pig intestine were investigated using microelectrode recording and the double sucrose-gap method in solutions in which calcium was the only cation creating an inwardly-directed electrochemical gradient.2 In a calcium chloride solution containing a small amount of potassium but no sodium and buffered to physiological pH (Ca Locke) the membrane was hyperpolarized to more than -80 mV. Carbachol (2 x 10(-7)-10(-4)M) depolarized the membrane and increased the membrane conductance.3 By passing current the membrane potential of the smooth muscle cells could be varied. In Ca Locke the depolarization produced by carbachol was shown to be reduced if the membrane was depolarized. The relationship between the size of the carbachol depolarization and the membrane depolarization was linear, giving an apparent reversal potential for carbachol depolarization some 20 mV positive to the resting membrane potential, as measured by extracellular electrodes in the sucrose gap.4 Carbachol depolarization was reduced if the calcium concentration was reduced below 2.5 mM by replacing calcium with Tris, but the depolarization in 2.5 mM Ca and in Ca Locke (100 mM Ca) were of similar size. In Ca-free Na-free solution with EGTA, carbachol depolarization was soon abolished.5 In the sucrose-gap when the chloride gradient across the cell membrane was reversed by replacing the chloride of Ca Locke by an impermeant anion the membrane depolarized. Carbachol now contracted the muscle but produced a hyperpolarization.6 These results are consistent with the hypothesis that activation of muscarinic receptors opens ionic channels which at least in the solutions used, can admit sufficient calcium ions to depolarize the cell and cause tension development. PMID- 6299447 TI - Persisting nutritional neuropathy in former war prisoners. PMID- 6299446 TI - Effect of steroids on beta-adrenoceptor-mediated relaxation of pig bronchus. AB - 1--Progesterone, testosterone (40 microM), cortisol and cortisol hemisuccinate (80 microM) caused 6-8 fold potentiations of (+/-)-isoprenaline (Iso)-induced relaxations of pig bronchus while several other steroids caused smaller potentiations or had no effect. 2--17 beta-Oestradiol (40 microM) increased the potency of Iso, (-)-adrenaline (Adr) and (-)-noradrenaline (NA) by 10.6, 2.3 and 2.6 fold respectively but had no significant effect on the potency of fenoterol (Fen). 3--Inhibition of catechol-O-methyl transferase (COMT) with U-0521 (30 microM) caused a 6 fold increase in the potency of Iso but failed to alter the potency of Adr, NA or Fen. The extraneuronal uptake inhibitor normetanephrine (50 microM) caused significant 2 fold increases in the potency of Iso and Adr but did not potentiate the responses to NA or Fen. 4--In preparations where the potency of Iso had already been increased by U-0521 (30 microM) or by normetanephrine, 17 beta-oestradiol produced no significant further increase in potency. These results indicate that steroid-induced increases in the potency of catecholamines in pig bronchus can be explained in terms of inhibition of COMT or extraneuronal uptake or both. PMID- 6299448 TI - H2 receptor antagonists-cimetidine and ranitidine. PMID- 6299449 TI - Single 1 g dose of cefotaxime in the treatment of infections due to penicillinase producing strains of Neisseria gonorrhoeae. AB - One hundred and two patients with an uncomplicated infection due to penicillinase producing strains of Neisseria gonorrhoeae (PPNG) were treated with a single 1 g dose of cefotaxime. At follow-up within 15 days all genital and rectal infections were cured. Pharyngeal infections also seemed to respond to this treatment. A relatively high proportion (30.9%) of patients, however, developed post gonococcal urethritis. PMID- 6299450 TI - Abnormalities of the uterine cervix in women with vulval warts. A preliminary communication. AB - Fifty women attending a clinic for sexually transmitted diseases with vulval condylomata acuminata were examined by cervical cytology and colposcopy for cervical infection by human papillomavirus (HPV) or epithelial abnormality indicating cervical intraepithelial neoplasia (CIN) or both. Collated results showed a high prevalence of both conditions in these 50 women; 25 (50%) had evidence of cervical infection by HPV and 18 (36%) epithelial abnormalities consistent with CIN 1 or 2. PMID- 6299452 TI - Antagonism of synaptic transmission in vivo: contributions of microiontophoresis. AB - Fundamental to the problems in the study of synaptic transmission in the mammalian central nervous system is the identification of the nature and role of transmitter substances at prescribed loci. Although recently many advances have been made in determining which compounds are endogenous to the central nervous system and which of these and other molecules alter neuronal activity, there can be little doubt that at the present level of our understanding of such brain constituent we are encountering merely a few of the basic materials requisite for the normal functioning of central neurones. Some 60 or 70 substances have now been postulated as candidates for involvement in synaptic processes. Many of the receptors for these and closely related compounds have been characterized pharmacologically through the examination of the effects of agonists and antagonists. One technique which has played a principal role in these neuropharmacological investigations is the method of local drug administration termed microiontophoresis. Although this method is invaluable for helping to answer questions at the molecular and membrane level, under circumscribed conditions its advantages may be exploited for use in experiments oriented toward a more physiological level of investigation. This short account is an attempt to provide a description of some of these conditions and further will describe some of the creative ways in which microiontophoresis has been utilized in vivo to reveal mechanisms by which the central nervous system processes information at the synaptic level. PMID- 6299451 TI - Immunocompromise syndrome in homosexual men. Prevalence of possible risk factors and screening for the prodrome using an accurate white cell count. AB - The immunocompromise syndrome in homosexual men in the USA is thought to be associated with cytomegalovirus (CMV) infection and nitrite intake. Such men often have a lymphopenia. In a clinic in London 76% of 46 unselected homosexual men and 50% of 76 heterosexual controls had serum CMV IgG antibody at a titre of 1/16 or more (p less than 0.01). No case of excretion of CMV in the urine was found. Thirty per cent of the homosexual men admitted to using nitrites. These figures suggest that this population of homosexual men uses nitrites less often than their counterparts in the USA and is less likely to have evidence of active or past CMV infection. In addition, these male homosexual patients seem to be less promiscuous than those reported from the USA. Mean accurate total and differential white blood cell counts, using the Haemalog D automatic white cell counter, were no different in homosexual men (and various at risk subgroups of them) than heterosexuals, suggesting that the prodrome to the immunocompromise syndrome was not present in the London clinic population. PMID- 6299453 TI - Short-term potentiation phenomena in the rat limbic forebrain. AB - Several types of short-term postactivation potentiation (PAP) effect were examined in limbic forebrain pathways in the chronic rat. We tested 9 different stimulation sites and 1-3 target sites for each stimulation site. All pathways showed PAP effects following activation by single electrical pulses or pulse trains. Using exponential curve fitting procedures, we found that the decay curves could be best fitted by one, or a sum of 2-3, exponential curves. On the basis of time constants, these curves fell into 4 different categories: facilitation (tau = 80 ms), augmentation (tau = 7s), potentiation 1 (tau = 70s), and potentiation 2(tau = 6.5 min). The latter component was the one most reliably generated in the chronic preparation. Frequency facilitation (facilitation during a stimulation train) was also examined and it appeared to be based upon a mechanism similar to that underlying paired pulse facilitation. Evidence is presented that facilitation and augmentation may be based on the the same mechanism. The possibility that the remaining components involve different mechanisms is discussed. PMID- 6299454 TI - Long-term potentiation phenomena in the rat limbic forebrain. AB - Long-term potentiation (LTP) phenomena were investigated in several limbic forebrain pathways. With the possible exception of the lateral olfactory tract, LTP could be produced in all pathways tested. LTP effects tended to increase as the stimulation site moved caudally along the pyriform lobe. The largest effects were produced by stimulation of pathways into and out of the hippocampus. Target sites also differed, with the hippocampal sites showing the strongest and longest lasting LTP effects. The time course of LTP appeared to be best fitted by the sum of two exponential curves with time constants of about 1.5 h and 5 days respectively. We also looked at the potentiation effects produced by repeated epileptogenic (kindling) stimulations, and the effect of this potentiation on subsequent tests of short-term and long-term potentiation. In most cases, the short-term effects, and the first component of the LTP effect, were still intact after kindling. The longest lasting component, however, could no longer be produced with either amygdala or perforant path stimulation. This result indicates that the potentiation produced by kindling may be based upon the same mechanism as the LTP effect. PMID- 6299456 TI - Effects of taurine and GABA on Ca spikes and Na spikes in cerebellar purkinje cells in vitro: intrasomatic study. AB - For the purpose of throwing some light on the inhibitory neuronal mechanisms in the cerebellum, particularly that by interneurons, the effects of taurine, a strong candidate for the transmitter of stellate interneurons, on spontaneous and evoked Ca spikes and Na spikes in Purkinje cells in guinea pig cerebellar slices were intrasomatically investigated in comparison with GABA, the transmitter of basket interneurons. In common, both amino acids hyperpolarized Purkinje cell membrane, inhibited somatic Na spikes and dendritic Ca spikes, and decreased somatic membrane resistance. Taurine applied focally onto Purkinje cell dendrites, however, was found to block Ca spikes and decrease the membrane resistance more potently than when it was applied onto the soma. GABA applied onto Purkinje cell somata was more potent in blocking somatic Na spikes and in decreasing the membrane resistance than taurine applied onto the somata. The dendritic sites of Purkinje cells, which were sensitive to iontophoretically applied taurine, responded also to focal electrical stimulation in an inhibitory manner. These results not only suggest that Purkinje cell dendrites are the primary site of action for taurine, but also support the previous concept that taurine acts as a transmitter in the dendritic interneuron, stellate cells. PMID- 6299455 TI - On the mechanism of action of GABA in pelvic vesical ganglia: biphasic responses evoked by two opposing actions on membrane conductance. AB - Intracellular recording techniques were used to study the response of cat vesical pelvic ganglion neurones loaded with permeable anions to the application of GABA in vitro. In 106/127 neurones GABA evoked a biphasic response, the initial phase of which was depolarizing and associated with a conductance increase; the latter phase was hyperpolarizing and associated with a conductance decrease. The GABA evoked hyperpolarization and conductance decrease were related and behaved as though generated by closure of ion channels open in the resting membrane. The hyperpolarization had a strong inhibitory action on both spontaneous activity, and excitation evoked by depolarizing current injection and pre-synaptic nerve stimulation. Ion substitution experiments suggest that the conductance decrease is primarily to chloride ions, although other ionic species may contribute. Short iontophoretic applications of GABA-evoked monophasic depolarizing excitatory responses, even during the hyperpolarizing response evoked by perfusion of GABA, suggesting no cross-desensitization between the mechanisms generating the initial and late phases of the biphasic response. PMID- 6299458 TI - Analgesia induced by electroconvulsive shock: brain enkephalins may mediate tolerance but not the induction of analgesia. AB - Electroconvulsive shock (ECS) administered to rats produces potent analgesia. This analgesic effect displays opiate characteristics with tolerance to ECS analgesia seen with repeated induction and cross tolerance observed following the chronic administration of analgesic dose of morphine. Furthermore, tolerance development after repeated ECS application could be blocked by daily pretreatment with the long acting opiate antagonist naltrexone (5 mg/kg). In contrast to these opiate characteristics of ECS analgesia, naltrexone administration (0.5, 5 and 10 mg/kg) failed to attenuate the acute analgesic effect of ECS. Measurement of brain enkephalin levels revealed an increase in enkephalin contents after 10 daily ECS applications, an effect which was not affected by naltrexone pretreatment. Acute administration of ECS did not affect brain enkephalin levels either immediately or 24 h after ECS administration. Significant increases in brain enkephalin following chronic ECS were observed in the hypothalamus and striatum but not in the mesencephalon or hindbrain. A gradual return of enkephalin levels to baseline values was seen with days when ECS was no longer administered. Parallel behavioral studies showed that ECS analgesia also showed a gradual recovery from tolerance after discontinuation of chronic ECS. Based on these and previous studies we propose that different systems may mediate the acute analgesic effect of ECS on the one hand, and the tolerance to such an effect observed after repeated administration on the other. We further suggest that the acute action of ECS may be mediated by substances with only partial opiate characteristics, while the long term effects observed may be due to the increase in brain enkephalin contents after chronic ECS application. PMID- 6299457 TI - Ionic mechanisms of the action of taurine on cerebellar Purkinje cell dendrites in vitro: intradendritic study. AB - The ionic mechanisms underlying the inhibitory action of taurine on Purkinje cell dendrites in guinea pig cerebellar slices were investigated with the Ca spikes intracellularly recorded from the dendrites of Purkinje cells. Taurine-induced hyperpolarization was found to be inverted at a membrane potential of about--125 mV and was largely reduced by the decrease of external Cl- concentration. As the hyperpolarization was associated with an increase in a membrane conductance, it was suggested that taurine hyperpolarizes the Purkinje cell dendrite primarily by increasing a Cl conductance. It was also found that taurine greatly flattened the rising phase of the Ca spike, suggesting that taurine may abolish Ca spikes by blocking the entry of Ca ions into Purkinje cell dendrites. The after hyperpolarization following each Ca spike was also found to be decreased by taurine, probably as a consequence of the blockade of the entry of Ca ions into the dendrite. This taurine-induced decrease of the after-hyperpolarization appears to be responsible for the transient small depolarization which usually occurs before Ca spikes are abolished by taurine. PMID- 6299459 TI - Differences in uptake kinetics of cis-3-aminocyclohexane carboxylic acid into neurons and astrocytes in primary cultures. AB - The uptake of [3H]ACHC and [3H]GABA into cultured neurons and astrocytes was studied. [3H]ACHC uptake was less efficient than that of GABA in both cell types and Km values for ACHC uptake into neurons and astrocytes were 40.3 microM and 210.8 microM, respectively. The corresponding Vmax values were 0.321 and 0.405 nmol . min-1 . mg-1 cell protein, respectively. Kinetic studies of the effects of GABA on ACHC uptake and vice versa showed that GABA is a linear competitive inhibitor of ACHC uptake in both cell types with a Ki value of 15 microM. On the other hand, ACHC turned out to be a complex inhibitor of astrocytic GABA uptake being competitive at lower concentrations and non-competitive at higher concentrations. ACHC inhibited GABA uptake into neurons competitively with a Ki of 69 microM. It is concluded that ACHC acts primarily on neuronal GABA uptake sites but its uptake is much more complicated than hitherto anticipated. PMID- 6299460 TI - Role of medial forebrain bundle catecholaminergic fibers in the modulation of glucocorticoid negative feedback effects. AB - The possible role of medial forebrain bundle (MFB) catecholaminergic fibers in the previously observed MFB-mediated modulation of hypothalamic sensitivity to glucocorticoid negative feedback effects has been investigated. MFB catecholaminergic fibers were destroyed by bilateral MFB injections of 6 hydroxydopamine (6-OHDA) to adult male rats. The inhibitory effect of dexamethasone (10 micrograms, i.p., 4 h) upon the adrenocortical secretory response to ether stress, as estimated by serum corticosterone levels, was enhanced in the 6-OHDA-treated animals, as compared to vehicle-injected controls. This study suggests that MFB noradrenergic neurons are at least partially responsible for the MFB-mediated modulation of hypothalamic sensitivity to glucocorticoid hormones. PMID- 6299461 TI - Enkephalins induce a centrally mediated rise in blood pressure in rats. AB - The cardiovascular effects of enkephalins after administration directly into the nucleus tractus solitarii (NTS) of urethane anesthetized rats were investigated. Unilateral microinjection of Met-enkephalin and its stable analogue D-Ala2-Met5 enkephalin resulted in a dose-related rise in mean arterial pressure which in the case of the analogue was accompanied by tachycardia. The elevation in blood pressure was anatomically specific and restricted to the intermediate third of the NTS, as verified histologically. These cardiovascular changes were prevented by pretreatment with locally applied naloxone (10 ng). A similar pressor effect was obtained with Leu-enkephalin. Antiserum to Met-enkephalin (1:50 dilution) caused a fall in blood pressure on injection into the NTS, and completely blocked the pressor response and tachycardia induced by D-Ala2-Met5-enkephalin. These results suggest that enkephalins have a pressor role in the central nervous system. PMID- 6299462 TI - Stimulation of brain Na+, K+-ATPase by norepinephrine in vivo: prevention by receptor antagonists and enhancement by repeated stimulation. AB - The role of alpha 1- and beta-noradrenergic receptors in the stimulation of the K+-p-nitrophenylphosphatase activity and ouabain binding associated with rat brain Na+, K+-ATPase by acute or repeated yohimbine were examined. Repeated yohimbine increased enzyme activity and ouabain binding by about 25%; this was prevented by the alpha 1-receptor antagonist prazosin and decreased by the beta antagonist propranolol. Pretreatment with repeated yohimbine increased the stimulation of enzyme activity by acute yohimbine while the stimulation was prevented by prazosin pretreatment. PMID- 6299463 TI - Quisqualic acid excitation of cortical neurones is selectively antagonized by streptomycin. AB - Although evidence exists for at least 3 kinds of excitatory amino acid receptor in the CNS, responding to N-methyl-D-aspartic acid, kainic acid and quisqualic acid, respectively, only antagonists at the former two sites are currently available. It is now reported that when applied by microiontophoresis to neurones in the rat cerebral cortex, excitatory responses to quisqualic acid can be selectively reduced by streptomycin. PMID- 6299464 TI - The effect of impulse blockage on cytochrome oxidase activity in the cat visual system. PMID- 6299465 TI - Circadian variations in beta-endorphin concentrations in pituitary and in some brain nuclei of the adult male rat. AB - Using both the 'punch' microdissection and a radioimmunological technique, circadian variations in beta-endorphin concentrations can be observed in the pituitary and in some discrete brain regions of the male rat (Wistar CFY). Animals were synchronized with light from 06.00 to 18.00 h, then darkness. Water and food were available ad libitum. Very well marked circadian rhythms were in evidence in the anterior lobe of the pituitary, the septum, the pons, the medulla oblongata and the cerebellum. There crest time locations were situated between 20.00 and 24.00 h. No significant circadian rhythms but biphasic variations were observed in the intermediate lobe of the pituitary, the POA, the thalamus, the central gray and the caudatus. There crest time locations were synchronized around 08.00 and 20.00 h. The most striking finding was that, regardless of the brain area investigated so far, maximal values were observed a short time after the beginning of the activity period of rats. This fact is identical with the one which has been observed for substance P and LH-RH contents in brain areas where these peptides are mostly present in nerve terminals in high concentrations. PMID- 6299466 TI - Characterization of beta-adrenergic receptors in rat brain and pituitary using a new high-affinity ligand, [125I]iodocyanopindolol. AB - The binding of [125I]iodocyanopindolol (ICYP) to membrane preparations from rat cerebral cortex, hypothalamus and anterior pituitary gland was characterized in regard to specificity, density, and the proportion of beta-adrenergic receptor subtypes. By employing a mixture of ligands specific for alpha-adrenergic, serotoninergic and dopaminergic receptors, it was possible to eliminate most of the less-specific contributions to ICYP binding profiles, which resulted in narrowing the range of measured dissociation constants to 35-50 pM for all neural tissues studied. These values corresponded well with constant for the 'slow' component discernible in ICYP association with cerebral cortical membranes at 37 degrees C. The maximum binding values were 63, 29 and 5.6 fM/mg membrane protein in cortical, hypothalamic and anterior pituitary membrane fractions, respectively. Evaluation of the beta-adrenergic receptor subtypes using 4 selective competitors indicated an average 19% content of the beta 2-subtype in cortical membranes, while in hypothalamic membranes 47% of the receptors could be assigned to that subtype. In the anterior pituitary as well as in the cerebellum, the receptors were predominantly of beta 2-subtype. These findings are discussed in terms of possible physiological functions of beta-receptors in these tissues, including the regulation of the release of pituitary hormones. PMID- 6299468 TI - Tetrodotoxin-sensitive ion channels characterized in glial and neuronal cells from rat brain. AB - Ion channels were studied in primary neuronal and in primary glial cultures from rat brain by measuring the uptake of guanidinium, an ion that can permeate the Na+ channel. Neuronal cells exhibit a veratridine-stimulated (EC50 30 microM) guanidinium uptake, which is blocked by tetrodotoxin (IC50 30nM). This demonstrates the presence of a voltage-dependent Na+ channel. In glial cells veratridine + scorpion toxin, but not veratridine or scorpion toxin alone can stimulate a tetrodotoxin-sensitive ion uptake, thus indicating a 'silent' Na+ channel in the glial cells. Phentolamine, propranolol and various local anesthetic drugs (e.g. tetracaine, dibucaine) blocked the two different kinds of Na+ channels in the two cell populations investigated. PMID- 6299467 TI - Cellular mechanisms underlying the inhibitory surround of penicillin epileptogenic foci. AB - Penicillin applied locally to the surface of in vivo hippocampus or neocortex is known to produce not only periodic interictal spikes within a focus, but also altered cellular activities around the focus. In the zone adjacent to a hippocampal focus, for example, Dichter and Spencer recorded hyperpolarization depolarizing burst-hyperpolarization sequences, while further from the focus they recorded only hyperpolarizations (presumably IPSPs). We use here our previously developed model of interictal spike generation to show that these observations can be explained by the following two assumptions: (1) there is a gradient of effectiveness of synaptic inhibition (small within the focus and increasing with distance from the focus) caused by the gradient of penicillin concentration; and (2) the radius for recurrent inhibition is larger than the radius for recurrent excitation. Our model best fits the experimental data if we assume further that recurrent excitation extends from any one small cortical region to only some--not all--of its neighboring regions. PMID- 6299469 TI - Characterization and partial purification of neuroblastoma growth inhibitory factor from the culture medium of glioblasts. AB - Neuroblastoma growth inhibitory factor (NGIF) exists in the conditioned medium of normal rat glioblasts. When neuroblastoma cells (Neuro2a, NS-20Y, and N1E-115) were cultured in the presence of the factor, the cell growth rates and DNA synthesis were markedly inhibited and the morphological differentiation including neural process formation was induced. However, the factor neither altered the growth rate nor the morphology of non-neuronal cells such as glial cell lines (C6 and 354A) or fibroblast (3T3). The molecular weight of the factor was estimated to be 75,000 Mr by gel filtration with Bio-Gel P-200, and the isoelectric point was 5.8. The factor was devoid of esteropeptidase activity, and susceptible to protease and thermal treatment. The growth inhibitory action of the factor was unrelated to the intracellular contents of cyclic AMP and GMP. The ability of NGIF to suppress preferentially the neural growth suggests its regulatory role in normal brain development. PMID- 6299470 TI - Age related changes of enkephalin in rat spinal cord. AB - Met-enkephalin immunoreactive material content was found to be decreased in the cervical and thoracic segments of the spinal cord from rats aged 25 months as compared to young, 3-month-old, rats. No age-related variations were detectable at the lumbar level. Bio-Gel P 30 column chromatography of thoracic segment extracts indicates that the composition of the immunoreactive material is similar in the two age-groups investigated. At the thoracic level opiate receptor binding was also measured. Opiate receptor number is increased in the thoracic segments of the spinal cord from older rats. These age-related changes in immunoreactive Met-enkephalin content and opiate receptor number at spinal levels may contribute to determine an altered pain sensitivity during aging. PMID- 6299471 TI - Characteristics of [3H]prazosin binding to guinea pig brain alpha-adrenergic receptors: effects of estradiol and progesterone. AB - [3H]Prazosin was found to bind to sites on guinea pig brain membranes with alpha 1-adrenergic receptor characteristics. Treatment of ovariectomized guinea pigs with estradiol benzoate (EB) or EB followed by progesterone (P) did not affect [3H]prazosin binding to membranes from hypothalamus, preoptic area, amygdala or cerebral cortex. When added to the incubation mixture of the assay, estradiol, P, and other steroids decreased [3H]prazosin binding but only at high concentrations. These results do not support the idea that estrogen and progestin influence reproductive physiology through effects on brain alpha 1-receptors, although limitations of the methodology employed do not completely rule out this possibility. PMID- 6299472 TI - Regulation of opiate receptors in mouse brain: arcuate nuclear lesion induces receptor up-regulation and supersensitivity to opiates. AB - Lesion of the hypothalamic arcuate nucleus of the mouse by neonatal application of monosodium glutamate (MSG) increased the binding of [3H]dihydromorphine to membranes prepared from the midbrain. A saturation curve of [3H]dihydromorphine binding indicated that MSG increased the number of the opiate receptors. The MSG treated mice also exhibited an enhanced response to morphine and naltrexone regarding thermal pain sensitivity. The physiological implications of opiate receptors up-regulations upon arcuate nuclear lesion are discussed. PMID- 6299473 TI - Morphological and physiological studies on cultured nerve cells from guinea pigs infected with herpes simplex virus in vivo. AB - Adult guinea pigs were inoculated with type 2 herpes simplex virus (HSV) on the whole back skin, and nerve cells from the dorsal root ganglia (DRG) 6 days after infection were grown in tissue culture. Morphological and physiological properties of the cultured nerve cells from HSV-infected animals (HSV-NC) were compared to those of nerve cells of DRG from the control, non-virus infected animals (CON-NC). During the early period of the culture (0-4 days) growth of nerve cells and non-neuronal cells from the HSV infected animals was essentially the same as that from the control animals. HSV-antigen was present in only a small percentage of the HSV-NC by immunofluorescence (IF). Electrophysiological examination revealed that most of the HSV-NC exhibited a reduced capability of generating spikes, which was quantitatively described as a reduction in the Vmax of the Na spikes. This was interpreted as a reduction in the number of Na channel molecules in the plasma membrane of the HSV-NC, while the resting membrane properties of the same cells, such as the resting membrane potential, input resistance and capacitance, were essentially the same as those of the CON-NC. On 4-5 days in culture, some HSV-NC regained a full capability to generate Na spikes. We considered these nerve cells to have overcome the HSV infection and were now entering a latent period of HSV infection. About 1/3 of the HSV-NC still remained incapable of generating Na spikes. Viral antigen was detected in only 10% of the nerve cells. In the late stage of the culture, HSV infection in vitro was first observed as lysis of non-neuronal cells growing close to some HSV-NC. Nerve cells then started to lose their neurites and became spherical. Finally on 8-9 days most of the cells, including the nerve cells, were lost from the dishes as a result of a generalized infection of supporting cells, i.e. fibroblasts and Schwann cells. This study confirms our previous finding that the electrophysiological technique is much more sensitive than the IF method for the detection of HSV-infection in nerve cells. The results indicate that some nerve cells infected with HSV overcome the infection in vitro. This is interpreted as the entering of these nerve cells into a latent period of HSV infection in vitro. PMID- 6299475 TI - Development of beta-adrenergic receptors and their function in glia-neuron communication in cultured chick brain. AB - The beta-receptors of intact neuronal and glial cells of chick embryonic brain were studied via the specific binding of the beta-antagonist [3H]dihydro-L alprenolol ( [3H]DHA). Cells were cultivated in either highly homogeneous or mixed populations; the neuronal cells were also grown under the influence of glial conditioned medium (GCM) or 10(-11)-10(-10) M L-norepinephrine or L isoproterenol. The beta-receptors of both neuronal and glial cells proved to be positively cooperative (n = 2.5) and of high affinity, with a Kdapp of 98 and 44 pM, respectively. The Kdapp value was influenced only slightly by the different culture conditions. The receptor concentration was relatively low in the homogeneous neuronal and glial cultures (Bmax = 6.4 and 3.3 fmol/10(6) cells, respectively). It increased by a factor of 2-3 if development of the neuron-glia contacts in the culture was possible (mixed cultures). GCM and beta-agonists elevated the number of beta-receptors of the neuronal cells approximately 4-fold, even in the absence of glial cells. This receptor-number change was preceded by a well observable morphological differentiation. Both the morphological and the beta-receptor effects of L-norepinephrine were antagonized by L-propranolol. The beta-receptor number increased about 2-fold during a 10-day in vitro development, even in neuron-glia mixed cultures. PMID- 6299474 TI - Abundant, uniquely oriented endoplasmic reticulum in capillaries of the CNS: demonstration using reduced-osmium and glucose-6-phosphatase cytochemistry. AB - In the endothelial cells of capillaries in the rat CNS, we have observed abundant, circumferentially oriented, smooth, membrane-bound profiles, found just beneath, and parallel to, the abluminal plasmalemma. These structures are seen particularly well in tissue exposed to potassium ferricyanide-reduced OsO4. We studied these structures cytochemically, using glucose-6-phosphatase as a marker for endoplasmic reticulum, and acid phosphatase as a marker for the Golgi associated endoplasmic reticulum containing lysosomal enzymes (GERL). We found that they contained glucose-6-phosphate hydrolyzing activity but did not contain acid phosphatase activity. Comparable structures were not seen in the continuous capillaries of skeletal muscle. Based on their morphology and content of glucose 6-phosphate hydrolyzing activity, we conclude that these structures are uniquely oriented smooth endoplasmic reticulum, which is much more abundant in capillaries of the CNS than in other continuous capillaries. The function of this distinctive feature of the CNS capillary is not known. PMID- 6299476 TI - Regulation of choline acetyltransferase in primary cell cultures of spinal cord by neurotransmitter L-norepinephrine. AB - Neurotransmitter L-norepinephrine increased up to 8-fold the activity of choline acetyltransferase (CAT), the enzyme responsible for the synthesis of acetylcholine, in mouse spinal cord cells in culture grown for several days. The increase of CAT activity by L-norepinephrine was mediated by a beta-adrenergic receptor in the same manner as the response of intracellular cyclic AMP. Derivatives of cyclic AMP caused an increase of CAT activity to the level similar to that of L-norepinephrine. A cyclic AMP phosphodiesterase inhibitor, 3-isobutyl 1-methyl xanthine (IBMX), enhanced the elevation of CAT activity by L norepinephrine. These results indicate that L-norepinephrine stimulated the synthesis of CAT molecules via the action of cyclic AMP. The pretreatment of cells with 5-fluoro-2'-deoxyuridine (FdU) markedly diminished the numbers of satellite cells and, in parallel, the responses of CAT activity to L norepinephrine. The increase of cyclic AMP by L-norepinephrine was also reduced by pretreatment of the cells with FdU. In contrast, co-cultures of spinal cord with heart muscle markedly (30-fold) stimulated CAT activity both with and without pretreatment of FdU. The addition of L-norepinephrine and co-cultures with heart muscle showed an additive effect. These observations indicate that the stimulatory effect of L-norepinephrine on CAT activity is mostly, if not only, mediated via the interaction with satellite cells, and that the increase of CAT activity by L-norepinephrine is based on a mechanism different from that of co cultures with heart muscle cells. PMID- 6299477 TI - Early recovery of function after olfactory tract section correlated with reinnervation of olfactory tubercle. AB - Behavioral recovery and cortical reinnervation after early olfactory tract section were assessed in the infant golden hamster (Mesocricetus auratus). Hamster pups show strong thermotaxis at birth which declines abruptly after postnatal day (P) 8 in normal pups. Unilateral olfactory bulbectomy on P5 causes persistent thermotaxis through the second postnatal week. In this study, the bulb's output pathway, the lateral olfactory tract, was unilaterally severed on P5 and pup thermotaxis was tested through P15. Complete tract section, like bulbectomy, prolonged thermal responding beyond P8. In contrast to bulbectomy, however, some tract-sectioned pups showed recovery before P15 while others continued to show persistent thermotaxis throughout testing. The olfactory bulb projection was examined 10 days after tract section in order to determine whether recovery and persistent thermotaxis were associated with different patterns of cortical innervation. Eleven pups with complete transections showed recovery during the second week. In 10 of these pups, olfactory bulb fibers had penetrated the damaged region after surgery to reinnervate the olfactory tubercle. Three of these pups also exhibited some reinnervation of piriform cortex. The lesions of pups showing persistent thermotaxis were more severe, extending bilaterally or into deep cortical layers, and olfactory fibers had failed to reinnervate caudal terminal fields. All pups with olfactory tract sections showed extensive sprouting rostral to the cut, regardless of their behavioral profile. In no case had postlesion growth innervated the entorhinal or amygdaloid areas. Inhibition of thermotaxis was associated with reinnervation of the olfactory tubercle rather than more rostral, lateral or caudal olfactory cortex. We conclude that regrowth of olfactory tract fibers caudal to early transection is rapid and has functional consequences for early behavioral development. PMID- 6299478 TI - A [14C]2-deoxyglucose analysis of the functional neural pathways of the limbic forebrain in the rat. III. The hippocampal formation. PMID- 6299479 TI - [Effect of passive immunization with an antiCRF antiserum on the secretion of ACTH in rats]. AB - A peptide with 41-residue having CRF activity both in vivo and in vitro has recently been isolated from ovine hypothalami (41-CRF). Passive immunization of Sprague-Dawley Rats with an antiserum against this peptide is followed by a significant decrease in plasma ACTH and cortisosterone levels under basal conditions as well as after ether-stress and adrenalectomy. These data demonstrate that 41-CRF plays a major role in the physiological regulation of ACTH secretion. PMID- 6299480 TI - [The frequency of chromosomal aberrations and sister chromatid exchange in the peripheral lymphocytes of patients with trophoblastic tumors]. PMID- 6299481 TI - Health and safety in the workplace. Part 4. California TB rates rising. PMID- 6299483 TI - Neonatal diarrhea of pigs in Quebec: infectious causes of significant outbreaks. AB - To evaluate the relative importance of the various enteropathogens causing neonatal diarrhea in Quebec farrowing operations, observations were made on 749 diarrheic pigs from 325 outbreaks of diarrhea. They were one to 15 days of age, and were obtained alive for necropsy generally within 48 hours of the onset of diarrhea. Some pigs were from severe, explosive outbreaks of diarrhea with high morbidity and mortality rates, while others were from herds with chronic neonatal diarrhea with lower morbidity and mortality rates. A combination of bacteriological, virological and histological methods were used to study the pigs. Viruses were incriminated in 60%, bacteria in 23% and coccidia in 15.3% of the 325 diarrhea outbreaks. Transmissible gastroenteritis virus was by far the most common enteropathogen with a prevalence of 52%; rotavirus was implicated in 9.2% of the outbreaks while adenovirus was incriminated in 0.30% of the outbreaks. Enterotoxigenic Escherichia coli were involved in 22.4% of the cases while Clostridium perfringens type C was an occasional finding. Coccidia involved in our herds were identified as Isospora suis. The disease was attributed to infection with a single etiologic agent in 590 diarrheic pigs (78%) while combinations of agents were present in only 90 (12%). The age-specific occurrence of the various enteropathogens was evaluated. Transmissible gastroenteritis virus was the most common enteropathogen in all age groups. Colibacillosis was common in pigs which became diarrheic under five days of age; in this age group, the enterotoxigenic E. coli were frequently found alone, but were usually combined with other agents in older pigs. The prevalence of coccidia was high in pigs which became diarrheic between five and 15 days of age. Rotavirus infection was common in diarrheic pigs older than ten days of age. Although individual baby pigs were commonly infected with a single enteropathogen, it was very common to see more than one agent involved in an outbreak of diarrhea, particularly when pigs of different ages were affected. Observations on the occurrence of the enteropathogens according to the seasons were also made. Occurrence of transmissible gastroenteritis was throughout the year with the highest prevalence during the fall, winter and spring months. Colibacillosis and coccidiosis were more common in the summer, fall and early winter months with the lowest prevalence in the spring months. PMID- 6299482 TI - Hyper(ADP-ribosyl)ation of histone H1. AB - Nucleosomal chains of various repeat unit lengths were generated by a mild micrococcal nuclease digestion of purified pancreatic nuclei. Maximal nucleosome associated poly(ADP-ribose) polymerase activity was recovered in trimeric to tetrameric chromatin fragments, after which the enzyme activity gradually decreased and stabilized towards oligomeric periodicities of 11 to 16 nucleosomes. Electrophoresis of [32P]ADP-ribosylated histones on first-dimension acid-urea or acid-urea-Triton gels and on second-dimension acid--urea- cetyltriammonium bromide gels revealed that, of all histones, only histone H1 could be significantly poly(ADP-ribosyl)ated while only minimal modification could be recovered with histone H1(0). Furthermore, the extent of ADP ribosylation present on pancreatic histone H1 is shown to proportionally retard this protein's electrophoretic mobility in all gel systems and to consist of a distinct series of at least 12 modification intermediates which can be evidenced, in nuclei or nucleosomes, and fully recovered along with histone H1 upon its selective extraction with 5% perchloric acid. The generation of these increasingly ADP-ribosylated forms of histone H1 is also demonstrated to be time dependent and the more complex ADP-ribosylated forms of this histone are favored at high NAD+ concentrations. Moreover, the electrophoretic mobilities of all intermediates are unaffected by the presence of the nonionic detergent Triton X 100. PMID- 6299484 TI - Detection of infectious bovine rhinotracheitis and bovine viral diarrhea viruses in the nasal epithelial cells by the direct immunofluorescence technique. AB - Nasal epithelial cells were collected by cotton swabs for the diagnosis in experimental and field cases of infectious bovine rhinotracheitis and field cases of bovine viral diarrhea in calves. A portion of the cells was washed twice in phosphate buffered saline and a 25 microL drop was placed on microscope slides. The cells were dried, fixed and stained according to the direct fluorescent antibody technique. Another portion of the same specimen was inoculated onto primary bovine skin cell cultures for virus isolation. In the experimental studies for infectious bovine rhinotracheitis, 29/35 specimens were positive by fluorescent antibody technique and 32/35 by cell culture and in the field cases, 22/119 were positive by fluorescent antibody technique and 19/119 by cell culture. In the field cases of bovine viral diarrhea, 28/69 samples were positive by fluorescent antibody technique and 14/69 by cell culture. When fluorescent antibody technique was performed on inoculated cell cultures a total of 24/69 specimens were positive for bovine viral diarrhea. The sensitivity of fluorescent antibody technique was thus comparable to that of cell culture method for infectious bovine rhinotracheitis and bovine viral diarrhea. PMID- 6299485 TI - Effect of viral dose on experimental pneumonia caused by aerosol exposure of calves to bovine herpesvirus 1 and Pasteurella haemolytica. AB - The effect of various aerosol doses of bovine herpesvirus 1, followed four days later by aerosol exposure to a constant level of Pasteurella haemolytica, was studied in 16 crossbred Hereford range calves. A Collision nebulizer was used to generate aerosols from virus suspensions with concentrations of 10(8.2) (high), 10(5.2) (moderate) or 10(2.2) (low) TCID50/mL. The bacterial suspension contained 10(7) colony forming units/mL. Control calves exposed only to P. haemolytica developed no pulmonary lesions. Calves in the low, moderate and high virus exposure groups developed lobular areas of atelectasis; in addition, one calf in the moderate and all four in the high virus exposure group developed fibrinous pneumonia. One of the latter calves died. The 50% effective dose for fibrinous pneumonia under these experimental conditions was 10(6.0) TCID50 bovine herpesvirus 1/mL of suspension in the nebulizer reservoir, and approximately 10(4.0) infectious units inhaled per calf. PMID- 6299486 TI - Infectious center assay of intracellular virus and infective virus titer for equine mononuclear cells infected in vivo and in vitro with equine herpesviruses. AB - A novel, simple method of infectious center assay was developed to detect and quantitate the intracellular existence of equine herpesvirus 1 and equine herpesvirus 2 in peripheral blood mononuclear cells infected in vivo and in vitro with the viruses by cocultivation of these cells with a permissive equine cell culture. The infectious center titers were correlated with the infectious virus titers. In vivo equine herpesvirus 1-infected mononuclear cells obtained from ponies experimentally infected with the virus and equine herpesvirus 2-infected mononuclear cells obtained from selected naturally infected ponies with the virus gave by infectious center assay a mean value of 67 infectious center/2 x 10(6) cells as a peak titer on day 4 postinfection and 26 infectious center/2 x 10(6) cells for equine herpesvirus 1 and equine herpesvirus 2 respectively. The mononuclear cells, in both cases, did not contain detectable infectious virus, but the infectious virus was detected from the respective cells when they were cultured in the presence of mitogen. The equine herpesvirus 1 infected mononuclear cells in culture gave a mean count of 8.05 x 10(2) infectious center/2 x 10(6) cells/mL and contained 1.08 x 10(4) plague assay/mL of infectious virus. Similarly the equine herpesvirus 2 infected mononuclear cells in culture gave a mean count of 7.1 x 10(1) infectious center/2 x 10(6) cells/mL and contained <10(1) tissue culture infective dose(50)/mL of infectious virus. Mononuclear cells infected in vitro with equine herpesvirus 1 gave a mean count of 9.3 x 10(4) infectious center/2 x 10(6) cells/mL and contained 5.75 x 10(3) plaque assay/mL of infectius virus. Culturing these cells in the presence of mitogen gave a mean count of 5.5 x 10(3) infectious center/2 x 10(6) cells/mL and contained 9 x 10(3) plague assay/mL of infectious virus. A correlation between infectious center assay and infectious virus assay is discussed. PMID- 6299487 TI - Dietary influences on excretory pathways and tissue residues of zearalenone and zearalenols in the rat. AB - Experiments were conducted with male weanling rats to determine the effect of high dietary protein (40% casein), alfalfa (25%), protein + alfalfa (25% casein + 25% alfalfa), or anion-exchange resin (5%) on the relative proportions of free and conjugated zearalenone and zearalenols excreted in urine and feces following a single oral dose of zearalenone. About 90% of the dose was excreted in feces after 48 h while the remaining 10% was excreted in urine. Zearalenone and metabolites were excreted mainly in free form with conjugates being found only in urine. Rats fed high-protein diets excreted more free zearalenone and alpha zearalenol in urine than did controls (16.3% casein). The feeding of protein + alfalfa also resulted in increased urinary losses of free zearalenone. A reduction in urinary losses of conjugated zearalenone and alpha-zearalenol was seen when 5% anion-exchange resin was fed. Fecal excretion of zearalenone and alpha- and beta-zearalenol increased when alfalfa + protein was fed. Residual zearalenone and zearalenols were measured in liver and kidney 18 h after dosing. All treatments except high protein reduced residues of zearalenone and alpha zearalenol in liver when compared with controls. Renal residues of zearalenone were lowered only by supplements of protein + alfalfa or anion-exchange resin. Each of the treatments fed has been shown to reduce zearalenone toxicosis in rats, but it was concluded that each does so by a different mechanism. PMID- 6299488 TI - Some functional and morphological characteristics of an acutely dispersed purified cell suspension of rat lactotrophs prepared with Percoll. AB - A cell suspension containing more than 90% lactotrophs can be prepared from enzymically dispersed adenohypophyses obtained from male rats pretreated with estradiol. The lactotrophs are separated from the mixed cell population by centrifugation on a discontinuous density gradient prepared from a commercial preparation of colloidal silica (Percoll, Pharmacia). The method allows isopycnic separation of these delicate cells under very mild conditions; normal ionic strength and normal pH were maintained throughout the gradient, centrifugal acceleration did not exceed 1600 X g, and all procedures were done at room temperature. Histological verification that at least 90% of the cells were lactotrophs was done using specific immunoperoxidase staining. The functional capability of the lactotrophs was established by measuring the dose--response to the dopamine agonist bromocriptine and to thyrotropin-releasing hormone (TRH). Bromocriptine decreased spontaneous release in a dose-related way over the concentration range of 10(-10) to 10(-8) M. TRH, which causes an in vivo release of prolactin (PRL) in estrogen-primed rats, produced a dose-related increase in the release of PRL over the concentration range of 3 X 10(-10) to 3 X 10(-8) M after the high spontaneous release had been previously reduced by bromocriptine (3 X 10(-8) M). PMID- 6299489 TI - Effects of reduced PO2 on rapid-drive-induced hyperpolarization of diastolic transmembrane potential in feline cardiac Purkinje strands. AB - Using standard microelectrode techniques, we evaluated effects of diminished oxygen tension on the magnitude and time course of frequency dependent changes in maximum diastolic transmembrane potential (MDP) and on alteration of action potential duration (APD) in feline Purkinje fibers. MDP was recorded continuously during a control period (cycle length (CL) = 1000 ms), during a 5-min period of rapid drive (CL = 400 ms) and following return to pacing CL = 1000 ms. Rapid drive resulted in hyperpolarization of MDP from control value; and after return to pacing CL = 1000 ms. MDP gradually depolarized, eventually attaining a steady state value within +/- 0.5 mV of the control value. The difference between hyperpolarized MDP value and final steady-state value was designated VH, and the decline of MDP towards steady-state value approximated an exponential function (time constant = tau VH). Exposure to reduced PO2 (75 +/- 2.1 mmHg vs. control 473 +/- 39.1 mmHg) (1 mmHg = 133.322 Pa) resulted in reduction in the magnitude of VH (6.2 +/- 3.43 mV vs. 7.8 +/- 2.73 mV, mean +/- SD, p less than 0.005) and shortening of APD within 0-24 min, while measurable prolongation of tau VH (75 +/ 18.5 vs. 54 +/- 9.0 s, p less than 0.005) began at 25-49 min following onset of reduced PO2. These observations suggest that rate-related changes of MDP in cardiac tissues are oxygen dependent, and they support previously reported analagous observations in nerve which suggested that frequency dependent potential changes may in part reflect alterations of electrogenic Na-K pump activity. PMID- 6299490 TI - The effect of displacement current on the measurement of reversal potential in squid giant axon. AB - The measurement of the sodium reversal potential (Erev), as that potential where the early current reverses during voltage clamp, was found to exceed the true Erev by 4.1 +/- 2.4 mV (mean +/- SD) in squid giant axon. This error was found in both intact and internally perfused axons and is due to interference from the displacement current. This was shown by subtraction of the current records obtained before and after treatment with tetrodotoxin (TTX). The error in Erev is proportional to (Td/gNA)12 where Td is the time constant of the displacement current. PMID- 6299491 TI - Hyperkalemia, hyperglycemia, and plasma levels of cyclic AMP and cyclic GMP induced by portal vein injection of cyclic nucleotides and their butyryl derivatives into dogs. AB - The levels of serum potassium, blood glucose, and plasma adenosine cyclic 3':5' monophosphate (cAMP) and guanosine cyclic 3':5'-monophosphate (cGMP) were studied after the portal vein injection of cyclic nucleotides and their derivatives, (cAMP, cGMP, N6, O2'-dibutyryl adenosine 3':5'-monophosphate (DBcAMP), N6 monobutyryl adenosine cyclic 3':5'-monophosphate (NMBcAMP), and O2'-monobutyryl adenosine cyclic 3':5'-monophosphate (OMBcAMP), into dogs. Dose-related hyperglycemic responses were observed after the injection of DBcAMP (1-8 mg/kg). Transient and prominent hyperkalemia and hyperglycemia were caused by the injection of DBcAMP, NMBcAMP, and OMBcAMP (4 mg/kg). The hyperkalemic response was highest with NMBcAMP (1.22 mequiv./L), followed by OMBcAMP (0.64), DBcAMP (0.54), cGMP (0.47), and cAMP (0.41), whereas the hyperglycemic response was highest with NMBcAMP (146 mg/100 mL), followed by DBcAMP (93.6), OMBcAMP (77.1), and cAMP (56.0), and there was only a slight change with cGMP (28.4) compared with the control. The plasma level of cAMP was maximal with DBcAMP (1.92 nmol/mL), followed by NMBcAMP (1.28) and OMBcAMP (0.76), whereas the plasma levels of cGMP showed no evident change, except that caused by DBcAMP (0.27). Of the cyclic nucleotides tested, NMBcAMP was found to be most potent in causing both hyperkalemia and hyperglycemia. Based on these results, possible correlations between hyperkalemia, hyperglycemia, and plasma levels of cAMP and cGMP are discussed. PMID- 6299492 TI - Rabbit atrial histamine receptors and cyclic AMP. AB - The present study confirms the presence of H2 receptors in rabbit right and left atrium by measuring changes in cyclic AMP. Histamine produced significant increases in cyclic AMP in both atria although the increases were less in left atria. Since H2 receptors are always associated with the adenylate cyclase- cyclic AMP system, the data provide further support for the suggestion that H2 receptors exit in rabbit right and left atria. PMID- 6299493 TI - A survey of enteric viruses in domestic sewage. AB - In this second study (1979-1981) of the viral content of sewage we have demonstrated the presence of poliovirus types 1, 2, and 3 in Laval and Montreal. Several strains of poliovirus types 2 and 3 were nonvaccinal. This is in contrast with our first study (1977-1978) in which only type 1 poliovirus isolates were nonvaccinal. Coxsackievirus types B-3, B-4, and B-5 and echovirus types 1, 7, and 11 were also isolated from sewage. Interestingly, these isolations coincided with reports of isolation of the same strains during the same period by diagnostic laboratories. Our method based on Vero and BSC-1 cell cultures for virus isolation and immune electron microscopy for identification permitted the recovery not only of several strains of enteroviruses but also of some adenoviruses and reoviruses. PMID- 6299494 TI - Sodium intoxication caused by use of baking soda as a home remedy. PMID- 6299495 TI - Screening for occult lung cancer. AB - A pilot screening program for the early detection of lung cancer was carried out in Saskatchewan in 1968 using chest roentgenography and cytologic examination of sputum samples. The yield from 23 000 men aged 40 years and over was only 10 cases. Nine of the men had advanced disease. One had occult lung cancer. A period of 31 months elapsed between the discovery of malignant cells in this patient's sputum and roentgenographic localization of the tumour. Following pneumonectomy he has survived with no discernible residual or metastatic tumour for 12 years. The morphologic changes in the resected lung provided a basis for discussing the preclinical phase of squamous cancer of the lung, the treatment of occult cancer and multicentric primary pulmonary tumours. The survey would have been more successful with a narrower target group and more frequent examination. PMID- 6299496 TI - The discovery of Burkitt's lymphoma. AB - An account is given of the steps that led to the recognition of the tumor that became known as Burkitt's lymphoma. This is followed by a description of the methods employed in the subsequent epidemiologic studies, the manner in which successive hypotheses of etiology were erected and demolished, and the discovery by Epstein and his colleagues of the Epstein-Barr virus. The impact that this tumor has had on aspects of cancer research is considered. PMID- 6299497 TI - The epidermal growth factor (EGF). AB - This review summarizes briefly some of the historical aspects of the discovery, isolation, and mode of action of epidermal growth factor (EGF). Also considered are some of the key experiments which relate the biochemical mechanisms of EGF action to the mechanism involved in oncogenic transformation by the retroviruses. PMID- 6299498 TI - Intramammary lymph nodes. AB - Radiographic, gross, and histopathologic studies on 158 whole breasts with primary operable carcinoma revealed intramammary lymph nodes in 28%, and of these breasts, 10% contained a metastatic deposit of carcinoma. Cancerous and noncancerous nodes were found in all quadrants of the breast with the positive ones being in the same quadrant as the carcinoma only 50% of the time. There was no demonstrable connection with the usual lymphatic drainage of the breast. With Stage II carcinoma, positive intramammary lymph nodes had no direct effect on prognosis, merely representing advanced disease and indicating a greater likelihood of axillary metastatic disease. There was a trend toward poorer prognosis in Stage I lesions with positive intramammary lymph nodes. This may indicate the Stage I carcinomas that have a similar prognosis as Stage II tumors. Conceivably, a Stage Ia, positive intramammary lymph node(s) but normal axillary lymph nodes, could be defined and used. PMID- 6299499 TI - The fine structure of a virilizing human granulosa-theca cell tumor. Observations on the nature of the hormone producing cell. AB - The ultrastructure of a virilizing granulosa-theca cell tumor is reported. Although the histologic appearance is characteristic of a granulosa-theca tumor, several tumor cells contain crystalloids of Reinke which have not been associated with sex-cord-stromal tumors, but which are often present in ovarian hilus cells and Leydig cells. The steroidogenic cell in this case displays many fine structural features common to mammalian steroidogenic cells, namely abundant smooth endoplasmic reticulum, a dispersed Golgi, mitochondria with tubular cristae, and lipid droplets. Other features of this cell, which contains crystalloids of Reinke, more closely resemble those of a theca-lutein cell than a granulose-lutein or hilus cell and support a stromal origin. However, this tumor illustrates the striking overlap in morphologic features and function among granulosa, theca, hilus, and Leydig cells. PMID- 6299500 TI - Carcinoid tumor of the gallbladder associated with adenocarcinoma. AB - A case of carcinoid tumor of the gallbladder associated with adenocarcinoma in a 56-year-old man is reported and a review of the literature is made. The tumor was a polypoid mass with a size of 5.5 X 4.0 X 2.8 cm. Histologically, the tumor showed carcinoid and adenocarcinoma with areas of mucous change. Tumor cells containing argyrophil granules were observed in both carcinoidal and adenocarcinomatous areas, but no argentaffin granules were detected in either of the neoplastic areas. Some of the tumor cells had both argyrophil granules and mucin in the same cytoplasm. The electron microscopic study revealed several tumor cells containing neurosecretory granules; however, no clinical signs of hormonal activities of the tumor were observed. The patient died of generalized bone metastases 16 months after surgery. This appears to be the second case of composite tumor of the gallbladder. PMID- 6299501 TI - Multiple small cell carcinomas of the esophagus. AB - Small cell carcinoma of the esophagus is extremely rare, and usually has a rapidly progressive course. This article reports a patient with synchronous multiple small cell carcinomas of the esophagus who had a prompt and complete (although brief) remission following treatment with a combination chemotherapy program used in patients with small cell lung cancers. Recognition of this entity is important, since appropriate treatment with multiagent chemotherapy has a major impact on prognosis. PMID- 6299502 TI - Androgen related primary hepatic tumors in non-Fanconi patients. AB - Three cases of androgen related primary hepatic tumor in non-Fanconi patients are described in whom detailed information is available concerning histologic changes and clinical course over long periods. Two patients presented with hepatic rupture and hemoperitoneum requiring surgical intervention. In one case histologic assessment showed a hepatic adenoma which has almost completely regressed over four years since androgen withdrawal. In two patients there was a degree of nuclear pleomorphism not seen in 'spontaneous' adenomas and following androgen withdrawal there has been no evidence of progression or of metastases although the tumor has not regressed over a follow-up period of two years. PMID- 6299503 TI - Protons or megavoltage X-rays as boost therapy for patients irradiated for localized prostatic carcinoma. An early phase I/II comparison. AB - A total of 180 patients with carcinoma of the prostate limited to the pelvis were treated with one of two external beam irradiation techniques between 1972 and 1979. One hundred and sixteen patients were treated with conventional pelvic megavoltage x-ray therapy. Sixty-four patients were treated with combined pelvic x-ray therapy plus a perineal proton beam boost to a carefully defined prostatic tumor volume. A 160 MeV proton beam has been modified to irradiate patients with localized tumors by using conventional treatment schedules. This proton beam has the physical advantage over megavoltage x-rays of reducing the dose to normal tissues adjacent to the tumor volume. By using the proton beam boost we have delivered an increased prostatic tumor dose of 500 to 700 cGy without increasing treatment morbidity at all. The two groups are actuarially analyzed for patient survival, disease-free survival and local recurrence-free survival, and thus far, no significant differences have been noted. Because of the minimal complications observed in the proton group despite a 10% increase in dose, a randomized clinical trial comparing these two treatment techniques is studied. PMID- 6299504 TI - The origin of germ cell tumors of the testis. AB - Two hundred and eight cases of germ cell tumors of the testis were thoroughly studied. Fifty-one (24.5%) were embryonal carcinoma, 115 (55.3%) were seminoma, and 42 (20.2%) contained both embryonal carcinoma and seminoma. The average age of the patients with embryonal carcinoma, seminoma, and combined tumors was 27.6, 38.3, and 30.3 years. Furthermore, in the patients with combined tumors, those having a predominance of embryonal carcinoma had an average age of 28.3 years, while those having a predominance of seminoma had an average age of 33.4 years. This study supports the concept that the malignant potential of germ cells in younger patients is embryonal carcinoma, the malignant potential of germ cells in older patients is seminoma, and that in intermediate-aged patients the germ cells are capable of developing varying amounts of embryonal carcinoma and seminoma in the same tumor. PMID- 6299505 TI - Pancreatic islet cell carcinoma associated with multiple hormone secretion and pancytopenia. Evidence of a serum factor suppressing hematopoiesis. AB - A case of metastatic islet cell carcinoma of the pancreas associated with the production of multiple polypeptide hormones (insulin, glucagon, and gastrin) is described. Three years prior to the histologic diagnosis the patient presented with a gastric ulcer and an androgen responsive pancytopenia with hypoplastic bone marrow. Discontinuation of androgen therapy resulted in relapse of pancytopenia. After the diagnosis of islet cell carcinoma of the pancreas was established, the patient was treated with 5-fluorouracil (5-FU) and streptozotocin and subsequently elevated serum polypeptide hormones returned toward normal levels. Concurrent with the normalization of peptide hormones another complete hematologic remission was achieved without use of androgens. Injection of the patient's serum into female rats produced a significant fall in leukocyte (P less than 0.02) and platelet counts (P less than 0.005), but no significant decrease in hematocrit. The clinical course and laboratory findings in this case suggest the presence of a previously undescribed serum factor released by an islet cell tumor capable of suppressing hematopoiesis. PMID- 6299506 TI - Partly luteinized theca cell tumor of the ovary. AB - The partly luteinized theca cell tumor is a variant of the theca cell tumor in which extensive foci of luteinization occur. This neoplasm belongs to a set of tumors, which includes the theca cell tumor, that is believed to be derived from mature ovarian stroma. This tumor can be classified as a tumor of specialized gonadal stroma of "ovarian cell type" intermediate between the theca cell tumor and the stromal luteoma. Two patients were virilized, and one had evidence of endometrial hyperplasia, whereas in the fourth no endocrine function was evident. None of the patients were pregnant at the time of discovery of the tumor. On gross examination these tumors consisted of a mixture of firm gray-white and yellow tissue. The presence of multiple yellow nodules in two tumors distinguished it from the usual theca cell tumor. A third tumor was distinctly lobulated. The tumor in only one of the cases arose in a background of ovarian stromal hyperplasia. Although the number of cases reported up to now is small and variability exists, the tumors tend to occur in patients in the early reproductive age group, often produce significant quantities of steroid hormones, and the clinical course has been benign. PMID- 6299507 TI - Primary anaplastic small cell (oat cell) carcinoma of the larynx. Review of the literature and report of 18 cases. AB - Anaplastic small cell (oat cell) carcinoma is a neoplasm commonly arising in the lungs. However, it may also occur, though rarely, in the larynx. A series of 43 cases is presented (ten cases from the Armed Forces Institute of Pathology, eight from the Department of Otolaryngology of the Padua University, and 25 from the literature). The tumor often presents in the sixth and seventh decades of life and appears to be highly aggressive, and metastases develop early. The most common presenting symptom is hoarseness. As in pulmonary small cell carcinoma, prognosis is poor and does not seem to depend upon therapeutic modalities, tumor location or the extent of initial local disease. The tumor seems to derive from the Kulchitsky cell present not only in the bronchial mucosa but also in the laryngeal lining. Like pulmonary anaplastic small cell carcinoma, small cell carcinoma of the larynx should be treated with systemic chemotherapy and radiotherapy. The association of small cell carcinoma with squamous carcinoma of the larynx is also reported and problems connected with the histogenesis of this mixed tumor are discussed. PMID- 6299508 TI - Chromosome changes in rat embryo cell lines transformed by temperature-sensitive mutants and sheared DNA of herpes simplex virus. AB - The chromosomes of six rat embryo cell lines transformed with herpes simplex virus (HSV) temperature-sensitive (ts) mutants were examined at different passages of in vitro cultivation. Two cell lines were predominantly diploid, one cell line was hyperdiploid, one cell line was pseudodiploid, and two cell lines were hypotetraploid. In near-diploid cell lines chromosome No. 9 was most frequently involved in chromosome changes. All three cell lines derived from tumors obtained after one transplantation of HSV-transformed cells into baby rats were pseudodiploid, but each had different marker chromosomes. Chromosome No. 15 was involved in the formation of two out of four marker chromosomes. Four cell lines derived from tumors developing after two and three transplantations were hypodiploid and showed large chromosome variation. The occurrence of 25 marker chromosomes in three tumor-derived cell lines resulted in gains in parts from chromosomes No. 2, 6, and 7. One marker chromosome had a homogeneously faintly stained region. Chromosomes No. 2, 3, 7, and 12 were more frequently involved in the formation of marker chromosomes. No chromosome change was found to be specifically associated with HSV-induced transformation of rat cells, but chromosome changes in tumor-derived cell lines may provide selective advantage for survival and autonomous growth in the host animal. PMID- 6299509 TI - Fibrous histiocytoma cell line: chromosome studies of mouse and man. AB - A human cell line isolated from a lung metastasis of a malignant fibrous histiocytoma was studied chromosomally. The cell line had a modal chromosome number of 59 with multiple numerical and morphologic chromosome changes and marker chromosomes. A putative clone from this cell line had a modal number of 41 with exclusively acrocentric chromosomes and was clearly not human but mouse in origin. PMID- 6299510 TI - Mutagenic virus replication in human tumor cells. AB - Herpes simplex virus was grown in different lines of human tumor and normal cells. The progeny virus was assayed for resistance to iododeoxycytidine, an indicator of a forward mutation in the virus genome. Virus grown in cells from 4 of 5 tumor lines demonstrated greater fractions mutated to iododeoxycytidine resistance than did virus grown in 7 normal human skin cell lines. The data indicate that some lines of human tumor cells modify the herpesvirus replication process, making it more mutagenic. In 2 cases of osteosarcoma patients, normal skin fibroblasts of the patients yielded normal levels of mutagenesis, while their tumor cells gave enhanced mutagenesis. PMID- 6299511 TI - Modification of herpes 2-transformed cell-induced tumors in mice fed different sources of protein, fat and carbohydrate. AB - The effects of different sources of protein (milk, soy, wheat, fish and beef), fat (corn oil and butter), and carbohydrate (dextrin and sucrose) on tumor development and on spleen characteristics were investigated in BALB/c mice injected subcutaneously with 5 X 10(5) herpes simplex virus Type 2-transformed cells (H238 cells). Low or high levels of protein and fat were used. Several weeks post-injection results indicated that a high level of fat significantly enhanced tumor incidence. A high fat level was also associated with a lower spleen weight and a smaller proportion of mature granulocytes in the spleen. Butter, compared to corn oil, significantly restricted tumor volume. Among the most highly significant findings was the low tumor incidence in mice fed protein from either a milk or a fish source. PMID- 6299512 TI - Different weekly changes in immune response in virus-transformed cell-injected mice fed two different diets. AB - Changes in tumor development and in certain immune responses were investigated at 7-weekly intervals after subcutaneous injection of 5 X 10(5) herpes simplex virus Type 2-transformed cells (H238 cells) into male BALB/c mice fed 2 different diets. One diet contained 11% casein and 5% fat while the other had 11% supplemented wheat gluten and 30% fat. Weanling mice (140/group) were fed one or the other of the diets for 12 weeks before injection and subsequent testing of 15 injected and 5 non-injected mice from each diet group each week. In mice fed the low-fat diet containing casein both tumor incidence and tumor volume were significantly lower (P less than or equal to 0.05) than in the group fed the 30% fat diet containing supplemented wheat protein. The casein-fed mice also had less splenomegaly and a higher proportion of mature lymphocytes in the spleen during tumor growth. The proliferative capability of the spleen cells after phytohemagglutinin stimulation was enhanced 2 weeks after H238 cell injection only in the casein-fed mice. PMID- 6299513 TI - Monoclonal antibody for identification of human duct cell carcinoma of pancreas. AB - Monoclonal antibody to duct cell surface marker (HP-DU-1), produced by hybridoma, was shown to be specific to duct cell surface antigen when tested against normal adult and 12-20-week-old fetal human pancreases. There were no common HLA antigens for locus A, B, C and DR among the 10 pancreas donors; nor did HP-DU-1 reveal any affinity for lymphocytes of donors whose pancreases were used as antigen for immunization of BALB/c mice. Seven out of 7 human pancreas adenocarcinomas including a poorly differentiated human pancreatic tumor expressed surface determinants detectable by HP-DU-1. Cell lines CAPAN-1 and CAPAN-2, derived from human pancreatic adenocarcinoma, also revealed the same antigenic determinants. PMID- 6299514 TI - Protection against hydroxyurea-induced cytotoxic effects in L5178Y cells by free radical scavengers. AB - Exposure of L5178Y cells in culture to 1 mM hydroxyurea (HU) for 3 h followed by 24 h incubation in an HU-free medium induced an abnormal enlargement of about 40% of the cells in the population and post-treatment reduction of DNA synthesis in comparison with control cells. These effects were used to examine the protection afforded by free radical scavengers against HU-induced cytotoxicity. It has been found that with careful choice of conditions (suitable concentration of the protective agent, pretreatment of cells) substantial protective effect of alpha tocopherol, sodium benzoate, acetylsalicylic acid, catalase, peroxidase or superoxide dismutase can be achieved. PMID- 6299515 TI - Effects of chrysotile asbestos on lung and pleural carcinogenesis of N-bis(2 hydroxypropyl)nitrosamine in rats. AB - The effects of chrysotile asbestos on lung and pleural carcinogenesis by N-bis(2 hydroxypropyl)nitrosamine (DHPN) in male Wistar rats were studied. Chrysotile, 30 mg per rat, was injected into the left pleural cavity and 3 g/kg body wt. DHPN was injected once into the abdominal cavity. Lung tumors (adenoma, adenocarcinoma, squamous cell carcinoma, and combined carcinoma) occurred at the highest incidence (100%). Adenocarcinoma was seen in 4 of 11 (36%) rats killed at 35 weeks and in 6 of 12 (50%) rats killed at 52 weeks, squamous cell carcinoma occurred in 1 of 11 (9%) rats killed at 35 weeks and 3 of 12 (25%) rats killed at 52 weeks, and mixed carcinoma was seen in 1 of 12 (8%) rats killed at 52 weeks, which received chrysotile and DHPN. Adenocarcinoma was seen in 9 of 11 (82%) rats which received DHPN only and killed at 52 weeks. Mesotheliomas were seen in 2 of 11 (18%) rats, killed at 35 weeks, and 3 of 12 (25%) rats, killed at 52 weeks, which received chrysotile and DHPN. Hyaline thickening of the pleura was seen in 100% of rats receiving chrysotile. Mesothelial cell hyperplasia and adenomatous and/or fibromatous growth of the mesothelium were seen in the pleura on both sides, ranging from 36% to 50% and 31% to 64% in rats receiving chrysotile and DHPN, respectively. Asbestos bodies were seen in the pleura on both sides and in the lung. PMID- 6299516 TI - Demonstration of type-R and type-C virus particles in hamster pancreatic adenocarcinomas. AB - Two transplantable solid tumors (CBP and PDP) and two continuous cultured cell lines (CBP-TC and PDP-TC) were established from nitrosamine-induced pancreatic ductal adenocarcinomas in inbred Syrian hamsters. Electron microscopic examination of both CBP and PDP solid tumors as well as CBP-TC cultured cells revealed the presence of type-R virus particles, which consisted of a 100 nm envelope containing a central 50 nm nucleoid with characteristic radiating 'spokes'. Type-R particles were abundant in cultured CBP-TC cells, moderately frequent in CBP solid tumors, and rare in PDP solid tumors; they were not present in PDP-TC cultured cells. The CBP and PDP solid tumors and cultured PDP-TC cells exhibited type-C viral particles, having a 150 nm envelope containing a 75 nm nucleoid. Mature, immature and budding forms of Type-C viruses were present and quite prominent in PDP tumors. Type-C particles were present but rare in CBP solid tumors and in PDP-TC cells. C-particles were not observed in cultured CBP TC cells. Examination of normal hamster pancreas, liver, duodenum, muscle and connective tissue failed to reveal evidence of type-R or type-C virus particles. It is important to recognize the presence of virus particles in hamster pancreatic carcinoma lines, since viruses could potentially affect biochemical or immunological studies on the carcinomas through the production of viral proteins that could be mistaken for tumor-associated antigens. PMID- 6299517 TI - Susceptibility of skin fibroblasts from patients with retinoblastoma to transformation by murine sarcoma virus. AB - Cultured skin fibroblasts from patients with retinoblastoma (RB) of different etiology have been studied for their susceptibility to transformation by murine sarcoma virus. The cells from patients with a deletion in chromosome 13 (13q-) and those from sporadic unilateral cases due to somatic mutation were as sensitive as normal cells. However, the cells from familial cases showed an extremely high sensitivity to transformation. Moreover, in familial cases the susceptibility was significantly higher in bilaterally affected patients than in unilateral cases. These findings suggest that the heritable RB gene is different from 13q- and its degree of expression is also manifest at the cellular level. PMID- 6299518 TI - Hormones of the pituitary-adrenal axis in rats bearing the Walker 256 carcinoma. AB - Male Sprague-Dawley rats (125-150 g) were implanted intramuscularly with the Walker 256 carcinoma. After 3, 5 or 7 days, tumor-bearing rats, along with controls, were killed and plasma levels of adrenocorticotropic hormone (ACTH), beta-endorphin and corticosterone were assessed. Plasma levels of all 3 hormones declined with time following tumor implantation. Plasma levels of ACTH and corticosterone were statistically significantly less than plasma levels of these same hormones in control rats by 7 days post-implantation. Levels of these hormones were reduced by 42% and 33%, respectively, relative to control levels by 7 days. beta-Endorphin levels declined much more rapidly following tumor implantation than did either of the other 2 hormones. beta-Endorphin was significantly decreased by 3 days post-implantation and by 7 days the plasma levels of this factor were 83% lower than in control rats. PMID- 6299519 TI - Calcitonin secretion by continuous cultures of small cell carcinoma of the lung: incidence and immunoheterogeneity studies. AB - A study was made of immunoreactive calcitonin (iCT) secretion by continuous cultures of small cell carcinoma of the lung (SCCL). Using an antiserum region specific for the midportion of the molecule, 9/12 cultures were found to secrete iCT. Gel filtration studies were performed on both supernatant fluid (SF) and cell pellet (CP) extract from a culture secreting high levels of iCT. Multiple iCT fractions were found in the SF with the major fraction being of high molecular weight (MW). In contrast, the CP had apparently monomeric CT as its principal iCT fraction. These studies demonstrate frequent iCT secretion by SCCL cultures and significant disparities between the iCT moieties found extra- and intracellularly. PMID- 6299520 TI - The influence of asbestos dust on the oncogenic transformation of C3H10T 1/2 cells. AB - The cell transforming ability of asbestos dust was investigated using C3H10T 1/2 murine fibroblasts. In a series of experiments both crocidolite and amosite caused no increase in the number of transformed foci over that seen in cultures from untreated cells. The dusts, were, however, capable of augmenting the oncogenic effect of benzo[a]pyrene (BP). This putative synergistic effect was evident when fibres and chemicals were added to cultures as simple mixtures and when BP was adsorbed to the surface of the fibres. PMID- 6299521 TI - Arrest of hormone-dependent mammary cancer growth in vivo and in vitro by cholera toxin. AB - Growth of 7,12-dimethylbenz(alpha)anthracene-induced mammary carcinoma in rat was arrested by daily s.c. injections of cholera toxin. At a dose of 2 micrograms/200 g rat, tumors regressed to 50% of their initial size within 2 weeks, and 85% of tumors regressed completely within 4 to 5 weeks. The same response to cholera toxin was observed with another hormone-dependent mammary tumor, MTW9, but not with the hormone-independent tumors, DMBA No. 1 and MT 13762. The latter result was consistent with the lack of response of these hormone-independent tumors to hormone removal (ovariectomy) or N6,O2'-dibutyryl cyclic adenosine 3':5' monophosphate treatment. The growth-inhibitory effect of cholera toxin was dose dependent, and upon cessation of treatment tumors resumed growth; after complete regression, however, tumors did not reappear until 6 months after termination of the treatment. An amount of cholera toxin as high as 10 micrograms/day/200-g rat s.c. injected over a 6-week period showed no systemic toxicity in the animals. The growth of human breast cancer cells (MCF-7) in culture was also inhibited by a daily supplement of cholera toxin. At a concentration of 100 ng/ml, the cell replication ceased completely within 2 days. The growth inhibitions, both in vivo and in vitro, were accompanied by marked increases in the cellular cyclic adenosine 3':5'-monophosphate content and type II cyclic adenosine 3':5' monophosphate-dependent protein kinase activity as well as a decrease of estrogen binding activity. Thus, extinction of mammary cancer can be achieved by cholera toxin, an agent that stimulates the intracellular cyclic adenosine 3':5' monophosphate system. PMID- 6299522 TI - Comparative effects of aplysiatoxin, debromoaplysiatoxin, and teleocidin on receptor binding and phospholipid metabolism. AB - We have compared the activities of aplysiatoxin and debromoaplysiatoxin, two polyacetate marine algae toxins, with teleocidin, a tumor-promoting indole alkaloid from Streptomyces, with respect to inhibition of specific binding of epidermal growth factor, and phorbol-12,13-dibutyrate to their respective receptors and ability to stimulate the release of radioactivity from cells prelabeled with choline or arachidonic acid. Although these compounds have chemical structures that are quite different from the phorbol esters, both aplysiatoxin and teleocidin are essentially equipotent with the potent tumor promoter 12-O-tetradecanoylphorbol-13-acetate in all four assays. The fact that aplysiatoxin and teleocidin inhibit phorbol-12,13-dibutyrate-receptor binding suggests that their biological activities are mediated by binding to the same receptors utilized by the phorbol esters. Debromoaplysiatoxin, a debrominated form of aplysiatoxin, is about 10-fold weaker than aplysiatoxin in inhibiting epidermal growth factor and phorbol-12,13-dibutyrate-receptor binding, but is equipotent with aplysiatoxin in stimulating the release of lipid metabolites from the prelabeled cells. The results are discussed in terms of possible heterogeneity of cellular receptors for this group of compounds. PMID- 6299523 TI - Effect of a 6-fluoro substituent on the metabolism and biological activity of benzo(a)pyrene. PMID- 6299524 TI - Involvement of macrophage-like cells in growth of tumors induced by avian sarcoma virus. PMID- 6299525 TI - Comparison of intra- and extracellular transforming growth factors from nontransformed and chemically transformed mouse embryo cells. AB - Multiple transforming growth factors (TGFs) have been isolated from the serum free conditioned media and acid-ethanol cell extracts of nontransformed AKR-2B and chemically transformed AKR-MCA mouse cells. The TGF activity was analyzed using Bio-Gel P-60 chromatography in 1 M acetic acid and tested for colony stimulating activity in soft agar using AKR-2B, AKR-2B (clone 84A), NRK, and AKR MCA cells as indicators. Both intracellular and extracellular TGF activity from AKR-MCA and AKR-2B cells show a major peak of AKR-2B and epidermal growth factor potentiated NRK colony-stimulating activity that coelute in the 13,000 +/- 2,000 molecular weight region. In the 24,000 +/- 7,000 molecular weight range, AKR-MCA cells produce intracellular and extracellular NRK colony-stimulating activity that is not potentiated by epidermal growth factor, while the intracellular and extracellular NRK colony-stimulating activity produced by AKR-2B cells requires added epidermal growth factor for colony formation. Also important in determining the growth and morphological characteristics of a cell line, besides the production of a TGF, is the ability of a cell to respond to TGF activity. We have shown that the transformed AKR-MCA cells form more colonies than AKR-2B cells in response to certain TGF activities. This suggests that the increased responsiveness of AKR-MCA cells to TGFs may be important in determining its phenotype. PMID- 6299526 TI - Comparison of spontaneous mutation rates of normal and chemically transformed human skin fibroblasts. PMID- 6299527 TI - Circulating levels of feline leukemia and sarcoma viruses and fibrosarcoma regression in persistently viremic cats. PMID- 6299528 TI - Binding of benzo(a)pyrene and (+/-)-7 beta,8 alpha-dihydroxy-9 alpha, 10 alpha epoxy-7,8,9, 10-tetrahydrobenzo(a)pyrene to histones. AB - AKR-2B mouse embryo cells were incubated for 24 hr with [3H]benzo(a)pyrene, and the histones were isolated and analyzed using one- and two-dimensional gel electrophoresis and autoradiography. The results revealed that (a) histones H1, H2A, and H3 incorporated significant amounts of label whereas little or no label was associated with histones H2B and H4 and (b) electrophoresis of the histones in the Triton:acid:urea gel system caused labeled histones to have a slower migration than did the corresponding unlabeled histones. Additional studies such as incubation of (+/-)-7 beta,8 alpha-[3H]dihydroxy-9 alpha,10 alpha-epoxy 7,8,9,10-tetrahydrobenzo(a)pyrene with nuclei resulted in radioactive labeling of histones H1, H2A, H2B, and H3 and of high-mobility-group proteins HMG1 and HMG2. The low levels of label associated with histone H4 in the whole-cell and nuclear studies were further investigated by incubating isolated histones with (+/-)-7 beta,8 alpha-[3H]dihydroxy-9 alpha,10 alpha-epoxy-7,8,9,10 tetrahydrobenzo(a)pyrene. Under these conditions, negligible amounts of radioactivity were associated with H4, while significant labeling of H1, H2A, H2B, and H3 and other nuclear proteins was observed. The results suggest that factors other than the presence of suitable nucleophilic acceptor sites on the histones may be necessary for carcinogen binding. PMID- 6299529 TI - Cytoskeleton-associated proteins of human lung cancer cells. PMID- 6299530 TI - Origin of phenotypic variation in clones of simian virus 40-transformed mouse embryo cells. PMID- 6299531 TI - Molecular identification of the TCA alloantigen as the transferrin receptor on T cells of a leukemic patient with serum from a monozygotic twin brother. AB - The transferrin receptor is encoded by a diallelic locus which had been described as the T-cell antigenic system TCA. Using a human lymphoblastoid T-cell line (HSB) which reportedly carries a transferrin receptor and which we showed to be TCA-2 positive by serology, it appeared that only a TCA-2 but not a TCA-1 specific alloantiserum had the capacity to precipitate a protein with a molecular weight of 94,000, which comigrated exactly with the Mr 94,000 protein precipitated by the rabbit anti-HSB and rabbit anti-SB antisera, as well as the transferrin receptor which was precipitated by a transferrin-antitransferrin complex. The positive sera and the latter complex were shown to cross-react in a binding assay. Conversely, a monospecific TCA-1 alloantiserum was shown to react with the Mr 94,000 protein of TCA-1-positive leukocyte suspensions. The allelomorphism of the transferrin receptor may be relevant for the eventual regulatory mechanism controlling cell proliferation, which would be essential at least for those cells that are supposed to play an immunological role like the TCA-positive cells which are described as T-cells probably involved in the process of immunosuppression. In addition, we found increasing amounts of autoantibodies with TCA-1 specificity in serum from a healthy individual (referred to as "donor" in this paper), whose brother at the same time suffered a chronic myeloid leukemia, probably associated with a retrovirus, and whose blood in contrast was shown to contain cytotoxic anti-TCB, from a diallelic locus expressed on theophylline-insensitive T-cells, and anti-B-cell activity. PMID- 6299532 TI - Leukocyte migration inhibition responses to MCF-7, murine mammary tumor virus, and Thomsen-Friedenreich antigen in a series of cancer patients. PMID- 6299533 TI - Chemical enhancement of viral transformation in Syrian hamster embryo cells by gaseous and volatile chlorinated methanes and ethanes. AB - Methods were developed for exposing cells in vitro to gases or vapors of volatilized organic liquids. Compounds were selected for their industrial importance, environmental impact, and suspected role in the etiology of some human cancers. Exposure chambers were designed for easy insertion of dishes of cultured cells and were equipped with inlet and outlet ports for introduction and purging of test gases. A gas delivery system utilizing a mass flow meter was used for the quantitative distribution of test gases into exposure chambers. For volatile compounds, appropriate volumes of cold (4 degrees) liquids in glass Petri dishes were quickly placed into chambers, the system sealed, and the compounds rapidly volatilized at 37 degrees. For exposure, the cells and chambers were placed in an incubator and rocked at a constant rate so that a portion of the cells was always in direct contact with the test gases or vapors. Known sample volumes were removed after various treatment times and test gas concentrations determined by standard gas chromatographic techniques. After exposure, the cells were removed and assayed for viability and increased sensitivity to viral transformation. Under these experimental conditions, the volatile liquids 1,1,1-trichloroethane, dichloromethane, chloroform, 1,2 dichloroethane, and 1,1-dichloroethane significantly enhanced transformation of Syrian hamster embryo cells by SA7 adenovirus, while acetone exerted no effect. The gases chloromethane and vinyl chloride were also active in this test system, while bromomethane, methane, and ethane were inactive. Incorporation of some of these compounds into liquid cell culture medium for cell treatment was either unsuccessful or produced only a weak enhancement response. Methodology is now available to evaluate volatile and gaseous carcinogens or mutagens and can be used to identify their mechanisms of action and the relative hazards of these agents to human health. PMID- 6299534 TI - Role of suppressor cells in feline leukemia virus-associated immunosuppression. AB - Protein from detergent-disrupted feline leukemia virus was found to suppress lymphocyte blastogenic responsiveness of peripheral blood mononuclear cells from normal cats. When suppressive doses of this protein were substituted for mitogen in the preactivation step of the concanavalin A-induced suppressor cell assay, significant suppression was observed in five of eleven cats tested. These results suggest that feline leukemia virus-associated immunosuppression may be mediated by a virus-induced suppressor cell. PMID- 6299535 TI - Transforming growth factors in solid human malignant neoplasms. AB - Surgically removed solid human benign and malignant neoplasms and nonneoplastic tissues were examined for the presence of transforming growth factors (TGFs). TGFs are polypeptide growth factor-like substances which cause the appearance of a reversible neoplastic phenotype in nontransformed, anchorage-dependent cells in culture, including the induction of the ability to grow while suspended in semisolid medium. Acid-ethanol extracts from adenocarcinomas of the breast, colon, kidney, and ovary; fibrosarcoma and leiomyosarcoma; Hodgkin's lymphoma; fibroadenoma of the breast; uterine leiomyoma; and nonneoplastic kidney and lung were found to cause growth in soft agar of both nontransformed mouse AKR-2B and rat NRK cells. This colony-stimulating activity, where tested, was heat and acid stable but was destroyed by trypsin and dithiothreitol treatment, indicating that the activity is due to a polypeptide with disulfide bonds. Extracts from several of the tumors provided sufficient material for purification by molecular sieve chromatography. Peaks of colony-stimulating activity from a Bio-Gel P-60 column eluted with 1 M acetic acid were detected in the M, 3,000 to 25,000 range with the apparent molecular weight varying depending on the type of tumor being studied and the indicator cells used. The data suggest that at least three TGFs are present in human tumors. Evidence is presented differentiating these TGFs into TGFa, which has selective activity for stimulating AKR-2B cells, and TGFn, which has selective activity for stimulating NRK cells. The NGFn activity was further subdivided into a TGFns fraction and TGFnl fraction, denoting small (less than 6,000) and large (12,000 to 20,00) apparent molecular weights, respectively. The TGFa and TGFnl activities were present in malignant and nonneoplastic (kidney and lung) tissue, whereas the TGFns activity predominated in benign neoplasms. These TGFs exhibited no competition with epidermal growth factor for binding to the epidermal growth factor receptor, and the TGFnl activity was potentiated by epidermal growth factor. PMID- 6299536 TI - Relationships between chemotaxis, chemotactic modulators, and cyclic nucleotide levels in tumor cells. AB - Like many freely moving cells, Walker 256 carcinosarcoma cells respond to chemotactic stimuli. Since cyclic nucleotides are involved in the chemotaxis of other cells, we have examined the action of several nucleosides and nucleotides as chemoattractants and as modulators of tumor cell movement. We have also studied the effect of chemoattractants and prostaglandins on intracellular cyclic nucleotide levels and the effect of prostaglandins as modulators of chemotaxis. Of the agents studied, only the cyclic nucleotides and prostaglandins were found to modulate cellular motility. Neither cyclic adenosine 3':5'-monophosphate (cAMP) nor cyclic guanosine 3':5'-monophosphate (cGMP) was a chemoattractant, but cGMP and N6,O2'-dibutyryl cyclic guanosine 3':5'-monophosphate at low concentrations (approximately 10(-10) M to 10(-8) M) enhanced chemotaxis by 80 +/ 15%, and both cAMP and N6,O2-dibutyryl cyclic adenosine 3':5'-monophosphate had an inhibitory effect at concentrations greater than 10(-6) M. Chemotaxis was suppressed by 21 to 100% in media depleted of Ca2+ and/or Mg2+, but in the presence of 10(-8) M cGMP, there was partial recovery of the chemotactic response. In response to chemotactic stimulation, there was a 28 to 60% rise in intracellular cAMP within 30 sec. This returned to basal levels within 2 min. Intracellular cGMP levels became elevated approximately 3- to 3.5-fold after this time. Incubation of cells with prostaglandins A1 and F2 alpha stimulated chemotaxis at lower concentrations (10(-7) and 10(-9) M, respectively) and resulted in elevation of cGMP, while incubation with prostaglandin E2 resulted in inhibition of chemotaxis and a rise in cAMP levels. PMID- 6299537 TI - Selective effect of Rous sarcoma virus src gene expression on contractile protein synthesis in chick embryo myotubes. AB - Myogenic precursor cells were infected with a temperature-sensitive mutant of Rous sarcoma virus and maintained at the permissive temperature for transformation. Following subculture, a population of cells was produced which failed to differentiate at the permissive temperature but produced a high proportion of myotubes in sister cultures shifted to the nonpermissive temperature. The myotube-containing cultures were further enriched for this cell type by the addition of 1-beta-D-arabinofuranosylcytosine to kill replicating mononucleated cells. This myotube population was suitable for testing the effect of viral-transforming gene expression in a postmitotic, terminally differentiated cell which expresses relatively low levels of the cellular homologue of the viral transforming gene and is resistant to infection by the virus. The consequence of shifting these Rous sarcoma virus-infected myotube cultures to the permissive temperature was assessed in terms of protein synthesis. The total rate of incorporation of exogenous radioactive leucine, supplied at a concentration which saturated the intracellular pool, was similar between cultures held at the two temperatures, suggesting that the absolute rate of total protein synthesis was not affected by expression of viral-transforming gene. In contrast, the rate of synthesis of eight proteins the expression of which is specific for myotubes was suppressed reversibly. The rate of synthesis of five other proteins which are not selectively concentrated in myotubes was either unaffected or stimulated. Thus. the expression of viral-transforming gene in myotubes leads to differential effects on developmentally regulated proteins without inducing several properties of the transformed state classically observed for fibroblastic cells. PMID- 6299538 TI - Effects of laser irradiation on hematoporphyrin-treated normal and transformed thyroid cells in culture. AB - Laser irradiation of tissues treated in vivo with the hematoporphyrin derivative (HPD) is known to result in a cytocidal effect, reportedly more pronounced in the tumor than in the surrounding normal tissues. In order to ascertain if this phenomenon had a clear cellular basis, it has been now reproduced in vitro in a model system consisting of normal and transformed cell lines. Epithelial rat thyroid cells were infected and transformed with a RNA oncogenic virus. Both the original (normal) and the viral-transformed (tumorigenic) cells were incubated with HPD and exposed to two types of laser irradiation: 631 nm, continuous wave; and 337.1 nm, pulsed. Under the conditions tested, the percentage survival of the transformed cells was found to be lower (up to approximately 3 times) than that of the normal cells. The cytocidal effect was greater using the pulsed than using the continuous-wave irradiation. The difference between normal and tumor cells was more evident at 30 micrograms than at 50 micrograms of HPD per ml. The HPD not followed by laser irradiation had no effect on the cell growth rate. The findings of a significant difference in the sensitivity to photoactivated HPD between normal and tumor cells under strictly controlled and highly comparable conditions opens new possibilities to the study of the cellular and molecular mechanisms involved in the phototherapy of tumors. Furthermore, studies in vitro on the active components of the photosensitizer and on their selectivity towards the tumor cells, explained at a cellular level, will lead to better approaches to photochemotherapy in vivo. PMID- 6299539 TI - Action of epidermal growth factor and retinoids on anchorage-dependent and independent growth of nontransformed rat kidney cells. AB - Treatment of two nontransformed rat kidney cell lines with either retinoic acid or epidermal growth factor enhances anchorage-dependent cell growth and reduces cell substratum adhesiveness significantly, whereas the cell morphology is only moderately affected. Simultaneous treatment of these cells with these factors results in a dramatic change in cell morphology and increases cell growth and reduces cell substratum adhesiveness even further. Treatment of nontransformed rat kidney cells with certain retinoids enhances the binding of 125I-labeled mouse epidermal growth factor 2- to 3-fold. Addition of epidermal growth factor stimulates anchorage-independent growth of these cells. Retinoids enhance colony forming ability in soft agar produced by epidermal growth factor, although retinoids by themselves do not affect anchorage-independent growth. Growth in soft agar induced by epidermal growth factor or epidermal growth factor plus retinoic acid treatments appears to be a reversible trait. Addition of epidermal growth factor stimulates secretion of plasminogen activator, whereas this production is not influenced by retinoids. A transformed rat kidney cell line, which exhibits very low epidermal growth factor binding and grows progressively in soft agar, is not significantly affected by retinoid treatment. PMID- 6299540 TI - Detection of a human sarcoma-associated antigen with monoclonal antibodies. AB - Hybridoma cells were derived from a mouse immunized with plasma membranes prepared from the fresh tumor tissues of a patient with malignant fibrous histiocytoma (MFH), a soft tissue sarcoma. Supernatants from the resultant hybridoma clones were screened for positive antibody binding to tumor membranes and negative binding to membrane preparations of normal tissues using a solid phase radioimmunoassay. Two distinct monoclonal IgG1 (kappa) antibodies, 19-14 and 19-24, were identified that showed identical patterns of reactivity with a large panel of tissues. Both antibodies displayed high levels of binding to membranes prepared from a majority of MFH and osteogenic sarcoma tumors tested. Moderate levels of binding were obtained with melanoma, colorectal carcinoma, and first-trimester fetal membranes. Weak or no significant binding was observed with membranes from a variety of autologous and allogeneic normal adult tissues. Antibody reactivities could be specifically removed by absorption with MFH and osteosarcoma membranes but not with adult muscle membranes. An electrophoretic analysis of immunoprecipitated membrane antigens indicated that antibodies 19-14 and 19-24 reacted with the same protein, a monomer with an approximate molecular weight of 102,000. The antigen was detected in membrane preparations of MFH, osteosarcoma, and first trimester fetus, but was not present in normal adult spleen. However, a small amount of antigen of molecular weight 107,000 was precipitated from a normal adult liver preparation, which suggests that related antigens may be present in low levels in some normal tissues. Antibodies 19-14 and 19-24 also specifically bound to intact, cultured MFH cells, indicating that the relevant antigens were expressed on the outer cell surface. PMID- 6299541 TI - Effect of rous sarcoma virus transformation of rat-1 fibroblasts upon their growth factor and anchorage requirements in serum-free medium. AB - The proliferative response of nontransformed rat embryo (Rat-1) cells and avian sarcoma virus-transformed B31 cells to high-density lipoprotein (HDL), transferrin, insulin, epidermal growth factor (EGF), and fibroblast growth factor has been compared. HDL, added in combination with transferrin, supported the active proliferation of low-density cultures of both Rat-1 and B31 cells. No major difference in the sensitivity of Rat-1 or B31 cells to HDL and transferrin was observed when cells were maintained on dishes coated with an extracellular matrix (ECM) obtained from bovine corneal endothelial cells. The two cell types differed in their response to the other known growth-promoting agents, however, in contrast to Rat-1 cells, transformed B31 cells no longer respond to EGF and fibroblast growth factor and respond only inconsistently to the mitogenic stimulus of insulin. Nontransformed Rat-1 cells and transformed B31 cells grown in the presence of medium containing, respectively, HDL, transferrin, insulin, EGF, and dexamethasone or HDL, transferrin, and insulin could be subcultured for more than 50 generations in the complete absence of serum without significant alteration in morphology, growth rate, or tumorigenicity (B31 cells). When plastic or collagen-coated dishes were used as the substrate instead of ECM coated dishes, nontransformed Rat-1 cells grew very slowly in the serum-free medium described above. Dishes coated with collagen were not more efficient than was plastic in supporting growth of Rat-1 cells under these conditions. Coating dishes with fibronectin, however, clearly improved their growth, bringing the final cell density of the cultures up to 50% of that obtained on ECM-coated dishes. In contrast, transformed B31 cells grew significantly in serum-free medium when seeded on plastic or collagen-coated dishes, and the final cell density reached by cells on these substrates was 50% of that of cells maintained on ECM-coated dishes. In addition, B31 cells grew equally well when seeded on fibronectin- or ECM-coated dishes. The transformed cells thus showed less stringent substrate requirements when grown under serum-free conditions than did nontransformed Rat-1 cells. Our data also indicate that HDL, in combination with transferrin, supported efficient anchorage-independent growth of B31 cells. Fibroblast growth factor, but not insulin or EGF, further improved anchorage independent growth of these cells. The capacity of cells to form colonies in semisolid medium when exposed to HDL and transferrin seems to correlate with high tumorigenic potential. PMID- 6299542 TI - Isolation and characterization of a cloned cell line R3327H-G8-A1 derived from the dunning R3327H rat adenocarcinoma. AB - A cloned cell line (R3327H-G8-A1) has been isolated from the Dunning R3327H adenocarcinoma. Light and electron microscopic studies showed that the cell line possessed features common to secretory epithelial cells. These cells, which grow in monolayer culture, produced s.c. hind flank tumors when inoculated inoculated into Copenhagen X Fischer F1 rats.l Chromosomal karyotype analysis confirmed that the cell line is distinctly that of the Rattus norvegicus genus and species. The cells specifically bind testosterone and dexamethasone with equilibrium dissociation constants (Kd) of 0.49 and 0.8 nM, respectively. The numbers of saturable binding sites per cell are 10,000 for testosterone and 60,000 for dexamethasone. The cells also have 5 alpha-reductase activity. These properties are characteristic of the prostate and of the Dunning tumor from which the cells are derived. Cell growth in vitro was stimulated by androgens and inhibited by glucocorticoids at concentrations of 10(-8) M. An int riguing finding was that estradiol and progestins dramatically stimulated growth in the apparent absence of receptors for these hormones. Finally, comparisons between the G8-A1 cells and the tumor induced by the G8-A1 clone and a second generation of cells from this G8-A1-induced tumor showed that the cloned cells retained their properties following passaging in the animal. PMID- 6299543 TI - Epstein-Barr virus-specific leukocyte migration inhibition reaction with tumor biopsy extracts: correlation with the presence of viral DNA. AB - The Epstein-Barr virus (EBV)-specific leukocyte migration inhibition (LMI) reaction was used to detect EBV antigens in human tumor biopsies in parallel with nuclei acid hybridization for EBV DNA. None of six EBV DNA-negative tumors gave any significant LMI reaction. Fourteen of 17 EBV DNA-positive tumors gave a significant difference between the migration of leukocytes from EBV-seropositive versus -seronegative donors. One tumor gave a borderline reaction. The two-LMI negatives in this group had only a marginal EBV DNA content. It is suggested that the EBV-specific LMI test may be useful for detecting EBV genomes in tissue and tumor extracts. PMID- 6299544 TI - Fourth international symposium on nasopharyngeal carcinoma application of field and laboratory studies to the control of NPC. PMID- 6299545 TI - Fiber, steroids, and cancer. AB - Fiber is a generic term for dietary components which are not metabolized by intestinal secretions. Fiber encompasses a variety of substances of unique structure having specific physical characteristics and physiological function. There are a number of epidemiological and case-control studies on diet and colon cancer, many of which suggest that dietary fiber or fiber-rich foods may exert a protective action, but the view is not unanimous. One study finds a negative correlation between cancer mortality and pentose-rich dietary fibers. Animal studies have been carried out using rats of different strain, semipurified or commercial diets, and various carcinogens administered in a variety of ways. Again, there is no unanimity. The results depend upon the type of fiber and the route of carcinogen administration. Some type of fiber bind bile acids and salts, and this property has been investigated as a means of explaining the protective action of fiber. Several recent studies in human subjects suggest that susceptibility to colon cancer is related inversely to the concentration of fecal bile acids rather than to the absolute quantity of fecal bile acids or their structure. Future studies require better standardization of protocols so that intradietary effects can be assessed. PMID- 6299546 TI - Is chest radiotherapy necessary in any or all patients with small cell carcinoma of the lung? Yes. PMID- 6299547 TI - Is thoracic radiation therapy necessary for patients with limited-stage small cell lung cancer? No. PMID- 6299548 TI - Phase II study of vinblastine in small cell carcinoma of the lung. AB - Thirty-five patients with small cell carcinoma of the lung, refractory to previous combination chemotherapy including vincristine, were treated with weekly iv bolus injections of vinblastine. The dose was 4 mg/m2 in the first 22 patients and 7 mg/m2 in the last 13. No responses were observed among 34 evaluable patients. Side effects included moderate leukopenia and minimal neurotoxicity. It is concluded that vinblastine appears to be inferior to other vinca alkaloids in the treatment of small cell carcinoma of the lung. PMID- 6299549 TI - The pharmacologic profile of nabilone: a new antiemetic agent. PMID- 6299550 TI - Pharmacokinetics, metabolism and drug-abuse potential of nabilone. PMID- 6299551 TI - A double-blind, controlled trial of nabilone vs. prochlorperazine for refractory emesis induced by cancer chemotherapy. PMID- 6299552 TI - Treatment of nausea and vomiting caused by cancer chemotherapy. PMID- 6299553 TI - Study design for the evaluation of the efficacy and safety of nabilone. PMID- 6299554 TI - Double-blind, randomized, crossover trial of nabilone vs. placebo in cancer chemotherapy. PMID- 6299555 TI - A multi-institutional Phase III study of nabilone vs. placebo in chemotherapy induced nausea and vomiting. PMID- 6299556 TI - Nabilone vs. placebo in the treatment of chemotherapy-induced nausea and vomiting in cancer patients. PMID- 6299557 TI - Nabilone: an effective antiemetic agent in patients receiving cancer chemotherapy. PMID- 6299558 TI - Use of nabilone in the treatment of chemotherapy-induced vomiting in an outpatient setting. PMID- 6299559 TI - Nutritional and hormonal factors in the prevention of carcinogen-induced neoplastic development in vitro. AB - Cell cultures and in vitro oncogenic transformation offer a powerful tool for exploring conditions and compounds that may inhibit malignant transformation following exposure to environmental or therapeutic agents. We find, using rodent cell cultures, that retinoids inhibit radiation-induced transformation and eradicate the enhancing effect of tumor promotors. These protective effects of the retinoids are not mediated at a chromosomal level but are reflected at the cell membrane level via the membrane-associated transport enzyme Na/K-ATPase. Using these systems, we also find that hypothyroid condition is protective and that thyroid hormone T3 is essential for the induction of both radiation-induced and chemically induced carcinogenesis, and thus may also play a crucial role in cancer induction in vivo. PMID- 6299560 TI - Biochemical markers in colon tumorigenesis: retinoic acid and dihydrotestosterone binding proteins. AB - A specific retinoic acid-binding protein (RABP) and a dihydrotestosterone-binding protein (DHTBP) appeared in colon tumors as well as in tissues surrounding the the tumors, but the proteins were nondetectable in normal adult human colon tissues. We have analyzed a total of 105 human colon tumors and related specimens for the presence of RABP and DHTBP as possible biochemical markers in colon tumorigenesis. The tissue or tumor extracts after incubation with (3H) retinoic acid or (3H) dihydrotestosterone were sedimented on sucrose gradients, and the binding proteins were detected from the 2S (RABP) or 6-7S (DHTBP) radioactive peaks. The overall results of the analysis illustrate that about 78% of the 74 human colon, rectum, cecum, and colorectum tumors analyzed contained RABP in detectable amounts. Thirty percent of the 20 colon tissues isolated from subjects suspected for colon cancer and 18% of the 11 normal colon tissues from autopsies contained detectable amounts of the binding protein. A comparative study on the quantitative amounts of RABP and DHTBP in colon tumors and related tissues indicates that the amount of retinoic acid or dihydrotestosterone bound per mg protein ranged from 0.8 to 5.1 pmoles in the binding protein-positive specimens. However, the relative amounts of the two binding proteins in these tissues were not in the same proportions. Appearance of RABP and/or DHTBP in the surrounding tissues of colon tumors correlated with the amounts of the binding protein(s) in the tumors. RABP of human colon tumor shared the same ligand specificity and other physicochemical characteristics as RABP of other species. PMID- 6299561 TI - The reliability of IgA antibody to Epstein-Barr virus (EBV) capsid antigen as a test for the diagnosis of nasopharyngeal carcinoma (NPC). AB - Since patients with nasopharyngeal carcinoma were first reported to have elevated levels of IgA antibody to Epstein-Barr virus (EBV) in their sera, workers in a number of countries have studied the possibility that this assay could be used in the diagnosis and monitoring of patients with this disease. In the United States, a collaborative project involving seven centers has been established to investigate the potential value of IgA antibody to EBV viral capsid antigen (VCA) as a clinical tool. In this report, we will summarize the results obtained from three studies: a comparison of EBV serology in three laboratories; a retrospective study of 37 nasopharyngeal carcinoma (NPC) patients and controls, and a prospective study of 126 NPC patients and 683 controls, including 149 patients with other malignancies involving the head and neck. The study of testing comparability in three laboratories demonstrated the feasibility of using this assay in a number of laboratories. The retrospective study confirmed the difference in IgA antibody titers between NPC patients and matched controls. The prospective study showed a relationship between IgA antibody titers and histopathology but not disease stage. IgA antibody titers were elevated more frequently in patients with nonkeratinizing or poorly differentiated types of NPC than for the well-differentiated squamous cell carcinomas. While IgA antibodies to EBV VCA appear to be of value in the early detection and diagnosis of NPC, it is possible that additional serologic tests for immunity to EBV, such as IgG antibody to VCA or early antigen (EA), will improve even further the clinical value of EBV serology in the management of NPC. PMID- 6299562 TI - Perspectives and limits of an immunoenzymatic assay (ELISA) for herpes simplex virus (HSV) tumor-associated antigen (TAA). AB - An ELISA has been developed to detect specific antibodies for HSV-TAA in sera of patients with head, neck and urogenital tract carcinomas (Cancer, 45:1980). Further studies have shown that 64/800 controls (8.12%), i.e. healthy people, are positive against 312/425 patients with herpes associated tumors (73.41%). People with herpes recurrens show 18% positivity, precancerous lesions 44% and other cancers 6%. Immunodepression, chemo- and radio-therapy play an inhibitory role on the ELISA positivity for TAA. Radical surgery of the HSV-TAA positive cancers results in lack of specific antibody whereas relapse or metastasis of the tumor yields positive results. The reproducibility of the test is very good and the standard error is low (1.0655). The conclusions allow us to foresee the use of this ELISA for early detection of the HSV-TAA antibodies in certain human carcinomas. PMID- 6299563 TI - Ovarian cysts and masses: diagnosis using fine needle aspiration. AB - The early diagnosis of ovarian tumors is hindered by the inaccessibility of the ovary by noninvasive techniques. The increasing popularity of fine needle biopsy and laparoscopy created a demand for using these techniques to diagnose ovarian masses. Ovarian masses were aspirated prior to, during, or after laparotomy and laparoscopy. The cases included a variety of nonneoplastic cysts and benign and malignant neoplasms. The nuclear and cytoplasmic features of the aspirated cells, the background material and the presence of other structures such as hair, and psammoma bodies and hyaline bodies made it possible to classify most tumors regarding their behaviors as well as their specific type. Malignant neoplasms in general produced more cellular specimens than benign tumors. It was difficult to differentiate between some types of sex cord/stroma neoplasms. Aspiration cytology, which can be performed through a laparoscope transvaginally, transrectally, or transabdominally is a safe, noninvasive, and relatively reliable technique. It should be utilized more in evaluating ovarian masses, when it is desirable to preserve ovarian function in young patients or to minimize surgical trauma to the elderly high-risk patients. PMID- 6299564 TI - Complementary DNA copies of leukemia and sarcoma virus RNA contain sequences of deoxycytidylate and deoxyguanylate. AB - Single-stranded complementary DNA (cDNA) of the RNA of Gazdar murine sarcoma virus, Gz-MSV/MuLV; Moloney murine leukemia virus, M-MuLV; mouse mammary tumor virus, MMTV; and simian sarcoma virus, SSV-1, were synthesized in endogenous reverse transcriptase reaction. Gz-MSV/MuLV cDNA was also synthesized in exogenous in vitro reverse transcriptase reactions. In the endogenous reaction, 30-35S, or 6.0- to 7.9-kilobase-length cDNA transcripts were synthesized in high yield. In comparison, transcripts synthesized in exogenous reactions were 6.7S, or 0.39 kilobases. The complementarity of the transcripts was verified by both RNA/DNA hybridization and protection studies. dG and dC sequences were detected in 50-77% of the cDNA molecules by affinity chromatography, by annealing and masking studies, and by resistance to S1 nuclease. dT and dA sequences were not detected in the transcripts. These findings are discussed in relation to the possible selective blocking of transcription of retrovirus genes without interfering significantly with the transcription of cellular genes. PMID- 6299565 TI - Relationship of benign papillomas to cancer induction in mouse skin. AB - An analysis is presented for evaluating the potency of initiators, promoters, and carcinogens based on the number of tumors that occur as a function of dose and time when compounds are applied topically in an appropriate solvent to mouse skin. A given compound was tested as an initiator (a single dose followed by prolonged promotion), as a whole carcinogen (multiple applications for a prolonged period of time), and as a cocarcinogen (multiple applications for a prolonged period of time in combination with promotion). Compounds tested were 7,12 dimethylbenz(a)anthracene (DMBA), benzo(a)pyrene (B(a)P), 4-nitroquinoline-1 oxide (4-NQO), and betapropriolactone (BPL). The results indicated that various types of papillomas were produced in proportion to the dose applied to DMBA, B(a)P, and BPL. Certain of these papillomas were nonregressible and progressed readily to carcinomas; others regressed and did not progress to cancer. Multiple doses of DMBA and B(a)P produced carcinomas without an antecedent papilloma stage. The latter cancers were produced in proportion to the 2nd or 3rd power of applied carcinogen dose and were accelerated strongly by concomitant action of a promoter. Certain nonregressible papillomas probably represent the first step in a two or three step progression to cancer, although cancers from papillomas occur in proportion to dose. PMID- 6299566 TI - [The significance of vitamin D in tumors]. PMID- 6299567 TI - [Hormonal regulation of energy metabolism in submaximum physical exertion]. PMID- 6299568 TI - [Circulating immunocomplexes in patients after kidney transplantation in relation to infection and rejection]. PMID- 6299569 TI - Extracellular matrix vesicles in rat bone after parathyroidectomy. AB - The enzymatic activity of bone matrix vesicles from parathyroidectomized rats was determined and compared to the activity of vesicles from sham operated and normal animals. The vesicles were isolated from the alveolar bone by collagenase digestion and differential centrifugation and further purified on a discontinuous sucrose density gradient. The amount of extractable protein and the activity of alkaline phosphatase, acid phosphatase, and ATPase in the vesicle fractions thus obtained did not differ significantly from the values characteristic of preparations from control rats. It may therefore be suggested that parathyroid hormone depletion and the associated hypocalcemia have no significant effect on the occurrence and phosphatase activity of bone matrix vesicles. PMID- 6299570 TI - Morphological heterogeneity of the smooth endoplasmic reticulum in the Leydig cells of the boar testis. AB - The fine structure of boar Leydig cells is re-examined in samples post-fixed with OSO4-K4Fe(CN)6, an elective method to preserve the ultrastructural detail of the endocrine testis. The cells show a pleomorphic arrangement of the smooth endoplasmic reticulum. Large "vesicle-like" dilatations are interspersed in the tubular network of this membrane system. The dilatations contain a flocculent material with a dense core and a dispersed peripheral rim. Additional features of boar Leydig cells revealed by OSO4-K4Fe(CN)6 treatment are parallel tubular arrays within residual bodies, and electron-lucent inclusions suggestive of crystals of cholesterol or its esters. These structural characteristics were not identified in previous electron-microscopic studies on Leydig cells of the boar or of other mammals. The unique components of boar interstitial cells described may be the morphological counterpart of the peculiar composition of testicular steroids secreted in this animal, namely low concentrations of androgens and large amounts of pheromones. PMID- 6299572 TI - Recycling receptors: the round-trip itinerary of migrant membrane proteins. PMID- 6299571 TI - Structure of the pars distalis in the adult tammar wallaby (Macropus eugenii). AB - An immunohistochemical, light- and electron-microscopial study was made of the pars distalis in adult tammar wallabies (Macropus eugenii). The pars distalis of this marsupial mammal was divided into three regions, based on the distribution of cell types within the gland. Somatotropic, mammotropic, luteotropic, folliculotropic, corticotropic and thyrotropic cells were identified on the basis of their immunohistochemistry, cytology and ultrastructure. Non-granulated (folliculo-stellate) cells, identified in electron micrographs, were found throughout the pars distalis. Somatotropic cells were predominant in the posterior pars distalis in all animals examined. In the single male specimen and in the non-lactating females examined, small numbers of apparently inactive mammotropic cells were scattered throughout the pars distalis; the same cell type was apparently active and present in considerable numbers in lactating females. Only one morphological type of gonadotropic cell was evident; these cells were scattered throughout the pars distalis, but in largest numbers in the median region. Small numbers of thyrotropic cells were found, most commonly in the anterior pars distalis. Corticotrops were also observed in moderate numbers, predominantly in the anterior regions of the pars distalis. PMID- 6299573 TI - Three regions upstream from the cap site are required for efficient and accurate transcription of the rabbit beta-globin gene in mouse 3T6 cells. AB - Cloned rabbit beta-globin genes, modified in vitro by restructuring or site directed mutagenesis, were introduced into mouse 3T6 cells, and the resulting transcripts were analyzed by nuclease S1 mapping. The first 109 bp preceding the cap site sufficed for maximal beta-globin transcription. This segment contained three functionally important regions of the ATA box region; the CCAAT box region; and the -100 region. The latter consists of an imperfect tandemly repeated sequence of 14 bp and 15 bp, both copies of which are required for optimal promoter function. Each of three regions contains two or more nucleotide positions where the introduction of point mutations reduces transcription by at least a factor of 2. PMID- 6299574 TI - Common and interchangeable elements in the promoters of genes transcribed by RNA polymerase iii. AB - We have shown that the 34 bp internal control region of the somatic 5S RNA gene from Xenopus borealis can be split into two separable components. A maxigene carrying an insertion between nucleotide 71 and nucleotide 74 of the coding region is actively transcribed in the nucleus of X. laevis oocytes, giving rise to a maxitranscript with initiation and termination points identical with those of the wild-type transcript. The first 11 bases of the 5S RNA gene promoter are shown to be structurally and functionally homologous with the first component (box A) of the promoter for tRNA genes. This was shown by constructing hybrid 5S RNA-tRNAPro and tRNAPro-5S RNA genes that were efficiently transcribed in the X. laevis oocytes. Initiation of transcription appears to be a complex phenomenon in which both components of the internal promoter play a role. PMID- 6299575 TI - Induction and modulation of cell-type-specific gene expression in Dictyostelium. AB - We have identified genes that are expressed preferentially in either prestalk or prespore cells in Dictyostelium. The prestalk mRNAs are detectable at 7.5 hr prior to the completion of cell aggregation, while the prespore mRNAs are not detectable until approximately 15 hr of development. Exogenous cAMP in the absence of sustained cell contact is sufficient to induce prestalk-specific gene expression, while multicellularity is required for the induction of prespore specific genes. A gene expressed equally in both cell types, which has the same developmental kinetics as the prestalk genes, is induced in shaking culture in the absence of either cAMP or stable cell associations. Dissociation of aggregates results in the rapid loss of prespore- and prestalk-specific mRNAs, and these can be induced to reaccumulate with the addition of cAMP. We conclude that there are substantial differences in the timing and requirements for tissue specific gene expression in Dictyostelium. PMID- 6299576 TI - A control element within a structural gene: the gal operon of Escherichia coli. AB - The gal operon of Escherichia coli is transcribed from two overlapping promoters, PG1 and PG2. Cyclic AMP and its receptor protein (CRP) modulate the two promoters in opposite directions by binding to a single cat locus. Both the promoters are negatively regulated by a single repressor, the product of the galR gene. An operator site, defined by several mutations, has previously been located upstream from the cat locus. We have isolated and characterized a new set of cis-dominant constitutive mutations of the gal operon and determined their locations by DNA sequencing. From these studies, we propose the existence of a second functional gal operator element at an extraordinary site--within galE, the first structural gene. Both the operators, OE (exterior) and OI (interior), are involved in the repression of PG1 and PG2. This would be the first example of the presence of a functional operator element within a structural protein-coding region. PMID- 6299577 TI - Tn10 transposase acts preferentially on nearby transposon ends in vivo. AB - Transposition of Tn10 requires sites at the termini of the element and one essential transposon-encoded function, "transposase", which acts at those termini. Genetic complementation experiments reveal that this "transposase" function works much more efficiently on transposon ends located near the gene from which they are expressed than on transposon ends located at a distance. This property accounts for the failure of mutant Tn10 elements to be efficiently complemented in trans. The failure of transposase protein to move freely in three dimensions could be explained by one-dimensional diffusion, energy-dependent translocation and/or extreme protein lability. Additional genetic analyses demonstrate that the rate of Tn10 transposition in vivo depends upon the length of the transposon and the amount of transposase protein. Function dependence and length dependence are independent aspects of the transposition process that could correspond to the break/join and replication aspects into which transposition has been separated conceptually. PMID- 6299578 TI - Nucleotide sequence of Rous sarcoma virus. AB - We present the 9312 nucleotide sequence of the Prague C (Pr-C) strain of Rous sarcoma virus (RSV). A comparison of known protein sequences with the nucleotide sequence allows assignment of the coding regions for the gag, pol, env and src genes. The gag gene is terminated by an amber stop codon and is contained within a different reading frame than is the pol gene. The pol and env genes overlap. The sequences surrounding the src gene in the Pr-C and Schmidt-Ruppin (SR-A) strains of RSV have been compared, and they reveal that an element, E, of approximately 153 nucleotides is present on the 3' side of the src gene in Pr-C, and on the 5' side in SR-A. We hypothesize that E was part of a duplicated region of over 250 nucleotides flanking the src gene in an ancestral RSV, and that differential deletion of one copy of E led to its positional difference in Pr-C and SR-A. PMID- 6299579 TI - Frameshift and intragenic suppressor mutations in a Rous sarcoma provirus suggest src encodes two proteins. AB - We have analyzed two mutations in src responsible for phenotypic reversion and subsequent retransformation of a Rous sarcoma virus-transformed rat cell. Comparison of the nucleotide sequences of cloned proviral DNAs reveals a single base pair insertion approximately 438 bp from the 5' end of src in the revertant line 000. This frameshift mutation accounts for an 18 kd protein as the prematurely terminated product of src. The mutation is suppressed in the retransformed line (000*) by a 242 bp duplication that corrects the reading frame to permit synthesis of a 68 kd src protein. A 43 kd src protein with a normal carboxyl terminus is also present in 000 and 000* cells. To make this protein, translation must begin at an internal AUG, found just upstream from the frameshift mutation; in both lines, two src proteins appear to be initiated independently from the same mRNA species. Our results imply that a protein of 7 kd is synthesized from a second reading frame within wild-type src. PMID- 6299580 TI - Structure and sequence of the cellular gene homologous to the RSV src gene and the mechanism for generating the transforming virus. AB - We determined the nucleotide sequences of all coding regions and a significant part of the flanking regions of the chicken c-src gene, which is a cellular homolog of the v-src gene of Rous sarcoma virus. The c-src gene consists of 12 exons; the boundaries of the exons were determined by assuming that the amino acid sequence of its product, pp60c-src, is basically the same as that of pp60v src. The deduced amino acid sequence of pp60c-src was very similar to that of pp60v-src, but the last 19 carboxy-terminal amino acids of pp60c-src were replaced by a new set of 12 amino acids of pp60v-src. The sequence encoding the carboxy-terminal sequence of pp60v-src was found 900 bp downstream from the termination codon of the c-src gene. We suggest that the c-src sequence was captured by a virus through recombination at both sides of the c-src gene, and that the recombinations occurred at the level of proviral DNA. PMID- 6299581 TI - Differentiation of cloned populations of immature B cells after transformation with Abelson murine leukemia virus. AB - The nature of the target cell for Abelson virus transformation and the effect of transformation on B cell differentiation were studied with six cloned lines of nontransformed immature B lymphocytes. Three clones were at the pre-B cell stage of maturation prior to A-MuLV infection; two were at the B cell stage, and one appeared to represent a stage prior to rearrangement of the mu heavy chain gene. All six cloned lines could be transformed by Abelson virus. Many of the transformants of the pre-B cell clones underwent kappa light chain gene rearrangement and expression following viral infection. Distinct light chain gene rearrangements were segregated by further subcloning these transformed lines. Abelson virus infection of one cloned cell line believed to represent a stage of maturation prior to the pre-B cell stage produced pre-B cell transformants with a variety of heavy chain gene rearrangements. Thus B lymphoid target cells for Abelson virus are not restricted to a single developmental stage, and some transformed subclones can undergo extensive immunoglobulin gene rearrangements shortly after viral infection. PMID- 6299582 TI - The LDL receptor locus in familial hypercholesterolemia: multiple mutations disrupt transport and processing of a membrane receptor. AB - The receptor for low-density lipoprotein (LDL) is synthesized as a 120 kd precursor that undergoes a 40 kd posttranslational increase in apparent molecular weight en route to the cell surface. We describe seven mutations that disrupt synthesis, processing and transport of the receptor in fibroblasts from 77 subjects with the clinical diagnosis of homozygous familial hypercholesterolemia. One mutation obliterates synthesis of immunoprecipitable precursor. Three mutations specify precursors (100, 120 and 135 kd) that fail to undergo normal processing and fail to reach the cell surface. The other three mutations specify precursors (100, 120, and 170 kd) that undergo a normal 40 kd increase in molecular weight and reach the surface, but do not bind LDL normally. Pedigree studies show that each mutation segregates as an allele at the LDL receptor locus. These data imply that signals for transport of receptors from endoplasmic reticulum to the cell surface are contained within the amino acid sequences of the receptors, and that mutations affecting these sequences can disrupt receptor transport. PMID- 6299583 TI - Differences between liver gap junction protein and lens MIP 26 from rat: implications for tissue specificity of gap junctions. AB - Liver gap junctions and gap-junction-like structures from eye lenses are each comprised of a single major protein (Mr 28,000 and 26,000, respectively). These proteins display different two-dimensional peptide fingerprints, distinct amino acid compositions, nonhomologous N-terminal amino acid sequences and different sensitivities to proteases when part of the intact junction. However, the junctional protein of each tissue is well conserved between species, as demonstrated previously for lens and now for liver in several mammalian species. The possiblity of tissue-specific gap junction proteins is discussed in the light of data suggesting that rat heart gap junctions are comprised of yet a third protein. PMID- 6299584 TI - O-linked oligosaccharides are acquired by herpes simplex virus glycoproteins in the Golgi apparatus. AB - The O-linked oligosaccharides on mature forms of herpes simplex virus type 1 (HSV1) glycoproteins were characterized, and were found to account largely for the lower electrophoretic mobilities of these forms relative to the mobilities of immature forms. Other posttranslational modifications of HSV1 glycoproteins (designated gB, gC, gD and gE) were related temporally to the discrete shifts in electrophoretic mobilities that signal acquisition of the O-linked oligosaccharides. Fatty acid acylation (principally of gE) could be detected just prior to the shifts, whereas conversion of high-mannose-type N-linked oligosaccharides to the complex type occurred coincident with the shifts. The addition of O-linked oligosaccharides did not occur in cells treated with the ionophore monensin or in a ricin-resistant cell line defective in the processing of N-linked oligosaccharides. We conclude that extension of O-linked oligosaccharide chains on HSV1 glycoproteins, and probably also attachment of the first O-linked sugars, occurs as a late posttranslational modification in the Golgi apparatus. PMID- 6299585 TI - Anchorage-independent colony formation of SV40 transformed BALB/c-3T3 cells in serum-free medium: role of cell- and serum-derived factors. AB - The colony-forming response of SV40 transformed BALB/c-3T3 cells in agarose suspension culture was studied in a serum-free medium (with insulin, transferrin and serum albumin as the only macromolecular supplements) that was optimized for colony formation of fibronectin-attached monolayer cultures. In this serum-free medium, the SV3T3 cells fail to form colonies in agarose suspension. However, they can be induced to anchorage-independent colony formation by the growth factors that are additionally required by their untransformed counterparts for proliferation in monolayer culture. The SV3T3 cells are also rendered anchorage independent for colony formation in serum-free medium by conditioned medium from dense monolayer serum-free SV3T3 cultures. These experiments suggest that it is the cell-substrate interaction that is responsible for the growth factor autonomy of fibronectin-attached transformed cells. PMID- 6299586 TI - Kinetics and concentration effects of TPA-induced differentiation of cultured human neuroblastoma cells. AB - SH-SY5Y human neuroblastoma cells differentiate morphologically and biochemically in the presence of 12-0-tetradecanoylphorbol-13-acetate (TPA). The degree of differentiation, as demonstrated by the appearance of cell surface projections, growth inhibition and increase in noradrenalin concentration, was dependent on the TPA concentration and had an optimum at 1.6 X 10(-8) M of TPA. At that concentration neuron specific enolase (NSE) increased to a maximum level after 10 days of culture with no further changes in the NSE level during additional culture for 10 days. In contrast, the noradrenalin concentration reached a maximum after 4 days of TPA treatment and decreased during longer exposures to TPA. Based on the facts that the phorbolester induced differentiation shows stereo specificity and was optimal at the same concentration range as common polypeptide hormones, a putative TPA-hormone receptor interaction is discussed. An opposite effect of TPA on the SH-SY5Y cells, antagonizing the differentiation effect, is further suggested to explain the decrease in differentiation observed at TPA concentrations higher than 1.6 X 10(-8) M. PMID- 6299587 TI - Possible role of microtubules in tumor cell surface membrane shedding, permeability, and lympholysis. AB - Low temperature (0 degrees C/1 hr) induced changes in the mastocytoma P815 cell surface morphology which mimicked those induced by alloimmune T-lymphocytes during the cytolytic process. These morphological changes consist of the formation and shedding of membrane vesicles (MV) from the P815 cell surface. When the low-temperature-induced MV shedding process takes place at 22 degrees C, it occurs without changes in membrane permeability to 51Cr, whereas at 37 degrees C, the kinetics of 51Cr release resemble those induced by alloimmune T lymphocytes. The effects of two microtubule-stabilizing agents, deuterium oxide and hexylene glycol, were investigated on the low-temperature-induced MV shedding process as well as on the lymphocyte-mediated cytolysis (LMC) of target cells. These agents were found to inhibit (a) the low-temperature-induced MV shedding, (b) the low temperature-induced membrane permeability changes to 51Cr, and (c) the LMC process. These data indicate that disruption of microtubules may be involved in all these events. PMID- 6299588 TI - Transient decrease of beta-adrenergic receptors induced by isoproterenol is not energy dependent. PMID- 6299589 TI - Multicellular tumour spheroids derived from human brain tumours. AB - Multicellular tumour spheroids were prepared from a total of 46 human brain tumour biopsies by collagenase digestion and plating into agar coated flasks. Both primary malignant and secondary tumours formed spheroids with some correlation between the malignancy of tumour and the ability to undergo spheroid formation. The spheroids were capable of progressive growth, the rate of which was dependent, to some extent, on environmental conditions and was reflected by an increase in cell number within the spheroids. Spheroids prepared in this way may prove to be useful models for in vitro chemosensitivity and the general biology of brain tumours. PMID- 6299590 TI - Mitotic apparatus and nucleoli compartmentalization of 50,000-dalton type II regulatory subunit of cAMP-dependent protein kinase in estrogen receptor negative MDA-MB-231 human breast cancer cells. AB - Affinity purified RI and RII antibodies of regulatory subunits (R) of type I (RI) and type II (RII) cAMP-dependent protein kinase were utilized to determine the immunological characterization and specific compartmentalization of R in estrogen receptor negative MDA-MB-231 human breast cancer cells. The 8-azido-(32P)-cAMP binding analysis of MDA-MB-231 cell extracts exhibited 47,000- and 50,000-dalton cAMP receptor proteins. RI and RII antibodies, by immunoprecipitation, detected the 47,000- and 50,000-dalton proteins, respectively. The 47,000-dalton protein was identified as RI as it showed a similar molecular weight as of bovine RI on SDS-polyacrylamide gel electrophoresis. Although 50,000-dalton protein did not co migrate with bovine heart 54,000-dalton RII, it was identified as RII of MDA-MB 231 cells since it was specifically precipitated with RII antibody but not with RI antibody. An indirect immunofluorescence revealed that during different phases of growth of MDA-MB-231 cells, 50,000-dalton RII was specifically compartmentalized in the mitotic spindle and nucleoli of the cells whereas RI did not exhibit a specific compartmentalization in the cells, but was distributed throughout the cell components. These results suggest specific role(s) of 50,000 dalton RII at the nuclei of MDA-MB-231 cells. PMID- 6299591 TI - Establishment of a human cell line after transformation by a plasmid containing the early region of the SV40 genome. AB - A human clonal cell line, designated TAH9, has been established from cells of adipose tissue by fusion with E. coli protoplasts containing a recombinant plasmid carrying the early genes of Simian virus 40. The establishment of the cell line is preceeded by a delayed growth crisis. The results indicate that, after cell transformation, another event(s) of low frequency is required for cell immortalization. Some biological properties of TAH9 cells are also described. PMID- 6299592 TI - Synthesis of the docosapeptide corresponding to the amino acid sequence of salmon corticotropin-like intermediate lobe peptide (CLIP). PMID- 6299593 TI - [Effect of thyroid hormone (T3) on brain cyclic nucleotide system and its relation to catecholamine supersensitivity]. PMID- 6299594 TI - [Effect of huangqi on plasma and tissue cAMP and cGMP in mice]. PMID- 6299595 TI - [Studies on serum complement fixation antibodies to Epstein-Barr virus in tupaias]. PMID- 6299596 TI - [Candida albicans septicemia with bilateral chorioretinal involvement treated with amphotericin B and ketoconazole]. PMID- 6299597 TI - [Treatment of corneal ulcer with a collagenase inhibitor: cysteine hydrochloride]. PMID- 6299598 TI - Therapeutic efficacy and pharmacokinetics of vindesine and vindesine-cisplatin in previously treated patients with non-small cell lung carcinoma. AB - Twenty-nine patients with non-small cell lung cancer refractory to prior therapy were treated with either vindesine (VDS) alone (3 mg/m2 every week) or the combination of VDS plus cisplatin (DDP) (100 mg/m2 every 28 days). Serial blood and urine samples were collected to assess the pharmacokinetics of VDS and DDP. All patients were evaluable for toxicity and 27 were evaluable for response. No objective antitumor responses were observed. Peripheral neuropathy manifested by paresthesias, muscle weakness, and constipation were observed in 20 treated patients, and hematologic toxicity consisting of thrombocytopenia and/or leukopenia occurred in 18 patients. The plasma and urinary pharmacokinetics of VDS and DDP measured in this study indicate that VDS and DDP do not interfere with each other and that the pharmacokinetics in previously treated and untreated patients are similar. The antitumor responses and degree of toxicity observed in this trial compare unfavorably with previously reported VDS and VDS-DDP trials in previously untreated patients with this disease and suggest that prior exposure to chemotherapy might both decrease antitumor activity and enhance toxicity of these chemotherapeutic agents. PMID- 6299599 TI - Effect of vitamin A palmitate on mutagenesis induced by polycyclic aromatic hydrocarbons in human cells. AB - The effect of vitamin A palmitate (VAP) on mutagenesis induced by polycyclic aromatic hydrocarbons (PAH) was examined in a human epithelial-like cell line (EUE). Cultures were exposed to benzo[a]pyrene (BP), 3-methylcholanthrene (MCA), 7,12-dimethylbenz[a]anthracene, with or without VAP, and mutation frequencies were determined by selection against diphtheria toxin. A strong inhibition of mutagenesis was observed particularly with MCA and BP. VAP, in the same cell line, reduced 3H-labeled PAH binding to DNA. PMID- 6299600 TI - In vitro DNA reaction with an ultimate carcinogen model of 4-nitroquinoline-1 oxide: the 4-acetoxyaminoquinoline-1-oxide. Enzymatic degradation of the modified DNA. AB - 2-3H-Labelled 4-acetoxyaminoquinoline-1-oxide (Ac-4 HAQO), the ultimate carcinogen model of 4-nitroquinoline-1-oxide, was reacted in vitro with native and denatured DNA. We found that Ac-4 HAQO is 2- to 3-fold more reactive than diAc-4 HAQO, another ultimate carcinogen model of 4 NQO which was previously studied [Galiegue et al. (1980) Biochim. Biophys. Acta, 609, 383-391]. Ac-4 HAQO modified DNA is thermally destabilized: when 1% of the bases of DNA were modified by Ac-4 HAQO, its melting temperature decreased 1.2 degrees C. Enzymatic degradation of Ac-4 HAQO-modified native and denatured DNA's to nucleosides was performed. The hydrolysates were analyzed, first with a simple chromatographic system, and then by h.p.l.c. The compounds recovered from the modified polymers were characterized by h.p.l.c. and a variation in their respective amounts as a function of the secondary structure of DNA was observed. Especially, the N (deoxyguanosin-(C8-yl)-4-aminoquinoline-1-oxide, the so called dG III adduct, was recovered from DNA, and its amount was evaluated to be approximately 3.5-fold greater in the case of denatured DNA than in the case of native DNA. PMID- 6299601 TI - Prostaglandin H synthase metabolism of the urinary bladder carcinogens benzidine and ANFT. AB - Prostaglandin H synthase (PHS) and horseradish peroxidase catalyze the oxidation of benzidine to the same free radical species. No radical was observed if either benzidine, H2O2 or enzyme was omitted. The similarity of the fine structure of this radical to a computer-simulated model suggests the presence of a free cation radical of benzidine. Neither superoxide nor hydroxyl radicals appear to be involved in the co-oxidation of benzidine or 2-amino-4-(5-nitro-2-furyl)-thiazole (ANFT) by PHS. Production of the benzidine radical by PHS was inhibited by ANFT, acetaminophen, cyanide and ascorbate. ANFT was metabolized by PHS but not by horseradish peroxidase. ANFT had no effect on either radical production or 14C metabolism of benzidine by horseradish peroxidase. These results indicate that different peroxidases may exhibit specificity with respect to the carcinogens they activate. The free radical cation of benzidine may be the electrophilic intermediate responsible for PHS-catalyzed binding of benzidine to protein and nucleic acids. PMID- 6299603 TI - The paradox of steroid therapy: inhibition of oxygen free radicals. PMID- 6299602 TI - A comparison between the activation of benzo[a]pyrene in organ cultures and microsomes from the tracheal epithelium of rats and hamsters. AB - Epidermoid cancers of tracheal origin produced in Syrian golden hamsters and Fischer strain 344 rats are models for human bronchogenic carcinomas. These two species differ, however, in the sensitivity of their tracheal epithelia to tumor induction elicited by intratracheal benzo[a]pyrene (BP)-ferric oxide administration. The tracheas of hamsters are quite sensitive to the carcinogenic effects of BP-ferric oxide, but rat tracheas are apparently resistant to effects of comparable treatments by this route of administration. Rat tracheas are not completely resistant to polynuclear hydrocarbon carcinogenesis because in the heterotopic tracheal graft model, epidermoid carcinomas have been produced frequently. To determine whether differences in BP metabolism could explain this difference between species, quantitative kinetic and chromatographic studies of benzo[a]pyrene monoxygenase activity were carried out in epithelial microsomes and cells from organ cultures of rat and hamster tracheas. The Vmax was 2-fold greater in hamster tracheal cells than in rat tracheal cells whereas the Km values were identical. H.p.l.c. profiles from microsomes of rat and hamster tracheal epithelial cells incubated with BP exhibited extreme differences. Hamster tracheal microsomes produced large proportions of BP-quinones, BP phenols, and BP-diols but rat tracheal microsomes produced mostly 3-OH BP. The total metabolic rate for BP in rat tracheal organ cultures was half that in cultures of hamster tracheas. The metabolites isolated in organ cultures of hamster and rat tracheas well reflected secondary reactions of conjugation and recycling. When evaluated with respect to the amount of tissue used, the most striking difference between rat and hamster tracheal organ cultures was in the amount of products which co-chromatographed with bay-region BP-tetrols. The amount of BP-tetrols produced by hamster tracheas was 0.22 pmol/mg tissue/24 h, and by rat, 0.012 pmol/mg tissue/24 h. The level of BP-DNA binding (in pmol/microgram DNA/24 h) catalyzed by hamster tracheal cells was 26.6 +/- 11.4, and by rat tracheas was 1.55 +/- 1.29. These interspecies differences associated with the formation of BP-diol epoxide, a presumed ultimate carcinogenic form of BP, are consistent with the differences between these two rodent species in susceptibility to carcinogenesis in this tissue. PMID- 6299604 TI - Diltiazem selectively inhibits cerebrovascular extrinsic but not intrinsic myogenic tone. A review. AB - The tone of blood vessels resulting from extrinsic influences is dependent on activator calcium derived from sequestered cellular and also extracellular sources. Experiments on rabbit blood vessels in vitro show that--in comparison to systemic vessels--the contractions of cerebral arteries are proportionately more dependent on extracellular calcium and have sequestered calcium sources that are more easily depleted. Both these observations suggest that activator calcium mechanisms differ at least quantitatively in cerebral compared to noncerebral vessels. Diltiazem antagonizes agonist-induced tone in cerebral arteries at a lower concentration than is required in systemic arteries and veins. For example, the concentration of diltiazem to halve the response of the rabbit basilar artery to norepinephrine (ED50) is 1 X 10(-8) M, whereas those for the ear and mesenteric arteries and the saphenous vein are 10.4, 2.0, and 1.8 X 10(-7) M, respectively. A similar selectivity has been found to K+ and serotonin-induced contractions. It has been argued that the neuroeffector mechanism in the portal vein reflects that in systemic resistance vessels. The drug is effective against the spontaneous rhythmic activity of the longitudinal muscle of the vein, reducing it to half at 1.5 X 10(-7) M. Diltiazem is relatively ineffective in antagonizing intrinsic smooth muscle tone, both cerebral (basilar) and extracerebral (facial vein) requiring ED50 values of 4 and 5 X 10(-4) M, respectively. In summary, our in vitro experiments on rabbit tissue show that diltiazem selectively antagonizes cerebrovascular extrinsic but not intrinsic tone. PMID- 6299605 TI - Calcium antagonists. Some chemical-pharmacologic aspects. AB - Ca++ serves multiple roles as a regulator and initiator of cellular events. A variety of mechanisms serve to control cellular Ca++ levels and there exists a corresponding diversity of drugs which possess, with varying degrees of potency and selectivity, Ca++-antagonistic properties. Particular interest attaches to a group of agents designated as Ca++-channel or Ca++-entry blockers and includes verapamil, nifedipine, diltiazem, cinnarizine, and prenylamine. These agents function by blockade of the potential dependent Ca++-channel. However, their obvious chemical heterogeneity suggests that several sites and mechanisms of action may exist. A review of some basic questions concerning the action of the Ca++ channel antagonists is presented. The existence of discrete structure activity relationships is consistent with the concept that these agents have specific sites of action, rather than serving, for example, as, nonspecific membrane-stabilizing agents. This view is further supported by their selectivity of action seen both in tissue selectivity and selectivity of antagonism of agonist responses. Studies of the relationship of these compounds to Ca++ show that they appear to function competitively against Ca++ and to block cellular Ca++ uptake. Moreover, both contractile responses and cellular Ca++ uptake appear equisensitive to these antagonists, providing further evidence that inhibition of Ca++ uptake underlies their inhibition of mechanical response. Of particular importance is the question of selectivity of action. It is quite clear that the Ca++ channel antagonists do not show equal activity toward all Ca++ channels and that major differences in selectivity occur between cardiac and smooth muscle. It is possible that selectivity of antagonism may occur between different vascular beds. Furthermore, Ca++ channels controlling stimulus-secretion coupling appear to be less sensitive to these antagonists than the channels involved in excitation-contraction coupling. Finally, the actions of Ca++ antagonists on non Ca++-mediated processes, including Na+-dependent and receptor-binding events, are noted. These actions are seen at higher concentrations than those needed to inhibit Ca++ channel events, and their contribution to the therapeutic actions exerted by these antagonists is unknown. PMID- 6299606 TI - Interval- and voltage-dependent effects of the calcium channel-blocking agents D600 and AQA 39 on mammalian ventricular muscle. AB - Myocardial depressant actions of D600 and AQA 39, a D600-like compound, have been studied in rabbit and cat ventricular muscle using simultaneous action potential and tension measurements and the single hybrid sucrose gap technique to measure the slow inward current (Isi). D600-induced depression of Isi is shown to be greater after a depolarization (action potential or clamp pulse) than before. When a muscle is stimulated with depolarizing clamp steps, the degree of Isi depression increases with the number and voltage of the activating pulses. The degree of si-channel block also depends on the length of the diastolic interval and on the membrane potential during diastole. At diastolic potentials between 50 and -90 mV, there is a restoration of channels to the conducting pool. The rate of this dissociation of drug from si-channels is faster at more negative potentials (T1/2 of 15 sec at -90 mV, 350 sec at -50 mV). AQA 39 has a chemical structure similar to that of D600 but is much less hydrophobic. It also blocks si channels in a frequency- and voltage-dependent manner. The major difference between the two compounds is in the voltage-dependent rate of recovery from si channel block. In comparison to the rate of unblock of D600-bound channels, unblock with AQA 39 is about twice as fast at -50 mV and nearly 100 times as fast at -80 mV. PMID- 6299607 TI - Functional interaction between calcium antagonists and the vasoconstriction induced by the stimulation of postsynaptic alpha 2-adrenoceptors. PMID- 6299608 TI - Actions of diltiazem on single smooth muscle cells and on neuromuscular transmission in the vascular bed. PMID- 6299609 TI - Active transport and inotropic state in guinea pig left atrium. AB - Although the positive inotropic effect of cardiac glycosides correlates well with inhibition of Na+ pump activity in many preparations, digitalis at low concentrations (10(-9) to 10(-8) M) may produce an apparent stimulation of monovalent cation transport in isolated intact myocardium or produce an inotropic effect that does not correlate with pump inhibition. Digitalis is known to modify tissue metabolism of endogenous neurotransmitters that may affect inotropic state, Na,K-ATPase activity, and K+ permeability. We examined the interactions of low concentrations of ouabain with adrenergic and cholinergic influences in isolated guinea pig left atria stimulated at 3.3 Hz in which inotropic state and monovalent cation transport (measured as 86Rb+ uptake) were assessed simultaneously. Ouabain (10(-9) M) stimulated Rb+ transport (+25%) without an inotropic response; the stimulatory effect on transport was abolished by propranolol or atropine pretreatment. In atria pretreated with atropine, 10(-8) M ouabain produced a small positive inotropic effect (+10%) without measurable associated Na+-K+ pump inhibition. This inotropic response was abolished in catecholamine-depleted atria. Ouabain (10(-7) M) always produced a positive inotropic response (about +25%) independent of catecholamine depletion, beta adrenergic blockade, or muscarinic blockade, but Rb+ uptake inhibition was observed only in beta-adrenergically-blocked atria. In all preparations, ouabain concentrations greater than 10(-7) M caused an inotropic response associated with pump inhibition. At concentrations 3 X 10(-7) M and higher, mechanical toxicity was observed in all preparations except those pretreated with propranolol. Incubation with low concentrations of ouabain did not modify the inotropic response to isoproterenol. At concentrations of isoproterenol sufficient to stimulate Rb+ transport by 25%, there was a large (+80%) inotropic response. We conclude first, that, in guinea pig atria exposed to ouabain, the mechanism as well as the extent of inotropic response and of monovalent cation transport modification is concentration dependent, second, that at low concentrations (1-10 X 10(-9) M), in vitro inotropic and monovalent cation transport responses are in part mediated by an effect of ouabain on endogenous neurotransmitters; and third, that in this preparation at concentrations between 10(-9) and 10(-7) M ouabain, monovalent cation transport as measured by tissue 86Rb+ uptake does not correlate with inotropic response. PMID- 6299610 TI - Coronary artery spasm in intact dogs induced by potassium and serotonin. AB - Although coronary artery spasm has been implicated as an important cause of myocardial ischemia in humans, an animal model of reversible segmental coronary constriction has not been described. To provoke coronary spasm in open-chest dogs, selected vasoconstricting agents adsorbed to viscous ion exchange gels were applied topically to the surface of epicardial coronary arteries. The procedure provided a sustained localized release of drug, and minimized effects on contiguous myocardium or on the systemic circulation. Segmental arterial constrictor responses were evaluated by sonomicrometry, arteriography, and electromagnetic flow measurements. Potassium evoked sustained constrictions or spasms, and concomitantly reduced flow by -42 +/- 4% (SE; n = 34). Serotonin likewise produced sustained decreases in flow of -22 +/- 6% (SE; n = 5). Other constrictors, including norepinephrine and angiotensin, failed to evoke sustained constrictions. Spasms nearly abolished reactive hyperemic responses elicited by temporary complete occlusion of the artery. Intravenous nitroglycerin and dihydropyridine calcium antagonists promptly relieved the spasms. Scanning electronmicroscopic examination of the intimal surface of arteries undergoing sustained spasm revealed no platelet thrombi. Thus, nonthrombotic, vasodilator sensitive segmental coronary spasms were elicited by endogenous constrictors which may play a role in regulating flow to ischemic myocardium. PMID- 6299611 TI - Facial vein of the rabbit. Intracellularly recorded hyperpolarization of smooth muscle cells induced by beta-adrenergic receptor stimulation. AB - A segment of the facial vein of the rabbit gains myogenic tone when placed under tension in a tissue bath (Winquist and Bevan, 1977); low frequency electrical stimulation of intramural nerves, or application of norepinephrine or isoproterenol produces relaxation, which is blocked by propranolol (1.0 microM). We measured membrane potential of single smooth muscle cells with microelectrodes during this beta-adrenergic receptor-mediated relaxation of the vein segment. Hyperpolarization (up to 30 mV) invariably accompanied relaxation. Average maximum resting potential recorded in normal Krebs solution was -46.8 +/- 3.6 mV (mean +/- SE); this increased to -70.8 +/- 3.0 mV when high concentrations (greater than 10 nM) of isoproterenol were added to the bath. Tone and membrane potential were very sensitive to intramural nerve stimulation. At 3-4 Hz, relaxation was about half maximal and hyperpolarization was 6-9 mV. The maximum hyperpolarization to nerve stimulation was 20-30 mV, about the same magnitude as that produced by isoproterenol. Hyperpolarization clearly preceded relaxation by about 0.2 second. The magnitude of hyperpolarization in facial vein is much larger (2-10 times or more) than that recorded in other tissues during inhibition produced by stimulation of beta-adrenergic receptors. The large hyperpolarization may augment the effectiveness of inhibition (by curtailing potential dependent influx of Ca++o, for example) in this tissue compared to that in other tissues in which hyperpolarization is slight or absent. The ionic mechanism underlying hyperpolarization resulting from stimulation of beta-adrenergic receptors remains unclear. PMID- 6299612 TI - Differentiation of hemodynamic, humoral and metabolic responses to beta 1- and beta 2-adrenergic stimulation in man using atenolol and propranolol. AB - The respective contributions of beta-adrenoceptor subtypes to the hemodynamic, humoral and metabolic consequences of adrenergic stimulation during graded exercise in man were investigated using nonselective beta-adrenoceptor blockade with propranolol and beta 1-adrenoceptor blockade with atenolol. Doses of these agents that produced comparable suppression of beta 1 response as measured by antagonism of cardioacceleration during exercise were selected. Six healthy, nonsmoking males received these drugs in a placebo-controlled, Latin-square, randomized manner using a double-blind protocol. Both drugs produced comparable reductions of systolic blood pressure and elevation of diastolic blood pressure compared with placebo as exercise load increased. Propranolol produced higher peak epinephrine levels than atenolol or placebo (808 +/- 162, 640 +/- 190 and 584 +/- 153 pg/ml, respectively, p = 0.03), but norepinephrine levels did not show significant differences. Plasma renin activity was similarly suppressed both at rest and during all grades of exercise by both drugs. Lactate levels during moderate exercise were significantly lower after propranolol than after either atenolol or placebo (p = 0.03), but were similar at heavy work loads. Plasma glucose values rose on placebo (from 96.5 +/- 2.1 to 97.7 +/- 2.7 mg/dl) and on atenolol (from 99.7 +/- 2.2 to 102.1 +/- 4.8 mg/dl), but fell on propranolol (from 96.4 +/- 1.9 mg/dl to 87.2 +/- 2.5 mg/dl, p less than 0.01). These results indicate that blockade of vascular smooth muscle beta 2 receptors does not substantially alter hemodynamics during intense short-term exercise. Stimulation of renin release and lipolysis are produced through beta 1-adrenoceptor mechanisms, whereas beta 2 adrenoceptors are important in the provision of carbohydrate as an energy substrate for exercising muscle. PMID- 6299614 TI - Radioallergosorbent testing with p-tolyl monoisocyanate in toluene diisocyanate workers. AB - Sera from 149 occupationally exposed toluene diisocyanate (TDI) workers were tested by RAST with p-tolyl monoisocyanate human serum albumin (TMI-HSA). Of the seven positives, one had a history suggestive of TDI asthma and one had a rash which may have been TDI related. These results indicate that false positive RAST with TMI-HSA may occur in asymptomatic workers. PMID- 6299613 TI - Exercise testing in asymptomatic or minimally symptomatic aortic regurgitation: relationship of left ventricular ejection fraction to left ventricular filling pressure during exercise. AB - Exercise radionuclide angiography is being used to evaluate left ventricular function in patients with aortic regurgitation. Ejection fraction is the most common variable analyzed. To better understand the rest and exercise ejection fraction in this setting, 20 patients with asymptomatic or minimally symptomatic severe aortic regurgitation were studied. All underwent simultaneous supine exercise radionuclide angiography and pulmonary gas exchange measurement and underwent rest and exercise measurement of pulmonary artery wedge pressure (PAWP) during cardiac catheterization. Eight patients had a peak exercise PAWP less than 15 mm Hg (group 1) and 12 had a peak exercise PAWP greater than or equal to 15 mm Hg (group 2). Group 1 patients were younger and more were in New York Heart Association class I. Group 1 patients also had a higher mean rest ejection fraction (0.64 +/- 0.08 vs 0.49 +/- 0.13, p less than 0.01, higher exercise ejection fraction (0.63 +/- 0.10 vs 0.40 +/- 0.18, p less than 0.01), lower end systolic volume (38 +/- 13 vs 79 +/- 36 ml/m2, p less than 0.01) and higher peak oxygen uptake (24.9 +/- 5.1 vs 16.6 +/- 4.9 ml/kg/min, p less than 0.01) than group 2 patients. However, the two groups had similar cardiothoracic ratios, changes in ejection fractions with exercise, and rest and exercise regurgitant indexes. Using multiple regression analysis, the best correlate of the exercise PAWP was peak oxygen uptake (r = -0.78, p less than 0.01). No other measurement added significantly to the regression. When peak oxygen uptake was excluded, rest and exercise ejection fraction also correlated significantly (r = -0.62 and r = 0.60, respectively, p less than 0.01). Patients with asymptomatic or minimally symptomatic severe aortic regurgitation have a wide spectrum of cardiac performance in terms of the PAWP during exercise. The absolute rest and exercise ejection fraction and the level of exercise achieved are noninvasive variables that correlate with exercise PAWP in aortic regurgitation, but the change in ejection fraction with exercise by itself is not. PMID- 6299615 TI - Absence of IgG antibodies to TDI-HSA in a radioimmunological study. AB - Sera of ninety-one workers with respiratory symptoms related to occupational exposure to toluene diisocyanate (TDI) were tested by a solid phase radioimmunoassay using polyvinyl plates, in an attempt to demonstrate specific IgG antibodies to a conjugate of TDI with human serum albumin (TDI-HSA). Different conjugates were prepared. In our radioimmunoassay a TDI-HSA 1600 (1600 micrograms TDI/mg HSA), obtained at alkaline pH with borate buffer, was used; this conjugate was immunologically reactive and did not contain protein macro aggregates. No difference in percent of binding of radiolabelled anti-human IgG to TDI-HSA coated on plastic plates was found between exposed workers (asthmatic and non-asthmatic) and fifteen normal subjects. This may suggest that either TDI induced bronchial asthma is not mediated by IgG antibodies, or TDI-HSA conjugate is unsuitable to be detected by anti-isocyanate IgG. PMID- 6299616 TI - Deficiency of glycerol kinase (EC 2.7.1.30). AB - We describe the case of a 10-year-old boy who had been admitted on several occasions with a diagnosis of gastroenteritis. He had been severely ill, and on one occasion lost consciousness. He had a metabolic acidosis on these occasions. Examination of the urine by gas chromatography-mass spectrometry showed a large peak, identified as glycerol. The concentration of glycerol in the urine was 40 280 mmol/L and the concentration in plasma about 2 mmol/L. He was subjected to a fast of 21 h, at the end of which he expressed feelings of discomfort and nausea, began vomiting, and became somnolent. During this period the blood glucose concentration was only slightly decreased, the plasma glycerol concentration increased to 4.9 mmol/L, and the plasma lactate concentration increased to 3.8 mmol/L. During work on a bicycle ergometer for 35 min (40 W) he complained of muscle pain and became nauseated, but there was no significant increase in the concentration of plasma glycerol. The activity of glycerol kinase (EC 2.7.1.30) in leukocytes and cultured fibroblasts was less than 1% of the value for healthy subjects. PMID- 6299617 TI - Breast-feeding in industrialized and agricultural countries. PMID- 6299619 TI - The effect of iodized oil on goitre size, thyroid function and the development of the Jod Basedow phenomenon. PMID- 6299618 TI - Plasma steroid concentrations in patients with hypopituitarism and Kallman's syndrome: effects of testosterone replacement therapy. PMID- 6299620 TI - Opiate modulation of the pituitary-adrenal axis: effects of stress and circadian rhythm. AB - The opiate control of the pituitary-adrenal axis has been investigated in normal subjects. The infusion of 1 mg of the met-enkephalin analogue, DAMME, led to a fall in circulating cortisol in spite of a fall in blood pressure. Conversley, 16 mg of the opiate antagonist, naloxone led to brisk and pronounced elevations in plasma ACTH, lipotrophin (LPH) and cortisol. The rise above basal levels was consistent, irrespective of whether the infusion was given at 09.00, 18.00, or 23.00 h; the peak response obtained was significantly less at 23.00 h than at either 09.00 or 18.00 h. Finally, insulin-induced hypoglycaemia (0.15 u/kg) or naloxone (25 mg) produced a similar rise in plasma cortisol which was no different when the two stimuli were combined. It is suggested that there is a constant tonic inhibition of the pituitary-adrenal axis by endogenous opiates throughout 24 h, and that the circadian rhythm of ACTH/LPH secretion is not due to changes in opiate tone. However, disinhibition of this tone is likely to be responsible, at least in part, for the rise in cortisol in response to hypoglycaemic stress. PMID- 6299621 TI - Opiates control ACTH through a noradrenergic mechanism. PMID- 6299622 TI - Effects of TSH on cAMP levels and thyroid hormone release in human thyroid 'autonomous' nodules: relationship with iodothyronine and iodine content in thyroglobulin. AB - The effect of TSH on the adenylate cyclase-cAMP system and in vitro iodothyronine release, together with the iodothyronine and iodine content of 19s thyroglobulin, were studied in seven clinically euthyroid patients with autonomous thyroid nodules. Basal cAMP and cGMP content together with phosphodiesterase and protein kinase activities were normal in nodular, and suppressed in extranodular tissue. TSH-dependent cAMP accumulation was reduced in nodular tissue, but normal in the suppressed extranodular tissue. In vitro TSH-dependent iodothyronine release from nodular and extranodular tissue was absent. Thyroxine and iodine content of thyroglobulin extracted from nodular tissue was reduced, while triiodothyronine content was normal but with a low T4/T3 ratio. In extranodular tissue T3, T4 and iodine contents were reduced. In conclusion, autonomous thyroid nodules produced a poorly iodinated thyroglobulin leading to preferential T3 secretion with increased circulating free thyroid hormones even in clinically euthyroid patients. PMID- 6299623 TI - A comparison of three glucocorticoid suppressive regimes in adults with congenital adrenal hyperplasia. AB - We have compared three glucocorticoids, hydrocortisone (HC) (20 mg mane & 10 mg nocte), cortisone acetate (CA) (25 mg mane & 12.5 mg nocte), dexamethasone (DXM) (0.5 mg mane & 0.25 mg nocte), for their effect on the biochemical control of adult patients with congenital adrenal hyperplasia (CAH). Twenty-four-hour profiles of plasma concentrations of ACTH, 17-hydroxyprogesterone (170HP) and androstenedione (delta 4A), and 09.00 h dehydroepiandrosterone sulphate (DHAS) plasma concentrations were used to assess control. The patients were studied after 2 weeks on each glucocorticoid. The areas under the curves, the heights of the morning peaks of each hormone, the midnight concentrations, and the concentrations of hormones just before the evening dose were analysed. The results show that all the indices, except the midnight concentrations which were uniformly low, were significantly lower on DXM than on either HC or CA. There were no significant differences between HC and CA for any of the indices. The DHAS concentrations were low on all three glucocorticoids but again significantly lower on DXM. DXM (0.5 mg mane & 0.25 mg nocte) is therefore, in the short term, a better suppressor of the pituitary-adrenal axis in adults with CAH than either HC or CA, and in the dose used did not suppress ACTH to undetectable levels, nor the steroids to below levels found in normal subjects. PMID- 6299624 TI - Trilostane and the normal hypothalamic-pituitary-adrenocortical axis. AB - Trilostane has been used to treat Cushing's syndrome and other adrenocortical disorders. To investigate its effect on the normal adrenal gland, trilostane (initially 240 mg/day) was given to ten healthy men, the dose increasing at weekly intervals by 240 mg/day up to a final dose of 960 mg/day. The drug was well tolerated although one subject withdrew after the first week because of gastrointestinal side effects. Trilostane had no significant effect on aldosterone levels or blood pressure. The mean 24-h urinary free cortisol excretion rose from 14.2 to 22.0 mumol/mol creatinine (P less than 0.01) before and after trilostane 240 mg/day but did not rise thereafter. Early morning serum cortisol and plasma ACTH levels did not change on trilostane. The mean increment in serum cortisol after the i.v. injection of 0.25 mg of ACTH was reduced from 398 nmol/l before trilostane to 287 nmol/l on 240 mg/day and to 291 nmol/l on 960 mg/day (P less than 0.01). Insulin-induced hypoglycaemia while on 960 mg/day produced a maximum increment in serum cortisol of 361 +/- 118 nmol/l (mean +/- SD) although one subject had a subnormal increment of 180 nmol/l (normal greater than 200 nmol/l). Plasma ACTH rose with hypoglycaemia in all cases. We conclude that trilostane has only a minor effect on the normal hypothalamic-pituitary adrenocortical axis. PMID- 6299625 TI - Effect of cyproterone acetate-ethinyl oestradiol treatment on adrenal function in hirsute women. AB - Pituitary adrenal suppression has been assessed in twenty-two patients receiving cyproterone acetate (100 mg) and ethinyl oestradiol (30 micrograms) for the treatment of hirsutism. Cortisol responsiveness to ACTH and to insulin induced hypoglycaemia, and diurnal variations of plasma cortisol levels were measured before and after 6 months of treatment. There was no evidence that adrenal suppression is a significant problem in this group of patients. PMID- 6299626 TI - Stress-induced secretion of ACTH and corticosterone during development of spontaneous hypertension in rats. AB - Plasma ACTH and corticosterone response to ether inhalation was examined in spontaneously hypertensive rats (SHR) and in normotensive Wistar-Kyoto (WKY) and Wistar control rats at 4, 8, 12 and 16 weeks of age. In SHR, blood pressure rose to hypertensive levels (above 150 mm Hg) between 5 and 8 weeks, whereas, in control animals, it did not exceed 140 mm Hg. Maximum plasma ACTH concentrations were measured 5 min after exposure to ether and plasma corticosterone values attained their maximum at 20 min. Stress-induced ACTH release was significantly elevated in SHR at 4 and 8 weeks. At 16 weeks of age, no difference in the ACTH response between the three strains could be detected. Correspondingly, 4-week-old SHR showed a more pronounced corticosterone release than Wistar and WKY, whereas 16-week-old SHR and control rats responded similarly. Hypothalamo-pituitary adrenocortical response to ether stress is thus markedly enhanced in SHR during early development of hypertension. PMID- 6299627 TI - The baroreceptor-heart rate reflex in ACTH hypertension in sheep. AB - ACTH administration in sheep produces an adrenally dependent rise in blood pressure. Cardiac output and heart rate are usually increased. The precise mechanisms involved in the genesis of the hypertension are unclear. This study examines the sensitivity of the baroreflex heart rate response to phenylephrine hydrochloride and sodium nitroprusside before, during and after ACTH administration in sheep. During ACTH administration there was a sustained rise in blood pressure within 24 hours, whereas heart rate rose gradually. There was a sustained fall in baroreflex sensitivity to sodium nitroprusside within 24 hours, whereas baroreflex sensitivity to phenylephrine fell gradually over the first three days. The different time course of the change in sensitivity suggests that two different mechanisms are influenced by ACTH administration, for instance, changes in function in both cardiac vagal efferents and sympathetic pathways. PMID- 6299628 TI - Reduced sodium-potassium dependent ATPase and its possible role in the development of hypertension in spontaneously hypertensive rats. AB - Ouabain-sensitive Na+-K+-ATPase activity in red cell membranes, kidney cortical tissue, myocardium and adrenal glomerulosa tissue was examined in SHR and WKY rats at 6, 9, and 12 weeks of age. Red cell membrane enzyme activity was decreased (p less than 0.001) at 9 and 12 weeks of age in SHR. This activity was negatively correlated (r = -0.69, p less than .005) with blood pressure at 9 and 12 weeks. Kidney cortical enzyme activity was also decreased (p less than 0.001) in the SHR at 9 and 12 weeks of age. This decreased kidney enzyme activity was also inversely related to 9 and 12 week blood pressures (r = -0.71, p less than 0.001), urinary Na excretion (r = -0.62, p less than .005), and urinary Ca and K excretion. Myocardial enzyme activity was not decreased until 12 weeks in the SHR, and adrenal glomerulosa activity was not different in the SHR and WKY at any of the three ages that this enzyme was measured. Of the tissues examined decreased Na+-K+-ATPase activity in the erythrocyte membrane and in kidney cortical tissue appears to coincide best with the development of hypertension in the SHR. This study lends further support to the concept that alterations in membrane cation transport may be an important factor in the development of high blood pressure in SHR. PMID- 6299629 TI - Inhibition of angiotensin - converting enzyme in CBA mice with psychosocial hypertension. AB - Captopril (SQ 14,225) was acutely administered to control mice and to mice which had developed hypertension as a result of being caged in a manner which facilitates social interactions and competition. Systolic blood pressure was measured four hours after captopril administration, and blood was taken for measurement of plasma renin activity (PRA) several days later. Captopril produced no effect on normotensive control mice living in isolation nor did it reduce the blood pressure of the hypertensive mice on days 1, 7, and 14 following their placement in the special caging, despite the fact that their PRA was significantly elevated at these times. In contrast, by 1.5 months after placement in the cages and continuing for the next 5.5 months, captopril abolished the hypertension; PRA was normal during this period. We conclude that the increased PRA existing during the early stage of this psychosocial model of hypertension is not a significant direct contributor to the hypertension, whereas the renin angiotensin system may be essential for the maintenance of the hypertension beyond the early stage. PMID- 6299630 TI - Diuretic agents: actions on a molecular level. AB - The criteria upon which diuretics are classified are based upon their site of action within the nephron. Carbonic anhydrase inhibitors act in the proximal tubule, high-ceiling diuretics in the ascending loop of Henle, the thiazides in the early distal tubule and the potassium-sparing diuretics in the late distal tubule and in the collecting duct. According to the localization of carbonic anhydrase acetazolamide acts on three different sites in the proximal tubule cells. The loop diuretics inhibit the secondary active chloride reabsorption. Experiments on the isolated stripped rabbit colon under the condition of stimulated chloride secretion reveal striking similarities between the receptors for chloride reabsorption in the luminal cell membranes of the ascending loop of Henle and in the serosal cell membranes of the colon. The potassium-sparing diuretics act by blocking sodium channels in the distal parts of the nephron. The lumen negative potential difference decreases and potassium secretion is diminished. PMID- 6299631 TI - [Standardization for evaluation of plasma cyclic AMP response in Ellsworth-Howard test]. PMID- 6299632 TI - Glomus tympanicum tumours: demonstration by high resolution C.T. AB - The situation and extent of small glomus tumours confined to the middle ear cavity have been reliably demonstrated by high resolution C.T. The distinctive clinical and radiological features of four cases are described. High resolution C.T. is now considered to be the definitive radiological investigation and angiography is regarded as an unnecessary procedure in most patients with this condition. PMID- 6299633 TI - Increased IgM and IgM immune complex-like material in the circulation of renal transplant recipients with primary cytomegalovirus infections. AB - Twelve of 60 consecutive adult recipients of cadaver kidney transplants had increased polyethyleneglycol (PEG) precipitable IgM immune complex-like material in their circulation in the first 4 months after transplantation. All 10 recipients with primary CMV and two of four with secondary CMV infections had significant elevations in PEG precipitable IgM that coincided with rises in their CMV antibody titres. Ultracentrifuge analysis demonstrated two peaks of PEG precipitable material with sedimentation rates of about 20S and 40S. Total IgM immunoglobulin levels also were increased in transplant recipients with CMV infections, but this was less specific and occurred in patients without CMV infections. The Clq binding assay, which is more sensitive for IgG than IgM containing complexes, was positive in only three of 10 patients with primary CMV and none of four with secondary CMV. Granular deposits of IgM, but not IgG, were detected in the glomeruli of six of seven transplants biopsied during CMV infection. The PEG-IgM assay was not influenced by rejection or prednisone therapy. Thus, transplant patients, who develop primary CMV infections, produce elevated levels of circulating IgM and IgM immune complex-like material. These findings may help to differentiate CMV infection from transplant rejection as well as to increase our understanding of the special pathogenic properties of CMV in transplant recipients. PMID- 6299634 TI - Effects of deoxyadenosine on the activation of ADA inhibited T & B cells. AB - Using an in vitro model of adenosine deaminase deficiency, we have shown that the triggering of T lymphocyte proliferation by PHA is inhibited by concentrations of deoxyadenosine likely to occur in the plasma of ADA deficient children with severe combined immunodeficiency (ADA-SCID). Prostaglandin E1 increases the sensitivity of T cells to this inhibition, suggesting that derangement of cyclic nucleotide metabolism underlies the profound T lymphopaenia seen in ADA-SCID and in patients treated with the ADA inhibitor, deoxycoformycin. The proliferation and Ig synthesis of ADA inhibited B cells stimulated with protein A and EBV is in general less sensitive to the inhibitory effects of deoxyadenosine. However, this sensitivity varies widely between experiments suggesting that other cells present in the cultures may modify the inhibitory effect of deoxyadenosine. This may help explain the variability in B cell numbers and function in patients with ADA-SCID. PMID- 6299635 TI - Activation of human basophils by staphylococcal protein A. I. The role of cyclic AMP, arachidonic acid metabolites, microtubules and microfilaments. AB - Protein A from Staphylococcus aureus (Staph A) induces histamine secretion from human basophil leucocytes in the concentration range 10(-4) - 10 micrograms/ml. This reaction has great similarities to that of antigen or anti-IgE-induced release. It is characterized by a two stage reaction, requires extracellular calcium and is optimal at 37 degrees C. The rate of release is similar to that of IgE-mediated reactions. Histamine release induced by Staph A is inhibited by metabolic inhibitors, drugs which increase intracellular cyclic AMP levels, inhibitors of lipoxygenase pathways and a phospholipase A2 inhibitor. D2O and cytochalasin B which affect microtubules and microfilaments respectively, enhance histamine release induced by Staph A. These results suggest that Staph A-induced release is modulated by intracellular cyclic AMP, arachidonic acid metabolites, requires energy and is enhanced by the disruption of microfilaments and stabilization of microtubules. PMID- 6299636 TI - Specific and non-specific B cell activation in measles and varicella. AB - Lymphocytes from eight patients with measles and six patients with varicella were studied during the acute phase (first week) of illness and after recovery for spontaneous and pokeweed mitogen (PWM)-induced production of immunoglobulins (Ig) and viral antibodies by an enzyme linked immunosorbent assay (ELISA). In both infections acute phase lymphocytes showed increased spontaneous in vitro IgM and IgG productions including IgM and IgG antibodies to the aetiological virus as well as IgG antibodies to unrelated viruses (varicella, measles, rubella and mumps) to which the patient had serum antibodies. PWM induced no further Ig synthesis in the acute phase. In the convalescent phase viral antibody production could be demonstrated only in PWM stimulated cultures. In four patients the spontaneous synthesis of antibodies to a non-aetiological virus seemed to precede the production of IgG antibodies to the aetiological virus. All patients showed an increase of ELISA determined serum antibodies to the aetiological virus from the acute to the convalescent phase. Three of seven measles patients also showed a minor but significant increase or decrease of serum IgG antibodies to varicella and one of six varicella patients a significant rise of serum IgG antibodies to measles. Thus both measles and varicella infections were associated with non specific as well as specific B cell activation. The non-specific B cell activation may be induced by non-specific helper factors from activated T cells. PMID- 6299637 TI - Low beta-adrenergic receptor concentration on human thymocytes. AB - Several agents are known that can elevate cyclic AMP levels in lymphoid cells, e.g. isoproterenol, PGE1 and adenosine. We have studied the cyclic AMP increasing effect of these agents on thymocytes from mouse and man and on human peripheral T lymphocytes. In contrast to mouse thymocytes and human peripheral T lymphocytes, human thymocytes appeared to be insensitive to isoproterenol, but did respond to PGE1 and adenosine. Furthermore, the density of beta-adrenergic receptors on the cells was determined by measuring the specific binding of 3H-dihydroalprenolol. A correlation was found between the receptor density on the cells and the rise in intracellular cyclic AMP induced by isoproterenol: human thymocytes appeared to have very few beta-adrenergic receptors, in contrast to thymocytes from mouse or to T lymphocytes from human blood. We conclude that the development of beta adrenergic receptors in T cell ontogeny is different for mice and human beings. Comparison of animal models with the situation in man should be made with caution. PMID- 6299638 TI - Further studies on the effect of adenosine cyclic monophosphate derivatives on cell proliferation in the jejunal crypts of rat. AB - 1. Cell proliferation in the jejunal crypt epithelium of rat was measured using a stathmokinetic technique. 2. Sodium butyrate was found to promote jejunal crypt cell proliferation. 3. N6, O2'-Dibutyryl cyclic adenosine monophosphate (cAMP), N6-monobutyryl-cAMP and N6-monobutyryl-8-bromo-cAMP were found to inhibit cell proliferation when compared to sodium butyrate treated tissues. 4. 8 Chlorophenylthio-cAMP was found to inhibit cell division when compared to untreated animals. 5. O2'-Monobutyryl cAMP and 8-bromo-cAMP were not found to inhibit cell proliferation. PMID- 6299640 TI - Radionuclide demonstration of acute hemorrhage into follicular adenoma of the thyroid. PMID- 6299639 TI - Diseases caused by antibodies to receptors. PMID- 6299641 TI - Evidence for cardiac beta 2-adrenoceptors in man. AB - We compared the effects of single doses of 50 mg atenolol (cardioselective), 40 mg propranolol (nonselective), and placebo on both exercise- and isoproterenol induced tachycardia in two experiments involving nine normal subjects. Maximal exercise heart rate was reduced from 187 +/- 4(SEM) after placebo to 146 +/- 7 bpm after atenolol and 138 +/- 6 bpm after propranolol, but there were no differences between the drugs. The effects on isoproterenol tachycardia were determined before and after atropine (0.04 mg/kg IV). Isoproterenol sensitivity was determined as the intravenous dose that increased heart rate by 25 bpm (CD25) and this was increased from 1.8 +/- 0.3 micrograms after placebo to 38.9 +/- 8.3 micrograms after propranolol and 8.3 +/- 1.7 micrograms after atenolol. The difference in the effects of the two was significant. After atropine the CD25 was unchanged after placebo (2.3 +/- 0.3 micrograms) and atenolol (7.7 +/- 1.3 micrograms); it was reduced after propranolol (24.8 +/- 5.0 micrograms), but remained different from atenolol. This change with propranolol sensitivity was calculated as the apparent Ka, this was unchanged by atropine (11.7 +/- 2.1 and 10.1 +/- 2.5 ml/ng). These data are consistent with the hypothesis that exercise induced tachycardia results largely from beta 1-receptor activation that is blocked by both cardioselective and nonselective drugs, whereas isoproterenol activates both beta 1- and beta 2-receptors so that after cardioselective blockade there remains a beta 2-component that can be blocked with a nonselective drug. While there appear to be beta 2-receptors in the human heart, their physiologic or pathologic roles remain to be defined. PMID- 6299642 TI - Ranitidine bioavailability and kinetics in normal male subjects. AB - Ranitidine is a potent histamine H2-receptor blocker that inhibits histamine- and pentagastrin-induced gastric acid secretion. After doses of 100 mg both intravenously and orally ranitidine kinetics and bioavailability were investigated in a single dose two-way crossover study in 12 normal men. Serum concentrations of ranitidine were determined by radioimmunoassay and urine concentrations by an ion-pair HPLC method. Intravenous data were fitted to exponential equations with the computer program NONLIN; model-independent kinetic parameters were calculated. Elimination t 1/2, plasma clearance, renal clearance, hepatic clearance, and volume of distribution for ranitidine after intravenous injection were 2 hr, 10.4 ml/(min X kg), 7.2 ml/(min X kg), 3.1 ml/(min X kg), and 1.82 l/kg, respectively; after oral doses mean t 1/2 was 2.7 hr and mean bioavailability was 52%. The average cumulative urinary excretion of ranitidine as percent of dose was 69.4 +/- 6.1% and 26.7 +/- 7.2% after intravenous and oral doses. PMID- 6299643 TI - Oesophageal transit of a radionuclide solid bolus in normals. AB - The purpose of this study was to evaluate the physiological transit pattern of a compact bolus through the oesophagus in normal subjects. A standard gelatine bolus labelled with 99mTechnetium pertechnetate was swallowed in the erect and the supine positions and traced by a gamma-camera in 20 subjects. In the erect position, the mean overall transit time was 5.5 +/- 1.1 s (SD) which differed significantly (P less than 0.001) from the longer mean overall transit time of 6.9 +/- 1.4 s (SD) in the supine position. The transit patterns differed between erect and supine positions. In the erect position, the bolus travelled at a constant velocity until the most distal part of the oesophagus where a slight retardation of the velocity occurred, whilst in the supine position, a uniform retardation of the velocity was seen which was slightly accentuated in the most distal part. The present data provide a basis for the evaluation of studies on the bolus transport in patients with suspected oesophageal dysfunction. PMID- 6299644 TI - Malignant and benign brain tumors: current concepts and intervention. PMID- 6299645 TI - Germ cell tumors of the testis. PMID- 6299646 TI - Techniques and approaches to proton NMR imaging of the head. AB - The next few years will undoubtedly see a refinement of proton imaging technology and a broader data base will indicate to what extent proton relaxation parameters are able to detect and characterize disease. In addition, it is likely that imaging of other nuclei (e.g. 31P, 23Na, 19F) will become a reality, although it must be stated that due to their inherently lower sensitivity to NMR detection and/or lower physiological concentration, clinical images of nuclei other than 1H will undoubtedly have a low spatial resolution and may require relatively long imaging times [41]. Nonetheless, herein lies the exciting possibility of non invasive metabolic or functional imaging [42]. The realm of NMR contrast agents is just beginning to be explored [43, 44], and developments in high-speed imaging [45] indicate useful applications in cardiology [46]. So whilst improvements in image quality can be expected, as was the case with X-ray CT, the application of NMR in medicine will diversify to yield information of a more specifically functional nature. This, together with the very low attendant biological risk [47], heralds a bright future for NMR in clinical diagnosis. PMID- 6299647 TI - Sulfated galactosaminoglycans of bovine periodontal ligament. Evidence for the presence of two major types of hybrids but no chondroitin sulfate. AB - Sulfated galactosaminoglycans of mature bovine periodontal ligament were separated into four fractions by ethanol precipitation. Fractions I and II were dermatan sulfates with high contents of L-iduronate, but only small amounts of this hexuronic acid were present in fractions III and IV. Effects of digestion with testicular hyaluronidase or a periodate-alkali treatment showed that most if not all of the glycans in fractions I, II and III were hybrid chains containing both L-iduronate and D-glucuronate. The composition of fraction IV was less certain, but the chains strongly resembled fraction III hybrids in electrophoretic characteristics, not chondroitin sulfate. The total amount of the D-glucuronate-rich fractions III and IV in the ligament was similar to that of I plus II. In contrast, almost all of the sulfated galactosaminoglycans of mature skin were rich in L-iduronate. The more varied composition of the ligament glycosaminoglycans may be related to the mixed population of cells in this tissue. PMID- 6299649 TI - Diagnosis of liver tumors by dynamic computed tomography. PMID- 6299650 TI - Sodium/potassium ATPase in normal and cataractous human lenses. AB - Na/K ATPase activity has been determined in normal and cataractous human lenses using labeled ATP as a substrate. The enzyme is distributed throughout the lens with approximately 1/3 of the total activity in the capsule-epithelium and 1/2 of the activity in the cortex. Furthermore, the activity of Na/K ATPase decreases with increasing age and this decrease occurs primarily in the inner nuclear region. In severe cataractous lenses, a marked decrease in the Na/K ATPase activity has been demonstrated in all parts of the lens, in contrast to immature cataracts, where the decrease in the enzyme activity occurs primarily in the cortical and nuclear regions. PMID- 6299648 TI - Mechanism of crosslinking of proteins by glutaraldehyde II. Reaction with monomeric and polymeric collagen. AB - Collagen in three different states, i.e. native soluble molecules, denatured molecules and reconstituted fibers, was exposed to various concentrations of glutaraldehyde. The degree of intramolecular and intermolecular crosslink formation was evaluated by measuring collagen solubility, beta and gamma chain formation, resistance towards cleavage by CNBr or collagenase digestion. Modification of lysyl residues was measured by amino acid analysis. When dilute collagen solutions were reacted with low concentrations of glutaraldehyde, intramolecular crosslinks were observed as the predominant crosslinks. When the glutaraldehyde concentration was increased, the collagen became more insoluble, indicating the formation of intermolecular crosslinks. When reconstituted collagen fibers were reacted with low concentrations of glutaraldehyde, intermolecular crosslinks were formed, which prevented the material from being solubilized by CNBr. However, these materials could still be solubilized by collagenase. When the glutaraldehyde concentration was increased, the materials became resistant to collagenase, while there was only a small increase in the number of lysyl residues modified. This reflects an increase in the molecular length of the glutaraldehyde polymers extending from the initial glutaraldehyde and lysyl residue reaction sites rather than an increase in the actual number of crosslinking sites. PMID- 6299651 TI - Surgical treatment of farmyard pox. Orf, milker's nodules, bovine papular stomatitis pox. AB - Superficial epidermal surgical removal of localized bullous lesions of viral origin is recommended. An example is presented in which the patient had what we have termed "farmyard pox." This is our generic label for the clinically indistinguishable parapox viral infections acquired from farm animals, which include orf, milker's nodules, and the pox of bovine papular stomatitis. The surgery is simple and rapid, and completely removes the lesions. This eliminates the possibility of enlargement and contagion, and also promotes rapid healing. PMID- 6299652 TI - A multicenter double-blind evaluation of ketoconazole in the treatment of dermatomycoses. AB - A multicenter double-blind study was conducted on the use of ketoconazole and griseofulvin for the treatment of dermatomycoses. Of one hundred thirty cases (one hundred twenty-seven patients) for which efficacy data were available, sixty six were treated with a single daily dose of 200 mg ketoconazole, and sixty-four were treated with a single daily dose of 250 mg griseofulvin for periods of two to sixteen weeks. The proportion of remissions observed with ketoconazole (61 percent) was significantly greater (p = 0.02) than that observed with griseofulvin (39 percent). The proportion of relapses within two months was significantly less (p less than 0.01) in the ketoconazole group (9 percent) than in the griseofulvin group (43 percent). The frequency and severity of side effects were comparable in the two groups. PMID- 6299653 TI - The Danish testicular carcinoma study (DATECA). The DATECA Working Group. PMID- 6299654 TI - Angiotensin-converting enzyme in human adult respiratory distress syndrome. AB - Angiotensin-converting enzyme (ACE) levels, complement activation, and intravascular coagulation were studied in 36 patients with adult respiratory distress syndrome (ARDS) (17 aseptic, 19 septic), in order to investigate the possible interrelations among ACE, immunologic data, and hematologic findings. The severity of respiratory impairment was assessed with measurements of mechanical and gas exchange functional qualities of the lung. Serial measurements of ACE could be done in 14 patients during an eight-day period. During the first 24 hours, ACE levels were always normal (38 percent) or decreased (62 percent). No difference could be found between patients with septic and aseptic ARDS. Complement activation occurred in 78 percent (28/36) and used, in most cases, the classic pathway with presence of circulating immune complexes. Criteria for intravascular coagulation were present in 58 percent (21/36). No relation between coagulation, complement, and ACE could be found except for the patients with a greater respiratory impairment, who had complement activation, intravascular coagulation, and significantly lower ACE levels. In all patients together, ACE levels had no diagnostic value for aseptic cause of ARDS and a poor prognostic value. Only intravascular coagulation was linked with a higher significant mortality and a greater functional impairment. Serial measurements showed a diphasic evolution of ACE levels, with a maximum decrease between the 72nd and 96th hours and a further normalization (seventh day). The persistence of low levels seemed to be associated with evolutive sepsis or secondary aggravation and fibrosis. PMID- 6299655 TI - Intravenous bicarbonate may cause transient intracellular acidosis. PMID- 6299656 TI - Elimination kinetics of cefotaxime and desacetyl cefotaxime in patients with renal insufficiency and during hemodialysis. AB - The pharmacokinetics of cefotaxime, a new semi-synthetic cephalosporin for injection, was studied in 30 subjects with various degrees of renal function after a single 1-gram intramuscular injection. Serum and urinary concentrations of cefotaxime and desacetyl cefotaxime were determined by high pressure liquid chromatography. The pharmacokinetic parameters of cefotaxime were obtained using a one-compartment open model. The mean serum half-life of the parent compound (cefotaxime), 0.87 h in normal subjects, was prolonged to 2.35 h in hemodialysis patients. There was a significant linear correlation between the elimination rate constant of cefotaxime and creatinine clearance. The mean cumulative urinary recovery of the administered dose in the 24-hour urine was 51.7% as cefotaxime and 25.6% as desacetyl cefotaxime in normal subjects. PMID- 6299657 TI - Acupuncture in acute myocardial infarction. PMID- 6299659 TI - Enteritis necroticans (Pigbel) an unrecognised preventable disease? PMID- 6299658 TI - Insulin secretion and metabolic changes in maturity onset diabetes mellitus and glucose intolerance. PMID- 6299660 TI - Acupuncture channels--myth or reality. PMID- 6299661 TI - [Investigation of the incidence of delayed silicosis in 2 mines with different contents of free silica]. PMID- 6299662 TI - Nucleotide sequence of a human immunoglobulin C gamma 4 gene. AB - We report the nucleotide sequence of a gene encoding the constant region of a human immunoglobulin gamma 4 heavy chain (C gamma 4). These data represent the first complete sequence determination of a human CH gene. As expected from structural studies of mouse C gamma genes, the coding sequences for the CH domains and hinge segment are separated from one another by intervening DNA sequences. Comparison with genomic sequences of the mouse C gamma 1, C gamma 2a, and C gamma 2b genes shows conservation of the sequences in the constant region domains and the 3' untranslated region surrounding the presumed site of poly(A) addition. PMID- 6299663 TI - Hormonally regulated mammalian gene expression: steady-state level and nucleotide sequence of rabbit uteroglobin mRNA. AB - Uteroglobin is a protein that is synthesized in large quantities by the rabbit uterine endometrial cells and secreted into the uterine lumen around the time of implantation of the developing blastocysts. The protein is also synthesized constitutively at a low level in the lung. In the uterus, synthesis of the protein is induced by progesterone but repressed by estradiol; whereas in the lung, it is not hormonally responsive. Using a full-length cDNA clone, we have established the nucleotide sequence of uteroglobin mRNA and have determined its levels in uterus and lung during early pregnancy. The clone, pUG617, contains all but 24 nucleotides at the 5' untranslated region of the structural gene. To establish the full mRNA sequence, we isolated a 5' end-labeled DNA fragment from pUG617 and extended its length using reverse transcriptase after hybridization with uterine poly(A)-containing RNA. The 5'-terminal sequence of uteroglobin mRNA was established by sequencing the extended DNA fragment. The nucleotide sequence of the peptide-coding portion of the gene has resolved some previously reported discrepancies in the amino acid sequence of the mature protein and those in the signal peptide. By comparison of sequences with a partial uteroglobin cDNA clone isolated by another laboratory, a polymorphic nucleotide at position 246 of the gene has been identified, where a G-A transition has caused an amino acid substitution from aspartic acid to asparagine at residue 46 of the mature protein. Analysis of steady-state RNA levels in the uterus has shown that the induction and repression of uteroglobin synthesis during early pregnancy is the result of accumulation and depletion of its mRNA, respectively. During the same period in the lung, no consistent changes in uteroglobin mRNA level were evident, reflecting the constitutive levels of the protein in this tissue. PMID- 6299664 TI - Plasmid vectors capable of transferring large DNA fragments to yeast. AB - We have constructed several cloning vectors which can be used in vitro packaging and yeast transformation. These plasmids have been designed for the convenient cloning of large segments of DNA and their transfer to yeast. They contain bacterial plasmid DNA sequences for replication and selection in Escherichia coli, yeast 2-microns plasmid DNA sequences or chromosomal replicators and yeast markers necessary for replication and selection in yeast, and the cohesive ends of bacteriophage lambda which allow packaging of recombinant molecules into lambda phage heads. Large fragments (22-38 kb) of Klebsiella pneumoniae and Zea mays DNA were ligated into plasmid vector pBTI-1 to make complete genome libraries. One clone from the K. pneumoniae library was amplified in E. coli and the purified DNA used to transform yeast cells. Transformation of yeast by large DNA fragments occurred at high frequencies. The recombinant plasmid was stably maintained in yeast, provided selective pressure for Leu+ transformants was maintained. The structurally complete recombinant plasmid can be recovered from yeast by transforming E. coli to ampicillin resistance. Fewer than 5% of the recovered plasmids had undergone recombination with endogenous yeast 2-microns plasmid. PMID- 6299665 TI - Cloning of bovine prolactin cDNA and evolutionary implications of its sequence. AB - Prolactin, growth hormone, and chorionic somatomammotropin (placental lactogen) constitute a set of related polypeptides believed to derive from a common evolutionary ancestor protein. We have cloned and sequenced DNA complementary to the mRNA coding for bovine prolactin. This cDNA contains 702 bases corresponding to 10 amino acids in the leader peptide, all 199 amino acids of the hormone, and 75 nucleotides in the 3' untranslated region of the mRNA. Nucleotide sequence analysis of this cDNA permitted the identification of 10 amino acids in the signal peptide, plus the correction or elucidation of amino acid assignments at 16 sites where aspartic and glutamic acids had not been distinguished from their amides by amino acid sequencing. Codon usage in bovine prolactin mRNA is nonrandom, but, similarly to rat and human prolactins, it does not exhibit the strong preference for G or C in codon third positions seen in bovine, rat, and human growth hormone mRNAs. The translational termination signal in bovine prolactin in UAA, also the same as in rat and human prolactins and differing from the UAG "stop" codon used in bovine, rat, and human growth hormones and human chorionic somatomammotropin. The amino acid and mRNA nucleotide sequences of bovine, rat, and human prolactins and growth hormones were compared by several techniques based on various theories of molecular evolution. The comparison of prolactin to growth hormone is consistent in all three species, suggesting that the genes for these two hormones diverged about 350 million years ago. However, comparisons among the three prolactins or among the three growth hormones to determine the times of evolutionary divergence of the three species generated values that were inconsistent with each other and with the fossil record. Analysis of these discrepancies suggests that the genes for prolactin and growth hormone may now be evolving by different mechanisms. PMID- 6299666 TI - Cloning and sequencing of restriction fragments generated by Eco RI*. AB - Thirty-four Eco RI* sites have been identified on the nucleotide sequence of CaMV, following cloning of Eco RI* fragments in M13mp2. From this sequencing data, we have deduced that Eco RI* recognizes sites that differ in a single position from the canonical Eco RI sequence, GAATTC. Any substitution can occur at any one of the six positions in the recognition site, with the exception of A leads to T or T leads to A changes within the central tetramer. The Eco RI* restriction patterns of phi x174 and pBR322 are consistent with these recognition criteria. Similarly, Bam HI* cleavage of phi x174 and SV40 (George et al., 1980) produces restriction patterns that are consistent with single-position degeneracy in the canonical Bam HI recognition site. Cohesive termini produced by Eco RI* cleavage were ligated into the Eco RI site of M13mp2, even when there was a base pair mismatch within the four nucleotide overlap. Mismatches were corrected asymmetrically during subsequent replication of M13 in E. coli. PMID- 6299667 TI - Eco RI* specificity and hydrogen bonding. AB - Under standard conditions, Eco RI endonuclease uniquely recognizes the inverted repeat GAATTC. However, this specificity breaks down under non-standard conditions into what has been termed Eco RI* specificity, wherein many other sequences are recognized. We show here that the hydrolysis rates at all known Eco RI* sites can be summarized by the hierarchies: G much greater than A greater than T much greater than C at the first position, A much greater than [G,C] much greater than T at the second and third position, and the corresponding complements at the last three positions. This is consistent with a recognition model which assumes that there are two specific hydrogen bonds per base pair under standard conditions. One or more of these are randomly replaced by water under Eco RI* conditions and the position of a sequence within the appropriate hierarchy is primarily determined by the number of retained hydrogen bonds. These retained hydrogen bonds are common recognition features that can be identified by examining the DNA. The recognition points thereby identified for Eco RI all fall within the major groove of the DNA. PMID- 6299669 TI - [A modified 131 I immuno-autoradiographic method for detection of serum EB virus VCA-IgA antibody in nasopharyngeal carcinoma]. PMID- 6299668 TI - The human pro-opiomelanocortin gene: organization, sequence, and interspersion with repetitive DNA. AB - The human pro-opiomelanocortin (POMC) gene has been characterized by molecular cloning and DNA sequence analysis. Although this gene codes for several different polypeptide hormones, only a single intron interrupts the protein coding region. The DNA in this intron, and in a second intron found in the region of the gene homologous to the mRNA 5'-untranslated sequence, contains repetitive DNA sequences. At least some of these sequences belong to the Alu family of transcribed middle repetitive DNA. The determination of the complete nucleotide sequence of the coding regions of the gene demonstrates that the pattern of homologous and variable regions seen in the POMC protein between different species is reflected at the DNA level. DNA sequences encoding the highly conserved regions of POMC are 90-95% homologous between species while the coding sequences for the variable regions of the protein are approximately 70% homologous. The very high degree of homology in the amino terminal portion of POMC is consistent with an important physiological role for this peptide. PMID- 6299671 TI - [Storage and analysis of data from 46 cases of small hepatocellular carcinoma using microcomputer]. PMID- 6299670 TI - [A light and electron microscopic study of chemodectoma of the lung]. PMID- 6299672 TI - Hepatocellular carcinoma associated with oral contraceptive use and pregnancy. AB - Numerous undesirable side effects have been attributed to oral contraception, from mild breast discomfort to thromboembolism. The authors present a case report of hepatocellular carcinoma associated with oral contraceptive use and pregnancy and discuss the potentially fatal association of malignant liver tumors with usage of oral contraception. PMID- 6299673 TI - Histologic evidence for an association of cervical intraepithelial neoplasia with human papilloma virus infection. AB - All the cytologic and histologic material pertaining to 100 patients who underwent cervical conization for advanced cervical intraepithelial neoplasia (CIN) was reviewed. The revision of the histology of the biopsies and cones showed in 56 cases the association of CIN with viral cytopathic effects (VCE) attributable to human pappiloma virus (HPV) and in 52 the coexistence of a predominantly flat condyloma. The comparison of the two groups of CIN, with and without VCE, showed that in the first the association had favored in 20% of the cases the histologic overestimation of the severity of the lesion. Of the patients with CIN III, 46% showed additional changes due to VCE. The mean age of the patients with CIN and VCE was 39.8 years and that of the patients with CIN was 48.6 (p less than 0.0001). The exocervix was significantly more often involved by CIN + VCE than by CIN alone (p less than 0.00001). Follow-up studies revealed in both groups the same percentage of residual disease and, preliminarily, a trend to a better control of CIN with VCE. New disease developed more often in the group of patients with CIN without VCE. Cytologic sensitivity for VCE in cervical smears was high (95%) in the cases of CIN II and somewhat lower (81%) in those with CIN III. Cytologic follow-up showed the persistence of VCE in 17% of the patients treated surgically for CIN and VCE. The morphologic and clinical features displayed by CIN associated with VCE warrant its recognition as a distinct variant of CIN. PMID- 6299674 TI - Cytological diagnosis of virus-infected cells in cervical smears. Value in gynecologic and obstetric practice. AB - Herpes simplex virus is a common cause of infection of the female genital tract. The infection can be diagnosed by the cytopathologist from the appearances of multinucleate giant cells in Papanicolaou smears. The cytological diagnosis of this infection is of value to the gynecologist in identifying subclinical infection and in confirming clinically suspicious disease. The validity of the cytological diagnosis can be established by virus isolation or electron microscopy. Although other infections of the female genital tract such as wart virus infection, cytomegalovirus infection, and chlamydial infection can be detected by cytological methods, the reliability of the cytological approach has yet to be established. Nevertheless, cytology has provided a powerful stimulus for research into the prevalence of wart virus infection of the cervix, which has been shown to be much more common than was previously supposed, and may be a factor in cervical carcinogenesis. Cytological examination of smears of urinary sediment has proven to be a reliable method of detecting human polyomavirus excretion in pregnancy. Active infection of the urinary tract occurs in 3.2% of pregnant women who represent an "at risk group" with altered immunological function. PMID- 6299675 TI - Frozen section diagnosis of cytomegalovirus infections. AB - We studied the standard frozen section as a diagnostic method for cytomegalovirus infection of the lung in bone marrow transplant recipients. Fifty-nine patients with cytomegalovirus pneumonia diagnosed by open lung biopsy were studied. The sensitivity of frozen section was compared with that of paraffin section, cytology of imprints prepared from the tissue sample, and viral culture. Fifteen percent of the cases were diagnosable at frozen section alone, and 44% by cytology alone. This confirms our previous study and indicates that frozen section unaided by more specific indentification methods provides low sensitivity even in cytomegalovirus-infected cells, which probably show the most easily identified cytopathic effects of any virus. This low sensitivity underscores the need for the additional sensitivity and specificity which may be contributed by immunolabeling techniques to rapid viral diagnosis. PMID- 6299676 TI - [Effect of nucleotides, amiloride and ions on active ion transport by the frog skin. A single-membrane model]. PMID- 6299677 TI - [Isolation of the transmembrane calcium flow of smooth muscle cells in a potassium-free medium]. PMID- 6299678 TI - [Phosphorylation of bound [14C] GDP in preparations of the frog retinal rod outer segments]. PMID- 6299679 TI - [2 types of calcium channels in the somatic membrane of spinal ganglion neurons in the rat]. PMID- 6299680 TI - Brain membrane disordering related to acute ethanol administration in naive and short-term ethanol-intoxicated rats. AB - Crude synaptic membrane fluidity (checked by fluorescence polarization) together with (Na+ + K+) ATPase activity were examined 18 hours after a single oral ethanol administration (5 g/kg bwt.) to naive rats and to rats previously intubated with ethanol repeatedly during 4 days. The sensibility of both parameters to different concentrations of ethanol added in vitro (0.175 M-1.400 M) was also determined. Although no changes in the basal intrinsic fluidity were found, (Na+ + K+)ATPase activity increased slightly in both conditions. The fluidizing as well as the ATPase inhibiting effects following the addition of ethanol in vitro were markedly increased 18 hours after ethanol administration to naive rats. This hypersensitization was no longer apparent in rats pretreated with ethanol during 4 days. The acute ethanol-induced hypersensitization found in naive rats appears not to be related to an unspecific stress or to changes in body temperature. The disappearance of this hypersensitization in short-term alcohol-intoxicated animals may represent the first stage of tolerance acquisition. PMID- 6299681 TI - Peripheral neuropathy after disulfiram administration: reversibility despite continued therapy. AB - The clinical history of an abstinent alcoholic developing paresthesias after one year's treatment with disulfiram (250 mg daily) is presented. Despite continued disulfiram in decreased dosage (125 mg daily), associated with multivitamin supplementation, marked improvement in symptoms and signs was noted, contrary to previous case reports. The peripheral sensorimotor neuropathy associated with disulfiram administration has been well described [1 - 8]. From those reports, it appears that symptoms may appear from several weeks to several months after the drug has been administered. Although the etiology of this neuropathy remains speculative, the usual treatment is to discontinue disulfiram once the toxicity develops. Presented here is the clinical course of a subject, whose neuropathic symptoms resolved with continued disulfiram administration at a lower daily dosage in conjunction with multivitamin therapy. PMID- 6299682 TI - The effect of chronically administered delta-9-tetrahydrocannabinol upon the polygraphically monitored sleep of normal volunteers. AB - This study describes the effect of two weeks of delta-9-tetrahydrocannabinol (THC) administration upon normal sleep. The two subjects, two brothers in their 20s, slept in the laboratory for 27 consecutive nights and then, after four nights at home, for four additional nights. One subject, after an adaption night, received placebo for four baseline nights, 30 mg of THC for the next 14 nights, and placebo during four withdrawal nights. The other subject received placebo during this entire period. One year later the subjects alternated these conditions. The subjects had difficulty falling and staying asleep during the first two nights of placebo after 14 consecutive drug nights. This mild drug withdrawal insomnia was not accompanied by the increase of REM sleep which frequently accompanies withdrawal of other drugs. Starting after about a week of THC administration, and continuing for a week after drug discontinuance, there was a marked decrease in the type of sleep associated with slow waves in the electroencephalogram, nonREM sleep stages 3 and 4. The fact that prolonged, but not acute use, suppresses slow wave sleep indicates that this commonly used drug produces a poorly understood change in brain physiology. PMID- 6299683 TI - [Effect of cimetidine and ranitidine on blood plasma coagulability]. PMID- 6299684 TI - [Correlations between reovirus infections and Salmonella typhimurium var. copenhagen infections in pigeons]. PMID- 6299685 TI - [Maternal antibodies against Aujeszky-virus in piglets of vaccinated sows]. PMID- 6299686 TI - An isolated case of juvenile angiofibroma in the nasal cavity. PMID- 6299688 TI - Serotonin and adrenocorticotropin (ACTH) release: direct effects at the anterior pituitary level and potentiation of arginine vasopressin-induced ACTH release. PMID- 6299687 TI - Gonadotropins regulate plasminogen activator production by rat granulosa cells. AB - The role of FSH in the regulation of plasminogen activator production was studied in granulosa cells obtained from 23- to 25-day-old female rats. Cells cultured without FSH secreted a negligible amount of plasminogen activator. Purified ovine, rat, and human FSH produced dose-dependent increases in plasminogen activator production. This FSH effect was mimicked by analogs of cAMP, prostaglandin E2, and choleratoxin. Purified ovine LH and ovine PRL had no effect on plasminogen activator production by these immature granulosa cells. However, when these granulosa cells were treated in vitro or in vivo with FSH and then exposed to LH or PRL, the cells responded to LH in a dose-dependent manner with increased plasminogen activator production. These cells remained unresponsive to PRL. Similarly to FSH, in vitro pretreatment of the cells with choleratoxin or analogs of cAMP also induced responsiveness to LH with increased plasminogen activator production. This responsiveness to both FSH and LH with increased plasminogen activator production was also observed in granulosa cells obtained from rat preovulatory follicles. These studies demonstrate that: 1) FSH but not LH regulates plasminogen activator production by immature granulosa cells from preantral follicles; 2) Pretreatment of the undifferentiated granulosa cells with FSH, choleratoxin, or cAMP induced granulosa cell responsiveness to LH with increased plasminogen activator production; and 3) Granulosa cells obtained from preovulatory follicles respond to both FSH and LH with increased plasminogen activator secretion. These results suggest that with the LH surge at ovulation, plasminogen activator production in follicles is increased and may be important in follicular rupture. PMID- 6299689 TI - Biological activities and receptor interactions of des-Leu16 salmon and des-Phe16 human calcitonin. AB - Analogs of salmon (des-Leu16 sCT) and human (des-Phe16 hCT) calcitonin were prepared in which the amino acid from position 16 was omitted. The biological activities were assessed in vivo in the rat hypocalcemic assay and in vitro by studying competition for binding of [125I]sCT and adenylate cyclase stimulation in human breast cancer cells (T 47D). Deletion from position 16 resulted in substantial loss of biological activity in each system, indicating the importance for a hydrophobic residue at position 16 in the intact calcitonin molecule. PMID- 6299690 TI - Binding of radioiodinated bovine parathyroid hormone-(1-84) to canine renal cortical membranes. AB - Binding of PTH to purified canine renal cortical membranes was investigated using biologically active radioiodinated bovine PTH-(1-84) [bPTH-(1-84)] as radioligand. PTH-(1-84) is thought to be the major circulating form of bioactive PTH, but oxidative inactivation upon iodination has prevented its use as a radioligand probe of PTH receptors. We have labeled PTH-(1-84) by a microelectrolytic constant current method to a high specific activity (180-220 muCi/micrograms), corresponding to an average ratio of 1 mol 125I/mol peptide. Affinity purification on chick renal membranes consistently improved the radioligand-binding properties, with a 6-fold increase in fraction specifically bound. Analysis of equilibrium (180 min at 15 C) competition curves showed two classes of binding sites for bPTH-(1-84). A high affinity binding site appeared to be coupled to activation of adenylate cyclase and exhibited affinity varying between 1.9 and 4.3 X 10(8) M-1. The affinity of this site for bPTH was decreased more than 50% by the nonhydrolyzable analog of GTP, guanyl-5'-yl imidodiphosphate. A low affinity binding site also was detected (Ka = 0.6-6.2 X 10(6) M-1), and its affinity for bPTH was modulated by the concentration of magnesium. The high affinity sites exhibited hormonal specificity and guanine nucleotide dependency characteristic of peptide hormone receptors, while the low affinity sites did not. Analysis by polyacrylamide gel electrophoresis or high pressure liquid chromatography of the radioligand incubated with or released from purified canine renal cortical membranes revealed a single peak of radioactivity that comigrated with [125I]iodo-bPTH-(1-84). It appears from these studies that cleavage of amino-terminal to residue 43 ([125I]tyrosine) was not required for binding or release of hormone. These experiments indicate that radioiodinated bPTH-(1-84) is a useful probe for further characterization of PTH receptors in kidney and other organs. PMID- 6299692 TI - Cyclic nucleotides can induce luteinizing hormone receptor in cultured granulosa cells. PMID- 6299691 TI - Acetylcholine and norepinephrine: compared actions thyroid metabolism. AB - Acetylcholine (ACh; 5 X 10(-4) M), like norepinephrine (NE; 6 X 10(-6) M), as shown previously, stimulated iodide organification by mouse thyroids in vitro, while at the same time it inhibited TSH- or (Bu)2cAMP-induced T4 release. However, thyroid cAMP was not changed by ACh, suggesting that ACh, like NE, exerted its effects at a step beyond cAMP production. Also, while ACh increased cGMP concentrations, (Bu)2cGMP and 8-bromo-cGMP were not effective on thyroid function in this system. Neurotransmitters, then, presumably do not exert their action through cyclic nucleotide stimulation ACh-induced stimulation of organification and inhibition of release was reversed by 10(-5) M atropine (ATR) but not by 10(-5) M d-tubocurarine, indicating that muscarinic receptors were involved. ATR also reversed inhibition of T4 release induced by NE, suggesting that the presynaptic cholinergic pathway may be responsible for stimulation of postsynaptic cholinergic and adrenergic neurotransmitters in the thyroid gland. PMID- 6299693 TI - Nicotinamide adenine dinucleotide kinase in the rat uterus: regulation by progesterone and decidual induction. PMID- 6299694 TI - Thyroid hormone increases type I adenosine 3', 5'-monophosphate-dependent protein kinase and casein kinase activities in rat liver cytosol: analysis of protein kinases by polyacrylamide disc gel electrophoresis. AB - We investigated the action of thyroid hormone on each protein kinase in rat liver cytosol. Kinases were analyzed by polyacrylamide disc gel electrophoresis and isoelectric focusing in polyacrylamide gel. Polyacrylamide disc gel electrophoresis separated cAMP-dependent protein kinase type I (Rf = 0.35), type II (Rf = 0.44), their catalytic subunit (Rf = 0.26), and cAMP-independent protein kinase (Rf = 0.50). Casein kinase was detected at Rf = 0.37. In addition to the catalytic subunit with Rf = 0.26, another catalytic subunit was found at Rf = 0.44 when the cytosol was preincubated with cAMP. The administration of T3 (20 micrograms/100 g BW for 3 days) to hypothyroid rats increased enzyme activities of type I holoenzyme and casein kinase by 48%. Free catalytic subunit, separated from holoenzyme, had the same level of enzyme activity in both groups, suggesting greater endogenous dissociation of type I holoenzyme in hypothyroid rats. When heat-inactivated rat liver cytosol was used as substrate in the assay of protein kinase activity, the peak enzyme active in phosphorylating the cytosol corresponded to the casein kinase peak. Our data indicate that casein kinase is the main enzyme that mediates phosphorylation of endogenous proteins in rat liver cytosol, and that T3 treatment increases the activity of casein kinase and of type I cAMP-dependent protein kinase. PMID- 6299695 TI - Seasonal changes in testicular gonadotropin receptors and steroid content in the ram. PMID- 6299696 TI - Changes in testicular gonadotropin receptors and steroid content through postnatal development until puberty in the lamb. PMID- 6299697 TI - Quantitation of 3,5,3'-triiodothyronine (T3) receptors by a microcentrifuge exchange assay: evidence for the existence of a nucleoplasmic pool of rat liver T3 receptors. PMID- 6299698 TI - Solubilization of the gonadotropin-releasing hormone receptor from bovine pituitary plasma membranes. AB - The plasma membrane receptor for GnRH in bovine anterior pituitaries has been solubilized with the cholic acid derivative detergent 3([3 cholamidopropyl)dimethylammonio]propane sulfonate. The soluble receptor in the supernatant from a 100,000 x g x 90 min centrifugation displays high affinity, saturability and specific binding to the agonist, [DA1a6,N alpha MeLeu7, Pro9 NEt]-GnRH. The dissociation constant, KD, for the soluble receptor is 0.56 +/- 0.07 nM (mean +/- SEM) and the number of sites, Ro, is 85 +/- 17 fmols/mg protein. For the membrane-bound receptor the KD is 0.50 +/- 0.04 nM and Ro is 300 +/- 18 fmols/mg protein. The relative potencies of GnRH and the potent antagonist, [Ac-delta 3Pro1,pFDPhe2, DTrp3,6]-GnRH are similar for both the soluble and membrane-bound receptor. PMID- 6299699 TI - The effect of ketoconazole on steroidogenesis in cultured mouse adrenal cortex tumor cells. PMID- 6299700 TI - ACTH stimulation of adrenal steroidogenesis is sensitive to trypsin inhibitors. PMID- 6299701 TI - Influence of exogenous melatonin on melatonin secretion and the neuroendocrine reproductive axis of intact male rats during sexual maturation. AB - The influence of daily sc administration of melatonin (5-100 micrograms/day) on sexual development of prepubertal and pubertal male rats was studied in vivo. Adult animals were also studied. When melatonin was injected daily into young animals starting at day 20 of age, dose-dependent reductions in plasma testosterone, testis and seminal vesicles weights, plasma FSH and LH levels, and pituitary GnRH receptor number were observed at day 40 or 45 of age when animals were killed. In contrast, prepubertal animals (5-20 days old) showed no significant responses to similar treatment with melatonin, whereas in adult animals (70-90 days old), melatonin elicited only a small decrease in plasma testosterone concentration. Melatonin analogs such as N-acetyl-serotonin and 5 hydroxytryptophol administered daily from 20-45 days of age did not produce any effect. Chronic melatonin administration from 20-50 days of life did not alter the occurrence of the nocturnal rise of circulating plasma melatonin, but did enhance its amplitude. Our results demonstrate that exogenous melatonin can inhibit or delay sexual maturation in the male rat if administered between 20 and 40 days of age and suggest that this inhibitory action is exerted at the hypothalamic and/or pituitary level. PMID- 6299702 TI - Effects of high doses of vitamin D3 and 1,25-dihydroxyvitamin D3 in lactating rats on milk composition and calcium homeostasis of the suckling pups. AB - Changes in serum Ca and phosphorus and in kidney Ca were determined in lactating rats and their suckling pups after the mothers received high doses of vitamin D3 or 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3]. High dietary vitamin D3 intake (300 IU/g diet) or daily oral doses of vitamin D3 (1 microgram/g BW) to vitamin I) replete (+D) lactating rats for 8 or 12 days caused significant increases in serum Ca in the mothers (1-2 mg/dl) and in their suckling pups (1.5 mg/dl). Daily oral doses of 1,25-(OH)2D3 (2 ng/g BW) to +D lactating rats caused a similar increase in serum Ca in the mothers, but did not affect the serum Ca of the pups. The administration of a high dose of 1,25-(OH)2D3 to vitamin D-deficient lactating rats or high doses of vitamin D3 to +D rats, caused no change in milk Ca, Mg, or phosphorus. Milk from +D rats given high doses of [3H]vitamin D3 (1 microgram/g BW) contained mostly [3H]vitamin D3 (85%) and a small amount of [3H]25-hydroxyvitamin D3 (6%). The results indicate that high doses of vitamin D3, but not 1,25-(OH)2D3, given to +D lactating rats can cause hypercalcemia in the suckling pups. The hypercalcemic effect on the pups observed after vitamin D3 treatment of the mother is probably a result of transport of toxic amounts of primarily vitamin D3 into the milk and is not due to altered mineral composition of the milk. PMID- 6299704 TI - Preparation of isolated bovine adipocytes: validation of use for studies characterizing insulin sensitivity and binding. AB - The present study was undertaken to develop techniques to isolate bovine adipocytes, to compare their rates of glucose metabolism and insulin sensitivity with adipocytes in adipose tissue, and to determine if isolated bovine adipocytes specifically bind insulin. Cell size and diameter distributions were the same for adipocytes fixed with OsO4 after isolation with collagenase and adipocytes liberated from OsO4-fixed adipose tissue slices. On a per cell basis, lipogenic rates were greater for isolated adipocytes compared with intact adipose tissue. Similar differences were found for glucose oxidation. In short term incubations, glucose oxidation and lipid synthesis were not stimulated by insulin (0-100 ng/ml) in either isolated adipocytes or tissue. Specific binding of [125I]iodoinsulin at 30 C was low (0.8%) in the first group of six beef cattle sampled, but increased with increasing cell concentration. Insulin degradation after 90 min was less than 5%. The specificity of [125I]iodoinsulin binding was studied in a second group of six animals. There was no specific binding of insulin in this group. In summary, bovine adipocytes can be isolated which are metabolically active and provide a valid system for studying hormone binding and action. In the present study, glucose metabolism in bovine adipocytes was not stimulated by insulin in vitro. This insensitivity to insulin was associated with a negligible capacity for insulin binding. These findings suggest that the lack of insulin sensitivity in bovine adipose tissue may be due to an inability to specifically bind insulin. This may be related to the unique metabolism of ruminant adipose tissue, which is less dependent upon glucose for fatty acid synthesis than is adipose tissue from nonruminant species. PMID- 6299703 TI - Mechanism of the direct action of gonadotropin-releasing hormone and its antagonist on androgen biosynthesis by cultured rat testicular cells. AB - The direct effects of GnRH and its agonistic and antagonistic analogs upon testicular androgen biosynthesis were studied in primary cultures of testicular cells obtained from adult hypophysectomized rats. Treatment of cultured cells with hCG (10 ng/ml) substantially increased testosterone production, while concomitant addition of GnRH or its agonist [des-Gly10, D-Ser(TBu)6,Pro9NHEt GnRH] decreased hCG-stimulated testosterone production in a dose-related manner with ED50 values of 1.2 X 10(-9) and 4.5 X 10(-11) M, respectively. Treatment with 10(-6) M of either a GnRH partial peptide or a cyclic GnRH analog did not affect hCG action; however, the addition of a GnRH antagonist ([Ac-D-Phe1,D-p-Cl Phe2,D-Trp3,6]GnRH) together with hCG and GnRH blocked the GnRH-induced decrease in testosterone production, with a half-maximal inhibitory dose ratio (antagonist to GnRH) of 0.15. The stimulatory effect of hCG became apparent by 8 h of incubation; no hCG effect was seen at this time in the presence of GnRH. Treatment with hCG increased cAMP accumulation, but GnRH administration did not affect hCG-induced cAMP accumulation. In contrast, treatment with GnRH depressed testosterone production induced by cholera toxin or (Bu)2cAMP. The inhibitory effect of GnRH on testosterone production (93% inhibition) was associated with decreases in hCG-induced 17 alpha-hydroxyprogesterone (39%) and delta 4 androstenedione (82%), but was not accompanied by a decrease in progesterone production. In cells incubated with cyanoketone and spironolactone to prevent pregnenolone metabolism, hCG stimulated pregnenolone biosynthesis, while concomitant GnRH treatment did not affect hCG action. In contrast, GnRH decreased hCG-induced testosterone production in cells treated with 10(-5) M progesterone. Similarly, GnRH decreased hCG-induced testosterone and androstenedione production in cells incubated with 10(-5) M 17 alpha-hydroxyprogesterone. The present results demonstrate that GnRH and its analogs exert direct actions on testicular cells through stereospecific recognition sites. The inhibitory effect of GnRH on testicular androgen production occurs at sites distal to the formation of cAMP and pregnenolone and may be due to decreases in the activity of the enzymes 17 alpha-hydroxylase and 17-20 desmolase. PMID- 6299705 TI - Luteinizing hormone receptors and testosterone production in whole testes and purified Leydig cells from the mouse: differences among inbred strains. AB - Testicular and Leydig cell function were studied in four inbred strains of mice. Significant strain-related differences were found in the number of LH receptors and in the production of testosterone in vitro in response to increasing concentrations of human CG (hCG) by both decapsulated testes and isolated Leydig cells. Maximal testicular testosterone production was similar in the C57BL/10J and C57BL/6J strains and considerably less in the DBA/2J (DBA) and C3H/HeJ strains, which were similar. However, the number of testicular LH receptors was less in C57BL/6J mice than in all three other strains. The pattern of maximal testosterone production by isolated Leydig cells among the four strains was similar to that observed for whole testes, whereas the number of LH receptors per 10(6) Leydig cells was least for Leydig cells from the DBA strain. When the number of Leydig cells per testis was estimated by dividing the number of testicular LH receptors by the number of LH receptors in 10(6) Leydig cells, it was apparent that testes from DBA mice contain approximately twice as many Leydig cells as those from the other strains. Differences in maximal testicular testosterone production appear to be reflected in differences in maximal testosterone production by isolated Leydig cells from each strain. However, there were no differences in testicular sensitivity to hCG but there were differences in Leydig cell sensitivity to hCG among the four strains. Furthermore, the existence of strain-related differences in well defined functional characteristics of Leydig cells offers the opportunity to study the genetic as well as physiological mechanisms involved in the regulation of Leydig cell function in normal individuals. PMID- 6299706 TI - 5,5'-Diphenylhydantoin decreases specific 3,5,3'-triiodothyronine (T3) binding by rat hepatic nuclear T3 receptors. AB - The influence of 5,5'-diphenylhydantoin (DPH) on specific T3 binding by rat hepatic nuclear T3 receptors was determined in tissue obtained from euthyroid and athyreotic rats. Nuclear binding of [125I]T3 was determined in isolated liver nuclei 60 min after the injection of DPH (20 mg/100 g BW) or vehicle (V). Mean specific nuclear T3 binding was decreased 14% (P less than 0.01) and 24% (P less than 0.02) from the control value in two experiments with nuclei from euthyroid rats and 23% (P less than 0.02) in nuclei from athyreotic rats. These findings in athyreotic rats and the observation that endogenous nuclear T3 was not increased in euthyroid rats after DPH injection suggested that the observed decrease in specific T3 binding after DPH injection did not result from displacement of endogenous T3 from plasma proteins or other binding sites into the nucleus. Further studies employing isolated nuclei revealed that the DPH-induced decrease in specific nuclear T3 binding appeared independent of duration of incubation up to 90 min and dose related. Maximum inhibition of specific T3 binding was 14.0% (P less than 0.001) and occurred in incubates that contained 320 microM DPH. Moreover, this DPH effect was evident throughout the full range of receptor saturation by T3. Similar effects of DPH were observed in studies using solubilized nuclear T3 receptors. A mean 14.0% decrement (P less than 0.001) in specific T3 binding was observed throughout the 72-h time course of these studies. Moreover, the DPH-induced changes occurred when solubilized receptors were up to 62% saturated with T3. These studies show that DPH injection or addition in vitro results in a 10-20% decrease in specific T3 binding by isolated hepatic nuclei or solubilized hepatic nuclear T3 receptors. This effect of DPH may result from a relatively weak interaction at nuclear sites and the very high concentration of DPH compared to T3 that prevails in DPH-treated animals. PMID- 6299707 TI - Binding, degradation, and utilization of plasma high density and low density lipoproteins for progesterone production in cultured rat luteal cells. AB - These studies were intended to examine the binding and degradation of plasma lipoprotein fractions and the utilization of lipoprotein-bound cholesterol for progesterone production in cultured rat luteal cells. These cells bound [125I]human low density lipoprotein ([125I]iodo-hLDL),[125I]human high density lipoprotein ([125I]iodo-hHDL), and [125I]iodorat HDL ([125I] iodo-rHDL) with high affinity. The equilibrium dissociation constants of the binding of labeled rHDL, hHDL, and hLDL were 90.5, 78.3, and 36.8 micrograms/ml, respectively. All three lipoproteins were also degraded in a concentration-dependent manner, with apparent Km values of 18.3, 17.5, and 22.4 micrograms/ml for rHDL, hHDL, and hLDL, respectively. The degradation of the lipoproteins was inhibited by lysosomotropic agents, transglutaminase inhibitors, and metabolic inhibitors, suggesting that these lipoproteins undergo internalization and lysosomal degradation. In addition, all three lipoproteins also augmented the hCG stimulated steroidogenesis. When cells were incubated with reconstituted LDL (the cholesterol ester in the LDL was replaced with [3H]cholesteryl linoleate) and the steroids produced identified, incorporation of tritium label in the progesterone fraction was observed in a time- and concentration-dependent manner. The incorporation of tritium into progesterone was increased by hCG and inhibited by an excess of unlabeled LDL in the incubation medium. These results show that the ovarian cells use lipoproteins as a source of cholesterol for steroidogenesis through receptor-mediated uptake and internalization, and the evidence suggests that the lipoproteins are intracellularly degraded. PMID- 6299708 TI - Characterization of binding and uptake of 3,3',5-triido-L-thyronine in cultured mouse fibroblasts. AB - The binding and internalization of 3,3'-[125I] 5-triiodo-L-thyronine ([125I]T3) was studied in cultured Swiss 3T3-4 mouse fibroblasts. At 0 C, the binding of T3 to cells is saturable, reversible, and stereospecific. These results together with those of earlier fluorescence studies using rhodamine-labeled T3 demonstrate the presence of specific plasma membrane T3 receptors. At 37 C, the uptake of T3 reached a steady state after 1 h, and approximately 57 fmol T3 were specifically taken up by 10(6) cells. In other cell lines, 7, 19, and 201 fmol T3 were specifically taken up by Chinese hamster ovary cells (subclone 10001), Kirsten sarcoma virus-transformed NIH 3T3 mouse fibroblasts, and nontransformed NIH 3T3 mouse fibroblasts, respectively. Incorporation of T3 into nuclei followed similar kinetics and accounted for approximately 9% of the total cellular uptake. Equilibrium binding studies of T3 to isolated nuclei showed one class of binding sites with an apparent association constant of 5 X 10(9) M-1 and a binding capacity of 16 fmol/100 micrograms DNA. At 37 C, the internalization of T3 was nearly totally blocked by antimycin A or rotenone, inhibitors of oxidative phosphorylation. These results indicate that the uptake of T3 is an energy dependent process. In the presence of bacitracin or monodansylcadaverine, substances that inhibit the receptor-mediated endocytosis of alpha 2 macroglobulin, the cellular uptake of T3 as well as the nuclear incorporation of T3 were inhibited in a concentration-dependent manner. The half-maximal inhibitory concentrations for the cellular uptake of T3 were 90 and 660 microM for monodansylcadaverine and bacitracin, respectively; for nuclear incorporation, they were 70 and 350 microM for monodansylcadaverine and bacitracin, respectively. These results indicate that receptor-mediated endocytotic uptake of T3 is a physiologically significant pathway. PMID- 6299709 TI - Cortisol inhibits and adrenocorticotropin has no effect on luteinizing hormone releasing hormone-induced release of luteinizing hormone from bovine pituitary cells in vitro. AB - Suckling causes a delay in onset of estrus and ovulation in cattle postpartum. In addition, the suckling stimulus causes the release of corticoids presumably via ACTH. Since any hormone released by suckling is a potential inhibitor of gonadotropin secretion and/or ovulation, we investigated the effects of ACTH and cortisol on LHRH-induced LH release from bovine pituitary cells in vitro. Anterior pituitary glands were obtained from cows killed during the luteal phase of an estrus cycle (days 5-15). Pituitary cells, disaggregated enzymatically, were grown in Dulbecco's medium containing 10% dextran charcoal-stripped fetal calf serum. On day 5, cultures were washed and reincubated in serum free medium containing the hormone being tested. After 6 h of incubation, LHRH was added in 10 microliters medium and incubation continued for an additional 6 h. ACTH at 4.3 X 10(-9), 4.3 X 10(-8), and 4.3 X 10(-7) M had no effect on basal or LHRH-induced LH release. Cortisol at 12.1 ng/ml decreased (P less than 0.001) the slope of LHRH response curve (b1 = 2.9 vs. 5.5 for controls). To determine if this effect was specific for cortisol, we compared cortisol, dexamethasone, and progesterone. LHRH-induced LH release (percent of control) was decreased (P less than 0.001) by 12.1 ng/ml cortisol (98%), 1, 5, and 10 ng/ml dexamethasone (60%, 71%, and 88%), but not by 3.1 ng/ml progesterone. The inhibitory effect of cortisol was reversible. Thus, LHRH-induced LH release (percent of controls) at 0, 24, 48, and 72 h after a 24-h exposure to 12.1 ng/ml cortisol was 19%, 89%, 100%, and 115%, respectively. We have demonstrated that cortisol at concentrations found normally in blood of cows postpartum will inhibit LHRH-induced LH release from bovine pituitary cells. This observation is consistent with the hypothesis that cortisol released by suckling may inhibit gonadotropin secretion postpartum and as such may prolong the anovulatory interval postpartum. PMID- 6299710 TI - Adenosine 3',5'-monophosphate (cAMP) and calcium-calmodulin interrelation in the control of prolactin secretion: evidence for dopamine inhibition of cAMP accumulation and prolactin release after calcium mobilization. AB - Calcium (Ca2+) ionophore A23187 increased the intracellular cAMP content and PRL release in normal rat anterior pituitary cells. Cotreatment with dopamine reduced both control and A23187-stimulated cAMP accumulation and PRL release. The dopamine antagonist spiperone restored the response of cAMP to ionophoric stimulation after pretreatment with dopamine in the greatest concentration used. Penfluridol, a compound with Ca2+-calmodulin-blocking properties, decreased control and A23187-stimulated cAMP content and PRL release. W7, a selective calmodulin-blocking agent, reduced basal cAMP and PRL release, whereas W5, a W7 analog with only 20% of its calmodulin-blocking ability, did not affect cAMP or PRL secretion. These data indicate that the Ca2+-calmodulin and cAMP systems are interrelated in the regulation of PRL secretion. They are also consistent with the hypothesis that the inhibition of PRL release by dopamine occurs after Ca2+ is mobilized and when or before it stimulates adenylate cyclase activity. PMID- 6299711 TI - Characterization of proopiocortin converting activity in rat anterior pituitary secretory granules. AB - Lysates from purified secretory granules of rat anterior pituitary glands were incubated with [3H]phenylalanine or [3H]arginine-labeled toad proopiocortin. The processed products formed were identified by immunoprecipitation with ACTH and beta-endorphin antisera, and by comigration with known markers on acid-urea polyacrylamide gels. Proopiocortin was cleaved by the secretory granule lysate primarily to 21,000 mol wt ACTH, 13,000 mol wt ACTH, 16,000 mol wt NH2-terminal glycopeptide, beta-lipotropin, a beta-endorphin-like peptide, and beta-endorphin. Characterization of the anterior pituitary proopicortin-converting activity shows that it: (1) cleaves specifically at the peptide bond on the carboxy side of the lysine-arginine residues of proopiocortin, (2) has a pH optimum in the acidic range, (3) is present in membrane and soluble fractions of the granule lysate, and (4) is inhibited by leupeptin, pepstatin A, and 2,2' dithiodipyridine, but not by p-chloromercuribenzoate, diisopropyl fluorophosphate, N alpha-p-tosyl-L lysine chloromethyl ketone hydrochloride, chloroquine, L-1-tosylamide-2 phenylethyl-chloromethyl ketone, or EDTA. PMID- 6299712 TI - Interactions of a monoclonal antibody to the insulin receptor with receptors for insulin-like growth factors. AB - A monoclonal antibody to the human insulin receptor was tested for its ability to inhibit the binding of 125I-insulin-like growth factor I (IGF-I) and 125I-insulin like growth factor II (IGF-II) to their receptors in human placenta membranes and cultured human IM-9 lymphocytes. In both placenta membranes and IM-9 cells, the antibody progressively inhibited the binding of 125I-IGF-I to its receptor with a potency that was 300-fold less than its ability to inhibit the binding of 125I insulin to its own receptor. In contrast, in human placenta membranes, this antibody inhibited the binding of 125I-IGF-II to its receptor only slightly. These studies indicate, therefore, that this monoclonal antibody binds preferentially to the insulin receptor but also crossreacts to a lesser extent with the IGF-I receptor. PMID- 6299713 TI - Differential effect of protein synthesis inhibition on TSH desensitization at different stages of primary thyroid cell culture. AB - In contrast to our previous experience with cultured thyroid cells, cycloheximide, actinomycin D and nicotinamide did not prevent TSH-induced desensitization in dog thyroid cells in primary culture for only one day. With continued duration of culture prevention of TSH desensitization by these agents did emerge, but asynchronously. Thus on the second day of primary culture, while cycloheximide and actinomycin D prevented TSH desensitization, nicotinamide remained ineffective. On the third day of primary culture all three agents blocked TSH desensitization. Examination of precursor incorporation into newly synthesized DNA, RNA and protein revealed a temporal association between the appearance of susceptibility to inhibition of TSH desensitization and an increase in DNA and protein synthesis. These data provide an explanation for the discrepant reports regarding the effect of protein synthesis inhibitors on TSH desensitization. PMID- 6299714 TI - Dynamics of cortisol and endorphin responses to graded doses of synthetic ovine CRF in sheep. AB - Corticotropin-releasing factor (CRF), recently isolated from sheep hypothalami, has been shown to stimulate secretion of ACTH and beta-endorphin in vitro, and in vivo in rat and man. In previous reports, responses to ovine CRF were studied in heterologous bioassay systems where the ovine sequence was likely to act as a CRF analogue. We administered synthetic ovine CRF to sheep to assess the dynamics of endorphin and cortisol responses. Graded doses of CRF caused a rapid increase in immunoreactive beta-endorphin (iB-E) within 2 min of iv administration, followed by a cortisol response which was maximal 15 min after the iB-E peak. Doses of CRF in excess of 10 micrograms did not increase the magnitude of the peak iB-E response but did prolong the duration of the plasma beta-endorphin rise. Ovine CRF is an extremely potent and rapidly-acting hypothalamic peptide in vivo when assayed in a homologous system. PMID- 6299715 TI - Effect of estradiol benzoate on hormonal profile and steroidogenic organs of immature and adult male rats. AB - The effects of estradiol benzoate (E2B) at a dose of 50 micrograms/day/rat for 15 days were investigated on ascorbate metabolism, steroidogenesis, protein, cholesterol levels of testis, adrenal and serum testosterone, LH and FSH profiles of rats. The data revealed that E2B manifested a direct effect on testicular and probably adrenal steroidogenesis. But serum gonadotrophin levels remained unchanged. The treatment also brought about a decline in ascorbate metabolism, activities of 3 beta and 17 beta hydroxysteroid dehydrogenases and alteration in protein level concomitant with the accumulation of cholesterol in both steroidogenic organs. Estrogen treatment was more effective in adult male rats than in the immature ones. PMID- 6299716 TI - Experimental observations on the mechanism of hormonal imprinting: influence of actinomycin D, methylamine and colchicine on receptor memory in a unicellular model system. AB - The first interaction between target cell and hormone gives rise to hormonal imprinting, which accounts for greater responsiveness of the cell at later interactions. The mechanism of hormonal imprinting is obscure; we based experimental approach to its closer study on combined treatment of Tetrahymena, as model cells, with diiodotyrosine (T2), which stimulates the division, and cell growth inhibitors, which interfere with different stages of cell reproduction, and methylamine, which inhibits cluster formation in the membrane. Of these, actinomycin D and methylamine inhibited the growth of the Tetrahymena, while colchicine did not, and all three suppressed the division stimulating action of T2, but could not prevent hormonal imprinting, as demonstrated on later re exposure to T2 of cells preexposed and not preexposed to T2 in combination with the inhibitors. It appears that the underlying mechanism of hormonal imprinting is highly complex, and involves many subcellular mechanisms and structures, but suppression of, or gross interference with, one or another of these cannot delete, only quantitatively reduce, the consequence of the first interaction with the hormone, i.e. hormonal imprinting. PMID- 6299717 TI - Role of cytoskeletal organization in the regulation of adenylate cyclase-cyclic adenosine monophosphate by hormones. AB - Microtubules, microfilaments, and intermediate filaments were found to be associated with the cytoplasmic face of the plasma membrane and even localized on the cell surface following "perturbation" of the plasma membrane. Several hormones interacting with their surface receptors have an effect on the assembly, organization, and orientation of the cytoskeletal system thus inducing changes in cell morphology, motility and aggregation. The cytoskeletal system is probably responsible for the lateral and vertical mobility of plasma membrane receptors and for the efficient coupling of GTP-binding protein to the adenylate cyclase moiety. It is suggested that the cytoskeletal system may be involved in hormone induced desensitization. The activity of cyclic nucleotide phosphodiesterase and protein kinase is modulated by Ca2+-calmodulin. These enzymes are associated with intermediate filaments and with microtubules which may control their activity and induce nuclear translocation of protein kinase. Stimulation of steroidogenesis by ACTH and LH, enhancement of H2O transport by vasopressin, elevation of the rate of amino acid and glucose transport by insulin, release of pancreatic insulin by glucose, and pituitary hormones by their respective hypothalamic releasing hormones, are only examples of a variety of hormonal responses that may be regulated by the cytoskeletal system. It is obvious that much more experimental study should be done to establish the role of the cytoskeletal system in hormonal action. I do hope this review will stimulate further ideas and experiments which might eventually lead to a better understanding of the role of the cytoskeletal system in the control of adenylate cyclase-cAMP system stimulated by hormones. PMID- 6299718 TI - The hormonal and local regulation of bone formation. PMID- 6299719 TI - A prospective study of infantile spasms: clinical and therapeutic correlations. PMID- 6299720 TI - Infantile spasms: some new theoretical aspects. AB - Infantile spasms begin in a very active phase of brain development. The new neuropathological techniques have shed light on changes in the fine structure of brain in children with infantile spasms. An understanding of the action of adrenocorticotropic hormone on the biochemistry and development of the brain provides, in an indirect way, a partial explanation of the biochemical events in this syndrome; most of these actions are well documented in animal experiments. Some anticonvulsants are also effective in treating infantile spasms, and modern research has explained many actions of these agents as well. The principal mechanism causing retardation in most children is unknown. Some future aspects of the research are reviewed. PMID- 6299721 TI - The hepatocyte primary culture/DNA repair assay using mouse or hamster hepatocytes. AB - The hepatocyte primary culture (HPC)/DNA repair assay using rat hepatocytes was developed to identify genotoxic chemicals. Since there are species differences in susceptibility to chemical carcinogens, it was desirable to extend the assay to other species. Carcinogens and noncarcinogens from six structural classes were tested with hepatocytes from B6C3F1 mice or Syrian hamsters. In hepatocytes from both species, DNA repair was elicited by the carcinogens methyl methanesulfonate, aflatoxin B1, 2-acetylaminofluorene, benzo(a)pyrene, dimethylnitrosamine, nitrosopyrrolidine, 3'-methyl-4-dimethylaminoazobenzene, and p dimethylaminoazobenzene. With aflatoxin B1, mouse hepatocytes required a dose of 10(-4) M for a maximum response while only 10(-6) was needed for hamster hepatocytes. All the presumed noncarcinogens, except 4-dimethylaminoazobenzene-4' sulfonyl chloride, were negative in mouse hepatocytes. This chemical, as well as aflatoxin G2 and pyrene, which have not been tested for carcinogenicity in the hamster, were positive in hamster hepatocytes. These findings demonstrate that genotoxic chemicals can be identified by the HPC/DNA repair assay using hamster or mouse hepatocytes. Furthermore, in vivo differences in susceptibility to chemical carcinogens such as aflatoxin B1 are reflected in the in vitro assay. PMID- 6299722 TI - Induction of ouabain-resistant mutants by chemical carcinogens in rat prostate epithelial cells. AB - We determined optimal conditions to quantitatively select ouabain-resistant (Ouar) mutants induced by chemical carcinogens in a rat prostate epithelial cell line (RPYK). These conditions included selection of Ouar mutants in 3 mM ouabain, an expression time of two days following a two-day treatment with carcinogens, and a reseeding density of 2 X 10(5) mutagenized cells per 100 mm dish to select mutants in ouabain. Ouar mutants induced by N-methyl-N'-nitro-N'-nitrosoguanidine (MNNG) remained stably Ouar when passaged in nonselective medium. Hemicyst formation, a characteristic of epithelial cells, was reversibly inhibited by ouabain in wild-type cells and was resistant to ouabain in Ouar cells. The direct acting carcinogens MNNG and methylazoxymethanol-acetate (MAMA) and the environmentally widespread procarcinogens aflatoxin B1 and benzo(a)pyrene increased the frequency of Ouar mutants in RPYK cells. The procedures developed here now make it possible to detect some environmental carcinogens likely to cause prostate cancer by virtue of their ability to mutate cultured prostate epithelial RPYK cells. The sensitivity of the RPYK cell line to aflatoxin-induced cytotoxicity and mutagenesis also makes it a useful cell system in which to study enzymes governing the conversion of aflatoxin to genotoxic metabolites. PMID- 6299723 TI - Health implications of natural fibrous zeolites for the Intermountain West. PMID- 6299724 TI - Cytotoxicity of a short-fiber chrysotile asbestos for human alveolar macrophages: preliminary observations. AB - Studies were performed to compare the cytotoxicity for human alveolar macrophages of a naturally occurring short-fiber chrysotile asbestos (RG 144) to that of a standard reference mixed-fiber (long and short) chrysotile asbestos (UICC chrysotile A. Rhodesian). Parallel studies were also performed with quartz (Min-U Sil 15), a known macrophage toxin. On a mass basis, and after 24 hr incubation, RG 144 was more cytotoxic than the UICC standard reference fiber and less toxic than quartz (silica). The cytotoxic potential of RG 144 chrysotile was further enhanced after size reduction by milling. These findings may have important biologic implications with respect to the use of short-fiber asbestos in industry. PMID- 6299725 TI - Effect of chrysotile and crocidolite on the morphology and growth of rat pleural mesothelial cells. AB - The effects of the UICC asbestos fibers chrysotile A and crocidolite on the morphology and growth of rat pleural mesothelial cells were examined. The response was different according to the fiber used. Within 4 hr of treatment with 5, 10, 20, or 50 micrograms/ml of chrysotile fibers, cell morphology showed intense vacuolization, in both logarithmic and confluent cells. Following treatment of growing cells with 20 or 50 micrograms/ml of chrysotile fibers, cell spreading was also observed. Four hours after treatment of cells with 5, 10, or 20 micrograms/ml of crocidolite fibers, no such changes were seen. Vacuolization appeared later and was much less marked than for chrysotile fibers. The mesothelial cell population-doubling time of about 30 hr was not significantly altered by addition of 5 to 20 micrograms/ml of crocidolite fibers, but a concentration of 50 micrograms/ml lengthened doubling time to about 90 hr. Treatment with 5 or 10 micrograms/ml chrysotile fibers usually prolonged this time, but 20 or 50 micrograms/ml caused cell lysis. After 48 hr of incubation with both types of fiber, the proportion of mitosis was the same as in control cultures, whatever the fiber concentration, and asbestos fibers were often seen inside dividing cells. PMID- 6299726 TI - Asbestos-induced alterations of human lymphoid cell mitogenic responses. AB - Using mitogenic assays, we have investigated the short term effects of two asbestos (amosite and chrysotile) fibers on lymphocyte functions in vitro. These oppositely charged fibers produced different alterations in mitogenesis. The blastogenic responses of concanavalin-A (Con-A) and pokeweed mitogen stimulated human peripheral blood mononuclear cells (PBMN) were significantly increased by the inclusion of 6 micrograms of chrysotile to the culture media. Amosite fibers proved to be inhibitory in all tests. When PBMN were depleted of monocytes, asbestos-related alterations of Con-A responsiveness were unchanged among the remaining cells. However, the addition of chrysotile to phytohemagglutinin (PHA) cultures resulted in a significant increase of the mitogenic response. When PBMN were enriched for T lymphocytes, and again cultured with the mitogens and fibers, the Con-A response still displayed impressive enhancement with chrysotile. In contrast to an intact PBMN population, PHA-induced blastogenesis among these T enriched lymphocytes was significantly elevated. These experiments demonstrate that asbestos can induce significant changes in the functional integrity of PBMN following a relatively short exposure time in culture. PMID- 6299727 TI - Wollastonite exposure and lung fibrosis. AB - Wollastonite is a naturally occurring acicular or fibrous metasilicate used in ceramics and as a substitute for asbestos in some applications. Wollastonite fibers are rather similar in form, length, and diameter to amphibole asbestos fibers but mineralogically they are different. Dust measurements in both the Finnish limestone--wollastonite quarry and in the flotation plant yielded high concentrations of both total dust and respirable fibers in some operational stages. The clinical study comprised a total of 46 men who had been exposed to wollastonite at the quarry for at least 10 years. Three of the fifteen nonsmokers showed chronic bronchitis. Radiographs revealed slight lung fibrosis among fourteen men, and slight bilateral pleural thickening among thirteen men. Their sputum specimens were normal. Spirometry and nitrogen single breath tests indicated the possibility of small airways disease. PMID- 6299728 TI - Evidence for a restriction/modification-like system in Anacystis nidulans infected by cyanophage AS-1. AB - Anacystis nidulans infected by AS-1 cyanophage contains an endonuclease (AS-1 endonuclease) which splits host DNA but not AS-1 phage DNA [Szekeres, M. (1981) Virology, 111, 1-10]. AS-1 phage DNA proved to be resistant not only to AS-1 endonuclease but also to a number of restriction endonucleases the recognition sites of which contain a central dG-dC dinucleotide. Since an unmodified 5'dG-dC dinucleotide was shown to be present at the sites at which DNA is cleaved by AS-1 endonuclease, the results suggest that the sites attacked preferentially by the AS-1 endonuclease are specifically protected on the AS-1 DNA molecule. The modification of AS-1 DNA was shown to occur specifically in infected Anacystis because AS-1 DNA fragments which are normally resistant to AS-1 endonuclease became susceptible to this enzyme if inserted into pBR322 plasmid and cloned in Escherichia coli. AS-1 DNA was shown to contain about 5% of a modified nucleotide which was not 5-methyldeoxycytidylic acid. Results presented and our earlier data suggest that in Anacystis infected by AS-1 phage, a restriction/modification-like system operates which is able to eliminate 'unwanted' (host) DNA selectively. PMID- 6299729 TI - Interaction of dicyclohexylcarbodiimide with the proton-conducting pathway of mitochondrial H+-ATPase. PMID- 6299730 TI - Faithful transcription of ribosomal 5-S RNA in vitro depends on the presence of several factors. AB - Cytoplasmic extracts from HeLa cells, capable of transcribing the cloned genes for ribosomal 5-S RNA, were employed to study the factors involved in this process. Two factors can be isolated, by gel filtration through Sephadex G-100, which are devoid of RNA polymerase activity. They significantly enhance the extent and specificity of the transcription of 5-S rRNA. Both proteins can jointly be purified by affinity chromatography on immobilized DNA containing the genes for ribosomal 5-S RNA from Xenopus borealis. Besides a protein of approximately 45 kDa, possibly corresponding to TF IIIA isolated from Xenopus oocytes, a second protein with a molecular mass of 22 +/- 1 kDa stimulates the formation of 5-S RNA. This protein is contained in the breakthrough of DEAE cellulose; it binds to and is eluted from phosphocellulose with 0.6 M KCl. In addition, it was found that the exclusion volume obtained after gel filtration on Sephadex G-100 contains functional complexes, which are capable of transcribing the cloned 5-S genes and hence contain all the required factors. Direct evidence is presented that the protein of 22 kDa described above is contained in and can be isolated from such complexes. It is postulated from indirect evidence that an additional factor with a molecular mass in excess of 100 kDa is required which can be removed from functional polymerase complexes by gel filtration through Bio Gel A5m. PMID- 6299731 TI - Yeast mutant with thermolabile CDP-choline synthesis. Isolation and characterization of a cholinephosphate cytidyltransferase mutant. PMID- 6299732 TI - A calcium-activated protease which preferentially degrades the 160-kDa component of the neurofilament triplet. AB - A calcium-dependent protease fully active with 0.2 mM Ca2+ was found associated with the neurofilament-enriched cytoskeleton of the rat spinal cord prepared by the treatment with Triton X-100. The enzyme preferentially degrades the 160-kDa component of the neurofilament triplet. In addition, a soluble calcium-dependent protease activity was found in the supernatant from the spinal cord, which degraded a variety of cytoskeletal proteins including the neurofilament triplet, glial fibrillary acidic (GFA) protein, actin, tubulin, and a high molecular weight protein associated with microtubules. The possibility that the cytoskeleton-bound activity is an artefactual association of the soluble enzyme to the cytoskeleton seems to be negated on the basis of the following dissociation and reassociation experiments. The protease activity remained associated with the cytoskeleton in the physiological ionic strength, and was not completely dissociated from it until the KC1 concentration was raised to 0.6 M. When the 0.6 M KCl-extract was dialysed against salt-free buffer to remove KC1, and added back to the protease-free cytoskeletal pellet, proteolytic activity was partially restored. Full activity returned only when the extract and the protease free cytoskeletal pellet were first combined in the presence of 0.6 M KC1, and then slowly reassociated by dialysis against salt-free buffer. Dissociated enzyme was rapidly inactivated at 37 degrees C in the presence of Ca2+. These results suggest the structural association of the protease with the cytoskeleton under the physiological condition. PMID- 6299733 TI - Mitochondrial pantetheinephosphate adenylyltransferase and dephospho-CoA kinase. PMID- 6299734 TI - Structural and functional properties of cytochrome c oxidase from Bacillus subtilis W23. AB - The terminal component of the electron transport chain, cytochrome c oxidase (ferrocytochrome c: oxygen oxidoreductase) was purified from Bacillus subtilis W23. The enzyme was solubilized with alkyglucosides and purified to homogeneity by cytochrome c affinity chromatography. The enzyme showed absorption maxima at 414 nm and 598 nm in the oxidized form and at 443 nm and 601 nm in the reduced form. Upon reaction with carbon monoxide of the reduced purified enzyme the absorption maxima shifted to 431 nm and 598 nm. Sodium dodecylsulfate polyacrylamide gel electrophoresis indicated that the purified enzyme is composed out of three subunits with apparent molecular weights of 57 000, 37 000 and 21 000. This is the first report on a bacterial aa3-type oxidase containing three subunits. The functional properties of the enzyme are comparable with those of the other bacterial cytochrome c oxidases. The reaction catalyzed by this oxidase was strongly inhibited by cyanide, azide and monovalent salts. Furthermore a strong dependence of cytochrome c oxidase activity on negatively charged phospholipids was observed. Crossed immunoelectrophoresis experiments strongly indicated a transmembranal localization of cytochrome c oxidase. PMID- 6299735 TI - The use of affinity chromatography on 2'5' ADP-sepharose reveals a requirement for NADPH, thioredoxin and thioredoxin reductase for the maintenance of high protein synthesis activity in rabbit reticulocyte lysates. PMID- 6299736 TI - Primary structure of the DNA-binding protein HRm from Rhizobium meliloti. AB - The amino acid sequence of protein HRm, a DNA-binding HU-type protein of 90 residues (Mr 9303), isolated from Rhizobium meliloti, has been established from automated sequence analysis of the protein and from structural data provided by peptides derived from cleavage of the protein at arginine and aspartic acid residues. The comparison of the primary structure of protein HRm with that of other HU-type proteins shows that two short sequences, of 7 and 6 residues respectively, located in the median part of the molecule, appear highly conserved and may be important in the function of the protein. PMID- 6299737 TI - Interaction between isolated cytochrome c1 and cytochrome c. AB - Cytochrome c1 from bovine heart mitochondria was isolated by a modification of the technique of Konig et al. [(1980) Biochim. Biophys. Acta 621, 283-295] which involved an affinity chromatography step on a gel with yeast cytochrome c as a ligand. Its spectra, electrophoretic pattern in presence of sodium dodecylsulfate, its reducibility by ascorbate and cytochrome c were characteristic of a native cytochrome, with a single polypeptide having an apparent molecular weight of 30 000. By using an arylazido derivative of cytochrome c, having the photoactive group bound to lysine 13, upon illumination a cross-link with the described preparation of cytochrome c1 was obtained. By pepsin digestion of the cross-linked complex a limiting fragment was obtained and partially sequenced. It allowed to identify the site of binding of cytochrome c near the sequence 167-174 of cytochrome c1. PMID- 6299738 TI - Proteolytic conversion of angiotensins in rat brain tissue. AB - The proteolytic conversion of angiotensins in rab brain preparations was studied. Angiotensin I was converted into angiotensin II by enzymes which were associated with a synaptic membrane preparation, while angiotensin II was relatively resistant to proteolysis by these enzymes. Angiotensin II was rapidly metabolized at both pH 7.4 and pH 5.4 by enzymes in the soluble fraction of a synaptosomal preparation. One of the fragments formed at pH 7.4 was characterized as angiotensin III. At pH 5.4 only one fragment was generated which was characterized as angiotensin-(1-7)-heptapeptide. Enzymatically generated angiotensin II and III displayed pronounced biological activity in the brain, whereas angiotensin-(1-7)-heptapeptide was inactive. These data indicate a route for the generation, and the inactivation of biologically active angiotensins in the brain. PMID- 6299739 TI - Regulation of UDPG pyrophosphorylase in Acetabularia mediterranea. AB - The kinetic properties of UDPG pyrophosphorylase (glucosyl-1-phosphate uridyl transferase, EC 2.7.7.9) suggest that it may play a key role in the regulation of metabolism in Acetabularia mediterranea. The enzyme-catalyzed reaction is readily reversible in vitro, and has been assayed in both directions. The enzyme shows substrate inhibition by UDPG and UTP at substrate concentrations in excess of 2 mM. The kinetic behavior of the enzyme is consistent with the hypothesis that it catalyzes an ordered bisubstrate biproduct reaction in which G-1-P is the leading substrate, and UTP is the leading product. A plot of initial velocity vs. PPi concentration is sigmoid, indicating a cooperative homotropic effect. PGAL inhibits the reaction in the direction: UTP + G-1-P leads to UDPG + PPi It has no effect on the reverse reaction. The responses of the enzyme may serve to regulate the allocation of G-1-P between anabolic and catabolic pathways. PMID- 6299740 TI - Ultrastructure of the purified and reconstituted Na/K-ATPase of the avian salt gland. AB - Na/K-ATPase of salt-stressed salt glands of the domestic duck (Anas platyrhynchos) was purified in membrane-bound form by incubation of the microsomal fraction with sodium dodecylsulphate and ATP followed by discontinuous sucrose gradient centrifugation. Gel electrophoresis of the purified plasma membrane preparation substantially showed the two polypeptide subunits of the Na/K-ATPase both of which stained with the periodic acid-Schiff reagent. About 99% of the total ATPase activity was ouabain-inhibitable amounting to 1300 mumol Pi/(mg protein X h) of specific activity. The anion-stimulated, ouabain insensitive ATPase increased parallel to the Na/K-ATPase up to the microsomal fraction until it totally vanished during SDS incubation. Electron microscopy of thin sections revealed that the purified fraction consisted of flat and cup shaped triple-layered membrane fragments. Particles arranged into clusters and strands were visible as 3 to 5 nm surface particles in negatively stained suspensions and as 8 to 10 nm intramembraneous particles in freeze fracture replicas. The differential distribution of the intramembraneous particles on the fracture faces reflected the structural membrane asymmetry. Solubilization of Na/K-ATPase led to the disappearance of intramembraneous particles. Incorporation of the solubilized enzyme into phosphatidylcholine vesicles again showed 8 to 10 nm particles apparently orientated at random in the artificial membrane. Control liposomes prepared in the absence of solubilized enzyme were devoid of intramembraneous particles. These results clearly demonstrate that the avian salt gland Na/K-ATPase exists as 8 to 10 nm particles in both the purified plasma membrane and the artificial phospholipid membrane. PMID- 6299741 TI - Right ventricular myocardial infarction diagnosed by 99 m technetium pyrophosphate scintigraphy: clinical course and follow-up. AB - Out of 178 consecutive patients with acute inferior wall myocardial infarction submitted to technetium-99 m pyrophosphate scintigraphy, 49 (27.5%) were found to have concomitant right ventricular infarction. Gated blood pool scans showed right ventricular abnormalities in 21 out of 26 patients who were submitted to this investigation (right ventricular asynergy: 16 cases; right ventricular dilatation: eight cases; decreased right ventricular ejection fraction: 16 cases). Complications were common in the acute stage. Shock was noted in 19 cases (eight related to bradycardia, three related to relative hypovolaemia and eight instances of true cardiogenic shock). Atrial fibrillation (seven patients), ventricular fibrillation (eight patients) and severe atrioventricular conduction disorders (13 patients) were also frequent. In spite of this, the in-hospital mortality was low: three deaths occurred (6.1%), one from heart failure, two others from posterior septal rupture. All patients were followed up for one year or more. Six additional deaths were noted (three from left cardiac failure, two from recurrent anterior wall infarction and one from massive pulmonary embolism). Clinical assessment, haemodynamic measurements and gated blood pool scans showed significant improvement of right ventricular function with return to normal in those cases with small right ventricular infarcts as judged from technetium-99 m pyrophosphate scintigraphy. In spite of the complications seen in the initial period, patients with a right ventricular infarction have a good overall prognosis and the long-term outcome, primarily determined by the left-sided lesions, is often favourable. PMID- 6299742 TI - An immuno-enzymatic assay for herpes simplex virus tumour associated antigen in gynecological oncology. AB - The authors studied the HSV-TAA (Herpes Simplex Virus Tumor Associated Antigen) in patients affected by female genitale tract tumors, using the immunoenzymatic assay (ELISA). They found a positive frequence of 65% in sera of patients affected by uterine cervical carcinoma and of the 80% in sera of patients affected by vulvar carcinoma. The authors suggest that this enzymatic method has a real value and propose its use in the early diagnosis of the female genital tract neoplasms. PMID- 6299743 TI - Cervical mullerian adenosarcoma: case report. PMID- 6299744 TI - 99mTc-methylene diphosphonate uptake in a primary Wilms' tumor. PMID- 6299745 TI - Emergency blood pool scintigraphy in fresh-blood-aspirated focal liver disease. AB - Emergency blood pool scintigraphy was performed in a patient clinically diagnosed as having a liver abscess, in whom percutaneous drainage revealed that the lesion containing fresh blood. 99mTc-RBC images 18 h after the injection showed only patchy activity in the lesion, therefore, we considered there was no communication between the lesion and the circulating blood. Drainage of the lesion was performed again and the abscess with a blood clot was drained. PMID- 6299747 TI - Counting the costs of cancer therapy. AB - The costs of cancer treatment were estimated for five groups of cancer patients: those with operable breast cancer receiving adjuvant chemotherapy and patients with advanced breast cancer, ovarian cancer, germ cell cancer and small cell lung cancer. Cancer treatment costs were substantial but chemotherapy cost was a relatively minor item. Methods of limiting treatment costs are identified as reducing hospital stay, encouraging outpatient administration of chemotherapy and limiting the number of supernumary investigations. The cost of different cancer treatment options should be considered when management decisions are made. PMID- 6299746 TI - Binding of 99mTc-gluconate to heart mitochondria. AB - 99mTc-gluconate has previously been shown to bind to isolated rat heart mitochondria. In the presence of potassium cyanide this binding is enhanced several fold, an effect which has been shown to be correlated to the efficiency of the in vivo binding of the isotope complex in ischemic dog hearts. The present investigation shows that the potassium cyanide-induced binding was mainly localized to the mitochondrial inner membrane; in the presence of cyanide other organelles, e.g., nuclei and liver microsomes also showed some binding. Boiling, lipid extraction, or addition of N-ethylmaleimide caused a variable inhibition of the binding of 99mTc-gluconate to rat heart mitochondria, which also was markedly influenced by temperature, pH, time, and concentrations of protein and isotope complex, but not by conditions affecting energy levels or calcium transport. Fractionation of submitochondrial particles exposed to 99mTc-gluconate in the presence of potassium cyanide indicated that the 99mTc-gluconate-binding component cofractionated with cytochrome oxidase. It is postulated that a protein component localized in the mitochondrial inner membrane, possibly cytochrome oxidase, is responsible for the binding of 99mTc-gluconate. PMID- 6299748 TI - Modified ribonucleosides as biological markers for patients with small cell carcinoma of the lung. AB - A variety of individual modified ribonucleosides may be elevated in the urine of cancer patients. They can be readily measured quantitatively in a single reversed phase high-performance liquid chromatographic run. A total of 41 patients with small cell carcinoma of the lung were studied. For 5-ribonucleosides determined in the pretreatment urine of 28 patients, the respective frequency of elevation was directly related to stage of disease. One or more nucleosides were evaluated in the pretreatment urine of 27 out of 28 patients (96%). Included were 11 patients with limited disease and 10 (91%) had 2 or less than 2 nucleosides elevated, whereas 16 out of 17 (94%) with extensive disease had 3 or more elevated. Based on this same discriminant, median survival was significantly extended for patients with 2 or less nucleosides elevated (24 months) in contrast to 3 or more (10 months). Using a single number to represent the summation of equally weighted individual nucleoside values as a composite score, a direct relationship was found between increasing extent of disease or tumor burden. This was in contrast to more variable results for carcinoembryonic antigen analyzed in plasma samples obtained at the same time. When determined serially the composite score paralleled in general the clinical response categories for individual patients. PMID- 6299749 TI - Demonstration of immunoreactive calcitonin in sera and tissues of lung cancer patients. AB - In sera of 194 patients with lung carcinoma, calcitonin was estimated by radioimmunoassay with an antibody against human calcitonin. Increased levels of calcitonin-immunoreactive protein were found in 57% of the patients with small cell carcinoma, in 10% of the patients with squamous carcinoma and in only 2 patients with large cell carcinoma. In patients with small cell carcinoma, serial determinations of calcitonin were accomplished during therapy. Significantly decreased calcitonin levels were found in patients who responded to therapy with cytostatics and X-ray. Increased calcitonin levels were measured from 1 to 2 months before clinical symptoms of a relapse were detectable. Investigations on the biochemical nature of this calcitonin-immunoreactive protein were made on both serum of lung cancer patients and tumor tissue. Evidence for the production of calcitonin-immunoreactive protein directly by the tumor was given by immune histology and by determinations of calcitonin in tumor tissue. Three protein fractions, which were immunoreactive with anti-human calcitonin, with molecular weights of about 100,000, 48,000 and 20,000, were separated by gel filtration. The two higher molecular weight fractions were degraded to molecular weights of about 17,000 and about 3400--the molecular weight of physiological calcitonin--by incubation with sodium dodecyl sulphate under reducing conditions. These results led to the conclusion that it may be possible to characterize a tumor-specific calcitonin precursor molecule; in addition to its use in monitoring therapy it may be useful in the differential diagnosis of small cell carcinoma. PMID- 6299750 TI - A viral antigen as a marker for the prognosis of human breast cancer. AB - An antigen present in human breast tumor cells, and which is immunologically related to the envelope protein (gp52) of murine mammary tumor virus, was used as a marker for the detection of breast cancer in an Israeli population. The results show that the antigen was detectable in 128 of 204 breast carcinomas tested (62.7%). The immunological reaction was not detected in normal breast tissue, benign breast tumors, ductal hyperplasia or in primary malignancies in other organs. A significantly higher percentage of cases with demonstrable antigen was found in Israeli women born in North Africa (78%) as compared to women of European origin (60.6%). The frequency of detection of the antigen was higher in stage IV (80%) as compared to stage I (15%), suggesting that the gp52 cross reacting antigen is a marker for the severity of the disease. Moreover, a retrospective study of 97 cases of stage II breast cancer shows that if the antigen is detected at the time of mastectomy, one can usually predict an unfavorable prognosis. Survival data analysis indicates that patients without detectable antigen survived significantly longer than those with a detectable antigen. PMID- 6299751 TI - Herpes virus-related antigens in herpes simplex virus type 2-transformed cells in the course of cervical carcinoma. AB - Anticomplement immunofluorescence (ACIF) technique was applied to Herpes simplex virus type 2 (HSV-2)-transformed cells (333-8-9) and to cells lytically infected with HSV-2 in sera from women with cervical carcinoma. There was a correlation between the positive results with both types of cells. Long-term survivors suffering from cervical cancer showed a high percentage of positive immunofluorescence (IF) reactions in HSV-2 transformed cells (77%) as well as in cells lytically infected with HSV-2 (77%) in sera obtained before treatment. The figures were 81 and 83% respectively in sera collected 6-24 months after treatment. When the IFs of survivors and non-survivors were compared, the survivors showed more cytoplasmic staining reactions in sera obtained before treatment than in similar sera from non-survivors, whereas there was no difference in nuclear IF. The sera of survivors taken 6-24 months after treatment had a significantly higher overall fluorescence rate than similar sera from non survivors. In late sera taken 36 months after treatment, a drop in reactivity was noted for the survivors to values comparable with those obtained from non survivors. The controls (age-matched, healthy women and patients with malignancies other than cervical carcinoma) showed significantly less IF reactions (P less than 0.001). PMID- 6299752 TI - Absence of an effect of mianserin on the actions of clonidine or methyldopa in hypertensive patients. AB - The concurrent administration of tricyclic antidepressants has been shown in man to result in a clinically significant impairment of the antihypertensive effect of clonidine. This interaction is thought to be related to competition for central alpha 2 receptors where clonidine acts as an agonist and the tricyclics act as antagonists. Although it seems to cause less cardiovascular effects than tricyclic antidepressants, the tetracyclic antidepressant, mianserin also has been reported to be an alpha receptor antagonist and may, therefore, also interfere with the antihypertensive activity of centrally-acting drugs. This study investigates the effects of acute and chronic mianserin administration in patients with essential hypertension established on long term treatment with either clonidine or methyldopa. The first dose of mianserin was not associated with an increase in blood pressure and during a further two weeks of mianserin therapy there were no significant alterations in blood pressure, supine or erect. Similarly, mianserin did not alter heart rate either after acute or after chronic administration. Mianserin itself had a sedative effect but there was no interference with the sedation attributable to clonidine or methyldopa. Mianserin caused no reduction salivary flow and did not influence the reduced saliva production caused by clonidine. Both clonidine and methyldopa are associated with a reduction in sympathetic outflow but there was no evidence in this study of any further change in plasma noradrenaline or 24 h urinary catecholamine excretion. This study demonstrates that if mianserin is given acutely or chronically, it does not interfere with the effects of the centrally acting anti-hypertensive drugs, clonidine and methyldopa. Mianserin may therefore be a suitable antidepressant for patients receiving these antihypertensive agents if drug treatment for depression is indicated. PMID- 6299753 TI - Biological activity of anti-thyrotropin anti-idiotypic antibody. AB - Rabbit anti-rat anti-human thyrotropin anti-idiotypic antibodies have been raised. These antibodies were active at the thyrotropin (TSH) receptor, in that they inhibited 125I-labeled bovine TSH binding to thyroid plasma membranes, stimulated adenylate cyclase activity through a guanyl nucleotide-dependent mechanism, augmented radioiodide transport into isolated porcine thyroid follicular cells and induced such cultured cells to organize into follicles. Aside from substantiating the expectation that anti-hormone anti-idiotypic antibodies may possess properties of the original hormone, this work raised the possibility that thyroid-stimulating antibodies which cause the hyperthyroidism of Graves' disease may be anti-TSH anti-idiotypic antibodies. PMID- 6299754 TI - Evidence for the ontogenic precedence of suppressor T cell functions in the human neonate. PMID- 6299755 TI - Solubilization and assay of [3H]imipramine binding sites from human platelets. AB - [3H]Imipramine binding sites were characterized on outdated (72-96 h) human platelets. The binding site profile was very similar to that previously described for fresh platelets and for CNS membrane preparations. [3H]Imipramine binding sites are believed to represent the recognition site for the 5-hydroxytryptamine uptake system. In order to further investigate this molecular relationship, we solubilized [3H]imipramine binding sites using 1% digitonin and characterised them using a polyethylene glycol precipitation assay. Solubilization did not alter the pharmacological specificity or affinity of the [3H]imipramine binding site and recovery of sites was very high. It was concluded that the solubilization procedure left the binding site remarkably intact. The ready availability of platelets and the convenience of the binding assay makes this system amenable for further purification steps. PMID- 6299756 TI - Autoradiographic localization of high-affinity [3H]kainic acid binding sites in the rat forebrain. AB - Utilizing in vitro autoradiographic techniques, we have studied the distribution of high affinity [3H]kainic acid ([3H]KA) binding sites in intact sections of the rat forebrain. These sites have the same kinetic and pharmacological characteristics as the [3H]KA site described in tissue homogenates. Moderate to high levels of specific binding were observed in several discrete brain regions. These include lamina I, V and VI of the neo- and cingulate cortex, superficial layers of the pyriform cortex, striatum, external plexiform and granule cell layers of the olfactory bulb, olfactory tubercle, the stratum lucidum of CA3 of the hippocampus, molecular layer of the dentate gyrus, reticular nucleus of the thalamus, the hypothalamic median eminence, and the granule cell layer of the cerebellum. Low levels of specific binding were associated with other discrete regions such as the lateral septum, bed nucleus of the stria terminalis, medial geniculate, superficial layers of the superior colliculus, nuclei of the central grey, interpeduncular nucleus and the molecular layer of the cerebellum. Moderate uniform levels of specific binding were observed over the hypothalamus, zona incerta and the amygdala. One of the important factors in KA neurotoxicity seems to be the presence of KA receptors, and regions that are susceptible to the toxic effects of KA after local administration, such as the striatum, hippocampus, amygdala and pyriform cortex, have moderate to high levels of binding. Thus, these data provide a useful map for studying the relationship between receptor mediated and seizure-induced neuronal damage following KA administration. PMID- 6299757 TI - Opiate binding sites in bovine retina: evidence for benzomorphan selective binding sites. PMID- 6299758 TI - Increase in potencies of opioid peptides after peptidase inhibition. AB - Various agents that have been reported to reduce the enzymatic degradation of the enkephalins have been tested for their ability to potentiate the activity of [Met5]enkephalin in three in vitro assay tissues. The greatest effect was obtained with the combination of bestatin (10 microM or 30 microM), captopril (10 microM), thiorphan (0.3 microM) and L-Leucyl-L-leucine (2 mM) which increased the potency of [Met5]enkephalin 18-fold in the guinea-pig myenteric plexus, 13-fold in the mouse vas deferens and 200-fold in the rat vas deferens. The increased potency is attributed to inhibition of the peptidases since the mixture of inhibitors did not change the activity of either normorphine or the metabolically stable synthetic opioid peptides. The potencies of the hexa-, hepta- and octapeptide C-terminus extensions of [Met5]enkephalin and [Leu5]enkephalin were increased by the peptidase inhibitors in all three preparations; the greatest effects were found in the rat vas deferens. No significant changes in the potencies of fragments of beta-endorphin longer than beta-endorphin-(1-19) were obtained. It may now be possible to inhibit enzymatic degradation of opioid peptides sufficiently to measure their release from neurones activated by electrical field stimulation. PMID- 6299759 TI - Augmentation of acetylcholine response in denervated skeletal muscle by endorphins and spinal cord-conditioned culture media. PMID- 6299760 TI - Lack of involvement of alpha 2-adrenoceptors in the regulation of striatal dopaminergic transmission. PMID- 6299761 TI - Comparison of photoaffinity labeling of P2-purinergic receptors of isolated guinea-pig vas deferens by arylazido aminopropionyl ATP and by arylazido aminobutyryl ATP. AB - Two chemically related arylazido photoaffinity analogs of ATP (arylazido aminopropionyl ATP (ANAPP3) and arylazido aminobutyryl ATP (ANABP3)), which have been reported in the literature to differ in their ability to inhibit myosin ATPase, were compared for their ability to antagonize contractile responses of the isolated guinea-pig vas deferens to ATP. During photolysis in organ chambers the photoconversion (delta A260/delta t) of ANAPP3 occurred with greater than first order kinetics or was multiexponential and t1/2 = 7.5 min, while delta A260/delta t for ANABP3 was first order and t1/2 = 2.25 min. After photolysis of these compounds in the presence of the guinea-pig vas deferens, using irradiation periods which caused 80% consumption of the compounds, ANABP3 was 2-3 times more potent than ANAPP3 in antagonizing contractions to ATP, which are mediated by P2 purinergic receptors. A comparison of concentration-response curves obtained for nonphotolyzed ANAPP3 and ANABP3 used as agonists suggested that the greater antagonism produced by photolyzed ANABP3 is not attributable to a greater potency. The results suggest that the longer 3'-hydroxyl-arylazide bridge length of ANABP3 places the arylnitrene intermediate in a position at or near the P2 receptor which is more favorable for covalent insertion. PMID- 6299762 TI - Differences in the applicability of the easson-stedman hypothesis to the alpha 1- and alpha 2-adrenergic effects of phenethylamines and imidazolines. AB - The enantiomers of 2-(3,4, alpha-trihydroxybenzyl)imidazoline and the corresponding desoxy derivative, 2-(3,4-dihydroxybenzyl)imidazoline, were evaluated at alpha 1- and alpha 2-adrenergic receptors to test the applicability of the Easson-Stedman hypothesis to the imidazoline class of alpha-adrenergic agonists. A series of closely related phenethylamines was included for comparison. The Easson-Stedman hypothesis states that optically active adrenergic agonists possessing an asymmetric hydroxyl-substituted benzylic carbon atom will have the following relative potencies: R(-) greater than S(+) = desoxy. While the phenethylamines were found to adhere to the Easson-Stedman hypothesis at both alpha 1- and alpha 2-adrenergic receptors, the optically active imidazolines did not. These findings further support our previous observations that the phenethylamines and imidazolines may interact differently with alpha-adrenergic receptors. PMID- 6299763 TI - Ageing as a factor governing the effect of isoproterenol on the cyclic AMP level in isolated aorta from spontaneously hypertensive and normotensive rats. PMID- 6299764 TI - Evidence that Substantia Nigra is crucial to neural network of kindled seizures. PMID- 6299766 TI - Reinitiation of host DNA synthesis in senescent human diploid cells by infection with Simian virus 40. AB - Human diploid fibroblasts, TIG-1, cease to proliferate at about 60-62 population doubling level. In their senescent state used in this study, the percentage of nuclei labeled by [3H]thymidine for 48 h was around 1-2% in fresh medium containing 5-40% fetal bovine serum. The percentage of labelled nuclei increased up to 10-fold after infection with SV40. This increase reflects stimulation of cell DNA synthesis because: 1. The increase also occurred when ts A900 was used for infection at the non-permissive temperature, under these conditions viral DNA synthesis is inhibited; 2, the increase paralleled the stimulation of [3H]thymidine incorporation into DNA in a Hirt-precipitate fraction from SV40 infected cells. UV-irradiated SV40 had reduced ability to induce DNA synthesis. A viable deletion mutant of SV40, d1940, had almost the same activity to induce cell DNA synthesis as did wild-type SV40. Equilibrium density gradient centrifugation analysis of DNA labelled with 5-bromodeoxyuridine (BrdU) supported semiconservative replication rather than repair synthesis. We conclude that a considerable fraction of human diploid cells in a senescent population initiate host DNA replication by infection with SV40, although these cells cannot be stimulated with fetal bovine serum. PMID- 6299765 TI - Dissociation of morphine's analgesic and respiratory depressant actions. PMID- 6299767 TI - Detection of avian hematopoietic cell surface antigens with monoclonal antibodies to myeloid cells. Their distribution on normal and leukemic cells of various lineages. AB - The isolation and characterization of monoclonal antibodies reacting with cell surface antigenic determinants of normal and leukemic avian hematopoietic cells is described. The antibodies were produced by immunizing mice with normal macrophages, as well as with myeloid cells transformed with the avian acute leukemia viruses MC29, AMV and E26. Eleven antibodies were characterized for their reactivity with a variety of normal and leukemic cells of the myeloid, B- and T-lymphoid and of the erythroid cell lineage. Using several methods, they could be subdivided into five distinct types: I. Four antibodies were specific for the myeloid lineage, predominantly reacting with immature myeloid cells. II. One antibody reacted with mature and immature myeloid cells as well as with T lymphoid cells. III. Four antibodies reacted with myeloid, erythroid and T lymphoid cells. IV. One antibody reacted with myeloid as well as with T- and B lymphoid cells. V. One antibody reacted with all kinds of chicken hematopoietic cells except erythrocytes. The first type of antibodies detected glycoproteins with MWs of 170 and 130 kD. The pattern of antigens precipitated varied with the different monoclonal antibodies of this group. The antibody of the fourth type precipitated a 30 kD polypeptide from extracts of myeloid and lymphoid cells. None of the other antibodies precipitated any detectable proteins. PMID- 6299768 TI - Fusion-mediated injection of SV40-DNA. Introduction of SV40-DNA into tissue culture cells by the use of DNA-loaded reconstituted Sendai virus envelopes. AB - Sendai virus envelopes can be solubilized by non-ionic detergents such as Triton X-100. Removal of the detergent from a supernatant containing the solubilized viral envelope glycoproteins results in the formation of reconstituted fusogenic viral envelopes. When SV40-DNA is added to the reconstitution system, it is trapped within the viral envelope. Incubation of SV40-DNA-loaded Sendai virus envelopes with permissive cells (CV1 and TC7 cells) resulted in fusion-mediated injection of the trapped DNA, as was demonstrated by the ability of the injected cells to synthesize SV40-T-antigen. Quantitative estimation revealed that up to 20% of the injected cells were able to synthesize T-antigen. Loaded viral envelopes were able to inject SV40-DNA and to promote synthesis of T-antigen also in cells which are resistant to infection by intact SV40 viruses, such as F1' 1-4 cells. In addition, it is shown that reconstituted envelopes of Sendai virus are able to transfer membrane fragments from SV40 receptor-positive into SV40 receptor-negative cells, such as F1' 1-4 cells. After implantation of SV40 receptors, the F1' 1-4 cells became susceptible to infection by intact SV40 viruses. PMID- 6299769 TI - A primary role for microfilaments, but not microtubules, in hormone-induced cytoplasmic retraction. AB - The distributions of microfilaments and microtubules were studied during transient hormone-induced changes in cell shape (retraction-respreading). Two cell types (fibroblasts and bone cells), differentially responsive to parathyroid hormone (PTH) and prostaglandin E2 (PGE2), were analysed. The cytoplasm of fibroblasts retracted in response to PGE2 but not PTH, whereas bone cells could respond to both PGE2 and PTH. Time-lapse photomicrography indicated that the retraction began within minutes of hormone addition, while respreading occurred over longer times, up to 8 h. Affinity-purified actin and tubulin antibodies were used to follow the appearance of microtubules and microfilaments during both the retraction and the respreading phases. Microtubules appeared not to reorganize noticeably, although they were squeezed closer together in cellular pseudopods; no extensive loss or growth was detectable. Microfilaments did alter drastically their appearance and distributions. Soon after hormone addition when earliest detectable cytoplasmic retraction was evident, microfilament bundles appeared to break down. Remaining microfilament bundles consisted of relatively short, non aligned fragments or aggregates. During respreading, microfilament bundles regrew and realigned throughout the cytoplasm. These data suggest a primary role for microfilaments, but probably not microtubules, in these cell shape changes. PMID- 6299770 TI - Three-drug chemotherapy combined with radiation therapy in small cell carcinoma of the lung. AB - In 43 cases of small cell carcinoma of the lung, a combined treatment has been initiated with three drugs (cyclophosphamide 750 mg/m2, adriamycin 50 mg/m2 and vincristine sulphate 1 or 2 mg total dosage), split-course-radiation therapy on the primary tumour (3500 rads) and prophylactic irradiation of the brain (2000 rads). The median survival of the 34 cases evaluable at day 50 attains 253 days. A more favourable evolution is observed for patients with a good response after therapy (median survival: 315 days) and for cases with limited disease (321 days) than for non-responders (median survival: 157 days) and for cases with extensive disease (median survival: 214 days). In spite of tumour site irradiation, prophylactic irradiation of CNS and chemotherapy, there were six local relapses, two CNS extensions and six metastatic relapses and only two autopsied cases without macroscopic evidence of relapse. PMID- 6299771 TI - Pulmonary histiocytomas. AB - Five cases of pulmonary histiocytomas are presented (four "endobronchial" and one "pulmonary"). They occurred in two children, and three adults over a five year period. Our center has a catchment of 1 million inhabitants. The possible nature and diagnostic possibilities are discussed. The prognosis is good in young people with this disease, which, in principle, does not differ from the same proliferating histiocytic and fibroblastic disorder arising elsewhere. The histological diagnosis was based upon characteristic and consistent morphological features, common to all cases. PMID- 6299772 TI - Neuropharmacological properties of electrophysiologically identified, visually responsive neurones of the posterior lateral suprasylvian area. A microiontophoretic study. AB - Extracellular recordings have been made from 118 electrophysiologically identified neurones lying in the posterior lateral suprasylvian area (PLLS and PLMS) of cats anaesthetized with Nembutal. Eighty-one cells were activated synaptically by the electrical stimulation of cortical and subcortical sites known to be the sources of monosynaptic projections to the lateral suprasylvian area; latencies to such activations have been measured. The locations and sizes of the receptive fields of 55 neurones were determined. The direction sensitivity and ocularity of these cells also were examined. The effects of various pharmacological agonists and antagonists have been observed on visual responsiveness and synaptic excitability. The excitatory effects of subcortical (dorsal lateral geniculate nucleus and pulvinar nuclear complex) electrical stimulation on the activity of suprasylvian neurones were reduced substantially by the iontophoretic administration of atropine. Antagonists of the receptors for the excitatory amino acids reduced the effectiveness, on the single cell evoked activity, of stimulation of the ipsilateral 17/18 border region and contralateral homotopic lateral suprasylvian area. Both classes of antagonist reduced the magnitude of neuronal responses to photic stimulation, and these response attenuations were additive when the antagonists were ejected concurrently. All of the pharmacological effects were reversible and reproducible. These data lend support to the proposition that acetylcholine and an excitatory amino acid are mediators of synaptic transmission of cortical visual processes in the lateral suprasylvian area. PMID- 6299773 TI - An electrophysiological characterization of projections from the pedunculopontine area to entopeduncular nucleus and globus pallidus in the cat. AB - Stimulation of the pedunculopontine region (PPN), in which neurons are filled by horseradish peroxidase injected into the entopeduncular nucleus (ENTO) or globus pallidus (GP) of the cat, excites ENTO and GP neurons both orthodromically and antidromically. Stimulus threshold mapping experiments and intracellular records of EPSPs provide evidence that the orthodromic excitation may be produced monosynaptically by the axons of PPN neurons. Antidromic excitation of ENTO axons from stimulation in PPN or the thalamus may elicit IPSPs in ENTO neurons via the action of recurrent collaterals. An excitatory synaptic action of PPN neurons on pallidal cells could be a partial basis for the high discharge rate characteristic of these neurons in awake animals. PMID- 6299774 TI - Intracellular study of presumed interneurons in the ventrobasal complex of cats with chronic cortical ablations. AB - In a ventrobasal neuronal population surviving chronic ablation of somatosensory and motor cortices, the input-output properties of cells activated by ML fibers and showing functional characteristics of interneurons (large peripheral receptive fields, convergence of different types of receptor afferents, repetitive discharge) were studied. Since intracellular recordings showed the presence of excitatory and inhibitory postsynaptic potentials, it is argued that inhibitory interneurons also receive inhibitory inputs, suggesting that data processing in VB is more complex than hitherto postulated. PMID- 6299775 TI - Physical characterization of the E. coli dnaC region carried by a plaque forming gamma dnaC transducing phage. PMID- 6299776 TI - The inhibitory amino acid GABA hyperpolarizes motor axons: an intracellular study. PMID- 6299777 TI - cGMP stimulates active K+ uptake in rat submandibular slices. AB - The 8-bromo derivative of cGMP was found to stimulate the ouabain-sensitive uptake of K+ and to reduce the net release of K+ induced by acetylcholine in rat submandibular gland slices incubated in vitro. PMID- 6299778 TI - Influence of chronic UV-light exposure on hepatic and cutaneous monooxygenases. AB - Hairless female Ng/-mice were irradiated by UV-light for 16 h daily over a period of 24 weeks. Monooxygenase activities were measured in liver and skin, and an induction of the aryl-hydrocarbon hydroxylase was detected in liver by both fluorometric and radiochemical methods, whereas no induction of this enzyme could be demonstrated in the skin. PMID- 6299779 TI - Separation of VX-2 rabbit carcinoma-derived cells capable of releasing collagenase. AB - Primary and secondary cultures of VX-2 carcinoma produced high levels of collagenase activity in both active and latent forms in serum-free media. These cultures appeared morphologically heterogeneous in phase-contrast microscopy and revealed the presence of mainly three distinct forms: epithelial-like cells (E cells), fibroblast-like cells (F cells), and large rounded-flat cells which may represent a subclass of the F cells. Cell separation techniques such as brief dispase treatment, Percoll gradient centrifugation, thimerosal treatment, and rabbit serum were used to obtain predominantly one form or the other. The E cells never formed a monolayer but rather grew as limited size clusters of intimately associated cells with large nuclei and often appeared multinucleated. These cells were difficult to maintain in culture or serially passed more than a few times. The F cells, rare in early cultures but having the highest growth potential, appeared in various morphological forms ranging from spindle- to stellate-shaped cells. The cells in their third passage were capable of producing palpable tumors, similar in light and electron microscopic studies to the original tumor from which they were derived, when injected intramuscularly into recipient rabbits and produced specific collagenase activity in active and latent forms in serum-free media. Ultrastructural studies suggested that the E cells were of epithelial origin whereas the F cells were similar to stromal fibroblasts. Cytogenetic studies demonstrated that almost all of the E cells showed both numerical and structural chromosomal changes in a modal number of 54 chromosomes. On the other hand, the major cell population of the F cells resembled normal rabbit fibroblasts; both contained a normal diploid (2n = 44). However, few cells (4-6%) in the F-cell population were hyperdiploid with a modal chromosome number of 54. These cells may represent inadvertent contaminating E cells and account for the apparent limited turmorigenicity observed in early F-cell cultures. The data suggested that the E cells were of tumor origin whereas the majority of the F-cell population appeared to be of host origin. Furthermore, it is suggested that the E cells stimulate tumor-associated stromal cells to produce elevated levels of collagenolytic activity and contribute to collagen degradation during tumor invasion. PMID- 6299780 TI - Cytotoxic action of quartz dust on stimulated and nonstimulated peritoneal macrophages in vitro. AB - Quartz dust cytotoxicity was studied in starch-stimulated and nonstimulated mouse peritoneal macrophage cultures in vitro. Morphological changes, release of lactate dehydrogenase (LDH) into the culture medium, and lactate production were used as parameters of cell damage. Phagocytosis of quartz dust by macrophages increased cell detachment of both the stimulated and nonstimulated cells. Both cell types released significant amounts of LDH into the supernatant culture medium. The increased LDH release was more from stimulated dust-free and corundum and quartz-exposed macrophages than from nonstimulated macrophages. There was no statistically significant difference in lactate production between stimulated and nonstimulated macrophage cultures in the presence or absence of quartz or corundum dust. PMID- 6299782 TI - Alternative hypotheses of proton ejection in cytochrome oxidase vesicles. Transmembrane proton pumping or redox-linked deprotonation of phospholipid cytochrome c complex(es). AB - A review of published experimental and interpretative knowledge concerning proton ejection associated with cytochrome c oxidation by artificial phospholipid vesicles inlaid with cytochrome c oxidase indicates that the detailed characteristics of the redox-linked proton ejection cannot be simply explained by proton pumping. We propose an alternative hypothesis according to which proton ejection is due to the redox-linked deprotonation of a complex involving phospholipid and cytochrome c at the surface of the vesicles. The postulates upon which this hypothesis depends are explicitly outlined, and some methods of testing the hypothesis are suggested. PMID- 6299781 TI - Ultrastructural responses by Golgi apparatus of rat submandibular gland to beta adrenergic, alpha-adrenergic, and cholinergic stimulation. AB - Rat submandibular gland tissue pieces were stimulated in vitro for 30 min with a beta-adrenergic agent or a cyclic AMP analog to stimulate protein secretion, or with alpha-adrenergic or cholinergic agents or a Ca2+ ionophore to stimulate fluid secretion. Acinar cells were examined by transmission electron microscopy. In control tissue, acinar cells showed little evidence of secretory activity. The Golgi apparatus was sparse and was associated with a few small, immature secretory granules with fine fibrillar contents. Following secretory granule discharge stimulated by isoproterenol or dibutyryl cyclic AMP, acinar cells were constricted, and had extensive basolateral membrane folding and tightly packed rough endoplasmic reticulum. Golgi complexes were prominent and had multiple small granules with filamentous contents. After stimulation of fluid secretion by alpha-adrenergic agents (epinephrine, phenylephrine), or cholinergic agents (acetylcholine, carbachol, pilocarpine), or a Ca2+ ionophore (A23187), the Golgi apparatus had compact concave cisternae enclosing aggregates of tubulovesicles. Acinar cells were distended, basolateral membranes were expanded, and rough endoplasmic reticulum was dilated and vesiculated. PMID- 6299783 TI - Compound 48/80-induced permeability change in liposomal membrane. AB - The action of compound 48/80 (a mixture of condensation products of p-methoxy-N methylphenethylamine with formaldehyde) on liposomal membranes was studied by means of K+-loaded liposomes and a K+ ion-selective electrode. Prompt efflux of K+ was detected when 48/80 was added to the negatively charged liposome suspension, while the monomer of 48/80, p-methoxy-N-methylphenethylamine, did not release K+ from the same liposomes. The mechanism for the action of 48/80 is discussed in comparison with that of a polymyxin, well known as an antibiotic acting on bacterial membranes. PMID- 6299784 TI - Electron spin resonance study on the mechanism of polyethylene glycol-membrane interaction. PMID- 6299785 TI - Induction of ornithine decarboxylase by treatment of guinea pig lymphocytes with phospholipase C. AB - Treatment of guinea pig lymphocytes with Clostridium perfringens phospholipase C but not with Naja naja snake venom phospholipase A2 increased ornithine decarboxylase activity. The increase in ornithine decarboxylase activity was suppressed by actinomycin D or cycloheximide, suggesting that de novo syntheses of RNA and protein are necessary for the increase in the enzyme activity. These results suggest that the activation of phospholipase C rather than that of phospholipase A2 is responsible for induction of ornithine decarboxylase during lymphocyte transformation. PMID- 6299786 TI - A new sequence-specific endonuclease from Gluconobacter suboxydans. PMID- 6299787 TI - Acylation: a new post-translational modification specific for plasma membrane associated simian virus 40 large T-antigen. AB - SV40 transformed mouse cells (mKSA) were labeled in parallel with either [35S]methionine or [3H]palmitate and subfractionated. Nuclear extracts and solubilized plasma membranes were analyzed for the presence of either 35S- or 3H labeled SV40 large tumor antigen by immunoprecipitation and SDS polyacrylamide gel electrophoresis. The majority of the [35S]methionine labeled large T was recovered from the nuclear fraction, only minor amounts were detected in plasma membranes. In contrast, large T labeled specifically with [3H]palmitate was found only in the plasma membrane fraction. Our results demonstrate a specific acylation of large T associated with plasma membranes, suggesting that the membrane location of this predominantly nuclear protein is specific. PMID- 6299788 TI - Molecular sizes of photolabeled GABA and benzodiazepine receptor proteins are identical. AB - [3H]Muscimol was irreversibly incorporated into rat cerebellar membranes upon irradiation with ultraviolet light. GABA agonists and antagonists inhibited this incorporation. The reversible muscimol-binding decreased after photoaffinity labeling of the membranes with muscimol. These results indicated that this irreversible incorporation is to the GABA receptor. Photolabeled GABA receptor protein showed its Mr of 50000 +/- 1000 in SDS-polyacrylamide gel electrophoresis. This molecular size is identical to that of the benzodiazepine receptor which was photolabeled with 3H-flunitrazepam. PMID- 6299789 TI - An overlapping site model for the lipid annulae of membrane proteins. PMID- 6299790 TI - A high yield method for the preparative synthesis of coenzyme A by combination of chemical and enzymic reactions. AB - Dried cells of Brevibacterium ammoniagenes are a good enzyme source for the preparative synthesis of CoA from pantothenic acid, L-cysteine and ATP. A problem with this synthesis is that the CoA synthesis is repressed by negative feedback inhibition by CoA to pantothenate kinase, the first step enzyme for the biosynthesis of CoA, which catalyses phosphorylation of pantothenic acid or pantetheine. As the inhibition operates only at this step, a further increased yield is possible if the enzymic phosphorylation step is replaced with chemical synthesis. Yields from phosphorylated substrates are more than 10-times higher than those from pantothenic acid or pantetheine (33 g/l from phosphopantothenic acid with a molar yield of 86%; 115 g/l from phosphopantetheine with a molar yield of 100%). PMID- 6299791 TI - Z-DNA and other non-B-DNA structures are reversed to B-DNA by interaction with netropsin. AB - The interaction between the B-form specific ligands netropsin (Nt) and distamycin 3 (Dst-3) and DNA duplexes has been studied under conditions of salt concentration and low water activity that modify the polymer conformation into a non-B DNA form, putatively a Z-like form. Three polymers with strict alternating purine-pyrimidine sequences and GC content from 100-0% have been tested: poly(dG dC) . poly(dG-dC), poly(dA-dC) . poly(dG-dT) and poly(dA-dT) . poly(dA-dT). The titrations by Nt and Dst-3 were followed by circular dichroism. Although specific binding of Nt to the Z-form of poly(dG-dC) . poly(dG-dC) does not occur, Nt reverses this Z structure to the B-type conformation; Dst-3 is, however, totally inefficient. The presumed non-B or Z-like structure of poly(dA-dC) . poly(dG-dT) is reversed to the B-form upon interaction with Nt; Dst-3 also induces this reversal but at higher ligand ratios. The modified B-structure of poly(dA-dT) . poly(dA-dT) in low water activity is efficiently reversed to the B-form by interaction with both Nt and Dst-3. PMID- 6299792 TI - The position of polypeptide G on the encephalomyocarditis virus polyprotein cleavage map. AB - The two-dimensional mapping of tryptic peptides of encephalomyocarditis virus specific proteins has demonstrated that the amino acid sequence of non-structural polypeptide G constitutes a portion of the molecule of a precursor of capsid proteins, polypeptide A. The results of pulse-chase in vitro translation experiments strongly suggest that polypeptide G corresponds to a C-terminal moiety of polypeptide A. Variations in the polyprotein cleavage maps of different picornaviruses are briefly discussed. PMID- 6299793 TI - Characterization of sulphydryl groups of the mitochondrial phosphate translocator by a maleimide spin label. AB - A maleimide spin label strongly inhibits the phosphate/H+ symporter of rat liver mitochondria. While inducing half-maximal inhibition of transport, the spin label reacts preferentially with the SH groups of the carrier, which are at least of two types. One type of SH group is localized close to the surface of the membrane and its environment does not significantly influence the mobility of the probe. The second type of SH group is buried in the membrane, is not accessible to ascorbate or chromium oxalate and its environment greatly restricts the motion of the probe. PMID- 6299794 TI - Effect of sodium butyrate on the stimulation of casein gene expression by prolactin. AB - Sodium butyrate, but not isobutyrate, inhibits prolactin action on the induction of casein synthesis and casein mRNA accumulation in rabbit mammary explants. Sodium butyrate specifically prevents the generation of the prolactin relay which can be released from isolated membranes incubated with prolactin and which stimulates directly casein gene transcription when added to isolated mammary nuclei. This indicates that sodium butyrate exerts its inhibitory action essentially at the membrane level. PMID- 6299795 TI - Metabolite-controlled phosphorylation of hepatic phosphofructokinase proceeds by cAMP-dependent protein kinase. AB - In hepatocytes 32P-incorporation into rat liver phosphofructokinase is stimulated by glucose as well as by glucagon, the effects of both stimuli being prevented by L-alanine [Eur. J. Biochem. (1982) 122, 175]. The phosphopeptides of the enzyme derived from limited proteolysis by subtilisin and from exhaustive tryptic digestion were analyzed either by one-dimensional mapping on sodium dodecyl sulphate-polyacrylamide slab gels and by fingerprint mapping, respectively. It is shown that in vivo stimulation of 32P-incorporation by glucose or by glucose plus glucagon results in identical phosphopeptide maps, and that these maps were identical with those obtained from phosphofructokinase phosphorylated in vitro with catalytic subunit of cAMP-dependent protein kinase. It is concluded that in the intact liver cell phosphofructokinase is phosphorylated by cAMP-dependent protein kinase but that the state of phosphorylation is modified by metabolite control. PMID- 6299796 TI - Cellular localization of the receptor-dependent and receptor-independent uptake of human LDL in the liver of normal and 17 alpha-ethinyl estradiol-treated rats. AB - The cellular localization in the liver of the receptor-dependent and -independent uptake of human low density lipoprotein (LDL) in normal and 17 alpha-ethinyl estradiol-treated rats was investigated by the simultaneous in vivo injection of human 131I-LDL and human reductive methylated 125I-LDL. The cells were subsequently isolated by a low temperature method. In untreated rats, after 30 min of in vivo circulation of human LDL, 57% of the receptor-dependent liver association of human LDL occurs in non-parenchymal cells and 43% in parenchymal cells. Estradiol treatment of rats for 3 days selectively increases the receptor dependent cell-association of human LDL with hepatocytes (17-fold), while the receptor-dependent cell-association with non-parenchymal cells is not affected. PMID- 6299797 TI - Characterization of CHAPS-solubilized prolactin receptors induced by estradiol in liver of male rats. Unmasking of cryptic sites by CHAPS. PMID- 6299798 TI - Guanine nucleotides can activate the insulin-stimulated phosphodiesterase in liver plasma membranes. AB - The insulin-stimulated cyclic AMP phosphodiesterase from liver plasma membranes is shown to be activated upon incubation with guanine nucleotides in the presence of ATP. The non-hydrolysable analogue of ATP, adenylyl imidodiphosphate failed to substitute for ATP in achieving activation. GTP, its non-hydrolysable analogues p[NH]ppG and GTP-gamma-S, as well as GDP, all elicited activation. It is suggested that guanine nucleotides, and probably insulin, exert their effect on this enzyme through a distinct species of guanine nucleotide regulatory protein. PMID- 6299799 TI - The presence of LHRH-like receptors in Dunning R3327H prostate tumors. AB - Quantitative analyses of LH-RH-like membrane receptors were performed in five tumors from the transplantable Dunning R3372H rat prostatic adenocarcinoma. The binding of D-Trp6-LH-RH, an agonist of LH-RH, was observed in all 5 tumors. The antagonist [Ac-Dp-Cl-Phe1,2,D-Trp3,D-Lys6, D-Ala10]-LH-RH was bound to 4 tumors. The apparent equilibrium dissociation constant (Kd) for D-Trp6-LH-RH receptor was from 2.6-3.9 x 10(-10) M. The apparent equilibrium Bmax values (maximum number of binding sites) were from 17.2-86.0 fmol/mg membrane protein for D-Trp6-LH-RH receptor. The Kd for the antagonist was from 2.4-2.7 x 10(-10) M and the Bmax values were from 35.5-66.0 fmol/mg membrane protein. Similar binding studies performed in 6 normal rat prostates showed no binding capacities. PMID- 6299800 TI - An evaluation of the evidence for H+ pumping by reconstituted cytochrome c oxidase in the light of recent criticism. AB - We review the evidence for H+ translocation by reconstituted cytochrome c oxidase; attention is paid to those aspects which we feel most open to criticism. Possible alternative hypotheses are assessed, with regard to experiments carried out to test them directly and with regard to published data. We conclude that, whilst certain aspects of this system are worthy of further clarification, the reported observations are all consistent with proton translocation by reconstituted cytochrome c oxidase; most of these observations provide a positive indication of such an activity. PMID- 6299801 TI - The turnover of vimentin in Ehrlich ascites tumour cells. AB - The turnover of vimentin and vimentin-derived peptides has been examined in logarithmically growing Ehrlich ascites tumour cells. Cells were pulse-labelled with [35S]methionine for 30 min and then chased for up to 60 h. It was found that the specific radioactivity of the main isoelectric variant of vimentin decreased to half the original value in 15.3 h which was close to the division time of the cells (16 h). The protein moiety of the phosphorylated variant of vimentin also turned over very slowly, in contrast to the turnover rate of the phosphate group itself which has a half-life of 1.4 h. The role of the intermediate filament specific, Ca2+-activated proteinase has been considered in relationship to the slow turnover of vimentin. PMID- 6299802 TI - Evidence that amiloride antagonises insulin-stimulated protein phosphorylation by inhibiting protein kinase activity. AB - The diuretic drug amiloride antagonises the insulin-dependent increase in phosphorylation of ATP-citrate lyase in hepatocytes isolated from rats that had been starved and refed a fat-free diet. Studies with a range of protein kinases and protein phosphatases that have been shown to phosphorylate or dephosphorylate purified ATP-citrate lyase in vitro revealed that amiloride was a non-specific inhibitor of all protein kinases tested, but did not significantly affect any of the protein phosphatases. These results cast doubt on previous claims that growth factors stimulate phosphorylation of ribosomal protein S6 by activating an amiloride-sensitive Na+/H+ exchange system, and that insulin inhibits a protein phosphatase that is activated by amiloride. PMID- 6299803 TI - Structure subtraction as an approach to investigation of the mechanism of restriction enzyme action. AB - Endonuclease BamHI cleaves the phosphodiester bonds between the guanine residues within the duplex DNA sequence G decreases GATCC. The substrate characteristics of oligonucleotides, containing some defects in the sequence recognized by endonuclease (nick, absence of some internucleotide phosphate or nucleotide, partially single-stranded form of the recognition site) were investigated. The results suggest that the specificity of synthetic oligonucleotide cleavage is strongly dependent on the ribosophosphate backbone intactness inside the recognition site. BamHI was found not to hydrolyse the phosphodiester bonds outside the double helix. Also BamHI forms a productive complex with the non symmetrical substrate, having half the recognition sites, of a single strand. PMID- 6299804 TI - Electrochemically driven respiration in mitochondria and Paracoccus denitrificans. The coupling of the electrochemistry of horse heart cytochrome c with respiration in mitochondria and a model thereof, Paracoccus denitrificans. AB - By exploiting the rapid, direct electrochemistry of horse heart cytochrome c at a modified gold electrode it has been possible to couple the electrode reaction with respiration in rat liver mitochondria and in protoplasts of Paracoccus denitrificans, but not in protoplasts from E. coli. Oxidation of endogenous and exogenous sources of reducing equivalents via cytochrome c is also observed. PMID- 6299805 TI - Effect of transmethylation-reaction and increased levels of cAMP on superoxide generation of guinea-pig macrophages induced with wheat germ agglutinin and phorbor myristate. AB - Superoxide (O2-) generation of guinea-pig macrophages induced by wheat germ agglutinin (WGA) was suppressed to a great extent by the inhibition of transmethylation with 3'-deazaadenosine. When macrophages were stimulated with phorbor myristate (PMA) instead of WGA, the suppression of O2- generation of macrophages was observed to be slight despite the presence of 3'-deazaadenosine. These results were confirmed under various conditions. Thus the WGA-stimulated O2 generation of macrophages is probably associated with transmethylation, but the PMA-stimulated O2- generation is not. WGA-stimulated O2- generation of macrophages was also inhibited in the presence of dibutyryl cAMP or prostaglandin E2 (PGE2), substances that increase intracellular cAMP, but PMA-stimulated O2- generation was only slightly affected by these compounds. These results suggest that the mechanism for O2- generation of macrophages caused by WGA is different from that for O2- generation caused by PMA. PMID- 6299806 TI - Functional development of the cervical sympathetic pathway in the neonatal rat. AB - Maturation of the cervical sympathetic innervation of the levator palpebrae smooth muscle was studied physiologically in vivo in the rat. Tonic pre- and postganglionic nerve activity and responses to centrally acting stimuli, asphyxia and hypoglycemia, were recorded. Development of functional capacity of the sympathetic nerve terminal-smooth muscle complex was assessed by measuring contractions evoked by directly acting noradrenergic agonists, endogenous nerve terminal norepinephrine released by tyramine, and electrical stimulation of preganglionic axons. Tonic preganglionic activity and responses to asphyxia and hypoglycemia were fully developed by the second postnatal day. Ganglionic neurotransmission, however, is not established until the end of the first postnatal week so that sympathetic outflow from the central nervous system (CNS) does not affect the nerve terminal-smooth muscle complex before this time. Functional capacity of the complex develops in a stepwise fashion with intervening plateau periods. There is an initial rapid increase before onset of ganglionic neurotransmission, and development to the mature level is attenuated only modestly by decentralization of the ganglion on the first postnatal day. It is concluded that conduction of impulses to the postganglionic neuron is not essential to initiate maturation of this nerve terminal-smooth muscle complex, that a significant degree of maturation occurs before ganglionic transmission is established, and that maturation to the adult level is usually largely independent of neural connections between the CNS and the postganglionic neuron. PMID- 6299807 TI - Maturation of circulatory responses to adrenergic stimuli. AB - The postnatal maturation of renal and femoral circulatory responses to catecholamines and to stimulation of their efferent sympathetic nerve supply has been examined in developing swine. Catecholamine dose-response experiments were carried out in intact animals under pentobarbital anesthesia. Effects of denervation and nerve stimulation were studied in intact animals and effects of neurotransmitter infusions were studied in preparations for in situ perfusion of kidney or hind limb. The renal circulation was found to be relatively sensitive to norepinephrine and under tonic neural vasoconstrictor influence at birth, and the femoral circulation was not. The femoral circulation was found to be relatively sensitive to isoproterenol at birth, whereas the renal circulation exhibited beta-adrenergic vasodilation by the end of the first postnatal week. The basic pressure-flow relationship in either circulation at any age was not altered by infusions of neurotransmitters into the arterial circuit after denervation of the perfused kidney or hind limb. Adrenergic innervation continued to mature rapidly in both circulations during the first postnatal week, as evidenced by the decreasing threshold and increasing magnitude of vasoconstrictor responses to electrical stimulation of the renal or lumbar nerves. A cholinergic component of the response to lumbar nerve stimulation became functional at 1 month after birth. PMID- 6299808 TI - An overview of current information on bioavailability of dietary iron to humans. AB - Bioavailability factors can greatly modify the absorption of dietary iron consumed in different meals by an individual. A greater percentage is generally absorbed of heme iron from animal tissues than of nonheme iron of either animal or plant food. The amount of meat in a meal is the only bioavailability factor known to influence absorption of heme iron. Absorption of iron from the exchangeable nonheme iron pool of a meal is influenced by both enhancing and inhibiting substances or factors. Ascorbic acid, meat, fish, and poultry enhance absorption of nonheme iron, and meals may be classified according to relative bioavailability depending on the content of meat, fish, poultry, and/or ascorbic acid. Some low-molecular-weight organic acids may also increase the bioavailability of nonheme iron. Synthetic metal-chelating agents added to foods and the beverages tea and coffee will inhibit absorption of nonheme iron in a concentration-dependent manner. Wheat bran, soy products, cow's milk, and egg tend to decrease bioavailability of nonheme iron when included in a meal. However, the effect of compounds thought to be responsible for the inhibition in purified form (phytate, fiber, phosphoproteins) is dependent on chemical form and concentration. In some foods there may be as yet unidentified inhibitors or interaction between compounds to inhibit absorption of nonheme iron. Currently available information permits estimation of relative bioavailable iron in a meal. PMID- 6299809 TI - [Effect of bradykinin and angiotensin on synaptic transmission in smooth muscles]. PMID- 6299810 TI - [Histamine action on the ionic currents in the smooth muscle cells of guinea pig ureter]. PMID- 6299811 TI - [Protective properties of nicotinamide and cytochrome C in aminoglycoside ototoxicosis]. PMID- 6299812 TI - [A simple method for radioimmunoassay of total estrogen in urine]. AB - A simple and rapid radioimmunoassay (RIA) without extraction or purification was developed for Total Estrogen (conjugated and unconjugated estrogen) in urine from normal men and nonpregnant women. Antiserum used to RIA was produced by immunizing rabbits with estriol-16 alpha-glucuronide (E3-16-G)-BSA conjugates. Antiserum to E3-16-G.BSA significantly cross-reacted with E1 (100%), E2 (100%), E3 (100%), E3-16-G (100%), AND E3-17-G (100%) and did not react with other conjugated estrogen in urine. Urinary estrogen glucuronides and sulfates were gently hydrolyzed by beta-glucuronidase from Helix pomatia juice for 2hr (48 degrees C) without inhibition, and hydrolysis urine was directly applied to RIA. The results obtained from this direct method correlated well with the chromatographic purificating method, the extracting method and the Brawn Kambegawa method (colorimetric method). This direct method is very useful for clinical application. PMID- 6299813 TI - [The age-related changes in serum 17-hydroxyprogesterone secretion in men]. AB - In order to clarify the age-related changes in serum 17-hydroxyprogesterone (17 OHP) secretion in men, serum basal 17-OHP levels were determined in 203 healthy male subjects from 10 months to 116 years of age, and serum 17-OHP responses to dexamethasone (Dex), fluoxymesterone (FM), HCG and ACTH were compared between the young and elderly groups. The mean basal 17-OHP level remarkably increased from pubescence to the third decade and gradually decreased with advance in age after fifty. Basal serum 17-OHP level was correlated more closely with serum testosterone (T) than cortisol (F), and was suppressed more remarkably after the administration of FM than Dex in both groups. Serum 17-OHP increased with 2 peaks at 6 and 24 hours after HCG injection, and serum 17-OHP response to HCG was significantly greater in the young than in the elderly group. Furthermore, in the aged subjects with low serum T levels, serum 17-OHP response to HCG was remarkably decreased. Within 3 hours after ACTH injection, serum 17-OHP response to ACTH was significantly lower in the elderly than in the young group. On the other hand, 12 hours after ACTH administration, serum 17-OHP response was remarkably greater in the elderly group, and the F/17-OHP ratio was significantly lower than in the young. These data indicate a decline of Leydig cell function in the testis of the elderly male and also suggest the occurrence of enzyme deficiencies, especially impaired C17, 20 lyase activity, in adrenal steroid hormone synthesis with aging. PMID- 6299814 TI - [Prostaglandins may modulate the aldosterone response to ACTH in sodium restricted normotensives]. PMID- 6299815 TI - [A clinical study on the angiotensin I-converting enzyme in human urine. (II) Excretion in hypertensive patients with special reference to the renin angiotensin-aldosterone system]. PMID- 6299816 TI - Glycosidases. (Fucosidases, galactosidases, glucosidases, hexosaminidases and glucuronidase from some molluscs and vertebrates, and neuraminidase from virus). PMID- 6299817 TI - Changes in molecular weight and affinity for adenosine 3',5'-monophosphate of adenosine 3',5'-monophosphate-phosphodiesterase in sea urchin eggs. PMID- 6299818 TI - Some characteristics and hormonal control of thyroidal cAMP-independent protein kinases. AB - Cyclic AMP-independent protein kinases in cytosol from rat thyroid glands were evaluated using histone and casein as exogenous substrates. Compared with other rat tissues, thyroid gland is rich in histone kinases, while its casein kinase activity is lower than that of liver and brain. Thyroidal cAMP-independent protein kinases can be resolved by sucrose gradient ultracentrifugation into two distinct peaks of histone (HKi-1 and HKi-2) and two peaks of casein (CKi-1 and CKi-2) kinases. An intermediate peak of histone kinase activity is only occasionally present. The four main kinase peaks are distinct with respect to several properties: their sedimentation coefficients are significantly different; only one out of the four peaks (CKi-2) can use GTP as substrate; monovalent ions strongly inhibit (50%) light peaks (HKi-1 and CKi-1), while heavy peaks (HKi-2 and CKi-2) are slightly but significantly stimulated (30%); light peaks are very sensitive to thermal inactivation, while heavy peaks are much more resistant. Their reactivity to hormonal action is different: in chronically stimulated glands HKi-2 is selectively and strongly stimulated (240%) while CKi-1 is not changed. In human pathological tissues independent alterations in different kinase entities were observed compared with healthy tissue. In conclusion, the four thyroidal c-AMP-independent protein kinases resolved on sucrose gradient seem to represent distinct entities which are independently and selectively controlled by hormones, and specifically altered in human pathological tissues. PMID- 6299819 TI - Cell type specific inhibition of cAMP phosphodiesterase activity during terminal differential in Dictyostelium discoideum. AB - Cyclic AMP phosphodiesterase (PDE) activity reaches a peak during the aggregation stage of development where it functions to regulate extracellular levels of cAMP. During the subsequent differentiation of the two cell types at the culmination stage, the activity reappears but only in stalk cells. We found that extracts from the culmination stage contained PDE which could be activated by preincubation with Mg2+ and dithiothreitol (DTT), a treatment which is known to release an endogenous inhibitor from the aggregation stage enzyme. When the culmination stage extracts were subjected to chromatography on Biogel P300, two peaks of activity were eluted, PDE-I (Mr greater than 260,000) and PDE-II (Mr 100,000). Treatment of the fractions with Mg-DTT did not affect the low-molecular weight enzyme but caused activation of the high-molecular-weight enzyme and the appearance of a third, intermediate form. Kinetic analysis of the two peaks revealed Km values for cAMP of 2 mM and 10 microM for PDE-I and PDE-II, respectively. We tested the possibility that these forms of the enzyme might be distributed differently in the two cell types by measuring the Km for cAMP and the effect of Mg-DTT treatment on isolated sections of stalk and spore cells. The spore sections contained a high Km form of the enzyme (0.3 mM) which was activated by preincubation with Mg . DTT whereas stalk sections contained a low Km form (3 microM) which was not affected by the activation treatment. We conclude that both cell types contain enzyme protein and that the apparent localization of PDE activity in stalk cells is due to the inhibition of activity in spore cells. PMID- 6299820 TI - Quantitative analysis of cyclic AMP waves mediating aggregation in Dictyostelium discoideum. AB - We have previously reported the detection of cAMP waves within monolayers of aggregating Dictyostelium discoideum cells (K. J. Tomchik and P.N. Devreotes, 1981, Science 212, 443-446). The computer-assisted analysis presented here of the fluorographic images of the cAMP waves reveals (1) all the waves have a consistent width and height; (2) cAMP concentrations within centers of concentric aggregation territories oscillate periodically while at spiral centers the concentration builds up to a plateau value within 2 mm; (3) cells within the region of intersection of two oppositely directed cAMP waves are stimulated to produce more cAMP than those responding to a single wave; (4) cells start to move when the cAMP level begins to increase and cease movement when the peak cAMP concentration reaches the cell. PMID- 6299822 TI - XVIIth International Congress on Herpes virus of Man and Animal: Standardization of Immunological Procedures. Lyon, France, December 7-9, 1981. PMID- 6299821 TI - Differentiation-defective mutants of mouse embryonal carcinoma cells: response to hexamethylenebisacetamide and retinoic acid. AB - We have generated by mutagenesis eight differentiation-defective sublines from three murine embryonal carcinoma (EC) cell lines. These mutants grossly resemble parental cells in the absence of inducers of differentiation. Based upon response to retinoic acid (RA) or hexamethylenebisacetamide (HMBA), the mutants can be grouped into three types: (a) RA-selected cells that lack cellular RA binding protein (cRABP) activity and fail to differentiate in response to RA or HMBA; (b) RA- or HMBA-selected cells that possess cRABP but differentiate poorly, if at all, in the presence of RA or HMBA; and (c) cells originally selected for lack of response to HMBA but which retain cRABP and the ability to differentiate in response to RA. PMID- 6299824 TI - Use of monoclonal antibodies to HSV-1 and HSV-2 for serological analysis of the viral glycoproteins. AB - Herpes simplex virus types 1 and 2 (HSV-1 and HSV-2) specify four major glycoproteins designated gA/gB, gC, gD and gE. Type-specific and cross-reacting monoclonal antibodies to the viral glycoproteins have been used to probe their antigenic determinants. Our studies show the following: (i) Type-specific monoclonal antibodies are suitable reagents for serotyping isolates. Discriminating monoclonal antibodies distinguish antigenic variants among the strains of each serotype. (ii) Glycoproteins gA and gB can not be distinguished from each other with a panel of monoclonal antibodies. gA and gB of HSV-2 made in HEp-2 cells contain type specific domains which are not detected in Vero cells. (iii) Electrophoretic mobilities of glycoproteins gA and gB made in HEp-2 cells are significantly less than those made in Vero cells. Lysates of infected Vero cells contain three lower molecular weight polypeptides that also react with monoclonal antibodies directed to gA and gB. (iv) Immunoadsorbant columns constructed from monoclonal antibodies to glycoproteins gC and gD are useful for purification of the glycoproteins from HSV-1 and HSV-2 infected cell extracts. PMID- 6299823 TI - The differential diagnosis of herpesvirus infections in man and animals. AB - The precise number of human and nonhuman herpesviruses is unknown. It is recognized, however, that the complex antigenic components of this group of viruses and their group interrelationships make specific differentiation of certain herpesviruses extremely difficult with routine serologic procedures. The question of differentiating herpes simplex (H. hominis) infection from B virus (H. simiae) infection frequently requires resolution. When SA8, another primate herpesvirus (principally of African primates) is also involved, differentiation becomes more difficult. The serum neutralization (SN) test, currently the procedure of choice for serologic diagnosis, is not without error. A number of factors control its specificity and variability. Neither EIA, RIA nor FA improves the specificity. Polyarcylamide electrophoresis of the highly lethal B virus indicates the presence of antigens (polypeptides and glycoproteins) distinct from those in other herpesviruses, which may aid in the specific diagnosis, particularly when these antigens are used for the production of monoclonal antibody. DNA fragments have been identified in B virus by restriction endonuclease analysis that appear to be unique for this agent in comparison to human herpesviruses types 1 and 2. PMID- 6299825 TI - [Harmonization of sero-neutralization of Aujeszky's disease virus]. PMID- 6299826 TI - Rapid detection of antibodies to infectious bovine rhinotracheitis by 'macro' and 'micro' ELISA. AB - Two kinds of enzyme-linked immunosorbent assay were evaluated in their ability to detect specific antibodies against Bovine Rhinotracheitis Virus (IBR-IPV). The tests were called MACROELISA and MICROELISA, according to the kind of the solid support used for antigen insolubilization, polystyrene beads and microtitration plates respectively. Partially purified virus was used to coat both beads and plates; a single dilution of examples was tested and protein A linked to peroxidase was employed as enzyme tracer. Quantitative instrumental results from MACROELISA and qualitative visual results from MICROELISA were compared with serum neutralization titers. The results clearly show that ELISA tests are suitable for IBR serologic detection, being sensitive, specific and accurate over serum neutralization method. PMID- 6299827 TI - [ELISA of IBR (infectious bovine rhinotracheitis)--an expression of a 50 percent ELISA titer]. PMID- 6299828 TI - A seroneutralization test in the presence of complement for the serological diagnosis of bovine ulcerative mammillitis. PMID- 6299829 TI - [Detection of intracellular herpes virus antigens by immuno-enzyme procedures]. AB - The antigens of herpes simplex virus types 1 and 2 are detected and localized in vitro in Hep-2 cells by indirect and direct immunoperoxidase. The direct procedure is applied successfully for the diagnosis of genital disease on cytosmears of cervical cells of patients with herpes simplex type 2 infection. With other procedures, as anti-complement immunoperoxidase (ACIP) and anti-IgG immunoperoxidase (AIIP) the Epstein-Barr Nuclear Antigen (EBNA) was detected precisely in EB3, Jijoye and Raji lymphocytes using anti-EBNA antibodies from patients with nasopharyngeal carcinoma or Burkitt lymphoma, in dilution 1:10 1:5120. The indirect immunoperoxidase localized intracytoplasmic early antigen (EA) in 5% of EA--positive Raji cells induced by superinfection of EBV. The viral capsid antigen (VCA) is detected by this method in 5%-10% of virus producing cell lines Jijoye and P3HR-1. The standardized kits of immunoperoxidase procedures for diagnosis of intracellular viral antigens using monoclonal antibodies are under investigation. PMID- 6299830 TI - The use of herpes virus induced dTk as a marker for serological and direct identification of herpes virus infection in man. AB - This report summarizes the essential features of a sensitive deoxythymidine kinase (dTk) assay, which can be used for the diagnosis of Herpes virus infections. A type specific antibody response towards the different Herpes virus dTk isozymes, the Herpes simplex virus (HSV) type 1, type 2, and Varicella Zoster virus (VZV), was found in human sera. The occurrence of dTk antibody in relation to complement fixing (cf) antibody and the state of infection, was found to differ between HSV and VZV. The VZV dTk antibody was only found in patients with Herpes Zoster, while most HSV cf positive individuals had HSV dTk antibody. Absence of dTk in HSV cf positive individuals was related to a "recent primary infection". The type specificity of the dTk isozymes makes these antigens suitable for typing of isolates, and this in combination with the sensitivity of the assay, made it possible to verify and type HSV infection by direct analysis of blister secrete without previous virus isolation. Such results can be obtained within five hours. Combined with conventional isolation procedures the dTk typing assay also detects dTk negative "therapy resistant" strains. The use of the dTk assay for diagnostic purposes is discussed. PMID- 6299831 TI - [Theory and practice of the serology of Epstein-Barr virus]. PMID- 6299832 TI - [Standardization of the indirect hemagglutination reaction in serological diagnosis of cytomegalovirus infection]. PMID- 6299833 TI - Development of sensitive and enzyme-linked immunoassays for herpesvirus antigens and some applications. AB - Sensitive enzyme-linked immunoassays for herpes simplex virus (HSV), cytomegalovirus (CMV) and varicella-zoster virus (VZV) were developed. Both a sandwich technique, using antiviral antibodies from two species to detect the antigen, and an inhibition assay where the sample antigen was incubated with one antiserum, could be used. Around 4-50 ng of viral antigens (measured as protein content) could be detected. The ELISA inhibition technique using type-specific antisera could differentiate between HSV-1 and HSV-2 strains. These could also be distinguished in clinical samples. For CMV, both early and late antigens could be measured as well as the antigenic activity of CMV DNA polymerases. CMV activity in clinical specimens could be detected. The inhibition technique for VZV antigen determination appeared suitable for direct typing of clinical material. No cross reaction with HSV was seen. PMID- 6299834 TI - [Cytomegalovirus (CMV) infection in pregnant women]. PMID- 6299835 TI - The preparation of a reference serum for the determination of IgG and IgM antibodies against cytomegalovirus and its evaluation in a collaborative trial. AB - It is now generally accepted that laboratory data produced by different laboratories using different techniques can be better compared if the results are related to a reference preparation. Even within one laboratory the use of a stable reference preparation helps to reduce the day to day variation of the results. Although this idea has been firmly established for many years the introduction of reference preparations so far has met only with limited success. One of the reasons for this development may be the lack of adequate preparation to be used for this purpose. Recently, diagnostic virology has introduced new serological methods in an effort to make a rapid diagnosis and to differentiate between acute and past infection by the determination of specific antibodies in the IgM, IgA or IgE fraction of the patient's serum. In such a situation, a reference IgM serum of low activity would help to differentiate between persistent or past and recent infection. It was therefore attempted to establish such a reference serum containing antibodies against CMV in the IgM fraction. PMID- 6299837 TI - Marek's disease--survey on vaccination failures. AB - Twelve years after the initial development of efficacious vaccines against Marek's disease--prepared mostly from HVT--and their subsequent use on a large scale, there is a general opinions that failures have been increasing in incidence and in severity. Many factors have been held responsible for these failures, and, though efforts are being made to obviate them, no acceptable solutions have been found sofar in the control of the disease in the field. If mistakes in the preparation, storage handling and administration of vaccine are excepted, because these mistakes, though not infrequent, are easier to prevent, the main causes that must be considered are: 1. The heavy and very early exposure to infection, in houses that are not well disinfected or when birds are reared for many cycles in the same place without cleaning or disinfecting, or in multi age farms, where habits become established, particularly in broilers and pullet rearing, and also for economic reasons. 2. The considerable variability in the susceptibility to MD of different genetic strains of commercial chickens. 3. The appearance of the supposed new biological variants of virus, as recently found in the U.S. The purpose of this study has been the evaluation of the influence of these causes on the more and more frequent outbreaks of MD in our country. While the importance of the genetics and of the degree of and age at exposure has been irrefutably proven, data concerning the appearance of biological variants of MD virus are still short and controversial. PMID- 6299836 TI - Application of molecular genetics to the design of live herpes simplex virus vaccines. AB - In principle, several avenues for attenuation of herpes simplex viruses (HSV) are now available. These include intermixing of HSV-1 and HSV-2 genes by recombination, altering the regulation of gene expression and the deletion of viral genes not required for replication of the virus in permissive cells in culture. Results of analyses of HSV-1 x HSV-2 recombinants and of mutants containing a deletion in a gene expressed early in infection showed a loss of virulence when infected by intracerebral route into adult Balb/c mice. In addition, immunization of the mice by intracerebral route with relatively low doses of virus protected the mice from challenge with high doses (3000 LD50) of virulent virus. The application of genetic engineering to the construction of live vaccines is discussed. PMID- 6299839 TI - Transfection experiments with heated herpes simplex virus and the unreliability of current inactivation controls. AB - Significant infectivity was found in heated and apparently inactivated herpes simplex virus (HSV) types 1 and 2 when cell entry was promoted using the Ca-DMSO transfection technique. Infectivity was quantitated in RSB-1 (rabbit skin fibroblast) and BHK-21 cells, dose-response curves being found to depend upon both cell substrate and thermal side effects. Titres varied with the initial infectivity and cell origin of the virus, and were stable for 6 hours at 56 degrees C, and slightly lower and less stable at 70 degrees C. This report shows that current knowledge concerning the heat-sensitivity of viruses should be revised, and controls for effective inactivation complemented by tests capable of detecting any residual activity of the viral genome. PMID- 6299838 TI - The epidemiology of avian herpesviruses in veterinary medicine. AB - There are ten avian herpesviruses, which have been isolated from eight orders. Six of these are of veterinary importance: Pacheco's parrot disease virus, pigeon herpesvirus, duck plague virus, infectious laryngotracheitis virus, herpesvirus of turkeys and Marek's disease virus. The knowledge on the epidemiology of each virus and the disease it causes is discussed. Features in common to infections with most avian herpesviruses are: infection is persistent in individuals and ubiquitous in populations; virus is shed for long periods of time after infection although in some cases erratically; infection does not necessarily result in disease and at least in some avian herpesvirus infections the incidence of disease is affected by the pathogenicity of the virus; the genetic constitution of the host and stress factors affecting the host. It is concluded that man's interference with the natural history of host species often increases the threat and incidence of disease unless preventive action is taken. PMID- 6299840 TI - Subviral herpes simplex vaccine. AB - An experimental split herpes simplex type 1 (HSV-1) vaccine, virtually free of virus DNA, was prepared from a Nonidet P-40--extract of HSV-1 infected human diploid cells. For determining the contents of neutralizing antigens in these preparations a chromium release inhibition test (CRIT) was developed. The vaccine was gradually shown to be capable of inducing neutralization antibody formation in various animals and conferring protection against intraperitoneal challenge with either type 1 or type 2 viruses in mice. Its administration to mice also markedly restricted the establishment of latent infection following the virus infection. There was an excellent correlation between the immunogenicity of the vaccine and the content of neutralizing antigens as determined by CRIT. PMID- 6299841 TI - Herpes simplex virus subunit vaccine: characterization of the virus strain used and testing of the vaccine. AB - Herpes simplex virus type 1 (HSV-1) used for vaccine production was isolated from a nasal recurrent infection and propagated over a limited number of passages in human diploid cells only. It was designated as HSV-1 BW3 and has been characterized by neutralizing antibodies as a typical HSV type 1 strain. In vitro transformation studies performed with this isolate in mouse or hamster cells revealed only very low, if any, transforming capacity. A preparation of HSV-1 BW3 which can be used as seed lot for vaccine production has been proven to be free of any adventitious agents such as bacteria, fungi mycoplasma or viruses other than HSV-1 BW3. An envelope antigen (EAG) preparation was obtained from purified HSV-1 particles. It was free of detectable viral DNA and could be proven to be efficacious in mouse models against challenge infections with either HSV-1 or HSV 2. The potency of the vaccine was greatly enhanced by the addition of poly inosinic-poly-cytidylic acid complexed with poly L-lysine and carboxymethylcellulose (PICLC) as adjuvant. A single vaccine dose was sufficient to protect mice from morbidity and the fatal outcome of HSV infection, but not from the establishment of latency. Persistent ganglionic infections could be, however, significantly reduced by repeated administration of the vaccine before primary infection. PMID- 6299842 TI - Early experience with "antigenoid" vaccine Ac NFU1(S-) MRC towards prevention or modification of herpes genitalis. AB - The preparation and early clinical experience with "antigenoid" vaccine Ac NFU1(S ) MRC is described. A neutralising antibody response against both type 1 and type 2 herpes simplex virus was stimulated in 19 of 21 and 25 of 60 patients who received the high and low dosage of vaccine, respectively: immunoprecipitating antibody against type-common virus antigen "band II", a virus antigen of major importance in neutralization of herpes simplex virus, was detected in 12 of 19 and 2 of consorts who received the high and low dosage vaccine, respectively. To date, after a mean follow-up period of one year, none of the 42 consorts have contracted herpes genitalis: following a mean follow-up period of 6 months, only 1 patient vaccinated after the initial clinical episode has reported a recurrence of herpetic disease. These results compare favourably with the rates of consort transmission and first year recurrences in unvaccinated subjects. Seventeen of 24 patients with recurrent herpetic disease reported an "improvement" in terms of modification of the frequency, severity or extent of herpetic lesions. There were no important side-effects from vaccination: most patients experienced a degree of minor local reaction at the vaccination site for 24-72 hours and 3 patients reported a mild transient "flu-like" syndrome which disappeared within 48 hours of vaccination. It is intended to subject these data to the scrutiny of a placebo controlled trial with an objective system of clinical assessment. PMID- 6299843 TI - Immune responses to DNA free herpes simplex proteins in man. AB - The data presented confirm in human volunteers our previous observations in animal models. The DNA free HSV 2 subunit vaccine used elicited an antibody and a cell-mediated immune response in 15 subjects without past evidence of HSV 1 or HSV 2 infections and increased the immunity level in 28 subjects suffering from HSV 1 or HSV 2 infections. Although we did not follow a double-blind, placebo controlled protocol our results suggest that the vaccine may reduce the frequency and severity of HSV infections. The time between the recurrences, the pain and the time to complete healing decreased significantly after the vaccination. PMID- 6299844 TI - Immunotherapy with inactivated polyvalent herpes vaccines. PMID- 6299845 TI - Humoral and cellular immune responses of rabbits to sub-viral fractions of human cytomegalovirus. AB - We have raised specific sera against human CMV (strain AD 169) and have tested the immunogenicity of viral envelopes, removed using the non-ionic surfactant Triton X-100. Glycine-buffered extracts of infected MRC-5 cells elicited virus neutralizing antibody to a titre of 1/1600. Incubation of this material also elicited a considerable reaction against host cell antigens. Inoculation of viral envelope material in Freund's complete adjuvant also elicited virus-neutralizing antibodies. Specific in vitro transformation of peripheral blood lymphocytes taken from inoculated rabbits was not always demonstrable, but could be achieved with blood taken from one rabbit up to one year after the initial inoculation of envelope material. PMID- 6299846 TI - Vaccination against human cytomegalovirus with live virus. PMID- 6299847 TI - OKA-SK-RIT Varicella vaccine for "primary" and for booster vaccinations. AB - In seronegative children--healthy and immunocompromised--and in seronegative adults, we have limited but very promising results--no side reactions and good antibody-conversions with clinical protection. Because--in spite of positive humoral antibodies--10 to 20% of people over the age of 50 have a negative cell mediated immune response against VZ and are therefore presumed Zoster patients, VZ-booster vaccinations were done in this age group. A significant increase in humoral antibodies can be induced at least in half of these elderly. In 23 elderly with a negative specific cell-mediated immunity against VZ, a booster vaccination induced a strong positive cell-mediated immune response in 9 and no response in 5. PMID- 6299849 TI - Clinical trial of live attenuated Varicella vaccine in high risk susceptibles--a preliminary report. PMID- 6299848 TI - [Trial of a varicella vaccine in 100 high-risk children]. PMID- 6299850 TI - Marek's disease vaccines. AB - Marek's disease (MD) vaccines have been in almost universal use since the early 1970's and constitute the first effective practical means for the control of any neoplastic disease in man or animals. Five types have been described: 1) attenuated variants of oncogenic strains (Serotype 1) of Marek's disease virus (MDV) produced by serial passage in cultured cells; 2) naturally nononcogenic MDV strains (Serotype 2); 3) nononcogenic herpesvirus from turkeys (HVT, Serotype 3); 4) inactivated cells or cell membrane components from infected cell cultures containing viral-associated antigens; 5) inactivated cells or membrane components from lymphoblastoid MD tumor cell lines containing presumed tumor-associated antigens. HVT vaccines, in either cell-associated or cell-free form, are most frequently used for vaccination of commercial flocks. Both cell-mediated and humoral immune responses are mounted and each may contribute to vaccinal immunity. Probably the most significant responses are against the early (cytolytic) phase of MDV infection, although vaccines apparently also induce a response against tumor-specific antigens as well. PMID- 6299851 TI - [Vaccines against Aujeszy's disease]. PMID- 6299852 TI - [Vaccines and vaccination against infectious bovine rhinotracheitis]. PMID- 6299853 TI - [Trial at vaccination against herpes infection of the pigeon (pigeon herpesvirus 1)]. PMID- 6299854 TI - [Prevention of Marek's disease using 3 vaccines in chickens belonging to different genetic varieties of meat producers]. PMID- 6299855 TI - [Problems in the latency period following vaccination against infectious bovine rhinotracheitis virus (bovid herpesvirus 1)]. PMID- 6299856 TI - Infectious bovine rhinotracheitis: study on the experimentally induced disease and its prevention using an inactivated, adjuvanted vaccine. AB - Experimentally induced IBR was studied in calves. Intranasal challenge enabled reproducible results to be obtained, both from qualitative (clinical aspect) and quantitative points of view (virus excretion, temperature); local and general immunity were also evaluated. This challenge method is useful when studying IBR vaccines. The disease was also experimentally induced by putting healthy animals into contact with diffusor calves. A single injection of inactivated vaccine in oily adjuvant already conferred good protection; it was 100% successful against the experimentally induced disease when administered two times at a 7 or 14 day interval. Immunity obtained was long-lasting and even persisted up to one year. Therefore, this vaccine is advised for vaccination in both contaminated and high risk areas. Results obtained for both safety and potency suggest that this killed vaccine should be used rather than live vaccines. PMID- 6299857 TI - [Feline rhinotracheitis: a purified inactive vaccine and an experimental model]. AB - The medical prophylaxis of feline rhinotracheitis is presently carried out varying success, using three sorts of vaccine: 1. A so-called "attenuated" living virus vaccine, administrable by parental route. 2. A modified living virus vaccine administrable by nasal route. 3. An activated virus vaccine with adjuvant incorporated, administrable by parental route. When this third sort of vaccine was used in contaminated catteries, unfavorable clinical signs were sometimes observed, which resulted in a state of hypersensitivity and even causing excretion of latent virus from convalescent cats which were carriers of Herpes virus. First of all we checked that this was a specific reaction and that it was directly in relation to the viral antigen. Our study was then directed towards the biochemical analysis of the virus and the determination of the sensitizing antigenic fractions and to the immunogenic fraction of the Herpes virus felis. In this paper we report the first results obtained. However, going further than the feline Herpes virus, we think that this is a practical experimental model which could be used in the study of other Herpes virus. PMID- 6299858 TI - Application of a chemically inactivated, adjuvanted vaccine to control abortigenic infection of mares by equine herpesvirus I. AB - A chemically inactivated, adjuvanted vaccine prepared from a virulent strain of Equine herpesvirus I (EHV-I) was used to immunize pregnant Thoroughbred broodmares during a five-year field test designed to determine its safety and efficacy. Each mare in the vaccinated groups received 3 intramuscular injections of vaccine beginning immediately prior to and during the last half of pregnancy. Vaccine was injected at approximately 60-day intervals. The accumulated incidence of EHV-I abortions among vaccinated mares during the field trial period was 1.6/1000 as compared with an incidence of 6.8/1000 in the remainder of the study population. PMID- 6299859 TI - Prophylaxis of CMV infection in bone marrow transplant recipients by hyperimmune CMV gamma-globulin. AB - Prophylactic administration of an intravenous hyperimmune CMV IgG in bone marrow transplant recipients resulted in the complete protection against development of CMV infections and interstitial pneumonia. Incidence of severe graft versus host disease was markedly reduced and there was no mortality in the CMV IgG group. Mortality in those receiving standard supportive therapy was 27%. The source of hyperimmune plasma, method of fractionation and properties of the IgG are described. The effectiveness of this hyperimmune CMV IgG is related to the high content of antiviral antibodies, the dosage, the fact that it is intact, 99% monomeric, and essentially devoid of the IgG3 subclass. The timing of administration together with its effectiveness suggests this preparation might be effective in the treatment of established disseminated CMV infections. PMID- 6299860 TI - Organophosphorous compounds as inhibitors of EBV infection and transformation. AB - Phosphonoformic acid trisodium salt (PF), ethyl diethyl-phosphonoformate (Et-PF), phosphonoacetic acid (PAA), ethyl diethyl phosphonoacetate (Et PAA), ethyl diethyl-2-phosphonopropionate (Et-2-PPA) and ethyl diethyl-3-phosphonopropionate (Et-3-PPA) were tested as inhibitors of EBV virus capsid antigen (VCA) synthesis in B.95-8 cells by immunofluorescence. The most effective inhibitors were: PF, inhibition of 96.74 percent of VCA synthesis at a concentration of 500 microM and PAA, inhibition of 82.05 percent of VCA synthesis at a concentration of 250 microM. Much higher concentrations of the other compounds were needed for inhibition with lower efficiency. Synthesis of EBV nuclear antigen (EBNA) was not inhibited by PF or PAA. However, only PF and PAA inhibited the transformation of human cord blood lymphocytes (CBL) by EBV, as measured by 3H-thymidine uptake, even when added 24 hours after infection. The degree of inhibition of transformation correlated with the concentration. Complete inhibition of CBL transformation by EBV was achieved by 1,000 microM PF and 500 microM PAA, which were noncytotoxic. Removal of PAA from B.95-8 cells cultivated in the presence of high concentrations of PAA for 15 weeks restored VCA synthesis without full concomitant production of transforming EBV. PMID- 6299861 TI - Intertypic superinfection of herpes simplex virus in vivo leads to reisolation of latent viruses with newly acquired lymphotropic properties. AB - The susceptibility of juvenile Tupaias to several strains of temperature sensitive mutants of HSV-1 and 2 was tested. It was found that in all cases the animal survived an infection of 1 X 10(7) PFU of temperature-sensitive mutants which were administered intravenously. In contrast, an infection dose of 1 X 10(3) PFU of wild-type HSV-2 and 1 X 10(2) PFU or HSV-1 was fatal for juvenile Tupaias. Those animals which had initially been infected with the ts-mutants of HSV-1 and/or 2 were protected against a superinfection of a lethal dose of HSV-1 or HSV-2. The state of HSV latency in surviving animals was investigated. The infectious virus was recovered from the spinal cords of those animals which had been initially infected with wild-type HSV-1 or 2 and/or superinfected with HSV-1 or 2. In contrast, the infectious viruses were recovered only from the spleens of those animals which had initially been infected with ts-mutants of HSV-2 and superinfected with HSV-1. These recovered viruses were plaque-purified and analysed in detail. It was found, firstly, that recovered viruses from the spleens of Tupaias lost their pathogenicity and tropism in Tupaias. Secondly, significant changes in the genome of the recovered viruses from the spleen of Tupaias were detected. The results indicate that an intertypic interaction between HSV-1 and 2 occurred in vivo which is responsible for the observed change in organotropism and pathogenicity of the recovered viruses. PMID- 6299862 TI - The role of herpes simplex virus in cervical and vulvar cancer. AB - This report extends the association of herpes simplex virus type 2 (HSV2) with cervical cancer to vulvar carcinoma. In addition to seroepidemiologic observations linking prior HSV2 infection to genital cancer, the new evidence involves the neoplasms themselves. Biopsies of cervical and vulvar cancers were found to contain HSV2-specified antigens. The virus-induced antigens are nonstructural DNA-binding proteins normally present in the nuclei of infected cells; however, in the cancer cells they were found in the cytoplasm. Whole virus structural antigens were not detected in the biopsies. In recent years there has been a parallel rise in prevalence of HSV2 infections and vulvar carcinomas, particularly in women under 40 years of age, reaffirming the association of the virus infection and the neoplasms. PMID- 6299863 TI - A prospective study of the relationship between genital herpes and carcinoma of the uterine cervix. I. Seroepidemiology of herpes simplex virus type 1 and type 2 in women with a previous history of clinically diagnosed genital herpes. AB - The development of a simple, sensitive and reliable enzyme-linked immunosorbent assay (ELISA) for the exact titration of herpes simplex virus type 1 -- and type 2 -- specific antibodies in human serum has made it possible to perform large serological HSV type-specific diagnosis of clinical cases of post and present genital HSV infections. The distribution of HSV type-specific IgG antibody in 473 sera from women with a first episode of symptomatic genital herpes (group A), 602 sera from women with a previous history of symptomatic genital herpes (group B) and 945 sera from non-selected adult Danes (controls, group C) gave the following results: 34 percent of the women in group B had antibodies only to HSV type compared to 6 and 4 percent in group A and C respectively. 64 percent of women in group A were sero-negative, while only 9 and 18 percent sero-negative were found in group B and C respectively. A majority of the sero-negative in group A seroconverted. One third developed antibodies to HSV type 1 and two thirds to HSV type 2. PMID- 6299864 TI - The ontogeny of delta-9-tetrahydrocannabinol responsiveness in the rabbit. AB - An autosomal recessive condition (thc/thc) in our closed colony of New Zealand White rabbits (Uaz:NZW-thc) results in nonfatal, behavioral convulsions following intravenous (i.v.) injections of delta-9-tetrahydrocannabinol (THC) and other psychoactive cannabinoids of marijuana. The ontogeny of the convulsive response was evaluated in potential THC-seizure-susceptible (SS) rabbits from postnatal Days (PN) 15-548. Ages of nonsusceptibility (PN 15-23), partial susceptibility (PN 24-38), and complete susceptibility (PN 39-548) were found. Also, open-field activity was determined in PN 14-25 THC-SS and THC-seizure-resistant (SR) rabbits. Administration of THC during the seizure-insensitive period resulted in genotype-dependent alternations in photocell activity and sprawling. PMID- 6299865 TI - An improved method for the isolation of islets from the beef pancreas. AB - An improved method for the isolation of islets from the beef and pig pancreas is described. The procedure involves the use of strips of Velcro that retain the partially-digested collagen during the isolation of islets by the collagenase technique. The spiny portion of the Velcro is ideally suited to retain the collagen and yet permit the separation of islets from the pancreatic parenchyma. A remarkably high yield of islets has been obtained from the beef and pig pancreas using this procedure. PMID- 6299866 TI - 1,25-dihydroxyvitamin D3 and 24,25-dihydroxyvitamin D3 production by isolated renal slices is modulated by diabetes and insulin in the rat. PMID- 6299867 TI - Effect of poly(ADP-ribose) synthetase inhibitor administration to rats before and after injection of alloxan and streptozotocin on islet proinsulin synthesis. AB - Nicotinamide (10 mmol/kg) and 3-aminobenzamide (1.25 mmol/kg), poly(ADP-ribose) synthetase inhibitors, were injected intravenously to rats either 30 min before the intravenous administration of 12 mg/kg alloxan or 50 mg/kg streptozotocin ("pretreatment") or 5 min after the administration ("posttreatment"). Fifteen minutes after the injection of the diabetogenic agents, pancreatic islets were isolated from the rats and proinsulin synthesis was determined. Proinsulin synthesis was decreased in islets from rats treated with alloxan or streptozotocin. Pretreatment with poly(ADP-ribose) synthetase inhibitors was found to protect against alloxan- or streptozotocin-induced decrease in proinsulin synthesis. By posttreatment with poly(ADP-ribose) synthetase inhibitors, streptozotocin-induced decrease in proinsulin synthesis was also significantly reversed, whereas the decrease induced by alloxan was not. PMID- 6299869 TI - The nosology of genetic peripheral neuropathies in Swedish children. AB - 103 consecutive childhood cases of genetic peripheral neuropathies of heredodegenerative background were collected from Gothenburg from 1973 to 1980. From this series, 63 hereditary motor and sensory neuropathies (HMSN) were distinguished: 31 cases of demyelinating and remyelinating HMSN (HMSN I), 21 (18 families) with an autosomal dominant and 10 with sporadic mode of inheritance and unaffected parents; and 32 cases of neuronal-axonal types (HMSN II), 27 of whom (25 families) had at least one affected, if asymptomatic, parent. In one family, both parents were neurologically and neurophysiologically completely normal. Three cases of uncharacteristic HSN were diagnosed. Among 37 cases with a combined degenerative encephalopathy/myelopathy and a peripheral neuropathy, nine had hereditary spastic paraplegia, six had heredoataxias (three of the Friedreich type), nine had lysosomal storage diseases (five of the Krabbe type), seven had other known inborn metabolic errors and six had biochemically undefined disorders. Progressive neuropathies are important manifestations of a large variety of genetically determined heredodegenerative neurological disorders of infancy and childhood. For classification of HMSN, clinical and neurophysiological examinations are necessary for the index case and for both parents as well. PMID- 6299868 TI - Peripheral neuropathy in rats induced by insulin treatment. AB - The effect of sustained insulin-induced hypoglycemia on peripheral nerve function and structure was examined in rats. After a period of hypoglycemia (less than 2.5 mmol/L) of at least 72 h, axonal degeneration and reduction of the maximal amplitude of the evoked muscle action potential occurred, the two abnormalities being correlated negatively (r = -0.99, 2P = 0.00097). One of five rats developed paresis of both hindlegs as well as nerve damage and perikaryal alterations of lower motor neurons. PMID- 6299870 TI - Research on AIDS. PMID- 6299871 TI - Comparative carcinogenicities of model compounds of metabolically activated N,N dibutylnitrosamine in rats. PMID- 6299872 TI - Effects of adriamycin and macrophage-toxic agents on lymphoma growth in lethally irradiated mice. PMID- 6299873 TI - Cholangiocarcinoma associated with a type 2 choledochal cyst. PMID- 6299874 TI - Effect of colonic perfusion with sulfated and nonsulfated bile acids on mucosal structure and function in the rat. AB - A single-pass perfusion system was used in conscious restrained rats to measure changes in water and electrolyte transport and in protein and deoxyribonucleic acid output after perfusing 5-15-mM solutions of sulfated or nonsulfated bile acids through the colon. Perfusion with 5 mM nonsulfated deoxycholic acid or chenodeoxycholic acid changed net water and sodium absorption to net secretion, provoked marked increases in protein and DNA output into the perfusion effluent, and caused microscopic mucosal damage. In contrast, perfusion with 5 and 15 mM sulfated deoxycholic acid or with 5 mM sulfated chenodeoxycholic acid had no effect on water and electrolyte transport and caused only modest changes in protein and DNA output. To see whether or not sulfated bile acid could prevent the effect of its nonsulfated parent compound on colonic structure and function, perfusion with a mixture of 5 mM nonsulfated and 10 mM sulfated deoxycholic acid was performed. This produced net secretion of water and sodium together with less marked increases of protein and deoxyribonucleic acid output and less pronounced microscopic mucosal damage than was seen after 5 mM nonsulfated deoxycholic acid alone. Finally, 5 mM nonsulfated cholic acid had no effect on water or sodium transport, but 5 mM sulfated cholic acid, with one alpha-hydroxyl group masked and two alpha-hydroxyl groups "exposed," reduced water transport. These results suggest that sulfation prevents the cathartic effect of alpha-dihydroxyl bile acids in the colon. PMID- 6299875 TI - Portal hypertension and gastric mucosal injury in rats. Effects of alcohol. AB - The present study was performed primarily in order to determine whether gastric mucosa of rats with portal hypertension has different functional and histologic features when compared with controls, and second to quantitate and compare morphologic and functional changes after exposure to topical ethanol. Portal hypertension was produced by staged portal venous occlusion, and in these animals portal pressure was 32 +/- 2 cm saline compared with 18 +/- 2 cm in sham-operated controls (p less than 0.005). Before ethanol, portal hypertensive rats compared with controls had significantly higher luminal pH (2.9 +/- 0.3 vs. 1.9 +/- 0.1), increased H+ back-diffusion (loss of 138 +/- 10 vs. 57 +/- 16 microEq H+/h), lower potential difference (8 +/- 1 mV lower than controls), and extensive submucosal edema (submucosal thickness 325 +/- 25 vs. 138 +/- 18 micrometers). After 3 h of exposure to 2 ml intragastric absolute ethanol, the area of macroscopic hemorrhagic mucosal injury was significantly greater in portal hypertensive rats than in controls (34.0 +/- 8.7% vs. 7.6 +/- 2.1%), confirmed histologically by the greater number of deep hemorrhagic necrotic lesions and extent of mucosal length involved. Furthermore, after ethanol, portal hypertensive rats compared with controls had significantly increased gastric volume (14.4 +/- 1.5 vs. 8.3 +/- 0.6 ml), Na+ (86.6 +/- 8.0 vs. 64.6 +/- 8.0 mEq/L), pH (7.1 +/- 0.3 vs. 4.3 +/- 0.4), H+ back-diffusion (loss of 309 +/- 41 vs. 207 +/- 33 microEq H+/h), and protein and blood loss (100% increases over controls). These results indicate that gastric mucosa of portal hypertensive rats has distinctive functional and histologic abnormalities that can explain its increased susceptibility to erosive injury after ethanol. This study quantitatively confirms in an animal model the clinical observations that portal hypertension may predispose to severe gastric mucosal injury. PMID- 6299876 TI - Hepatic adenoma and focal nodular hyperplasia: clinical, pathologic, and radiologic features. AB - Based on strict predefined histologic criteria, we identified 23 patients with hepatic adenoma and 41 patients with focal nodular hyperplasia seen at this institution between 1961 and 1980. Patients with hepatic adenoma were young and 91% were female. When a reliable history was available, 89% had used oral contraceptives; 53% presented acutely or with pain. Eleven of 11 radionuclide scans were abnormal; 15 of 15 angiograms showed a hypervascular mass with 7 of 15 showing areas of hypovascularity. Eighteen resections were performed with one operative death. Two tumors contained areas of unequivocal hepatocellular carcinoma. Fifteen of 18 patients followed for 82 +/- 11 mo were living and well and had discontinued oral contraceptives. Focal nodular hyperplasia patients were older, 88% were female, and 58% had used oral contraceptives. Their lesions were discovered accidentally. Seven of 12 radionuclide scans demonstrated voids, while 13 of 13 angiograms showed hypervascular lesions with no areas of hypovascularity. Seventeen tumors were resected. Twenty-three of 24 patients followed for 45 +/- 7 mo were living and well. One died of nonhepatic causes. Based on the findings of this review, we believe that if the clinical suspicion of hepatic adenoma or focal nodular hyperplasia is strong, elective laparotomy for diagnosis is usually the best approach. Hepatic adenoma should be resected if technically feasible. Intraoperative wedge biopsy is appropriate for focal nodular hyperplasia. PMID- 6299877 TI - [How 67 carcinomas of the breast with a maximal microscopic diameter of 10 mm were recognized]. AB - 67 breast cancers with a maximum diameter of up to 10 mm on the histological section were retrospectively analysed according to the primary situation for medical consultation and the way to cancer diagnosis. 46 women presented themselves with clinical signs. 19 women had an anamnestical risk, whereas only two women were without any risks or symptoms. 34 tumourspecific clinical signs are followed by 25 tumourspecific mammograms. Eight small invasive carcinomas were detected only under the microscope, two of them in mastectomy specimens. Even small cancers, therefore, are mostly diagnosed from the patients themselves. The contribution of mammography is tremendous. Intensified systematic histological examination of biopsies and mastectomy specimens is mandatory especially in high risk patients. PMID- 6299879 TI - [Cloning the operon genes of riboflavin biosynthesis in Bacillus subtilis on plasmid vector pBR322 in Escherichia coli]. AB - The operon for riboflavine biosynthesis of Bacillus subtilis wild type and its operator-constitutive derivative have been cloned in Escherichia coli cells on the plasmid pBR322 vector. The plasmids constructed were able to transform strains of E. coli and Bac. subtilis from Rib- to Rib+ phenotype. A DNA insert into the EcoRI site of pBR322 causes a decrease in tetracycline gene expression. The operator of the riboflavine operon of Bac. subtilis does not participate in regulation of the operon expression in E. coli cells. PMID- 6299878 TI - Human mitochondrial DNA variation and evolution: analysis of nucleotide sequences from seven individuals. AB - We have analyzed nucleotide sequence variation in an approximately 900-base pair region of the human mitochondrial DNA molecule encompassing the heavy strand origin of replication and the D-loop. Our analysis has focused on nucleotide sequences available from seven humans. Average nucleotide diversity among the sequences is 1.7%, several-fold higher than estimates from restriction endonuclease site variation in mtDNA from these individuals and previously reported for other humans. This disparity is consistent with the rapidly evolving nature of this noncoding region. However, several instances of convergent or parallel gain and loss of restriction sites due to multiple substitutions were observed. In addition, other results suggest that restriction site (as well as pairwise sequence) comparisons may underestimate the total number of substitutions that have occurred since the divergence of two mtDNA sequences from a common ancestral sequence, even at low levels of divergence. This emphasizes the importance of recognizing the large standard errors associated with estimates of sequence variability, particularly when constructing phylogenies among closely related sequences. Analysis of the observed number and direction of substitutions revealed several significant biases, most notably a strand dependence of substitution type and a 32-fold bias favoring transitions over transversions. The results also revealed a significantly nonrandom distribution of nucleotide substitutions and sequence length variation. Significantly more multiple substitutions were observed than expected for these closely related sequences under the assumption of uniform rates of substitution. The bias for transitions has resulted in predominantly convergent or parallel changes among the observed multiple substitutions. There is no convincing evidence that recombination has contributed to the mtDNA sequence diversity we have observed. PMID- 6299880 TI - [Restriction-modification systems in Bacillus strains related to Bacillus subtilis]. AB - 127 strains of bacilli sensitive to different phages of Bacillus subtilis were isolated from the soil of Moscow and its country-side. In 6 strains, restriction and modification systems were discovered which differed from these previously described for Bac. subtilis BsuR system. Two strains has identical restriction modification systems, and one strain possessed two different systems. Using DNA from all 6 strains, it was possible to transform competent cells of Bac. subtilis RUB834. Two of these 6 strains could serve as recipients in transformation and transfection experiments. PMID- 6299881 TI - [Genetics of nephroblastoma. II. The incidence of developmental defects and dysplastic and asymmetrical traits in children with nephroblastoma]. AB - The frequency of developmental defects and displasticity characters in children with nephroblastoma was determined in two groups--with early, up to 2 years manifestation of the tumor (ENB), in 40 patients, and with advanced nephroblastoma (ANB), in 59 patients (manifestation of the disease after 2 years). The data were correlated with control groups adequate for age (130 children). The frequency of serious developmental defects was higher in ANB group (20%) than in ENB group (7%). Over a half of developmental defects were hemihypertrophy and doubling of organs. One case of a child with the combination of nephroblastoma and Smith-Lemli-Opitz syndrome was defined. In ANB group an increased frequency of asymmetry in the conjugate organs (foot, hand, middle finger, ears) was found. Its direction is correlated with tumor localization (tumor site). In ENB group no analogous effect was shown. The data obtained present phenotype characteristics of groups with early and late manifestation of the disease which are probably different in their genesis and thus, their identification is necessary for the adequate medicogenetic consultation (examination). PMID- 6299882 TI - Characterization of the cloned terminators tR1, tL3 and tI and the nut R antitermination site of coliphage lambda. PMID- 6299883 TI - A physical map of the ribosomal DNA repeat unit of Aspergillus nidulans. AB - The ribosomal DNA repeat unit of Aspergillus nidulans has been cloned in pBR322 and a restriction map constructed. The genes coding for the 17S, 5.8S and 25S rRNAs are found in blocks separated by a 1.7 kb spacer region, with the 5.8S RNA gene lying between the genes for the two larger RNAs. The total length of the repeat unit is 7.7 kb. The 5S rRNA is not present in the repeat unit. PMID- 6299884 TI - Cyclization of linear chimeric plasmids in vivo by a novel end-to-end joining reaction or by intramolecular recombination: one of the products contains a 147 bp perfect palindrome stable in Escherichia coli. PMID- 6299885 TI - Nucleotide sequence and evolution of the rat mitochondrial cytochrome b gene containing the ochre termination codon. AB - The nucleotide sequences of the genes for cytochrome b and three potential transfer RNAs (tRNAPro, tRNAThr and tRNAGlu) in cloned rat mitochondrial DNA were determined. The derived amino acid sequence of the cytochrome b protein from the light strand indicated that the C-terminal amino acid is asparagine and the ochre termination codon is encoded in the DNA, in contrast to the the lack of termination codon in the reading frame of human [Anderson et al., Nature 290 (1981) 457] or mouse [Bibb et al., Cell 26 (1981) 167] mitochondrial DNA. The first ATG codon of the cytochrome b gene was spaced five nucleotides from the 5' end of the tRNAGlu gene on the heavy strand. There was a single nucleotide spacing between the termination codon of the cytochrome b gene and the 5' end of the tRNAThr gene in the light strand. There was also a single nucleotide spacing between the 3'-end of the tRNAThr gene and the 3'-end of the tRNAPro gene on the heavy strand. The amino acid and nucleotide sequences of the cytochrome b genes of mammals and yeast [Nobrega and Tzagoloff, J. Biol. Chem. 255 (1980) 9828] were compared to reveal structural differences in two very different species. At the same time, amino acid substitutions in particular regions of the mammalian gene corresponding to the exon-intron boundaries in the yeast gene were noted. These genetic features are discussed in relation to the extreme compression of genetic information in the mammalian mitochondrial genome as related to the evolution of the gene organization and its sequence. PMID- 6299886 TI - Fusion of restriction termini using synthetic adaptor oligonucleotides. AB - A method is proposed for linking a DNA fragment possessing a 5' single-stranded extension to one carrying a 3' extension. Synthetic oligonucleotide adaptors can be used to (i) change the site specificity at the termini of a fragment generated by restriction enzyme cleavage and (ii) simultaneously dephosphorylate the extremities of a DNA molecule to prevent recircularisation and allow positive selection for recombinant DNA molecules. PMID- 6299887 TI - Cloning of the modification methylase gene of Bacillus centrosporus in Escherichia coli. AB - The gene specifying a sequence-specific modification methylase of Bacillus centrosporus has been cloned in Escherichia coli using the restriction endonuclease HindIII and the plasmid pBR322. The selection was based on detection of new methylation properties rendering recombinant plasmids carrying the methylase gene nonsusceptible to BcnI endonuclease cleavage. The presence of a 3.2-kb HindIII fragment in either orientation conferred BcnI resistance on the recombinant plasmids. These results suggest that the BcnI methylase gene is expressed in E. coli under the control of a promoter located on the cloned fragment. The relative level of BcnI methylase enzyme in E. coli was similar to that in B. centrosporus. The recombinant clones do not exhibit any BcnI restriction-endonuclease activity. PMID- 6299888 TI - Restriction mapping and molecular cloning of adenovirus type 4 (subgroup E) DNA. AB - The locations of thirty restriction endonuclease cleavage sites were determined on the genome of adenovirus type 4 (Ad4), the sole member of the subgroup E adenovirions. The restriction endonucleases Bg/II, EcoRI, HindIII, HpaI, KpnI, SalI, and XbaI cut Ad4 DNA 10, 3, 2, 3, 5, 5 and 3 times, respectively. Orientation of the linear Ad4 map with respect to left and right molecular ends was accomplished by taking advantage of the limited sequence homology between Ad2 and Ad4. Ten non-overlapping fragments of Ad4 DNA representing 98% of the genome, map units 1.6 to 99.6, have been cloned into the plasmid vector pKC7. PMID- 6299889 TI - Expression of the Eco RI restriction-modification system and the construction of positive-selection cloning vectors. AB - The genes encoding the Eco RI restriction-modification (R/M) system have been separately cloned onto compatible plasmids. We have shown that the Eco RI restriction gene is expressed in the total absence of methylase enzyme and confirmed that a temperature-sensitive mutant is defective in Eco RI modification activity at higher temperatures. Insertion of transcriptional terminators into the restriction gene had no detectable effect on Eco RI modification activity. This strongly suggests that a separate promoter exists for the methylase gene. Analysis of the published sequence shows that the methylase gene promoter may overlap with the COOH-terminal region of the endonuclease structural gene. The temperature-sensitive Eco RI system has been exploited in the construction of two plasmid cloning vectors, pLV57 and pLV59, which can be used to select positively for transformants bearing recombinant plasmids; cloning of a DNA fragment into pLV57 or pLV59 at the unique HindIII, Bg/II, or PstI sites inactivates the Eco RI restriction gene and permits the hybrid plasmid to survive at 37 degrees C. The temperature-sensitive modification activity of these vectors should also facilitate the introduction of Eco RI linkers into DNA cloned in this way. PMID- 6299890 TI - Construction and analysis of in vivo activity of E. coli promoter hybrids and promoter mutants that alter the -35 to -10 spacing. PMID- 6299891 TI - A simple method for shortening a plasmid genome using a system of plasmid cointegration mediated by a Tn3 mutant. PMID- 6299892 TI - Preparation of product-specific antisera by gene fusion: antibodies specific for the product of the yeast cell-division-cycle gene CDC28. AB - Antisera with specificity for the product of a yeast cell-division-cycle (CDC) gene were prepared by immunizing rabbits to a novel hybrid polypeptide. A segment of the yeast gene CDC28 was fused to the Escherichia coli lacZ gene, which encodes beta-galactosidase, by insertion of yeast sequences into the plasmid pBGF1. pBGF1 contains the lac promoter-operator and most of the lacZ gene. An EcoRI site, 16 codons upstream from the carboxyterminus of the beta-galactosidase coding region, served as a convenient splicing site for the heterologous sequences. To insure that an open reading frame be maintained between the two gene segments for some portion of the fusions, the CDC28-encoding segments were first subjected to limited digestion with nuclease BAL31 to produce random junction points. A hybrid polypeptide encoded by such a continuous open reading frame was purified from E. coli by preparative SDS-polyacrylamide gel electrophoresis and used to immunize rabbits. The resulting antisera were shown to have specificity for CDC28 gene product synthesized by cell-free translation of yeast mRNA. PMID- 6299893 TI - Identification of sequences necessary for packaging DNA into lambda phage heads. AB - Several species of DNA molecules are packaged into lambda phage heads if they carry the region around the cohesive end site of lambda phage (cos lambda). The minimal functional sequence around cos lambda needed for packaging was examined by cloning in pBR322. The results showed that the minimal region contained 85 bp around cos lambda; 45 bp of the left arm of lambda phage and 40 bp of the right arm. A 75-bp region located to the right of the minimal region seems to enhance packaging. A 223-bp fragment containing these regions can be used as a portable element for plasmid DNA packaging into lambda phage heads. Plasmid ppBest 322, a derivative of pBR322 carrying this portable packager and both amp and tet genes, was constructed. This plasmid is useful for cloning of large DNA fragments. PMID- 6299894 TI - Construction and characterization of E. coli promoter-probe plasmid vectors. III. pBR322 derivatives with deletions in the tetracycline resistance promoter region. AB - Deletions of the promoter region for the tetracycline-resistance (Tcr) gene(s) of pBR322 were constructed in order to generate new promoter-probe plasmid cloning vectors. The deletions were constructed in vitro by exonuclease digestion at the HindIII site and blunt-end ligation of the digestion products. Plasmids which lost the HindIII site but retained the EcoRI site carried deletions ranging from 5 to 60 bp. Some of the plasmids lacked the nucleotide sequences required for initiation of transcription from the Tcr promoter and "anti-Tcr" promoter. Three of the promoter-deletion plasmids (containing deletions of 5-29 bp) formed tight binding complexes with RNA polymerase in vitro, despite their tetracycline sensitive phenotype. One deletion plasmid, pPV33, retained three out-of-phase stop codons located between the promoter-cloning site (EcoRI) and the translational start codon for the tetracycline resistance gene. These features give pPV33 several advantages over previously described promoter-cloning vehicles. PMID- 6299896 TI - A bacteriophage lambda vector for cloning with BamHI and Sau3A. AB - A phage lambda cloning vector has been constructed which contains a single site for the restriction endonuclease BamHI. Since Sau3A and Bg/II produce the same cohesive ends as BamHI, this vector can also be used to clone DNA fragments generated with either of these enzymes. We have used this vector to construct an Escherichia coli library using partial digestion with Sau3A. This vector will be most useful for applications requiring genetic analysis of cloned E. coli genes. PMID- 6299895 TI - Construction and characterization of the chloramphenicol-resistance gene cartridge: a new approach to the transcriptional mapping of extrachromosomal elements. PMID- 6299897 TI - Isolation and characterization of nuclear genes coding for subunits of the yeast ubiquinol-cytochrome c reductase complex. AB - Nuclear genes coding for the Mr 17 000, 14 000 and 11 000 subunits of the ubiquinol-cytochrome c reductase complex (complex III) in yeast have been isolated from a clone bank of yeast nuclear DNA by use of a mRNA hybridization competition assay. This is based on our observations that levels of mRNAs for these subunits are much reduced during glucose repression and in cytoplasmic petite mutants and the procedure should be of general application for the isolation of other inducible or repressible genes coding for mRNAs present at low levels in the cell. A first characterization of the clones is presented. The genes are not closely linked in the genome and those coding for Mr 14 000 and 11 000 subunits are present in unique genomic environments, which suggests that there are only single copies of each in the nuclear genome. PMID- 6299898 TI - Cloning of the am (glutamate dehydrogenase) gene of Neurospora crassa through the use of a synthetic DNA probe. AB - In a previous study the alteration in the amino acid sequence of Neurospora crassa NADP-specific glutamate dehydrogenase (GDH) resulting from two mutually compensating frameshift mutations was used to deduce the first 17 nucleotides of the coding sequence of the am gene. In the work reported here, a synthetic 17-mer corresponding to the deduced sequence was shown to hybridize strongly to a 9-kb HindIII fragment from N. crassa wild-type DNA but not to any corresponding fragment from the DNA of a mutant strain known to be deleted for most or all of the gene. Wild-type HindIII fragments were fractionated for size and a fraction centering around 9 kb was cloned in vector lambda L47. Two clones carrying the strongly hybridizing fragment were identified. The hybridization to the 17-mer was localized within a 2.7-kb BamHI fragment and, within this, to a 700-bp BamHI Bg/II subfragment. 5' end-labelled polyadenylated RNA isolated from wild-type mycelium hybridized to the 2.7-kb BamHI fragment and not appreciably to flanking fragments. The partial sequence analysis of the BamHI-Bg/II fragment has confirmed that the 17-mer probe matches the coding sequence at the 5' end of the gene and has also revealed an intervening sequence 67 bp in length, interrupting codon 15. Both the 9-kb HindIII fragment and the 2.7-kb BamHI fragment have been shown to be capable of transforming the deletion mutant to prototrophy and ability to produce GDH. Analysis of one transformant showed that the am gene was integrated, together with a part of the long arm of the lambda vector, at an unusual locus. This transformant, in which the am gene does not show its normal linkage to the linkage group 5 marker inl, was found to produce GDH to about 20% of the normal level. PMID- 6299899 TI - KpnI families of long, interspersed repetitive DNAs associated with the human beta-globin gene cluster. AB - KpnI families of long, interspersed repetitive DNAs are ubiquitous repetitive elements that occur in tens of thousands of copies in primate genomes. KpnI 1.2, 1.5 and two different KpnI 1.8-kb families were found within and flanking a 6.4 kb repeat beginning at 3 kb, 3' from the human beta-globin gene. Thus, six different types of KpnI families have now been identified, and four of these are found next to each other in a specific 6.4-kb repeat. Clones of the distinct KpnI families were hybridized to clones of the 6.4-kb repeat and adjacent sequences encompassed within some 17.6 kb of DNA lying 3' to the beta-globin gene cluster. The four KpnI families appear to make up the entire length of the 6.4-kb repeat. The linear order of the various cloned KpnI sequences in the repeat is 5' pBK(1.8)26-pBK(1.5)54-pBK(1.2)11-pBK(1.8)11-3'. KpnI 1.2-kb sequences were also detected downstream from the 6.4-kb repeat. As in the case of the KpnI 1.2 and 1.5-kb families, the two KpnI 1.8-kb sequence families described here each hybridized with about 15% of all plaques in two independently generated human genome libraries. PMID- 6299900 TI - New orientations of ancestral, "long interspersed repeated sequences" (LINES) in human DNA. AB - Three sets of long, interspersed repeated sequences (LINES) are described in human DNA. Each set contains two cleavage sites for the restriction endonuclease, XbaI. One set, called the Xba 850 LINES was detected only in gibbons, apes and man but is related in sequence to a more ancestral LINES family, the Kpn 1200 LINES, and in fact some Xba 850 LINES members retained the ancestral spacing of KpnI cleavage sites. The facts that the Xba 850 LINES appear as a subset of the Kpn 1200 LINES and vice versa and that the Xba 850 LINES are restricted to a smaller phylogenetic group than the Kpn 1200 LINES prompted the speculation that the Xba 850 LINES originated by a relatively recent amplification of one or a few Kpn 1200 LINES sequences. PMID- 6299901 TI - Genetic transformation of Saccharomyces cerevisiae with single-stranded circular DNA vectors. PMID- 6299902 TI - Development of cloning vehicles from the Streptomyces plasmid pFJ103. AB - A 20-kb plasmid, pFJ103, was isolated from a strain of Streptomyces granuloruber. A restriction endonuclease map of the plasmid was constructed. A Streptomyces gene that specifies resistance to the antibiotic thiostrepton was subcloned into Escherichia coli plasmid pBR322, inserted into pFJ103 and transformed into Streptomyces ambofaciens protoplasts. Two classes of transformants were obtained. One carries the pFJ104 plasmid consisting of the entire pFJ103 with the 1.8-kb thiostrepton resistance gene insert. The other carries the pFJ105 plasmid consisting of the 2.9-kb replicon segment of pFJ103 with the same thiostrepton resistance insert. A gene for neomycin resistance together with the entire E. coli pBR322 plasmid were cloned into pFJ105. The resulting E. coli-Streptomyces bifunctional vector, pFJ123, transformed both E. coli and Streptomyces. The small size of pFJ105, its ease of isolation, and efficient transformation of Streptomyces protoplasts establishes it, and its derivatives, as useful plasmid cloning vehicles for fundamental and applied studies. PMID- 6299903 TI - Direct selection of recombinant plasmids in Bacillus subtilis. AB - A system is described which permits the direct, positive selection of recombinant plasmids in Bacillus subtilis. This system relies on the plasmid pBD214 which confers chloramphenicol (Cm) resistance and carries a thy gene, and on BD393, a highly competent B. subtilis thyA thyB host. Thy- strains are resistant to trimethoprim (Tmp), and Thy+ strains are sensitive. Inactivation of the pBD214 thy determinant by insertion of a DNA fragment permits selection of Cmr Tmpr clones, all of which carry recombinant plasmids. This insertional inactivation can be accomplished using the unique EcoRI, BclI, PvuII, or EcoRV sites, all of which are located within the thy gene on pBD214. Some properties of this selective system are described, and its uses for molecular cloning are discussed. PMID- 6299904 TI - Restriction map of the hepatitis B virus DNA cloned in Escherichia coli. AB - A plasmid carrying the complete genome of hepatitis B virus (HBV) inserted into the BamHI site of the pBR322 plasmid vector has been constructed. The physical map of HBV DNA established for 13 restriction endonucleases allows to conclude that the cloned DNA is similar, but not identical to the HBV DNA of ayw subtype. PMID- 6299906 TI - An assessment of sequential measurements of immune complex levels in ovarian cancer patients with respect to clinical progress. AB - An evaluation of immune-complex levels as an additional parameter in the clinical follow-up of ovarian cancer patients is described. These patients were treated aggressively with monthly chemotherapy following surgery. Sequential measurements of immune-complex levels have been carried out concurrently and clinical findings were recorded in a uniform manner suitable for comparison with the levels of immune complexes. The relationships between changes in tumor mass and changes in levels of immune complexes were investigated. A significant relationship between decreasing immune-complex levels and simultaneously decreasing tumor mass was found. Chemotherapy was associated with a decrease in immune-complex levels which increased again within 1 month. These data did not support an effective role for immune-complex levels in the evaluation of these patients. PMID- 6299905 TI - Role of bone scanning for evaluation of carcinoma of the cervix. AB - The value of X-ray studies and bone scan studies of cervix cancer patients was evaluated by a review of patient charts and bone scans or X-ray studies. No in situ cancer patients had positive studies. A low frequency of positive X-ray studies was observed ranging from 1% with stage I to 10% with stage IV disease. Recurrent carcinoma had 10% positive studies. Bone scans correlated well with positive X-ray studies but 10% false-positive scans were seen, mostly osteoarthritis. The bone scan was much more sensitive and appeared to detect an earlier stage lesion, usually associated with symptoms which with radiation and other treatments could follow a more indolent course than the patient with a positive X-ray study. The present study evaluates the appearance and value of these studies, the frequency and circumstances under which useful information was obtained, and what type of survival followed the establishment of the diagnosis. PMID- 6299907 TI - Genital warts and cervical cancer. II. Is human papillomavirus infection the trigger to cervical carcinogenesis? AB - Evidence of a biologically significant association between subclinical papillomavirus infection (SPI) and cervical neoplasia raises the question of whether this is a causal relationship. Human papillomavirus (HPV) infection of metaplastic epithelium in the cervical transformation zone is relatively common, producing latent infection in susceptible persons. The epidemiological characteristics of SPI and cervical cancer are essentially identical and there is a strong clinico-pathological association between condylomas and anogenital carcinoma. Tissue culture cells have been transformed from a normal to a neoplastic phenotype by animal papillomaviruses, and there is preliminary data reporting upon the successful identification of HPV genomic sequences in tumor cells. SPI commonly coexists with foci of cervical intraepithelial neoplasia (CIN). Areas of apparent transition are seen, and these two lesions are linked by a discernible spectrum of morphologic change. Such circumstantial evidence gives biological plausibility to the suggestion that HPV may be a cervical carcinogen. It is postulated that cervical neoplasia arises by progression from benign viral hyperplasia, through varying stages of koilocytotic atypia with associated dysplasia, to unremarkable carcinoma in situ. Invasion is presumed to reflect the emergence of an aggressive heteroploid clone, an age-related decline in host immune surveillance or an interaction of both factors. PMID- 6299908 TI - [Clinical use of antiandrogens in the female]. PMID- 6299909 TI - [Principles and limits of breast-saving therapy of small breast cancers]. PMID- 6299910 TI - [Tumor markers and their significance in gynecologic oncology]. PMID- 6299911 TI - Effects of dietary broccoli and butylated hydroxyanisole on liver-mediated metabolism of benzo[a]pyrene. AB - Groups of male Wistar rats were given either a basal diet or diets supplemented with 10 or 25% broccoli or 0.8% BHA. Liver fractions were assayed for cytochrome P-450, for aryl hydrocarbon hydroxylase (AHH), glutathione-S-transferase and epoxide hydrolase activities and for benzo[a]pyrene (BaP) metabolism. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis of liver microsomes was also performed. Mean relative liver weight in the BHA group was significantly higher than that of the control and 10% broccoli groups but not significantly higher than that of the 25% broccoli group. Gel electrophoresis of liver microsomes indicated a diet-dependent variation in intensity in a band that corresponded in mol wt to those of certain cytochrome P-450s. Diet-dependent increases (20-90% over control) in cytochrome P-450 and in the activities of AHH, glutathione-S transferase and epoxide hydrolase were observed in livers from rats given broccoli-supplemented diets. Except for AHH activity, such increases also occurred in the group fed BHA. Analysis of BaP metabolites revealed that the proportion of 4,5-diol formed relative to the major diols identified was unchanged in the broccoli- or BHA-treated groups relative to the control group. The proportion of 9,10-diol formed was unchanged in the broccoli-fed groups but was significantly higher in the BHA group than in the control group. The proportion of cis and trans-7,8-diol formed was unchanged in both broccoli-fed groups but was significantly lower in the BHA group. In comparison with the control group, the ratio of phenol I (comprising primarily 9-OH-BaP) to total phenols (primarily 9-OH and 3-OH) was significantly decreased by about 30% in the 25%-broccoli group and by about 70% in the BHA group. Qualitative differences in the phenol-II peak (comprising 3-OH and 7-OH phenols) were also observed between samples from the controls and those of 25%-broccoli- and BHA-fed rats. The implications of these findings are discussed with respect to the effects of broccoli and BHA on benzo[a]pyrene toxicity. PMID- 6299912 TI - Zeolithe A--A phosphate substitute for detergents: toxicological investigation. AB - Tests on Zeolithe A, a sodium aluminium silicate developed as a substitute for phosphates in detergents, were designed to investigate the safety of exposure to the material, or to detergents containing it, either under industrial conditions encountered during manufacturing processes or as a consequence of domestic use. The test programme included oral studies (acute, subchronic and long-term carcinogenicity tests and absorption measurements), and dermal, ocular and inhalation studies on the silicate alone and on appropriate detergent formulations, as well as studies of possible silicogenic activity and metal complexing potential and measurements of dust generation and particle-size distribution. These studies did not produce any evidence to suggest that levels of domestic and industrial exposure resulting from the projected use of Zeolithe A in detergents would present any hazard to health. Zeolithe A did not induce silicotic tissue reactions and when incorporated into detergent formulations did not increase the liberation of fine dusts. PMID- 6299913 TI - [Involvement of the colon in gastric and pancreatic processes]. AB - Using representative case studies the diagnostic problems of colonic infiltration in cases of gastric and pancreatic neoplastic or inflammatory lesions are discussed. Typical signs in both barium enema and ultrasonography lead to the detection of secondary bowel wall lesions. Characteristic intraperitoneal pathways of spread and roentgen-morphologic features are important in the attempt to differentiate inflammatory from malignant infiltrations. In these cases ultrasonography, showing a diffuse thickening of gastric or bowel wall, is a useful additional diagnostic tool, which gives further information about topographical relations in the abdomen. PMID- 6299914 TI - [Emphysematous cholecystitis caused by Clostridium perfringens. Sonographic and computer tomographic findings]. PMID- 6299915 TI - [Congenital cytomegalic inclusion disease in cranial computerized tomography in an infant]. PMID- 6299916 TI - HBeAg/anti-HBe system and development of primary hepatocellular carcinoma in patients with HBsAg-positive liver cirrhosis. AB - The significance of the HBeAg/anti-HBe system and other HBV markers in the evolution of HBsAg-positive liver cirrhosis to primary hepato-cellular carcinoma (PHC) was studied by following up 70 cirrhotic patients from February 1978 to February 1981. Eight out of 19 (42.1%) patients with HBsAg positive liver cirrhosis developed PHC. On the other hand, only 7.8% of the patients with HBsAg negative liver cirrhosis developed PHC. In the HBsAg-positive group only the patients who had already seroconverted to anti-HBe (7 out of 11) or were negative for both HBeAg and anti-HBe (1 out of 3) at the time of first observation and showed a histological picture of inactive cirrhosis, developed PHC during the follow-up period. No HBeAg-positive cirrhosis showed such an evolution. The absence of HBeAg in the sera of patients who developed PHC suggests that active HBV replication becomes increasingly defective during the course of malignant transformation. PMID- 6299917 TI - Effect of dietary fiber, glucomannan, on absorption of sulfonylurea in man. AB - In order to clarify whether a dietary fiber has any effect upon the intestinal absorption of sulfonylurea, changes in plasma concentration of glibenclamide were determined during a six-hour period in nine healthy volunteers who took 2.5 mg of glibenclamide together with a breakfast and 3.9 g of glucomannan in a form of konjac powder and were compared with those of the control experiment in which the same amount of the hypoglycemic agent was given without the dietary fiber. In the control, mean plasma glibenclamide level increased rapidly, reaching a peak at 60 min and decreased gradually thereafter, whereas an increase in plasma glibenclamide level was blunted in the test experiment, thus plasma concentration of glibenclamide being lower at 30, 60, 90 and 150 min compared with the corresponding value of the control (31.7 +/- 24.5 ng/ml vs 76.4 +/- 25.0 ng/ml at 30 min; 51.3 +/- 35.5 ng/ml vs 120.9 +/- 56.0 ng/ml at 60 min; 60.0 +/- 38.8 ng/ml vs 117.4 +/- 53.1 ng/ml at 90 min; 54.0 +/- 31.5 ng/ml vs 100.7 +/- 46.5 ng/ml at 150 min). Mean plasma glucose concentration was significantly lower at 30 min in the test experiment than in the control despite the lower level of plasma glibenclamide in the former. The results suggest that glucomannan may influence the intestinal absorption of glibenclamide. A dietary fiber must be prescribed in due consideration of these facts. PMID- 6299919 TI - Clonidine and GH secretion in insulin dependent diabetes (IDD). PMID- 6299918 TI - Influence of neuroactive drugs on corticosteroid feedback regulation of ACTH secretion in man. AB - Corticosteroid feedback effects on ACTH secretion in man can be manipulated by neuroactive drugs. In patients without endogenous corticosteroids (primary adrenocortical insufficiency) differential and integral feedback effects can be differentiated. When in these patients brain norepinephrine receptor activity was increased by desipramine, the normally negative differential feedback mechanism was converted into a positive one (paradoxical ACTH response). The pre-existing paradoxical ACTH response of patients with Cushing's disease after adrenalectomy was abolished after depletion of norepinephrine granules by means of reserpine. PMID- 6299920 TI - Hepatic sinusoidal cells: new insights and controversies. PMID- 6299921 TI - Serodiagnosis of acute viral hepatitis. PMID- 6299922 TI - Argyrophilic cells in mammary carcinoma. AB - Breast tumor tissues were treated by the Grimelius procedure and examined for the presence of argyrophilic cells. Carcinomas found to contain argyrophilic cells did not include classic carcinoid tumors; the group was, in fact, heterogeneous, comprising poorly differentiated ductal carcinomas, lobular carcinomas, carcinomas of uncertain origin, and colloid carcinomas. Colloid tumors were the most frequently encountered. The prominence of argyrophilic cells in colloid carcinomas raises the possibility that development into mucin-producing cells is propitious for endocrine differentiation. PMID- 6299923 TI - Blood transfusion for the neonatal patient. PMID- 6299924 TI - Monoclonal and polyclonal antibodies against neural antigens: diagnostic applications for studies of central and peripheral nervous system tumors. PMID- 6299925 TI - Numbers of asbestos bodies on iron-stained tissue sections in relation to asbestos body counts in lung tissue digests. AB - Utilization of tissue digestion techniques has demonstrated the presence of large numbers of asbestos bodies within lungs of persons after occupational exposure to asbestos, and smaller numbers in the vast majority of persons with no identifiable exposure. Because of the wide variability in results of such studies among different observers, the presence of more than one asbestos body on light microscopy has been recommended recently as one of the morphologic requirements (together with peribronchiolar fibrosis) for the tissue diagnosis of asbestosis. However, data that correlate the occurrence of asbestos bodies in paraffin embedded tissue sections with the quantification of asbestos bodies by tissue digestion techniques have not been available. The authors counted the asbestos bodies in multiple paraffin-embedded sections of lung tissues stained for iron, and compared those numbers with the asbestos body counts determined by hypochlorite digestion of wet formalin-fixed lung tissue in six cases of asbestosis or asbestos-associated neoplasia. When adjustments were made for asbestos body orientation in tissue sections, shrinkage of sections during processing, and conversion of lung volume to wet weight, the agreement between the two techniques was excellent (r = 0.98, P less than 0.001). An average of two asbestos bodies on 2 X 2 cm (4 cm2) iron-stained tissue sections 5 microns thick is equivalent to approximately 200 asbestos bodies per gram of wet fixed lung tissue. PMID- 6299927 TI - Human and rodent transformed cells are more sensitive to in vitro induction of SCE by N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) than normal cells. AB - The sensitivity to sister chromatid exchange (SCE) induction by N-methyl-nitro-N' nitrosoguanidine (MNNG) in human, mouse, Chinese hamster, and Syrian hamster normal cell strains and in permanent transformed cell lines of the same species was compared. Exponentially growing or growth-inhibited cultures of permanent cell lines transformed spontaneously or by chemical carcinogen or oncogenic virus responded with a higher SCE frequency after MNNG treatment than did normal diploid cell strains. Compared with the normals, exponentially growing Simian virus 40 transformed human fibroblast GM637 had the highest SCE frequency, followed by mouse cell line alpha L929 and Chinese hamster V79-4; the least sensitive were two Syrian hamster cell lines, OBP, derived from transformation of embryo cells with benzo(a)pyrene, and BHK, a spontaneously transformed baby hamster kidney line. Similar results were obtained with cultures arrested in G1 with glutamine-arginine deficient medium. The SCE response observed with transformed cells to carcinogen probably reflects cellular changes associated with the transformed state, such as shortening of the cell cycle, excision repair deficiency, or an increase in DNA replicon size. The current results demonstrating a difference in SCE induction between normal and malignant cells are important since normal or transformed cultured cells are utilized to assess potentially deleterious environmental agents, particularly carcinogens. In general, SCE induction by a specific direct acting carcinogen may be a useful approach for identifying transformed cells with the ability to produce tumors. PMID- 6299928 TI - Loss of high frequency of sister chromatid exchanges in Epstein-Barr virus established lymphoblastoid cell lines from two patients with Bloom's syndrome. AB - Seven lymphoblastoid cell lines were established through transformation by Epstein-Barr virus of peripheral blood lymphocytes from two patients with Bloom's syndrome (BS), the parents of a patient, and normal controls. High baseline levels of sister chromatid exchanges (SCEs) in peripheral blood lymphocytes of BS were reduced to about 10% of their initial value in BS lymphoblastoid cell lines, and the elevation of SCE frequencies induced by ethylmethanesulfonate was the same as in controls. PMID- 6299926 TI - Virus-induced gene mutations of eukaryotic cells. AB - Most animal viruses studied so far induce chromosomal aberrations. In addition, adenoviruses, papovaviruses, and retroviruses are known to induce gene mutations like mutagenic bacteriophages. At least in one case studied retrovirus induced mutagenesis involves gene and/or scripton splitting analogous to the mutagenic mechanism of action of mutatorphage Mu and other movable DNA elements. On the contrary, several results obtained by independent means indicate that Simian virus 40, a papovavirus, does not act by splitting the affected gene but presumably by generation of base pair substitutions or of other minor DNA damages leading to amino acid substitutions. The mechanisms involved are still unknown. There a some hints, however, that these mechanisms might have some step(s) in common with processes leading to malignancy. In fact those viruses proved unequivocally so far to be capable of inducing gene mutations are oncogenic viruses. PMID- 6299930 TI - Acetylphosphatase of Vigna unguiculata subsp. cylindrica. PMID- 6299929 TI - Ovarian substance induces steroid production in cultured granulosa cells. AB - The rat ovary produces an apparently low molecular weight substance that mimics the action of follitropin (FSH) on ovarian granulosa cells in culture. Similar to FSH action, the ovarian substance (OS) induces temporal cell rounding and, later on, intensive progestin production. However, unlike FSH, OS does not induce accumulation of cyclic AMP (cAMP) in the granulosa cells. The ovarian factor cannot be cAMP as its action is not abolished by phosphodiesterase (PDE) treatment. Neither is it a possible PDE inhibitor, as it does not augment cAMP accumulation in granulosa cells or Friend erythroleukemic cells induced by FSH or PGE1, respectively. The factor is still active after heating for 20 min at 90 degrees C but is rapidly inactivated by alkali treatment. In addition, treatment with various proteases did not abolish the steriodogenic activity. These findings suggest a possible novel intraovarian regulator of the granulosa cell function. PMID- 6299931 TI - Modulation of ATPase activity of avian myeloblastosis virus with Ca2+, ATP & its reaction products. PMID- 6299932 TI - Evidence for the inability of the catfish Heteropneustes fossilis & Clarias batrachus, to biosynthesize linoleic & linolenic acids. PMID- 6299933 TI - Hepatitis B surface antigen and cirrhosis in hepatocellular carcinoma. PMID- 6299934 TI - Differential ultrastructure and electron histochemistry of human leukaemic cells, II. Acute and chronic lymphoid leukaemias. PMID- 6299935 TI - The stimulation of superoxide anion production in guinea-pig peritoneal macrophages and neutrophils by phorbol myristate acetate, opsonized zymosan and IgG2-containing soluble immune complexes. AB - The kinetics of superoxide anion production in guinea-pig peritoneal macrophages and neutrophils were determined following in vitro stimulation with phorbol myristate acetate (PMA), opsonized zymosan (OZ) and soluble immune complexes of guinea-pig IgG2 (SIC). Superoxide production was recorded as chemiluminescence (CL) arising from the reductive cleavage of lucigenin. With PMA, both macrophages and neutrophils displayed a two-phase response consisting of a rapid initial burst of CL, which preceded ligand ingestion, followed by a plateau in the CL response which persisted for more than 30 min. By contrast, OZ induced a slow progressive increase in CL in both phagocytes which was consistent with the development of an oxidative burst concomitant with ingestion. The phagocytes differed in their responses to SIC, the macrophages displaying CL kinetics similar to those observed with PMA, whereas the neutrophils responded in the manner observed with OZ. The relationship between disparity in the patterns of macrophage and neutrophil CL responses to SIC and differences in their expression of Fc receptors for IgG2 (Coupland & Leslie, 1983) is discussed. PMID- 6299936 TI - Cytoplasmic requirements for the radio-induced modulation of IgG receptors on B cells. AB - The effect of X-ray irradiation on IgG membrane receptors of B murine lymphocytes was studied. Cells were obtained from peripheral lymph nodes of RK mice and teased in Hank's solution. The cells were irradiated or kept as control samples, incubated at 37 degrees C, with or without drugs with known biochemical action at metabolic or structural levels, and labelled with fluorescein-conjugated anti-IgG antisera. The results show that X-ray irradiation results in a modulation of IgG receptor molecules on B-cells. The disappearance phase which takes only 10 min, is temperature dependent, and is prevented with metabolic inhibitors, microtubular disruptors, db-cAMP and local anesthetics. The re-appearance phase is also temperature dependent but apparently does not have either energy or cytoskeleton participation. The phenomenon is interpreted as partial and transient internalization of IgG molecules in the membrane. PMID- 6299937 TI - Time course of production of low molecular weight viral-neutralizing substance(s) in chickens. PMID- 6299938 TI - Isolation of Enterovirus 70 during conjunctivitis epidemic in Delhi in 1981. PMID- 6299939 TI - Serologic survey for neutralizing antibodies against group B coxsackieviruses in normal population in Delhi area. PMID- 6299940 TI - Clinical spectrum of peripheral neuropathies. PMID- 6299941 TI - Spontaneous rupture of hepatoma. Report of six cases with complete autopsy findings. PMID- 6299942 TI - Correlation of virulence and collagenolytic activity in Entamoeba histolytica. AB - The levels of collagenolytic activity of strains HM-1:1 MSS (HM-1), (HM-1), 200 NIH, and HK-9 of Entamoeba histolytica were compared. Collagen degradation was evaluated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Conditioned media as well as extracts of the highly virulent strain HM-1 effectively degraded native type I collagen. Significantly lower activity was found in the analogous fractions of strains 200-NIH and HK-9, which are not as virulent. The collagenolytic activity of strain HM-1 was associated with the isolated plasma membrane fraction and could be eluted from the membranes by buffers of high ionic strength, indicating that it is not an integral membrane protein. Unlike the vertebrate and the clostridial collagenases, the collagenolytic activity of E. histolytica HM-1 was enhanced in presence of dithiothreitol and was inhibited by N-ethylmaleimide. The correlation between virulence of the individual strains and their collagenolytic activity suggests that collagenase might play a role in pathogenesis of amoebiasis. The localization of the enzyme on the plasma membrane and its presence in the extracellular medium favor this view. PMID- 6299943 TI - Enhanced superoxide anion release from phagocytes by muramyl dipeptide or lipopolysaccharide. AB - Muramyl dipeptide (MDP) and lipopolysaccharide (LPS) from Escherichia coli were tested for the ability to influence superoxide anion (O2-) release from guinea pig phagocytes. Both MDP and LPS alone did not, by themselves, stimulate O2- release by macrophages and polymorphonuclear leukocytes. However, the preincubation of macrophages with MDP or LPS primed the macrophages to release an enhanced amount of O2- when stimulated by cytochalasin E and wheat germ agglutinin. When polymorphonuclear leukocytes were treated in the same way, only LPS showed an enhancing effect. MDP enhanced NADPH oxidase activity of macrophages, which is probably the reason for enhanced O2- release by MDP. PMID- 6299944 TI - Analysis of human rotavirus mixed electropherotypes. AB - Mixed human rotavirus electropherotypes were detected in stool samples from patients with acute gastroenteritis in Santiago, Chile. These electropherotypes accounted for 10% of 149 samples studied. The finding of extra RNA fragments with respect to the regular 11 genome segments suggests the possibility of simultaneous or sequential infection by more than one electropherotype in a single diarrhea event or occurrence of modification in the length of the RNA segments during an infection. These possibilities arose from gel electrophoretic analysis of unique and sequential samples of human rotavirus genome RNA. PMID- 6299945 TI - Tumor-dependent resistance of rat peritoneal macrophages to herpes simplex virus. AB - By their position at sites of initial infection and their wide distribution in major organs of the body, macrophages may be decisive in determining the susceptibility or resistance of the host to virus infection. Macrophage restriction of virus replication has been shown to be closely related to virus strains or virus types and to the age of the infected host. We report the effects of the development of a solid tumor in rats on intrinsic in vitro macrophage activity against herpes simplex virus type 1. The results obtained with the infectious center assays and the analysis of single-cycle growth curves of herpes simplex virus type 1 in macrophages obtained from normal and tumor-bearing rats showed a depression of antiviral activity of macrophages from tumor-bearing rats. The possibility of immunomodulation by bacterial adjuvants on tumor-bearing rats and the effects on the antiviral activity of peritoneal macrophages were furthermore demonstrated. PMID- 6299946 TI - Structure and biological properties of solubilized envelope proteins of Bordetella pertussis. AB - The structure and biological properties of solubilized envelope proteins of Bordetella pertussis have been examined. Several envelope proteins were found to be specific for phase I strains of B. pertussis and could be isolated by selective detergent extraction. These proteins had molecular weights of 90,000, 86,000, 81,000, 33,000, 31,000, and 30,000 and were reduced or absent in envelope preparations from Bordetella bronchiseptica, Bordetella parapertussis, or phase IV strains of B. pertussis. When the envelope preparations from phase I B. pertussis were assayed in the mouse intracerebral protection test they were found to be highly protective, and there was a strong correlation between the protective potency and the lymphocytosis-promoting factor (LPF) content of different preparations. Treatment with glutaraldehyde reduced the LPF activity, toxicity, and protective potency of the envelope extracts. Similarly affinity chromatography of envelope proteins on columns of haptoglobin coupled to Sepharose 4B reduced both the LPF content and the protective potency. The addition of a small amount of purified LPF to the haptoglobin-treated proteins restored the protective potency. The LPF by itself was nonprotective, indicating a potentiating role of LPF in the mouse intracerebral challenge test. PMID- 6299947 TI - Role of myeloperoxidase in luminol-dependent chemiluminescence of polymorphonuclear leukocytes. AB - When polymorphonuclear leukocytes (PMNL) and soluble or particulate matter interact, the cells produce chemiluminescence, linked to activation of the oxidative metabolism of the cells. PMNL isolated from a patient with a myeloperoxidase deficiency were found to produce almost no luminol-dependent chemiluminescence, despite a pronounced production of superoxide anions (O2-). The chemotactic peptide formylmethionyl-leucyl-phenylalanine induced a two-peak chemiluminescence response in control PMNL. The response was modified, both in magnitude and in time-course, when the cells were incubated at 22 degrees C for 120 min. Addition of purified myeloperoxidase to the PMNL lacking this enzyme, before stimulus addition, resulted in a chemiluminescence response. In the response to formylmethionyl-leucyl-phenylalanine, only one peak, corresponding to the initial peak of control PMNL, was found. This indicated that luminol dependent chemiluminescence is dependent on and directly related to the presence of myeloperoxidase in PMNL and that both intra- and extracellularly located myeloperoxidase has to be taken into account when interpreting the cellular response assayed as chemiluminescence. PMID- 6299948 TI - Characteristics of mononuclear phagocytes mediating antilisterial resistance in splenectomized mice. AB - The characteristics of mononuclear phagocytes mediating resistance to infection with Listeria monocytogenes during the early phase (up to 48 h) of the response were investigated in mice of the A strain that had undergone splenectomy. Although irradiation in the sham-operated host had no effect on its antilisterial response when administered immediately before infection, it markedly reduced the ability of the splenectomized host to resist listerial challenge. This effect of radiation was demonstrable in the high-dose range (600 r) and could not be reversed immediately by repopulation with 20 x 10(6) syngeneic nucleated bone marrow cells. Administration of silica 24 h before infection profoundly enhanced the growth of L. monocytogenes in the liver of splenectomized mice. Shielding of the liver, but not the bone marrow, protected the splenectomized host against the effects of radiation, indicating that the cell population responsible for mediating the enhanced antilisterial resistance resides in the liver. The enhanced antilisterial resistance of splenectomized mice was specifically because of the absence of the spleen and not merely because of the removal of a favorable replicating environment for listeria organisms. PMID- 6299949 TI - Frequency of cytotoxic T lymphocyte precursors to herpes simplex virus type 1 as determined by limiting dilution analysis. AB - The conditions for establishing a limiting dilution assay to measure cytotoxic T lymphocyte precursors (CTL-P) against herpes simplex virus type 1 (HSV-1) were determined. Analysis by Poisson statistics demonstrated that the estimated frequency of HSV-1-reactive cells in the spleens of normal mice was less than 1/250,000. In contrast, mice immunized previously with infectious HSV-1 demonstrated a CTL-P frequency between 1/3,500 and 1/15,670. The generation of a maximum cytotoxic T lymphocyte response required that mice be primed in vivo with infectious virus. Immunization with inactivated virus either failed to elicit detectable CTL-P frequencies or gave frequencies markedly less than those induced with infectious virus. To obtain positive cultures, the responder cell population had to be exposed to stimulator splenocytes expressing viral antigens. Normal splenocytes without virus or normal splenocytes with T cell growth factor did not result in significant cytotoxicity. Split culture analysis comparing cytotoxicity against syngeneic and allogeneic virus-infected targets provided evidence for specificity, H-2 restriction, and the T cell nature of the CTL-P. It was determined that precursors were eliminated by treatment with anti-Thy 1, Lyt 2.1, or Lyt 1.1, indicating the CTL-P were Lyt 1(+)2(+) cells. Cytotoxicity was reduced after treatment of the responders with anti-Lyt 2 plus complement, which gave further evidence of the T cell nature of the cytotoxic T lymphocytes. These experiments demonstrated the feasibility of using the limiting dilution approach as a highly sensitive and quantitative means to measure the cell-mediated immune response to HSV-1 antigens.